PMID- 12074732 TI - Cutaneous gene transfer for skin and systemic diseases. AB - Recent progress in molecular genetics has illuminated the basis for a wide variety of inherited and acquired diseases. Gene therapy offers an attractive therapeutic approach capitalizing upon these new mechanistic insights. The skin is a uniquely attractive tissue site for development of new genetic therapeutic approaches both for its accessibility as well as for the large number of diseases that are amenable in principle to cutaneous gene transfer. Amongst these opportunities are primary monogenic skin diseases, chronic wounds and systemic disorders characterized by low or absent levels of circulating polypeptides. For cutaneous gene therapy to be effective, however, significant progress is required in a number of domains. Recent advances in vector design, administration, immune modulation, and regulation of gene expression have brought the field much nearer to clinical utility. PMID- 12074733 TI - Perioperative myocardial infarction in noncardiac surgery: the diagnostic and prognostic role of cardiac troponins. AB - Despite the number of technologies used, the diagnosis of perioperative myocardial infarction is still a challenge. Studies conducted in surgical series have demonstrated that cardiac troponins (cTns) have both a superior diagnostic sensitivity and specificity, compared with other traditional techniques, and an independent power to predict short- and long-term prognosis. Nevertheless, some points need to be clarified. They include the usefulness of cTns in patients with end-stage renal failure; the standardization of the cTns cut-off for the diagnosis of myocardial injury; the timing of postoperative blood samplings; the cost-effectiveness of a screening in asymptomatic patients; and the possible therapeutic strategies. PMID- 12074734 TI - Development of hypertension over 6 years in a birth cohort of young middle-aged men: the Cardiovascular Risk Factor Study in southern Sweden (CRISS). AB - OBJECTIVES: To explore the development of hypertension (HT) in a cohort of young middle-aged men. DESIGN: Prospective birth-cohort study of men surveyed over 6 years. SETTING: Helsingborg County Hospital, Sweden, 1990-97. SUBJECTS: A total of 628 men born in 1953-54, all surveyed at 37, 40 and 43 years of age. MAIN OUTCOME MEASURES: Systolic blood pressure (SBP), diastolic blood pressure (DBP), S-cholesterol, body mass index (BMI), alcohol consumption, ethnicity. HT was defined as SBP > or = 140 mmHg and/or DBP > or = 90 mmHg, or ongoing treatment. Using SBP < 130 mmHg and DBP < 85 mmHg as reference, the odds of conversion to HT in men with high normal blood pressure (BP) (SBP 130-139 mmHg and DBP 85-89 mmHg) was investigated. RESULTS: At age 37, 243 men (39%) had reference BP, 167 (26%) had high normal BP and 218 (35%) were hypertensive. Corresponding numbers at age 40 were 265 (42%), 166 (27%) and 197 (31%); and at age 43, 180 (29%), 142 (22%) and 306 (49%), respectively. High normal BP at baseline was associated with the development of HT both at age 40 (odds ratio (OR)=2.45 confidence interval (CI): 1.42-4.22) and at age 43 (OR=2.46, CI: 1.59-3.80), independent of other cardiovascular disease risk factors and ethnicity. The progression to HT was predicted also by S-cholesterol, alcohol consumption, BMI and weight gain. CONCLUSIONS: Over a short-term period, a substantial proportion of young middle aged men with high normal BP develop HT with overweight and alcohol consumption as important determinants. These findings have implications for the prevention, screening and medical care of HT in this target population. PMID- 12074735 TI - Is lipoprotein(a) a predictor for survival in patients with established coronary artery disease? Results from a prospective patient cohort study in northern Sweden. AB - OBJECTIVES: Lipoprotein(a) [Lp(a)] is a known risk factor for the development of atherosclerosis. The aim of the present study was to test the importance of Lp(a) as a predictor for the further prognosis in patients with established coronary artery disease. DESIGN: A prospective patient cohort study was carried out. SETTING AND SUBJECTS: The cohort consists of 1216 patients who were examined with coronary angiography at the University Hospital in Umea, Sweden, because of stable effort angina. MAIN OUTCOME MEASURES: Lipids, Lp(a), fibrinogen, antithrombin III (AT III), sedimentation rate and clinical data were registered at angiography. After a mean follow-up time of 6.7 years information on survival was collected from the municipal census lists and death certificates were examined. Total mortality and mortality because of cardiovascular disease were both used as outcome variables in the survival analyses. RESULTS. The total mortality in the patient cohort was 16.4%. An Lp(a) level of 300 mg L-1 or more was found in 30% of the study population and was found to be an independent predictor for death. A high fibrinogen, a low AT III level, a depressed left ventricular function and a high coronary obstruction score were other significant independent predictors of death. Total cholesterol, HDL- and LDL-cholesterol were not related to survival in this study, but a substantial proportion of the population probably received lipid-lowering agents during the study period. CONCLUSIONS: An Lp(a) level exceeding 300 mg L-1 indicates a poor further prognosis and may help to identify patients who probably need powerful secondary prevention programmes to improve their prognosis. PMID- 12074736 TI - Validity of D-dimer tests in the diagnosis of deep vein thrombosis: a prospective comparative study of three quantitative assays. AB - OBJECTIVES: To assess the diagnostic reliability of a new quantitative D-dimer assay (VIDAS New) and an established quick test (Nycocard D-dimer assay) in the diagnosis of deep vein thrombosis (DVT) compared with ultrasonography. A third assay (Auto Dimer) became available during sample collection and has been included in the final assessment. The diagnostic performance of the Auto Dimer assay was evaluated on three different coagulation analysers. DESIGN: A clinical prospective study of patients admitted to hospital for evaluation of DVT. Setting. The admission ward at Aalborg Hospital. Subjects. A total of 113 outpatients with suspected DVT. Main outcome measures. Compression ultrasonography was used as the reference method for a diagnosis of DVT and compared with different D-dimer assays. The results were expressed as sensitivity, specificity, positive predictive value and negative predictive value (NPV). RESULTS: Deep vein thrombosis was established in 49 patients (43%). Two D dimer assays (VIDAS New and Auto Dimer) showed sensitivities of 90 and 88%, specificities of 42 and 44%, and NPV's of 85 and 83%, respectively. The Nycocard D-dimer assay showed a sensitivity of 63%, specificity of 67% and NPV of 71%. CONCLUSIONS: The diagnostic performance of VIDAS New and the Auto Dimer D-dimer assays is almost identical, but this study suggests that neither of the D-dimer assays is suitable as the only screening method for DVT, in a situation with a high pretest probability of DVT. This call for a differential strategy that distinguishes between cases of low and high clinical probability using either a D dimer test or ultrasonography. Abbreviations DVT, deep venous thrombosis, NPV, negative predictive value, PPV, positive predictive value PMID- 12074737 TI - Dyspeptic symptoms in middle-aged to old adults: the role of Helicobacter pylori infection, and various demographic and lifestyle factors. AB - OBJECTIVES: To evaluate the role of factors supposed to interfere with the gastric mucosa like Helicobacter pylori, smoking and analgesics in symptom generation in a population-based sample of middle-aged to old. SUBJECTS: A total of 288 subjects aged 50-85 (mean 65 +/- 7.2) attending a General Science orientated Continuing Education Programme at the University of Ulm. METHODS: Helicobacter pylori infection was determined by the 13C-urea breath test. Demographic data, gastrointestinal symptoms and factors supposed to be involved in symptom generation were collected by a standardized questionnaire. The association between various demographic and lifestyle factors with a gastrointestinal symptom score was quantified by odds ratios (ORs) using multiple logistic regression. RESULTS: Use of nonsteroidal anti-inflammatory drugs (OR=1.2; 95% CI=0.7-2.1) was no risk factor for abdominal symptoms, whereas female sex (OR= 1.6; 95% CI=0.9-2.9) and current smoking (OR= 3.7; 95% CI=0.9 15.4) were associated with a high symptom score. Age 70 years and over was significantly associated with a lower symptom score (OR=0.4; 95% CI=0.2-0.9). However, H. pylori infection was a risk factor for a high symptom score in that age group (OR=4.3; 95% CI=1.2-14.8), whereas no such association (and even a tendency to reduced symptom levels in infected subjects) was observed in younger age groups. CONCLUSION: The role of H. pylori infection in symptoms may vary by age. The loss of symptoms in the elderly may delay detection of gastroduodenal pathology and appropriate treatment in older adults. PMID- 12074738 TI - Exacerbated chronic obstructive pulmonary disease: a frequently unrecognized condition. AB - OBJECTIVES: To assess to which extent exacerbated chronic obstructive pulmonary disease (COPD) remains unrecognized in the emergency department, which factors account for misdiagnosis and which are the effects of misdiagnosis on patient's management. DESIGN: Retrospective study and stratified random sampling method as selection criterion. SETTING: University Hospital. SUBJECTS: Eighty patients representative of those discharged from the wards of medicine with a diagnosis of exacerbated COPD and 72 having a discharge diagnosis of coronary artery disease (CAD) were studied. MAIN OUTCOME MEASURES: Degree of concordance between admission and discharge diagnosis; presenting symptoms and signs of patients correctly or incorrectly classified on admission; impact of diagnostic procedures carried out by the physician on call on patient's management. RESULTS: The correct diagnosis was missed on admission in 13/80 COPD and 3/72 CAD patients (chi(2): 5.87, P=0.015). The prevalence of the following presenting features distinguished the 67 COPD patients who were correctly classified on admission from the remaining 13: severe weakness (21 vs. 10, chi(2): 9.53, P=0.002), dyspnea (60 vs. 3, chi(2): 28.75, P < 0.001), and limb oedema (14 vs. 6, chi(2): 3.70, P=0.054). Critical hypoxemia was diagnosed and treated only after admission in 19 COPD patients. CONCLUSIONS: Exacerbated COPD frequently escapes recognition in the emergency room, mainly if severe weakness and limb oedema are its presenting features. Arterial blood gas analysis is not systematically performed in the emergency room and, consequently, oxygen therapy is either not administered or given to selected COPD patients on an empirical basis. PMID- 12074739 TI - Muscle strength, Na,K-pumps, magnesium and potassium in patients with alcoholic liver cirrhosis -- relation to spironolactone. AB - OBJECTIVES: To evaluate the muscle strength in relation to muscle contents of magnesium (Mg), potassium (K) and sodium, potassium (Na,K)-pumps in patients with alcoholic cirrhosis. DESIGN: An open cross-sectional study. SETTING AND SUBJECTS: Fifty-one consecutive patients with liver cirrhosis admitted to the Department of Hepatology, Aarhus University Hospital, Denmark, and 28 age- and sex-matched healthy control subjects. MAIN OUTCOME MEASURES: Biopsies of skeletal muscle were performed in patients and controls for measurements of Mg, K, and Na,K-pumps. Furthermore, maximum isokinetic knee extension and skeletal muscle mass were evaluated. RESULTS: Muscle mass, muscle strength, muscle Mg and muscle K were substantially reduced in the patients (P < 0.01, all), and fell with increasing severity of the liver disease reflected in the Child-Pugh (C-P) class. Patients treated with spironolactone for 2 weeks or more, had increased muscle strength, muscle Mg and content of Na,K-pumps, compared with the rest of the patients (P < 0.05, all). In a multivariate analysis of the patients, skeletal muscle mass, muscle Mg and daily alcohol consumption (g) were independent predictors of isokinetic muscle strength (P < 0.05, all). CONCLUSIONS: Patients with alcoholic liver cirrhosis showed considerably reduced muscle strength and muscle Mg was an independent predictor of muscle strength. Surprisingly, in the spironolactone treated patients, muscle weakness was less pronounced, possibly because of the action of spironolactone on muscle Mg, K and Na,K-pump content. PMID- 12074740 TI - Atrial fibrillation or flutter and stroke: a Danish population-based study of the effectiveness of oral anticoagulation in clinical practice. AB - OBJECTIVES: A pooled analysis of randomized trials has shown that oral anticoagulation therapy reduces the risk of ischaemic stroke with 68% in patients with atrial fibrillation. We examined the effectiveness of oral anticoagulation on risk of stroke of any nature (fatal and nonfatal ischaemic and/or haemorrhagic stroke) in patients with nonvalvular atrial fibrillation or flutter living in the County of North Jutland, Denmark. DESIGN: Cohort study. SUBJECTS AND METHODS: We used the Hospital Discharge Registry covering the county (490 000 inhabitants) from 1991 to 1998 to identify 2699 men and 2425 women with atrial fibrillation or flutter, aged 60-89 years. Data on prescriptions of anticoagulation were obtained from the National Health Service. We defined use of oral anticoagulation as date of prescription or reiteration plus 90 days. Patients were followed in the County Hospital Discharge Registry until a diagnosis of stroke (fatal and nonfatal ischaemic and/or haemorrhagic stroke), emigration, death or the end of 1998. We used Cox regression analyses to estimate the relative risk of stroke associated with use of oral anticoagulation compared with no use, adjusted for age, diabetes and underlying cardiovascular diseases. RESULTS: Eight hundred and thirty-eight of 2699 men (31%) and 552 of 2425 women (23%) with atrial fibrillation had one or more recorded prescriptions of oral anticoagulation. The incidence rates of stroke were 31 per 1000 person-years of follow-up in men, and 30 per 1000 person years of follow-up in women. The adjusted relative risks of stroke during anticoagulation were 0.6 [95% confidence interval (CI) 0.4-1.0] in men, and 1.0 (95% CI 0.7-1.6) in women compared with nonuse periods. CONCLUSIONS: The effectiveness of oral anticoagulation in clinical practice may be lesser than the efficacy of oral anticoagulation reported from randomized trials. PMID- 12074741 TI - The enigma of increased non-cancer mortality after weight loss in healthy men who are overweight or obese. AB - OBJECTIVE: To study effects on non-cancer mortality of observational weight loss in middle-aged men stratified for body mass index (BMI), taking a wide range of possible confounders into account. DESIGN: Prospective, population based study. SETTING: Male population of Malmo, Sweden. PARTICIPANTS: In all 5722 men were screened twice with a mean time interval of 6 years in Malmo, southern Sweden. They were classified according to BMI category at baseline (<21, 22-25, overweight: 26-30, and obesity: 30+ kg m(-2)) and weight change category until second screening (weight stable men defined as having a baseline BMI +/- 0.1 kg m(-2) year-1 at follow-up re-screening). MAIN OUTCOME MEASURES: Non-cancer mortality calculated from national registers during 16 years of follow-up after the second screening. Data from the first year of follow-up were excluded to avoid bias by mortality caused by subclinical disease at re-screening. RESULTS: The relative risk (RR; 95% CI) for non-cancer mortality during follow-up was higher in men with decreasing BMI in all subgroups: RR 2.64 (1.46-4.71, baseline BMI <21 kg m(-2)), 1.39 (0.98-1.95, baseline BMI 22-25 kg m(-2)), and 1.71 (1.18 2.47, baseline BMI 26+ kg m(-2)), using BMI-stable men as reference group. Correspondingly, the non-cancer mortality was also higher in men with increasing BMI, but only in the obese group (baseline BMI 26+ kg m(-2)) with RR 1.86 (1.31 2.65). In a subanalysis, nonsmoking obese (30+ kg m(-2)) men with decreased BMI had an increased non-cancer mortality compared with BMI-stable obese men (Fischer's test: P=0.001). The mortality risk for nonsmoking overweight men who increased their BMI compared with BMI-stable men was also significant (P=0.006), but not in corresponding obese men (P=0.094). CONCLUSIONS. Weight loss in self reported healthy but overweight middle-aged men, without serious disease, is associated with an increased non-cancer mortality, which seems even more pronounced in obese, nonsmoking men, as compared with corresponding but weight stable men. The explanation for these observational findings is still enigmatic but could hypothetically be because of premature ageing effects causing so-called weight loss of involution. PMID- 12074742 TI - Kikuchi-Fujimoto's disease: report of familial occurrence in two human leucocyte antigen-identical non-twin sisters. AB - Kikuchi-Fujimoto's disease (KFD) is a self-limiting condition usually presenting as cervical lymphadenopathy, with fever and leukopenia, in young adult females. No specific aetiology has been identified yet, although some authors believe that KFD is a 'forme fruste' of systemic lupus erythematosus. Certain human leucocyte antigen (HLA) types have also been associated with KFD. We herein report KFD in two non-twin sisters with HLA-identical phenotype, who presented 10 years apart. Neither patient had evidence of recent infection or connective tissue disease. The familial occurrence emphasizes the possibility of genetic predisposition and calls for a more extensive search for a specific cause of KFD. PMID- 12074743 TI - Rescue percutaneous transluminal coronary angioplasty in a patient with a single coronary artery arising from the right Sinus Valsalvae: previously unreported scenario and review of literature. AB - This is the first description of treatment with coronary stenting of a patient with a single coronary artery originating from the right Sinus Valsalvae and suffering from acute inferior myocardial infarction. Angiography showed the following: the right coronary artery (RCA) had a normal course, whilst the left anterior descending (LAD) as well as the left circumflex (LCX) branches both originated separately from the proximal RCA, which served as a common mixed trunk. The LAD crossed to the left in front of the right ventricular outflow tract, whilst the LCX, taking a retroaortic course to the atrioventricular groove, had a distal occlusion. According to an anatomically based classification considering all imaginable variations, this configuration corresponds to a II-D-1 pattern, which previously has been described only in a single postmortem case. After the LCX was reopened with a guide wire, a coronary stent was successfully inserted, resulting in TIMI-3 flow. Recovery was uneventful. PMID- 12074744 TI - Unexpected toxicity induced by magnesium orotate treatment in congenital hypomagnesemia. PMID- 12074745 TI - A critical view of spirituality and spiritual assessment. PMID- 12074747 TI - Nurse practitioner knowledge of complementary alternative health care: foundation for practice. AB - AIM: With the increasing use of complementary alternative treatments by the general public in the United States of America (USA), it is critical that nurse practitioners have the most up-to-date information about the use and safety of these modalities. A strong knowledge base is crucial in delivering competent and culturally sensitive care, yet the level and source of nurse practitioner knowledge in this area is largely unknown. The purpose of this study was to investigate the level and source of nurse practitioner knowledge of complementary alternative health care practices, as well as their referral practices involving these treatments. METHODS: Using a cross-sectional descriptive research design, a sample of 151 nurse practitioners from Missouri and Oregon completed an adapted version of Sapp's self-administered survey that explored these issues. Evaluation of the psychometric properties of the adapted instrument demonstrated good face validity, test-retest reliability (kappa = 0.81) and internal consistency reliability were 0.90 and 0.92 for two subscales with continuous response categories. FINDINGS: Eighty-three percent of the nurse practitioners recommended complementary alternative treatments to their patients with the most frequent being massage therapy, chiropractic care, acupuncture or acupressure, nutritional therapy and herbal treatment. However, only 24% reported that formal nurse practitioner education was a source of knowledge about these treatments. Instead, over 60% relied on their personal experiences for this knowledge, as well as lay and professional journals. CONCLUSION: Nearly 9 out of 10 nurse practitioners recommend the use of complementary alternative therapies to patients, but their source of knowledge is not derived from professional education. Attention needs to be given to increasing content about complementary alternative therapies in formal academic programs, professional conferences and in-service education opportunities. PMID- 12074748 TI - A review of symptoms of coronary artery disease in women. AB - BACKGROUND: Research on heart disease has increasingly included information on women's experiences. A number of recent studies present frequencies and comparisons of symptoms between men and women and there appears to be some variability in the symptoms especially among women. Even with acute myocardial infarction (AMI) where anticipated symptoms are more clear-cut, women can have vague or nonclassic symptoms. AIM AND METHOD: Selected medical and nursing research on cardiac symptoms is examined for information on the cardiac warning symptom experiences unique to women. A search of the literature between 1995 and 2000 was done using CINAHL and MEDLINE. Terms used for the search included: cardiac symptoms, women's symptoms and symptom perception. The findings from this review are used to suggest implications for clinical practice. FINDING: Women experiencing AMI present with a variety of symptoms including chest pain. Less obvious symptoms include; fatigue, shortness of breath, back pain, oedema, and transient non-specific chest discomfort. These less dramatic and non-specific symptoms do not necessarily prompt further assessment for coronary disease in women. CONCLUSION: Cardiac screening of women who present with cardiac risk factors and careful attention to less anticipated symptoms are critical factors that can improve the rapid identification of coronary disease in women. The unique physiological and sociological differences between women and men make further study of women's symptom experiences and perceptions important for health care providers. Further study of gender and ethnic differences in symptom patterns and recognition will help to improve screening and earlier identification of cardiac problems in women patients especially those without chest pain as a prodromal symptom. PMID- 12074749 TI - Critical care cardiovascular nurse expert and novice diagnostic cue utilization. AB - AIM: Critical care cardiovascular (CCCV) nursing diagnostic expertise was the focus of this research. The purpose of the study was to compare diagnostic cue utilization between expert and novice CCCV nurses. BACKGROUND: Knowledge related to objective measures of clinical nursing diagnostic expertise would enhance effective and efficient recognition, utilization, and reward of clinical expertise. METHODS: Five CCCV written simulations served as instruments in the study. Diagnostic content areas included left ventricular dysfunction, cardiac tamponade, sepsis, right ventricular failure, and hypovolemia related to internal abdominal haemorrhage. The sample was composed of 23 expert and 23 novice nurses. After reading each simulation, subjects were asked to verbally recall the simulation, give an impression of the predominant problem or diagnosis, and give a diagnostic explanation. Verbal recalls were audio-taped for protocol analysis. Diagnostic accuracy and cue utilization were determined through comparisons of subjects' recalled diagnoses and cues with results from an expert panel review consensus. The major variable was the mean recalled proportion of highly relevant cues to total cues (HRC/TC) on accurately diagnosed simulations. Chi-square analysis revealed that diagnostic accuracy was greater with experts than with novices. Differences between and among simulations, expertise, accuracy and the mean proportion of highly relevant cues to total cues were examined with a 4 x 2 x 2 factorial analysis of variance. RESULTS: When considering all accurately diagnosed simulations, experts had a higher HRC/TC than novices. The major limitations were the use of low fidelity written simulations and virgin verbal protocol methods. CONCLUSION: The findings generally support the idea that the development of diagnostic expertise is associated with the ability to focus on highly relevant cues. PMID- 12074750 TI - Tracheal suctioning: an exploration of nurses' knowledge and competence in acute and high dependency ward areas. AB - BACKGROUND AND RATIONALE: With an increasing demand for intensive care beds more nurses in acute and high dependency wards will be expected to care competently for patients with tracheostomy tubes. Tracheal suctioning is an essential aspect of effective airway management. However, this has many associated risks and complications, ranging from trauma and hypoxaemia to, in extreme cases, cardiac arrest and death. It is imperative that nurses are aware of these risks and are able to practice according to current research recommendations. Aims. This study was designed to explore nurses' knowledge and competence in performing tracheal suctioning in acute and high dependency ward areas and to investigate discrepancies between knowledge and practice using method triangulation. METHODS: Twenty-eight nurses were observed using nonparticipant observation and a structured observation schedule. Each subject was interviewed and questioned about their tracheal suctioning practices, and subsequently completed a knowledge based questionnaire. Scores were allocated for knowledge and practice. FINDINGS: The findings demonstrated a poor level of knowledge for many subjects. This was also reflected in practice, as suctioning was performed against many of the research recommendations. Many nurses were unaware of recommended practice and a number demonstrated potentially unsafe practice. In addition, there was no significant relationship between knowledge and practice. However, during the interviews, many nurses were able to provide a rationale for specific aspects of practice that were perhaps not based on current research recommendations. CONCLUSIONS: The study raised concern about all aspects of tracheal suctioning and has highlighted the need for changes in practice, clinical guidelines and focused practice-based education. PMID- 12074751 TI - Acute care nurses' perceptions of barriers to using research information in clinical decision-making. AB - AIM: To examine the barriers that nurses feel prevent them from using research in the decisions they make. BACKGROUND: A sizeable research literature focusing on research utilization in nursing has developed over the past 20 years. However, this literature is characterized by a number of weaknesses: self-reported utilization behaviour; poor response rates and small, nonrandom sampling strategies. DESIGN: Cross-case analysis involving anonymised qualitative interviews, observation, documentary audit and Q methodological modelling of shared subjectivities amongst nurses. The case sites were three large acute hospitals in the north of England. One hundred and eight nurses were interviewed, 61 of whom were also observed for a total of 180 h, and 122 nurses were involved in the Q modelling exercise (response rate of 64%). RESULTS: Four perspectives were isolated that encompassed the characteristics associated with barriers to research use. These related to the individual, organization, nature of research information itself and environment. Nurses clustered around four main perspectives on the barriers to research use: (1) Problems in interpreting and using research products, which were seen as too complex, 'academic' and overly statistical; (2) Nurses who felt confident with research-based information perceived a lack of organizational support as a significant block; (3) Many nurses felt that researchers and research products lack clinical credibility and that they fail to offer the desired level of clinical direction; (4) Some nurses lacked the skills and, to a lesser degree, the motivation to use research themselves. These individuals liked research messages passed on to them by a third party and sought to foster others' involvement in research-based practice, rather than becoming directly involved themselves. CONCLUSIONS: Rejection of research knowledge is not a barrier to its application. Rather, the presentation and management of research knowledge in the workplace represent significant challenges for clinicians, policy-makers and the research community. PMID- 12074752 TI - Application of a computerized nursing care plan system in one hospital: experiences of ICU nurses in Taiwan. AB - AIM OF THE STUDY: The purpose of this paper is to present ICU nurses' experiences with a computerized nursing care plan system at a medical centre in Taiwan. BACKGROUND: Computerized nursing care plans have been implemented in recent years, but users' perceptions of this technology have not been comprehensively explored. It is believed that by taking into consideration users' experiences with computer use, strategies and programs can be developed to help users adapt to new systems. METHODS: A qualitative study, involving one-to-one interviews, was conducted with 12 Registered Nurses. Data were analysed according to Miles and Huberman's data reduction, data display and conclusion verification process. FINDINGS: Themes related to the clinical impact of system use were: 'saves paper/time', 'time-consuming for print-outs', 'de-individualization of care plan', 'routine/paperwork requirement', 'no consensus for nursing diagnoses', and 'guidelines for the novice'. CONCLUSIONS: Nurses prefer tools that can help them save time with paperwork and focus on targeting patient problems. With careful assessment and evaluation of nurses' experiences in the use of clinical computer systems, the success of computer implementation can be enhanced. PMID- 12074753 TI - Vertical equity in service provision: a model for the Irish public health nursing service. AB - AIMS: This paper analyses the policy and conceptual basis of public health nursing service provision in the Republic of Ireland and situates it within an international context. It draws on the principles of horizontal and vertical equity in proposing a new model of public health nursing service provision. It gives the reader an understanding of a model of service delivery underpinned by the principle of vertical equity. BACKGROUND ISSUES: The Public Health Nurse in the Republic of Ireland has a wide remit encompassing primary, secondary and tertiary care at the level of the individual, family and community. The changing sociological and demographic nature of society in Ireland has impacted on a service that has largely remained unchanged since 1966. Since 1997 four review bodies have provided recommendations that are incompatible with each other. There remains a need to find a solution to the overwhelming demands placed on the public health nursing service in the Republic of Ireland. KEY ISSUES: The public health nursing service goes beyond the provision of a purely clinical nursing service. Communities differ in demography, epidemiology, environment, history, composition, support and most importantly needs. Using three exemplars a new model for the public health nursing service is explicated. This model has as its main focus the needs of the community it serves. CONCLUSIONS: A service underpinned by the principle of vertical equity can be used to deliver locally based, needs driven public health nursing services. The application of such a model would make the public health nursing service more flexible and responsive to local need. Public health nursing composition and provision must be determined using the principal of vertical equity determined by the needs of the community it serves. PMID- 12074754 TI - Care of women with breast cancer on a surgical ward: nurses' opinions of the need for support for women, relatives and themselves. AB - BACKGROUND: In Sweden women with newly diagnosed breast cancer are admitted to surgical wards in order to undergo surgery and receive postoperative care. On these wards, nursing staff take care of women both with newly diagnosed breast cancer and those with cancer in advanced stages. Nurses have to meet the varying needs of patients and their relatives. AIM: To describe nurses' opinions of the need for care and support for women and their relatives in connection with surgery for breast cancer, as well as their own need for support on a surgical ward. METHODS: Thirty-one nurses from a surgical ward participated in semi structured interviews. The interviews were tape-recorded and transcribed verbatim. Thereafter a step-by-step, qualitative content analysis was carried out. RESULTS: The nurses described the need to talk and receive information as being the most important among women and their relatives, as well as among themselves. Only a few nurses mentioned the need for physical care among the women. Contact with relatives was described as being almost nonexistent. There was a discrepancy between what nurses described as important needs and how these needs were provided for. CONCLUSION: This study shows that what the nurses described as being the most important needs, and the way how these needs were provided for, was more often seen from a theoretical point of view with few examples of self-experienced situations in the daily care. Needs among women and their relatives seemed to be not fully known to nurses and therefore, possibly, were not met. Nurses themselves had a pronounced need for support, which was sometimes unsatisfactorily met. PMID- 12074755 TI - Older Swedish women's experiences of living with symptoms related to Parkinson's disease. AB - BACKGROUND: Approximately 1% of the population over 65 years of age is afflicted with Parkinson's disease (PD). The number of patients with the disease will most probably increase in the future because of the increased longevity of the population. There is no curative therapy for the disease. AIM: To explore women's experiences of living with symptoms related to PD, and to analyse how the symptoms influence their quality of life. METHODS: A phenomenological-hermeneutic method, inspired by the philosophy of Ricoeur was used. The study focuses on eight women between 63 and 80 years of age who have been diagnosed with PD for 5 15 years. The women all lived at home, were moderately to severely disabled and cognitively intact. FINDINGS: In the analysis, four themes emerged: (1) wish for a stable body image; (2) wish to keep traditional female competence; (3) need to feel accepted for the person she is; and (4) perceived stigmatization. All themes showed that trying to adapt to unpredictable fluctuations in physical and psychosocial competence has a great impact on the females' lives. It became clear that fluctuations in competence and not knowing when to expect impaired mobility are connected with frustration and social withdrawal. CONCLUSION: The experience of PD from eight disabled female patients has revealed that the disease affects quality of life not only for the person who has it, but also for her family. Thus, it is equally urgent that patients, relatives, nurses and caregivers gain better knowledge and more understanding of PD, so as to ease the impact of the disease on the patient's daily life. PMID- 12074756 TI - Elisions in the field of caring. AB - BACKGROUND: Contemporary research into caring in nursing was criticized in the pages of this journal by John Paley. He charged that the study of caring has not been advanced by research which, he reckoned, merely generates endless lists of terms to describe caring. He also argued that research in the field was largely flawed by confusion over the difference between things said about caring and the act of caring itself. THE PRESENT PAPER: We have analysed Paley's criticism. Essentially, he is criticizing the whole field of survey research. The scientific process is underpinned by the implicit understanding that any field moves forward cautiously. In the social sciences multiple perspectives enrich understanding of phenomena and often confirm previous perceptions. The lack of any alternative approach from Paley is evident. Examples from psychology, where seemingly endless lists of descriptors have led through rigorous concept and statistical analysis to genuinely useful psychological and clinical data, are expounded. In contrast to Paley's assertions, the study of caring in nursing to date has also produced information which is useful within nurse education and practice. CONCLUSION: There is no confusion concerning the things said about and the things done in the name of caring in our minds. We acknowledge that studying the actual phenomena of caring is difficult. However, in the absence of definitive descriptions of caring and precise methods to study it, the search for perfection has not paralysed action. Much has been learned about caring and much remains to be learned. PMID- 12074758 TI - Response to: 'Barriers and bridges to care: voices of homeless female adolescent youth in Seattle, Washington, USA' by J. Ensign and A. Panke (2002) Journal of Advanced Nursing 37, 166-172. PMID- 12074759 TI - Response to: Heartcare an Internet-based information and support system for patient home recovery after coronary artery bypass graft (CABG) surgery' by P. Flatley Brennan, S.M.Moore, G.Bjornsdottir, J.Jones, C. Visovsky and M. Rogers (2001) Journal Of Advanced Nursing 35, 699-708. PMID- 12074763 TI - Continuing medical education in Turkey: recent developments. AB - BACKGROUND: The Turkish Association of Medicine founded a Continuing Medical Education Accreditation Committee in 1993 to evaluate and accredit scientific meetings and publications. The aims of this project were to raise the standards of meetings and to introduce compulsory revalidation and re-certification for physicians in Turkey. DISCUSSION: Since the year 1994, 2348 applications to the Continuing Medical Education board have been made (mostly for scientific meetings), and 95% of these applications have been accepted. Physicians received 139.014 credits during this time. This number is increasing every year. Meeting organisers' demand for such a kind of evaluation is increasing, because participants increasingly request it. SUMMARY: Efforts for revalidation and re certification of physicians have not been completely successful yet. In the near future the Co-ordination Council of Medical Speciality Societies is going to oblige member associations to establish speciality boards. This will be the first step to the conventional use of Continuing Medical Education credits in occupational evaluation. Time-limited re-certification of physicians is the principal goal of Turkish Medical Association. Efforts to implement this change in legislation are being made. PMID- 12074764 TI - Radioimmunotherapy with yttrium-90 ibritumomab tiuxetan for patients with relapsed CD20+ B-cell non-Hodgkin's lymphoma. AB - The clinical development and US Food and Drug Administration approval in 1997 of the monoclonal anti-CD20 antibody rituximab have been major treatment advances for patients with B-cell non-Hodgkin's lymphoma (NHL). Rituximab produces responses in approximately 50% of cases of relapsed, low grade NHL. Most of these responses are partial remissions; cure remains elusive. One way to enhance the effectiveness of monoclonal antibodies is to chelate radionuclides such as yttrium-90 ((90)Y) to the antibody. ( 90)Y is a high-energy, beta-emitting radioisotope that delivers most of its radiation over a path length of 2 to 5 mm. Therefore, the antibody delivers, or targets, the radiation only to CD20+ cells, sparing normal cells from the radiation. Ibritumomab is the murine anti-CD20 antibody that was engineered to develop the human chimeric antibody rituximab. Tiuxetan is a linker/chelator that is attached to the antibody to form ibritumomab tiuxetan (Zevalin; IDEC Pharmaceuticals, San Diego, CA). Zevalin can be reacted with (111)indium ((111)In) for imaging and (90)Y for therapy. Phase I studies of Zevalin have determined that patients with a baseline platelet count greater than 150,000 10(6)/L receive 0.4 mCi/kg. Patients with a platelet count of 100 to 149,000 10(6)/L should receive 0.3 mCi/kg. Zevalin has a higher overall response rate (ORR) than its cold antibody counterpart rituximab, as demonstrated in two separate clinical trials. The first trial (IDEC 106-04) randomized 143 rituximab-naive patients with relapsed NHL to receive rituximab or Zevalin. The ORR for Zevalin was 80% compared with 56% for rituximab (P = 0.002). The second trial (IDEC 106-06) tested the efficacy of Zevalin in patients who were rituxan refractory; the ORR was 74%. The main toxicity of Zevalin was reversible myelosuppression. These studies indicate that radiolabeled anti-CD20 antibodies can produce a higher ORR than rituximab. Single-dose Zevalin is another treatment alternative for patients with relapsed low grade NHL. It is well-tolerated even by older adults. The exact role of Zevalin in the therapy of NHL is undetermined. New studies are underway to explore whether patients can safely receive a second dose of Zevalin and to combine Zevalin with high-dose chemotherapy and stem cell rescue. The outcome of these studies will be helpful in deciding how best to integrate this new modality into the treatment paradigm of NHL. PMID- 12074765 TI - New treatments for Hodgkin's disease. AB - Four of five patients with Hodgkin's disease (HD) will be cured with modern treatment strategies, depending on stage and risk factor profile. In early stage favorable HD, cure rates are greater than 90% with extended field (EF) irradiation, the standard treatment. However, the concept of EF irradiation therapy is being abandoned by most study groups because of the recognition of fatal long-term effects, especially the high rates of second solid tumors. Newer approaches include mild chemotherapy combined with involved field (IF) irradiation to control occult disease. Combined modality is the treatment of choice in early stages unfavorable (intermediate) HD, in which EF irradiation is substituted by IF irradiation. In the last three decades, because of the high relapse rates (30%-50%) after first-line polychemotherapy, the standard regimens were often modified. However, until recently, these efforts could not change the relatively poor outcome for patients with advanced stage disease. The introduction of a new dose-intensified regimen (BEACOPP) has significantly improved the prognosis for patients with advanced HD. Patients who relapse after radiation therapy alone for early stage HD have satisfactory results with combination chemotherapy and are not considered as candidates for high-dose chemotherapy with autologous stem cell transplantation. For patients with relapsed HD after combination chemotherapy, the data support the use of high-dose chemotherapy with autologous stem cell transplantation. PMID- 12074766 TI - Burkitt's and Burkitt-like lymphoma. AB - Burkitt's and Burkitt-like lymphoma (BL/BLL) are aggressive B-cell malignancies with a high proliferative rate that may be fatal within months if not treated promptly. Furthermore, treatment of BL/BLL requires comprehensive supportive care to avoid disease-related complications such as acute renal failure secondary to tumor lysis syndrome. Improvements in our understanding of the biology of BL and BLL have led to more effective therapeutic protocols. Clinical trials have demonstrated that short duration, multi-agent, dose-intensive chemotherapy regimens combined with aggressive central nervous system therapy results in long term survival rates in children and young adults near 70% to 80%, whereas long term disease-free survival rates in older adults remains suboptimal at 15% to 25%. Outcomes in HIV-associated BL/BLL are improved because of more effective chemotherapy regimens and enhanced HIV care. Autologous bone marrow transplantation has proven feasible in many patient populations with BL/BLL and may lead to cure in selected patients. Improved therapeutic strategies are warranted, such as integrating agents such as monoclonal antibodies to combination dose-intensive chemotherapy. Moreover, further study into the molecular biology of BL/BLL with attention to the role of c-myc dysregulation is needed to help predict prognostic factors and for the development of molecular targeted therapies. Clinical trials remain critical to determine the most effective treatment regimens that will continue to improve cure rates in this aggressive but treatable disease. PMID- 12074767 TI - Vaccine therapies for non-Hodgkin's lymphoma. AB - Various clinical observations suggest that non-Hodgkin's lymphomas (NHLs), particularly those of low histologic grade, can be controlled by immunologic mechanisms. Although many effective therapies exist for the initial treatment of low grade lymphomas, none are curative and most have significant toxic side effects. Several promising lymphoma tumor antigen vaccines are being studied at medical centers throughout North America. I favor the scientific evaluation of a therapeutic strategy for follicular NHL that places immune-based therapies forward in the treatment algorithm to the initial therapeutic decision point. Active immunotherapies (therapeutic tumor vaccines) are instituted in tandem with initial cytoreductive chemotherapy, and followed by passive monoclonal antibody therapies. The tumor-specific idiotype vaccines are favored because of their demonstrated potential for clinical activity in numerous human studies and their lack of significant toxic side effects. Rituximab and other monoclonal antibodies directed at normal B-cell antigens are known to abrogate the host's ability to mount primary humoral immune responses, including antitumor antibodies evoked by tumor vaccines. Therefore, one should consider deferring the use of these agents until after an attempt at generating a host humoral antitumor response using investigational tumor vaccines. Chemotherapy regimens containing highly immunosuppressive agents (ie, fludarabine) or organ dose-limiting toxicities (ie, doxorubicin) may be best reserved for later in the disease course for those failing the more conservative approaches and for cases with adverse prognostic features. This strategy may give patients the greatest chance at prolonged remission or cure while minimizing acute and chronic toxicities, although its impact on overall survival has not been proven. Low grade NHLs remain the proving ground for this treatment philosophy. Hopefully, in the future, similar strategies may be applicable to NHLs of other grades and histologies. PMID- 12074768 TI - Carcinoid tumors. AB - Carcinoids are rare endocrine tumors that can develop in several organs in the body. Clinically, patients can have a wide spectrum of signs and symptoms that range from incidental findings of a polyp during endoscopy to the carcinoid syndrome characterized by severe flushing, diarrhea, abdominal cramping, and life threatening right-sided heart failure. Most carcinoid tumors are indolent but can metastasize to regional lymph nodes and to other organs, including the liver, bone, and the central nervous system. Treatment is determined by tumor location and by the presence of distant metastasis. Surgical resection of the tumor is advocated in patients with localized disease and can often be curative. Long acting somatostatin analogs, including octreotide, octreotide long-acting repeatable, and lanreotide prolonged release, are effective in providing symptom relief in patients with the carcinoid syndrome. Patients with metastatic disease to the liver that is refractory to somatostatin treatment should be considered for hepatic artery occlusion. Overall, 5- and 10-year survival rates in patients with metastatic disease are favorable, although tumors can be resistant to most forms of medical or surgical therapy. PMID- 12074769 TI - Aldosteronoma. AB - Aldosteronoma is a surgically curable cause of hypertension. Recent studies have found aldosteronomas to be a more common cause of hypertension than previously thought. At least 2% of patients with hypertension may have an aldosteronoma. More than 50% of these patients are normokalemic because of earlier diagnosis or milder disease, but still benefit from adrenalectomy. Patients with hypertension should be screened for possible primary hyperaldosteronism regardless of their serum potassium level. When used in conjunction with the appropriate laboratory tests, high-resolution computerized tomography scanning helps the surgeon to differentiate accurately between an adrenal adenoma and bilateral adrenal hyperplasia. Focused approach and laparoscopic resection are the norm for the surgical treatment of aldosteronoma. PMID- 12074770 TI - Multiple endocrine neoplasia type I. AB - Multiple endocrine neoplasia type I is a rare autosomal dominant disorder with many endocrine and nonendocrine manifestations. Hyperparathyroidism, islet cell tumors, and pituitary tumors are diagnosed most commonly in these patients. There is controversy regarding treatment of the different manifestations and screening modalities of this disorder because no large series has determined the best therapeutic approach. Our institution advocates early screening with biochemical and radiographic testing in patients with a definite predilection for this disorder. Patients with hyperparathyroidism should undergo early surgical intervention of at least three and a half glands combined with bilateral upper thymectomy through a cervical incision. Although the recurrence rate is high, disease-free survival can be for as long as 30 years. Pituitary tumors are predominantly prolactinomas and growth hormone-releasing tumors. Prolactinomas usually require pharmacologic therapy, whereas growth hormone-releasing tumors are treated surgically. Enteropancreatic tumors should be treated surgically or medically, depending on the hormone secreted. Insulinomas respond well to distal pancreatectomy, with enucleation of the tumor from the head and uncinate process of the pancreas. However, there is controversy regarding the surgical treatment of gastrinomas. Carcinoid tumors should be treated with early surgical intervention. Adrenal lesions are thought to occur in association with pancreatic lesions and commonly have an indolent course. Adrenocortical cancers are uncommon in patients with multiple endocrine neoplasia type I. Skin lesions can be excised when cosmetically unappealing. PMID- 12074771 TI - Follicular thyroid carcinoma. AB - Follicular carcinomas are rare thyroid malignancies that are difficult to diagnose preoperatively. Fine needle aspiration is an excellent diagnostic tool and should be the initial step in managing the solitary thyroid nodule. Follicular carcinoma cannot be diagnosed with certainty by cytologic features alone; the diagnosis rests on the histologic findings of blood vessel or tumor capsule invasion. Surgical resection is the primary option for treatment. The extent of thyroidectomy for optimal survival outcome has not been determined scientifically. The outcome is excellent in minimally invasive follicular carcinoma with lobectomy and isthmusectomy; it is difficult to argue that total thyroidectomy is necessary. In a low risk prognostic group, for tumors other than minimally invasive carcinoma, lobectomy and isthmusectomy or total thyroidectomy can be justified. However, if total thyroidectomy can be done safely with a minimum of complications, then it has definite advantages for staging, postoperative surveillance, treatment, and possibly a lower recurrence rate and better survival rate. For all patients at high risk of recurrence, total thyroidectomy is preferred. PMID- 12074772 TI - Nonfunctioning islet cell carcinoma of the pancreas. AB - Islet cell cancers of the pancreas may follow an indolent course. Unlike adenocarcinoma of the pancreas, which is considered inoperable for cure in the presence of metastatic disease, islet cell carcinoma of the pancreas in selected cases warrants an aggressive approach toward resection. Significant palliation may be obtained from resection of primary tumors despite the presence of liver metastases. Furthermore, resection or ablation of limited, metastatic disease in the liver may be indicated. Therapeutic decisions must consider the patient's symptoms, coexisting morbidities, and the extent of the disease. PMID- 12074773 TI - Efficacy of physical therapeutic modalities in women with proven bladder overactivity. AB - OBJECTIVE: To assess the efficacy of physiotherapeutic treatment modalities in women with proven bladder overactivity. METHODS: One hundred and twelve women received ambulatory urodynamics. Based on both urodynamic variables of ambulatory cystometry (ACM) and the micturition diary, the Detrusor Activity Index (DAI) for each patient was calculated. After randomization, 68 women with a DAI> or =0.50 were defined as having proven bladder overactivity. In a single blinded RCT patients were randomized over four treatment groups, i.e. lower urinary tract exercises (LUTE); office- and home-based functional electrostimulation (FES); office-based FES and LUTE; no treatment. Patients treated received nine treatment sessions, once weekly. The primary outcome variable was the DAI, measured before randomization and, as soon as possible within a maximum of 14 days after the end of the study period. RESULTS: Intention to treat analysis in the group of 68 patients showed a statistically significant decrease of DAI-scores in the FES group (p=0.032) in comparison with no treatment, while this decrease was not statistically significant in the LUTE group (p=0.105) and the LUTE+FES group (p=0.672). CONCLUSIONS: Our conclusion is that, based on the DAI, in the homogeneous set of 68 patients, only FES seemed to be effective. PMID- 12074774 TI - Long-term safety, tolerability and efficacy of extended-release tolterodine in the treatment of overactive bladder. AB - BACKGROUND: The objective of the present study was to examine the long-term safety, tolerability and efficacy of tolterodine extended-release (ER) in patients who had completed 12 weeks' treatment in a randomised, double-blind study comparing tolterodine ER 4 mg once daily (qd), tolterodine immediate release (IR) 2mg twice daily and placebo. METHODS: Of the 1377 patients completing the 12-week study, a total of 1077 (78%) chose to continue with 12 months' open-label treatment with tolterodine ER 4 mg once daily, irrespective of their previous treatment. Safety was assessed after 3, 6, 9 and 12 months' treatment in the study. Efficacy was evaluated from micturition diary variables and patients' perception of bladder condition and urgency following 3 and 12 months' treatment. RESULTS: 71% of patients completed the 12-month study. Tolterodine ER was safe and well tolerated. Adverse events of the general (14.5%), autonomic (13.2%), gastrointestinal (11.4%), respiratory (9.8%) and urinary (9.1%) systems were the most frequently reported. Dry mouth was the most common event, occurring in 12.9% of patients, and was generally mild in severity. Other adverse events occurred in less than 5% of patients. There was no increase in the frequency of adverse events with long-term relative to short-term treatment. The efficacy of tolterodine was maintained over the 12-month treatment period; relative to baseline there were reductions in the number of incontinence episodes per week (median change -83%) and micturitions per 24 hours (median change -21%) and an increase in volume voided (median change +25%) after 12 months' treatment. An improvement in patient perception of their bladder condition was found in 75% of patients completing the study, and 51% had an improvement in patient perception of urgency. CONCLUSIONS: Tolterodine ER 4mg qd displayed a favourable safety, tolerability and efficacy profile during 12 months' treatment of patients with overactive bladder. PMID- 12074775 TI - Efficacy of oral doxepin and piroxicam treatment for interstitial cystitis. AB - OBJECTIVE: To establish the efficacy of a multidrug oral treatment with the tricyclic antidepressant agent doxepin and the cyclooxygenase (COX) inhibitor piroxicam in patients with interstitial cystitis (IC), who had failed standard therapy in an open, prospective, nonrandomized study. METHODS: A total of 37 patients diagnosed with IC received 75 mg doxepin and 40 mg piroxicam daily. The treatment was termed DOXCAM. Effectiveness of therapy was assessed with frequency volume charts, an IC symptom score and with cystometry prior to treatment, 8 weeks after the start and 4 weeks after termination of drug treatment. RESULTS: Medication was not tolerated by five patients. Twenty-six of 32 patients have experienced virtual total remission of symptoms (81%) and six patients had significant relief (19%). DOXCAM treatment resulted in a significant percent decrease in pain (65% versus 21%). Daytime frequency decreased from 17.6+/-5.7 to 11.3+/-3.6 voids while nocturia did not improve significantly. Twenty-three of the 26 patients who became symptom free and four of the six patients who showed significant improvement had a return of symptoms after cessation of therapy. CONCLUSION: It is reasonable to consider oral treatment with DOXCAM in those patients who have failed first-line therapies. PMID- 12074776 TI - Health status and its correlates among Dutch community-dwelling older men with and without lower urogenital tract dysfunction. AB - OBJECTIVE: To study health status and its correlates in older men with and without lower urogenital tract dysfunction. METHODS: Cross-sectional population based study on 1688 men aged 50-78 years without bladder or prostate cancer, radical prostatectomy, neurogenic bladder dysfunction or a negative advice from their general practitioner. Data were collected through self-administered questionnaires, including Sickness Impact Profile (SIP, three domains), Inventory of Subjective Health (ISH), International Prostate Symptom Score (IPSS) and International Continence Society (ICS) Male Sex questionnaire, medication use, socio-economic and lifestyle factors. Additional information was collected by measurement of blood pressure, transrectal ultrasonography of the prostate and uroflowmetry. Four health status domains were analyzed using the ISH and three domains of the SIP. Lower urinary tract symptoms (LUTS) were categorised using IPSS, erectile and ejaculatory dysfunction were defined using the ICS questionnaire. RESULTS: All urogenital characteristics and parameters were related to at least two of the health status domains. Multivariate regression analyses yielded that LUTS and cardiac symptoms were associated with suboptimal scores of all four domains. Chronic obstructive pulmonary disease and drugs for abdominal symptoms were related to three domains; erectile and ejaculatory dysfunction, muskuloskeletal or psycho(ana)leptic drugs and marital status to two domains. CONCLUSIONS: The impact of LUTS on health status was equally important as the impact of cardiac symptoms. The impact of sexual dysfunction was smaller than expected. Longitudinal studies are needed to determine how health status and illnesses interact. PMID- 12074777 TI - Efficacy and safety of a novel combination of L-arginine glutamate and yohimbine hydrochloride: a new oral therapy for erectile dysfunction. AB - PURPOSE: The goal of this double-blind, placebo-controlled, three-way crossover, randomized clinical trial was to compare the efficacy and safety of the combination of 6g of L-arginine glutamate and 6 mg of yohimbine hydrochloride (AY) with that of 6 mg of yohimbine hydrochloride (YP) alone and that of placebo (PP) alone, for the treatment of erectile dysfunction (ED). MATERIALS AND METHODS: Forty-five patients were included in this study. During each of the 2 week, crossover periods, drug was administered orally, one to two hours before intended sexual intercourse. The primary endpoint was change in the Erectile Function Domain score of the International Index of Erectile Function (IIEF). The secondary endpoints were patient and investigator assessments of treatment success. RESULTS: At the end of each treatment period, the Erectile Function Domain scores for AY, YP and PP were 17.2+/-7.17, 15.4+/-6.49 and 14.1+/-6.56, respectively. The difference between AY and PP was statistically significant (p=0.006). When stratified according to baseline scores over 14, those patients with mild to moderate MED had a better Erectile Function Domain response to treatment (AY=22.2+/-4.99, YP=18.2+/-5.59, PP=16.9+/-6.91, respectively) than those with scores 14 and below (AY=12.4+/-5.48, YP=12.7+/-6.25, PP=11.4+/-5.02, respectively). Investigators' and patients' assessment of efficacy was significantly improved by YP over PP. CONCLUSIONS: This pilot study shows that the on-demand oral administration of the L-arginine glutamate 6g and 6 mg yohimbine combination is effective in improving erectile function in patients with mild to moderate ED. It appears to be a promising addition to first-line therapy for ED. PMID- 12074778 TI - Recurrent varicocele: role of antegrade sclerotherapy as first choice treatment. AB - OBJECTIVE: To underline the role of antegrade sclerotherapy as first choice treatment in recurrent varicocele. METHODS: In 53 patients, aged 11-38 years, observed over a 30-month period, antegrade sclerotherapy was carried out for the treatment of recurrent varicocele. In seven of these patients, varicocele was bilateral. The right varicocele, however, had not been previously detected and these patients were, therefore, submitted to simultaneous bilateral antegrade sclerotherapy. RESULTS: Antegrade sclerotherapy was feasible in all 53 patients (60 varicoceles) and no significant complications were observed. Unsuccessful results occurred in only two out of the 55 varicoceles observed over a minimum follow-up period of six months. CONCLUSIONS: Whilst antegrade sclerotherapy is recognized as a low-cost, safe and effective method in the management of varicocele, the success rate in recurrent varicocele is even greater than in primary varicocele. If, in the preceding treatment, the internal spermatic vein has presumably been occluded (percutaneous retrograde sclerotherapy, surgical or laparoscopic retroperitoneal ligation of the entire bundle), antegrade sclerotherapy should be considered the treatment of choice. PMID- 12074779 TI - Estimation of prostate cancer risk on the basis of total and free prostate specific antigen, prostate volume and digital rectal examination. AB - BACKGROUND AND OBJECTIVE: Approximately 70% of the men with an elevated serum prostate-specific antigen (PSA) identified in prostate cancer screening do not have prostate cancer. Other available diagnostic variables may be utilized to reduce the number of false positive PSA results, but few algorithms for calculation of the combined impact of multiple variables are available. The objective of this study was to establish nomograms showing the probability of detecting prostate cancer at biopsy on the basis of total PSA, and the percentage of free PSA in serum, prostate volume and digital rectal examination (DRE) findings. METHODS: In a randomized, population-based prostate cancer screening trial 10284 men aged 55-67 years were screened during 1996 and 1997 in two metropolitan areas in Finland. Results for men (n=758) with a serum PSA of 4-20 microg/l were used to establish the risk nomograms. Of these 200 (26%) had prostate cancer at biopsy. RESULTS: Prostate cancer probability depended most strongly on the percentage of free PSA. Total PSA, prostate volume, and DRE also contributed to prostate cancer probability, whereas age and family history of prostate cancer did not. More false positive PSA results could be eliminated by using the multivariate risk model rather than the percentage of free PSA (p<0.001) or PSA density (p=0.003) alone. CONCLUSIONS: Wide variation in probability of detecting prostate cancer among screened men with a serum PSA of 4 20 microg/l was observed. The nomograms established can be used to avoid or defer biopsy in men with a low prostate cancer probability in spite of a serum PSA level exceeding 4 microg/l. PMID- 12074780 TI - Clinical outcome of patients with lymph node positive prostate cancer after radical prostatectomy versus androgen deprivation. AB - OBJECTIVE(S): To compare the outcome of patients with stage D1 (TxN+M0) prostate cancer undergoing radical prostatectomy or androgen deprivation alone. PATIENTS AND METHODS: Eighty-two patients treated for lymph node positive prostate cancer were retrospectively analyzed for time to progression, tumor-specific and overall survival. Furthermore, subsequent tumor and treatment related morbidity requiring intervention including frequency and duration of associated hospital stays was recorded. RESULTS: The extent of lymph node metastasis was significantly lower in 50 patients undergoing radical prostatectomy (+/- early androgen deprivation) compared to 32 receiving androgen deprivation only. The treatment groups, however, did not differ with regard to other characteristics including age, comorbidity, stage, grade and preoperative PSA. Mean actuarial progression-free, and tumor-specific survival was significantly longer for the radical prostatectomy patients (36% and 47%, respectively at 10 years) compared to androgen deprivation (15% and 32%, respectively). The latter group required more secondary interventions resulting in more frequent and overall longer hospital stays. CONCLUSIONS: Patients undergoing radical prostatectomy for stage D1 prostate cancer possibly benefit with regard to the necessity for secondary interventions and, at least for limited (solitary) nodal disease, in terms of progression-free and tumor-specific survival. However, the latter observation may be biased by a larger extent of lymph node metastasis in the androgen deprivation group. PMID- 12074781 TI - Intravesical Bacillus Calmette-Guerin treatment improves patient survival in T1G3 bladder tumours. AB - OBJECTIVE: To study the clinical and pathological factors that affect recurrence, progression and survival in pT1G3 bladder tumours treated conservatively. MATERIAL AND METHODS: From January 1979 to December 1996, 80 patients were conservatively treated for pT1G3 bladder tumours. All patients were studied for potential prognostic factors such as: age, sex, previous tumour recurrence, tumour size, multiple tumours, carcinoma in situ, and intravesical instillations. A longitudinal, retrospective, observational and analytical study was conducted to evaluate four different types of events: recurrence, progression, overall survival, and disease-specific survival. The chi(2) (Fischer exact test) and student t tests were used to assess the prognostic value of the qualitative and quantitative variables. Estimations of the survival distributions were calculated according to the Kaplan-Meier method and compared with the Log rank test. Multivariate analysis of the data was performed with Cox proportional hazard models. RESULTS: Among the 80 patients, 67 (84%) were men and 13 (16%) were women, with median age of 65.5 years. The median tumour size was 20 mm, most had single tumour (58.8%) and carcinoma in situ was found in six patients (7.5%). Thirty patients were treated with transurethral resection (TUR) of the bladder tumour and 50 patients were treated with TUR followed by BCG. The two groups of patients were comparable and followed up during a median time of 61 and 65 months, respectively (p=0.454). Kaplan-Meier estimators and Log rank tests demonstrated that patients with TUR alone recurred (p<0.0001), progressed (p<0.040) and died (overall survival: p<0.009; disease-specific p<0.040) earlier than patients who received intravesical instillations of BCG. The results were confirmed with Cox models and odds-ratios are presented. CONCLUSION: In this study, BCG adjuvant immunotherapy was the only factor affecting recurrence, progression and survival. Conservative treatment using TUR followed by BCG may improve disease-specific survival. PMID- 12074782 TI - Testicular germ cell cancer despite previous local radiotherapy to the testis. AB - BACKGROUND: Testicular intraepithelial neoplasia (TIN, also carcinoma in situ of the testis) is the uniform precursor of testicular germ cell cancer. Local radiotherapy to the testis with dosages of 18-20 Gy has been found to safely eradicate TIN and germ cells, too. Thus, the general assumption is that the development of invasive germ cell tumours can be prevented by this radiotherapy. PATIENTS AND METHODS: Herein, we report two patients with one-sided testicular tumour and biopsy-proven contralateral TIN. Both of them developed germ cell neoplasms in the remaining testis although local radiotherapy with 20 Gy had been applied to the testis. RESULTS: One patient developed pure seminoma 7 years after completion of radiotherapy, the other developed a combined tumour consisting of embryonal carcinoma and seminoma after 5 years. Treatment consisted of orchiectomy in each of the cases. Histologically, both had TIN in the testicular tissue surrounding the new growths. CONCLUSIONS: Pathogenetically, a small fraction of radioresistent TIN cells overcoming irradiation and progressing to full-blown germ cell cancer in the later course may be the histogenetic clue to explain these unexpected events. Other explanations, though less probable, could be technical radiotherapeutic failure due to targeting problems and a pre existing radioresistent germ cell tumour in the irradiated testicle. PMID- 12074783 TI - Intratubular germ cell neoplasia of the testis: testicular intraepithelial neoplasia. AB - The observations of Skakkebaek and the evolution of the concept of intratubular germ cell neoplasia (or testicular intraepithelial neoplasia (TIN)) indicate that most, but not all, germ cell tumors of the testis evolve from a common neoplastic precursor lesion: intratubular germ cell neoplasia, unclassified type (IGCNU). It is defined as the presence of malignant germ cells within the seminiferous tubules. At 5 years about 50% of patients with a testicular biopsy positive for IGCNU have developed invasive germ cell tumors, and only a small fraction remain free of invasive tumors by 7 years. Orchiectomy is the treatment of choice in patients with unilateral IGCNU, and low-dose radiation is efficacious in patients with bilateral IGCNU (although sterility is certain). So far, there is only one published report of occurrence of two cases of germ cell cancer despite previous local radiotherapy to the testis. A recent study demonstrated an estimated risk of recurrent IGCNU following chemotherapy of 21% and 42% at 5 and 10 years, respectively. PMID- 12074784 TI - Bone mineral content and related biochemical variables in patients with ileal bladder substitution and colonic Indiana pouch. AB - OBJECTIVES: To evaluate the effects of ileal bladder substitution or colonic Indiana pouch on skeletal bone density and various biochemical parameters related to bone metabolism. PATIENTS AND METHODS: In 27 patients with urinary diversion and 14 controls with benign urologic disease, bone mineral density (BMD), assessed by dual-photon absorptiometry; serum electrolyte, creatinine, alkaline phosphatase and parathyroid hormone levels were determined, and capillary blood gas analysed. BMD was measured in the lumbar spine and the femur neck. The mean time since surgery was 33.6+/-10.1 months in 17 patients with an ileal bladder substitution and 56+/-9.1 months in 10 patients with a colonic Indiana pouch (p=0.001). RESULTS: Although BMD did not change in the colonic Indiana pouch group, it was reduced in the patients with ileal bladder substitution compared to control group. The mean pH value was not statistically significant different in the both groups from the control group (p>0.2). The mean base excess value reduced in the substitution group (p<0.01). While alkaline phosphatase levels increased in both groups compared to control group (p<0.05), the mean parathyroid hormone level decreased only in the patients with ileal bladder substitution (p<0.05). The other biochemical parameters were similar in patients and control subjects. There was a statistically significant correlation between the base excess values and BMD values of the patients in both groups. CONCLUSION: Although there is decreased BMD in patients with an ileal bladder substitute, there is no change in BMD in the patients with Indiana pouch. Alkaline phosphatase levels increased in both patient groups indicating increased bone turnover. PMID- 12074785 TI - Self-image and performance in children with nocturnal enuresis. AB - OBJECTIVE: To study the self-image of children between 8 and 12 years with nocturnal enuresis, and to correlate the findings with age, sex, clinical symptoms, primary versus secondary enuresis and treatment failures. METHODS: The study-group embraced 50 university hospital, and therefore selected therapy resistant nocturnal enuresis-children, 27 boys and 23 girls. The mean age was 9.8, which means children aged between 8 and 12 years. Children were classified into two age groups: I=8-9 years and II=9-12 years. A total of 41 out of 50 had primary nocturnal enuresis. The mean number of treatments before intake was 5.6, A=1-4, B=5-8, C=9-12. Seventy-seven children without nocturnal enuresis were included in the control group, 31 boys and 46 girls. The method we used to measure the perceived competence of the children on specific domains of their life was the Dutch translation and also validation of the "Self-Perception Profile for Children" by Harter. The testing was performed before and after therapy. RESULTS: Children with nocturnal enuresis have a significantly lower perceived competence than children without nocturnal enuresis, concerning physical appearance (p<0.05) and global self-esteem (p<0.01). There is a main effect of gender (p<0.01) and age (p<0.05) concerning scholastic skills. There is a correlation with the number of treatment failures. The more treatment failures, the lower the self-esteem. After successful treatment, there is an improvement of 'athletic competence' and 'global self-esteem', but it is not significant. CONCLUSION: Nocturnal enuresis has important negative effects on the self-image and performance of children. Perceived competence was lower in girls than in boys with enuresis, and it was significantly lower in the higher age than in the lower. Children with day-time and night-time incontinence have a significantly decreased perceived competence on scholastic skills compared to children with nocturnal problems only. Successful treatment tended to increase athletic competence and global self-esteem. PMID- 12074786 TI - DNA alterations in body fluids as molecular tumor markers for urological malignancies. AB - OBJECTIVES: DNA-based tumor markers are characterized by unique specificity rendering them an attractive target for molecular diagnosis of cancer in body fluids like blood serum/plasma and urine. Both cell-free tumor DNA circulating in plasma/serum and cellular tumor DNA are detectable by minimally invasive measures. METHODS: Three main detection methods, microsatellite analysis, mutation analysis in genomic or mitochondrial DNA and gene promoter hypermethylation analysis are applied. Detection of gene promoter hypermethylation by methylation-specific PCR enables the best methodical sensitivity requiring a ratio of tumor DNA within normal DNA of less than 1:1000. RESULTS/CONCLUSIONS: Tumor DNA derived from renal cell carcinoma, bladder cancer or prostate cancer is detectable in considerably more than 50% of plasma/serum samples and more than 70% of urine samples from these patients. Because the targeted DNA alterations are absent or very rare in controls, the specificity of DNA-based tumor detection methods reaches almost 100%. Although the methodology currently is experimental, automatization will make it easier and less expensive. This review is focused on the potential clinical value of DNA-based analysis of body fluids for the initial diagnosis and the follow-up of urologic cancer patients. PMID- 12074787 TI - Chorionic gonadotropin beta-subunit and core fragment in bladder cancer: mRNA and protein expression in urine, serum and tissue. AB - OBJECTIVES: Many transitional cell carcinomas (TCC) of the bladder express the beta-subunit (CGbeta) of chorionic gonadotropin (CG), and elevated serum levels occur especially in advanced disease. We have compared the diagnostic utility of various methods for detecting CG and CGbeta expression at the protein and mRNA level. METHODS: We used RT-PCR to detect CGbeta mRNA in urinary cells and highly sensitive immunoassays to determine CG and CGbeta in serum and the core fragment of CGbeta (CGbetacf) in urine from patients under follow-up for bladder cancer. Tissue expression was studied by immunohistochemistry. RESULTS: CGbeta mRNA was detected in urinary cells in 50% (n=84) of the cancer cases and in none of the healthy controls (n=15). Positive staining for CGbeta in tissue samples was observed not only in 30% (n=96) of the TCC cases, but also in 5 of 20 histologically benign samples from TCC patients, and in 10 of 21 samples from benign bladder diseases. Serum and urinary concentrations of CGbeta were elevated in 29% (n=66) and 8% (n=72), respectively, while serum CG was elevated in 18% of the TCC patients. Urinary CGbetacf concentrations were higher in invasive (T1-T4) than superficial (T in situ and Ta) tumors (p=0.037), in cases positive for CGbeta mRNA (p=0.03) and cases with suspicious or malignant urinary cytology (p=0.002). The ratio of urinary to serum concentration of CGbeta showed the strongest correlation with tumor stage (p<0.00001), grade (p<0.00001), and staining for CGbeta (p=0.019). CONCLUSIONS: Although CGbeta expression may occur in benign bladder epithelium, CGbeta mRNA in urinary cells is a potential marker of bladder cancer. Urinary and serum CGbeta have low sensitivity in early disease, but the urine/serum ratio appears to indicate local release of CGbeta into urine. Further studies are needed to evaluate the clinical usefulness of different forms of CGbeta expression. PMID- 12074789 TI - Nosocomial urinary tract infections in urologic patients: assessment of a prospective surveillance program including 10,000 patients. AB - OBJECTIVE: Hospital-acquired urinary tract infections (HUTI) represent a significant impairment in the quality of health care. Incidence in catheterized patients has been estimated at approximately 20%, however few data are available in urologic patients. We report a prospective surveillance program over 6 years in our urologic department and evaluate its evolution. METHODS: Population consists of all patients admitted to the urology ward for 48 hours or more over a 6-year period from 1994. Data recorded: age, gender, duration of stay, insertion and removal of catheters, diagnosis of HUTI. ANALYSIS: calculation of incidence, and incidence density for HUTI and for catheter-related HUTI, analysis of trends by chi(2) trend test. RESULTS: A total of 10,054 consecutive patients were included, 52% were catheterized. The median incidence of catheter-related HUTI in catheterized patients was 13.0%, the incidence density was 25.1 HUTI/1000 patient days of catheterization. The proportion of HUTI and specific catheter-related HUTI patients decreased, respectively from 8.4% and 14.2% to 6.5% and 12.3% during the study period (p<0.05). CONCLUSION: The rate of HUTI was not as high as previously reported, perhaps due to a controlled catheter policy. Surveillance was associated with a significant decrease in infection rates, suggesting a beneficial feedback effect. Evaluation of diagnoses and surgical procedures would ensure an optimal quality control program. PMID- 12074790 TI - The urologist view of BPH progression: results of an international survey. AB - OBJECTIVES: To assess the opinion amongst the international urological community on whether benign prostatic hyperplasia (BPH) is a progressive disease. METHODS: A 15-item questionnaire was distributed to the mailing list of the publication European Urology Today as well as being accessed through the website of the European Association of Urology (EAU). The survey included questions on: whether BPH was a progressive disorder; what evidence there was to support this, including clinical parameters that could be identified; risk factors for progression; and prevention of progression. RESULTS: A total of 472 completed questionnaires from 47 countries were evaluated. Overall, the urologists surveyed agreed that BPH was a progressive disease, and that some patients were more prone to this than others. Increased post-void residual urine, reduced flow rate, increased urinary symptoms and urodynamic evidence of obstruction were considered to be the most significant risk factors for progression. Half of the urologists surveyed considered that current medical therapies could prevent progression of BPH, while a significant proportion thought that surgery was an effective preventative measure. CONCLUSIONS: BPH, although a benign disease, has a considerable impact on the patient's quality of life. Its progressive nature and the identification of risk factors for progression warrant further investigation. In the future, there is likely to be a shift in the paradigm from treatment of BPH complications to include the prevention of BPH progression. PMID- 12074791 TI - Comparison of a phytotherapeutic agent (Permixon) with an alpha-blocker (Tamsulosin) in the treatment of benign prostatic hyperplasia: a 1-year randomized international study. AB - OBJECTIVE: While the lipido-sterolic extract of Serenoa repens (LSESr) Permixon((R))-has been shown to have an equivalent efficacy to finasteride in patients with benign prostatic hyperplasia (BPH), to date, there has been no valid comparison of phytotherapy with alpha-blockers. The aim of this study was to assess the equivalent efficacy of Permixon and tamsulosin. METHODS: Eight hundred and eleven men with symptomatic BPH (I-PSS> or =10) were recruited in 11 European countries for a 12-month, double-blind randomized trial. After a 4-week run-in period, 704 patients were randomly assigned to either tamsulosin 0.4mg/day (N=354) or Permixon 320mg/day (N=350). I-PSS, QoL and Q(max) were evaluated at baseline and periodically for 1 year. Prostate volume and serum prostate-specific antigen (PSA) were measured at selection and at endpoint. The endpoint analysis was performed on the per-protocol population of 542 patients (tamsulosin: N=273; Permixon: N=269). RESULTS: At 12 months, I-PSS decreased by 4.4in each group and no differences were observed in either irritative or obstructive symptom improvements. The increase in Q(max) was similar in both treatment groups (1.8ml/s Permixon, 1.9ml/s tamsulosin). PSA remained stable while prostate volume decreased slightly in the Permixon-treated patients. The two compounds were well tolerated, however, ejaculation disorders occurred more frequently in the tamsulosin group. CONCLUSION: This study demonstrates that Permixon and tamsulosin are equivalent in the medical treatment of lower urinary tract symptoms in men with BPH, during and up to 12 months of therapy. PMID- 12074792 TI - Bilateral fine-needle administered local anaesthetic nerve block for pain control during TRUS-guided multi-core prostate biopsy: a prospective randomised trial. AB - INTRODUCTION: Transrectal multi-core biopsies of the prostate can cause substantial discomfort with the need for high dose systemic analgesics. In a prospective randomised trial we investigated the efficacy of fine-needle administered local anaesthesia for bilateral prostatic nerve block prior to transrectal ultrasound (TRUS) guided prostate biopsy. MATERIALS AND METHODS: One hundred and eight men suspected of having cancer of the prostate were randomised to receive TRUS-guided bilateral prostate nerve block prior to biopsy or not, when having no history of previous prostate biopsies (groups I and II, n=68). In group III (n=40) all patients with history of previous biopsies exclusively received local anaesthesia injection. Patients routinely underwent a 10-core biopsy regimen. For repeat biopsy 12 cores were taken. The consented patients documented pain on a visual analogue pain score. RESULTS: In the randomised groups I and II average pain score was 1.85 with versus 3.29 without periprostatic nerve block (p<0.0001). In group III the difference in pain stated for the present biopsy with local anaesthesia nerve block in comparison to the pain experienced with the previous biopsy solely under transrectal lidocaine gel was even higher (1.71 versus 4.59; p<0.0001). Pain relief was independent of the number of biopsy cores sampled. Overall cancer detection rate was 46% (50/108). CONCLUSION: Bilateral local anaesthesia nerve block prior to multi-core TRUS guided prostate biopsy significantly reduces pain independent of the number of cores taken. PMID- 12074793 TI - Conformal photon-beam radiotherapy of prostate carcinoma. AB - Three-dimensional conformal radiotherapy is the recommended radiation technique for localized or locally advanced prostate cancer. In the past decades, external beam irradiation procedures have evolved in the context of technical developments of radiation and imaging equipment. The article summarizes these developments and gives a definition of new techniques and their potential advantages over conventional irradiation. It is meant to provide urologists and medical and radiation oncologists with a better comprehension of modern radiation treatment of prostate cancer and its possible improvements in the future. PMID- 12074794 TI - Variability in the recurrence rate at first follow-up cystoscopy after TUR in stage Ta T1 transitional cell carcinoma of the bladder: a combined analysis of seven EORTC studies. AB - OBJECTIVES: To assess the variability between institutions in the recurrence rate at the first follow-up cystoscopy (RR-FFC) after transurethral resection (TUR) in patients with stage Ta T1 bladder cancer. METHODS: A total of 2410 patients from seven EORTC phase III trials conducted between 1979 and 1989 were included. Patients with single and with multiple tumors were analyzed separately according to whether or not they received adjuvant intravesical treatment. RESULTS: The RR FFC varied greatly between institutions. For patients with a single tumor, it ranged from 3.4% to 20.6% for patients not receiving any intravesical adjuvant treatment and from 0% to 15.4% in those receiving it. In patients with multiple tumors who had adjuvant treatment, it varied between 7.4% and 45.8%. There was a slight decrease over time in the recurrence rate for patients with single tumors, particularly in those receiving adjuvant intravesical treatment. CONCLUSIONS: For both patients with single and with multiple tumors, the percentage of patients with a recurrence in the bladder at the first follow-up cystoscopy after TUR varies substantially between institutions and cannot be explained by the factors that were assessed. It is suggested that the quality of the TUR performed by the individual surgeons may be responsible. PMID- 12074795 TI - Microsatellite alterations in human bladder cancer: detection of tumor cells in urine sediment and tumor tissue. AB - OBJECTIVES: Bladder cancer is the result of clonal expansion of cancer cells in which multiple genetic alterations have occurred. Loss of heterozygosity (LOH) studies have demonstrated that alterations in microsatellite regions are common in bladder cancer. This observation offers the possibility of early tumor detection by examining the DNA of urinary sediment. METHODS: We investigated alterations of 17 microsatellite loci in urinary bladder carcinomas of different stages and grades. Per locus, 19-30 specimens were evaluated. DNA was isolated from tumor specimens, urinary sediment and peripheral blood lymphocytes. DNA fragments of 17 microsatellite loci were amplified by PCR and analyzed for genomic alterations. RESULTS: Microsatellite alterations were detected in tumor tissue and urine sediment from 27 out of 31 patients (87%). Urine sediment analysis alone proved positive in 24 out of 31 patients (77%). The type of lesions most frequently detected was LOH (74% of all alterations), followed by length alteration (24%) and additional alleles (2%). On average, the alteration frequency was 22% per locus. The loci at chromosomes 9 and 18 proved most informative. No alterations were found in grade I tumors. The study revealed a correlation between microsatellite alterations and the respective grades and stages of the tumors. Average alteration frequencies per locus were: 27.4% in grade III versus 19.3% in grade II tumors, 26.5% in invasive versus 12.3% in superficial tumors. CONCLUSIONS: Our results demonstrate that microsatellite alterations are common in bladder cancer and that analysis of genomic instabilities in urine samples should be further evaluated as a method for bladder cancer screening in a high-risk group. Especially, when a set of microsatellites is used that shows a high probability of detecting alterations and allows easy handling, this could be an alternative or a completion to currently available methods. PMID- 12074796 TI - Retroperitoneoscopic partial nephrectomy using microwave coagulation for small renal tumors. AB - OBJECTIVES: The outcome of laparoscopic partial nephrectomy using a microwave tissue coagulator for treatment of small renal tumors was studied. PATIENTS AND METHODS: From June 1999 to May 2001, eight patients with small renal tumors of less than 5.0cm in diameter (1.0-5.0cm, T1N0M0) underwent retroperitoneoscopic partial nephrectomy. To control bleeding during the partial nephrectomy, the renal parenchyma around the tumor was coagulated using a microwave tissue coagulator with a needle of 1.5cm length. The tumor was circumscribed within the coagulated area with 8-13 punctures of the coagulation needle, and partial nephrectomy was performed using scissors and bipolar forceps. RESULTS: All eight patients successfully underwent the procedure retroperitoneoscopically. The average operative time was 295 minutes and the average blood loss was 129ml. Three patients showed urine leakage from the renal calyces, which was controlled by suturing retroperitoneoscopically. In two patients, the surgical margin was revealed to be positive for renal cell carcinoma by frozen section pathology and additional resection was performed in these individuals. The patients were discharged from the hospital with almost full convalescence on day 10 on average. Within the mean follow-up period of 10.4 months, no recurrence was found when examined with computer tomography (CT) using contrast media. As a complication, one patient experienced a decrease in function of the operated kidney caused by unknown reason. CONCLUSION: Retroperitoneal partial nephrectomy using a microwave tissue coagulator is useful for treatment of small renal tumors located at the peripheral area of the kidney. Bleeding from the renal parenchymal incision site is well controlled without occlusion of the renal artery with additional use of a bipolar coagulator, when necessary. Further experience and long-term follow-up are mandatory however, to establish the usefulness of this technique. PMID- 12074797 TI - Long-term follow-up after surgery for advanced colorectal carcinoma involving the urogenital tract. AB - OBJECTIVES: Advanced colorectal carcinomas frequently involve the urogenital tract. In the following we evaluate the long-term survival after radical surgical extirpation and the prognostic significance of involvement of specific urological organs. METHODS: Between January 1985 and April 1996, 101 patients underwent interdisciplinary tumour extirpation of an advanced colorectal carcinoma involving the urogenital tract. RESULTS: Of 68 men and 33 women, 40 presented with primary and 61 with recurrent carcinoma. As far as urological organs are concerned, the ureter was removed in 82 patients, followed by bladder (n=52), seminal vesicles (n=25), prostate (n=22), kidney, testicle and penis. Histology revealed cancerous infiltration in 52% of the organs resected. A negative surgical margin was obtained in 54% of the patients, 43% showed positive lymph nodes. There was a 41% peri-operative complication with a mortality rate of 5%. Five year overall survival was 24.4% (median 23 months) with prognostic factors being type of tumour (primary versus recurrent), surgical margin and lymph node status. Stratification according to these factors showed removal of bladder and prostate to be a favourable and ureteral removal to be an omnious factor. CONCLUSION: We conclude that multivisceral extirpation of advanced colorectal carcinomas involving the urogenital tract should be recommended in selected patients. Our data showed it to be a safe surgical procedure, which is associated with favourable long-term outcome in non-metastatic patients in whom complete surgical resection could be achieved. PMID- 12074798 TI - Peyronie's disease: surgical correction of 40 patients with relaxing incision and duramater graft. AB - OBJECTIVE: Assessment of a long-term follow-up of a previously described technique to correct the curvature of the penis in Peyronie's disease. MATERIAL AND METHODS: Forty patients (43-69 years old) were operated between 1989 and 1999. The eligibility criteria for surgery were: lesion stabilised during, at least, one year; angulation of 40 degrees or more; and difficult or impossible sexual intercourse. Moreover, the patients had to have good libido, maintained erections and interested sexual partners. The operative technique consisted in making an I-shaped relaxing incision in the zone of higher angulation of the tunica albuginea and the defect created in the tunica was covered with a human duramater graft. RESULTS: The follow-up varied from 1 to 6 years (70%, > or =2 years). Cosmetic results: 95% good, 5% satisfactory (not needing further correction). Functional results: 85% good, 15% erectile dysfunction (10% moderate and 5% severe). COMPLICATIONS: one recurrence, one partial necrosis of the foreskin, nine decreases in the sensibility of the glans (eight recovered after 4 12 months post-operatively). CONCLUSIONS: The I-shaped relaxing incision and duramater graft procedure had a satisfactory success rate in the correction of severe Peyronie's disease. The utilisation of ready to use material (homo or hetero graft) avoids additional surgery, reduces the operative time and possible complications. Like in other procedures, the major problem of this surgery is the erectile dysfunction rate. PMID- 12074799 TI - Effects of pelvic floor exercises in middle aged women with a history of naive urinary incontinence: a population based study. AB - OBJECTIVE: To follow the effect of a 4-month pelvic floor exercise (PFE) program in women with naive urinary incontinence with vaginal electromyography (EMG), pressure and palpation and also to compare the initial findings with symptom-free women of the same age. METHODS: The pelvic floor function expressed with vaginal EMG, pressure and palpation was measured before, during and after 4 months of PFEs in 60 previously untreated incontinent women, 50 of whom completed the study. The patients' perception of the situation and the amount of leakage were estimated before and after PFE. The incontinent group was compared at baseline and after PFE with 28 healthy controls. All women in this study (age 53-63) were randomly recruited from a major population based study. RESULTS: At baseline, the incontinent women had significant reductions of both vaginal EMG activity and pelvic floor muscle condition as estimated by palpation compared to the healthy group. During training a successive, significant increase was seen in both EMG, pressure and palpation and the values eventually exceeded those of the healthy women. The measures reflecting improvement of pelvic floor function thus showed a consistent and progressive pattern. The degree of improvement was higher in those with initial high values in the muscle function tests than in the women with lower initial EMG values, pressures and findings on palpation. No differences were seen between patients with a history of stress incontinence and patients with an urge component, i.e. urge or mixed incontinence. Sixty-four percent of the women were satisfied and wanted no further treatment. The median leakage at pad-test decreased from 5 (range 0-328) to 1 (range 0-126) g/24h. The correlation between the vaginal and the pad-test measurements was weak. CONCLUSION: Women with urinary incontinence have a significant reduction of pelvic floor function as estimated with vaginal EMG and palpation as compared to symptom-free controls. A successive normalization of vaginal EMG, pressure and findings at palpation was seen during the 4-month training period. Incontinence of both stress type and with an urge component can be alleviated in most of the women with PFE. These methods might be useful for routine evaluation of the pelvic neuromuscular disorder present in incontinent women. PMID- 12074800 TI - Alterations of cortical electrical activity in patients with sacral neuromodulator. AB - OBJECTIVES: Sacral neuromodulation represents chronic stimulation of the sacral (S3) nerve. So far, the mode of action and neuro-anatomical basis is unclear. Sacral reflex mechanisms as well as pontine or cortical centers of modulation have been postulated. Our aim was to evaluate possible alterations in electroencephalogram (EEG) activity as an indicator of a supraspinally mediated mechanism of sacral neuromodulation. MATERIALS AND METHODS: We analyzed serial EEGs (apparatus: Kolner Vitaport System) using electrodes placed at Fz, Cz, Cz' and Pz in 10 patients. Subsequently, the sacral (S3) nerve was stimulated by means of an impulse generator (Medtronic, Interstim 3023) using an on-off paradigm with a 1.5s "on" interval followed by a 10s stimulation break. Raw data were analyzed using both Matlab 4.0 software and a specially developed averaging routine. RESULTS: All patients demonstrated a cortical potential complex following sacral root stimulation with an early electronegative component at 50 ms with a mean amplitude of 23 microV followed by a late potential component with a mean latency of 253 ms and a mean amplitude of 5 microV, both with a maximum at Cz, corresponding to the post-central gyrus. This finding occurred irrespective of patient's reports of actually feeling the neuromodulator being switched on and off. CONCLUSION: In neuromodulation responders, both short and long latency cortical potentials can be reproduced with a maximum at the sensory cortical area. Although these potentials are similar to cognitively mediated "event related potentials", they are clearly distinct from any subjective sensory or even painful response since several patients of this series have not been able to feel any neuromodulator action. Therefore, this pilot study indicates a supraspinally mediated site of modulation, most probably in sensory cortex areas. PMID- 12074801 TI - Estrogens affect endothelin-1 mRNA expression in LNCaP human prostate carcinoma cells. AB - OBJECTIVE: To study effects of estrogens on endothelin-1 (ET-1) mRNA expression in the androgen-sensitive LNCaP-FGC cell line and its androgen-resistant derivative LNCaP-r. Further, if effects of estrone sulfate (E1S) are mediated via conversion to estradiol-17beta (E2). Estrogens have been shown to down-regulate ET-1, a mediator of the osteoblastic response of bone to metastatic prostate cancer. METHODS: Cells were grown in steroid-depleted medium and incubated for 2 4 and 48 hours with 0, 1, 10, and 100 nM of either E1S or E2. mRNA levels were measured with an RT-PCR technique. Estrogen metabolism by LNCaP-FGC cells was studied by incubation with estrone (E1) and E1S at the same conditions, followed by determination of E1 and E2. RESULTS: ET-1 mRNA expression in LNCaP-FGC cells was significantly suppressed by E2 and E1S following incubation for 2-4h but after 48 h only by E2 at 1 and 10nM and in LNCaP-r cells only by E2 at 100 nM following 2-4h of incubation. ET-1 mRNA expression was significantly higher in untreated LNCaP-r than in untreated LNCaP-FGC cells. E1 was efficiently transformed into E2 by LNCaP-FGC cells but very little to E1 and no E2 was formed from E1S. CONCLUSION: ET-1 mRNA expression in LNCaP-FGC can be inhibited by E2, but also by its prehormone E1S. The lack of formation of E2 from E1S suggests a mode of action not related to classical steroid receptors. The higher level of ET 1 mRNA expression found in LNCaP-r cells may reflect the capability of a hormone refractory tumor to maintain activity on its own, independently of known regulatory mechanisms such as sex steroids. PMID- 12074803 TI - Proximal hypospadias. PMID- 12074804 TI - Modern approach of diagnosis and management of acute flank pain: review of all imaging modalities. AB - Acute flank pain is a common and complex clinical problem which might be caused by a variety of urinary and extraurinary abnormalities among which ureterolithiasis being the most frequent cause. Plain abdominal radiographs combined with intravenous urography (IVU) have been the standard imaging procedures of choice for the evaluation of acute flank pain over the last decades. Direct detection of even small ureteral calculi is achieved in 40-60%, whereas using indirect signs such as ureteral and renal pelvic dilatation stone detection is possible in up to 80-90% of all cases. However, IVU might be hampered by poor quality due to lack of bowel preparation, by nephrotoxicity of contrast agents, by serious allergic and anaphylactic reactions in 10% and 1% of the patients, respectively, and by significant radiation exposure. The use of ultrasonography (US) in the management of acute flank pain has been growing and when combining the findings of pyeloureteral dilatation, direct visualization of stones, and the absence of ureteral ejaculation, the sensitivity to detect ureteral dilatation can be as high as 96%. Recently, unenhanced helical CT (UHCT) has been introduced as imaging modality with a high sensitivity and specificity for the evaluation of acute flank pain. UHCT has been demonstrated to be superior since (1) it detects ureteral stones with a sensitivity and specificity from 98% to 100% regardless of size, location and chemical composition, (2) it identifies extraurinary causes of flank pain in about one third of all patients presenting with acute flank pain, (3) it does not need contrast agent, and (4) it is a time saving imaging technique being performed within 5min. Based on the data published, one can predict that UHCT will become the imaging procedure of choice for evaluation of acute flank pain within the next years. The purpose of this review is to critically evaluate the role all imaging modalities available for a modern approach of diagnosis and management of acute flank pain with regard to their sensitivity, specificity, positive and negative predictive values and their complications, toxicicty and morbidity. PMID- 12074805 TI - Prognostic factors in localised prostate cancer with emphasis on the application of molecular techniques. AB - Prostate cancer is the most prevalent malignancy in males in the Western world and the second leading cause of male cancer death. Prostate specific antigen (PSA) based screening and case finding leads to identification of early stage prostate cancer. It is often difficult to discriminate between patients that need curative treatment and those that can be managed conservatively. Prognostic factors are used to make this clinical decision. Based on the classification proposed by the American College of Pathologists and the World Health Organisation, selected prognostic factors in prostate cancer are described. Clinical applicable factors are stage, grade and serum PSA. Prognostic factors that are not routinely used (for various reasons) are ploidy, histological type and cancer volume in needle biopsies. All other factors (including circulating tumour cells, angiogenesis, growth factors, proliferation rate, apoptosis, nuclear morphometry, neuroendocrine differentiation, loss of chromosomal regions, tumour suppresser genes and adhesion molecules) are promising as prognostic factor although currently their use in clinical decisions is not recommended. The role of these factors in prostate cancer growth and their predictive value are discussed. The rapid developments in molecular techniques allow assessment of structure or function of thousands of genes in a prostate biopsy sample. We expect that molecular characterisation of tumour material will become a clinically important tool to predict prognosis in patients with localised prostate cancer. PMID- 12074806 TI - Molecular aspects of bladder cancer IV: gene therapy of bladder cancer. AB - Cancer gene therapy is the applied consequence of the intense research on cell function during the last decades. With the discovery of genes, the genetic code and gene functions many diseases, like bladder cancer, were linked to dysfunction of the cell's genetic material. Soon the wish for a direct treatment of the underlying cause of such genetically based diseases, a "gene therapy", arose and the first successful attempt in 1990 jumpstarted the development of gene therapies [Hum. Gene Ther. 4 (1993) 521], especially for cancer. To date treatment of bladder cancer remains a challenge to physicians. In spite of advances in diagnose and treatment over the last decades recurrence and progression rates remain high, especially in superficial bladder cancer. Gene therapy may provide the yet missing additional treatment to finally achieve a reduction of recurrence and progression. The variety of gene therapy strategies for bladder cancer developed or under investigation reflects the desire for a further treatment option for bladder cancer. This review intends to explain general strategies and state of the art approaches in bladder cancer gene therapy, highlight recent advances and give an outlook on what to expect from gene therapy in future. PMID- 12074807 TI - Efficacy and safety of fixed-dose oral sildenafil in the treatment of sexual dysfunction in depressed patients with idiopathic Parkinson's disease. AB - OBJECTIVE AND METHODS: The efficacy and safety of oral Sildenafil, a potent inhibitor of phosphodiesterase type 5, were evaluated in depressed men with idiopathic Parkinson's disease and erectile dysfunction. Thirty-three men were enrolled in a 4-month prospective, open-label, fixed-dose study, and received 50mg of Sildenafil in the home setting approximately 1 hour before sexual activity, not more than once daily. Efficacy was determined by responses to question 3 (ability to achieve an erection) and question 4 (ability to maintain an erection) of the 15-item International Index of Erectile Function (IIEF). Other measures of efficacy included the five sexual function domains of IIEF, a global efficacy question, the Beck Depression Inventory (BDI) and the Hamilton Depression Rating Scale-21 (HDRS-21). RESULTS: At the end of the study, improved erections were reported by 84.8% of patients. Sildenafil significantly increased patients' ability to achieve and maintain erections. Significant improvements were also observed in the IIEF domains for erectile function, orgasmic function, intercourse satisfaction and overall sexual satisfaction. BDI and HDRS scores improved from baseline to the end of the study. A clear improvement of depressive symptoms was observed in 75% of patients. Sildenafil was well tolerated in all the patients. CONCLUSIONS: Treatment with oral Sildenafil improves erectile function and, indirectly, depressive symptoms in patients with idiopathic Parkinson's disease stages 1-3, and is well tolerated. PMID- 12074808 TI - Switching from long-term treatment with self-injections to oral sildenafil in diabetic patients with severe erectile dysfunction. AB - OBJECTIVE: To assess the efficacy of oral sildenafil in diabetic men with severe erectile dysfunction (ED), who are successfully treated with intracavernous injections of vasoactive drugs. METHODS: 81 impotent diabetic men (29 with type 1 and 52 with type 2) were treated for 1-7 years with self-injections. 13 men were treated with 10 microg and 15 with 20 microg of prostaglandin E1 (PGE1), and 53 with a mixture of PGE1 20 microg and papaverine 7.5-40 mg (MIX). After a 1-week washout period, they changed to oral sildenafil in titrating doses up to 100mg. The change was successful if the man achieved an erection and penetration even once. RESULTS: Sildenafil was discontinued in 23 men because of insufficiency. Eleven men (13.6%), all with type 2 diabetes, responded to sildenafil (10 previously treated with 10 microg and 1 with 20 microg of PGE1, none treated with MIX). Thus, 39.2% of the treated with PGE1 responded to oral sildenafil. The response was influenced by the age, the type of diabetes (type 2) and the kind of the previously injected drug (PGE1 10 microg); it was not influenced by the duration of diabetes, ED and treatment with self-injections. CONCLUSIONS: Despite the well documented efficacy of sildenafil, self-injections continue to be the solely effective therapeutic modality in many diabetic men afflicted by severe ED. Only the younger men with non-insulin-dependent diabetes, treated with low doses of PGE1 are more likely to respond to oral sildenafil and change treatment. Men with insulin-dependent diabetes or treated with mixtures of vasoactive drugs are not likely to respond to oral sildenafil. PMID- 12074809 TI - Management of chronic testalgia by microsurgical testicular denervation. AB - OBJECTIVES: Chronic testicular pain (CTP) is defined as uni- or bilateral, intermittent or continuous testicular discomfort of at least 3 months duration that interferes with the patient's daily activities and prompts him to seek medical advice is a rather common urological manifestation of chronic pain syndrome. Diagnosis and treatment of CTP has been a difficult and often unrewarding clinical situation. Success rates of conservative and surgical measures including epididymectomy and orchiectomy rarely exceed 55-73% and 10 40%, respectively. We report our experience on microsurgical testicular denervation as therapeutic option in CTP. PATIENTS AND METHODS: Following an extensive preoperative work-up (urine/semen cultures, transrectal ultrasound, testicular sonography, pain and orthopedic consultation) not revealing any pathologic abnormalities and a positive response to spermatic cord block, 35 patients underwent microsurgical testicular denervation. In brief, spermatic cord was dissected, vas deferens, cremasteric muscle and testicular vessels were separated. After identification of the testicular artery by application of vasodilatating agents using magnifying loops or the operating microscope, all structures besides the testicular artery, vas deferens and 1-2 lymphatic vessels were coagulated and transsected using bipolar diathermy. RESULTS: After a mean follow-up of 31.5 months 34/35 (96%) patients are completely pain-free; no intra- or postoperative complications were encountered. No case of testicular atrophy or hydrocele formation was observed during postoperative follow-up. CONCLUSIONS: Microsurgical testicular denervation results in reliable and reproducible excellent therapeutic success rates of 96% and should be integrated in the management of CTP at an early stage. High success rates require adequate and meticulous diagnostic work-up of the patients by spermatic cord block using saline as placebo and different local anaesthetics as an initial therapeutic armentarium predicting postoperative outcome. PMID- 12074810 TI - Treatment of varicocele: a prospective randomized comparison of laparoscopy versus antegrade sclerotherapy. AB - OBJECTIVES: To compare the therapeutic success, the morbidity and the costs of antegrade sclerotherapy versus laparoscopic varicocelectomy. METHODS: Seventy-six consecutive varicocele patients were randomly assigned to two treatment arms. Preoperative and 3 month postoperative sperm density, motility and morphology were analysed. The diagnosis of the varicocele was established clinically and with Doppler ultrasonography. RESULTS: Fifty-eight patients treated by either of the two methods were followed up. The recurrence rate increased progressively with the size of the varicocele in both groups. The postoperative incidence of complications particularly hydrocele formation was significantly higher in the laparoscopic group. The costs of the disposable material for laparoscopic varicocelectomy was twice as high as for sclerotherapy. CONCLUSIONS: Antegrade sclerotherapy is the less invasive treatment method of male varicocele with lower costs and better outcome and should therefore be the preferred treatment method for male varicocele. PMID- 12074811 TI - Collagen fleece for defect coverage following plaque excision in patients with Peyronie's disease. AB - OBJECTIVES: Surgical correction of penile deviation in patients with Peyronie's disease by tunical plication often leads to shortening of the penis. It is, thus, recommendable to combine tunical plication with plaque incision or excision. The resulting tunical defect, however, requires grafting, and various techniques have been described. In comparison with tunical plication, all of these combined techniques are associated with increased operation time, mainly due to additional procedures necessary at the donor site, and may result in defects at the donor site. We here report a novel surgical technique by which tunical defects after partial excision of plaques are covered by a ready-to-use collagen fleece coated with tissue sealant (TachoComb). METHOD: A prospective clinical observation trial was conducted in 19 patients with penile deviation due to Peyronie's disease. RESULTS: In all patients, a reliable closure of the Tunica albuginea was achieved, and no postoperative haematoma formation was observed. Postoperatively, none of the patients suffered from erectile dysfunction. During the follow-up period of 25 months, objective and subjective improvement was 83% and 72%, respectively. CONCLUSION: The present data indicate that this novel surgical technique may be of benefit in patients with Peyronie's disease and should thus be further evaluated. PMID- 12074812 TI - Comparative investigations on the retrieval capabilities of various baskets and graspers in four ex vivo models. AB - OBJECTIVES: The increasing application of ureterorenoscopy for the treatment of urolithiasis has produced a myriad of different help-tools for stone retrieval. In this study, we compared the retrieval capabilities of different baskets and graspers in ex vivo models and attempted to find the most appropriate tool for stone extraction considering the location and size of stone, number of stones and potential harm to urinary tract tissue. METHODS: We created four different ex vivo models with porcine kidneys and ureters and natural human stones: (1) single ureteral stone (4mm); (2) single impacted ureteral stone (6mm); (3) Steinstrasse; (4) single lower-pole stone (5mm). With the aid of four baskets of different design (flat-wire basket Segura and Nitinol basket Zerotip, helical Gemini, and Parachute) and three graspers (two-prong, three-prong Tricep, and Nitinol grasper Graspit) we performed repeated stone extraction 10 times in each model. The time for complete stone removal was calculated. Macroscopical evaluation of tissue damage was performed after each series. In the Steinstrasse model the frequency of instrument reinsertion into the ureter was also calculated. RESULTS: In the single ureteral stone model, the fastest stone removal was achieved with the two prong graspers, three-prong graspers and helical basket (20, 26 and 31s, respectively). Segura and Parachute needed more time (55 and 86s, respectively). Impacted stones were removed fastest with the two- and three-prong graspers (38 and 52s, respectively), Segura and Gemini were slower (89 and 114s, respectively). The Steinstrasse was cleared fastest by the helical basket with the lowest frequency of endoscope reinsertion (66s, 1.4 reinsertion). The Segura, 149s with 3.2 reinsertion was needed for the same procedure. With the Parachute and two-prong graspers 163s, 1.8 reinsertion and 261s, 4.6 reinsertion were needed, respectively. During postprocedural macroscopical evaluation of uretral tissue, the Parachute basket and three-prong graspers demonstrated the highest risk of mucosal and muscle damage. In the lower-pole kidney model, the poor deflectability of the ureterorenoscope prevented access to the lower-pole with the two- and three-prong graspers. There was no significant difference in stone retrieval time between the remaining Nitinol basket Zerotip and Nitinol grasper Graspit (87 and 61s, respectively ). The number of failures was 4/10 for Zerotip and 5/10 for Graspit. No significant mucosal damage was noted with these tools. CONCLUSIONS: Our ex vivo models demonstrated that the design of graspers and baskets affects the time of stone retrieval in different situations and causes the various grades of tissue damage. PMID- 12074813 TI - "Quality of life" assessment of urination in elderly Japanese men and women with some medical problems using International Prostate Symptom Score and King's Health Questionnaire. AB - OBJECTIVES: To investigate the relationships among lower urinary tract symptoms (LUTS), satisfaction at urination, and quality of life (QoL) in elderly Japanese men and women, who attended physicians for their medical problems. METHODS: A total of 437 Japanese men and 196 women, aged 50 or older, responded to a questionnaire, which included seven questions regarding urination, one question about bothersomeness (QoL index) from the International Prostate Symptom Score (IPSS), two questions about frequency of urinary incontinence and 16 questions from the King's Health Questionnaire (KHQ). RESULTS: Of 401 patients who completed the IPSS and KHQ questionnaires, 235 (72.3%) men and 85 (64.7%) women had moderate to severe LUTS. In both genders, LUTS severity was significantly correlated with satisfaction status and KHQ-QoL domain scores. Satisfaction status also correlated with KHQ-QoL scores. Thirty-eight (13.5%) men and 27 (22.7%) women had frequent incontinence episodes. When excluding the patients with frequent incontinence episodes, KHQ-QoL scores of severe LUTS decreased in the domains of role, physical and social limitations, personal relationships and sleep/energy in women alone. CONCLUSIONS: Urination problems and related QoL status could be easily assessed by IPSS and QoL index in the elderly. Urinary incontinence appeared to be more associated with QoL in elderly women. The KHQ may be useful to assess the detailed urination-related QoL status of elderly men and women with LUTS or urinary incontinence. PMID- 12074814 TI - High dose rate brachytherapy of localized prostate cancer. AB - OBJECTIVE: We evaluated the safety and efficacy of high dose rate (HDR) brachytherapy using Iridium-192 (Ir 192) and 3D conformal external beam radiotherapy in patients with localized prostate cancer. METHODS: A total of 444 patients with localized prostate cancer underwent combined radiotherapy with interstitial Ir 192 and 3D conformal external beam radiotherapy between December 1992 and March 2001. The 230 patients, treated between December 1992 and December 1997 were analyzed. All patients underwent laparoscopic pelvic lymph node dissection to exclude patients with lymphatic involvement. Ir 192 was delivered twice with a 1-week interval in HDR remote control technique. The interstitial dose from December 1992 to December 1993 was 10Gy, after December 1993 the dose was reduced to 9Gy per treatment session. The interstitial application was followed by external beam radiation of 45Gy for cT1-cT2 and 50.4Gy for cT3 tumor (40Gy from December 1992 to December 1993). Progression was defined as biochemical failure according to ASTRO criteria, e.g. three consecutive PSA rises following the PSA nadir. RESULTS: The median PSA value decreased from 12.8 to 0.93ng/ml 12 months after treatment. Median PSA value was 0.47 after 24 months, 0.30ng/ml after 36 months and 0.18ng/ml after 60 months. 68% of the biopsies were negative 24 months after therapy. Progression-free rate was 100% for cT1 tumors, 75% for cT2 and 60% for stage-cT3 on 5-year follow-up. Five-year overall survival was 93%, 5-year disease-specific survival was 98%. Initial PSA value <10ng/ml, low stage and low grade were significantly related to 5-year progression-free survival. CONCLUSIONS: Combined HDR brachytherapy with Ir 192 is an alternative treatment option especially for patients with cT3 prostate cancer. Initial PSA value, stage and grade, are important prognostic factors. PMID- 12074815 TI - Complications following permanent prostate brachytherapy. AB - OBJECTIVES: The acute and chronic complications of permanent prostate brachytherapy are discussed. MATERIALS AND METHODS: Review of literature for the complications associated with iodine-125 (125I) and palladium-103 (103Pd) prostate brachytherapy. Acute complications included urinary retention, changes in the International Prostate Symptom Score (IPSS) and need for TURP. Chronic morbidity included permanent urinary symptoms, incontinence, radiation proctitis and erectile dysfunction. RESULTS: Urinary retention occurred in 1.5-22% of the patients postimplant. Acute urinary symptoms increased by over 100% 1 month after the procedure. By 12 months, the symptoms were either back to baseline or slightly elevated in over 90% of the patients. Significant obstructive symptoms or persistent urinary retention necessitating TURP occurred in 0-8.7%. Urinary incontinence was found in 0-19% treated by implant without associated TURP, in 0 85% for those who had a TURP prior to the implant and in 0-17% for those who had the TURP subsequent to the implant. Potency rates ranged from 34% to 86% 1-6 years postimplant. Radiation proctitis was found in 0.5-21.4%, with significant injury (fistula) occurring in 1-2.4%. CONCLUSIONS: The data from this report suggests that permanent prostate brachytherapy can be accomplished with minimal short- and long-term morbidity. Attention to detail as well as an appreciation to the causative factors for the morbidity will help reduce treatment-related side effects. PMID- 12074816 TI - Importance of post-implant dosimetry in permanent prostate brachytherapy. AB - OBJECTIVE: Post-implant dosimetry has become the gold standard for implant evaluation and it is recommended that it be performed on all patients undergoing prostate brachytherapy. The technique, results and correlation with clinical outcomes will be presented. METHODS: The method and outcomes of post-implant dosimetry are explored by outlining the experience at the Mount Sinai Medical Center, New York, as well as reviewing the literature. The most accurate time to perform post-implant dosimetry is 1 month after implant. Computed tomography (CT) based dosimetry is currently the best available technique for performing this analysis. The technique involves taking 3-mm abutting CT slices throughout the implanted area. The prostate and normal structures are outline on the CT slices. These structures are recreated in three dimensions. Dose volume histograms (DVH) are created and allow the dose to these organs to be quantified. RESULTS: The relationship between dosimetric findings and clinical outcomes has been established. The dose delivered to 90% of the prostate on DVH (D90) has been correlated to prostate-specific antigen (PCA) control and post-treatment biopsy results. D90 values of >or=140Gy have been associated with improved biochemical control and lower positive post-treatment biopsy results. Doses derived from the dosimetric analysis to prostate, urethra and rectum have been correlated with the development of acute and chronic urinary morbidity, sexual potency and rectal morbidity. Future initiatives involve performing dosimetric calculations intraoperatively at the time of the implant. CONCLUSIONS: Post-implant CT-based dosimetry is an essential component of prostate brachytherapy. It is the only method of assessing the actual dose delivered to the prostate and normal surrounding structures. Future development in post-implant and intraoperative dosimetry will continue to improve permanent prostate brachytherapy as a safe an effective treatment for prostate cancer. PMID- 12074817 TI - Disease specific survival as endpoint of outcome for bladder cancer patients following radical cystectomy. AB - OBJECTIVE: To directly compare disease specific and overall survival as endpoints in the outcome analysis of a large number of cystectomy patients and to define predictors for survival. MATERIALS AND METHODS: We retrospectively analyzed the records of 686 patients who underwent bilateral pelvic lymph node dissection (PLND) and radical cystectomy from 1980 to 1990 at Memorial Sloan-Kettering Cancer Center. RESULTS: Disease specific survival characterized a clearly more favorable patient outcome than overall survival in the entire patient population as well as patients with organ confined (OC) and non-organ-confined disease (NOC): 10-year disease specific and overall survival rates for patients with OC disease (or=P3b) were 72.9% versus 49.1%, 61.7% versus 40.8% and 33.3% versus 22.8%, respectively. In node positive (N+) patients 10-year disease specific and overall survival rates were 27.7% and 20.9%, respectively. In a multivariate analysis organ confinement and nodal status were the strongest independent predictors of disease specific survival in all patient categories. However, stratification according to organ confinement and nodal status revealed additional prognostic parameters. CONCLUSION: Organ-confined bladder cancer translates into high disease specific survival rates following radical cystectomy. Outcome is best characterized by disease-specific survival versus overall survival, which underestimates the impact of treatment in patients with favorable tumor and nodal stage. Subgroup analysis of patients with organ confinement and nodal status identified additional prognostic variables within the more favorable patient categories not apparent in the entire population. The poor prognosis of patients with NOC and/or N+ tumors emphasized the importance of future randomized trials in which such stratification variables may be of value. PMID- 12074818 TI - Improving inter-observer agreement and certainty level in diagnosing and grading papillary urothelial neoplasms: usefulness of a Bayesian belief network. AB - BACKGROUND AND OBJECTIVE: A Bayesian belief network (BBN), as diagnostic decision support system, enables the processing of our knowledge of histopathology expressed in descriptive terms, words and concepts. The aim of this study was to evaluate the contribution of a BBN in the improvement of inter-observer agreement and certainty level in the diagnosis and grading of papillary urothelial neoplasms. MATERIALS: Inter-observer agreement and certainty level were investigated on 40 cases of non-invasive papillary urothelial neoplasms subdivided according to the WHO 1973 classification. There were 10 urothelial papillomas (UPs), 10 grade 1 papillary carcinomas (G1), 10 grade 2 papillary carcinomas (G2) and 10 grade 3 papillary carcinomas (G3). Five consecutive sessions were held with three observers (RMa, PC and MSt). Sessions A, B and D were based on the morphological evaluation of the specimens with a conventional light microscope only. In sessions C and E, a BBN was used in addition to the microscope. The BBN output was represented by four belief values for four possible diagnostic outcomes. These values ranged from 0.0 to 1.0, with the sum of the belief values being 1.0. Concerning the certainty level, a two-tier system of assessment was adopted in sessions A, B and D: certain versus less certain. In sessions C and E, a belief value equal to or greater than 0.65 was considered as equivalent to "certain". RESULTS: In session A, an all-encompassing or synthetic approach to decision-making was adopted. Agreement with the gold standard was seen in 60% (RMa), 55% (PC) and 65% (MSt) of cases, respectively. The level of subjective confidence was "certain" in 35%, 40% and 35% of cases, respectively. Better agreement-70% (RMa), 68% (PC) and 72% (MSt) of cases-was present in session B where an analytical approach based on the evaluation of a series of morphological features was used. The level of subjective confidence was "certain" in 45%, 50% and 55% of cases, respectively. In session C, where a BBN was utilised, a further increase in degree of agreement with the gold standard was observed, e.g. 85% (RMa), 80% (PC) and 86% (MSt) of cases, respectively. Levels of certainty or belief values were high. Decrease in both the level of agreement 60% (RMa), 62% (PC) and 65% (MSt) of cases-and certainty was seen in session D where the observers were left free to evaluate the cases morphologically without the constrain of either a synthetic or analytical approach. In session E, where the BBN was used again, the percentage of cases in agreement with the gold standard increased to 83% (RMa), 81% (PC) and 84% (MSt), respectively. Increase in certainty or belief was also seen. The difference of the results obtained in the sessions A, B and D with those seen in the BBN-based sessions (C and E) is statistically significant. CONCLUSIONS: Conventional morphological evaluation of papillary urothelial neoplasms is affected by inter-observer variability and, in many instances, by diagnostic uncertainty. The greatest difficulties are found with G1 and G2 cases. Improvement in inter-observer agreement and certainty level can be achieved with a BBN. PMID- 12074819 TI - Pretreatment with interleukin-2 modulates peri-operative immuno-dysfunction in patients with renal cell carcinoma. AB - OBJECTIVE: Complex peri-operative immuno-dysfunction occurs in patients with renal cell carcinoma undergoing nephrectomy. Here, the effect of pretreatment with interleukin-2 (IL-2) is addressed. METHODS: Of 63 patients who underwent tumor nephrectomy, 26 patients received 4 doses of 10 Mio IE/m(2) IL-2 b.d. s.c. (i.e. a total of 40 Mio IE/m(2)) a week before operation, 37 did not. Parameters of cellular and humoral immunity (differential blood count, T-cell markers CD2, CD3, CD4, and CD8, B-cell markers CD19 and CD20, monocyte markers CD13 and CD14, NK (natural killer)-cell marker CD16, activation markers CD25, CD26, CD69, and HLA-DR, and cytokines IL-1-receptor antagonist (IL-1RA), IL-2, soluble IL-2 receptor (sIL-2R), IL-6, IL-10, and TGFbeta) were measured in venous blood. Blood was drawn before IL-2, 1 day before and immediately after the operation, and on the 1st, 3rd, 5th, and 10th postoperative day. RESULTS: All patients showed postoperatively elevated leukocyte and granulocyte counts, and elevated serum levels of cytokines IL-6 and IL-10. T-cell and activation markers were decreased. However, all these alterations were less accentuated in patients who had been pretreated with IL-2. Monocyte counts and IL-2 and TGFbeta levels were decreased, but IL-1RA and sIL-2R levels were elevated in pretreated patients. IL-2 related toxicity was WHO grades I-II in all patients, grade III in one patient. The anesthetic regimen had no measurable effect. IL-6 concentrations were higher in renal venous than in venous pool blood, indicating IL-6 production in the tumor in vivo. CONCLUSIONS: Pretreatment with IL-2 modulates peri-operative immuno dysfunction in patients undergoing tumor nephrectomy. This affects in particular T-cell-mediated immunity and levels of cytokines IL-10 and IL-6. The IL-2 administration scheme used here was followed by distinct counter-regulation including monocytes, IL-2, sIL-2R, IL-1RA and TGFbeta. PMID- 12074820 TI - Burch colposuspension and tension-free vaginal tape in the management of stress urinary incontinence in women. AB - OBJECTIVE: Objective of the study was to compare the efficacy and the complications of tension-free vaginal tape (TVT) and Burch colposuspension in the treatment of female genuine stress incontinence (GSI). METHODS: In this controlled, prospective, randomized study, participated 35 patients who underwent Burch colposuspension and 36 patients that underwent TVT procedure. Patients with prolapse more than first degree, previous surgical treatment of stress urinary incontinence (SUI) and detrusor instability were excluded from the study. RESULTS: The operative time for TVT was significantly shorter compared to BC. The severity and duration of postoperative pain for TVT was significantly less compared to BC. The necessary time for return to normal activity was 10 days for TVT and 21 days for BC. The cure rate after 24 months of follow-up was as follows: TVT: 84% and BC: 86%, while the improvement was 7% for TVT and 6% for BC. CONCLUSIONS: TVT and Burch colposuspension are equally effective in the management of female GSI at two years follow-up. TVT procedure requires much less operative time, has much shorter hospitalization time, with significantly less postoperative pain and faster return to normal daily activities than Burch colposuspension. PMID- 12074821 TI - Long term results of percutaneous treatment of caliceal diverticular calculi. AB - PURPOSE: We reviewed the long term outcome of percutaneous caliceal diverticular stone extraction. The objective was to define factors of treatment failure and recurrence. MATERIAL AND METHODS: Percutaneous caliceal diverticular stone extraction was performed in 24 women and 7 men (age range 21-69 years old). Diverticula were located throughout the kidney including the upper, middle and lower calices. In all cases stones were removed using a direct approach and diverticular neck was incised or dilated. Fulguration of the diverticular walls was performed in 6 cases. Trans-diverticular drainage was maintained from 3 to 5 days (mean 3.4 days) until a nephrostomogram demonstrated no extravasation. Patients were evaluated at 3 months and yearly thereafter. Success criteria were: no symptoms, no stone recurrence, no diverticulum left as assessed by IVP. Evaluation at 1 year minimum is presented. RESULTS: The average operative time and hospital stay were 103 min (range 90-130) and 3 days (range 4-9 days), respectively. No major complications and no mortality were observed. At 1 year stone free rate was 84% and diverticulum obliteration was obtained in 68% of patients. Overall 88% of patients were asymptomatic at average followup 24.6 months (range from 18 to 96 months). In all cases morphological or symptomatic failures were related to the quality of immediate result at surgery which relates to the location of the diverticulum. CONCLUSION: The percutaneous management of diverticular caliceal stones is a well standardized technique, however, surgical alternatives must be discussed any time the percutaneous approach seems difficult because high failure rates are expected in these cases. PMID- 12074822 TI - The molecular basis of genetic lipodystrophies. AB - OBJECTIVES: Hyperinsulinemia is often associated with a cluster of metabolic abnormalities, which usually presents before the onset of frank diabetes. Lipodystrophy syndromes are frequently associated with hyperinsulinemia and may act as models for insulin resistance. Lipodystrophy is characterized in broad terms by loss of subcutaneous adipose tissue. Despite heterogeneous causes, which include both genetic and acquired forms, lipodystrophy syndromes have similar metabolic attributes, including insulin resistance, hyperlipidemia and diabetes. RESULTS: Recently, the molecular basis of two genetic forms of lipodystrophy, namely Dunnigan-type familial partial lipodystrophy (FPLD; MIM 151660) and Berardinelli-Seip complete lipodystrophy (BSCL; MIM 269700) have been reported. There is evidence for genetic heterogeneity for both types of lipodystrophy. In addition, murine models of lipodystrophy have provided key insights into alterations of metabolic pathways in lipodystrophy. CONCLUSIONS: Delineation of the human molecular genetic basis of two distinct forms of inherited lipodystrophy may have relevance for the common insulin resistance syndrome and for acquired lipodystrophy syndromes. PMID- 12074823 TI - Regarding "the toxicity of Callilepis laureola, a South African traditional herbal medicine.". PMID- 12074825 TI - Porous graphitic carbon shows promise for the rapid screening partial DPD deficiency in lymphocyte dihydropyrimidine dehydrogenase in Chinese, Indian and Malay in Singapore by using semi-automated HPLC-radioassay. AB - OBJECTIVE: Dihydropyrimidine dehydrogenase (DPD) catalyzes the degradation of thymine, uracil, and the chemotherapeutic drug 5-Fluorouracil. In general reverse phase high pressure liquid chromatography is the standard method for separating 5 [2-(14)C]Fluorouracil and 5-[2-(14)C]Fluoro-5,6-dihydrouracil. However, the use of 100% aqueous solution (as HPLC mobile phase) may collapse the C-18 bonded phase and result in a retention time shift. The aim of this study is to develop a rapid, reproducible, sensitive method for screening partial DPD deficiency in healthy volunteers. DESIGN AND METHODS: The activity of DPD was measured using 5 [2- (14)C]Fluorouracil (5-[2-(14)C]FUra) followed by separation of substrate and product 5-[2-(14)C]FUraH(2) with a 15 x 4.6 mm I.D., 5 microm particle size (d(p)) porous graphitic carbon (PGC) column (Hypercarb(R)) and HPLC with online detection of the radioactivity. This was standardized using the protein concentration of the cytosol (NanoOrange(R) Protein Quantitation). RESULTS: Complete baseline separation of 5-[2-(14)C]Fluorouracil (5-[2-(14)C]FUra) and 5 [2-(14)C]Fluoro-5,6-dihydrouracil (5-[2-(14)C]FUraH(2)) was achieved using a porous graphitic carbon (PGC) column. The detection limit for 5-[2-(14)C]FUraH(2) was 0.4 pmol. CONCLUSIONS: By using linear gradient separation (0.1% Trifluoroacetic acid [TFA] in water to 100% Methanol) protocols in concert with PGC columns (Hypercarb(R)), we have demonstrated that a PGC column has a distinct advantage over C-18 reverse phase columns in terms of column stability (pH 1-14). This method provides an improvement on the specific assay for DPD enzyme activity. It is rapid, reproducible and sensitive and can be used for routine screening for healthy and cancer patients for partial and profound DPD deficiency before treatment with 5- FUra. PMID- 12074826 TI - Development of an automated immunoassay for advanced glycosylation end products in human serum. AB - OBJECTIVE: Nonenzymatic reaction of protein and carbohydrate produce a series of brown fluorescent advanced glycosylation end products (AGEs). However, a convenient and rapid assay for serum AGEs level is currently unavailable. METHODS: We raised AGEs-specific polyclonal antibodies, which were used to develop a fully automated, noncompetitive, homogeneous, light-scattering immunoassay for serum AGEs. RESULTS: The assay requires a sample volume of 2 microL and takes a reaction time of 2 min. The coefficient of variation range from 1.8 to 6.1%, and the mean recovery rate was 98.6%. Comparative analysis shows moderate correlation with competitive ELISA (r = 0.8209, n = 52). The mean +/- SD concentration of AGEs in young and in older healthy subjects were 4.6 +/- 1.5 (n = 39) and 4.9 +/- 1.4 (n = 40), respectively. The level of AGEs was significantly higher in serum from patients with type II DM 7.8 +/- 4.8 (n = 89) than that from the normal subjects (p < 0.05). CONCLUSIONS: The automatic immunoassay for AGEs is appropriate for clinical use. PMID- 12074827 TI - Paraoxonase Gln-Arg(192) and Leu-Met(55) gene polymorphisms and enzyme activity in a population with a low rate of coronary heart disease. AB - OBJECTIVES: To assess whether paraoxonase (PON1) polymorphisms at positions 55 and 192 and/or their phenotypic expressions influence the risk of myocardial infarction (MI) in Spanish population. DESIGN AND METHODS: Two hundred and fifteen male survivors of a MI and their age-matched controls were included in the study. Lipids, apolipoproteins (apo) A-I and B, PON1 activity on paraoxon and phenylacetate and PON1 polymorphisms were determined. RESULTS: Genotype distribution was similar in patients and controls. Enzyme activities were lower in patients, but multiple logistic regression analysis did not show any independent association with a higher risk of MI. CONCLUSION: None of the PON1 polymorphisms or their corresponding measured activities are independent risk factors for MI in our population. PMID- 12074828 TI - Relation between butyrylcholinesterase K variant, paraoxonase 1 (PON1) Q and R and apolipoprotein E epsilon 4 genes in early-onset coronary artery disease. AB - OBJECTIVES: The common K variant of butyrylcholinesterase (BChE-K), an enzyme which metabolizes acetylcholine and organophosphates, has been associated with Alzheimer's disease, especially in the presence of the apolipoprotein E epsilon 4 allele (APOE-epsilon 4). Although APOE-epsilon 4 has been associated with the development of coronary artery disease (CAD), an association between the BChE-K variant and CAD has not been explored. Paraoxonase 1 (PON1), located within HDL, is an enzyme which also metabolizes organophosphates and may be antiatherogenic. The R192 variant of PON1 (PON1-R) has been associated with CAD. DESIGN AND METHODS: To determine whether BChE-K is also associated with premature CAD, we examined the frequency of BChE-K among patients with early-onset CAD (n = 150; < 50 yr) vs. late-onset CAD (n = 150; > 65 yr) by molecular analysis. We also examined the frequency of the PON1-R allele in both groups, and explored whether there was synergism between BChE-K and APOE-epsilon 4, BChE-K and PON1-R or PON1 R and APOE-epsilon 4. RESULTS: The frequency of the BChE-K allele tended to be greater among early-onset CAD patients compared to late-onset CAD patients (41.3% vs. 31.3%; p = 0.07), but without any significant difference between males and females. There was no difference in the prevalence of the PON1-R allele between those with early- or late-onset CAD (46.0% vs. 52.7%; p = 0.25). Twenty-two patients with early-onset CAD had both the BChE-K plus APOE-epsilon 4 alleles (14.7%) compared to 11 late-onset CAD patients (7.3%) (p = 0.04). There was no such association between BChE-K and PON1-R, nor PON1-R and APOE-epsilon 4. CONCLUSIONS: Our study suggests that there is a minor association between BChE-K and early-onset CAD, especially in the presence of the APOE-epsilon 4 allele. PMID- 12074829 TI - LDL and HDL subclass distribution in patients with end-stage renal diseases. AB - OBJECTIVES: To examine the alterations in LDL and HDL subclass distribution in ESRD patients compared with a control group and to investigate the relationship of LDL particle size to the other plasma lipoproteins levels. DESIGN AND METHODS: Plasma lipids, LDL and HDL subclasses were determined in 63 hemodialysis patients (HD), 42 predialysis patients and 345 control subjects. Lipoprotein subclasses were separated by polyacrylamide 3 to 31% gradient gel electrophoresis. RESULTS: In predialysis group, 88% subjects had small LDL particles compared with 58.5% of hemodialysis patients and 16.5% of control subjects. Mean LDL size particle diameter was significantly smaller in HD and predialysis patients in comparison with controls (p < 0,0005, p < 0,0001; respectively). Significant inverse correlation between LDL particle size and triglyceride level was observed for both patient groups. Decreased levels of the largest HDL2b subclass was found in both predialysis (16.5%) and in HD patients (30%) as compared with controls (50%), and increased levels of the small HDL3a subclass was found only in predialysis group (21%) in comparison with controls (4.5%). CONCLUSIONS: Alterations in LDL and HDL subclass distribution toward smaller particles is the main lipid abnormality associated with atherogensis found in ESRD. ESRD is associated with reduced levels of HDL2b subclass and increased levels of HDL3c subclass, which occurs in coronary artery disease (CAD) as well. PMID- 12074830 TI - The evaluation of autoantibodies against oxidatively modified low-density lipoprotein (LDL), susceptibility of LDL to oxidation, serum lipids and lipid hydroperoxide levels, total antioxidant status, antioxidant enzyme activities, and endothelial dysfunction in patients with Behcet's disease. AB - OBJECTIVES: Behcet's disease is a multisystem disorder characterized by a chronic inflammation including acute attacks and remission periods. Decreased enzyme activity of the antioxidant system and increased levels of free radicals may have important roles in the damage of tissues observed in the disease period. In addition, the atherogenic tendency of serum lipid, lipoproteins, lipid peroxidation levels and endothelial dysfunction accompany the above mentioned findings. As a consequence of these events, different degrees of low density lipoprotein (LDL) oxidation occur in vivo, and then autoantibodies against oxidized-LDL(AuAb-oxLDL) are produced. DESIGN AND METHODS: Lipids, lipoproteins, lipid hydroperoxide, AuAb-oxLDL, total antioxidant status (TAS), serum-soluble intercellular adhesion molecule-1 (sICAM-1), plasminogen activator inhibitor 1 (PAI-1) levels in serum, the activities of antioxidant enzymes including glutathione peroxidase (GSH-Px), glutathione reductase (GR), superoxide dismutase (SOD), and catalase (CAT) in erythrocytes and plasma, were determined in 25 patients with Behcet's disease and in 25 healthy volunteers. Also, susceptibility to copper-induced in vitro oxidation of LDL by using lag time, a measure of resistance to oxidation, oxidation rate and extent of oxidation, a measure of diene production in both groups, was studied. RESULTS: It was observed that lipid hydroperoxide and AuAb-oxLDL levels in patients with Behcet's disease were significantly higher, but erythrocyte SOD, CAT, plasma GSH-Px activities, and TAS were significantly lower than those in healthy subjects. Susceptibility of LDL to oxidation in the patients was found to be increased. Total cholesterol, LDL-C and apo B levels and acute phase reactants were significantly higher, but HDL-C and apo AI levels were significantly lower, in patients when compared to healthy subjects. The levels of AuAb-oxLDL in patients were found to correlate with TAS, total cholesterol, LDL-C, lipid hydroperoxide and erythrocyte SOD activities (r = -0.62, p < 0.01; r = 0.64, p < 0.01; r = 0.55, p < 0.01; r = 0.81, p < 0.01; r = 0.63, p < 0.01, respectively). In addition, lipid hydroperoxide levels were found to correlate with total cholesterol, LDL-C and erythrocyte SOD activities (r = 0.45, p < 0.05; r = 0.45, p < 0.05; r = -0.46, p < 0.05, respectively). PAI-1 and sICAM-1 were found to be increased in the patients and correlated with AuAb-oxLDL and lipid hydroperoxide levels (r = 0.56, p < 0.01; r = 0.67, p < 0.01 and r = 0.59, p < 0.01; r = 0.61, p < 0.01, respectively). CONCLUSIONS: It was concluded that the observed increase of lipid, lipoproteins, lipid hydroperoxide, susceptibility of LDL to oxidation, autoantibodies against ox-LDL levels and decrease of antioxidant enzyme activities and total antioxidant status and increased secretion of endothelial derivated peptides including sICAM and PAI-1, and their interactions may indicate that there is a tendency to atherothrombotic events in patients with Behcet's disease. PMID- 12074831 TI - Decreased concentration of human kallikrein 6 in brain extracts of Alzheimer's disease patients. AB - BACKGROUND: The human kallikrein 6 gene (KLK6) encodes for a secreted serine protease, hK6, which is highly expressed in brain. Previous reports have associated hK6 with the pathogenesis of Alzheimer's disease. Our objective was to develop a highly sensitive immunoassay for hK6 and use it to examine the levels of hK6 in brain tissue extracts from Alzheimer's disease patients and control subjects. METHODS: We developed antibodies against hK6 and constructed a 'sandwich' type immunoassay. We then assessed levels of hK6 in brain extracts from normal individuals and patients with Alzheimer's disease. RESULTS: The hK6 assay was developed using a combination of two antibodies (a mouse monoclonal and a rabbit polyclonal). Purified recombinant hK6 was used as a calibrator. The detection limit of the assay was 0.05 microg/L. The intra and inter-assay coefficient of variation was less than 6.5%. We found no detectable cross reactivity by the homologous proteins hK2, hK3, hK8, hK10, hK11, hK13 and hK14. The hK6 concentration in human brain tissue extracts from healthy (n = 24) and Alzheimer's patients (n = 55) were 10.1 +/- 1.0 and 3.39 +/- 0.26 mcirog/g of total protein (mean +/- SE), respectively (p < 0.001). Similar differences were seen when the tissues were stratified by brain region (occipital, parietal, frontal and temporal cortex). CONCLUSIONS: We conclude that the newly developed hK6 immunoassay is suitable for quantification of hK6 protein in biologic fluids and tissue extracts. The brain of Alzheimer's disease patients contains significantly less hK6 than the brain of nonaffected individuals. The possible connection of hK6 with the pathogenesis of Alzheimer's disease merits further investigation. PMID- 12074832 TI - Changes of plasma free choline and choline-containing compounds' concentrations and choline loss during hemodialysis in ESRD patients. AB - OBJECTIVES: This study was undertaken to determine the changes in plasma free choline and choline-containing compounds in end stage renal disease (ESRD) and to determine if they were lost into the dialysate during hemodialysis. DESIGN AND METHODS: Plasma and dialysate free choline, phosphocholine and phospholipid-, phosphatidylcholine-, sphingomyelin-bound choline were measured before, during and after hemodialysis. RESULTS: Plasma free and bound choline concentrations (mean +/- standard error of the mean) were 12.9 +/- 0.6 and 2697 +/- 57 microM or 37.3 +/- 0.9 and 2792 +/- 98 microM in controls or in ESRD patients, respectively. Free choline concentrations were correlated (r = 0.598; p < 0.001) with the time the patients were subjected to hemodialysis. Plasma free choline and phosphocholine concentrations are decreased by a total of -8.1 +/- 0.6 micromol/L and -88 +/- 8 micromol/L, respectively; phospholipid-, phosphatidylcholine- and sphingomyelin-bound choline are increased, during hemodialysis. Patients lost about 350 micromoles of choline into the dialysate during hemodialysis. CONCLUSION: Plasma free choline concentrations are elevated in ESRD, and a considerable amount of choline is lost into the hemodialysate. PMID- 12074833 TI - Antioxidant potential of propylthiouracil in patients with psoriasis. AB - OBJECTIVE: There is growing evidence supporting the reactive oxygen species (ROS) in the pathogenesis of psoriasis. Propylthiouracil(PTU), an antithyroid drug, has been shown to have beneficial effects on psoriasis. The aim of this study was to investigate both disturbances in oxidant/antioxidant system in psoriasis and whether PTU, shown to have immunomodulatory effects and antioxidant potential, has effects on oxidant/antioxidant system and clinical improvement in psoriatics. DESIGN AND METHODS: Malondialdehyde (MDA), end product of lipid peroxidation, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and antioxidant enzymes were measured in plasma, erythrocytes and skin biopsies of psoriatics who were resistant to conventional therapy before and after 8 weeks of oral treatment with PTU (300 mg/day) or PTU/thyroxine (25 microg/day- to prevent possible hypothyroidism). The same parameters were also studied in healthy controls. Psoriasis Area and Severity Index (PASI) scores were used to evaluate the severity of the disease, and routine analyses and thyroid function tests were measured during the study. RESULTS: Increased baseline MDA in all samples were found to be lower. In addition baseline SOD and GSH-Px in skin and erythrocytes were also lower. The increased plasma SOD levels in skin and erythrocytes of the study groups was found to be higher and lower,respectively in all patients after the treatment. No tissue parameters or erythrocyte GSH-Px were different from control levels at the end of the study. Significant clinical improvement and decreased PASI scores were observed in all patients. Post treatment TSH levels were higher in all patients, but these levels were within the reference range and none had clinical hypothyroidism. CONCLUSION: These findings may provide some evidence for a potential role of increased lipid peroxidation and decreased antioxidant activity in psoriasis. PTU may be considered as treatment model in psoriasis, in particular for resistant cases, because of its antioxidant potential, and also antiproliferative and immunomodulatory effects. PMID- 12074834 TI - The effect of smoking on bone metabolism: maternal and cord blood bone marker levels. AB - BACKGROUND: To clarify the effect of smoking on bone metabolism in the fetus, we measured osteocalcin (OC), bone isoenzyme of alkaline phosphatase (BALP), procollagen type 1 C-terminal propeptide (PICP) in maternal serum and umbilical cord blood. METHODS: 15 active smoker, 14 passive smoker, 15 nonsmoker women and their newborn were included in this study. OC, BALP, PICP were determined by enzyme immunoassay. RESULTS: Of the bone markers tested only OC was different in the serum of the three groups of women. Infants of smoker women have significantly lower umbilical cord blood OC levels than those of infants from both passive smoker and nonsmoker women.(25.6 +/- 6.6, 35.8 +/- 10.4, 37.2 +/- 16.1 ng/mL respectively, p < 0.05). Infants of smoker women have significantly lower umbilical cord blood BALP levels than those of infants from nonsmoker women. (46 +/- 12, 57 +/- 15 U/L p < 0.05). All bone markers except total ALP were significantly higher in umbilical cord blood as compared to maternal blood levels (p < 0.001 for all). CONCLUSION: High umbilical cord blood bone marker levels may reflect the altered bone metabolism of fetus. Moreover, chronic hypoxia due to smoking may cause the suppression of bone matrix synthesis or placental synthesis as reflected by low OC and BALP levels in umbilical cord blood of infants from smoker women. PMID- 12074835 TI - Comparison between the Cobas Integra immunoassay and high-performance liquid chromatography for therapeutic monitoring of carbamazepine. AB - OBJECTIVES: Carbamazepine (CBZ) is metabolized by the cytochrome P450 into carbamazepine-10,11-epoxide (CBZepox), a metabolite with similar pharmacological activity to the parent drug. Recently it has been indicated that most current immunoassays for the determination of CBZ are unable to quantitatively measure its active epoxide. An evaluation of the Cobas Integra immunoassay for the determination of CBZ was carried out, and the results are compared with those obtained for CBZ+CBZepox using high-performance liquid chromatography (HPLC) in patients on monotherapy and polytherapy. DESIGN AND METHODS: Steady-state serum trough concentrations of CBZ were determined in 119 epileptic patients using the Cobas Integra immunoassay and HPLC. In 91 cases CBZ was administered in monotherapy and in 28 cases in polytherapy with other anticonvulsant drugs. RESULTS: The study of within- and between-run imprecisiom for the Cobas Integra immunoassay led to clinically acceptable coefficients of variation. A high correlation was found between the concentrations of CBZ obtained using the immunoassay and HPLC (r = 0.981, p < 0.001). In both the group of patients on monotherapy and those on polytherapy, the levels of CBZepox were greater than the clinically acceptable error for CBZ; consequently, there is a clinically significant difference between the total of CBZ+CBZepox concentrations (HPLC) and the concentrations of CBZ (immunoassay). In the group of patients under monotherapy, a high correlation coefficient was obtained between the levels of CBZ and CBZ+CBZepox (r = 0.975, p < 0.001) with an standard error of the estimate similar to the clinically acceptable value. CONCLUSIONS: For the patients on monotherapy, it is possible to make a clinically valid estimation of CBZ+CBZepox from the concentration of CBZ obtained by means the immunoassay. In patients on polytherapy, the analytical determination of CBZepox could be of interest in cases where CBZ+CBZepox would be higher than the critical level of CBZ. PMID- 12074836 TI - Effects of ablation of N- and R-type Ca(2+) channels on pain transmission. AB - Recently several mutant mouse lines lacking neuronal voltage-dependent Ca(2+) channels (VDCCs) have been established by the use of gene targeting in embryonic stem cells. Pain-related behaviors in Ca(v)2.2 (alpha(1B)) and Ca(v)2.3 (alpha(1E)) knockout mice were studied to gain further insight into the mechanism of pain transmission, where VDCCs are thought to play important roles. We review here the data from these recent studies. Ca(v)2.3-/- mice showed normal responses to acute painful stimuli, and reduced responses to the somatic inflammatory pain stimuli. Ca(v)2.3+/- mice exhibited reduced symptoms of visceral inflammatory pain. Ca(v)2.3-/- mice showed abnormal behavior related to the descending antinociceptive mechanism activated by the intraperitoneal injection of acetic acid. Ca(v)2.2-/- mice showed variable acute nociceptive responses depending on the mutant lines. However, all the lines of Ca(v)2.2-/- mice exhibited reduced responses in the phase 2 of the formalin test, suggesting a suppression of inflammatory pain. Furthermore Ca(v)2.2-/- mice showed markedly reduced neuropathic pain symptoms after spinal nerve ligation. Impaired antinociception, similar to that seen in the Ca(v)2.3-/- mice, was also observed in the Ca(v)2.2-/ mice. Therefore, it is suggested that these mutant mice could provide novel models to delineate the nociceptive and antinociceptive mechanisms. PMID- 12074837 TI - Neuronal activity representing visuospatial mnemonic processes associated with target selection in the monkey dorsolateral prefrontal cortex. AB - To investigate how visuospatial mnemonic and target selection processes are represented in the dorsolateral prefrontal cortex (PFC), we studied neuronal attributes of the dorsolateral PFC while monkeys were performing oculomotor delayed visual search (ODVS) and oculomotor delayed-response (ODR) tasks. In the ODVS task, the subject made a memory-guided saccade to a remembered target location that had been presented along with distractors before a delay period; in the ODR task, the target was presented without any distractors. A total of 252 neurons in the dorsolateral PFC showed directional delay-period activity and were divided into two groups; neurons that showed directional delay-period activity predominantly in the ODVS task (n=112), and those that showed such activity similarly in both the ODVS and ODR tasks (n=140). These neuronal groups shared similar temporal properties (i.e. onset latency, peak time of delay-period activity) and spatial tuning. Our findings suggest that the dorsolateral PFC contains a particular visuospatial memory system for information selected by target selection (selective attention), and this attention-memory system (or 'memory system for special use') appears to be represented in the dorsolateral PFC, in parallel with a more 'general' memory system that is not specifically associated with target selection. PMID- 12074838 TI - Different patterns of dipole source localization in gelastic seizure with or without a sense of mirth. AB - Dipole source localization corresponding to interictal spikes were estimated using EEG dipole tracing with a realistic three-shell head model in three patients with cryptogenic gelastic epilepsy. The dipole sources in two patients, whose gelastic seizures were accompanied by a subjective feeling of mirth, were estimated in the right or left medio-basal temporal regions. In the other patient, with gelastic seizures without a sense of mirth, the dipole sources were localized in the right frontal region corresponding to the anterior cingulate. The results suggest that the neural activities in hippocampal regions are involved with the generation of gelastic seizures with a sense of mirth and those in the cingulate might be associated with the motor act of laughter. PMID- 12074839 TI - Involvement of nitric oxide in glucose toxicity on differentiated PC12 cells: prevention of glucose toxicity by tetrahydrobiopterin, a cofactor for nitric oxide synthase. AB - Effects of high concentrations of glucose on cell survival of differentiated PC12 cells were examined. Seven day-culture with D-glucose (9.0-27.0 mg/ml as 2-6-fold of the optimal level) induced cell death in a dose-related manner but 3-day culture with high concentrations of glucose had no effect on cell viability. L glucose had no effect on viability of PC12 cells, suggesting that D-glucose toxicity was independent of its osmolarity effect. Seven-day culture with D glucose (13.5 mg/ml as 3-fold of the optimal level) increased nitric oxide metabolites (NOx) in the culture medium. Glucose-induced increase in NOx was eliminated by 0.1 mM L-nitro-arginine methylester (L-NAME), a nitric oxide synthase (NOS) inhibitor. Intracellular Ca(2+) concentration was increased by D glucose in a dose-related manner, suggesting that D-glucose activated NOS by increasing intracellular Ca(2+) concentration in PC12 cells. Glucose-induced cell death was blunted by 0.1 mM L-NAME, showing that nitric oxide (NO) was involved in the glucose toxicity to PC12 cells. Tetrahydrobiopterin (BH(4)), a cofactor for NOS, attenuated both glucose-induced cell death and NOx production at 1 microM but not at 10 microM. The effects of BH(4) on glucose-induced cell death and NOx production were not mimicked by reducing agents such as ascorbate and cysteine. These results taken together suggest that high concentrations of glucose induced cell death via NO production and that low concentration of BH(4) had a protective effect against glucose neurotoxicity in differentiated PC12 cells. PMID- 12074840 TI - Abeta-degrading endopeptidase, neprilysin, in mouse brain: synaptic and axonal localization inversely correlating with Abeta pathology. AB - Metabolism of amyloid-beta peptide (Abeta) is closely associated with the pathology and etiology of Alzheimer's disease (AD). Since neprilysin is the only rate-limiting catabolic peptidase proven by reverse genetics to participate in Abeta metabolism in vivo, we performed detailed immunohistochemical analysis of neprilysin in mouse brain using neprilysin-deficient mice as a negative control. The aim was to assess, at both the cellular and subcellular levels, where Abeta undergoes neprilysin-dependent degradation in the brain and how neprilysin localization relates to Abeta pathology in amyloid precursor protein (APP) transgenic mice. In hippocampus, neprilysin was present in the stratum pyramidale and stratum lacunosum-moleculare of the CA1-3 fields and the molecular layer of the dentate gyrus. Confocal double immunofluorescence analyses revealed the subcellular localization of neprilysin along axons and at synapses. This observation suggests that after synthesis in the soma, neprilysin, a type II membrane-associated protein, is axonally transported to the terminals, where Abeta degradation is likely to take place. Among various cell types, GABAergic and metabotropic glutamate 2/3 receptor-positive neurons but not catecholaminergic or cholinergic neurons, expressed neprilysin in hippocampus and neocortex, implying the presence of a cell type-specific mechanism that regulates neprilysin gene expression. As expected, Abeta deposition correlated inversely with neprilysin expression in TgCRND8 APP-transgenic mice. These observations not only support the notion that neprilysin functions as a major Abeta-degrading enzyme in the brain but also suggest that down-regulation of neprilysin activity, which may be caused by aging, is likely to elevate local concentrations of Abeta at and around neuronal synapses. PMID- 12074841 TI - Cortical diffusible factors increase MAP-2 immunoreactive neuronal population in thalamic cultures. AB - Previous experiments have established that grafts of embryonic day (E) 16 frontal cortex placed into the occipital cortex of postnatal day (P) 0-P1 rats selectively attract axons from the ventrolateral and ventromedial (VL/VM) thalamic nuclei (Frappe et al., Exp. Neurol. 169 (2001) 264). The present study was therefore undertaken to identify any possible maturation-promoting activity of the cortex on VL/VM thalamic cells. In a first step, a primary culture of VL/VM thalamic cells taken from P0-P1 rats was developed. Neurons, glial cells and a few immature, nestin immunoreactive cells were identified in the culture. In a second step, VL/VM thalamic cells that had been maintained in vitro for 4-5 days were cultured for 7 additional days in isolation (control condition) or with an E16 or P5 explant of frontal or occipital cortex placed on a microporous membrane. In control conditions, the total cell population and the percentage of MAP-2 immunoreactive neurons were not modified with time. In contrast, the percentage of MAP-2 immunoreactive neurons was increased in E16 cortex co cultures whereas the total cell population was unchanged and the proliferative activity remained very low. Also, the mean number of neurites per neuron was increased but no effect was found on neuritic length. Similar effects on neuronal maturation were found with E16 frontal or occipital cortex explants, indicating a lack of areal specificity. P5 cortex also produced, but to a lesser extent, an increase in percentage of MAP-2 immunoreactive neurons. Further, P5 cortex had no effect on mean number of neurites per neuron but substantially promoted elongation of neuronal processes. We propose that in addition to their well established survival promoting effect, diffusible molecules released by embryonic and early postnatal cortex can promote in vitro the maturation of thalamic neurons. PMID- 12074842 TI - Brain-specific angiogenesis inhibitor 1 (BAI1) is expressed in human cerebral neuronal cells. AB - Brain-specific angiogenesis inhibitor 1 (BAI1) is a p53-target gene specifically expressed in the brain. We examined the distribution of the endogenous BAI1 protein in normal human brain tissue using a polyclonal antibody against the extracellular region of BAI1. Immunohistochemical study demonstrated that BAI1 was expressed in neuronal cells of the cerebral cortex but not in astrocytes. BAI1 protein was localized in the cellular cytoplasm and membrane. It was predominantly localized in the cellular membrane when expressed in cultured cells by means of gene transfection. BAI1 protein may play an important role in neuronal functions such as synapse formation and signal transduction. PMID- 12074843 TI - Combined argatroban and edaravone caused additive neuroprotection against 15 min of forebrain ischemia in gerbils. AB - We investigated whether or not a combination of the selective thrombin inhibitor, argatroban, and the free radical scavenger, edaravone (MCI-186), ameliorates postischemic hypoperfusion and decreases mortality after 15 min of forebrain ischemia in the gerbil. Argatroban or edaravone alone significantly increased postischemic cerebral blood flow and attenuated brain edema after reperfusion. However, only the combination increased the survival ratio (P<0.05 by Mantel-Cox) and protected the damage of neuronal cells. The present study indicates that anticoagulants and free radical scavengers reciprocally function to inhibit the progression of ischemic cell damage and that a combination of these types of drugs will help to improve the outcomes after cerebral ischemia. PMID- 12074844 TI - Delivery of antisense oligonucleotides to PC12 cells. AB - Optimal experimental conditions for the delivery of phosphodiester or phosphorothioate antisense oligonucleotides (P-ASO/S-ASO) to PC12 cells were determined. Fluorescently labeled P-ASO or S-ASO were transfected to PC12 cells and the uptake of antisense, free or entrapped in liposomes, was monitored by confocal and fluorescent microscopy. Efficient delivery of fluorescently labeled ASO with low rates of cell death was obtained when PC12 cells were transfected with liposomes in Opti-MEM medium supplemented with sera, compared with control experiments where nonliposomal ASO were transfected to PC12 cells in sera-free media. Compared with P-ASO, the application of S-ASO for antisense studies in PC12 cells is more suitable due to the lower concentration required for an efficient antisense uptake and its higher intracellular stability. PMID- 12074845 TI - Segmentation of fat and muscle from MR images of the thigh by a possibilistic clustering algorithm. AB - Physical training is proved to induce changes in physical capacity and body composition. We propose in this article a fast, unsupervised and fully three dimensional automatic method to extract muscle and fat volumes from magnetic resonance images of thighs in order to assess these changes. The technique relies on the use of a fuzzy clustering algorithm and post-processings to accurately process the body composition of thighs. Results are compared on 11 healthy voluntary elderly people with those provided on the same data by a validated method already published, and its reliability is assessed on repeated measures on three subjects. The two methods statistically agree when computing muscle and fat volumes, and clinical implications of this fully automatic method are important for medicine, physical conditioning, weight-loss programs and predictions of optimal body weight. PMID- 12074846 TI - A zero-inflated Poisson mixed model to analyze diagnosis related groups with majority of same-day hospital stays. AB - With increasing trend of same-day procedures and operations performed for hospital admissions, it is important to analyze those Diagnosis Related Groups (DRGs) consisting of mainly same-day separations. A zero-inflated Poisson (ZIP) mixed model is presented to identify health- and patient-related characteristics associated with length of stay (LOS) and to model variations in LOS within such DRGs. Random effects are introduced to account for inter-hospital variations and the dependence of clustered LOS observations via the generalized linear mixed models (GLMM) approach. Parameter estimation is achieved by maximizing an appropriate log-likelihood function using the EM algorithm to obtain approximate residual maximum likelihood (REML) estimates. An S-Plus macro is developed to provide a unified ZIP modeling approach. The determination of pertinent factors would benefit hospital administrators and clinicians to manage LOS and expenditures efficiently. PMID- 12074847 TI - Calculating adjusted R(2) measures for Poisson regression models. AB - In regression models not only the parameter estimates and significances of explanatory variables are of interest, but also the degree to which variation in the dependent variable can be explained by covariates. In recent publications, an R(2) measure based on deviance was recommended for Poisson regression models, one of the most frequently used modelling tools in epidemiological studies. However, when sample size is small relative to the number of covariates in the model, simple R(2) measures may be seriously inflated and may need to be adjusted according to the number of covariates in the model. We present a SAS-macro that calculates adjustments for the R(2) measures in Poisson regression models based on log-likelihood and on sums of squares. The proposed measures are applied to real data sets and their performance is discussed. PMID- 12074848 TI - Computer assisted measurement of airway gland secretions by the hillocks technique. AB - We describe a computer-based image acquisition and analysis system designed for the quantitation of airway submucosal gland fluid flux by the hillocks technique. This technique is based on the detection of the increase in surface area of hillocks formed above gland duct openings by secretions captured under a thin layer of tantalum powder. The advantages of the system are: (a) ability to detect and measure individual submucosal gland secretions, (b) computer assisted data acquisition and analysis, and (c) decreased propagated error in hillock volume measurements. Test results of a practical implementation of the system demonstrate a swine tracheal submucosal gland flux induced by the muscarinic agonist acetylcholine (50 microM) of 0.65 microl/min/cm(2). PMID- 12074849 TI - Tele-medical imaging conference system based on the Web. AB - In this paper, a medical imaging conference system is presented, which is carried out in the Web environment using the distributed object technique, CORBA. Independent of platforms and different developing languages, the CORBA-based medical imaging conference system is very powerful for system development, extension and maintenance. With this Web client/server, one could easily execute a medical imaging conference using Applets on the Web. The Java language, which is object-oriented and independent of platforms, has the advantage of free usage wherever the Web browser is. By using the proposed system, we envisage being able to open a tele-conference using medical images, e.g. CT, MRI, X-ray etc., easily and effectively among remote hospitals. PMID- 12074850 TI - A Bayesian approach to estimate evoked potentials. AB - Several approaches, based on different assumptions and with various degree of theoretical sophistication and implementation complexity, have been developed for improving the measurement of evoked potentials (EP) performed by conventional averaging (CA). In many of these methods, one of the major challenges is the exploitation of a priori knowledge. In this paper, we present a new method where the 2nd-order statistical information on the background EEG and on the unknown EP, necessary for the optimal filtering of each sweep in a Bayesian estimation framework, is, respectively, estimated from pre-stimulus data and obtained through a multiple integration of a white noise process model. The latter model is flexible (i.e. it can be employed for a large class of EP) and simple enough to be easily identifiable from the post-stimulus data thanks to a smoothing criterion. The mean EP is determined as the weighted average of the filtered sweeps, where each weight is inversely proportional to the expected value of the norm of the correspondent filter error, a quantity determinable thanks to the employment of the Bayesian approach. The performance of the new approach is shown on both simulated and real auditory EP. A signal-to-noise ratio enhancement is obtained that can allow the (possibly automatic) identification of peak latencies and amplitudes with less sweeps than those required by CA. For cochlear EP, the method also allows the audiology investigator to gather new and clinically important information. The possibility of handling single-sweep analysis with further development of the method is also addressed. PMID- 12074851 TI - Personal health care services through digital television. AB - Personal health care has obtained increasing importance in the field of health care as the populations' age in the industrialised countries and resources available for health care remain limited. Personal health care through digital television is an exiting possibility in the realisation of new types of services answering to this demand for increased personal action and responsibility in health care. The possibilities of digital television in health care are studied in the Health Care Television (HCTV) research project of the Digital Media Institute at Tampere University of Technology. In this paper personal health care services are studied mainly from the perspective of the interactive service infrastructure of digital television. Firstly we present the general infrastructure of digital television and the different interactive service types of digital television. The usage of these service types in personal health care applications is also discussed. Finally, a web-based application based on chronic atrial fibrillation and its test use is presented. The application is used as a research platform for personal health care applications in digital television. PMID- 12074852 TI - Commentary: dermoscopy. PMID- 12074853 TI - Impact of dermoscopy on the clinical management of pigmented skin lesions. PMID- 12074854 TI - Early detection of melanoma: the best strategy for a favorable prognosis. PMID- 12074855 TI - Clinical simulators of melanoma. PMID- 12074856 TI - Comparison between morphological parameters in pigmented skin lesion images acquired by means of epiluminescence surface microscopy and polarized-light videomicroscopy. PMID- 12074857 TI - Dermoscopic-pathologic correlation: an atlas of 15 cases. PMID- 12074858 TI - Pattern analysis: a two-step procedure for the dermoscopic diagnosis of melanoma. PMID- 12074859 TI - Dermoscopy: alternative melanocytic algorithms-the ABCD rule of dermatoscopy, Menzies scoring method, and 7-point checklist. PMID- 12074860 TI - Vascular patterns in skin tumors. PMID- 12074861 TI - Dermoscopic classification of Clark's nevi (atypical melanocytic nevi). PMID- 12074862 TI - Dermoscopic classification of Spitz/Reed nevi. PMID- 12074863 TI - Surface microscopy features of congenital nevi. PMID- 12074864 TI - Dermoscopy of pigmented basal cell carcinoma. PMID- 12074865 TI - Dermoscopic diagnosis of seborrheic keratosis. PMID- 12074866 TI - Dermoscopic diagnosis of vascular lesions. PMID- 12074867 TI - Dermatoscopy for facial pigmented skin lesions. PMID- 12074868 TI - Dermoscopy for acral pigmented skin lesions. PMID- 12074869 TI - Oral pigmented lesions. PMID- 12074870 TI - Pink lesions. PMID- 12074871 TI - Follow-up of melanocytic skin lesions with digital total-body photography and digital dermoscopy: a two-step method. PMID- 12074872 TI - Dermoscopy and preoperative evaluation of melanoma thickness. PMID- 12074873 TI - Digital dermoscopy analysis for automated diagnosis of pigmented skin lesions. PMID- 12074875 TI - Skin changes and the face. Catvat/Dubrovnik, Croatia--September 20-23, 2001. PMID- 12074876 TI - Ethnic hair and skin: what is the state of the science?. Chicago, Illinois- September 29-30, 2001. PMID- 12074877 TI - Ras family therapy: Rab, Rho and Ral talk to the exocyst. PMID- 12074884 TI - MAPK signaling specificity: it takes two to tango. AB - A surprisingly limited repertoire of core signaling pathways generates an enormous diversity of responses, often in a cell type-specific manner. Even within one cell, a single pathway might yield two different responses depending on the input signal. In budding yeast, a mitogen-activated protein kinase (MAPK) module seems to transmit both mating pheromone and invasive growth signals in the same yeast cell. We discuss recent insights into mechanisms of differential MAPK activation in this system. PMID- 12074885 TI - Crosstalk between cAMP and MAP kinase signaling in the regulation of cell proliferation. AB - Hormonal stimulation of cyclic adenosine monophosphate (cAMP) and the cAMP dependent protein kinase PKA regulates cell growth by multiple mechanisms. A hallmark of cAMP is its ability to stimulate cell growth in many cell types while inhibiting cell growth in others. In this review, the cell type-specific effects of cAMP on the mitogen-activated protein (MAP) kinase (also called extracellular signal-regulated kinase, or ERK) cascade and cell proliferation are examined. Two basic themes are discussed. First, the capacity of cAMP for either positive or negative regulation of the ERK cascade accounts for many of the cell type specific actions of cAMP on cell proliferation. Second, there are several specific mechanisms involved in the inhibition or activation of ERKs by cAMP. Emerging new data suggest that one of these mechanisms might involve the activation of the GTPase Rap1, which can activate or inhibit ERK signaling in a cell-specific manner. PMID- 12074886 TI - XMAP215: a key component of the dynamic microtubule cytoskeleton. AB - Microtubules are essential for various cellular processes including cell division and intracellular organization. Their function depends on their ability to rearrange their distribution at different times and places. Microtubules are dynamic polymers and their behaviour is described as dynamic instability. Rearrangement of the microtubule cytoskeleton is made possible by proteins that modulate the parameters of dynamic instability. Studies using Xenopus egg extracts led to identification of a microtubule-associated protein called XMAP215 as a major regulator of physiological microtubule dynamics. XMAP215 belongs to an evolutionarily conserved protein family present in organisms ranging from yeast to mammals. Together with members of the Kin I family of kinesins, XMAP215 and its orthologues form an essential circuit for generating dynamic microtubules in vivo. PMID- 12074887 TI - The low-density lipoprotein receptor gene family: a cellular Swiss army knife? AB - The low-density lipoprotein receptor gene family is an evolutionarily conserved group of cell-surface receptors produced by mammals and other organisms. Initially thought to be endocytic receptors that mediate the uptake of lipoproteins, recent findings have shown that these receptors have other roles in a range of cellular processes. Among other activities, members of this family act as signal transducers in neuronal migration processes, regulate synaptic plasticity or control vitamin homeostasis. Such multifunctionality is achieved by interaction with diverse cell-surface proteins including glycolipid-anchored receptors, G-protein-coupled receptors and ion channels. Here, we review the molecular interactions of this protein family with other cell-surface proteins that provide specificity and versatility - a versatility that may be reminiscent of a cellular Swiss army knife. PMID- 12074888 TI - Reverse genetics of essential genes in tissue-culture cells: 'dead cells talking'. AB - In the 'post-genomic' era, cDNA and genomic sequences are now available that encode huge numbers of proteins. Assigning functions to these proteins is a daunting task. Cell biologists have traditionally approached this problem by disrupting protein function with dominant-negative or structural mutants. Here, we describe several alternative approaches whereby cells or cell lines lacking particular gene products can be generated from genomic sequences for use in functional studies. These include gene targeting in mouse, human and chicken DT40 cells, and recent advances in double-stranded RNA-mediated interference (RNAi). PMID- 12074889 TI - Profile: Tom Rapoport interviewed by Rabiya S. Tuma. PMID- 12074892 TI - Functional expression of CXCR3 in cultured mouse and human astrocytes and microglia. AB - It has been established recently that inflammation of the CNS is accompanied by an expression of chemokines within the CNS. Several lines of evidence suggest that chemokines within the CNS initiate and orchestrate the infiltration of the inflamed brain by blood leukocytes. It is also known that endogenous cells of the CNS express functional chemokine receptors, raising the possibility that chemokines may be involved in intercellular signalling between brain cells during brain inflammation. It was shown recently that two chemokine ligands for CXCR3 are induced rapidly in damaged neurons. Little is known yet on the function of neuronal chemokine expression. In order to investigate whether neuronal chemokines contribute to endogenous signalling within the CNS we investigated possible expression of CXCR3 in glial cells. Reverse transcription-polymerase chain reaction experiments and in situ hybridization analysis showed that cultured astrocytes and microglia from both mouse and human sources express CXCR3 mRNA. Protein expression of CXCR3 in both cell types was detected by immunocytochemistry. Moreover, stimulation of cultured glial cells with chemokine ligands for CXCR3 induced intracellular calcium transients and chemotaxis, indicating the functional expression of CXCR3. These results indicate that glial cells in culture functionally express the chemokine receptor CXCR3. Since it has been shown that brain damage rapidly induces expression of neuronal chemokines that activate CXCR3, we suggest that glial CXCR3 might contribute to an intercellular signalling system in the CNS related to pathological conditions. PMID- 12074893 TI - Activation of presynaptic D1 dopamine receptors by dopamine increases the frequency of spontaneous excitatory postsynaptic currents through protein kinase A and protein kinase C in pyramidal cells of rat prelimbic cortex. AB - To determine the effect of dopamine on the frequency of spontaneous excitatory postsynaptic currents (EPSCs) in pyramidal cells of layers V-VI of the prelimbic cortex, whole-cell patch-clamp recordings were made from 92 pyramidal cells of layers V-VI of the rat prelimbic cortex. In normal buffer, dopamine 100 microM apparently increased the frequency of spontaneous EPSCs. Decreasing the concentration of dopamine from 100 to 50 microM was accompanied by a decreased effect of dopamine. Further decreasing the dopamine concentration to 10 and 1 microM had no effects on the frequency of spontaneous EPSCs. In the presence of tetrodotoxin or cadmium, the increasing effect of dopamine was eliminated. The increasing effect of dopamine was blocked by the dopamine D1 receptor antagonist SCH23390, but not by the dopamine D2 receptor antagonist sulpiride. The D1 receptor agonist SKF38393 partially mimicked the increasing effect, but the D2 receptor agonist quinpirole did not. The alpha(1)-adrenoceptor antagonist prazosin could not block the increasing effect of dopamine on the frequency of spontaneous EPSCs in most cells tested. The protein kinase A inhibitor H-89 and the protein kinase C inhibitor chelerythrine could antagonize the effect of dopamine. The protein kinase A activator forskolin and the protein kinase C activator phorbol 12,13-dibutyrate could mimic the effect of dopamine. These results indicate that dopamine, presynaptically acting on dopamine D1 receptors, increases the frequency of spontaneous EPSCs via intracellular protein kinase A and protein kinase C signaling pathways in pyramidal cells of layers V-VI of the prelimbic cortex. PMID- 12074894 TI - Synapse-independent and synapse-dependent apoptosis of cerebellar granule cells in postnatal rabbits occur at two subsequent but partly overlapping developmental stages. AB - It has long been known that cells in the external granular layer die during postnatal development of the cerebellum. More recent findings indicate that at certain developmental stages, cell death occurs upon activation of an apoptotic program. We show that cerebellar granule cells in rabbits undergo programmed cell death at two different stages of maturation. At postnatal day 5 (P5), granule cell precursors and pre-migratory granule cells in the external granular layer incorporate the S-phase markers 5-bromo-2'-deoxyuridine and 5-iodo-2' deoxyuridine with a pattern that is dependent upon the interval between the administration of the two tracers. Within 12-24 h after proliferation a significant number of labeled cells show typical ultrastructural alterations of apoptosis. DNA electrophoresis and cleavage of poly-ADP-ribose polymerase confirm the activation of the apoptotic machinery. After Southern blotting and immunodetection, incorporated 5-bromo-2'-deoxyuridine is present at the level of low size DNA oligomers as soon as 12 h after cell division. Therefore, this apoptotic phase is intrinsic to external granular layer neurons and independent of synaptic interactions with targets.Apoptotic cells, although fewer in number, are detected also in the internal granular layer and tend to increase from P5 to P10. It seems unlikely that these cells undergo DNA fragmentation in the external granular layer and subsequently migrate to their final destination, considering the data on cell cycle kinetics and the rapid tissue clearance by the glia. Parallel fiber-Purkinje spine synapses are already present in the molecular layer at P5. Therefore, the post-migratory granule cells likely undergo apoptosis as a failure to make proper synaptic contacts in the forming molecular layer. We conclude that the massive apoptosis of pre-migratory cells likely has a role in regulating the size of this rapidly expanding population of pre-mitotic neurons. The less tumultuous cell death of post-mitotic granule cells in the internal granular layer appears to be linked to the formation of the mature synaptic circuitry of the developing cerebellar cortex. PMID- 12074895 TI - Innervation of guinea-pig stellate ganglia by nitric oxide synthase, cocaine- and amphetamine-regulated transcript protein- and pituitary adenylate cyclase activating polypeptide-immunoreactive fibers. AB - The present study analyzed using immunohistochemical labeling the distribution and co-localization of nitric oxide synthase (NOS), cocaine- and amphetamine regulated transcript peptide (CARTp) and pituitary adenylate cyclase activating polypeptide (PACAP) with choline acetyltransferase (ChAT)-immunoreactive fibers in the guinea-pig stellate ganglia. ChAT-immunoreactive fibers make pericellular baskets around virtually all stellate ganglia neurons. Pericellular baskets of NOS, CARTp and PACAP fibers were also present around numerous stellate ganglia neurons. Although all the NOS and PACAP fibers also exhibited ChAT immunoreactivity, only some of the CARTp fibers were ChAT-immunoreactive. No evidence of co-localization of NOS, PACAP and CARTp was obtained.These results indicate that NOS, PACAP and CARTp are present in distinct preganglionic axons innervating the guinea-pig stellate ganglia. PMID- 12074896 TI - The importance of seeing it coming: a functional magnetic resonance imaging study of motion-in-depth towards the human observer. AB - Despite their crucial biological relevance, the neural structures differentially activated by the detection of optic flow towards the observer remain to be elucidated. Here, we deploy functional magnetic resonance imaging with normal volunteers to locate the areas differentially activated when motion towards the observer is detected. Motion towards the observer, compared with motion away, showed significant activations (P<0.05, corrected for multiple comparisons), as assessed using statistical parametric mapping, in the lateral inferior occipital cortex bilaterally and in right lateral superior occipital cortex. The areas implicated do not extend into area V5 or subdivisions thereof.Our data suggest that the representations of motion towards the observer implicate perceptual and attentional mechanisms acting at early stages of visual processing in extrastriate cortex. From the standpoint of efficient biological engineering, it makes sense that such crucially important functions as object motion towards the observer would be computed in early visual processing areas. Further studies will be required to determine the extent to which the effects we observed in lateral occipital cortex reflect differential attention to different types of motion, as contrasted with the derivation of explicit representations of motion towards the observer. PMID- 12074897 TI - Attenuation of neuropathic pain by segmental and supraspinal activation of the dorsal column system in awake rats. AB - In addition to its involvement in the transmission of neuropathic pain, the dorsal column system has been shown to have analgesic effects when electrically stimulated. The segmental or supraspinal origin of the analgesia, however, has not been clearly delineated. The aim of this study is to demonstrate the contribution of supraspinal mechanisms to the inhibition of allodynia and hyperalgesia in two different rat models of mononeuropathy. Mononeuropathy was induced, under deep anesthesia, in several groups of rats (n=7 each) following either the chronic constriction injury or the spared nerve injury model. Mechanical and cold allodynia were assessed by the Von Frey monofilaments and by the acetone drop test, respectively. Thermal hyperalgesia was assessed by the paw withdrawal and hot plate tests. Bipolar electrodes for dorsal column stimulation were implanted chronically in all rats on the dorsal aspect of the medulla at the level of the obex. Selective dorsal column bilateral lesions were performed at the upper cervical level in some groups of rats. Dorsal column nuclear stimulation, rostral to selective dorsal spinal lesions, produced strong inhibitory effects on the allodynia and hyperalgesia observed in both models of mononeuropathy. These effects were comparable to those observed following similar stimulations in rats with an intact spinal cord. Our results demonstrate strong inhibitory effects of dorsal column stimulation on neuropathic pain. This inhibition can be attributed to the activation of brainstem pain-modulating centers via rostral projections of the dorsal column nuclei. PMID- 12074898 TI - Interactions between cutaneous afferent inputs to a withdrawal reflex in the decerebrated rabbit and their control by descending and segmental systems. AB - Previous studies have suggested that activation of nociceptive afferents from the heel recruits a supraspinal mechanism, which is modulated by adrenergic descending inhibition, that augments withdrawal reflexes in medial gastrocnemius (MG) motoneurones. To test this idea, we have studied the temporal evolution of reflexes evoked in MG by electrical stimulation of sural nerve A(beta)-, A(delta) and C-fibre axons at 1 Hz, in decerebrated rabbits. Reflexes were analysed in three time bands, estimated to accord to afferent drive from A(beta)- (phase 1), A(delta)- (phase 2) and C-fibre (phase 3) inputs. Stimulation of A(delta)- and C fibres gave significant temporal summation of all reflexes. The alpha(2) adrenoceptor antagonist RX 821002 ((2-(2,3-dihydro-2-methoxy-1,4-benzodioxin-2 yl)-4,5-dihydro-1-H-imidazole)-HCl) (100 microg intrathecal (i.t.)) potentiated, and the alpha(2)-agonist dexmedetomidine (1-30 microg i.t.) depressed all reflexes per se, but the effects of these drugs on temporal summation were secondary to changes in baseline excitability. When C-fibres were stimulated, the N-methyl-D-aspartate (NMDA) receptor antagonist dizocilpine (1 mg i.t.) reduced temporal summation of phase 2 and 3 but not phase 1 reflexes. Spinalisation at L1 in the absence of drugs increased phase 2 and 3 reflexes but had no effect on phase 1, whereas spinalisation after RX 821002 resulted in decreased phase 1 responses with no significant change in later phases. Spinalisation in the presence of dizocilpine resulted in small reductions in phase 3 reflexes only. In all cases spinalisation virtually abolished temporal summation. In spinalised animals, dizocilpine selectively reduced late reflexes, and the opioid antagonist naloxone (100 microg i.t.) augmented all reflexes but gave rise to temporal subtraction of reflexes when C-fibres were stimulated.The present experiments have revealed a number of novel and important features of the sural-MG reflex pathway: (i) activity in fine afferent axons augments the reflexogenic potential of all subsequent afferent input, thereby allowing all afferent drive from the sural field to contribute to withdrawal of the heel; (ii) endogenous adrenergic control of this reflex pathway is completely non-selective; (iii) there is a non adrenergic element of descending inhibition that is selective for the late components of MG reflex responses, and this element is directed particularly against transmission through NMDA receptors; (iv) temporal summation in this reflex is dependent on NMDA receptor-dependent and -independent mechanisms; and (v) this temporal summation is in some way dependent on the integrity of descending pathways. PMID- 12074899 TI - Complexity of sensory environment drives the expression of candidate-plasticity gene, nerve growth factor induced-A. AB - Exposure of animals to an enriched environment triggers widespread modifications in brain circuitry and function. While this paradigm leads to marked plasticity in animals chronically or acutely exposed to the enriched environment, the molecular mechanisms that enable or regulate such modifications require further characterization. To this end, we have investigated the expression profiles of both mRNA and protein products of a candidate-plasticity gene, nerve growth factor induced-A (NGFI-A), in the brains of rats exposed to increased environmental complexity. We found that NGFI-A mRNA is markedly up-regulated throughout the brains of animals exposed to the enriched environment, but not in the brains of either handled-only or undisturbed control groups. The most pronounced effects were observed in the somatosensory and visual cortices, in layers III and V, while more modest increases were observed in all other cortical layers, with the exception of layer I. A striking NGFI-A mRNA up-regulation was also observed in the striatum and hippocampal formation, notably in the CA1 subfield, of animals exposed to the enriched environment paradigm. Immunocytochemistry was also used to investigate the distribution of NGFI-A protein in response to the environmental enrichment protocol. A marked increase in the number of NGFI-A positive nuclei was identified in the enriched environment condition, as compared to undisturbed and handled-only controls, throughout the rat brain. While the greatest number of NGFI-A immunolabeled neurons was found in cortical layers III and V, up-regulation of NGFI-A protein was also detectable in layers II, IV and VI, in both the somatosensory and visual cortices. NGFI-A immunopositive neurons were also more numerous in the CA1 subfield of the hippocampal formation of animals exposed to the enriched environment, but remained at basal levels in both control groups. Our results implicate NGFI-A as one of the possible early genetic signals that ultimately lead to plastic changes in the CNS. PMID- 12074900 TI - Activation of p38 mitogen-activated protein kinase during normal mitosis in the developing retina. AB - The p38 member of the mitogen-activated protein kinase superfamily is engaged by phosphorylation in response to environmental stress signals, and may have either permissive or inhibitor roles upon cell proliferation. The cell cycle in the proliferative zone of the retina is tightly controlled and proceeds in synchrony with interkinetic migration of the neuroblast nuclei. We examined the association of p38 kinase activity with the cell cycle in the normal, non-stressed retina of the developing rat, maintained either in vivo or in vitro. Using immunohistochemistry, we show that mitotic profiles in the developing retina are highly enriched for phosphorylated p38. Blockade of p38 activity with the chemical inhibitor SB203580 for 4 h transiently arrested cells at the metaphase anaphase transition and induced cell death after 20 h. p38 inhibition induced an aberrant mitotic profile, with chromosomes arranged in one side of the cell. The data show that p38 is active during normal mitosis and we suggest that p38 is required for the proper cell cycle progression during metaphase-anaphase transition in retinal neuroblasts. PMID- 12074901 TI - GAP junctional channel inhibition alters actin organization and calcium propagation in rat cultured astrocytes. AB - Astrocytes are connected by gap junctions, which provide intercellular pathways that allow a direct exchange of ions and small metabolites including second messengers and the propagation of electric currents. The roles of gap junctional communication on whole-cell morphology, cytoskeletal organization, and intercellular communication in astrocytes are not yet clear even in vitro, though there are many studies that have examined the active relation between gap junctions and actin filaments in astrocytes. Here we examined the effects of gap junction inhibitors, which do not interrupt the formation but rather the function of gap junctions, on whole-cell morphology, cytoskeletal organization, and intercellular communication in rat cultured astrocytes. Functional blockade of gap junctions during the formation of an astrocytic monolayer resulted in discordance of actin stress fibers between neighboring cells, even though whole cell morphology of these cells did not change by such treatment. Mechanical stimulation-induced calcium wave propagation was significantly reduced in these actin-discordance cells even after thorough wash out. Differentiation of astrocytes in the presence of gap junction inhibitors was associated with morphological disarrangement among neighboring cells due to disordered alignment of actin stress fibers between cells.Our results indicate that gap junctional communication enables cell-to-cell coordination of actin stress fibers in astrocytes, thus enhancing intercellular communication through calcium spread. PMID- 12074902 TI - Role of corticotropin-releasing hormone in the amygdala and bed nucleus of the stria terminalis in the behavioral, pain modulatory, and endocrine consequences of opiate withdrawal. AB - The extra-hypothalamic actions of corticotropin-releasing hormone (CRH) have been accorded an important role in coordinating responses to stressors and contributing to the consequences of drug abuse. Recent proposals suggest that CRH actions in the bed nucleus of the stria terminalis coordinate responses to tonic/unpredictable stressors whereas these actions in the central nucleus of the amygdala coordinate responses to phasic/predictable stressors. We used in situ hybridization histochemistry and site-specific microinjections of a CRH receptor antagonist to study the role of CRH in opiate withdrawal. Rats undergoing opiate withdrawal displayed clear behavioral and autonomic changes accompanied by hyperalgesia and increased plasma corticosterone. In situ hybridization of CRH mRNA revealed significant increases in the central nucleus of the amygdala but not in the bed nucleus of the stria terminalis among rats either chronically pre treated with morphine, given an injection of naloxone, or both (precipitated withdrawal). An increase of CRH mRNA in the paraventricular nucleus of the hypothalamus was specific to rats undergoing withdrawal. Intracerebroventricular microinjection of the CRH receptor antagonist, alpha(h)CRH(9-41), reduced the severity of opiate withdrawal. Microinjections of alpha(h)CRH(9-41) into the central nucleus of the amygdala also reduced the severity of withdrawal whereas bed nucleus of the stria terminalis microinjections of alpha(h)CRH(9-41) were without effect. These experiments provide evidence for a role of amygdala, but not bed nucleus of the stria terminalis, CRH in opiate dependence. We propose a specific role for down-regulation of opiate receptor signaling in increased expression of the CRH gene in the amygdala. Moreover, we suggest that the roles accorded to CRH in the bed nucleus of the stria terminalis versus amygdala in coordinating responses to stressors may need to be reconsidered to distinguish between external and internal/interoceptive stressors. PMID- 12074903 TI - Differential nicotinic receptor expression in monkey basal ganglia: effects of nigrostriatal damage. AB - Our previous work showed that there were marked declines in (125)I-alpha conotoxin MII labeled nicotinic receptors in monkey basal ganglia after nigrostriatal damage, findings that suggest alpha3/alpha6 containing nicotinic receptors sites may be of relevance to Parkinson's disease. We now investigate whether there are differential changes in the distribution pattern of nicotinic receptor subtypes in the basal ganglia in 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP)-lesioned animals compared to controls to better understand the changes occurring with nigrostriatal damage. To approach this we used (125)I-alpha-conotoxin MII, a marker for alpha3/alpha6 nicotinic receptors, and (125)I-epibatidine, a ligand that labels multiple nicotinic subtypes. The results demonstrate that there were medial to lateral gradients in nicotinic receptor distribution in control striatum, as well as ventromedial to dorsolateral gradients in the substantia nigra, which resembled those of the dopamine transporter in these same brain regions. Treatment with MPTP, a neurotoxin that selectively destroys dopaminergic nigrostriatal neurons, led to a relatively uniform decrease in nicotinic receptor sites in the striatum, but a differential effect in the substantia nigra with significantly greater declines in the ventrolateral portion. Competition analysis in the striatum showed that alpha-conotoxin MII sensitive sites were primarily affected after lesioning, whereas multiple nicotinic receptor populations were decreased in the substantia nigra. From these data we suggest that in the striatum alpha3/alpha6 nicotinic receptors are primarily localized on dopaminergic nerve terminals, while multiple nicotinic receptor subtypes are present on dopaminergic cell bodies in the substantia nigra. Thus, if activation of striatal nicotinic receptors is key in the regulation of basal ganglia function, alpha3/alpha6-directed nicotinic receptor ligands may be more relevant for Parkinson's disease therapy. However, nicotinic receptor ligands with a broader specificity may be more important if receptors in the substantia nigra play a dominant role in controlling nigrostriatal activity. PMID- 12074904 TI - Environmental enrichment attenuates cognitive deficits, but does not alter neurotrophin gene expression in the hippocampus following lateral fluid percussion brain injury. AB - Environmental enrichment attenuates neurological deficits associated with experimental brain injury. The molecular events that mediate these environmentally induced improvements in function after injury are largely unknown, but neurotrophins have been hypothesized to be a neural substrate because of their role in cell survival and neural plasticity. Furthermore, exposure to complex environments in normal animals increases neurotrophin gene expression. However, following an ischemic injury, environmental enrichment decreases neurotrophin mRNA levels. Whether these contrasting findings are attributable to differences between injured and uninjured animals or are dependent upon the specific type of brain injury has not been determined. We examined the effects of 14 days of environmental enrichment following a lateral fluid percussion brain injury on behavior and gene expression of brain-derived neurotrophic factor, its high-affinity receptor, TrkB, and neurotrophin-3 in the rat hippocampus. Environmental enrichment attenuated learning deficits in the injured animals, but neither the injury nor housing conditions influenced neurotrophin/receptor mRNA levels. From these data we suggest that following brain trauma, improvements in learning associated with environmental enrichment are not mediated by alterations in brain-derived neurotrophic factor, TrkB or neurotrophin-3 gene expression. PMID- 12074905 TI - Regional decreases in NK-3, but not NK-1 tachykinin receptor binding in dystonic hamster (dt(sz)) brains. AB - Although the pathophysiology of primary dystonias is currently unknown, it is thought to involve changes in the basal ganglia-thalamus-cortex circuit, particularly activity imbalances between direct and indirect striatal pathways. Substance P, a member of the tachykinin family of neuropeptides, is a major component in the direct pathway from striatum to basal ganglia output nuclei. In the present study quantitative autoradiography was used to examine changes in neurokinin-1 (NK-1) and neurokinin-3 (NK-3) receptors in mutant dystonic hamsters (dt(sz)), a well characterized model of paroxysmal dystonia. NK-1 receptors were labeled in 10 dystonic brains and 10 age-matched controls with 3 nM [(3)H] [Sar(9), Met(O(2))(11)]-SP. NK-3 binding sites were labeled in adjacent sections with 2.5 nM [(3)H]senktide. NK-1 binding was found to be unaltered in 27 brain areas examined. In contrast, NK-3 binding was significantly reduced in layers 4 and 5 of the prefrontal (-46%), anterior cingulate (-42%) and parietal (-45%) cortices, ventromedial thalamus (-42%) and substantia nigra pars compacta (-36%) in dystonic brains compared to controls. The latter effects may be particularly relevant in view of evidence that activation of NK-3 receptors on dopaminergic neurons in the substantia nigra pars compacta can increase nigrostriatal dopaminergic activity. Since previous studies indicated that a reduced basal ganglia output in mutant hamsters is based on an overactivity of the direct pathway which also innervates substantia nigra pars compacta neurons, the decreased NK-3 binding could be related to a receptor down-regulation. The present finding of decreased NK-3 receptor density in the substantia nigra pars compacta, thalamic and cortical areas substantiates the hypothesis that disturbances of the basal ganglia-thalamus-cortex circuit play a critical role in the pathogenesis of paroxysmal dystonia. PMID- 12074906 TI - Direct, real-time assessment of dopamine release autoinhibition in the rat caudate-putamen. AB - Inhibition of endogenous dopamine release by photo-released dopamine (i.e., autoinhibition) was characterized in the rat caudate-putamen using combined caged dopamine photolysis and fast-scan cyclic voltammetry. Coronal brain slices (400 microm thick) were perfused with caged-dopamine (150-200 microM in artificial cerebrospinal fluid). Ultraviolet illumination of increasing duration (25-250 ms, approximately 100 microm beam diameter) was focused at the tip of the recording electrode to uncage increasing amounts of exogenous dopamine at the recording sites (0.5-5 microM); a single biphasic electrical stimulus was delivered 0.1-10 s later to induce endogenous dopamine release. The concentrations of both endogenous and exogenous dopamine were determined using voltammetry, thus enabling determination of concentration-dependent inhibition of the endogenous release by the latter. While unaffected by control ultraviolet illumination, endogenous dopamine release was rapidly inhibited by photo-released dopamine in a concentration-dependent manner. Photo-application of 3-5 microM exogenous dopamine inhibited the endogenous release by 90-100% (electrical stimulus applied 1 s after photolysis initiation), an effect prevented by 2 microM sulpiride. The autoinhibition was dependent on the time between photolysis onset and electrical stimulation. Terminal dopamine autoreceptor stimulation led to robust inhibition of endogenous dopamine release with a latency of approximately 200 ms and effective duration of less than 5 s. The percent autoinhibition was a skewed, U shaped function of photolysis/electrical stimulation intervals with the peak inhibition at 1 s. This study directly demonstrates that autoreceptor-mediated inhibition of terminal dopamine release in caudate-putamen is designed to provide a rapid, robust, yet short-lasting modulation of terminal dopamine release. PMID- 12074907 TI - Context-specific spatial representations by lateral septal cells. AB - To test whether the location coding of lateral septal cells is dependent on cue constellations, we examined single units in two different recording arenas on alternating days. Repeated recordings of lateral septal neurons in the same arena revealed that matching locations are encoded on separate days by about one third of the cells. The cells typically showed location-selective firing in only one of the two recording arenas and initially showed unrelated patterns when tested in a different recording arena. When tested for a second time in each recording arena, the initially dissimilar patterns were modified towards increased similarity between arenas. Simultaneously recorded hippocampal principal cells showed distinct place fields for each recording arena throughout the recording sequence. These results indicate that the initial reorganization of the lateral septal location coding may occur as a direct consequence of the hippocampal reorganization. Further septal reorganization is then partially independent of established place fields in the CA1 and CA3 area.Location-selective cells in cortical areas that receive projections from hippocampus proper (i.e. the subiculum and the entorhinal cortex) have not been shown to encode differences between recording arenas. Although some characteristics of this generalized coding scheme have also been found for location-selective lateral septal cells, the encoding of context information was generally preserved in the subcortical target cells of projections from the CA1 and CA3 area. PMID- 12074908 TI - Thalamic neuron apoptosis emerges rapidly after cortical damage in immature mice. AB - In adults and children, head trauma can have long-term neuropathological and functional consequences. The thalamus is a major site of remote neurodegeneration after cortical damage in adult humans and experimental animals, but less is known about thalamic responses to cortical injury in the immature brain. This study introduces an in vivo model of axotomy/target deprivation-induced neuronal apoptosis in the dorsal lateral geniculate nucleus of the thalamus produced by unilateral ablation of the occipital cortex in the immature mouse. We specifically examined whether occipital cortex ablation in the immature brain causes apoptotic death of projection neurons in the dorsal lateral geniculate nucleus. After unilateral occipital cortex aspiration, 10-day-old C57BL/6 mice were recovered for up to 28 days. Fluorogold-prelabeled thalamocortical projection neurons were apoptotic at 36-48 h after ablation. The structural progression of apoptosis in the immature lateral geniculate nucleus reveals typical chromatolytic morphology by 18-24 h, followed by cytoplasmic shrinkage and chromatin condensation characteristic of end-stage apoptosis after 36-48 h. Electron microscopy confirmed the presence of apoptosis. This study shows internucleosomal DNA fragmentation and expression of cleaved caspase-3 occurs rapidly, being noted first at 18 h, well before the peak of apoptotic cell death occurring at 36 h after cortical damage in the immature brain. From these data we suggest that axotomy/target deprivation-induced cell death in the immature brain may: (1) differ from that previously reported in adult mice with respect to the time required for progression to cell death; (2) be mediated by caspase-3 activation. PMID- 12074909 TI - Is novel wheel inhibition of per1 and per2 expression linked to phase shift occurrence? AB - We studied whether access to a novel running wheel in vivo could reset the suprachiasmatic nuclei (SCN) in vitro. Golden hamsters were transferred to dim red light at Zeitgeber time (ZT) 4, given their first exposure to a running wheel for 3 h, and killed at either ZT7 or ZT9. Using a brain slice preparation, the SCN firing rate rhythm in vitro was advanced relative to controls only in the slices prepared at ZT9 (phase shift: 2.36+/-0.06 h, n=4) but not ZT7 (-0.26+/ 0.16 h, n=4). Transitions to dim red light or brain slice preparation at ZT7 or ZT9 alone do not shift the rhythm. Hamsters with wheels had significantly lower levels of SCN per1 mRNA compared with controls at ZT7, and lower per2 mRNA when examined at ZT9. We conclude that 3 h of novel wheel access appears to require some extended time in vivo in order for the SCN to be reset, even beyond the time when per1 mRNA levels have been altered. PMID- 12074910 TI - Examination of the role of the pedunculopontine tegmental nucleus in radial maze tasks with or without a delay. AB - Two radial maze tasks, random foraging and delayed spatial win-shift, have been used to investigate, in rats, the functions and inter-relationships of structures connected through the corticostriatal loops, such as the prelimbic cortex, nucleus accumbens, ventral pallidum and mediodorsal thalamus. The random foraging task is designed to investigate animals' ability to use spatial information to guide foraging on-line. The delayed spatial win-shift task requires, in addition, that animals hold spatially relevant information in working memory across a delay period. The pedunculopontine tegmental nucleus receives direct output from ventral striatal systems and might therefore be expected to share functional properties with them. In the present experiments we have examined the performance of rats bearing bilateral excitotoxic lesions of the pedunculopontine tegmental nucleus on both of these tasks. In acquisition tests rats were given bilateral lesions before any training took place, while in retention tests appropriate training to predetermined criterion levels of performance took place before lesions were made. In both tasks, and in both acquisition (no prelesion training) and retention (prelesion training) tests, rats with pedunculopontine lesions made significantly more errors in selecting arms to enter than did control rats. There was no motor impairment present in pedunculopontine tegmental nucleus-lesioned rats - on the contrary, on measures of speed (latency to make first arm choice and the mean time for subsequent choices) pedunculopontine-lesioned rats were slightly faster than control rats. We suggest that the pedunculopontine tegmental nucleus shares functional properties with frontostriatal systems and that it forms part of a brainstem-directed stream of striatal outflow different to the cortical re-entrant system via the thalamus. PMID- 12074911 TI - Isolation rearing in the rat disrupts the hippocampal response to stress. AB - Both human schizophrenia and the effects of isolation rearing in rats produce deficits in hippocampal and cortical functioning. This study was concerned with identifying changes associated with altered neuronal function in the rat hippocampus following isolation rearing. Rats were isolated from weaning at 21 days postnatal for 6 weeks and the hippocampal sensitivity to isolation rearing and stress were studied using c-fos immunohistochemistry and in vivo microdialysis. Isolation rearing altered neuronal activity measured by Fos-like immunoreactivity in the specific brain areas as measured by either increased or reduced expression. Basal neuronal activity in the ventral CA1 hippocampus in isolation-reared rats was notably higher compared to group-reared rats but markedly lower Fos-like immunoreactivity was found in the central and basolateral nuclei of the amygdala. Exposure to stress produced differential effects on neuronal activity in isolation-reared rats between the dorsal and ventral hippocampus, with increased Fos-like immunoreactivity in the dorsal hippocampus but lower Fos-like immunoreactivity in the ventral hippocampus compared to group reared rats. These results indicate that isolation rearing may alter the relationship between hippocampal neuronal function in the dorsal and ventral hippocampus. An in vivo microdialysis study showed that systemically administered parachloroamphetamine (2.5 mg/kg, i.p.) enhanced extracellular 5 hydroxytryptamine (5-HT) in the dorsal hippocampus in group-reared but not in isolation-reared rats. Restraint stress had no effect on hippocampal extracellular 5-HT in group-reared rats but reduced levels in isolation-reared rats during the period of restraint. Inescapable mild footshock produced a marked increase in extracellular hippocampal 5-HT in group-reared but not isolation reared rats. Overall the results provide extensive evidence that isolation rearing alters presynaptic 5-HT hippocampal function and that the neuronal response to stress is altered by isolation. Isolation rearing in the rat alters hippocampal function, including the serotonergic system, leading to changes in neurotransmitter systems in other brain areas. These changes may model aspects of human neurodevelopmental disorders such as schizophrenia. PMID- 12074912 TI - The mechanisms of neuronal death produced by mitochondrial toxin 3-nitropropionic acid: the roles of N-methyl-D-aspartate glutamate receptors and mitochondrial calcium overload. AB - Previous studies showed that 3-nitropropionic acid, an irreversible inhibitor of succinate dehydrogenase, produced neuronal death secondary to perturbed intracellular calcium homeostasis. However, the response of intramitochondrial calcium ([Ca(2+)](m)) to 3-nitropropionic acid remains unknown. In this study, we investigated the roles of and relationships among [Ca(2+)](m) overload, mitochondrial reactive oxygen species, and mitochondrial membrane depolarization in 3-nitropropionic acid-induced neuronal death. Following 1 mM 3-nitropropionic acid treatment on primary rat neuronal cultures, there was a gradual increase of [Ca(2+)](m) beginning at 2-4 h post 3-nitropropionic acid application, and a twofold increase of mitochondrial reactive oxygen species at 4 h. These were followed by mitochondrial membrane depolarization at 6-8 h post-treatment. By inhibiting [Ca(2+)](m) uptake, Ruthenium Red attenuated the production of reactive oxygen species, and prevented the 3-nitropropionic acid-induced mitochondrial membrane depolarization and 70% of apoptotic neuronal death (P<0.001). Inhibition of caspase activation attenuated the elevation of [Ca(2+)](m) (P<0.001), indicating that caspase activation plays a role in the elevation of [Ca(2+)](m). MK-801, an antagonist of N-methyl-D-aspartate (NMDA) glutamate receptors, prevented 3-nitropropionic acid-induced [Ca(2+)](m) elevation, caspase-3 activation, mitochondrial depolarization, and neuronal death. We conclude that the activation of NMDA glutamate receptor contributes to mitochondrial alterations induced by 3-nitropropionic acid. Inhibition of its activation and [Ca(2+)](m) overload with subsequent mitochondrial membrane depolarization can therefore attenuate the neuronal death induced by 3 nitropropionic acid. PMID- 12074913 TI - Spatiotemporal analysis of signal transducer and activator of transcription 3 activation in rat brain astrocytes and pituitary following peripheral immune challenge. AB - The host response to peripheral inflammation induces fever and behavioural depression that are supposed to be centrally mediated by cytokines. Several proinflammatory cytokines activate 'signal transducer and activator of transcription' 3 (STAT3) via gp130-like receptor signaling. In order to determine which cells in the rat brain and pituitary are activated during bacterial inflammation, we investigated in a spatiotemporal manner the activation of STAT3 in these organs following peripheral lipopolysaccharide (LPS) challenge. Under basal conditions, STAT3 immunoreactivity was observed in neurones and some glial cells throughout the brain. Two hours after the administration of LPS, nuclear localisation of STAT3 (hallmark of activation) was observed in zones at the interface between brain and blood or cerebrospinal fluid such as pituitary, ependymal layer, meninges, glia limitans, circumventricular organs and surrounding nervous parenchyma. Four hours after LPS, the nuclear activation of STAT3 propagated to cells located inside the parenchyma (cortex, hypothalamus, corpus callosum and hippocampus among others) and declined 8 h after treatment. Double labelling of STAT3 and glial fibrillary acidic protein identified activated cells in the parenchyma as astrocytes. These data show that STAT3 is activated in the pituitary and in brain astrocytes after a peripheral LPS challenge as demonstrated by immunohistochemistry. Astrocytes may therefore play a key role in the brain response to peripheral inflammation. PMID- 12074914 TI - Intracerebroventricular but not intravenous interleukin-1beta induces widespread vascular-mediated leukocyte infiltration and immune signal mRNA expression followed by brain-wide glial activation. AB - Interleukin-1beta (IL-1beta) is a pro-inflammatory cytokine that appears in brain and cerebrospinal fluid following peripheral immune challenges and central infections or injury. We examined the consequences of i.c.v. infusion of IL-1beta on mRNA expression of several immune markers and on recruitment of peripheral leukocytes. Awake rats were infused with IL-1beta (100 ng/rat) into the lateral ventricle, and 0.5, 2, 4, 8, 12, or 24 h later, animals were killed and their fresh-frozen brains processed for in situ hybridization and immunohistochemistry. Widespread vascular expression of inhibitory factor kappa(B)alpha (Ikappa(B)alpha, marker of nuclear factor kappa(B)alpha transcriptional activity) and inducible cyclooxygenase (COX-2) mRNAs at 0.5-2 h was credited to movement of IL-1beta along ventricular, subarachnoid, and perivascular pathways to target endothelia that express type 1 IL-1 receptor mRNA. Induction of monocyte chemoattractant protein-1 mRNA and intercellular adhesion molecule-1 (ICAM-1) immunostaining on endothelia began at 0.5-2 h. Leukocytes (neutrophils and monocytes, recognized by morphology and CD45 and ED1 immunostaining) appeared in meninges and blood vessels at 2-4 h and diffusely penetrated the parenchyma at 8 24 h. The leukocytes strongly expressed IL-1beta and inducible nitric oxide synthase mRNAs. Beginning at 4-12 h, astrocytes (glial acidic fibrillary protein mRNA and protein and c-fos mRNA) and microglia (ionized calcium-binding adaptor molecule 1 mRNA and protein) showed widespread activation. Other rats received i.v. IL-1beta (6 microg/kg). Their brains showed induction of Ikappa(B)alpha and COX-2 mRNAs in the vasculature at 2 h but none of the other sequelae. In summary, our data indicate that IL-1beta in the cerebrospinal fluid reaches its target receptors on the endothelia via perivascular volume transmission, up-regulates ICAM-1, and triggers a targeted leukocyte emigration and widespread glial activation stimulated perhaps by pro-inflammatory molecules expressed by leukocytes. The dramatic difference between i.c.v. and i.v. routes of administration underscores the potency of IL-1beta within the brain to dynamically affect the cellular trafficking component of 'immune privilege'. PMID- 12074915 TI - High-resolution spiral computed tomography with multiplanar reformatting, 3D surface- and volume rendering: a non-destructive method to visualize ancient Egyptian mummification techniques. AB - Ancient Egyptians used mummification techniques to prevent their deceased from decay. This study evaluates the potential of computed tomography (CT) in determining these techniques in a non-destructive way. Twenty-five mummies were studied by using high-resolution spiral CT, 1mm slice thickness for the head and 3mm slice thickness for the rest of the body. Images were reconstructed with 3D, multiplanar reformatting and volume rendering. In all cases the used mummification techniques could be reconstructed. The way the brain was removed, the presence of chemicals, like resin and natron, could be detected and the way the intestines were handled could be made visible. The use of CT is indispensable as a non-destructive method in the reconstruction of mummification techniques. PMID- 12074916 TI - Technical innovation of cardiac multirow detector CT using multisector reconstruction. AB - Multisector reconstruction is a newly developed algorithm for multirow detector CT in cardiac study. Using volume data sets obtained by ECG-gated scanning, we can reconstruct cardiac images at any desired phase of the cardiac cycle retrospectively. In retro-processing multiplanar and three-dimensional images, thin-slice images with overlapping increment have a great advantage due to increasing z-axis resolution. In this article, we present principles of the algorithm, a phantom study, clinical applications and perspectives for the future. PMID- 12074917 TI - Geometric reconstruction for computational mesh generation of arterial bifurcations from CT angiography. AB - A methodology for patient-specific model reconstruction and computational mesh generation of arterial bifurcations from angio-CT scans is presented. Three dimensional models were reconstructed with a level set technique, analyzed with a skeletoning algorithm and automatically decomposed into branches. Cooper scheme was then employed to generate high quality hexahedral mesh. We successfully applied our technique to the carotid bifurcations of two patients affected by severe atherosclerotic plaques. The proposed technique is fast, accurate and reproducible, and can be a useful tool for the evaluation of arterial fluid dynamics within conventional computed tomography investigations. PMID- 12074918 TI - A computer model of lung morphology to analyze SPECT images. AB - Measurement of the spatial distribution of aerosol deposition in human lungs can be performed using single photon emission computed tomography (SPECT). To relate deposition patterns to real lung structures, a computer model of the airway network has been developed. Computer simulations are presented that are compatible with the analysis of SPECT images. Computational techniques that are consistent with clinical procedures are used to analyze airways by type and number within transverse slices of the lung volume. The computer models serve as customized templates, which when analyzed alongside gamma scintigraphy images, can assist in the interpretation of human test data. PMID- 12074919 TI - Localized renal cystic disease: imaging findings, pathologic correlation, and management approach. AB - A case of unilateral renal cystic disease of the kidney is reported, in which the cysts were located to the lower pole of the right kidney. The radiologic and pathologic findings are described and their differential diagnosis is discussed. Conservative treatment and imaging follow-up is proposed for this condition. PMID- 12074920 TI - Structural MR image processing using the BRAINS2 toolbox. AB - Medical imaging has opened a new door into biomedical research. In order to study various diseases of the brain and detect their impact on brain structure, robust and user friendly image processing packages are required. These packages must be multi-faceted to distinguish variations in size, shape, volume, and the ability to detect longitudinal changes over the course of an illness. This paper describes the BRAINS2 image processing package, which contains both manual and automated tools for structural identification, methods for tissue classification and cortical surface generation. These features are described in detail, as well as the reliability of these procedures. PMID- 12074921 TI - Diffusion MRI findings in isolated intracranial angiitis. AB - This paper reports an 8-year-old girl with proven primary (isolated) angiitis of the central nervous system. On diffusion MRI, multiple scattered lesions were noted in the internal capsulas, thalami, and in the left middle cerebellar pedincle. These were hyperintense on b=1000s/mm(2) (true diffusion) images. The apparent diffusion coefficient (ADC) values of these ranged between 0.40 and 0.52 x 10(-3)mm(2)/s, consistent with acute ischemic infarction (cytotoxic edema). The ADC values of a relatively old lesion in the left occipital region ranged between 1.65 and 1.82 x 10(-3)mm(2)/s, consistent with chronic infarction. PMID- 12074922 TI - Proton MR spectroscopy in Rett syndrome. AB - Seven patients (mean age 7.7yr) with Rett syndrome, a condition with progressive regression of psychomotor development are included in this study. Proton MR spectroscopy images were obtained with the multivoxel chemical-shift imaging mode (TR=1500ms, TE=40ms). Spectra from 224 voxels in the brain parenchyma were studied. N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), and myoinositol (mI) peaks were quantitatively evaluated, and NAA/Cr, NAA/Cho, and Cho/Cr, mI/Cr ratios were calculated. Five age-matched normal cases were available as controls. In three patients with Rett syndrome spectroscopy findings were normal, and the metabolite ratios were similar to control cases. In the remaining four patients with the syndrome prominent decrease of the NAA peak was the main finding resulting in decreases in NAA/Cr (1.14+/-17), and NAA/Cho (1.08+/-27) ratios (p<0.0001). Cho/Cr ratios (0.93+/-26), and mI/Cr ratios (0.88+/-36) were normal compared to controls. There was no correlation between spectroscopic changes and clinical status of the patients. The findings suggested that not only reduced neuronal-dendritic arborizations but also decreased neuronal function could contribute to spectroscopy changes in Rett syndrome. PMID- 12074923 TI - Global optimization of mutual information: application to three-dimensional retrospective registration of magnetic resonance images. AB - A global optimization technique for image registration, based on mutual information, that can be used in conjunction with a multi-resolution paradigm is described. This technique combines genetic algorithm in continuous space, which is a stochastic method and is very efficient in large search space, with dividing rectangle, which is a deterministic method that theoretically guarantees global optimization and is efficient in small search space. Calculations were performed for determining the optimum parameters for implementing this method. This technique was applied to register magnetic resonance images of brain. For comparison, the registration results using AIR, a commonly employed software package, are presented. PMID- 12074924 TI - Ontogeny of immunoglobulin expression in the brushtail possum (Trichosurus vulpecula). AB - Marsupials, unlike eutherians, are born immunologically immature, without circulating lymphocytes or organised lymphoid tissue. Their immune response develops while they are in the pouch not in the uterus. In this study, the onset time of immunoglobulin expression in Trichosurus vulpecula pouch young was estimated by reverse transcription polymerase chain reaction. As in eutherian species, IgM heavy chain transcripts were detected first, at day 10 post partum. The first switched transcript, detected at day 18, was Calpha. Cgamma and Cvarepsilon transcripts were not present at day 72, but were seen at day 103, approximately corresponding to the time of release of the teat and exposure to new antigens, as well as the time of the loss of capacity to absorb maternal Igs through the gut. PMID- 12074925 TI - Thioester function is conserved in SpC3, the sea urchin homologue of the complement component C3. AB - The amino acid sequence of the thioester site in the alpha chain of SpC3, the sea urchin homologue of C3, is conserved. This implies a conserved function of covalent bond formation with amine or hydroxyl groups on target molecules. When coelomic fluid (CF) was incubated with 14C-methylamine, a classic assay for thioester binding function, the alpha chain became labeled. When CF was treated to induce autolysis, peptide bond cleavage occurred at the thioester site. Autolysis could be blocked or reduced by pre-treating CF with either methylamine or yeast, both of which are known to bind to thioester sites C3 proteins from other organisms. The data suggest that SpC3 can bind to target cell surfaces, constituting indirect evidence that it can covalently bind to pathogen surfaces and function as an opsonin in vivo. This activity may be an important aspect of host defense in the sea urchin. PMID- 12074926 TI - Morphological consequences for multi-partner chimerism in Botrylloides, a colonial urochordate. AB - Studies on multi-partner chimeras (MC) of the colonial urochordate Botryllus schlosseri have revealed that these chimeras form more stable and vigorous entities than bi-partner chimeras (BC). This outcome has been further studied here on another botryllid species. Botrylloides leachi subpopulation 1, by analysing the morphological consequences of BC, chimeras comprising three partners, tri-chimeras (TC) and chimeras comprising six partners each, hexa chimeras (HC). For each chimerical type, five different genotypic combinations with three replicates per combination were established. Chimeras were observed for up to 10 months, at which point, all had died. While life spans of BC, TC and HC were the same, the average maximum sizes of HC were higher resulting from more than three times greater daily growth rate. BC and TC reached a maximal chimeric size at a much earlier age. Some morphological resorptions were also expressed differently in HC as compared to BC. When comparing chimerical parameters of bi- vs. multi-chimerism between Botryllus and Botrylloides, these two genera differ in nine out of 13 characteristics tested. However, it seems that irrespective to the species studied and the different characteristics expressed, MC in botryllid ascidians is a commonly developed phenomenon shaped by evolutionary pressures that interact on a 'group' level instead of each individual partner. MC reveal modified entities in which contradicting intraspecific interactions are alleviated. PMID- 12074927 TI - Dexamethasone-induced apoptosis in immune cells from peripheral circulation and lymphomyeloid tissues of juvenile clearnose skates, Raja eglanteria. AB - Juvenile clearnose skates (Raja eglanteria) were injected intramuscularly with dexamethasone-21-phosphate at 50, 75, and 100mg/kg body weight. After 24h, skates were sacrificed and lymphomyeloid tissues (thymus, spleen, Leydig organ, and epigonal organ) were removed and whole blood was sampled. Tissues were used fresh for imprints or prepared for histology by solvent fixation or freezing in liquid nitrogen. Apoptosis in fixed tissues was assessed by transmission electron microscopy. Frozen sections and cytospin preparations of peripheral blood leukocytes (PBL) were evaluated by the TUNEL reaction to detect DNA strand breaks. Dexamethasone treatment increased apoptotic activity in all lymphomyeloid tissues as well as in PBL. These studies demonstrate that immune cells of elasmobranchs have the capacity for glucocorticoid-driven apoptosis, and that programmed cell death as a mechanism to regulate immune cell production appears to have been conserved during vertebrate evolution. PMID- 12074928 TI - Characterisation of immunoglobulin light chain cDNAs of the Atlantic salmon, Salmo salar L.; evidence for three IgL isotypes. AB - By screening a cDNA library and analysis of DNA produced by a combined 3'RACE/5' anchored PCR, we have isolated three isotypes of IgL in the Atlantic salmon. Two of the isotypes were homologous to rainbow trout IgL1 and L2 sequences, while the third represents a previously uncharacterised salmonid IgL. The novel type 3 CL region is homologous to spotted wolffish c1 and yellowtail sequences, while the VL region is more similar to channel catfish F class than to any other fish VL sequences. Southern analysis indicates that the gene segments of all three isotypes are organised in multiple clusters. In addition, the VL gene segments of type 3 are arranged in opposite orientation relative to the JL and CL segments, while gene segments in type 2 clusters are all in the same orientation. Although transcripts of type 1 and 3 were readily found in the spleen and head kidney, only minute amounts of type 2 transcripts were seen. The majority of type 3 messages were truncated, suggesting that spliced and full-length transcripts of this isotype probably are present at a low level compared to type 1 transcripts. The uniqueness of the type 3 VLJL sequences suggests that this isotype offers additional diversity to the antigen-binding site of Atlantic salmon immunoglobulins. PMID- 12074929 TI - Immunization of goldfish with excretory/secretory molecules of Trypanosoma danilewskyi confers protection against infection. AB - Trypanosoma danilewskyi is a protozoan that lives in blood and other tissues of fish. In the aquaculture industry, economic losses may be substantial, since the prevalence of infection may approach 100% and the parasite may cause significant mortality in farmed carp. Most of the surviving fish acquire resistance after elimination of the primary infection. In this study, we examined whether protection against infection could be induced in naive goldfish immunized with excretory-secretory (ES) products of the parasite. The ES extracts were administered in conjunction with Freund's complete or incomplete adjuvant (FCA and FIA, respectively). Parameters used to assess the efficacy of immunization after challenge infection, included prevalence of infection, abundance of parasites, and presence of parasite-specific antibodies. Intraperitoneal inoculation of ES products in FCA conferred protection against T. danilewskyi infection (P<0.05). Administration of ES products (with or without FIA) conferred insignificant levels of protection. In an attempt to identify the immunogenic ES molecules, we assessed whether anti-parasite antibodies present in the serum collected from experimentally infected fish or rabbits immunized with ES recognized parasite ES antigens. An immunoblot using rabbit anti-parasite antibody revealed a recognition profile of molecules (two antigens of approximately 102-104 and 70-72kDa) similar to that of immune goldfish serum. While the antigens that confer protection need further molecular characterization, our results suggest that the administration of ES products may allow for the design of control strategies for T. danilewskyi. PMID- 12074930 TI - An alternative internal splicing site defines new Ikaros isoforms in Pleurodeles waltl. AB - The Ikaros gene encodes a family of transcription factors which plays a crucial role in hematopoiesis. To improve our knowledge about the immune system of Pleurodeles waltl, we sequenced the cDNA coding for the Ik-1 isoform of that salamander and analyzed its tissue expression by semi-quantitative RT-PCR. Ikaros transcripts are abundant in the thymus and the spleen, thereby confirming that these organs are, respectively, the primary and secondary lymphoid tissues of Pleurodeles. Analysis of the isoforms produced by this animal revealed two isoforms characteristic of amphibians in which an alternative internal splicing site deletes the 3' half of exon 3 which interestingly comprises the first Zn finger of Ikaros. Ikaros transcripts were found at the earliest stages of development of Pleurodeles indicating that Ikaros has a function at the very early lymphopoietic stages. Moreover, the presence of Ikaros transcripts in spermatozoa suggests that this protein could have another and yet unknown function. PMID- 12074931 TI - Priming of dolphin neutrophil respiratory burst by recombinant tumor necrosis factor alpha. AB - We studied the effects of recombinant dolphin tumor necrosis factor alpha (rdoTNFalpha) on the respiratory burst activity of dolphin neutrophils. rdoTNFalpha enhanced the luminol-dependent chemiluminescence response of dolphin neutrophils induced by concanavalin-A, opsonized zymosan, and heated plasma, but not that induced by phorbol myristate acetate. The TNF-associated priming activity was concentration- and preincubation time-dependent, and heat-instable. These data suggest that, as in human neutrophils, TNFalpha enhances the respiratory burst in dolphin neutrophils that follows short-term incubation with various receptor-mediated agonists. PMID- 12074932 TI - Chlorodinitrobenzene-mediated damage in the human erythrocyte membrane leads to haemolysis. AB - 1-chloro-2,4-dinitrobenzene (CDNB), an intracellular glutathione-depleting agent, has been shown to have an adverse effect on erythrocyte membrane integrity. In the current study, we have demonstrated that CDNB caused haemolysis of human red blood cells (RBC) at higher concentrations (>or= 5 mM). The haemolysis induced by CDNB was preceded by the leakage of K(+) from the cells suggesting the colloid osmotic nature of this lysis. The inclusion of molecules of increasing size in the extracellular media inhibited both the rate and extent of haemolysis thus supporting the proposal of CDNB-induced pore formation. The size of membrane lesions increased with an increase in the concentration of CDNB. SDS-PAGE demonstrated that CDNB causes the polymerisation and/or fragmentation of membrane proteins. Although CDNB has been shown to cause a drastic reduction in membrane thiols, our data suggest that the CDNB-induced formation of membrane disulfide bonds as a prima facie cause of permeability enhancement is unlikely. PMID- 12074933 TI - Direct antimicrobial properties of substance P. AB - The structural similarity between substance P (SP, Arg-Pro-Lys-Pro-Gln-Gln-Phe Phe-Gly-Leu-Met-NH(2)) and Arg/Pro rich bactericidal peptides suggests a possible direct effect of SP on invasive microbes. We now present evidence that substance P possesses direct antimicrobial activity, highest against S. aureus. A substance P antagonist also possesses such activity but while less potent than substance P agonist S. aureus, is more potent than substance P against C. albicans. Our data also show that the endogenous peptides bradykinin and neurotensin, that also play role in modulation of the host-defense system in situ, have antimicrobial properties but are less potent than substance P. PMID- 12074934 TI - Age-related decline of serotonin transporters in living human brain of healthy males. AB - There is growing interest in serotonin transporter (5-HTT) function in the human brain, since alteration in 5-HTT has been suggested in a variety of neurophychiatric disorders. Age-related decline in postsynaptic 5-HT receptors has been demonstrated in postmortem human studies and in vivo imaging studies, and has been assumed to be related to changes in mental function in the normal aging process. However, few studies have investigated the aging effect on 5-HTT in human brain in vivo, since the availability of suitable ligands has been limited. To investigate the aging effect on 5-HTT in living human brain, we performed positron emission tomography (PET) scans with a selective ligand for 5 HTT, [11C](+)McN5652. We examined 28 healthy male volunteers aged between 20 and 79 years. The uptake was quantified in the thalamus and midbrain by graphical analysis with the cerebellum as a reference tissue, and binding potential (BP) was used for the index of 5-HTT binding. There was a significant age-related decline in BP in the thalamus and midbrain. The decline in [11C](+)McN5652 binding was 9.6% per decade in the thalamus and 10.5% per decade in the midbrain. PMID- 12074935 TI - In vivo effects of Panax ginseng extracts on the cytochrome P450-dependent monooxygenase system in the liver of 2,3,7,8-tetrachlorodibenzo-p-dioxin-exposed guinea pig. AB - The effects of the subchronic administration of Panax ginseng extracts were examined on the hepatic cytochrome P450-dependent monooxygenase system of guinea pigs pre-exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Panax ginseng extracts were intraperitoneally administered to guinea pigs at 100 mg/kg/day for 14 days from 1 week after a single intraperitoneal injection of 1 microg of TCDD/kg of body weight. TCDD treatment increased the total cytochrome P450 content 2.86-fold, and this was remarkably inhibited by the administration of Panax ginseng extracts. Treatment with ginseng extract alone also decreased the contents of cytochrome P450 by 33%, but both TCDD and ginseng extracts had no effect on cytochrome b(5) content. The administration of TCDD resulted in a 1.73 fold increase in microsomal NADPH-cytochrome P450 reductase activity in the guinea pig liver, and this was significantly inhibited by ginseng extracts, but treatment with ginseng extracts alone had no effect on its activity, and no statistical changes in the activity of NADPH-cytochrome b(5) reductase were observed in guinea pig liver due to TCDD and/or ginseng extract administration. Compared to the control, ECOD activity remarkably (1.76-fold) increased after TCDD administration, but this increase was completely inhibited by treatment with ginseng extract. Treatment with ginseng extract alone resulted in a 50% reduction of ECOD activity. TCDD administration remarkably induced benzphetamine demethylation (BPDM) activity, while ginseng extract also slightly increased the enzyme's activity, but the induction attributed to ginseng extracts was not statistically significant. Even though administration of ginseng extracts slightly inhibited TCDD-induced BPDM activity, the inhibition was not statistically significant. These results indicate that ginseng extract exerts different effect on the induction of P450 isozymes. From these results, we suggest that Panax ginseng extracts may act as an inhibitor of CYP1A rather than that of CYP2B. PMID- 12074936 TI - Pretreatment of CD-1 mice with 4-methylpyrazole blocks toxicity from the gamma hydroxybutyrate precursor, 1,4-butanediol. AB - 1,4-Butanediol (1,4-BD) is the dihydroxy precursor of gamma-hydroxybutyrate (GHB), a popular recreational drug that has been banned by the United States Food and Drug Administration (FDA) and controlled as a federal schedule I drug. 1,4-BD is enzymatically converted in vivo to GHB by alcohol dehydrogenase (ADH), and overdoses can result in coma, severe respiratory depression, bradycardia, hypothermia, seizures, and death. Presently, there is no antidote. We pretreated CD-1 mice with the ADH antagonist, 4-methylpyrazole (4-MP), to determine if blocking ADH can prevent or decrease toxicity from 1,4-BD overdose. Pretreatment with 4-MP increased the Toxic Dose-50 (TD(50)) of 1,4-BD for the righting reflex from 585 mg/kg (95% CI, 484-707 mg/kg) in control mice to 5,550 mg/kg (95% CI, 5,353-5,756 mg/kg) in pretreated mice. Pretreatment with 4-MP also increased the TD(50) of 1,4-BD for the rotarod test from 163 mg/kg (95% CI, 136-196 mg/kg) in control mice to 4,900 mg/kg (95% CI, 4,812-4,989 mg/kg) in pretreated mice. Pretreatment with 4-MP significantly decreased the toxicity of 1,4-BD in CD-1 mice, presumably by inhibiting its ADH biotransformation to GHB. 4-MP warrants further investigation as a potential antidote for this increasingly abused drug. PMID- 12074937 TI - Complementary effects of adenosine and angiotensin II in hypoxemia-induced renal dysfunction in the rabbit. AB - The acute renal effects of hypoxemia and the ability of the co-administration of an angiotensin converting enzyme inhibitor (perindoprilat) and an adenosine receptor antagonist (theophylline) to prevent these effects were assessed in anesthetized and mechanically-ventilated rabbits. Renal blood flow (RBF) and glomerular filtration rate (GFR) were determined by the clearances of para aminohippuric acid and inulin, respectively. Each animal acted as its own control. In 8 untreated rabbits, hypoxemia induced a significant drop in mean blood pressure (-12 +/- 2%), GFR (-16 +/- 3%) and RBF (-12 +/- 3%) with a concomitant increase in renal vascular resistance (RVR) (+ 18 +/- 5%), without changes in filtration fraction (FF) (-4 +/- 2%). These results suggest the occurrence of both pre- and postglomerular vasoconstriction during the hypoxemic stress. In 7 rabbits pretreated with intravenous perindoprilat (20 microg/kg), the hypoxemia-induced changes in RBF and RVR were prevented. FF decreased significantly (-18 +/- 2%), while the drop in GFR was partially blunted. These results could be explained by the inhibition of the angiotensin-mediated efferent vasoconstriction by perindoprilat. In 7 additional rabbits, co-administration of perindoprilat and theophylline (1 mg/kg) completely prevented the hypoxemia induced changes in RBF (+ 11 +/- 3%) and GFR (+ 2 +/- 3%), while RVR decreased significantly (-14 +/- 3%). Since adenosine and angiotensin II were both shown to participate, at least in part, in the renal changes induced by hypoxemia, the beneficial effects of perindoprilat and theophylline in this model could be mediated by complementary actions of angiotensin II and adenosine on the renal vasculature. PMID- 12074938 TI - Chemical sympathectomy alters food intake and thermogenic responses to catecholamines in rats. AB - It has been suggested that the sympathetic nervous system contributes to the short-term control of feeding. The adrenergic innervation of some splanchnic organs seems to be especially involved in such processes, since catecholamines reduce feeding only when injected intraperitoneally or intraportally. In this work, the effects of neonatal sympathetic denervation with guanethidine (Gnt) upon food intake were assessed in adult rats. Gnt-treated male rats had lower body weight gain. The hypophagic response to intraperitoneal (ip) norepinephrine was 70% higher in Gnt-treated animals as compared to controls (P < 0.05); that of epinephrine (E) by 33% (P < 0.05) and that of isoproterenol was not significantly modified. As in normal rats, the hypophagic effect was much stronger after ip than after intramuscular (im) administration (P < 0.05). On the other hand, resting oxygen consumption (VO2) was consistently lower in denervated animals. Ip E administration did not modify VO2, while im E caused increased motor activity and VO2 (P < 0.05). In contrast to control rats, the respiratory exchange ratio in ad libitum fed Gnt rats did not decrease after Ip E administration, suggesting a lack of effect upon lipid mobilization. The lower rate of body weight gain induced by neonatal Gnt sympathectomy might be due to lower daily food intake possibly related, in part, to the sensitization of the alpha-adrenergic porto hepatic response to endogenous catecholamines. Compared with controls, Gnt treated rats also showed a limited thermogenic capacity not related to feeding, and a greater degree of carbohydrate oxidation, possibly due to a defect in E induced lipolysis, which is beta-adrenergic. PMID- 12074939 TI - Effects of magnesium sulfate on energy metabolites and glutamate in the cortex during focal cerebral ischemia and reperfusion in the gerbil monitored by a dual probe microdialysis technique. AB - Previous studies have demonstrated that magnesium sulfate has cytoprotective properties for treating experimental rat brain injuries. The aim of this study is to evaluate changes in energy-related metabolites and glutamate in the cortex of gerbils subjected to focal cerebral ischemia with the pretreatment of magnesium sulfate. The focal cerebral ischemia was produced by the occlusion of the right common carotid artery and the right middle cerebral artery for 60 mins. A significant decrease in infarct size was found in the magnesium sulfate treated group when compared to the controls. Two microdialysis probes were inserted bilaterally into the cortex to monitor extracellular glucose, lactate, pyruvate and glutamate during cerebral ischemia and reperfusion periods. The present study showed a dynamic decrease of glucose (10% of the baseline), pyruvate (15% of the baseline), and an increase of lactate (200% of the baseline) and glutamate (1400% of the baseline) on the ipsilateral side during ischemia in the control group. Magnesium sulfate significantly preserved glucose (up to 50% of the baseline) and pyruvate (70% of the baseline) levels in the ipsilateral side during ischemia. There was significant attenuation in the elevation of glutamate and lactate (500% and 150% of the baseline, respectively) when treatments of magnesium sulfate were administered. No significant influence on these neurochemicals in the contralateral side was observed in either group. These results suggest that both the preservation of cellular energy metabolism, and the attenuation of glutamate release during cerebral ischemia and after restoration of reperfusion may contribute to the neuroprotective effects of magnesium sulfate. PMID- 12074940 TI - Endogenous expression of nNOS protein in several neuronal cell lines. AB - Several neuronal cell lines were screened for endogenous expression of neuronal nitric oxide synthase (nNOS) protein using Western blot analysis. Detectable levels of the nNOS protein were evident in the SK-N-SH, SH-SY5Y, and N1E-115 neuroblastoma cell lines, as well as the NG108-15 neuroblastoma x glioma hybrid. Only trace amounts were visible in Neuro2A human neuroblastoma cells. The presence of endogenously expressed nNOS in these cells may allow for the study of the interaction between nNOS and the endogenous receptor systems expressed in the same cells. PMID- 12074941 TI - Involvement of the nitric oxide/L-arginine and sympathetic nervous systems on the vasodepressor action of human urotensin II in anesthetized rats. AB - This study examined if the nitric oxide (NO)/L-arginine pathway participates in and if the sympathetic nervous system attenuates the depressor action of human urotensin II. I.V. bolus injections of human urotensin II (0.1-30 nmol/kg) caused dose-dependent decreases in mean arterial pressure (MAP, EC(50) = 2.09 +/- 0.8 nmol/kg; Emax = -18 +/- 3 mmHg ) and increases in heart rate. The depressor response to human urotensin II (3 nmol/kg) was attenuated by approximately 50% in rats with MAP elevated through pretreatment with N(G)-nitro-L-arginine methyl ester (inhibitor of NO synthase), relative to that in rats with MAP elevated to a similar level through a continuous infusion of noradrenaline. Autonomic blockade with i.v. injections of mecamylamine (ganglion blocker) and propranolol (beta adrenoceptor antagonist) markedly augmented the depressor response to human urotensin II, but almost completely attenuated the tachycardia. The results suggest that the depressor response to human urotensin II is partially mediated via the NO/L-arginine pathway, and is suppressed by activity of the sympathetic nervous system. Furthermore, tachycardic response to human urotensin II is primarily mediated indirectly via baroreflex mechanisms. PMID- 12074942 TI - Protection against dextran sulfate sodium-induced colitis by microspheres of ellagic acid in rats. AB - Ellagic acid (EA), a naturally occurring plant phenol, has the antioxidant and anti-inflammatory activities. In the present study, we examined the effect of EA contained in microspheres on the ulcerative colitis induced experimentally in rats by dextran sulfate sodium (DSS). Experimental colitis was induced in male Fisher 344 rats by daily treatment with 3% DSS solution in drinking water for 7 days. EA of microspheres (mcEA: 1 approximately 10 mg/kg as EA contents) was administered p.o. twice daily for 6 days. In a preliminary study, we found that these microsphere capsules, when administered p.o., are effectively dissolved in the proximal to the ileo-cecal junction and distributed to the terminal ileum and the colon. The ulceration area, colon length, and mucosal myeloperoxidase (MPO) activity as well as thiobarbituric acid-reactive substances (TBARS) were measured on 7th day after the onset of DSS treatment. The DSS treatment for 7 days caused severe mucosal lesions in the colon, accompanied with the increases of MPO activity and TBARS as well as the decreases of body weight gain and colon length. Administration of mcEA reduced the severity of DSS-induced colitis in a dose dependent manner, and a significant effect was observed at 10 mg/kg, the ED50 being 2.3 mg/kg. This mcEA treatment also significantly mitigated changes in various biochemical parameters in the colonic mucosa induced by DSS. Although plain EA (without using microspheres) was also effective in reducing the severity of DSS-induced colitis, this effect was much less potent as compared with that of mcEA; the ED50 was about 15 times higher than that of mcEA. In addition, a significant effect on DSS-induced colitis was also obtained by intra-rectal administration of superoxide dismutase, an anti-oxidative agent. These results suggest that EA prevents the ulcerative colitis induced by DSS, probably by radical scavenging and/or anti-oxidative actions. The microspheres used in this study may be useful for delivering an orally administered drug specifically to the colon. PMID- 12074943 TI - The involvement of nitric oxide in the analgesic effects of ketamine. AB - We investigated the contribution of NO-cyclic GMP (cGMP) pathway to the antinociceptive effects of ketamine in mice by using the nitric oxide synthase inhibitor, nitro(g)- L-arginine methyl ester (L-NAME). Intraperitoneal (i.p.) (1, 5 or 10 mg/kg) or intrathecal (i.th.) (10, 30 or 60 microg/mouse) administration of ketamine produced dose-dependent antinociceptive effects in the acetic acid induced writhing and formalin tests but not in the tail-flick nor in hot-plate tests. Pretreatment of mice with L-NAME (10 mg/kg, i.p.) which produced no antinociception on its own, significantly inhibited the antinociceptive effect of ketamine (1, 5 or 10 mg/kg, i.p.). However, L-NAME (30 microg/mouse) was given intrathecally, it neither modified the antinociceptive effect of i.th. ketamine (10, 30 or 60 microg/mouse) nor did it produce an antinociceptive effect alone. These data suggest that the activation of the NO-cGMP pathway probably at the supraspinal level, but not spinal level, contributes to the antinociceptive effects of ketamine. PMID- 12074944 TI - Accommodation to simulations of defocus and chromatic aberration in the presence of chromatic misalignment. AB - Previous studies have demonstrated that accommodation will respond to sine gratings in which the relative modulations of red, green and blue image components have been altered to simulate the effects of defocus and longitudinal chromatic aberration. The present study aimed to determine the tolerance of the accommodative system to relative phase shifts in those components induced by chromatic misalignment. It was found that accommodation can tolerate moderate amounts of chromatic misalignment (6'), but responds adversely when misalignments are large. Applications to visual display terminals and spectacle lens and instrument design are discussed. PMID- 12074945 TI - Eye movements in the visual search of word lists. AB - The word identification span refers to the area of the visual field in which words can be identified during a single fixation. The purpose of the study was to estimate the vertical word identification span in a visual word search task, in which words were arranged in a vertical list. In addition, we studied the effect of list layout (orientation, length, and line spacing) on the speed of search and eye movements. The task of the observer was to identify a target word in a word list, where the other words were distracters. Threshold search time, that is, stimulus presentation time for correct identification at a probability level of 0.79, was determined by using a multiple alternative staircase method. Eye movements were recorded simultaneously. The results showed that, in vertical lists, 4-5 words could be identified during a single fixation. Thus, the vertical word identification span was 4-5 character spaces, whereas according to previous studies the horizontal word identification span is about 10 character spaces, which corresponds to 1-2 words. There were fewer fixations and the saccade amplitudes were smaller for vertical than for horizontal lists of the same length. However, search times did not depend on list orientation. This was due to longer fixation times for vertical lists. Further, since average fixation duration for vertical lists was longer than for horizontal lists, processing time seems to depend on the number of items within the span. PMID- 12074946 TI - The spatial distribution of selective attention assessed using the multifocal visual evoked potential. AB - To determine the manner in which attention is distributed among numerous locations in the visual space, we used a multifocal recording technique that allowed simultaneous recordings of evoked cortical activity from 12 visual field areas out to 23.6 degrees. We found that multifocal visual evoked potential (mfVEP) amplitude was larger when a region of visual space was attended than when it was not attended. The magnitude of this effect was inversely related to visual field eccentricity and there was no attention-related modulation of VEP amplitude for the most eccentric region. In addition, we found that mfVEP amplitudes in the regions contiguous to the attended region could also be larger, depending upon their spatial relationship to the attended region. Specifically, amplitudes in more central regions on the 'meridian of attention' were larger when the subject attended anywhere along that meridian. PMID- 12074947 TI - Visual coherence of moving and stationary image changes. AB - Detection thresholds were compared for moving and stationary oscillations with equivalent contrast changes. Motion was more detectable than stationary oscillation, and the difference increased with size of the feature (a Gaussian blob). Phase discriminations between a center and two flanking features were much better for motion than for stationary oscillation. Motion phase discriminations were similar to motion detection and were robust over increases in spatial separation and temporal frequency, but not so for stationary oscillations. Separate visual motion signals were positively correlated, but visual signals for stationary oscillation were negatively correlated. Evidently, motion produces visually coherent changes in image structure, but stationary contrast oscillation does not. PMID- 12074948 TI - Interactions between chromatic- and luminance-contrast-sensitive stereopsis mechanisms. AB - It is well known that chromatic information can assist in solving the stereo correspondence problem. It has also been suggested that there are two independent first-order stereopsis mechanisms, one sensitive to chromatic contrast and the other sensitive to luminance contrast (Vision Research 37 (1997) 1271). Could the effect of chromatic information on stereo correspondence be subserved by interactions between these mechanisms? To address this question, disparity thresholds (1/stereoacuity) were measured using 0.5 cpd Gabor patches. The stimuli possessed different relative amounts of chromatic and luminance contrast which could be correlated or anti-correlated between the eyes. Stereoscopic performance with these compound stimuli was compared to that with purely isoluminant and isochromatic stimuli at different contrasts. It was found that anti-correlated chromatic contrast severely disrupted stereopsis with achromatic stimuli and that anti-correlated luminance contrast severely disrupted stereopsis with chromatic stimuli. Less dramatic, but still significant, was the improvement in stereoacuity obtained using correlated colour and luminance contrast. These data are consistent with there being positive and negative interactions between chromatic and achromatic stereopsis mechanisms that take place after the initial encoding of disparity information, but before the extraction of stereoscopic depth. These interactions can be modelled satisfactorily assuming probability summation of depth sign information between independent mechanisms. PMID- 12074949 TI - Texture-orientation mechanisms pool colour and luminance contrast. AB - Do texture-sensitive mechanisms operate separately on, or pool, luminance and colour contrast information? We addressed this question by measuring threshold versus-amplitude functions for orientation-modulated (OM) gratings comprised of gabor elements defined by either colour or luminance contrast. In both the uncrossed (all elements in test and mask defined by either colour or luminance contrast) and crossed (equal mixtures of luminance and colour contrast in both test and mask) conditions, evidence of sub-threshold facilitation between test and mask was obtained. The sub-threshold facilitation in the crossed condition could not be accounted for by luminance artifacts in the ostensibly isoluminant gabors. The results are consistent with a single visual mechanism sensitive to OM textures that pools information from both the luminance and chromatic post receptoral mechanisms. PMID- 12074950 TI - Contrast sensitivity and the Stiles--Crawford effect. AB - We investigated the influence of the Stiles-Crawford effect (SCE) of the first kind on the contrast sensitivity function using the apodisation model of the SCE. The SCE was measured for the right eyes of two subjects using an increment threshold technique involving a two-channel Maxwellian-viewing system. Filters made of photographic film neutralised or doubled the SCE. Contrast sensitivities were measured with a 6 mm pupil diameter, defocus to +/-2D, and three SCE conditions (normal, neutralised and doubled). Modulation transfer functions were derived after measuring transverse aberrations with a vernier alignment technique, and were used to predict contrast sensitivity functions (CSFs). The measured CSFs were, in general, reasonable matches with the predicted CSFs. In particular, both demonstrated definite undulations ("notches") as defocus level increased. The influences of the SCE-modifying filters were generally of similar magnitude and direction to predictions, thus supporting the apodisation model of the SCE. The magnitudes of SCE influence between SCE-neutralised and SCE-doubled conditions were usually small at about 0.2-0.3 log unit, with a maximum influence of 0.5 log unit. Influences of the SCE were greater for myopic than for hypermetropic defocus. As measured by the CSF and an apodisation model, this study is in agreement with previous theoretical work and one experimental study in indicating that the SCE plays a minor role in improving spatial visual performance. PMID- 12074951 TI - Spatial-frequency properties of letter identification in amblyopia. AB - Amblyopia is characterized by a deficit in identifying small letters (acuity deficit) in the absence of identifiable ocular pathology. One explanation for this deficit is that the amblyopic visual system lacks appropriate channels tuned to high spatial frequencies for identifying small letters. The purpose of this study was to examine the spatial-frequency properties of letter identification in the amblyopic visual system. To do so, we measured contrast thresholds for identifying letters that were band-pass filtered to different bands of spatial frequencies, for letter sizes ranging from 2x to 19.2x larger than acuity letters. Letters were digitally filtered using a set of band-pass filters, with peak object spatial frequencies ranging from 0.88 to 10 c/letter. The bandwidth of the filters was 1 octave. For any given letter size, contrast sensitivity for identifying letters exhibits a spatial-tuning function. The shape of these tuning functions was found to be similar between amblyopic and non-amblyopic eyes, and across all letter sizes. The peak of these functions shifted progressively toward lower object spatial frequency when the letter size became smaller. When compared with the non-amblyopic eyes, the amblyopic eyes have a limited range of tuning functions sensitive to letters. However, when scaled with respect to acuity, the relationship between the peak frequency of the tuning functions and letter size becomes essentially identical in the amblyopic and non-amblyopic eyes. An ideal observer analysis that takes into account spectral information about letter identity and the contrast-sensitivity function of the observer, but does not invoke narrow-band channels, also shows that the properties of the tuning functions for letter identification are similar between the amblyopic and non amblyopic eyes. We conclude that the deficit in identifying small letters in amblyopes is not attributable to differences in the shape or selection of "channels", when compared with the normal visual system. Rather, it is a consequence of the difference in the resolution limit between the amblyopic and the non-amblyopic eyes. PMID- 12074952 TI - Left--right word recognition asymmetries in central and peripheral vision. AB - Word recognition for Western languages shows an increased probability of a correct response when words are presented to the right of fixation. We considered whether this right bias was consistent at eccentricities superior and inferior to fixation and whether this bias can be altered by different presentation strategies. A right bias of up to approximately 0.9 degrees to the right of fixation was found when words were presented along one horizontal meridian. The eccentricities tested extended up to 8 degrees above and below the point of fixation. However, the right bias was reduced for stimulus conditions where the word was randomly presented within a mosaic containing all possible presentation locations. We have therefore demonstrated that reading habit (right bias) can be manipulated based upon experimental paradigm, strongly supporting the proposition that the left-right asymmetry is a consequence of attending to a particular area of visual space as part of the normal reading habit, rather than an innate superiority for word recognition of the right visual field or reduced visual performance. PMID- 12074953 TI - A multi-layer sparse coding network learns contour coding from natural images. AB - An important approach in visual neuroscience considers how the function of the early visual system relates to the statistics of its natural input. Previous studies have shown how many basic properties of the primary visual cortex, such as the receptive fields of simple and complex cells and the spatial organization (topography) of the cells, can be understood as efficient coding of natural images. Here we extend the framework by considering how the responses of complex cells could be sparsely represented by a higher-order neural layer. This leads to contour coding and end-stopped receptive fields. In addition, contour integration could be interpreted as top-down inference in the presented model. PMID- 12074955 TI - Preface. PMID- 12074956 TI - Balancing the risks and benefits of moderate drinking. AB - The risks of excessive alcohol intake are well known, and heavy drinkers should decrease their intake or stop drinking. On the other hand, in comparison with non drinkers, moderate drinkers are at much lower risk of cardiovascular disease and certain other diseases and have lower total mortality. Thus, middle-aged or older men and post-menopausal women with no contraindications to alcohol use should be informed that they have, on average, net health benefits from the regular consumption of small-to-moderate amounts of alcohol. PMID- 12074957 TI - Alcohol and cardiovascular diseases: a historical overview. AB - Studying its history generally provides insights relevant to current understanding of a subject: the health effects of alcohol consumption is no exception to this rule. Perceiving past errors in the hopes of avoiding their repetition is crucial. Because there are clear disparities in the relationships of alcohol drinking to various cardiovascular conditions, attempts to simplify the subject of alcohol and cardiovascular diseases have delayed understanding this area. Thus, the following are considered separately: cardiomyopathy, arsenic and cobalt beer-drinkers' disease, cardiovascular beri-beri, systemic hypertension, cardiac arrhythmias, stroke, atherosclerotic coronary heart disease (CHD), total mortality, and definitions of safe drinking limits. The basic disparity underlying all alcohol-health relations is between effects of lighter and heavier drinking. PMID- 12074958 TI - Alcohol, type of alcohol, and all-cause and coronary heart disease mortality. AB - Many studies from a variety of countries have shown a U- or J-shaped relation between alcohol intake and mortality from all causes. It is now quite well documented from epidemiologic as well as clinical and experimental studies that the descending leg of the curve results from a decreased risk of cardiovascular disease among those with light-to-moderate alcohol consumption. The findings that wine drinkers are at a decreased risk of mortality from cardiovascular disease compared to non-wine drinkers suggest that substances present in wine are responsible for a beneficial effect on the outcome, in addition to that from a light intake of ethanol. Several potential confounding factors still remain to be excluded, however. PMID- 12074959 TI - Wine phenolics. AB - Wine contains many phenolic substances, most of which originate in the grape berry. The phenolics have a number of important functions in wine, affecting the tastes of bitterness and astringency, especially in red wine. Second, the color of red wine is caused by phenolics. Third, the phenolics are the key wine preservative and the basis of long aging. Lastly, since phenolics oxidize readily, they are the component that suffers owing to oxidation and the substance that turns brown in wine (and other foods) when exposed to air. Wine phenolics include the non-flavonoids: hydroxycinnamates, hydroxybenzoates and the stilbenes; plus the flavonoids: flavan-3-ols, the flavonols, and the anthocyanins. While polymeric condensed tannins and pigmented tannins constitute the majority of wine phenolics, their large size precludes absorption and thus they are not likely to have many health effects (except, perhaps, in the gut). The total amount of phenols found in a glass of red wine is on the order of 200 mg versus about 40 mg in a glass of white wine. PMID- 12074960 TI - Wine, biodiversity, technology, and antioxidants. AB - Two chemical classes of flavonoids, the flavan-3-ols (catechins and proanthocyanidins) and the anthocyanins, are the natural antioxidants present at the highest concentration in red grape and wine. In the berry, the anthocyanins are localized in the skins, similarly to other highly bioactive phenolics of grape such as the resveratrols and the flavonols, while the flavan-3-ols are contained both in the skins and seeds. During winemaking, only a fraction of the grape flavonoids are selectively extracted into the wine, with a time course and a final yield strongly depending on the grape variety. The knowledge of the diverse and cultivar-specific characteristics of the grape is therefore critical to the appropriate design of the winemaking process. By means of a selective extraction method specifically designed to mimic the winemaking process, it was possible to analyze the "phenolic potential of red grape," thus obtaining quantitative information about amount and localization of the extractable flavonoids in the grape. Twenty-five high-quality red grape cultivars (V. vinifera) were studied, including 4 of the worldwide leading cultivars and 21 Italian cultivars with the highest reputation for the production of both young and aged premium red wines. The results clearly indicate that the grape variety plays a central role in determining both the absolute amount of the flavonoids, and the distribution between the berry skin and seeds of the flavan-3-ols. The very high biodiversity of the red grape cultivars in terms of flavonoids indicate a largely under-exploited opportunity to produce a range of diverse premium wines with optimized levels of natural antioxidants. PMID- 12074961 TI - Chemistry of the antioxidant effect of polyphenols. AB - Most plant-derived polyphenols exhibit strong antioxidant potentials, established by various assay procedures. With pulse radiolysis experiments, absolute scavenging rate constants can be obtained with a variety of oxidizing radicals which allow further structure-activity correlations and, combined with EPR spectroscopy, detailed insight into the mechanisms governing these antioxidant reactions. The most striking difference occurs between regular flavonoids and both condensed and hydrolyzable tannins. The tannins are considered superior antioxidants as their eventual oxidation may lead to oligomerization via phenolic coupling and enlargement of the number of reactive sites, a reaction which has never been observed with the flavonoids themselves. PMID- 12074962 TI - Bioflavonoid-rich botanical extracts show antioxidant and gene regulatory activity. AB - Reactive oxygen and nitrogen metabolites are obligatory and essential products of metabolism. Unregulated increase in their production is associated with a number of chronic illnesses. Diets rich in fruits, vegetables, and wines are implicated in the prevention of chronic diseases. Molecular mechanisms by which fruits and vegetables confer their disease-preventive actions are poorly defined. However, recent developments in the fields of genomics and bioinformatics provide powerful tools to investigate the mechanisms by which botanicals affect cellular functions. This monograph illustrates the potential of large-scale messenger RNA analysis to unravel the role of transcription in mediating the effects of botanical extracts with antioxidant properties. The application of microarrays and oligonucleotide arrays shows multiple effects of antioxidant extracts on the expression of a broad spectrum of genes. PMID- 12074963 TI - Vasodilating procyanidins derived from grape seeds. AB - We have shown in previous work that extracts of grape seeds (GSE) and skins, grape juice, and many red wines exhibit endothelium-dependent relaxing (EDR) activity in vitro. This EDR activity involves endothelial nitric oxide (NO) release and subsequent increase in cyclic GMP levels in the vascular smooth muscle cells. The NO/cyclic GMP pathway is known to be involved in many cardiovascular-protective roles. The current study focuses on the isolation and identification of EDR-active compounds (procyanidins) from GSE. Crushed Concord grape seeds were extracted with methanol and the extract was separated into seven fractions (A-G) on a Toyopearl TSK-HW-40 column. EDR-active fractions (D-G) were further separated into 25 individual compound peaks by HPLC, 16 of which were EDR active (threshold for relaxation ranged between less than 0.5 microg/mL and greater than 4 microg/mL). Procyanidin identification was accomplished by electrospray-ion trap mass spectrometry (ES-ITMS), MS/MS, and by tannase treatment and acid thiolysis, followed by HPLC and ES-ITMS of the products. Activity of isolated procyanidins tended to increase with degree of polymerization, epicatechin content, and with galloylation. These EDR-active compounds (many of which also possess antioxidant activity), individually or in the form of wines, juices, or nutritional supplements, may be useful in preventing or treating cardiovascular diseases. PMID- 12074964 TI - Inhibition by wine polyphenols of peroxynitrite-initiated chemiluminescence and NADH oxidation. AB - Peroxynitrite (ONOO(-)) is a powerful oxidant produced by neutrophils, macrophages, and lymphocytes as a signaling and cytotoxic molecule from their primary production of nitric oxide (NO) and superoxide anion (O(2)(-)). In the vascular space, ONOO(-) will likely oxidize lipoproteins and promote atherogenesis. Pure wine flavonoids (catechin, epicatechin, myricetin), hydroxycinnamates (caffeic acid, ferulic acid, chlorogenic acid), and plain Argentine red wines were assayed as ONOO(-) scavengers in two assays: (a) ONOO(-) initiated chemiluminescence and (b) ONOO(-)-dependent oxidation. The assayed polyphenols as well as the red wines were effective inhibitors of the ONOO(-) driven oxidation reactions. Fifty percent of the pure substances were observed in the range of 30-300 microM and in the case of red wines with the equivalent of 80 120 microM of flavonoids. The amphipatic nature of wine polyphenols will lead to their accumulation at the lipoprotein surface, according to the Gibbs adsorption equation, where they are likely to prevent ONOO(-)-induced tyrosine nitration and LDL modification. PMID- 12074965 TI - The protective and anti-protective effects of ethanol in a myocardial infarct model. AB - Recent data indicate that acute alcohol exposure can have a preconditioning-like protective effect on the heart. We investigated the effect of ethanol exposure shortly before regional ischemia in an infarct model. Both in the open-chest rabbit and in the isolated rabbit heart, exposure of the heart to ethanol significantly reduced infarct size, but only if the alcohol were washed out or sufficiently metabolized before the onset of ischemia. If ethanol were still present during ischemia, it could not only prevent its own protective effect, but also abolish protection induced by ischemic preconditioning or the mitochondrial K(ATP) channel activator diazoxide. In the in vitro model, we tested for possible mediators of ethanol-induced protection and made comparisons to the signaling cascade of ischemic preconditioning. Neither adenosine receptor blockade with 8 (p-sulfophenyl) theophylline, scavenging of free radicals with N-2 mercaptopropionyl glycine, nor closure of K(ATP) channels with glibenclamide affected ethanol's protective effect. However, either a PKC inhibitor or a protein tyrosine kinase inhibitor could completely block ethanol-induced infarct size reduction. Both the protective and anti-protective effects of ethanol had a threshold of about 5 mM. Thus, ethanol-induced protection is mediated by protein kinase C and at least one protein tyrosine kinase, but, in contrast to ischemic preconditioning, is not triggered by either adenosine receptors, free radicals, or K(ATP) channels. Ethanol can only exert its protective effect if it is removed before the onset of ischemia. If still present during ischemia, ethanol has the opposite effect, and inhibits preconditioning by an as yet unidentified mechanism. PMID- 12074966 TI - Cardiovascular protection by alcohol and polyphenols: role of nitric oxide. AB - Cardiovascular disease, and in particular coronary heart disease (CHD), remains the leading cause of death in both men and women in the United States. Much epidemiologic evidence indicates that alcoholic beverages, and in particular red wine, results in a reduction in cardiovascular risk factors and decreases mortality; however, the mechanisms of this cardiovascular protection remains elusive. This review discusses evidence to suggest that *NO plays a critical role in cardiovascular protection and that nitric oxide synthase (NOS) is the responsible cardioprotective protein (see Bolli et al. 1998. Basic Res. Cardiol. 93: 325-338). PMID- 12074967 TI - Cardioprotection with alcohol: role of both alcohol and polyphenolic antioxidants. AB - Both epidemiological and experimental studies indicate that mild-to-moderate alcohol consumption is associated with a reduced incidence of mortality and morbidity from coronary heart disease. The consumption of wine, particularly red wine, imparts a greater benefit in the prevention of coronary heart disease than the consumption of other alcoholic beverages. The cardioprotective effects of red wine have been attributed to several polyphenolic antioxidants including resveratrol and proanthocyanidins. The results of our study documented that the polyphenolic antioxidants present in red wine, for example, resveratrol and proanthocyanidins, provide cardioprotection by their ability to function as in vivo antioxidants while its alcoholic component or alcohol by itself imparts cardioprotection by adapting the hearts to oxidative stress. Moderate alcohol consumption induced significant amount of oxidative stress to the hearts which was then translated into the induction of the expression of several cardioprotective oxidative stress-inducible proteins including heat shock protein (HSP) 70. Feeding the rats with red wine extract or its polyphenolic antioxidants as well as alcohol resulted in the improvement of postischemic ventricular function. Additionally, both wine and alcohol triggered a signal transduction cascade by reducing proapoptotic transcription factors and genes such as JNK-1 and c-Jun thereby potentiating an anti-death signal. This resulted in the reduction of myocardial infarct size and cardiomyocyte apoptosis. The results, thus, indicate that although both wine and alcohol alone reduce myocardial ischemic reperfusion injury, the mechanisms of cardioprotection differ from each other. PMID- 12074968 TI - Wine, diet, antioxidant defenses, and oxidative damage. AB - Oxidative stress is a central mechanism for the pathogenesis of ischemic heart disease and atherogenesis, for cancer and other chronic diseases in general, and it also plays a major role in the aging process. Dietary antioxidants constitute a large group of compounds that differ in mechanism of action, bioavailability and side effects. A systematic analysis of the role of the various antioxidants in chronic diseases is hampered by the difficulty of employing death or clinical events as end points in intervention studies. Therefore, valid markers for oxidative stress, which show dose response and are sensitive to changes in dietary supply of antioxidants, are potentially of great value when trying to establish healthy dietary patterns, or when one component, like red wine, is evaluated specifically. To evaluate potential oxidative stress markers we have studied the effect of different diets plus wine supplementation on antioxidant defenses and oxidative damage. In three experimental series, four groups of young male university students, one of older men and other of older women, 20-24 volunteers each, received Mediterranean or occidental (high-fat) diets alone or supplemented with red wine, white wine, or fruits and vegetables. Measurements included, leukocyte DNA 8-OH-deoxyguanosine (8OHdG), plasma 7 beta hydroxycholesterol, TBARS and well-characterized antioxidants, and plasma and urine polyphenol antioxidants. In all experimental groups that received red wine, consumption resulted in marked decrease in 8OHdG. The changes observed in 8OHdG correlate positively with the other markers of oxidative damage, and shows a clear inverse correlation with the plasma level of well established antioxidants and with measurements of total antioxidant capacity. Urinary total polyphenol content as well as the sum of some specific plasma species also correlate inversely with 8OHdG. In conclusion, the results identify 8OHdG as a very promising general marker of oxidative stress in nutrition intervention studies in humans, and red wine shows a remarkable protective effect. PMID- 12074969 TI - Wine flavonoids protect against LDL oxidation and atherosclerosis. AB - We have previously shown that consumption of red wine, but not of white wine, by healthy volunteers, resulted in the enrichment of their plasma LDL with flavonoid antioxidants such as quercetin, the potent free radicals scavenger flavanol, which binds to the LDL via a glycosidic ether bond. This phenomenon was associated with a significant three-fold reduction in copper ion-induced LDL oxidation. The ineffectiveness of flavonoid-poor white wine could be overcome by grape's skin contact for 18 hours in the presence of alcohol, which extracts grape's skin flavonoids. Recently, we observed that the high antioxidant potency of Israeli red wine could be related to an increased content of flavonols, which are very potent antioxidants and their biosynthesis is stimulated by sunlight exposure. To find out the effect (and mechanisms) of red wine consumption on atherosclerosis, we used the apo E deficient (E(0)) mice. In these mice, red wine consumption for two months resulted in a 40% decrement in basal LDL oxidation, a similar decrement in LDL oxidizability and aggregation, a 35% reduction in lesion size, and a marked attenuation in the number and morphology of lesion's macrophage foam cells. Red wine consumption resulted in accumulation of flavonoids in the mouse macrophages and these cells oxidized LDL and took up LDL about 40% less than macrophages from placebo-treated mice. Finally, the activity of serum paraoxonase (which can hydrolyze specific lipid peroxides in oxidized LDL and in atherosclerotic lesions) was significantly increased following consumption of red wine by E(0) mice. Red wine consumption thus acts against the accumulation of oxidized LDL in lesions as a first line of defense (by a direct inhibition of LDL oxidation), and as a second line of defense (by paraoxonase elevation and removal of atherogenic lesion's and lipoprotein's oxidized lipids). PMID- 12074970 TI - Scientific aspects that justify the benefits of the Mediterranean diet: mild-to moderate versus heavy drinking. AB - The Mediterranean diet is now recognized as being both limited in toxicity and abundant in nutrient and non-nutrient protective factors. A large body of basic, clinical and epidemiological studies have been developed in recent years to provide evidence of the benefits of the Mediterranean diet or its components on health. Evidence-based medicine ranks randomized controlled clinical trials as providing the highest level of evidence and expert opinions the lowest. On the basis of these criteria, the current state of knowledge about Mediterranean diet in primary and secondary prevention of disease and mortality and morbidity as functions of the amount of alcoholic beverage consumption, is reviewed. Efficacy versus effectiveness, the role of basic and animal research, and bioavailability studies providing evidence is also discussed. PMID- 12074971 TI - Antithrombotic effect of polyphenols in experimental models: a mechanism of reduced vascular risk by moderate wine consumption. AB - Epidemiological studies have suggested that cardiovascular disease can be decreased by moderate wine consumption, but an overall quantitative estimation of the relationship between wine intake and vascular risk is lacking. A meta analysis was therefore performed on 19 studies selected on the basis of the availability of specific information on the cardiovascular relative risk (RR) associated with wine consumption. A significant risk reduction (RR: 0.66, 95% CI 0.57-0.75) was associated with moderate (1-2 drinks or 150-300 mL/d) versus no wine consumption. In five studies which excluded ex-drinkers as reference group, the overall RR associated with wine consumption was 0.61 (95% CI 0.57-0.75). A dose-response relation between wine intake and vascular risk resulted in a J shaped curve, with a significant risk reduction at about 300 mL/d (trend analysis p = 0.032). Two studies were also performed to investigate the effects of wine polyphenols on experimental thrombosis in rats. Supplementation for 10 days with alcohol-free red wine--but not white wine or alcohol--induced a significant reduction of stasis-induced venous thrombosis, an effect blunted by NO synthase inhibitor L-NAME. In rats with diet-induced hyperlipidemia, alcohol-free red wine supplementation significantly delayed the thrombotic occlusion of an artificial prosthesis inserted into the abdominal aorta, but did not affect the increased cholesterol and triglyceride levels. TRAP values were significantly higher in animals receiving alcohol-free wine. Altogether these experimental data support an antithrombotic role of polyphenols in the reduced vascular risk associated with moderate wine consumption in man, as shown by our epidemiological studies. PMID- 12074972 TI - Thioredoxin superfamily and thioredoxin-inducing agents. AB - Mammalian thioredoxin (TRX) with redox-active dithiol in the active site plays multiple roles in intracellular signaling and resistance against oxidative stress. TRX is induced by a variety of stresses including infectious agents as well as hormones and chemicals. TRX is secreted from activated cells such as HTLV I-transformed T-cells as a redox-sensitive molecule with cytokine-like and chemokine-like activities. The promoter of the TRX gene contains a series of stress-responsive elements. In turn, TRX promotes activation of transcription factors such as NF-kappa B, AP-1, and p53. We have reported that natural substances including estrogen, prostaglandins, and cAMP induce mRNA, protein, and secretion of TRX. These agents seemed to exert their physiological functions including cytoprotective actions partly through the induction of TRX without massive oxidative stress, which induces TRX strongly as well as other stress proteins. We report here a new TRX inducer substance, geranylgeranylacetone (GGA), which is originally derived from a natural plant constituent and has been used in the clinical field as an anti-ulcer drug. We have demonstrated that GGA induces the messenger RNA and protein of TRX and affects the activation of transcription factors, AP-1 and NF-kappa B, and that GGA blunted ethanol-induced cytotoxicity of cultured hepatocytes and gastrointestine mucosal cells. We will discuss a possible novel molecular mechanism of GGA, which is to protect cells via the induction of TRX and activation of transcription factors such as NF-kappa B and AP-1. Identification of the particular TRX-inducing components may contribute to the elucidation of the molecular basis of the "French Paradox," in which good red wines are beneficial for the cardiovascular system. PMID- 12074973 TI - Wine polyphenols and optimal nutrition. AB - One of the key elements of Mediterranean diet is the use of wine, usually taken with foods. Besides the evidence from human experience and ancient medicine, modern experimental data support the notion that the most striking effect of wine in protecting against cardiovascular disease involves the reduction of oxidative damage to plasma lipoproteins. This oxidative damage is thought to be mediated by eating foods containing oxidized lipids. In fact, eating a meal containing oxidized lipids increases the plasma level of lipid hydroperoxides and increases the susceptibility to oxidation of LDL. The postprandial increase of LDL-, an oxidatively modified form of LDL, where apoB is unfolded and sinking in the core of the particle, is a valuable biomarker for this food-derived oxidative stress in plasma. Wine, taken with foods minimizes the postprandial rise of lipid hydroperoxides and LDL- and abolishes the increase of LDL oxidability. Among wine antioxidants, the best candidates for providing an antioxidant effect are procyanidins. These compounds are considered better antioxidants than the corresponding monomers containing catechol groups. This is due to the hydrogen transfer mechanism for the radical-scavenging reaction, which renders the reaction more specific for peroxyl radicals and pH independent. Moreover, the fast intramolecular disproportion among aroxyl radicals pulls the antioxidant reaction by both decreasing the oxidation potential and increasing the rate of the reaction. Apparently, wine procyanidins are active in preventing lipid oxidation of foods while in the digestive tract, thus preventing the postprandial plasma rise in oxidants. The likely limited bioavailability of these compounds, therefore, does not affect their relevance as key elements for optimizing nutrition and reducing risk of atherogenesis. Accordingly, studies with rabbits fed a high cholesterol diet show that grapeseed procyanidins are strongly protective not only in terms of reducing plasma lipid peroxides, but they also markedly inhibit lipid-laden foam-cell deposition. Drinking wine at meals provides this kind of protection, and the final benefits are realized by the prevention of the development of atheromatous lesions even under conditions of hypercholesterolemia. PMID- 12074974 TI - Cancer chemopreventive activity of resveratrol. AB - Cancer chemopreventive agents are designed to reduce the incidence of tumorigenesis by intervening at one or more stages of carcinogenesis. Recently, resveratrol, a natural product found in the diet of humans, has been shown to function as a cancer chemopreventive agent. Resveratrol was first shown to act as an antioxidant and antimutagenic agent, thus acting as an anti-initiation agent. Further evidence indicated that resveratrol selectively suppresses the transcriptional activation of cytochrome P-450 1A1 and inhibits the formation of carcinogen-induced preneoplastic lesions in a mouse mammary organ culture model. Resveratrol also inhibits the formation of 12-O-tetradecanoylphorbol-13-acetate (TPA)-promoted mouse skin tumors in the two-stage model. The enzymatic activities of COX-1 and -2 are inhibited by resveratrol in cell-free models, and COX-2 mRNA and TPA-induced activation of protein kinase C and AP-1-mediated gene expression are suppressed by resveratrol in mammary epithelial cells. In addition, resveratrol strongly inhibits nitric oxide generation and inducible nitric oxide synthase protein expression. NF kappa B is strongly linked to inflammatory and immune responses and is associated with oncogenesis in certain models of cancer, and resveratrol suppresses the induction of this transcription factor by a number of agents. The mechanism may involve decreasing the phosphorylation and degradation of I kappa B alpha. At the cellular level, resveratrol also induces apoptosis, cell cycle delay or a block in the G(1) --> S transition phase in a number of cell lines. Thus, resveratrol holds great promise for future development as a chemopreventive agent that may be useful for several disorders. Preclinical toxicity studies are underway that should be followed by human clinical trials. PMID- 12074975 TI - Resveratrol, a component of wine and grapes, in the prevention of kidney disease. AB - Ischemia is an inciting factor in 50% of incidences of acute renal failure, and it increases the risk of organ rejection after renal transplantation. We have previously demonstrated that resveratrol (RSV) reduces ischemia-reperfusion (I/R) injury of rat kidney both by antioxidant and anti-inflammatory mechanisms. However, a clear morphological demonstration of this activity has not been made. To answer this question we have performed a new set of experiments following the experimental protocol reported below to investigate the effects of I/R injury and RSV pretreatment on kidney morphology by computerized morphometric analysis. Both renal arteries were clamped for 40 minutes in 40 male Wistar rats (b.w. 220 +/- 20 g); 20 rats were pretreated with RSV 1 microM e.v. 40 minutes before clamping. All animals were reperfused for 24 hours and then sacrificed. Histological examination showed tissue conservation in treated rats. I/R-induced glomerular collapse (as revealed by mean glomerular volume and glomerular shape factor) was significantly reduced by RSV pretreatment. Capillary tuft/Bowman's capsule area ratio was enhanced in the I/R group suggesting tubular hypertension. RSV pre treatments significantly reduced this parameter to the control value. The number of platelet clots in the capillary tuft and tubular necrosis were also reduced by RSV versus I/R group. L-NAME administration worsened both functional and structural damage. Finally, cGMP urinary levels were markedly reduced from 12.1 +/- 8.4 nmol/day to 0.10 +/- 0.10 nmol/day in the I/R group. RSV provided cGMP (5.01 +/- 1.5 nmol/day, P < 0.05). As expected, L-NAME administration significantly reduced cGMP in urine (0.71 +/- 0.6 nmol/day). The present study confirms the protective effect of RSV pretreatment in I/R injury of rat kidney and suggests multiple mechanisms of action. PMID- 12074976 TI - Oxygen, oxidants, and antioxidants in wound healing: an emerging paradigm. AB - Disrupted vasculature and high energy-demand by regenerating tissue results in wound hypoxia. Wound repair may be facilitated by oxygen therapy. Evidence supporting the mode of action of hyperbaric oxygen in promoting wound healing is sketchy, however. Topical oxygen therapy involves local administration of pure oxygen. The advantages of topical oxygen therapy include low cost, the lack of systemic oxygen toxicity, and possibility of home treatment. While this modality of wound care is of outstanding interest, it clearly lacks the support of mechanism-oriented studies. The search for mechanisms by which oxygen supports wound healing has now taken another step. Respiratory burst-derived oxidants support healing. Oxidants serve as cellular messengers to promote healing. Although this information is of outstanding significance to the practice of oxygen therapy, it remains largely unexplored. The search for "natural remedies" has drawn attention to herbals. Proanthocyanidins or condensed tannins are a group of biologically active polyphenolic bioflavonoids that are synthesized by many plants. Proanthocyanidins and other tannins facilitate wound healing. A combination of grape seed proanthocyanidin extract and resveratrol facilitates inducible VEGF expression, a key element supporting wound angiogenesis. Strategies to manipulate the redox environment in the wound are likely to be of outstanding significance in wound healing. PMID- 12074977 TI - Protective effects of a novel niacin-bound chromium complex and a grape seed proanthocyanidin extract on advancing age and various aspects of syndrome X. AB - Aging is the progressive accumulation of changes with time that are responsible for the ever-increasing likelihood of disease and death. The precise cascade of pathological events mainly responsible for aging are still not clearly understood, but enhanced production of free radicals and its deleterious effects on proteins, nucleic acids, and fats, as well as enhanced glycosylation of proteins and DNA are prevalent during aging. Insulin resistance may be a common etiology, at least in part, behind the pathobiological alterations of advancing age. Prevalent age-related disorders such as cardiovascular diseases, obesity, and cancer have been associated with impaired glucose/insulin metabolism and its consequences. This leads to future strategies to combat the aging process and chronic disorders such as the components of syndrome X associated with aging. Increasing the intake of antioxidants and/or substances recognized to enhance insulin sensitivity is a natural means of combatting the glucose/insulin perturbations and free radical damage. Accordingly, ingestion of niacin-bound chromium and natural antioxidants such as grape seed proanthocyanidin extract has been demonstrated to improve insulin sensitivity and/or ameliorate free radical formation and reduce the signs/symptoms of chronic age-related disorders including syndrome X. These natural strategies possess a highly favorable risk/benefit ratio. PMID- 12074978 TI - Cellular protection with proanthocyanidins derived from grape seeds. AB - Grape seed proanthocyanidins have been reported to possess a broad spectrum of pharmacological and medicinal properties against oxidative stress. We have demonstrated that IH636 proanthocyanidin extract (GSPE) provides excellent protection against free radicals in both in vitro and in vivo models. GSPE had significantly better free radical scavenging ability than vitamins C, E and beta carotene and demonstrated significant cytotoxicity towards human breast, lung and gastric adenocarcinoma cells, while enhancing the growth and viability of normal cells. GSPE protected against tobacco-induced apoptotic cell death in human oral keratinocytes and provided protection against cancer chemotherapeutic drug induced cytotoxicity in human liver cells by modulating cell cycle/apoptosis regulatory genes such as bcl2, p53 and c-myc. Recently, the bioavailability and mechanistic pathways of cytoprotection by GSPE were examined on acetaminophen induced hepatotoxicity and nephrotoxicity, amiodarone-induced pulmonary toxicity, doxorubicin-induced cardiotoxicity, DMN-induced immunotoxicity and MOCAP-induced neurotoxicity in mice. Serum chemistry changes, integrity of genomic DNA and histopathology were assessed. GSPE pre-exposure provided near complete protection in terms of serum chemistry changes and DNA damage, as well as abolished apoptotic and necrotic cell death in all tissues. Histopathological examination reconfirmed these findings. GSPE demonstrated concentration-/dose-dependent inhibitory effects on the drug metabolizing enzyme cytochrome P450 2E1, and this may be a major pathway for the anti-toxic potential exerted by GSPE. Furthermore, GSPE treatment significantly decreased TNFalpha-induced adherence of T-cells to HUVEC by inhibiting VCAM-1 expression. These results demonstrate that GSPE is highly bioavailable and may serve as a potential therapeutic tool in protecting multiple target organs from structurally diverse drug- and chemical-induced toxicity. PMID- 12074979 TI - Polyphenols and red wine as peroxynitrite scavengers: a chemiluminescent assay. AB - A novel chemiluminescent assay for evaluating peroxynitrite (ONOO(-))-scavenging capacity was developed. The experimental protocol ensures sensitivity and reproducibility of measurements. The addition of 0-500 microM ONOO(-) to rat liver homogenate generated a luminous signal that was analyzed by chemiluminescence in a LKB Wallac liquid scintillation counter. The obtained optimal conditions were: 1-2 mg/mL of homogenate protein in 120 mM KCl, 30 mM phosphate buffer (pH 7.4), and 220 microM ONOO(-) at 30 degrees C. As polyphenols we used (+)-catechin, (-)-epicatechin, and myricetin. The most efficient of the compounds tested was myricetin with an IC(50) of 20 microM. The effectiveness of this method was verified by evaluating the antioxidant ability of three red wine samples to decrease peroxynitrite-initiated chemiluminescence. The ONOO(-) scavenging activity of wines measured by this assay was related to the phenolic level of the samples. The quickness and reliability of this assay makes it particularly suitable for a large-scale screening of watery food extracts. PMID- 12074980 TI - Polyphenols in red wines prevent NADH oxidation induced by peroxynitrite. AB - It is known that red wines have antioxidant properties owing to the presence of polyphenols and hydroxycinnamates. It has been reported that these compounds are efficient scavengers of peroxynitrite (ONOO(-)). We assayed the ONOO(-) scavenging activity of plant polyphenols, hydroxycinnamates, and red wines using a fluormetric assay that involves the participation of ONOO(-) (200 microM) as oxidant and NADH (50-100 microM) as a target molecule. NADH oxidation was prevented by pure polyphenols and hydroxycinnamates. With the wines, the Cabernet Sauvignon showed more potent ONOO(-)-scavenging ability than the Malbecs and the blended wines. These results correlate well with the total phenol content determined by Folin-Ciocalteau assay. PMID- 12074981 TI - Comparative study on the antioxidant capacity of wines and other plant-derived beverages. AB - Consistent epidemiological data point to a reduced morbidity and mortality from coronary heart disease and atherosclerosis in people consuming plant-derived beverages such as tea or wine. We studied the antioxidant capacity of three red wines (W) and compared it those of tea and herbal "mate" tea infusions. The antioxidant capacity was evaluated measuring: (1) the inhibition of the luminol induced chemiluminescence assay (TRAP); (2) the inhibition of 2.2'-thiobarbituric reactive substances (TBARS) formation in liposomes by fluorescence; (3) the protection of Jurkat cells from AMVN-induced oxidation, measuring the oxidation of 5-(and-6)-carboxy-2'7'-dichlorodihydrofluorescein diacetate to a fluorescent derivative. The polyphenolic content was estimated spectrophotometrically and by HPLC with electrochemical detection. All three beverages provided antioxidant protection in the three assays in a dose-dependent manner. Significant and positive correlations were found between antioxidant capacity and total polyphenol content, especially in the Jurkat cell oxidation assay (r: 0.96, p < 0.01). Results suggest that these dietary components could be a source of antioxidants that protect from oxidative stress. Further studies of absorption and metabolism of the active compounds will judge the physiological relevance of these results for human health. PMID- 12074982 TI - Assessing the antioxidant capacity in the hydrophilic and lipophilic domains: study of a sample of Argentine wines. AB - The antioxidant capacity of different types of red and white wines was assessed by different assays. Two assays were used to evaluate the antioxidant capacity in aqueous phase: (1) inhibition of the generation of 2,2'-azinodi(3 etilbencenotiazolin-6-sulfonate) (ABTS)-derived radical; and (2) protection of vitamin E in human plasma. The results indicated that red wines were not all equally effective, and a comparison could be made among different types. The ABTS assay showed that Cabernet Sauvignon wines had the highest antioxidant capacity followed by Malbec wines. Red wines showed a protective capacity of 70-90% in preventing SH-groups oxidation, and, in addition, they were also effective in preventing the oxidative damage in lipid domains (30-97% protection to liposomes, 20-70% to vitamin E). The antioxidant capacity of the white wines was significantly lower, as evaluated by all the assays. Significant correlations were found for phenolics and catechin content with the antioxidant capacity of the studied wines. PMID- 12074983 TI - Physiological model of the stimulative effects of alcohol in low-to-moderate doses. AB - A physiological model of alcohol use is proposed, based on the observation that ethanol is a natural intracellular metabolite. It is suggested that ethanol synthesis is an intermediate step in the metabolic pathway of elimination of the excess of energy-releasing substrates from mitochondria and the cytosol when the cell does not require additional energy. The process is first of all important for elimination of acetyl-CoA from mitochondria. Ingested alcohol passes across plasma and mitochondrial membranes by diffusion and follows the endogenous ethanol pathway but in the reverse direction. This pathway is used when the cell needs additional energy. In that case, exogenous ethanol supplies the respiratory chain with electrons via NADH on the path of oxidation to acetaldehyde and then to acetate by alcohol and aldehyde dehydrogenases. After conversion to acetyl CoA, it goes into the citrate cycle and is thus used for energy production entirely. If citrate cycle activity is decreased constitutionally, acetyl-CoA enters into citrate cycle too slowly and energy deficit arises, ethanol compensates for the energy deficit. Acetate then diffuses out of mitochondria. The less energy production in mitochondria, the more the cell is ready to accept alcohol as an energy source. Citrate cycle activity can be depressed as a result of single-nucleotide polymorphisms of genes controlling some enzymes involved in the cycle, which influences the alcohol requirement. PMID- 12074984 TI - A queue paradigm formulation for the effect of large-volume alcohol intake on the lower urinary tract. AB - We examine the urologic consequences of large-volume alcohol intake in light of recent advances in hollow viscera biomechanics and urinary tract pharmacology. Recent studies have shown that alcohol is depressive on the isolated rabbit detrusor. Patients teetering on or having manifest pelvi-ureteric junction obstruction or bladder outflow obstruction have been known to demonstrate symptoms on alcohol consumption. In men over 50, the cause is usually an enlarged prostate. Loin pain is precipitated in the case of the former and urinary retention in the latter case after consuming alcohol. Causation is difficult to prove for ethical reasons as well as the practical difficulties in running a prospective trial. It appears that a combination of rapid fill (due to diuretic effects), a weakened pump (due to the depressive effect of alcohol at least with the detrusor) and outlet obstruction lead to the loin pain and urinary retention seen in clinical practice. People liable to urinary tract obstruction would be well advised to avoid large volume alcohol intakes. PMID- 12074985 TI - Oxidative stress and inflammatory reaction modulation by white wine. AB - Wine and olive oil, essential components of the Mediterranean diet, are considered important factors for a healthy life style. Tyrosol (T) and caffeic acid (CA) are found in both extra virgin olive oil and in white wine. Three white wines from the northeast Italy and four white wines from Germany were analyzed for their content of T and CA. These compounds were tested for their antioxidant activity and their capacity to modulate three different cytokines: IL-1 beta, IL 6, and TNF-alpha, which are currently considered to be the major cytokines influencing the acute phase of the inflammatory response. Furthermore, the antioxidant activity of T and CA was analyzed by monitoring the oxidation of a redox-sensitive probe by using laser scanning confocal microscopy. T and CA, applied at nanomolar range, were found to significantly reduce the generation of oxidants induced by azobis-amidinopropanedihydrochloride. Peripheral blood mononuclear cells (PBMC) from healthy volunteers were incubated at 37 degrees C for 12 hours with 100 ng LPS (E. coli and P. maltofilia). Increasing doses of T and CA (150 nM to 300 microM) were added and cell-associated IL-1 beta and TNF alpha were determined by immunoreactive tests after three freeze-thaw cycles. IL 6 release was also determined in cell surnatants. LPS-stimulated PBMC showed a significant increase in cytokine release, while T and CA, used at nanomolar concentrations, were able to modulate their expression. Taken together, these results suggest a remarkable effect of white wine non-alcoholic compounds on oxidative stress and inflammatory reaction. PMID- 12074986 TI - Reduction of myocardial ischemia reperfusion injury with regular consumption of grapes. AB - Recently several polyphenolic antioxidants derived from grape seeds and skins have been implicated in cardioprotection. This study was undertaken to determine if the grapes were equally cardioprotective. Sprague Dawley male rats were given (orally) standardized grape extract (SGE) for a period of three weeks. Time matched control experiments were performed by feeding the animals 45 microg/100 of glucose plus 45 microg/100 g fructose per day for three weeks. After 30 days, rats were sacrificed, hearts excised and perfused via working-mode. Hearts were made ischemic for 30 min followed by two hours of reperfusion. At 100 mg/kg and at 200 mg/kg, SGE provided significant cardioprotection as evidenced by improved post-ischemic ventricular recovery and reduced amount of myocardial infarction. No cardioprotection was apparent when rats were given grape samples at a dose of 50 mg/100 g/day. In vitro studies demonstrated that the SGE could directly scavenge superoxide and hydroxyl radicals which are formed in the ischemic reperfused myocardium. The results demonstrate that the heats of the rats fed SGE reduced myocardial ischemia reperfusion injury by functioning as in vivo antioxidant. PMID- 12074988 TI - Doctor, should I have a drink? An algorithm for health professionals. PMID- 12074987 TI - Cardioprotective abilities of white wine. AB - To study if white wines, like red wine, can also protect the heart from ischemia reperfusion injury, ethanol-free extracts of three different white wines (WW1, WW2 and WW3) (100 mg/100 g body weight) were given orally to Sprague Dawley rats (200 g body weight) for three weeks. Control rats were given water only for the same period of time. After three weeks, rats were anesthetized and sacrificed, and the hearts excised for the preparation of isolated working rat heart. All hearts were subjected to 30 min global ischemia followed by two hours of reperfusion. The results demonstrated that among the three different white wines, only WW2 showed cardioprotection as evidenced by improved post-ischemic ventricular recovery compared to control. The amount of malonaldehyde production in white wine-fed rat hearts were lower compared to that found in control hearts indicating reduced formation of the reactive oxygen species. In vitro studies using chemiluminescence technique revealed that these white wines scavenged both superoxide anions and hydroxyl radicals. The results of our study demonstrated that only WW2 white wine provided cardioprotection as evidenced by the improved the post-ischemic contractile recovery and reduced myocardial infarct size. The cardioprotective effect of this white wine may be attributed, at least in part, from its ability to function as an in vivo antioxidant. PMID- 12074989 TI - Preservation of paraoxonase activity by wine flavonoids: possible role in protection of LDL from lipid peroxidation. AB - Paraoxonase is an esterase physically associated with HDL, and its activity has been shown to be inversely related to the risk of cardiovascular diseases. We have shown that paraoxonase can hydrolyze specific lipid peroxides in oxidized lipoproteins and in atherosclerotic lesions. Paroxonase was shown to be inactivated by oxidative stress. Consumption of wine flavonoids was shown to preserve paraoxonase activity by reducing the oxidative stress in apolipoprotein E-deficient mice, thereby contributing to paraoxonase hydrolytic activity on lipid peroxides in oxidized lipoproteins and atherosclerotic lesions. PMID- 12074990 TI - Absorption and metabolism of antioxidative polyphenolic compounds in red wine. AB - We have shown that drinking red wine reduces oxidation of LDL. This reduction in oxidation has been attributed to the polyphenolic compounds in red wine, but the mechanisms of absorption and metabolism of these compounds has been unclear. We therefore investigated the absorption and metabolism of polyphenols using rats to identify their active forms in biological fluids. We also investigated the effect of tartaric acid (TA), a major organic acid in wine, on the absorption of polyphenols. Our results suggested that low molecular weight polyphenols are absorbed in the intestine and metabolized to their glucuronide conjugates, which exhibit antioxidative activity in plasma, and that TA can enhance the bioavailability of wine polyphenols. PMID- 12074991 TI - The in vivo antithrombotic effect of wine consumption on human blood platelets and hemostatic factors. AB - We compared the in vivo effect of red vs. white wine consumption on platelet aggregation, responsiveness and membrane viscosity, plasma total antioxidant status, thromboxane B(2) levels, and fibrinolysis. Diet and red wine had a synergistic effect in decreasing platelet aggregation. Red wine did not have a significantly more favorable effect on the fibrinolytic factors than white wine. The reduction in platelet membrane viscosity after red wine, which could contribute to the protective antithrombotic role of red wine, needs further explanation. PMID- 12074992 TI - Corrupt captains and convicts. AB - Australia is unique in that so many of its wine companies were founded by members of the medical profession. The First Fleet, which brought the first convicts and settlers to Australia in 1787, was delayed until wine was provided as a medicine for the long voyage from England. Australia's wine doctors were advocating the use of wine as a medicine 200 years before the French Paradox. PMID- 12074993 TI - The cardioprotective effect of wine on human blood chemistry. AB - We investigated the in vivo effects of regular consumption of red and white wine on the serum lipid profile, plasma plasminogen activator-1, homocysteine levels, and total antioxidant status. This study confirmed that moderate consumption of wine, red more than white, exerts cardioprotective effects through beneficial changes in lipid profiles and plasma total antioxidant status. PMID- 12074994 TI - Melatonin: an antioxidant in edible plants. AB - Melatonin, a molecule with antioxidant properties that is widely distributed in the animal kingdom, has now been shown to exist in the plant kingdom, including edible plants. Our findings show that melatonin is not only an endogenously produced antioxidant, but that it is also consumed in the diet. Since melatonin concentrations in the blood correlate with the total antioxidant status of this fluid, it is likely that dietary melatonin could be important in protecting against oxidative damage. PMID- 12074995 TI - EPR, free radicals, wine, and the industry: some achievements. AB - The achievements and uses of electron paramagnetic resonance in the wine industry are pointed out. For example, it first detected stable free radicals in red and white wines and showed that the radical concentration, and therefore the antioxidant action, of white wines was increased by skin and oak exposure. It is expected that EPR will be used more in the future. PMID- 12074996 TI - For each a road. PMID- 12074997 TI - The hippocampus and memory of verbal and pictorial material. AB - Recognition of words and kaleidoscope pictures showed a double dissociation of left and right hippocampal activity using magnetic source imaging (MSI). MSI has advantages over alternative imaging techniques that measure hemodynamic changes for identifying regional changes in brain activity in real time and on an individual subject basis without the need for image subtraction. In this study, lists of words or kaleidoscope pictures were presented for memorization followed by tests of list items and foils during which brain activity was recorded. There was greater activation in the left than the right hippocampus with abstract nouns (e.g., relief) and greater activation in the right than the left hippocampus with kaleidoscope pictures. This dissociation was evident on a case by case basis. This study demonstrates the specialization of the two medial temporal lobe (MTL) regions, including the hippocampi, for mnemonic processing of verbal and pictorial items that are difficult to encode verbally. PMID- 12074998 TI - Mapping of olfactory memory circuits: region-specific c-fos activation after odor reward associative learning or after its retrieval. AB - Although there is growing knowledge about intracellular mechanisms underlying neuronal plasticity and memory consolidation and reconsolidation after retrieval, information concerning the interaction among brain areas during formation and retrieval of memory is relatively sparse and fragmented. Addressing this question requires simultaneous monitoring of activity in multiple brain regions during learning, the post-acquisition consolidation period, and retrieval and subsequent reconsolidation. Immunoreaction to the immediate early gene c-fos is a powerful tool to mark neuronal activation of specific populations of neurons. Using this method, we are able to report, for the first time, post-training activation of a network of closely related brain regions, particularly in the frontal cortex and the basolateral amygdala (BLA), that is specific to the learning of an odor reward association. On the other hand, retrieval of a well-established associative memory trace does not seem to differentially activate the same regions. The amygdala, in particular, is not engaged after retrieval, whereas the lateral habenula (LHab) shows strong activation that is restricted to animals having previously learned the association. Although intracellular mechanisms may be similar during consolidation and reconsolidation, this study indicates that different brain circuits are involved in the two processes, at least with respect to a rapidly learned olfactory task. PMID- 12074999 TI - The effect of similarity between elemental stimuli and compounds in olfactory patterning discriminations. AB - We studied the ability of honeybees to discriminate between single odorants and binary olfactory mixtures. We analyzed the effect of the number of common elements between these two stimulus classes on olfactory discrimination. We used olfactory conditioning of the honeybees' proboscis extension reflex (PER), a paradigm in which odors can be associated with a reinforcement of sucrose solution. Bees were asked to discriminate reinforced from nonreinforced olfactory stimuli. They were trained with two elemental odors (A and B) versus a binary olfactory mixture. The mixture was either AB (group 2CE, two common elements), AC (group 1CE, one common element A), or CD (group 0CE, no common element). Three groups followed a positive patterning schedule (mixture reinforced and elements nonreinforced: groups 2CE+, 1CE+, and 0CE+) and three other groups a negative patterning schedule (mixture nonreinforced and elements reinforced: groups 2CE-, 1CE-, and 0CE-). We showed that a reduction of similarity (number of common elements) between elemental odors and compounds enhanced the ability to discriminate elements from compounds and that the kind of compound processing used by the bees supports theories that assume nonelemental compound processing (i.e., that exclude the mere summation of the elemental associative strengths upon compound presentation). PMID- 12075000 TI - Successive olfactory reversal learning in honeybees. AB - Honeybees Apis mellifera can associate an originally neutral odor with a reinforcement of sucrose solution. Forward pairings of odor and reinforcement enable the odor to release the proboscis extension reflex in consecutive tests. Bees can also be conditioned differentially: They learn to respond to a reinforced odor and not to a nonreinforced one. They can also learn to reverse their choice. Here we ask whether honeybees can learn successive olfactory differential conditioning tasks involving different overlapping pairs of odors. The conditioning schedules were established in order to train the animals with 3, 2, 1, or 0 reversals previous to a last differential conditioning phase in which two additional reversals were present. We studied whether or not successive reversal learning is possible and whether or not learning olfactory discrimination reversals affects the solving of subsequent discrimination reversals. Therefore we compared the responses of bees that had experienced reversals with those of bees that had not experienced such reversals when both are confronted with a new reversal situation. In experiment 1 we showed that bees that had experienced three previous reversals were better in solving the final reversal task than bees with no previous reversal experience. In experiment 2, we showed that one reversal learning is enough for bees to perform better in the final reversal task. The successive different reversals trained in our experiments resemble the natural foraging situation in which a honeybee forager has to switch successively from an initial floral species to different ones. The fact that experiencing such changes seems to improve a bee's performance in dealing with further new exploited food sources has therefore an adaptive impact for the individual and for the colony as a whole. PMID- 12075001 TI - A new group-training procedure for habituation demonstrates that presynaptic glutamate release contributes to long-term memory in Caenorhabditis elegans. AB - In the experiments reported here we have developed a new group-training protocol for assessing long-term memory for habituation in Caenorhabditis elegans. We have replicated all of the major findings of the original single-worm protocol using the new protocol: (1) distributed training produced long-term retention of training, massed training did not; (2) distributed training at long interstimulus intervals (ISIs) produced long-term retention, short ISIs did not; and (3) long term memory for distributed training is protein synthesis-dependent as it could be blocked by heat shock during the inter-block interval. In addition, we have shown that long-term memory for habituation is graded, depending on the number of blocks of stimuli in training. The inter-block interval must be >40 min for long term retention of training to occur. Finally, we have tested long-term memory for habituation training in a strain of worms with a mutation in a vesicular glutamate transporter in the sensory neurons that transduce tap (eat-4). The results from these eat-4 worms indicate that glutamate release from the sensory neurons has an important role in the formation of long-term memory for habituation. PMID- 12075002 TI - The time course of neural changes underlying auditory perceptual learning. AB - Improvement in perception takes place within the training session and from one session to the next. The present study aims at determining the time course of perceptual learning as revealed by changes in auditory event-related potentials (ERPs) reflecting preattentive processes. Subjects were trained to discriminate two complex auditory patterns in a single session. ERPs were recorded just before and after training, while subjects read a book and ignored stimulation. ERPs showed a negative wave called mismatch negativity (MMN)-which indexes automatic detection of a change in a homogeneous auditory sequence-just after subjects learned to consciously discriminate the two patterns. ERPs were recorded again 12, 24, 36, and 48 h later, just before testing performance on the discrimination task. Additional behavioral and neurophysiological changes were found several hours after the training session: an enhanced P2 at 24 h followed by shorter reaction times, and an enhanced MMN at 36 h. These results indicate that gains in performance on the discrimination of two complex auditory patterns are accompanied by different learning-dependent neurophysiological events evolving within different time frames, supporting the hypothesis that fast and slow neural changes underlie the acquisition of improved perception. PMID- 12075003 TI - Genetic susceptibility to childhood common acute lymphoblastic leukaemia is associated with polymorphic peptide-binding pocket profiles in HLA-DPB1*0201. AB - In a previous study, we obtained preliminary evidence in a small series of patients (n = 63) suggesting that susceptibility to childhood common acute lymphoblastic leukaemia (c-ALL) was associated with an allele at the HLA-DPB1 locus, DPB1*0201. We have now tested this hypothesis by comparing the frequency of children with leukaemia (n = 982) who typed for specific DPB1 alleles and two groups of non-leukaemic children, one consisting of children with solid tumours, excluding lymphomas (n = 409), the other consisting of normal infants (n = 864). We found that significantly more children with c-ALL and T-ALL, but not pro-B ALL or acute non-ALL typed for DPB1*0201 as compared with children with solid tumours [odds ratio (OR), 95% confidence interval (CI) for c-ALL: 1.76, 1.20-2.56; T-ALL: 1.93, 1.01-3.80] and normal infants (OR, 95% CI for c-ALL: 1.83, 1.34-2.48; T ALL: 2.00, 1.10-3.82). In childhood c-ALL, significantly more children than those with solid tumours or normal infants typed for DPB1 alleles coding specific polymorphic amino acids lining the antigen-binding site of the DPbeta1*0201 allotypic protein, suggesting that susceptibility to childhood c-ALL may be influenced by DPbeta ABS amino acid polymorphisms shared by DPbeta1*0201 and other DPbeta1 allotypes. These results point to a mechanism of c-ALL susceptibility that involves the presentation of specific antigenic peptides, possibly derived from infectious agents, by DPbeta1*0201-related allotypic proteins, leading to the activation of helper T cells mediating proliferative stress on preleukaemic cells. PMID- 12075004 TI - Fine mapping of a putative tuberculosis-susceptibility locus on chromosome 15q11 13 in African families. AB - Host genetics plays an important role in individual susceptibility and resistance to infectious diseases, but no genes have yet been identified using genome-wide screens. Twin studies have indicated that tuberculosis susceptibility has a significant host genetic component, and several genes appear to be involved. Recently, a genome-wide linkage analysis of 136 African families identified chromosome 15q11-13 as a region with suggestive evidence of linkage, with a LOD score of 2.0. We tested 10 microsatellite markers and 5 positional candidate genes in this chromosomal region for deviation from random transmission from parents to affected offspring. The polymorphisms, lying in a region of 14 cM, were initially typed in the same 79 Gambian families used in the genome screen. A borderline significant association with a 7 bp deletion in UBE3A (P = 0.01) was found. This polymorphism was then evaluated further in a larger series of families with tuberculosis, including 44 Guinean families and 57 families from South Africa. Testing for association between the deletion and tuberculosis across all the families using the exact symmetry test further supported the association (overall P = 0.002). These fine-mapping data suggest that UBE3A or a closely flanking gene may be a tuberculosis-susceptibility locus. PMID- 12075005 TI - Survival motor neuron (SMN) protein: role in neurite outgrowth and neuromuscular maturation during neuronal differentiation and development. AB - Childhood spinal muscular atrophy (SMA) is a common neuromuscular disorder caused by absent or deficient full-length survival motor neuron (SMN) protein. Clinical studies and animal models suggest that SMA is a developmental defect in neuromuscular interaction; however, the role of SMN in this process remains unclear. In the present study, we have determined the subcellular localization of SMN during retinoic-acid-induced neuronal differentiation of mouse embryonal teratocarcinoma P19 cells as well as in skeletal muscle during the critical period of neuromuscular maturation. We demonstrate, for the first time, SMN accumulation in growth-cone- and filopodia-like structures in both neuronal- and glial-like cells, identifying SMN as a new growth cone marker. Indeed, SMN was present at the leading edge of neurite outgrowths, suggesting that SMN may play a role in this process. In addition, SMN was detected as small dot-like particles within the cytoplasm of skeletal muscle during the first 2 weeks after birth, but their number peaked by P6. Intense SMN staining in neuromuscular junctions was observed throughout the entire postnatal period examined. Taken together, these results suggest that SMN may indeed fulfill neuronal- and muscle-specific functions, providing a more plausible mechanism explaining motor neuron degeneration and associated denervation atrophy of skeletal muscles in SMA. The primary SMA pathology most likely initiates in the peripheral axon--the result of deficient neurite outgrowth and/or neuromuscular maturation. PMID- 12075006 TI - In vivo electrotransfer of the cardiotrophin-1 gene into skeletal muscle slows down progression of motor neuron degeneration in pmn mice. AB - Among all vectors designed for gene therapy purposes, adenovirus appears to be the most efficient in vivo vehicle to transduce the broadest spectrum of cellular targets. However, the deleterious immunogenicity of this viral vector impedes its use in chronic diseases. Non-viral vectors, such as naked DNA, are attractive alternatives for safety and technical issues, such as scale-up production. Naked DNA injection, greatly improved when combined with electroporation, showed great potential in adult animals, especially when directed to the muscle. We have recently proven the therapeutic effect of a neonatal single intramuscular injection of a cardiotrophin-1 (CT-1)-encoding adenovirus in a hereditary disease mouse model of human motor neuron disease, the progressive motor neuronopathy (pmn) mutant. We now demonstrate that a single injection/electroporation of a CT 1-encoding plasmid in neonate pmn mice is almost as efficient as adenovirus mediated gene transfer with respect to survival, muscular function and neuroprotection of the animals. Treated mice gain global weight, their mean lifespan is extended by 25%, all their electromyographic parameters are improved and myelinated axons of their phrenic nerves are protected. Moreover, we show that re-injection/electroporation leads to improvements in this neuroprotection. We therefore demonstrate for the first time the therapeutic efficacy of neonatal intramuscular DNA injection/electroporation in a murine model of a human hereditary disorder. PMID- 12075007 TI - An evolutionarily conserved putative insulator element near the 3' boundary of the imprinted Igf2/H19 domain. AB - Igf2 and H19 are closely linked imprinted genes lying at the centromeric end of a 1 Mb imprinted domain on mouse chromosome 7. L23mrp and other genes located 3' (more centromeric) to H19 are not imprinted and do not interact with the enhancers shared by Igf2 and H19. It is therefore suggested that the intergenic region between H19 and L23mrp contains a boundary or an insulator element. We have identified a binding site for CTCF, a nuclear factor that mediates insulator activity in vertebrates, in the intergenic region. This site is conserved between human and mouse, associated with a major DNase I-hypersensitive site, and bound by CTCF in vivo. Functional assays using reporter constructs demonstrated that this element functions as an insulator in transfected cells. The findings suggest that this CTCF site contributes to the 3' boundary of this imprinted domain. Together with the findings on the differentially methylated CTCF sites 5' to H19, CTCF-dependent insulators may not only regulate but also delimit the imprinted domain. PMID- 12075008 TI - Muscle as a putative producer of acid alpha-glucosidase for glycogenosis type II gene therapy. AB - Glycogenosis type II (GSD II) is a lysosomal disorder affecting skeletal and cardiac muscle. In the infantile form of the disease, patients display cardiac impairment, which is fatal before 2 years of life. Patients with juvenile or adult forms can present diaphragm involvement leading to respiratory failure. The enzymatic defect in GSD II results from mutations in the acid alpha-glucosidase (GAA) gene, which encodes a 76 kDa protein involved in intralysosomal glycogen hydrolysis. We previously reported the use of an adenovirus vector expressing GAA (AdGAA) for the transduction of myoblasts and myotubes cultures from GSD II patients. Transduced cells secreted GAA in the medium, and GAA was internalized by receptor-mediated capture, allowing glycogen hydrolysis in untransduced cells. In this study, using a GSD II mouse model, we evaluated the feasibility of GSD II gene therapy using muscle as a secretary organ. Adenovirus vector encoding AdGAA was injected in the gastrocnemius of neonates. We detected a strong expression of GAA in the injected muscle, secretion into plasma, and uptake by peripheral skeletal muscle and the heart. Moreover, glycogen content was decreased in these tissues. Electron microscopy demonstrated the disappearance of destruction foci, normally present in untreated mice. We thus demonstrate for the first time that muscle can be considered as a safe and easily accessible organ for GSD II gene therapy. PMID- 12075009 TI - The Hook1 gene is non-functional in the abnormal spermatozoon head shape (azh) mutant mouse. AB - In mice carrying the autosomal recessive mutation 'abnormal spermatozoon head shape' (azh) all spermatozoa display a highly abnormal head morphology that differs drastically from the compact and hook-shaped head of the normal murine sperm. Moreover, the azh mutation causes tail abnormalities often resulting in coiled sperm tails or in the decapitation of the sperm head from the flagellum. We have isolated and characterized murine Hook1 cDNA and analyzed the corresponding genomic structure. Furthermore, the Hook1 gene was mapped to the same region on chromosome 4 to which the azh locus was previously linked. The Hook1 gene is predominantly expressed in haploid male germ cells, and immunohistochemical analysis revealed that Hook1 is responsible for the linkage of the microtubular manchette and the flagellum to cellular structures. Here, we report that the azh mutation is due to a deletion of exons 10 and 11 in the murine Hook1 gene leading to a non-functional protein. Our results indicate that loss of Hook1 function results in ectopic positioning of microtubular structures within the spermatid and causes the azh phenotype. Therefore, the human HOOK1 gene could serve as a candidate gene for male infertility due to teratozoospermia or decapitation defects. PMID- 12075010 TI - Evidence for translational regulation of the imprinted Snurf-Snrpn locus in mice. AB - In studies of genomic imprinting in the Prader-Willi/Angelman domain, an agouti coat color cassette was inserted into the downstream open reading frame (ORF) of the imprinted bicistronic Snurf-Snrpn locus in the mouse. The fusion gene was maternally silenced, as is Snurf-Snrpn, and produced a tan abdomen only when inherited paternally in otherwise-black mice. A screen for dominant epigenetic or genetic events was performed with ENU mutagenesis, using a strategy whereby variation in abdominal color was scored at weaning. One mouse with maternal origin of the fusion gene had a tan abdomen and had an imprinting defect resulting in loss of both maternal methylation and silencing of the fusion gene. One mouse with paternal origin of the fusion gene was completely yellow and was found to have an ATG-to-AAG mutation in the initiation codon of the upstream ORF encoding SNURF. Northern blotting, immunoblotting, and transfection studies indicated that the ATG-to-AAG mutation causes a 15-fold or more increase in translation of the downstream ORF in two fusion constructs, and it is likely that similar translational control affects the normal Snurf-Snrpn transcript as well. PMID- 12075011 TI - Nuclear genetic control of mitochondrial translation in skeletal muscle revealed in patients with mitochondrial myopathy. AB - Oxidative phosphorylation deficiencies can be caused by mutations in either the nuclear genome or the mitochondrial genome (mtDNA); however, most pathogenic mutations reported in adults occur in mtDNA. Such mutations often impair mitochondrial translation, and are associated with a characteristic muscle pathology consisting of a mosaic pattern of normal fibres interspersed with fibres that show mitochondrial proliferation (ragged-red fibres) and little or no complex IV (COX) activity. We investigated two adult patients with a severe mitochondrial myopathy in whom all muscle fibres showed mitochondrial proliferation with barely detectable COX activity - a pattern never before reported. Biochemical studies of the respiratory chain in muscle showed decreased activities of complexes I and IV (5% of control) and complex II+III (41% of control). Immunoblot analysis of nuclear and mitochondrial subunits of complexes I, III and IV showed a greater than 90% decrease in the steady-state level of these subunits in mature muscle, but no change in nuclear-encoded subunits of complexes II and V. A generalized mitochondrial translation defect was identified in pulse-label experiments in myotubes, but not in myoblasts cultured from both patients. This defect moved with the nucleus in patient cybrid cells. Myoblasts from one patient transplanted into the muscle bed of SCID mice differentiated into mature human muscle fibres that displayed a defect similar to that seen in the patient muscle. These results suggest a defect in a developmentally regulated nuclear factor important for mitochondrial translation in skeletal muscle. PMID- 12075013 TI - Exon discovery by genomic sequence alignment. AB - MOTIVATION: During evolution, functional regions in genomic sequences tend to be more highly conserved than randomly mutating 'junk DNA' so local sequence similarity often indicates biological functionality. This fact can be used to identify functional elements in large eukaryotic DNA sequences by cross-species sequence comparison. In recent years, several gene-prediction methods have been proposed that work by comparing anonymous genomic sequences, for example from human and mouse. The main advantage of these methods is that they are based on simple and generally applicable measures of (local) sequence similarity; unlike standard gene-finding approaches they do not depend on species-specific training data or on the presence of cognate genes in data bases. As all comparative sequence-analysis methods, the new comparative gene-finding approaches critically rely on the quality of the underlying sequence alignments. RESULTS: Herein, we describe a new implementation of the sequence-alignment program DIALIGN that has been developed for alignment of large genomic sequences. We compare our method to the alignment programs PipMaker, WABA and BLAST and we show that local similarities identified by these programs are highly correlated to protein-coding regions. In our test runs, PipMaker was the most sensitive method while DIALIGN was most specific. AVAILABILITY: The program is downloadable from the DIALIGN home page at http://bibiserv.techfak.uni-bielefeld.de/dialign/. PMID- 12075014 TI - A Bayesian network model for protein fold and remote homologue recognition. AB - MOTIVATION: The Bayesian network approach is a framework which combines graphical representation and probability theory, which includes, as a special case, hidden Markov models. Hidden Markov models trained on amino acid sequence or secondary structure data alone have been shown to have potential for addressing the problem of protein fold and superfamily classification. RESULTS: This paper describes a novel implementation of a Bayesian network which simultaneously learns amino acid sequence, secondary structure and residue accessibility for proteins of known three-dimensional structure. An awareness of the errors inherent in predicted secondary structure may be incorporated into the model by means of a confusion matrix. Training and validation data have been derived for a number of protein superfamilies from the Structural Classification of Proteins (SCOP) database. Cross validation results using posterior probability classification demonstrate that the Bayesian network performs better in classifying proteins of known structural superfamily than a hidden Markov model trained on amino acid sequences alone. PMID- 12075015 TI - Confidence measures for protein fold recognition. AB - MOTIVATION: We present an extensive evaluation of different methods and criteria to detect remote homologs of a given protein sequence. We investigate two associated problems: first, to develop a sensitive searching method to identify possible candidates and, second, to assign a confidence to the putative candidates in order to select the best one. For searching methods where the score distributions are known, p-values are used as confidence measure with great success. For the cases where such theoretical backing is absent, we propose empirical approximations to p-values for searching procedures. RESULTS: As a baseline, we review the performances of different methods for detecting remote protein folds (sequence alignment and threading, with and without sequence profiles, global and local). The analysis is performed on a large representative set of protein structures. For fold recognition, we find that methods using sequence profiles generally perform better than methods using plain sequences, and that threading methods perform better than sequence alignment methods. In order to assess the quality of the predictions made, we establish and compare several confidence measures, including raw scores, z-scores, raw score gaps, z score gaps, and different methods of p-value estimation. We work our way from the theoretically well backed local scores towards more explorative global and threading scores. The methods for assessing the statistical significance of predictions are compared using specificity--sensitivity plots. For local alignment techniques we find that p-value methods work best, albeit computationally cheaper methods such as those based on score gaps achieve similar performance. For global methods where no theory is available methods based on score gaps work best. By using the score gap functions as the measure of confidence we improve the more powerful fold recognition methods for which p values are unavailable. AVAILABILITY: The benchmark set is available upon request. PMID- 12075016 TI - Visualizing metabolic activity on a genome-wide scale. AB - MOTIVATION: To enhance the exploration of gene expression data in a metabolic context, one requires an application that allows the integration of this data and which represents this data in a (genome-wide) metabolic map. The layout of this metabolic map must be highly flexible to enable discoveries of biological phenomena. Moreover, it must allow the simultaneous representation of additional information about genes and enzymes. Since the layout and properties of existing maps did not fulfill our requirements, we developed a new way of representing gene expression data in metabolic charts. RESULTS: ViMAc generates user-specified (genome-wide) metabolic maps to explore gene expression data. To enhance the interpretation of these maps information such as sub-cellular localization is included. ViMAc can be used to analyse human or yeast expression data obtained with DNA microarrays or SAGE. We introduce our metabolic map method and demonstrate how it can be applied to explore DNA microarray data for yeast. AVAILABILITY: ViMAc is freely available for academic institutions on request from the authors. PMID- 12075017 TI - NETASA: neural network based prediction of solvent accessibility. AB - MOTIVATION: Prediction of the tertiary structure of a protein from its amino acid sequence is one of the most important problems in molecular biology. The successful prediction of solvent accessibility will be very helpful to achieve this goal. In the present work, we have implemented a server, NETASA for predicting solvent accessibility of amino acids using our newly optimized neural network algorithm. Several new features in the neural network architecture and training method have been introduced, and the network learns faster to provide accuracy values, which are comparable or better than other methods of ASA prediction. RESULTS: Prediction in two and three state classification systems with several thresholds are provided. Our prediction method achieved the accuracy level upto 90% for training and 88% for test data sets. Three state prediction results provide a maximum 65% accuracy for training and 63% for the test data. Applicability of neural networks for ASA prediction has been confirmed with a larger data set and wider range of state thresholds. Salient differences between a linear and exponential network for ASA prediction have been analysed. AVAILABILITY: Online predictions are freely available at: http://www.netasa.org. Linux ix86 binaries of the program written for this work may be obtained by email from the corresponding author. PMID- 12075018 TI - Modelling biological processes using workflow and Petri Net models. AB - MOTIVATION: Biological processes can be considered at many levels of detail, ranging from atomic mechanism to general processes such as cell division, cell adhesion or cell invasion. The experimental study of protein function and gene regulation typically provides information at many levels. The representation of hierarchical process knowledge in biology is therefore a major challenge for bioinformatics. To represent high-level processes in the context of their component functions, we have developed a graphical knowledge model for biological processes that supports methods for qualitative reasoning. RESULTS: We assessed eleven diverse models that were developed in the fields of software engineering, business, and biology, to evaluate their suitability for representing and simulating biological processes. Based on this assessment, we combined the best aspects of two models: Workflow/Petri Net and a biological concept model. The Workflow model can represent nesting and ordering of processes, the structural components that participate in the processes, and the roles that they play. It also maps to Petri Nets, which allow verification of formal properties and qualitative simulation. The biological concept model, TAMBIS, provides a framework for describing biological entities that can be mapped to the workflow model. We tested our model by representing malaria parasites invading host erythrocytes, and composed queries, in five general classes, to discover relationships among processes and structural components. We used reachability analysis to answer queries about the dynamic aspects of the model. AVAILABILITY: The model is available at http://smi.stanford.edu/projects/helix/pubs/process model/. PMID- 12075019 TI - Adaptive algorithm of automated annotation. AB - MOTIVATION: It is common knowledge that the avalanche of data arriving from the sequencing projects cannot be annotated either experimentally or manually by experts. The need for a reliable and convenient tool for automated sequence annotation is broadly recognized. RESULTS: Here, we describe the Adaptive Algorithm of Automated Annotation (A(4)) based on a statistical approach to this problem. The mathematical model relates a set of homologous sequences and descriptions of their functional properties, and calculates the probabilities of transferring a sequence description onto its homologue. The proposed model is adaptive, its parameters (distribution characteristics, transference probabilities, thresholds, etc.) are dynamic, i.e. are generated individually for the sequences and various functional properties (words of the description). The proposed technique significantly outperforms the widely used test for frequency threshold, which is a special case of our model realized for the simplest set of parameters. The prediction technique has been realized as a computer program and tested on a random sequence sampling from SWISS-PROT. AVAILABILITY: The automated annotation program based on the proposed algorithm is available through the Web browser at http://www.genebee.msu.su/services/annot/basic.html. PMID- 12075020 TI - A novel approach to local reliability of sequence alignments. AB - MOTIVATION: The pairwise alignment of biological sequences obtained from an algorithm will in general contain both correct and incorrect parts. Hence, to allow for a valid interpretation of the alignment, the local trustworthiness of the alignment has to be quantified. RESULTS: We present a novel approach that attributes a reliability index to every pair of residues, including gapped regions, in the optimal alignment of two protein sequences. The method is based on a fuzzy recast of the dynamic programming algorithm for sequence alignment in terms of mean field annealing. An extensive evaluation with structural reference alignments not only shows that the probability for a pair of residues to be correctly aligned grows consistently with increasing reliability index, but moreover demonstrates that the value of the reliability index can directly be translated into an estimate of the probability for a correct alignment. PMID- 12075021 TI - The chemical organization of signaling interactions. AB - MOTIVATION: Cellular chemical signaling pathways form complex networks that are beginning to be studied at the level of chemical kinetics and databases of reactions. Chemical reaction details are traditionally represented as lists of reactions and rates. This does not map readily to the block diagram representation familiar to biologists, and obscures the functional organization of signaling networks. This study examines motifs in signaling chemistry and reports common features that may help to formalize such a mapping between pathway block diagrams and the chemistry. The same motifs may facilitate data representation and provide functional abstraction of the chemistry. RESULTS: I classified 74 interactions between 25 signaling pathways in terms of shared chemical motifs. All interactions in this dataset consist of a few communicating molecules from one set of pathways, and a replicating set of reactions and molecules from another. Each unique combination of interacting pathways duplicates the chemical reaction scheme of this replicating set, but involves different rate constants. Signaling pathways can therefore be described in an object-oriented manner as sets of core reactions with well-defined interfaces between pathways. This generalization lends itself to designing simulators and databases for signaling networks. AVAILABILITY: Software and example models are freely available from http://www.ncbs.res.in/~bhalla/examples/EGFR_example.html. PMID- 12075022 TI - Hybrid alignment: high-performance with universal statistics. AB - The score statistics of a recently introduced 'hybrid alignment' algorithm is studied in detail numerically. An extensive survey across the 2216 models of protein domains contained in the Pfam v5.4 database (Bateman et al., Nucleic Acids Res., 28, 263-266, 2000) verifies the theoretical predictions: For the position-specific scoring functions used in the Pfam models, the score statistics of hybrid alignment obey the Gumbel distribution, with the key Gumbel parameter lambda taking on the asymptotic value 1 universally for all models. Thus, the use of hybrid alignment eliminates the time-consuming computer simulations normally needed to assign p-values to alignment scores, freeing the users to experiment with different scoring parameters and functions. The performance of the hybrid algorithm in detecting sequence homology is also studied. For protein sequences from the SCOP database (Murzin et al., J. Mol. Biol., 247, 536-540, 1995) using uniform scoring functions, the performance is found to be comparable to the best of the existing methods. Preliminary results using the PfamA database suggest that the hybrid algorithm achieves similar performance as existing methods for position-specific scoring systems as well. Hybrid alignment is thereby established as a high performance alignment algorithm with well-characterized, universal statistics. PMID- 12075023 TI - SST: an algorithm for finding near-exact sequence matches in time proportional to the logarithm of the database size. AB - MOTIVATION: Searches for near exact sequence matches are performed frequently in large-scale sequencing projects and in comparative genomics. The time and cost of performing these large-scale sequence-similarity searches is prohibitive using even the fastest of the extant algorithms. Faster algorithms are desired. RESULTS: We have developed an algorithm, called SST (Sequence Search Tree), that searches a database of DNA sequences for near-exact matches, in time proportional to the logarithm of the database size n. In SST, we partition each sequence into fragments of fixed length called 'windows' using multiple offsets. Each window is mapped into a vector of dimension 4(k) which contains the frequency of occurrence of its component k-tuples, with k a parameter typically in the range 4-6. Then we create a tree-structured index of the windows in vector space, with tree structured vector quantization (TSVQ). We identify the nearest neighbors of a query sequence by partitioning the query into windows and searching the tree structured index for nearest-neighbor windows in the database. When the tree is balanced this yields an O(logn) complexity for the search. This complexity was observed in our computations. SST is most effective for applications in which the target sequences show a high degree of similarity to the query sequence, such as assembling shotgun sequences or matching ESTs to genomic sequence. The algorithm is also an effective filtration method. Specifically, it can be used as a preprocessing step for other search methods to reduce the complexity of searching one large database against another. For the problem of identifying overlapping fragments in the assembly of 120 000 fragments from a 1.5 megabase genomic sequence, SST is 15 times faster than BLAST when we consider both building and searching the tree. For searching alone (i.e. after building the tree index), SST 27 times faster than BLAST. AVAILABILITY: Request from the authors. PMID- 12075024 TI - Global analysis of tandem aromatic octapeptide repeats: the significance of the aromatic-glycine motif. AB - MOTIVATION: Tandem peptide repeats play a key role in self-assembly and aggregation processes. A notable example is the occurrence of tandem peptide repeats in prionic proteins and their role in the aggregation process that leads to the formation of the prion. One of the structural characteristics that is evident from the comparison of mammalian and yeast prion proteins is the presence of aromatic residues in their tandem repeats. These residues are accompanied by glycine residues before and/or after the aromatic amino acid. Such aromatic glycine conjugates are also present in the tandem repeats of the large family of the bacterial ice nucleation proteins. To study the significance of such aromatic glycine occurrences, a global analysis of all the aromatic octapeptide repeats in the Swiss-Prot and TrEMBL databases was conducted. The search pattern was formulated to compare the number of conjugates of each of the 20 natural amino acids before or after the different aromatic residues. RESULTS: The presence of aromatic-glycine conjugates appears to be significantly higher than aromatic conjugates to any other amino acid. Furthermore, all the six various combination of glycine occurrences before or after the three aromatic residues are present. No such pattern was observed for any other amino acid. The significance of the findings is being discussed in the context of the physicochemical properties of aromatic-glycine conjugates and its possible role in the facilitation of aggregates formation. PMID- 12075025 TI - AiO, combining DNA/protein programs and oligo-management. AB - AiO (All in One) is a program for Windows, that combines typical DNA/protein features such as plasmid map drawing, finding of ORFs, translate, backtranslate and high quality printing with a number of databases. These databases allow the management of oligonucleotides, oligonucleotide-manufacturers, restriction enzymes, structural DNA and program users in a multi-user/multi-group environment. AVAILABILITY: An AiO specific website, with the possibility to download is at: http://134.99.88.55/aio/ SUPPLEMENTARY INFORMATION: Examples with screen shots- http://134.99.88.55/aio/ : Manual (in PDF format) http://134.99.88.55/aio/manual.pdf PMID- 12075026 TI - Synonymous-non-synonymous mutation rates between sequences containing ambiguous nucleotides (Syn-SCAN). AB - SUMMARY: Direct PCR sequencing on genetic material containing allelic mixtures results in sequences containing ambiguous nucleotides. Because codons exhibiting allelic mixtures present evidence of evolutionary pressure, it is important to include this information in the assessment of codon synonymy. We developed a program, 'Synonymous-Nonsynonymous Mutation Rates between Sequences Containing Ambiguous Nucleotides' (Syn-SCAN), that calculates synonymous and non-synonymous substitution rates using a model that includes allelic mixtures. AVAILABILITY: Syn-SCAN is implemented on the web and can be downloaded from http://hivdb.stanford.edu. PMID- 12075027 TI - Arrayplot for visualization and normalization of cDNA microarray data. AB - Arrayplot is an application which allows filtering, visualization and normalization of raw cDNA microarray data. AVAILABILITY: MS-Windows binaries are freely available for non-profit use at http://www.biologie.ens.fr/yeast publi.html. SUPPLEMENTARY INFORMATION: Scripts for online creation of Arrayplot files are available at http://transcriptome.ens.fr/lgm_bioinfo/ PMID- 12075028 TI - MOUSE (Mitochondrial and Other Useful SEquences) a compilation of population genetic markers. AB - Mitochondrial and Other Useful SEquences (MOUSE) is an integrated and comprehensive compilation of mtDNA from hypervariable regions I and II and of the low recombining nuclear loci Xq13.3 from about 11 200 humans and great apes, whose geographic and if applicable, linguistic classification is stored with their aligned sequences and publication details. The goal is to provide population geneticists and genetic epidemiologists with a comprehensive and user friendly repository of sequences and population information that is usually dispersed in a variety of other sources. AVAILABILITY: http://www.gen epi.de/mouse. SUPPLEMENTARY INFORMATION: Documentation and detailed information on population subgroups is available on the homepage: http://www.gen-epi.de/mouse PMID- 12075029 TI - BAOBAB: a Java editor for large phylogenetic trees. AB - SUMMARY: BAOBAB is a Java user interface dedicated to viewing and editing large phylogenetic trees. Original features include: (i) a colour-mediated overview of magnified subtrees; (ii) copy/cut/paste of (sub)trees within or between windows; (iii) compressing/ uncompressing subtrees; and (iv) managing sequence files together with tree files. AVAILABILITY: http://www.univ-montp2.fr/~genetix/. PMID- 12075030 TI - DMLE+: Bayesian linkage disequilibrium gene mapping. AB - SUMMARY: The program DMLE+ allows Bayesian inference of the location of a gene carrying a mutation influencing a discrete trait (such as a disease) and/or other parameters of interest (such as mutation age) based on the observed linkage disequilibrium at multiple genetic markers. DMLE+ uses either individual marker genotypes, or haplotypes, integrates over uncertain population allele frequencies, and can incorporate prior information about gene location from an annotated human genome sequence. AVAILABILITY: DMLE+ is available in both Windows GUI and portable UNIX command line versions at http://dmle.org. PMID- 12075031 TI - The role of atypical antipsychotics in the treatment of delirium. AB - Delirium exemplifies the interface between medicine and psychiatry. It is generally characterized by acute disturbances of consciousness, cognition, and perception that are precipitated by an underlying medical condition. The gold standard of psychiatric treatment is to treat the underlying medical cause and use high-potency antipsychotics to treat the clinical manifestations of delirium. In the early 1990s, a new generation of novel antipsychotics was developed. Their mechanism of action, preferential serotonergic (5HT(2a)) blockade, results in a markedly lower rate of extrapyramidal side effects, an advantage over the typical, older antipsychotic medications. These agents have been shown to be effective and well tolerated in common psychotic disorders (e.g., schizophrenia or bipolar disorder), but few studies have evaluated them in the treatment of delirium. This paper reviews the pertinent literature and summarizes tentative guidelines for novel antipsychotic use in delirium. PMID- 12075032 TI - An open trial of olanzapine for the treatment of delirium in hospitalized cancer patients. AB - We conducted an open, prospective trial of olanzapine for the treatment of delirium in a sample of 79 hospitalized cancer patients. Patients all met DSM-IV criteria for a diagnosis of delirium and were rated systematically with the Memorial Delirium Assessment Scale (MDAS) as a measure of delirium severity, phenomenology, and resolution, over the course of a 7-day treatment period. Sociodemographic and medical variables and measures of physical performance status and drug-related side effects were collected. Fifty-seven patients (76%) had complete resolution of their delirium on olanzapine therapy. No patients experienced extrapyramidal side effects; however, 30% experienced sedation (usually not severe enough to interrupt treatment). Several factors were found to be significantly associated with poorer response to olanzapine treatment for delirium, including age >70 years, history of dementia, central nervous system spread of cancer and hypoxia as delirium etiologies, "hypoactive" delirium, and delirium of "severe" intensity (i.e., MDAS >23). A logistic-regression model suggests that age >70 years is the most powerful predictor of poorer response to olanzapine treatment for delirium (odds ratio, 171.5). Olanzapine appears to be a clinically efficacious and safe drug for the treatment of the symptoms of delirium in the hospitalized medically ill. PMID- 12075033 TI - The delirium experience: delirium recall and delirium-related distress in hospitalized patients with cancer, their spouses/caregivers, and their nurses. AB - We conducted a systematic examination of the experience of delirium in a sample of 154 hospitalized patients with cancer. Patients all met DSM-IV criteria for delirium and were rated with the Memorial Delirium Assessment Scale as a measure of delirium severity, phenomenology, and resolution. Of the 154 patients assessed, 101 had complete resolution of their delirium and were administered the Delirium Experience Questionnaire (DEQ-a face-valid measure that assesses delirium recall and distress related to the delirium episode). Spouse/caregivers and primary nurses were also administered the DEQ to assess distress related to caring for a delirious patient. Fifty-four (53.5%) patients recalled their delirium experience. Logistic-regression analysis demonstrated that short-term memory impairment (odds ratio [OR] = 38.4), delirium severity (OR = 11.3), and the presence of perceptual disturbances (OR = 6.9) were significant predictors of delirium recall. Mean delirium-related distress levels (on a 0-4 numerical rating scale of the DEQ) were 3.2 for patients who recalled delirium, 3.75 for spouses/caregivers, and 3.09 for nurses. Logistic-regression analysis demonstrated that the presence of delusions (OR = 7.9) was the most significant predictor of patient distress. Patients with "hypoactive" delirium were just as distressed as patients with "hyperactive" delirium. Karnofsky Performance Status (OR = 9.1) was the most significant predictor of spouse/caregiver distress. Delirium severity (OR =5.2) and the presence of perceptual disturbances (OR =3.6) were the most significant predictors of nurse distress. In conclusion, a majority of patients with delirium recall their delirium as highly distressing. Delirium is also a highly distressing experience for spouses/caregivers and nurses who are caring for delirious patients. Prompt recognition and treatment of delirium is critically important to reduce suffering and distress. PMID- 12075034 TI - Posttraumatic stress disorder and self-reported physical health status among U.S. Military personnel serving during the Gulf War period: a population-based study. AB - The objective of this study was to investigate the relation between posttraumatic stress disorder (PTSD) and perceived physical health. Participants included 3,682 Gulf War veterans and control subjects of the same era who completed a telephone survey about their health status. PTSD was assessed using the PTSD Checklist Military Version. Veterans screening positive for PTSD reported significantly more physical health symptoms and medical conditions than did veterans without PTSD. They were also more likely to rate their health status as fair or poor and to report lower levels of health-related quality of life. The results of this study are consistent with studies of other combat veterans and provide further support for an association between PTSD and adverse physical health outcomes. Stressful or traumatic life events, such as those encountered during a rapid military deployment and conflict, are associated with a variety of adverse health effects. These health effects may manifest themselves in both psychological and physical outcomes. Health care providers must be attentive to recognize and evaluate both of these dimensions. PMID- 12075035 TI - Association of attachment style to lifetime medically unexplained symptoms in patients with hepatitis C. AB - In this study, we sought to establish whether there was an association between adult attachment style and number of medically unexplained physical symptoms in patients with hepatitis C. Thirty-two patients with hepatitis C were assessed with regard to attachment style classification, number of lifetime medically unexplained symptoms, lifetime psychiatric diagnoses, medical comorbidity, disease severity, use of interferon, and demographic characteristics. Analysis of covariance was used to compare the four attachment groups on number of lifetime medically unexplained symptoms, and Pearson correlations were used to assess the association of continuous ratings of attachment style with lifetime medically unexplained symptoms. Number of lifetime medically unexplained symptoms varied significantly as a function of attachment style group, with patients with fearful attachment reporting significantly more medically unexplained symptoms than patients with secure attachment (P < 0.01). Number of lifetime medically unexplained symptoms was positively correlated with continuous ratings of fearful attachment (r = 0.53, P < 0.01) and preoccupied attachment (r = 0.46, P < 0.01). Implications for treatment are discussed. PMID- 12075036 TI - Spirituality, religion, and depression in the terminally ill. AB - OBJECTIVE: This study examined the impact of spirituality and religiosity on depressive symptom severity in a sample of terminally ill patients with cancer and AIDS. METHODS: One hundred sixty-two patients were recruited from palliative care facilities (hospitals and specialized nursing facilities), all of whom had a life expectancy <6 months. The primary variables used in this study were the FACIT Spiritual Well-Being Scale, a religiosity index similar to those used in previous research, the Hamilton Depression Rating Scale (HDRS), the Karnofsky Performance Rating Scale, the Memorial Symptom Assessment Scale, and the Duke-UNC Functional Social Support Questionnaire. RESULTS: A strong negative association was observed between the FACIT Spiritual Well-Being scale and the HDRS, but no such relationship was found for religiosity, because more religious individuals had somewhat higher scores on the HDRS. Similar patterns were observed for the FACIT subscales, finding a strong negative association between the meaning and peace subscale (which corresponds to the more existential aspects of spirituality) and HDRS scores, whereas a positive, albeit nonsignificant, association was observed for the faith subscale (which corresponds more closely to religiosity). CONCLUSIONS: These results suggest that the beneficial aspects of religion may be primarily those that relate to spiritual well-being rather than to religious practices per se. Implications for clinical interventions and palliative-care practice are discussed. PMID- 12075037 TI - Consultation-liaison psychiatrists' use of antidepressants in the physically ill. AB - In a practice-based, prospective study of 917 inpatients referred to a consultation-liaison psychiatry service and diagnosed as depressed, 41% were prescribed an antidepressant: 40% tricyclics, 35% selective serotonin reuptake inhibitors (SSRIs), 15% monoamine oxidase inhibitors (MAOIs)/reversible inhibitors of monoamine (RIMAs) (mainly moclobemide), and 11% tetracyclics (mianserin). Factors associated with choice of antidepressant type included age, referral for pain, length and seriousness of physical illness, type of physical illness, and concurrent antipsychotics (P < 0.01). Tetracyclics and MAOI/RIMAs were used significantly more often than tricyclics in the more severely physically ill and the elderly. The percentage of patients prescribed an antidepressant increased significantly over time, which is accounted for by the greater use of SSRIs across all age groups and degrees of seriousness of illness. There is a paucity of randomized controlled trials on which to base practice guidelines. Practice-based research such as this helps inform those guidelines. PMID- 12075038 TI - Adverse reactions to antidepressants in consultation-liaison psychiatry inpatients. AB - In a practice-based, prospective study of 1,551 inpatients referred to a consultation-liaison psychiatry service and prescribed an antidepressant, an adverse drug reaction judged sufficient to warrant discontinuation of the drug was noted in 158 (10.2%). The factors associated were older age (P < 0.05); ICD-9 genitourinary disorder (mainly renal failure) (P < 0.01); DSM-IV "delirium, dementia, etc." (P < 0.05); length of stay (P < 0.001); number of visits (P < 0.001); and time spent on case (P < 0.05). Neither psychological functioning (GAF) nor physical functioning (Karnofsky ratings) were associated with a reaction, nor was multiple psychotropic drug prescription. Tricyclics were more likely than selective serotonin reuptake inhibitors/norepinephrine reuptake inhibitors to be associated with a reaction (P < 0.05). Consultation-liaison psychiatrists need to be well informed about pharmacodynamics and drug interactions in patients with physical/psychiatric comorbidity. PMID- 12075039 TI - Acute psychosis in a patient with vitamin B(12) deficiency and coincident cervical stenosis. PMID- 12075040 TI - Neuropsychiatric complications associated with interferon-alpha-2b treatment of malignant melanoma. PMID- 12075041 TI - A case of SIADH and hyponatremia associated with citalopram. PMID- 12075042 TI - Stiff-man syndrome: a case report and review of the literature. PMID- 12075043 TI - Med-psych drug-drug interactions update. PMID- 12075044 TI - Linezolid: MAOI activity and potential drug interactions. PMID- 12075045 TI - Clozapine-associated cardiomyopathy. PMID- 12075046 TI - The origin of the concept of somatization. PMID- 12075047 TI - Pretreatment ECGs and the prescription of amitriptyline in an internal medicine clinic. PMID- 12075048 TI - Psychogenic excoriation and cancer. PMID- 12075053 TI - Myocardial ischemia and the pains of the heart. PMID- 12075054 TI - The use of molecular profiling to predict survival after chemotherapy for diffuse large-B-cell lymphoma. AB - BACKGROUND: The survival of patients with diffuse large-B-cell lymphoma after chemotherapy is influenced by molecular features of the tumors. We used the gene expression profiles of these lymphomas to develop a molecular predictor of survival. METHODS: Biopsy samples of diffuse large-B-cell lymphoma from 240 patients were examined for gene expression with the use of DNA microarrays and analyzed for genomic abnormalities. Subgroups with distinctive gene-expression profiles were defined on the basis of hierarchical clustering. A molecular predictor of risk was constructed with the use of genes with expression patterns that were associated with survival in a preliminary group of 160 patients and was then tested in a validation group of 80 patients. The accuracy of this predictor was compared with that of the international prognostic index. RESULTS: Three gene expression subgroups--germinal-center B-cell-like, activated B-cell-like, and type 3 diffuse large-B-cell lymphoma--were identified. Two common oncogenic events in diffuse large-B-cell lymphoma, bcl-2 translocation and c-rel amplification, were detected only in the germinal-center B-cell-like subgroup. Patients in this subgroup had the highest five-year survival rate. To identify other molecular determinants of outcome, we searched for individual genes with expression patterns that correlated with survival in the preliminary group of patients. Most of these genes fell within four gene-expression signatures characteristic of germinal-center B cells, proliferating cells, reactive stromal and immune cells in the lymph node, or major-histocompatibility-complex class II complex. We used 17 genes to construct a predictor of overall survival after chemotherapy. This gene-based predictor and the international prognostic index were independent prognostic indicators. CONCLUSIONS: DNA microarrays can be used to formulate a molecular predictor of survival after chemotherapy for diffuse large-B-cell lymphoma. PMID- 12075055 TI - Abnormal subendocardial perfusion in cardiac syndrome X detected by cardiovascular magnetic resonance imaging. AB - BACKGROUND: In cardiac syndrome X (a syndrome characterized by typical angina, abnormal exercise-test results, and normal coronary arteries), conventional investigations have not found that chest pain is due to myocardial ischemia. Magnetic resonance techniques have higher resolution and therefore may be more sensitive. METHODS: We performed myocardial-perfusion cardiovascular magnetic resonance imaging in 20 patients with syndrome X and 10 matched controls, both at rest and during an infusion of adenosine. Quantitative perfusion analysis was performed by using the normalized upslope of myocardial signal enhancement to derive the myocardial perfusion index and the myocardial-perfusion reserve index (defined as the ratio of the myocardial perfusion index during stress to the index at rest). RESULTS: In the controls, the myocardial perfusion index increased in both myocardial layers with adenosine (in the subendocardium, from a mean [+/-SD] of 0.12+/-0.03 to 0.16+/-0.03 [P=0.02]; in the subepicardium, from 0.11+/-0.02 to 0.17+/-0.05 [P=0.002]); in patients with syndrome X, the myocardial perfusion index did not change significantly in the subendocardium (0.13+/-0.02 vs. 0.14+/-0.03, P=0.11; P=0.09 as compared with controls) but increased in the subepicardium (from 0.11+/-0.02 to 0.20+/-0.04, P<0.001; P=0.11 for the comparison with controls). Adenosine provoked chest pain in 95 percent of patients with syndrome X and 40 percent of controls (P<0.001). CONCLUSIONS: In patients with syndrome X, cardiovascular magnetic resonance imaging demonstrates subendocardial hypoperfusion during the intravenous administration of adenosine, which is associated with intense chest pain. These data support the notion that the chest pain may have an ischemic cause. PMID- 12075056 TI - Endothelial function and oxidative stress in renovascular hypertension. AB - BACKGROUND: It has been reported that renovascular hypertension activates the renin-angiotensin system, leading to an increase in oxidative stress. We sought to determine whether renal-artery angioplasty improves endothelial dysfunction in patients with renovascular hypertension through a reduction in oxidative stress. METHODS: We evaluated the response of forearm blood flow to acetylcholine, an endothelium-dependent vasodilator, and isosorbide dinitrate, an endothelium independent vasodilator, before and after renal-artery angioplasty in 15 subjects with renovascular hypertension and 15 controls without hypertension who were matched for age and sex. Forearm blood flow was measured with the use of a mercury-filled Silastic strain-gauge plethysmograph. RESULTS: The forearm blood flow in response to acetylcholine was less in subjects with renovascular hypertension than in controls, although the forearm blood flow in response to isosorbide dinitrate was similar in the two groups. Angioplasty decreased systolic and diastolic blood pressures, forearm vascular resistance, and urinary excretion of 8-hydroxy-2'-deoxyguanosine and serum malondialdehyde-modified low density lipoprotein (LDL), indexes of oxidative stress. After angioplasty, the mean (+/-SD) forearm blood flow in response to acetylcholine was increased in the patients with renovascular hypertension (19.3+/-6.8 vs. 29.6+/-7.1 ml per minute per 100 ml, P=0.002). The increase in the maximal forearm blood flow in response to acetylcholine correlated significantly with the decrease in urinary excretion of 8-hydroxy-2'-deoxyguanosine (r=-0.51, P=0.004) and serum malondialdehyde modified LDL (r=-0.39, P=0.02). Coinfusion of ascorbic acid (vitamin C) augmented the response of forearm blood flow to acetylcholine before angioplasty (P<0.001) but not after angioplasty. CONCLUSIONS: These findings suggest that excessive oxidative stress is involved, at least in part, in impaired endothelium-dependent vasodilatation in patients with renovascular hypertension. PMID- 12075057 TI - Fumagillin treatment of intestinal microsporidiosis. AB - BACKGROUND: Intestinal microsporidiosis due to Enterocytozoon bieneusi is a cause of chronic diarrhea, malabsorption, and wasting in immunocompromised patients. Currently, there is no effective treatment. METHODS: We conducted a randomized, double-blind, placebo-controlled trial of fumagillin (60 mg per day orally for two weeks) in patients with chronic E. bieneusi infection. Efficacy was assessed primarily by the clearance of microsporidia, as evidenced by analysis of stool specimens. Patients in whom microsporidia were not cleared received treatment for two weeks with open-label fumagillin. After clearance of the parasite, follow-up stool examinations were performed monthly to detect relapses. RESULTS: Twelve patients were enrolled in this study, 10 with the acquired immunodeficiency syndrome and 2 who had received organ transplants. Clearance of microsporidia occurred in all six of the patients in the fumagillin group, as compared with none of the six in the placebo group (P=0.002). Treatment with fumagillin was also associated with increases in absorption of D-xylose (P=0.003) and in Karnofsky performance scores (P<0.001) and with decreases in loperamide use (P=0.01) and total stool weight (P=0.04). There were serious adverse events (neutropenia and thrombocytopenia) in three patients in the fumagillin group; one patient in the placebo group had severe diarrhea. All six controls subsequently had clearance of microsporidia after open-label treatment with fumagillin. Relapses of the infection were identified in two patients during follow-up (median follow-up, 10 months). CONCLUSIONS: Fumagillin is an effective treatment for chronic E. bieneusi infection in immunocompromised patients. PMID- 12075058 TI - Images in clinical medicine. Nodular sarcoidosis. PMID- 12075059 TI - Clinical practice. Pleural effusion. PMID- 12075060 TI - Heat stroke. PMID- 12075061 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 19-2002. A 13-year-old girl with a mass in the left parotid gland and regional lymph nodes. PMID- 12075062 TI - Molecular characteristics of diffuse large-B-cell lymphoma. PMID- 12075063 TI - Hypertension, angiotensin II, and oxidative stress. PMID- 12075065 TI - Homocysteine and dementia. PMID- 12075064 TI - Ethical incentives--not payment--for organ donation. PMID- 12075066 TI - Leptin-replacement therapy in lipodystrophy. PMID- 12075067 TI - Direct-to-consumer marketing. PMID- 12075068 TI - Malposition of a pacemaker lead. PMID- 12075069 TI - Debt repayment for trainees. PMID- 12075070 TI - Adult T-cell leukemia-lymphoma during pregnancy. PMID- 12075072 TI - Dissecting virus entry via endocytosis. AB - Numerous virus families utilize endocytosis to infect host cells, mediating virus internalization as well as trafficking to the site of replication. Recent research has demonstrated that viruses employ the full endocytic capabilities of the cell. The endocytic pathways utilized include clathrin-mediated endocytosis, caveolae, macropinocytosis and novel non-clathrin, non-caveolae pathways. The tools to study endocytosis and, consequently, virus entry are becoming more effective and specific as the amount of information on endocytic component structure and function increases. The use of inhibitory drugs, although still quite common, often leads to non-specific disruptions in the cell. Molecular inhibitors in the form of dominant-negative proteins have surpassed the use of chemical inhibitors in terms of specificity to individual pathways. Dominant negative molecules are derived from both structural proteins of endocytosis, such as dynamin and caveolin, and regulatory proteins, primarily small GTPases and kinases. This review focuses on the experimental approaches taken to examine virus entry and provides both classic examples and recent research on a variety of virus families. PMID- 12075073 TI - A time to kill: viral manipulation of the cell death program. AB - Many viruses have as part of their arsenal the ability to modulate the apoptotic pathways of the host. It is counter-intuitive that such simple organisms would be efficient at regulating this the most crucial pathway within the host, given the relative complexity of the host cells. Yet, viruses have the potential to initiate or stay the onset of programmed cell death through the manipulation of a variety of key apoptotic proteins. It is the intention of this review to provide an overview of viral gene products that are able to promote or inhibit apoptotic death of the host cell and to discuss their mechanisms of action. It is not until recently that the depth at which viruses exploit the apoptotic pathways of their host has been seen. This understanding may provide a great opportunity for future therapeutic ventures. PMID- 12075074 TI - Lack of involvement of haemocytes in the establishment and spread of infection in Spodoptera frugiperda larvae infected with the baculovirus Autographa californica M nucleopolyhedrovirus by intrahaemocoelic injection. AB - It is thought that insect haemocytes, or blood cells, play an important role in baculovirus pathogenesis by amplifying and helping to spread the infection within the insect. Here, infection is described of the lepidopteran noctuid species Spodoptera frugiperda with the baculovirus Autographa californica M nucleopolyhedrovirus (AcMNPV). Late instar S. frugiperda larvae were infected by intrahaemocoelic injection using a recombinant of AcMNPV expressing the enhanced green fluorescent protein gene to visualize infected cells. Approximately 1000 fold higher doses of injected virus were required to initiate infection in S. frugiperda larvae than in another permissive noctuid species, Trichoplusia ni. Infected S. frugiperda larvae survived twice as long as T. ni larvae and exhibited a slower build-up of virus in the haemolymph. In S. frugiperda, infection of fat body and epithelium was observed prior to significant infection of haemocytes, even though the virus was delivered by intrahaemocoelic injection. Expression of eGFP was first detected 12-18 h post-injection within the fat body and, by 24 h, infection had spread to the tracheal and body wall epithelium. In contrast, only 5% of haemocytes were infected at 24 h and the proportion of infected haemocytes increased slowly to only around 50% at 5 days post-infection, when most larval death occurred. Thus, in S. frugiperda, haemocytes do not appear to have a primary role in AcMNPV pathogenesis. This relative lack of infection of haemocytes may in part explain why S. frugiperda larvae are more resistant to AcMNPV infection than T. ni larvae. PMID- 12075075 TI - Homologous recombination between the inverted terminal repeats of defective transposon TCp3.2 causes an inversion in the genome of Cydia pomonella granulovirus. AB - In this study, a new mutant of the Cydia pomonella granulovirus (CpGV), which shows spontaneous inversion of a transposable element during in vivo replication, is described. CpGV-MCp4 is a natural mutant of CpGV-M, containing the transposable element TCp3.2, which originated from the genome of the host C. pomonella. During in vivo cloning studies of CpGV-MCp4, a mutant called CpGV MCp4inv was isolated. CpGV-MCp4inv shows heterogeneity in the genome area of transposon insertion. Restriction mapping, PCR analysis and subsequent sequence analysis gave strong evidence that an inversion of TCp3.2 is caused by homologous recombination between the long inverted terminal repeats (ITRs) of the transposon. This finding demonstrated that extensive homologous repeat regions such as the ITRs of transposons cause inversions by homologous recombination during in vivo replication. The observed in vivo inversion between the ITRs can be considered as a model for the contribution of repeated sequences in the genome rearrangement of baculoviruses and a source for genetic heterogeneity among different baculoviruses and baculovirus genotypes. PMID- 12075076 TI - Primary herpes simplex virus type 1 infection of the eye triggers similar immune responses in the cornea and the skin of the eyelids. AB - Herpetic stromal keratitis (HSK) and blepharoconjunctivitis in humans are thought partly to result from immunopathological responses to herpes simplex virus type 1 (HSV-1). The corneas of NIH mice were inoculated with HSV-1 (strain McKrae) and mice were examined for signs of disease and infection on days 1, 4, 7, 10, 14 and 21. The eyes and eyelids of infected and control mice were processed for immunohistochemistry and double stained for viral antigens and one of the following cell surface markers (Gr-1, F4/80, CD4, CD8, CD45R or MHC class II) or one of the following cytokines (IL-2, IL-4, IL-6, IL-10, IL-12 or IFN-gamma). All infected mice developed signs of HSK by day 4 and blepharitis by day 7 and these both persisted until day 21, when signs of resolution where apparent. Virus was detected during the first week of infection and became undetectable by day 10. Large numbers of Gr-1(+) cells (neutrophils) infiltrated infected corneas and eyelids in areas of viral antigen and CD4(+) T cells increased significantly in number after virus clearance. In both sites, the predominant cytokines were IL-6, IL-10, IL-12 and IFN-gamma, with few IL-2(+) and IL-4(+) cells. These observations suggest that the immune responses in the cornea are similar to those in the eyelids but, overall, the responses are not clearly characterized as either Th1 or Th2. In both sites, the neutrophil is the predominant infiltrating cell type and is a likely source of the cytokines observed and a major effector of the disease process. PMID- 12075077 TI - Herpes simplex virus type 1 infection prevents detachment of nerve growth factor differentiated PC12 cells in culture. AB - In tissue culture, rat pheochromocytoma (PC12) cells differentiated with nerve growth factor (NGF) cease division, extend neuritic processes and acquire many properties characteristic of neuronal cells. In previous work, we have shown that NGF-differentiated PC12 cells can survive infection with herpes simplex virus type 1 (HSV-1) and maintain the viral genome in a quiescent but reactivatable state. In this study, we report that uninfected NGF-differentiated PC12 cells uniformly and predictably detach from the culture flask substratum after approximately 7 weeks. Although uninfected cells were uniformly lost from the culture by 7 weeks, surprisingly HSV-1-infected cells survived beyond 10 weeks, the time limit of the study. The detachment of uninfected cells was not the result of cell death or apoptosis, as determined by viability assays performed on cells after detachment. Expression of the HSV-1 latency associated transcript (LAT) gene and virus replication was not necessary for the virus to suppress the 'detachment' phenomenon, since NGF-differentiated PC12 cells infected with either wild-type, DNA polymerase mutant or LAT null mutant virus survived in culture for similar lengths of time. Viral gene expression does appear to be necessary for the suppression, however, since cells infected with UV-inactivated virus were lost from culture with kinetics similar to those of uninfected cells. These findings indicate that de novo viral gene synthesis mediates changes to the host NGF-differentiated PC12 cells, which results in prevention of detachment. PMID- 12075078 TI - Activity and intracellular localization of the human cytomegalovirus protein pp71. AB - The human cytomegalovirus (HCMV) tegument phosphoprotein pp71 activates viral immediate early (IE) transcription and thus has a role in initiating lytic infection. Protein pp71 stimulates expression from a range of promoters in a sequence-independent manner, and in this respect behaves similarly to the herpes simplex virus type 1 (HSV-1) IE protein ICP0. The intracellular localization of pp71 was investigated after its expression from transfected plasmids or from HSV 1 mutants constructed to produce pp71 transiently. The protein colocalized with the cell promyelocytic leukaemia (PML) protein at nuclear domain 10 (ND10) structures but, unlike ICP0, pp71 did not induce disruption of ND10. The activity of pp71 in mouse sensory neurons in vivo was investigated after co-inoculation of animals with pairs of HSV-1 mutants, one expressing pp71 and the second containing the E. coli lacZ gene controlled by various promoters. In this system, pp71 stimulated beta-galactosidase expression from a range of viral IE promoters when mice were analysed at 4 days postinoculation. At later times, expression of pp71 resulted in a reduction in numbers of neurons containing beta-galactosidase, indicating cytotoxicity or promoter shutoff. The HSV-1 latency-active promoter was not responsive to pp71, demonstrating specificity in the activity of the protein. Pp71 was as active in mice lacking both copies of the PML gene (PML-/-) as in control animals, and in PML-/- fibroblasts pp71 stimulated gene expression as effectively as in other cell types. Therefore, neither the PML protein nor the normal ND10 structure is necessary for pp71 to stimulate gene expression. PMID- 12075079 TI - Mixed patterns of transmission of human herpesvirus-8 (Kaposi's sarcoma associated herpesvirus) in Malawian families. AB - To study transmission patterns of human herpesvirus-8 (HHV-8) (Kaposi's sarcoma associated herpesvirus) in families in Malawi, nucleotide sequences derived from two hypervariable loci of the HHV-8 genome, the V1 and V2 regions of open reading frame K1 (K1/V1 and K1/V2, respectively), were amplified from blood and mouth rinse samples of 22 patients with treated and untreated Kaposi's sarcoma (KS) and their first-degree relatives (n=67). In patients with KS, vincristine therapy was significantly associated with non-detectability of circulating, but not oral, K1/V1 DNA. Intra-familial K1/V1 phylogenetic comparisons of eight families were possible. Both identical and non-identical sequences were observed between family members, suggesting transmission of HHV-8 along both intra- and extra-familial transmission routes. PMID- 12075080 TI - Characterization of an Epstein-Barr virus-related gammaherpesvirus from common marmoset (Callithrix jacchus). AB - A gammaherpesvirus related to Epstein-Barr virus (EBV; Human herpesvirus 4) infects otherwise healthy common marmosets (Callithrix jacchus). Long-term culture of common marmoset peripheral blood lymphocytes resulted in outgrowth of spontaneously immortalized lymphoblastoid cell lines, primarily of B cell lineage. Electron microscopy of cells and supernatants showed herpesvirus particles. There were high rates of serological cross-reactivity to other herpesviruses (68-86%), but with very low geometric mean antibody titres [1:12 to human herpesvirus 6 and 1:14 to Herpesvirus papio (Cercopithecine herpesvirus 12)]. Sequence analysis of the conserved herpesvirus DNA polymerase gene showed that the virus is a member of the lymphocryptovirus subgroup and is most closely related to a lymphocryptovirus from rhesus macaques and is closely related to EBV and Herpesvirus papio. High seroprevalence (79%, with geometric mean antibody titre of 1:110) among 28 common marmosets from two geographically distinct colonies indicated that the virus is likely present in many common marmosets in captivity. A New World primate harbouring a lymphocryptovirus suggests that this subgroup arose much earlier than previously thought. PMID- 12075081 TI - Identification of structural determinants of the first transmembrane domain of the small envelope protein of duck hepatitis B virus essential for particle morphogenesis. AB - The envelope of duck hepatitis B virus (DHBV) consists of the small (S) and large (L) envelope proteins, which share a common C-terminal multispanning transmembrane region but differ by the long N-terminal pre-S domain of L, which is essential for interactions with both the receptor and nucleocapsid. To achieve these dual functions, L acquires mixed topologies through S-dependent post translational translocation of its pre-S domain. This study has examined the role of S in this unusual mechanism of translocation by analysis of the alpha-helical transmembrane domains and their potential to engage in lateral interactions for envelope assembly. Through mutagenesis in constructs expressing the S and L envelope proteins independently, transmembrane domain 1 was identified as an essential structural determinant in S. Two polar residues in this helix were identified as contributing to L protein translocation through the assembly of S into particles, implying that the topological switch of L is part of the assembly and maturation process. The same domain in L was shown to be dispensable for L translocation and assembly, suggesting that transmembrane domain 1 of L and S have different functional roles and structural arrangements on the assembled particle. The conservation in all hepadnavirus envelope proteins of two polar residues at positions 24 and 27 of transmembrane domain 1, the former positively charged, points to this being a common determinant in particle morphogenesis for all hepadnaviruses. PMID- 12075082 TI - Experimental transfection of Macaca sylvanus with cloned human hepatitis B virus. AB - Due to the absence of easily accessible animal models for the study of hepatitis B virus (HBV), the possibility of using Macaca sylvanus, a monkey originating from Morocco, North Africa, was investigated. Three monkeys were intrahepatically inoculated with a replication-competent head-to-tail HBV DNA plasmid dimer construct. The HBV surface antigen and HBV DNA were detected prior to alanine aminotransferase elevation in the serum of two of three HBV-inoculated monkeys at day 2 post-transfection and persisted for several weeks. This indicates that transfected animals developed markers of HBV infection. In addition, electron microscopy of the serum 3 weeks post-transfection showed the presence of virus particles whose shape and size were similar to complete 42 nm HBV Dane particles. Histological examination of liver tissues also revealed pathological changes not observed in uninfected controls, which strongly suggested acute hepatitis. HBV DNA was also detected by PCR in these monkey livers. Taken together, these results indicate that HBV can successfully replicate in this model and that M. sylvanus could be a potentially useful new primate model for the study of HBV replication. PMID- 12075083 TI - PTEN is a negative regulator of STAT3 activation in human papillomavirus-infected cells. AB - Laryngeal papillomas are caused by infection of the laryngeal epithelium by human papillomavirus type 6 or type 11 (HPV-6/-11). Previous studies in our laboratory have demonstrated an increase in PI3 kinase levels in papilloma tissue. However, activation of the downstream effector of PI3 kinase, protein kinase B (PKB/Akt), was reduced. This observation was explained by the elevated expression of the phosphatase and tensin homologue (PTEN), a recently characterized tumour suppressor, in papilloma tissue. Recent investigation of the possible functional roles of PTEN during papilloma development has now indicated that the level of tyrosine(705)-phosphorylated signal transducer and activator of transcription 3 [PTyr(705)STAT3] could be inversely correlated to that of PTEN as well. In vitro phosphatase assays suggested the presence of an increased level of a PTyr(705)STAT3 phosphatase in papilloma extract. Immunodepletion of PTEN from papilloma extracts resulted in a reduction of the PTyr(705)STAT3 phosphatase activity. Transfection of PTEN cDNA into HeLa cells attenuated STAT3 phosphorylation at Tyr(705) in a dose-dependent manner. This attenuation of STAT3 phosphorylation was independent of the STAT3 kinase. Interestingly, introduction of a lipid phosphatase mutant of PTEN (G129E) resulted in heightened PTyr(705)STAT3 phosphatase activity, relative to that obtained from wild-type PTEN transfection. These data indicate that PTEN negatively regulates STAT3 activation in HPV-infected papilloma cells. Induction of PTEN and reduction of activated STAT3 might be a result of a host defence mechanism or a virus-directed strategy to alter normal epithelial differentiation programming. PMID- 12075084 TI - Specific interaction of the nonstructural protein NS1 of minute virus of mice (MVM) with [ACCA](2) motifs in the centre of the right-end MVM DNA palindrome induces hairpin-primed viral DNA replication. AB - The linear single-stranded DNA genome of minute virus of mice (MVM) is replicated via a double-stranded replicative form (RF) intermediate DNA. Amplification of viral RF DNA requires the structural transition of the right-end palindrome from a linear duplex into a double-hairpin structure, which serves for the repriming of unidirectional DNA synthesis. This conformational transition was found previously to be induced by the MVM nonstructural protein NS1. Elimination of the cognate NS1-binding sites, [ACCA](2), from the central region of the right-end palindrome next to the axis of symmetry was shown to markedly reduce the efficiency of hairpin-primed DNA replication, as measured in a reconstituted in vitro replication system. Thus, [ACCA](2) sequence motifs are essential as NS1 binding elements in the context of the structural transition of the right-end MVM palindrome. PMID- 12075085 TI - Hepatitis C virus core protein expression in human B-cell lines does not significantly modify main proliferative and apoptosis pathways. AB - Hepatitis C virus (HCV) chronic infection has been associated with many lymphoproliferative disorders. Several studies performed on hepatoma and fibroblast cell lines suggest a role of the HCV core protein in activation of cellular transduction pathways that lead to cell proliferation and inhibition of apoptosis. However, no data are available concerning the effects of HCV core expression on B-lymphocyte proliferation and apoptosis. B-lymphocyte cell lines permanently expressing full-length HCV 1b core sequences isolated from chronically infected patients were established using B-cell lines at different degrees of differentiation. Clones and pools of clones permanently expressing the HCV core were selected and characterized for protein expression by Western blot and FACS. Expression of HCV core proteins did not significantly enhance cell proliferation rates under normal culture conditions or under mitogenic stimulation. Analysis of NF-kappa B, CRE, TRE and SRE pathways by luciferase reporter genes did not show a significant influence of HCV core expression on these signal transduction cascades in B-lymphocytes. The effects of HCV core on anti-IgM and anti-FAS-induced apoptosis in B-cell lines was also analysed. In this experimental model, HCV core expression did not significantly modify the apoptotic profile of the B-lymphocyte cell lines tested. These data underline a cell type-specific effect of HCV core expression. In fact, it was not possible to show a significant contribution of the HCV core protein in activation of the major B-cell signal transduction pathways involved in the regulation of proliferation and programmed cell death, which is in contrast with the results reported in hepatoma cell lines. PMID- 12075086 TI - Detection of neutralizing antibodies to hepatitis C virus using a biliary cell infection model. AB - The identification and characterization of neutralizing anti-hepatitis C virus (HCV) antibodies may have a major impact on understanding HCV pathogenesis. However, to date, their detection has only been based on the inhibition of either the E2 envelope protein or HCV virions binding to different target cells. The permissiveness of primary biliary cells for HCV infection has been demonstrated previously. In the present report, infection of biliary cells was demonstrated further by combining PCR and immunohistochemical detection of the HCV core protein. This study demonstrates, using both serum and purified IgG, the presence of neutralizing anti-HCV antibodies in the serum of patients showing long-term response to antiviral therapy. Overall, the usefulness of the primary biliary cell infection model to investigate anti-HCV neutralization is shown. PMID- 12075087 TI - Phylogenetic evidence for adaptive evolution of dengue viruses in nature. AB - A maximum-likelihood approach was used to analyse selection pressures acting on genes from all four serotypes of dengue virus (DEN). A number of amino acid positions were identified within the envelope (E) glycoprotein that have been subject to relatively weak positive selection in both DEN-3 and DEN-4, as well as in two of the five genotypes of DEN-2. No positive selection was detected in DEN 1. In accordance with the function of the E protein as the major antigenic determinant of DEN, the majority of these sites were located in, or near to, potential T- or B-cell epitopes. A smaller number of selected sites was located in other well-defined functional domains of the E protein, suggesting that cell tropism and virus-mediated membrane fusion may also confer fitness advantages to DEN in nature. Several positively selected amino acid substitutions were also identified in the NS2B and NS5 genes of DEN-2, although the cause of this selection is unclear, whereas the capsid, membrane and non-structural genes NS1, NS2A, NS3 and NS4 were all subject to strong functional constraints. Hence, evidence was found for localized adaptive evolution in natural isolates of DEN, revealing that selection pressures differ among serotypes, genotypes and viral proteins. PMID- 12075088 TI - Studies on intracellular processing of the capsid protein of human astrovirus serotype 1 in infected cells. AB - Astroviruses are non-enveloped, positive-strand RNA viruses. Their structural (capsid) protein is processed extracellularly into several smaller fragments which are found on the mature viral particle. In addition, intracellular cleavage of the capsid protein has been proposed. However, analysis of capsid protein processing has been hampered by the lack of antibodies to regions near the N and C termini of the protein. Here we describe the construction of two infectious mutants of human astrovirus serotype 1 (HAstV-1), in which amino acids (aa) 11-30 or aa 783-787, respectively, of the 787 aa capsid protein were replaced by tag sequences. Processing of the tagged capsid proteins in infected Caco-2 cells was analysed by immunoprecipitation with specific reagents directed against the tags or against native internal regions of the capsid protein. No intracellular processing of the capsid protein in infected cells could be detected, while assembled viral particles were readily observed within cells. PMID- 12075089 TI - Mapping of tissue tropism determinants in coxsackievirus genomes. AB - Genomic regions responsible for the different tissue tropisms of coxsackievirus A9 (CAV9) and coxsackievirus B3 (CBV3) in newborn mice were investigated using recombinant viruses. Infectious cDNA clones of CAV9, a virus known to infect striated muscle, and CBV3, affecting the central nervous system, pancreas, liver, brown fat and striated muscle, were used to generate chimeric viruses. In situ hybridization analysis of different tissues from mice infected with the recombinant viruses, constructed by exchanging the 5' non-coding region (5'NCR), structural and non-structural genes, demonstrated that the pancreo- and liver tropism map predominantly to CBV3 sequences within the capsid genes, evidently due to receptor recognition. Although the major neurotropism determinant in the CBV3 genome was in the capsid region, viruses containing the CAV9 capsid were also able to initiate infection in the central nervous system provided they contained the CBV3 5'NCR. The presence of the 5'NCR of CAV9 clearly enhanced muscle tissue tropism. PMID- 12075090 TI - Poliovirus pathogenesis in a new poliovirus receptor transgenic mouse model: age dependent paralysis and a mucosal route of infection. AB - We constructed a poliovirus receptor (PVR) transgenic mouse line carrying a PVR delta cDNA driven by a beta-actin promoter. We refer to this model as the cPVR mouse. The cPVR mice express Pvr in a variety of tissues (including small intestines, brain, spinal cord, muscle, blood and liver) and are susceptible to infection after intraperitoneal, intracerebral or intramuscular inoculation of poliovirus. After intraperitoneal inoculation, poliovirus replication is observed in cPVR muscle, brain, spinal cord and, notably, small intestine. The cPVR mice exhibit a striking age-dependent paralysis after intramuscular infection, with 2 week-old mice being 10,000-fold more susceptible to paralytic disease than adult mice. The cPVR mice are also susceptible to paralysis following intranasal infection with poliovirus. After intranasal infection, virus replication is observed in the olfactory bulb, cerebrum, brain stem and spinal cord, suggesting that intranasal infection of cPVR mice is a model for bulbar paralysis. Intranasally infected mice frequently display unusual neurological behaviours. The PVR transgenic mouse reported here provides the first available model for a mucosal route of infection with poliovirus. PMID- 12075091 TI - Open reading frame sequence of an Asian enterovirus 73 strain reveals that the prototype from California is recombinant. AB - Phylogenetic analysis within the VP1 region now enables molecular typing of enteroviruses consistent with neutralization results. Three untypable isolates, 2776/82, 57/99 and 22/00, from Korea, North India and Bangladesh, respectively, showed within this region 98.0-99.0% amino acid identities. These were less than 77% to the previous enterovirus prototypes, but 91.5-92.5% to CA55-1988, the recently identified enterovirus 73 (EV73) prototype from California. All three strains were, however, most similar to CA64-4454, an EV73 prime strain, to which they shared 96.5-98.5% identity. Seven compared EV73 strains formed two clusters in the VP1 dendrogram, one cluster with strains from South and East Asia and CA64 4454, and the other with strains from Oman and California including the prototype. When sequencing the complete open reading frame of 2776/82, its non structural region was found to be divergent from all human enterovirus B (HEV-B) strains, including CA55-1988, indicating that one or other strain was recombinant. Boot scanning of the genomes showed a recombination point within the P2 region. Therefore, part of this was sequenced for 57/99 and 22/00 and was found similar to 2776/82, while CA55-1988 was similar to coxsackievirus B3, demonstrating that CA55-1988 was the recombinant. Since all strains of EV73 isolated so far outside California originate from Asia, where it has a broad geographical distribution, it seems that EV73 may have been introduced to California from Asia. Further analysis of EV73 strains will reveal if the recombination occurred in the USA or in Asia and will help to elucidate the origin of this virus. PMID- 12075092 TI - Analysis of the desialidation process of the haemagglutinin protein of influenza B virus: the host-dependent desialidation step. AB - It was reported previously that haemadsorption by the haemagglutinin (HA) protein of influenza B virus required that the protein must undergo desialidation. When MDCK and COS cells were infected with influenza B/Kanagawa/73 virus in the presence of a neuraminidase (NA) inhibitor, Zanamivir, haemadsorption on MDCK cells was inhibited but that on COS cells was not. The activity of the NA protein of the two types of infected cells was similar and both were inhibited by Zanamivir in a dose-dependent manner. A comparison of the desialidation of the HA protein was made on MDCK and COS cells in the presence of bacterial NA and both cells were found to have similar sensitivity. On the accumulation of the HA and NA proteins in the trans-Golgi network of MDCK cells by means of low-temperature treatment, desialidation of the HA protein in the presence of Zanamivir was detected by two-dimensional gel electrophoresis. Because this agent was reported to be unable to penetrate cells, these data suggest that, in MDCK cells, desialidation of the HA protein occurs on the cell surface but, in COS cells, the HA and NA proteins might accumulate in the trans-Golgi network, thus allowing NA desialidation before their migration to the cell surface. PMID- 12075093 TI - Uncoupling of virus-induced inflammation and anti-viral immunity in the brain parenchyma. AB - Non-neuroadapted influenza virus confined to the brain parenchyma does not induce antigen-specific immunity. Nevertheless, infection in this site upregulated major histocompatibility complex (MHC) class I and MHC class II expression and recruited lymphocytes to a perivascular compartment. T cells recovered from the brain had an activated/memory phenotype but did not respond to viral antigens. In contrast, T cells recovered from the brain after infection in a lateral cerebral ventricle, which is immunogenic, showed virus-specific responses. As with infectious virus, influenza virus-infected dendritic cells elicited virus specific immunity when inoculated into the cerebrospinal fluid but not when inoculated into the brain parenchyma. Thus, inflammation and dendritic cell function were both uncoupled from immune priming in the microenvironment of the brain parenchyma and neither was sufficient to overcome immunological privilege. PMID- 12075094 TI - Genetic vaccines protect against Sin Nombre hantavirus challenge in the deer mouse (Peromyscus maniculatus). AB - We used a deer mouse (Peromyscus maniculatus) infection model to test the protective efficacy of genetic vaccine candidates for Sin Nombre (SN) virus that were known to provoke immunological responses in BALB/c mice (Bharadwaj et al., Vaccine 17, 2836-2843, 1999 ). Protective epitopes were localized in each of four overlapping cDNA fragments that encoded portions of the SN virus G1 glycoprotein antigen; the nucleocapsid gene also was protective. The protective efficacy of glycoprotein gene fragments correlated with splenocyte proliferation in the presence of cognate antigen, but none induced neutralizing antibodies. Genetic vaccines against SN virus can protect outbred deer mice from infection even in the absence of a neutralizing antibody response. PMID- 12075095 TI - Systemic immune responses following infection with Jaagsiekte sheep retrovirus and in the terminal stages of ovine pulmonary adenocarcinoma. AB - Jaagsiekte sheep retrovirus (JSRV) is the aetiological agent of ovine pulmonary adenocarcinoma (OPA). To monitor changes in cellular immune function during JSRV infection, lymphoproliferation in response to various mitogens was measured in the blood of conventionally housed and specific-pathogen-free lambs experimentally infected with JSRV until the development of OPA and compared with uninfected control lambs. In addition, blood samples collected from adult field cases in the terminal stages of OPA and control adult sheep were compared. No difference in the proliferative response to phytohaemagglutinin and pokeweed mitogen between the animal groups was detected. In contrast, reduced responses to concanavalin A stimulation were demonstrated in the JSRV-inoculated lambs, prior to the onset of clinical disease, and also in the terminally ill adult sheep. Peripheral blood leukocytes were monitored to identify phenotypic frequency alterations. The CD4 lymphocytopaenia and neutrophilia reported previously in adult OPA cases were demonstrated but similar phenotypic changes were not identified during experimental infection. PMID- 12075096 TI - A maedi-visna virus strain K1514 receptor gene is located in sheep chromosome 3p and the syntenic region of human chromosome 2. AB - The maedi-visna lentivirus (MVV) induces encephalitis, interstitial pneumonia, arthritis and mastitis in sheep. While some MVV strains can enter cells of ruminant species only, others can enter cells from many species, including human, but not Chinese hamster cells. However, the identity of the receptor(s) used by MVV for entry is unknown. The MVV-K1514 receptor gene was localized in sheep and human chromosomes using hamster x sheep and hamster x human hybrid cell lines. Based on entry by a vector pseudotyped with the MVV-K1514 envelope, the MVV-K1514 receptor gene was mapped to sheep chromosome 3p and to a region of human chromosome 2 (2p25>q13), which has conserved synteny with sheep chromosome 3p. These regions do not include any known lentivirus receptor or coreceptor gene, indicating that MVV-K1514 uses a new lentivirus receptor to infect human cells. PMID- 12075097 TI - Interaction between potyvirus helper component-proteinase and capsid protein in infected plants. AB - Monoclonal antibodies were raised against helper component-proteinase (HcPro) purified from plants infected with the potyvirus Lettuce mosaic virus (LMV). These antibodies were used in a two-site triple antibody sandwich ELISA assay together with polyclonal antibodies directed against purified virions. An interaction between HcPro and the viral coat protein (CP) was demonstrated in extracts of LMV-infected leaves, as well as for two other potyviruses, Plum pox virus and Potato virus Y. The CP-HcPro interaction was not abolished in LMV derivatives with an HcPro GFP N-terminal fusion, or with a deletion from the CP of the amino acids involved in aphid transmission. Electron microscopy indicated that HcPro probably does not interact with the CP in the form of assembled virions or virus-like particles. Together, these results suggest that the interaction detected between CP and HcPro might be involved in a process of the potyvirus cycle different from aphid transmission. PMID- 12075098 TI - Substitution of a single amino acid in the 2b protein of Pea early-browning virus affects nematode transmission. AB - The 2b protein of Pea early-browning virus (PEBV) is required for transmission of the virus by nematodes. Comparison of the 2b proteins of highly transmissible (TpA56) and poorly transmissible (SP5) isolates of PEBV identified two amino acid substitutions (G90S and G177R) that might be responsible for the poor transmission of isolate SP5. Hybrid viruses were created in which the TpA56 2b protein carried SP5-specific substitutions at residue 90 or 177, and in which the SP5 2b protein carried TpA56-specific substitutions at these positions. Transmission tests showed that the G177R substitution is sufficient to prevent nematode transmission of the virus. Examination of the 2b proteins from PEBV and other tobraviruses predicted the presence of a coiled-coil domain in the central region of the protein. This structural element is important for the association of interacting proteins and, thus, might mediate interaction of the 2b protein with the virus coat protein or with the vector nematode. PMID- 12075099 TI - RNA 2 of Citrus psorosis virus is of negative polarity and has a single open reading frame in its complementary strand. AB - Citrus psorosis virus (CPsV) causes a citrus disease occurring worldwide. Isolate CPV 4 has a genome with three single-stranded RNAs. The complete sequence of RNA 2 (1643 nucleotides) is reported here. Northern blot hybridization with strand specific probes showed that most of the encapsidated RNA 2 is of negative polarity, although a small amount of the complementary strand may also be present in particles. The RNA 2 complementary strand contained a single open reading frame encoding a protein of 476 amino acids, which includes a motif resembling a nuclear localization signal. The sequence of this putative protein shows no significant similarity to any other in the databases. In the 3'-terminal untranslated region there is a putative polyadenylation signal. No subgenomic RNAs derived from RNA 2 were detected. PMID- 12075100 TI - Regulated nuclear targeting of cauliflower mosaic virus. AB - The mature cauliflower mosaic virus (CaMV) capsid protein (CP), if expressed in the absence of other viral proteins, is transported into the plant cell nucleus by the action of a nuclear localization signal (NLS) close to the N terminus. In contrast, virus particles do not enter the nucleus, but dock at the nuclear membrane, a process inhibited by anti-NLS antibodies or by GTP gamma S, and apparently mediated by interaction of CP with host importin alpha. The very acidic N-terminal extension of the viral CP precursor inhibits nuclear targeting of the protein and hence the precursor is localized in the cytoplasm. We hypothesize that this provides a control mechanism which ensures that the CP precursor is used for virus assembly in the cytoplasm and that only mature virus particles reach the nuclear pore. PMID- 12075101 TI - Nucleotide sequence shows that Bean leafroll virus has a Luteovirus-like genome organization. AB - The complete nucleotide sequence of the Bean leafroll virus (BLRV) genomic RNA and the termini of its smallest subgenomic RNAs were determined to better understand its mechanisms of gene expression and replication and its phylogenetic position within the Luteoviridae: The number and placement of open reading frames (ORFs) within the BLRV genome was Luteovirus-like. The nucleotide and predicted amino acid sequences of BLRV were most similar to those of Soybean dwarf virus (SbDV). Phylogenetic analyses employing the neighbour-joining method and sister scanning analysis indicated that the BLRV nonstructural proteins were closely related to those of Barley yellow dwarf virus-PAV (BYDV-PAV), a luteovirus: The region surrounding the frameshift at the junction between ORFs 1 and 2 also contained sequences very similar to those of BYDV-PAV and a Dianthovirus, Red clover necrotic mosaic virus. Similar analyses showed that the structural proteins were most similar to those of the Polerovirus genus. The 3'-noncoding regions downstream of ORF5 contained sequences similar to translational control elements identified in the BYDV-PAV genome. These data suggest that BLRV, like SbDV, is derived either through selection from a common ancestor with BYDV-PAV or that BLRV is the product of two recombination events between luteovirus-like and polerovirus-like ancestors where the 5' 2900 nt and 3' 700 nt of the BLRV genome are from a Luteovirus and the intervening sequences are derived from a Polerovirus: PMID- 12075102 TI - Sequence analysis of Potato leafroll virus isolates reveals genetic stability, major evolutionary events and differential selection pressure between overlapping reading frame products. AB - In order to investigate the genetic diversity of Potato leafroll virus (PLRV), seven new complete genomic sequences of isolates collected worldwide were compared with the five sequences available in GenBank. Then, a restricted polymorphic region of the genome was chosen to further analyse new sequences. The sequences of PLRV open reading frames (ORFs) 3 and 4 were also compared with those of two other poleroviruses and the non-synonymous to synonymous substitution ratio distribution was analysed in overlapping and non-overlapping regions of the genome using maximum-likelihood models. Results confirmed that PLRV sequences from around the world are very closely related and showed that the region encoding protein P0 allowed the detection of three groups of isolates. When compared to other poleroviruses, PLRV was the most conserved in both ORFs 3 and 4. However, the results suggest that important events, such as deletion, mutation at a stop codon and intraspecific homologous recombination events, have occurred during the evolution of PLRV. Finally, it was shown that the translation products of ORFs 0 and 3 are significantly more conserved than those of the overlapping ORFs 1 and 4, respectively. All together, the results allow the proposal of new hypotheses to explain the apparent genetic stability of PLRV and its evolution. PMID- 12075103 TI - Testing for endocrine disruption: how much is enough? AB - The article highlighted in this issue is "Comparison of the Developmental and Reproductive Toxicity of Diethylstilbestrol Administered to Rats in Utero, Lactationally, Preweaning or from Weaning" by J. Odum, P. A. Lefevre, H. Tinwell, J. P. Van Miller, R. L. Joiner, R. E. Chapin, N. T. Wallis and J. Ashby (pp. 147 163). PMID- 12075104 TI - Approaches to assessment of the allergenic potential of novel proteins in food from genetically modified crops. AB - The safety assessment of food derived from genetically modified plants continues to attract considerable attention. Among the important issues that need to be considered is whether the products of novel genes introduced into crop plants will have the potential to induce allergic sensitization or to elicit allergic disease. Hierarchical approaches to allergenicity testing have been proposed, and these incorporate evaluation of the structural and sequence homology and serological identity of novel proteins with known allergens, measurement of resistance to proteolytic digestion, and assessment of allergenic potential using animal models. Accounts of these approaches are available elsewhere, and it is not the purpose of this article to provide a detailed critique of specific methods. Our intention is rather to look more broadly at the strategy for assessment of allergenic potential, the challenges such assessments pose for the practicing toxicologist, and how some of these might best be addressed. PMID- 12075105 TI - Ontogeny of the N-methyl-D-aspartate (NMDA) receptor system and susceptibility to neurotoxicity. AB - The NMDA receptor has been widely investigated in recent years as a target for the pharmacological management of seizures, pain and a variety of neurological disorders. Its role in normal central nervous system (CNS) activity and development, as well as in the development of CNS abnormalities and neurodegeneration has also been of interest. The NMDA receptor is one of three pharmacologically distinct subtypes of ionotropic receptor channels that are sensitive to the endogenous excitatory amino acid, L-glutamate. The ontogeny of the NMDA receptor, a multiple tetrameric and heteromeric channel complex with at least six known subunits, is controlled by three gene families and varies in developmental profile with species and regional brain area. NMDA receptors play a role in excitatory synaptic transmission, in the activity-dependent synaptic plasticity underlying learning and memory, and in pre- and postnatal CNS development, including brain cell differentiation, axonal growth and degeneration of unused neurons. The results of recent studies suggest that sustained alteration of NMDA receptor activation during critical periods of development may have deleterious effects on normal CNS development and function. Neonatal rats administered the NMDA receptor antagonists 2-amino-5-phosphonovalerate (AP5) and MK-801 during the first two weeks of life develop abnormal axonal arborization in the retinal connections to the superior colliculus, interfering with normal visual responses. Results from monkey studies suggest that chronic developmental exposure to high doses of a NMDA antagonist, remacemide, has pronounced and long lasting effects on learning. Recent findings indicate that if NMDA receptors are blocked during a specific period in neonatal life (first two weeks postnatally in the rat), massive apoptotic neurodegeneration results, due not to excitotoxic overstimulation of neurons but to deprivation of stimulation. These observations require further laboratory evidence and support in order to establish their relevance to drug-induced human neurodevelopmental concerns. It is necessary to investigate the relevance of these findings in other animal species in addition to the rat, most notably, nonhuman primates, where neuronal cytoarchitecture and development are significantly different than the rodent but more like the human. PMID- 12075106 TI - Transdermal penetration of atrazine, alachlor, and trifluralin: effect of formulation. AB - Commercial formulations of herbicides contain surfactants and other compounds to increase absorption by targeted plants. These chemicals, however, are also potential penetration enhancers for mammalian skin. The effect of formulation on dermal absorption of the herbicides atrazine, alachlor, and trifluralin and their commercial formulations Aatrex, Lasso, and Treflan was determined. In vitro absorption studies were performed by placing hairless mouse skin in a Bronough flow-through diffusion system. Donor solution was spiked with (14)C-labeled herbicide, and its penetration through the skin was monitored in 90-min fractions. Results demonstrate that dermal penetration of commercially formulated compound was significantly greater (p < 0.05) than that of the pure compound at the same concentration. The physical properties of a herbicide predicted penetration (r(2) = 0.97-0.99) for commercial formulations but were not as effective at predicting absorption for the pure compounds (r(2) = 0.51-0.71). The solvents associated with the hydrophobic herbicide Treflan altered dermal penetration of the more hydrophilic herbicides Lasso and Aatrex. Furthermore, although the most hydrophobic compound had the least penetration, it accumulated in the stratum corneum at the greatest rate. These studies can have important implications on future experiments performed to predict percutaneous penetration of herbicides. PMID- 12075107 TI - Expression and activity of microsomal epoxide hydrolase in follicles isolated from mouse ovaries. AB - Microsomal epoxide hydrolase (mEH) is involved in the detoxification of xenobiotics that are or can form epoxide metabolites, including the ovotoxicant, 4-vinylcyclohexene (VCH). This industrial chemical is bioactivated by hepatic CYP450 to the diepoxide metabolite, VCD, which destroys mouse small preantral follicles (F1). Since ovarian mEH may play a role in VCD detoxification, these studies investigated the expression and activity of mEH in isolated ovarian fractions. Mice were given 1 or 15 daily doses (ip) of VCH (7.4 mmol/kg/day) or VCD (0.57 mmol/kg/day); 4 h following the final dose, ovaries were removed, distinct populations of intact follicles (F1, 25-100 microm; F2, 100-250 microm; F3, > 250 microm) and interstitial cells (Int) were isolated, and total RNA and protein were extracted. Real-time polymerase chain reaction and the substrate cis stilbene oxide (CSO; 12.5 microM) were used to evaluate expression and specific activity of mEH, respectively. Confocal microscopy evaluated ovarian distribution of mEH protein. Expression of mRNA encoding mEH was increased in F1 (410 +/- 5% VCH; 292 +/- 5% VCD) and F2 (1379 +/- 4% VCH; 381 +/- 11% VCD) follicles following repeated dosing with VCH or VCD. Catalytic activity of mEH increased in F1 follicles following repeated dosing with VCH/VCD (381 +/- 11% VCH; 384 +/- 27% VCD). Visualized by confocal microscopy, mEH protein was distributed throughout the ovary with the greatest staining intensity in the interstitial cells and staining in the theca cells that was increased by dosing (56 +/- 0.8% VCH; 29 +/- 0.9% VCD). We conclude that mEH is expressed and is functional in mouse ovarian follicles. Additionally,in vivo dosing with VCH and VCD affects these parameters. PMID- 12075108 TI - Disposition of a low dose of bisphenol a in male and female cynomolgus monkeys. AB - Bisphenol A (BPA) is a weak estrogenic compound mass-produced with potential human exposure. Following a single oral or intravenous (iv) dose of 100 microg/kg [ring-14C(U)] radiolabeled bisphenol A (14C-BPA) to male and female cynomolgus monkeys, 79-86% of the administered radioactivity was excreted in urine over 7 days, and most of the urinary excretion was recovered by 24 h after dosing, a large part of this occurring within 12 h. The fecal excretion of radioactivity over 7 days was minimal (1.8-3.1%). Toxicokinetic parameters obtained from plasma 14C-BPA-derived radioactivity during 48 h were C(max) = 104-107 ng-eq/ml between 0.25 and 2 h, and AUC(oral) = 244-265 ng-eq*h/ml after oral dosing. In the case of the iv dose, AUC(iv) was 377-382 ng-eq*h/ml, and the bioavailability was 0.66 0.70. The terminal elimination half-life was larger post-iv dose (t(1/2iv) = 13.5 14.7 h) than post-oral dose (t(1/2oral) = 9.63-9.80 h). After iv dose, the fast phase half-life (t(1/2f)) of total radioactivity was 0.61-0.67 h. The t(1/2f) of unchanged 14C-BPA for females (0.39 h) was smaller than that for males (0.57 h). These results suggested the distribution of lipophilic 14C-BPA in adipose tissue after iv dose, in contrast to first pass metabolism after oral dose. 14C-BPA derived radioactivity was strongly bound to plasma protein (f(p) = 0.055). Radio HPLC analysis suggested the predominant plasma and urinary metabolites were mono- and diglucuronide of 14C-BPA and unchanged 14C-BPA was very low (< or =1.5%) after oral dose. These results indicate that the intestinal absorption and metabolism of BPA was rapid and extensive, and the major metabolites, glucuronide conjugates of 14C-BPA, were rapidly excreted into urine in monkeys. PMID- 12075109 TI - Mechanisms of 2-butoxyethanol carcinogenicity: studies on Syrian Hamster Embryo (SHE) cell transformation. AB - Previous studies showed that 2-butoxyethanol increased liver tumors in B6C3F1 mice following chronic exposure. While the mechanism of 2-butoxyethanol-induced liver carcinogenicity has not been defined, 2-butoxyethanol has been shown to induce hemolysis in rodents via 2-butoxyacetic acid, the major metabolite of 2 butoxyethanol. This toxic effect, coupled with the observation that continued treatment with 2-butoxyethanol results in hemosiderin deposition in the liver, has led to our hypothesis that liver carcinogenicity by 2-butoxyethnaol is mediated via oxidative stress (iron catalyzed) and Kupffer cell activation. The present study used Syrian Hamster Embryo (SHE) cell transformation, a surrogate in vitro model for carcinogenesis in vivo, to examine whether 2-butoxyethanol, 2 butoxyacetic acid, or iron (ferrous sulfate) produced cell transformation. SHE cells were treated with either 2-butoxyethanol (0.5-20 mM), 2-butoxyacetic acid (0.5-20 mM), or ferrous sulfate (0.5-75 microg/ml) for 7 days. 2-Butoxyethanol and 2-butoxyacetic acid did not induce cellular transformation. In contrast, treatment with ferrous sulfate (2.5 and 5.0 microg/ml) increased morphological transformation. Cotreatment of ferrous sulfate with the antioxidants alpha tocopherol (vitamin E) or (-)-epigallocatechin-3-gallate (EGCG) prevented ferrous sulfate-induced transformation, suggesting the involvement of oxidative stress in SHE cell transformation. The level of oxidative DNA damage (OH8dG) increased following ferrous sulfate treatment in SHE cells; additionally, using single cell gel electrophoresis (comet assay), ferrous sulfate treatment produced an increase in DNA damage. Both DNA lesions were decreased by cotreatment of ferrous sulfate with antioxidants. These data support our proposal that iron, produced indirectly through hemolysis, and not 2-butoxyethanol or its metabolite 2-butoxyacetic acid, is responsible for the observed carcinogenicity of 2-butoxyethanol. PMID- 12075110 TI - Effects of phenobarbital on DNA methylation in GC-rich regions of hepatic DNA from mice that exhibit different levels of susceptibility to liver tumorigenesis. AB - DNA methylation is an important epigenetic mechanism involved in transcriptional control and altered patterns of methylation may lead to the aberrant gene expression contributing to carcinogenesis. Three groups of mice were used in the current study: the relatively liver-tumor-sensitive C3H/He strain and B6C3F1 stock (C57BL/6xC3H/He), as well as the relatively resistant C57BL/6 strain. For a 2-week period, animals from each group were given drinking water containing a tumor-promoting dose of phenobarbital (PB), a nongenotoxic rodent carcinogen. Methylation-sensitive restriction digests using HpaII or MspI were followed by PCR amplification using an arbitrary primer or primer pair, binding preferentially to guanine and cytysine (GC)-rich regions of DNA, including CpG islands. This procedure allows for assessment of methylation at the internal and/or external cytosine of the 5'-CCGG-3' sites recognized by MspI and HpaII. Results with the single primer indicated marked differences in PB-induced hypermethylation at external and internal cytosines of 5'-CCGG-3' sites: C3H/He >> B6C3F1 > C57BL/6. Results with the arbitrary primer pair indicated PB-induced hypermethylation at the external cytosine of 5'-CCGG-3' site: B6C3F1 > C3H/He, and a low level of hypomethylation at internal and external cytosine sites in C57BL/6. Thus, there was a clear indication of more methylation changes in GC rich regions of DNA, primarily hypermethylation, in the tumor-sensitive groups of mice in response to PB treatment. Therefore, this study supports our hypothesis that the capacity to maintain normal methylation patterns is related inversely to tumor susceptibility. PMID- 12075111 TI - Acrylonitrile is a multisite carcinogen in male and female B6C3F1 mice. AB - Acrylonitrile is a heavily produced unsaturated nitrile, which is used in the production of synthetic fibers, plastics, resins, and rubber. Acrylonitrile is a multisite carcinogen in rats after exposure via gavage, drinking water, or inhalation. No carcinogenicity studies of acrylonitrile in a second animal species were available. The current studies were designed to assess the carcinogenicity of acrylonitrile in B6C3F1 mice of both sexes. Acrylonitrile was administered by gavage at 0, 2.5, 10, or 20 mg/kg/day, 5 days per week, for 2 years. Urinary thiocyanate and N-acetyl-S-(2-cyanoethyl)-L-cysteine were measured as markers of exposure to acrylonitrile. In general, there were dose-related increases in urinary thiocyanate and N-acetyl-S-(2-cyanoethyl)-L-cysteine concentrations in all dosed groups of mice and at all time points. Survival was significantly (p < 0.001) reduced in the top dose (20 mg/kg) group of male and female mice relative to controls. The incidence of forestomach papillomas and carcinomas was increased in mice of both sexes in association with an increase in forestomach epithelial hyperplasia. The incidence of Harderian gland adenomas and carcinomas was also markedly increased in the acrylonitrile-dosed groups. In female mice, the incidence of benign or malignant granulosa cell tumors (combined) in the ovary in the 10 mg/kg dose group was greater than that in the vehicle control group, but because of a lack of dose response, this was considered an equivocal finding. In addition, the incidences of atrophy and cysts in the ovary of the 10 and 20 mg/kg dose groups were significantly increased. The incidences of alveolar/bronchiolar adenoma or carcinoma (combined) were significantly increased in female mice treated with acrylonitrile at 10 mg/kg/day for 2 years. This was also considered an equivocal result. In conclusion, these studies demonstrated that acrylonitrile causes multiple carcinogenic effects after gavage administration to male and female B6C3F1 mice for 2 years. PMID- 12075112 TI - Cytochrome P4501A (CYP1A) in killifish (Fundulus heteroclitus): heritability of altered expression and relationship to survival in contaminated sediments. AB - Previous research has shown that killifish (Fundulus heteroclitus) inhabiting a creosote-contaminated site on the Elizabeth River in Virginia exhibit little induction of cytochrome P4501A (CYP1A) protein expression and activity upon exposure to typical CYP1A-inducing chemicals. We characterized the CYP1A response of first, second, and third generation laboratory-raised offspring of feral Elizabeth River killifish to exposure to sediments from the contaminated site as well as the prototypical polycyclic aromatic hydrocarbon (PAH)-type CYP1A inducers beta-naphthoflavone (BNF) and 3-methylcholanthrene (3-MC). The Elizabeth River offspring's responses were compared to those of offspring of killifish from two reference sites (King's Creek, Virginia, and Russell Creek, North Carolina). As with feral Elizabeth River killifish, the first generation embryos and larvae were refractory to CYP1A induction. However, the response observed in 3-year-old first generation adults, as well as with second and third generation fish, was much closer to that observed in reference-site fish. We suggest that the pattern of altered CYP1A response in Elizabeth River killifish, while persistent and heritable for one generation, is mostly nongenetically based. Additionally, we investigated the hypothesis that low CYP1A activity (measured as in ovo EROD activity) would correlate to increased survival in Elizabeth River sediment pore water; this hypothesis was not supported by our results. PMID- 12075113 TI - Differential activation of the c-Jun N-terminal kinase pathway in arsenite induced apoptosis and sensitization of chemically resistant compared to susceptible B-lymphoma cell lines. AB - Various forms of inorganic arsenic are significant environmental contaminants that have multiple effects on cells, including the induction of apoptotic cell death. Induction of apoptosis in lymphoid cells can mediate immunotoxicity following exposure to chemicals. However, the mechanisms regulating the sensitivity of B-lymphocytes to arsenic-induced apoptosis are not understood. Therefore, we investigated the involvement of key mitogen-activated protein kinase pathways and apoptosis induction by sodium arsenite in a model system of chemically resistant and susceptible B-lymphoma cell lines. These studies revealed a differential requirement for the c-Jun N-terminal kinase (JNK) pathway for apoptosis induction by sodium arsenite in the resistant EW36 versus sensitive ST486 cell lines. Specifically, activation of the JNK pathway was not required for arsenite-induced apoptosis in ST486 cells, whereas JNK pathway activation was always associated with apoptosis induction in EW36 cells. Importantly, we found that EW36 cells, which overexpress the Bcl-2 protein, can be substantially sensitized to arsenite-induced apoptosis by prior exposure to nonlethal hyperthermia. Moreover, pretreatment with an inhibitor of p38 kinase acted synergistically with hyperthermia to further sensitize EW36 cells. The inhibition of p38 prolonged a transient period of JNK phosphorylation that occurred immediately after heat shock treatment and involved the persistent activation of SEK1, one of the kinases upstream of JNK. Significantly, the sensitization of resistant cells is characterized by a lowering of the threshold concentration of arsenite required to activate the JNK pathway and induce apoptosis. PMID- 12075114 TI - Etomoxir-induced oxidative stress in HepG2 cells detected by differential gene expression is confirmed biochemically. AB - Although they are known to be effective antidiabetic agents, little is published about the toxic effects of carnitine palmitoyltransferase-1 (CPT-1) inhibitors, such as etomoxir (ET). These compounds inhibit mitochondrial fatty acid beta oxidation by irreversibly binding to CPT-1 and preventing entry of long chain fatty acids into the mitochondrial matrix. Treatment of HepG2 cells with 1 mM etomoxir for 6 h caused significant modulations in the expression of several redox-related and cell cycle mRNAs as measured by microarray analysis. Upregulated mRNAs included heme oxygenase 1 (HO1), 8-oxoguanine DNA glycosylase 1 (OGG1), glutathione reductase (GSR), cyclin-dependent kinase inhibitor 1A (CDKN1 [p21(waf1)]) and Mn+ superoxide dismutase precursor (SOD2); while cytochrome P450 1A1 (CYP1A1) and heat shock 70kD protein 1 (HSPA1A) were downregulated. Real time quantitative PCR (RT-PCR) confirmed the significant changes in 4 of 4 mRNAs assayed (CYP1A1, HO1, GSR, CDKN1), and identified 3 additional mRNA changes; 2 redox-related genes, gamma-glutamate-cysteine ligase modifier subunit (GCLM) and thioredoxin reductase (TXNRD1) and 1 DNA replication gene, topoisomerase IIalpha (TOP2A). Temporal changes in selected mRNA levels were examined by RT-PCR over 11 time points from 15 min to 24 h postdosing. CYP1A1 exhibited a 38-fold decrease by 4 h, which rebounded to a 39-fold increase by 20 h. GCLM and TXNRD1 exhibited 13- and 9-fold increases, respectively at 24 h. Etomoxir-induced oxidative stress and impaired mitochondrial energy metabolism were confirmed by a significant decrease in reduced glutathione (GSH), reduced/oxidized glutathione ratio (GSH/GSSG), mitochondrial membrane potential (MMP), and ATP levels, and by concurrent increase in oxidized glutathione (GSSG) and superoxide generation. This is the first report of oxidative stress caused by etomoxir. PMID- 12075115 TI - Gene-specific TCDD suppression of RARalpha- and RXR-mediated induction of tissue transglutaminase. AB - The malignant human keratinocyte line SCC4 provides a model system to study the mechanism by which 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) suppresses retinoid induction of the tissue transglutaminase gene (TGM2). The current work explores the nature of TCDD suppression of retinoid action to determine whether it is gene specific, whether it is retinoid receptor isoform-dependent, and whether it requires close proximity of retinoid and TCDD response elements. First, two other retinoid-inducible genes were identified in SCC4 by microarray screening whose induction was unaffected by TCDD, clearly demonstrating the gene specificity of TCDD suppression. Second, the receptor isoform dependence of retinoid responsiveness in SCC4 was tested. TGM2 was found to be inducible by an RARalpha specific but not by an RARgamma-selective agonist. A lack of responsiveness to RARgamma agonists was found to be characteristic of SCC4, however, inasmuch as transcription driven by a retinoid response element in transfections was also stimulated only by the alpha-specific agonist in these cells. Because SCC4 lacks expression of RARbeta, the gene specificity evidently was not attributable to differential TCDD targeting of retinoid receptor isoforms. Finally, the proximal 5 kb of the TGM2 promoter was found to be retinoid responsive in stable transfections, but the induction was not suppressed by TCDD. These results indicate that the suppressive action of TCDD occurs indirectly and through a separate DNA site likely located outside the 5-kb region, not by direct interference with retinoid action or at retinoid response elements. PMID- 12075116 TI - Neurobehavioral assessments of rats perinatally exposed to a commercial mixture of polychlorinated biphenyls. AB - Because of behavioral deficits associated with gestational exposure to PCBs in children, we sought to quantify neurobehavioral effects of perinatal exposure to Aroclor 1254(R) (A1254), a commercial mixture of PCBs, in rats. Pregnant Long Evans rats were fed A1254 at doses of 0, 1.0, or 6.0 mg/kg/day throughout gestation and nursing. The growth and behavior of their male and female offspring were assessed both during development and as adults, using a variety of behavioral tests that included a neurobehavioral screening battery (functional observational battery [FOB] and automated tests of locomotor activity), habituation of motor activity, acquisition of a visual discrimination, and performance of a visual signal-detection task. During the suckling period, A1254 at 6 mg/kg reduced survival and body weight gain of offspring of both sexes; however, locomotor activity was unaffected, and only small and transient changes in other measures were evident. In adulthood, perinatal exposure to A1254 did not affect habituation of locomotor activity, acquisition of the visual discrimination, or sustained attention. Rats performing the signal-detection task were challenged with cocaine (0, 1.25, 2.5, 5.0 mg/kg) and haloperidol (0, 0.003, 0.010, 0.030 mg/kg) to probe the integrity of dopaminergic systems in the central nervous system (CNS). A1254 did not alter the impairment of attention caused by haloperidol. Cocaine reduced false alarms more in controls than in rats exposed to A1254, but the effect was not clearly related to the dose of A1254. Perinatal exposure to this commercial PCB mixture had very little effect on these tests of behavior during development and in adulthood. PMID- 12075117 TI - Three-generation reproductive toxicity study of dietary bisphenol A in CD Sprague Dawley rats. AB - Bisphenol A (BPA) was evaluated at concentrations of 0, 0.015, 0.3, 4.5, 75, 750, and 7500 ppm ( approximately 0.001, 0.02, 0.3, 5, 50, and 500 mg/kg/day of BPA) administered in the diet ad libitum to 30 CD((R)) Sprague-Dawley rats/sex/dose for 3 offspring generations, 1 litter/generation, through F3 adults. Adult systemic toxicity at 750 and 7500 ppm in all generations included: reduced body weights and body weight gains, reduced absolute and increased relative weanling and adult organ weights (liver, kidneys, adrenals, spleen, pituitary, and brain), and female slight/mild renal and hepatic pathology at 7500 ppm. Reproductive organ histopathology and function were unaffected. Ovarian weights as well as total pups and live pups/litter on postnatal day (PND) 0 were decreased at 7500 ppm, which exceeded the adult maximum tolerated dose (MTD). Mating, fertility, gestational indices; ovarian primordial follicle counts; estrous cyclicity; precoital interval; gestational length; offspring sex ratios; postnatal survival; nipple/areolae retention in preweanling males; epididymal sperm number, motility, morphology; daily sperm production (DSP), and efficiency of DSP were all unaffected. At 7500 ppm, vaginal patency (VP) and preputial separation (PPS) were delayed in F1, F2, and F3 offspring, associated with reduced body weights. Anogenital distance (AGD) on PND 0 was unaffected for F2 and F3 males and F3 females (F2 female AGD was increased at some doses, not at 7500 ppm, and was considered not biologically or toxicologically relevant). Adult systemic no observed adverse effect level (NOAEL) = 75 ppm (5 mg/kg/day); reproductive and postnatal NOAELs = 750 ppm (50 mg/kg/day). There were no treatment-related effects in the low-dose region (0.001-5 mg/kg/day) on any parameters and no evidence of nonmonotonic dose-response curves across generations for either sex. BPA should not be considered a selective reproductive toxicant, based on the results of this study. PMID- 12075118 TI - Comparison of the developmental and reproductive toxicity of diethylstilbestrol administered to rats in utero, lactationally, preweaning, or postweaning. AB - The objective of the study was to determine which period of exposure produces the most marked effects on the reproductive capacity and sexual development of the rat, with particular emphasis on the relative sensitivity of in utero and postnatal exposures. The endocrine active chemical, diethylstilbestrol (DES) was used as an agent known to affect many of the endpoints examined. Hitherto, such comparisons have been made between studies, rather than within a study. Our data will be helpful in the interpretation of future multigenerational assay data. In preliminary studies, DES was shown to be active in the immature rat uterotrophic assay with a lowest detected dose of 0.05 mg DES/kg body weight by sc injection and 10 mg DES/l (1.6 mg DES/kg body weight) by administration in drinking water. A dose of 60 microg DES/l drinking water ( approximately 6.5mg DES/kg body weight/day) was selected for the main study since this represented the midpoint of the drinking water uterotrophic dose response and produced no overt maternal toxicity. The study used 10 groups of concomitantly pregnant animals, including 2 control groups. The first comparison was between the effects of exposure to DES in utero, and exposure from conception to weaning. Another group of animals was exposed to DES in utero and cross-fostered to untreated pregnant females to prevent lactational transfer of DES to pups. Two groups were exposed to DES neonatally, either from birth to postnatal day (PND) 10 (pups thus having only lactational exposure), or from birth until weaning (PND 21; pups thus having both lactational exposure and self-exposure via drinking water). In addition, a dose response study to DES was conducted on animals exposed from weaning to PND 100, when the first phase of the study was terminated. Pups exposed to DES in utero and pups exposed from weaning to PND 100 were bred to assess fertility of the F1 animals and the sexual development of F2 offspring. This last comparison was to determine the extent to which weanling rats could be used in endocrine toxicity studies to assess their potential to show activity in utero. The most sensitive period of exposure for inducing developmental effects in F1 animals was from weaning onwards. The neonatal to weaning period (PND 1-21) was the next most sensitive. Essentially no effects were induced in F1 animals exposed in utero. No effects of any kind were observed in animals only exposed over the early neonatal period of PND 1-10. The mean day of vaginal opening, testes descent, and prepuce separation was only altered in groups where postnatal exposure to DES continued beyond PND 10, or was started at weaning. No changes were observed in anogenital distance or caudal sperm counts. Some changes in organ weights were observed, but the interpretation of these was often confused by concomitant changes in body weight. In general, histopathological examination of tissues yielded no additional information. In breeding studies with animals exposed to DES in utero, or from weaning, reduced litter sizes and marginal advances in the day of vaginal opening were observed in the offspring, together with changes in organ weights. However, no unique sensitivity was noted for exposure in utero. Evaluation of the several exposure periods and the many markers monitored in this study may have individual strengths in individual cases, but when rigorously compared using the reference estrogen DES, many preconceptions regarding their absolute or relative value were not upheld. Further, each of these markers is subject to natural variability, as demonstrated by comparisons made among the 5 separate control groups available in parts of the present study. This variability increases the chance that small changes observed in endocrine toxicity studies employing small group sizes and a single control group, or no concomitant control group, may be artifactual. The most marked effects observed in this study were on the developmental landmarks in the F1 animals induced by exposures after PND 10. Some effects on developmental landmarks and organ weights were observed in F2 animals following exposure either in utero or postweaning. This study therefore does not establish a unique role for exposures in utero or during the early neonatal period. PMID- 12075119 TI - Bromochloroacetic acid exerts qualitative effects on rat sperm: implications for a novel biomarker. AB - Disubstituted haloacid by-products of drinking water disinfection such as dibromoacetic acid and dichloroacetic acid have been shown to perturb spermatogenesis and fertility in adult male rats. In the present study we sought to establish whether equimolar exposure to bromochloroacetic acid (BCA), a prevalent by-product in finished drinking water, is also capable of disrupting these endpoints, and if so to determine whether the novel biomarker of fertility (SP22) would be correlated with subfertility induced by testicular toxicity. A dose range finding study indicated that body weight was not affected by exposure to 14 daily doses of 72 mg/kg BCA while numerous male reproductive parameters were altered, including decreases in the number and progressive motility of cauda epididymal sperm. In addition, there was an increased incidence of delayed spermiation in the testes of males exposed to 72 mg/kg BCA. In the definitive study, exposures ranged from 8 to 72 mg/kg, the fertility of cauda epididymal sperm was evaluated by in utero insemination, and the two-dimensional profile of cauda sperm membrane proteins was evaluated quantitatively. The morphology of both caput and cauda epididymal sperm was altered by 72 mg/kg BCA. The fertility of cauda epididymal sperm, the percentages of progressively motile sperm and progressive tracks, and two sperm membrane proteins (SP22 and SP9) were decreased significantly by each BCA exposure. While the two sperm proteins and the two measures of progressive motility were each significantly correlated with fertility, only one of these measures (i.e., SP22) had an r value of greater than 0.5. When data for SP22 and fertility were fit to a nonlinear model, r(2) was 0.84. Using this exposure paradigm, the no-observed-effect level for BCA is less than 8 mg/kg. Moreover, SP22 may be useful in predicting compromised fertility after exposure to by-products of drinking water disinfection. PMID- 12075120 TI - Effects of in utero exposure to the organophosphate insecticide fenitrothion on androgen-dependent reproductive development in the Crl:CD(SD)BR rat. AB - Fenitrothion [0,0-dimethyl-O-(4-nitro-m-tolyl) phosphorothioate] is an organophosphate insecticide that has been shown to have antiandrogenic activity using in vitro and in vivo screening assays. Studies were performed to evaluate the ability of fenitrothion to disrupt androgen-dependent sexual differentiation in the male rat. Pregnant Crl:CD(SD)BR rats were administered fenitrothion by gavage at 0, 5, 10, 15, 20, or 25 mg/kg/day ( n = 6-11/group) from gestation day (GD) 12 to 21. Maternal toxicity was observed in the dams treated with 20 and 25 mg fenitrothion/kg/day based on muscle tremors and decreases in body weight gain from GD 12 to 21. Fetal death was increased in the 20 and 25 mg/kg/day exposure groups, as evidenced by a decrease in the proportion of pups born alive. Androgen mediated development of the reproductive tract was altered in male offspring exposed in utero to maternally toxic levels of fenitrothion (25 mg/kg/day), as evidenced by reduction in anogenital distance on postnatal day (PND) 1 and retention of areolae on PND 13. However, these effects were only transient, and there were no indications of abnormal phenotypes or development of androgen dependent tissues on PND 100. At the dose levels evaluated in this study, fenitrothion was only weakly antiandrogenic in vivo compared with other androgen receptor antagonists such as flutamide, linuron, and vinclozolin. Based on observed fetotoxicity at 20 mg/kg/day, the lowest observed adverse effect level (LOAEL) for developmental effects can be lowered from 25 to 20 mg/kg/day. PMID- 12075121 TI - Gene expression profile induced by 17alpha-ethynyl estradiol, bisphenol A, and genistein in the developing female reproductive system of the rat. AB - Exposure to some compounds with estrogenic activity, during fetal development, has been shown to alter development of reproductive organs, leading to abnormal function and disease either after birth or during adulthood. In order to understand the molecular events associated with the estrogenicity of different chemicals and to determine whether common sets of gene expression changes can be predictive of estrogenic activity, we have used microarray technology to determine the transcriptional program influenced by exposure to this class of compounds during organogenesis and development. Changes in patterns of gene expression were determined in the developing uterus and ovaries of Sprague-Dawley rats on GD 20, exposed to graded dosages (sc) of 17alpha-ethynyl estradiol (EE), genistein, or bisphenol A (BPA) from GD 11 to GD 20. Dose levels were roughly equipotent in estrogenic activity. We compared the transcript profiles between treatment groups and controls, using oligonucleotide arrays to determine the expression level of approximately 7000 rat genes and over 1000 expressed squence tags (ESTs). At the highest tested doses of EE, BPA, or genistein, we determined that less than 2% of the mRNA detected by the array showed a 2-fold or greater change in their expression level (increase or decrease). A dose-dependent analysis of the transcript profile revealed a common set of genes whose expression is significantly and reproducibly modified in the same way by each of the 3 chemicals tested. Additionally, each compound induces changes in the expression of other transcripts that are not in common with the others, which indicated not all compounds with estrogenic activity act alike. The results of this study demonstrate that transplacental exposure to chemicals with estrogenic activity changes the gene expression profile of estrogen-sensitive tissues, and that the analysis of the transcript profile of these tissues could be a valuable approach to determining the estrogenicity of different compounds. PMID- 12075122 TI - A study of the potential for a herbicide formulation containing 2,4-d and picloram to cause male-mediated developmental toxicity in rats. AB - Male Vietnam veterans have repeatedly expressed concern that exposure to herbicides in Vietnam may have caused birth defects in their offspring. The second most used herbicide was a mixture of 2,4-D and picloram called Agent White. This study is an investigation into the possible male-mediated reproductive toxicology of this herbicide. Male rats were gavaged for 5 days per week for 9 weeks with a mixture of 2,4-D and picloram called Tordon 75D(R) (the Australian derivative of Agent White). Three doses were tested; the high dose was considered the maximum tolerated dose. Each male was mated with two untreated females during weeks 2 and 3, 4 and 5, and 8 and 9 of treatment, and with four untreated females after an 11-week recovery period. Negative controls were males dosed with distilled water, and positive controls were males dosed with cyclophosphamide at 5.1 mg/kg/day. All mated females were killed on day 20 of gestation, and the fetuses were weighed and examined for either structural malformations or skeletal development. Litter size, fetal weight, and malformation rate were all unaffected by treatment. The cyclophosphamide positive controls showed the expected large increase in postimplantation loss. In general, within the limitations of the power of the study, the results did not show any evidence that exposure to a herbicide formulation containing 2,4-D and picloram is likely to cause male-mediated birth defects or other adverse reproductive outcomes. PMID- 12075123 TI - Embryo-fetal developmental and reproductive toxicology of vinyl chloride in rats. AB - Vinyl chloride (VC) exposure is primarily via inhalation in the workplace. The primary target organ of VC toxicity is the liver and occupational exposure to VC leads to hepatic angiosarcoma. However, based on epidemiological studies, researchers have been unable to ascertain the effect of occupational VC exposure on embryo-fetal development or reproductive function. A limited number of animal studies available in the literature have examined the effect of VC on embryo fetal development, however, there are no published studies on the effect of VC exposure on reproductive capability. The current study was designed to assess the potential maternal and/or embryo-fetal developmental and 2-generation reproductive toxicity of inhaled VC in CD(R) Sprague-Dawley rats at exposure levels of 0, 10, 100, and 1100 ppm. In the embryo-fetal/developmental toxicity study, the female rats were exposed to VC daily from gestation day (GD) 6 through 19. In the reproductive toxicity study, the F(0) generation male and female rats were exposed to VC for a 10-week premating and 3-week mating periods. The F(0) generation male rats were exposed to VC until terminal euthanasia. The F(0) generation female rats were exposed from GD 0 through GD 20 and lactation day (LD) 4 through LD 25. Our results indicate that up to 1100 ppm VC exposure did not adversely affect embryo-fetal developmental or reproductive capability over 2 generations in rats. The primary target organ of VC, the liver, was affected as evidenced by an increase in liver weight and/or histologically identified cellular alterations, such as centrilobular hypertrophy at 100 and 1000 ppm. Based on the results of these studies, the no observed adverse effect level (NOAEL) for embryo-fetal/development is 1100 ppm, and the NOAEL for reproduction is 1100 ppm. The results from the current studies, which are a more comprehensive embryo-fetal/developmental and reproduction study, may be incorporated into future risk assessments of occupational exposure to VC where concerns regarding the effects of VC exposure remain. PMID- 12075124 TI - Bacterial lipopolysaccharide exposure alters aflatoxin B(1) hepatotoxicity: benchmark dose analysis for markers of liver injury. AB - Aflatoxin B(1) (AFB(1)) is a fungal toxin that causes both acute hepatotoxicity and hepatocellular carcinoma in humans and experimental animals. Previous studies demonstrated that a small, noninjurious dose of bacterial lipopolysaccharide (LPS) augments the hepatotoxicity of AFB(1) through activation of inflammatory cells and production of soluble inflammatory mediators (Barton et al., 2000b, 2001). This study was conducted to examine the effect of LPS on the dose-response relationship for AFB(1)-induced liver injury. Male Sprague-Dawley rats (250-350g) were treated with AFB(1) (0.1 mg/kg-6.3 mg/kg, ip) and 4 h later with a noninjurious dose of E. coli LPS (7.4 x 10(6) EU/kg, iv). Twenty-four h after AFB(1) administration, hepatic parenchymal cell injury was estimated from elevations in serum alanine aminotransferase and aspartate aminotransferase activities. Injury to intrahepatic bile ducts was evaluated from increased serum gamma-glutamyl transferase and alkaline phosphatase activities. Based on benchmark dose (BMD) analysis, the AFB(1) BMD for parenchymal cell injury was decreased 10-fold by LPS cotreatment, whereas AFB(1) BMDs for bile duct injury were decreased nearly 20-fold. The data suggest that concurrent inflammation renders the liver considerably more sensitive to the hepatotoxic effects of AFB(1). PMID- 12075125 TI - Rodent carcinogenicity with the thiazolidinedione antidiabetic agent troglitazone. AB - Carcinogenic potential of the thiazolidinedione antidiabetic troglitazone was assessed in 104-week studies in mice and rats. Mice were given 50, 400, or 800 mg/kg, male rats 100, 400, or 800 mg/kg, and female rats 25, 50, or 200 mg/kg. Vehicle and placebo controls were included. Survival was significantly decreased in both sexes of both species at high doses, but was adequate for valid evaluation of carcinogenicity. Hypertrophy and hyperplasia of brown adipose tissue was observed in both species at all doses, and fatty change and hypocellularity of bone marrow was noted in mice at all doses and in female rats at 50 and 200 mg/kg. Hepatocellular vacuolation was observed in mice at 400 and 800 mg/kg, and centrilobular hepatocellular hypertrophy occurred in rats at > or = 200 mg/kg. Ventricular dilatation, myocardial fibrosis, and atrial myocyte karyomegaly in male rats at 400 and 800 mg/kg and female rats at all doses were morphologically similar to spontaneous lesions, but incidence and severity were increased compared with controls. In mice, the incidence of hemangiosarcoma was increased in females at 400 mg/kg and in both sexes at 800 mg/kg. The incidence of hepatocellular carcinoma was increased in female mice at 800 mg/kg. Troglitazone exposure [AUC((0-24))] at the lowest dose associated with increased tumor incidence in mice was 16 times human therapeutic exposure at 400 mg daily. No tumors of any type were increased in rats at exposures up to 47 times therapeutic exposure. PMID- 12075126 TI - Mainstream and sidestream cigarette smoke inhibit growth and angiogenesis in the day 5 chick chorioallantoic membrane. AB - The purpose of this study was to test the hypothesis that components in mainstream (MS) and sidestream (SS) cigarette smoke inhibit growth and angiogenesis using the chick chorioallantoic membrane (CAM). Varying doses of whole or gas-phase MS and SS smoke solutions were placed on day 5 CAMs, and their effects on angiogenesis were evaluated on day 6. All parameters evaluated (CAM area, major blood vessel area, major blood vessel diameter, blood vessel pattern formation, and capillary plexus formation) were inhibited to different degrees in a dose-dependent manner by both MS and SS smoke treatment. Inhibition of growth and vessel development was correlated with inhibition of cell proliferation. Inhibition of capillary plexus formation was caused by failure of mesodermal blood vessels to migrate to the ectoderm. SS smoke solution was more inhibitory than MS smoke solution in all assays, except for capillary plexus formation. In all assays, the toxicants in SS smoke partitioned mainly with the gas phase, whereas those in MS smoke were deduced to be mainly in the particulate phase in the proliferation-dependent assays (CAM area, blood vessel area, blood vessel diameter) and in both the gas and particulate phase in the pattern formation and plexus formation assays. Some of the inhibitory doses of MS and SS smoke solutions had nicotine concentrations within the range found in human smokers. Taken together, these data demonstrate that exposure to complex mixtures of chemicals in MS and SS cigarette smoke adversely affect growth, vessel development, vessel migration, and cell proliferation. PMID- 12075127 TI - Subchronic toxicity studies on perfluorooctanesulfonate potassium salt in cynomolgus monkeys. AB - This study was conducted to determine the earliest measurable response of primates to low-level perfluorooctanesulfonate (PFOS) exposure and to provide information to reduce uncertainty in human health risk assessment. Groups of male and female monkeys received 0, 0.03, 0.15, or 0.75 mg/kg/day potassium PFOS orally for 182 days. Recovery animals from each group, except the 0.03 mg/kg/day dose group, were monitored for one year after treatment. Significant adverse effects occurred only in the 0.75 mg/kg/day dose group and included compound related mortality in 2 of 6 male monkeys, decreased body weights, increased liver weights, lowered serum total cholesterol, lowered triiodothyronine concentrations (without evidence of hypothyroidism), and lowered estradiol levels. Decreased serum total cholesterol occurred in the 0.75 mg/kg/day dose group at serum PFOS levels > 100 ppm. Hepatocellular hypertrophy and lipid vacuolation were present at term in the 0.75 mg/kg/day dose group. No peroxisomal (palmitoyl CoA oxidase) or cell proliferation (proliferating cell nuclear antigen immunohistochemistry) was detected. Complete reversal of clinical and hepatic effects and significant decreases in serum and liver PFOS occurred within 211 days posttreatment. Liver to-serum PFOS ratios were comparable in all dose groups, with a range of 1:1 to 2:1. Serum concentrations associated with no adverse effects (0.15 mg/kg/day) were 82.6 +/- 25.2 ppm for males and 66.8 +/- 10.8 ppm for females. Comparison of serum PFOS concentrations associated with no adverse effect in this study to those reported in human blood samples (0.028 +/- 0.014 ppm) indicated an adequate margin of safety. PMID- 12075128 TI - Bolbophorus damnificus n. sp. (Digenea: Bolbophoridae) from the channel catfish Ictalurus punctatus and American white pelican Pelecanus erythrorhynchos in the USA based on life-cycle and molecular data. AB - The common pathogenic prodiplostomulum metacercaria in the flesh, mostly near the skin, of pond-produced channel catfish Ictalurus punctatus has been demonstrated to be Bolbophorus damnificus Overstreet & Curran n. sp. The catfish acquires the infection from the snail Planorbella trivolvis, the only known first intermediate host, and the species is perpetuated through the American white pelican Pelecanus erythrorhynchos, as confirmed by experimental infections with nestling and dewormed adult pelican specimens in conjunction with molecular data. It differs from the cryptic species Bolbophorus sp., also found concurrently in the American white pelican, by having eggs 123-129 microm rather than 100-112 microm long and consistent low values for nucleotide percentage sequence similarity comparing COI, ITS 1/2, 18S rRNA and 28S rRNA fragments. Bolbophorus sp. is comparable but most likely distinct from B. confusus (Kraus, 1914), which occurs in Europe and has eggs 90-102 microm long. Its intermediate hosts were not demonstrated. The adults of neither of the confirmed North American species of Bolbophorus were encountered in any bird other than a pelican, although several shore birds feed on infected catfish, and B. damnificus can survive but not mature when protected in the mouse abdominal cavity. B. ictaluri (Haderlie, 1953) Overstreet & Curran n. comb., a species different from B. damnificus, is considered a species inquirenda. PMID- 12075129 TI - Testicular myxosporidiasis in anurans, with a description of Myxobolus fallax n. sp. AB - During studies of amphibian sperm cryopreservation, a new species of myxosporidean parasite (Myxozoa, Myxosporae) was observed in the testes of the Australian dwarf green tree frog Litoria fallax (Peters). Myxosporidiasis was found to have no affect on L. fallax body condition or sperm numbers. Myxobolus spores from L. fallax are morphologically distinct from Myxobolus hylae spores (infecting the sympatric Litoria aurea Lesson) and the three previously named (exotic to Australia) Myxobolus species found in anurans. Myxobolus fallax n. sp. is characterised by: pseudocyst white, spherical to ovoid, 141 x 74 to 438 x 337 microm in diameter (mature); plasmodium with spores loosely arranged within interior. Spores ovoid 13.4 +/- 0.5 (12.6-14.6) microm length, 9.5 +/- 0.4 (8.3 10.6) microm width, 6.8 +/- 0.4 (6.5-7.6) microm depth, 1.4 +/- 0.1 (1.3-1.6) length/width; polar capsules broadly pyriform and equal in size 4.2 +/- 0.3 (3.3 4.7) microm length, 2.4 +/- 0.2 (2.1-2.8) microm width; filament coils 7-8, wound tightly and perpendicular to the longitudinal axis of the capsule; polar filament 34 +/- 7.0 (18-50) microm length; intercapsular appendix and sutural ridge folds absent; and iodinophilous vacuole and mucous envelope lacking. In addition to this new species, data from archival samples of M. hylae are provided which show two morphologically distinct spore types. Both appeared rarely in the same pseudocysts and we cautiously retain the single species. PMID- 12075130 TI - The genus Biuterina Fuhrmann, 1902 (Cestoda, Paruterinidae) in the Old World: redescriptions of four species from Afrotropical passeriformes. AB - Four species of Biuterina Fuhrmann, 1902 (Cestoda, Cyclophyllidea, Paruterinidae), originally described from Afrotropical passeriform birds, are redescribed and figured on the basis of their type-specimens. These are B. pentamyzos (Mettrick, 1960) from Prionops plumata (Laniidae) in Zimbabwe, B. quelea (Mettrick, 1963) from Quelea quelea (Ploceidae) in Zimbabwe, B. ugandae Baylis, 1919 from Chalcomitra senegalensis (Nectariniidae) in Uganda and B. zambiensis (Mettrick, 1960) from Campephaga flava (Campephagidae) in Zimbabwe. The rostellum of B. quelea is a spherical structure filled with strongly developed glandular tissue and possessing a weak musculature. The remaining three species have a sucker-like rostellar apparatus with moderately developed glandular tissue in the rostellum and around it. It is considered that the glandular elements are an inherent part of the rostellar apparatus of the paruterinids. An armament consisting of fine, punctiform, spine-like structures arranged in transverse rows on the inner surface of suckers is observed in B. pentamyzos. This is the first record of sucker armature in the Paruterinidae. PMID- 12075131 TI - Litomosoides anguyai n. sp. (Nematoda: Onchocercidae) from Oxymycterus misionalis (Rodentia: Muridae) in the rain forest of Misiones, Argentina. AB - A new species of Litomosoides is described from sigmodontine murids occurring in the rain forests of Misiones, Argentina. Litomosoides anguyai n. sp., a parasite of the abdominal cavity of Oxymycterus misionalis, belongs to the sigmodontis group and is closely related to L. legerae and L. oxymycteri. The new species is differentiated by the salient amphids, an asymmetrical annular thickening of the buccal capsule, by the arrangement of the head and tail papillae, and the shape and size of the microfilaria. PMID- 12075132 TI - A discussion on the Heteracanthocephalidae Petrochenko, 1956 (Acanthocephala: Palaeacanthocephala). AB - Several heteracanthocephalid specimens were recovered from the flatfish Rhombosolea leporina (Gunther), a host of Heteracanthocephalus peltorhamphi (Baylis, 1944) Petrochenko, 1956 from New Zealand. Unlike H. peltorhamphi, these new specimens have trunk spines. Measurements and proboscis armament of the new specimens are consistent with the worms being Aspersentis minor Edmonds & Smales, 1992 originally described from the Australian flounder Rhombosolea tapirina Gunther. A review of the family Heteracanthocephalidae Petrochenko, 1956 was undertaken to assess the validity of its four genera and eight species. The validity of Aspersentis megarhynchus (Linstow, 1892) Golvan, 1960 (syn. Echinorhynchus megarhynchus Linstow, 1892) is questioned. E. megarhynchus is not considered to be an heteracanthocephalid and is relegated to a species inquirenda. A. megarhynchus (Linstow, 1892) of Golvan (1960) nec E. megarhynchus Linstow, 1892 is considered a synonym of A. austrinus Van Cleave, 1929. The monotypic genus Heteracanthocephalus Petrochenko, 1956 is proposed as a synonym of Aspersentis Van Cleave, 1929 because there appear to be insufficient morphological differences between them. Aspersentis peltorhamphi n. comb. is proposed for Heteracanthocephalus peltorhamphi. The monotypic genus Sachalinorhynchus Krotov & Petrochenko in Petrochenko, 1956 is considered valid, but the other heteracanthocephalid genus, Bullockrhynchus Chandra, Rao & Shyamasundari, 1985, also monotypic, is not. B. indicus Chandra, Rao & Shyamasundari, 1985 possesses more features resembling rhadinorhynchids than heteracanthocephalids but only females are known, and therefore the genus and species cannot be placed. There are currently four valid species of Aspersentis and one of Sachalinorhynchus. PMID- 12075133 TI - Respiration, copper availability and SOD activity in P. anserina strains with different lifespan. AB - P. anserina mutants with impairments in complex IV (COX) of the respiratory chain are characterized by an increase in lifespan. Examples are the nuclear grisea mutant with a moderate lifespan extension (60%) and the immortal extranuclear ex1 mutant. Here we report data demonstrating that in mutant ex1 the level of the alternative oxidase (PaAOX) is significantly higher than in mutant grisea. PaAOX levels appear to be reversely dependent on COX activity. The activity profile of superoxide dismutases in the ex1 mutant resembles the profile in senescent wild type cultures with a high cytoplasmic copper/zinc superoxide dismutase (PaSOD1) and a low mitochondrial manganese superoxide dismutase (PaSOD2) activity. In the grisea mutant, PaSOD1 activity is only detectable in cultures grown in copper supplemented medium. The two copper-regulated genes PaCtr3 (coding for a high affinity copper transporter) and PaSod2 are not expressed in the two mutants grown in standard medium. The repression of these genes as well as the activity of PaSOD1 is dependent on the availability of cellular copper, which appears to be high in COX-deficient strains such as mutant ex1 and in the senescent wild type strain. In the wild-type, changes in the cellular localization of copper and in the delivery of this metal to different proteins appear to occur during senescence. Collectively, the data explain the characteristic lifespan of the investigated strains as the result of differences in energy transduction and in the machinery protecting against oxidative stress. PMID- 12075134 TI - Temperature-induced changes in anaerobic capacity of interventricular septum in trained young and middle-aged rats. AB - The interventricular site has been proposed as one of the sites for the myocyte cell death with aging and thereby enhancing the possibility of ventricular dysfunction with aging. Variations in lactate dehydrogenase (LDH), a key glycolytic regulatory enzyme along with its isomers were used as indices to evaluate the anaerobic capacity of the interventricular septum (IVS) in 2-month old (young) and 12-month-old (middle-aged) rats trained to swim at 25 degrees C (cold, C) and 35 degrees C (thermo neutral, N) temperatures. Blood lactate (La) and LDH along with its isoenzymes in the IVS were assessed and compared with untrained and sedentary rats. Increased septal and heart weight was indicative of age-related hypertrophy accompanied by increased septal thickness. Elevated total LDH activity was evident in both the trained groups in the young. The middle-aged animals exhibited lesser variations in LDH and, H and M subunits. The data suggest an efficient adaptability of the IVS of young rats to physical training in contrast to a poor response by the IVS of middle-aged animals. PMID- 12075135 TI - Alterations in oxidative stress scavenger system in aging rat brain and lymphocytes. AB - There is a large body of evidence indicating an age-related increase in the rate of mitochondrial O2- and H2O2 generation and huge amounts of oxidative damage leading to several neurodegenerative disorders, perhaps due to an imbalance between free radical generation and anti-oxidant defense system. The aim of the present study was to elucidate the effect of aging on free radical scavenger system profile in rat brain and lymphocytes. The enzyme activities of gamma-GCS, GR, GPx, gamma-GTP, GST, catalase, and SOD as well as GSH content were assayed from discrete brain areas viz., CH, CB, BS and DC along with lymphocytes from four different age group rats, namely, 1-month-old young rats, 3-4-month-old young adults, 12-month-old adults and 24-month-old aged rats. Significant decline was observed in all the enzyme activities in 12- and 24-month-old rats as compared to 3-4-month-old young adult rats and also, 1-month-old rats showed lower levels of enzyme activities as compared to 3-4-month-old rats. The maximum scavenger system activity was found in the young adult rats (3-4 months) as compared to the remaining age groups. Lymphocytes and brain showed a parallel pattern of age-related alterations in the free radical scavenger system components. The analysis of such alterations is important in ultimately determining the basis of neuronal dysfunction associated with aging and also defining the nature of these changes may help to develop therapeutic means to cure not only elderly but also individuals suffering from certain organic or psychiatric disorders. PMID- 12075136 TI - A model of accelerated aging induced by 5-bromodeoxyuridine. AB - We consider a scheme of aging with two possible mechanisms of senescence processes: aging with apoptosis and necrosis for differentiated cells and a multistage process of malignant transformation. Our model describes the multistage phenomena of aging and carcinogenesis by a set of stochastic equations using multivariate and diffusion processes. This model fits experimental data on the acceleration of aging processes from postnatal exposure to 5 bromodeoxyuridine (BrdU), and the induction of genome instability. The methods of stochastic modeling and computer simulation of the processes of aging and carcinogenesis have been used for the verification of the biological suggestions. PMID- 12075137 TI - Molecular chaperones and the process of cellular immortalization in vitro. AB - It is suggested that the chaperone-activity of the SV40 T protein is responsible for the first step in the process of cellular immortalization in vitro: the extension of life span of most transfected cells. Further that an additional up regulation of selected molecular chaperones is causal in the second step of the process: the rare acquisition of an unlimited cell division potential. Possibly the molecular chaperones are evolution facilitators, enabling the immortalization of primary cells in vitro, as well as the evolution of longevity in species. PMID- 12075138 TI - Lifespan does not measure ageing. AB - The Gompertz equation, which describes the increase of mortality over time, is often used to measure the rate of the ageing process. However, a recent article argued that it is incorrect to use the function for this purpose and that its use should therefore be discouraged (Driver 2001). I believe that this conclusion is based on misunderstandings and that the Gompertz function (if used appropriately) is actually well suited to measure the ageing rate. Here I will solve the 'paradox' posed by the author and argue that it is actually lifespan that is inadequate to measure ageing and not the Gompertz function. PMID- 12075141 TI - Adult learning, objectivity and other self-evident untruths. PMID- 12075142 TI - Assessing the comparability of standardized patient and physician evaluations of clinical skills. AB - Accumulating evidence to defend decisions based on scores from evaluations is an ongoing process. The purpose of this investigation was to gather additional data to support the validity of inferences made from scores on the Educational Commission for Foreign Medical Graduates' Clinical Skills Assessment (CSA). This was accomplished by contrasting CSA candidate scores, and pass/fail decisions, with those obtained from the American Board of Internal Medicine's Mini-CEX (Clinical Evaluation Exercise). Data gathering performance based on the number of unweighted history taking and physical examination checklist items adequately predicted the global ratings provided by physician observers. CSA ratings of doctor-patient communication skills correlated with mini-CEX ratings of like constructs, indicating that physician observers, using mini-CEX rating scales, are able to make realistic assessments of interpersonal skills. These results provide evidence of the convergent validity of CSA scores. PMID- 12075143 TI - Medical students' ratings of faculty teaching in a multi-instructor setting: an examination of monotonic response patterns. AB - Realizing that the psychometric properties of a measure may be highly variable is especially relevant in a multi-instructor context, since an implicit assumption is that student ratings are equally reliable and valid for all faculty ratees. As a possible indicator of nonattending (i.e. invalid) responses, the authors examined the effects of monotonic response patterns on the reliabilities of students' ratings of faculty teaching - including how an alternative presentation format may reduce the prevalence of this behavior. Second-year medical and dental students (n = 130) enrolled in a required basic science course during the 1998-99 academic year were randomly assigned to one of two groups - each of which evaluated the teaching of 6 different faculty across 6 distinct dimensions (i.e. overall quality, organization, preparation, stimulation, respectfulness, and helpfulness). Using a 'split ballot' design, two versions of the conceptually equivalent faculty evaluation form were distributed at random to students in each group. Form A contained the 'traditional' items-within-faculty format, while Form B listed faculty-within-item.The number of monotonic forms (i.e. the identical rating of all 6 items) varied measurably across faculty ratees, as did the respective effects on scale reliabilities. Alpha was especially inflated where a sizeable proportion of monotonic patterns were located on response categories that were either very high (> +1.28 z(m) deviations) or very low (< -1.28 z(m) deviations) compared to the group mean. Lastly, the prevalence of monotonic response patterns was significantly (p = < or = 0.01) less when a faculty-within item format is used (Form B). These findings suggest that monotonic response patterns differentially impact the reliabilities and, hence, the validity of students' ratings of individual faculty in a multi-instructor context. PMID- 12075144 TI - Learning to use minimal access surgical instruments and 2-dimensional remote visual feedback: how difficult is the task for novices? AB - Performing minimal access surgery requires the use of 2-dimensional information to produce 3-dimensional movements, as well as precise motor control for manipulating laparoscopic tools. The added visuomotor demands of this task make it more demanding and complex than traditional open surgery. The purpose of this study was to determine the relative task difficulty of performing laparoscopic tool movements with normal vision or 'laparoscopic vision' provided by a remote 2 D monitor. A second purpose of this study was to evaluate whether movement performance changes are induced by practice with normal vision (NV) and laparoscopic vision (LV). The study was also designed to determine whether order of visual condition (NV or LV) practice impacts the rate of performance acquisition when transferred to the opposing visual condition. Eleven individuals participated in this study. All subjects performed a bean grasping and a suturing task in two visual conditions: normal vision and laparoscopic vision. Results revealed that laparoscopic tools themselves do not appear to be problematic in performing minimal access surgery. Furthermore, performance ability in normal vision does not positively transfer to performance when switched to a laparoscopic vision condition. The 2-dimensional video does appear to be problematic for skill acquisition, as performance levels decreased as complexity of the task increased. PMID- 12075145 TI - Optimising the reproducibility of a performance-based assessment test in midwifery education. AB - Despite problems concerning generalisability of test results - largely due to limited sampling of the task domain - and questions about test efficiency, the popularity of OSCEs in medical education has motivated the use of similar assessment methods in other health care domains. Purpose of this study was to investigate reproducibility of scores on an OSCE-based test in the relatively narrow domain of midwifery, as compared to the broad medical domain. The influence of global rating scales and adoption of a mastery-oriented test perspective on reproducibility of test scores was investigated in order to explore possibilities to increase test efficiency.A 3-hr, 6-station performance based test was administered to third and fourth year students at Kerkrade Midwifery School (the Netherlands). Students' performance was recorded using a station specific checklist and a global rating scale. For the 3-hr OSCE the generalisability coefficient based on checklist scores is 0.48. Based on global ratings alone, the generalisability coefficient is 0.61. Adjusted dependability indices are 0.85 based on checklist scores and 0.95 for global ratings respectively. Results suggest that even for small domains the problem of case specificity remains a major impediment to high stakes performance testing. Findings furthermore suggest that use of global rating scales and professional expert judgements in professional competence evaluation is to be preferred to task specific checklists for reasons of reproducibility and efficiency in test development and administration. Adoption of a mastery oriented test perspective may reduce testing time requirements even further. PMID- 12075146 TI - Systematic reviews: what are they and why should we care? PMID- 12075147 TI - Andragogy and medical education: are medical students internally motivated to learn? AB - Andragogy - the study of adult education - has been endorsed by many medical educators throughout North America. There remains, however, considerable controversy as to the validity and utility of adult education principles as espoused by the field's founder, Malcolm Knowles. Whatever the utility of andragogic doctrine in general education settings, there is reason to doubt its wholesale applicability to the training of medical professionals. Malcolm Knowles' last tenet of andragogy holds that adult learners are more motivated by internal than by external factors. The validity of this hypothesis in medical education is examined, and it is demonstrated that medical students' internal and external motivation are context-dependent, not easily distinguishable, and interrelate with one another in complex ways. Furthermore, the psychological motivation for medical student learning is determined by a variety of factors that range from internal to external, unconscious to conscious, and individual to societal. The andragogic hypothesis of increased internal motivation to learn on the part of adults in general, and medical trainees in particular, is rejected as simplistic, misleading, and counterproductive to developing a greater understanding of the forces that drive medical students to learn. PMID- 12075148 TI - Molecular variation of Bothriocephalus acheilognathi Yamaguti, 1934 (Cestoda: Pseudophyllidea) in different fish host species based on ITS rDNA sequences. AB - The molecular variation in Bothriocephalus acheilognathi Yamaguti, 1934 from 11 species of freshwater fish collected in Australia, China, the Czech Republic, England and Hawaii was investigated by determining the nucleotide sequences of the internal transcribed spacer region. The length of the first and second internal transcribed spacer sequences of multiple individuals ranged from 553 to 571 bp and 553 to 615 bp, and the G + C content from 53.1 to 53.5%. The percentage sequence divergence varied between 0 and 0.9% in the ITS1 and 0 and 6.6% in the ITS2, respectively, indicating the occurrence of intraspecific variation. It is demonstrated that the fragment length variation resulted primarily from microsatellite polymorphisms present in the ITS region, especially in the ITS2 region. Phylogenetic analyses revealed that B. acheilognathi examined in this study consisted of three closely related genotypes with certain degrees of host-specificity, and the genotype representing isolates from Cyprinus carpio L. was the most common and diverse form within the species B. acheilognathi. PMID- 12075149 TI - A new species of Eudusbabekia Jameson, 1971 (Acari: Myobiidae) parasitising Rhinophylla pumilio Peters (Chiroptera: Phyllostomidae) from Ecuador. AB - The mite Eudusbabekia rhinophylla n. sp. is described from the bat Rhinophylla pumilio Peters in Ecuador. It is considered to be a member of the subgenus Eudusbabekia, differing from other representatives of this group by the lanceolate shape of the coxal setae. PMID- 12075150 TI - Aptorchis aequalis Nicoll, 1914 (Digenea: Plagiorchiidae) is a senior synonym of Dingularis anfracticirrus Jue Sue & Platt, 1999 (Digenea: Plagiorchiidae). AB - Examination of the type-specimen of Aptorchis aequalis Nicoll, 1914 (Digenea: Plagiorchiidae) revealed anatomical features not included in the original description; most notably the intestinal caeca extend to near the posterior end of the worm instead of terminating just posterior to the ventral sucker. A comparison of the type of A. aequalis with specimens identified as Dingularis anfracticirrus Jue Sue & Platt, 1999, from the author's collection yielded no differences that would justify the recognition of two genera. Therefore, Dingularis Jue Sue & Platt, 1999, is considered a junior subjective synonym of Aptorchis Nicoll, 1914. Principal components analysis of specimens of Aptorchis collected from three species of freshwater turtles at various localities in Queensland and New South Wales, Australia revealed no differences between these specimens and the type-specimen of Dingularis anfracticirrus. Therefore, D. anfracticirrus is considered a junior subjective synonym of A. aequalis. Aptorchis Nicoll, 1914, includes the following species: A. aequalis (= Dingularis anfracticirrus new synonymy); A. pearsoni (Jue Sue & Platt, 1999) n. comb.; and A. megapharynx (Jue Sue & Platt, 1999) n. comb. PMID- 12075151 TI - Redescription of Raphidascaris gigi Fujita, 1928 (Nematoda: Anisakidae), a parasite of freshwater fishes in Japan. AB - A redescription is given of Raphidascaris gigi Fujita, 1928 based on newly collected specimens from the intestine of masu salmon Oncorhynchus masou (Brevoort) in the Kinu River in Japan. The species is characterised by the absence of interlabia and lateral alae, by the presence of lips without lateral membranous flanges and by numerous pre-anal and postanal papillae in the male. Although the general morphology of this species resembles that of Ichthyascaris spp. as defined by Bruce (1990), who transferred R. biwakoensis (= R. gigi) to this genus, the absence of the alae uniting close the ventrolateral lips shows clearly that it belongs to Raphidascaris Railliet & Henry, 1915. Ichthyascaris Wu, 1949 is here considered a subgenus of Raphidascaris, and two species described in the former genus are transferred to the latter as R. gymnocraniae (Bruce, 1990) n. comb. and R. sillagoides (Bruce, 1990) n. comb. The finding of R. gigi in O. masou represents a new host record. PMID- 12075152 TI - Isthmiophora Luhe, 1909 and Euparyphium Dietz, 1909 (Digenea: Echinostomatidae) re-defined, with comments on their nominal species. AB - The validity of Isthmiophora Luhe, 1909 in relation to Euparyphium Dietz, 1909 is discussed and confirmed. Isthmiophora melis Schrank, 1788) [the type-species] and I. inermis (Fuhrmann, 1904) n. comb. are redescribed, and diagnoses are given for both genera, along with lists of their presently-accepted constituent species which are commented upon where necessary. A similar list of species previously allocated to these genera is also presented with comments on their current status. A key to the species of Isthmiophora is included. New combinations for species previously attributed to Euparyphium are: Isthmiophora inermis (Fuhrmann, 1904) n. comb., I. beaveri (Yamaguti, 1958) n. comb., I. lukjanovi (Chertkova, 1971) n. comb., I. citellicola (Kadenatsii in Skrjabin & Bashkirova, 1956) n. comb., I. hortensis (Asada, 1926) n. comb., Echinostoma pindchi (Khan & Chishti, 1985) n. comb., Echinoparyphium tripathii (Gupta & Gupta, 1982) n. comb., E. hirundonis (Fischthal & Kuntz, 1976) n. comb., and Hypoderaeum longitestis (Verma, 1936) n. comb. Species attributed to Euparyphium which are here considered species inquirendae are: E. lobata Farooq & Yousuf, 1986 sp. inq., E. ochoterenai Cerecero, 1943 sp. inq., E. sobolevi Ryzhikov, 1965 sp. inq., and E. taiwanense Fischthal & Kuntz, 1976 sp. inq. PMID- 12075153 TI - Elaeolenchus parthenonema n. g., n. sp. (Nematoda: Sphaerularioidea: Anandranematidae n. fam.) parasitic in the palm-pollinating weevil Elaeidobius kamerunicus Faust, with a phylogenetic synopsis of the Sphaerularioidea Lubbock, 1861. AB - A new nematode, Elaeolenchus parthenonema n. g., n. sp., is described from the palm-pollinating weevil Elaeidobius kamerunicus Faust. The new genus is placed in the Anandranematidae n. fam., which, together with the genus Anandranema Poinar et al., 1993, is characterised by nematodes having only a single autotokous generation in the insect host. This is the first report of a member of this superfamily reproducing only parthenogenetically. The development of E. parthenonema and its effect on the weevil host is discussed, along with a phylogenetic synopsis of the families of the Sphaerularioidea Lubbock 1861. The Beddingiidae n. fam. is proposed for Beddingia Blinova & Korenchenko, 1986, comprising the original Deladenus parasites of Hymenoptera that possess both free living and parasitic amphimictic generations in their life-cycles. This family is considered to have the most primitive type of development in the superfamily. PMID- 12075154 TI - Synaptobothrioides sardinellae n. g., n. sp. (Digenea: Hemiuridae) from the blacktip sardinella Sardinella melanura (Clupeidae) off Fiji, and records of two other hemiuroids in Fijian waters. AB - Synaptobothrioides sardinellae n. g., n. sp. is described from Sardinella melanura off Fiji. The new genus differs from other plerurine hemiurid genera in having entire or angular, rather than lobed to digitiform, vitelline masses. It appears closest to Synaptobothrium, but differs in the lack of a presomatic pit and the presence of tegumental plications. Lecithochirium kawakawa from Euthynnus affinis and Thulinia microrchis from Siganus spinus are reported from off Fiji. PMID- 12075155 TI - [Breast cancer screening]. PMID- 12075156 TI - [Breast cancer screening]. AB - Since 1989, several breast cancer screening mammography programs have been established. The purpose of this chapter is to provide the history and the main results of the previous programs. Furthermore, special attention is given to the new rules established in 2001 with emphasis of the general principles as well as the ethical principles of breast screening. PMID- 12075157 TI - [How to read a screening mammography (mammotest)]. AB - The purpose of this chapter is to give to the radiologist some practical information in order to be able to read the mammotests, as they are performed in mass screening for breast cancer in France. The examples shown must help to detect as many small cancers as possible, without inflating the false positive rate. Screening mammotests can be read one by one (first reading), or by large series (second or third reading). To improve both the sensitivity and the specificity, reading technique should be strict, with correct fitting (viewboxes, ambiant light). For each case, a technical evaluation is performed (sharpness, contrast, breast projection), followed by the detection phase (asymmetry, abnormal density or calcifications or distorsion). Then, the image is analyzed (eliminating the typical benign conditions), and finally encoded to formalize the result: normal/suspicious for breast cancer. A good working method and a perfect knowledge of normal and pathologic appearances, allow us to reach the recommended rates for detected cancers and positive tests. PMID- 12075159 TI - [Using BI-RADS in French]. AB - BI-RADStrade mark is the referral lexicon in the United States for the classification of terms which are used for reporting breast imaging. The purpose of this chapter is to present and summarize the French version. PMID- 12075160 TI - [Screening and imaging guided biopsies of the breast]. AB - In cases of subclinical mammographic abnormalities, being able to determine after screening, the indications regarding the type of imaging guidance and the type of biopsy equipment. In presence of microcalcifications, stereotactic mammography, either screen-film or digital, is the modality of choice. As fine needle aspiration cytology is insufficient, it is necessary to obtain tissue specimens with at least 14 Gauge Tru-cut needle, triggered by automatic guns, or coaxial needles of 11 G or 8 G, adaptable on a vacuum aspiration system. In case of mammographic opacities or masses, the same type of sampling can be used under US guidance as long as there is a good sonographic contrast of the lesion. Fine needle puncture, well accepted, keeps all its interest for cystic lesions and for solid ones, because of the value of its multidirectionnal sampling that well trained cytopathologists can make very informative. However, in case of insufficient or discordant results, US guided microbiopsies or macrobiopsies, using newer guns, allow to obtain a histologic diagnosis equivalent to surgical biopsy. If fine needle US guided puncture can be performed in first intention, straight after ultrasonographic examination, percutaneous micro- or macrobiopsies (using US or stereotactic guidance) must be undertaken in second intention after having explained to the patient this ambulatory procedure, the eventual risks, and by insisting on the results that can be expected for a precise diagnosis. PMID- 12075161 TI - [Clinical case. Screening and imaging guided biopsies of the breast]. PMID- 12075162 TI - [Radiologic-pathologic correlations in breast diseases]. AB - The objective of this article is to explain radiologic patterns of benign and malignant breast lesions (masses, microcalcifications) based on histological correlations. The stromal fibrous reaction associated to infiltrating carcinomas is responsible of focal increased density, and architectural distorsion, ultrasound acoustic shadowing; abnormal neoangiogenesis can be detected by Doppler, CT or MR imaging. Invasive carcinomas without spiculated margins are poorly differentiated tumors. Mammographic patterns of microcalcifications depend on their physiopathological process (necrosis, secretion), and the shape of clusters (round, triangular) typifies their anatomical site of origin (lobular, ductal). Less frequent lesions (invasive lobular, mucinous, and medullary carcinomas, radial scar) will be also explained based on radiopathological correlations. Knowledge of radiopathological correlations in breast diseases helps the radiologists to analyze and characterize breast lesions. PMID- 12075163 TI - [Clinical case. Radiologic-pathologic correlations in breast diseases]. PMID- 12075164 TI - [Magnetic Resonance Imaging and PET of the breast]. AB - Breast MR imaging and PET allow physiopathological tissular approach of breast tumors. Breast MR imaging evaluates tumoral angiogenesis whereas PET analyzes metabolism of tumoral cells. Since 1985, breast MR imaging has been extensively studied. All these series agreed in the high sensitivity of breast MR imaging for detecting invasive breast cancers whereas the sensitivity varies from 60 to 85% in detecting ductal carcinoma in situ. However, its reported specificity varies between 60 and 97%, mainly due to some entities such as fibroadenomas and fibrocystic disease. Clinical indications are the diagnosis of local recurrences, the evaluation of pre-operative chemotherapy and breast prosthesis. PET of the breast is currently in clinical research phase. Early results are promising especially in the evaluation of pre-operative chemotherapy and diagnosis of isolated axillary nodes. PMID- 12075165 TI - [Clinical case 1. Magnetic Resonance Imaging and PET of the breast]. PMID- 12075166 TI - [Clinical case 2. Magnetic Resonance Imaging and PET of the breast]. PMID- 12075167 TI - [Digital mammography and computer assisted diagnosis]. AB - Until recently, the film has remained the only medium of information in mammography. The film is used to record, to exploit, to store and to transmit the image. Computerization of images allows to dissociate and to optimize these different functions. Among classical factors of image quality (spatial resolution, contrast, noise), new factors should be added like the detective quantum efficiency and the conversion factor. Radioluminescent screens, then digital sensors for breast stereotactic imaging have been marketed. Manufacturers are now testing full field digital mammographs. Digital imaging allows many applications (computed-aided diagnostic, 3D imaging.) and permits the easy transfer of images for diagnosis and teaching. Three parts are presented in this chapter. The first one describes the different imaging modalities and gives a reminder of the different elements related to image quality. The second one is related to the practical aspects of full field mammography, the reading of mammograms on a review station, ergonomy in full field mammography and to possible changes for screening mammography. The third part is devoted to computed aided diagnosis and its possible application in screening. PMID- 12075168 TI - [Clinical case. Digital mammography and computer assisted diagnosis]. PMID- 12075171 TI - Ubiquitin proteasome inhibition and cancer therapy. PMID- 12075172 TI - Signal transduction by reactive oxygen species: alternative paradigms for signaling specificity. PMID- 12075173 TI - Functional characteristics of the pylorus in patients undergoing pylorus- preserving gastrectomy for early gastric cancer. AB - BACKGROUND: This study investigates the functional characteristics of the pylorus in patients undergoing pylorus-preserving gastrectomy (PPG) for early gastric cancer. METHODS: In study 1, postprandial symptoms and gastric emptying were compared between 2 groups of 12 patients with early gastric cancer more than 1 year after either PPG (PPG group) or distal gastrectomy (DG) (DG group). Gastric emptying was evaluated with the dual isotope technique for liquids and solids separately. In study 2, pyloric motility was evaluated with a sleeve/sidehole manometric assembly in 7 patients undergoing PPG, which was compared with that in the preoperative patients. RESULTS: In study 1, the overall modified Visick score of postprandial symptoms and the Sigstad dumping score were significantly lower in the PPG group compared with the DG group (P <.05). Early accelerated gastric emptying was observed in both groups for liquids, but only in the DG group for solids. In study 2, isolated pyloric pressure waves induced by intraduodenal lipid infusion and phase III-like activity induced by intravenous erythromycin infusion were preserved after PPG. CONCLUSIONS: The quantitative analysis of postoperative symptoms revealed that PPG patients were associated with better clinical conditions than DG patients. The clinical benefits of PPG are considered to be based on the function of the preserved pylorus. PMID- 12075174 TI - Lap-band failures: conversion to gastric bypass and their preliminary outcomes. AB - BACKGROUND: The LAP-BAND is designed to be an adjustable laparoscopically placed gastric restriction device for the treatment of severe obesity. The purpose of this study was to assess the outcome in patients who had failed to effectively lose weight with this device and were converted to a gastric bypass. METHODS: A retrospective chart review was performed of all LAP-BANDS placed in patients at our institution from March 1996 to June 1998. RESULTS: 36 LAP-BANDS were placed. To date, 18 of 36 (50%) have been removed. Fourteen of 18 were simultaneously converted to a gastric bypass. Indications for conversion included: failed weight loss (5), failed weight loss with esophageal dilatation (5), failed weight loss with leaking band (2), and symptomatic esophageal dilatation (1). Median time to conversion after LAP-BAND placement was 38.2 months. Median follow-up after conversion to gastric bypass was 8.3 months. Nineteen percent excess weight loss occurred after LAP-BAND placement. Forty-three percent excess weight loss occurred after conversion to gastric bypass (P =.025). CONCLUSIONS: In our experience, the LAP-BAND is associated with a high frequency of inadequate weight loss. Conversion to gastric bypass in this subset of patients is technically challenging but results in superior weight loss in a shorter time period. PMID- 12075175 TI - Lymph node micrometastasis and lymphatic mapping determined by reverse transcriptase-polymerase chain reaction in pN0 gastric carcinoma. AB - BACKGROUND: The purposes of this study were to examine lymph node micrometastasis (LMM) by reverse transcriptase-polymerase chain reaction (RT-PCR) method, and clarify the initial nodes involved by metastatic disease according to tumor location. METHODS: We examined 312 lymph nodes obtained from 50 patients with node-negative gastric carcinoma. RT-PCR and immunohistochemistry were performed. The clinical characteristics of LMM were investigated, and the map of LMM was made according to tumor location. RESULTS: The number of patients and LMM detected by RT-PCR was 14 and 17 and by immunohistochemistry was 7 and 8, respectively. RT-PCR was a more sensitive method than immunohistochemistry. LMM by RT-PCR correlated with depth of tumor invasion and lymphatic invasion. Regarding pT1 tumor, 9 patients with LMM had tumors that were of the macroscopically depressed type and 2 cm or more in diameter. According to the lymphatic map, right pericardial lymph nodes and lymph nodes along the lesser curvature were the initial nodes involved in the upper third of the stomach. Right pericardial lymph nodes, lymph nodes along the lesser curvature, and infrapyloric nodes were more important initial metastatic sites in the middle third of the stomach, and lymph nodes along the lesser curvature and infrapyloric nodes in the lower third. CONCLUSIONS: We demonstrated the relationship between LMM and clinicopathologic factors, especially in pT1 tumor. The mapping of LMM may prove useful for selecting the optimal treatment. PMID- 12075176 TI - The role of operations for distantly metastatic well-differentiated thyroid carcinoma. AB - BACKGROUND: The role of operations for distantly metastatic well-differentiated thyroid carcinoma (DTC) is poorly defined. We review the indications for operation for metastatic DTC. METHODS: This study consists of 260 patients treated between 1941 and 2000 for metastatic DTC, of which 59 (23%) underwent operations. Median follow-up was 7 years (range, 1 to 49 years). Metastases were identified clinically in 157 (60%) and radiologically in 103 (40%) patients. The disease-specific survival was estimated with the Kaplan-Meier method. RESULTS: Twenty-four patients (9%) were disease-free with resection. Palliative resection was indicated for painful bone metastasis, pathologic fracture, or symptomatic spinal cord involvement (35/260, 14%). Patients who could undergo complete metastasectomy survived longer than those having incomplete/palliative resection or nonoperative treatment for metastatic DTC (5-year disease-specific survival, 78% vs 43% vs 46%, P =.03). CONCLUSIONS: Solitary distant metastasis of DTC amenable to complete resection is infrequent. Complete metastasectomy may be associated with improved survival for localized distant disease. Palliative resection is indicated to improve quality of life for symptomatic distant metastasis. PMID- 12075177 TI - Intraluminal gene transfer of endothelial cell-nitric oxide synthase suppresses intimal hyperplasia of vein grafts in cholesterol-fed rabbit: a limited biological effect as a result of the loss of medial smooth muscle cells. AB - BACKGROUND: The intimal hyperplasia of vein grafts is a major cause of late graft failure and is more pronounced under hyperlipidemia. We previously reported that endothelial cell (ec)-type nitric oxide synthase (NOS) gene transfer inhibited graft intimal hyperplasia under poor runoff conditions. However, little information is available on either ecNOS gene transfer or intimal thickening under hypercholesterolemia. METHODS: Using the hemagglutinating virus of Japan liposomes, bovine ecNOS complentary DNA (5000 hemagglutinating activity units/mL) was transfected intraluminally to the right jugular vein, and these veins were then implanted as reversed vein grafts in an end-to-side fashion to the ipsilateral carotid artery. RESULTS: The cyclic guanosine 3',5'-monophosphate content of the ecNOS vein significantly increased in the grafts at 4 days after gene transfer, but the levels were only 25% greater than those found in the untreated veins. An immunohistochemical analysis at the same time suggested a large loss of medial smooth muscle cells that might have led to a reduction in the exogenous gene expression. The neointima of the ecNOS grafts was significantly reduced 4 weeks after implantation (P <.05), but the effect of ecNOS was limited to about a 30% inhibition. This reduction was associated with a reduced population of proliferating cells and decreased macrophage accumulation in the graft wall. CONCLUSIONS: These results demonstrated that the ecNOS gene transfer suppressed intimal hyperplasia of the vein grafts under hyperlipidemic conditions. However, this effect may be limited because of the smooth muscle cell loss related to the use of an intraluminal delivery methods. These data lead to speculation that the outcome of ecNOS gene transfer could be improved using different methods of gene delivery. PMID- 12075178 TI - Attenuation of renal ischemia-reperfusion injury by FR167653 in dogs. AB - BACKGROUND: Inflammatory cytokines are known to contribute to ischemia reperfusion injury. We investigated the effect of FR167653 (FR), a suppressor of interleukin-1beta and tumor necrosis factor-alpha, on ischemia-reperfusion injury of the kidney in dogs. METHODS: The left kidney was subjected to ischemia for 60 minutes followed by removal of the right kidney. A control group (n = 10) and an FR group (n = 8) were evaluated for tissue blood flow; resistive index, pulsatility index, arterial oxygen pressure, serum creatinine, blood urea nitrogen, aspartate transaminase, and alanine transaminase levels; interleukin 1beta messenger RNA expression in the peripheral blood; apoptotic index; and histopathology. RESULTS: The FR group showed lower creatinine, serum urea nitrogen, aspartate transaminase, and alanine transaminase levels (P <.038 each) and lower interleukin-1beta mRNA expression and apoptotic index (P <.041 each) than did the control group. Arterial oxygen pressure during the 120 minutes after reperfusion in the FR group decreased but recovered quickly (P =.024). Renal tissue damage in the FR group was less than that in the control group (P =.036). CONCLUSIONS: FR ameliorates ischemia-reperfusion injury of the kidney potentially by reduced production of inflammatory cytokines that may contribute to damage to the ischemic kidney and the distant organs. PMID- 12075179 TI - Lipstick and pearls. PMID- 12075181 TI - Perspectives of a woman resident. PMID- 12075182 TI - Success, women, and academic surgery. PMID- 12075183 TI - Leaders in American surgery: where are the women? PMID- 12075184 TI - Double common bile duct: a case report and a review of the Japanese literature. AB - BACKGROUND: A double common bile duct (DCBD) is a rare congenital anomaly. We report the case of a 60-year-old Japanese female, whose common bile duct divided into 2 channels and both channels opened individually into the second portion of the duodenum. This is the fourth reported case of DCBD with a choledochal cyst and pancreaticobiliary maljunction (PBM). METHODS: A review of the literature revealed that DCBD is more frequently diagnosed in Oriental people. We reviewed 47 cases of DCBD reported in the Japanese literature. RESULTS: Among these, cholelithiasis was found in 27.7%, a choledochal cyst in 10.6%, PBM in 29.8%, and cancers in 25.5%. Cancer and PBM were the 2 most serious concomitant conditions. The incidence and type of complicating cancer and PBM varied according to the site of the opening of the accessory common bile duct (ACBD). Concomitant gastric cancer was frequently noted when the ACBD opened into the stomach, whereas cancer of the biliary system was common when the ACBD opened into the second portion of the duodenum or the pancreatic duct. PBM was observed only in those patients in whom the ACBD opened into the second portion of the duodenum or the pancreatic duct. Therefore the treatment and prognosis of DCBD is influenced by the site of opening of the ACBD. CONCLUSIONS: In DCBD, the opening site of the ACBD was considered to have close implications for the type of concomitant cancer and concomitant PBM that would appear. PMID- 12075185 TI - A case of adult-to-adult living donor liver transplantation using right and left lateral lobe grafts from 2 donors. PMID- 12075186 TI - Multiple gastric perforations in an immunodepressed child. PMID- 12075187 TI - Giant colonic lipoma as lead point for intermittent colo-colonic intussusception. PMID- 12075188 TI - Patent omphalomesenteric duct. PMID- 12075189 TI - End-to-end anastomosis of pancreas. PMID- 12075190 TI - Operative techniques in inguinal hernia repair associated with patients returning to work. PMID- 12075192 TI - Acute hypersplenism with splenomegaly after portal vein embolization. PMID- 12075193 TI - Gallbladder carcinoma. PMID- 12075195 TI - Imaging of the jaws for routine implant placement. PMID- 12075196 TI - Smokeless tobacco use and cancer of the upper respiratory tract. AB - The most recent epidemiologic review of the cancer risks associated with smokeless tobacco use appeared in 1986, when 10 studies were available. This review describes 21 published studies, 20 of which are of the case-control type. We characterize each study according to the specific anatomic sites and according to the type of smokeless tobacco products for which it provides relative risks of cancer. The use of moist snuff and chewing tobacco imposes minimal risks for cancers of the oral cavity and other upper respiratory sites, with relative risks ranging from 0.6 to 1.7. The use of dry snuff imposes higher risks, ranging from 4 to 13, and the risks from smokeless tobacco, unspecified as to type, are intermediate, from 1.5 to 2.8. The strengths and limitations of the studies and implications for future research are discussed. PMID- 12075197 TI - Tender preauricular swelling. PMID- 12075198 TI - Alzheimer's disease and dental management. AB - Alzheimer's disease (AD) is a major disorder of old age and the most common cause of dementia. Dementia is loss of intelligence, memory, and cognitive functions. It is usually associated with aging, but there are many possible causes. Older adults are the most rapidly growing segment of the population. To maintain lifelong good oral health, the elderly need regular dental care. This review of AD summarizes the causes, epidemiology, diagnosis, and dental management of patients with AD. PMID- 12075199 TI - Salivary stone lithotripsy in the HIV patient. AB - OBJECTIVE: To investigate the use of shock-wave lithotripsy in the treatment of salivary gland disease in HIV-positive patients. STUDY DESIGN: Four patients infected with human immunodeficiency virus with ultrasonographically confirmed sialolithiasis (three male patients, mean age 33.5 years, range 19-41 years) were treated with extracorporeal electromagnetic shock-wave lithotripsy. RESULTS: All but one of the patients were successfully treated or experienced relief, with complete stone clearance demonstrated by ultrasonography 12 months after lithotripsy. CONCLUSION: Extracorporeal shock-wave lithotripsy is a safe, effective and minimally invasive technique for the nonsurgical treatment of HIV positive patients with sialolithiasis. PMID- 12075200 TI - Preoperative histologic assessment of head and neck lesions using cutting needle biopsy. AB - OBJECTIVE: This study assessed the clinical utility of cutting needle biopsy in which a newly developed Monopty biopsy instrument (MBI) (Monopty, Bard Urologic Division; Covington, Ga) was used in the preoperative assessment of head and neck lesions. STUDY DESIGN: Needle biopsies were performed with the MBI in 16 cases of head and neck lesions that included lesions in lymph nodes, salivary glands, palate, and soft tissue. RESULTS: High-quality histopathologic specimens were obtained without complications in all biopsies performed, and the diagnostic target tissue was obtained in 15 of 16 cases. Diagnoses made from MBI needle biopsy specimens were consistent with the final diagnoses made from subsequent surgical materials in 14 cases, and the accuracy rate was 88%. None of the samples demonstrated significant rush artifacts or obscuring blood, both of which are problems commonly associated with manual biopsy techniques. CONCLUSIONS: This technique offers a safe and effective means of obtaining adequate tissue for the histological assessment of head and neck lesions. PMID- 12075201 TI - Mandibular, sagittal split osteotomies fixed with biodegradable or titanium screws: a prospective, comparative study of postoperative stability. AB - OBJECTIVE: Comparison of skeletal stability following bilateral sagittal split osteotomy (BSSO) advancement of the mandible fixed with titanium or biodegradable bicortical screws. STUDY DESIGN: Forty consecutive patients underwent mandibular advancement by means of BSSO performed with a standardized technique. In 20 patients rigid fixation was achieved by means of titanium bicortical screws; the other 20 patients were fixed with biodegradable copolymer screws made of poly-L lactic acid (82%) and polyglycolic acid (18%). Lateral cephalograms were obtained 1 week preoperatively, 1 week postoperatively and after a minimum of 6 months postoperatively. Relevant skeletal points were traced and digitized to evaluate 2 dimensional skeletal change. Changes at each time point were analyzed and compared statistically. RESULTS: There was no statistically significant difference in long-term stability between the 2 groups. No clinical or radiographic evidence of wound healing problems were noted. CONCLUSION: Resorbable poly-L-lactic/polyglycolic acid copolymer bicortical screw fixation of a BSSO is a viable alternative to titanium screws for the fixation of advancement BSSO. PMID- 12075202 TI - Effect of maxillary distraction osteogenesis on velopharyngeal function: a pilot study. AB - OBJECTIVE: The purpose of this study was to examine preoperative and postoperative changes of velopharyngeal function in cleft patients who underwent maxillary distraction osteogenesis using the Rigid External Distraction System. STUDY DESIGN: Six cleft patients followed for a minimum of 12 months after maxillary distraction were examined. Plain and contrast lateral-cephalograms were obtained preoperatively and postoperatively, and speech evaluation was performed by the same authorized speech therapist at the same time points. RESULTS: The mean distraction amount at the anterior nasal spine was 11.7 mm (range, 7.4 mm - 15.0 mm). Both the nasopharyngeal depth and velar length increased after maxillary distraction, but the need ratio (nasopharyngeal depth/velar length) also increased after distraction. Although scores for velopharyngeal closure dropped a few points after maxillary distraction, the rating for hypernasality remained unchanged in all patients but the patient whose distraction amount was 15.0 mm. CONCLUSION: These results suggest that maxillary distraction of less than 15 mm may not markedly affect velopharyngeal function in cleft patients. PMID- 12075203 TI - Antithrombotic agents: implications in dentistry. AB - Thrombosis and the complicating emboli that can result are important causes of illness and death. Thrombosis is of greater overall clinical importance in terms of morbidity and mortality than all of the hemorrhagic disorders combined. Agents such as heparin, low-molecular weight heparin, warfarin, aspirin, ticlopidine, clopidogrel, and tirofiban are used to prevent venous or arterial thrombosis. Patients taking these antithrombotic agents may be at risk for excessive bleeding after invasive dental procedures. The current antithrombotic agents used in medicine are reviewed, and the dental management of patients taking these agents is discussed. PMID- 12075204 TI - The value of maximum jaw motion measurements for distinguishing between common temporomandibular disorder subgroups. AB - OBJECTIVE: The purpose of this study was to determine if mandibular motion measurements could be used to distinguish between common temporomandibular disorder (TMD) subgroups that were established on the basis of only clinical signs and symptoms. STUDY DESIGN: Patients were 41 consecutive TMD clinic patients (31 women and 10 men). These patients were divided into 6 typical TMD subgroups. The subgroups were patients with (1) arthromyalgia, (2) arthromyalgia with disk condyle incoordination, (3) disk condyle incoordination only, (4) osteoarthritis, (5) suspected disk displacement without reduction, or (6) other diagnoses. RESULTS: There were no subjects in the other-diagnosis subgroup and only 1 subject with suspected disk displacement without reduction who was dropped without further consideration. The data for mean age showed that the osteoarthritis subgroup (n = 12) was statistically older (17 years) than the disk condyle-incoordination-only subgroup (n = 11). The mean age of the other 2 groups, arthromyalgia (n = 11) and arthromyalgia with disk condyle incoordination (n = 6), was between the osteoarthritis and the disk-condyle-incoordination-only subgroups. For the 4 TMD subgroups whose data were analyzed, the mean differences between similar jaw opening measurements ranged from 6 to 8 mm with a standard deviation of approximately 8 to 10 mm. The mean left lateral motions were 0.5 to 1.3 mm larger than observed on the right. The widest mean jaw opening (56 mm) occurred in the disk-condyle-incoordination-only group. These differences were not found to be statistically significant. CONCLUSION: Analysis of opening, lateral and protrusive jaw motion data showed these measurements could not reliably differentiate between patients with osteoarthritis, arthromyalgia, arthromyalgia with disk condyle incoordination and disk condyle incoordination only. PMID- 12075205 TI - Caries experience and oral cleanliness in cystic fibrosis homozygotes and heterozygotes. AB - OBJECTIVES AND STUDY DESIGN: In the present study, the caries experience (ie, the number of decayed, missing, and filled teeth) and oral cleanliness (ie, measures of plaque, calculus, and gingival bleeding) of patients homozygous for cystic fibrosis (CF; n = 42), patients heterozygous for CF (n = 48), and healthy control subjects (n = 62) were recorded and statistically analyzed. RESULTS: Those who were homozygous for CF had a significantly lower caries experience than both the control subjects (P <.001) and those who were heterozygous for CF (P =.011). They also had significantly less gingival bleeding sites than the controls (P =.014) and those heterozygous for CF (P =.019). Patients heterozygous for CF showed significantly more extensive restorations than those who were homozygous (P =.015) and significantly more missing teeth than the controls (P =.008), whereas the controls had significantly more missing teeth than those who were homozygous for CF (P <.001). CONCLUSIONS: Although those homozygous for CF are expected to have a high caries risk because of their essential sugar-rich diet, they did not have a significantly higher caries experience in this study. Their better gingival health and lower caries experience may be attributed to medication use (antibiotics) and as-yet unidentified intrinsic salivary mechanisms. PMID- 12075206 TI - Oral manifestations of patients with Marfan syndrome: a case-control study. AB - OBJECTIVE: The purpose of this study was to conduct a complete analysis of the oral abnormalities of patients with Marfan syndrome. STUDY DESIGN: Twenty three patients with Marfan syndrome and 69 healthy controls were studied. The subjects were screened for cariologic and periodontal alterations, as well as structural defects of enamel and dentin. Data analysis was performed by using the t test, the chi-square test, and regression models. RESULTS: Patients aged 0 to 17 years were significantly at risk for caries. Local hypoplastic enamel spots were more frequent in Marfan syndrome and could be related to caries history of the deciduous dentition. Root deformity, abnormal pulp shape, and pulpal inclusions were a frequent finding in patients with Marfan syndrome. Calculus and gingival indices were significantly higher in the study group as well. CONCLUSIONS: This study shows the importance of early diagnosis of oral anomalies and timely treatment of dental problems in Marfan syndrome. A series of therapeutic guidelines to be integrated in treatment strategies is proposed. PMID- 12075207 TI - Immunohistochemical localization of growth factors fibroblast growth factor-1 and fibroblast growth factor-2 and receptors fibroblast growth factor receptor-2 and fibroblast growth factor receptor-3 in normal oral epithelium, epithelial dysplasias, and squamous cell carcinoma. AB - OBJECTIVES: Fibroblast growth factors (FGFs) and their receptors (FGFRs) have been identified in a variety of carcinomas, but there are few studies concerning their presence in oral cancers. The objective of this study was to determine whether FGF-1, FGF-2, and high affinity receptors FGFR2 and FGFR3 are present in the pathogenesis of oral epithelial dysplasias and oral squamous cell carcinoma. STUDY DESIGN: Sections from formalin-fixed, paraffin-embedded samples of oral normal mucosa (n = 14), epithelial dysplasia (n = 20), carcinoma in situ (n = 10), and squamous cell carcinoma (n = 12) were tested for cytoplasmic staining by standard in situ immunohistochemistry with antibodies for FGF-1, FGF-2, FGFR2, and FGFR3. RESULTS: Staining for FGF-1 is decreased or lost in the development of epithelial dysplasia and carcinoma. Staining for FGF-2 showed increased intensity (although not statistically significant) in oral epithelial dysplasias and squamous cell carcinomas and showed a significant increased expression in the upper layers of dysplasias and stratum spinosum-like cells in squamous cell carcinomas. Staining for FGFR2 showed a statistically significant increase in intensity in all layers of epithelial dysplasias and squamous cell carcinomas. Staining for FGFR3 was found in the upper stratum spinosum cells of normal and dysplastic epithelium and well-differentiated squamous cells in squamous cell carcinomas, with a statistically significant increase in staining intensity in dysplastic and carcinomatous tissues. CONCLUSIONS: The loss of FGF-1 is consistent with loss of differentiation in dysplasias and some squamous cell carcinomas. Changes in the localization of FGF-2 and FGFR2 into upper epithelial layers with increasing dysplasia suggest increased mitotic potential of high level cells. The co-localization of FGF-2 and its high affinity receptors in neoplastic tissues suggests an autocrine mechanism of influence on carcinogenesis. PMID- 12075208 TI - Rhabdomyosarcoma of the oral and maxillofacial region in Jordanians: a retrospective analysis. AB - OBJECTIVE: The objective was to study the clinicopathologic features of rhabdomyosarcoma (RMS) of the oral and maxillofacial region in Jordanians. STUDY DESIGN: Data were collected from records of patients treated between 1989 and 2000 at the Maxillofacial Unit of Jordan University of Science and Technology. The main outcome measures were age, gender, location, stage of disease, histopathologic type, treatment received, follow-up period, and eventual outcome. RESULTS: Nine patients with RMS aged 4 to 17 years were found, with a male to female ratio of 2:1. Six (67%) bony sites and 7 (78%) soft tissue sites were involved. The extent of disease was locoregional in 8 (89%) cases, nodal in 3 (33%) cases, and distant metastatic in 2 (22%) cases. The Intergroup Rhabdomyosarcoma Study classification of clinical groups was as follows: 2 (22%) cases in stage II, 5 (56%) cases in stage III, and 2 (22%) cases in stage IV. The histopathologic types found were 6 (67%) embryonal, 2 (22%) alveolar, and 1 (11%) undifferentiated. Triple agent chemotherapy was used in the treatment of 8 cases, singly or in combination with surgery or radiotherapy. Two patients are still alive, and 5 patients have died of disease. CONCLUSIONS: The clinicopathologic features of maxillofacial RMS in this group of Jordanians are different from those of people from other countries. Further studies are needed to have a better understanding of the behavior of RMS in the oral and maxillofacial area. PMID- 12075209 TI - Prevalence of Epstein-Barr virus in oral squamous cell carcinoma, oral lichen planus, and normal oral mucosa. AB - OBJECTIVE: The Epstein-Barr virus (EBV) is associated with both malignant and benign diseases in the head and neck region. In several studies it has also been associated with oral squamous cell carcinoma (OSCC). Oral lichen planus (OLP) is a disease with unknown origin, and viral antigens have been proposed as etiologic agents. Smoking and alcohol habits are known risk factors for oral cancer development. In this study, the prevalence of EBV in OSCC and OLP was investigated, along with the effect of smoking, alcohol use, and age on EBV prevalence. STUDY DESIGN: We examined 29 patients with OSCC, 23 with OLP, and 67 with clinically healthy oral mucosa. For EBV DNA analysis, a nested polymerase chain reaction method was used. RESULTS: The overall EBV prevalence in patients with oral disease was 32.1%. Of the OSCC patients, 37.9% were EBV positive; and of the OLP patients, 26.1% were EBV positive. Both percentages were statistically significant compared with that of control patients (7.3%). The difference in EBV prevalence between the smoking control group and the nonsmoking control group was insignificant. Increased age did not enhance EBV prevalence. CONCLUSIONS: This investigation shows that EBV is present in oral diseases such as OSCC and OLP. Smoking, alcohol use, or age does not seem to be a risk factor for EBV infection. The etiologic role of EBV in OSCC and OLP needs to be examined in a prospective follow-up study. PMID- 12075210 TI - Antimicrobial activity of extracts of calcium hydroxide points. AB - The purpose of this study was to evaluate in vitro the antibacterial effect of calcium hydroxide points by using a broth dilution test. Extracts of calcium hydroxide points and a calcium hydroxide paste (Calasept) were tested by using 2 facultative anaerobic bacteria, Streptococcus mutans (ATCC 27352) and Enterococcus faecalis (ATCC 29212). Saline solution was used as a control. Each tested agent was kept in contact with the bacterial species used for the experiment for 5 minutes, 1 hour, 1 day, 2 days, and 5 days. Results showed that Calasept paste was effective in killing the tested bacteria, whereas calcium hydroxide points and saline showed bacterial survival in all experimental periods. It was concluded that calcium hydroxide points extract is not an effective antimicrobial agent against the tested bacteria. PMID- 12075211 TI - Survival of first-time nonsurgical root canal treatment performed in a dental teaching hospital. AB - OBJECTIVE: The purpose of this study was to use survival analysis to evaluate the long-term treatment outcome of first-time, nonsurgical root canal therapy performed in a dental teaching hospital and to identify factors that might affect that outcome. STUDY DESIGN: One-tenth of all teeth treated between 1989 and 1994 were sampled. Patients were invited to return for a recall and were examined both clinically and radiographically. Any teeth that had been extracted or retreated or that were associated with a periapical radiolucency or presented with clinical signs and symptoms were deemed to have failed. An adjusted date of failure was calculated in the survival analysis by means of the Kaplan-Meier method. Ten covariables were further analyzed with the Cox regression model by using a backward stepwise regression method. RESULTS: Of 550 sampled teeth, 251 were examined. The mean observation period was 74 months. The failure rate was 44% (n = 111), with a median survival time of 113 months (mean, 91 months). Use of the Cox Regression model revealed that tooth type was significant in affecting the survival time of the treatment. Radiographic presence of voids in the apical and middle thirds of root canal fillings was associated with a significantly lower mean survival time than the presence of voids found in the coronal third or no void at all. The use of a calcium hydroxide dressing after initial treatment was associated with a longer survival time than failure to use medication or dressing with Ledermix alone. CONCLUSION: Simple calculation of percentages did not provide sufficient information on the treatment prognosis. The survival of first time root canal therapy was significantly influenced by the tooth type, radiographic location of voids in the root canal filling, and the intracanal medicament used. PMID- 12075212 TI - Postoperative pain after 1- and 2-visit root canal therapy. AB - OBJECTIVES: The factors that influence postoperative pain after root canal treatment are not completely understood. The purpose of this prospective clinical study was to evaluate postoperative pain after root canal therapy performed in 1 appointment versus 2 appointments. STUDY DESIGN: Seventy-two patients requiring root canal therapy on permanent molars were included in this study. Patients were randomly assigned to either the 1-appointment or the 2-appointment group. Both vital and nonvital teeth were included. The standardized protocol for all teeth involved local anesthesia, isolation and access, engine-driven rotary nickel titanium canal instrumentation to a minimum size #5 (.028 mm).04 taper Profile with step-back flaring, and irrigation with 2.5% NaOCl. Teeth in group 1 (n = 39) were obturated at the first appointment by using laterally condensed gutta-percha and Roth 811 sealer. Teeth in group 2 (n = 33) were closed with a sterile dry cotton pellet and Cavit restoration and were obturated at a second appointment 7 to 14 days later. A modified Visual Analogue Scale was used to measure preoperative pain and pain at 6, 12, 24, and 48 hours after the first appointment. Statistical analysis was performed to compare groups at each interval by using an independent-samples t test with Bonferroni adjustment. RESULTS: There was no statistically significant difference between groups at preoperative intervals or at any of the 4 postoperative intervals (P <.01). CONCLUSIONS: There was no difference in postoperative pain between patients treated in 1 appointment and patients treated in 2 appointments. The majority of patients in both groups reported no pain or only minimal pain within 24 to 48 hours of treatment. PMID- 12075213 TI - A comparison of the effectiveness of chloroform and eucalyptus oil in dissolving root canal sealers. AB - OBJECTIVE: The solubility of 8 different root canal sealers in chloroform and in eucalyptus oil was compared. STUDY DESIGN: For standardized samples (n=12), ring molds were filled with mixed sealers based on epoxy resin, silicone, calcium hydroxide, zinc oxide-eugenol, glass ionomer, and polyketone. These samples were immersed in chloroform or eucalyptus oil for 30 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, and 20 minutes. Then, the mean weight loss was determined and statistically analyzed. RESULTS: With the exception of the silicone, all the sealers showed significantly higher solubilities (P <.05) in chloroform than in eucalyptus oil. Epoxy resin was the most soluble sealer in chloroform. In eucalyptus oil, calcium hydroxide, and zinc oxide-eugenol showed the highest solubility. CONCLUSION: Under the conditions of this study, chloroform was a far more effective solvent of root canal sealers than eucalyptus oil. Because of the potential hazards of chloroform, further studies on the dissolution of root canal sealers in different solvents seem to be necessary. PMID- 12075214 TI - An effective method of inactivating chlorhexidine. AB - OBJECTIVE: The purpose of this study was to find an effective inactivating agent for chlorhexidine that would facilitate removal of all residual antimicrobial effect, which may cause false-negative results during microbiologic culturing. STUDY DESIGN: L-alpha-lecithin, Tween 80, and sodium thiosulfate were used in different proportions to prepare 6 potential inactivating solutions. Nine mL of each inactivating solution was mixed with 1 mL of 2% chlorhexidine solution. After 5 minutes of equilibration, 0.1 mL of bacterial cell suspension containing 2 x 10(4) viable cell of Enterococcus faecalis was added to the mixture. At 10 and 60 minutes, 0.1-mL aliquots were withdrawn and spread over blood agar plates and incubated at 37 degrees C for 72 hours. The number of colony-forming units on the blood agar plates was determined and recorded. RESULTS: The combination of 3% Tween 80 and 0.3% L-alpha-lecithin was found to be the most effective inactivating agent, allowing full recovery of the test organisms in the presence of chlorhexidine. CONCLUSION: The present study demonstrated a method to predictably inactivate chlorhexidine. PMID- 12075215 TI - Impact of lossy compression on diagnostic accuracy of radiographs for periapical lesions. AB - OBJECTIVES: The purpose of this study was to evaluate the lossy Joint Photographic Experts Group compression for endodontic pretreatment digital radiographs. STUDY DESIGN: Fifty clinical charge-coupled device-based, digital radiographs depicting periapical areas were selected. Each image was compressed at 2, 4, 8, 16, 32, 48, and 64 compression ratios. One root per image was marked for examination. Images were randomized and viewed by four clinical observers under standardized viewing conditions. Each observer read the image set three times, with at least two weeks between each reading. Three pre-selected sites per image (mesial, distal, apical) were scored on a five-scale score confidence scale. A panel of three examiners scored the uncompressed images, with a consensus score for each site. The consensus score was used as the baseline for assessing the impact of lossy compression on the diagnostic values of images. The mean absolute error between consensus and observer scores was computed for each observer, site, and reading session. RESULTS: Balanced one-way analysis of variance for all observers indicated that for compression ratios 48 and 64, there was significant difference between mean absolute error of uncompressed and compressed images (P <.05). After converting the five-scale score to two-level diagnostic values, the diagnostic accuracy was strongly correlated (R (2) = 0.91) with the compression ratio. CONCLUSION: The results of this study suggest that high compression ratios can have a severe impact on the diagnostic quality of the digital radiographs for detection of periapical lesions. PMID- 12075217 TI - Authorship, publication, and being a good doctor. PMID- 12075216 TI - Comparison of panoramic radiography and panoramic digital subtraction radiography in the detection of simulated osteophytic lesions of the mandibular condyle. AB - OBJECTIVE: The aim of this study was to compare panoramic temporomandibular joint radiography, unenhanced and color-enhanced digital-subtraction radiography with respect to detectability of simulated osteophytic lesions of the mandibular condyle. METHODS: Three dry human skulls with no obvious temporomandibular joint pathology were selected. Four sizes of bone chips were placed on the anterior aspect of the condyle at medial, central, and lateral locations. Panoramic radiographs were made with and without the chips in place. These paired radiographs were digitized, and unenhanced and color-enhanced digital-subtraction images of the original panoramic images were obtained. Eight observers evaluated 72 randomized images of each modality for the presence or absence of simulated osteophytic lesions of the mandibular condyle, grading the images on a 100-point scale. RESULTS: A(z) values for overall diagnostic accuracy of the three imaging modalities were 0.5376 for panoramic radiography, 0.7861 for unenhanced digital subtraction radiography, and 0.7923 for color-enhanced digital-subtraction radiography. Digital-subtraction radiography improved the detection accuracy of the original panoramic films. CONCLUSION: Panoramic radiographs were significantly less accurate in the detection of simulated osteophytic lesions of the mandibular condyle than the two digital-subtraction techniques. PMID- 12075218 TI - Botulinum toxin type B (MyoBloc) in the management of hyperkinetic facial lines. AB - OBJECTIVE: Our goal was to evaluate the safety and efficacy of botulinum toxin type B (BTX-B) in the management of hyperfunctional facial lines. STUDY DESIGN AND SETTING: Twenty-four patients from an academic faculty practice were treated with 400 to 800 units BTX-B in the corrugator, orbicularis oculi, or frontalis. Patients were evaluated with the Wrinkle Improvement Score (WIS) and Rated Numeric Kinetic Line Scale (RNKLS) and questioned about side effects and pain. Evaluations were made at 1, 2, 4, 8, and 12 weeks. RESULTS: Onset of effect was within 72 hours. WIS and RNKLS for all sites were statistically better after treatment, with the effect lasting 8 weeks. In general, patients noted a moderate improvement (grade 2) on WIS and a 2-point improvement on RNKLS. Injection was more painful than that of BTX-A. There were no complications. CONCLUSION: BTX-B is a safe and effective technique to eliminate wrinkles. Further studies are needed to elucidate dosage and duration. SIGNIFICANCE: BTX-B is stable for prolonged periods of time, requires no preparation before use, and is antigenetically distinct from BTX-A. PMID- 12075219 TI - Mitomycin C for the prevention of adhesion formation after endoscopic sinus surgery: a randomized, controlled study. AB - OBJECTIVE: Mitomycin C (MMC) is an antineoplastic agent that has been shown to decrease scar tissue after ophthalmologic surgery. Our goal was to determine whether the application of MMC at the conclusion of sinus surgery decreases the incidence of postoperative adhesion formation. METHODS: At the completion of endoscopic sinus surgery in 55 patients, a cotton pledget saturated with 1 mL of 0.4 mg/mL MMC was placed for 4 minutes in the right or left middle meatus and a similar saline-soaked pledget was placed on the opposite side. Patients were examined postoperatively by a masked observer for the presence of synechiae and mucosal changes. RESULTS: Postoperative adhesions were observed in 16 patients (29%) with a mean follow-up of 4.1 months. These adhesions were bilateral in 6 patients (10.9%) and unilateral in 10 patients (18%). Unilateral adhesions were observed on only 2 sides (3.6%) treated with MMC and 8 controls (14.5%) (P = 0.058). No adverse effects were observed. CONCLUSIONS: MMC was found to be safe to use during sinus surgery, and it may reduce the incidence of postoperative adhesions at the dosage used in this study. SIGNIFICANCE: Because of the observed trend toward decreased synechiae formation with MMC application, further trials using higher concentrations and application times are warranted. PMID- 12075220 TI - Acoustic rhinometry findings in patients with mild sleep apnea. AB - BACKGROUND: Nasal obstruction may contribute to the development of obstructive sleep apnea (OSA). Acoustic rhinometry (AR) measures nasal patency and congestion, which are useful parameters in objectively evaluating nasal obstruction. The nasal obstruction produced by allergic rhinitis may contribute to the development of OSA and can be easily assessed with AR. OBJECTIVE: This study was undertaken to assess the degree of nasal obstruction seen in allergic patients with and without OSA. STUDY DESIGN AND SETTING: This study was a retrospective data analysis from a tertiary referral center. The AR data from 10 patients with and 40 patients without mild OSA were compared. RESULTS: The mean congestion factors at the first cross-sectional area (CSA1) on the AR graph were found to be significantly higher in the OSA group than in the non-OSA group (P = 0.03). The classification of change in congestion factors demonstrated significant differences at CSA1, CSA2, and CSA3 and in volume (t distributions <0.001, 0.0312, <0.001, and <0.001, respectively). The non-OSA patients noted a significant subjective improvement in nasal congestion after topical nasal decongestion, whereas the OSA patients did not (P 12 years. The subjects of this study were matched based on socioeconomic status, residual hearing before cochlear implantation, the kinds of cochlear implant device, speech processing strategy, communication mode after implantation, and primary language in family. All of them have used the device minimally for 2 years. The subjects were tested with a range of closed and open-set auditory speech perception tests, and the levels of auditory speech perception in different age groups were compared. Results showed that the children who received an implant at 0 to 3 years of age had maximum auditory speech perception. PMID- 12075228 TI - Biomechanical effects of hydration in vocal fold tissues. AB - OBJECTIVE: It has often been hypothesized, with little empirical support, that vocal fold hydration affects voice production by mediating changes in vocal fold tissue rheology. To test this hypothesis, we attempted in this study to quantify the effects of hydration on the viscoelastic shear properties of vocal fold tissues in vitro. STUDY DESIGN: Osmotic changes in hydration (dehydration and rehydration) of 5 excised canine larynges were induced by sequential incubation of the tissues in isotonic, hypertonic, and hypotonic solutions. Elastic shear modulus (G'), dynamic viscosity eta' and the damping ratio zeta of the vocal fold mucosa (lamina propria) were measured as a function of frequency (0.01 to 15 Hz) with a torsional rheometer. RESULTS: Vocal fold tissue stiffness (G') and viscosity (eta) increased significantly (by 4 to 7 times) with the osmotically induced dehydration, whereas they decreased by 22% to 38% on the induced rehydration. Damping ratio (zeta) also increased with dehydration and decreased with rehydration, but the detected differences were not statistically significant at all frequencies. CONCLUSION: These findings support the long-standing hypothesis that hydration affects vocal fold vibration by altering tissue rheologic (or viscoelastic) properties. SIGNIFICANCE: Our results demonstrated the biomechanical importance of hydration in vocal fold tissues and suggested that hydration approaches may potentially improve the biomechanics of phonation in vocal fold lesions involving disordered fluid balance. PMID- 12075229 TI - The pedicled superficial temporalis fascial flap: a new method for reconstruction in otologic surgery. AB - OBJECTIVES: Our goal was to describe a novel fascial flap of the temporal region and its use for reconstruction in otologic and neurotologic surgery. METHODS: The superficial temporalis fascia has an axial blood supply derived from the middle temporal artery and can be raised independently from the overlying temporoparietal fascia or the underlying deep temporalis fascia. This flap was used on 15 consecutive patients to solve a wide variety of reconstructive problems after otologic procedures. RESULTS: No additional morbidity was observed from the use of this flap. There were no complications related to the reconstruction. Adequate exposure for raising this flap was obtained using standard incisions for the otologic procedures. Follow-up ranges from 2 to 25 months. CONCLUSIONS: This fascial flap provides a wide surface area of tissue on a narrow-based pedicle capable of a wide arc of rotation. It provides thin, pliable tissue that can be adapted to the needs of various reconstructive otologic/neurotologic problems. PMID- 12075230 TI - Promoter hypermethylation: an important epigenetic mechanism for hMLH1 gene inactivation in head and neck squamous cell carcinoma. AB - OBJECTIVE: The hMLH1 gene is one of the mismatch DNA repair genes. Inactivation of the hMLH1 gene has been implicated in the tumorigenesis of many types of human cancers. In most sporadic forms of human cancers, promoter hypermethylation is responsible for hMLH1 gene inactivation. Lack of hMLH1 protein expression has been found in a subset of head and neck squamous cell carcinomas (HNSCCs). The purpose of this study was to investigate whether promoter hypermethylation causes hMLH1 gene inactivation in HNSCCs. STUDY DESIGN: hMLH1 protein expression was determined by immunohistochemical staining in 62 cases, whereas hMLH1 gene promoter methylation was analyzed by methylation-sensitive restriction enzyme digestion, followed by polymerase chain reaction, in 35 cases of HNSCCs. RESULTS: Sixteen (26%) of 62 cases of HNSCCs showed near-complete loss of hMLH1 protein expression on immunohistochemical staining. Twelve (92%) of 13 cases that were negative for the hMLH1 protein displayed promoter hypermethylation, whereas 17 (77%) of 22 cases positive for the protein were free of promoter methylation. CONCLUSIONS: Promoter hypermethylation may be an important mechanism for hMLH1 gene inactivation in a subset of HNSCCs. PMID- 12075231 TI - Comparison of ThinPrep versus conventional smear cytopreparatory techniques for fine-needle aspiration specimens of head and neck masses. AB - OBJECTIVES: Diagnostic accuracy of the ThinPrep process (Cytyc, Boxborough, MA) was compared with that of conventional (smear) cytopreparation for fine-needle aspiration (FNA) of head and neck masses. METHODS: In a prospective, randomized, single-blinded study, 209 patients served as their own controls and underwent 236 FNAs using ThinPrep and conventional (smear) cytopreparatory techniques. RESULTS: ThinPrep produced less air-drying artifact and less mechanical distortion than the conventional method. The conventional technique was diagnostic in 63% of samples; the ThinPrep technique was diagnostic in 55% of samples. When all results were combined, pathologists subjectively preferred the conventional technique but accepted use of ThinPrep as the only cytopreparatory technique for most head and neck masses. CONCLUSIONS: For adequately experienced cytopathologists, ThinPrep is acceptable for FNA of salivary masses, neck cysts, metastatic lymph nodes, and thyroid lesions. Conventional smear technique should be used for FNA of nonmetastatic lymphoid lesions. Use of ThinPrep can complement use of the conventional (smear) cytopreparatory technique when aspirate is nondiagnostic or bloody, when the patient has a blood-borne infectious disease, when the clinician is inexperienced, or when aspirate has entered the syringe. PMID- 12075232 TI - A review of outcome measurements for voice disorders. AB - There has been great interest in the medical and associated communities in the development and use of patient-based outcome measures such as quality-of-life and handicap indexes. This area of assessment of initial disability and response to subsequent treatment is especially applicable to the field of laryngology and voice disorders given the limitations of perceptual analysis and objective vocal function studies. In this review, we outline the development and importance of patient-based outcome measures in general, and their application for voice disorders specifically. In addition, several voice-specific instruments are discussed in detail and provided. One of the primary goals of this review is to educate and facilitate the future use of these simple but important patient-based outcome instruments for otolaryngologists who care for patients with voice disorders. PMID- 12075233 TI - World Wide Web-based temporal bone anatomic sections. AB - The study goal was to post a complete set of temporal bone anatomy slides on the World Wide Web for anatomic study. Magnification of the width of the sections varied from 2.4 to 33.6 inches. Good representation of gross and tissue anatomy was obtained, but cellular detail was lacking. The website used approximately 37 megs of disk space. The resultant website provides a cost-effective method of presenting large color photographs of temporal bone anatomy. Thumbnails allow comparison between sections, which aids in learning 3-dimensional anatomy. A method for measuring distance on digitized images is also described. The sections may be viewed at www.ear-anatomy.com, and continuing medical education credits may be earned at www.cme-usa.org. PMID- 12075234 TI - A parathyroid cyst with mediastinal extension: report of a case. PMID- 12075235 TI - Tuberculous parotitis: a forgotten entity. PMID- 12075236 TI - Unilateral supraglottic edema after the use of a laryngeal mask airway. PMID- 12075237 TI - Psammomatoid ossifying fibroma of the temporal bone. PMID- 12075238 TI - Frontal sinus mucocele presenting as oculomotor nerve palsy. PMID- 12075239 TI - Nasal granuloma gravidarum. PMID- 12075240 TI - No variants in the cardiac actin gene in Finnish patients with dilated or hypertrophic cardiomyopathy. AB - BACKGROUND: Dilated and hypertrophic cardiomyopathies are primary myocardial diseases that cause considerable morbidity and mortality. Although these cardiomyopathies are clinically heterogeneous, genetic factors play an important role in their etiology and pathogenesis. The defects in the cardiac actin (ACTC) gene can cause both cardiomyopathies. The aim of our study was to screen for variants in the ACTC gene in patients with dilated or hypertrophic cardiomyopathy from Eastern Finland. MATERIALS AND METHODS: Altogether, 32 patients with dilated and 40 patients with hypertrophic cardiomyopathy were included in the study. Commonly approved diagnostic criteria were applied, and secondary cardiomyopathies were carefully excluded. All 6 exons of the ACTC gene were amplified with polymerase chain reaction and screened for variants with single strand conformation polymorphism analysis. RESULTS AND CONCLUSION: We did not find any new or previously reported variants. Our results indicate that defects in the ACTC gene do not explain dilated cardiomyopathy or hypertrophic cardiomyopathy in subjects from Eastern Finland and confirm earlier results that the ACTC gene does not play an important role in the genetics of dilated or hypertrophic cardiomyopathies. PMID- 12075241 TI - Determination of in vivo velocity and endothelial shear stress patterns with phasic flow in human coronary arteries: a methodology to predict progression of coronary atherosclerosis. AB - BACKGROUND: Although the coronary arteries are equally exposed to systemic risk factors, coronary atherosclerosis is focal and eccentric, and each lesion evolves in an independent manner. Variations in shear stress elicit markedly different humoral, metabolic, and structural responses in endothelial cells. Areas of low shear stress promote atherosclerosis, whereas areas of high shear stress prevent atherosclerosis. Characterization of the shear stresses affecting coronary arteries in humans in vivo may permit prediction of progression of coronary disease, prediction of which plaques might become vulnerable to rupture, and prediction of sites of restenosis after percutaneous coronary intervention. METHODS: To determine endothelial shear stress, the 3-dimensional anatomy of a segment of the right coronary artery was determined immediately after directional atherectomy by use of a combination of intracoronary ultrasound and biplane coronary angiography. The geometry of the segment was represented in curvilinear coordinates and a computational fluid dynamics technique was used to investigate the detailed phasic velocity profile and shear stress distribution. The results were analyzed with several conventional indicators and one novel indicator of disturbed flow. RESULTS: Our methodology identified areas of minor flow reversals, significant swirling, and large variations of local velocity and shear stress--temporally, axially, and cirumferentially--within the artery, even in the absence of significant luminal obstruction. CONCLUSIONS: We have described a system that permits, for the first time, the in vivo determination of pulsatile local velocity patterns and endothelial shear stress in the human coronary arteries. The flow phenomena exhibit characteristics consistent with the focal nature of atherogenesis and restenosis. PMID- 12075242 TI - Routine statin treatment after acute coronary syndromes? PMID- 12075243 TI - An interventional cardiologist: a cardiologist who intervenes. PMID- 12075244 TI - Recertification: mandatory or voluntary? Teaching old docs new tricks. PMID- 12075245 TI - Recertification: nobody asked me! PMID- 12075246 TI - Two vessels or not two vessels? That is the question. PMID- 12075247 TI - Making positive out of negative trials. PMID- 12075248 TI - The SYNERGY trial: study design and rationale. AB - BACKGROUND: Enoxaparin was shown to be superior to unfractionated heparin in the patients with non-ST-segment elevation acute coronary syndromes (ACS) in the Efficacy and Safety of Subcutaneous Enoxaparin in Non-Q-wave Coronary Events study and the Thrombolysis In Myocardial Infarction (TIMI) 11B trial. However, enoxaparin has had limited acceptance in clinical practice, in part because of the contemporary management of these patients, which includes glycoprotein IIb/IIIa inhibition and the use of early invasive management strategies. STUDY DESIGN: The Superior Yield of the New strategy of Enoxaparin, Revascularization and GlYcoprotein IIb/IIIa inhibitors (SYNERGY) trial is an 8000-patient, prospective, randomized, open-label, multicenter investigation of enoxaparin compared with unfractionated heparin in patients at high risk with non-ST-segment elevation ACS treated with an early invasive strategy. The primary efficacy end point is death or nonfatal myocardial infarction 30 days after enrollment. IMPLICATIONS: The SYNERGY trial is the largest study currently planned for the acute therapy of patients with non-ST-segment elevation ACS and the first large trial since the publication of the revised American College of Cardiology/American Heart Association guidelines for the management of these patients. In addition to evaluating the potential superiority of enoxaparin over unfractionated heparin, this investigation will provide important observations of current treatment strategies in patients with ACS. PMID- 12075249 TI - Prevalence and clinical correlates of peripheral arterial disease in the Framingham Offspring Study. AB - BACKGROUND: Peripheral arterial disease (PAD) is associated with an increased risk for mortality. We sought to assess the prevalence of PAD and its risk factors in a population-based sample. METHODS: We examined 1554 males and 1759 females with a mean age of 59 years who attended a Framingham Offspring Study examination from 1995 to 1998. PAD was defined by an ankle-brachial blood pressure index of <0.9. Age- and sex-adjusted and multivariable logistic regression analyses were performed to identify factors associated with PAD. RESULTS: The prevalences of PAD, current intermittent claudication, lower extremity bruits and surgical intervention were 3.9%, 1.9%, 2.4% and 1.4% in males and 3.3%, 0.8%, 2.3% and 0.5% in females. Hypercholesterolemia, high density lipoprotein cholesterol, triglyceride, diabetes, hypertension, current smoking, pack-years of smoking, body mass index, fibrinogen, and prevalent coronary disease were associated with PAD in age- and sex-adjusted analyses. Odds ratios and 95% CIs for significant associations identified from multivariable analyses are as follows: each 10 years of age, 2.6 (2.0, 3.4); hypertension, 2.2 (1.4, 3.5); smoking, 2.0 (1.1, 3.4); 10 pack-years of smoking, 1.3 (1.2, 1.4); 50 mg/dL of fibrinogen, 1.2 (1.1, 1.4); 5 mg/dL of high-density lipoprotein, 0.9 (0.8, 1.0); coronary disease, 2.6 (1.6, 4.1). CONCLUSIONS: Smoking cessation and hypertension control are important goals in the aim to reduce PAD and its associated impact on quality of life, functional decline, and risk for subsequent cardiovascular disease. PMID- 12075250 TI - Identification of patients at high risk for death and cardiac ischemic events after hospital discharge. AB - BACKGROUND: Patients with unstable angina (UA) and non-ST-elevation myocardial infarction (NSTEMI) remain at risk for death and cardiac ischemic events after being discharged from the hospital. METHODS: We examined whether the Thrombolysis In Myocardial Infarction (TIMI) risk score for UA/NSTEMI, ascertained at presentation in patients enrolled in the TIMI 11B and Efficacy and Safety of Subcutaneous Enoxaparin in Unstable Angina and Non-Q-Wave MI (ESSENCE) trials, could be used to identify patients at high risk for major cardiac events after hospital discharge. RESULTS: There were a total of 1218 major cardiac events, defined as death, nonfatal myocardial infarction, or urgent revascularization, by day 43. Of these events, 336 (28%) occurred in patients after they were discharged from the hospital. Use of the TIMI risk score for UA/NSTEMI revealed a progressive, statistically significant increase in the rate of events after leaving the hospital as the patients' baseline level of risk increased (P <.001 for chi(2) test for trend). For patients with a risk score of 5 to 7, treatment with enoxaparin during the acute phase was associated with an odds ratio of 0.51 (95% CI 0.29-0.91) for the occurrence of death and cardiac ischemic events after hospital discharge. CONCLUSIONS: More than one fourth of the major cardiac events that will occur in the first 6 weeks occur after discharge from the hospital. Stratification at presentation on the basis of the TIMI risk score for UA/NSTEMI can be used to identify patients at high risk for these events. Among patients at high-risk, acute-phase treatment with enoxaparin significantly reduces the risk of major cardiac events after leaving the hospital. PMID- 12075251 TI - CD14 gene -159C/T polymorphism is not associated with coronary artery disease and myocardial infarction. AB - BACKGROUND: Monocyte differentiation antigen CD14 is considered an important cell activating mediator of inflammatory responses that may result in atherosclerosis, coronary artery disease (CAD), thrombus formation, and myocardial infarction (MI). We assessed the possibility that a C --> T nucleotide substitution polymorphism in the promoter (position -159) of the gene encoding CD14 constitutes a risk factor for CAD and MI. METHODS: Consecutive patients with significant, angiographically documented coronary stenoses but without symptoms or signs of old or acute MI constituted the group with CAD (n = 998). Consecutive patients with angiographic examination with old or acute MI constituted the group with MI (n = 793). Subjects matched with patients for age and gender but without angiographic evidence of CAD and without symptoms or signs of MI (n = 340) and a group of healthy blood donors (n = 104) served as controls. RESULTS: Genotype distributions of the -159C/T polymorphism were similar across the groups; CC:CT:TT was 26.9%:51.0%:22.1% in blood donors, 25.9%:52.0%:22.1% in matched control subjects, 27.4%:49.9%:22.7% in patients with CAD, and 29.2%:49.2%:21.6% in patients with MI. The lack of association persisted also after adjustment for the presence of conventional cardiovascular risk factors. In addition, no significant differences were found between genotype distributions of control subjects and selected subgroups of patients with CAD or MI. CONCLUSION: These findings indicate that, in the sample of patients examined in this study, the 159C/T polymorphism of the CD14 gene is not related to CAD or MI. PMID- 12075252 TI - Effects of cardiac rehabilitation and exercise training on autonomic regulation in patients with coronary artery disease. AB - BACKGROUND: Although cardiovascular rehabilitation and exercise training have substantial benefits in various ischemic heart disease (IHD) risk factors and subsequent prognosis after major IHD events, there is a paucity of information about its effects on autonomic regulation (such as heart rate variability [HRV] and baroreflex gain), particularly considering its arterial and cardiopulmonary components. METHODS: We studied 40 patients (aged 60 +/- 6 y) after major IHD events, including 29 who underwent a comprehensive phase II cardiac rehabilitation and exercise training program and 11 controls who did not attend cardiac rehabilitation. Specifically, we determined whether active training improves prognostic indices of autonomic regulation of the SA node and whether changes in baroreflex gain could be ascribed to the arterial or to the cardiopulmonary component of the overall arterial pressure/heart period baroreflex. RESULTS: Only patients with IHD undergoing active rehabilitation demonstrated a significant increase in R-R interval, in its variance, in overall gain of arterial pressure/heart period baroreflex (7.44 +/- 1.20 ms/mm Hg to 12.12 +/- 1.48 ms/mm Hg, P <.001) and in peak oxygen consumption (Delta = 2.45 mL/kg/min, P <.001). Separate examination of the selective arterial and cardiopulmonary components showed that only the latter increased significantly (6.17 +/- 1.09 ms/mm Hg to 10.62 +/- 1.56 ms/mm Hg; P <.01). CONCLUSIONS: Cardiac rehabilitation is associated with significant improvements in autonomic markers of neural regulation of the SA node, such as increases in R-R variance and the gain of the overall spontaneous baroreflex, with specific improvements in the cardiopulmonary component as opposed to the arterial baroreflex component of this system. These improvements may further explain the reduction in morbidity and mortality noted after formal cardiac rehabilitation and exercise training programs. PMID- 12075253 TI - Quality of life improves with treatment in the Canadian Trial of Atrial Fibrillation. AB - BACKGROUND: The impact of atrial fibrillation (AF) and its treatment on health related quality of life (QOL) is not well understood. We assessed QOL in patients with symptomatic AF participating in the Canadian Trial of Atrial Fibrillation. METHODS: Self-report QOL questionnaires including the Short-Form-36 (SF-36), symptom checklist (SCL) and AF Severity Scale (AFSS) were completed at baseline and 3, and 12 months after randomization. RESULTS: The study group was aged 65 +/ 10 years and 59% were male. By design, 50% of patients were randomized to amiodarone (n = 132), 25% to sotalol (n = 66), and 25% to propafenone (n = 66). Most patients had normal left ventricular function (89%). Physical (41.9 +/- 9.3 to 43.7 +/- 9.2, P =.001) and mental health (47.5 +/- 10.5 to 49.0 +/- 9.8, P =.023) summary measures from the SF-36 improved significantly from baseline to 3 months. Arrhythmia symptom frequency and severity (SCL) also improved markedly from baseline to 3 months (symptom frequency 20.4 +/- 9.4 to 16.2 +/- 9.5 and symptom severity 16.7 +/- 8.2 to 12.9 +/- 7.4, both P <.001). QOL improvements were not significantly different among the groups randomized to amiodarone, sotalol, or propafenone. However, patients with no symptomatic recurrences of AF had higher scores at 3 months on measures of global well-being than those with recurrences in the first 3 months (7.4 +/- 1.8 vs 6.9 +/- 1.8, P <.05). There were no significant QOL changes from the 3 to 12 month assessment. CONCLUSION: In patients with symptomatic AF, QOL improves after treatment, independent of the specific drug used for treatment. This is especially true for patients in whom treatment prevents AF recurrence. PMID- 12075254 TI - Baseline characteristics of patients with atrial fibrillation: the AFFIRM Study. AB - BACKGROUND: Although anticoagulation therapy is accepted for most patients with atrial fibrillation, 2 different strategies exist for management of the cardiac rhythm: atrial fibrillation is allowed to persist while the ventricular rate is controlled; and atrial fibrillation is converted, and an attempt is made to maintain sinus rhythm. METHODS: The Atrial Fibrillation Follow-up Investigation of Rhythm Management (AFFIRM) Study was a randomized clinical trial that compared these 2 strategies. We report the baseline characteristics of the patients enrolled in the AFFIRM Study. RESULTS: More than 7400 patients at more than 200 North American hospitals and clinics qualified for enrollment in the AFFIRM Study. A total of 4060 patients were enrolled in the AFFIRM Study. The average age of patients enrolled was 70 years, with 39% female and 89% white. Hypertension was present in 71%. Coronary artery disease was present in 38%. Echocardiography was performed in 3311 patients, and results showed normal ventricular function in 68% and normal left atrial size in 33%. Most patients with recurrent episodes had symptoms with atrial fibrillation. Approximately one third of patients were enrolled with a first episode of atrial fibrillation. CONCLUSION: The AFFIRM Study enrolled 4060 predominantly elderly patients with atrial fibrillation to compare ventricular rate control with rhythm control. The patients in the AFFIRM Study were representative of patients at high risk for complications from atrial fibrillation, which indicates that the results of this large clinical trial will be relevant to patient care. PMID- 12075255 TI - Intrauterine diagnosis of heterotaxy syndrome. AB - BACKGROUND: Heterotaxy syndrome, including right isomerism and left isomerism, is characterized by an abnormal symmetry of the viscera and veins and is frequently associated with complex cardiac anomalies. We sought to define the feasibility of in utero diagnosis and the postnatal outcome. METHODS: Patients with heterotaxy syndrome were identified from 579 fetal echocardiograms performed from January 1994 to December 1998. The diagnosis was made on the basis of the fetal echocardiographic findings and was confirmed with autopsy or postnatal evaluation. RESULTS: A total of 25 fetuses with right isomerism and 4 with left isomerism constitute the study population. The pregnancies of 7 fetuses (6 right and 1 left isomerism) were terminated before the 24th gestational week and subjected to autopsy. Twelve fetuses (10 right and 2 left isomerism) were lost to follow-up. Nine with right isomerism and 1 with left isomerism were delivered and underwent palliation. Among them, 5 patients (56%) with right isomerism died and more than half of the deaths occurred during infancy. The major cardiac anomalies detected and confirmed with postnatal evaluation or autopsy in fetuses with right isomerism were total anomalous pulmonary venous connection (6/15; 40%), common atrium (15/15; 100%), complete atrioventricular canal (15/15; 100%), double outlet right ventricle (15/15; 100%), and pulmonary stenosis (11/15; 73%). The major cardiac anomalies in fetuses with left isomerism were interruption of inferior vena cava (2/2; 100%), common atrium (1/2; 50%), and complete atrioventricular canal (1/2; 50%). Undetected lesions with fetal echocardiogram were abnormal pulmonary venous return to systemic veins in 1 case (sensitivity, 83%; 5/6; and specificity, 90%; 9/10) and outflow obstruction in 1 case (sensitivity, 91%; 11/12; and specificity, 67%; 2/3). Different patterns of rhythm disturbances were identified: supraventricular tachycardia in 1 case with right isomerism and sinus bradycardia with junctional rhythm in 3 cases with left isomerism (2 of them lost to follow-up). After birth, another 2 patients with right isomerism had supraventricular tachycardia, and 1 with left isomerism had sinus bradycardia develop at age 2 years. CONCLUSION: Heterotaxy syndrome is usually detected in fetuses with the sonographic cardiac abnormalities. Visualization of the pulmonary venous return and outflow obstruction and characterization of the rhythm disturbances are feasible. However, in spite of prenatal diagnosis, the prognosis remains poor. PMID- 12075256 TI - Low settings of the ventricular pacing output in patients dependent on a pacemaker: are they really safe? AB - BACKGROUND: It is generally acknowledged that pacemaker output must be adjusted with a 100% voltage safety margin above the pacing threshold to avoid ineffective pacing, especially in patients dependent on pacemakers. AIMS: The aim of this prospective crossover study was to assess the beat-to-beat safety of low outputs in patients who are dependent on a pacemaker between 2 follow-up examinations. METHODS: The study included 12 patients who had received a DDD pacemaker with an automatic beat-to-beat capture verification function. The ventricular output at 0.4 milliseconds pulse duration was programmed independently of the actual pacing threshold in a crossover randomization to 1.0 V, 1.5 V, and 2.5 V for 6 weeks each. At each follow-up, the diagnostic counters were interrogated and the pacing threshold at 0.4 milliseconds was determined in 0.1-V steps. The diagnostic pacemaker counters depict the frequency of back-up pulses delivered because of a loss of capture. During the randomization to 1.0-V output, we evaluated whether the adjustment of the output under consideration of the >100% voltage safety margin reduced the frequency of back-up pulses. RESULTS: Pacing thresholds at the randomization to 1.0-V, 1.5-V, and 2.5-V output were not significantly different, with 0.7 +/- 0.3 V at 2.5-V output, 0.6 +/- 0.2 V at 1.5-V output, and 0.6 +/- 0.2 V at 1.0-V output. The frequency of back-up pulses was similar at 2.5-V and 1.5-V output, 2.2% +/- 1.9% and 2.0% +/- 2.0%, respectively. The frequency of back-up pulses significantly increased at 1.0-V output to 5.8% +/- 6.4% (P <.05). Back-up pulses >5% of the time between the 2 follow-ups were observed in no patient at 2.5 V, in 1 patient at 1.5 V, and in 5 patients at 1.0 V. At the randomization to the 1.0-V output, 6 patients had pacing thresholds of 0.5 V or less, and 6 patients had pacing thresholds >0.5 V. The frequency of back-up pulses in the 2 groups was not significantly different, 6.4% +/- 8.6% and 5.7% +/ 2.6%. CONCLUSIONS: The frequency of back-up pulses was significantly higher at 1.0-V output than at 1.5-V and 2.5-V output. This also applied to patients with pacing thresholds of < or =0.5 V. Fixed low outputs seem not to be absolutely safe between 2 follow-ups in patients who are dependent on a pacemaker, even when the output has a 100% voltage safety margin above the pacing threshold. When patients with pacemakers programmed to a low ventricular output have symptoms of ineffective pacing, an intermittent increase of the pacing threshold should be carefully ruled out. PMID- 12075257 TI - Transmyocardial laser revascularization and left ventricular reduction surgery affect ventricular arrhythmias and heart rate variability. AB - BACKGROUND: Transmyocardial laser revascularization (TMLR) and left ventricular reduction by endoventricular patch plasty (LVR) are 2 new surgical procedures performed in patients with endstage coronary artery disease and left ventricular dilation/aneurysms, respectively. As these are performed in patients at high risk for sudden cardiac death and may interact with arrhythmogenesis, we assessed the influence of these procedures on incidence and severity of ventricular tachyarrhythmias and time-domain heart rate variability. METHODS: Preoperative and one week postoperative 24-hour Holter recordings were performed in 37 patients undergoing TMLR (n = 23, CO2-laser technique) or LVR (n = 14). RESULTS: TMLR patients received a mean of 27.2 +/- 9.2 laser channels. Postoperatively, the proportion of patients who underwent TMLR with spontaneous ventricular tachycardia (> or =4 repetitive ventricular beats) increased (0% vs 26%, P <.05), including one patient who died from documented ventricular fibrillation during monitoring. There was no correlation to the number and/or location of laser induced channels or to perioperative CK levels. HRV parameters were not altered by TMLR. By contrast, LVR did not significantly influence ventricular tachyarrhythmia episodes but markedly depressed all major HRV parameters (SDNN 116.4 vs 61.8, RMSSD 35.2 vs 19.9, pNN50 14.5 vs 4.9, all P <.05). CONCLUSIONS: Early after TMLR, there is evidence of an increased incidence of spontaneous ventricular tachycardia enhancing the risk for sudden cardiac death, while HRV remains unaffected. By contrast, LVR resulted in a marked reduction in HRV still present one week postoperatively, while no effect was observed on incidence and/or severity of spontaneous ventricular tachyarrhythmias. PMID- 12075258 TI - Staged versus one-step approach for multivessel percutaneous coronary interventions. AB - BACKGROUND: Percutaneous coronary interventions (PCIs) in patients with multivessel coronary artery disease (CAD) may be staged or performed in a single session. No data exist about the relative safety and efficacy of these 2 strategies. Our aim was to compare short-term and long-term outcomes of patients with multivessel CAD who underwent PCI in 1 versus 2 sessions. METHODS AND RESULTS: The study included 264 consecutive patients who underwent treatment in our center during 1997 and 1998. PCI was conducted in a single session in 129 patients and was staged in 135 patients. The mean interval between the sessions in the staged group was 45.6 +/- 22.3 days. The rates of major adverse cardiac events (MACEs) during in-hospital stay did not differ significantly between the staged (combined for both stages) and nonstaged groups (2.2% vs 4.6%; P =.28). A trend for lower event rates at 30-day (2.9% vs 6.9%; P =.13) and 1-year follow-up (26.1 vs 35.9; P =.08) favored the staged arm. Diameter stenosis > or =50% was found in 17% of patients in the staged group in the second session and was successfully retreated in most of them. No MACE occurred between the sessions. Multivariate analysis identified staging of the procedure as a single independent predictor of MACE at 1-year follow-up (P =.05). CONCLUSION: Our results suggest that a practical staging strategy within 4 to 8 weeks is safe and allows for identification and treatment of potential excessive proliferative response in the previously intervened lesions during the second procedure. PMID- 12075259 TI - Could direct stenting reduce no-reflow in acute coronary syndromes? A randomized pilot study. AB - OBJECTIVES: Recently, direct stenting has been shown in retrospective and randomized studies to be feasible and safe in highly selected patients, with a potential interest to reduce the cost of the procedure and the rate of no-reflow. This randomized pilot study was designed to compare the incidence of no-reflow after direct stenting or conventional stenting after balloon predilation in acute coronary syndrome-related lesions. METHODS AND RESULTS: Between December 1998 and October 1999, 130 patients in our center with acute coronary syndromes were included in this study and randomized in 2 groups. In group A (n = 65), direct stent implantation was performed without balloon predilation. In group B (n = 65), conventional balloon predilation was carried out before stent implantation. Baseline clinical and angiographic characteristics before the procedure were similar in the 2 groups of patients. No-reflow was observed in 7.7% after direct stenting and in 6.1% after conventional stent implantation (P = not significant). The immediate clinical success rate was similar in the 2 groups. Among the procedural data, only duration of the procedure (shorter in the direct stenting group), the number of balloons used, and the quantity of contrast agent (lower in the direct stenting group) were significantly different between the 2 groups (P <.05). The 6-month clinical outcome was similar in the 2 groups. CONCLUSION: This randomized study confirms the promising results of previous studies that show the feasibility and the safety of direct coronary stenting in highly selected acute coronary syndrome-related lesions. The major impact of this strategy is the improvement of the cost-benefit ratio, with no major influence on the acute complications and especially on the occurrence of no-reflow in this high-risk population. PMID- 12075260 TI - Use of the invasive management strategy for patients with non-Q-wave myocardial infarction: an observational database report from the Worcester Heart Attack Study. AB - BACKGROUND: Recent randomized clinical trials have suggested a benefit of an invasive management strategy in patients with acute coronary syndromes. However, the broader use and impact of the invasive management approach has not been established for patients with acute coronary syndromes beyond the relatively narrow patient populations studied in randomized, clinical trials. METHODS: Residents of the Worcester, Mass, area who were hospitalized with non-Q-wave acute myocardial infarction (AMI) at all area hospitals in 5 annual periods between 1990 and 1997 comprised the sample of interest (n = 2436). We examined the extent of use of an invasive versus a conservative strategy for hospital management, occurrence of clinical complications, and hospital mortality in patients with non-Q-wave AMI. RESULTS: An invasive approach to hospital management was used in only 30% of patients with non-Q-wave AMI, although there was an increase over time in the use of an invasive treatment strategy (18% in 1990 vs 33% in 1997). Factors associated with use of the invasive strategy were younger age, male sex, development of cardiogenic shock, as well as adjunctive medical treatment. Overall hospital mortality was high at 12%. Use of the invasive approach, aspirin, nitrates and beta-blockers were associated in logistic regression modeling with significant improvements in survival. CONCLUSIONS: An invasive approach to hospital management of non-Q-wave AMI was used infrequently in this population-based cohort during the 1990s. PMID- 12075261 TI - Prognostic significance of creatine kinase-MB elevation after percutaneous coronary intervention in patients with chronic renal dysfunction. AB - BACKGROUND: Multiple studies have demonstrated a relationship between creatine kinase-MB (CK-MB) elevation after percutaneous coronary intervention (PCI) and increased late mortality within the general population. Because CK-MB is frequently elevated in renal disease even in the absence of myocardial injury, the clinical significance of CK-MB elevation after PCI in patients with renal insufficiency has been questioned. METHODS: We sought to examine the association between elevated CK-MB after PCI and late mortality in 190 consecutive patients with chronic renal insufficiency (serum creatinine > or =2.0 mg/dL) undergoing PCI at the Cleveland Clinic between January 1997 and March 2000. Of the total group, 20 patients undergoing PCI for acute myocardial infarction, cardiogenic shock, or both were excluded. Follow-up was 99.4% complete at a mean duration of 24.8 +/- 11.2 months (range 5-43 months). RESULTS: CK-MB elevation above the upper limit of normal after intervention was detected in 33 patients (19.4%). Baseline characteristics were not significantly different between the CK-MB elevation group and the normal CK-MB group. Late mortality, however, was significantly higher among patients with postprocedural CK-MB elevation (36.4% vs 17.5%, P =.017). Cox proportional hazard model revealed CK-MB elevation as an independent predictor of late mortality (hazard ratio 2.44, 95% CI 1.14-5.24, P =.02), in addition to New York Heart Association class (hazard ratio 1.35, 95% CI 1.05-1.73, P =.02). CONCLUSIONS: This analysis of patients with chronic renal insufficiency undergoing PCI suggests that postprocedural CK-MB elevation is an independent predictor of late mortality even in the presence of renal dysfunction. PMID- 12075262 TI - Magnetic resonance tagging and echocardiographic response to dobutamine and functional improvement after reperfused myocardial infarction. AB - OBJECTIVE: Our objective was to compare the qualitative response to low-dose dobutamine by echocardiography (DSE) with the quantitative response of magnetic resonance myocardial tagging (DMRT) in the prediction and evaluation of functional improvement after reperfused myocardial infarction (MI). METHODS: Twenty-two patients with a reperfused first MI (aged 51 +/- 2 years, 20 male, 13 anterior MI) were studied. On day 3 +/- 1 after MI, patients underwent both DSE and DMRT at baseline and during infusion of 5 microg/kg/min and 10 microg/kg/min of dobutamine. The patients returned at week 8 +/- 1 for follow-up echocardiogram and MRT at rest. Two experienced observers interpreted the DSE for the presence of contractile reserve and functional improvement in dysfunctional segments. By DMRT, a 5% increase in percent intramyocardial circumferential shortening at peak response to dobutamine was defined as evidence of contractile reserve. Functional improvement by echocardiography was defined as the gold standard. RESULTS: Ejection fraction improved from 46% +/- 10% at week 1 to 51% +/- 12% at week 8 (P <.001) in the patients. Sixty-seven transmural segments with baseline dysfunction matched between imaging modalities by location were studied. For 51 (76%) of the segments, echocardiography and MR tagging were concordant in the assessment of functional improvement (kappa value 0.52). Twenty-nine segments (43%) demonstrated improvement by echocardiography, whereas 33 segments (49%) improved by MR tagging. With improvement of function by echocardiography as gold standard, the sensitivity and specificity of DMRT for prediction of functional improvement was 86% and 69%, respectively, with an overall accuracy of 76%. The sensitivity, specificity, and accuracy of DSE was 86%, 87%, and 85%, respectively. Overall accuracy was similar between techniques. CONCLUSIONS: Both DSMRT and DSE are sensitive and accurate techniques for predicting functional improvement after reperfused MI. PMID- 12075263 TI - Do transmyocardial and percutaneous laser revascularization induce silent ischemia? An assessment by exercise testing. AB - BACKGROUND: Transmyocardial and percutaneous laser revascularization (TMR, PTMR) may reduce angina and increase exercise tolerance in otherwise untreatable angina patients, although the mechanism is unknown and the placebo effect may be significant. One other proposed mechanism is cardiac denervation leading to silent ischemia. METHODS: Electrocardiograms obtained during symptom-limited exercise (ETT, modified Bruce protocol) at baseline and 12 months were analyzed (blinded core laboratory) from 182 patients randomized to TMR (n = 92) or medical therapy alone (MED(TMR), n = 90) and 219 patients randomized to PTMR (n = 109) or medical therapy alone (MED(PTMR), n = 110). RESULTS: Exercise duration increased 1 year after TMR or PTMR relative to medically treated patients (6.8 +/- 3.4 min vs 8.6 +/- 3.5 min for TMR; 7.3 +/- 3.1 min vs 9.1 +/- 3.6 min for PTMR, P <.05). At baseline, 20% of TMR and MED(TMR) subjects had ST depression >1.0 mm, >80% had angina during exercise, but only 3% had ST changes without chest pain (silent ischemia). This did not change after TMR. In the PTMR group, more subjects exercised to >1.0 mm ST depression (from 17% to 34%, P <.05), with no change in MED(PTMR), but the proportion with silent ischemia did not change in either group. CONCLUSION: Exercise tolerance improved after TMR and after PTMR. Relative to PTMR, TMR more effectively suppressed pain during exercise and ischemic ST depression. However, neither TMR nor PTMR induced significant silent ischemia. These results suggest that denervation may not be a significant factor contributing to angina relief after these procedures. The contribution of the placebo effect was not determined by these results. PMID- 12075264 TI - Differences between African Americans and whites in the outcome of heart failure: Evidence for a greater functional decline in African Americans. AB - BACKGROUND: National statistics indicate that African Americans are disproportionately affected by mortality and hospitalizations resulting from heart failure when compared with other racial/ethnic groups. This might, in part, reflect a poorer course of heart failure among African Americans. METHODS: We conducted a prospective cohort study of 316 white and 82 African American consecutive patients aged > or =50 years with decompensated heart failure on hospital admission. The outcome of the study was death or decline in activities of daily living function at 6 months relative to baseline. RESULTS: African American patients were on average 8 years younger and had less favorable socioeconomic and access-to-care indicators. African Americans more often had a history of hypertension, renal insufficiency, and diabetes, but there were no differences in functional status, self-reported health status, signs of decompensation, or left ventricular ejection fraction. Quality-of-care indicators did not differ by race. Mortality rates at 6 months were similar in African Americans and whites (19.5% vs 17.2%, age adjusted), but African Americans had a greater functional decline (37.6% vs 24.7%). After adjusting for baseline characteristics, African Americans had an almost 50% higher risk of either death or decline in activities of daily living functioning (relative risk 1.45, 95% CI, 1.06-1.81). Adjustment for socioeconomic, access-to-care and quality-of-care indicators did not substantially change this estimate. CONCLUSIONS: African Americans have similar mortality but greater functional decline than whites after hospitalization for heart failure. This outcome is not explained by clinical, socioeconomic, access-to-care or quality-of-care differences. PMID- 12075265 TI - Evaluation of platelets in heart failure: is platelet activity related to etiology, functional class, or clinical outcomes? AB - OBJECTIVES: We sought to determine whether platelet activity in patients with heart failure is related to an ischemic versus nonischemic etiologic condition, clinical disease severity, or adverse clinical outcomes. BACKGROUND: Platelet activity may affect outcome in patients with heart failure. A prospective evaluation of the relation of baseline platelet function to etiologic condition, New York Heart Association (NYHA) class, and clinical outcomes has not been previously reported. METHODS: Ninety-six consecutive outpatients with ambulatory heart failure with an ejection fraction <0.40 and NYHA Class II to IV symptoms who presented to the Duke Heart Failure Clinic and 14 healthy control subjects formed the study groups. Baseline characteristics and blood analyzed for thromboxane (Tx) B2, 6-keto PGF(1alpha), platelet contractile force, adenosine diphosphate/collagen shear-induced closure time, whole blood aggregation and CD41, CD31, CD62p, and CD51/CD61 by flow cytometry were determined. Survival status and hospitalizations were determined in the heart failure patient cohort. RESULTS: The median age of patients was 65 years (22% female, 64% white). An ischemic etiologic condition was present in 61% of patients. The population had mild to moderate heart failure: NYHA class I (1%), II (41%), III (46%), and IV (12.5%) and severe ventricular dysfunction (median ejection fraction = 0.20). There were 39 clinical events (7 deaths, 3 cardiac transplants, 29 other first hospitalizations) in 305 median days of observation. Platelet activity, indicated by whole blood aggregation with 5 micromol adenosine diphosphate (P =.04) and Tx B2 (P =.01), was higher in patients with heart failure. Whole blood aggregation was greater than the 90th percentile in 22% of patients with heart failure versus 7% of control subjects. Platelet function did not differ for any of the markers between the ischemic and nonischemic groups and was not affected by antecedent aspirin. There was no relation of NYHA class or the occurrence of events to platelet activity. CONCLUSION: Platelet activity is heightened in 22% of outpatients with stable heart failure symptoms and is not affected by antecedent aspirin therapy. The degree of platelet activation is similar in ischemic and nonischemic patients with heart failure and is not related to clinical disease severity. Current methods to assess platelet activation do not appear to predict outcome. PMID- 12075266 TI - Immunoglobulin G3 cardiac myosin autoantibodies correlate with left ventricular dysfunction in patients with dilated cardiomyopathy: immunoglobulin G3 and clinical correlates. AB - BACKGROUND: Effector functions of an aberrant immune response have been implicated in the pathogenesis of idiopathic dilated cardiomyopathy (DCM). The immunologic determinants of myocardial dysfunction, however, remain poorly understood. This study sought to determine the relation of different immunologic responses to hemodynamic dysfunction in DCM. METHODS: Immunoglobulin (Ig) G class/subclass response ELISA (enzyme-linked immunosorbent assay) against cardiac myosin heavy chain, histologic characteristics (DALLAS criteria), immunohistochemistry, plasma interleukin-4 and plasma interferon gamma (IFN gamma) were determined in patients (n = 76) with clinically suspected myocarditis or DCM. Patients were prospectively evaluated, both clinically and hemodynamically, on admission (baseline) and at 6-month follow-up. RESULTS: Indices of hemodynamic dysfunction (by cardiac catheterization and transthoracic echocardiography) correlated significantly with an Ig subclass response. IgG3 levels correlated with left ventricular ejection fraction (P =.02), pulmonary capillary wedge pressure (P <.0001), left ventricular end-systolic volume index (P =.002), left ventricular end-diastolic volume index (P =.033), left ventricular end-diastolic pressure (P =.04), right ventricular end-diastolic pressure (P =.039), and left ventricular end-systolic dimension and left ventricular end-diastolic dimension (P <.05). Patients positive for IgG3 (predominantly male, P =.01) had depressed left ventricular ejection fraction (< or =45%, relative risk 3.0, 95% CI 1.5-5.7, P =.005) at baseline and 6 months. Mitral-septal separation at follow-up improved in patients negative for IgG3 (P =.018), and the number of patients on conventional therapy in this group declined at 6-month follow-up (P <.05). Lymphocyte counts/high-power field; CD2, CD3, CD4, and CD8 (independent of IgG class/subclass response and left ventricular dysfunction) were significantly higher in patients positive for IFN-gamma (25%). A positive IFN-gamma response was higher in patients positive for IgG3. These patients, positive for IgG3 and IFN-gamma (10%), had significantly shorter duration of clinical symptoms: 0.17 years (0.12-2.36 y) versus 1.01 years (0.49 5.35 y, P =.04). CONCLUSION: IgG3 reactivity correlated with depressed myocardial dysfunction. This may render this subclass Ig a surrogate target for therapeutic intervention in DCM. With IFN-gamma, IgG3 may reflect a more aggressive disease. PMID- 12075267 TI - QRS duration and mortality in patients with congestive heart failure. AB - BACKGROUND AND OBJECTIVES: It has been suggested that prolongation of the QRS duration (>120 ms) is an independent risk factor for mortality in patients with cardiomyopathy. The purpose of this study was to examine the association between QRS duration and survival in patients with heart failure. METHODS: We performed a retrospective analysis to examine the association between QRS prolongation (> or =120 ms) and mortality. The study population included 669 patients with heart failure. Two groups, on the basis of baseline QRS duration <120 milliseconds or > or =120 milliseconds, were identified. The groups were compared with respect to total mortality and sudden death. Subgroups were also stratified by right bundle branch block and left bundle branch block, ejection fraction (EF) <30% and > or =30% to 40%, ischemic and nonischemic cardiomyopathy, amiodarone and placebo. RESULTS: Prolonged QRS was associated with a significant increase in mortality (49.3% vs 34.0%, P =.0001) and sudden death (24.8% vs 17.4%, P =.0004). Left bundle branch block was associated with worse survival (P =.006) but not sudden death. In patients with an EF <30%, QRS prolongation continued to be associated with a significant increase in mortality (51.6% vs 41.1%, P =.01) and sudden death (28.8% vs 21.1%, P =.02). In those with an EF of 30% to 40%, QRS prolongation was associated with a significant increase in mortality (42.7% vs 23.3%, P =.0036) but not in sudden death (13.3% vs 12.0%, P =.625). After adjustment for baseline variables, independent predictors of mortality were found to be prolongation of QRS (P =.0028, risk ratio 1.46) and depressed EF (P =.0001, risk ratio 0.965). Age, type of cardiomyopathy, and drug treatment group were not predictive of mortality. CONCLUSION: QRS prolongation is an independent predictor of both increased total mortality and sudden death in patients with heart failure. PMID- 12075268 TI - Nutritional supplementation with MyoVive repletes essential cardiac myocyte nutrients and reduces left ventricular size in patients with left ventricular dysfunction. AB - BACKGROUND: Congestive heart failure depletes the myocardium of carnitine, coenzyme Q10 (CoQ10), and taurine--substances known to influence mitochondrial function and cell calcium. We hypothesized that feeding patients a nutritional supplement that contained carnitine, CoQ10, and taurine would result in higher myocardial levels of these nutrients and improve left ventricular function. METHODS: Forty-one patients who underwent aortocoronary artery bypass with an ejection fraction < or =40% at referral were randomly assigned to a double-blind trial of supplement or placebo. Radionuclide ventriculography was performed at randomization and before surgery. Surgical myocardial biopsies, adjusted for protein content, were analyzed for carnitine, CoQ10, and taurine levels. RESULTS: The groups were well matched. Minor exceptions were supplement group versus placebo group for digoxin use (7 vs 0, respectively; P =.009) and age (62 +/- 11 years vs 69 +/- 5 years, respectively; P =.04). There were significantly higher levels in the treated group compared with the placebo group for myocardial levels of CoQ10 (138.17 +/- 39.87 nmol/g wet weight and 56.67 +/- 23.08 nmol/g wet weight; P =.0006), taurine (13.12 +/- 4.00 micromol/g wet weight and 7.91 +/- 2.81 micromol/g wet weight; P =.003), and carnitine (1735.4 +/- 798.5 nmol/g wet weight and 1237.6 +/- 343.1 nmol/g wet weight; P =.06). The left ventricular end diastolic volume fell by -7.5 +/- 21.7 mL in the supplement group and increased by 10.0 +/- 19.8 mL in the placebo group (P =.037). CONCLUSIONS: Supplementation results in higher myocardial CoQ10, taurine, and carnitine levels and is associated with a reduction in left ventricular end-diastolic volume in patients with left ventricular dysfunction before revascularization. Because the risk of death for surgical revascularization is related to preoperative left ventricular end-diastolic volume, supplementation could improve outcomes. PMID- 12075269 TI - Deceleration time of early filling in patients with left ventricular systolic dysfunction: functional and prognostic independent value. AB - BACKGROUND: Although diastolic function parameters have been mentioned as significant predictors of functional capacity and prognosis in patients with left ventricular (LV) systolic dysfunction, it has not been fully elucidated whether they keep an independent predictive value when multiple parameters from a wide variety of examinations are considered. METHODS: We prospectively studied 60 patients with New York Heart Association (NYHA) class II-IV chronic heart failure symptoms and LV ejection fraction <0.4. At the time of entry into the study, demographic data and functional class were obtained, and usual Doppler echocardiographic, radionuclide ventriculographic, cardiopulmonary exercise testing and hemodynamic variables were determined. Deceleration time of early filling (DT) and NYHA functional class were the only independent predictors of functional capacity as assessed by means of peak oxygen uptake (peak VO2). Mean follow-up was 21 +/- 6 months, and event-free survival was defined as the absence of cardiac death, urgent cardiac transplantation, or hospital admission requiring inotropic or mechanical support. RESULTS: Multivariate Cox analysis showed that DT (P =.008), peak VO2 (P =.01), and NYHA class (P =.02) were independent predictors of event-free survival at 1 year. Patients in the lowest tertile of DT (<130 ms) had a significantly lower event-free survival than patients in the intermediate (44% vs 80%, P =.03) and in the highest tertile (44% vs 83%, P =.02). Patients with both a DT <130 milliseconds and a peak VO2 <14 mL/kg/min had the highest rate of events at 1 year (83% vs 22% for the remaining patients, relative risk 3.75, P <.001). CONCLUSIONS: In patients with LV systolic dysfunction, DT is a powerful independent predictor of functional capacity and prognosis among a wide variety of variables. A shortened DT (<130 ms) identifies a subgroup of patients with a worse outcome, especially when combined with a reduced peak VO2 (<14 mL/kg/min). PMID- 12075270 TI - Elevated serum uric acid levels are associated with diastolic dysfunction in patients with dilated cardiomyopathy. AB - OBJECTIVE: To assess whether serum uric acid, which is a marker of impaired oxidative metabolism, might correlate with left ventricular systolic and diastolic dysfunction in patients with chronic heart failure (CHF). BACKGROUND: Uric acid levels, which are frequently elevated in patients with CHF, correlate with leg vascular resistance. The effects of elevated levels of uric acid on cardiac function in patients with CHF have never been evaluated. METHODS: We studied 150 outpatients with CHF who came to our heart failure clinic. Patients underwent a complete echo-Doppler examination, with measurement of mitral E wave and mitral A wave velocities, E/A ratio, E wave deceleration time (DtE), left ventricular volumes, ejection fraction, and stroke volume. A restrictive mitral filling pattern (RMFP) was defined as either E/A ratio >2 or E/A >1 and DtE <140 milliseconds. RESULTS: Mean age was 62.2 +/- 7.8 years (86% male); 24 patients (16%) had an RMFP. Patients with an RMFP had significantly higher uric acid levels compared with patients without RMFP (0.48 +/- 0.14 mmol/L vs 0.38 +/- 0.08 mmol/L, respectively, P <.001). Uric acid levels correlated significantly with mitral E wave velocity (r =.22, P <.01), E/A ratio (r =.21, P <.05), DtE (r =.26, P <.01), and RMFP (P =.0001). There was no correlation between uric acid and left ventricular volumes, ejection fraction, or stroke volume. In a multivariate model, uric acid predicted DtE independently of renal function, diuretic dose, and left ventricular volumes. CONCLUSION: Elevated uric acid levels are associated with diastolic dysfunction in CHF. Xanthine oxydase inhibition in patients with CHF might theoretically result in an improvement of diastolic function. PMID- 12075271 TI - Vasopeptidase inhibitor reduces inhospital costs for patients with congestive heart failure: results from the IMPRESS trial. Inhibition of Metallo Protease by BMS-186716 in a Randomized Exercise and Symptoms Study in Subjects With Heart Failure. AB - BACKGROUND: The Inhibition of Metallo Protease by BMS-186716 in a Randomized Exercise and Symptoms Study in Subjects With Heart Failure (IMPRESS) clinical trial randomized patients with congestive heart failure to a daily regimen of either omapatrilat or lisinopril. At 24 weeks, patients randomized to omapatrilat had a significant reduction in the combined end point of death, hospitalization, or discontinuation of study drug for worsening heart failure when compared with patients randomized to lisinopril. They also had significantly fewer serious cardiac adverse events. OBJECTIVE: This study sought to determine the economic consequences of the lower event rates of patients who were given omapatrilat. METHODS: Economic outcomes (major hospitalizations and their associated medical costs) were compared between treatment groups and assessed by use of the societal perspective. Hospitalization information was obtained from the IMPRESS trial's standardized case report and serious adverse event forms. Hospital costs were evaluated by means of assigning each hospital admission a diagnosis-related group and an average cost for physician and hospital services. Emergency department visits were included only when they were made for worsening heart failure and were assigned costs equivalent to the average hospital and physician Medicare reimbursement for these visits in Duke University Medical Center's heart failure program. Drug costs were not assessed. RESULTS: Although the typical patient in both treatment groups was event-free, there was a trend toward a greater number of hospitalizations in the patients given lisinopril than in the patients given omapatrilat (P =.07). Differences in the distribution of cardiac hospitalizations between patients given lisinopril and patients given omapatrilat were significant (P =.03). There was a trend toward lower medical costs at 24 weeks in patients given omapatrilat versus patients given lisinopril ($1930 vs $2002, P =.09). Considering only cardiac medical costs, this trend toward reduced medical costs was significant ($1240 vs $1442, P =.03). CONCLUSIONS: At 24 weeks, patients with heart failure treated with omapatrilat had fewer hospitalizations and lower medical costs than patients treated with lisinopril. However, drug treatment costs were not available for this analysis. PMID- 12075272 TI - Prevention of postcoronary angioplasty restenosis by omega-3 fatty acids: main results of the Esapent for Prevention of Restenosis ITalian Study (ESPRIT). AB - BACKGROUND AND OBJECTIVE: Previous trials of omega-3 fatty acids (omega-3 FA) for restenosis prevention after percutaneous transluminal coronary angioplasty (PTCA) have yielded conflicting results. We tested the hypothesis that long-term administration of omega-3 FA before PTCA may have significant effects on restenosis. METHODS: We randomized 339 patients in a double-blind, placebo controlled study of omega-3 FA (as an ethyl ester preparation given as 6 1-g capsules providing 3 g eicosapentaenoic acid and 2.1 g docosahexaenoic acid/d started 1 month before PTCA and given for 1 month thereafter, then continued at half-dose for 6 months) versus an olive oil placebo. Of these, 257 patients (125 on omega-3 FA, 132 on placebo) well matched for risk factors underwent successful balloon-only PTCA (280 total lesions) and were evaluable at 6 months with repeat angiography. Restenosis was defined at quantitative angiography as a recurrence of >50% diameter stenosis in the dilated vessel (Definition I) and as >50% loss of the short-term gain immediately after PTCA (Definition II). RESULTS: Restenosis rates per vessel were 29.4% and 31.6% in the omega-3 FA group, and 39.6% and 35.4% in the placebo group according to Definitions I (P =.04) and II (P = not significant), respectively. Restenosis rates per patient were 31.2% and 33.6% in the omega-3 FA group, and 40.9% and 37.1% in the placebo group according to Definitions I (P =.05) and II (P = not significant), respectively. CONCLUSIONS: With a long treatment before PTCA, omega-3 FA produced a small but significant decrease in the restenosis rate compared with placebo. PMID- 12075275 TI - Nurses Improving Care for Healthsystem Elders (NICHE): using outcomes and benchmarks for evidenced-based practice. AB - This article describes Nurses Improving Care for Healthsystem Elders (NICHE), a project begun in 1992 with four pilot hospitals. These pilot hospitals gathered baseline data using a geriatric institutional assessment profile (GIAP) with a pre- and postdesign to capture changes in staff attitudes, knowledge, and perceptions of the care of older adults. Based on the success of the pilot effort, NICHE, now in its eighth year, has evolved into a program that involves 32 health systems comprising 105 hospitals nationally. To date, more than 10,000 GIAPs have been collected by NICHE hospital staff. All NICHE settings are able to benchmark their GIAP data against comparable institutions (eg, urban, rural, university, community settings) to understand how they compare and then interpret the data at their unique sites. The opportunities for continuous quality improvement through the NICHE program are described. PMID- 12075276 TI - Sustaining the Geriatric Resource Nurse Model at the University of Virginia. AB - This article describes the continued evolution of the Geriatric Resource Nurse Model at the University of Virginia, one of the original NICHE (Nurses Improving Care for Healthsystem Elders) sites established in 1994. The model developed slowly, unit by unit, a few nurses at a time, and has evolved with a multidisciplinary focus. Flexibility, mentoring relationships, a core curriculum, additional stimulating educational offerings, and a strong leadership team have been key influences on sustainability and growth. PMID- 12075277 TI - TWICE: A NICHE program at North Memorial Health Care. AB - Becoming a site for Nurses Improving Care for Healthsystem Elders (NICHE) is an exciting way to improve the care of hospitalized elders. This article describes how a community-based hospital implemented the Geriatric Resource Nurse (GRN) Model on an orthopedic and acute medical surgical unit. Key elements of the program included focusing on a specific geriatric syndrome (acute confusion) and using outcome data to target practice changes. As a result, the incidence of acute confusion and the percentage of acutely confused patients at discharge decreased. PMID- 12075278 TI - The Geriatric Resource Nurse Model: a culture change. AB - The introduction of the Geriatric Resource Nurse (GRN) Model and the Nurses Improving Care for Healthsystem Elders (NICHE) program has changed the culture at Waukesha Memorial Hospital. The number one key to successfully implementing them has been the overwhelming administrative and staff support. The following article contains an outline of the initial steps taken to implement the GRN model and a NICHE program, the way both became fully integrated into the culture of care, and evidence of positive patient and staff outcomes. PMID- 12075279 TI - Best care for the elderly at Forsyth Medical Center. AB - Health care organizations are very interested in the unique needs and resource utilization the aging population will create. This article describes the commitment of a large acute-care hospital to create a senior-friendly environment. The care of hospitalized elders has become a central focus to identify and prioritize improvement opportunities. An interdisciplinary team of professionals passionate about the care of the elder population was developed to champion the efforts, and the NICHE project was their starting point. This article describes the team and the methods they used to modify models and design processes to meet the needs of the organization and this patient population. Successes and opportunities for continued improvement also are identified. PMID- 12075280 TI - Optimal staffing for Acute Care of the Elderly (ACE) units. AB - Acute Care of the Elderly (ACE) units have been shown to improve outcomes for hospitalized elders. Because the literature offers little to describe the appropriate staffing of such units, we surveyed ACE units about their size, configuration, staffing, patient selection, training, and outcomes data. Although we had anticipated speaking with staff from 30 to 40 units, we were able to poll key personnel on only 18 ACE units. This article describes staffing and continuous quality improvement data for these sites. PMID- 12075281 TI - The NICHE LISTSERV. PMID- 12075282 TI - Excelling in advanced practice nursing: Kathleen Ryan Fletcher, RN, APRN, BC, GNP. PMID- 12075283 TI - The relationship of arthritis self-efficacy to functional performance in older men and women with osteoarthritis of the knee. AB - The objective of this study was to examine the relationship between arthritis self-efficacy (ie, the confidence a person has in the ability to perform a specific task) and self-reported functional performance among older men and women with osteoarthritis (OA) of the knee. The findings suggest that the relationship between arthritis self-efficacy and functional performance varies with gender. An older woman's confidence in her ability to perform tasks affects her perceived functional performance. However, this perception may not be true for older men with OA of the knee. PMID- 12075284 TI - Cardiovascular drugs: reason for promise and vigilance. PMID- 12075285 TI - Program of All-inclusive Care for the Elderly in 2002. PMID- 12075286 TI - Treating diabetes with diet. PMID- 12075287 TI - IOLs, innovation, and ethics in pediatric ophthalmology: let's be honest. PMID- 12075288 TI - The value of adult strabismus correction to the patient. AB - OBJECTIVES: To look at the value of adult strabismus correction to the patient, to consider what is known, and to define the challenges and opportunities for helping these patients. METHOD: The benefits of adult strabismus correction leading to the establishment of binocularity, expansion of visual field, and improved psychosocial health are presented and supported by published studies. RESULTS: Adults with strabismus can benefit from its correction. CONCLUSION: There is a tremendous need to educate the medical community and the lay public that adults with strabismus can be helped with restoration of their eyes to a position of alignment for normal function. PMID- 12075289 TI - The long-term torsion effect of the adjustable Harada-Ito procedure. AB - PURPOSE: The Harada-Ito procedure is a widely used technique in the treatment of excyclotorsion. While previous studies have demonstrated its initial success, long-term data have not been readily available. The purpose of this study is to describe the long-term torsion effect of the adjustable Harada-Ito procedure. METHODS: All Harada-Ito procedures performed by 1 surgeon from 1982 to 1998 with at least 1 year of follow-up were reviewed. Torsion was evaluated using the double Maddox rods preoperatively, 1 day postoperatively, 2 months, 6 months, 1 year, 18 months, then annually thereafter. The effectiveness of the procedure was evaluated by comparing preoperative values to those at follow-up exams. The stability of the procedure was also evaluated by comparing torsion at 1 day and 2 months postoperatively to the torsion at subsequent exams. All comparisons were performed using the Student's t test. RESULTS: Eighty Harada-Ito procedures were performed by 1 surgeon during this period. Nineteen patients had at least 1 year of follow-up (range, 1-7 years; mean, 2.0 years). Mean excyclotorsion values were 10.7 degrees preoperatively, 1.0 degrees at 1 day, 1.0 degrees at 2 months, 3.4 degrees at 6 months, 4.8 degrees at 1 year, and 5.3 degrees at 18 months. There was a significant improvement of excyclotorsion compared to preoperative torsion up to 12 months after surgery (P or= 20 degrees) often resulting from macular translocation surgery, but does not allow reestablishment of binocular function. PMID- 12075293 TI - Preoperative prism adaptation for acquired esotropia: long-term results. AB - PURPOSE: The efficacy of preoperative prism adaptation in subjects with acquired esotropia has been established at 6 and 12 months postoperatively. We evaluated the outcomes of subjects with acquired esotropia who had preoperative prism adaptation and were followed longer than 12 months. METHODS: A retrospective analysis was undertaken of 2 groups of subjects with acquired esotropia who underwent bilateral medial rectus recessions based on the distance angle of deviation and were followed more than 12 months postoperatively. Group A subjects had preoperative prism adaptation to determine the target angle for corrective surgery. In Group B subjects, surgery was based on the maximum angle of strabismus at distance, determined by alternate prism cover test without preoperative prism adaptation. RESULTS: We compared 17 subjects in Group A and 19 subjects in Group B. The postoperative follow-up period was 3 +/- 1.7 years in Group A and 4.8 +/- 1.8 years in Group B. The age at the last visit was 9.1 +/- 2 years in Group A and 10 +/- 2 years in Group B. Postoperative residual esotropia was 2.6 +/- 2.5 PD at distance in Group A patients and 6.6 +/- 5.9 PD in group B patients (P =.002). Residual esotropia at near was 3 +/- 3.8 PD in Group A and 11.5 +/- 8.12 PD in Group B (P <.01). More Group B subjects required bifocal spectacles to achieve optimal alignment at near (P =.001). CONCLUSION: Acquired esotropia subjects operated on for their distance prism-adapted angle maintained better motor alignment over a long-term follow-up period when compared with nonprism-adapted subjects operated on for their distance angle. PMID- 12075292 TI - Clinical outcomes of pars plana capsulotomy with anterior vitrectomy in pediatric cataract surgery. AB - PURPOSE: The management of the posterior capsule in pediatric cataract surgery remains challenging. The purpose of our study was to evaluate the role and clinical outcomes of primary pars plana capsulotomy and pars plana anterior vitrectomy during pediatric cataract surgery with intraocular lens (IOL) implantation. METHODS: Consecutive cases of pars plana capsulotomy and anterior vitrectomy during pediatric cataract surgery were retrospectively reviewed. The surgical technique was the same in all patients and involved pars plana capsulotomy with anterior vitrectomy after the IOL was implanted in the capsular bag. RESULTS: Seventy-six pediatric cataract cases in 61 patients performed from 1994 through 1999 used the pars plana posterior capsulotomy technique and were included in this analysis. A posterior chamber IOL was implanted in all cases. There were no intraoperative complications associated with the surgery. Median age at the time of surgery was 21 months. All patients had at least 1 year of follow-up and mean follow-up was 29.4 months. Among patients old enough to read Snellen's letters (53 eyes), final best-corrected visual acuity was 20/40 or better in 32 eyes (60%) and 20/50 to 20/200 in 21 eyes (40%). Poor final visual acuity was associated with deprivation amblyopia or optic nerve dysplasia or hypoplasia. There were no postoperative retinal complications or cases of IOL dislocation during the follow-up period. Seven eyes (9.2%) of five patients with a median age of 2.5 months developed reopacification of the visual axis at a mean of 4.1 months postoperatively. This event occurred in 6 of 28 eyes with an age of 6 months or less (21.4%), versus only 1 of 48 eyes with an age greater than 6 months (2.0%) (P =.006, Fisher exact test). The visual axis was restored by pars plana membranectomy in 6 eyes and 1 eye of one patient underwent neodymium:YAG laser membrane discission. CONCLUSIONS: Pars plana capsulotomy with pars plana anterior vitrectomy is a safe, effective method of managing the posterior capsule in pediatric cataract surgery with IOL implantation. Visual axis reopacification is associated with a very young age at the time of surgery. Continued follow-up of these patients is important to assess the long-term outcomes of this surgical approach. PMID- 12075294 TI - Risk factors for the development of accommodative esotropia following treatment for infantile esotropia. AB - PURPOSE: One aim of the study was to determine whether accommodative esotropia after surgical alignment in infantile esotropia occurs because a pre-existing accommodative component is unmasked at the time of surgery or whether it occurs as a sequela of infantile esotropia. A second aim of the present study was to examine risk factors for accommodative esotropia after surgery for infantile esotropia. METHODS: A total of 80 consecutive patients who were enrolled in a prospective study of infantile esotropia had been followed for more than 4 years and had achieved orthoposition were included. Twelve potential risk factors were examined: age at onset, initial esodeviation, initial refractive error, age at alignment, delay in alignment, presurgical glasses, amblyopia, additional surgical procedures, unstable alignment, increase in hypermetropia, peripheral fusion, and stereopsis. Mantel-Haenszel odds ratios were computed for each factor and were corrected to relative risks. RESULTS: Overall, 48 of 80 children (60%) developed accommodative esotropia at a mean age of 33 months. Increasing hypermetropia, delay in alignment, and poor stereopsis posed significant risks for accommodative esotropia. The remaining 9 factors were not associated with increased risk for accommodative esotropia. CONCLUSIONS: Accommodative esotropia is unlikely to be a pre-existing condition in most cases because the mean age of onset was 23 months postoperative and the prevalence of preoperative hypermetropia greater than +3.00 D was low. Both delay in alignment and stereopsis risk factors may reflect compromised binocular sensory status that allows accommodative esotropia to occur at low to moderate levels of hypermetropia. Identification of children treated for infantile esotropia who are at risk for accommodative esotropia may allow for prevention or early treatment. PMID- 12075295 TI - Results of the modified trabeculotomy in the treatment of primary congenital glaucoma. AB - PURPOSE: To evaluate the effect of trabeculotomy employing probes with curvatures more closely corresponding to variable course of Schlemm's canal. METHODS: Forty six children with primary congenital glaucoma who underwent trabeculotomy with newly designed probes between 1990 and 1998 were evaluated retrospectively. One set of the newly designed instruments consists of 3 pairs of trabeculotomy probes, the curvatures of which correspond to corneal diameters of 10, 12, and 14 mm. The criteria for surgical success were intraocular pressure (IOP) below 21 mmHg and simultaneous absence of cup to disc (C/D) ratio progression and of disproportional enlargement of the cornea. RESULTS: Forty-six children who underwent surgery over the course of 7 years were evaluated in a follow-up period from 7 to 83 months (38.4 +/- 22.5 months). A total of 102 trabeculotomies were performed in a total of 78 eyes, with the following complications: 8 large intraoperative hemorrhages, 3 small iris prolapses, 2 small descemetolyses, and 1 short iridodialysis. Postoperatively, glaucoma was successfully controlled in 68 eyes (87%), IOP fell in average from 30.81 +/- 6.55 mmHg to 16.32 +/- 5.13 mmHg (P <.00005). Visual acuity was better than 0.4 in 50 eyes and worse than 0.1 in 5 eyes at the end of the study. CONCLUSION: The probing of the Schlemm's canal was feasible in all procedures and the number of complications was low. The success rate of surgery was high. Adaptation of the trabeculotomy probes to the Schlemm's canal curvature seems to make probing easier and safer. PMID- 12075296 TI - Evaluation of the ultrasound biomicroscope in strabismus surgery. AB - PURPOSE: This study was done to determine the ability of the ultrasound biomicroscope (UBM) to define the distance of the horizontal extraocular muscle insertions from the limbus in children undergoing strabismus surgery. METHODS: A total of 51 children undergoing primary surgery on their horizontal muscles were recruited for this study. The 50 MHz UBM was used before surgery on the muscle(s) requiring an operation to define the distance of the insertion of the horizontal rectus muscle(s) from the limbus. These data were stored on a computer and masked from the surgeon who measured the actual insertion from the limbus with calipers intraoperatively. An intraclass correlation coefficient was calculated to determine the correlation of the UBM measurement with that performed during surgery. RESULTS: Measurement of the distance of the insertions from the limbus were taken on 79 muscles of the 51 patients: 44 medial rectus (MR) and 35 lateral rectus (LR). Mean age of the patients was 6.4 +/- 3.7 years (range, 1-15). A total of 29 children (56.9%) were female. The mean distance of MR from the limbus measured surgically was 5.2 +/- 0.6 mm (range, 4-6 mm; median, 5 mm) and measured with the UBM was 5.2 +/- 0.9 mm (range, 4-7.6 mm; median, 5.3 mm). Comparable results for the LR muscles were 6.4 +/- 0.6 mm (range, 5.5-7.5 mm; median, 6.5 mm) and 6.9 +/- 0.7 mm (range, 5.9-8.4 mm; median, 6.9 mm), respectively. An intraclass correlation coefficient of 0.71 signified good agreement between the surgical and UBM measurements of the distance from the limbus of the horizontal rectus muscle insertions. CONCLUSION: The UBM could be used reliably as a tool before surgery to estimate the distance of the horizontal rectus muscles from the limbus in children with strabismus. PMID- 12075297 TI - The reliability of grading the fixation preference test for the assessment of interocular visual acuity differences in patients with strabismus. AB - PURPOSE: The purpose of this study is to test the reliability of the grading system of standard fixation preference testing compared with the logarithmic scale of the minimum angle of resolution (logMAR) interocular visual acuity difference of patients with manifest strabismus and to test the strength of association between the interocular visual acuity difference and the grade of alternation. METHODS: A total of 111 strabismic patients with large angle heterotropias (> 10 PD) were included in this study. The visual acuities of these patients were determined using the Snellen's, tumbling E, or Lea visual acuity charts as appropriate for the child's age and ability to cooperate. The results were converted to their logarithmic equivalents of minimum angle of resolution or recognition. Fixation preference was graded from 0 to 4 and was evaluated by observing the time-period through which fixation was maintained with the nonpreferred eye. RESULTS: The median value of interocular visual acuity difference for grade 4 was determined as 0.0, grade 3 as 0.097, grade 2 as 0.17, grade 1 as 0.3, and grade 0 as 0.8. The grade of fixation preference was found to be inversely related to the logMAR interocular visual acuity difference (P <.0001). There was a significant difference between every fixation grade except grade 1 and grade 2 (P <.005). CONCLUSIONS: The grades used in fixation preference testing correlate with the logMAR visual acuity differences in strabismic amblyopic patients. However, the grading system does not imply a linear function in terms of visual acuity difference. The difference between the logMAR values of patients with grade 0 and 1 are more profound compared with other grades. PMID- 12075298 TI - Functional visual loss in patients with bilateral ocular coloboma. PMID- 12075299 TI - Asymmetric "hang-back" superior oblique recession. PMID- 12075301 TI - Equal opportunities. PMID- 12075302 TI - Rethink anti-bioterrorism plans. PMID- 12075303 TI - US urged to provide smallpox vaccines for emergency crews. PMID- 12075304 TI - Senate adrift on cloning as talks break down. PMID- 12075305 TI - 'Dirty bomb' scare prompts clampdown on lab security. PMID- 12075306 TI - Pentagon gets defensive over missile secrets. PMID- 12075307 TI - Astronomers give virtual observatory a real future. PMID- 12075308 TI - Drug researcher quits controversial cat study. PMID- 12075309 TI - Draft cow genome heads the field. PMID- 12075310 TI - Patent flurry casts cloud over gene silencing. PMID- 12075312 TI - Squaring up over ancient life. PMID- 12075313 TI - Full house. PMID- 12075314 TI - Learning from past mistakes about nuclear waste. PMID- 12075315 TI - Macroecology: new, or biogeography revisited? PMID- 12075316 TI - Online database could end taxonomic anarchy. PMID- 12075317 TI - To sell science, find out what people want to buy. PMID- 12075318 TI - Health supercourse to end Arab isolation. PMID- 12075319 TI - Science's policy on access to private data. PMID- 12075321 TI - Is a bell tolling for Bell Labs? PMID- 12075328 TI - Strength in numbers. PMID- 12075329 TI - Teamed up for transcription. PMID- 12075330 TI - Nebulous explanation. PMID- 12075333 TI - Imprints offer Moore. PMID- 12075331 TI - Two ACEs and a heart. PMID- 12075334 TI - Neurobiology: understanding the consequences. PMID- 12075336 TI - Beyond the diffraction limit? PMID- 12075337 TI - Structural biology: a molecular propeller. PMID- 12075339 TI - Only male fin whales sing loud songs. AB - The low-frequency vocalizations of fin and blue whales are the most powerful and ubiquitous biological sounds in the ocean. Here we combine acoustic localization and molecular techniques to show that, in fin whales, only males produce these vocalizations. This finding indicates that they may function as male breeding displays, and will help to focus concern on the impact of human-generated low frequency sounds on recovering whale populations. PMID- 12075340 TI - Physiology: Dynamic instabilities in the inflating lung. AB - In lung diseases such as asthma, expiratory flow becomes limited, airways can collapse and the vital exchange of gases is compromised. Here we model the inflation of collapsed regions of the lung during inspiration in terms of avalanches propagating through a bifurcating network of airways, and find that the accompanying cascade of dynamic pressure instabilities -- avalanche 'shocks' - manifests as negative elastic resistance of the lung. Our analysis of this apparent thermodynamic paradox provides a better understanding of aeration in the deep regions of the lung, which may find application in medical conditions in which gas exchange is impaired. PMID- 12075341 TI - Contact time of a bouncing drop. AB - When a liquid drop lands on a solid surface without wetting it, it bounces with remarkable elasticity. Here we measure how long the drop remains in contact with the solid during the shock, a problem that was considered by Hertz for a bouncing ball. Our findings could help to quantify the efficiency of water-repellent surfaces (super-hydrophobic solids) and to improve water-cooling of hot solids, which is limited by the rebounding of drops as well as by temperature effects. PMID- 12075342 TI - Evolutionary biology: hedgehog crosses the snail's midline. AB - According to the dorsoventral axis-inversion theory, protostomes (such as insects, snails and worms) are organized upside-down by comparison with deuterostomes (vertebrates), in which case their respective ventrally (belly side) and dorsally (back-side) located nervous systems, as well as their midline regions, should all be derived from a common ancestor. Here we provide experimental evidence for such homology by showing that an orthologue of hedgehog, an important gene in midline patterning in vertebrates, is expressed along the belly of the larva of the limpet Patella vulgata. This finding supports the existence of a similar mechanism for the development of the midline of the nervous system in protostomes and deuterostomes. PMID- 12075343 TI - Ordered porous materials for emerging applications. AB - "Space--the final frontier." This preamble to a well-known television series captures the challenge encountered not only in space travel adventures, but also in the field of porous materials, which aims to control the size, shape and uniformity of the porous space and the atoms and molecules that define it. The past decade has seen significant advances in the ability to fabricate new porous solids with ordered structures from a wide range of different materials. This has resulted in materials with unusual properties and broadened their application range beyond the traditional use as catalysts and adsorbents. In fact, porous materials now seem set to contribute to developments in areas ranging from microelectronics to medical diagnosis. PMID- 12075344 TI - Angiotensin-converting enzyme 2 is an essential regulator of heart function. AB - Cardiovascular diseases are predicted to be the most common cause of death worldwide by 2020. Here we show that angiotensin-converting enzyme 2 (ace2) maps to a defined quantitative trait locus (QTL) on the X chromosome in three different rat models of hypertension. In all hypertensive rat strains, ACE2 messenger RNA and protein expression were markedly reduced, suggesting that ace2 is a candidate gene for this QTL. Targeted disruption of ACE2 in mice results in a severe cardiac contractility defect, increased angiotensin II levels, and upregulation of hypoxia-induced genes in the heart. Genetic ablation of ACE on an ACE2 mutant background completely rescues the cardiac phenotype. But disruption of ACER, a Drosophila ACE2 homologue, results in a severe defect of heart morphogenesis. These genetic data for ACE2 show that it is an essential regulator of heart function in vivo. PMID- 12075345 TI - A collimated jet of molecular gas from a star on the asymptotic giant branch. AB - Evolved stars of about one solar mass are in general spherically symmetric, yet the planetary nebulae that they produce in the next phase of their evolution tend not to exhibit such symmetry. Collimated 'jets' and outflows of material have been observed up to approximately 0.3 parsec from the central stars of planetary nebulae, and precession of those jets has been proposed to explain the observed asymmetries. Moreover, it has recently been shown theoretically that magnetic fields could launch and collimate such jets. Here we report the detection of a collimated and precessing jet of molecular gas that is traced by water-vapour maser spots approximately 500 astronomical units (au) from the star W43A in Aquila. We conclude that the jet is formed in the immediate vicinity of the star, and infer that elongated planetary nebulae are formed by jets during the short period, of less than 1,000 years, when the star makes its transition through the proto-planetary nebula phase to become a planetary nebula. PMID- 12075346 TI - Hidden orbital order in the heavy fermion metal URu(2)Si(2). AB - When matter is cooled from high temperatures, collective instabilities develop among its constituent particles that lead to new kinds of order. An anomaly in the specific heat is a classic signature of this phenomenon. Usually the associated order is easily identified, but sometimes its nature remains elusive. The heavy fermion metal URu(2)Si(2) is one such example, where the order responsible for the sharp specific heat anomaly at T(0) = 17 K has remained unidentified despite more than seventeen years of effort. In URu(2)Si(2), the coexistence of large electron electron repulsion and antiferromagnetic fluctuations leads to an almost incompressible heavy electron fluid, where anisotropically paired quasiparticle states are energetically favoured. Here we develop a proposal for the nature of the hidden order in URu(2)Si(2). We show that incommensurate orbital antiferromagnetism, associated with circulating currents between the uranium ions, can account for the local fields and entropy loss observed at the 17 K transition. We make detailed predictions for the outcome of neutron scattering measurements based on this proposal, so that it can be tested experimentally. PMID- 12075347 TI - Ultrafast and direct imprint of nanostructures in silicon. AB - The fabrication of micrometre- and nanometre-scale devices in silicon typically involves lithography and etching. These processes are costly and tend to be either limited in their resolution or slow in their throughput. Recent work has demonstrated the possibility of patterning substrates on the nanometre scale by 'imprinting' or directed self-assembly, although an etching step is still required to generate the final structures. We have devised and here demonstrate a rapid technique for patterning nanostructures in silicon that does not require etching. In our technique which -- we call 'laser-assisted direct imprint' (LADI) -- a single excimer laser pulse melts a thin surface layer of silicon, and a mould is embossed into the resulting liquid layer. A variety of structures with resolution better than 10 nm have been imprinted into silicon using LADI, and the embossing time is less than 250 ns. The high resolution and speed of LADI, which we attribute to molten silicon's low viscosity (one-third that of water), could open up a variety of applications and be extended to other materials and processing techniques. PMID- 12075348 TI - Growth of early continental crust controlled by melting of amphibolite in subduction zones. AB - It is thought that the first continental crust formed by melting of either eclogite or amphibolite, either at subduction zones or on the underside of thick oceanic crust. However, the observed compositions of early crustal rocks and experimental studies have been unable to distinguish between these possibilities. Here we show a clear contrast in trace-element ratios of melts derived from amphibolites and those from eclogites. Partial melting of low-magnesium amphibolite can explain the low niobium/tantalum and high zirconium/samarium ratios in melts, as required for the early continental crust, whereas the melting of eclogite cannot. This indicates that the earliest continental crust formed by melting of amphibolites in subduction-zone environments and not by the melting of eclogite or magnesium-rich amphibolites in the lower part of thick oceanic crust. Moreover, the low niobium/tantalum ratio seen in subduction-zone igneous rocks of all ages is evidence that the melting of rutile-eclogite has never been a volumetrically important process. PMID- 12075349 TI - Paired gill slits in a fossil with a calcite skeleton. AB - The chordates, hemichordates (such as acorn worms) and echinoderms (such as starfish) comprise the group Deuterostomia, well established as monophyletic. Among extant deuterostomes, a skeleton in which each plate has the crystallographic structure of a single crystal of calcite is characteristic of echinoderms and is always associated with radial symmetry and never with gill slits. Among fossils, however, such a skeleton sometimes occurs without radial symmetry. This is true of Jaekelocarpus oklahomensis, from the Upper Carboniferous of Oklahoma, USA, which, being externally almost bilaterally symmetrical, is traditionally placed in the group Mitrata (Ordovician to Carboniferous periods, 530-280 million years ago), by contrast with the bizarrely asymmetrical Cornuta (Cambrian to Ordovician periods, 540 to 440 million years ago). Using computer X-ray microtomography, we describe the anatomy of Jaekelocarpus in greater detail than formerly possible, reveal evidence of paired gill slits internally and interpret its functional anatomy. On this basis we suggest its phylogenetic position within the deuterostomes. PMID- 12075350 TI - Positive interactions among alpine plants increase with stress. AB - Plants can have positive effects on each other. For example, the accumulation of nutrients, provision of shade, amelioration of disturbance, or protection from herbivores by some species can enhance the performance of neighbouring species. Thus the notion that the distributions and abundances of plant species are independent of other species may be inadequate as a theoretical underpinning for understanding species coexistence and diversity. But there have been no large scale experiments designed to examine the generality of positive interactions in plant communities and their importance relative to competition. Here we show that the biomass, growth and reproduction of alpine plant species are higher when other plants are nearby. In an experiment conducted in subalpine and alpine plant communities with 115 species in 11 different mountain ranges, we find that competition generally, but not exclusively, dominates interactions at lower elevations where conditions are less physically stressful. In contrast, at high elevations where abiotic stress is high the interactions among plants are predominantly positive. Furthermore, across all high and low sites positive interactions are more important at sites with low temperatures in the early summer, but competition prevails at warmer sites. PMID- 12075351 TI - Consumer versus resource control of species diversity and ecosystem functioning. AB - A key question in ecology is which factors control species diversity in a community. Two largely separate groups of ecologists have emphasized the importance of productivity or resource supply, and consumers or physical disturbance, respectively. These variables show unimodal relationships with diversity when manipulated in isolation. Recent multivariate models, however, predict that these factors interact, such that the disturbance diversity relationship depends on productivity, and vice versa. We tested these models in marine food webs, using field manipulations of nutrient resources and consumer pressure on rocky shores of contrasting productivity. Here we show that the effects of consumers and nutrients on diversity consistently depend on each other, and that the direction of their effects and peak diversity shift between sites of low and high productivity. Factorial meta-analysis of published experiments confirms these results across widely varying aquatic communities. Furthermore, our experiments demonstrate that these patterns extend to important ecosystem functions such as carbon storage and nitrogen retention. This suggests that human impacts on nutrient supply and food-web structure have strong and interdependent effects on species diversity and ecosystem functioning, and must therefore be managed together. PMID- 12075352 TI - A global analysis of Caenorhabditis elegans operons. AB - The nematode worm Caenorhabditis elegans and its relatives are unique among animals in having operons. Operons are regulated multigene transcription units, in which polycistronic pre-messenger RNA (pre-mRNA coding for multiple peptides) is processed to monocistronic mRNAs. This occurs by 3' end formation and trans splicing using the specialized SL2 small nuclear ribonucleoprotein particle for downstream mRNAs. Previously, the correlation between downstream location in an operon and SL2 trans-splicing has been strong, but anecdotal. Although only 28 operons have been reported, the complete sequence of the C. elegans genome reveals numerous gene clusters. To determine how many of these clusters represent operons, we probed full-genome microarrays for SL2-containing mRNAs. We found significant enrichment for about 1,200 genes, including most of a group of several hundred genes represented by complementary DNAs that contain SL2 sequence. Analysis of their genomic arrangements indicates that >90% are downstream genes, falling in 790 distinct operons. Our evidence indicates that the genome contains at least 1,000 operons, 2 8 genes long, that contain about 15% of all C. elegans genes. Numerous examples of co-transcription of genes encoding functionally related proteins are evident. Inspection of the operon list should reveal previously unknown functional relationships. PMID- 12075353 TI - Feedback inhibition controls spike transfer in hybrid thalamic circuits. AB - Sensory information reaches the cerebral cortex through the thalamus, which differentially relays this input depending on the state of arousal. Such 'gating' involves inhibition of the thalamocortical relay neurons by the reticular nucleus of the thalamus, but the underlying mechanisms are poorly understood. We reconstructed the thalamocortical circuit as an artificial and biological hybrid network in vitro. With visual input simulated as retinal cell activity, we show here that when the gain in the thalamic inhibitory feedback loop is greater than a critical value, the circuit tends towards oscillations -- and thus imposes a temporal decorrelation of retinal cell input and thalamic relay output. This results in the functional disconnection of the cortex from the sensory drive, a feature typical of sleep states. Conversely, low gain in the feedback inhibition and the action of noradrenaline, a known modulator of arousal, converge to increase input output correlation in relay neurons. Combining gain control of feedback inhibition and modulation of membrane excitability thus enables thalamic circuits to finely tune the gating of spike transmission from sensory organs to the cortex. PMID- 12075354 TI - Na(+)/H(+ ) exchanger regulatory factor 2 directs parathyroid hormone 1 receptor signalling. AB - The parathyroid hormone 1 receptor (PTH1R) is a class II G-protein-coupled receptor. PTH1R agonists include both PTH, a hormone that regulates blood calcium and phosphate, and PTH-related protein (PTHrP), a paracrine/autocrine factor that is essential for development, particularly of the skeleton. Adenylyl cyclase activation is thought to be responsible for most cellular responses to PTH and PTHrP, although many actions appear to be independent of adenylyl cyclase. Here we show that the PTH1R binds to Na(+)/H(+) exchanger regulatory factors (NHERF) 1 and 2 through a PDZ-domain interaction in vitro and in PTH target cells. NHERF2 simultaneously binds phospholipase C beta 1 and an atypical, carboxyl-terminal PDZ consensus motif, ETVM, of the PTH1R through PDZ1 and PDZ2, respectively. PTH treatment of cells that express the NHERF2 PTH1R complex markedly activates phospholipase C beta and inhibits adenylyl cyclase through stimulation of inhibitory G proteins (G(i/o) proteins). NHERF-mediated assembly of PTH1R and phospholipase C beta is a unique mechanism to regulate PTH signalling in cells and membranes of polarized cells that express NHERF, which may account for many tissue- and cell-specific actions of PTH/PTHrP and may also be relevant to signalling by many G-protein-coupled receptors. PMID- 12075355 TI - TNF-mediated inflammatory skin disease in mice with epidermis-specific deletion of IKK2. AB - The I kappa B kinase (IKK), consisting of the IKK1 and IKK2 catalytic subunits and the NEMO (also known as IKK gamma) regulatory subunit, phosphorylates I kappa B proteins, targeting them for degradation and thus inducing activation of NF kappa B (reviewed in refs 1, 2). IKK2 and NEMO are necessary for NF-kappa B activation through pro-inflammatory signals. IKK1 seems to be dispensable for this function but controls epidermal differentiation independently of NF-kappa B. Previous studies suggested that NF-kappa B has a function in the growth regulation of epidermal keratinocytes. Mice lacking RelB or I kappa B alpha, as well as both mice and humans with heterozygous NEMO mutations, develop skin lesions. However, the function of NF-kappa B in the epidermis remains unclear. Here we used Cre/loxP-mediated gene targeting to investigate the function of IKK2 specifically in epidermal keratinocytes. IKK2 deficiency inhibits NF-kappa B activation, but does not lead to cell-autonomous hyperproliferation or impaired differentiation of keratinocytes. Mice with epidermis-specific deletion of IKK2 develop a severe inflammatory skin disease, which is caused by a tumour necrosis factor-mediated, alpha beta T-cell-independent inflammatory response that develops in the skin shortly after birth. Our results suggest that the critical function of IKK2-mediated NF-kappa B activity in epidermal keratinocytes is to regulate mechanisms that maintain the immune homeostasis of the skin. PMID- 12075356 TI - Mice deficient in the Rac activator Tiam1 are resistant to Ras-induced skin tumours. AB - Proteins of the Rho family control signalling pathways that regulate the actin cytoskeleton and gene transcription. In vitro studies have implicated Rho-like GTP-hydrolysing enzymes (GTPases) in cell migration, cell-cycle progression, and Ras-induced focus formation, suggesting a role for these GTPases in the formation and progression of tumours in vivo. To study this, we have generated mice lacking the Rac-specific activator Tiam1, a T-lymphoma invasion and metastasis inducing protein. Here we show that such Tiam1(-/-) mice are resistant to the development of Ras-induced skin tumours initiated with 7,12-dimethylbenzanthracene and promoted with 12-O-tetradecanoylphorbol-13-acetate. Moreover, the few tumours produced in Tiam1(-/-) mice grew much slower than did tumours in wild-type mice. Tiam1-deficient primary embryonic fibroblasts were also resistant to Ras(V12) induced focus formation. Analysis of Tiam1 heterozygotes indicated that both tumour initiation and promotion were dependent on the Tiam1 gene dose. Tiam1 deficiency was associated with increased apoptosis during initiation, and with impeded proliferation during promotion. Although the number of tumours in Tiam1( /-) mice was small, a greater proportion progressed to malignancy, suggesting that Tiam1 deficiency promotes malignant conversion. Our studies identify the Rac activator Tiam1 as a critical regulator of different aspects of Ras-induced tumour formation. PMID- 12075357 TI - Efp targets 14-3-3 sigma for proteolysis and promotes breast tumour growth. AB - Oestrogen exerts its influence on target organs through activating oestrogen receptors (ERs) and regulating downstream genes by means of their oestrogen responsive elements. Efp, a target gene product of ER alpha, is a member of the RING-finger B-box coiled-coil (RBCC) motif family. Efp is predominantly expressed in various female organs as well as in breast cancers, and is thought to be essential for oestrogen-dependent cell proliferation and organ development Efp disrupted mice display underdeveloped uteri and reduced oestrogen responsiveness. Here we show that Efp is a RING-finger-dependent ubiquitin ligase (E3) that targets proteolysis of 14-3-3 sigma, a negative cell cycle regulator that causes G2 arrest. We demonstrate that tumour growth of breast cancer MCF7 cells implanted in female athymic mice is reduced by treatment with antisense Efp oligonucleotide. Efp-overexpressing MCF7 cells in ovariectomized athymic mice generate tumours in the absence of oestrogen. Loss of Efp function in mouse embryonic fibroblasts results in an accumulation of 14-3-3 sigma, which is responsible for reduced cell growth. These data provide an insight into the cell cycle machinery and tumorigenesis of breast cancer by identifying 14-3-3 sigma as a target for proteolysis by Efp, leading to cell proliferation. PMID- 12075360 TI - More vaccines? Using economic analysis to decide. PMID- 12075361 TI - Economic analyses of rubella and rubella vaccines: a global review. AB - OBJECTIVE: To investigate whether the incorporation of rubella vaccine into immunization programmes in developing countries is economically justified. METHODS: A MEDLINE search was conducted for articles published between 1970 and 2000 that dealt with economic analyses of rubella and rubella-containing vaccines. The Eastern Mediterranean, South-East Asia, and Africa regional Index Medicus databases and the LILACS database for Latin America and the Caribbean were also searched. FINDINGS: For developed countries, five cost- benefit analyses of rubella vaccine and five of measles-mumps-rubella vaccine as well as two cost-effectiveness analyses were found. For developing countries, five cost analyses and five cost-benefit analyses were found. All the cost-benefit analyses had a benefit:cost ratio greater than 1 and the cost-effectiveness studies indicated that rubella immunization was a cost-effective means of reducing the impact of congenital rubella syndrome. However, the methodologies were not standardized. CONCLUSION: The data support the inclusion of rubella vaccine in the immunization programmes of both developing and developed countries and indicate economic benefits comparable to those associated with hepatitis B vaccine and Haemophilus influenzae type b vaccine. More studies should be carried out on costs for care and immunization using standardized methodologies and locally obtained information. PMID- 12075362 TI - Time to focus child survival programmes on the newborn: assessment of levels and causes of infant mortality in rural Pakistan. AB - OBJECTIVE: Population-based surveys were conducted in selected clusters of Pakistan's least developed provinces, Balochistan and North-West Frontier Province (NWFP), including the Federally Administered Tribal Areas (FATA), to assess levels and causes of neonatal and postneonatal mortality. METHODS: Interviews were conducted in a total of 54 834 households: Balochistan, 20 486; NWFP, 26 175; and FATA, 8173. Trained interviewers administered questionnaires after obtaining verbal informed consent from the respondents. Verbal autopsy interviews were conducted for infant deaths reported for the previous year. FINDINGS: The infant mortality rate based on combined data from the different sites was 99.7 per 1000 live births (range 129.0-70.1). The contribution of neonatal deaths to all infant deaths was much higher for NWFP (67.2%), where the overall rate was lowest, than for Balochistan (50.8%) and FATA (56.8%). Around 70% of all neonatal deaths occurred in the early neonatal period. The three main clinical causes of infant deaths were diarrhoea syndrome (21.6%), tetanus (11.7%) and acute respiratory infections (11.6%). In the neonatal period, however, tetanus (18.3%), small size for gestational age or low birth weight (15.3%), and birth injury (12.0%) accounted for nearly half (45.6%) of all deaths, while the contributions of diarrhoea syndrome (5.1%) and acute respiratory infections (6.0%) were less significant (11.1%). Tetanus was the cause of death for 21.7% and 17.1% of all infant deaths in FATA and NWFP respectively. CONCLUSION: The results suggest that there should be a shift in child survival programmes to give greater emphasis to maternal and neonatal health, in particular to maternal tetanus immunization, safe delivery and cord care. PMID- 12075363 TI - Universal immunization of infants with low doses of a low-cost, plasma-derived hepatitis B vaccine in South Africa. AB - OBJECTIVE: To evaluate the effectiveness of universal vaccination against viral hepatitis B in South Africa among 18-month-old rural children. METHODS: Children were immunized with a course of low-dose (1.5 microg), plasma-derived hepatitis B vaccine at 6, 10 and 14 weeks of age, and blood samples from the children were tested for three hepatitis B markers: hepatitis B surface antigen (HBsAg), anti HBs and anti-HBc. FINDINGS: One year after vaccination, a protective anti-HBs antibody titre of at least 10 IU/l was present in 669/769 (87.0%) of blood serum samples tested. Only 3/756 children (0.4%) were HBsAg positive and a fourth child was anti-HBc positive (HBsAg negative). This is a marked decrease compared to the hepatitis B prevalences reported in previous studies. Among rural migrant mine workers, for example, HBsAg prevalence was 9.9%, and was 10.1% among children 0-6 years of age in the Eastern Cape Province. CONCLUSION: The low-dose, plasma derived hepatitis B vaccine, which is affordable to most developing countries, was very successful in controlling endemic hepatitis B infection, where the virus is predominantly spread by horizontal transmission among infants and young children. PMID- 12075364 TI - Prevalence of nutritional wasting in populations: building explanatory models using secondary data. AB - OBJECTIVE: To understand how social context affects the nutritional status of populations, as reflected by the prevalence of wasting in children under 5 years of age from Africa, Latin America, and Asia; to present a systematic way of building models for wasting prevalence, using a conceptual framework for the determinants of malnutrition; and to examine the feasibility of using readily available data collected over time to build models of wasting prevalence in populations. METHODS: Associations between prevalence of wasting and environmental variables were examined in the three regions. General linear mixed models were fitted using anthropometric survey data for countries within each region. FINDINGS: Low birth weight (LBW), measles incidence, and access to a safe water supply explained 64% of wasting variability in Asia. In Latin America, LBW and survey year explained 38%; in Africa, LBW, survey year, and adult literacy explained 7%. CONCLUSION: LBW emerged as a predictor of wasting prevalence in all three regions. Actions regarding women's rights may have an effect on the nutritional status of children since LBW seems to reflect several aspects of the conditions of women in society. Databases have to be made compatible with each other to facilitate integrated analysis for nutritional research and policy decision-making. In addition, the validity of the variables representing the conceptual framework should be improved. PMID- 12075365 TI - The contribution of breast milk to toddler diets in western Kenya. AB - OBJECTIVE: To understand the relative contributions of breast milk and the weaning diet to overall nutrient intake, with a view to designing and implementing appropriate programmes to improve complementary feeding in developing countries. METHODS: Complementary food intake was measured in a sample of 250 toddlers (mean baseline age: 13.9 +/- 2.4 months) using 24-h dietary recall interviews administered once every 3 weeks over a 6-month period. Breast milk intake over a 24-h period was measured using the test-weighing method in a subsample of 50 children. Regression effects of age and sex on observed milk intakes were estimated and imputed to the whole sample to estimate mean intake over the observation period. Total energy and nutrient intakes were evaluated for adequacy with reference to published estimates of toddler requirements. FINDINGS: Total energy intake (1029 kcal/day) was adequate, with breast milk supplying an average of 328 kcal/day (32%), but vitamin A, riboflavin, calcium, iron and zinc intakes were below current estimates of required intakes. Observed limitations in nutrient intake were consistent with the finding that almost half of the toddlers were stunted. The prevalence of wasting was 6% at baseline and 4% at final assessment. Although food consumption increased when breastfeeding stopped, it could not fully compensate for the fat and vitamin A previously supplied by breast milk. CONCLUSIONS: The nutritional role of mother's milk in the second year is inversely related to the adequacy of the complementary diet. In this study, breast milk was an irreplaceable source of fat and vitamin A. When the weaning diet is inadequate for key nutrients because of low intake or poor bioavailability, breast milk assumes greater nutritional significance in the second year of life but does not guarantee adequate nutrient intakes. PMID- 12075366 TI - Gender and use of cataract surgical services in developing countries. AB - OBJECTIVE: To determine, from the existing literature, cataract surgical coverage rates by sex and the proportion of cataract blindness that could be eliminated if women and men had equal access to cataract surgical services. METHOD: Methodologically sound population-based cataract surveys from developing countries were identified through a literature search. Cataract surgical coverage rates were extracted from the surveys and rates for women were compared to those for men. Peto odds ratios were calculated for each survey and a meta-analysis of the surveys was performed. FINDINGS: From a literature review and meta-analysis of cataract surveys in developing countries, we found that the cataract surgical coverage rate was 1.2-1.7 times higher for males than for females. For females, the odds ratio of having surgery, compared to males, was 0.67 (95% confidence interval (CI): 0.60- 0.74). Despite their lower coverage rate, females accounted for approximately 63% of all cataract cases in the study populations, and if they received surgery at the same rates as males, the prevalence of cataract blindness would be reduced by a median of 12.5% (range 4-21%). CONCLUSION: Closing the gender gap could thus significantly decrease the prevalence of cataract blindness, and gender-sensitive intervention programmes are needed to improve cataract surgical coverage among females. PMID- 12075367 TI - Estimating human rabies mortality in the United Republic of Tanzania from dog bite injuries. AB - OBJECTIVE: To make quantitative predictions about the magnitude of underreporting of human rabies deaths in the United Republic of Tanzania. METHODS: Human rabies deaths were estimated by using a series of probability steps to calculate the likelihood of rabies developing after the bite of a suspected rabid dog, incorporating field data on the incidence of animal bite injuries, the accuracy of rabies recognition, the distribution of bite wounds, and post-exposure treatment. FINDINGS: Predicted human rabies mortality was estimated to be (a) 1499 deaths per year (95% confidence interval 891-2238), equivalent to an annual incidence of 4.9 (2.9-7.2) deaths/100,000, when active surveillance data on bite incidence were used, and (b) 193 deaths per year (32-409), corresponding to an annual incidence of 0.62 (0.1-1.32) deaths/100,000, when national bite statistics were used. The annual mean number of rabies deaths officially recorded for the same period was 10.8 (7.7-14.0). CONCLUSION: In the United Republic of Tanzania, cases of rabies in humans have been greatly underreported. Dog bite injuries are an accessible source of epidemiological data that may be used to estimate the public health burden of rabies and to monitor epidemiological trends in developing countries. PMID- 12075368 TI - Can post-eradication laboratory containment of wild polioviruses be achieved? AB - The purpose of containment is to prevent reintroduction of wild polioviruses from laboratories into polio-free communities. In order to achieve global commitment to laboratory containment the rationale should be clear and compelling; the biosafety levels should be justified by the risks; and the objectives should be realistic. Absolute containment can never be assured. Questions of intentional or unintentional non-compliance can never be wholly eliminated. Effective laboratory containment is, however, a realistic goal. Prevention of virus transmission through contaminated laboratory materials is addressed by WHO standards for biosafety. The principal challenge is to prevent transmission through unrecognized infectious laboratory workers. Such transmission is possible only if the following conditions occur: infectious and potentially infectious materials carrying wild poliovirus are present in the laboratory concerned; a laboratory operation exposes a worker to poliovirus; a worker is susceptible to an infection that results in the shedding of poliovirus; and the community is susceptible to poliovirus infections. At present it is difficult to envisage the elimination of any of these conditions. However, the risks of the first three can be greatly reduced so as to create a formidable barrier against poliovirus transmission to the community. Final biosafety recommendations must await post-eradication immunization policies adopted by the international community. PMID- 12075369 TI - Questioning the indicators of need for obstetric care. AB - The difficulties in measuring maternal mortality have led to a shift in emphasis from indicators of health to indicators of use of health care services. Furthermore, the recognition that some women need specialist obstetric care to prevent maternal death has led to the search for indicators measuring the met need for obstetric care. Although intuitively appealing, the conceptualization and definition of the need for obstetric care is far from straightforward, and there is relatively little experience so far in the use and interpretation of indicators of service use or need for obstetric care. In this paper we review indicators of service use and need for obstetric care, and briefly discuss data collection issues. PMID- 12075371 TI - Blaming malaria rise on climate change is simplistic. PMID- 12075370 TI - What can be done about the private health sector in low-income countries? AB - A very large private health sector exists in low-income countries. It consists of a great variety of providers and is used by a wide cross-section of the population. There are substantial concerns about the quality of care given, especially at the more informal end of the range of providers. This is particularly true for diseases of public health importance such as tuberculosis, malaria, and sexually transmitted infections. How can the activities of the private sector in these countries be influenced so that they help to meet national health objectives? Although the evidence base is not good, there is a fair amount of information on the types of intervention that are most successful in directly influencing the behaviour of providers and on what might be the necessary conditions for success. There is much less evidence, however, of effective approaches to interventions on the demand side and policies that involve strengthening the purchasing and regulatory roles of governments. PMID- 12075372 TI - China sets up Centres for Disease Control and Prevention. PMID- 12075373 TI - Court orders South Africa to treat pregnant HIV-positive women with nevirapine. PMID- 12075374 TI - Could antibiotics cure river blindness? PMID- 12075375 TI - UN to help developing countries assess safety of GM crops. PMID- 12075381 TI - Heparin-induced thrombocytopenia: pathophysiology and management. AB - Immune-mediated heparin-induced thrombocytopenia (HIT) is a well-defined syndrome. Clinical criteria (thrombocytopenia, resistance to heparin anticoagulation, or thromboses during heparin therapy) are defined, and serologic diagnostic tests are available. Earlier recognition of HIT syndrome has allowed for significant advances in therapy, leading to marked reductions in morbidity and mortality from HIT syndrome. This review addresses the epidemiology, pathobiology, and management of HIT syndrome PMID- 12075382 TI - Prophylactic antibiotics prior to bacteremia decrease endovascular graft infection in dogs. AB - Endovascular placement of vascular stent grafts in the aorta and peripheral vessels has become a prominent tool in the armamentaria of the vascular surgeon. Despite, several reports of stent graft infection, no current guidelines exist regarding the administration of antibiotics prior to episodes of potential bacterial seeding. We sought to clarify the role of prophylactic antibiotics in preventing stent graft infection after the parenteral administration of Staphylococcus aureus (S. aureus) at various intervals following device placement. A stent graft device was constructed from a 4 mm thin-walled polytetrafluoroethylene (PTFE) graft attached to the outside of a balloon expandable 394-Palmaz stent (Johnson and Johnson Interventional Systems, Warren, NJ). It was then inserted into the common iliac artery through an 11F peal-away sheath placed in the femoral artery. Sixty grafts were placed into 30 dogs. There were 5 groups of equal number (groups A-E). In group A, six dogs received intravenous injection of 3 cc x 104 CFU (colony forming units), biotype 31375 S. aureus, 1 day after stent graft implantation. An identically treated group B received antibiotic prophylaxis (1 gm cefazolin 30 minutes prior to bacterial challenge). Group C received bacterial injection 7 days after graft implantation with no antibiotic prophylaxis. Group D received bacterial injection 7 days after graft implantation with antibiotic prophylaxis. A control group E received no antibiotics and was not infected. All infected animals were sacrificed 7 days following bacterial challenge and the stent graft complex cultured. One half of the control group was sacrificed at 7 days and the other half at 14 days. The overall stent graft patency was 90%. Four of the six graft occlusions occurred in group A. Eleven of 12 (92%) dogs cultured S. aureus (biotype 31375) from the explanted stent graft complex. Two localized perforations occurred at the site of the infected complex. In group B, C, and D, no explanted graft complex cultured S. aureus. One graft occluded in group C and D. No stent graft in the control (group E) cultured S. aureus. A stent graft infection model can be consistently produced. In the canine model, the stent graft is more susceptible to infection in the early postoperative period and becomes less susceptible to bacterial seeding at one week after implantation. The authors recommend the use of prophylactic antibiotics in the prevention of endovascular graft infections in the early postoperative period during times when bacterial seeding may occur. They postulate that pseudointima formation during graft incorporation into the vessel wall may be responsible for the resistance to infection. PMID- 12075383 TI - The treatment of varicose veins with external stenting to the saphenofemoral junction. AB - Presented are the experiences with 1,516 external valvular stents (Venocufft and Venocuff II) implanted at the saphenofemoral junction (SFJ) between 1985 and 2000. To assess the applicability of the procedure it was found that the appropriate implantation was performed in 34% of 310 consecutive venous procedures. To assess patient preference between external valvular stenting and simultaneous contralateral stripping, 56 consecutive patients were followed up at 3 months postoperatively. Four percent preferred stripping, 4% had no preference, and 92% preferred Venocuff IItrade mark implantation. Competence at the SFJ with specific duplex ultrasound indicators was 94% at 3 months (n = 100) and 90% at 4.8 years (n = 107). Minimal residual reflux (less than 50 mL/minute with maximum Valsalva) was present in the remainder but did not produce symptoms and very rarely progressed over the mean time of 5 years. The internal diameter (ID) of the long saphenous vein (LSV), 3 cm distal to the SFJ, changed from 7.6 +/- 2.3 mm to 4.9 +/- 1.1 mm (p < 0.001) and at the knee from 6.9 +/- 1.9 mm to 3.7 +/- 1.0 mm (p < 0.001). Patients presenting with underlying deep venous disease began with significantly higher ID, ie, 9.0 +/- 2.1 mm at the upper end of the LSV and 7.1 +/- 2.0 mm at the knee, but postoperatively the IDs reverted to those of postoperative patients with a normal deep venous system. To assess patients with recurrences, 366 limbs had simultaneous stripping and contralateral SFJ repair with the Venocuff II. Of these 33 (9%) had recurrences at 4.9 years, 82% of them on the stripping side, and on the repair side half of the recurrences had a competent SFJ (9%). Limbs with an incompetent lateral or anterior accessory system, with an incompetent SFJ (168), were compared with 11 matched randomized controls where stripping was performed. The recurrence rate was 1.2% versus 36% on the strip side. The follow-up for these cases was 6.4 years. Pregnancy (n = 14) produces a high recurrence rate, but stripping and valve repairs were not significantly different, ie, despite small numbers, there was a very strong tendency toward higher recurrence rates on the stripped side. The complication rate was small and the cost of the device is low. The method allows a repairable nonablative approach that can be offered in patients where no other surgical treatment can or should be offered. External stenting to the SFJ is the preferred option for early to moderate varicose veins involving the LSV where the clinical and ultrasonic indicators have been fulfilled. PMID- 12075384 TI - Assessment of gut mucosal perfusion and colonic tissue blood flow during abdominal aortic surgery with gastric tonometry and laser Doppler flowmetry. AB - The objective of this study was to investigate the effect of infrarenal aortic cross-clamping and unclamping on gut mucosal perfusion by gastric tonometry and on sigmoid colonic tissue blood flow by laser Doppler flowmetry during abdominal aortic surgery. This was a prospective before-and-after intervention comparison study in a university hospital of 8 male patients, aged 57-87, undergoing elective infrarenal abdominal aortic surgery. Each patient was pretreated with ranitidine. Following general anesthesia, a nasogastric tonometer was inserted into the stomach. The balloon of the tonometer was filled with 2.5 mL of normal saline for gas tension and pH analysis. This process was repeated before and after aortic cross-clamping and unclamping. Gastric mucosal pHi was calculated with the Henderson-Hasselbalch equation from the arterial Hco3- and the tonometrically measured mucosal Pco2. A laser Doppler flow probe was placed in contact with the serosa of the sigmoid colon against the mesentery after the abdomen was opened. Sigmoid colonic tissue blood flow (SCBF) was assessed by the laser Doppler flowmeter. Gastric mucosal pHi by gastric tonometry and colonic tissue blood flow by laser Doppler flowmetry were measured before and after aortic cross-clamping and unclamping. Gastric mucosal pHi decreased significantly 30 minutes after aortic cross-clamping (7.37 +/-0.07) (p < 0.01), 60 minutes after aortic cross-clamping (7.39 +/-0.08) (p < 0.05), and 30 minutes after aortic unclamping (7.37 +/-0.08) (p < 0.01), compared with pHi before aortic cross-clamping (7.50 +/-0.06). Gastric mucosal pHi increased to the original level 60 minutes after aortic unclamping (7.46 +/-0.08). Since a gastric mucosal pH above 7.35 is considered normal, these mean values of pHi were clinically insignificant. However, gastric mucosal pHi decreased below 7.32 in 5 patients during abdominal aortic surgery. Gastric mucosal pHi decreased further to 7.30 in 1 patient following aortic cross-clamping and below 7.30 in 3 patients 30 minutes after aortic unclamping. SCBF decreased significantly after aortic cross-clamping (28.1 +/-4.8 mL/min/100 g) compared with the value before aortic cross-clamping (51.9 +/-11.3 mL/min/100 g) (p < 0.01). Following aortic unclamping, SCBF returned to 41.7 +/-7.4 mL/min/100 g. It is concluded that transient episodes of significant intestinal mucosal ischemia may have been encountered occasionally in patients undergoing abdominal aortic surgery, but a sigmoid colonic tissue blood flow of 41.7 +/-7.4 mL/min/100 g was sufficient to prevent postoperative ischemic colitis regardless of whether there was ligation or no ligation of inferior mesenteric artery among the studied population since none of the patients developed clinically significant ischemic colitis. PMID- 12075385 TI - Comparison of the use of enoxaparin versus unfractionated heparin in patients undergoing lower extremity revascularization. AB - Present available studies suggest that heparin prevents early vessel thrombosis in the immediate postoperative (POP) period after lower extremity bypass. Long term anticoagulation with warfarin has also been used in these patients, based on its beneficial effect in preventing long-term graft failure. To compare the effectiveness between unfractionated heparin (UH) and low-molecular-weight heparin (enoxaparin) when used as transitional therapeutic POP anticoagulant therapy, a prospective study was performed. Seventy consecutive patients undergoing lower extremity bypass were studied. The initial 35 patients were started on a drip of UH 6 hours POP. The next 35 consecutive patients were treated with enoxaparin, the first dose starting 6 hours POP. All patients underwent subsequent anticoagulation with oral warfarin. There were no statistically significant differences between the two groups with regard to demographics, percentage of above/below the knee bypasses, type of conduit used, emergency operations, and overall complications. There was a significant difference in the length of stay (p = 0.05) in favor of the enoxaparin group, which also reflected a decreased overall cost in this group. In conclusion, POP anticoagulation with enoxaparin is as safe end effective as UH in patients undergoing lower extremity revascularization. Enoxaparin was associated with a decreased overall length of stay and hospital costs. PMID- 12075386 TI - Does the efficacy of dorsalis pedis artery bypasses vary among diabetic patients of different ethnic backgrounds? AB - Hispanic patients suffer from a high rate of leg amputations, far beyond what would be expected from the high prevalence of diabetes in this population. This raises questions about the efficacy of bypass operations across ethnic lines. We focused this review on dorsalis pedis bypasses, as these are frequently performed in diabetic patients. We compared outcomes between Hispanics and non-Hispanics and sought to identify factors predictive of failure or complications. The authors conducted a retrospective review of 144 dorsalis pedis bypasses in 106 men and 29 women with a mean age of 62 years. Eighty-two percent were Hispanic; 96% of cases were done for tissue loss, and 4% for rest pain. Twenty-five percent of patients experienced perioperative complications; these were more frequent in non-Hispanics than Hispanics (40% vs 22%, p = 0.05). The most frequent complications were wound related (11%). The 30-day mortality was 1.5% and 30-day graft thrombosis was 5%. Follow-up ranged from 1 to 62 months and averaged 12 months. Eighty-one percent of the limbs at risk were saved, although 36% of cases required minor foot amputations. Estimated primary graft patency was 68% at 30 months. The 30-month Kaplan-Meier curves for primary patency, assisted patency, and limb salvage were not statistically different between Hispanics and non Hispanics (p > 0.4). Grafts that remained patent had higher duplex-derived intraoperative flow velocities in the dorsalis pedis artery than grafts that eventually failed (121 +/-69 vs 74 +/-26 cm/sec, p = 0.02). In grafts that remained patent, dorsalis pedis velocity decreased from the perioperative period to the 8 to 12 weeks time point, whereas no change was seen in grafts that eventually failed (mean decline of 48 +/-76 vs 1 +/-58 cm/sec, p = 0.02). No other factors were predictive of graft failure. The results of dorsalis pedis bypass in Hispanic patients compare favorably to those seen in other ethnic groups. This suggests that other factors must account for the high amputation rates seen in Hispanics, such as a frequent occurrence of nonreconstructible disease or unaccounted for cardiovascular risk factors. The usefulness of duplex derived flow velocities in the dorsalis pedis to predict long-term graft patency warrants further investigation. PMID- 12075387 TI - The Ehlers-Danlos specter revisited. AB - Ehlers-Danlos type IV is a major concern to vascular surgeons because it is often associated with spontaneous hemorrhage from arteries containing decreased type III collagen. Five members of a family with Ehlers-Danlos type IV and a review of another family of five with Ehlers-Danlos type IV are reported. Evaluation of the recent family included clinical evaluation as well as assay of collagen production. The age range of the three involved females and two males was 7 to 52 years. The father of the affected family had a spontaneous colon perforation at age 39. His son, at age 27, had a spontaneous rupture of the iliac artery. Revascularization was accomplished with difficulty. His daughter had a large cerebral bleed. Two granddaughters, ages 7, have not had any bleeding or aneurysmal events. The amount of type III collagen was only 10% of normal in the patient with the iliac artery rupture. The three females all exhibited similarly low levels of type III collagen. The father's type III collagen level was not sufficiently low to confirm Ehlers-Danlos type IV, although he had a spontaneous colon perforation. In the other Ehlers-Danlos type IV family of five, the three surviving members had type III collagen levels as low as 5% of normal. Two family members died after spontaneous iliac rupture at ages 24 and 33. Both families exhibited an autosomal dominant inheritance pattern. Ehlers-Danlos type IV remains a challenging problem for vascular surgeons. It is transmitted as an autosomal dominant inheritance with a high degree of penetrance. Spontaneous arterial and intestinal perforations should alert the clinician to the possibility of Ehlers-Danlos type IV. Patients should be evaluated noninvasively. Arterial repairs may not be successful in these patients because the vessels are extremely friable. Assays of collagen production are advisable in establishing the diagnosis. PMID- 12075388 TI - Ischemic jejunal stricture following endovascular abdominal aortic aneurysm repair -- a case report. AB - Endograft repair has rapidly become an alternative to conventional open repair of abdominal aortic aneurysms. Various trials continue to show decreased morbidity when compared to open repair. However, as with any new procedure, complications specifically related to this technique are being described. Herein, we report a case of an isolated ischemic jejunal stricture presenting as a small-bowel obstruction secondary to cholesterol emboli following endograft repair of an abdominal aortic aneurysms. PMID- 12075389 TI - A comparison of patch angioplasty and stenting for axillary venous stenoses of thrombosed hemodialysis grafts. AB - The efficacy of balloon angioplasty with stent placement is compared to surgical patch angioplasty for thrombosed upper arm hemodialysis grafts with stenotic lesions at the venous anastomosis. Patients with thrombosed hemodialysis grafts terminating at the axillary vein were reviewed. Thirty-eight stents were placed after thrombectomy and venous balloon angioplasty in 26 patients. Fifteen case matched controls underwent 23 polytetrafluoroethylene patch angioplasties for similar lesions. Kaplan-Meier survival analysis was used for statistical comparison. Primary patency for stent placement compared with patch angioplasty was not statistically different at any time through 1-year follow-up (37% vs 55% at 3 months, 25% vs 45% at 6 months, and 25% vs 15% at 12 months, respectively; p = 0.37). Secondary patency rates were slightly better for patch angioplasty compared with stent placement (78% vs 59% at 3 months, 78% vs 48% at 6 months, and 54% vs 32% at 12 months, respectively) however these differences were not statistically significant (p = 0.13). There was no difference in complication rates between groups. Sustained patency is poor for both angioplasty with stent placement and surgical patch angioplasty when revising thrombosed dialysis grafts with anastomotic axillary vein stenosis. These data suggest slightly better patency for the routine use of patch angioplasty for these lesions. However, the endovascular approach appears to be a reasonable alternative when surgical exposure is difficult. PMID- 12075390 TI - Safety and cost savings of endovascular procedures: are outpatient interventions feasible when combined with open surgery? AB - As increasing experience and comfort with endovascular interventions performed in an outpatient setting has occurred, the safety and cost effectiveness of performing these procedures without an overnight stay were analyzed, especially when endovascular procedures were combined with open vascular operations requiring an arteriotomy and surgical closure. Ninety patients underwent endovascular procedures alone or concomitantly with open, minor vascular operations to salvage a failing graft between February 1994 and June 1999. Patients undergoing endovascular interventions during primary lower extremity bypass or other major surgical procedures were not included in this review because they were not candidates for outpatient procedures. Balloon angioplasty alone (79) or angioplasty with stent placement (11) was performed to treat stenoses in 50 failing grafts, 16 iliac, 14 femoral, 5 tibial, and 5 axilla/subclavian arteries. A significant increase in outpatient procedures was accomplished as more experience was garnered with these techniques: 19% (8/42) between 1994 and 1996 vs 57% (28/48) between 1997 and 1999 (p = 0.001). Age and comorbidity did not play a role in determining the need for admission because there were no significant differences in patients with diabetes mellitus, hypertension, smoking, or hyperlipidemia and those admitted or discharged the same day (p > 0.05). Patients admitted for overnight observation tended to have longer mean operative times and more complex revascularizations than outpatients (110 vs 69 min, respectively; p < 0.0001). Twenty-seven patients underwent surgical exposure of the access vessel: 63% (17) were admitted and 37% (10) were discharged the same day. Sixty-three patients underwent a percutaneous procedure: 42% (27) were admitted and 58% (37) were discharged the same day. Outpatients were more likely to receive only local anesthesia (83%; 30/35) compared to patients admitted overnight (67%; 36/53); the remaining patients received spinal or epidural anesthesia. Complications included graft thrombosis within 30 days in 6% (5/90) of patients and arterial graft infection in 2% (2). No patient required surgery for bleeding. The average charges for outpatient interventions were $1980 compared to $10,026 for patients who stayed overnight (p < 0.0001). As vascular surgeons become more experienced and comfortable with outpatient endovascular procedures, especially when performed in combination with open minor vascular surgery, significant cost savings can be realized without sacrificing patient safety. Even when open surgical exposure is planned, patients should be instructed preoperatively to anticipate discharge the day of their procedure to minimize resistance to this strategy. PMID- 12075391 TI - Iliac graft access to facilitate endovascular repair of a ruptured thoracic aortic aneurysm -- a case report. AB - Arterial access for endovascular aortic aneurysm repair is usually gained through a common femoral artery approach. In small femoral arteries this can be difficult or even impossible owing to the large size of the introduction sheath of the delivery system. In such cases the iliac arteries or the abdominal aorta can be used for vascular access, although, in heavily calcified arteries,even this can be hazardous. The authors report an 81-year-old woman with a contained rupture of the thoracic aorta in whom a polyester graft was used to facilitate vascular access to the common iliac artery for a safe two-stage endovascular repair of the aneurysm. PMID- 12075392 TI - Aneurysm of the dorsalis pedis artery -- a case report. AB - Aneurysms of the pedal arteries are uncommon; however, they can be identified upon clinical exam and confirmed by angiogram and color-flow duplex scan. Surgical treatment options include ligation or primary repair. The authors present a case of an aneurysm of the dorsalis pedis artery in a diabetic patient. Primary repair of the aneurysm was accomplished using a venous autograft patch. The postoperative course was uneventful and the artery remains patent in follow up. PMID- 12075393 TI - Antiarrhythmic drugs in cardiac arrest resuscitation: intravenous amiodarone or intravenous lidocaine? PMID- 12075394 TI - Spontaneous adverse event reports of serious ventricular arrhythmias, QT prolongation, syncope, and sudden death in patients treated with cisapride. AB - CONTEXT: Adverse cardiac events, typically long QT syndrome, have been reported in patients treated with the gastrointestinal prokinetic agent cisapride. OBJECTIVE: To analyze cases of cisapride-associated long QT syndrome and ascertain the degree of confidence in the diagnosis of long QT syndrome and the presence of risk factors. DESIGN: Review of all cases reported to the manufacturer, health authorities, or in the medical literature as of October 1999. METHODS: Before review, specific definitions and a classification scheme were agreed upon. Each case was classified by confidence in the long QT syndrome diagnosis and the presence of recognized risk factors (cofactors) or other medical conditions that were listed as contraindications in the June 1998 United States labeling (labeled conditions). RESULTS: Of 574 cases reviewed, 391 cases of long QT syndrome or isolated QT prolongation were confirmed. Most of these were classified as long QT syndrome with high (145, 37%) or medium (92, 23.5%) confidence in the diagnosis. Recognized cofactors were present in 262 (67%) cases. The proportion of cases with recognized cofactors rose with confidence in the diagnosis (P < 0.001) from 42.2% in the low-confidence group to 68.5% and 82.1% in the medium- and high-confidence groups, respectively. Conversely, the proportion of cases with other labeled conditions decreased with confidence in the diagnosis, from 33.3% for low confidence to 13% and 8.3% for medium and high confidence, respectively. Analysis of cases and prescribing data showed reporting rates decreased in studied months in 1999 compared with the average study period. CONCLUSION: In most cases with high or medium confidence in the diagnosis of cisapride-associated long QT syndrome, recognized cofactors for long QT syndrome were present. The risk of cisapride-related long QT syndrome may be minimized by avoiding cofactors. PMID- 12075395 TI - Use of statins and aspirin to reduce risks of cardiovascular disease. PMID- 12075396 TI - Effects of diltiazem and esmolol on cycle length and spontaneous conversion of atrial fibrillation. AB - BACKGROUND: Calcium channel blocking agents have been shown to prolong the duration of atrial fibrillation. This study compared the effects of intravenous diltiazem and esmolol on the cycle length and conversion rate of pacing-induced atrial fibrillation. METHODS AND RESULTS: In 41 adults without structural heart disease, atrial fibrillation was induced by rapid atrial pacing. After 3 minutes, either diltiazem (n = 13), esmolol (n = 15), or saline (n = 13) was infused. In the diltiazem group, the atrial fibrillation cycle length shortened by a mean of 43 milliseconds and became significantly shorter than in the control group, while the atrial fibrillation cycle length in the esmolol group did not change. Spontaneous termination of atrial fibrillation occurred significantly less often in the diltiazem group (23%) than in the esmolol (67%, P < 0.05) or placebo groups (77%, P = 0.01). CONCLUSIONS: Intravenous diltiazem shortens the atrial fibrillation cycle length and lowers the probability of spontaneous conversion of recent-onset atrial fibrillation to sinus rhythm. These results suggest that the use of diltiazem for acute rate control may unwittingly prolong the duration of recent-onset atrial fibrillation. PMID- 12075397 TI - Downregulation of immunodetectable atrial connexin40 in a canine model of chronic left ventricular myocardial infarction: implications to atrial fibrillation. AB - BACKGROUND: The substrate(s) for atrial fibrillation associated with chronic left ventricular myocardial infarction remain poorly defined. Since atrial connexin40 has a rapid turnover rate and may cause atrial fibrillation, we hypothesized that chronic left ventricular myocardial infarction downregulates atrial Connexin40 and increases atrial fibrillation vulnerability. METHODS AND RESULTS: The left anterior descending coronary artery was occluded distal to the first diagonal branch in five dogs and studied 7 weeks later. Five dogs with no left anterior descending coronary artery occlusion served as control. Vulnerability to atrial fibrillation was tested by burst atrial stimulation (50 milliseconds for 3 seconds). Atrial fibrillation was induced in all myocardial infarction dogs, lasting from 20 seconds to several minutes. In contrast, only rapid repetitive activity and short-lasting atrial fibrillation (< 5 seconds) could be induced in control dogs. The mean refractory periods of epicardial RA and LA appendages were not significantly different in the two groups. Mean left ventricular myocardial infarction size was 17 +/- 4% of the left ventricle. Histologic analyses showed no signs of atrial ischemic injury or interstitial fibrosis in either group. Atrial myocyte diameter measured at the level of the nuclei of longitudinally sectioned myocytes was not significantly different in the two groups (10.1 +/- 1.2 microm vs. 10.2 +/- 1.2 microm; P = 0.3). Atrial Connexin40 (both left and right atria) in the left ventricular myocardial infarction group was highly heterogeneous and had significantly smaller total area stained than in the control (0.48 +/- 0.09% vs. 0.82 +/- 0.13%; P < 0.01). CONCLUSIONS: Chronic left ventricular myocardial infarction downregulates immunodetectable atrial Connexin40, a property that might contribute to the increased atrial fibrillation vulnerability in this model. PMID- 12075398 TI - Beneficial effects of therapy on the progression of structural remodeling during healing after reperfused and nonreperfused myocardial infarction: different effects on different parameters. AB - BACKGROUND: Structural left ventricular remodeling after myocardial infarction is a complex process with several pathophysiologic descriptors that can be modified by pharmacotherapy. However, the possibility that different classes of antiremodeling agents might exert different effects on different remodeling parameters after reperfused and nonreperfused myocardial infarction has not been systematically studied. METHODS AND RESULTS: We measured detailed left ventricular remodeling parameters in vivo (echocardiograms) repeatedly over 6 weeks and ex vivo (planimetry) at 6 weeks after myocardial infarction in 36 dogs randomized (factorial design) after reperfused or nonreperfused myocardial infarction to 6 weeks of twice daily oral therapy with the calcium channel blocker amlodipine (5 mg), the angiotensin-converting enzyme inhibitor enalapril (5 mg) or placebo, and 18 matching sham or control animals. Compared to placebo and control groups over 6 weeks, both agents reduced left ventricular loading and limited overall remodeling in both reperfused and nonreperfused groups, but there were pertinent differences. Enalapril limited the increase in left ventricular asynergy in the reperfused group. Both enalapril and amlodipine limited infarct zone thinning in the nonreperfused groups but increased infarct zone thinning in the reperfused groups, despite preserved infarct zone collagen with amlodipine. Enalapril decreased left ventricular diastolic volume and mass more than amlodipine in the reperfused group and increased left ventricular ejection fraction in the nonreperfused group. Both agents limited regional and global shape deformation in reperfused and non-reperfused groups. Diastolic wall stress in the infarct zone decreased with amlodipine, and increased with enalapril and reperfusion. CONCLUSIONS: Different antiremodeling therapies may exert different effects on different remodeling parameters during healing after reperfused myocardial infarction. Significant interactions occur during reperfusion. More than one variable may be needed for the comprehensive assessment of the antiremodeling efficacy of different therapies. PMID- 12075399 TI - Mechanism of the positive inotropic effect of lysophosphatidic acid in rat heart. AB - BACKGROUND: Lysophosphatidic acid, a bioactive phospholipid, is mainly released from the activated platelets. The concentration of lysophosphatidic acid in serum is elevated under conditions such as ischemia, hypertension, and thrombosis; however, its effect on cardiac function, as well as the mechanisms of its action, have not been fully understood. METHODS AND RESULTS: Cardiovascular effects of lysophosphatidic acid were studied in vivo in rats as well as in vitro by using the isolated perfused heart and cardiomyocyte preparations. Intravenous injection of lysophosphatidic acid (2.8 to 14 microg/100 g body wt) increased the left ventricular systolic and diastolic pressures, rate of pressure development, and rate of pressure decay in rats. The positive inotropic effect of lysophosphatidic acid in vivo was not affected by the blockers of angiotensin II, endothelin-1, or adrenergic receptors, but this action was abolished by pretreatment with neurokinin type 1 receptor antagonist (L703606) as well as Ca2+-channel antagonist (verapamil). In the isolated heart, lysophosphatidic acid (1-10 microM) had no significant effect on cardiac function but higher concentrations (20-50 microM) elevated the left ventricular end diastolic pressure, significantly. Lysophosphatidic acid (1-30 microM) neither showed any effect on the basal intracellular concentration of free Ca2+ nor modified the KCl-induced increase in [Ca2+]i in freshly isolated cardiomyocytes. CONCLUSIONS: These results indicate that lysophosphatidic acid stimulated heart function under in vivo but not in vitro conditions. The positive inotropic effect of lysophosphatidic acid in vivo may be indirectly mediated by the activation of neurokinin type 1 receptors. PMID- 12075400 TI - Diabetes, heart rate, and mortality. AB - A large body of evidence indicates that a persistently high heart rate is associated with a significant risk for higher mortality and sudden death in individuals with a variety cardiovascular disorders, as well as in the general population. Heart rates elevated beyond a certain threshold have been found to be a risk factor for mortality in patients with hypertension, in survivors of myocardial infarction, and in patients with impaired cardiac function. Conversely, a naturally slow heart rate, or one that is slow by virtue of sympathetic blockade induced by pharmacologic agents, may result in longer survival. This is particularly evident in the case of beta-adrenergic blocking drugs, especially in patients after myocardial infarction and in those with acute as well as chronic cardiac failure, a syndrome in which there is a complex neurohormonal disturbance with elevated heart rate. Persistently elevated heart rate is also a feature of diabetes mellitus associated with autonomic neuropathy. Whether this also constitutes an independent risk factor for sudden and augmented mortality is not well defined. In this review, the data on the role of increased heart rate as a risk factor for mortality are examined in the context of other factors that may have therapeutic implications. PMID- 12075401 TI - Diagnosis of gastrointestinal stromal tumors: a consensus approach. AB - As a result of major recent advances in understanding the biology of gastrointestinal stromal tumors (GIST), specifically recognition of the central role of activating KIT mutations and associated KIT protein expression in these lesions, and the development of novel and effective therapy for GISTs using the receptor tyrosine kinase inhibitor STI-571, these tumors have become the focus of considerable attention among pathologists, clinicians, and patients. Stromal/mesenchymal tumors of the gastrointestinal tract have long been a source of confusion and controversy with regard to classification, line(s) of differentiation, and prognostication. Characterization of the KIT pathway and its phenotypic implications has helped to resolve some but not all of these issues. Given the now critical role of accurate and reproducible pathologic diagnosis in ensuring appropriate treatment for patients with GIST, the National Institutes of Health (NIH) convened a GIST workshop in April 2001 with the goal of developing a consensus approach to diagnosis and morphologic prognostication. Key elements of the consensus, as described herein, are the defining role of KIT immunopositivity in diagnosis and a proposed scheme for estimating metastatic risk in these lesions, based on tumor size and mitotic count, recognizing that it is probably unwise to use the definitive term benign for any GIST, at least at the present time. PMID- 12075402 TI - The expression of EGFR family ligands in breast carcinomas. AB - Expression of EGF, HB-EGF, TGF-alpha, HRG-alpha, HRG-beta1, and HRG-beta3 in 100 frozen breast carcinoma materials was immunohistochemically studied. Among these tumors, 67% were positive for EGF, 53% for HB-EGF, 57% for TGF-alpha, 60% for HRG alpha, 53% for HRG-beta1, and 63% for HRG-beta3 in the neoplastic epithelial cells. No significant associations between expression of the growth factors and clinicopathological features like tumor size, histologic grade, node status, ploidy, ER status, and c-erbB-4 expression were observed, with the exceptions that significant relations were present between EGF expression and tumor size (p = 0.01) and between HRG-beta3 expression and node status (p = 0.02). The expressions of these growth factors showed no association with cancer-specific survival by the Kaplan Meier analysis. PMID- 12075403 TI - Gastrointestinal stromal tumors: an ultrastructural study. AB - Gastrointestinal stromal tumors (GISTs) represent an enigmatic group of lesions of uncertain phenotype and biologic potential. Although earlier studies suggested smooth muscle cells, schwann cells, or neuronal differentiation, more recent evidence indicates that these tumors show phenotypic features that are similar to the interstitial cells of Cajal. Recently, investigators have begun to evaluate these lesions in a site-specific manner and have found that, in addition to morphologic differences between them, their biologic behavior also appears to be linked to their anatomic location. Many of these studies have emphasized the histologic and immunophenotypic features of GISTs in relation to their sites of origin, however, their site-specific ultrastructural characteristics have received little attention in the literature. In this study, we evaluated 34 GISTs (15 gastric, 12 small intestinal, 4 colonic, and 3 omental) for a variety of ultrastructural features in an effort to identify site-specific similarities and differences. Tumors predominantly composed of epithelioid cells were more commonly seen in gastric (60%) and omental (67%) tumors than in those of the small intestine (33%) and colon (0%). Cytoplasmic filaments and intercellular junctions were commonly seen in tumors from all locations, the filaments frequently forming paranuclear aggregates in the epithelioid cells. Tumors from all sites were composed of cells with surface filopodia and interdigitating cell processes, but in tumors of the stomach and omentum the filopodia were usually short and minimally intertwined, whereas those of small and large intestinal GISTs were characteristically long and complex. Basal lamina, though poorly formed, was present only in tumors of gastric and omental origin (13% and 67%, respectively). Pinocytotic vesicles were also seen in tumors from these sites (33% of gastric tumors and 67% of omental lesions) as well as those of the small intestine (17%) and the colon (25%). None of the gastric or omental tumors had microtubules; they were, however, seen in small intestinal (33%) and colonic (25%) stromal tumors. Skenoid fibers were seen in 33% of small intestinal GISTs and 1 metastatic gastric GIST. Overall, gastric and omental tumors have better developed features of myogenic differentiation and have blunt filopodia and minimally intertwined cell processes. Indeed, these 2 groups are indistinguishable ultrastructurally, raising the possibility that the genesis of omental GISTs is similar to that of gastric stromal tumors. Small intestinal stromal tumors have characteristic interdigitating cell processes and numerous elongate filopodia-like structures harboring intercellular junctions as well as microtubules and extracellular skenoid fibers. The constituent cells in colonic stromal tumors, while more reminiscent of small intestinal stromal, were frequently more primitive in appearance. In conclusion, GISTs from different anatomic locations share many overlapping ultrastructural characteristics; however, a few features are distinctive. It is hoped that these findings will aid in their recognition and contribute to the classification of this heterogeneous group of neoplasms. PMID- 12075404 TI - Liver transplantation for alcoholic liver disease with or without hepatitis C. AB - Alcoholic cirrhosis with or without hepatitis C is a common indication for liver transplantation (LT). Comparative post-LT data for the 2 groups are not available. Our aim is to compare the clinicopathologic features of patients with alcoholic liver disease (ETOH) and ETOH/HCV at the time of and after LT and to determine the impact of concomitant hepatitis C virus (HCV) on ETOH patients undergoing LT. A comparative clinical and pathologic analysis at LT and after LT was performed for 56 patients with ETOH and 32 patients with ETOH/HCV. All 88 had cirrhosis at LT. Other native liver features for ETOH and ETOH/HCV, respectively, were: >2+ inflammation 50/56 and 26/32, Mallory's hyalin 12/56 and 6/32, steatosis 9/56 and 7/32, large cell dysplasia 12/56 and 6/32, hepatoma 4/56 and 4/32, iron deposition 24/56 and 12/32; none was statistically significant. The post-LT findings for ETOH and ETOH/HCV were as follows: 1-year survival 93% and 97%; alive 36/56 (419-4,348 days) and 27/32 (488-5,516 days); deaths 20/56 and 5/32; ETOH recurrence 5/56 (all alive) and 3/32 (1 dead); post-LT HCV 4/56 (acquired) and 22/32 (recurrent). Native liver histology in ETOH and ETOH/HCV patients was similar. Post-LT HCV recurrence was common (69%) in ETOH/HCV patients but resulted in death in only 6%. Post-LT ETOH recurrence was uncommon (9%) and progression to liver failure was rare (1.1%). Post-LT outcome for ETOH was excellent, and concomitant HCV did not affect survival. PMID- 12075405 TI - Poorly differentiated carcinomas of the thyroid gland: a review of the clinicopathologic features of a series of 28 cases of a heterogeneous, clinically aggressive group of thyroid tumors. AB - We evaluated the clinicopathologic, immunohistochemical, lectin histochemical, ultrastructural and morphometric characteristics of a series of 28 poorly differentiated carcinomas of the thyroid (PDCT). The 28 tumors were classified according to their predominant growth pattern as insular (n=12), trabecular (n=10) and solid subtypes (n=6). The overall mortality rate was 46%. No significant differences were found among the 3 subtypes. Data obtained by lectin histochemistry, electron microscopy, and morphometry point to a closer relationship of PDCT to follicular than to papillary carcinoma. Our results do not support the subdivision of PDCT into subtypes. PMID- 12075406 TI - Thyroid microcarcinoma: fine-needle aspiration diagnosis and histologic follow up. AB - The increased sensitivity of many imaging modalities (ultrasound, computed tomography scan, magnetic resonance imaging) has resulted in the identification of thyroid nodules, measuring 1 cm or less. Usually these small lesions are regarded as incidental and are not sampled by fine-needle aspiration (FNA). However, some of these lesions undergo FNA because of suspicious radiology findings (multifocality, calcification, etc) or in patients with a history of radiation to the head and neck region. We present FNA findings and histologic follow-up of 39 thyroid nodules that measured 1.0 cm or less. All FNAs were performed under ultrasound guidance. The lesions ranged in size from 0.2 to 1.0 cm. Twenty-two lesions were diagnosed as papillary carcinoma (PTC), 4 as medullary carcinoma (MC), and 13 as suspicious for PTC on FNA. Histologic follow up showed PTC in 35 and MC in 4 cases; 11 PTC were multifocal (31%) and lymph node metastases were present in 8 (16%) cases. Ultrasound-guided FNA is effective in the sampling of thyroid cancers that are 1.0 cm or less. The present study shows that some of these lesions can be clinically significant. PMID- 12075407 TI - The association between chronic lymphocytic thyroiditis and thyroid tumors. AB - An association between lymphocytic thyroiditis and thyroid papillary carcinoma is still controversial. To assess the relationship, a histopathologic analysis of surgically resected thyroid tumors together with the frequency and severity of chronic lymphocytic infiltration of the thyroid among patients with follicular adenoma, follicular carcinoma, and papillary carcinoma was performed. The prevalence of lymphocytic infiltrate, which is indicative of autoimmune thyroiditis, was significantly higher in patients with papillary carcinoma (58%) than in patients with follicular carcinoma (20%) or follicular adenoma (14%). The lymphocytic infiltration within the tumor compared with the severity of thyroiditis in the nontumorous tissue. Therefore, the association between chronic lymphocytic thyroiditis and papillary carcinoma was confirmed. The possibility that an immunologic mechanism involved in the pathogenesis of papillary carcinoma stimulates lymphocytic infiltration in the thyroid tissue through an autoimmune mechanism is suggested. PMID- 12075408 TI - Images in pathology. Infarcted thymoma. PMID- 12075409 TI - Images in pathology. Mammary crystalloids. PMID- 12075410 TI - Papilliferous keratoameloblastoma of mandible: a papillary ameloblastic carcinoma: report of a case with a 6-year follow-up and review of the literature. AB - We report a fourth case of papilliferous keratoameloblastoma, with a 6-year follow-up. A 62-year-old man underwent resection of a right mandibular neoplasm infiltrating bone and soft tissues. Microscopically, there were cysts lined by papillary projections and containing necrotic debris. Cribriform, solid, and tubular patterns were also present. No regional or distant metastases were found. A local recurrence, developed 3 years later, showed a predominance of the lesser differentiated patterns and focal granular cell ameloblastomatous features. The patient died soon after surgery of unrelated causes, with a further local relapse. The tumor coexpressed cytokeratins and vimentin, and S100 protein (the latter focally in the relapse). It was instead unreactive for Alcian blue, actin, maspin, and GFAP. We suggest that papilliferous keratoameloblastoma be renamed "papillary ameloblastic carcinoma." PMID- 12075411 TI - Mediastinal immature teratoma with yolk sac tumor and myelomonocytic leukemia associated with Klinefelter's syndrome. AB - A 14-year-old male presenting with a short history of right subclavicular chest pain was found to have a mediastinal tumor. Hematologic investigations and bone marrow examination showed features of myelomonocytic leukemia. The mediastinal tumor was excised, but the surgery was complicated by massive hemorrhage. The patient's condition deteriorated postoperatively and he died a week later. The histology of the mediastinal tumor showed the typical features of an immature teratoma with a yolk sac tumor. A prominent infiltrate of leukemic blast cells was present within blood vessels and in close proximity to the yolk sac component. The karyotypic analysis of leukemic cells isolated and cultured from the bone marrow showed 50XXY, +8, +21, +iso G-group marker chromosome karyotype. PMID- 12075412 TI - Uterine adenomyoma with uterus-like features: a report of two cases. AB - Two uterine adenomyomas with unusual features are presented. Case 1 was a 43-year old woman who had a 1-year history of menorrhagia and dysmenorrhea. The uterus showed a 3-cm-sized mass attached to the posterior wall of the lower uterine segment. On section, there was a well-circumscribed, submucosal cystic mass with a thick muscular wall. Histologically, the cavities were lined by well-developed secretory-phase endometrium and surrounded by smooth muscle, like a small uterus. Case 2 was a 52-year-old woman who visited the hospital because of itching sensation on the vulva. On physical examination, the mass was found on the left side of the pelvis. The enlarged uterus had an 8-cm-sized protruding mass from the left side of the lower uterine segment. On section, the mass was cystic and solid. Histologically, the cysts were lined by thin basalis-type endometrial glands and stroma, and the solid portion was composed of smooth muscle and fat cells. PMID- 12075413 TI - RET/PTC rearrangements in Hashimoto's thyroiditis. PMID- 12075414 TI - Theoretical Elucidation on Activity Differences of Ten Flavonoid Antioxidants. AB - Theoretical methods including structure-activity relationships (SAR) and quantum chemical calculations were used to elucidate the free radical scavenging activity differences of 10 flavonoid antioxidants. SAR could give a qualitative explanation on the antioxidant activity differences. And a further elucidation was performed by a theoretical parameter, the difference of heat of formation between antioxidant and its free radical (deltaHOF), calculated by Austin Model 1 (AM1) method. Besides, deltaHOF showed a linear correlationship with the logarithm of the relative antioxidant efficiency (lgRAE, r = 0.7523), indicating that the theoretical methods were effective to elucidate the differences of antioxidant activity. On the other hand, the ineffectiveness of another theoretical parameter, the highest occupied molecular orbit (HOMO) energy level, was verified further. PMID- 12075415 TI - Cloning and Expression of Tetanus Toxin Fragment C in E.coli. AB - The fragment C of tetanus toxin was amplified from Clostridium tetani DNA by PCR. This fragment was cloned into expression vector pET-28a(+),under the control of the T7 promoter. Expression of this plasmid in E.coli resulted in the production of a protein consisting of 6xHis of the vector fused to the N-terminal 451 amino acids of tetanus toxin. After induction with 1 mmol/L IPTG, TTC was expressed in E.coli BL21(DE3). The protein product accounted for 8.2% of the bacteria total protein in soluble form, SDS-PAGE and Western blot analysis of TTC recombinant protein confirmed this result. The expression products were also purified by Ni(2+)-IDA-Sephrose 6B column. Immunization of mice with rTTC resulted in the production of antibodies that were able to protect mice against a challenge with tetanus toxin furthermore, rTTC in vivo appeared to be able to undergo retrograde axonal transport. PMID- 12075416 TI - Analysis and Molecular Cloning of Differentially Expressing Genes in Nasopharyngeal Carcinoma. AB - In order to further examine expression of cDNA fragments isolated by cDNA representational difference analysis(cDNA RDA) in nasopharyngeal carcinoma(NPC) biopsies and to clone those deregulated genes associated with NPC, RT-PCR and Northern blot were used to identify the differentially expressed cDNA fragments in NPC biopsies and confirm the transcript length of those genes, then a full length cDNA sequences was cloned and its product was analyzed by bioinformatics. The results showed that AF091521, AF091520, AF152605 and AF091517 cDNA sequences had distinct expression difference between primary cultural normal nasopharyngeal epithelial cell and NPC biopsies, and AF091521, AF091517 genes all had two transcripts whose sizes were 1.5, 2.3 and 1.1, 1.4 kb respectively, while AF091520 and AF152605 gene expressed one transcript only, respectively, whose sizes were 1.6 and 2.2 kb. An AF091517 EST gene, named as NAG11, (GenBank accession number AF170307) was isolated by sequencing one EST clone, which encoded a transmembrane protein of 88 amino acid including three protein ATP binding regions, two protein kinase C phosphorylation sites and two N myristoylation sites. So it is further demonstrated that NPC is a disease with multiple gene alterations NAG11 gene is a candidate of putative tumor suppressor genes associated with NPC, whose down-expression may be involved in the development of NPC and NAG11 gene product may play a role in the transmembrane transport of ATP. PMID- 12075417 TI - Cloning, Expression and Tumor Suppression of Human Endostatin. AB - Human endostatin cDNA was cloned from total RNA of normal Chinese liver cell line L02 by RT-PCR. Endostatin DNA sequence encoded 184 amino acid residues. Five base pairs and 3 amino acid residues are different from that reported, it may be due to interspecies difference. The endostatin cDNA was inserted into the pET-28a(+) containing T7 promoter. The recombinant plasmid was transformed the E.coli BL21(DE3). Recombinant human endostatin was highly expressed as inclusion body when the expression strain BL-ENDO was induced with 1 mmol/L IPTG. Result of SDS PAGE analysis revealed that recombinant human endostatin was accounted for up to 25% of soluble protein in E.coli. Purified and refolded recombinant human endostatin was active in inhibiting tumor growth and metastasis. PMID- 12075418 TI - Purification, Crystal Growth and Preliminary X-ray Analysis of a Phospholipase A(2) from Venom of Agkistrondon acutus. AB - An acidic phospholipase A(2) from Agkistrondon acutus venom has been purified to homogeneity via four steps using CM-Sepharose, two times DEAE-Sepharose, and Mono Q FPLC. The molecular weight of the protein was about 16.5 kD and the isoelectric point was 4.3. The purified enzyme showed a potent inhibitory effect on platelet aggregation induced by ADP in human platelet-enriched plasma. The enzyme was then crystallized by hanging drop diffusion method using 2-methyl-2, 4-pentanediol as a precipitant. Two kinds of single crystals suitable for X-ray crystallographic studies were obtained. X-ray crystallographic analysis showed that both crystal forms belong to monoclinic system and space group P2(1). The cell dimensions of form I crystals were a = 43.48 Aring;, b = 71.49 Aring;, c = 43.85 Aring; and beta = 116.32 deg; Those of form II crystals were a = 49.25 Aring;, b = 38.33 Aring;, c = 70.25 Aring; and beta = 99.20 deg;. Complete diffraction data sets have been collected to medium resolution for the two crystal forms. PMID- 12075419 TI - Inhibitory Effects of Antioxidants on NF-KappaB Activation in Bovine Carotid Artery Endothelial Cells. AB - NF-Kappa B is one of important transcription factor, which plays an important role in many immune and inflammatory responses. A large variety of stimulus can induce the degradation of IKappa B-alpha, an inhibitor of NF-KappaB, leading to the activation and translocation of NF-KappaB into nuclei. In the present study, the effects of antioxidants on TNF-alpha-induced NF-KappaB activation were tested by using cultured bovine carotid artery endothelial cells. The degradation of IKappaB-alpha were checked by Western blot, and the nuclear translocation of NF KappaB were tested by immunofluorescence. Our results showed that pyrrolidine dithio-carbamate (PDTC) completely inhibited IKappaB-alpha degradation and nuclear translocation of NF-KappaB induced by TNF-alpha, and other antioxidants also exhibited inhibitory activities on TNF-alpha-induced I B-alpha degradation. PMID- 12075420 TI - Adenovirus-Mediated th Gene Therapy of Parkinson's Disease in a Experimental Rat Model. AB - A recombinant adenovirus (AdCMV th) encoding tyrosine hydroxylase (TH) gene with CMV promoter was constructed and propagated. Southern blot analyses was used to identify positive plaques. Virus titer was about 1.4x10(14) pfu/L as determined by plaque forming assay. In glial cells infected with AdCMV th, the TH expression was demonstrated by immunohistochemical staining and HPLC-ECD. 678.8 ng DA was detected in the extract of 1x10(6) AdCMV th infected glial cells, but no detectable DA was found in AdCMVLacZ-infected glial cells. Injection of AdCMV th (1x10(7) pfu/rat) into the striatum of PD rats significantly improved the apomorphine-induced rotation movement(approximately 60%). The improvement in rotation movement remained up to 5 months after injection, and TH expression positive cells were found in the vicinity of injection. These results indicate that adenovirus may be a useful carrier for in vivo gene therapy in the PD patients. PMID- 12075421 TI - Purification and Primary Structure of a Novel Peptide from the Chinese Scorpion Buthus martensi Karsch. AB - To purify and characterize peptides from the venom of Chinese scorpion Buthus martensi Karsch, the purification was carried out by gel-filtration, ion exchange and reversed phase HPLC techniques. The purified peptide was reduced by dithioerythritol (DTT), S-alkylated with iodoacetic acid, and subjected to enzymatic cleavages (TPCK-trypsin). The purified fragments from enzymatic cleavage of the peptide were separated by C(18)HPLC, then submitted to the ESI MS, and Edman degradation for amino acid sequence determination. The mixture was also subjected to tandem mass (MS-MS) analysis. As a result, a novel peptide, named BmK4112, was obtained, with the primary structure being TPYPV NCKTD RDCVM CGLGI SCKNG YCTGQ C, and having three disulfide bonds. PMID- 12075422 TI - Upregulation of bax Gene Expression Promotes Paclitaxel-induced Apoptosis in Esophageal Carcinoma Cells. AB - An inducible mammalian expression vector of bax gene was constructed and the control ability of metallothionein II promoter in esophageal carcinoma cell line was systematically identified with luciferase report gene. After the transfection of it into human esophageal carcinoma cell line Eca109, Bax protein expression was analyzed by immunolcytochemical method. Paclitaxel-induced apoptosis was determined by TUNEL assay, DNA ladder assay and flow cytometry. Results showed that 140 &mgr;mol/L ZnSO(4) for 12 h is optimal for induction of bax gene expression. Under these conditions, a clonal transfectant X1097(#), expressing bax gene effectively, was obtained. It was found that, X1097(#) had higher apoptotic rate and was more sensible than Eca109. These results implied that Bax protein may play an important role in paclitaxel-induced apoptosis. Therefore, bax protein may be promising as an helping drug to improve therapeutic effects of paclitaxel. PMID- 12075423 TI - Cloning and Analysis of Intron 3 of Human Telomerase Reverse Transcriptase (hTERT) Gene. AB - Total RNA was extracted from telomerase-positive cell line SPC-A, and reverse transcribed to cDNA. The long template PCR was performed using this cDNA as template and the hTERT cDNA specific oligonucleotides as primers. A long fragment of about 2.2 kb was produced as well as a short fragment of about 150 bp. The long fragment was purified from the gel, cloned into T-easy vector and sequenced from two directions. The sequencing result and homologous comparison indicated that the fragment contained the intron 3 of hTERT gene. Further assay showed that the precursor of this fragment is premature mRNA (pre-mRNA) of hTERT gene and the copy number varied among different cell lines, as verified in this study. These results suggest the feasibility of cloning the intron of eucaryotic gene by the combination of reverse transcription and long template PCR system. And the different splicing efficiency of the intron 3 from the hTERT pre-mRNA was also implied from this assay. PMID- 12075424 TI - Cloning of Human Myelin Protein Zero-like Genes by Bioinformatics Strategy. AB - To clone novel myelin protein related genes, two human ESTs, which shared significant similarity with the human myelin protein zero gene, were found by the comparison of homologue between the cDNA coding region sequences of MPZ gene and the EST database of NCBI. An 801 bp EST contig was assembled, which was 100% identical with a 128 kb genomic sequence, mapped to 1q24. A 435 bp open reading frame (ORF) within the 801 bp contig was shown by computer analysis. Two primers designed according to the sequence of the contig, were coupled with the primers(lambdagt10-5 and gt10-5) on the sequences flanking cloning site of the cDNA library vector to amplify the cDNA library sequences by nested PCR. New primers, designed based on novel cDNA sequences, were used for the PCR amplification with lambdagt10-5 and gt10-5 in the same way as above. Finally, the human myelin protein zero like gene isoform I and II (MPZL1a, MPZL1b GenBank AF095727, AF092424) were cloned. Comparison of gene and protein structures between MPZL1 and MPZ revealed that MPZL1 is the second member of MPZ family. Mutation analysis of MPZL1 gene was performed in 24 Charcot-Marie-Tooth disease (CMT) families and 26 nonsyndrome deafness families, but no mutation was found. PMID- 12075425 TI - Structure of a New Glucan from Cynanchum panilatum (Bunge) Kitagawa. AB - A polysaccharide was purified from a traditional Chinese drug, "Xu-Chang-Qing" (Cynanchum panilatum (Bunge) Kitagawa), and its molecular weight was estimated to be 1.5x10(4) by HPLC method. Its specific rotation [alpha](D) was 151.4 deg; (0.96, H(2)O). Sugar composition analysis indicated this polysaccharide was composed only of glucose. Methylation analysis, partially hydrolysis, acetolysis, IR and NMR data showed that its main chain was composed of alpha-D 1,4 linked glucosyl residues and its side chains were composed of alpha-D 1,4 and alpha-D 1,6 linked glucosyl residues. There was one branch located at O-6 of the glucosyl residue in every repeating unit containing five-glucosyl residues. PMID- 12075426 TI - Development and Preliminary Application of a Peptide Mass Fingerprinting Technique in Proteome Research. AB - Proteome means the total proteins expressed by the genome in a cell or tissue. Two-dimensional electrophoresis (2-DE) is now the most powerful separating technique and the key separation method used in proteome. Peptide mass fingerprinting (PMF) is becoming a widely used and vastly developed technique for protein identification in 2-D gels. In this research, a systematic method to identify the proteins in polyacrylamide gels by PMF was developed. Proteins were digested in-gel by enzyme and the masses of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The data obtained from PMF were used in protein database search and the protein identification. The proteins from human lung cancer cells isolated by 2-DE were subjected to identification by the PMF method developed in this work. Three spots of proteins in gel were identified as G3P2_HUMAN, UBL1_ HUMAN and TPIS_HUMAN. PMID- 12075427 TI - Kinetics of Ionizing Irradiation-induced DNA Strand Breaks. AB - By measuring the dose effect of ionizing irradiation-induced single- and double strand breaks (SSB and DSB) of pBR322, the responses of the yields of SSB and DSB to DNA concentrations were obtained. It was found that, in the presence of mannitol, the reciprocals of G(SSB) and G(alphaDSB) had a linear relationship to the reciprocal of DNA concentration so that the competition reaction of DNA and mannitol to.OH radicals could be described by a second-order kinetics. Moreover, the rate coefficients and the efficiencies of the.OH radical inducing SSB and alphaDSB were deduced. PMID- 12075428 TI - Induction of Apoptosis in Tumor Cells by a Recombinant Adenovirus with bak Gene. AB - Apoptosis, a genetically encoded process of cellular suicide, comprises an intrinsic cellular defence against tumorigenesis. bak(for bcl-2 homologous antagonist/killer) is a new apoptosis-inducing gene found recently, which is identified in all tissues examined. Initial work of Farrow, Chittenden and Klefer indicated that bak plays an important role in apoptosis. Successive work of others suggests inactivity of Bak always accompanies the development of tumor cells. In this work, a shuttle vector harboring bak gene, pCA13-bak, was constructed and used to cotransfect into the cell line 293 together with a recombinant plasmid, pBHG11, containing most of adenovirus genome. A mutant adenovirus(delete E1, E3 region)carring bak, Ad-bak, was obtained, and purified. Dot blot identified the construction of recombinant virus was successful. HeLa cells were infected by this virus. Some manners were used to identify the changes of HeLa cells including Annexin V-FITC staining which made the reversed PS observed directly, microscope observation and others. The characteristic morphology of apoptotic cells was due to Ad-bak. The experiment suggests that bak is a strong apoptosis-inducing gene, and a potential gene which could be used in gene therapy on tumor. PMID- 12075429 TI - Identification of Protein Spots in Silver-stained Two-dimensional Gels by MALDI TOF Mass Peptide Map Analysis. AB - Protein spots in silver-stained two dimensional gels were analyzed and identified by employing an improved procedure of mass spectrometric peptide mapping, including i) In-gel reduction, alkylation and enzymatic digestion ii) Extraction and desalting by using a pipette tip containing a small C18 micro-column (ZipTip(TM)) iii) Direct MALDI-TOF mass analysis and protein database searching. The results demonstrate that single silver-stained protein spots in a 2-DE-gel could be identified rapidly by this procedure and the use of the ZipTip(TM) pipette tip could increase evidently the sensitivity of the MALDI-TOF analysis. By using this the procedure, 10 protein spots in a silver-stained gel of 2-DE of crude venom of the spider S.Huwena were analyzed and identified. PMID- 12075430 TI - Cloning of Human IgG Fc cDNA and Expression of Whole Human Anti-HBsAg Antibody in CHO Cells. AB - Messenger RNA was extracted from human peripheral lymphocytes and first strand cDNA was prepared by reverse-transciption. The cDNA of Fc fragment of human IgG1 was then obtained by PCR and was cloned into the pGEM T-vector. The DNA sequences encoding signal peptides of both light and heavy chains were synthesized and cloned respectively. For construction of the light chain expression plasmid, the light chain signal sequence was linked with the light chain variable and constant regions (VL-CL) which had been cloned previously by screening of phage display libraries with HBsAg. The resulting full-lenth light chain sequence was then inserted into pcDNA3.1, a mammalian expression vector. For construction of the heavy chain expression plasmid, the heavy chain signal sequence, the variable region, the first constant region (VH-CH1, cloned previously by screening of phage display libraries with HBsAg) and Fc fragment sequence were ligated to form a full-length heavy chain ORF, which was then cloned into another mammalian expression vector, pCI-DHFR1. CHO(dhfr(-)) cells were cotransfected with the above light and heavy chain expression plasmids, and cell clones expressing human anti-HBsAg antibodies were selected by G418 and methotrexate (MTX). The recombinant human antibodies were purified with protein L affinity chromatography from the cell culture medium. As human serum IgG, the recombinant IgG exhibited only one band with a molecular weight of more than 100 kD in non-reducing SDS PAGE in reducing SDS-PAGE, however, it turned out to be two bands of approximately 50 kD and 25 kD respectively. Western-blot analysis demonstrated that the whole IgG in the non-reducing SDS-PAGE, and the heavy chain in the reducing SDS-PAGE both reacted with goat anti-human Fc antiserum. PMID- 12075431 TI - Interleukin 18 High Level Expression in E.coli Purification and Renaturation of the Recombinant Protein. AB - Using the total RNA extracted from mitogen-stimulated human peripheral blood mononuclear cells (PBMC) as template, the cDNA of interleukin 18 was amplified by RT-PCR. The cDNA was subsequently cloned into the expression vector pJW2 and sequenced. The recombinant human IL-18 (rhIL-18) was expressed efficiently in inclusion bodies in E.coli with the yield accounting for 20% total bacteria proteins. The inclusion bodies were washed with 2 mol/L urea and rhIL-18 was further purified using Sephadex G-100 column chromatography in 8 mol/L urea. After purification, the purity of rhIL-18 was greater than 90% as judged by SDS PAGE. The purified rhIL-18 showed significant and dose-dependent IFN-gamma inducing activity in human PBMC, in the presence of 0.5 mg/L Con A. PMID- 12075432 TI - Detection of Lis1 Gene Frame Shift Mutation in Human Hepatocarcinoma. AB - GST fusion protein expression system combined with protein truncation test(PTT) protocol was used to detect gene frame shift mutation. The RT-PCR products of Lis1 genes from hepatocarcinoma samples were respectively cloned into a GST fusion protein expression vector pGEX-1, then expressed in E.coli. The results showed a truncated 33 kD fusion protein in SDS-PAGE, although the full-translated product of Lis1 gene should be of 71 kD. Sequencing revealed insertion of an A residue, causing the premature termination of translation, between the 163th and 164th nucleotide of Lis1 gene. This improved PTT assay was proved to be a fast and effective way in detecting gene frame shift mutation. PMID- 12075433 TI - Phosphatidylinositol(PI)-3 Kinase Contributes to the Activation of Stat3 in M1 Leukemia Cells Induced by IL-6. AB - Wortmannin, a specific inhibitor of PI-3 kinase, antagonized the inhibitory effect of IL-6 in M1 acute myeloid leukemia cells. Electrophoretic mobility shift assays revealed that wortmannin selectively reduced the IL-6-induced activation of Stat3, while Stat3 plays a central role in IL-6-induced growth-arrest and terminal differentiation of M1 cells. These results indicated that PI-3 kinase was involved in the signal transduction of IL-6 and contributed to the activation of Stat3. PMID- 12075434 TI - Comparison of the Catalytic Domains of Collagenase-1 and Stromelysin-1. AB - The catalytic domains of two matrix metalloproteinases--collagenase-1 and stromelysin-1 have been studied by means of fluorescence spectroscopy and high hydrostatic pressure. The hydrophobic fluorescence probe ANS could bind to stromelysin-1, with a dissociation constant of 26.3 &mgr;mol/L, but could not bind to collagenase-1, indicating that there exists a hydrophobic site on the surface of stromelysin-1. Further study suggests that the hydrophobic site may not be the catalytic site. The biological activity of catalytic domains of collagenase-1 and stromelysin-1 showed obvious difference under high pressure the activity of collagenase-1 increased with elevating pressure, with an activation volume of D18.9 ml/mol however, the activity of stromelysin-1 did not change under high pressure. The results indicate that there are some obvious differences between the catalytic domain conformations of these two enzymes, though the crystal analysis indicated that they were very similar as reported before. PMID- 12075435 TI - NGF-Tf Conjugate Prevents Degeneration of Substantia Nigra Neurons in a Mouse Model of Parkinson's Disease. AB - Nerve growth factor(NGF) was purified from Naja naja atra snake venom and conjugated to transferrin(Tf). This conjugate was intravenously injected into a mouse model of Parkinson's disease (PD). Both immunohistochemical staining and pathological detection showed that the NGF-Tf conjugate could prevent the loss of tyrosine hydroxylase-immunoreactive neurons located in substantia nigra and the cell counts of NGF group were 2 330.0+/-260.3, and those of the MPTP group and the control group were 797.0+/-121.4 and 2 381.0+/-158.0, respectively. In addition, electron microscopic examination revealed significant protection against demyelination and vacuolation in subtantia nigra neurons in contrast to the control group. The i.v. injected NGF-Tf conjugate also reversed the neurodegenerative changes such as karyopyknosis, chromatolysis and intracytoplasmic inclusion in diseased neurons. PMID- 12075436 TI - Deinococcus radiodurans CatB Gene Cloning and Expression in Escherichia coli. AB - A 1 611 bp length complete coding sequence of the catalase (Cat) was obtained through bioimformatical analysis of the database of D.radiodurans genome, and then was amplified from D. radiodurans genomic DNA by polymerase chain reaction. The amplified gene was cloned into pKK223-3 vector and transformed into E.coli UM2, a Cat-deficient mutant. Staining of non-denaturing polyacrylamide gels for Cat activity demonstrated that pKK223-3 insert encoded a Cat that co-migrated with the CatB found in D.radiodurans cell lysates. Expression of CatB gene from D.radiodurans in E.coli UM2 restored the resistance to H(2)O(2) at low concentrations. PMID- 12075437 TI - Molecular Design and Bio-activity Analysis of a t-PA Derivative (t-PA S165W). AB - In order to obtain tissue-type plasminogen activator (t-PA) mutant with enhanced affinity for fibrin, redesigning t-PA strategies were proposed by computer molecular designing technology, and the re-designed t-PA derivative, t-PA S165W, was expressed in CHO cells. The bio-activities and affinity for fibrin of the expression product were tested. No apparent changes were observed between t-PA S165W and wild type t-PA. Therefore, single site mutation of the t-PA could not lead to enhanced affinity for fibrin of t-PA. PMID- 12075438 TI - Molecular Cloning and Sequence Analysis of cDNA Encoding Acutolysin C, a Hemorrhagic Metalloproteinase, from Agkistrodon acutus. AB - A full-length cDNA of 1 650 bp was amplified from the snake venom gland cDNA library of Agkistrodon acutus. Analysis of the nucleotide sequence indicated that the amplified cDNA contained a complete open reading frame encoding 417 amino acid residues including signal peptide sequence, zymogen sequence and proteinase domain. The zymogen sequence contained CGVT motif that was highly conserved in almost all venom metalloproteinases. The metalloproteinase domain contained a conserved signature zinc-binding motif HEXXHXXGXXH in the catalytic region and the CIM turn. It shares high similarity with the sequence of acutolysin C deduced from crystallographic data, and with other class P-I snake venom hemorrhagic toxins. PMID- 12075439 TI - Cloning and Expression of Human CD20 Gene on NIH-3T3 Cell Membrane. AB - CD20 is a specific antigen expressed on normal and neoplastic B cells exclusively. Recent researches showed that in B cell leukemia, CD20 was over expressed. Therefore monoclonal antibody (McAb) to CD20 may be of clinical value in diagnosis and treatment of some leukemias and lymphomas. In this study, the full length gene of CD20 cDNA were cloned from total RNA of Raji cells, inserted into an eukaryotic expression vector pcDNA3.1, forming a recombinant plasmid pcDNA3.1/CD20. NIH-3T3 cells were transfected with pcDNA3.1/CD20 and selected with G418 for the transfected cells. Alkaline phosphatase against alkaline phosphatase assay(APAAP) experiments showed that the selected cells could express the human CD20 onto its surface. Balb/c mice were immunized with CD20( ) NIH-3T3 cells once three weeks for 3 shuts. Indirect immunofluorescence experiments were done with the Raji cells and the serum of the immunized mice, and the results showed that the spleen of the immunized mice could be used to prepare the McAb to CD20. PMID- 12075440 TI - Lectins and Toxins. AB - Although lectins, in general, are not very toxic, there are some relationships between lectins and toxins. BEA-type toxins can be divided into two main parts. One shows sugar-binding activity responsible for targeting. Another part, which possesses different enzymatic activity, determines the toxicity. The lectins isolated from Trichosanthes kirilowii share some properties with BEA-type toxin, e.g. ricin. Some toxins can perforate cell membrane, forming an ion channel. A lectin from Pinellia turnata may belong to this kind of toxin. Several lectins were isolated from snake venom, sharing homologous amino acid sequences with other components in venom, while a receptor of well known toxin in venom, phospholipase A(2), contains lectin-like domain. In the venom of Trimeresurus stejnegeri exists a C-type animal lectin. The conclusion is that both lectins and toxins may be involved in the defense mechanism of organisms. PMID- 12075441 TI - Progress in the Research of Plasminogen Activator Inhibitor Type-2. AB - Plasminogen activator inhibitor type-2 (PAI-2) inhibits urokinase type plasminogen activator (u-PA) most specifically and high efficiently, following the mechanism of serine proteinase inhibitor (serpin) superfamily. PAI-2 plays a very important role in vivo there are, however, two conflicting views on the role of PAI-2 in cancer. Tissue-type plasminogen activator, vitronectin, transglutaminases, fibrin and many other molecules can interact with PAI-2. Regulation factors of PAI-2 gene expression includes many regulatory elements in the promoter region, a number of agonists and conditions of organism, etc., showing that PAI-2 gene is regulated by both positive and negative mechanisms. PMID- 12075442 TI - The Application of a Novel Lytic System to the Recovery of Recombinant Proteins in E.coli. AB - In order to efficiently recover recombinant proteins, a temperature-sensitive lytic system was constructed on the basis of the feature that T4 lysozyme disrupts the bacteria through cutting specific bond in the peptidoglycan layer of cell wall. This system was evaluated by constructing and introducing a low copy plasmid pSC-lys (pSC101 replication origin) into E.coli. The plasmid contained a temperature sensitive T4 lysozyme (LYS(ts)) gene under the control of three tandem tac promoters and the LacI repressor, which is compatible with other plasmids carrying pMB1, ColE1 replication origins, etc. Under the optimum lysis conditions, 2--5 fold condensed cultures resuspended in buffer A, beta galactosidase, recombinant chaperone GroEL and ZZ-fusion salmon hexamic calcitonin (Cal6) in E.coli were released simply, rapidly, and quantitatively, as co-expressed with LYS(ts). The two tested recombinant proteins maintained their significant productions. Instead of other cumbersome lysising methods, this novel lytic system will be useful in recovery of recombinant proteins for further purification in the field of biotechnology. PMID- 12075443 TI - Expression of Human Flt3 Ligand in Pichia pastoris and Its Biological Characteristics. AB - Flt3 ligand(FL) is a cytokine that stimulates the proliferation and differentiation of hematopoietic stem cells/progenitors. In order to obtain high level of recombinant human soluble FL(rhFL) production, an artificial gene for rhFL was synthesized by using favored genetic codons of the yeast Pichia pastoris. Then the gene was cloned into the vector pPICzalphaA and the resulting construct was introduced into Pichia pastoris for expression. It was found that it was possible to obtain biologically active rhFL with a yield of over 30 mg/L of yeast culture. The rhFL stimulated colony formation from cord blood. rhFL, SCF, GM-CSF and IL-3 are the most promising combination for the in vitro expansion of stem/progenitor cells. It was also showed that rhFL increased the induction of dendritic cells from cord blood in combination with GM-CSF, TNFalpha and IL-4. Interestingly, rhFL stimulated the growth of an endothelial cell line this effect has not been reported before. PMID- 12075444 TI - Preferential Location of MITEs in Rice Genome. AB - To observe the association between MITEs and genes in details, 82 kb of the rice genomic DNA were sequenced, and 10 genes were identified by similarity search. It was found that miniature inverted-repeat transposable elements (MITEs) were located upstream of the genes identified without exception, regardless of their directions of transcription. One MITE was found to be located between two 5' regions of the genes with opposite orientation of transcription, indicating that it was shared by them. The positional patterns of MITEs may therefore serve as an aid for preliminary screening of genes. PMID- 12075445 TI - Transcription and Translation of Sendai Virus Hemagglutinin-neuraminidase (HN) in Mammalian Cells. AB - Sendai virus hemagglutinin-neuraminidase was expressed in COS-7 cells. LLCMK(2) cells were transfected with expression plasmids harboring HN, namely pJH3, pJH4 or pJH7, and were passaged conti-nuously under the presence of hygromycin or puromycin to get antibiotic-resistant clones, which had HN gene integrated into genomes. S1 nuclease assay indicated that large quantity of HN mRNA was transcribed but indirect-immunofluorescence and immunoprecipitation demonstrated that there was little HN protein expressed on cell surface and inside cells, in contrast with the Sendai virus persistent-infected LLCMK(2) cells that expressed largely HN protein. Therefore, not every transcription resulted in translation. It also suggests that some virus factors may positively regulate transcription and translation of HN mRNA. PMID- 12075446 TI - Purification of Recombinant GM-CSF/IL-3 Fusion Protein. AB - As a new artificial haemopoietic growth factor, GM-CSF/IL-3 fusion protein appears very promi-sing to be developed as a drug in the treatment of many diseases including cancer. The purification process of the recombinant GM-CSF/IL 3 fusion protein was studied. Similar to majority of the eukaryotic proteins, GM CSF/IL-3 was expressed as inclusion bodies in E.coli. A series of purification steps, including cell breakage, inclusion body washing, inclusion body solubilization, protein renaturation and ion-exchange chromatography, have been set up to purify the recombinant fusion protein in an active form. Experimental results showed that the inclusion body solubility increased with increasing urea concentration. During the protein renaturation process, dialysis method was chosen to remove the denaturant urea. Stepwise decrease of urea concentration in the buffer could effectively improve the protein renaturation efficiency by reducing protein aggregation. At the same time, reduced and oxidized glutathionine were added to optimize the correct disulfide bonds formation. The recombinant protein was then purified by DEAE ion-exchange chromatography. The final protein recovery was over 30%. SDS-PAGE and reverse HPLC analysis revealed over 95% purity of the final purified recombinant protein. Therefore, a laboratory-scale purification procedure of the recombinant GM-CSF/IL-3 fusion protein was basically well established. PMID- 12075447 TI - Expression of Smad4 Gene in Pichia pastoris and Identification of the Protein. AB - Smad4 is a novel tumor suppressor gene which is mutated or deleted in about 50% of pancreatic carcinoma and 30% of colon cancer. SMAD4 was expressed in Pichia pastoris and was characterized. The molecular weight of the expression product was shown to be about 67 kD, and 13 amino acids in N terminal were determined and were identical with the putative sequence from Smad4 cDNA, and it was able to specifically react with the antibody against SMAD4. PMID- 12075448 TI - A Structure-function Analysis of Human GDNF. AB - The glial cell line-derived neurotrophic factor(GDNF) plays a very important role in the regeneration of the nervous system. Based on the results of the X-ray structure analysis of rat GDNF, human GDNF gene was modified with deletion and insertion mutagenesis by using PCR methods. The various mutants were all highly expressed in E.coli. The recombinant proteins were purified and their survival promoting activities were determined by using cultures of the spinal cord neurons of embryonic mouse. The results showed that the "cystine knot motif" was critical for the maintenance of GDNF structure the alpha-helix, finger 1 and finger 2 region were critical for GDNF neurotrophic activity and the N-terminus of human GDNF was not essential for its biological functions. PMID- 12075449 TI - Oxidative Modification Reduces HDL-induced Cholesterol Efflux from Cultured Human Arterial Smooth Muscle Cells. AB - To investigate the effects of oxidative modification of high density lipoprotein (HDL) on HDL-mediated cholesterol efflux from human arterial smooth muscle cells(SMC), native HDL (N-HDL) and oxidized HDL(Ox-HDL) induced by copper or hypochlorite were incubated with cultured SMC preloaded with (3)H-cholesterol for 2--24 h. The radioactivities of (3)H-cholesterol of the medium and SMC were measured respectively, and the cholesterol efflux rate (CER) of HDL and Ox-HDL were determined. The result showed that the CER of N-HDL was 43.1% after 24 h incubation, while the CER of Ox-HDL induced by copper or hypochlorite were 28.9% and 23.5% respectively, reduced by 30.0% and 43.1% compared with N-HDL. The speed of 3H-cholesterol efflux of Ox-HDL from SMC were decreased remarkably. Inhibition of oxidation with antioxidants butylated hydroxytoluene(BHT), vitamin C or vitamin E recovered mostly the ability of HDL to stimulate cholesterol efflux. The results suggest that oxidation of HDL may attenuate its effect on cholesterol efflux from arterial SMC, and vitamin C and vitamin E supplementation may protect artery from atherogenesis. PMID- 12075450 TI - The Sequence Analysis and in Vitro Cleavage by Ribozymes of a Putative Early and Late Transcription Gene of the Prawn Baculovirus. AB - Since 1993 there has been outbreak of an acute lethal disease of the cultivated fleshy prawns (Penaeus chinesis, Osbeck) in China. After a short period of intensive studies we reported firstly the presence of a non-occluded baculovirus in the tissue of diseased prawns. The virus was also named as white spot syndrome baculovirus by some authors. Here, it is reported that, based on sequence analysis, an early and late transcription gene(1 197bp gene)of baculovirus was identified. Two hammerhead ribozymes RZ1 and RZ2 targeting the 54-56 bp and 314 316 bp of the 1 197 bp gene were designed, and in vitro cleavage experiments showed that under proper conditions the target gene could be completely cleaved by those ribozymes together or separately. PMID- 12075451 TI - Signal Transduction in TNF-alpha-induced c-jun Gene Expression. AB - Our previous studies demonstrated that p38 mitogen-activated protein (MAP) kinase regulated the c-jun protein expression through phosphorylation of transcription factors of myocyte enhancer factors 2 (MEF2) family. There was a MEF2 binding site in the promoter of c-jun gene. Members of the MEF2 family of trans-cription factors bound as homo- and heterodimers to this MEF2 binding site. Here the potential role of the p38 and BMK1 MAP kinases in the regulation of c-jun expression induced by TNF-alpha was examined. It was shown that p38 MAP kinase up regulated the transcription activity of MEF2A, while BMK1 MAP kinase up-regulated not only the transcription activity of MEF2A, but also MEF2D. The p38 and BMK1 MAP kinases had coordinated effect on the regulation of c-jun transcription. TNF alpha induced the formation of MEF2A/MEF2D hete-rodimer. Over-expression of homodimer of MEF2 proteins inhibited c-jun transcription induced by TNF-alpha, while over-expression of heterodimer MEF2A/MEF2D enhanced c-jun transcription induced by TNF-alpha. Phosphorylation of MEF2A and MEF2D by p38 and BMK1 respectively appeared very important in TNF-alpha induced MEF2A/MEF2D heterodimer formation to enhance c-jun gene expression. PMID- 12075452 TI - Retention and Thermodynamic Properties of Three Insulin Variants on the Reversed phase Liquid Chromatography. AB - The retention and thermodynamic behaviour of three insulin variants, bovine, human and porcine insulins, in reversed-phase high-performance liquid chromatography were studied over a range of temperatures between 10 and 65 degrees and a range of methanol concentrations between 53 and 59%(v/v). The results demonstrated that the slight difference of three insulins in the amino acid sequence could be resolved significantly in C(8)-hydrophobic ligand. Values for the relative changes in enthalpy (deltaH deg;/R) and entropy (deltaS deg;*) associated with the interaction process were also determined. These values of thermodynamic parameters would provide further insight into the factors involved in the stabilization of protein conformation and the mechanism of the interaction of peptides with hydrophobic surfaces. The experimental results also demonstrated that the determination of thermodynamic parameters of interactions between peptides and hydrophobic surfaces would provide an alternative approach for the investigation of mechanisms of protein folding and of interaction between proteins. PMID- 12075453 TI - Rapid Construction of Full-length cDNA Clones of Tobacco Mosaic Virus and the Infectivity Assay of Its in Vitro Transcript. AB - The full-length cDNA of tobacco mosaic virus faba bean isolate (TMV-B) was amplified with RT-PCR in which T7 promoter sequence was added in the 5' terminus of its upstream primer, so that full-length cDNA was put directly under the control of a T7 promoter. The cDNA was cloned into plasmid pT7Blue and linear DNA was got by digesting the recombinant with KpnI or KpnI and PstI. Using these linear DNA and full-length PCR product as templates, respectively, their in vitro transcripts were inoculated to Nicotiana tabacum and Chenopodium amaranticolor. All of the transcripts had infectivity and produced symptoms similar to that of wild TMV. It was found that transcripts of the full-length PCR product had higher infectious efficiency than those of linear DNA. PMID- 12075454 TI - A Chimera Polypeptide with Active Sites of HWTX-I and AAI. AB - The functional amino acid sequence and its neighbouring fragments in the molecule of Amaranth alpha-amylase inhibitor isolated from seeds of the Mexican crop plant Amaranthus hypochondriacus were transferred, by solid phase chemical synthesis, into N-terminal region of the huwentoxin-I(HWTX-I). The synthetic chimera polypeptide was confirmed by Edman degradation and MALDI-TOF mass spectroscopy. The formation of three disulfide bonds and special conformation of the synthetic chimera was induced by the addition of glutathione. Renatured chimera polypeptide was purified by ion-exchange and reversed phase HPLC. The results showed that the engineered chimera polypeptide exerted obvious inhibitory activity to alpha amylase from digestive tube of the roach (Periplaneta Americana) at pH 5.5 with the concentration of 9.5x10(-5) mol/L, and also exerted 36% of the neurotoxic activity of the natural huwentoxin-I as shown by the experiments of blockage of the neuromuscular transmission of isolated mouse phrenic nerve-diaphragm preparations. The experiments demonstrated that the structural motif of HWTX-I is well promising for protein engineering, and the solid-phase peptide synthesis is adequate rapid for the engineering of artificially designed small proteins. PMID- 12075455 TI - Infectious Bursal Disease Virus Structural Protein VP2 Expressed by a Baculovirus Recombinant in Bombyx mori. AB - VP2 cDNA gene of the infectious bursal disease virus HZ96 strain, encoding a major host-protective antigen, was cloned into baculovirus transfer vector pBacPAK8, resulting in a recombinant transfer vector pBacPAK-VP2. The vector pBacPAK-VP2 and linearized DNA of modified baculovirus Bm-BacPAK6 were co transfected into the cultured Bombyx mori (Bm) N cells, in which homologous recombination occurred. Then, baculovirus recombinants were screened out. The Bm cells and Bm larvae were infected with the baculovirus recombinant that can expresse VP2, and Bm N cells and haemolymph of Bm larvae were collected for assays. The results of ELISA and Western immunoblotting assays demonstrated that VP2 was expressed in the cultured Bm cells and the Bm larvae. PMID- 12075456 TI - DNA Microarray Chips Made on Surface of Ceramic Slides. AB - As a novel technology, DNA microarray chips are being used in gene detections. To make DNA chips capable of multiple probing, ceramic slides, instead of silicon wafer or glass slides, were used as the substrate upon which DNA microarray are deposited. Our study indicates that the ceramic DNA microarray chips can be applied to multiple hybridization and detection procedures, showing high specificity. The advantages in using ceramic substrates are discussed. PMID- 12075457 TI - High Expression of Human Persephin in Insect Cells. AB - The human persephin (PSP) was expressed in Tn-5B1-4 insect cells using the Bac to Bac baculovirus expression system. The expressed product amounted for 20% of total cellular soluble proteins. The expressed product was purified by Ni(2+) affinity chromatography and the activity assays showed that it could significantly prolong the survival of spinal cord neurons. PMID- 12075458 TI - Expression of Human Epiregulin in E.coli. AB - Human epiregulin cDNA was amplified from the lung cancer cell line A549 using RT PCR. After adding 6 His codon to its 3' end, it was cloned into a high efficient secretive Escherichia coli system with alkaline phosphatase promoter(phoA promoter)constructed in our lab and induced for expression. The product was purified one-step by Ni-NTA column. Amino acid sequence analysis revealed the identity of our product with that previously reported. The product showed strong proliferative effect on fibroblast cell line Balb/c3T3 and growth inhibitory effect on epithelial carcinoma cell line A431. PMID- 12075459 TI - CEK2, a Novel MAPK from Candida albicans Complement the Mating Defect of fus3/kss1 Mutant. AB - A novel MAPK(mitogen-activated protein kinase) gene, CEK2(Candida albicans extracellular signal-regulated kinase 2), was isolated by screening the Candida albicans library based on oligonucleotide probe hybridization and degenerated PCR. The CEK2 gene is 1 119 bp in length, and coding for a 373 aa protein. The CEK2 shares 56% homology with CEK1 from Candida albicans, 55 % homology with FUS3 from S. cerevisiae. From the deduced amino acid sequence, the Cek2 protein contains a conserved ATP binding site and a Ser/Thr kinase activity signal and a conserved TEY sequence was located at L(12) region. In vitro kinase activity assay showed Cek2 could phosphorylate MBP(myelin bovine protein) but not histone H1. CEK2 gene could complement the fus3/kss1 mutant and underwent mating signal induction by a tester strain, but CEK1 gene could not complement with the fus3/kss1 mutant. CEK2 is therefore a FUS3 homolog in Candida albicans. PMID- 12075460 TI - Molecular Cloning of MAPK Gene Family Using Degenerate PCR. AB - MAPK(Mitogen-activated protein kinase) pathways play an important role in morphorgenesis of Candida albicans. According to the conserved amino acid sequence of the known MAPKs, two degenerate primers attaching to subdomain VIB and IX were designed to screen novel MAPKs in C. albicans. The PCR was performed under non-strigent conditons. 100 PCR fragments were sequenced and among 25 novel gene fragments, two novel MAPK gene fragments were obtained. Using these two PCR fragments as probes to screen a Candida albicans genomic DNA library, two novel MAPK genes designated CSK1(Candida albicans sporulation-related MAP kinase1) and CEK2(Candida albicans extracellular signal-regulated kinase 2) were cloned. These two genes share high similarity with three cloned MAPK genes CEK1 and MKC1 and CaHOG1. The CSK1 gene is 1 193 bp in length, containing a 92 bp intron, coding for a 367 aa protein. The CSK1 shares highest similarity with SMK1, with homology 55.3% in nucleotide sequence and 50% in amino acid sequence. SMK1 encodes a MAPK involved in the sporulation pathway in S. cerevisiae. In vitro kinase activity assay showed that the Csk1 kinase exhibited a phosphorylation ability when using MBP as a substrate but not histone H1. PMID- 12075461 TI - In Vitro Amidating Processing of Products Expressed by Gene Engineering. AB - To set up an in vitro amidating system, a recombinant human calcitonin with a glycine at C termial (mhCT-Gly) was used as the amidating substrate of recombinant rat peptidylglycine alpha-amidating monooxygenase (rPAM). First, the mhCT-Gly gene was synthesized and cloned into a fusion expression vector to get an expression plasmid pGEXCT. The GST-fused mhCT-Gly was highly expressed in E.coli BL21(DE3) harboring the pGEXCT, and was purified rapidly by affinity chromatography. Second, using the method of ultrafiltration, the rPAM was prepared from the supernatant of cultured transfectant CHO cells which express rPAM stably. Finally, the in vitro amidating experiments were carried out using GST-mhCT-Gly as substrate and the prepared rPAM. The results of dot blot with the specific antibody and of mass spectrum assay indicated that amidating product GST hCT-NH(2) could be easily detected. This study provides a useful method for the amidation of recombinant products in vitro. PMID- 12075462 TI - [A brief introduction to the secretion mechanism in immunocytes]. AB - Exocytosis is a vital function of many cell types including neuron, endocrine cell and immunocyte. Secretion in immunocytes involves a complex process of signal transduction, in which many factors still remain unknown. In the last 10 years, this area has become an international hot spot of investigation, resulting in many break-through progresses. This progress was made possible by combined efforts in molecular biology, cell biology and biophysics. This review focuses on notable new knowledge and some new techniques in functional study of secretion in immunocytes. PMID- 12075463 TI - Serotonin excites arcuate neurons directly but inhibits them through intercalated GABAergic neurons. AB - Effects of serotonin (5-HT) on spontaneous discharges of single hypothalamic arcuate neurons were observed in rat brain slices by extracellular recordings. The results showed that (1) of 149 neurons selected randomly and tested for 5-HT application, 33 (22.2%) were excited, 82 (55.0%) were inhibited, and 34 (22.8%) showed biphasic responses or failed to respond; (2) substitution of low Ca(2+) high Mg(2+) artificial cerebrospinal fluid (aCSF) for normal aCSF abolished the 5 HT-induced inhibitory effect but failed to affect the 5-HT-induced excitatory effect; (3) cyproheptadine, a non-selective 5-HT receptor antagonist, could block either the 5-HT-induced excitatory or inhibitory effects in all neurons tested; and (4) bicuculline, a GABA(A)-receptor antagonist, blocked the 5-HT-induced inhibitory effect. These results imply (1) 5-HT excites arcuate neurons through a mechanism that is insensitive to the decreased extracellular Ca(2+), suggesting a direct postsynaptic action of 5-HT on the 5-HT-receptors located in the membrane of the neurons recorded; and (2) 5-HT might elicit the inhibitory effect through a Ca(2+)-sensitive release of GABA from intercalated GABAergic local neurons that are excited first by 5-HT. PMID- 12075464 TI - [Effect of extracellular chloride concentration on deactivation kinetics of rat ClC-1 chloride channel]. AB - The gating mechanism of ClC-1 chloride channel was studied in this paper by heteroexpression of rat wild type ClC-1 gene in Xenopus oocytes and by two electrode voltage clamping technique. The deactivation gating kinetic parameters were obtained by applying two exponential fitting of the deactivating currents at various extracellular chloride concentrations. It was found that decrease in extracellular chloride concentration increased the fractional amplitude of fast deactivating component, and depressed the fractional amplitude of slow deactivating component accompanied by a decrease in fast and slow deactivating time constants. These results demonstrate that the deactivation kinetic parameters of ClC-1 are largely dependent on the extracellular chloride concentration, which induces changes in channel gating. PMID- 12075465 TI - [Development and progression in rat brain abnormalities related to early stage of epilepsy measured by magnetic resonance image]. AB - The purpose of the present study was to investigate the features of pathophysiological neural networks in rat temporal lobe epileptogenesis. To establish electrogenic epilepsy model, repetitive tetanization (60 Hz, 2 s, 0.4 0.6 mA) was delivered into the right dorsal hippocampus (HPC) of rat brain. Rats were divided into different groups. Experimental animals received tetanic stimulation once a day for 2, 4, 6, 8 or 10 days, respectively. Primary wet dog shakes (WEDS) of the animals were recorded daily during the stimulation to understand the development of behavioral seizures at early stage of epilepsy. The T(2)-weighted (T(2)-WI) spin-echo images were obtained from each experimental rat. The results demonstrated that T(2)-WI hyperintensity of experimental rats was observed in bilateral symmetric dorsal lateral ventricle (LV) areas at stimulating day 2 (n=4), in contralateral medial and ventral LV areas to the side of the electrode at stimulating day 6 (n=5), in contralateral ventral LV areas at stimulating day 8 (n=3), and in ipsilateral ventral LV areas at stimulating day 10 (n=4). Therefore the peak rate of primary WEDS appeared on stimulating day 4 in the experimental rats. Morphological identification demonstrated that the T(2) WI signal abnormalities were related to the enlarged LV and pathological ventricular choroidea plexus hyperplasia. The results suggest that the development of rat brain abnormalities from dorsal LV to ventral LV at early stage of epilepsy can be measured by magnetic resonance image, which implies reorganization of pathophysiologically functional networks before kindling effect appear. PMID- 12075466 TI - [Triggered oscillations in type A dorsal root ganglion neurons induced by veratridine]. AB - Veratridine, a blocker of inactive gate of sodium channel, was used to perfuse L5 dorsal root ganglion (DRG) topically. Afferent activities of type A single fiber from these DRGs were recorded. It was found that after a 10-min bath of veratridine (1.8-3 micromol/L), some of the primary silent DRG neurons were triggered by touch or pressure on the receptive fields or by electrical stimulation of the sciatic nerve to produce high-frequency firing, which was termed triggered oscillation presenting a U-type of interspike intervals (ISI) or other types of oscillations. The longer the intervals between stimulating pulses, the more stimulating pulses were needed to trigger the oscillation. The oscillation, triggered by electric stimuli with different duration or patterns, had no significant difference in their patterns. The duration of the inhibitory period after a triggered oscillation was generally 30-90 s. It was also observed that this kind of triggered oscillation was induced by afferent pulses of the same neurons. These results suggest that triggered oscillation, which may contribute to the fit of triggered pain, can be produced in primary sensory neurons after application of veratridine. PMID- 12075467 TI - [Molecular mechanism of nitric oxide in preventing cardiomyocytes from hypertrophic response induced by angiotensin II]. AB - The aim of this study was to determine the molecular mechanism of nitric oxide (NO) in preventing cardiomyocytes from hypertrophic response induced by angiotensin II (Ang II). Hypertrophic response of neonatal rat cardiomyocytes was assayed by protein synthesis rate and expression of atrial natriuretic peptide (ANP) mRNA. The level of NO was shown by the content of nitrate and nitrite in cardiac myocytes. The protein expression of MKP-1 and the gene expression of eNOS were measured with Western blotting and RT-PCR, respectively. The results are as follows. (1) L-arginine (L-Arg) induced a dose-dependent increase in NO by 16% and 31% at the concentrations of 10 micromol/L and 100 micromol/L, respectively. L-Arg also increased the gene expression of eNOS. However, these effects were inhibited by L-NAME, the inhibitor of NOS. (2) The gene expression and the protein synthesis of ANP induced by Ang II (0.1 micromol/L) were inhibited by L Arg (100 micromol/L). The inhibitory action of L-Arg was abolished after pretreatment with antisense oligoneucleotide against MKP-1. (3) L-Arg (100 micromol/L) increased the protein expression of MKP-1 by 225%, which was inhibited by L-NAME, an NOS inhibitor, and KT-5823, a cGMP-dependent protein kinase (PKG) inhibitor. However, Ang II enhanced the effect induced by L-Arg. The above results show that NO may activate PKG, and thereby promote the protein expression of MKP-1 and inactivate MAPK, resulting in an inhibition of cardiomyocyte hypertrophic response induced by Ang II. PMID- 12075468 TI - Enhancement of Ca(2+) transients and contraction of single ventricular myocytes of rats by 5-(N,N-dimethyl) amiloride. AB - The effects of 5-(N,N-dimethyl)amiloride (DMA) (a blocker of Na(+)/H(+) exchanger or Na(+)/Ca(2+) exchanger) on calcium transient and cell contraction in isolated ventricular myocytes in normal rats and rats with myocardial hypertrophy were examined using ion imaging system with a charge coupled digital camera (CCD camera). Loading myocytes with Fura-2, electrically triggered Ca(2+) transients and cell shortening were measured simultaneously. The results showed that 10 micromol/L DMA increased Ca(2+) transient and cell shortening from 209.60+/-54.96 and 3.07+/-0.97 micrometer to 238.50+/-80.41 and 4.07+/-1.02 micrometer, respectively (P<0.05), which was completely abolished by application of KB-R7943, a specific reverse mode Na(+)/Ca(2+) exchanger blocker. After blocking L-type Ca(2+) channels by nicardipine, DMA also enhanced Ca(2+) transient and cell shortening. In rats with myocardial hypertrophy, DMA showed the common pharmacologic profile as in normal rats but more intense stimulating effects on Ca(2+) transient and cell contraction. The results suggest that DMA increase Ca(2+) transient and cell contraction via stimulating reverse mode Na(+)/ Ca(2+) exchange, and the stimulating effect is more pronounced in rats with myocardial hypertrophy than in normal ones. PMID- 12075469 TI - [Inhibitory effect of alveolar macrophages on the proliferation of pulmonary fibroblasts]. AB - The effect of alveolar macrophages (AM) harvested from Wistar rats by lung lavage on proliferation of human embryo pulmonary fibroblasts in culture was investigated. It was observed that supernatants of AM decreased the uptake of (3)H TdR by the pulmonary fibroblasts. The AM activated with opsonized zymosan (OPZ) showed a stronger inhibitory effect on fibroblast proliferation compared with inactivated AM. Following pretreatment with indomethacin, the inhibitory effect of AM was abolished and reversed to stimulatory effect on pulmonary fibroblast proliferation. The PGE content in AM supernatant was measured with radioimmunoassay. It was observed that the inhibitory effect of AM was highly correlated to prostaglandin (PGE) content in the supernatant of AM. The results suggest that AM has both inhibitory and stimulatory effects on the proliferation of pulmonary fibroblast; the inhibitory effect is primary under normal conditions. This inhibitory action is mainly due to PGE secreted from AM. It is, therefore, suggested that AM plays an important role in suppressing pulmonary fibrosis under normal conditions. PMID- 12075470 TI - Endogenous carbon monoxide attenuates lung injury following ischemia-reperfusion in the hind limbs of rats. AB - To investigate the role of endogenous heme oxygenase (HO)/carbon monoxide (CO) system in the lung injury as assessed by lung histology, polymorphonuclear count, malondialdehyde content and wet-to-dry weight ratio following ischemia reperfusion (I/R) of hind limbs, zinc protoporphyrin (ZnPP), an inhibitor of HO activity, was used, and the lung HO activity and blood carboxyhemoglobin (COHb) level were measured. The results showed that HO activity and COHb level were increased significantly and lung injury occurred after limb I/R. After administration of ZnPP, the lung injury was further aggravated while the HO activity and COHb level were significantly decreased. These findings suggest that upregulation of HO activity followed by subsequent CO production attenuates the lung injury induced by limb I/R in rats. PMID- 12075471 TI - [The effect of nitric oxide on acute lung injury following ischemia/reperfusion of hind limbs in the rat]. AB - On a model of reperfusion after ischemia in the hind limbs (LIR) of rats, we used aminoguanidine (AG) which inhibits nitric oxide synthase (NOS) and L-arginine (L Arg), one of the substrates in the process of nitric oxide synthesis, to observe the changes in NO, NOS, malondialdehyde (MDA), myeloperoxidase (MPO) and wet/dry ratio (W/D) in both skeletal muscles and the lung as well as the changes in phosphatidyl choline (PC) of lung surfactant. The morphologic changes were observed with microscopy. It was observed that the values of NOS, MPO, MDA of the muscle and lung in LIR group increased significantly and the content of PC decreased obviously compared with those of the normal control. Pulmonary observation revealed that after LIR leucocyte assembling and infiltration took place, which was dominated by polymorphocytes with broadened pulmonary interstitial tissue. In LIR+L-Arg group the above changes were reversed, and in LIR+AG group the injuries became more serious. The results obtained suggest that the activity of NOS and the production of NO following ischemia/reperfusion of hind limbs increased significantly, and that the endogenous NO may play a protective role during the early stage of acute lung injury after LIR. PMID- 12075472 TI - Effects of cholecystokinin octapeptide on rat cardiac function and the receptor mechanism. AB - The aim of this study was to explore the effects of cholecystokinin octapeptide (CCK-8) on cardiac function and the receptor mechanism in anesthetized rats. The mean arterial pressure (MAP), heart rate (HR), the left ventricle systolic pressure (LVP) and the maximal/minimum rate of LVP (+/-LV dp/dt(max)) were measured. The results obtained are as follows. (1) Low dose of CCK-8 (0. 4 microgram/kg i.v.) caused tachycardia and slight increase in MAP, LVP and LV dp/dt(max) (P<0.01), while medium dose (4.0 microgram/kg i.v.) and high dose of CCK-8 (40 microgram/kg i.v.) elicited a bradycardia and marked increase in MAP, LVP and LV dp/dtmax (P<0.01). (2) Proglumide (1.0 mg/kg i.v.), a CCK-receptor (CCK-R) antagonist, significantly inhibited the pressor effects of CCK-8, whilst it reversed the bradycardic responses (P<0.01). (3) Using reverse transcription polymerase chain reaction (RT-PCR), CCK-A receptor (CCK-AR) and CCK-B receptor (CCK-BR) mRNA were expressed in myocardium of rats. The above results indicate that CCK-8 may enhance cardiac function in a dose-dependent manner and elicit a change in HR, which is likely induced by the activation of CCK-R on myocardium. PMID- 12075474 TI - [The effect of acetylcholine on the proliferation and apoptosis of three kinds of cultured human pituitary adenoma cells]. AB - In order to elucidate the effect of acetylcholine (ACh) on the occurrence and development of human pituitary adenoma, it was firstly observed whether there exists choline acetyl transferase (ChAT) which is necessary for the synthesis of acetylcholine in the cells of human pituitary adenoma, and then MTT method, (3)H TdR incorporation, cell cycle analysis and TUNEL were employed to estimate the influence of ACh on the proliferation, DNA synthesis and apoptosis of three kinds of human pituitary adenoma (human prolactinoma, somatotropinoma and non functional tumor) cells cultured in vitro. The results showed that (1) the positive staining of ChAT was obviously observed in the cells of the three kinds of human pituitary adenoma, however, it was lower than that in normal human pituitary gland; (2) ACh had a similar effect on the proliferation of the three kinds of human pituitary adenoma cells. ACh at 0.1-10 micromol/L decreased the (3)H TdR incorporation and the MTT A value in a dose-dependent manner. At the same time, ACh decreased the ratio of S or G(2) phase pituitary adenoma cells significantly, but increased the ratio of G(1) phase pituitary tumour cells markedly; (3) the effect of acetylcholine on the proliferation of human pituitary adenoma cells was inhibited by atropine, but not by tubocurarine; (4) ACh had no effect on the apoptosis of human pituitary adenoma cells cultured in vitro. These data suggest that ACh may have a significant modulating effect on the proliferation of pituitary adenoma cells by means of paracrine or autocrine, and the effect is mediated by muscarinic receptor. PMID- 12075473 TI - [IL-1beta stimulates alpha-smooth muscle actin expression through JNK/p38 signal pathway in cultured rat mesangial cells]. AB - To investigate the role of intracellular mitogen activated protein kinase (MAPKs, ERK, JNK and p38) signal pathways in IL-1beta -stimulated alpha-smooth muscle actin (alpha-SMA) expression in rat mesangial cells (rMC), alpha-SMA-promoter gene was transfected into rMC by electro-perforation method and the promoter activity was assayed after IL-1beta (10 ng/ml) stimulation. Protein expression of alpha-SMA was assayed by Western blot. The results were compared between the groups stimulated by IL-1beta with or without PD98059 and SB203580, which are thought to block ERK and p38 pathway, respectively. Dominant-negative-JNKK plasmid was co-transfected in rMC to block JNK pathway. The spatial distribution of alpha-SMA and microfilament-like structure was observed by a confocal laser scanning microscope or an electric microscope. After 6 or 24 h of incubation with IL-1beta, rMC underwent a phenotypic change, which was represented by up regulation of alpha-SMA promoter activity and protein expression. An increase in alpha-SMA and microfilament-like structure was found around the cell nucleus. Block of JNK and/or p38 pathway greatly inhibited IL-1beta -induced alpha-SMA expression, and the block of p38 pathway also suppressed the basal level of alpha SMA expression. In contrast, ERK pathway had no influence on the process. It is, therefore, concluded that IL-1beta -stimulated expression of alpha-SMA is due to its protein synthesis and cytoskeleton re-organization in activated rMC. Intracellular signal regulation of alpha-SMA expression seems to be mediated mainly by JNK/p38 pathways, but ERK appears to have no effect on the process. PMID- 12075475 TI - [Effects of chronic cadmium loading on the testis and endocrine function of reproduction in male rats]. AB - Sixty healthy Sprague-Dawley male rats were used and divided randomly into control group (group C), cadmium loading group with medium dose (group M) and cadmium loading group with high dose (group H). Groups C, M and H were orally dosed daily with 0, 5 and 10 mg/kg of cadmium for over 6 weeks. Effects of cadmium loading on testis and endocrine function of reproduction in male rats were studied. The results showed that the zinc content decreased slightly in testis and plasma, and the cadmium concentration increased significantly in the testis of groups M and H; while the plasma levels of cadmium and zinc had no obvious difference as compared with those of group C; daily sperm production in the testis of group H decreased markedly during week 3 of cadmium loading, and was significantly lower in groups M and H as compared to that in group C during week 6; alkaline phosphatase (ALP) in group H and lactate dehydrogenase-X (LDH-X) in groups M and H were markedly lower compared to those of group C; plasma testosterone (T) level in both cadmium loading groups decreased and was low or significantly lower than that in group C; follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels had no apparent difference between the three groups. It is suggested that the gradual accumulation of cadmium in testis tissue induced by chronic cadmium loading results in changes in some enzyme activity, a decrease in sperm production, and defect of endocrine function activity in the testis. PMID- 12075476 TI - [Adenovirus-mediated NT3 gene transfer protects spiral ganglion neurons from degeneration after noise trauma]. AB - Numerous studies have shown that the health of spiral ganglion neurons is highly important for hearing. As a trophic factor of spiral ganglion neurons, neurotrophin 3 (NT3) is a potential candidate for prevention of spiral ganglion neuron degeneration in human. In our experiments, efficient transduction and long term expression of foreign gene of cochlea cells has been found with adenovirus carried lacZ gene (Ad-lacZ). A model of guinea pig deafness was made by intense noise exposure, which destroyed the entire organ of Corti in the middle part of the cochlea. Seven days after noise exposure, the animals were anesthetized and 1 10(8) recombinant adenoviral particles were injected into the scala tympani through the round window membrane. Animals inoculated with neurotrophin 3 adenovirus(Ad-NT3) were designated as the experimental group, animals inoculated with Ad-lacZ vector served as the control group. Four weeks after the inoculation of the virus, NT3 immunoreactivity was observed in the Ad-NT3 inoculated group. HE histochemical staining results showed that in the Ad-lacZ injected group, the neuronal degeneration was severer and the density of spiral ganglion neurons was significantly lower than those in the Ad-NT3 injected group. Our results demonstrate that with adenovirus-mediated overexpression NT3 may be developed into a new treatment to prevent secondary spiral ganglion degeneration following the damage to Corti organ. PMID- 12075477 TI - [Effect of SO2 derivatives on sodium currents in acutely isolated rat hippocampal CA1 neurons]. AB - The effect of SO2 derivatives on Na(+) currents was studied in freshly dissociated hippocampal CA1 neurons of rat using the whole-cell patch-clamp technique. The results indicated that SO2 derivatives caused a dose-dependent and voltage-dependent increase in the voltage-activated Na+ currents. The amplitudes of Na(+) currents were increased by 50.59 19.08% and 82.06 18.51% (n=15)by SO2 derivatives at 10 and 100 micromol/L, respectively. The action was frequency independent. The results also showed that SO2 derivatives did not affect the activation process, but changed the inactivation process significantly. Before and after application of 10 micromol/L SO2 derivatives, the half-inactivation voltage was -69.71+/-4.67 and -53.27+/-4.95 mV (n=10, P<0.01), respectively, but the slope factor was not changed. These results imply that SO2 derivatives have neurotoxic effects and that SO2 pollution is probably related to some diseases of central neuronal system. PMID- 12075479 TI - Underdiagnosis of sleep apnea syndrome in U.S. communities. AB - We hypothesize that clinical recognition rates for obstructive sleep apnea hypoapnea syndrome (OSAHS) are influenced by comorbidity and demographic factors. Data on medical disorders, symptoms of sleep disorders, and cardiovascular risk factors gathered from 15,699 individuals in the Sleep Heart Health Study were compared. Participants were classified into three groups: those with a self reported physician diagnosis of OSAHS, those with self-reported physician diagnosed and -treated OSAHS, and those reporting both frequent snoring and daytime sleepiness (two-symptom group). Among all participants, 4.1% reported two symptoms (range across sites: 1.55 to 7.23%), whereas 1.6% reported a physician diagnosis of OSAHS (range: 0.66 to 2.88%) and 0.6% reported physician diagnosis and treatment (range: 0.11 to 0.88%). Recognized OSAHS groups were similar to the two-symptom group in age, having a sleeping partner, measured blood pressure, total cholesterol, and race. In a logistic model that included age along with characteristics found to vary significantly among the three groups (gender, body mass index [BMI], high-density lipoprotein cholesterol levels, hypertension), only male gender and BMI were increased in those with physician-diagnosed and treated OSAHS. We conclude that disparities (especially in women and in those with lower BMI) exist between current recognition rates for OSAHS and the estimated prevalence by symptom report across the United States. PMID- 12075480 TI - The effect of posture and a mandibular protruding device on pharyngeal dimensions: a cephalometric study. AB - The objectives were to evaluate the impact of body posture on cephalometric measures and the level and the extent to which treatment with a mandibular protruding device (MPD) affects pharyngeal width. The study was composed of 77 patients: 50 were diagnosed with obstructive sleep apnea (OSA) and 27 snored. After each patient underwent a baseline medical (including a somnographic registration), dental and stomatognathic examination, an MPD that would protrude the mandible 75% of the maximal protrusion range was fabricated. In a radiographic examination, four cephalograms were taken: two in the upright position with and without the MPD and two in the supine position with and without the MPD. The cephalometric measures focused on the pharyngeal structures. MPD treatment significantly increased the relative pharyngeal area (at the level of the oropharynx and the hypopharynx) by a mean of +89.6 mm2 (P < 0.01) in the upright posture. The relative pharyngeal area, however, was reduced by more than 50% in the supine position compared with the upright position, independent of MPD treatment. In the supine position, compared with upright, most pharyngeal measures decreased significantly; the hyoid was significantly lower and displaced posteriorly, and the area of the velum increased by a mean of +201.2 mm2 (P < 0.001). We conclude that the MPD significantly increased most pharyngeal measures except the linear distance between the hyoid bone and the third vertebra and decreased the area of the velum. PMID- 12075481 TI - The efficacy of a novel tongue-stabilizing device on polysomnographic variables in sleep-disordered breathing: a pilot study. AB - The polysomnographic efficacy of a novel tongue-stabilizing device (TSD) in the treatment of snoring and sleep-disordered breathing (SDB) was evaluated in this pilot study. Six current users of the TSD with SDB underwent polysomnography with and without the TSD in situ in a randomized crossover design. The TSD significantly lowered the frequency of snores per hour slept (61- to 70-dB range) (no TSD: mean = 41/h slept +/- 52 SD; TSD: 8/h slept +/- 16 SD; P = 0.046) but did not alter snoring in the other decibel ranges (all Ps > 0.1). Trends were found for reductions in the frequency of apneas plus hypopneas (no TSD: 26/h slept +/- 17/h slept; TSD: 15/h slept +/- 13; P = 0.06) and oxygen desaturations of 4% or more (no TSD: 10/h slept +/- 10; TSD: 5/h slept +/- 5; P = 0.09). Significant improvements in microarousal frequency with the TSD were found (no TSD: 34/h slept +/- 16; TSD: 22/h slept +/- 14; P = 0.004). Significant reductions in percentage of Stage 1 sleep with the TSD were also demonstrated (no TSD: 10 +/- 3%; TSD: 8 +/- 2%; P = 0.03). The results of this small pilot study indicate that the TSD may be effective in reducing snoring severity and microarousals, with favorable trends for reducing SDB severity in selected individuals. Additional larger prospective studies are required to identify suitable candidates for TSD use in the treatment of snoring and SDB. PMID- 12075482 TI - The characteristics of five patients with obstructive sleep apnea whose apnea hypopnea index deteriorated after uvulopalatopharyngoplasty. AB - The objective of this retrospective, consecutive, case series design study was to determine the number of unselected patients with obstructive sleep apnea (OSA) who deteriorated after uvulopalatopharyngoplasty (UPPP). Sixteen of 27 patients at the Sleep Clinic at Veterans Affairs Medical Center who underwent UPPP for OSA and who completed both a pre- and postpolysomnogram were studied. After comparing the apnea-hypopnea index (AHI) before and after UPPP, three groups of patients were identified: deteriorators, unchanged, and improvers. In five patients (31%), the AHI increased by more than 10% after UPPP (deteriorators); in four (25%), the AHI showed a change in either direction of less than 10% (unchanged); and in the remaining seven (44%), the AHI decreased by more than 10% after UPPP (improvers). The AHI deteriorated in five of 16 (31%) unselected patients with OSA in our clinic population who underwent UPPP. The mean pre-UPPP AHI was lower in the patients who deteriorated relative to all other patients (P = 0.02). We suggest that patients who undergo UPPP should have a post-UPPP polysomnogram to determine whether they have improved or deteriorated after the procedure and that alternative forms of treatment may be needed in some patients. PMID- 12075484 TI - "Vanishing white matter" and ovarian dysgenesis in an infant with cerebro-oculo facio-skeletal phenotype. AB - We describe a girl with intrauterine growth retardation, congenital cataracts, mild dysmorphic features and joint contractures. MRI performed at 1 week and 6 months, demonstrated postnatal cerebral and cerebellar atrophy and loss of white matter. The patient died at 10 months. Post-mortem examination revealed ovarian dysgenesis. There was normal development of neuronal and axonal structures, but lack of myelination in brainstem and cerebellum. Subcortical and deep white matter of the cerebral hemispheres were largely destroyed (cavitated), with preserved, but unmyelinated axons in other brain areas. These findings are compatible with a severe leukodystrophic process of pre- or perinatal onset. The exact pathogenesis is not known. PMID- 12075483 TI - The prevalence, cost implications, and management of sleep disorders: an overview. AB - The International Classification of Sleep Disorders distinguishes more than 80 different disorders, which can be effectively treated. Problems with falling asleep or daytime sleepiness affect approximately 35 to 40% of the U.S. adult population annually and are a significant cause of morbidity and mortality. However, the prevalence, burden, and management of sleep disorders are often ignored or overlooked by individuals and society in general. This leads to an underappreciation and undertreatment of sleep disorders, making this group of illnesses a serious health concern. Sleep medicine is a young discipline, and as such the full implications of treating sleep disorders and the extent of sleep related problems are not well delineated. As a result of high prevalence, severe complications, and concomitant illnesses in untreated cases, the cost implications are immense. The costs can be direct, indirect, related, and intangible. However, relatively little has been published on the economic implications of sleep disorders. Economic analysis can help evaluate available resources to set priorities and maximize management strategies for cost control without sacrificing safety, efficacy, or effectiveness. There has been considerable evidence of the cost-effectiveness of treating patients with obstructive sleep apnea, especially considering its high prevalence, morbidity, mortality, and concomitant health care consumption. We review the economic balance sheet of sleep disorders and conclude that sleep medicine education (among general population and health care professionals) and the availability of diagnostic and therapeutic facilities to treat sleep disorders will reduce the profound socioeconomic implications of untreated sleep disorders. PMID- 12075485 TI - Influence of mutation type and location on phenotype in 123 patients with Rett syndrome. AB - Rett syndrome (RTT) is a neurodevelopmental disorder that almost exclusively affects girls. It is caused by mutations in the MECP2 gene that encodes the methyl-CpG-binding protein 2 (MeCP2). In this study we correlated mutation type and location with the severity of the phenotype in 123 girls with RTT. The ability to sit, walk, speak, hand function, head growth, occurrence of epilepsy and a combined severity score were assessed in all girls at 5 years of age and then statistically correlated with the results of the molecular genetic tests. We found that patients who carry either missense mutations or deletions located within the hotspot for deletions, an area between the base pairs (bp) 1030 and 1207 of the MECP2 gene, present with a milder phenotype than other patients. We correlated the location of the mutations with the phenotype and found that all mutations that lead to either a complete or partial truncation of the region coding for the nuclear localisation signal (NLS) are associated with a more severe phenotype than other truncating mutations (p = 0.001). We did not find a significant difference between the patients with mutations in the methyl-CpG binding domain (MBD) and those with mutations in the transcriptional repression domain (TRD). We conclude that mutation type and location correlate with the phenotype in Rett syndrome. All mutations that impair the nuclear localisation signal (NLS) are associated with more severe phenotypes. PMID- 12075486 TI - Dysarthria, progressive parkinsonian features and symmetric necrosis of putamen in a family with painful lipomas (Dercum disease variant). AB - We describe painful subcutaneous lipomatosis in four members of a two-generation family. Lipomas appeared in adulthood, were circumscribed, painful on touch and mainly localized to the trunk and proximal parts of the extremities. The disorder was associated with dysarthria, visual pursuit defect and progressive dystonia. MRI showed bilateral increasing cystic lesions in the basal parts of the putamen. No other abnormalities were detected. The lesions corresponded well with the clinical presentation in the patients. Investigation for mitochondrial disease with muscle biopsy and mitochondrial DNA gave normal results. No consistent biochemical changes were found. The disorder in this family was considered to differ from MERRF with lipomatous lesions and multiple symmetric lipomatosis but compatible with a Dercum disease variant. PMID- 12075487 TI - The early markers for later dyskinetic cerebral palsy are different from those for spastic cerebral palsy. AB - Qualitative abnormalities of spontaneous motor activity in newborns and young infants are early predictive markers for later spastic cerebral palsy. Aim of this research was to identify which motor patterns may be specific for later dyskinetic cerebral palsy. In a large, prospectively performed longitudinal study involving four European hospitals we identified twelve cases with the relatively rare condition of dyskinetic cerebral palsy and compared their early motor development with twelve spastic cerebral palsy cases and twelve controls. From birth to the fifth month post-term, all infants were repeatedly videoed and their spontaneous motor patterns, including general movements, were assessed. Until the second month post-term, the infants that later became dyskinetic displayed a poor repertoire of general movements, "arm movements in circles" and finger spreading. Abnormal arm and finger movements remained until at least five months and were then concurrent with a lack of arm and leg movements towards the midline. Later dyskinetic infants share with later spastic infants the absence of fidgety movements, a spontaneous movement pattern that is normally present from three to five months. Qualitative assessment of spontaneous motor patterns enabled us to identify infants at high risk for dyskinetic cerebral palsy early in life. Additionally, we were able to discriminate them from those infants at high risk for later spastic cerebral palsy. This is a matter of significant clinical relevance because the two types of cerebral palsy ask for different management and early intervention. PMID- 12075488 TI - Cerebral white matter involvement in children with mitochondrial encephalopathies. AB - In childhood mitochondrial encephalopathies the common MRI features are bilateral symmetric abnormalities in basal nuclei and brainstem. The presence of diffuse white matter abnormality has been described only in a few cases. Among a series of 110 children with mitochondrial encephalopathies, 8 patients with MR imaging consistent with a leukoencephalopathy were retrospectively evaluated. Diagnosis was based on the recognition of the biochemical defect in muscle homogenate. H-MR spectroscopic imaging was performed in six of them. Biochemical analysis demonstrated a defect of respiratory chain complexes in six patients: complex I in two cases, complex II in two, complex IV in one, multiple complexes defect in one. Pyruvate dehydrogenase deficiency was demonstrated in two patients. MRI showed severe involvement of the brain white matter without significant basal nuclei or brainstem abnormalities. Two patients developed large cystic areas since onset; in two others progressive vacuolisation of affected white matter was seen later in the course of the disease. One patient with pyruvate dehydrogenase deficiency also presented with a diffuse cortical polymicrogyria. H-MR spectroscopic imaging showed a decrease of N-acetylaspartate, choline and creatine with lactate accumulation in five patients, and was normal in one. These findings suggest that mitochondrial disorders should be included in the differential diagnosis of white matter disorders. PMID- 12075489 TI - Hashimoto encephalopathy - is it underdiagnosed in pediatric patients? AB - Hashimoto encephalopathy (HE) is associated with Hashimoto thyroiditis. Clinically it presents with variable symptoms like seizures, neuropsychiatric changes or focal neurological deficits. Autoimmune phenomena are hypothesized for the pathogenesis. HE has mainly been described in the adult population. We present two 14-year-old patients who presented with recurrent seizures and mental decline. SPECT and PET scans showed distinctly pathological changes. Both patients were diagnosed with HE and improved dramatically on steroids. We feel that HE is a rare but important differential diagnosis of encephalopathy also in the pediatric population. As this disease responds well to steroids, we recommend to obtain basic thyroid function tests as well as thyroid antibodies in all cases of unexplained encephalopathy or unexplained status epilepticus. PMID- 12075490 TI - Chorea Huntington: a rare case with childhood onset. AB - Chorea Huntington (CH) is a dominantly inherited, neurodegenerative disease usually with adult onset. The course of CH is characterized by movement disturbances, psychiatric symptoms and it may lead to dementia. Typically death occurs after 10 to 20 years of CH duration. Invariably, the underlying mutation concerns an expansion of a polymorphic (CAG) n stretch in the huntingtin gene. Statistically, larger expansions lead to earlier onset of the disease. We report on a girl with a huntingtin allele of > 140 (CAG) n repeats. Unspecific neurological symptoms were noted at the age of 4.3 years followed by rapid disease progression with psychomotor deterioration. PMID- 12075491 TI - Polyradiculoneuritis with myelitis: a rare differential diagnosis of Guillain Barre syndrome. AB - AIM: To describe the symptoms, signs, findings and prognosis in children suffering simultaneously from polyradiculoneuritis and myelitis. METHODS: Retrospective review of eight patients aged 2 to 13 years out of 210 patients with polyradiculoneuritis reported from 70 hospitals. Diagnostic criteria for polyradiculoneuritis were: flaccid paresis with loss of tendon reflexes, increased CSF protein and slowing of nerve conduction velocity. Criteria for myelitis were: severe and persistent bladder dysfunction, a sharply defined sensory level and/or evolving spastic paresis, with or without myelitic changes in spinal MRI. RESULTS: In the disease's earliest stage it was difficult to differentiate polyradiculoneuritis with myelitis from classical GBS. However, onset was often unusually rapid compared to GBS. Five patients developed a sensory level and seven suffered from severe bladder dysfunction. Four of the six children studied showed focal myelitic changes in MRI. All seven children with sufficient follow-up remained with residual paresis and significant long-term motor deficits. CONCLUSION: Due to its severe long-term prognosis, polyradiculoneuritis with myelitis must be differentiated from classical GBS. In the disease's early stage, the detection of a sensory level, severe bladder dysfunction and an unusually rapid onset can be helpful. The effect of high-dose corticosteroids is not yet clear. After the acute phase, most children require extended rehabilitation. PMID- 12075492 TI - Cerebral venous thrombosis, intraventricular haemorrhage and white matter lesions in a preterm newborn with factor V (Leiden) mutation. AB - We report a preterm newborn who presented extensive cerebral vein thrombosis on MRI but no abnormal neurological signs. The baby underwent MRI as germinal-matrix intraventricular haemorrhage was revealed by a routine ultrasound brain scan performed on day 16; earlier ultrasound scans (day 2, 7, 12) were all normal. Cerebral vein thrombosis was diagnosed at the first MRI scan together with abnormal restriction in diffusion weighted imaging in the frontal white matter parenchyma. Bilateral microcavitations with a linear pattern of distribution reflecting the distribution of medullary veins developed a week later in the same white matter areas where abnormal diffusion weighted imaging was formerly noted. The baby was later found to be heterozygous for factor V Leiden. PMID- 12075493 TI - Dutch patients with progressive encephalopathy with edema, hypsarrhythmia, and optic atrophy (PEHO) syndrome. AB - PEHO syndrome (progressive encephalopathy, peripheral edema, hypsarrhythmia, and optic atrophy) is a neurodegenerative disorder first characterized in Finnish patients. Subsequent reports have occasionally identified cases of PEHO occurring in some other countries. We describe two Dutch children who represent the first reported cases with PEHO syndrome from western Europe. Both of these children showed typical characteristics of the syndrome, including generalized hypotonia and edema of their faces and extremities, profound psychomotor retardation, progressive cerebellar atrophy, and severe epilepsy which initially started as infantile spasms. Our experience has shown that distinguishing cases with true PEHO from those with other, clinically similar disorders requires a firm demonstration of both the presence and the progress of typical neuroradiological findings. Our present cases suggest that 1) PEHO syndrome is not limited to Finnish heritage, and that 2) many more patients with PEHO could be identified with increased knowledge of the syndrome. PMID- 12075494 TI - Altered methylation pattern of the G6 PD promoter in Rett syndrome. AB - Rett syndrome (RTT) is a neurodevelopmental disorder that almost exclusively affects girls. Recently mutations in MECP2, that encodes the methyl CpG binding protein 2 (MeCP2), have been found to cause RTT. MeCP2 has a role in gene silencing. It binds to methylated cytosine in the DNA and recruits histone deacetylases. We studied the methylation pattern of the promoters of two X chromosomal genes, G6 PD and SYBL1, in patients with RTT and in a control group. Both genes undergo X inactivation which correlates with promoter methylation. A 1 : 1 ratio of methylated versus non-methylated alleles was expected. In the control group a median ratio of 47 : 53 of methylated to non-methylated alleles was found at the G6 PD promoter locus. In 22 patients with RTT the median ratio was significantly different, 33 : 67 (p < 0.0001). Analysis of the SYBL1 promoter revealed an almost identical median ratio of methylated versus non-methylated alleles (RTT 47 : 53; control 49 : 51), however, the range was wider in the RTT group (RTT 23 : 77 to 56 : 44; control 43 : 57 to 55 : 45). There was no apparent correlation between G6 PD promoter methylation status and mutations in the MeCP2 gene or the severity of the clinical phenotype in our patient group. The finding of reduced methylation at the G6 PD promoter is an interesting finding and suggests that there could be widespread dysregulation of X chromosomal genes in Rett syndrome. PMID- 12075495 TI - Alpha-sarcoglycan deficiency featuring exercise intolerance and myoglobinuria. AB - An 8-year-old boy was referred for recent onset of easy fatigue. He showed hyperCKemia and mild scapular winging. Muscle biopsy on the quadriceps muscle demonstrated slight fibre size variability. Dystrophin was normally distributed, carnitine palmitoyl transferase and glycolytic enzymes had normal activities. In the following years the patient developed exercise intolerance and myoglobinuria. Immunohistochemistry showed marked reduction of alpha-sarcoglycan, confirmed by Western blotting. Molecular analysis revealed compound heterozygosity with Arg284Cys and Glu137Lys substitutions, corresponding to nucleotide changes C850 T and G409 A in the gene. At present the patient, 20 years old, shows mild proximal weakness with prominent involvement of the paraspinal muscles, dorsal kyphosis and lumbar hyperlordosis. Exercise intolerance and myoglobinuria, already described in Becker muscular dystrophy, should be also considered among the possible presentations of sarcoglycan deficiencies. PMID- 12075496 TI - [Current epidemiology of carcinoma of the esophagus and cardia in Germany]. AB - BACKGROUND AND OBJECTIVE: In the past 20-30 years the incidence of adenocarcinoma (AC) of the esophagus and the cardia has risen in western countries, while that of squamous cell carcinoma (SC) of the esophagus has largely stayed constant. Current data in the epidemiology of these cancers in Germany were analysed. METHODS AND MATERIAL: The data of the epidemiological cancer register of the Saarland (Old State of former West Germany) and the combined registers of the New States of Germany (former East Germany) and Berlin were collected. Because of the excellent registration procedures, data from the Saarland and the New States, Mecklenburg-Pommern, Brandenburg and Saxony were selected for this study. The incidence was calculated and corrected for age in relation to the world population. RESULTS: The incidence of esophageal AC among males in the Saarland rose from 0.22 per 100 000 population to 1.17 between 1986 to 1997, an increase of 432%, while in Brandenburg, Mecklenburg-Vorpommern and Saxomy it rose, between 1977 and 1998, from 0.23 to 0.8 i.e. by 248%. Esophageal SC incidence remained largely constant in both cancer registers since 1986. In 1998 it was 3.31 in the three New States of Germany and 5.05 in the Saarland. In females the incidence was markedly lower. In 1996-1998 fewer than 10% of cases of esophageal cancer were in UICC stage 1 (T 1 N 0 M 0, early carcinoma). Correspondingly the 5-year survival of all patients with esophageal carcinoma in the Saarland was less than 10% in 1971-1995. AC of the cardia also increased. In the three New States of Germany the incidence among males rose from 1.27 in 1977 to 2.18 in 1998, in females from 0.4 to 0.58. During the same period malignant tumors of the entire stomach, including the cardia, decreased in males from 29.5 to 17.1, in females from 14.3 to 9.2. CONCLUSION: In Germany, during the past 15-20 years, the incidence of esophageal AC and presumably also of AC of the cardia has increased. But the incidence of squamous cell carcinoma of the esophagus has remained largely constant. The proportion of early carcinoma and the 5-year survival with esophageal carcinoma remains under 10%. PMID- 12075497 TI - [Information deficits and hesitant acceptance of combined pancreas-kidney transplantation in Germany. A survey at German dialysis centers]. AB - BACKGROUND AND OBJECTIVE: The pancreas kidney transplantation (PKTx) currently is the only treatment for cure for patients with diabetes mellitus Type 1. Despite the availability of organs and the need for therapy the PKTx rate is much lower in Germany than it is in the United States. Is this attributable to an information gap?. MATERIAL AND METHODS: A questionnaire contrasting 16 statements pro and con PKTx was sent to 960 dialysis centres in Germany. RESULTS: 397 out of 960 questionnaires were returned, resulting in an response rate of 41.4 %. Regional representation was established. The central argument against combined PKTx was an assumed lack of organs (89 %). Other con-arguments were assuming a long waiting list (60 %), lack of qualified transplantation centres (40 %) and poor long-time transplant function (38 %). The most important moderator variable of the distribution of correct/incorrect answers was the direct contact to a cooperating transplant centre. Respondents with direct contact (n = 230) were significantly (p < 0.05 to < 0.001) more likely to hold correct answers with respect to 14 out of 16 single PKTx-related statements. The size of the dialysis centre as determined by the number of patients treated did not alter the distribution of correct/incorrect answers. CONCLUSION: The majority of nephrologists treating patients with Type 1 diabetes underestimate the potential of PKTx. More efforts to increase information are necessary. PMID- 12075498 TI - [Henoch-Schoenlein syndrome in an 80-year-old patient]. AB - HISTORY AND CLINICAL FINDINGS: An 80-year old female patient, hospitalised with pneumonia and myocardial infarction, suddenly developed fever, dyspnoea, malaise, arthralgia and crampy abdominal pain. On the following day, she developed a petechial rash and on the 6 (th) day, macroscopic haematuria. On physical examination, the patient presented with a temperature of 40 degrees C, tachypnoea, tachycardia, and on auscultation coarse crepitations at the left lower base. The abdomen was tender upon deep palpation but bowel sounds were normal. Initially the petechial rash was located at the trunk, later progressing towards the extremities. INVESTIGATIONS: Chest X-ray and laboratory tests were consistent with left lower lobe pneumonia. The clinical picture was not consistent with any haematological causes, coagulopathy or any medication the patient was taking. Rheumatological blood tests (involving ANA, ANCA, rheumatoid factors) were basically negative. A skin biopsy revealed microvessel necrosis and IgA deposits, verifying Henoch-Schoenlein Purpura. An immune electrophoresis revealed a selective IgG-deficiency, which may explain the cause for the patient's recurrent pneumonias. COURSE: The patient recovered well under antibiotic treatment and skin lesions healed without sequelae. CONCLUSION: We present an unusual case of Henoch Schoenlein purpura associated with strictly petechial rash in an 80 year old female. PMID- 12075499 TI - [Chordomas--diagnostic steps and therapeutic consequences]. PMID- 12075501 TI - [The use of human tissue in research. Ethical and legal aspects]. PMID- 12075500 TI - [Dietary prevention of carcinomas of the breast and prostate: fundamental and practical aspects of the Nutritional Cancer Prevention (NCP) program]. PMID- 12075502 TI - [Is an Achilles tendon rupture without degeneration possible?]. PMID- 12075505 TI - [Is the life-long administration of marcumar to a young woman with recurring thromboembolic events necessary?]. PMID- 12075508 TI - Equine granulocytic ehrlichiosis: a case report with DNA analysis and species comparison. PMID- 12075506 TI - Mandibuloacral dysplasia is caused by a mutation in LMNA-encoding lamin A/C. AB - Mandibuloacral dysplasia (MAD) is a rare autosomal recessive disorder, characterized by postnatal growth retardation, craniofacial anomalies, skeletal malformations, and mottled cutaneous pigmentation. The LMNA gene encoding two nuclear envelope proteins (lamins A and C [lamin A/C]) maps to chromosome 1q21 and has been associated with five distinct pathologies, including Dunnigan-type familial partial lipodystrophy, a condition that is characterized by subcutaneous fat loss and is invariably associated with insulin resistance and diabetes. Since patients with MAD frequently have partial lipodystrophy and insulin resistance, we hypothesized that the disease may be caused by mutations in the LMNA gene. We analyzed five consanguineous Italian families and demonstrated linkage of MAD to chromosome 1q21, by use of homozygosity mapping. We then sequenced the LMNA gene and identified a homozygous missense mutation (R527H) that was shared by all affected patients. Patient skin fibroblasts showed nuclei that presented abnormal lamin A/C distribution and a dysmorphic envelope, thus demonstrating the pathogenic effect of the R527H LMNA mutation. PMID- 12075507 TI - CDH23 mutation and phenotype heterogeneity: a profile of 107 diverse families with Usher syndrome and nonsyndromic deafness. AB - Usher syndrome type I is characterized by congenital hearing loss, retinitis pigmentosa (RP), and variable vestibular areflexia. Usher syndrome type ID, one of seven Usher syndrome type I genetic localizations, have been mapped to a chromosomal interval that overlaps with a nonsyndromic-deafness localization, DFNB12. Mutations in CDH23, a gene that encodes a putative cell-adhesion protein with multiple cadherin-like domains, are responsible for both Usher syndrome and DFNB12 nonsyndromic deafness. Specific CDH23 mutational defects have been identified that differentiate these two phenotypes. Only missense mutations of CDH23 have been observed in families with nonsyndromic deafness, whereas nonsense, frameshift, splice-site, and missense mutations have been identified in families with Usher syndrome. In the present study, a panel of 69 probands with Usher syndrome and 38 probands with recessive nonsyndromic deafness were screened for the presence of mutations in the entire coding region of CDH23, by heteroduplex, single-strand conformation polymorphism, and direct sequence analyses. A total of 36 different CDH23 mutations were detected in 45 families; 33 of these mutations were novel, including 18 missense, 3 nonsense, 5 splicing defects, 5 microdeletions, and 2 insertions. A total of seven mutations were common to more than one family. Numerous exonic and intronic polymorphisms also were detected. Results of ophthalmologic examinations of the patients with nonsyndromic deafness have found asymptomatic RP-like manifestations, indicating that missense mutations may have a subtle effect in the retina. Furthermore, patients with mutations in CDH23 display a wide range of hearing loss and RP phenotypes, differing in severity, age at onset, type, and the presence or absence of vestibular areflexia. PMID- 12075509 TI - Diagnostic and hematologic features of probable essential thrombocythemia in two dogs. AB - The clinical and hematologic features of two cases of probable essential thrombocythemia in the dog are described. Both dogs presented with hepatosplenomegaly, severe nonregenerative anemia, neutrophilia and Thrombocytosis. Mean platelet volume and percentages of large platelets were markedly increased in both dogs. Platelet aggregation studies demonstrated hyperaggregability in one dog; platelets from the other dog aggregated spontaneously, precluding further investigation. Cytologic and histologic examination of bone marrow showed pronounced megakaryocytic hyperplasia, with erythroid hypoplasia and relative myeloid hyperplasia. Megakaryocyte morphology was abnormal, with increased numbers of small mononuclear and binucleate cells. Normal to increased hemosiderin stores suggested that apparent macrocytosis in one dog, rather than being due to iron deficiency, resulted from the hematology analyzer counting large platelets as small red blood cells. Megakaryocytic infiltration of the spleen was evident in both dogs. The hematologic findings in dogs with essential thrombocythemia can mimic those associated with iron deficiency anemia, such that diagnostic investigations should be aimed at ruling out chronic blood loss and other causes of reactive Thrombocytosis. PMID- 12075510 TI - Abdominal mass aspirate from a cat with eosinophilia and basophilia. PMID- 12075511 TI - Sensitivity and specificity of cytologic evaluation in the diagnosis of neoplasia in body fluids from dogs and cats. AB - Sensitivity and specificity were determined for the cytologic detection of malignant tumors in canine and feline body cavity effusions. In a prospective study, 424 body cavity effusions from dogs and cats were collected and evaluated, including 70 pleural and 163 peritoneal effusions from dogs, and 77 pleural and 114 peritoneal effusions from cats. Final diagnoses were confirmed in 339 of the 424 cases by clinical follow-up, necropsy, and in the case of malignant tumors, Histopathology. Malignant tumors were found in 18% of canine and 25% of feline body cavity effusions. Approximately one-half of tumors in both dogs and cats were carcinomas. Discrete cell tumors accounted for 56% of feline neoplastic effusions. The sensitivity of cytologic evaluation for the detection of malignant tumors in body cavity effusions was 64% for dogs and 61% for cats. Specificity was 99% for canine and 100% for feline effusions. Sensitivity and specificity were comparable to those obtained with cytologic evaluation of human samples. PMID- 12075512 TI - Deep tissue mass aspirate from a young German shepherd dog. PMID- 12075514 TI - Morphologic classification of anemia. PMID- 12075513 TI - Quality control validation in veterinary laboratories. AB - Quality control (QC) validation is used to determine: 1) whether statistical QC procedures are appropriate for detecting medically important errors; and 2) the equality of performance required by different laboratory tests. QC validation is well documented in the medical literature, but we are unaware of studies addressing its application, problems or unique differences in veterinary laboratories. We applied QC validation to automated hematology and biochemistry analyses in our laboratories, with goals of >/= 90% probability of error detection and 0.80) were achieved between the two test systems for all equine values except MCHC and L+M count, but only for PCV and HB concentration in feline and canine blood samples. PMID- 12075521 TI - Estimation of platelet counts on feline blood smears. AB - Blood samples were obtained from 50 cats admitted for hematologic evaluation at the Royal (Dick) School of Veterinary Studies. Manual platelet counts were done using a hemacytometer, and the average number of platelets per oil immersion field (1,000X magnification) was determined on stained blood smears. A hemacytometer count was not obtained for one sample because of a failure in erythrocyte lysing. In nine samples, obvious platelet clumps in the blood smear prevented accurate determination of the number of platelets per oil immersion field. Hemacytometer counts on these nine samples ranged from 260-587 X 10 (3) platelets/microliter, suggesting that platelet clumps on a blood smear were usually associated with adequate platelet numbers. Simple regression analysis of hemacytometer counts and the average umber of platelets per oil immersion field for the remaining 40 samples yielded correlation coefficients (r) of 0.776 on untransformed data, and 0.892 on log10-transformed data. Each platelet per oil immersion field represented a circulating platelet count of approximately 20 X 10(3)/microliter, similar to conversion factors reported for dogs and human beings. It was concluded that estimation of platelet number on stained blood smears is a simple and quick method that appears to be reliable over a wide range of platelet counts in cats. PMID- 12075522 TI - Fine needle aspirate of a subcutaneous mass on the forelimb of a dog. PMID- 12075523 TI - Spurious platelet effects on erythrocyte indices using the CELL-DYN 3500 automated hematology system. PMID- 12075524 TI - Bone marrow aspirate from a cat with cutaneous lesions. PMID- 12075525 TI - Evaluation of analytical performance assisted by total error criteria of a commercial enzyme immunometric assay for canine serum thyrotropin. AB - The aim of the present study was to evaluate analytical performance using total error criteria of a commercial enzyme immunometric assay for the determination of endogenous canine thyrotropin (TSH). The allowable total error for this assay (22.6%) was estimated using previously reported data on biological variation. Inaccuracy and imprecision of the assay (0% and 5.7% for the low control material; 6.8% and 3.0% for the high control material) were estimated by measuring the same lot of control material for 21 consecutive weeks, during which time the assay was considered stable and in control. Analytical performance was assessed using a MEDx chart, a graphical tool for comparing inaccuracy and imprecision, with an analytical quality requirement stated in the form of allowable total error. The results of the present study showed that the canine TSH assay had good to excellent analytical performance. PMID- 12075526 TI - Myelodysplastic syndrome with sideroblastic differentiation in a dog. AB - A dog with myelodysplastic syndrome had microcytic hypochromic anemia, with siderocytes, poikilocytosis, hypersegmented neutrophils and giant platelets in peripheral blood. In bone marrow, the erythroid series showed immaturity, asynchronous maturation and sideroblasts. Dysgranulopoiesis and dysthrombopoiesis also were present, and hemosiderin was increased. Serum iron concentration was high, and both iron deficiency and lead toxicity were excluded as the cause of dyscrasia. This case represents a unique variant of myelodysplastic syndrome, best described as sideroblastic myelodysplasia. We propose the terms dyserythropoiesis, sideroblastic dyserythropoiesis, dyserythropoiesis with excess blasts, myelodysplasia and sideroblastic myelodysplasia to describe and categorize myelodysplastic syndromes in dogs. PMID- 12075527 TI - Synovial fluid from a kid with polyarthritis. PMID- 12075528 TI - Comparison between microscopic and automated differential leukocyte counts in the silver fox (Vulpes vulpes) and the blue fox (Alopex lagopus). AB - Differential leukocyte (WBC) counts in blood from clinically healthy silver foxes (n=32) and blue foxes (n=37) obtained from an automated hematology analyzer (Technicon H*1 Hematology System) with canine software were compared with microscopic differential WBC counts (M-diff). There was good agreement between the automated differential cell count (A-diff) and the M-diff for neutrophil and lymphocyte percentages. The correlation was lower for monocyte percentages and variable for eosinophil percentages. There was no significant difference between the A-diff and M-diff in either fox species. The A-diff counts were very precise, and may be a good alternative to the traditional M-diff for screening populations of clinically healthy foxes or for studies on stress and animal welfare. Intercept values, however, indicated a constant bias that must be taken into account before interpreting results based on different methods of analysis PMID- 12075529 TI - Peripheral blood from a young lorikeet. PMID- 12075530 TI - Semiautomated analysis of alkaline phosphatase isoenzymes in serum of normal cynomolgus monkeys (Macaca fascicularis). AB - Alkaline phosphatase (ALP) isoenzyme analysis, using a combination of wheat germ lectin (WGL) precipitation, levamisole inhibition and an automated p nitrophenylphosphate assay was validated for use with serum from monkeys (Macaca fascicularis) and used to determine the activities of liver ALP (LALP), bone ALP (BALP) and intestinal ALP (IALP). Based on serial dilution studies and within-run and between-run coefficients of variation, each assay had excellent linearity and acceptable precision. In addition, liver and intestinal mucosa extracts for tissue specific alkaline phosphatases were used to confirm assay validations. Gender-specific differences for total ALP, LALP, and BALP activities were present in sera from normal monkeys between 2 and 4 years of age. Males had 1.3-fold higher total ALP and LALP activities, and 1.5-fold higher BALP activity compared with females. The majority of ALP activity in normal monkey serum was LALP isoenzyme activity, which ranged from 56.7% to 94.7% of total activity. Serum BALP activity ranged from 5.3% to 42.8%. There was negligible IALP activity in all serum samples. PMID- 12075531 TI - Morphology, cytochemical staining and ultrastructural characteristics of reindeer (Rangifer tarandus) leukocytes. AB - Peripheral blood smears from four adult reindeer (Rangifer tarandus) were examined after staining with Romanowsky's stain and cytochemical stains, including alpha-napthyl butyrate esterase (alpha-NBE), Sudan black B (SBB), chloroacetate esterase (CAE) and alkaline phosphatase (ALP). Romanowsky-stained eosinophils, neutrophils, lymphocytes and monocytes resembled those of cattle, sheep and goats. Basophils had two different staining patterns with Romanowsky's stain. Basophils that we termed "grey basophils" were similar in appearance to grey eosinophils in Greyhound dogs, with medium blue-grey to lavender-grey cytoplasm containing varying numbers of clear vacuoles or granules and variable numbers of small, intensely basophilic, perinuclear granules. The second basophil staining pattern was more typical of ruminant basophils, with uniform, pale to dark basophilic cytoplasmic granules. Basophils stained positive for alpha-NBE, SBB, CAE, and ALP. Eosinophils stained positive for SBB, and were negative for alpha-NBE, CAE, and ALP. Neutrophils were negative for SBB, CAE, and ALP. Monocytes stained positive for alpha-NBE, were rarely positive for CAE and SBB, and were negative for ALP. Transmission electron microscopy revealed matrix within all granulocytes granules, including those of basophils. PMID- 12075532 TI - Biological variation of canine serum thyrotropin (TSH) concentration. AB - The aim of the present study was to estimate the between-dog, within-dog and analytical components of variance for serum thyrotropin (TSH) in healthy dogs, and to use these components of variance to 1) estimate the critical difference for significance between serial results; 2) assess the utility of the conventional population-based reference interval; 3) set a desirable performance standard for analytical imprecision; and 4) estimate the number of samples required for determination of the true mean value for an individual dog. Using the Immulite test system, TSH was measured in serum samples collected weekly for five weeks from eight clinically healthy dogs. Results were subjected to nested analysis of variance. Between-dog variation was 43.6%, within-dog variation was 13.6%, analytical variation was 8.8%, the one-sided critical difference was 37.8%, the index of individuality was 0.4, the maximum allowable analytical imprecision was 6.8%, and the number of samples required to determine the true mean value in a single dog was 40. In practical terms, the present study indicated that the analytical imprecision of canine serum TSH measurement should be < 7%, and that comparing a single serum TSH measurement from an individual dog to the conventional population-based reference range may be too insensitive to detect small but important changes in the serum TSH level of that particular dog. In addition, when treating a hypothyroid dog, serum TSH, measured on a weekly basis, should decrease by at least one-third before any effect of exogenous thyroxine supplementation can be said to have influenced the serum TSH level. PMID- 12075533 TI - Evaluation of an automated, homogeneous enzyme immunoassay for serum thyroxine measurement in dog and cat serum. AB - A homogenous enzyme immunoassay (EIA) for measurement of serum thyroxine (T4) concentration was evaluated for use with canine and feline serum. The EIA method was linear from 0 to 150 nmol T4/L for human serum, 0 to 94 nmol T4/L for feline serum and 10 to 60 nmol T4/L for canine serum. Intra- and interassay precision studies yielded coefficients of variation 30-40 WBC or RBC per hpf). Urine parameters that had no effect on efficacy included collection method (cystocentesis or free catch), pH, specific gravity, crystalluria, bilirubinuria, bacteriuria and casts. These data indicated that the Bard BTA test was sensitive for the detection of the bladder tumor-associated antigen complex in canine TCC. As evaluated, this test may serve as a useful adjunct to diagnosis, especially when cytology or biopsy is questionable or impractical. Furthermore, because of the high sensitivity of the test, it may be a practical screening test to rule out TCC in geriatric patients or patients with clinical signs related to the lower urinary tract, particularly before pyuria and hematuria develop which may interfere with test results. PMID- 12075536 TI - "Normal" or reference values: questions and comments. PMID- 12075537 TI - Association of gastrointestinal hemorrhage with increased blood urea nitrogen and BUN/creatinine ratio in dogs: a literature review and retrospective study. AB - Blood urea nitrogen (BUN) and serum creatine concentrations were recorded from the medical records of 52 dogs with clinical evidence for upper gastrointestinal hemorrhage (UGH) at the Colorado State University Veterinary Teaching Hospital between 1988 and 1995. Eighteen of the dogs had melena, 11 dogs had hematemesis, and 23 dogs had both melena and hematemesis. Compared with age-matched control dogs, dogs with suspected UGH had a significantly higher BUN concentration (31.0 mg/dL vs. 16.5 mg/dL), creatinine concentration (1.2 mg/dL vs. 1.1 mg/dL) and BUN/creatinine ratio (27.6 vs. 14.4). The increase in BUN/creatinine ratio suggested that the increased BUN concentration in dogs with evidence for UGH primarily was associated with increased gastrointestinal absorption of nitrogenous compounds, and hepatic ureagenesis. PMID- 12075538 TI - Myelodysplastic changes in a dog with subsequent acute myeloid leukemia. PMID- 12075539 TI - Hematopathology in dogs experimentally infected with a Swedish granulocytic Ehrlichia species. AB - Seven, adult, female beagles were inoculated with a Swedish granulocytic Ehrlichia organism closely related to Ehrlichia equi and E. phagocytophila. Blood and bone marrow changes were evaluated throughout the acute phase of infection. All dogs developed moderate to severe thrombocytopenia during the parasitemic period. The mean platelet volume and platelet distribution width increased, and large platelets were seen on blood smears when platelet numbers were low. In bone marrow, absolute numbers of megakaryocytes and immature megakaryocytes were increased. These results suggested the thrombocytopenia was caused by increased platelet destruction. The dogs also developed mild, normocytic, normochromic anemia, with simultaneous decreases in serum iron concentration and total iron binding capacity that resembled the anemia of inflammation. In bone marrow, there was a slight increase in immature erythroid cells and no erythroid hypoplasia; iron stores were normal to increased. Myeloid hyperplasia was seen in all infected dogs, despite neutropenia in peripheral blood. Lymphopenia occurred early in the parasitemic period, but lymphocytes responded strongly and numbers increased above baseline levels by the end of parasitemia. Blast-transformed lymphocytes (5% to 20%) were seen in peripheral blood for a few days. Experimentally-induced canine granulocytic ehrlichiosis caused cytopenias of short duration, coincident with the appearance of ehrlichial inclusions in neutrophils. PMID- 12075540 TI - Evaluation of piglet blood utilizing the Technicon H*1. AB - The analytic precision of an automated blood analyzer, the Technicon H*1(R), was evaluated utilizing blood samples collected from 20 piglets at 1 and 14 days of age. The effect of storing the blood samples at 4 degrees C for 24 and 48 hours also was determined. Blood samples were analyzed twice on the first day and once on each of the subsequent tow days. Within-sample coefficient of variation was approximately 1% for hemoglobin concentration, erythrocyte count, hematocrit, mean cell volume, erythrocyte distribution width and hemoglobin distribution width (HDW); and approximately 5% for total leukocyte (WBC), neutrophils and lymphocyte counts. Mean HDW and automated differential WBC counts changed during storage to a degree that could be of clinical importance. Manual determination of differential WBC counts were compared with those obtained from the automated analyzer. Results correlated well for neutrophils (r=0.92 in 1-day-old and r=0.93 in 14-day-old piglets, P<0.001) and lymphocytes (r=0.85 in 1-day-old and r=0.93 in 14-day-old piglets, P<0.001). Other WBC values were too low to compare reasonably. PMID- 12075541 TI - The apo-enzyme content of aminotransferases in healthy and diseased domestic animals. AB - We investigated the apo-enzyme content of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in clinically normal and ill canine, feline, equine and bovine patients. Aminotransferase activity was measured with and without the addition of exogenous pyridoxal-5'-phosphate (P5P). The amount of apo enzyme was expressed as the percentage change in aminotransferase activity with the inclusion of P5P. The results of aminotransferase assays without P5P (holo enzyme activity) were highly correlated to the results obtained with P5P (total enzyme activity) in all four species (Spearman rank correlations > 0.980). The median apo-aminotransferase percentage in clinically normal patients was: 11% ALT and 0% AST in 115 dogs, 7% ALT and -5% AST in 50 cats, 6% AST in 46 horses and 9% AST in 50 cattle. The amount of apo-enzyme did not increase as holo-enzyme activity increased in any of the species. The apo-aminotransferase content was not significantly different in canine, feline and equine patients with diseases affecting the liver, kidney or musculature compared to clinically normal animals. The apo-enzyme content did not differ significantly between specific liver diseases in canine and feline patients. PMID- 12075542 TI - Mammary mass aspirate from a Yorkshire terrier. PMID- 12075543 TI - Diagnosis of disseminated adenocarcinoma by bone marrow aspiration in a dog with leukoerythroblastosis and fever of unknown origin. PMID- 12075544 TI - Artifactually increased serum bicarbonate values in two horses and a calf with severe rhabdomyolysis. AB - Extremely high bicarbonate (HCO3-) and anion gap values were measured in two horses and a calf using the Hitachi 911 automated serum biochemistry analyzer. All three animals had severe muscle disease as evidenced by markedly increased aspartate aminotransferase and creatine kinase activities. Laboratory error was suspected as the source of the increased HCO3- because values calculated from blood gas analysis were normal. It was hypothesized that increased serum lactate dehydrogenase (LDH) activity and pyruvate concentration overwhelmed the oxamate LDH inhibitor in the enzymatic HCO3- assay, resulting in consumption of NADH and falsely elevated spectrophotometric reading. Serum LDH activity was markedly increased in all three patients. In an attempt to reproduce this interference in vitro, LDH and pyruvate were added to normal bovine serum. Bicarbonate concentration was artifactually increased in a linear, dose-response relationship proportional to the amount of LDH activity in the sample; addition of pyruvate augmented this increase. It was concluded that increased serum LDH activity and pyruvate concentration secondary to severe muscle disease can result in artifactual increases in serum HCO3- values obtained by routine enzymatic assay. PMID- 12075545 TI - Hematology of equine fetuses with comparisons to their dams. AB - The Hematologic values of 19 equine fetuses between 202 and 238 days gestation were compared with those of their dams. The red blood cell (RBC) count, hemoglobin concentration, hematocrit, and mean corpuscular hemoglobin concentration were significantly lower in fetal blood, while the mean corpuscular volume, mean corpuscular hemoglobin, and red cell distribution width were significantly higher. Mares had a significantly higher nucleated blood cell count than fetuses, and all nucleated cells were leukocytes (WBC). Most WBC in mare blood were segmented neutrophils and lymphocytes. In contrast, over one-half of the nucleated cells in fetal blood were nucleated RBC, and the majority of WBC in fetal blood were lymphocytes. Mares also had significantly higher plasma protein and fibrinogen concentrations than their fetuses. Mild macrocytosis and mild polychromasia were observed in most fetal blood samples, but not in blood samples from mares. All fetal blood contained reticulocytes, and most samples contained Heinz bodies and Howell-Jolly bodies. The results of this study will contribute to the development of hematologic reference values that may be useful in equine fetal research and, possibly, in the diagnosis of equine fetal disease. PMID- 12075546 TI - Serum fructosamine concentration in nondiabetic and diabetic cats. AB - Differentiating transient hyperglycemia from diabetic hyperglycemia can be difficult in cats since single blood glucose measurements reflect only momentary glucose concentrations, and values may be elevated because of stress-induced hyperglycemia. Glycated protein measurements serve as monitors of longer-term glycemic control in human diabetics. Using an automated nitroblue tetrazolium assay, fructosamine concentration was measured in serum from 24 healthy control cats and 3 groups of hospitalized cats: 32 euglycemic, 19 transiently hyperglycemic, and 12 diabetic cats. Fructosamine concentrations ranged from 2.1 3.8 mmol/L in clinically healthy cats; 1.1 - 3.5 mmol/L in euglycemic cats; 2.0 4.1 mmol/L in transiently hyperglycemic cats; and 3.4 to >6.0 mmol/L in diabetic cats. Values for with-in-run precision at 2 fructosamine concentrations (2.64 mmol/L and 6.13 mmol/L) were 1.5% and 1.3%, respectively. Between-run coefficient of variation was 3.8% at a fructosamine concentration of 1.85 mmol/L. The mean fructosamine concentration for the diabetic group differed significantly (P=0.0001) from the mean concentrations of the other 3 groups. Poorly regulated or newly diagnosed diabetic cats tended to have the highest fructosamine values, whereas well-regulated or over-regulated diabetic cats had values approaching the reference range. As a single test for differentiating nondiabetic cats from diabetic cats, fructosamine was very sensitive (92%) and specific (96%), with a positive predictive value of 85% and a negative predictive value of 98%. Serum fructosamine concentration shows promise as an inexpensive, adjunct diagnostic tool for differentiating transiently hyperglycemic cats from poorly controlled diabetic cats. PMID- 12075548 TI - Lymphadenitis and peritonitis caused by Mycobacterium xenopi in a cat. PMID- 12075547 TI - The use of cytochemistry, immunophenotyping, flow cytometry, and in vitro differentiation to determine the ontogeny of a canine monoblastic leukemia. AB - We evaluated the utility of cytochemistry, immunophenotyping, flow cytometry, and in vitro culture with forced differentiation of leukemic cells as diagnostic aids to identify the malignant cell ontogeny in a dog with leukemia. A tentative diagnosis of monoblastic leukemia was established by microscopic examination of Romanowsky-stained blood smears and bone marrow aspirate smears. This diagnosis also was supported by the light scatter signature that identified the blast cells as large, non-granular monocytic cells using a CellDyn 3500 automated hematology analyzer; as well as by the detection of N-butyrate esterase and the lack of choloroacetate esterase or leukocyte peroxidase by cytochemical staining. Subsequently, leukemic cells were isolated from the dog's peripheral blood and placed into tissue culture or cryopreserved. The leukemic cells grew in suspension cultures and proliferated spontaneously for up to 4 days. By day 7, proliferation was negligible. Upon culture with conditioned supernatant using mitogen-stimulated human T cells as a source of cytokines, an increased proportion of cells entered S phase by day 2 of culture; however, proliferation declined markedly by day 4, at which time the cells had apparently differentiated to adherent, vacuolated macrophages. The cytokine-stimulated leukemic cells were positive for the monocyte/macrophage specific markers alpha-1-antitrypsin, alpha 1-antichymotrypsin, lysozyme, CD14, MHC class II, and calprotectin, an antigen found in differentiated macrophages and granulocytes. Despite the strong tendency of the leukemic cells towards monocytic differentiation, our results suggested that they retained some features of a myelomonocytic precursor. These data show that cytochemistry, immunophenotyping, flow cytometry, and in vitro differentiation of canine leukemia cells are useful tools for confirming the lineage of malignant hematopoietic cells. PMID- 12075549 TI - Intra-abdominal mass aspirate from an aged dog. PMID- 12075550 TI - Endogenous estrogen myelotoxicosis associated with functional cystic ovaries in a dog. PMID- 12075551 TI - Peripheral blood and abdominal fluid from a dog with abdominal distention. PMID- 12075552 TI - Automated analysis of feline platelets in whole blood, including platelet count, mean platelet volume, and activation state. AB - A new whole-blood flow cytometric method has been developed for counting and sizing platelets in samples from cats, a species in which platelet and red blood cell sizes overlap significantly. The method is a modified version of the two angle laser light scattering technology used by Bayer H*System hematology analyzers. The new method provided accurate platelet counts and mean platelet volumes (MPV, fl) for cats. The method also measured mean platelet component concentration (MPC, g/dl), a parameter which was shown to be sensitive to platelet activation state, and which decreased in value as activation progressed. PMID- 12075553 TI - Lung aspirate from a cat with respiratory distress. PMID- 12075554 TI - Semiautomatic and robotic methods for determining serum haptoglobin levels. AB - The assay of Makimura and Suzuki for serum haptoglobin has been adapted to use one tube with both semiautomated and robotic methods. The adaptation permits the determination of haptoglobin in several hundred serum samples per day. Both methods were strongly correlated with the original two-tube method and had within assay variability of 4.4% for the semiautomated and 4.8% for the robotic method. Day-to-day variability averaged 6.0% for the semiautomated and 5.5% for the robotic method. The y-intercepts for both assays were higher than that of the original two-tube method, and thus, reference values are higher than those reported previously. PMID- 12075555 TI - Uterine fluid from a mare. PMID- 12075556 TI - Immunophenotypic comparison of blood and lymph node from dogs with lymphoma. AB - Peripheral blood and lymph node tissue from 12 dogs with lymphoma was immunophenotyped. Additionally, the bone marrow was immunophenotyped in 6 dogs. The lymphomas were characterized as B-cell in 11 dogs and T-cell in 1 dog. Immunophenotypic patterns in the peripheral blood and bone marrow were variable. The trend in dogs with B-cell lymphoma was normal to increased percentage of IgG positive cells, decreased percentage of pan-T-positive cells, decreased percentage of CD4-positive cells, and decreased CD4/CD8 ratio. Simultaneous immunophenotyping of lymph node, blood and bone marrow cannot be recommended routinely without further studies to document its value as an independent prognostic indicator. However, it is potentially useful for tumor staging and monitoring remission, especially in lymphoma patients with a leukemic phase. PMID- 12075557 TI - Alanine aminotransferase apoenzyme in dogs. AB - Unusually low serum activity of alanine aminotransferase (ALT) was detected in a Rottweiler dog with gastric dilatation-volvulus. Activity of ALT in the same sample was found to be much higher (estimated increase of 14,225%) when measured by methods adding the cofactor pyridoxal-5'-phosphate (P5P), indicating that nearly all serum ALT was in the apoenzyme form. An investigation was undertaken to determine the frequency of high serum aminotransferase apoenzyme levels in dogs. Eighty canine serum samples submitted to the Clinical Pathology Laboratory at Cornell University were assayed for ALT and aspartate aminotransferase (AST) in the presence or absence of exogenous P5P. In 79 dogs, inclusion of P5P in the LAT assay resulted in a median decrease in AST activity of -6.3% (range -33.3% to 25.0%) in all 80 dogs. One dog had an increase of 336% in ALT activity after inclusion of P5P in the assay, but lacked a similar increase in AST activity. The reason for the high levels of ALT apoenzyme in the 2 dogs was not determined. PMID- 12075558 TI - Transfusion triggers: a systematic review of the literature. AB - Most clinical practice guidelines recommend restrictive red blood cell (RBC) transfusion practices with the goal of minimizing transmission of blood-borne pathogens. The purpose of this review is to compare clinical outcomes in patients randomized to restrictive versus liberal transfusion thresholds (triggers). We conducted a search of OVID Medline, Current Contents, the Cochrane Library, and bibliographies of published studies. Our search strategies used a combination of key-word terms as text and MeSH headings relating to transfusion triggers. We included trials if the comparison groups were assigned on the basis of a clear transfusion trigger or threshold, and the study was randomized with a concurrent control group. Eligibility of studies was assessed by 2 independent raters, with disagreements resolved by consensus. Disagreements not resolved by consensus were referred to a third party for review. Two raters assessed the methodologic quality of the trials modified from the methods of Schultz. The main study outcomes probability of receiving an RBC transfusion, volume of RBCs transfused, hematocrit levels, mortality, and length of hospital stay. Ten trials, which reported outcomes for a total of 1,780 patients, were included. Five studies were in surgical patients, 3 were in the setting of acute blood loss and trauma, and 2 involved intensive care unit patients. Transfusion triggers varied between 7 and 10 g/dL (most often they were 8 or 9 g/dL). Being randomized to a restrictive transfusion trigger group had the following average effects: the probability of receiving an RBC transfusion was reduced by 42% (relative risk, 0.58; 95% confidence interval [CI] 0.47, 0.71), the volume of RBCs was reduced by 0.93 units (95% CI 0.36, 1.5 units), and hematocrit values were 5.6 % lower (95% CI 3.5, 7.7%). Mortality, rates of cardiac events, morbidity, and length of hospital stay were unaffected. The limited published evidence supports the use of restrictive transfusion triggers in patients who are free of serious cardiac disease. However, most of the data on clinical outcomes were generated by a single trial. The effects of conservative transfusion triggers on functional status, morbidity, and mortality, particularly in patients with cardiac disease, need to be tested in further large clinical trials. In countries with inadequate screening of donor blood, the data may constitute a stronger basis for avoiding transfusion with allogeneic RBCs. PMID- 12075559 TI - A review of randomized controlled trials using therapeutic apheresis. AB - Apheresis has developed into a common therapeutic modality. The evidence for the clinical benefit of apheresis, however, is generally from uncontrolled and nonrandomized trials. Although the effectiveness of therapeutic apheresis may be established from uncontrolled trials, well-designed, randomized controlled trials provide the best evidence for a clinical benefit because this type of trial design minimizes bias. We assess the evidence for the use of apheresis, the optimal schedules for apheresis, and the replacement solutions used, based on randomized controlled trials. The databases, MEDLINE and EMBASE, and reference lists from relevant articles were searched. The literature search was restricted to articles published in English and included adult patients only. Two of the authors (NS, CK) independently reviewed the citation list to identify reports for retrieval, and the kappa statistic was used to quantify agreement between the reviewers. Articles were included if they used apheresis as an intervention and had clinical endpoints. The quality of the studies was assessed by using a validated instrument. Five hundred and ninety-two citations were identified, and a total of 85 reports were included in this review. There are only a few well established indications for therapeutic apheresis. Randomized, controlled trials have clearly shown a benefit for patients treated with apheresis who have Guillain-Barre syndrome and patients who have thrombotic thrombocytopenic purpura. A clinical benefit has not been shown for a variety of conditions, including systematic lupus erythematosus, polyarteritis nodosa/Churg-Strauss syndrome, and for the treatment of renal allograft rejection. The effectiveness of apheresis needs to be clarified for several other diseases. As better designed, randomized trials are performed, the role of apheresis for these indications will be further elucidated. PMID- 12075560 TI - Does hybridoma technology still have a place in transfusion medicine? AB - Hybridoma technology has contributed to virtually all areas of biology and medicine and set the scene for important advances in cell biology and immunodiagnostics. Blood transfusion and transfusion medicine have benefited significantly from development in hybridoma technology, and this has resulted in the generation of a wide range of monoclonal antibodies to human blood group antigens. These antibodies have also had an impact on diagnostic techniques such as enzyme-linked immunosorbent assay and flow cytometry as well as on the application of therapeutic agents in the clinical environment. However, rapid advances in the development of knockout and transgenic mice as well as in nucleic acid-based immunization may well lead hybridoma technology into a new era. PMID- 12075563 TI - Proceedings of the NATO Advanced Research Workshop on endocrine disrupters and carcinogenic risk assessment. May 8-12, 2001. Bialystok, Poland. PMID- 12075562 TI - Role of complement in graft rejection after organ transplantation. AB - Activation of the complement system may significantly contribute to the inflammatory reaction after solid organ transplantation. In allotransplantation, the complement system may be activated by ischemia/reperfusion and, possibly, by antibodies directed against the graft. In xenotransplantation from nonprimates to primates, the major activators for complement are preexisting antibodies. Studies in animal models have shown that the use of complement inhibitors may significantly prolong graft survival. This review describes the role of the complement system in organ injury after organ transplantation and the use of complement inhibitors to prevent damage to the graft after allo- or xenotransplantation. PMID- 12075561 TI - Exchange transfusion for malaria and Babesia infection. AB - Malaria accounts for about 2 million deaths per year. Although most cases occur in children in sub-Saharian Africa, fatal infections are seen increasingly in industrialized countries. In 1992, over 900 malaria cases were reported in the United States and a third of these were caused by Plasmodium falciparum. Fatal infections are related to the magnitude of the parasitemia and the immune status of the host. P falciparum poses the greatest threat of death because it invades red cells of all ages, is often drug resistant, and is the only one of the plasmodia species that produces microvascular disease. The risk of death is correlated with the parasite load in immune naive individuals. Babesiosis is generally a subclinical infection in most normal hosts, but it can be life threatening in asplenic patients, older, or immunocompromised individuals. The role of exchange transfusion (ET) in the treatment of these infections is controversial. The Centers for Disease Control recommends that ET be performed in P falciparum infection when parasitemia is equal or greater than 10%. In patients with coma, renal failure, or adult respiratory distress syndrome, ET is recommended regardless of the level of parasitemia even if less than 10%. ET has been advocated to reduce the level of parasitized red blood cells (RBCs), to remove cytokines, and to improve the rheologic properties of the blood. Dramatic improvement has been reported, but there are conflicting reports that question the need for exchange transfusion. This review examines the pathophysiology of severe infection and its treatment, with an emphasis on the role of exchange transfusion. PMID- 12075564 TI - How much are you really worth? Curriculum vitae. PMID- 12075565 TI - Overseas code. PMID- 12075566 TI - No wannabes. PMID- 12075567 TI - Starting blocks. PMID- 12075568 TI - Twins and virtual twins: bases of relative body weight revisited. AB - OBJECTIVE: Application of a new investigative strategy to assess genetic and environmental influences on relative body weight. DESIGN: Covariance structure analysis of body mass index (BMI) using genetically informative samples. SUBJECTS: One-hundred and fourteen monozygotic (MZ) twin pairs (age 5.04-22.93 y), 81 dizygotic (DZ) twin pairs (age 6.05-16.52 y), and 98 virtual twin (VT) pairs, ie same-age unrelated siblings (3.68-54.75 y). MEASUREMENTS: Height, weight, BMI variously obtained by direct measurement and self-report. RESULTS: In contrast with most previous studies, significant common environmental influence on BMI was observed. CONCLUSION: Much past research may have underestimated common environmental effects on BMI because the designs lacked the power or ability to detect them. PMID- 12075569 TI - Calpain 3 gene expression in skeletal muscle is associated with body fat content and measures of insulin resistance. AB - OBJECTIVE: To investigate whether skeletal muscle gene expression of calpain 3 is related to obesity and insulin resistance. DESIGN: Cross-sectional studies in 27 non-diabetic human subjects and in Psammomys obesus, a polygenic animal model of obesity and type 2 diabetes. MEASUREMENTS: Expression of CAPN3 in skeletal muscle was measured using Taqman fluorogenic PCR. In the human subjects, body composition was assessed by DEXA and insulin sensitivity was measured by euglycemic-hyperinsulinemic clamp. In Psammomys obesus, body composition was determined by carcass analysis, and substrate oxidation rates, physical activity and energy expenditure were measured by whole-body indirect calorimetry. RESULTS: In human subjects, calpain 3 gene expression was negatively correlated with total (P = 0.022) and central abdominal fat mass (P = 0.034), and with blood glucose concentration in non-obese subjects (P = 0.017). In Psammomys obesus, calpain 3 gene expression was negatively correlated with circulating glucose (P = 0.013) and insulin (P = 0.034), and with body fat mass (P = 0.049). Indirect calorimetry revealed associations between calpain 3 gene expression and carbohydrate oxidation (P = 0.009) and energy expenditure (P = 0.013). CONCLUSION/INTERPRETATION: Lower levels of expression of calpain 3 in skeletal muscle were associated with reduced carbohydrate oxidation and elevated circulating glucose and insulin concentrations, and also with increased body fat and in particular abdominal fat. Therefore, reduced expression of calpain 3 in both humans and Psammomys obesus was associated with phenotypes related to obesity and insulin resistance. PMID- 12075570 TI - The effect of mild cold exposure on UCP3 mRNA expression and UCP3 protein content in humans. AB - OBJECTIVE: In rodents, adaptive thermogenesis in response to cold exposure and high-fat feeding is accomplished by the activation of the brown adipose tissue specific mitochondrial uncoupling protein, UCP1. The recently discovered human uncoupling protein 3 is a possible candidate for adaptive thermogenesis in humans. In the present study we examined the effect of mild cold exposure on the mRNA and protein expression of UCP3. SUBJECTS: Ten healthy male volunteers (age 24.4 +/- 1.6 y; height 1.83 +/- 0.02 m; weight 77.3 +/- 3.0 kg; percentage body fat 19 +/- 2). DESIGN: Subjects stayed twice in the respiration chamber for 60 h (20.00-8.00 h); once at 22 degrees C (72 degrees F), and once at 16 degrees C (61 degrees F). After leaving the respiration chamber, muscle biopsies were taken and RT-competitive-PCR and Western blotting was used to measure UCP3 mRNA and protein expression respectively. RESULTS: Twenty-four-hour energy expenditure was significantly increased at 16 degrees C compared to 22 degrees C (P<0.05). At 16 degrees C, UCP3T (4.6 +/- 1.0 vs 7.7 +/- 1.5 amol/microg RNA, P=0.07), UCP3L (2.0 +/- 0.5 vs 3.5 +/- 0.9 amol/microg RNA, P=0.1) and UCP3S (2.6 +/- 0.6 vs 4.2 +/- 0.7 amol/microg RNA, P=0.07) mRNA expression tended to be lower compared with at 22 degrees C, whereas UCP3 protein content was, on average, not different. However, the individual differences in UCP3 protein content (16-22 degrees C) correlated positively with the differences in 24 h energy expenditure (r=0.86, P<0.05). CONCLUSION: The present study suggests that UCP3 protein content is related to energy metabolism in humans and might help in the metabolic adaptation to cold exposure. However, the down-regulation of UCP3 mRNA with mild cold exposure suggests that prolonged cold exposure will lead to lower UCP3 protein content. What the function of such down-regulation of UCP3 could be is presently unknown. PMID- 12075571 TI - Relationship between juvenile obesity, dietary energy and fat intake and physical activity. AB - OBJECTIVE: To determine the relationship of juvenile obesity to dietary fat, particularly saturated fat, and with dietary energy (controlling for activity patterns). DESIGN: Cross-sectional, evaluation of diet and activity patterns of obese and non-obese children and adolescents. SUBJECTS: A total of 181 children, aged 4-16 y. Subjects were divided into two groups: obese (body mass index, BMI, > 95th percentile for age and sex), 40 males and 51 females; and non-obese (BMI < 75th percentile for age and sex), 35 males and 55 females. MEASUREMENTS: Dietary intake was analyzed with a dietary history interview; activity patterns were analyzed with an activity interview and body fat was measured with bioelectrical impedance analysis. RESULTS: The obese subjects consumed significantly more total calories, total fat in grams and saturated fatty acids (SFA) in grams than did the non-obese subjects. Based on step-wise multiple regression, the total energy consumed, not total fat or SFA, had the strongest relationship to the subject's percentage body fat, controlling for activity levels. CONCLUSION: We suggest that, although obese children and adolescents consume more dietary energy and fat than non-obese children and adolescents, there is a stronger relationship between total energy consumed and juvenile adiposity than with dietary fat or type of dietary fat consumed. PMID- 12075572 TI - Association between past infection with Chlamydia pneumoniae and body mass index, low-density lipoprotein particle size and fasting insulin. AB - BACKGROUND: Chlamydia pneumoniae infection has been linked to the development of coronary heart disease (CHD), but its relationship to CHD risk factors is less clear. OBJECTIVE: To determine the relationship between past infection with C. pneumoniae and risk factors for CHD, including body weight amongst subjects with and without CHD. METHODS: Antibodies to C. pneumoniae and a range of CHD risk factors were determined in 170 subjects, of whom 43 had recent onset angina. Anthropometric, haemodynamic, lipid and metabolic measurements were obtained and related to antibody status in univariate and multivariate analyses. RESULTS: IgG seropositive (n = 62) did not differ from seronegative subjects in age but were significantly heavier (26.6 +/- 0.4 vs 25.5 +/- 0.3 kg/m2, P = 0.02). The prevalence of seropositivity was similar for subjects with and without CHD and for those with or without hypertension. Subjects with fasting insulin levels greater and those with LDL diameters below the median also had a significantly higher prevalence of seropositivity (45.3 vs 27.3%, P = 0.015 and 45.0 vs 29.4%, P = 0.045 respectively). However in multivariate analysis only body mass index remained significant (P < 0.05). Results were not explained by differences in socioeconomic class. CONCLUSION: Although the study has failed to find a greater prevalence of antibodies to C. pneumoniae amongst subjects with recent onset angina there were associations with a number of cardiovascular risk factors. An increase in body weight appears to underlie these relationships. PMID- 12075573 TI - Relationship between serum magnesium levels and C-reactive protein concentration, in non-diabetic, non-hypertensive obese subjects. AB - OBJECTIVE: To examine the association between serum magnesium levels and C reactive protein (CRP) in non-diabetic, non-hypertensive obese subjects. DESIGN: Cross-sectional study. SUBJECTS: A total of 371 subjects, 101 men and 270 women. Of them 138 lean (37.2%), 133 (35.9%) overweight, and 100 (26.9%) were obese, matched by age. MEASUREMENTS: Fasting and 2 h serum glucose following a 75 g oral glucose load. Fasting serum total cholesterol, HDL- and LDL-cholesterol, triglycerides, C-reactive protein (CRP), albumin; and magnesium levels; urinary protein excretion; body mass index (BMI), waist-to-hip ratio (WHR), and blood pressure. RESULTS: The presence of CRP was documented in four (2.9%) lean, 13 (9.8%) overweight, and 20 (20.0%) obese subjects, and decreased magnesium levels (equal or less than 1.8 mg/dl), in 2 (1.45%) lean, 7 (5.2%) overweight, and 19 (19%) obese subjects. The lowest serum magnesium levels and the highest CRP concentrations were documented in the obese subjects. Twenty-three (82.1%) of the subjects with low serum magnesium (five overweight and 18 obese) showed CRP concentration equal or more than 10 mg/l. There was a graded significant decrease between CRP concentration and serum magnesium levels (r = -0.39, P = 0.002). The odds ratio (CI95%) between magnesium and CRP adjusted by age, sex, BMI and glucose tolerance status for the subjects within the low quartile of magnesium distribution was 2.11 (1.23-3.84). CONCLUSION: The results of this study show that low serum magnesium levels are independently related to elevated CRP concentration, in non-diabetic, non-hypertensive obese subjects. PMID- 12075574 TI - Adiposity, insulin and lipid metabolism in post-menopausal women. AB - OBJECTIVE: To investigate relationships between body fat and its distribution and carbohydrate and lipid tolerance using statistical comparisons in post-menopausal women. DESIGN: Sequential meal, postprandial study (600 min) which included a mixed standard breakfast (30 g fat) and lunch (44 g fat) given at 0 and 270 min, respectively, after an overnight fast. SUBJECTS: Twenty-eight post-menopausal women with a diverse range of body weight (body mass index (BMI), mean 27.2, range 20.5-38.8 kg/m2) and abdominal fat deposition (waist, mean 86.4, range 63.5 124.0 cm). Women with BMI < 18 or > 37 kg/m2, age > 80 y and taking hormone replacement therapy (HRT) were excluded. MEASUREMENTS: Anthropometric measurements were performed to assess total and regional fat deposits. The concentrations of plasma total cholesterol, high density lipoprotein (HDL) cholesterol, triacylglycerol (TAG), glucose, insulin (ins), non-esterified fatty acids (NEFA) and apolipoprotein (apo) B-48 were analysed in plasma collected at baseline (fasted state) and at 13 postprandial time points for a 600 min period. RESULTS: Insulin concentrations in the fasted and fed state were significantly correlated with all measures of adiposity (BMI, waist, waist-hip ratio (W/H), waist-height ratio (W/Ht) and sum of skinfold thickness (SSk)). After controlling for BMI, waist remained significantly and positively associated with fasted insulin (r=0.559) with waist contributing 53% to the variability after multiple regression analysis. After controlling for waist, BMI remained significantly correlated with postprandial (IAUC) insulin (r=0.535) contributing 66% of the variability of this measurement. No association was found between any measures of adiposity and glucose concentrations, although insulin concentration in relation to glucose concentration (glucose-insulin ratio) was significantly negatively correlated with all measures of adiposity. A significant positive correlation was found between fasted TAG and BMI (r=0.416), waist (r=0.393) and Ssk (r=0.457) and postprandial (AUC) TAG with BMI (r=0.385) and Ssk (r=0.406). A significantly higher postprandial apolipoprotein (apo) B-48 response was observed in those women with high BMI (> 27 kg/m2). Fasting levels of NEFA were significantly and positively correlated with all measures of adiposity (except W/H). No association was found between cholesterol containing particles and any measure of adiposity. CONCLUSION: Hyperinsulinaemia associated with increasing body fat and central fat distribution is associated with normal glucose but not TAG or NEFA concentrations in postmenopausal women. PMID- 12075575 TI - Health-related quality of life in WHO class II-III obese men losing weight with very-low-energy diet and behaviour modification: a randomised clinical trial. AB - OBJECTIVE: To study health-related quality of life responses to marked weight loss in WHO Class II-III (body mass index (BMI) > or = 35 kg/m2) obese men. DESIGN: An 8 month randomised clinical trial with a 4 month weight loss programme (10 weeks on a very-low-energy diet (VLED) and 17 behaviour modification visits) in the treatment group and no intervention in the control group. SUBJECTS: Nineteen men (mean age 45.9 y, mean BMI 39.3 kg/m2) in the treatment group and 19 men (47.2 y, 39.4 kg/m2) in the control group. MEASUREMENTS: Weight and questionnaires measuring health-related quality of life (RAND 36-Item Health Survey 1.0 and obesity-related psychosocial problems scale). RESULTS: In the treatment group, the mean (s.d.) weight loss was 17.0 (7.4)% at the end of the 4 month therapy. At the end of follow-up, nearly 6 months after the end of VLED in the treatment group, the average maintained weight loss was 13.9 (7.8)% of baseline weight. The control group was weight stable throughout the study. During treatment, there was only transient improvement in general health, bodily pain, mental health, emotional role functioning and vitality (all increases in the scores were not statistically significant). Improvements in physical functioning, social functioning and obesity-related psychosocial problems were maintained until the end of follow-up. The treatment group also reported improvement in perceived health in the past year. There was only minor fluctuation in questionnaire scores in the control group. CONCLUSION: The short-term and maintained health-related quality of life effects of weight loss may differ. Marked weight loss in WHO Class II-III obese men leads to improvements in physical functioning, social functioning, obesity-related psychosocial problems, and perceived health; these improvements were maintained at 4 month post intervention follow-up. PMID- 12075576 TI - Obesity is associated with decreasing levels of the circulating soluble leptin receptor in humans. AB - OBJECTIVE: Leptin plays a major role in the regulation of body weight. It circulates in both free and bound form. One of the leptin receptor isoforms exists in a circulating soluble form that can bind leptin. In the present study, we measured the soluble leptin receptor (SLR) levels in lean and obese humans. We investigated the relationship between plasma SLR levels, plasma leptin levels and the degree of obesity. We also examined whether SLR concentrations could be modulated by fat mass loss induced by a 3 month weight-reducing diet. SUBJECTS: A total of 112 obese (age 18-50 y; body mass index (BMI) 30-44 kg/m2; 23 men and 89 women), 38 overweight (age 19-48 y; BMI 25-29 kg/m2; 10 men and 28 women) and 63 lean (age 18-50 y; BMI 17-24 kg/m2; 16 men and 47 women) humans. MEASUREMENTS: A direct double monoclonal sandwich enzyme-linked immunosorbent assay (ELISA) was used for the quantitative measurement of the soluble human leptin receptor. Leptin was measured by radioimmunoassay (RIA). Body composition was assessed by biphotonic absorptiometry DEXA (dual energy X-ray absorptiometry). RESULTS: We observed that the SLR is present in human plasma (range 10-100 ng/ml). SLR levels were lower in obese and overweight than lean subjects (28.7+/-8.8, 40.2+/-14.9, 51.2+/-12.5 ng/ml, respectively) and were inversely correlated to leptin and percentage of body fat (r=-0.74 and r=-0.76; respectively; P<0.0001). The ratio of circulating leptin to SLR was strongly related to the percentage of body fat (r=0.91; P<0.0001). Interestingly a gender difference was observed in SLR levels, which were higher in obese and overweight men than in obese and overweight women. In obese subjects after a 3 month low-calorie diet, SLR levels increased in proportion to the decrease in fat mass. In the gel filtration profile, SLR coeluted exactly with the bound leptin fractions. CONCLUSION: Obesity, in humans is associated with decreasing levels of the circulating soluble leptin receptor (SLR). The relationship of SLR with the degree of adiposity suggests that high SLR levels may enhance leptin action in lean subjects more than in obese subjects. PMID- 12075577 TI - The effect of pegylated recombinant human leptin (PEG-OB) on weight loss and inflammatory status in obese subjects. AB - OBJECTIVE: To investigate whether weekly subcutaneous administration of 60 mg of long-acting pegylated human leptin (PEG-OB) for 8 weeks was able to influence weight loss, metabolic profile and inflammatory status of obese subjects on a mildly hypoenergetic diet (deficit: 3.2 MJ/day). DESIGN: A prospective, randomized, double-blind and placebo-controlled single-center trial. SUBJECTS: Twenty-eight healthy, obese subjects (16 women, 12 men; age 22-65 y; body mass index 27.7-38.7 kg/m2). MEASUREMENTS: Bodyweight, metabolic profile (including lipids), C-reactive protein (CRP) and soluble TNF alpha-receptor (sTNF-R) 55 and 75 levels. RESULTS: At the end of the study no significant differences in the delta or percentage weight loss between the placebo (n = 14) and PEG-OB (n = 14) groups was observed. Also the changes in metabolic profile, CRP, sTNF-R55 and R75 concentrations between the two groups after 8 weeks of treatment did not differ. CONCLUSION: Weekly injection of 60 mg PEG-OB did not lead to additional weight loss after 8 weeks of treatment. Furthermore, PEG-OB administration did not affect the changes in metabolic profile and the inflammatory status of obese subjects. PMID- 12075578 TI - The effect of experimentally induced insulin resistance on the leptin response to hyperinsulinaemia. AB - OBJECTIVE: Insulin is thought to be an important regulator of leptin secretion. However, increasing evidence suggests that insulin-mediated glucose uptake rather than insulin per se regulates circulating leptin concentration. Here, we hypothesised that a reduction of insulin sensitivity, ie insulin resistance, will diminish the stimulatory effect of insulin on leptin secretion as a consequence of decreased insulin-mediated glucose uptake. DESIGN: Changes in serum leptin concentration during 30 hyperinsulinaemic-hypoglycaemic clamps were studied after induction of different levels of insulin resistance in normal-weight men. In 15 subjects insulin sensitivity was reduced by exposing them to a 2.5 h antecedent hypoglycaemia (3.1 mmol/l) induced by a high rate of insulin infusion (15.0 mU/min/kg) on the day before the proper experiment ('ante-hypo' condition). In the other 15 subjects no antecedent hypoglycaemia was induced ('control' condition). The proper experiment on both conditions was a 6 h stepwise hypoglycaemic clamp induced by a constant rate of insulin infusion (1.5 mU/min/kg). SUBJECTS: Experiments were carried out in 30 lean healthy subjects (age, mean +/- s.e.m., 26 +/- 1 y; body mass index, 23.1 +/- 0.6 kg/m2). RESULTS: As expected, glucose demand during the clamp was lower in the ante-hypo condition than in the control condition (gram of glucose infused per kilogram body weight, 1.52 +/- 0.16 vs 2.01 +/- 0.17 g/kg; P < 0.05). During the clamp, leptin levels increased by 25.4 +/- 4.3% in the control condition (P < 0.05), but not in the ante-hypo condition (+4.8 +/- 4.5%; P > 0.25). Thus, serum leptin response to the clamp significantly differed between the two conditions (P < 0.01). Across both conditions, the increase of leptin levels during the clamp was correlated with the amount of glucose infused (r = 0.37; P < 0.05). CONCLUSION: Considering that insulin concentrations were identical during both clamp conditions, the data indicate that experimentally-induced insulin resistance diminishes the stimulatory effect of insulin on leptin secretion. PMID- 12075579 TI - Genetic factors as predictors of weight gain in young adult Dutch men and women. AB - OBJECTIVE: To investigate the association between DNA polymorphisms in several candidate genes for obesity and weight gain. Polymorphisms in these genes may contribute to weight gain through effects on energy intake, energy expenditure or adipogenesis. DESIGN AND METHODS: From two large cohorts in The Netherlands (total 17,500 adult men and women), we compared 286 subjects aged 20-40 y who gained an average of 12.8 kg (range 5.5-47 kg) during a mean follow-up of 6.8 y with 296 subjects who remained relatively constant over the same period with respect to occurrence of several polymorphisms in candidate genes of obesity and some lifestyle factors. Subjects who were dieting, were high alcohol consumers, were pregnant, changed their smoking status recently, or those who suffered from serious illnesses were excluded. Polymorphisms were determined in the LEPR-gene (LEPR Lys109Arg, LEPR Gln223Arg, LEPR Lys656Asn), in the UCP1 gene (A-G mutation at position-3826 5' region), in the UCP2 gene (Ala55Val, 45 bp Ins/Del), in the PPARG2 gene (Pro12Ala) and in the ADRB2 gene (Gly16Arg and Gln27Glu). RESULTS: With the exception of the Gly16Arg polymorphism in the ADRB2 gene in men (P = 0.04) and women (P = 0.05), and the Lys109Arg polymorphism in the LEPR gene in women, no statistically significant differences in the genotype and allele frequencies were observed between weight gainers and non-weight gainers. Weight gainers differed in some aspects of dietary habits and physical activity patterns: weight gainers consumed relatively more savory snacks and were less active during leisure time compared with non-weight gainers. CONCLUSION: Only variations in the ADRB2 gene and LEPR gene, may contribute to susceptibility to weight gain. None of the other studied genetic markers were clearly associated with weight gain. Further research is necessary to establish the role of lifestyle factors, or interactions between genes or between genes and lifestyle factors on weight gain with age. PMID- 12075580 TI - Under- and overweight impact on mortality among middle-aged Japanese men and women: a 10-y follow-up of JPHC study cohort I. AB - OBJECTIVE: Although obesity is a major health problem in Western countries, its impact may differ in another culture. This paper examines the association between body mass index (BMI; kg/m2) and mortality in Japan, where the mortality profile and BMI distribution differ substantially from Western countries. DESIGN: The JPHC Study cohort I, a population-based prospective study in four public health center areas, started in 1990 and was followed-up to the end of 1999. SUBJECTS: A total of 19,500 men and 21,315 women aged 40-59 y who submitted their body weight and height and did not report any serious disease at baseline. MEASUREMENT: Risk of death by category of BMI. RESULTS: During 10 y of follow-up, 943 and 483 deaths were documented in the men and women, respectively. The association between BMI and all-cause mortality was U-shaped: compared with a category of 23.0-24.9, the statistically significant elevations in relative risk were observed in both under- and overweight categories (2.26 in 14.0-18.9, 1.57 in 19.0-20.9, 1.33 in 21.0-22.9 and 1.38 in 27.0-29.9, 1.97 in 30.0-39.9 in men, 1.94 in 14.0-18.9 and 1.91 in 30.0-39.9 in women) after adjustment for possible confounders and weight change. The U-shaped association did not change after excluding deaths occurring in the first 5 y of follow-up or even after excluding subjects who reported a weight loss of 5 kg or more since 20-y-old, or after stratifying subjects by smoking status. CONCLUSION: Both under- and overweight are important determinants of premature death among the Japanese population. PMID- 12075581 TI - Temporal trends in overweight and obesity in Canada, 1981-1996. AB - OBJECTIVE: To assess changes in the prevalence of overweight and obesity among Canadian children and adults between 1981 and 1996 using recent recommendations for the classification of overweight and obesity. DESIGN: Epidemiological study comparing the prevalence of overweight and obesity from the 1981 Canada Fitness Survey (CFS) to the 1996 National Longitudinal Survey of Children and Youth (NLSCY) and the 1996 National Population Health Survey (NPHS). SUBJECTS: Adults 20-64 y of age and children 7-13 y of age from the CFS, NLSCY and NPHS. MEASUREMENTS: BMI was calculated from directly measured or self-reported body mass and height. For adults 20-64 y of age, overweight and obesity were defined as BMI > or = 25 kg/m2 and BMI > or = 30 kg/m2, respectively. Age- and sex specific cut-off points for children that correspond to the adulthood categories were used to define overweight and obesity for children 7-13 y of age. RESULTS: The prevalence of overweight increased from 48 to 57% among men and from 30 to 35% among women, while the prevalence of obesity increased from 9 to 14% in men and from 8 to 12% in women. The corresponding increases were from 11 to 33% in boys and from 13 to 27% in girls for overweight and from 2 to 10% in boys and from 2 to 9% in girls for obesity. CONCLUSION: The results indicate dramatic increases in the prevalence of both overweight and obesity in Canada over the last 15 y, and the problem is particularly pronounced among children. PMID- 12075582 TI - Secular trends in overweight and obesity among Finnish adolescents in 1977-1999. AB - OBJECTIVE: To study the trends in overweight and obesity among Finnish adolescents in 1977-1999. DESIGN: Mailed surveys every other year. SUBJECTS: Nationally representative samples of 12, 14, 16 and 18-y-olds (n = 64,147, response rate 78.9%). METHODS: Overweight and obesity were measured by body mass index (BMI) and relative weight (RW) based on self-reported height and weight. BMI > or = the 85th percentile cut-off point for BMI in each age- and sex specific group in the entire data set was considered as overweight, and BMI > or = 95th percentile cut-off point as obesity. RW > or = 110% and > or = 120%, calculated as the individual's weight divided by the mean weight in each age- and sex-specific height percentile group in the entire data set, were considered as overweight and obesity, respectively. The trends in overweight and obesity are described by the change in the 85th and 95th percentile cut-off points of BMI over time. The prevalence of overweight and obesity is also reported using BMI reference values recommended for international comparisons. Because of the similarity of the BMI and the RW criteria in classifying adolescents as overweight and obese, only results based on BMI are presented. RESULTS: Overweight and obesity increased linearly in all sex and age groups from 1977 to 1999. Depending on the age group, the average increase in the 85th percentile cut off point of the BMI per 10 y was 0.6-1.1 kg/m2 in boys and 0.3-0.7 kg/m2 in girls. The 95th percentile cut-off point of the BMI for boys and girls increased by 1.1-1.6 kg/m2 and by 0.6-1.0 kg/m2 per 10 y, respectively. In boys, the increase in overweight and obesity was largest in the two youngest age groups. In girls, the increase in overweight was largest in the oldest age group, and that of obesity both in the 14 and 18-y-olds. Overweight and obesity increased more in boys than in girls in all age groups except in the 18-y-olds among whom the increase was similar in both sexes. Examination of the entire BMI distribution showed that there was little or no change over time at the lower (5th, 15th) and middle (50th) percentiles, but increasing differences at the upper end of the distribution, the increases in the 95th percentile being even more marked than those in the 85th percentile curves. According to international reference values, the age-standardized prevalence of overweight increased in boys from 7.2 to 16.7%, and in girls from 4.0 to 9.8%, between 1977 and 1999. The prevalence of obesity in boys was 1.1% in 1977 and 2.7% in 1999, and in girls 0.4 and 1.4%, respectively. CONCLUSION: Overweight and obesity increased remarkably among Finnish adolescents from 1977 to 1999. The changes concentrated at the upper end of the BMI distribution, suggesting that factors behind this development have influenced only a part of the adolescent population. PMID- 12075583 TI - Time trends of obesity in pre-school children in China from 1989 to 1997. AB - OBJECTIVE: To examine predictors and time trends of obesity in pre-school children in China. DESIGN: This study was based on data from China Health and Nutrition Survey, a longitudinal survey from 1989 to 1997. SUBJECTS: For cross sectional analysis, we included 944, 1058, 903 and 483 children aged 2-6 y in 1989, 1991, 1993 and 1997, respectively. For longitudinal analysis, we included 944 children and 3146 measurements during four time periods. MEASURES: Overweight and obesity according to age- and sex-specific BMI cut-off points proposed by International Obesity Task Force. RESULTS: The overall prevalence of obesity increased from 4.2% in 1989 to 6.4% in 1997 among children aged 2-6 y. The increase largely occurred in urban areas, where the prevalence of obesity increased from 1.5% in 1989 to 12.6% in 1997 and prevalence of overweight increased from 14.6 to 28.9% at the same period. Longitudinal analysis shows BMI increased by 0.2 kg/m2 per year in urban areas and 0.1 kg/m2 per year in rural areas. In multivariate analysis, overweight in early childhood (2-6 y), parental overweight, high income and urban areas independently predicted overweight at age 10-14 y. CONCLUSION: A substantial increase in overweight and obesity among children aged 2-6 y was observed in urban areas in China from 1989 to 1997. Overweight in early childhood significantly predicted overweight during adolescence. Urgent public health strategies are needed to prevent childhood obesity in China. PMID- 12075584 TI - Which aspects of socioeconomic status are related to obesity among men and women? AB - OBJECTIVE: This study aimed to investigate the relationships between body weight and fat distribution, and four empirically derived domains of socioeconomic status: employment, housing, migration status and family unit. DESIGN: A population-based study was used. PARTICIPANTS: A total of 8667 randomly-selected adults (4167 men; 4500 women) who participated in the 1995 Australian National Health and Nutrition Surveys provided data on a range of health factors including objective height, weight and body fat distribution, and a range of sociodemographic indicators. RESULTS: Results demonstrated associations for women, after controlling for age, between the employment domain, and body mass index and waist-to-hip ratio. Low status employed women were 1.4 times as likely to be overweight as high status employed women. There were less consistent relationships observed among these factors for men. Relationships between family unit and indicators of body weight and body fat distribution were observed for both men and women, with those who were married, particularly men (OR = 1.6, 95% CI 1.4-2.0), at higher risk of overweight. The migration and housing socioeconomic status domains were not consistently associated with body mass index or waist-to-hip ratio. CONCLUSIONS: These findings indicate that different components of socioeconomic status may be important in predicting obesity, and thus should be examined separately. Future research would benefit from investigating the underlying mechanisms governing the relationships between socioeconomic status domains further, particularly those related to employment and family unit and obesity. PMID- 12075585 TI - Social class differences in overweight of prepubertal children in northwest Germany. AB - OBJECTIVES: To assess social class differences in overweight and health-related behaviours in 5-7-y-old German children. DESIGN: Cross-sectional study. SETTING: Twenty-nine primary schools in Kiel (inhabitants: 248000), northwest Germany. SUBJECTS: A total of 1350 German 5-7-y-old children and their parents. MAIN OUTCOME MEASURES: Body mass index (BMI), fat mass and health-related behaviours of the children. Self-reported height and weight of their parents, parental school education as a measure of social class. RESULTS: The prevalence of overweight (> or = 90th percentile of reference) was 18.5%. There was an inverse social gradient (P < 0.01): the highest fat mass was observed in children from low social class. The odds ratios for overweight reached 3.1 (CI 1.7-5.4) in boys and 2.3 (CI 1.2-4.3) in girls, respectively (low vs high social class). Overweight parents (BMI > or = 25 kg/m2) were more likely to have overweight children. Parental overweight enhanced the inverse social gradient. The prevalence of overweight was 37.5% (low social class) vs 22.9% (high social class) in children from overweight parents, respectively. There was an inverse social gradient in unhealthy behaviours. Parental BMI and physical inactivity were independent risk factors of overweight in children. CONCLUSIONS: In 5 to 7-y old children overweight and health-related behaviours are inversely related to social class. Parental overweight enhanced the risk of childhood overweight. The familial effect on body weight is most pronounced in children with low social class. Preventive measures should specifically tackle 'overweight families' from low social class. PMID- 12075586 TI - Metabolic syndrome and aging in Wistar Ottawa Karlsburg W rats. AB - BACKGROUND AND OBJECTIVE: Comparative studies have shown that Wistar Ottawa Karlsburg W (RT1u) rats (WOKW) develop a nearly complete metabolic syndrome with obesity, moderate hypertension, dyslipidemia, hyperinsulinemia, and impaired glucose tolerance up to an age of 28 weeks. Because metabolic data thereafter are missing, WOKW and disease-resistant DA rats were studied for 12 months beginning at an age of 5 months. METHODS: Eighteen male inbred WOKW and DA rats were studied monthly from the 5th to the 17th month of life for traits of the metabolic syndrome such as body weight, body mass index (BMI), serum triglycerides, total cholesterol, leptin, insulin as well as glucose tolerance, 24 h excretion of urine total protein and creatinine including telemetric measurement of blood pressure in six males per each group. RESULTS: Except for serum total cholesterol, the measured values for most traits studied were significantly higher in WOKW than in DA rats at an age of 5 months. At an age of 17 months all traits were significantly elevated in WOKW compared with DA rats. WOKW rats were hypertensive, dyslipidemic, obese, glucose intolerant, hyperinsulinemic and proteinuric. CONCLUSION: Considering the phenotype of the WOKW rat described until now and the fact that the metabolic syndrome in this rat is polygenetically determined, the WOKW rat is the most suitable animal model to study the pathophysiology of the facets of the syndrome. PMID- 12075587 TI - Modulation by leptin of proliferation and apoptosis in vascular endothelial cells. AB - AIM: Plasma leptin concentrations not only correlate with body fat mass, but also with the degree of hypertensive retinopathy. The present study was designed to further examine, whether leptin's proliferative, proangiogenic activity relates to a yet uncovered anti-apoptotic effect. RESULTS: Leptin (10-50 nmol/l) concentration-dependently reduced apoptosis in HUVECs (human umbilical vein endothelial cells), HAVECs (human adult vein endothelial cells) and HMECs (human microvascular endothelial cells) by 20% (P < or = 0.05). These findings were supported by increased expression of the apoptosis inhibitor bcl-2 (+55%, P < or = 0.05) as well as by differential modulation of the respective cell cycle checkpoint genes/proteins p53 (-20%, P < or = 0.01), p21(WAF-1/Cip1) (-23%, P < or = 0.05) and the Retinoblastoma protein (+123%, P < or = 0.01). CONCLUSION: bcl 2 dependent anti-apoptotic action might contribute to leptin's proangiogenic activity and thereby promote the development of vascular proliferative disease in obesity. PMID- 12075588 TI - Obesity: under-diagnosed and under-treated in hospital outpatient departments. AB - OBJECTIVE: This study investigated whether medically significant obesity (body mass index > 30 kg/m2) is recognised effectively in hospital outpatient departments and how those identified as obese are subsequently managed. DESIGN: A retrospective analysis of patients' hospital records (rheumatology n = 108, cardiology n = 257, orthopaedic n = 250) established the reported prevalence of obesity and subsequent referral patterns. In addition, BMI was measured prospectively on a separate cohort (rheumatology n = 188, cardiology n = 203, orthopaedic n = 179) to determine the true prevalence. RESULTS: Generally, obesity management appeared minimal and inconsistent. Retrospective analysis revealed an apparently low rate of obesity (4% cardiology, 5% rheumatology and 3% orthopaedics), whilst the true prevalence was found to be 30% for cardiology, 25.1% for orthopaedics and 20.2% for rheumatology. Although this appears to show a large disparity between the apparent and the true prevalence, it is impossible to establish precisely the degree of under-estimation, as the lack of height measurements (14% only) in the retrospective sample affects the reliability of the apparent prevalence. Further comparison with the general population showed obesity to be particularly common in men attending cardiology clinics. CONCLUSION: An outpatient clinic consultation could be a useful starting point for integrating obesity and disease management, by helping to identify obesity, initiate appropriate referrals and assist in obesity education. PMID- 12075590 TI - Night eating and nosology. PMID- 12075589 TI - IGF2 genotype and obesity in men and women across the adult age span. AB - We studied a previously reported association between the IGF2 gene's ApaI polymorphism and obesity in 500 healthy men and women (19-90 y). We hypothesized that individuals homozygous for the IGF2 A allele (A/A) would exhibit lower body mass, BMI and DEXA-measured fat mass compared to G/G homozygotes. Subjects were categorized as exhibiting the G/G (n = 241), G/A (n = 197) or A/A (n = 62) genotype. Contrary to our hypothesis, no difference was observed in body mass, body mass index (BMI) or fat mass between the G/G and A/A genotype groups in the entire cohort. Surprisingly, Caucasian A/A individuals (n = 427) exhibited significantly higher fat mass compared to Caucasian G/G individuals (P < 0.05). In summary, individuals homozygous for the IGF2 G allele do not exhibit higher body mass, BMI or fat mass compared to A/A individuals; however, Caucasians with the A/A genotype exhibit higher fat mass than G/G individuals. PMID- 12075591 TI - Getting your feet wet in the labor pool. PMID- 12075592 TI - How to seek out a mentor. PMID- 12075593 TI - Long odds for the uninsured. PMID- 12075594 TI - Esophageal cancer incidence in the world. PMID- 12075595 TI - In the eye of the beholder. PMID- 12075596 TI - Should beta blockers be abandoned as initial monotherapy in chronic open angle glaucoma? View 1. PMID- 12075597 TI - Should beta blockers be abandoned as initial monotherapy in chronic open angle glaucoma? View 2. PMID- 12075598 TI - Exploring similarity between peer educators and their contacts and AIDS protective behaviours in reproductive health programmes for adolescents and young adults in Ghana. AB - This analysis explores the similarity between peer educators and their contacts. To examine interpersonal communication in the context of peer education, this study tested a new approach using multiple semi-structured interviews and network analysis to collect data from 106 peer educators and 526 of their contacts. These evaluation activities were conducted at three sites in Ghana during April 1998, in peri-urban and rural locations, and in in-school and out-of-school targeted settings. It was found that in their peer counselling and peer promotion activities peer educators tend to reach people who are like themselves (53% within 2 years of age, 59% same sex, 70% same ethnicity, and 65% same school status) however, this trend is not uniform among all youth and varies by demographic characteristics and their cultural environment. By examining the social networks of peer educators, it is possible to gain a better understanding of the process of peer education counselling in the context in which it occurs. The study also shows that controlling for other factors, contacts of peer educators who are highly similar regarding age, sex, ethnicity, and school status, are 1.74 times more likely (95% CI: 1.18, 2.56) to have done something to protect themselves from AIDS in the past three months. The results have relevance for programme managers and planners, researchers, and international agencies serving youth. PMID- 12075599 TI - Ipsilateral femoral vein compression: a contraindication to thrombin injection of femoral pseudoaneurysms. AB - Development of a femoral artery pseudoaneurysm occurs in 0.6% to 3.2% of interventional procedures. Nonsurgical treatment has consisted of ultrasound scan directed compression and, more recently, direct thrombin injection into the pseudoaneurysm cavity to achieve thrombosis. Reported complications after thrombin injection are rare. We report two cases of femoral venous compression associated with pseudoaneurysm injection and review the literature. A 76-year-old man and an 86-year-old man both underwent thrombin injection of pseudoaneurysms compressing the ipsilateral common femoral vein. Both patients were diagnosed with deep venous thrombosis and subsequently needed surgical exploration for repair of the pseudoaneurysm and release of the venous compression. At exploration, both were found to have significant inflammation surrounding the femoral vessels, which made vessel exposure challenging. Because of the venous outflow obstruction involved in femoral pseudoaneurysms with secondary venous compression and the surgical difficulty caused by surrounding inflammation, avoidance of thrombin injection in favor of early surgical intervention is suggested. PMID- 12075600 TI - Primary aortoappendiceal fistula: case report and review of the literature. AB - We report a case of primary aortoappendiceal fistula in a patient with chronic, relapsing abdominal symptoms and acute lower gastrointestinal hemorrhage. The diagnosis was evident with abdominal computed tomographic scan results. The patient's condition was successfully managed with appendectomy, abdominal aortic aneurysm resection, and extra-anatomic bypass grafting. Review of the literature revealed this to be the first report of a true aortoappendiceal fistula. PMID- 12075602 TI - [Thyroid cancer: epidemiology and predisposing mechanisms due to ionizing radiation]. PMID- 12075601 TI - [Is pamidronate or clodronate better?]. PMID- 12075604 TI - Strabology report of the 28th annual meeting of the American Association for Pediatric Ophthalmology and Strabismus. PMID- 12075603 TI - Effects of energy expenditure and Ucp1 on photoperiod-induced weight gain in collard lemmings. AB - OBJECTIVE: To determine the role of total energy expenditure (TEE) and its components in the ability of collared lemmings to increase weight in response to a decrease in photoperiod. RESEARCH METHODS AND PROCEDURES: Energy expenditure was measured by 24-hour indirect calorimetry concurrent with food-intake studies. TEE and resting and nonresting energy expenditure (REE and NREE, respectively) were adjusted for body weight by analysis of covariance (ANCOVA). Uncoupling protein 1 (Ucp1) mRNA levels from interscapular brown adipose tissue were determined by Northern blot. RESULTS: TEE and REE of lemmings exposed to a short photoperiod for 10 days were significantly lower than that of lemmings exposed to a long photoperiod (p < 0.05), whereas NREE was not significantly different (p = 0.44). Ucp1 mRNA levels in interscapular brown adipose tissue were 50% lower in short- vs. long-photoperiod lemmings (p < 0.01). Ucp1 mRNA levels were positively related to REE (r2 = 0.79, p < 0.01). After adjustment of REE for differences in Ucp1 mRNA levels, there was no longer a significant difference attributable to photoperiod treatment (p = 0.54). DISCUSSION: The results of this study indicate that the increase in body mass that occurs when collared lemmings are exposed to a short photoperiod may be primarily fueled by a decrease in REE and is correlated with a decrease in Ucp1 mRNA levels. PMID- 12075605 TI - Quality nursing care celebrates diversity. PMID- 12075606 TI - Nursing as a context for alternative/complementary modalities. PMID- 12075607 TI - Nursing as a context for alternative/complementary modalities. PMID- 12075608 TI - Domestic violence: the challenge for nursing. PMID- 12075609 TI - Telehealth: promise or peril? PMID- 12075610 TI - "An historical perspective on genetic care". PMID- 12075611 TI - "Nursing shortage: not a simple problem--no easy answers". PMID- 12075612 TI - State Boards of Nursing and scope of practice of registered nurses performing complementary therapies. PMID- 12075613 TI - Abstracts of the 2nd International Conference on Biology, Chemistry and Therapeutics Applications of Nitric Oxide. Prague, Czech Republic, June 16-20, 2002. PMID- 12075615 TI - Abstracts for the Annual Scientific Meeting of the Thoracic Society of Australia and New Zealand. 24-27 March 2002. Cairns, Australia. PMID- 12075614 TI - Early or delayed surgical treatment of supracondylar humeral fractures in children. PMID- 12075616 TI - Abstracts of the 3rd International Conference of the Asia Pacific Clinical Nutrition Society. 10-13 March 2002, Osaka, Japan. PMID- 12075617 TI - Abstracts from the 13th World Congress of the International Society for the Study of Hypertension in Pregnancy. Toronto, Canada, June 2-5, 2002. PMID- 12075618 TI - Abstracts of the 11th Symposium of the AEP Section Epidemiology and Social Psychiatry. 17-20 April 2002. Aarhus, Denmark. PMID- 12075619 TI - Abstracts of the 25th Annual Meeting of the American Society of Primatologists. Oklahoma City, Oklahoma, USA. June 1-4, 2002. PMID- 12075620 TI - From the Centers for Disease Control and Prevention. Assessment of susceptibility testing practices for Streptococcus pneumoniae--United States, February 2000. PMID- 12075622 TI - [Italian Society of Medical Radiology membership directory. December 2001]. PMID- 12075621 TI - From the Centers for Diseases Control and Prevention. Respiratory illness in workers exposed to metalworking fluid contaminated with nontuberculous mycobacteria--Ohio, 2001. PMID- 12075623 TI - Proceedings of the British Pharmacological Society Meeting. 11-12 April 2002. PMID- 12075624 TI - Adenylyl cyclase and G-proteins in Phytomonas. AB - Phytomonas sp. membranes have an adenylyl cyclase activity which is greater in the presence of Mn2+ than with Mg2+. The Mg2+ and Mn2+ activity ratio varies from one membrane preparation to another, suggesting that the adenylyl cyclase has a variable activation state. A[35S]GTP-gamma-S-binding activity with a Kd of 171 nM was detected in Phytomonas membranes. Incubation of these membranes with activated cholera or pertussis toxin and [adenylate 23P]NAD+ led to incorporation of radioactivity into bands of about 40-44 kDa. Crude membranes were electrophoresed on SDS-polyacrylamide gels and analyzed, by Western blotting, with the 9188 anti-alpha[s] antibody and the AS/7 antibody (anti-alpha[i], anti alpha[i1], and anti-alpha[i2]. These procedures resulted in the identification of polypeptides of approximately 40-44 kDa. Phytomonas adenylyl cyclase could be activated by treatment of membrane preparations with cholera toxin, in the presence of NAD+, while similar treatment with pertussis toxin did not affect this enzyme activity. These studies indicate that in Phytomonas, adenylyl cyclase activity is coupled to an unknown receptor entity through G alpha[s] proteins. PMID- 12075625 TI - gamma-Glutamyl transpeptidase and glutathione biosynthesis in non-tumorigenic and tumorigenic rat liver oval cell lines. AB - Glutathione synthesis and growth properties were studied in the gamma-glutamyl transpeptidase(GGT)-negative, non-tumorigenic rat liver oval cell line OC/CDE22, and in its GGT-positive, tumorigenic counterpart line M22. gamma-Glutamylcysteine synthetase (GGCS) activities were comparable. Growth rates of M22 cells exceeded those of OC/CDE22 cells at non-limiting and limiting exogenous cysteine concentrations. A monoclonal antibody (Ab 5F10) that inhibits the transpeptidatic but not the hydrolytic activity of GGT did not affect the growth rates of OC/CDE22, and decreased those of M22 to the OC/CDE22 level. In GSH-depleted M22, but not in OC/CDE22 cells, the rate and extent of GSH repletion with exogenous cysteine and glutamine exceeded those obtained with exogenous cysteine and glutamate. With Ab 5F10, repletion with cysteine/glutamine was similar to that obtained with cysteine/glutamate. Repletion with exogenous GSH occurred only in M22 cells, and was abolished by the GGT inhibitor acivicin. Repletion with gamma glutamylcysteine (GGC) in OC/CDE22 was resistant to acivicin whereas that in M22 was inhibited by acivicin. Repletion with exogenous GSH or cysteinylglycine (CG) required aminopeptidase activity and was lower than that obtained with cysteine. Unless reduced, CG disulfide did not support GSH repletion. The findings are compatible with the notions that (i) GGT-catalyzed transpeptidation was largely responsible for the growth advantage of M22 cells at limiting cysteine concentration, and for their high GSH content via the formation of GGC from a gamma-glutamyl donor (glutamine) and cyst(e)ine, and (ii) aminopeptidase/dipeptidase activity is rate-limiting in GSH repletion when GSH or CG serve as cysteine sources. PMID- 12075626 TI - How it feels to be a child of donor insemination. PMID- 12075627 TI - Education program doubles use of pain management interventions in nursing homes. PMID- 12075628 TI - Best bets for managing insomnia: drug therapy, behavioral interventions. PMID- 12075629 TI - Updated guidelines call for higher colorectal cancer screening rates. PMID- 12075630 TI - Antibiotics unnecessary for most respiratory infections. PMID- 12075631 TI - Intravenous Induction Agents--propanidid. PMID- 12075633 TI - Most nosocomial pneumonias are not due to nosocomial bacteria in ventilated patients. Evaluation of the accuracy of the 48 h time cut-off using carriage as the gold standard. AB - A prospective observational cohort study was undertaken with two endpoints: (1) to compare the time cut-off of 48h and the carrier state criterion for classifying lower airway infections in adult and paediatric long-term ventilated patients, and (2) to evaluate the potential of optimized time cut-offs for characterizing imported and ICU-acquired lower airway infections. All patients admitted to the general and paediatric intensive care units and expected to require mechanical ventilation for a period > or = 3 days were enrolled. Surveillance cultures of throat and rectum were obtained on admission and thereafter twice weekly to distinguish micro-organisms that were imported into the unit from those acquired during the stay on the unit. A total of 130 adults and 400 children were studied. In the adult population, 70% of lower airway infections were classified as ICU-acquired by the 48 h cut-off and 48% by the criterion of carriage; on the paediatric ICU the percentages were 65% and 20%, respectively. To separate imported from ICU-acquired infections, eight days was optimal in the adult population and 10 days in the paediatric population. Sensitivity, specificity, positive predictive value and negative predictive value for a time cut-off of eight days for adults were 86, 77, 80, 83%, respectively, and using 10 days for children were 87, 62, 90, 56%, respectively. The use of the 48 h cut-off rule classifies patients as having nosocomial pneumonia, when in fact the infections are commonly caused by microorganisms carried in by the patients. In contrast, using the carriage method, the proportion of lung infections due to nosocomial bacteria was relatively small and was a late phenomenon. Although in prolonging the time cut-off the difference between the two types of classification was shorter, time cut-offs were still found to be unreliable for distinguishing imported from unit-acquired lower airway infections. PMID- 12075632 TI - Procalcitonin and proinflammatory cytokine clearance during continuous venovenous haemofiltration in septic patients. AB - Procalcitonin (PCT), interleukin-6 (IL-6), tumour necrosis factor a (TNFalpha), and interleukin-1beta (IL-1beta) are important clinical prognostic markers in ICU septic patients. The goal of the study was to determine whether continuous venovenous haemofiltration (CWH), using an AN69 haemofilte, leads to elimination of PCT, TNFalpha, IL-6 and IL-1beta in 13 septic patients with multi-organ failure. At the start of haemofiltration (0), 6 and 12 hours the mean afferent plasma concentration +/- SD of PCT (10.1 +/- 9.1, 7 +/- 6, 5.9 +/- 5.7 ng/ml), IL 6 (804.6 +/- 847.6, 611.7 +/- 528.4, 575.2 +/- 539.2 pg/ml), and that of TNFalpha (4.5 +/- 2.6, 4 +/- 3.1, 3.8 +/- 2.9 pg/ml) significantly declined during CVVH. The efferent plasma concentrations were significantly lower than the corresponding afferent concentrations. PCT; IL-6 and TNFalpha were detectable in the ultrafiltrate of all patients. IL-1beta was only detectable in the plasma of eight patients and the ultrafiltrate of five patients. The plasma clearance of PCT, IL-6 and TNFalpha significantly decreased after 12 hours as a result of a decline in the adsorptive elimination of the mediators due to progressive membrane saturation. We demonstrated that if PCT, IL-6 and TNFalpha are used as clinical prognostic markers in septic patients who are treated with CWIH using an AN69 membrane, one should be aware that their plasma level could be modified by the therapy. In addition CWH could represent an appropriate tool to remove a broad spectrum of proinflammatory mediators, if such removal is required in septic patients. PMID- 12075634 TI - The effects of three manual hyperinflation techniques on pattern of ventilation in a test lung model. AB - Manual hyperinflation (MHI) is used by physiotherapists as a treatment technique in intubated patients. This study investigated the effect of three different MHI techniques using a Mapleson-C circuit configuration with a CIG Medishield valve on volume delivered (Vt), peak inspiratory (PIFR) and expiratory flow rates (PEFR), and peak airway pressure (PAP) in a test lung model. The protocols differed in the degree of valve closure and inclusion of an inspiratory pause. For protocols 1, 2 and 3 the measures were Vt-1.33 (0.21), 2.74 (0.13), 3.55 (0.12) litres; PAP-14.30 (0.82), 24.00 (0.47), 30.20 (0.92) cmH2O and PIFR-1.13 (0.05), 1.51 (0.15), 1.32 (0.09) l/s respectively. All pair comparisons were statistically significant except for PEFR (l/s), which was significantly lower for protocol 1 [1.62 (0.06)], compared to protocols 2 [2.01 (0.25)] and 3 [2.10 (0.19)] but not between protocols 2 and 3. Circuit and technique choice should be considered in relation to the specific therapeutic aim of treatment. PMID- 12075635 TI - Survival following mechanical ventilation of recipients of bone marrow transplants and peripheral blood stem cell transplants. AB - Survival of bone marrow transplant recipients requiring mechanical ventilation is poor but improving. This study reports a retrospective audit of all haematopoietic stem cell transplant (HSCT) recipients requiring mechanical ventilation at an Australian institution over a period spanning 11 years from 1988 to 1998. Recipients of autologous transplants are significantly less likely to require mechanical ventilation than recipients of allogeneic transplants. Of 50 patients requiring mechanical ventilation, 28% survived to discharge from the intensive care unit, 20% to 30 days post-ventilation, 18% to discharge from hospital and 12% to six months post-ventilation. Risk factors for mortality in the HSCT recipient requiring mechanical ventilation include renal, hepatic and cardiovascular insufficiency and greater severity of illness. Mechanical ventilation of HSCT recipients should not be regarded as futile therapy. PMID- 12075636 TI - Closed-loop control of blood glucose levels in critically ill patients. AB - A closed-loop control system was constructed for automatic intravenous infusion of insulin to control blood sugar levels (BSL) in critically ill patients. We describe the development of the system. A total of nine subjects were recruited to clinically test the control system. In the patients who underwent closed-loop control of BSL, the controller managed to control only one patient's glycaemia without any manual intervention. The average BSL attained during closed-loop control approached the target range of 6-10 mmol/l, and had less deviation than when BSL had been maintained manually. We conclude that closed-loop BSL control using a sliding scale algorithm is feasible. The main deficiency in the current system is unreliability of the subcutaneous glucose sensor when used in this setting. This deficiency mandates high vigilance during use of the system as it is being developed. PMID- 12075637 TI - Prospective independent validation of APACHE III models in an Australian tertiary adult intensive care unit. AB - Evaluation of the performance of the APACHE III (Acute Physiology and Chronic Health Evaluation) ICU (intensive care unit) and hospital mortality models at the Princess Alexandra Hospital, Brisbane is reported. Prospective collection of demographic, diagnostic, physiological, laboratory, admission and discharge data of 5681 consecutive eligible admissions (1 January 1995 to 1 January 2000) was conducted at the Princess Alexandra Hospital, a metropolitan Australian tertiary referral medical/surgical adult ICU ROC (receiver operating characteristic) curve areas for the APACHE III ICU mortality and hospital mortality models demonstrated excellent discrimination. Observed ICU mortality (9.1%) was significantly overestimated by the APACHE III model adjusted for hospital characteristics (10.1%), but did not significantly differ from the prediction of the generic APACHE III model (8.6%). In contrast, observed hospital mortality (14.8%) agreed well with the prediction of the APACHE III model adjusted for hospital characteristics (14.6%), but was significantly underestimated by the unadjusted APACHE III model (13.2%). Calibration curves and goodness-of-fit analysis using Hosmer-Lemeshow statistics, demonstrated that calibration was good with the unadjusted APACHE III ICU mortality model, and the APACHE III hospital mortality model adjusted for hospital characteristics. Post hoc analysis revealed a declining annual SMR (standardized mortality rate) during the study period. This trend was present in each of the non-surgical, emergency and elective surgical diagnostic groups, and the change was temporally related to increased specialist staffing levels. This study demonstrates that the APACHE III model performs well on independent assessment in an Australian hospital. Changes observed in annual SMR using such a validated model support an hypothesis of improved survival outcomes 1995-1999. PMID- 12075639 TI - The use of glyceryl trinitrate ointment with EMLA cream for i.v. cannulation in children undergoing routine surgery. AB - This prospective randomized controlled double-blind trial was performed to investigate whether glyceryl trinitrate (GTN) ointment, applied ten minutes after the removal of a eutectic mixture of local anaesthetic cream, lignocaine/ prilocaine (EMLA), influences site selection for intravenous cannulation in children. Eighty children aged between five and 15 years, having general surgery as day patients, received EMLA cream on both hands for 90 minutes prior to transfer to the theatre suite. Acting as their own controls, the children received GTN on one hand, and placebo on the other, after removal of the EMLA cream and ten minutes prior to the insertion of an intravenous cannula. The application of GTN or placebo to left or right hand was blinded and randomized. At induction, the hand with the visually best quality veins was selected and cannulated. The primary outcome was selection between the EMLA/GTN hand and EMLA/placebo hand. The selection having been made, the number of attempts at cannulation, ease of insertion and pain scores (VAS) for cannulation were also recorded. The choice of hand was positively influenced by the use of GTN, with the GTN hand chosen in 51 of 72 (70%) children who completed the protocol (P = 0.001). The findings of this study suggest that the application of GTN after EMLA removal may be clinically useful in aiding cannulation in children. PMID- 12075638 TI - Effectiveness of intravenous ephedrine infusion during spinal anaesthesia for caesarean section based on maternal hypotension, neonatal acid-base status and lactate levels. AB - Maternal cardiovascular changes and neonatal acid-base status, including lactate levels, were assessed in 30 healthy women undergoing elective caesarean section under spinal anaesthesia. Patients were allocated randomly to receive IV ephedrine infusion (n = 15) (5 mg.min(-1) immediately after the spinal injection or bolus administration of IV ephedrine (n = 15) (10 mg) in case of development of hypotension. Maternal and neonatal blood pressure, heart rate and acid-base status including lactate levels were compared between the groups. Systolic blood pressure in the bolus group was significantly lower when compared to the infusion group. Nausea was observed in one patient (6%) in the infusion group and nausea and vomiting were observed in 10 patients (66%) in the bolus group. Although umbilical arterial pH values were significantly lower in the bolus group, lactate levels were similar In conclusion, ephedrine infusion prevented maternal hypotension, reduced the incidence of nausea and vomiting and led to improved umbilical blood pH during spinal anaesthesia for caesarean section. PMID- 12075640 TI - Implementation and evaluation of guidelines for preoperative testing in a tertiary hospital. AB - The aim of this study was to determine the extent of unnecessary investigation performed as part of the preoperative preparation of elective surgical patients in a teaching hospital and to audit the effect of guidelines and education designed to reduce unnecessary investigation. Guidelines were developed for preoperative anaesthetic investigation for elective surgical procedures in patients over one year of age according to internationally accepted criteria, with some adjustment for local differences in patient morbidity. Forrms outlining these criteria were placed in all operating theatres and anaesthetists were asked to determine whether tests performed were indicated or not, according to these criteria, over a two-week period, in each patient undergoing elective surgery. Tests indicated for surgical reasons were excluded. These same guidelines were then issued to all surgical departments along with explanatory lectures. The audit was repeated six months later and results compared. The incidence of over investigation decreased from 13.8% to 11.6% (P = 0.03) without a significant increase in under-investigation (0.7 v 1.0%; P = 0.2). This study highlights the incidence of unwarranted screening tests in patients presenting for elective surgery and the role of protocols and ongoing education in reducing this incidence. PMID- 12075641 TI - Ropivacaine 7.5 mg/ml versus bupivacaine 5 mg/ml for interscalene brachial plexus block--a comparative study. AB - We investigated ropivacaine 75 mg/ml in comparison with bupivacaine 5 mg/ml in patients receiving interscalene brachial plexus block (ISB) and general anaesthesia. In this randomized, double-blind, prospective clinical trial, each patient received an ISB block according to the technique originally described by Winnie and a catheter technique as per Meier. The rapidity of onset and the quality of sensory and motor block were determined. After general anaesthesia had been induced further parameters evaluated were consumption of local anaesthetic, opioid and neuromuscular blocking drug. After arrival in the recovery room, the patients were assessed for intensity of pain using a visual analog scale (VAS). One hundred and twenty patients were included in the study. The onset and development of sensory block was similar in both groups. Development and quality of motor block was also nearly identical for both local anaesthetics. Consumption of neuromuscular blocking drug and opioid did not differ between ropivacaine and bupivacaine. In the recovery room the mean pain score was less than 25 in both groups. There were no significant differences in terms of onset and quality of sensory or motor block during the intraoperative and early postoperative period. In addition we did not identify any side-effects related to the administration of the local anaesthetics. Ropivacaine 7.5 mg/ml and bupivacaine 5mg/ml proved to be nearly indistinguishable when administered for interscalene brachial plexus block. PMID- 12075642 TI - Washing of gloved hands in antiseptic solution prior to central venous line insertion reduces contamination. AB - Glove contamination at the time a central venous catheter is handled is highly undesirable and likely to increase the risk of subsequent line infection. This study was designed to determine how frequently gloves become contaminated during central venous line insertion and to demonstrate the value of glove decontamination immediately prior to handling of the central venous catheter During twenty routine internal jugular catheter insertions the sterility of the operator's gloved fingertips (just prior to handling the intravenous catheter) was assessed by touching the fingertips onto blood agar plates. The gloved hands were then rinsed in chlorhexidine/alcohol and after drying were placed onto a further plate. Contamination was detected in 55% of the prewash plates but in none of the postwash plates. Procedures performed by less experienced resident staff had a higher contamination rate despite there being no evident breach of sterile technique. It is likely that glove contamination results from the persistance of bacteria within the deeper layers of the skin, despite surface disinfection. These bacteria may be released by manipulation of the skin when identifying landmarks. This hypothesis was supported by a subsequent observation that gloves were more highly contaminated after firm touching of the skin rather than light touching. Glove contamination during central line insertion is frequent. Catheter contamination rates could be reduced (without risk or additional cost) by rinsing gloved hands in a solution of chlorhexidine (0.5%) in alcohol (70%) prior to handling the catheter. PMID- 12075643 TI - Variability in infusion pressure and continuous flow rate delivered from pressurized bag pump flush systems. AB - Using 10 different infusion bag pressure pumps, indicated manometer pressures were compared with measured infusion pressures proximal to the flow-regulating device in an in vitro experiment. Flow rates delivered through the flow regulating device were gravimetrically measured at different monitored pressure levels. Significant differences were found between manometer and measured infusion pressures among the tested pressure bagpumps (e.g. 500 ml bag volumepressurized to 300 mmHg manometerpressure: 219.6 +/- 7.8 to 407.2 +/- 2.7 mmHg). The infusion pressures were additionally affected by the vertical level of the infusion bag pump and by the volume of the infusion bag. Flow rates delivered through the flow-regulating device were directly correlated to the measured infusion pressure (r2= 0.9926). Differences inflow rates can have a considerable impact on maintaining catheter patency and avoidance of fluid overload and retrograde flushing into the central arterial circulation in neonates and small children. A simple manoeuvre using the invasive pressure transducer allows monitoring and adjustment of the infusion pressure in the clinical setting. PMID- 12075644 TI - High dependency units: issues to consider in their planning. AB - This review discusses the issues to be considered in establishing new or extending existing high dependency unit (HDU) services. A defined high dependency service becomes cost-effective when patient care requires more than one nurse for three patients. Professional guidelines for HDUs vary and there are no national accreditation criteria. Casemix and service delivery specifications for the HDU need to be defined and agreed upon within the institution. Establishing a new HDU service requires changes to care delivery. Many potential HDU patients are currently managed in general wards or in the intensive care unit. The service should be discussed widely and marketed within the institution, and the development of defined working relationships with the ICU and primary care teams on the wards is mandatory. PMID- 12075645 TI - Career choice influences in Australian anaesthetists. AB - All female members and a randomly selected group of male members of the Australian Society of Anaesthetists (n = 488) were surveyed by questionnaire as part of a broader study of gender issues in anaesthesia. This paper reports on reasons for career choice and the importance of role models. Responses were received from 199 women and 98 men (60.9% of those surveyed), representing all States and one Territory. Most males (95.9%) and a majority of females (55.7%) worked full-time. Reasons for career choice varied with gender, with a significantly greater proportion of women (39.7%) than men (8.7%) choosing anaesthesia because of controllable hours, particularly the ability to work part time. Experiences in anaesthesia during internship and residency were important for 19.1% of women and 14.1% of men, although very few mentioned undergraduate exposure. Other important factors in career choice were the application of physiology and pharmacology in patient care, practical and procedural aspects of practice, and chance. A majority of women (56%) and men (55%) named specific role models who were influential and encouraging in their choice. These results are similar to those of other studies. PMID- 12075646 TI - Prolonged paralysis following mivacurium administration. AB - Mivacurium is a benzylisoquinolone, choline-like, non-depolarizing muscle relaxant. Its onset of action is similar to that of atracurium but its duration of action is shorter (approximately 10-15 minutes). Mivacurium is metabolized by plasma cholinesterases at approximately 70% of the rate of metabolism of suxamethonium. Deficiency or abnormality of plasma cholinesterase may cause the duration of action of both suxamethonium and mivacurium to be greatly prolonged. We describe a case of prolonged mivacurium paralysis after day surgery. Laboratory investigations showed a genetic tendency toward abnormal cholinesterase levels, but markedly depressed cholinesterase activity was suggestive of additional acquired causes. This patient had a history of liver disease, malnutrition and anticholinesterase use, which we believe were the most significant factors involved. PMID- 12075647 TI - Vancomycin reaction during spinal anaesthesia. AB - A 77-year-old female received uneventful spinal anaesthesia for a total knee replacement. Upon the advice of the microbiologists and at the request of the orthopaedic surgeon, a vancomycin infusion was commenced prior to the application of the leg tourniquet. Five minutes later, having received only 40 mg of the antibiotic dose, she became unconscious and suffered severe cardiovascular collapse, from which she was resuscitated with intravenous ephedrine and adrenaline. PMID- 12075648 TI - Intra-arterial injection of metoclopramide, midazolam, propofol and pethidine. AB - A case is reported in which metoclopramide, midazolam, propofol, and pethidine were injected arterially during the course of anaesthesia, resulting in oedema and discolouration of the hand initially, but no long-term sequelae. An AVI 470 pump (3M Corporation) was used to control the infusion, andprevented backflow through a cannula which was unintentionally sited in an artery rather than a vein. Few anaesthetic drugs have their effects well documented when given arterially, and in particular, there were no previous reported cases involving metoclopramide in this context. The pharmacology of metoclopramide is discussed with reference to the drug's known effects on vascular beds and platelet function. PMID- 12075649 TI - Excruciating perineal pain after intravenous dexamethasone. AB - We describe two cases of unexpected perineal pain immediately after intravenous injection of fentanyl and dexamethasone (100 microg and 8 mg respectively) during induction of general anaesthesia. In both cases the pain was immediate (onset within 30 seconds), severe, localized to the genital region and of shooting and burning character No other clinical signs or symptoms were observed in either case and both patients made an uneventful recovery without neurological sequelae. We review the existing literature on perineal pain as an adverse effect of intravenous corticosteroid esters and recommend their administration either in diluted form or after induction of general anaesthesia. PMID- 12075650 TI - Anaesthetic management of a patient with Emery-Dreifuss muscular dystrophy. AB - Emery-Dreifuss muscular dystrophy is a rare form of muscular dystrophy associated with cardiac implications such as cardiomyopathy and arrhythmias leading to sudden death. We describe the anesthetic management of a patient with Emery Dreifuss muscular dystrophy who presented for orthopaedic surgery and discuss the disorder and its potential anaesthetic implications. PMID- 12075651 TI - Total intravenous anaesthesia for tracheobronchial stenting in children. AB - Stenosis and malacia of the tracheobronchial tree, most often secondary to prolonged intubation, tracheostomy or following correction of a congenital cardiac lesion, present a significant therapeutic problem, especially when the lesions are extensive. The utilization of self-expanding tracheobronchial stents is a useful addition to the medical armamentarium for maintenance of airways in these patients with major airway stenosis and collapse. The majority of previous reported cases of tracheobronchial stenting have been performed under general anaesthesia with the help of rigid bronchoscopy under direct vision. We conducted two cases of tracheobronchial stenting in postoperative cardiosurgical babies under continuous propofol infusion taking advantage of cardiovascular stability during continuous infusion and rapid emergence after its discontinuation. PMID- 12075652 TI - Ventricular tachycardia in congenital adrenal hyperplasia. AB - Severe hyperkalaemia in patients with congenital adrenal hyperplasia in association with aggravating factors such as acidosis and hypocalcaemia can cause life-threatening ventricular arrhythmias. Treatment of the underlying cause may be the only modality required in such cases. We report a 20-day-old male presenting with ventricular tachycardia due to electrolyte abnormalities in salt losing congenital adrenal hyperplasia. Sudden cardiac deaths reported earlier in such cases thus gain credence. PMID- 12075653 TI - Ketamine dependence. AB - Ketamine hydrochloride is a safe and rapid-acting non-opioid, lipid soluble anaesthetic with a short elimination half-life that is used for medical and veterinary purposes. It produces a state of "dissociative anaesthesia", probably from action on N-methyl-D-aspartate (NMDA) receptors. The psychotropic effects of ketamine range from dissociation and depersonalization to psychotic experiences and include a sensation of feeling light, body distortion, absence of time sense, novel experiences of cosmic oneness and out-of-body experiences. Abuse of ketamine has been reported, the typical abuser being an individual who uses multiple drugs and has some contact with medical agencies. This case demonstrates the effects of large doses of ketamine in a person with polysubstance abuse. The case also highlights development of significant tolerance to ketamine without prominent withdrawal symptoms. Caution in use of ketamine is reiterated in light of its abuse liability. PMID- 12075654 TI - Explosive separation of a Comweld oxygen regulator. PMID- 12075655 TI - Usage of digital cameras for preoperative dental evaluations especially in emergency anaesthesia. PMID- 12075656 TI - Surgical masks in the operating theatre: they are really necessary in cardiovascular procedures. PMID- 12075657 TI - Severe bradycardia in a patient with myasthenia gravis during transurethral ureterolithotripsic procedure under spinal anaesthesia. PMID- 12075658 TI - Surgical emphysema and dentoalveolar surgery. PMID- 12075659 TI - Transient cardiopulmonary arrest following retrobulbar block with lignocaine. PMID- 12075660 TI - Rofecoxib reduces perioperative morphine consumption for abdominal hysterectomy and laparoscopic gastric banding. PMID- 12075661 TI - Arthroscopic intubation. PMID- 12075662 TI - Protection against aspiration with the Proseal laryngeal mask airway. PMID- 12075663 TI - Partitioning the UMLS semantic network. AB - The unified medical language system (UMLS) integrates many well-established biomedical terminologies. The UMLS semantic network (SN) can help orient users to the vast knowledge content of the UMLS Metathesaurus (META) via its abstract conceptual view. However, the SN itself is large and complex and may still be difficult to comprehend. Our technique partitions the SN into smaller meaningful units amenable to display on limited-sized computer screens. The basis for the partitioning is the distribution of the relationships within the SN. Three rules are applied to transform the original partition into a second more cohesive partition. PMID- 12075664 TI - Evaluation and application of a semantic network partition. AB - Semantic networks (SNs) are excellent knowledge representation structures. However, large semantic networks are hard to comprehend. To overcome this difficulty, several methods of partitioning have been developed that rely on different mixes of structural and semantic methods. However, little has appeared in the literature concerning the question whether a partition of a semantic network creates subnetworks that agree with human insight. We address this issue by presenting a comparison between the results of an algorithmic partitioning method and a partition created by a group of experts. Subsequently, we show how a network partition can be used to generate various partial views of a semantic network, which facilitate user orientation. Examples from the Unified Medical Language System (UMLS) SN are used to demonstrate partial views. PMID- 12075665 TI - Architecture of a mediator for a bioinformatics database federation. AB - Developments in our ability to integrate and analyze data held in existing heterogeneous data resources can lead to an increase in our understanding of biological function at all levels. However, supporting ad hoc queries across multiple data resources and correlating data retrieved from these is still difficult. To address this, we are building a mediator based on the functional data model database, P/FDM, which integrates access to heterogeneous distributed biological databases. Our architecture makes use of the existing search capabilities and indexes of the underlying databases, without infringing on their autonomy. Central to our design philosophy is the use of schemas. We have adopted a federated architecture with a five-level schema, arising from the use of the ANSI-SPARC three-level schema to describe both the existing autonomous data resources and the mediator itself. We describe the use of mapping functions and list comprehensions in query splitting, producing execution plans, code generation, and result fusion. We give an example of cross-database querying involving data held locally in P/FDM systems and external data in SRS. PMID- 12075666 TI - Biological data integration: wrapping data and tools. AB - Nowadays scientific data is inevitably digital and stored in a wide variety of formats in heterogeneous systems. Scientists need to access an integrated view of remote or local heterogeneous data sources with advanced data accessing, analyzing, and visualization tools. Building a digital library for scientific data requires accessing and manipulating data extracted from flat files or databases, documents retrieved from the Web as well as data generated by software. We present an approach to wrapping web data sources, databases, flat files, or data generated by tools through a database view mechanism. Generally, a wrapper has two tasks: it first sends a query to the source to retrieve data and, second builds the expected output with respect to the virtual structure. Our wrappers are composed of a retrieval component based on an intermediate object view mechanism called search views mapping the source capabilities to attributes, and an eXtensible Markup Language (XML) engine, respectively, to perform these two tasks. The originality of the approach consists of: 1) a generic view mechanism to access seamlessly data sources with limited capabilities and 2) the ability to wrap data sources as well as the useful specific tools they may provide. Our approach has been developed and demonstrated as part of the multidatabase system supporting queries via uniform object protocol model (OPM) interfaces. PMID- 12075667 TI - OILing the way to machine understandable bioinformatics resources. AB - The complex questions and analyses posed by biologists, as well as the diverse data resources they develop, require the fusion of evidence from different, independently developed, and heterogeneous resources. The web, as an enabler for interoperability, has been an excellent mechanism for data publication and transportation. Successful exchange and integration of information, however, depends on a shared language for communication (a terminology) and a shared understanding of what the data means (an ontology). Without this kind of understanding, semantic heterogeneity remains a problem for both humans and machines. One means of dealing with heterogeneity in bioinformatics resources is through terminology founded upon an ontology. Bioinformatics resources tend to be rich in human readable and understandable annotation, with each resource using its own terminology. These resources are machine readable, but not machine understandable. Ontologies have a role in increasing this machine understanding, reducing the semantic heterogeneity between resources and thus promoting the flexible and reliable interoperation of bioinformatics resources. This paper describes a solution derived from the semantic web [a machine understandable world-wide web (WWW)], the ontology inference layer (OIL), as a solution for semantic bioinformatics resources. The nature of the heterogeneity problems are presented along with a description of how metadata from domain ontologies can be used to alleviate this problem. A companion paper in this issue gives an example of the development of a bio-ontology using OIL. PMID- 12075668 TI - Building a bioinformatics ontology using OIL. AB - This paper describes the initial stages of building an ontology of bioinformatics and molecular biology. The conceptualization is encoded using the ontology inference layer (OIL), a knowledge representation language that combines the modeling style of frame-based systems with the expressiveness and reasoning power of description logics (DLs). This paper is the second of a pair in this special issue. The first described the core of the OIL language and the need to use ontologies to deliver semantic bioinformatics resources. In this paper, the early stages of building an ontology component of a bioinformatics resource querying application are described. This ontology (TaO) holds the information about molecular biology represented in bioinformatics resources and the bioinformatics tasks performed over these resources. It, therefore, represents the metadata of the resources the application can query. It also manages the terminologies used in constructing the query plans used to retrieve instances from those external resources. The methodology used in this task capitalizes upon features of OIL-The conceptualization afforded by the frame-based view of OIL's syntax; the expressive power and reasoning of the logical formalism; and the ability to encode both handcrafted, hierarchies of concepts, as well as defining concepts in terms of their properties, which can then be used to establish a classification and infer relationships not encoded by the ontologist. This ability forms the basis of the methodology described here: For each portion of the TaO, a basic framework of concepts is asserted by the ontologist. Then, the properties of these concepts are defined by the ontologist and the logic's reasoning power used to reclassify and infer further relationships. This cycle of elaboration and refinement is iterated on each portion of the ontology until a satisfactory ontology has been created. PMID- 12075669 TI - White and black blood volumetric angiographic filtering: ellipsoidal scale-space approach. AB - Prefiltering is a critical step in three-dimensional (3-D) segmentation of the blood vessel and its display (see the recent book by Suri et al.). This paper presents a scale-space approach for filtering the white blood and black blood angiographic volumes and its implementation issues. The raw MR angiographic volume is first converted to isotropic volume followed by 3-D higher order separable Gaussian derivative convolution with known scales to generate edge volume. The edge volume is then run by the directional processor at each voxel where the eigenvalues of the 3-D ellipsoid are computed. The vessel score per voxel is then estimated based on these three eigenvalues which suppress the nonvasculature and background structures yielding the filtered volume. The filtered volume is ray-cast to generate the maximum intensity projection images for display. The performance of the system is evaluated by computing the mean, variance, signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) images. The system is run over 20 patient studies from different areas of the body such as the brain, abdomen, kidney, knee, and ankle. The computer program takes around 150 s of processing time per study for a data size of 512 x 512 x 194, which includes the complete performance evaluation. We also compare our strategy with the recently published MR filtering algorithms by Alexander et al. and Sun et al. PMID- 12075670 TI - Auditory stimulus optimization with feedback from fuzzy clustering of neuronal responses. AB - The primary focus of this paper was to develop a high-performance computer system for optimizing auditory stimuli based on neuronal feedback. Using the Algorithm Of Pattern EXtraction (ALOPEX) extra-cellular action potentials (APs) recorded from frog (Rana Pipiens) auditory neurons were used as feedback to optimize sound stimuli. This computer-based system works in real time to iteratively find the neuron's best excitatory frequency (BEF). Three programmable (positive and negative) threshold logic levels are used to collect 300 APs in response to normalized pure tones. Fuzzy logic is then used to separate up to five fuzzy centers (templates) from the 300 APs. The fuzzy centers are used for on-line fuzzy mapping of future responses. The five fuzzy centers allow the system to monitor up to five neighboring neurons. To study the auditory neurons of the frog, one, two, and three simultaneous tones are used as the stimulus for optimization of the best combination of frequencies. Testing with the response calculated as a parabolic function of a single best frequency demonstrated system dynamics and reliability for up to nine simultaneous tones. Experiments using one pure tone and ten stimulus presentations per iteration showed that the automated system is able to repeatedly converge to the best frequency within 100 iterations. Studies using one, two, and then three pure tones played simultaneously on the same group of neurons has shown that these tones converged on the same best frequencies by properly mixing the tones available to produce the optimal complex sound. PMID- 12075671 TI - Rapid automated three-dimensional tracing of neurons from confocal image stacks. AB - Algorithms are presented for fully automatic three-dimensional (3-D) tracing of neurons that are imaged by fluorescence confocal microscopy. Unlike previous voxel-based skeletonization methods, the present approach works by recursively following the neuronal topology, using a set of 4 x N2 directional kernels (e.g., N = 32), guided by a generalized 3-D cylinder model. This method extends our prior work on exploratory tracing of retinal vasculature to 3-D space. Since the centerlines are of primary interest, the 3-D extension can be accomplished by four rather than six sets of kernels. Additional modifications, such as dynamic adaptation of the correlation kernels, and adaptive step size estimation, were introduced for achieving robustness to photon noise, varying contrast, and apparent discontinuity and/or hollowness of structures. The end product is a labeling of all somas present, graph-theoretic representations of all dendritic/axonal structures, and image statistics such as soma volume and centroid, soma interconnectivity, the longest branch, and lengths of all graph branches originating from a soma. This method is able to work directly with unprocessed confocal images, without expensive deconvolution or other preprocessing. It is much faster that skeletonization, typically consuming less than a minute to trace a 70-MB image on a 500-MHz computer. These properties make it attractive for large-scale automated tissue studies that require rapid on-line image analysis, such as high-throughput neurobiology/angiogenesis assays, and initiatives such as the Human Brain Project. PMID- 12075672 TI - Extending ventilation duration estimations approach from adult to neonatal intensive care patients using artificial neural networks. AB - In earlier work, the research group successfully used artificial neural networks (ANNs) to estimate ventilation duration for adult intensive care unit (ICU) patients. The ANNs performed well in terms of correct classification rate (CCR) and average squared error (ASE) classifying the outcome into two classes: whether patients were ventilated for less than/equal to or for more than 8 h (< or >). The objective of new work was to apply this adult model to the estimation of ventilation with neonatal ICU (NICU) patient records. The performance obtained with the neonatal patients was comparable to that previously found with the adult database, again as measured in terms of a maximum CCR and a minimum ASE. The effectiveness of using the weight-elimination technique in controlling overfitting was again validated for the neonatal patients as it had been for our adult patients. It was concluded that the approach developed for ICU adult patients was also successfully applied to a different medical environment: neonatal ICU patients. PMID- 12075673 TI - Residential radon exposure and lung cancer: risk in nonsmokers. AB - Lung cancer is a disease that is almost entirely caused by smoking; hence, it is almost totally preventable. Yet there are a small percentage of cases, perhaps as many as 5 to 15%, where there are other causes. Risk factors identified for this other group include passive smoking, occupational exposure to certain chemicals and ionizing radiation, diet, and family history of cancer. In the United States cigarette smoking is on the decline among adults, occupational exposures are being reduced, and people are being made more aware of appropriate diets. These changes are gradually resulting in a reduced risk for this disease. Lung cancer in the U.S. may, therefore, eventually become largely a disease of the past. It remains important, however, to continue to study the cause(s) of lung cancer in non-smokers, particularly never smokers. Because of our interest in the effects of residential radon exposure on the development of lung cancer in non-smokers, we conducted a critical review of the scientific literature to evaluate this issue in detail. Strict criteria were utilized in selecting studies, which included being published in a peer reviewed journal, including non-smokers in the studied populations, having at least 100 cases, and being of case-control design. A total of 12 individual studies were found that met the criteria, with 10 providing some information on non-smokers. Most of these studies did not find any significant association between radon and lung cancer in non-smokers. Furthermore, data were not presented in sufficient detail for non-smokers in a number of studies. Based on the most recent findings, there is some evidence that radon may contribute to lung cancer risk in current smokers in high residential radon environments. The situation regarding the risk of lung cancer from radon in non-smokers (ex and never) is unclear, possibly because of both the relatively limited sample size of non-smokers and methodological limitations in most of the individual studies. A summary of these studies is provided concerning the state of knowledge of the lung cancer risk from radon, methodological problems with the residential studies, the need for the provision of additional data on non-smokers from researchers, and recommendations for future research in non-smokers. PMID- 12075674 TI - Airborne 222Rn concentration in an Egyptian village. AB - The indoor radon concentration in a typical Egyptian village in Delta was measured in 50 houses during summer 1999 and winter 2000. This survey was done using three different types of charcoal canisters: 4" open-faced, 2.75" open faced, and 2.75" diffusion barrier. The average winter to summer ratio was 2.1 and 1.2 for open rooms and closed rooms, respectively, in clay buildings, while it was 0.8 and 1.0, respectively, in concrete buildings. The seasonal variations of radon concentration as well as the effect of building materials were discussed. PMID- 12075675 TI - Isotopic ratio of 129I/127I in seaweed measured by neutron activation analysis with gamma-gamma coincidence. AB - 129I is a long-lived (1.6 x 10(7) y) radionuclide that is produced in nature as the result of spontaneous fission of heavy elements and reaction of xenon with cosmic rays. Recently, however, artificial sources from nuclear power plants and nuclear test explosions have become a significant component of environmental radioactive iodine. Coincidence gamma-ray detection using Ge detectors makes it possible to simultaneously resolve the numerous gamma-rays produced by neutron activation. In this study, the coincidence gamma-ray detection technique was combined with neutron activation analysis to determine the radioactive iodine composition of seaweed. The ratio of 129I/127I in this common Japanese food item collected from the Ibaraki prefecture has been derived without the need for radiochemical purification. The isotopic ratio of 129I/127I in Kajime algae is 3.5(5) x 10(-10). PMID- 12075676 TI - IAEA model project on upgrading the radiation protection infrastructure: background--origin--implementation. International Atomic Energy Agency Model. AB - The International Atomic Energy Agency Model Project is a focused management program to improve the Radiation Control Programs primarily in developing countries. The effort was initiated in 1994 as a result of RAPAT mission reports and incidents around the world involving injuries and some deaths from uncontrolled and misuse of radioactive sources. This indicated that many countries were not implementing the IAEA Basic Safety Standards. The initial phase covered 52 countries and resulted in 69% passing legislation and 75% had some type of regulatory authority and 40% had regulations and licensing and inspection capabilities. But, the overall averages masked disappointment for some countries in West Asia where only 22% had regulations and 33% had a licensing and inspection program. In Africa 24% had regulations and 29% had a licensing and inspection program. Plans were approved to extend the effort to an additional 30 countries over the next 4 y. PMID- 12075677 TI - The international atom: evolution of radiation control programs. AB - Under the Atoms for Peace program, Turkey received a one MWt swimming pool reactor in 1962 that initiated a health physics program for the reactor and a Radiation Control Program (RCP) for the country's use of ionizing radiation. Today, over 13,000 radiation workers, concentrated in the medical field, provide improved medical care with 6,200 x-ray units, including 494 CAT scanners, 222 radioimmunoassay (RIA) labs and 42 radiotherapy centers. Industry has a large stake in the safe use of ionizing radiation with over 1,200 x-ray and gamma radiography and fluoroscopic units, 2,500 gauges in automated process control and five irradiators. A 48-person RCP staff oversees this expanded radiation use. One incident involving a spent 3.3 TBq (88 Ci) 60Co source resulted in 10 overexposures but no fatalities. Taiwan received a 1.6 MWt swimming pool reactor in 1961 and rapidly applied nuclear technology to the medical and industrial fields. Today, there are approximately 24,000 licensed radiation workers in nuclear power field, industry, medicine and academia. Four BWRs and two PWRs supply about 25% of the island's electrical power needs. One traumatic event galvanized the RCP when an undetermined amount of 60Co was accidentally incorporated into reinforcing bars, which in turn were incorporated into residential and commercial buildings. Public exposures were estimated to range up to 15 mSv (1.3 rem) per annum. There were no reported ill effects, except possibly psychological, to date. The RCP now has instituted stringent control measures to ensure radiation-free dwellings and work places. Albania's RCP is described as it evolved since 1972. Regulations were promulgated which followed the IAEA Basic Safety Standards of that era. With 525 licenses and 600 radiation workers, the problem was not in the regulations per se but in their enforcement. The IAEA helped to upgrade the RCP as the economy evolved from one that was centrally planned economy to a free market economy. As this transition takes place, public radiation exposures in the medical field will continue to be high until the old x-ray equipment is phased out. A small conscientious health physics staff works with limited resources to keep radiation exposures at acceptable levels. These three country RCPs, as they have evolved, have some commonality. Today, all radiation installations are licensed, both for radioactive material and x-ray equipment. Radiation workers are individually licensed or registered. All RCPs have, or are striving to have, their radiation regulations conform to ICRP 60 recommendations as spelled out in the Basic Safety Standard (1996). Finally, all three countries have as yet to find a permanent solution for their radioactive waste. PMID- 12075678 TI - New UK population medical exposure statistics. PMID- 12075679 TI - NRC revises regulations on medical uses of radioactive material. Nuclear Regulatory Commission. PMID- 12075680 TI - Reconstruction of two victims' posturing based on the induced radioactivities in their bones in the criticality accident in Tokai-Mura, Japan. AB - On 30 September 1999, three workers were severely exposed to neutrons and gamma rays in a criticality accident that occurred at a uranium conversion facility in Tokai-mura, Ibaraki Prefecture, Japan. Radiochemical analyses of 32P and 45Ca induced by neutrons in bone matrix were carried out after the deaths of two of the victims. It was found that more than several million becquerels of both nuclides had been produced in their body skeletons. Results showed non homogeneous distributions of neutron fluence in the bodies, from which it could be deduced how both workers were positioned relative to the fission source during exposure, i.e., at the moment of the first nuclear excursion. For the victim who died first, the activities in the central part of his body were more than those of his extremities. Also, in the central part of his body, the right side showed more activities than the left side. As for the second man, the activities indicated rather uniform exposure to neutrons to the whole body although the geometrical distribution of the activity varied enough to assume his orientation. Such information on the geometrical distribution of neutron-induced radioactivities in the skeleton can be used to reconstruct the posturing of the victims, which is necessary to estimate their apparent absorbed doses. PMID- 12075681 TI - Characteristics of the German uranium miners cohort study. AB - To evaluate the risk of cancer associated with low and high levels of radon exposure one of the largest single cohort studies on uranium miners is being conducted in Germany including 58,721 men who were employed for at least 6 mo between 1946 and 1989 at the former Wismut uranium company in Eastern Germany. Information on job history, smoking, dust, and arsenic was collected from the original payrolls and the medical records. Exposure to radon and its progeny was estimated by using a detailed job-exposure matrix. The first mortality follow-up determining the vital status as of 31 December 1998 has been started. The cohort includes 49,342 exposed miners who have worked underground or in processing/milling facilities and 9,379 never-exposed workers. Miners who had been exposed for the first time between 1946 and 1954 (n = 19,865), the years with the poorest working conditions, show higher mean cumulative radon exposures (709 working level months, WLM) and a longer duration of exposure (mean = 13 y) than those with the first exposure between 1955 to 1970 (121 WLM and 11 y, n = 14,155) or after 1970 (10 WLM and 6 y, n = 15,322), respectively. Information on smoking is available for 38% of the cohort, demonstrating that most miners were heavy smokers. In the first mortality follow-up a total of about 15,000 deceased men including about 2,200 lung cancer deaths are expected. The main strengths of the study are its size and the large group of workers having received low exposures over relatively long periods of time. PMID- 12075682 TI - Gastrointestinal absorption of uranium in humans. AB - The gastrointestinal absorption factor (f1) for uranium in humans has been determined from a study of 50 volunteers, ingesting uranium at natural levels in drinking water and food. The purpose of the study was to find an appropriate f1 value for humans to use in deriving exposure guidelines for uranium. The participants ranged in age from 13 to 87 years. They were selected from two communities: New Ross, Nova Scotia with elevated uranium in drinking water, and Ottawa, Ontario with very low levels of uranium. Uranium intake and excretion were measured in samples collected concurrently from the same individuals over a three-day period. The duplicate diet method was used to monitor uranium intake in food and water. Uranium levels in all samples were measured by inductively coupled plasma mass spectrometry (ICP/MS). The distribution of f1 values obtained was non-Gaussian with a range of 0.001 to 0.06 and a median of 0.009. Seventy eight percent of the subjects had values less than 0.02. These values are consistent with the recommendations of ICRP 69. The f1 values were not gender sensitive and were independent of age at time of study, duration of exposure, and total uranium intake. The implications of these findings are discussed in terms of setting drinking water guidelines. PMID- 12075683 TI - Nation-wide survey on radiation doses in diagnostic and interventional radiology in Switzerland in 1998. AB - A nation-wide survey on radiation doses in diagnostic and interventional radiology was conducted in Switzerland in 1998 aiming at establishing their collective radiological impact on the Swiss population. The study consisted on the one hand of surveying the frequency of more than 250 types of examinations, covering conventional radiology, mammography, fluoroscopy, angiography, interventional radiology, CT, bone densitometry, conventional tomography and dental radiology. On the other hand, for each type of examination the associated patient dose was established by modeling. The results of this study show that about 9.5 million diagnostic and interventional examinations are performed annually in Switzerland (1.34 per caput) and that the associated annual collective dose is of the order of 7100 person.Sv (1.0 mSv per caput). Switzerland is similar to other European countries in terms of the frequency of examinations and the collective dose. PMID- 12075684 TI - Absorption of strontium from the gastrointestinal tract into plasma in healthy human adults. AB - The radioactive isotopes of strontium have always been a major concern in radiation protection. Currently, radiostrontium is of interest for evaluation of the health effects of the Chernobyl accident and for epidemiological studies in populations exposed to releases from the Mayak nuclear facilities in Russia. Ingestion is one of the most important exposure pathways involving radioactive strontium. The main sources of published data on the fraction of the ingested strontium that is transferred to plasma (f1) are summarized. For some of these studies, the original data had to be reanalyzed and a new iterative method to account for the elimination in feces of strontium of endogenous origin (i.e., that was absorbed to blood and has already been returned into feces) was employed. Data indicate no significant dependence of the absorbed fraction on sex or age at exposure within the adult group, but absorption of strontium is reduced if the intake of stable calcium is very high and is enhanced if the intake of calcium is very low. The probability distribution function of f1 values is well represented by a lognormal curve with a geometric mean of 22.3% and a geometric standard deviation of 1.44 (95% confidence interval 10.9% to 45.6%, or about a factor of 2 around the geometric mean). This distribution can be considered representative for the variability of the f1 values in a population of healthy adults. PMID- 12075685 TI - Patient effective radiation dose and associated risk from transmission scans using 153Gd line sources in cardiac spect studies. AB - The aim of the present study was to determine the contribution of transmission measurements acquisition to total patient effective dose from cardiac SPECT studies. A dual-head L-shaped gamma camera equipped with a transmission scan acquisition system based on two 153Gd line sources was used to simulate transmission measurements acquisition exposure on an anthropomorphic phantom. Thermoluminescence dosimeters were used to directly monitor the dose to 550 measuring points in the phantom. The effective dose and associated risk from transmission scans acquisition were estimated and compared to those associated to the radiopharmaceutical injected. The maximum effective dose from a typical transmission measurements acquisition was 1.3 microSv and 1.9 microSv for male and female patients, respectively. The contribution of the typical transmission scans acquisition to total patient radiation risk from a cardiac SPECT study is less than 10(-3). Thus, radiation exposure may not be considered as a limiting factor for the clinical application of attenuation correction methods based on transmission measurements in cardiac SPECT. PMID- 12075686 TI - An EPR model for separating internal 90Sr doses from external gamma-ray doses in teeth. AB - The problem of retrospectively measuring radiation doses to humans having both external and internal exposures is addressed. A simplified theoretical model is presented by which the internal exposure from a bone seeking radionuclide can be measured and distinguished from an external gamma-ray dose. The model also provides a means of estimating initial acute uptakes of 90Sr. PMID- 12075687 TI - Millimeter wave absorption in the nonhuman primate eye at 35 GHz and 94 GHz. AB - The purpose of this study was to evaluate anterior segment bioeffects of pulsed 35 GHz and 94 GHz microwave exposure in the nonhuman primate eye. Five juvenile rhesus monkeys (Macaca mulatta) underwent baseline anterior segment ocular assessment consisting of slit lamp examination, corneal topography, specular microscopy, and pachymetry. These studies were repeated after exposure of one eye to pulsed 35 GHz or 94 GHz microwaves at varied fluences, with the other eye serving as a control. The mean fluence required to produce a threshold corneal lesion (faint epithelial edema and fluorescein staining) was 7.5 J cm(-2) at 35 GHz and 5 J cm(-2) at 94 GHz. Transient changes in corneal topography and pachymetry were noted at these fluences. Endothelial cell counts remained unchanged. Threshold corneal injury from 35 GHz and 94 GHz microwave exposure is produced at fluences below those previously reported for CO2 laser radiation. These data may help elucidate the mechanism of thermal injury to the cornea, and resolve discrepancies between IEEE C95.1 (1999), NCRP (1986), and ICNIRP (1998) safety standards for exposure to non-ionizing radiation at millimeter wavelengths. PMID- 12075688 TI - Diurnal radon variations in the upper soil layers and at the soil-air interface related to meteorological parameters. AB - Measurements of the radon concentration in a column (1 m2 x 2 m) consisting of a homogeneous mixture of dry sand and uranium tailings have been performed to obtain information on the radon transport under well defined conditions. The dependence of the radon concentration has been exclusively studied on the soil/air temperature gradient and on the wind speed. The soil moisture content has been kept constant. Significant diurnal variations of the radon concentration were detected in the uppermost soil layer and at the soil/air interface. Such a behavior was not found in 30 cm and deeper soil layers. It is argued that the diurnal radon variation in the uppermost soil layer is mainly associated with the diurnal inversion of the soil/air temperature gradient giving rise to a convective soil gas migration additional to the common upward diffusion processes, whereas the diurnal variation of the radon concentration at the soil/air interface is caused by the interplay of the temperature gradient and the wind speed. No impact of atmospheric pressure variations on the radon migration has been observed. PMID- 12075689 TI - Temperature and humidity consideration for calculating airborne 222Rn using activated charcoal canisters. AB - A radon calibration chamber with variable and controlled radon concentration, humidity, and temperature was constructed and used to extend the calibration range of open-faced and diffusion-barrier charcoal canisters for higher temperature and humidity applications. The optimum exposure time to achieve the best detection accuracy and the lowest minimum detectable level was between 2 and 4 d depending on relative humidity for open-faced canister and 4 d for diffusion barrier canister over a range of humidity. The dependence of radon adsorption on the temperature was also studied for both types of canister at different values of relative humidity. The open-faced canisters at 15 degrees C absorb 68% more radon than at 30 degrees C over the range of humidity from 20% to 80%. The diffusion-barrier canister shows insignificant variation in absorption over the same range of temperature and humidity. PMID- 12075690 TI - The power integration diffusion model for production breaks. AB - The learning curve during repetitive production and the associated forgetting during production breaks are fundamental issues in the understanding of behavior. A model is suggested that combines 3 basic findings, namely, that single memory traces decay according to a power function of the retention interval, that aggregated memory traces can be combined by integration, and that the time to produce a unit can be described by a diffusion process on the memory trace. This power integration diffusion model is validated with empirical data, and the result fits better than 14 other published forgetting models. PMID- 12075691 TI - New evidence on the suggestibility of memory: the role of retrieval-induced forgetting in misinformation effects. AB - Extending recent work that has demonstrated that the act of remembering can result in the inhibition of related items in memory, the present research examined whether retrieval-induced forgetting could provide a mechanism for explaining misinformation effects. Specifically, the authors found in their first study that the inhibition of critical items rendered the recollection of postevent information more likely in a subsequent test of memory. The authors established in their second study that when guided retrieval practice and final recall tests were separated by 24 hr, retrieval-induced forgetting failed to emerge and misinformation effects were absent. In contrast, a delay of 24 hr between initial encoding and guided retrieval practice produced not only retrieval-induced forgetting but also misinformation effects. PMID- 12075692 TI - Evaluation of a behavioral measure of risk taking: the Balloon Analogue Risk Task (BART). AB - The present study (N = 86) sought to evaluate a laboratory-based behavioral measure of risk taking (the Balloon Analogue Risk Task; BART) and to test associations between this measure and self-report measures of risk-related constructs as well as self-reported real-world risk behaviors. The BART evidenced sound experimental properties, and riskiness on the BART was correlated with scores on measures of sensation seeking, impulsivity, and deficiencies in behavioral constraint. Also, riskiness on the BART was correlated with the self reported occurrence of addictive, health, and safety risk behaviors, with the task accounting for variance in these behaviors beyond that accounted for by demographics and self-report measures of risk-related constructs. These results indicate that the BART may be a useful tool in the assessment of risk taking. PMID- 12075693 TI - Multimedia comprehension skill predicts differential outcomes of web-based and lecture courses. AB - College students (134 women and 55 men) participated in introductory psychology courses that were offered largely online (on the World Wide Web) or in a lecture format. Student comprehension skills were inferred from their scores on a multimedia comprehension battery. The learning of content knowledge was affected interactively by comprehension skill level and course format. Differences between format increased with comprehension skill such that the Web-based course advantage became greater as comprehension skill increased. This same pattern was not seen when self-reports of comprehension ability were used as the predictor. Furthermore, comprehension skill did not predict course satisfaction. Generally, students of all skill levels preferred the lecture courses. PMID- 12075694 TI - Investigating spatial vision and dynamic attentional selection using a gaze contingent multiresolutional display. AB - This study examined spatial vision and attentional selection using a gaze contingent multiresolutional display, with a dynamic, gaze-centered, high resolution window and lower resolution periphery. Visual search times and eye movements from 15 participants in a 3 x 3 design (Window Radius x Peripheral Resolution) suggest that contrast sensitivity as a function of retinal eccentricity affects attentional selection and visual processing. Smaller windows led to longer search times and shorter saccades; lower peripheral resolution also shortened saccades (all ps < .05) as a result of avoiding fixating degraded areas. Fixation durations, although longer for smaller windows (p < .05), were unaffected by whether the next saccade went within or outside the window. These results are explained through (a) competition among potential saccade targets where above-threshold filtering reduces an object's relative salience and (b) generally disrupted visual processing. PMID- 12075695 TI - New approaches to the imaging, diagnosis, and biopsy of breast lesions. AB - Currently, breast cancer is detected and diagnosed by a combination of physical examination, imaging, and biopsy. Particularly for younger women, the limitations of these methods can cause a cancerous lesion to go undetected because neither physical examination nor imaging are infallible, and neither can promise the early detection of all breast cancers. Dense breasts make physical examination, mammography, and ultrasound less sensitive. Breast screening of all types generates over a million breast biopsies in the U.S. alone. This article reviews how to optimize the approach to breast biopsy using available methods to minimize the number of procedures a woman experiences, and describes tools in development that will hopefully improve our ability to diagnose and treat breast cancer. PMID- 12075696 TI - Magnetic resonance image-guided focused ultrasound surgery. AB - The powerful union of focused ultrasound and magnetic resonance imaging (MRI) has created a new approach to noninvasive surgery. By using this integrated therapy delivery system, the physician can correctly localize tumors, optimally target acoustic energy, monitor energy deposition in real time, and accurately control the deposited thermal dose within the entire tumor volume. This satisfies the requirements for "ideal surgery." In a real sense, MRI provides the "road map" by which focused ultrasound surgery (FUS) is followed. The advantages of MRI over ultrasound guidance in controlling FUS lie in the more sensitive detection of tumor target, the real-time detection of tissue temperature, and the confirmation of thermally induced tissue changes-powerful features that eventually can replace the traditional surgical approach. Applying software that connects the therapy and imaging system (the "Dosimetry Workstation"), the physician can generate an entire treatment plan from quantifying temperature changes to positioning the therapy transducer. The noninvasive debulking of tumors without disturbing adjacent, functionally intact structures is thereby accomplished. PMID- 12075697 TI - Interventional endosonography. AB - Endoscopic ultrasound (EUS) has become the most accurate imaging modality for locoregional cancer staging of the gastrointestinal (GI) tract. Fine-needle aspiration (FNA) capabilities have added a whole new level of accuracy in nodal staging with reported numbers in the 90% range for luminal and pancreaticobiliary disease. In addition, new non-GI applications are being evaluated like the role of EUS-FNA for non-small cell lung carcinoma and exploration of the posterior mediastinum. Furthermore, the same capabilities that allow for safe tissue sampling are being explored for interventional applications like EUS-guided celiac plexus neurolysis and fine-needle injection. The following review describes the current clinical status of EUS in GI oncology as well as future and novel indications and therapeutic strategies for this technology. PMID- 12075698 TI - Lymphatic mapping and sentinel lymph node biopsy for breast cancer. AB - Lymphatic mapping and sentinel lymph node mapping have changed the standard of surgical care for patients with melanoma and have the potential to do the same for women with breast cancer. The technique will result in more accurate disease staging and in less morbidity. Preoperative lymphoscintigraphy and the intraoperative mapping technique are being standardized across the country. With the lymphatic mapping technique, approximately 10% of the so-called histologic negative population will be more accurately staged. National trials are ongoing to determine the clinical significance of the missed micrometastatic PMID- 12075699 TI - Update on the management of ovarian cancer. AB - Ovarian cancer remains the leading gynecologic cause of death in the United States and the Western world. Progression to metastatic disease prior to diagnosis contributes to the high mortality rate associated with ovarian cancer. The current article reviews surgical and drug therapies for ovarian cancer. Prognostic factors and preventative treatment are also discussed. Surgery is essential for accurate staging of ovarian cancer and treatment. Cytoreduction, combined with chemotherapy, may relieve symptoms associated with bowel obstruction and improve survival. Management of early-stage ovarian cancer depends upon risk status determined via comprehensive staging at the time of surgical resection. High-risk, but not low-risk, patients require adjuvant chemotherapy. Studies comparing various combinations of cytotoxic agents for the treatment of advanced stage ovarian cancer are described. Despite surgery and chemotherapy, ovarian cancer recurs in approximately 50% of patients. Management of recurrent ovarian cancer and maintenance therapy following remission are discussed. PMID- 12075700 TI - Update in the management of endometrial cancer. AB - Endometrial cancer is the most common malignancy of the female genital tract and the fourth most common malignancy in women. Standard treatment for early-stage disease involves surgical staging with adjuvant therapy based on pathologic findings. Controversial issues include the role of laparoscopy and the indications for lymph node sampling. Advanced and recurrent disease can be treated with surgery, chemotherapy, or radiation therapy based on the individual pattern of spread and prior treatment regimens. This review will begin by discussing the surgical treatment of early-stage endometrial cancer and the role of adjuvant radiation therapy. The role of surgery and chemotherapy in advanced disease will also be covered. Current experimental protocols and areas of future investigation will be discussed briefly throughout. The standard treatment of endometrial cancer has been the subject of many reviews in the literature. This paper will focus on areas of controversy in the management of endometrial cancer as well as new developments in the treatment of this disease. Recent published reports will be used to highlight changes to traditional practices. The reviewwill be limited to epithelial adenocarcinoma of the endometrium. PMID- 12075701 TI - Molecular pathogenesis of lung cancer and potential translational applications. AB - Molecular studies of lung cancer using individual genes and global approaches of gene analysis have shown several observations that are ready to be translated into clinically useful information to provide new methods of diagnosis, risk assessment, prevention, and treatment. Clinically evident lung cancers have acquired 20 or more clonal genetic alterations, and tumor acquired promoter hypermethylation is a frequent epigenetic mechanism of inactivation of gene expression in lung cancer giving at least another 10-20 lesions. Furthermore, small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) have acquired different genetic and epigenetic lesions. Alterations in 3p tumor suppression genes (TSGs) appear especially early, including those of RASSF1A and SEMA3B at 3p21.3, followed by changes in 9p (p16), 8p, and then multiple other sites. Changes consistent with oxidative damage leading to mitotic recombination are frequently seen. Smoking-damaged, histologically normal epithelium as well as epithelium with preneoplastic/preinvasive changes have thousands of clonal patches containing genetic alterations. Finally, correcting even single genetic abnormalities can reverse the malignant phenotype. PMID- 12075702 TI - CT screening for lung cancer: the first ten years. AB - This article reviews the current state of knowledge regarding computed tomography (CT) screening for lung cancer. The initial 3 studies have demonstrated the superiority of CT-based screening for lung cancer over traditional radiography. Furthermore, false-positive tests common on baseline screening were manageable without notable excess of biopsies or thoracotomies. Differences and similarities of various screening regimens are discussed in relation to reported study results. When comparing the various studies, the focus should be placed on the diagnostic distribution achieved under annual repeat screening, as it is this distribution which is obtained year after year of repeat screening and this distribution and the curability of these repeat screen-diagnosed lung cancer which determine the usefulness of the screening regimen. To date, these studies demonstrate a consistent shift to over 80% of the cancer being diagnosed in Stage IA. This marked shift suggest that the curability of screen-detected lung cancers will be markedly improved. PMID- 12075703 TI - Combined modality therapy for non-small cell lung carcinoma. AB - Combined modality therapy is frequently used to treat non-small cell lung carcinoma (NSCLC). Surgical resection is the mainstay of treatment for Stages I and II NSCLC. Little evidence supports the use of adjuvant therapy In patients with Stage I NSCLC. Adjuvant radiotherapy to the chest after resection for Stage II and IIIA NSCLC remains controversial but may be appropriate for many patients with resected Stage IIIA disease or inadequately pathologically staged II disease. For Stage IIIA NSCLC, surgery appears suboptimal, though no other single modality appears superior. Recent studies of neoadjuvant therapy for Stage IIIA NSCLC yielded promising results. The standard of care for Stage III unresectable NSCLC and strategies to improve local-regional control in Stage III NSCLC are discussed, as are the importance of toxicity protection with chemoradiation and the potential utility of prophylactic cranial irradiation for preventing brain relapse. Targeted therapies appear promising, and expedited evaluation in Stage III NSCLC is warranted. PMID- 12075704 TI - Surgical approaches to liver metastases. AB - Metastases confined to the liver from tumors arising in the gastrointestinal tract or other sites represent a significant clinical problem. Although the majority of patients present with disease that is not amenable to resection based on the size, number, or anatomic distribution of tumors, for selected patients with limited extent of disease surgical resection can result in long-term survival. In patients with colorectal cancer, a number of adverse prognostic factors have been identified that are associated with shortened survival following hepatic resection. The role of adjuvant hepatic artery infusion and systemic chemotherapyfollowing resection in this patient population has been the topic of several prospective random assignment trials. For patients with unresectable metastases confined to liver, a number of regional therapies are in clinical development and share the advantages of intensifying therapy at the site of tumor while minimizing or eliminating unnecessary systemic exposure and toxicity. One such approach is vascular isolation and perfusion, also known as isolated hepatic perfusion or IHP, in which the liver is treated with high dose chemotherapy and/or biological agents under hyperthermic conditions. Although only a limited number of centers have reported substantial experience with this procedure, it appears to have significant efficacy even in patients with advanced tumor burden orthose with tumors refractory to other types of therapy. The status of IHP is presented. PMID- 12075705 TI - Hepatic artery infusion of chemotherapy as a treatment for hepatic metastases from colorectal cancer. AB - Hepatic artery infusion (HAI) of chemotherapy as a treatment for hepatic metastases from colorectal cancer has become more commonly used after the introduction of the totally implantable hepatic artery pump in the early 1970s. Floxuridine (FUDR) is the generally used chemotherapy agent in the pump because of its high solubility and high extraction rates by the liver on the first pass of the chemotherapy through the hepatic circulation. HAI has been used mainly to treat unresectable liver metastases in patients who have liver metastases only. The other scenario for pump use has been as an adjuvant therapy after resection of all metastatic disease inthe liver. The rationale for HAI includes the unique dual blood supply of the liver allowing chemotherapy given into the artery and sparing the normal cells, which get their predominant blood supply from the portal vein. The details of pump design will be reviewed. Complications from HAI are specific for this therapy and will be reviewed. Treatment of unresectable liver metastases with HAI has been the subject of a number of prospective randomized studies. These will be presented, along with newer phase II studies. Three randomized studies on the usefulness of HAI after hepatic resection will be presented. PMID- 12075706 TI - The combination of antiangiogenic therapy with other modalities. AB - Angiogenesis is critical for a number of physiologic and pathophysiologic processes, and several angiogenesis inhibitors are now in clinical trials for the treatment of cancer. Antiangiogenic therapy offers a number of potential benefits including lack of drug resistance for some agents, synergistic interaction with other modalities, lack of significant toxicity compared with conventional agents, and a potent antitumor effect. However, no angiogenesis inhibitor has been approved for clinical use. Although antiangiogenic agents offer great therapeutic potential, preclinical and early clinical trial results suggest that these agents will have a delayed onset of activity and may induce stabilization of disease, and not regression, in patients with advanced disease. Studies suggest that regulation of angiogenesis in various capillary beds may be differentially regulated, suggesting that antiangiogenic therapy may require organ-specific optimization. By combining antiangiogenic agents with each other and/or with other modalities used in the treatment of cancer, the limitations of each therapeutic approach will be overcome, leading to enhanced efficacy with diminished toxicity. However, the optimal strategies forthe use, monitoring, and validation of antiangiogenic agents in the clinic remains unclear. Before these agents can be integrated into clinical practice, a better understanding of their mechanism of action and regulation is PMID- 12075707 TI - Infective endocarditis: prevalence, characteristics and mortality in Khon Kaen, 1990-1999. AB - OBJECTIVE: To update the prevalence, characteristics and mortality of infective endocarditis in a tertiary care hospital in Khon Kaen, Thailand. BACKGROUND: Numerous studies have shown that the prevalence and course of infective endocarditis remain unchanged inspite of the advance in treatment, diagnosis and of prophylactic recommendations. METHOD: The authors identified 160 patients from 1990-1999 with 86 per cent definite and 14 per cent possible endocarditis based on the Duke criteria. Data were collected from two tertiary care hospitals. RESULTS: The prevalence was 4 patients per 1,000 hospital admissions. The mean age of the 106 male and 54 female patients was 39+/-16 years. Twenty-four percent of the patients had no previously known heart disease. Native valve endocarditis was present in 95 per cent and prosthetic valve endocarditis was diagnosed in 5 per cent. Infective endocarditis was located on the aortic valve in 42 per cent, the mitral valve in 43 per cent, both mitral and aortic valves in 9 per cent and the tricuspid valve in 8 per cent. The infective organism was identified in only 62 per cent of cases. Streptococci was the most common in 43 per cent, followed by Staphylococci in 16 per cent. During the first month after admission, 45 per cent of the patients underwent surgery. In-hospital mortality was 25 per cent. CONCLUSION: Despite improved diagnostic techniques and aggressive surgical therapy, infective endocarditis remains a serious problem associated with a high mortality in Khon Kaen. PMID- 12075708 TI - Clear topical ointment decreases transepidermal water loss in jaundiced preterm infants receiving phototherapy. AB - The evaporation rate (ER) from the skin was measured in 40 jaundiced preterm infants born at less than or equal to 34 weeks of gestation. The baseline measurements were executed in both the right and left side in 3 positions: upper arm, back and lower leg. The patients were randomly recruited to a treatment or a control group. The treatment group received 3.0 ml of clear topical ointment just before phototherapy. Conventional phototherapy was placed above the incubators in both groups. ER and ambient skin temperature were measured at the same point at 30 minutes and 5 hours during phototherapy. In the control group, ER was increased by 8.0 per cent (P value = 0.01) and 14.5 per cent (P value < 0.001) at 30 minutes and 5 hours during phototherapy, respectively. In the treatment group, clear topical ointment decreased ER by 19.2 per cent (P value < 0.001) and 13.2 per cent (P value = 0.003) at 30 minutes and 5 hours during phototherapy, respectively. Ambient skin temperature during phototherapy was increased significantly (P < 0.01) in both groups. Serum microbilirubin difference of pre and post phototherapy at 24 hours of phototherapy between the 2 groups was not significantly different (P = 0.38). The authors concluded that conventional phototherapy, in premature infants nursed in an incubator, increased transepidermal water loss (TEWL) significantly and the application of clear topical ointment on the skin of jaundiced preterm infants receiving conventional phototherapy in incubators reduce TEWL significantly, without effect on serum microbilirubin. PMID- 12075709 TI - Intraosseous proliferative sparganosis: a case report and review of the literature. AB - Intraosseous proliferative sparganosis is an extremely rare parasitic disease in which the larvae of incomplete differentiated sparganum proliferate in the human bone. We present the first case of intraosseous proliferative sparganosis arising in the long bone. The patient was a 51-year-old man who complained of a slow growing painful mass on his right leg. The radiographic findings showed an infiltrative osteolytic lesion with speckled calcification at the proximal tibia the clinical diagnosis of which favored chondrosarcoma. Incisional biopsy revealed an innumerable number of small globular shapes, whitish parasites. Histologically, the parasites were composed of a few layers of smooth muscle and several calcerous bodies that were enclosed within a single row of tegumental cells. The latter exhibited a wavy appearance and coated with microvilli. These morphologic findings confirmed the nature of these maldifferentiated larvae. The patient was treated by partial resection of the lesion. This should remind clinicians that parasitic infection of the bone can produce a tumor-like lesion. PMID- 12075710 TI - Enuresis in children 5-15 years at Queen Sirikit National Institute of Child Health. AB - A cross-sectional descriptive study was conducted to compare 179 enuretic and 811 non-enuretic patients at Queen Sirikit National Institute of Child Health from July 1998 through December 1998. The objective was to study the epidemiology, risk factors, behavioral and learning problems associated with enuresis in children 5-15 years old. Chi-squared analysis was used. The prevalence of primary, secondary and total enuresis was 15.4, 2.7 and 18.1 per cent respectively in 5-15 year old children. Of these, 88.5 per cent had nocturnal enuresis, 0.6 per cent had diurnal enuresis, 10.6 per cent had diurnal and nocturnal enuresis. Risk factors which significantly correlated with enuresis were the history of bed-wetting in the parents/sibling and inconsistent toilet training. We found that enuresis was not related to sex, birth order, LBW, socioeconomic, marital and parental educational status. Fifty-eight per cent of the parents thought that the enuretic problem needed further treatment. PMID- 12075711 TI - Severe upper gastrointestinal hemorrhage in the newborn. AB - Three infants with severe upper gastrointestinal hemorrhage with esophagogastroduodenoscopic (EGD) findings were reported. The underlying conditions of these infants included Down's syndrome, hypoplastic left heart, and diaphragmatic hernia. The precipitating factors were identified in all cases, including prenatal stress, hypoxemia, prolonged ventilatory support, and gastroesophageal reflux. The EGD findings were composed of multiple gastric ulcers and a duodenal ulcer in the first 2 cases, whereas esophagitis and gastritis were noted in the last case. These ulcers were classified as secondary peptic ulcers. All cases responded well to medical treatment, including ranitidine, sucralfate, omeprazole, cisapride, and octreotide. PMID- 12075712 TI - Myxoma of the nasal cavity and paranasal sinuses: report of a case. AB - Myxomas are benign connective tissue tumors composed of stellate and spindle shaped cells with benign-appearing nuclei lying in a myxoid stroma. They are usually found in the heart, soft tissues, and bones. Myxomas of the nasal cavity and paranasal sinuses are very rare. They are benign but of a locally invasive nature. Recurrent rate is high because of the difficulty to excise them completely. This article presents a case of myxoma of the nasal cavity and paranasal sinuses that extensively invaded the brain and orbits causing blindness. After three resections, also with a transbasal craniotomy in the second operation, the patient remains free from the disease. PMID- 12075714 TI - Fatal myeloencephalitis following yellow fever vaccination in a case with HIV infection. AB - A 53 year old physically healthy man, unaware of any immunocompromised condition developed rapidly fatal myelomeningoencephalitis following a live-attenuated yellow fever vaccination. He was found to have asymptomatic HIV infection with high viral loads and low CD4 counts. This is the first reported case of such an incidence in the world literature. It is strongly suggested that in countries where HIV infection is endemic, an HIV blood test should be performed prior to the yellow fever vaccination and the vaccine should not be given to those immunocompromised persons. PMID- 12075715 TI - Two standards (favoritism). PMID- 12075713 TI - Anesthesia for one-stage bilateral pheochromocytoma resection in a patient with MEN type IIa: attenuation of hypertensive crisis by magnesium sulfate. AB - Multiple endocrine neoplasia (MEN) type IIa, manifesting as an autosomal dominant trait, consists of medullary thyroid carcinoma, parathyroid adenoma or hyperplasia, and pheochromocytoma. We report our experience of a 42-year-old woman, MEN type IIa with a large bilateral pheochromocytoma, who underwent one stage bilateral tumor resection under a combined continuous epidural technique with 0.25 per cent bupivacaine and general anesthesia using vecuronium, fentanyl, nitrous oxide, and isoflurane. An initial intra-operative hypertensive response was acceptably controlled by nitroprusside and a beta-blocker but during tumor handling the hypertensive crisis worsened and she developed acute pulmonary edema despite a continuing high dose of nitroprusside infusion. After receiving intermittent i.v. MgSO4 up to 3 g in 15 min, her condition gradually improved and the cardiovascular response was under control throughout the period of tumor handling. Hypotension encountered post-pheochromocytoma resection was treated by volume replacement, metaraminol, CaCl2, and dopamine infusion. The patient's post operative course was uneventful. PMID- 12075716 TI - B vitamins, vitamin C and hematological measurements in overweight and obese Thais in Bangkok. AB - The dynamic changes of socio-economics leading to the industrialisation of countries are known to affect lifestyle and nutritional behaviours of the population. Review of the literature on the prevalence of obesity showed increasing numbers of the overweight and obese during the past decade. However, information on health and nutritional status of the obese in Thailand has not been widely publicized. This study reveals the vitamin status and hematological picture in 270 overweight and obese Thais in Bangkok, Thailand, compared with 175 normal subjects. No statistically significant differences in haemoglobin and hematocrit were observed in the overweight compared with the control subjects. The prevalence of anaemia was 9.8 per cent among male and 17.2 per cent among female overweight and obese subjects compared with 2.6 per cent and 21.2 per cent in male and female normal controls using the cut-off point of haemoglobin concentration as an indicator of anaemia. Prevalence of hypertension was exhibited in both male and female overweight and obese subjects. Even if there were no statistically significant differences in vitamin B1, B2 and B6 in overweight and obese subjects compared with the controls, high percentages of vitamin C and vitamin B2 deficiencies were observed. Vitamin B2 deficiency was detected in 19.7 per cent of overweight and obese males as well as in 28.7 per cent of overweight and obese females using glutathione reductase activity coefficient (alpha EGR) < 1.5 as the cut-off point. However, clinical signs of vitamin B2 deficiencies were rare. There was also a high percentage of vitamin C (antioxidant vitamin) deficiency in 51.5 per cent of the overweight and obese subjects and 41.7 per cent of the controls, respectively. The results suggest more attention should be paid to health study and nutritional problems for the overweight and obese population, especially concerning vitamins and oxidative stress. Further research is still needed in these aspects. PMID- 12075717 TI - Serum muscle enzymes, muscle pathology and clinical muscle weakness: correlation in Thai patients with polymyositis/dermatomyositis. AB - The clinical correlation between serum muscle enzymes, muscle pathology and muscle weakness was studied in 100 Thai patients (22 males and 78 females) with polymyositis (PM) and dermatomyositis (DM). Their mean +/- SD age and duration of disease were 45.0 +/- 13.9 years and 6.3 + 13.4 months, respectively. There was idiopathic PM in 37 cases, idiopathic DM in 13, PM/DM associated with malignancy in 5 and PM associated with connective tissue disease in 45. Serum muscle enzymes including creatine phosphokinase, lactate dehydrogenase and aspartate aminotransferase were elevated in 87 per cent, 92 per cent, and 82 per cent of cases, respectively. Abnormal electromyographic findings that were compatible with inflammatory myopathy were found in 76 per cent of cases. Seventy-seven per cent had an abnormal muscle biopsy that was consistent with polymyositis. There was a significant correlation between serum muscle enzymes and muscle pathology (p < 0.01). The degree of muscle weakness correlated better with the degree of muscle destruction (p = 0.01) than the degree of muscle inflammation (p = 0.03). The erythrocyte sedimentation rate showed no correlation with serum muscle enzymes, muscle pathology or muscle weakness. PMID- 12075718 TI - The efficacy of a muscle exercise program to improve functional performance of the knee in patients with osteoarthritis. AB - Osteoarthritis (OA) is a common chronic and progressive degenerative joint condition. A major consequence of knee OA is physical disability; especially difficulty with activities requiring ambulation and transfer, which are necessary to maintain independence and a good quality of life. The purposes of this study were to determine the efficacy of a muscle exercise program along with education about knee care, and investigate the clinical factors which influence its therapeutic efficacy. A prospective study was carried out among elderly Thai people aged 60 years and over, living in an urban community of the Bangkok metropolitan area around Siriraj Hospital, between October 1997 and September 1999. The number of elderly people with osteoarthritis of the knee was 392 (male:female=86:306) with a mean age of 67.7+/-6 years. They were interviewed for demographic data and information about their symptoms. The range of the knee movement, quadriceps strength and the six-minute walking distance and a knee X ray were evaluated. Group exercise was arranged twice a week for 8 weeks for the intervention group only. The results of the study revealed that the quadriceps strength in the intervention group had a tendency to increase and the walking distance was significantly improved especially in the first two months but there was a tendency to decline by the sixth and twelfth month after enrollment. On the other hand, there was no statistically significant difference of both important outcomes for the control group when assessed in the second month. The findings at the sixth and twelfth month after enrollment were that the quadriceps strength was slightly increased but the walking distance was decreased when compared with the initial assessment. An exercise frequency of 12 sessions in two months was sufficient to improve muscle strength and walking distance. Group exercise produces a significant improvement in strength and walking ability, especially in the first two months. Deterioration over time could be due to multiple factors, such as lack of regular exercise, lack of motivation, lack of family support or poor economic status, so we should encourage the elderly to exercise regularly. PMID- 12075719 TI - Insulin-like growth factors and their binding proteins in children with IDDM. AB - The structure of insulin-like growth factor (IGF), especially IGF-I, and its receptor is similar to that of insulin. Therefore, the changes of IGFs and IGF binding proteins may be related to glucose homeostasis in children with insulin dependent diabetes mellitus (IDDM). Sixty-three fasting blood samples of 21 children with IDDM attending 3 consecutive diabetic clinics were studied. The HbA1c progressively decreased from the 1st to the 3rd visit. IGF-I levels, both total and free forms, were not significantly different from that of control. IGFBP-3 levels in 3 visits (3406+/-305, 3376+/-252, and 2406+/-247 ng/mL) were significantly lower than that of control (5020+/-415 ng/mL) with the p value of 0.007, 0.002, and < 0.001 respectively. IGFBP-1 levels in the 1st and 2nd visits (102.1+/-12.9 and 114.1+/-14.5 ng/mL) were significantly higher than that of control (60.1+/-15.2 ng/mL) with the p value of 0.03 and 0.01 respectively, but not in the 3rd visit. IGF-I level had a positive correlation with IGFBP-3 (R=0.56, p=0.01) and free IGF-I (R=0.53, p=0.01). Free IGF-I had a negative correlation with IGFBP-1 (R=-0.64, p=0.01). IGF-II at the 15 visit had a negative correlation with HbA1c (R=-0.49, p=0.047). The authors found no correlations between IGF-I, IGFBP-3, free IGF-I, IGFBP-1 and HbA1c in the study. The patients' height SDS followed the genetic height potential. It was, therefore, postulated that a near normal free IGF-I level in diabetic children resulted from a balance of interaction between IGFBP-1 and IGFBP-3 to total IGF-I in order to keep the normal metabolic status as much as possible. PMID- 12075720 TI - Suspended particulate matter in an office and laser smoke particles in an operating room. AB - Suspended particulate matter in an office and laser smoke particles in a laser operative room of the Otolaryngology Department, Ramathibodi Hospital were compared. Suspended particulate matter sizes of PM15, PM10 and PM2.5 were selected due to their impact on health. The amount and sizes of the particles were measured by a laser diode portable dust monitor. The mean and standard deviation were measured every hour for 6 periods and calculated by specific computer software. The amount of suspended particulate matters in the office were within the accepted safety level. The amount of suspended particulate matter including laser smoke particles in the operative room before, during and after each laser evaporative procedure was much higher than that of the office. The amount of suspended particulate matter was dangerous for all personnel in the operative room. Risk management for patients in the operative room should be stressed. The ventilation system of the operative room will be assessed further. PMID- 12075721 TI - Serum estradiol and follicle-stimulating hormone levels in Thai women post total abdominal hysterectomy and bilateral oophorectomy using oral 17 beta-estradiol. AB - OBJECTIVE: To assess the difference of serum estradiol (E2) and follicle stimulating hormone (FSH) levels in Thai women post total abdominal hysterectomy and bilateral oophorectomy, before and after using a daily dose of 2 mg oral 17 beta-estradiol. STUDY DESIGN: Experimental study (before and after treatment). SETTING: Menopause Clinic, Department of Obstetrics and Gynecology, Faculty of Medicine, Chulalongkorn University. SUBJECTS: Thirty-five women who had undergone total abdominal hysterectomy and bilateral oophorectomy at Chulalongkorn Hospital 1 week previously due to benign gynecologic conditions were recruited in the study. Body mass index was 20-25 kg/m2. These women had no contraindication for using hormonal replacement therapy and no history of any hormonal intake in the past. INTERVENTION: All subjects were assigned to receive a daily dose of 2 mg oral 17 beta-estradiol applied at bedtime (8.00 p.m.). MAIN OUTCOME MEASURES: Serum E2 and FSH were measured before and after the study at weeks 4, 8 and 12, 12-14 hours after oral application. The hormonal measurement was performed using the time-resolved fluoroimmunoassay (FIA) method. RESULTS: Five cases were excluded, three cases due to poor compliance which was less than 85 per cent and two cases due to loss to follow-up. Of the remaining 30 cases, the mean age and body mass index were 43.03 +/- 4.58 years and 22.72 + 1.86 kg/m2, respectively. Serum E2 level significantly increased from baseline value at 4, 8 and 12 weeks (median of E2 value at 0, 4, 8 and 12 weeks: 20.00, 22.50, 324.65 and 355.35 pmol/L, p<0.001). On the other hand, there was no significant difference in the FSH serum level (median of FSH value at 0, 4, 8 and 12 weeks: 18.65, 18.40, 18.60 and 20.35 IU/L, p=0.517). CONCLUSION: A daily dose of 2 mg oral 17 beta-estradiol taken at bedtime (8.00 p.m.) for 12 weeks increased the serum E2 to the follicular phase level of the normal menstrual cycle. On the other hand, there was no significant difference in the FSH serum level. PMID- 12075722 TI - Vesicourethral strictures after radical prostatectomy: review of treatment and outcome. AB - OBJECTIVE: Stricture of the vesicourethral anastomosis is a complication after radical prostatectomy. Urethral dilatation, internal urethrotomy, transurethral resection or laser therapy have been reported as the treatment for this complication. The objective of this study was to present our experience with the management of the vesicourethral stricture. MATERIAL AND METHOD: A retrospective study of 90 patients undergoing radical prostatectomy for localized prostatic carcinoma was done. The vesicourethral stricture was treated by dilatation, internal urethrotomy, and transurethral resection of scar tissue in all of the patients. Dilatation was done in less severe cases, internal urethrotomy was done in partial obliteration or after failure of dilatation. Transurethral resection was done in cases with long scar tissue. RESULTS: Ten patients (11%) had anastomotic stricture. The strictures were treated by dilatation in 5 cases, internal urethrotomy in 4 cases and transurethral resection in 1 case. Cure was achieved in all of the patients without incontinence. The median follow-up was 10 months (4-36 months) CONCLUSION: The dilatation and endoscopic procedures of the vesicourethral stricture showed a high cure rate and low incidence of incontinence. PMID- 12075724 TI - Dog rabies in Bangkok. AB - Canine rabies remains a serious public health problem in Thailand. The Queen Saovabha Memorial Institute (QSMI) of the Thai Red Cross Society is the principal rabies diagnostic center in central Thailand. The retrospective study of canine rabies cases collected between 1994-1999 revealed that: (1) The prevalence of rabid dogs has decreased, and was associated with an overall decrease in the number of animals submitted for examination. However, the percentages of dogs with positive Fluorescent antibody test (FA) remained the same at approximately 40 per cent. (2) About 62 per cent of rabid dogs were less than one year old. Dogs at this age are more active and most have not been adequately vaccinated. Approximately 80 per cent of rabid dogs had never been vaccinated against rabies. PMID- 12075723 TI - Antibodies to leptospirosis in rodents from Thailand using a modified human diagnostic assay. AB - The number of reported cases of Leptospirosis in Thailand has grown since 1996. Identification of major reservoirs and endemic areas is essential in surveillance of Leptospira species in Thailand. To assist in the effort of surveillance, a dipstick assay for detecting Leptospira antibodies in mammals was adapted from a human diagnostic assay and tested in a field trial in Thailand. Antibodies to Leptospira were detected in 18 of 60 wild rodents. Four of 9 culture positive rodents were positive by the dipstick assay. The proportion of sera positive for antibodies by dipstick was correlated with positive culture outcome using McNemar test for correlated proportions (0.83, P> 0.05). The dipstick assay was effective in detecting antibodies to Leptospira in mammals and may be useful in resource poor areas or under circumstances where the microagglutination test (MAT) is not practical. PMID- 12075725 TI - Risk of rabies after mammal bites in Thai children. AB - The data were collected by questionnaire at the Outpatient Department of Chulalongkorn University Hospital, Bangkok Thailand. This study revealed that 52 per cent of children had pet mammals in their home of which 67 per cent were dogs. 23.6 per cent of these children gave at least one history of a mammal bite inside (53.4%) or outside (46.6%) their house. Mammal bites of children could be found at all ages. However, most were in the age group of 10-14 years (42.3%), and 5-9 years (39.7%). The most common site of injury was on the legs (56.6%) and hands (30.7%). 31.7 per cent and 68.3 per cent of the bitten children incurred WHO category II and III potential rabies exposures (moderate and severe). 61.9 per cent had performed wound cleansing on each bite injury site and 34 per cent did not. 72 per cent of the children who had mammal bites received no post exposure rabies treatment and 85.7 per cent did not make any effort to capture or observe the animal who had bitten them. Only 10.6 per cent observed the animals for 10 days or more. It was concluded that children are at considerable risk of mammal bites and that they are not receiving optimal care in this canine rabies endemic region and that 50 per cent of human rabies cases in Thailand were in children under 15 years of age. PMID- 12075726 TI - The duration time of urine morphine detection in heroin addicts by radioimmunoassay. AB - Heroin is one of the most seriously abused drugs and its consumption is illegal. Therefore, the detection of the drug in addicts has to be highly accurate and reliable. Morphine detection in urine is the most common method to confirm consumption. Several methods of morphine detection in urine are described such as Latex Agglutination Inhibition (LAI), Thin Layer Chromatography (TLC), Radioimmunoassay (RIA), High Performance Liquid Chromatography (HPLC) and Gas Chromatography and Mass Spectrophotometry (GC/MS). In this study, we performed Radioimmunoassay (RIA) because it is rapid and cheap. Sixty-three male heroin addicts, average age 32 years, with an average time of heroin administration of about 3 years were studied. They used on average Q.635 grams of heroin per day. The time of detection in the urine after the last heroin administration to the first urine testing was about 8.5 hours, over 3-4 days. The amount of morphine in the urine was 17,897.9 ng/ml. Nevertheless, we found that morphine could be detected in the urine every day for seven days. The decreasing rate of daily morphine detection in the urine was 88.4, 74.2, 64.2, 57.1, 63.3 and 44.8 per cent. But there was no significant difference in the route of administration. PMID- 12075727 TI - Tension-free vaginal tape procedure for the treatment of stress urinary incontinence: the first experience in Thailand. AB - OBJECTIVE: Tension-free vaginal tape (TVT) is gaining popularity as an effective treatment for genuine stress urinary incontinence. To better understand this procedure including its results, a retrospective study was carried out to determine surgical technique, effectiveness, safety and early results of this new continence procedure. MATERIAL AND METHOD: From January 1999 to July 2000, twenty female patients with the mean age of 52 years old underwent the TVT procedure. All of them were done by a small incision at mid urethra and a special instrument was used to apply a polypropylene mesh supporting the urethra. Urethral catheter was used as urinary drainage which was removed the next day. Operative time, post operative course, voiding patterns and residual urine were recorded. RESULTS: Mean operative time was 32 minutes (range 15-45 minutes). Up to 10 months, all of the patients were subjectively cured. There was no significant per- and post operative complication including blood loss, wound infection and severe pain. Four patients (20%) had marked residual urine (>100 ml) and needed clean intermittent catheterization for the mean of 0.7 week. CONCLUSION: Although the follow-up period was short, the TVT procedure seems to be a safe and effective method for the treatment of stress urinary incontinence. PMID- 12075728 TI - Comparative study between polyethylene glycol and sodium phosphate solution in elective colorectal surgery. AB - OBJECTIVE: The aim of this study was to compare the pre-operative-post-operative complications and patients' tolerance between sodium phosphate solution (NaP) and polyethylene glycol-electrolyte solution (PEG-ES) for elective colorectal surgery. MATERIAL AND METHOD: All colorectal cancer patients treated between August 2000 and May 2001 (NaP group) who received two doses of 45 ml NaP solution with a glass of water were compared to all colorectal cancer patients between July 1997 and July 2000 (PEG group) who received 3 liters of PEG-ES. Patient tolerance, post-operative septic complications, and serum sodium and potassium before and after bowel preparation were assessed. RESULTS: Twenty five patients in the NaP group and eighty six patients in the PEG group were included in the study. Age, gender and surgical procedure were well matched. Patient tolerance to NaP was superior to PEG-ES (p=0.044). There was a significant increase in serum sodium levels (p=0.022) and a significant decrease in serum potassium levels in NaP group (p=0.018) without any clinical sequelae. 35 per cent of the patients in the PEG group had wound infection versus only 10 per cent of patients in the NaP group (p=0.021). CONCLUSION: Sodium phosphate solution was safe, rapid, well tolerated and could be the standard pre-operative mechanical bowel preparation for elective colorectal surgery. PMID- 12075729 TI - The birth order and personalities of medical students. AB - The purpose of this study was to determine how birth position i.e. first-born, middle-born, lastborn, and only child, correlates with personality. MATERIAL AND METHOD: One hundred and eighty from 186 (97%) 1st year medical students of Chiang Mai Medical School were asked to complete a questionnaire and take a personality test. The data obtained included age, sex, Grade Point Average (GPA), and family background i.e. birth order and the students' personality profiles, which were assessed by the California Psychological Inventory (CPI). RESULTS: The results showed that only children and lastborn had more positive personality factors than other groups. They had more ambition, breadth of interests, versatility, self confidence, clear-thinking, intelligence, and independence than first-born and middle-born (Cs and Ai scale on CPI). In addition, the students whose parents had died, separated, or divorced had some personality profiles that differed from the others. The findings both supported and contradicted other papers. PMID- 12075730 TI - WHO and six publishers launch Access to Research, internet initiative for developing countries. PMID- 12075731 TI - Detection of anthracycline-induced cardiotoxicity: is there light at the end of the tunnel? PMID- 12075732 TI - High-dose chemotherapy for breast cancer: any use for it? PMID- 12075733 TI - The clinical role of somatostatin analogues as antineoplastic agents: much ado about nothing? AB - BACKGROUND: Somatostatin (SST) analogues represent a novel approach for the treatment of certain cancers. The objective of this article is to summarise the current knowledge on SST analogues in the treatment of cancer patients. METHODS: Computerised (Medline) and manual searches were performed to identify publications on clinical trials published in the English-speaking literature between 1966 and 2000. Information abstracted included patients' pre-treatment status, histology, SST receptor (SSTR) evaluation, type of SST analogue, application schedule and dose, duration of treatment, side-effects, response criteria applied (i.e. WHO response criteria, biochemical criteria or symptomatic investigations) and survival. RESULTS: Our search disclosed 22 case reports, five phase 1 and 47 phase II trials, and eight randomised clinical trials using SST analogues (octreotide, lanreotide and vapreotide) as antineoplastic agents. With regard to the phase II trials, conflicting results have been demonstrated in almost all tumour entities investigated. The few randomised studies published so far have shown an impact on survival in patients with hepatocellular cancer, while the effect attributed to treatment in patients with gastrointestinal adenocarcinomas might well have been due to an exceptionally short survival in the control group. There appears to be evidence that SST analogues are able to enhance the therapeutic effects of hormonal intervention in patients with breast cancer, prostate cancer and probably pancreatic cancer. Interpretation of the findings, however, is complicated by the fact that patients were heavily pre treated in some studies and response criteria have not been uniformly applied. In addition, most studies have not been designed to distinguish between receptor mediated (direct) and indirect effects of SST analogues in tumour patients. CONCLUSIONS: According to the results obtained so far, there can be no doubt about the wide therapeutic index and the high efficacy of SST analogues in the symptomatic management of neuroendocrine tumours. Apart from these indications, the data do not justify recommendation of SST analogues as antineoplastic agents outside of clinical trials, as the optimal dose and schedule of application for antineoplastic activity has not been defined for currently used agents. Carefully designed clinical trials including investigation of SSTR status before treatment, evaluation of an indirect mechanism of SST analogues, and assessment of optimal combination of hormone therapy and chemotherapy with SST analogues are clearly needed in the near future. PMID- 12075734 TI - Cyclooxygenase-2 (COX-2), aromatase and breast cancer: a possible role for COX-2 inhibitors in breast cancer chemoprevention. AB - Interest in chemoprevention in oncology using suppressants of prostaglandin (PG) synthesis has been stimulated by epidemiological observations that the use of aspirin and other non-steroidal inflammatory drugs (NSAIDs) is associated with reduced incidence of some cancers, including cancer of the breast. The main target of NSAID activity is the cyclooxygenase (COX) enzyme. Two isoforms of COX have been identified: COX-1, the constitutive isoform; and COX-2. the inducible form of the enzyme. COX-2 can undergo rapid induction in response to many factors such as bacterial lipopolysaccharides, growth factors, cytokines and phorbol esters. COX-2 is overexpressed in some malignancies including carcinoma of the breast. It has been suggested that such enhanced expression may lead to increased angiogenesis such that the inhibition of COX-2 might have a general anticancer effect via decreased blood vessel formation. In addition, an association between COX-2, its main product PGE2 and aromatase activity in human breast cancer suggests that such inhibitors might have an additional, specific prophylactic mechanism for this tumour. New COX-2 inhibitors are already licensed for use in the treatment of arthritis and are well tolerated. Their potential role in chemoprevention of mammary carcinogenesis in rats has already been investigated. What remains to be seen is if these findings can be extrapolated to human studies. PMID- 12075735 TI - Characteristics associated with long-term progression-free survival following high-dose chemotherapy in metastatic breast cancer and influence of chemotherapy dose. AB - BACKGROUND: The purpose of this study was to characterize long-term progression free survivors (LTPFS) of metastatic breast cancer (MBC) following high-dose chemotherapy (HDCT) with autologous stem cell transplantation (ASCT) and to assess the influence of chemotherapy dose in order to identify patients who derive major benefit from this approach. PATIENTS AND METHODS: We compared patient and tumor characteristics of 16 LTPFS with the characteristics of 118 MBC patients who received HDCT with ASCT at our institution between 1992 and 2000. To estimate the cumulative dose of chemotherapy received, the summation dose intensity product (SDIP) of the different chemotherapy regimens was calculated as recently described by Hryniuk et al. The SDIP of the induction regimens was added to that of the HDCT regimens to yield the total SDIP of the chemotherapy received. Multivariate analysis was performed to describe the influence of the total SDIP and other prognostic factors on progression-free survival (PFS). RESULTS: LTPFS were mostly < or = 50 years of age and had limited, chemotherapy sensitive, hormone-responsive MBC. Due to an apparent dose-survival relationship, an increase by 10 units (U) in the SDIP increased the PFS time by 3 months. Independent predictors of an improved PFS were positive estrogen receptors (P = 0.001), positive combined hormone receptors (P = 0.020), and a complete remission/no evidence of disease status after HDCT (P < 0.001). In patients who had a disease-free interval (DFI) >24 months after primary surgery, an SDIP of >55 U was independently associated with a longer PFS [hazard ratio (HR) = 2.73; 95% confidence interval 1.29-5.81; P = 0.009]. CONCLUSION: HDCT can achieve long term PFS in young MBC patients with limited, hormone-responsive and chemotherapy sensitive disease. After a DFI >24 months, a longer PFS is associated with a higher chemotherapy dose as measured by SDIP. These retrospective analyses suggest SDIP might be a tool for studying cumulative dose as a determinant of outcome of MBC chemotherapy. Thus far, however, we cannot clearly identify any subgroup of MBC patients in whom HDCT with ASCT is of particular benefit. PMID- 12075736 TI - Randomized trial of high-dose chemotherapy and hematopoietic progenitor-cell support in operable breast cancer with extensive lymph node involvement: final analysis with 7 years of follow-up. AB - BACKGROUND: The aim of this study was to present an update of overall (OS) and disease-free survival (DFS) and to evaluate the correlation between outcome and pathological findings at surgery in a randomized trial of high-dose chemotherapy following neoadjuvant chemotherapy and surgery in high-risk breast cancer patients. PATIENTS AND METHODS: Ninety-seven women <60 years of age with breast cancer and extensive axillary lymph node involvement received three courses of FE120C (5-fluorouracil 500 mg/m2, epirubicin 120 mg/m2, cyclophosphamide 500 mg/m2) followed by surgery. Eighty-one patients were randomized to receive either a fourth FE120C course alone or a fourth FE120C course followed by high-dose chemotherapy (cyclophosphamide 6 g/m2, thiotepa 480 mg/m2, carboplatin 1600 mg/m2). We performed a univariate analysis on possible prognostic factors and analyzed the sites of relapse. RESULTS: After a median follow-up of 6.9 years, 47 (48%) patients were alive, of whom 36 (38%) were without disease. Sixty patients relapsed after treatment. One patient died of myelodysplastic syndrome, without a relapse. In intention-to-treat analysis, the 5-year DFS rates were 47.5% in the conventional treatment arm and 49% in the high-dose arm, and the 5-year OS rates were 62.5% and 61%, respectively. In the univariate analysis, the clinical T stage before chemotherapy and the number of tumor-positive axillary lymph nodes after induction chemotherapy (P = 0.027) were significant prognostic factors for OS. The same factors (both P = 0.06) plus the estrogen receptor (P = 0.08) were borderline significant factors for DFS. CONCLUSIONS: After a median follow-up of 6.9 years there was no difference in OS or DFS rates between the two treatment groups. The number of tumor-positive axillary lymph nodes after induction chemotherapy and the clinical T-stage before chemotherapy were significant factors for OS. PMID- 12075737 TI - Functional monitoring of anthracycline cardiotoxicity: a prospective, blinded, long-term observational study of outcome in 120 patients. AB - BACKGROUND: With increasing doses the highly tumoricidal anthracycline drugs cause heart damage. Based on empirical drug limitations about 10-15% of patients will develop congestive heart failure (CHF) with a mortality of -50% within 2 years on digitalo-diuretic therapy alone. To avoid CHF there is a consensus recommendation that cardiac function should be monitored in close connection with anthracycline administration. As no prospective studies in a larger series have been performed, these recommendations are based on retrospective data on small numbers of patients. PATIENTS AND METHODS: In a prospective, blinded observational study 120 patients with advanced breast cancer were followed before, during, and a median 3 years after treatment with epirubicin. They had 604 serial radionuclide measurements of left ventricular ejection fraction (LVEF) that were stored without calculations except in patients who developed a well defined CHF. RESULTS: Anthracycline cardiotoxicity was closely correlated with the cumulative dose, with a great variability in individual susceptibility and a dramatic increase with advancing age. With a delayed onset of 3 months or more, epirubicin induced a threatening, slowly progressive deterioration of cardiac function continuing years after treatment. An actuarial estimation of 59% of the patients experienced a 25% relative reduction in LVEF 3 years after 850-1000 mg/m2 of epirubicin and 20% had deteriorated into a CHF. The patients did not spontaneously regain cardiac function whereas continued therapy with a circadian angiotensin-converting enzyme inhibitor for more than 3 months caused a remarkably potent and long-lasting recovery. CONCLUSIONS: Due to the displaced cardiotoxic manifestation, functional monitoring in close connection with anthracycline administration appears to be a poorly effective method while later monitoring is essential. Current monitoring recommendations should therefore be revised. PMID- 12075738 TI - Myocardial injury revealed by plasma troponin I in breast cancer treated with high-dose chemotherapy. AB - BACKGROUND: High-dose chemotherapy (HDC) has been widely utilized in high-risk breast cancer, but it may induce cardiac toxicity. Cardiac dysfunction may become evident weeks or months after HDC and, to date, no early markers of myocardial injury that are able to predict late ventricular impairment are available. We investigated the role of plasma troponin I (TnI) in this setting. PATIENTS AND METHODS: We measured TnI plasma concentration after HDC in 211 high-risk breast cancer women (46 +/- 11 years, mean +/- SD). According to TnI value (< 0.5 or > or = 0.5 ng/ml), patients were allocated into a troponin positive (TnI+; n = 70) and a troponin negative (TnI-; n = 141) group. All patients underwent left ventricular ejection fraction (LVEF, Echo) examination during the following 12 months. RESULTS: LVEF progressively decreased in the TnI+ group but not in the TnI- group. In TnI+ patients a close relationship between the TnI increase, as well as the number of positive TnI assays, and the maximal LVEF decrement, was found (r = -0.92, P < 0.0001 and r = -0.93, P < 0.0001, respectively). CONCLUSIONS: In our population, the elevation of TnI soon after HDC accurately predicts the development of future LVEF depression. In this setting, TnI can be considered a sensitive and reliable marker of myocardial damage with relevant clinical and prognostic implications. PMID- 12075739 TI - Raltitrexed plus oxaliplatin (TOMOX) as first-line chemotherapy for metastatic colorectal cancer. A phase II study of the Italian Group for the Study of Gastrointestinal Tract Carcinomas (GISCAD). AB - BACKGROUND: To evaluate the safety and efficacy of the novel raltitrexed/oxaliplatin combination (TOMOX) as first-line chemotherapy for patients with advanced colorectal cancer. MATERIALS AND METHODS: Previously untreated patients with metastatic colorectal cancer received raltitrexed 3 mg/m2 plus oxaliplatin 100 mg/m2, both intravenously, on day 1 every 3 weeks. Patients were re-evaluated after every third cycle and chemotherapy was continued up to tolerance or disease progression. RESULTS: Fifty-eight patients from 13 Italian Group for the Study of Gastrointestinal Tract Carcinomas (GISCAD) centers were accrued from September 1999 to November 2000. According to the intention-to-treat analysis from 58 patients, the overall response rate was 50% [95% confidence interval (CI) 38% to 62%], with three complete responses and 26 partial responses. The median overall survival (44 patients currently alive) was >9 months and the median time to disease progression was 6.5 months (range 1-15 months). The main hematological toxicity was grade III/IV neutropenia, which occurred in 17% of patients, while anemia and thrombocytopenia were uncommon. Grade III/IV non-hematological toxicities were transient transaminitis (17% of patients); asthenia (16% of patients); neurotoxicity (10% of patients) and diarrhea (7% of patients). No toxic death was observed, one patient with grade IV asthenia after the first cycle refused chemotherapy. CONCLUSIONS: The results of this study suggest that the TOMOX combination is an effective and well tolerated regimen for the treatment of advanced colorectal cancer. Its ease of administration and patient tolerance warrant further investigation as an alternative to fluoropyrimidine-based regimens with repeated and prolonged fluorouracil infusions. PMID- 12075740 TI - Vinorelbine and cisplatin in metastatic squamous cell carcinoma of the oesophagus: response, toxicity, quality of life and survival. AB - BACKGROUND: Vinorelbine and cisplatin are active against squamous cell oesophageal carcinoma. The purpose of this phase II study was to evaluate the efficacy and safety of vinorelbine plus cisplatin in previously untreated patients with metastatic squamous cell oesophageal carcinoma and to estimate the progression-free survival, overall survival and quality of life (QoL) of the patient population. PATIENTS AND METHODS: Seventy-one eligible patients were entered into a study of vinorelbine 25 mg/m2 on days 1 and 8 plus cisplatin 80 mg/m2 on day 1, every 3 weeks. Degree of dysphagia relief was monitored and QoL was measured using the EORTC QLQ-C30. RESULTS: All eligible patients were assessed for response and 24 achieved a confirmed partial response (33.8%; 95% confidence interval 23-46); the median duration of response was 6.8 months, progression-free survival was 3.6 months and median survival of the whole group was 6.8 months. Toxicity was mainly related to neutropenia (grade 3/4 in 41% of patients). At cycle 2, 43% of the patients reported at least a moderate improvement in global health status/QoL and 25% experienced a large improvement. CONCLUSIONS: Vinorelbine plus cisplatin represents a well-tolerated active palliative regimen for patients with advanced squamous cell carcinoma of the oesophagus. This combination may offer a better therapeutic index than cisplatin 5-fluorouracil. PMID- 12075741 TI - Neoadjuvant therapy of rectal carcinoma with UFT-leucovorin plus radiotherapy. AB - BACKGROUND: The object of this phase II study was to assess the impact of preoperative external radiation therapy combined with UFT and leucovorin on tumor response, sphincter preservation and tumor control in patients with rectal carcinoma. PATIENTS AND METHODS: Forty-one patients with resectable extraperitoneal rectal adenocarcinoma received radiation therapy and two courses of chemotherapy. Chemotherapy consisted of a 2-h infusion of 6S-steroisomer of leucovorin (6SLV) 250 mg/m2 on day 1, oral 6SLV 7.5 mg every 12 h on days 2-14, and UFT either 350 or 300 mg/m2 on days 1 to 14 every 28 days. Six additional courses of chemotherapy were given after surgery. RESULTS: Seven of 16 patients (43%) who received 350 mg/m2/day of UFT had grade 3-4 diarrhea and two other patients (12%) had grade 3-4 dermatitis. The next 25 patients received 300 mg/m2/day of UFT and only 14% of them had grade 3-4 diarrhea. Surgery consisted of low-anterior resection in 26 patients (63%) and abdominal-perineal amputation in 15 (37%). There were six histological complete responses (15%). Downstaging occurred in 25 patients (63%). The overall survival at 3 years was 90% and the pelvic disease-free survival 92%. CONCLUSIONS: Preoperative therapy with radiotherapy and UFT-6SLV downstaged 63% of tumors and allowed a sphincter preserving procedure in some patients. Toxicity was moderate. This scheme is convenient because of the oral administration of chemotherapy. PMID- 12075742 TI - Phase II trial of pemetrexed disodium (ALIMTA, LY231514) in chemotherapy-naive patients with advanced non-small-cell lung cancer. AB - BACKGROUND: To evaluate the efficacy and safety of pemetrexed therapy for chemotherapy-naive patients with surgically incurable non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Eligible patients received pemetrexed 600 mg/m2 every 3 weeks. Restaging was performed after every two cycles of therapy and toxicity was assessed at each cycle of pemetrexed. In the absence of disease progression or undue toxicity, treatment was continued for a maximum of 12 cycles. RESULTS: Fifty-nine patients (median age 59 years; range 39-74 years) received a median of four cycles of pemetrexed. Nineteen patients (32%) had a ECOG performance status (PS) of two and 39 patients (66%) had stage IV disease. The most common histological sub-types were adenocarcinoma (20 patients, 34%) and large cell (18 patients, 31%). Sixteen patients (27%) had received prior radiotherapy. Nine patients achieved a partial response for an overall response rate of 15.8% (95% confidence interval CI 7% to 28%). The median duration of response was 4.9 months, and the median survival was 7.2 months. The principal toxicities were myelosuppression and rash. While grade 3 or 4 neutropenia was seen in 25 patients (42%), only two patients (3%) developed grade 3 infection. Eighteen patients (31%) developed grade 3 or 4 cutaneous toxicity, which improved with prophylactic oral dexamethasone administered for 3 days beginning the day before pemetrexed treatment. Asymptomatic elevations in hepatic biochemistry (especially alanine transaminase and aspartate transaminase) were seen in 47 patients (80%); however, these did not interfere with the dose or schedule of pemetrexed and returned to normal levels throughout the study. CONCLUSIONS: This is the largest study confirming the encouraging single-agent activity of pemetrexed in chemotherapy-naive patients with NSCLC. In addition, this study demonstrates that a dose of 600 mg/m2 can be delivered safely; however, treatment should be restricted to patients with a PS of 0 or 1. The results of combination studies are awaited with interest. PMID- 12075743 TI - Phase II randomized multicenter study evaluating a treatment regimen alternating docetaxel and cisplatin-vinorelbine with a cisplatin-vinorelbine control group in patients with stage IV non-small-cell lung cancer: GFPC 97.01 study. AB - BACKGROUND: The potential absence of cross-resistance between cisplatin and docetaxel in non-small-cell lung cancer (NSCLC) suggests that alternating regimens of cisplatin-based chemotherapy and docetaxel might increase the activity of chemotherapy in stage IV NSCLC. PATIENTS AND METHODS: Randomized, multicenter, non-comparative phase II study in patients with stage IV NSCLC (Eastern Cooperative Oncology Group performance status of 0-2). Patients randomized to alternating treatment group (A) received docetaxel 100 mg/m2 on days (D) 1 and 43 alternating with cisplatin 100 mg/m2 on D22 and vinorelbine 30 mg/m2 on D22, D29 and D36. Those randomized to the control group (B) received cisplatin 80 mg/m2 on D1, D22 and D43 and vinorelbine 30 mg/m2 once a week from D1 to D57. Treatment was continued for a further 6 weeks in the event of objective response or stabilization. RESULTS: Seventy patients were enrolled (group A: 38, group B: 32). More premature treatment discontinuations due to toxicity were observed in group A (median number of cycles: 3) than in group B (median number of cycles: 5). The intention-to-treat objective response rate was 10.8% [95% confidence interval (CI) 0.8% to 20.8%] in group A compared with 25% (95% CI 10% to 40%) in group B, the median time to treatment failure being 10.2 weeks and 17.3 weeks, respectively. The median survival and 1-year survival were 29.1 weeks and 39% in group A compared with 41.6 weeks and 42% in group B. Febrile neutropenia occurred in 5.9 and 4.9% of the cycles in group A and group B, respectively. Non-hematological toxicity was moderate in the two groups. CONCLUSIONS: The addition of docetaxel alternating with cisplatin-vinorelbine did not enhance the activity of this combination. The development of sequential regimens might be a more promising way of exploiting the absence of cross resistance between these two drugs. PMID- 12075744 TI - Patterns of failure after prophylactic cranial irradiation in small-cell lung cancer: analysis of 505 randomized patients. AB - BACKGROUND: Prophylactic cranial irradiation (PCI) has a beneficial effect on overall survival in patients with small-cell lung cancer (SCLC) in complete remission as shown in a worldwide meta-analysis. The current analysis aimed to evaluate PCI effects on patterns of failure in this patient category. PATIENTS AND METHODS: The Institut Gustave-Roussy coordinated two parallel randomized studies including a total of 511 patients with SCLC. Patients were randomly assigned to either PCI (24 Gy in eight fractions and 12 days) or no PCI. Patterns of failure were analyzed according to (i) total event rates and (ii) isolated first site of relapse using a competing risk approach. RESULTS: Five hundred and five patients were analyzed. The 5-year cumulative rate of brain metastasis as an isolated first site of relapse was 37% in the control group and 20% in the PCI group (P < 0.001). The overall 5-year rates of brain metastasis were 59% and 43%, respectively [relative risk (RR) 0.50; P < 0.001]. The 5-year overall survival rates were 15% in the control group and 18% in the PCI group (RR 0.84; P = 0.06). CONCLUSIONS: PCI decreased significantly the risk of brain metastasis. Other events were not influenced. The relative death risk reduction was of borderline significance. Results reported as isolated first cause of failure and subsequent competing events may explain why a major treatment effect on brain metastases rate has a rather moderate effect on survival. PMID- 12075745 TI - A phase I study of oral uracil-ftorafur plus folinic acid in combination with weekly paclitaxel in patients with solid tumors. AB - INTRODUCTION: Ftorafur is an orally available prodrug of 5-fluorouracil (5-FU). Its combination with uracil in a molar ratio of 1:4 (UFT) increases the 5-FU concentration in tumor cells compared with ftorafur alone. Paclitaxel has a broad spectrum of activity against solid tumors and synergic effects with UFT have been demonstrated in vitro. A phase I study was performed to determine the maximum tolerated dose of the combination of UFT and paclitaxel in patients with advanced solid tumors. STUDY DESIGN: UFT and folinic acid were applied at 300 mg/m2/day and 90 mg/day, respectively, on days 1-28, repeated on day 36. Paclitaxel was applied on days 1, 8, 15 and 22 of each cycle. The starting dose of paclitaxel was 50 mg/m2 and escalation in 10 mg/m2 steps was performed up to 100 mg/m2 weekly. RESULTS: Forty-seven consecutive patients with various solid tumors have been included in six different dose levels. One hundred and thirty cycles have been applied. The treatment was well tolerated overall. Most frequently encountered adverse effects were gastrointestinal and hematological toxicity (diarrhea CTC 3/4 in 6% of patients, anemia in 11%, leukocytopenia in 9%, polyneuropathy in 9%, fatigue in 11%, other in 6%). Partial remissions were observed in 28% of patients. CONCLUSION: Owing to the lack of overlapping toxicities, UFT/folinic acid plus paclitaxel can be combined at doses of proven single agent activity. Side effects are mainly attributable to the gastrointestinal toxicity of UFT and to the neuro- and hematotoxicity of paclitaxel. The recommended doses for phase II studies are 300 mg/m2 of UFT plus 90 mg of folinic acid on days 1-28, and 90 mg/m2 of paclitaxel weekly. PMID- 12075746 TI - Pharmacokinetic study of cystemustine, administered on a weekly schedule in cancer patients. AB - BACKGROUND: Cystemustine is a chloroethylnitrosourea mostly active in humans against glioma and melanoma. The present report describes the results of a new phase I trial with cystemustine administered on a weekly schedule. The pharmacokinetic and pharmacodynamic properties of cystemustine were investigated. PATIENTS AND METHODS: Forty-three patients entered this study. Cystemustine was administered at dose levels ranging from 30 to 60 mg/m2. The drug was given on days 1, 8, 15 and 22, followed by a 4-week rest period. RESULTS: Thrombocytopenia was the dose-limiting toxicity and appeared to be reversible, but probably cumulative. This toxicity appeared dose-related, both in frequency and severity. The maximum tolerated dose was 60 mg/m2. Nonhematological toxicity was generally mild. Three partial responses were observed at dose levels of 50 and 60 mg/m2. Pharmacokinetics analysis showed mono- or biphasic cystemustine blood disposition with a mean a half-life of 4 min and mean terminal half-life of 49 min. CONCLUSIONS: There was a clear linear relationship between the area under the blood drug concentration-time curve (AUC) and the dose of cystemustine (P < 0.001). There was also a significant relationship between the AUC and the toxic effects of cystemustine on platelets, granulocytes and leukocytes (P < 0.001). A reasonable starting dose for phase II studies is 40 mg/m2, with dose escalation based on blood cell counts. PMID- 12075747 TI - Treatment with the novel anti-angiogenic agent PI-88 is associated with immune mediated thrombocytopenia. AB - BACKGROUND: The novel molecule PI-88 is a highly sulfonated oligosaccharide which inhibits heparanase activity and competes with heparan sulfate binding of growth factors such as FGF and VEGF. Preclinical data demonstrates that PI-88 inhibits angiogenesis and has anti-metastatic effects. The aim of this phase I study was to determine the recommended dose and toxicity profile of PI-88. PATIENTS AND METHODS: PI-88 was given intravenously in increasing duration of administration (0.57 mg/kg for 2 h, 0.57 mg/kg/day for 1 day, 4, 7 and 14 consecutive days) and then increasing dose for 14 consecutive days (1.14 mg/kg/day and 2.28 mg/kg/day) in patients with advanced malignancies until dose-limiting toxicity (DLT) was observed. Fourteen assessable patients with advanced malignancies received PI-88 intravenously. RESULTS: DLT was thrombocytopenia. The thrombocytopenia appeared to be immunologically mediated with the development of anti-heparin platelet factor 4 complex antibodies. There were no other significant toxicities. At the final dose and schedule (2.28 mg/kg/day for 14 days), there was limited evidence of biological activity as measured by the surrogate marker activated partial thromboplastin time (APTT), although two patients had stabilisation of disease. CONCLUSIONS: In conclusion, PI-88 at a dose of 2.28 mg/kg/day for 14 days resulted in dose-limiting thrombocytopenia which appeared to be immune related. Limited evidence of biological activity was noted. Alternate scheduling and routes of administration are now being explored. PMID- 12075748 TI - Phase II study of XR5000 (DACA) administered as a 120-h infusion in patients with recurrent glioblastoma multiforme. AB - BACKGROUND: XR5000 is a tricyclic carboxamide that intercalates DNA and inhibits both topoisomerase I and II. The aim of this study was to evaluate the efficacy and tolerability of XR5000 in patients with recurrent glioblastoma multiforme previously untreated with chemotherapy at relapse. PATIENTS AND METHODS: Patients received XR5000 at a dose of 3010 mg/m2 as a 120-h central venous infusion every 3 weeks. An independent panel assessed response every two cycles using McDonald's criteria (tumour size, steroid intake and neurological status); toxicity was graded according to the National Cancer Institute-Common Toxicity Criteria, version 2.0. RESULTS: Sixteen patients were enrolled (one ineligible patient was excluded from efficacy evaluation). Performance status was zero (five patients), one (nine patients) or two (one patient). They received 30 cycles of XR5000 (median 2, range 1-5). Haematological toxicity was mild, with only one patient experiencing grade 3 neutropenia. Other related grade 3/4 adverse events included chest pain (one patient), axillary vein thrombosis (one patient) and rigors/fever in the absence of neutropenia (one patient). There were no objective responses, 14 patients progressing on XR5000 and one having stable disease. CONCLUSIONS: Although XR5000 was generally well tolerated, these results do not support further evaluation in patients with glioblastoma multiforme using this dose and schedule. PMID- 12075749 TI - Quantitative measurement of BCR/abl transcripts using real-time polymerase chain reaction. AB - BACKGROUND: Quantitative real-time polymerase chain reaction (Q-Rt-PCR) is a new tool in the detection and quantification of the BCR/abl fusion transcripts in chronic myelogenous leukemia (CML). This study investigates its specificity, sensitivity and potential clinical usefulness. PATIENTS AND METHODS: Parallel analysis of Q-Rt-PCR and the conventional reverse transcription-mediated PCR (RT PCR) were performed on 567 samples from 481 patients. Treatment response was monitored by Q-Rt-PCR at 6 and 12 months of 61 patients on STI-571 and 103 patients on interferon. RESULTS: The concordance rate between Q-Rt-PCR and RT-PCR was 96.3% (546/567), with 341 positives and 205 negatives. The positive equivalents ranged from 2 x 10(-6) to 1.2 microg of K562 cell RNA. Karyotyping in 372 samples revealed excellent correlation with Q-Rt-PCR measurements (P < 0.001). Setting residual BCR/abl < 0.01 as an early goal of molecular response, we observed that STI-571 induced a better response than interferon: 49% (20 of 41 patients) versus 35% (15 of 62 patients) at 6 months (P = 0.025) and 52% (32 of 61 patients) versus 34% (35 of 103 patients) at 12 months (P = 0.01), respectively. CONCLUSIONS: Q-Rt-PCR provides reliable measurements of BCR/abl fusion transcripts. It is potentially useful in assessing molecular residual disease after therapy. PMID- 12075750 TI - Urate-oxidase in the prevention and treatment of metabolic complications in patients with B-cell lymphoma and leukemia, treated in the Societe Francaise d'Oncologie Pediatrique LMB89 protocol. AB - PURPOSE: To evaluate the frequency of metabolic complications and dialysis due to tumor lysis syndrome in patients with B-cell advanced-stage non-Hodgkin's lymphoma (NHL) and L3 leukemia at initiation of chemotherapy including the use of urate-oxidase. PATIENTS AND METHODS: Retrospective review of the clinical records of 410 patients with stage III and IV B-cell NHL and L3 leukemia treated in France and prospectively registered in the LMB89 protocol. RESULTS: During the first week of chemotherapy, only 34 of 410 patients recorded metabolic problems that included hypocalcemia (< 70 mg/dl) in 24 patients, hyperphosphatemia (> 6.5 mg/dl) in 28 and elevation of creatinine > or = 2 SD in 16. Six patients underwent dialysis for life-threatening problems and a seventh as a preventive measure. In the other 27 cases, metabolic problems were successfully resolved using urate-oxidase in combination with alkaline hyperhydration. Among the 410 patients, one case of hemolysis was reported and there was no severe allergic reaction to urate-oxidase. CONCLUSIONS: Only 1.7% of patients in our study receiving urate-oxidase during their induction chemotherapy needed renal dialysis. Urate-oxidase was well tolerated, and used as prophylaxis and/or treatment of hyperuricemia and tumor lysis syndrome consistently gave a lower rate of renal and metabolic complications than in other series of similar patients. PMID- 12075751 TI - Capecitabine can induce acute coronary syndrome similar to 5-fluorouracil. AB - Capecitabine is a member of a new class of oral fluoropyrimidines. It is a 5 fluorouracil (5-FU) prodrug, activated by a series of enzymes. Activation has been demonstrated to occur preferentially in tumor tissue, which may explain the favorable balance of efficacy and toxicity of this drug. Cardiotoxicity, a rare but potentially serious adverse effect of 5-FU, has not been reported for capecitabine to date. Here we report a patient who experienced a severe and prolonged acute coronary syndrome during treatment with capecitabine. He had previously developed similar symptoms during treatment with infusional 5-FU. Capecitabine should thus be considered an agent with cardiotoxic potential. This adverse effect should be specifically monitored in all patients treated with capecitabine. Patients with symptoms suggestive of cardiotoxicity during previous treatment with a fluoropyrimidine should not be treated with capecitabine. PMID- 12075752 TI - Fusion PET-CT imaging of neurolymphomatosis. AB - In a patient suffering from peripheral neuropathy due to neurolymphomatosis, fused PET-CT imaging, performed on a novel in-line PET-CT system, showed multiple small nodular lesions extending along the peripheral nerves corresponding to an early relapse of a transformed B-cell non-Hodgkin's lymphoma. PMID- 12075753 TI - Chemotherapy with gemcitabine in patients with gall-bladder carcinoma. PMID- 12075754 TI - Highly aggressive leiomyosarcoma associated with Lynch II syndrome: increasing the range of extracolonic cancers related with hereditary non-polyposis colonic cancer. PMID- 12075755 TI - Relative frequency of Haemophilus influenzae type b pneumonia in Chinese children as evidenced by serology. AB - OBJECTIVES: It is commonly held that Haemophilus influenzae pneumonia among children in Asia is mostly caused by serotypes other than b (Hib). If so, Hib conjugate vaccines would play little role in the prevention of pneumonia. In two prospective series of children hospitalized for pneumonia in China, the causative agents were searched for with a wide panel of microbiologic assays. METHODS: In the university hospitals of Beijing and Hefei, 156 consecutive children 3 months of age and older with symptoms and signs of pneumonia were studied. Blood culture, chest radiograph, nasopharyngeal aspirate for viral antigen detection and paired sera for 20 microbiologic assays were taken. Severity was graded, and the course of illness was monitored uniformly. RESULTS: In Beijing only likely contaminants grew from blood cultures, and in Hefei pathogens were identified in two cases. In combined series evidence for bacterial, mixed and viral etiology was obtained in 30, 7 and 21% of cases, respectively. The dominant bacteria were pneumococcus, Hib, Mycoplasma pneumoniae and Chlamydia pneumoniae, responsible for 13, 10, 8 and 8% of cases, respectively. Most patients were treated with extended spectrum antimicrobials such as piperacillin, cefotaxime or ceftriaxone, alone or in combination. One child died. CONCLUSIONS: As in most other series from other countries, the leading agent causing childhood pneumonia was pneumococcus but, in line with our previous experience from Beijing, the second most common agent detected was Hib. This observation suggests great potential for pneumococcal and Hib vaccinations in China. Because no evidence supported the need for routine use of extended spectrum antimicrobials, narrower spectrum agents would be safer for patients, would be cheaper for the community and would offer a way to address increasing resistance problems. PMID- 12075756 TI - Invasive pneumococcal infections in children with asplenia. AB - BACKGROUND: Asplenia is associated with an increased risk of infections caused by Streptococcus pneumoniae. Overwhelming infection can be fulminate and lead to a fatal outcome. OBJECTIVE: To review the epidemiology and clinical course of invasive S. pneumoniae infections in children with asplenia before the release of the conjugate pneumococcal vaccine. METHODS: Children with S. pneumoniae infections from eight children's hospitals in the US were identified prospectively from September, 1993, to August, 1999. Further demographic, medical and microbiologic information was gathered retrospectively from the charts of patients with asplenia. RESULTS: Twenty-two asplenic patients with 26 episodes of invasive S. pneumoniae were identified. This represents 1% of the 2,581 episodes of invasive S. pneumoniae infections identified in our study. Twelve had congenital asplenia (CA), and 10 had undergone surgical splenectomy. Nine of the patients with CA had associated complex congenital heart disease. The median age at first infection was 12.5 months for CA patients as compared with 69 months in children with surgical splenectomy (P < 0.001). Seventy-five percent of those eligible had received the polysaccharide pneumococcal vaccine. The most common serotypes isolated were 6B (8), 23F (7), 18C (2) and 19A (2). Antimicrobial prophylaxis had been prescribed for 82% of the study cohort. Clinical presentations of the 26 episodes included fever (22), shock (7), petechiae or purpura (7), disseminated intravascular coagulation (5) and respiratory distress (5). Clinical illness included bacteremia alone (12), meningitis alone (8), bacteremia with otitis media-sinusitis (3), bacteremia with pneumonia (2) and meningitis with osteomyelitis (1). Five of the 6 patients who died had meningitis. Three of the survivors (19%) had significant morbidity, and all of them had meningitis. Two patients had 2 episodes each, and 1 patient had 3 episodes. All but 1 of the multiple episodes was with a different serotype. Forty six percent of isolates were nonsusceptible to penicillin, and 19% were nonsusceptible to ceftriaxone. There was no association between antimicrobial resistance and mortality. CONCLUSIONS: Invasive pneumococcal disease in patients with asplenia has a high mortality, especially in those with meningitis. Even though the new conjugate vaccine might increase protection, 19% of patients had disease caused by serotypes not included in the current heptavalent vaccine. Clinicians should continue to be aggressive in evaluating asplenic patients with unexplained fevers. PMID- 12075757 TI - Impact of rapid polymerase chain reaction results on management of pediatric patients with enteroviral meningitis. AB - BACKGROUND: Enterovirus (EV) infections can be rapidly detected by PCR. However, several studies suggest that results must be available early in the management of the patient to impact significantly on patient care. We evaluated this hypothesis directly during an outbreak of EV aseptic meningitis. METHODS: From June through November, 1998, EV PCR was performed 5 days a week on cerebrospinal fluid specimens from pediatric patients evaluated for meningitis. We compared antibiotic use, length of stay and hospital charges in a group of patients with EV meningitis whose positive EV PCR results were available within 24 h of specimen collection, to a group of similar patients whose results were available >24 h after collection. RESULTS: Cerebrospinal fluid specimens were submitted for EV PCR from 113 patients with suspected EV meningitis, and 50 of 113 (44%) were positive. Of these 50 EV-PCR-positive patients, 17 of 50 (34%) had EV PCR results available in < or = 24 h and 33 of 50 (66%) had results available in >24 h. Patients with EV-positive results reported < or = 24 h after specimen collection had 20 h less of antibiotic use (P = 0.006) and $2,798 less in hospital charges (P = 0.001) than patients with positive results available in >24 h. Hospitalized patients who received positive results rapidly did not have significantly less antibiotic therapy or shorter length of stay, but hospital charges were reduced by $2,331 (P = 0.009). CONCLUSION: Rapid reporting of PCR results can have a significant impact on several outcome measures for patients with EV meningitis. PMID- 12075759 TI - Lower respiratory tract infections associated with influenza A and B viruses in an area with a high prevalence of pediatric human immunodeficiency type 1 infection. AB - BACKGROUND: Despite the high burden of pediatric HIV-1 infection in developing countries, there are few data on the clinical course of influenza virus associated lower respiratory tract infection (LRTI) in these children. OBJECTIVE: To define and compare the clinical course of HIV-1-infected and -uninfected African children hospitalized with influenza virus associated severe LRTI. METHODS: Children with severe LRTI were prospectively recruited between March, 1997, and March, 1999, as part of a broader study evaluating the etiology and outcome of this condition in hospitalized HIV-1-infected and -uninfected children. The results of children in whom influenza A or B virus was identified by immunofluorescent antibody staining after shell vial culture are reported. Viruses isolated were typed by hemagglutination inhibition assays. RESULTS: Twenty-five (21.6%) of the 116 children hospitalized with severe LRTI in whom influenza A or B virus was identified were HIV-1-infected. HIV-1-infected children were older than uninfected children (mean age +/- SD 17.4 +/- 10.8 months vs. 10.2 +/- 8.9 months; P = 0.002). HIV-1-infected children were more likely to have an underlying medical illness (in addition to HIV-1 infection) predisposing them to more severe LRTI (32.0% vs. 13.2%; P = 0.03). HIV-infected children were also more likely to have indirect evidence of bacterial coinfection, including chest radiographic evidence of confluent alveolar consolidation (78.9% vs. 35.1%, P = 0.006), and were less likely be wheezing (8.0% vs. 31.9%, P = 0.01). However, there was no difference in the clinical outcome of HIV-1-infected and -uninfected children. The duration of hospitalization [median (range) 5 (2 to 33) days vs. 4 (0 to 21) days, P = 0.08] and the mortality rates (8.0% vs. 2.2%, P = 0.20) were similar between HWV-1 infected and -uninfected children. CONCLUSION: HIV-1-infected children hospitalized with severe LRTI associated with influenza virus have an outcome similar to that of HIV-1-uninfected children even in the absence of antiretroviral or anti-influenza virus treatment. PMID- 12075758 TI - Respiratory syncytial virus prophylaxis: cost-effective analysis in Argentina. AB - INTRODUCTION: Respiratory syncytial virus is a major cause of bronchiolitis during the first year of life. Preterm infants and patients with bronchopulmonary dysplasia (BPD) have a higher risk of hospitalization. The objective of this study was to assess the risk of hospitalization and to evaluate the cost effectiveness of palivizumab use in our at risk population. METHODS: The study design was based on the hypothetical use of palivizumab in a cohort of patients. The cost effectiveness analysis included costs of palivizumab, the drug prescription and hospitalization. The effectiveness measure was the hospitalization rate decrease according to the result of the IMpact study: a 55% reduction in the need for hospitalization attributable to RSV. The years 1998 and 1999 were analyzed. Evaluable patients had follow-up at the High Risk Clinic and fulfilled the American Academy of Pediatrics criteria for palivizumab prophylaxis. RESULTS: Forty-two patients, 24 with BPD and 18 preterm infants without BPD, were evaluated. Ten patients required hospitalization because of respiratory syncytial virus; one of them died. Hospitalization cost was $184,777. The total palivizumab administration cost would have been $185,064. With a 55% decrease of hospitalization rate, the cost per hospitalization averted would have been $15,358, and the number needed to treat would have been 7.9. A sensitivity analysis showed that the cost per hospitalization averted could range between $2,171 and $48,630 according to the hospitalization rate. CONCLUSIONS: Because new effective but expensive treatments like palivizumab are available, this cost effective analysis can be an important tool in decisions about resource distribution. PMID- 12075760 TI - Comparison of two dosages of azithromycin for three days versus penicillin V for ten days in acute group A streptococcal tonsillopharyngitis. AB - BACKGROUND: Three-day, 10 mg/kg/day azithromycin (AZM) studies in pediatric acute group A streptococcal tonsillopharyngitis have shown contradictory bacteriologic results. This study investigates the efficacy and tolerability of two dosages of 3-day azithromycin (20 mg/kg/day and 10 mg/kg/day) compared with 10-day penicillin V. METHODS: This was a prospective, comparative, randomized, multicenter trial. Children were scheduled to return for visits at 14 days (main end point) and 1 month after the onset of treatment for clinical and bacteriologic assessment. Molecular tools were used to compare pre- and posttreatment group A beta-hemolytic Streptococcus (GABHS) isolates. RESULTS: Between November, 1997, and July, 1998, 501 patients (169 AZM 10 mg, 165 AZM 20 mg, 167 penicillin V) between 2 and 12 years old were enrolled; 500 were assessable for safety, 469 for intent to treat analysis and 420 for efficacy in the per protocol analysis. Before treatment 25 (7.9%) of 315 GABHS stains isolated from patients receiving AZM were resistant to this compound. On Day 14 pretreatment GABHS were eradicated from 78 (57.8%) of the 135 children receiving the AZM 10 mg regimen, 131 (94.2%) of the 139 receiving AZM 20 mg and 123 (84.2%) of the 146 taking penicillin. One month after the outset of treatment, bacteriologic relapses were observed in 40.5% (n = 30) of the children receiving AZM 10 mg, 14.8% (n = 18) of children taking AZM 20 mg and 13.2% (n = 15) of those treated with penicillin V. AZM 20 mg/kg/day was statistically superior to AZM 10 mg/kg/day microbiologically on Day 14 (P = 0.0001) and Day 30 (P = 0.0001) and clinically on Day 14 (P = 0.0035). AZM 20 mg/kg/day was statistically equivalent both microbiologically and clinically to standard therapy with penicillin V at all endpoints. The incidence of treatment-related adverse events was similar in the two azithromycin groups [AZM 10 mg, 31 of 169 (18.3%); AZM 20 mg, 37 of 164 (23%)] but significantly higher than those observed in the penicillin V group [5 of 166 (3%); P < 0.0001]. Most treatment-related adverse events were gastrointestinal and of mild-to-moderate severity. Fourteen patients withdrew from the trial because of adverse events (1 in the penicillin V group, 7 in the AZM 10 mg group and 6 in the AZM 20 mg group). CONCLUSION: This is the first study to demonstrate a daily dose-dependent difference in microbiologic efficacy of a regimen; 3-day AZM 20 mg/kg/day is a more effective regimen than 3 day AZM 10 mg/kg/day for pediatric GABHS tonsillopharyngitis. PMID- 12075761 TI - Azithromycin versus penicillin V for treatment of acute group A streptococcal pharyngitis. AB - OBJECTIVE: To compare a 3-day azithromycin vs. a 10-day penicillin V regimen for treatment of acute group A streptococcal (GAS) pharyngitis in children and to determine whether viral infection and/or pharyngeal GAS carriage in patients and adult contacts affect clinical and bacteriologic efficacy. METHODS: This multicenter, randomized, comparative, open label study compared 3-day, once daily 10 mg/kg azithromycin oral suspension with a 10-day regimen of 100,000 IU/kg/day penicillin V oral suspension in three divided doses in children with acute GAS pharyngitis. Clinical and bacteriologic efficacy and tolerability of the antibiotics were evaluated. Recurrence of symptoms and infection was monitored for 6 months. RESULTS: In total, 292 children (age range, 2 to 12 years) received at least one dose of study medication. Clinical success (cure/improvement) with either antibiotic was similar at the end of therapy (Day 14; azithromycin, 95%; penicillin V, 97%) and at Day 28 (azithromycin, 94%; penicillin V, 95%). Bacteriologic eradication was significantly less with azithromycin than with penicillin V at Day 14 (azithromycin, 38%; penicillin V, 81%; P < 0.001) and at Day 28 (azithromycin, 31%; penicillin V, 68%; P < 0.001). There was no associated increase in GAS-related sequelae. The lower incidence of bacteriologic eradication with azithromycin was not the result of possible concomitant viral infections in the patients, GAS carriage in one parent/guardian or any reduced susceptibility in pretreatment GAS isolates. Both antibiotics were equally well tolerated. CONCLUSIONS: Treatment with 3-day, once daily 10 mg/kg azithromycin for GAS pharyngitis is associated with similar high levels of clinical efficacy, but lower levels of bacteriologic eradication, than with 10-day 100,000 IU/kg/day penicillin V. PMID- 12075762 TI - Antibody response to an eleven valent diphtheria- and tetanus-conjugated pneumococcal conjugate vaccine in Filipino infants. AB - BACKGROUND: Pneumococcal conjugate vaccines are intended to provide effective protection against pneumococcal infections, but very little information on antibody responses in infants living in countries with high pneumococcal disease burden exists. METHODS: In this study 50 healthy Filipino infants were enrolled at a village health center in Cabuyao to receive 11-valent diphtheria- and tetanus-conjugated pneumococcal vaccine at 6, 10 and 14 weeks of age (primary series) simultaneously with diphtheria-tetanus-whole cell pertussis/polyribosylribitol phosphate conjugated to tetanus toxoid, hepatitis B virus and oral poliovirus vaccines and at 9 months of age (booster dose) simultaneously with measles vaccine. The alum-adjuvanted study vaccine contained pneumococcal polysaccharide of serotypes 1, 4, 5, 7F, 9V, 19F and 23F conjugated to tetanus protein and pneumococcal polysaccharide of serotypes 3, 6B, 14 and 18C conjugated to diphtheria toxoid. Serum samples for enzyme immunoassay analyses were collected at 6, 10 and 14 weeks and 9 and 10 months of age. RESULTS: Very high geometric mean antibody concentrations (GMCs) against most pneumococcal serotypes were observed after the first three doses of vaccine (range, serotype 23F, 3.89 microg/ml to serotype 4, 23.41 microg/ml) with the exception of serotype 6B and 14, with GMCs of 1.12 and 2.18 microg/ml, respectively. The fourth dose increased the GMCs against most serotypes (range, serotype 14, 1.65 to serotype 19F, 33.43 microg/ml). The maternally derived antibodies did not decrease the response to the vaccine. CONCLUSIONS: This first pneumococcal conjugate vaccine study in Asia confirms that the 11-valent diphtheria- and tetanus-conjugated pneumococcal vaccine is highly immunogenic in Filipino infants. The GMCs against most pneumococcal serotypes were substantially higher than described with the same or other pneumococcal conjugate vaccines in other populations. PMID- 12075763 TI - Reduced effectiveness of Haemophilus influenzae type b conjugate vaccine in children with a high prevalence of human immunodeficiency virus type 1 infection. AB - BACKGROUND: Haemophilus influenzae type b (Hib) conjugate vaccines have successfully reduced the burden of invasive Hib disease in developed countries; however, their effectiveness in countries with a high incidence of pediatric HIV 1 is unknown. METHODS: The effectiveness of Hib conjugate vaccine was prospectively evaluated in South African children. The burden of invasive Hib disease in children < 1 year old was compared in 2 cohorts. The first cohort included 22,000 African children born in 1997 [969 (4.45%) of whom were estimated to be HIV-1-infected] who were not vaccinated with Hib conjugate vaccine. This group was compared with 19,267 children [1162 (6.03%) of whom were estimated to be HIV-1 infected] vaccinated at 6, 10 and 14 weeks of age with an Hib conjugate vaccine [TETRAMUNE (polyribosylribitol phosphate-CRM(197)-diphtheria-tetanus toxoids-whole cell pertussis)] between March, 1998, and June, 1999. RESULTS: The estimated burden of invasive Hib disease in nonimmunized HIV-1-infected children < 1 year of age was 5.9-fold [95% confidence interval (95% CI), 2.7 to 12.6] higher than in HIV-1-uninfected children. The overall estimated effectiveness of Hib conjugate vaccine in fully vaccinated children <1 year of age was 83.2% (95% CI 60.3 to 92.9). Vaccine effectiveness was significantly reduced in HIV-1 infected [43.9% (95% CI -76.1 to 82.1)] compared with uninfected children [96.5% (95% CI 74.4 to 99.5); P < 10(-5)]. Among three of the fully vaccinated HIV-1 infected children who developed invasive Hib disease, the anti-Hib polyribosylribitol phosphate serum antibody concentrations were 0.23, 0.25 and 0.68 microg/ml. CONCLUSION: Although the Hib conjugate vaccine was less effective among HIV-1-infected than among uninfected children, it was 83% effective in preventing overall invasive Hib disease and therefore should be considered for inclusion in the routine vaccination schedule by other African countries. PMID- 12075764 TI - Introductory evaluation of an oral, killed whole cell enterotoxigenic Escherichia coli plus cholera toxin B subunit vaccine in Egyptian infants. AB - BACKGROUND: We conducted the first trial to assess the safety and immunogenicity of an oral, killed enterotoxigenic Escherichia coli plus cholera toxin B-subunit vaccine in children <2 years old. METHODS: Three doses of vaccine or killed E. coli K-12 control were given at 2-week intervals to 64 Egyptian infants, 6 to 18 months old, in a randomized, double blind manner. Adverse events were monitored for 3 days after each dose. Blood was collected before immunization and 7 to 10 days after each dose to assess vaccine-specific serologic responses. RESULTS: There was no statistically significant intergroup difference in the percentage of subjects reporting the primary safety endpoint (diarrhea or vomiting) after the first (31%, vaccine; 30%, control) or third (14%, vaccine; 18%, control) dose, whereas there was a trend toward greater reporting in the vaccine group after Dose 2 (36%, vaccine; 18%, control; P = 0.052). The percentage of children showing IgA seroconversion after any dose was higher in the vaccine than the control group for recombinant cholera toxin B-subunit (97% vs. 46%), colonization factor antigen I (61% vs. 18%) and coli surface antigen 4 (39% vs. 4%) (P < 0.001 for each comparison). IgG seroconversion rates in the vaccine and control groups were 97 and 21% to recombinant cholera toxin B-subunit (P < 0.001), 64 and 29% for colonization factor antigen I (P < 0.01), 53 and 21% for coli surface antigen 2 (P < 0.05) and 58 and 4% for coli surface antigen 4 (P < 0.001), respectively. The third vaccine dose was followed by augmented IgG antitoxin titers. CONCLUSION: The oral enterotoxigenic E. coli vaccine was safe and immunogenic in this setting in Egyptian infants. PMID- 12075765 TI - Low serum cortisol in combination with high adrenocorticotrophic hormone concentrations are associated with poor outcome in children with severe meningococcal disease. AB - OBJECTIVES: To study the correlation between serum concentrations of adrenocorticotrophic hormone (ACTH) and cortisol in relation to severity of disease in children with meningococcal sepsis. METHODS: Subjects were children with meningococcal sepsis, admitted to the pediatric intensive care unit. Clinical data, laboratory values and blood samples were selected. Arterial cortisol, ACTH, interleukin 6 and tumor necrosis factor alpha concentrations were measured on admission and studied for their relation to severity of disease (sepsis, septic shock/survivors, septic shock/nonsurvivors). RESULTS: Seventy-two patients fulfilled the criteria for meningococcal sepsis. Sixty-two of these children with positive blood cultures of Neisseria meningitidis, who were not treated with corticosteroids before admission, were included. Fifty of the 62 patients had septic shock. Twelve of those children (24%) died. The median age of the subjects was 2.6 years (range, 0.3 to 16.1 years). Cortisol values were significantly lower in non-survivors (median, 654 nmol/l) than in survivors (median, 2184 nmol/l) (P < 0.01). ACTH values were significantly higher in children who died (median, 1271 ng/l) than in survivors (85 ng/l) (P < 0.01). The median cortisol:ACTH ratio decreased significantly depending on the disease severity categories. CONCLUSIONS: Low serum cortisol concentrations in combination with high ACTH concentrations are associated with poor outcome in children with severe meningococcal disease. PMID- 12075766 TI - Inverse relationship between six week postvaccination varicella antibody response to vaccine and likelihood of long term breakthrough infection. AB - BACKGROUND: We used the large clinical database that supported the development of Oka/ Merck varicella vaccine to study the relationship between the primary varicella antibody response, as determined by gpELISA, an enzyme-linked immunosorbent assay that detects antibodies to varicella-zoster virus (VZV) glycoprotein, and the subsequent risk of postvaccination breakthrough varicella. METHODS: We vaccinated 1,164 healthy children with a single dose of varicella vaccine containing 2900 to 9000 plaque-forming units/dose. The primary immune response to vaccination was determined by gpELISA 6 weeks after vaccination. Subjects were followed annually for 7 years to ascertain cases of breakthrough varicella. RESULTS: The estimated vaccine efficacy among children with a 6-week postvaccination antibody titer of > or = 5 gpELISA units was 95.5% (95% confidence interval, 94.2%, 96.8%) compared with 83.5% (95% confidence interval, 76.9%, 89.5%) for subjects with a titer of <5 gpELISA units. Children with a 6 week postvaccination antibody titer of <5 gpELISA units were 3.5 times more likely than those with a titer of > or = 5 gpELISA units to develop breakthrough varicella. CONCLUSIONS: We identified a 6-week postvaccination antibody titer of > or = 5 gpELISA units as an approximate correlate of protection. In addition we established an accelerated failure time model based on log normal hazard that predicted varicella breakthrough rates based on the distribution of 6-week postvaccination varicella antibody titers. PMID- 12075767 TI - Alpha-hemolytic streptococcal infections among immunocompromised hosts: increasing incidence, severity and antibiotic resistance. PMID- 12075768 TI - Fluoroquinolones: considerations for future use. PMID- 12075769 TI - Kinetics of maternal hepatitis a antibody decay in infants: implications for vaccine use. AB - We conducted a seroepidemiologic study to evaluate the kinetics of maternal hepatitis A antibody decay in infants. Serum samples obtained from 200 infants at 2 and 4 months of age were tested for hepatitis A antibody. Seventy-six infants (38%) were hepatitis A antibody-positive with a geometric mean antibody titer of 2634 mIU/ml. Samples collected at 4, 6 and/or 12 months of age showed seropositivity rates of 100, 95 and 39%, respectively. These data indicate that maternal antibody levels remained high through the first 6 months of life but decayed significantly by 12 months of age. PMID- 12075770 TI - Population-based, age-specific myringotomy with tympanostomy tube insertion rates in Calgary, Canada. AB - Population-based rates of myringotomy with tympanostomy tube insertions in Calgary from 1997 to 2000 were determined with the use of administrative databases. Age-specific rates (procedures per 1,000 people per year) were 12.8, 54.2 and 11.1 for ages 0 to 11 months, 12 to 23 months and 0 to 15 years, respectively. Repeated procedures were performed on 7.1% of children <5 years of age. PMID- 12075771 TI - Safety, tolerability and immunogenicity of low dose Haemophilus influenzae type b conjugated to the outer membrane protein complex of Neisseria meningitidis group B. AB - Infants vaccinated with three doses followed by a booster of either 1.0 microg of Haemophilus influenzae type b conjugated to the outer membrane protein complex of Neisseria meningitidis group B or H. influenzae 10-microg oligosaccharide vaccine conjugated to 25 microg mutant toxin (CRM197) isolated from Corynebacterium diphtheriae had protective antipolyribosylribitol phosphate concentrations. PMID- 12075772 TI - Polyarthritis and humeral epiphysial separation in an infant with acute disseminated histoplasmosis. AB - The case of an immunocompetent infant with disseminated histoplasmosis is described. The case is unusual in its clinical presentation in that it is dominated, apart from respiratory infection, by the presence of polyarthritis and complicated by epiphysial separation of both humeri. There was only little involvement of reticuloendothelial tissues. Treatment consisted of surgical correction and itraconazole. PMID- 12075773 TI - Cerebellar ataxia as the presenting manifestation of Lyme disease. AB - A 7-year-old boy from suburban Baltimore who presented with cerebellar ataxia and headaches was found by magnetic resonance imaging to have multiple cerebellar enhancing lesions. He had no history of tick exposure. He was initially treated with steroids for presumptive postinfectious encephalitis. Lyme disease was diagnosed 10 weeks later after arthritis developed. Testing of the cerebrospinal fluid obtained at the time cerebellar ataxia was diagnosed revealed intrathecal antibody production to Borrelia burgdorferi. Treatment with intravenous antibiotics led to rapid resolution of persistent cerebellar findings. PMID- 12075774 TI - Peripheral thromboembolism associated with Malassezia furfur sepsis. AB - Malassezia furfur fungemia can cause sepsis in low birth weight neonates receiving parenteral lipids through central intravenous catheters. Its presentation has varied from nonspecific signs and symptoms to pulmonary vasculitis and endocarditis. We report the case of a premature infant who developed peripheral thromboembolic phenomena without evidence of endocarditis associated with M. furfur fungemia, an association not previously described. PMID- 12075775 TI - Chronic maternal hepatitis B infection and premature rupture of membranes. PMID- 12075776 TI - Erythema migrans associated with intravenous cephalosporin therapy. PMID- 12075777 TI - Dose of Calmette-Guerin bacillus vaccination in infants. PMID- 12075778 TI - Speciation of gas-phase and fine particle emissions from burning of foliar fuels. AB - Fine particle matter with aerodynamic diameter <2.5 microm (PM2.5) and gas-phase emissions from open burning of six fine (foliar) fuels common to fire-prone U.S. ecosystems are investigated. PM2.5 distribution is unimodal within the 10-450 nm range, indicative of an accumulation mode. Smoldering relative to flaming combustion shows smaller particle number density per unit time and median size. Over 100 individual organic compounds in the primarily carbonaceous (>70% by mass) PM2.5 are chemically speciated by gas chromatography/mass spectrometry. Expressed as a percent of PM2.5 mass, emission ranges by organic compound class are as follows: n-alkane (0.1-2%), polycyclic aromatic hydrocarbon (PAH) (0.02 0.2%), n-alkanoic acid (1-3%), n-alkanedioic acid (0.06-0.3%), n-alkenoic acid (0.3-3%), resin acid (0.5-6%), triterpenoid (0.2-0.5%), methoxyphenol (0.5-3%), and phytosterol (0.2-0.6%). A molecular tracer of biomass combustion, the sugar levoglucosan is abundant and constitutes a remarkably narrow PM2.5 mass range (2.8-3.6%). Organic chemical signatures in PM2.5 from open combustion of fine fuels differ with those of residential wood combustion and other related sources, making them functional for source-receptor modeling of PM. Inorganic matter [PM2.5 - (organic compounds + elemental carbon)] on average is estimated to make up 8% of the PM2.5. Wavelength dispersive X-ray fluorescence spectroscopy and ion chromatography identify 3% of PM2.5 as elements and water-soluble ions, respectively. Compared with residential wood burning, the PM2.5 of fine fuel combustion is nitrate enriched but shows lower potassium levels. Gas-phase C2-C13 hydrocarbon and C2-C9 carbonyl emissions are speciated by respective EPA Methods T0-15 and T0-11A. They comprise mainly low molecular weight C2-C3 compounds and hazardous air pollutants (48 wt % of total quantified volatile organic carbon). PMID- 12075779 TI - Methyl triclosan found in Swiss lakes. PMID- 12075780 TI - Black carbon from the Mississippi River: quantities, sources, and potential implications for the global carbon cycle. AB - Black carbon (BC) may be a major component of riverine carbon exported to the ocean, but its flux from large rivers is unknown. Furthermore, the global distribution of BC between natural and anthropogenic sources remains uncertain. We have determined BC concentrations in suspended sediments of the Mississippi River, the 7th largest river in the world in terms of sediment and water discharge, during high flow and low flow in 1999. The 1999 annual flux of BC from the Mississippi River was 5 x 10(-4) petagrams (1 Pg = 10(15) g = 1 gigaton). We also applied a principal components analysis to particulate-phase high molecular weight polycyclic aromatic hydrocarbon isomer ratios in Mississippi River suspended sediments. In doing so, we determined that approximately 27% of the BC discharged from the Mississippi River in 1999 originated from fossil fuel combustion (coal and smelter-derived combustion), implicating fluvial BC as an important source of anthropogenic BC contamination into the ocean. Using our value for BC flux and the annual estimate for BC burial in ocean sediments, we calculate that, in 1999, the Mississippi River discharged approximately 5% of the BC buried annually in the ocean. These results have important implications, not only for the global carbon cycle but also for the fluvial discharge of particulate organic contaminants into the world's oceans. PMID- 12075782 TI - Putting regulations under a microscope. PMID- 12075781 TI - Atmospherc mercury deposition during the last 270 years: a glacial ice core record of natural and anthropogenic sources. AB - Mercury (Hg) contamination of aquatic ecosystems and subsequent methylmercury bioaccumulation are significant environmental problems of global extent. At regional to global scales, the primary mechanism of Hg contamination is atmospheric Hg transport. Thus, a better understanding of the long-term history of atmospheric Hg cycling and quantification of the sources is critical for assessing the regional and global impact of anthropogenic Hg emissions. Ice cores collected from the Upper Fremont Glacier (UFG), Wyoming, contain a high resolution record of total atmospheric Hg deposition (ca. 1720-1993). Total Hg in 97 ice-core samples was determined with trace-metal clean handling methods and low-level analytical procedures to reconstruct the first and most comprehensive atmospheric Hg deposition record of its kind yet available from North America. The record indicates major atmospheric releases of both natural and anthropogenic Hg from regional and global sources. Integrated over the past 270-year ice-core history, anthropogenic inputs contributed 52%, volcanic events 6%, and background sources 42%. More significantly, during the last 100 years, anthropogenic sources contributed 70% of the total Hg input. Unlike the 2-7-fold increase observed from preindustrial times (before 1840) to the mid-1980s in sediment-core records, the UFG record indicates a 20-fold increase for the same period. The sediment-core records, however, are in agreement with the last 10 years of this ice-core record, indicating declines in atmospheric Hg deposition. PMID- 12075783 TI - Plasma biomarkers in fish provide evidence for endocrine modulation in the Elbe River, Germany. AB - Blood plasma samples were collected from wild bream (Abramis brama L.) in the Elbe River, Germany, and analyzed for the yolk protein precursor vitellogenin (VTG), a biomarker for estrogen exposure, and the sex steroids 11 ketotestosterone (11KT), testosterone (T), and 17beta-estradiol (E2) to investigate for evidence of endocrine modulation. In addition, the gonadal status and the prominence of spawning tubercles were investigated. Nine riverine sites were investigated on the Elbe that were influenced by different sources of endocrine-active substances. Bream were collected from a lake that received no domestic or industrial discharges as a control. Plasma VTG concentrations were significantly higher in male bream from the Czech border to the middle Elbe, with the highest concentrations in fish sampled at the locations near Magdeburg and downstream of Dresden (between 20 and 100 times higher than in the controls), regions that are characterized by high levels of effluent discharges into the river. Following the Elbe from this site to the sea, the concentrations of plasma VTG in males were lower than at Meissen but were still elevated above the controls. 11KT and E2 titers showed suppressions in their normal concentrations at some locations (those receiving the greatest industrial discharges). There were reciprocal relationships between inhibitory effects on gonadal growth, maturation, and plasma sex steroids and exposure to pollutants, such as organotins, pesticides, or metals. However, there was no single chemical that alone could explain the observed inhibitory effects on sexual development. The results indicate that the endocrine system in wild bream is disrupted in stretches of the Elbe River. PMID- 12075784 TI - Ice cores open new window on historical mercury deposition. PMID- 12075786 TI - Potential remobilization of 137Cs, 60Co, 90Tc, and 90Sr from contaminated Mayak sediments in river and estuary environments. AB - Following 50 years of nuclear production at Mayak PA, sediments in storage reservoirs are significantly contaminated. Dam failure or flooding could potentially transport large amounts of sediments, via rivers, to the Ob estuary and Kara Sea. The objectives of this work were to investigate fresh and seawater remobilization of 137Cs, 50Co, 99Tc, and 90Sr from contaminated Reservoir 10 sediments. Sediments were extracted sequentially using synthetic Techa freshwater, seawater, and chemical reagents with increasing dissolution powers. 137Cs and 90Sr freshwater distribution coefficients (apparent Kd) agreed quite well with published values; values for 99Tc were higher and values for 60Co were lower than expected. In seawater, mean apparent Kd values decreased by 94, 77, 48, and 73% (137Cs, 60Co, 99Tc, and 90Sr, respectively), indicating increased radionuclide mobility. Remobilization in seawater was 5, 15, 1, and 23% of total activities (i.e., releases of 165, 11, 0.3, and 170 kBq kg(-1) d.w.) for 137Cs, 60Co, 99Tc, and 90Sr, respectively. 137Cs and 99Tc were strongly bound to sediments (60% and 80%, respectively). 60Co and 90Sr were more mobile (70% reversibly bound). In conclusion, Mayak Reservoir sediments could potentially contaminate the Ob estuary due to remobilization of sediment-held radionuclides upon contact with seawater. PMID- 12075785 TI - Occurrence and environmental behavior of the bactericide triclosan and its methyl derivative in surface waters and in wastewater. AB - The bactericide triclosan and methyl triclosan, an environmental transformation product thereof, were detected in lakes and in a river in Switzerland at concentrations of up to 74 and 2 ng L(-1), respectively. Both compounds were emitted via wastewater treatment plants (WWTPs), with methyl triclosan probably being formed by biological methylation. A regional mass balance for a lake (Greifensee) indicated significant removal of triclosan by processes other than flushing. Laboratory experiments showed that triclosan in the dissociated form was rapidly decomposed in lake water when exposed to sunlight (half-life less than 1 h in August at 47 degrees latitude). Methyl triclosan and nondissociated triclosan, however, were relatively stable toward photodegradation. Modeling these experimental data for the situation of lake Greifensee indicated that photodegradation can account for the elimination of triclosan from the lake and suggested a seasonal dependence of the concentrations (lower in summer, higher in winter), consistent with observed concentrations. Although emissions of methyl triclosan from WWTPs were only approximately 2% relative to those of triclosan, its predicted concentration relative to triclosan in the epilimnion of the lake increases to 30% in summer. Passive sampling with semipermeable membrane devices (SPMDs) indicated the presence of methyl triclosan in lakes with inputs from anthropogenic sources but not in a remote mountain lake. Surprisingly, no parent triclosan was observed in the SPMDs from these lakes. Methyl triclosan appears to be preferentially accumulated in SPMDs under the conditions in these lakes, leading to concentrations comparable to those of persistent chlorinated organic pollutants. PMID- 12075787 TI - Distribution and reactivity of O2-reducing components in sediments from a layered aquifer. AB - The redox status of subsurface aqueous systems is controlled by the reactivity of solid redox-sensitive species and by the inflow of such species dissolved in groundwater. The reactivity toward molecular oxygen (O2) of solid reductants present in three particle size fractions of sediments from a pristine aquifer was characterized during 54 days. The stoichiometric relationships between carbon dioxide (CO2) production and O2 consumption was used in combination with sulfate production to discriminate between the contributions of sedimentary organic matter (0-87%), pyrite (6-100%), and siderite (0-43%) as the dominant reductants. The observed simultaneous oxidation of these reductants indicates that they are reactive on the same time scales. The measured reduction capacity 18-84 micromol O2/g) ranged from 8 to 42% of the total reduction capacity present as pyrite and organic carbon in the total sediment fraction (<2 mm). Fine fractions (<63 microm) were 10-250 times more reactive than their corresponding total fractions. Oxygen consumption rates decreased continuously during carbonate buffered conditions, due to a decreasing reactivity of reductants. Acidification accelerated pyrite oxidation but impeded SOM respiration. Our findings indicate that the geological history of aquifer sediments affects the amounts of organic matter, pyrite and siderite present, while environmental conditions, such as pH and microbial activity, are important in controlling the reactivity of these reductants. These controls should be considered when assessing the natural reduction activity of aquifer sediments in either natural or polluted systems. PMID- 12075788 TI - Uptake of dioxins by pastures more rapid than previously thought. PMID- 12075789 TI - Effects of meteorological conditions on aerosol composition and mixing state in Bakersfield, CA. AB - Particle and meteorological instrumentation were used to characterize ambient atmospheric conditions, aerosol size distributions, aerosol mass concentrations, and single particle size and chemical composition in Bakersfield, CA for the period January 9, 1999 through January 28, 1999. The sampling period included four distinct meteorological periods of stagnation, clearing, haze, and rain. Particle number and mass concentrations were the highest during the stagnation episode when a heavy and extensive fog developed. Mass and number concentrations also approached these high levels during the haze period. Single particle size and composition data from an aerosol time-of-flight mass spectrometer (ATOFMS) are used to provide unique continuous information on the diversity in types of particles present, the effects of meteorology on particle size and composition, and the distribution of important chemical species within individual particles. Aerosol composition and mixing state are found to vary with meteorological conditions. Single particle data show that carbonaceous aerosol with secondary ammonium, nitrate, and sulfate dominate the aerosol concentration during a stagnation period with a dramatic composition shift occurring to sodium type particles during the haze period. The aerosol is internally mixed with respect to carbon, nitrate, sulfate, and ammonium during the stagnation period. The mixing state changes significantly over the haze period when much greater diversity in the associations of chemical species within individual particles occurs. PMID- 12075790 TI - Reassessment of the hydrocarbons in Prince William Sound and the Gulf of Alaska: identifying the source using partial least-squares. AB - There has been considerable exchange in the literature regarding the source of the background hydrocarbons in Prince William Sound and the Gulf of Alaska. Page and coworkers suggest oil-based sources while Short and coworkers suggest coal. The multivariate statistical methodology of partial least-squares (PLS) has been used to reassess the percentage contribution of coal, seep oil, shales, and rivers to the hydrocarbon loading in the Gulf of Alaska. Data have kindly been provided by Short (NOAA) and Page (Bowdoin College, for Exxon), and these have been analyzed using selected sites as sources in order to develop signatures. These signatures are based on 40 and 136 compounds, respectively, and include the PAH and terpane/sterane biomarkers in the case of the Exxon data. The principal components describing these sources are then fitted to the data for other sites around the Prince William Sound (PWS) and Gulf of Alaska (GoA) to determine the proportion of the variability described by each source. Using the Exxon data, a mixed source of coal, seep oil, eroding shales, and rivers (1 and 2) sources described approximately 13%, 18%, 24%, 26%, and 20%, respectively, of the variance in PWS and GoA data. The rivers 1 signature was very similar to that of coal, while rivers 2 was more similar to the eroding shales. New coal data (Short, unpublished work) also indicated considerable overlap with the Exxon seep oil. With the NOAA data, spatial plots of the explained variance indicate that the prespill background has a wide range of explained fits. There is considerable overlap in the signatures developed from the data, and Coomans' Plots identify those compounds which are the most diagnostic: for the Exxon seep oil signature, naphthalene and methyl- and dimethylnaphthalene are the best markers, whereas for the NOAA prespill background, Exxon coals and shales are best defined by the larger PAHs such as benzo[ghi]perylene. The evidence suggests mixed sources whose contributions vary significantly across the sampling area. PMID- 12075791 TI - Source apportionment of PM2.5 in the Southeastern United States using solvent extractable organic compounds as tracers. AB - A chemical mass balance (CMB) receptor model using particle-phase organic compounds as tracers is applied to apportion the primary source contributions to fine particulate matter and fine particulate organic carbon concentrations in the southeastern United States to determine the seasonal variability of these concentrations. Source contributions to particles with aerodynamic diameter < or =2.5 microm (PM2.5) collected from four urban and four rural/suburban sites in AL, FL, GA, and MS during April, July, and October 1999 and January 2000 are calculated and presented. Organic compounds in monthly composite samples at each site are identified and quantified by gas chromatography/mass spectrometry and are used as molecular markers in the CMB model. The major contributors to identified PM2.5 organic carbon concentrations at these sites in the southeastern United States include wood combustion (25-66%), diesel exhaust (14-30%), meat cooking (5-12%), and gasoline-powered motor vehicle exhaust (0-10%), as well as smaller but statistically significant contributions from natural gas combustion, paved road dust, and vegetative detritus. The primary sources determined in the present study when added to secondary aerosol formation account for on average 89% of PM2.5 mass concentrations, with the major contributors to PM2.5 mass as secondary sulfate (30+/-6%), wood combustion (15+/-12%), diesel exhaust (16+/ 7%), secondary ammonium (8+/-2%), secondary nitrate (4+/-3%), meat cooking (3+/ 2%), gasoline-powered motor vehicle exhaust (2+/-2%), and road dust (2+/-2%). Distinct seasonality is observed in source contributions, including higher contributions from wood combustion during the colder months of October and January. In addition, higher percentages of unexplained fine organic carbon concentrations are observed in July, which are likely due to an increase in secondary organic aerosol formation during the summer season. PMID- 12075792 TI - Polychlorinated dibenzo-p-dioxin and furan (PCDD/F) uptake by pasture. AB - Uptake of airborne PCDD/Fs by a native pasture sward was studied. The concentrations of the less chlorinated PCDD/Fs (up to and including the Cl5DD/Fs) in pasture harvested on the same day after 2, 6, and 12 weeks exposure were the same, implying that air-pasture steady-state was reached within 2 weeks of exposure. The implications of these observations for the relative importance of input (atmospheric deposition, soil resuspension) and loss (photolysis, degradation, volatilization, cuticular shedding, and growth dilution) processes are discussed and inferences made about the controlling factors. The concentrations of the more chlorinated PCDD/Fs were more variable. We infer that they were influenced by wash-off of particle- and surface-bound chemical. Up to 4% of the Cl2-4DD/F and 4-13% of the Cl5-8DD/F loading on the pasture was estimated to have been supplied by adhering soil particles, with the remainder supplied by atmospheric deposition. Between 0.04 and 0.66 kg of each homologue group (excluding Cl2DFs) is estimated to be transferred annually from the atmosphere to pasture in the U.K. PMID- 12075793 TI - Decadal trends of silver and lead contamination in San Francisco Bay surface waters. AB - Over the past decade, San Francisco Bay surface waters have remained enriched with dissolved (<0.45 microm) silver and lead concentrations (decadal means of 5.7 ng kg(-1) Ag[filtered] and 31 ng kg(-1) Pb[filtered]) compared with those (0.26 ng kg(-1) Ag[filtered] and 2.7 ng kg(-1) Pb[filtered]) of adjacent oceanic surface waters of the northeast Pacific, despite efforts to reduce pollutant loadings to the Bay during that period. While time series models show that there has been a 40% decline in total lead concentrations in the southern reach of the estuarine system between 1989 and 1999, the filtered lead fraction has not changed significantly during that time. That persistence is attributed to (i) the ongoing input from previous atmospheric depositions and industrial lead to its drainage basin, which are slowly being advected into the estuary and (ii) the internal recycling of lead between the surface sediments and the water column within the Bay. In contrast, both filtered and total silver concentrations in the southern reach have declined by 70% and 40%, respectively, within the past decade. These temporal declines are attributed to a 2-fold decrease in silver loadings from publicly owned treatment works and a comparable decline in the silver concentration of surficial sediments within that region during the past decade. In the northern reach, silver and lead concentrations have remained essentially constant between 1989 and 1999, reflecting invariable anthropogenic input of these elements to this embayment over that decade. PMID- 12075794 TI - Anthropogenic Gd in surface water, drainage system, and the water supply of the city of Prague, Czech Republic. AB - The anthropogenic Gd (Gdant) loads and fluxes of surface- and groundwaters, and of effluents from sewage treatment plants of Prague and its surroundings, have been determined. While passing Prague the Gdant load of the Vltava river increases from 4.5 to 19.2 nmol/m3, which is caused by the effluents of the central sewage treatment plant with a 6 day average of 250 nmol/m3. The Berounka river with its enhanced content doubles the Gdant flux of the Vltava river before entering Prague. All minor creeks draining Prague add less than 3% of the Gdant load in the Vltava river downstream of Prague. The 11-days average of the amount of the administered Gd complexes of the gadopentetic acid (Gd-DTPA), gadobenic acid, and other compounds in Prague's hospitals (22 g-Gdant/d) exceeds the 6 days average of 15 g-Gdant/d in the effluent of the central sewage treatment plant. This strongly indicates uncontrolled losses of sewage from sewers to the groundwater. PMID- 12075795 TI - Atmospheric chemistry of HFE-7500 [n-C3F7CF(OC2H5)CF(CF3)2]: reaction with OH radicals and Cl atoms and atmospheric fate of n-C3F7CF(OCHO*)CF(CF3)2 and n C3F7CF(OCH2CH2O*)CF(CF3)2 radicals. AB - Relative rate techniques were used to measure k(OH + HFE-7500) = (2.6+/-0.6) x 10(-14), k(Cl + HFE-7500) = (2.3+/-0.7) x 10(-12), k[Cl + n C3F7CF(OC(O)H)CF(CF3)2] = (9.7+/-1.4) x 10(-15), and k[Cl + n C3F7CF(OC(O)CH3)CF(CF3)2] < 6 x 10(-17) cm3 molecule(-1) s(-1) at 295 K [HFE-7500 = n-C3F7-CF(OC2H5)CF(CF3)2]. From the value of k(OH + HFE-7500) an estimate of 2.2 years for the atmospheric lifetime of HFE-7500 is obtained. Two competing loss mechanisms for n-C3F7-CF(OCHO.CH3)CF(CF3)2 radicals were identified in 700 Torr of N2/O2 diluent at 295 K; reaction with O2 and decomposition via C-C bond scission with kO2/k(decomp) = 0.013+/-0.006 Torr(-1). The Cl atom initiated oxidation of HFE-7500 in N2/O2 diluent gives n-C3F7CF(OC(O)CH3)CF(CF3)2 as the major product and n-C3F7CF(OC(O)H)CF(CF3)2 as a minor product. The atmospheric oxidation of HFE-7500 gives n-C3F7-CF(OC(O)CH3)CF(CF3)2 and n C3F7CF(OC(O)H)CF(CF3)2 as oxidation products. The results are discussed with respect to the atmospheric chemistry and environmental impact of HFE-7500. PMID- 12075796 TI - Field and laboratory investigations of inactivation of viruses (PRD1 and MS2) attached to iron oxide-coated quartz sand. AB - Field and laboratory experiments were conducted to investigate inactivation of viruses attached to mineral surfaces. In a natural gradient transport field experiment, bacteriophage PRD1, radiolabeled with 32P, was injected into a ferric oxyhydroxide-coated sand aquifer with bromide and linear alkylbenzene sulfonates. In a zone of the aquifer contaminated by secondary sewage infiltration, small fractions of infective and 32P-labeled PRD1 broke through with the bromide tracer,followed bythe slow release of 84% of the 32P activity and only 0.011% of the infective PRD1. In the laboratory experiments, the inactivation of PRD1, labeled with 35S (protein capsid), and MS2, dual radiolabeled with 35S (protein capsid) and 32P (nucleic acid), was monitored in the presence of groundwater and sediment from the contaminated zone of the field site. Release of infective viruses decreased at a much faster rate than release of the radiolabels, indicating that attached viruses were undergoing surface inactivation. Disparities between 32P and 35S release suggest that the inactivated viruses were released in a disintegrated state. Comparison of estimated solution and surface inactivation rates indicates solution inactivation is approximately 3 times as fast as surface inactivation. The actual rate of surface inactivation may be substantially underestimated owing to slow release of inactivated viruses. PMID- 12075797 TI - Safe storage of CO2 in deep saline aquifers. PMID- 12075798 TI - Emission of alcohols and carbonyl compounds from a spark ignition engine. Influence of fuel and air/fuel equivalence ratio. AB - A spark ignition engine was used to study the impact of fuel composition and of the air/fuel equivalence (lambda) ratio on exhaust emissions of alcohols and aldehydes/ketones. Fuel blends contained eight hydrocarbons (n-hexane, 1-hexene, cyclohexane, n-octane, isooctane, toluene, o-xylene, and ethylbenzene (ETB)) and four oxygenated compounds (methanol, ethanol, 2-propanol, and methyl tert butyl ether (MTBE)). Exhaust methanol is principally produced from fuel methanol and MTBE but also from ethanol, 2-propanol, isooctane, and hexane. Exhaust ethanol and 2-propanol are produced only from the respective fuel compounds. Exhaust formaldehyde is mainly produced from fuel methanol, acetaldehyde from fuel ethanol, and propionaldehyde from straight-chain hydrocarbons. Exhaust acroleine comes from fuel 1-hexene, acetone from 2-propanol, n-hexane, n-octane, isooctane, and MTBE. Exhaust crotonaldehyde comes from fuel 1-hexene, cyclohexane, n-hexane, and n-octane, methacroleine from fuel isooctane, and benzaldehyde from fuel aromatics. Light pollutants (C1-C2) are most likely formed from intermediate species which are quite independent of the fuel composition. An increase in A increases the exhaust concentration of acroleine, crotonaldehyde, methacroleine, and decreases these of the three alcohols for the alcohol-blended fuels. The concentration of methanol, formaldehyde, propionaldehyde, and benzaldehyde is a maximum atstoichiometry. The exhaust concentration of acetaldehyde and acetone presents a complex behavior: it increases in some cases, decreases in others, or presents a maximum at stoichiometry. The concentration of four aldehydes (formaldehyde, acetaldehyde, propionaldehyde, and benzaldehyde) is also linked with the exhaust temperature and fuel H/C ratio. PMID- 12075799 TI - The effects of organic species on the hygroscopic behaviors of inorganic aerosols. AB - Water-soluble organic compounds have recently received much attention because of their ability to absorb water and alter the hygroscopic properties of inorganic aerosols. The effects of glycerol, succinic acid, malonic acid, citric acid, and glutaric acid on the water cycles (water activities during particle evaporation and growth), crystallization relative humidities (CRH), and deliquescence relative humidities (DRH) of sodium chloride (NaCl) and ammonium sulfate (AS) were studied using an electrodynamic balance (EDB). The growth factors of these inorganic and organic mixtures were lower than those of the pure inorganic species. The presence of all these organics in the mixed particle reduce the water absorption of NaCl but enhance that of AS relative to that of the pure inorganic salts. Glycerol and succinic acid did not affect the deliquescence properties of NaCl and AS, although succinic acid increased the CRH of NaCl and AS. Malonic acid and citric acid, behaving as nondeliquescent species in single particle studies, caused NaCl and AS particles to absorb a significant amount of water before deliquescence. Glutaric acid caused NaCl and AS to deliquesce gradually, spanning a wide range of relative humidity. The ZSR model was found to be useful in predicting the water activity of the mixtures and the growth ratios. However, the detailed crystallization and deliquescence behaviors of the organic/inorganic mixtures cannot be easily predicted from the hygroscopic properties of the individual components. PMID- 12075800 TI - Soil ecotoxicity of polycyclic aromatic hydrocarbons in relation to soil sorption, lipophilicity, and water solubility. AB - A data set was generated aiming to predict the toxicity of PAHs to soil organisms. Toxicity data include the effects of 16 PAHs on the survival and reproduction of the soil-dwelling springtail Folsomia fimetaria. The results show that only PAHs with reported log Kow values < or = 5.2 (i.e., naphthalene, acenaphthene, acenaphthylene, anthracene, phenanthrene, fluorene, pyrene, and fluoranthene) significantly affected the survival or reproduction of the test organisms. Threshold values for the toxicity of the individual PAHs could be expressed as pore-water concentrations by the use of reported organic carbon normalized soil-pore-water partitioning coefficients (Koc values). For the PAHs with a log Kow < or = 5.2, toxicity significantly increased with increasing lipophilicity of the substances (r2 = 0.67; p = 0.012; n = 8), suggesting a narcotic mode of toxic action for most substances. However, the position of anthracene in the regression plot indicated a more specific mode of toxic action than narcosis, and removing this data point yielded the following regression equation: log EC10 (micromol/L) = -0.97 log Kow + 4.0 (r2 = 0.80; p = 0.006; n = 7). Using this quantitative structure-activity relationship (QSAR) to calculate threshold values for the toxicity of the remaining nontoxic substances (benz[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, dibenz[a,h]anthracene, benzo[a]pyrene, perylene, and indeno[1,2,3-cd]pyrene), the absence of toxicity could, in most cases, be explained by a limited water solubility, indicating that these substances do act by narcosis as the mode of toxic action and that their toxicity is governed by concentrations in the pore water. The results provide important input to future model predictions of the ecological risk posed by PAH contaminated sites. PMID- 12075801 TI - Anaerobic methane oxidation in a landfill-leachate plume. AB - The alluvial aquifer adjacent to Norman Landfill, OK, provides an excellent natural laboratory for the study of anaerobic processes impacting landfill leachate contaminated aquifers. We collected groundwaters from a transect of seven multilevel wells ranging in depth from 1.3 to 11 m that were oriented parallel to the flow path. The center of the leachate plume was characterized by (1) high alkalinity and elevated concentrations of total dissolved organic carbon, reduced iron, and methane, and (2) negligible oxygen, nitrate, and sulfate concentrations. Methane concentrations and stable carbon isotope (delta13C) values suggest anaerobic methane oxidation was occurring within the plume and at its margins. Methane delta13C values increased from about -54 per thousand near the source to > -10 per thousand downgradient and at the plume margins. The isotopic fractionation associated with this methane oxidation was 13.6+/-1.0 per thousand. Methane 13C enrichment indicated that 80-90% of the original landfill methane was oxidized over the 210-m transect. First-order rate constants ranged from 0.06 to 0.23 per year, and oxidation rates ranged from 18 to 230 microM/y. Overall, hydrochemical data suggest that a sulfate reducer methanogen consortium may mediate this methane oxidation. These results demonstrate that natural attenuation through anaerobic methane oxidation can be an important sink for landfill methane in aquifer systems. PMID- 12075802 TI - Sediment-associated reactions of aromatic amines. 2. QSAR development. AB - The fate of aromatic amines in soils and sediments is dominated by irreversible binding through nucleophilic addition and oxidative radical coupling. Despite the common occurrence of the aromatic amine functional group in organic chemicals, the molecular properties useful for predicting reaction kinetics in natural systems have not been thoroughly investigated. Toward this goal, the sorption kinetics for a series of anilines with substituents in the ortho, meta, or para positions were measured in sediment slurries. The sorption kinetics of the substituted anilines were characterized by an initial, rapid sorption process followed by a much slower sorption process. The initial rates of sorption varied with the type and position of the substituent group. Rate constants for the initial sorption process were correlated with molecular descriptors, including dissociation constants (pKa's), Hammett sigma constants, polarographic half-wave potentials (E1/2), one-electron oxidation potentials (E1), highest occupied molecular orbital (HOMO) energies (E(HOMO)), and ionization energies (E(IE)). On the basis of the strength of linear correlations and the availability of data, dissociation constants and Hammett sigma constants appear to be the most useful molecular descriptors for predicting reaction rates of substituted anilines in the sediment slurries. The slow rates of sorption were much less sensitive to substituents effects than the rate constants for the faster sorption process, suggesting that the slower process was not controlled by the rate of electron transfer (i.e., nucleophilic addition or radical formation) but was limited by the availability of covalent binding sites. PMID- 12075803 TI - Raman study of aluminum speciation in simulated alkaline nuclear waste. AB - The chemistry of concentrated sodium aluminate solutions stored in many of the large, underground storage tanks containing high-level waste (HLW) at the Hanford and Savannah River Nuclear Reservations is an area of recent research interest. Not only is the presence of aluminate in solution important for continued safe storage of these wastes, the nature of both solid and solution aluminum oxyhydroxides is important for waste pretreatment. Moreover, for many tanks that have leaked high aluminum waste in the past, little is known about the speciation of Al in the soil. In this study, Raman spectroscopy has been used to investigate the speciation of the aqueous species in the Al2O3-Na2O-H2O system over a wide range of solution compositions and hydration. A ternary phase diagram has been used to correlate the observed changes in the spectra with the composition of the solution and with dimerization of aluminate that occurs at elevated aluminate concentrations (>1.5 M). Dimerization is evidenced by growth of new Al-O stretching bands at 535 and 695 cm(-1) at the expense of the aluminate monomer band at 620 cm(-1). The spectrum of water was strongly influenced by the high concentrations of Na+ and OH- (>17 M). Upon increasing the concentration of NaOH in solution, the delta-(H-O-H) bending band of water (v2 mode) increased in frequency to 1663 cm(-1), indicating that the water contained in the concentrated caustic solution was more strongly hydrogen bonded at the higher base content. In addition, the sharp, well-resolved band at 3610 cm(-1), assigned to the v(O-H) of free OH-, increased in intensity with increasing NaOH. Analysis of the v(O-H) bands in the 3800-2600 cm(-1) region supported the overall increase in hydrogen bonding as evidenced by the increase in relative intensity of a strongly hydrated water band at 3118 cm(-1). Taking into consideration the activity of water, the molar concentrations of the monomeric and dimeric aluminate species were estimated using the relative intensities of the Al-O stretching bands from the Raman spectra. A constant apparent log Kdimer value was obtained at aluminate concentrations >1.5 M with a value of 0.97+/-0.04 at approximately 25 degrees C. This study represents the first spectral-based estimation of a thermodynamic equilibrium constant for the Al2O3-Na2O-H2O system. PMID- 12075804 TI - Rates of hydrous ferric oxide crystallization and the influence on coprecipitated arsenate. AB - Arsenate coprecipitated with hydrous ferric oxide (HFO) was stabilized against dissolution during transformation of HFO to more crystalline iron (hydr)oxides. The rate of arsenate stabilization approximately coincided with the rate of HFO transformation at pH 6 and 40 degrees C. Comparison of extraction data and X-ray diffraction results confirmed that hematite and goethite were the primary transformation products. HFO transformation was significantly retarded at or above an arsenate solid loading of 29 455 mg As/kg HFO. However, HFO transformation proceeded at a significant rate for arsenate solid loadings of 4208 and 8416 mg As/kg HFO. At a solid loading of 8416 mg As/kg HFO, XRD results suggested arsenate primarily partitioned to hematite. Comparison of HFO transformation rates observed in this research to rates obtained from the literature at pH 6 and temperatures ranging from 24 to 70 degrees C suggests that arsenate stabilization could be realized in oxic environments with a significantfraction of iron (hydr)oxides. While this process has not been documented in natural systems, the predicted half-life for transformation of an arsenic-bearing HFO is approximately 300 days at 25 degrees C at solid loading < 8415 mg As/kg HFO. The projected time frame for arsenate stabilization indicates this process should be considered during development of conceptual and analytical models describing arsenic fate and transport in oxic systems containing reactive iron (hydr)oxides. The likelihood of this process would depend on the chemical dynamics of the soil or sediment system relative to iron (hydr)oxide precipitation-dissolution reactions and the potential retarding/competing influence of ions such as silicate and organic matter. PMID- 12075805 TI - Hydrogen isotopic enrichment: an indicator of biodegradation at a petroleum hydrocarbon contaminated field site. AB - Compound-specific carbon and hydrogen isotope analysis was used to investigate biodegradation of benzene and ethylbenzene in contaminated groundwater at Dow Benelux BV industrial site. delta13C values for dissolved benzene and ethylbenzene in downgradient samples were enriched by up to 2+/-0.5 per thousand, in 13C, compared to the delta13C value of the source area samples. delta2H values for dissolved benzene and ethylbenzene in downgradient samples exhibited larger isotopic enrichments of up to 27+/-5 per thousand for benzene and up to 50+/-5 per thousand for ethylbenzene relative to the source area. The observed carbon and hydrogen isotopic fractionation in downgradient samples provides evidence of biodegradation of both benzene and ethylbenzene within the study area at Dow Benelux BV. The estimated extents of biodegradation of benzene derived from carbon and hydrogen isotopic compositions for each sample are in agreement, supporting the conclusion that biodegradation is the primary control on the observed differences in carbon and hydrogen isotope values. Combined carbon and hydrogen isotope analyses provides the ability to compare biodegradation in the field based on two different parameters, and hence provides a stronger basis for assessment of biodegradation of petroleum hydrocarbon contaminants. PMID- 12075806 TI - Water quality security. PMID- 12075807 TI - Uptake of dissolved Ag, Cd, and Co by the clam, Macoma balthica: relative importance of overlying water, oxic pore water, and burrow water. AB - The facultative deposit-feeding clam Macoma balthica is used as a bioindicator organism for assessing coastal metal contamination. Previous work has evaluated the assimilation of metals from different possible food sources for this clam, but no studies have measured the uptake rates of metals from different dissolved sources. This study specifically compares three different dissolved sources: overlying water (SW), oxic pore water (OPW) from a depth of <1 cm (entrained during surface deposit feeding), and burrow water (BW) (a mixture of anoxic pore water and overlying water). Uptake rates of dissolved Ag, Cd, and Co in M. balthica were measured in short-term laboratory experiments using radiotracers. Clams were exposed to metals in water only for SW and surface OPW treatments. In the BW treatment, metal uptake was compared in clams placed in radiolabeled organic-poor or organic-rich sediment under conditions in which feeding was inhibited. Uptake rate constantsfrom SW for Ag, Cd, and Co were 0.35, 0.033, and 0.035 L g(-1) day(-1), respectively. Lower uptake of dissolved metals from OPW was noted but was only significant for Co. Metal uptake from BW and SW were also comparable; however, the trend showed lower Ag and higher Co uptake from BW. Metal distributions and concentrations in the two radiolabeled sediments were affected by active irrigation of SW into the burrows; dissolved metal concentrations in BW were approximately 30% lower than that in the bulk pore water concentrations. In the organic-rich sediment, Cd and Ag partitioned more in the dissolved phase (<0.2 microm) and Co more in the particulate phase as compared with the organic-poor sediment. A sensitivity analysis using measured rate constants for uptake and a range of metal concentrations from field studies suggested that, under most conditions, uptake of dissolved Ag is primarily from OPW, Co is mostly from BW, and Cd uptake varies depending on its concentration in each compartment. Little Co or Ag is likely to be taken up from SW, whereas 20 50% of Cd may be accumulated from this source. Thus, SW, OPW, and BW are all potential sources of metals for M. balthica, and the relative importance of these sources differs among metals and is dependent on the dissolved metal concentrations in each compartment. PMID- 12075808 TI - Natural formation of vinyl chloride in the terrestrial environment. AB - Vinyl chloride is a highly reactive and toxic substance which is widely used in industry. It is the parent compound of poly(vinyl chloride) (PVC), one of the most important industrial polymers. Until now, it was thought that vinyl chloride found in the environment is exclusively man-made or results from the degradation of other anthropogenic substances, such as trichloroethylene and tetrachloroethylene. Here, we demonstrate that vinyl chloride also has natural sources. Soil air and ambient air from a rural area in Northern Germany were investigated for volatile chlorinated halocarbons. The concentrations of vinyl chloride in the soil air were significantly enhanced as compared to ambient air, indicating a natural formation of this compound in the soil. A series of laboratory experiments using different soils and model compounds was conducted, which clearly proved that vinyl chloride could be produced during soil processes. We propose that this highly reactive compound can be formed during the oxidative degradation of organic matter in soil, for example, in a reaction between humic substances, chloride ions and an oxidant (ferric ions or hydroxyl radicals). The redox-sensitive aromatic compounds in soil such as catechols and o-quinones can be degraded to CO2, accompanied by the release of vinyl chloride and other volatile chlorinated compounds. This process could have started in the Late Silurian to Early Devonian, 400 million years ago, when the first soils on earth evolved. PMID- 12075811 TI - A model to predict long-term performance of vapor-phase bioreactors: a cellular automaton approach. AB - A novel numerical model was constructed to predict performance of vapor-phase bioreactors (VPBs) operated over extended periods. This model incorporates two unique features to simulate changes in pollutant removal efficiency and biomass accumulation: (1) total biomass is divided into two microbial components, active and inactive biomass, and (2) biomass growth and biofilm thickness changes are simulated by means of a cellular automaton (CA) approach. The CA approach, a differential-discrete algorithm, numerically allows the excess quantity of biomass in each numerical element to move toward the biofilm surface as biomass accumulates. One set of experimental bioreactor data was used to estimate unknown model parameters. A 90-day simulation using the estimated parameters agreed with pollutant removal and biomass accumulation profiles determined experimentally. Four additional model simulations using the same estimated model parameters were generally consistent with experimental data collected from a series of toluene degrading VPBs operated over a range of conditions. Model predictions imply that the decline in bioreactor performance observed over extended operation was caused by a decline in the active biomass fraction and a decrease in the biofilm specific surface area. This CA model provides insight into biomass accumulation during complex bioreactor operation and improves our capability to predict long term VPB performance. PMID- 12075809 TI - Resuspension of soil as a source of airborne lead near industrial facilities and highways. AB - Geologic materials are an important source of airborne particulate matter less than 10 microm aerodynamic diameter (PM10), but the contribution of contaminated soil to concentrations of Pb and other trace elements in air has not been documented. To examine the potential significance of this mechanism, surface soil samples with a range of bulk soil Pb concentrations were obtained near five industrial facilities and along roadsides and were resuspended in a specially designed laboratory chamber. The concentration of Pb and other trace elements was measured in the bulk soil, in soil size fractions, and in PM10 generated during resuspension of soils and fractions. Average yields of PM10 from dry soils ranged from 0.169 to 0.869 mg of PM10/g of soil. Yields declined approximately linearly with increasing geometric mean particle size of the bulk soil. The resulting PM10 had average Pb concentrations as high as 2283 mg/kg for samples from a secondary Pb smelter. Pb was enriched in PM10 by 5.36-88.7 times as compared with uncontaminated California soils. Total production of PM10 bound Pb from the soil samples varied between 0.012 and 1.2 mg of Pb/kg of bulk soil. During a relatively large erosion event, a contaminated site might contribute approximately 300 ng/m3 of PM10-bound Pb to air. Contribution of soil from contaminated sites to airborne element balances thus deserves consideration when constructing receptor models for source apportionment or attempting to control airborne Pb emissions. PMID- 12075810 TI - Theoretical method for lumping multicomponent secondary organic aerosol mixtures. AB - Atmospheric organic aerosol mixtures are composed of hundreds of individual semivolatile organic components, each with its own partitioning properties. In most atmospheric models, these components are grouped together into a few lumped compounds. A theoretical method is described for converting multiple organic aerosol components into one or more lumped groups using a set of equations that calculates the properties of lumped compounds from individual component properties. Expected organic aerosol mass and temperature range values are specified and components are grouped together according to relative volatility. A universal set of volatility dividing lines is presented for dividing components into multiple lumped groups. The lumping method is evaluated with three different mixtures: a base case mixture, a set of 1000 random mixtures with varying properties, and a mixture of products formed from the reaction of alpha-pinene and ozone. Modeling results suggest that lumping a multicomponent mixture into two or three groups may be sufficient to represent partitioning behavior over a wide range of temperature and organic aerosol mass. The temperature dependence of lumped groups is predicted to be lower than that of their individual components. PMID- 12075812 TI - Reactive transport in porous media: a comparison of model prediction with laboratory visualization. AB - Groundwater transport models that accurately describe spreading of nonreactive solutes in an aquifer can poorly predict concentrations of reactive solutes. The dispersive term in the advection-dispersion equation can overpredict pore-scale mixing, and thereby overpredict homogeneous chemical reaction. We quantified this experimentally by imaging instantaneous colorimetric reactions between solutions of aqueous CuSO4 and EDTA4- within a 30-cm long translucent chamber packed with cryolite sand that closely matched the optical index of refraction of water. A charge-coupled device camera was used to quantify concentrations of blue CuEDTA2- within the chamber as it was produced by mixing of the two reactants at different flow rates. We compared these experimental results with a new analytic solution for instantaneous bimolecular reaction coupled with advection and dispersion of the product and reactants. For all flow rates, the concentrations of CuEDTA2- recorded in the experiments were about 20% less than predicted by the analytic solution, thereby demonstrating that models assuming complete mixing at the pore scale can overpredict reaction during transport. PMID- 12075813 TI - Porous polysilsesquioxanes for the adsorption of phenols. AB - Arylene- and ethylene-bridged polysilsesquioxane materials have been synthesized by the hydrolysis and condensation of alkoxysilyl precursors under basic conditions. Cetyltrimethylammonium chloride was used to increase the porosity and surface areas of these materials via the surfactanttemplate approach. Structural characterization of these materials was carried out by nitrogen gas sorption and X-ray diffraction. The adsorption of three phenolic compounds (4-nitrophenol, 4 chlorophenol, 4-methylphenol) has been investigated by both batch and column testing. The arylene-bridged material exhibited a much greater affinity for all three phenols. The efficient removal of adsorbed phenols by a simple ethanol wash led to sorbent regeneration and separation of the aromatic species. PMID- 12075814 TI - UV disinfection of Giardia lamblia cysts in water. AB - The human and animal pathogen Giardia lamblia is a waterborne risk to public health because the cysts are ubiquitous and persistent in water and wastewater, not completely removed by physical-chemical treatment processes, and relatively resistant to chemical disinfection. Given the recently recognized efficacy of UV irradiation against Cryptosporidium parvum oocysts, the inactivation of G. lamblia cysts in buffered saline water at pH 7.3 and room temperature by near monochromatic (254 nm) UV irradiation from low-pressure mercury vapor lamps was determined using a "collimated beam" exposure system. Reduction of G. lamblia infectivity for gerbils was very rapid and extensive, reaching a detection limit of >4 log within a dose of 10 JM-2. The ability of UV-irradiated G. lamblia cysts to repair UV-induced damage following typical drinking water and wastewater doses of 160 and 400 JM(-2) was also investigated using experimental protocols typical for bacterial and eucaryotic DNA repair under both light and dark conditions. The infectivity reduction of G. lamblia cysts at these UV doses remained unchanged after exposure to repair conditions. Therefore, no phenotypic evidence of either light or dark repair of DNA damage caused by LP UV irradiation of cysts was observed at the UV doses tested. We conclude that UV disinfection at practical doses achieves appreciable (much greater than 4 log) inactivation of G. lamblia cysts in water with no evidence of DNA repair leading to infectivity reactivation. PMID- 12075815 TI - Beautifying wastewater treatment. PMID- 12075816 TI - Task-specific ionic liquids incorporating novel cations for the coordination and extraction of Hg2+ and Cd2+: synthesis, characterization, and extraction studies. AB - A series of hydrophobic task-specific ionic liquids designed to extract Hg2+ and Cd2+ from water were prepared by appending urea-, thiourea-, and thioether substituted alkyl groups to imidazoles and combining the resulting cationic species with PF6-. The new ionic liquids were characterized and investigated for their metal ion extraction capabilities. When used in liquid/liquid extraction of Hg2+ and Cd2+ from aqueous solutions, the metal ion distribution ratios increased several orders of magnitude, regardless of whether the ionic liquids were used as the sole extracting phase or doped into a series of [1-alkyl-3 methylimidazolium][PF6] (alkyl = n-C4-C8) ionic liquids to form a 1:1 solution. In the 1:1 mixtures, as the length of the alkyl chain increased from butyl to hexyl to octyl, the metal ion distribution ratios increased. Increasing the ratio TSIL/[C4mim][PF6] resulted in higher distribution ratios for both Hg2+ and Cd2+. Overall, the thiourea- and urea-derivatized cations yielded the highest distribution ratios, and those for Hg2+ were higher than those for Cd2+; however, a change in aqueous-phase pH does not promote the stripping of metal ions from the extracting phase. The combination of these imidazolium cations and PF6- produced ionic liquids with decreased thermal stability in comparison to [C(n)mim]-[PF6]. Gaussian98 restricted Hartree-Fock geometry optimizations for one of the thiourea-appended cations shows the charge delocalization around the ring and suggests that the thiourea group may aid in deprotonating the imidazolium ring and may be responsible for the lowered thermal stability of these cations. PMID- 12075817 TI - Biological hydrogen production measured in batch anaerobic respirometers. AB - The biological production of hydrogen from the fermentation of different substrates was examined in batch tests using heat-shocked mixed cultures with two techniques: an intermittent pressure release method (Owen method) and a continuous gas release method using a bubble measurement device (respirometric method). Under otherwise identical conditions, the respirometric method resulted in the production of 43% more hydrogen gas from glucose than the Owen method. The lower conversion of glucose to hydrogen using the Owen protocol may have been produced by repression of hydrogenase activity from high partial pressures in the gastight bottles, but this could not be proven using a thermodynamic/rate inhibition analysis. In the respirometric method, total pressure in the headspace never exceeded ambient pressure, and hydrogen typically composed as much as 62% of the headspace gas. High conversion efficiencies were consistently obtained with heat-shocked soils taken at different times and those stored for up to a month. Hydrogen gas composition was consistently in the range of 60-64% for glucose-grown cultures during logarithmic growth but declined in stationary cultures. Overall, hydrogen conversion efficiencies for glucose cultures were 23% based on the assumption of a maximum of 4 mol of hydrogen/ mol of glucose. Hydrogen conversion efficiencies were similar for sucrose (23%) and somewhat lower for molasses (15%) but were much lower for lactate (0.50%) and cellulose (0.075%). PMID- 12075818 TI - Poly[N-(2-hydroxypropyl)methacrylamide] conjugates of bovine pancreatic ribonuclease (RNase A) inhibit growth of human melanoma in nude mice. AB - Recently hydrophilic poly[N-(2-hydroxypropyl)methacrylamide] (PHPMA) was used for BS-RNase modification to prevent its degradation in bloodstream or fast elimination. Polymer-conjugated BS-RNase preparations proved to be cytotoxic after intravenous or intraperitoneal application, whereas native BS-RNase was ineffective. Here RNase A unimer was conjugated with two HPMA polymers (classic and star) and their antitumor effects both in vitro and in vivo were compared with those of BS-RNase polymers. Surprisingly, the antitumor effect of RNase A conjugates was also pronounced. The RNase A conjugates (classic and star) injected intravenously to mice bearing melanoma tumor caused a significant reduction in tumor volume following ten doses of 5 and 1 mg/kg, respectively. Despite the antitumor activity observed in vivo, the in vitro tested cytotoxic activity of RNase A did not differ from that caused by native RNase A while native BS-RNase (50 microg/ml) totally inhibited DNA synthesis in treated cells. The experiments with 125I-labeled preparations demonstrated concentration dependent internalization of native BS-RNase by tumor cells within an hour, whereas the polymer conjugate (S-BS) was not internalized. On the contrary, the in vivo experiments showed that whereas 40% of S-BS conjugate persisted in bloodstream for 24h after administration, 98% of the native BS-RNase was already eliminated. Improved antitumor activities of PHPMA-modified RNases in vivo might be ascribed to their prolonged retention in bloodstream, better proteolytic stability and resistance to the action of the ribonuclease inhibitor. PMID- 12075819 TI - Immunomodulator tuftsin augments anti-fungal activity of amphotericin B against experimental murine candidiasis. AB - In the present study, we report the potential of an immunomodulator tuftsin in increasing the efficacy of liposomised Amphotericin B (Amp B) against drug sensitive as well as drug resistant experimental murine candidiasis. The Amp B containing liposomes demonstrated strong potential of eliminating systemic candidiasis (70% survival) in animals infected with Amp B sensitive strain of Candida albicans (C. albicans). The same liposomal formulation was found to be ineffective in treatment of animals infected with drug resistant C. albicans. However, the co-administration of liposomal formulation of Amp B along with an immunomodulator tuftsin, was found to be competent enough in curing even the drug resistant candidiasis. In contrast, none of the animals survived in the control groups, which were treated with free or liposomised Amp B (without tuftsin). Further, the effect of liposomised tuftsin, on T-cell proliferation as well as antibody production reveals that tuftsin elicits strong immunopotentiating effects as well. The pretreatment with liposomised tuftsin prior to challenging the animals with drug resistant C. albicans infection has also been effective and shows an extra edge in prophylactic perspectives. PMID- 12075820 TI - Ultrasound enhancement of in vitro transfection of plasmid DNA by a cationized gelatin. AB - In vitro transfection efficiency of a plasmid DNA for rat gastric mucosal (RGM)-1 cells was enhanced by ultrasound (US) irradiation. Ethylenediamine was introduced to the carboxyl groups of gelatin to prepare a cationized gelatin as the vector of plasmid DNA encoding luciferase. An electrophoresis experiment revealed that the cationized gelatin was mixed with plasmid DNA at the weight ratio of 5.0 to form a cationized gelatin-plasmid DNA complex. The complex obtained was about 200nm in diameter with a positive charge. When incubated with the cationized gelatin-plasmid DNA complex and subsequently exposed to US, RGM-1 cells exhibited a significantly enhanced luciferase activity although the extent increased with an increase in the DNA concentration, in contrast to the cationized gelatin alone with or without US irradiation and US irradiation alone. US irradiation was also effective in enhancing the activity by free plasmid DNA although the extent was less than that of the complex. The US-induced enhancement of luciferase activity was influenced by the exposure time period, frequency, and intensity of US. The activity enhancement became higher to be significant at the irradiation time period of 60 s and thereafter decreased. A series of cytotoxicity experiments revealed that an increase in the irradiation time period and intensity of US decreased the viability of cells themselves. It is possible that US irradiation under an appropriate condition enables cells to accelerate the permeation of the cationized gelatin-plasmid DNA complex through the cell membrane, resulted in enhanced transfection efficiency of plasmid DNA. These findings clearly indicate that US exposure is a simple and promising method to enhance the gene expression of plasmid DNA. PMID- 12075821 TI - Ultrasound exposure enhances the biological action of interferon in the liver. AB - Although interferon (IFN) alpha and beta are currently recognized as the most effective agents for treating patients with chronic hepatitis B and C, they are well known to cause various adverse effects. To reduce the dose of IFN necessary for treatment, we tried enhancing the effects of IFN in the liver by ultrasound exposure. Percutaneous insonation in mouse liver following IFN-beta injection with the ultrasound power level used at clinical diagnosis enhanced the IFN-beta induced increase of 2',5'-oligoadenylate synthetase (2-5AS) levels in the liver. This enhancement of the 2-5AS level was dependent on the duration as well as on the timing of insonation after the IFN-beta injection. In contrast, liver insonation did not enhance 2-5AS levels in the lung or spleen, and moreover, it did not alter tissue distribution of injected IFN. Thus, the combination of IFN beta administration and subsequent liver insonation appears to be a promising method for enhancing the antiviral activity of IFN specifically in the liver, enabling a reduction in the dose necessary for treatment. PMID- 12075822 TI - Polyester microparticles as a vaccine delivery system for brucellosis: influence of the polymer on release, phagocytosis and toxicity. AB - Microparticles, containing an antigenic complex from Brucella ovis (HS), were evaluated for vaccine purposes against brucellosis. They were prepared by the double emulsion solvent evaporation method using two different polyesters, poly lactide-co-glycolide acid (75:25; RG 756) and poly-epsilon-caprolactone. The encapsulation efficiency and release of HS from the microparticles, their capacity to be phagocytosed and also their toxicity on murine monocytes J774.2 were investigated. Both polymers lead to smooth and spherical sub-5 microm particles, with approximately 30% of the antigen initial dose encapsulated. SDS PAGE and immunoblot of extracted antigens confirmed that the apparent molecular weight and antigenicity remained unaltered after the encapsulation procedure. However, the in vitro release of the antigens differed among them. The release profile for PLGA microparticles was continuous, whereas PEC ones released the antigens in a triphasic release pattern. Phagocytosis was clearly influenced by the hydrophobicity of the polymer, increasing in the case of PEC microparticles. Toxicity assay showed that both types of microparticles induced similar levels of mitochondrial damage. In conclusion, HS-PEC microparticles could be used as an effective vaccine against brucellosis, as the antigen is released in boosters and they are greatly phagocytosed by macrophages. PMID- 12075823 TI - A novel formulation for superparamagnetic iron oxide (SPIO) particles enhancing MR lymphography: comparison of physicochemical properties and the in vivo behaviour. AB - The major aim of this study was to prove or disprove the theories concerning the correlation between physicochemical properties of superparamagnetic iron oxide (SPIO) particles and their accumulation in the lymph nodes. New SPIO particles were produced using starch as stabilising polymer shell. The synthesis was done in a two-step procedure using conventional wet-chemical precipitation technique and subsequent coating of the iron oxide cores. The particles were physicochemically characterised and their lymphotrophy studied in rats using well described lymphotropic dextran-coated SPIO particles as reference. Despite the short blood half-lives of approximately 13 min and the relatively large sizes (approximately 60-90 nm), the starch SPIO particles proved at least as efficient in lymph node accumulation as the small 25 nm dextran SPIO particles having a half-life of 90 min. The currently accepted theories concerning the connection between particle properties and their uptake into lymph nodes are not generally valid, or have at least to be limited for dextran-coated SPIO particles. Lymph node targeting could be achieved despite the present theories consider small size (<30 nm) and long circulation times in the blood as prerequisites. Histological examination showed, that SPIO particles could only be found in lymph node areas where macrophages could be marked which enclosed the particles. Localisation in marginal areas of the lymph nodes indicates endothelial transcytosis as the major accumulation pathway. PMID- 12075824 TI - Efficacy of ganciclovir-loaded nanoparticles in human cytomegalovirus (HCMV) infected cells. AB - The use of albumin nanoparticles to enhance the antiviral activity of ganciclovir (GCV) while decreasing its intrinsic toxicity was evaluated in human fibroblasts. GCV was adsorbed onto preformed protein nanoparticles (Np A) or incubated with the albumin solution prior to the formation of nanoparticles (Np B) by a coacervation method. The antiviral efficacies in MRC-5 and CHN cells were assayed by plaque reduction assay and early antigen detection, with several MOI and time of drug addition (T0 and T48). Whatever cell line or assay tested, Np A is the most active formulation whereas the efficacy of Np B is similar to the ganciclovir conventional therapy. Moreover, the profile of the dose-activity curve of the drug as a function of MOI is not altered by the use of nanoparticles and the efficacy of all formulations improves when added at T48. On the other hand, Np B produces a decrease on the cytotoxicity of the free drug in non infected cells. Both activity and cytotoxicity seem to be straight correlated to the drug internalisation by cells. Thus, Np A highly improves the drug uptake, whereas Np B leads to a similar drug internalisation than the free drug. PMID- 12075825 TI - Pharmacokinetics and brain uptake of biotinylated basic fibroblast growth factor conjugated to a blood-brain barrier drug delivery system. AB - Human basic fibroblast growth factor (bFGF) is a potent neuroprotective agent. The clinical efficacy of this neurotrophin, however, is restricted by poor permeability across the blood-brain barrier (BBB). This study was designed to test the hypotheses that bFGF will retain its biological activity and have an enhanced BBB transport after re-formulation and conjugation to a BBB peptide drug delivery vector. The BBB delivery vector is comprised of a conjugate of streptavidin (SA) and the murine OX26 monoclonal antibody against the rat transferrin receptor, and the conjugate of biotinylated bFGF (bio-bFGF) bound to a vector is designated bio-bFGF/OX26-SA. A radioreceptor binding assay shows that the native bFGF, bio-bFGF, and bio-bFGF/OX26-SA conjugate have IC50 values of 0.12, 0.40, and 0.56 nM, respectively. After an IV bolus injection to the rat, [125I]-bio-bFGF is avidly taken up by peripheral organs, with low brain uptake at 60 min, 0.010+/-0.004% of injected dose (ID)/g brain. By contrast, the brain uptake of the [125I]-bio-bFGF/OX26-SA is increased 5-fold to 0.050+/-0.011%ID/g, although the uptake of the conjugate by peripheral tissues was decreased relative to the unconjugated bio-bFGF. In conclusion, conjugation of bio-bFGF to a BBB drug delivery vector (a) causes only a minor decrease in affinity for the bFGF receptor, (b) decreases the peripheral organ uptake of the bFGF, and (c) increases the brain uptake of the neurotrophin. The re-formulation of bFGF to enable receptor-mediated transcytosis across the BBB may improve the therapeutic index of this neurotrophin as a neuroprotective agent. PMID- 12075826 TI - Studies on the development of colon-targeted delivery systems for celecoxib in the prevention of colorectal cancer. AB - The nonsteroidal anti-inflammatory drugs (NSAIDs) are found to be potential chemopreventive agents of colorectal cancer. Celecoxib, an NSAID with selective cyclooxygenase-2 inhibition, was proved to be effective for the prevention of colon cancer in patients with familial adenomatous polyposis (FAP) and sporadic polyps. In the light of this information, the present study was carried out to develop oral colon-targeting drug delivery systems for celecoxib using guar gum as a carrier. Matrix tablets containing various proportions of guar gum were prepared by wet granulation technique using starch paste as a binder. The tablets were evaluated for hardness, drug content and were subjected to in vitro drug release studies. The amount of celecoxib released from the matrix tablets at different time intervals was estimated by a HPLC method. Guar gum matrix tablets released only 2-4% of celecoxib in the physiological environment of stomach and small intestine depending on the proportion of guar gum used in the formulation. When the dissolution study was continued in simulated colonic fluids (rat caecal content medium), the matrix tablets containing 20% of guar gum released another 37% of celecoxib after degradation by the colonic bacterial action. The matrix tablets containing 30% of guar gum released about 24% of celecoxib in simulated colonic fluids indicating the susceptibility of the guar gum formulations to the rat caecal contents. The results of the study show that the matrix tablets containing either 20 or 30% of guar gum are most likely to target celecoxib for local action in the colon. The guar gum matrix tablets of celecoxib showed no change either in physical appearance, drug content or in dissolution pattern after storage at 40 degrees C/RH 75% for 6 months. Differential scanning calorimetry (DSC) studies indicated no possibility of interaction between celecoxib and guar gum/other formulation excipients. PMID- 12075828 TI - Abdominal compartment syndrome. PMID- 12075827 TI - Absorption enhancing effect of labrasol on the intestinal absorption of insulin in rats. AB - The oral absorption enhancing effect of Labrasol has been studied in rats using insulin as a model peptide/protein drug. Insulin solution was prepared by dissolving insulin in pH 7.4 buffer followed by the addition of Labrasol. The insulin concentration was 50.0 IU/ml. The test insulin/Labrasol solution was administered to the jejunum, ileum and ascending colon of rats at 10.0 IU/kg. After administration, blood samples were collected for 5 h and serum glucose levels and insulin levels were measured. In another group of rats, insulin solution was injected intravenously at 1.0 IU/kg, and both serum glucose and insulin levels were measured. The pharmacological availability of insulin from Labrasol solution was found to be 3.9, 8.9 and 9.1% following jejunal, ileal and colonic administrations, respectively, by comparing the serum glucose level vs. time profiles obtained after intestinal and i.v. administrations. By comparing the serum insulin levels vs. time profiles, the bioavailability of insulin was found to be 0.25 and 0.20% for intra-ileum and colonic administrations, respectively. The hypoglycemic effect of insulin after intra-ileum administration showed a dose-dependency in the insulin dose range from 10.0 to 1.0 IU/kg. These results suggest the absorption enhancing effect of Labrasol on the intestinal absorption of insulin in rats. PMID- 12075829 TI - Vascular trauma: a 40-year experience with extremity vascular emphasis. PMID- 12075830 TI - Trauma scoring. AB - Severity scales to characterize the nature and extent of injury are important adjuncts to trauma care systems, trauma research and many of the elements of a complete public health approach to injury. This article provides a brief overview of severity scale development over the past 30 years during which the science to support such initiatives has matured substantially. Anatomical, physiological, intensive care, composite and complex scales and models now abound and are being applied to a variety of tasks with increasing precision. Trauma registries enable the meaningful aggregation of data for the development and testing of models. Future challenges are identified as are potentially fruitful avenues of research. PMID- 12075831 TI - Initial management of the poly-trauma patient: a practical approach in an Australian major trauma service. AB - The initial management of the poly-trauma patient is of vital importance to minimizing both patient morbidity and mortality. We present a practical approach to the early management of a severely injured patient as practiced at Liverpool Hospital in Sydney, Australia. Specific attention is paid to innovations in care and specific controversies in early management as well as local solutions to challenging problems. PMID- 12075832 TI - Penetrating neck injuries. PMID- 12075833 TI - Penetrating injuries of the chest: indications for operation. AB - Penetrating injuries to the chest present a frequent and challenging problem. The majority of these injuries can be managed non-operatively. The selection of patients for operation or observation can be made by clinical examination and appropriate investigations. The trauma ultrasound has become a valuable first line tool to rule out pericardial tamponade. Spiral computed tomography of the chest is increasingly used to evaluate transmediastinal gunshot wounds and direct, if needed, further organ-specific tests, such as esophagography, aortography, or bronchoscopy. Minimally invasive techniques have found sound application in the thoracoscopic evacuation of undrained hemothorax and the laparoscopic evaluation of diaphragmatic trauma. In the operative arena, lung sparing techniques with the use of staplers, like wedge resection and tractotomy, have allowed easier, faster, and effective control of bleeding without sacrificing unnecessarily normal pulmonary parenchyma. Knowledge of the new advancements in the field of thoracic trauma will allow surgeons to provide expert care and improved outcomes. PMID- 12075834 TI - Advances in the management of thoracic vascular injury. PMID- 12075835 TI - Trauma surgery. PMID- 12075836 TI - Blunt abdominal trauma: evaluation and indications for laparotomy. PMID- 12075837 TI - When to operate on abdominal stab wounds. PMID- 12075838 TI - When to operate on abdominal gunshot wounds. AB - Historically, penetrating abdominal trauma was managed expectantly until the late 19th century. In World War I, with the high mortality and morbidity associated with penetrating abdominal trauma, operative management replaced expectant management. It was soon realized that not all penetrating abdominal injuries required an operation. Since the 1960's, selective nonoperative management of stab wounds to the anterior abdomen has become the standard of care. However, gunshot wounds to the abdomen are still treated by mandatory exploration based on an allegedly high incidence of intra-abdominal injuries and low rate of complications, if laparotomy turns out negative. A number of series have recently surfaced, reporting successful outcomes, while decreasing morbidity and hospital length of stay, with selective non-operative management of gunshot wounds to the abdomen. This review will address the current controversies surrounding selective nonoperative management of gunshot wounds to the abdomen and will present our experience and current approaches. PMID- 12075839 TI - Cavitary endoscopy in trauma: 2001. AB - Laparoscopy and thoracoscopy have been used in the evaluation of injured patients for over 30 years. Despite this long history, indications for use of these techniques remains controversial. The widespread availability of videoscopic equipment which followed the introduction of laparoscopic cholecystectomy increased interest and utilization of minimally invasive techniques in evaluation of trauma patients. Laparoscopy has been most beneficial in the evaluation of hemodynamically stable victims of stabbings and gunshots. This technique has primarily been used to detect peritoneal penetration in tangential wounds of the abdominal wall and for evaluation of the diaphragm in patients with thoracoabdominal wounds. Laparoscopic evaluation in blunt trauma patients is of unproven utility, but has been used in the assessment of patients with documented solid organ injury and in the evaluation of patients with suspected hollow viscus injury. Small subsets of patients are candidates for therapeutic laparoscopic interventions, i.e., suture repair of diaphragmatic lacerations. Thoracoscopy or videoassisted thoracic surgery (VATS) is useful for evaluation of the diaphragm, early evacuation of clotted hemothorax, and assessment of ongoing bleeding. PMID- 12075840 TI - Organization of trauma care. PMID- 12075841 TI - Hepatic trauma. PMID- 12075842 TI - Pancreatic and duodenal injuries. complex and lethal. PMID- 12075843 TI - Surgical options in colorectal injuries. AB - Colonic or rectal injuries occur in up to 10% of patients that suffer penetrating or severe blunt abdominal trauma. The majority of colon injuries are diagnosed intraoperatively following a penetrating abdominal injury. Rectal injuries are usually diagnosed preoperatively with a high index of suspicion based upon the wounding missile trajectory. The vast majority of colon injuries can be primarily repaired with a significant trend toward avoiding colostomy whenever possible. Colostomy is increasingly reserved for rectal injuries and destructive colon injuries with extenuating circumstances such as hemodynamic instability and significant associated injuries. PMID- 12075844 TI - Damage control surgery. PMID- 12075845 TI - Determining the impact of health library services on patient care: a review of the literature. AB - The impact of health library information has been studied in a number of settings and populations. This review examines both the methods employed and the outcomes reported in a series of impact studies conducted in four countries since the late 1980's. It demonstrates that health library information can affect patient care outcomes in various ways and has high cognitive and clinical value in diagnosing and treating patients. But there remains no agreed technique or methodology for determining the level of this impact. In many cases evaluation of the health library's impact will be located within a central process of evaluation of the greater organization to which the library belongs. Consequently, local ad hoc evaluation is rarely reported outside an organization. The author considers the question of whether it is worthwhile doing an impact study and what form it might take. The author concludes by identifying seminal papers on the topic. PMID- 12075846 TI - Improving access to the international coverage of reports of controlled trials in electronic databases: a search of the Australasian Medical Index. AB - INTRODUCTION: Locating reports of trials from journals not indexed in the major databases presents difficulties to systematic reviewers, and may be a factor in improving the reliability of the reviews. OBJECTIVES: To identify and make available reports of controlled trials from the Australasian Medical Index (AMI). To measure the quality of indexing of trials in AMI. METHODS: Using a highly sensitive search strategy consisting of methodology indexing and free-text terms, records from AMI were read for reports of controlled trials. Trials meeting the criteria were submitted for inclusion in The Cochrane Controlled Trials Register (CCTR) and assessed for the quality of their indexing. RESULTS: 3621 records were downloaded, of which 512 were identified as reports of controlled trials (317 RCTs; 195 CCTs) and submitted to CCTR. The precision of methodology indexing terms was 60%, but sensitivity just 18%. The quality of indexing of trials was generally poor with only 50 tagged with the RCT/CCT publication type term. 453 reports (88%) were not previously available in CCTR. CONCLUSIONS: The large proportion of trials found to be unique to the AMI database increases the pool of studies available to systematic reviewers, and helps ensure CCTR remains the most comprehensive source of trials. PMID- 12075847 TI - The ASVIN Project: a study of electronic and printed veterinary science journals in the UK and Ireland. AB - ASVIN was a Research Support for Libraries Programme (RSLP) Funded Project, whose principal aim was to develop better co-operation between nine libraries in the veterinary and animal health field. The project examined a number of library related areas, such as journal archiving, collection development policies, development of a common Z39.50 catalogue interface, grey literature, and printed and electronic journals. This paper examines the latter, whilst also providing a detailed analysis of printed journals held in partner libraries. Comparisons of the nine partner library collections are undertaken with a range of secondary sources, and an ASVIN core list of veterinary journals was developed. It looks at some of the issues relating to e-journals, expressing the benefits and concerns that many librarians are experiencing, such as licensing arrangements, archiving and general management issues. PMID- 12075848 TI - Different for girls? Feminism, health information and librarianship. AB - This paper focuses on the provision and organization of health information materials in women's health centres in UK and Ireland in the late 20th century Such centres sprung from the work of feminist activists and health workers from the late 1960s onwards, promoting health information and other interventions to counteract women's devalued status within society, and the stereotypes perpetuated by health care and other systems. Centres that developed were (and still are) typically within the voluntary sector, have a strong feminist perspective and are run by lay workers. This paper will draw on research into information provision in these centres, its scope, organization and who provides it. It will argue that this work is of interest to mainstream librarianship, but there are minimal linkages as feminist thinking within librarianship has been unable overall to make common cause with the work of these, and other such agencies, which has inhibited potential developments of mutual benefit. This paper draws on ongoing research into feminism and librarianship, and findings that have been presented in a number of settings. PMID- 12075849 TI - Quality of web-based medical information on stable COPD: comparison of non commercial and commercial websites. AB - The Internet provides an easy and accessible way to deliver medical information about the management of various diseases, both to practitioners and to their patients. As there is no control over who posts information on the Web, there is a risk that the interests of the web producer may bias the quality of information. The quality of medical information on the management of chronic obstructive pulmonary disease (COPD) on the Internet was evaluated, comparing non commercial and commercial websites. An internet search was conducted to locate relevant websites using a metasearch engine. The quality of websites was scored on a scale of 0-10, based on three items about the credibility of the site and seven items about the accuracy of the information provided by the site. Quality differences between commercial and non-commercial websites were explored. The search revealed 23 relevant websites (12 noncommercial and 11 commercial). The overall quality of non-commercial websites was better than that of commercial websites (median score 7 vs. 4, P = 0.01). Compared to commercial sites, non commercial websites more often provided information about cessation of smoking (100% vs. 64%, P = 0.03), preventative influenza vaccinations (42% vs. 9%, P = 0.07) and use of long-term oxygen therapy (92% vs. 45%, P = 0.02). Among websites providing information on COPD, commercial sites were much more likely to be of poorer quality compared to sites of non-commercial organizations. In particular, commercial sites do not provide information about simple preventative treatments. There is a need to be vigilant about the quality of health information about COPD on the Internet. PMID- 12075850 TI - Local approaches to information for health. PMID- 12075851 TI - Clinical librarians at Barnet Primary Care NHS Trust: addressing the information requirements of clinical governance. PMID- 12075852 TI - Mirage or reality? PMID- 12075853 TI - The actions of a D-1 agonist in MPTP treated primates show dependence on both D-1 and D-2 receptor function and tolerance on repeated administration. AB - The potent and long acting D-1 receptor agonist, A-77636 reverses motor deficits in MPTP treated common marmosets following subcutaneous or oral administration. We now explore the effects of acute versus repeated administration of A-77636 and the relative roles of D-1 and D-2 receptor involvement in its antiparkinsonian actions. Acute oral administration (0.18-9.0 mg/kg) or subcutaneous administration (0.036-1.08 mg/kg) of A-77636 produced well co-ordinated bouts of discontinuous locomotor activity. There was no evidence of repetitive or stereotyped movements. However, oral administration of A-77636 (1.44 mg/kg) on alternate days for 10 days produced tolerance to the increase in locomotor activity and improvement of disability. Pretreatment with the D-1 antagonist SCH 23390 (0.625, 2.5 or 5.0 mg/kg, intraperitoneally), produced a dose dependent reduction in locomotor activity and antagonised the reduction in disability scores following administration of A-77636 (0.36 mg/kg, subcutaneously). The inhibitory effects of SCH 23390 lasted for some 2-5 hours after which the activity of A-77636 was gradually restored. Unexpectedly, pretreatment with the D 2 antagonist raclopride (1.25, 5.0 or 20.0 mg/kg, intraperitoneally), dramatically diminished the antiparkinsonian effects of A-77636 (0.36 mg/kg, subcutaneously) in a dose dependent manner. The dependence of the antiparkinsonian activity of A-77636 on intact D-2 receptor function, suggests a need for endogenous D-2 receptor tone to express D-1 mediated locomotor activity. PMID- 12075854 TI - An early methamphetamine challenge suppresses the maturation of dopamine fibres in the nucleus accumbens of gerbils: on the significance of rearing conditions. AB - The effect of a single early methamphetamine (MA) challenge on postnatal maturation of the nucleus accumbens (NAC) was studied. Therefore, male gerbils received a single dose of MA (50 mg/kg, i.p.) on postnatal day 14. At the age of postnatal day 90, dopamine fibres were stained immunocytochemically and innervation density was determined in several test fields along the rostrocaudal extent of both core and shell of the NAC. Since we already know that the differential environment can alter ontogeny of dopamine innervation in the prefrontal cortex of gerbils, in the present study we investigated whether probable drug effects may be influenced by rearing conditions. For that purpose, animals were bred and reared either isolated in standard laboratory cages or grouped in an object-filled environment. The results showed that a single early MA challenge significantly alters maturation of dopamine fibre innervation in both subregions of the NAC. In seminaturally reared gerbils the drug challenge caused dopamine fibre densities which were about 54% below those of saline treated controls in both the shell and core. However, in animals from restricted rearing this MA-induced effect was more pronounced in the core (-43%) but not significant in the shell (-14%). In conclusion, an early MA challenge caused a significant restraint of adult dopamine fibre density developing in the NAC postnatally. Additionally, rearing conditions significantly interfered with drug induced alterations in maturation of dopaminergic innervation pattern of the NAC. The present results are discussed with recent findings on MA-induced impairment of prefrontal dopamine innervation and further reactive morphogenetic effects caused by the drug. In this respect, functional interactions between the prefrontal cortex and NAC are specifically considered. PMID- 12075856 TI - Estrogen induces elevation of cAMP-dependent protein kinase activity in immortalized hippocampal neurons: imaging in living cells. AB - 17beta-estradiol-stimulated PKA activation in living differentiated H19-7 cells was visualized using DR2, a cell-permeable fluorescent PKA substrate. Estradiol induced marked changes of fluorescence in cytosolic areas and these were inhibited by a PKA inhibitor. PMID- 12075855 TI - Protection of dopaminergic neurons in primary culture by lisuride. AB - Dopamine agonists play an important role in the treatment of Parkinson's disease by reducing the administration of L-3,4-dihydroxyphenylalanine (L-DOPA). The enzymatic and non-enzymatic conversion of L-DOPA is suspected to increase oxidative stress, which leads to the degeneration of dopaminergic neurons in Parkinson's disease. In primary mouse mesencephalic cultures we show that the dopamine D1/D2 receptor agonist lisuride, in a concentration range of 0.001-1 microM, enhances the survival of dopaminergic neurons, protects against toxicity induced by L-DOPA or 1-methyl-4-phenylpyridinium ion (MPP+) and stimulates 3H dopamine uptake. Lisuride also reduces anaerobic metabolism during incubation with L-DOPA. The present findings suggest that lisuride may have trophic/survival promoting properties and potentially reduces oxidative stress. PMID- 12075857 TI - 3-OMD and homocysteine plasma levels in parkinsonian patients. AB - One main metabolizing pathway of levodopa is O-methylation to 3-O-methyldopa (3 OMD) by catechol-O-methyltransferase (COMT). Since COMT requires Mg2+ and S adenosylmethionine as methyl donor for this transmethylating process, COMT converts S-adenosylmethionine to S-adenosylhomocysteine and subsequent homocysteine. Objective of this study was to demonstrate relations between plasma levodopa, 3-OMD and total homocysteine in treated parkinsonian subjects. We measured homocysteine, levodopa and 3-OMD by HPLC. We compared plasma homocysteine in two groups of treated parkinsonian subjects subdivided according to their 3-OMD level. Homocysteine was significantly (p = 0.002) elevated in the group with higher 3-OMD concentrations and positively (r = 0.52, p = 0.0006) correlated to 3-OMD. Homocysteine induces vascular disease. Previous studies showed an increase of ischaemic heart- and cerebrovascular disease in treated parkinsonian patients. PMID- 12075858 TI - Increased neopterin production and tryptophan degradation in advanced Parkinson's disease. AB - Large amounts of neopterin are produced by interferon-(IFN)-gamma-stimulated human monocytes/macrophages, and increased neopterin concentrations indicate cellular immune activation. In parallel, IFN-gamma induces indoleamine 2,3 dioxygenase which degrades 1-tryptophan to kynurenine. Increased tryptophan degradation rates are indicated by an increased kynurenine/tryptophan ratio (kyn/trp-ratio), reflecting immune system activation, too. In 22 patients with Parkinson's disease (PD) and in 11 age-matched controls, serum and cerebrospinal fluid (CSF) neopterin concentrations were measured by ELISA. Tryptophan and kynurenine concentrations were determined by HPLC. Neopterin concentrations and kyn/trp-ratios were increased both in serum and CSF of patients as compared to controls. Serum tryptophan was lower in patients. Patients with the highest disease activity presented with highest degree of immune activation. Significant correlations existed between neopterin concentrations and kyn/trp-ratios in serum and CSF. Increased formation of neopterin and enhanced degradation of tryptophan suggest activated cell-mediated immune response in a subgroup of patients with advanced Parkinson's disease. PMID- 12075859 TI - Substantia nigra echogenicity is normal in non-extrapyramidal cerebral disorders but increased in Parkinson's disease. AB - Transcranial sonography (TCS) revealed substantia nigra (SN) hyperechogenicity in idiopathic Parkinson's disease (IPD). To further evaluate specificity of this finding, we examined 30 IPD patients and 30 age-matched subjects with non extrapyramidal cerebral disorders (NED). All IPD patients showed a SN hyperechogenicity, in 17 it was bilateral and in 13 unilateral. 7 NED patients had a SN hyperechogenicity, in all it was unilateral, confirming previous results in healthy subjects. Bilateral SN hyperechogenicity indicates IPD and normal SN echogenicity indicates NED. In 30% of patients TCS does not distinguish between IPD and NED. Data further support the assumption that bilateral SN hyperechogenicity is specific for IPD. PMID- 12075860 TI - Apraxia of eyelid closure in Huntington's disease. AB - We report a patient with genetically confirmed Huntington's disease (HD) presenting apraxia of eyelid closure (AEC). She was unable to close her eyes at command but was able to blink. Chorea and AEC ameliorated significantly during treatment with olanzapine and riluzole, an inhibitor of glutamate release. AEC is reported in progressive supranuclear palsy, Creutzfeldt-Jakob's disease, amyotrophic lateral sclerosis, and as post-stroke AEC. No report on HD is available so far, although oculomotor disturbances are quite common in this disease. PMID- 12075861 TI - Catatonic schizophrenia has a shorter pre-hospitalisation interval than other types of schizophrenia. AB - The time elapsed between the first onset of symptoms and the onset of first hospitalisation was analysed in 355 participants diagnosed with paranoid, simple, hebephrenic and catatonic schizophrenia and shizoaffective disorder. The real onset of the disease was assessed from interviews with reliable relatives and by reviewing medical records in general practices and out-patient psychiatric services. In 184 patients a family history of schizophrenia was identified. A positive family history was found to significantly increase the interval preceding first hospitalisation in all analysed types, except in catatonic schizophrenia. Possible reasons causing this prolongation are discussed, as well as repercussions of the results to studies using age of first hospitalisation as the leading indicator. PMID- 12075862 TI - Association study of C825T polymorphism of the G-protein b3 subunit gene with schizophrenia and mood disorders. AB - Alterations of G proteins have been implicated in major psychiatric illnesses. A C825T polymorphism of a gene encoding the beta3 subunit of heterotrimeric G proteins (GNB3) was reported to be associated with several pathological conditions, such as hypertension and depressive disorder. We examined whether this polymorphism is associated with functional psychoses in a Japanese sample of 370 schizophrenics, 164 bipolars, 68 depressive patients, and 198 controls. We obtained no evidence for an association of the polymorphism with any diagnostic group. PMID- 12075863 TI - Structural brain abnormalities specific to childhood-onset schizophrenia identified by neuroimaging techniques. AB - This review discusses functional and structural brain abnormalities in childhood onset schizophrenia identified by neuroimaging techniques. Published literature regarding both morphological and functional neuroimaging is discussed, regarding also the diversity of neuroimaging findings which partly reduces their reliability. The findings in early onset schizophrenia are compared with those of adult patients. The results of long-term investigations of structural abnormalities in early onset schizophrenia are given particular attention. The most consistent findings are ventricular enlargement and reduced total brain volume. Further, volumetric changes in the temporal and frontal cortex, thalamus, basal ganglia and limbic system are reported, as are hemispheric asymmetries and, conversely, reduction of normal differences. Findings regarding the corpus callosum and cerebellum are less consistent. In patients whose schizophrenia commenced in early childhood, the differences were generally more marked than in adolescence- or adult-onset schizophrenia. Atrophy of total brain volume was progressive throughout the course of the disorder. It is probable that neuroanatomical cerebral abnormalities present prior to disease onset play an etiopathogenic role in the development of schizophrenia. PMID- 12075864 TI - Determination of the target ventricular rate in patients with atrial fibrillation by evaluation of ventriculoarterial coupling. AB - In a pilot study, we determined the target ventricular rate of patients with atrial fibrillation by evaluating their ventriculoarterial coupling. Eleven patients with atrial fibrillation were studied. We recorded M-mode echocardiograms and radial artery blood pressure simultaneously. The left ventricular end-systolic pressure-volume ratio (Emax) and effective arterial elastance (Ea) were calculated for each beat, and the relationship of the preceding R-R interval (pRR) to Emax and Ea was evaluated. There was a significant positive correlation between pRR and Emax, and a significant negative correlation between pRR and Ea in all patients. The pRR that produced maximal stroke work was determined at the point of Emax=Ea, and the pRR that achieved maximal mechanical efficiency was determined at the point of 2Ea=Emax. By evaluating ventriculoarterial coupling in these patients who had atrial fibrillation, we were able to determine that the range between the 2 pRR intervals was the range of the optimal ventricular rate. A narrower range of the 2 pRR intervals was observed in patients with dilated cardiomyopathy than in the patients with no underlying cardiac disease. We conclude that it may be possible to determine the optimal ventricular rate in patients with atrial fibrillation by evaluating ventriculoarterial coupling. PMID- 12075865 TI - Autotransplantation of the heart for primary cardiac malignancy: development and surgical technique. AB - Primary cardiac malignancy presents an unusual and difficult surgical challenge. Malignant tumors of the left atrium have proved problematic due to their posterior location and difficulty of surgical exposure. The technique of cardiac explantation, ex vivo resection and cardiac reconstruction, and reimplantation- the cardiac autotransplantation procedure--was developed to solve this anatomic problem. Herein, we discuss the development of this approach and describe the surgical technique. PMID- 12075866 TI - Burckhard F. Kommerell and Kommerell's diverticulum. AB - Burckhard Friedrich Kommerell's scholarly description of the aortic diverticulum that bears his name was published in 1936. In the international literature, however the name of Kommerell survives only as an eponym. We present biographical information about Kommerell, as supplied by family members, and comment on the surgical relevance of his 1936 report. PMID- 12075867 TI - Active lymphocytic myocarditis treated with murine OKT3 monoclonal antibody in a patient presenting with intractable ventricular tachycardia. AB - This report describes the case of a 33-year-old woman with biopsy-proven, active lymphocytic myocarditis manifested by intractable ventricular tachycardia, nonspecific intraventricular block, and myocardial dysfunction. We treated hersuccessfully with OKT3 monoclonal antibody and antiarrhythmic agents. Immunosuppression is not recommended in patients with infectious or postinfectious myocarditis. However, it may have an important role in autoimmune myocarditis. In the few reports in the medical literature that we were able to find, OKT3 monoclonal antibody was administered early in the setting of acute, fulminant autoimmune myocarditis. Our patient received OKT3 therapy in a later phase of the disease, when inflammatory infiltrates were accompanied by extensive fibrosis and severe damage of cardiomyocytes. Our patient had concomitant Helicobacter pylori infection and a strong positive family history of gastric cancer, a disease often associated with H. pylori. We discuss the possibility of a causal relationship between H. pylori infection and autoimmune myocarditis. PMID- 12075868 TI - Interrupted aortic arch in an adult single-stage extra-anatomic repair. AB - Interrupted aortic arch is a rare congenital malformation of the aortic arch that occurs in 3 per million live births. Defined as a loss of luminal continuity between the ascending and descending portions of the aorta, this anomaly entails a very poor prognosis without surgical treatment. To our knowledge, the world medical literature contains only 12 reports of isolated interrupted aortic arch diagnosed in adults. Nine of these patients underwent successful surgical repair, but 1 died during the early postoperative period. We describe a 10th successful surgical repair, which involved a 42-year-old woman who had an asymptomatic type B interrupted aortic arch (characterized by interruption between the left subclavian and left carotid arteries). We performed a single-stage extra-anatomic repair by placing a 16-mm extra-anatomic Dacron graft between the ascending and descending portions of the thoracic aorta and by interposing a 7-mm extra anatomic Dacron graft between the 16-mm graft and the left subclavian artery. The patient recovered uneventfully and continued to do well 6 months later. PMID- 12075869 TI - Left ventricular pseudoaneurysm caused by coronary spasm, myocardial infarction, and myocardial rupture. AB - We report a very rare case of a 47-year-old man who had coronary spasm that resulted in a silent myocardial infarction, a ruptured myocardial wall, and a nonruptured left ventricular pseudoaneurysm. The patient presented with a 6-month history of dyspnea on exertion, without evidence of fixed coronary artery stenosis. Coronary angiography showed severe coronary spasm of the left anterior descending and left circumflex arteries; the spasm was relieved promptly by nitroglycerin. Echocardiography and left ventricular angiography revealed the large left ventricular pseudoaneurysm posterolateral to the left ventricle. We performed surgical resection of the pseudoaneurysm and patch repair of the ruptured left ventricular wall, with excellent results. We present this case because of the highly unusual sequence of events. Early surgical intervention resulted in the patient's recovery. PMID- 12075870 TI - Pseudoaneurysm of the popliteal artery: a rare sequela of acupuncture. AB - Acupuncture is an ancient Chinese treatment that has proved effective in relieving pain of various origins. Pseudoaneurysm resulting from acupuncture is an extremely rare event. Only 4 patients treated surgically for such a condition have previously been reported in the world medical literature. We present the case of a patient who had a pseudoaneurysm of the popliteal artery that resulted from acupuncture therapy for osteoarthritis of the knee. The patient was successfully treated by direct repair of the pseudoaneurysm. PMID- 12075871 TI - Protected iliofemoral venous thrombectomy in a pregnant woman with pulmonary embolism and ischemic venous thrombosis. AB - Although thromboembolism is uncommon during pregnancy and the postpartum period, physicians should be alert to the possibility because the complications, such as pulmonary embolism, are often life threatening. Pregnant women who present with thromboembolic occlusion are particularly difficult to treat because thrombolysis is hazardous to the fetus and surgical intervention by any of several approaches is controversial. A 22-year-old woman, in her 11th week of gestation, experienced an episode of pulmonary embolism and severe ischemic venous thrombosis of the left lower extremity The cause was determined to be a severe protein S deficiency in combination with compression of the left iliac vein by the enlarged uterus. The patient underwent emergency insertion of a retrievable vena cava filter and surgical iliofemoral venous thrombectomy with concomitant creation of a temporary femoral arteriovenous fistula. The inferior vena cava filter was inserted before the venous thrombectomy to prevent pulmonary embolism from clots dislodged during thrombectomy When the filter was removed, medium-sized clots were found trapped in its coils, indicating the effectiveness of this approach. The operation resolved the severe ischemic venous thrombosis of the left leg, and the patency of the iliac vein was maintained throughout the pregnancy without embolic recurrence. At full term, the woman spontaneously delivered an 8-lb, 6-oz, healthy male infant. PMID- 12075872 TI - Transaortic double valve replacement with total chordal preservation. AB - Very few cases of transaortic double valve replacement have been reported in the literature. A 26-year-old man presented to us with severe aortic regurgitation, mitral valve thickening, and mild mitral regurgitation 6 years after he had undergone a Ross procedure and open mitral commissurotomy. At his 2nd operation, he underwent transaortic double valve replacement with total chordal preservation of the mitral apparatus. Due to recurrent rheumatic activity, this patient had experienced a recurrence of valvulopathy Because we have observed this in other young patients with rheumatic heart disease, we no longer perform the Ross procedure in such patients, especially if there is associated mitral valve disease. In selected patients with dilated aortic annulus, the transaortic approach provides excellent access for safe mitral valve replacement with total chordal preservation. The surgical technique and a brief review of the literature are presented. PMID- 12075873 TI - Hemoptysis as an unusual presenting symptom of invasion of a descending thoracic aortic aneurysmal dissection by lung cancer. AB - A 70-year-old woman with a known chronic dissecting aneurysm of the descending thoracic aorta presented with new-onset back pain and hemoptysis. The hemoptysis was thought to be the result of invasion of the bronchial tree by the aneurysm. During surgical repair, a lesion that appeared to be a pulmonary abscess was discovered to be adhering to the aortic tissue, and the patient underwent a localized pulmonary resection. The pathology report of the surgical specimens revealed squamous cell carcinoma of the lung with infiltration of the aortic wall. The patient died of lung cancer 6 months later. Hemoptysis was an unusual presentation in a case of lung cancer that had invaded a stable chronic aortic aneurysm. PMID- 12075875 TI - Accidental perforation of the left ventricle with a chest draintube. PMID- 12075874 TI - Rapid ventricular tachycardias associated with cilostazol use. AB - A 92-year-old woman with normal systolic function had recently begun using the newly approved phosphodiesterase III inhibitor cilostazol when she was admitted with lower-extremity pain. Cilostazol is indicated for patients with intermittent claudication and contraindicated for patients with congestive heart failure. Two days after admission, the patient developed ventricular tachycardia. Cilostazol was discontinued, and shortly thereafter the ventricular tachycardia subsided. In this case, cilostazol was apparently an important predisposing factor for ventricular tachycardia. PMID- 12075876 TI - Massive pulmonary embolus with pre-existing inferior vena caval filter. PMID- 12075877 TI - Anomalous origin of the left coronary artery from the main pulmonary artery in two asymptomatic infants. PMID- 12075878 TI - Left coronary artery originating from the pulmonary artery in a 14-year-old girl. PMID- 12075879 TI - Are the channels too small in transmyocardial laser revascularization? PMID- 12075880 TI - Evidence of type A personality in a Chinese lady who died of acute myocardial infarction 2,100 years ago. PMID- 12075881 TI - Post-pneumonectomy chest tubes. PMID- 12075882 TI - Matters of the heart transcriptome: a brief history of cardiovascular genomics. PMID- 12075883 TI - Implantation of cryopreserved allograft pulmonary monocusp patch to treat nonthrombotic femoral vein incompetence. AB - We implanted cryopreserved allograft monocusp patches made from cadaveric pulmonary arteries to correct nonthrombotic valvular insufficiency of the common femoral vein in patients with chronic venous ulcers. Thirty-eight patients with 40 ulcers of longer than 3 years' duration underwent valvular repair of the common femoral vein from September 1995 through November 2001. We studied ulcer healing and competence of the monocusp patches using duplex ultrasonography Three patients with a total of 4 ulcers were lost to follow-up; therefore, 35 patients with 36 ulcers were available for examination. Twenty-four ulcers healed, and 12 did not. Of the 24 healed ulcers, 5 recurred. Four of the 5 ulcers were treated, 3 by monocusp patch replacement and 1 by iliac vein dilation and stenting. All such recurrent ulcers healed. The 5th recurrent ulcer was not treated, because the patient refused therapy Therefore, of the 24 ulcers that healed initially, 23 were healed at the end of the study Of the 12 unhealed ulcers, 4 were treated: 2 by ligation of incompetent perforating veins, 1 by saphenous vein ligation and partial stripping, and 1 by monocusp replacement. All 4 ulcers healed. The other 8 patients, all of whom had severe monocusp insufficiency, refused therapy. Therefore, of the 12 ulcers that did not heal initially, 4 were healed after additional treatment. No implanted monocusp patch developed clots. The ulcers remained healed when the prostheses remained competent. We conclude that monocusp patches can restore valvular competence to the femoral vein in patients who have primary valve insufficiency. In such patients, venous ulcers heal rapidly after prosthesis implantation if the monocusp remains competent and if there is no severe incompetence of the superficial venous system or of perforating veins. PMID- 12075884 TI - Dynamics of human postural transitions. AB - In the present study, the authors examined transitions between postural coordination modes involved in human stance. The analysis was motivated by dynamical theories of pattern formation, in which coordination modes and transitions between modes are emergent, self-organized properties of the dynamics of animal-environment systems. In 2 experiments, standing participants tracked a moving target with the head. Results are consistent with the hypothesis that changes in body coordination follow typical nonequilibrium phase transitions, exhibiting multistability, bifurcation, critical fluctuations, hysteresis, and critical slowing down. The findings suggest that posture may be organized in terms of dynamical principles and favor the existence of general and common principles governing pattern formation and flexibility in complex systems. PMID- 12075885 TI - Selective reaching: evidence for multiple frames of reference. AB - Students participated in 3 experiments investigating the use of environment- and action-centered reference frames in selective reaching. They pointed to a green target appearing either with or without a red distractor. Target-distractor distance was manipulated, and distractor interference (difference between distractor trials and no-distractor trials) was measured in reaction time, movement time, and movement endpoint. Target-distractor distance determined the dominant frame of reference. Small distances evoked an environment-centered framework that encoded targets within an external context. Large distances evoked an action-centered framework that encoded targets relative to the start position of the hand. Results support the hypothesis that the brain represents spatial information in multiple frames of reference, with the dominant frame of reference being dependent on the task demands. PMID- 12075887 TI - Naive optics: understanding the geometry of mirror reflections. AB - Paper-and-pencil tasks showed that many university students believed that when laterally approaching a mirror, they would see a reflection in the mirror before it was geometrically possible. Participants failed to adequately factor in the observer's location in the room. However, when asked about the behavior of a ray of light, participants knew about the law of reflection. No differences between psychology and physics students were detected, suggesting that the phenomenon is widespread and refractory to training. The findings were replicated with observers making judgments about image locations in a real room using a pretend mirror. Possible heuristics about mirror reflection that might explain the data are discussed. Naive optics is a promising venue to further knowledge of how observers understand basic laws of physics. PMID- 12075886 TI - Simultaneous dual-task performance reveals parallel response selection after practice. AB - E. H. Schumacher, T. L. Seymour, J. M. Glass, D. E. Kieras, and D. E. Meyer (2001) reported that dual-task costs are minimal when participants are practiced and give the 2 tasks equal emphasis. The present research examined whether such findings are compatible with the operation of an efficient response selection bottleneck. Participants trained until they were able to perform both tasks simultaneously without interference. Novel stimulus pairs produced no reaction time costs, arguing against the development of compound stimulus-response associations (Experiment 1). Manipulating the relative onsets (Experiments 2 and 4) and durations (Experiments 3 and 4) of response selection processes did not lead to dual-task costs. The results indicate that the 2 tasks did not share a bottleneck after practice. PMID- 12075888 TI - Understanding bias in proportion production. AB - The Stevens exponent (beta) can be obtained from proportion estimation judgments using the power model. In this article, the authors extend that model to proportion production, in which the relative magnitudes of 2 stimuli are adjusted to correspond to a numeric proportion (e.g., 1/4 or .25). The model predicts that when beta < 1, small proportions are underproduced, and large proportions are overproduced, but it predicts the reverse when beta > 1, which is the opposite of the predicted patterns for estimation. Eight participants estimated and produced magnitudes and proportions with spatial volume (beta < 1; Experiment 1) and color saturation (beta > 1; Experiment 2). The model's predictions were generally supported. An extension of the model using reference points can account for multicycle patterns shown by some participants. PMID- 12075889 TI - Dissociation of explicit and implicit timing in repetitive tapping and drawing movements. AB - Four experiments explored the hypothesis that temporal processes may be represented and controlled explicitly or implicitly. Tasks hypothesized to require explicit timing were duration discrimination, tapping, and intermittent circle drawing. In contrast, it was hypothesized that timing control during continuous circle drawing does not rely on an explicit temporal representation; rather, temporal control is an emergent property of other control processes (i.e., timing is controlled implicitly). Temporal consistency on the tapping and intermittent drawing tasks was related, and performance on both of these tasks was correlated with temporal acuity on an auditory duration discrimination task. However, timing variability of these 3 tasks was not correlated with timing variability of continuous circle drawing. These results support the hypothesized distinction between explicit and implicit temporal representations. PMID- 12075890 TI - Surface construal and the mental representation of scenes. AB - What distinguishes scenes from nonscenes? Photographs of objects on both naturalistic and blank backgrounds yielded boundary extension (BE: memory for unseen spatial expanse outside the picture's boundaries). However, line-drawn objects on blank backgrounds did not (Experiment 1). Perhaps the blank background was construed as depicting a real-world surface in the photograph condition but was construed as depicting nothing in the line-drawn condition. To change background construal, the authors used objects cut out of photographs; these were placed on blank backgrounds while viewers watched (Experiments 2 and 3). BE was eliminated. The authors propose that amodal continuation is a fundamental aspect of scene perception. However, not all pictures are scenes--only pictures construed as depicting a truncated view of a continuous world. PMID- 12075891 TI - Attentional processes in time perception: effects of mental workload and event structure. AB - The role of attention in timing was evaluated in 2 experiments. In Experiment 1, participants reproduced the durations of melodies with either a coherent or an incoherent structure. Participants were tested under control (timing only) and detection (timing plus target detection) workload conditions. Reproductions were shorter and more inaccurate under detection conditions, and incoherent event structure extended the effect to a wider range of durations. In Experiment 2, participants reproduced the durations of auditory prose passages that represented 3 levels of mental workload and 3 levels of event structure. Both increases in workload and the degradation of structure led to inaccurate reproductions. The results point to the central role of attention in temporal experience. PMID- 12075892 TI - Visual word recognition in bilinguals: phonological priming from the second to the first language. AB - In this study, the authors show that cross-lingual phonological priming is possible not only from the 1st language (L1) to the 2nd language (L2), but also from L2 to L1. In addition, both priming effects were found to have the same magnitude and to not be related to differences in word naming latencies between L1 and L2. The findings are further evidence against language-selective access models of bilingual word processing and are more in line with strong phonological models of visual word recognition than with the traditional dual-route models. PMID- 12075893 TI - Nonspatial attentional shifts between audition and vision. AB - This study investigated nonspatial shifts of attention between visual and auditory modalities. The authors provide evidence that the modality of a stimulus (S1) affected the processing of a subsequent stimulus (S2) depending on whether they shared the same modality. For both vision and audition, the onset of S1 summoned attention exogenously to its modality, causing a delay in processing S2 in a different modality. That undermines the notion that auditory stimuli have a stronger and more automatic alerting effect than visual stimuli (M. I. Posner, M. J. Nissen, & R. M. Klein, 1976). The results are consistent with other recent studies showing cross-modal attentional limitation. The authors suggest that such cross-modal limitation can be produced by simply presenting S1 and S2 in different modalities and that central processing mechanisms are also, at least partially, modality dependent. PMID- 12075894 TI - Fractionating the preview benefit in search: dual-task decomposition of visual marking by timing and modality. AB - Providing participants with a preview of half the distractors in a visual search task facilitates performance. The present study examined the effects of secondary tasks on the preview benefit in search. Participants had to attend to a visual or an auditory stream of digits that began either (a) at the onset of the preview or (b) after the preview. Secondary tasks that onset with the preview disrupted the preview benefit irrespective of their modality. Only visual secondary tasks disrupted the benefit in the delayed condition. These selective interference effects suggest that the preview benefit can be fractionated into 2 components: an initialization component that involves modality-independent resources and a maintenance component that depends on visual resources. Results are discussed in relation to theoretical accounts of the preview benefit in search. PMID- 12075895 TI - Orthographic neighborhood effects in lexical decision: the effects of nonword orthographic neighborhood size. AB - The effects of large neighborhoods (neighborhood size) and of higher frequency neighbors (neighborhood frequency) were examined as a function of nonword neighborhood size in lexical decision tasks. According to the multiple read-out model (J. Grainger & A. M. Jacobs, 1996), neighborhood size and neighborhood frequency effects should vary systematically as a function of nonword neighborhood size. In these experiments, the nonword context was more extensively manipulated than in previous studies, providing a more complete test of the model's predictions. In addition, simulations were conducted examining the model's ability to account for the facilitatory neighborhood size and neighborhood frequency effects observed in these experiments. The results suggest that the model overestimates the role of inhibition in the orthographic processing of English words. PMID- 12075896 TI - Masking by object substitution: dissociation of masking and cuing effects. AB - In a newly discovered form of visual masking, a target stimulus is masked by 4 flanking dots if their offset is delayed relative to the target (V. Di Lollo, J. T. Enns, & R. A. Rensink, 2000). In Di Lollo et al. (2000), the dot pattern also cued the relevant target and therefore required deliberate attention. In the present Experiments 2-6, a central arrow cued 1 of 2 letters for an E/F discrimination, with dots flanking both letters. Masking was reduced compared with the mask-cue procedure but was still robust. Delayed-offset dots flanking the nontarget also impaired performance, indicating competition for attention. Masking was unaffected by brightness of the dots relative to the target. Masking was attenuated not only by precuing attention to the target location but also by preview of an uninformative dot mask. Theories of masking by object substitution must therefore accommodate the prior context into which the target stimulus is introduced. PMID- 12075897 TI - Loci of signal probability effects and of the attentional blink bottleneck. AB - To investigate the locus of signal probability effects and the influence of stimulus quality on this locus, the authors manipulated probability in Task 2 of a psychological refractory period (PRP) paradigm. The effect was additive with stimulus onset asynchrony (SOA) when the target was not masked but underadditive with decreasing SOA when the target was masked. Even with masking, however, a range of probabilities had effects additive with SOA. The results suggest loci of stimulus probability before the PRP bottleneck as well as at or after the bottleneck. A second issue addressed was the locus of interference in the attentional blink (AB). The AB was larger when the probability of the first of 2 targets was lower. The results lead to the conclusion that one cause of the AB effect is a locus at least as late as the PRP bottleneck. PMID- 12075898 TI - Visual marking: selective attention to asynchronous temporal groups. AB - In visual search, when a subset of distractors is previewed 1 s before the target and the remaining distractors, search speed is independent of the number of previewed items. This is visual marking. What allows old items to be marked? Four experiments show that marking is disrupted if the onset of the new items is accompanied by synchronous changes to the old items, but it is not disrupted by changes restricted to the background or by asynchronous changes to the old items. Further, behaviorally relevant old items can be prioritized over new items. Visual marking is based on temporal asynchrony between new and old items, which allows segregation of these items into 2 temporal groups. Attention is then selectively applied to 1 group. PMID- 12075899 TI - Perceiving motion while moving: how pairwise nominal invariants make optical flow cohere. AB - Computer-generated sequences simulated observer movement toward 10 randomly placed poles, 1 moving and 9 stationary. When observers judged their direction of movement, or heading, they used 3 related invariants: The (a) convergence and (b) decelerating divergence of any 2 poles specified that heading was to the outside of the nearer pole, and the (c) crossover of 2 poles specified that heading was to the outside of the farther pole. With all poles stationary, the field of 45 pairwise movements yielded a coherent specification of heading. With I pole moving with respect to the others, however, the field could yield an incoherent heading solution. Such incoherence was readily detectable; similar pole motion leading to coherent flow, however, was less readily detectable. PMID- 12075900 TI - Forgive and remember. PMID- 12075901 TI - A comparison of bicortical and intramedullary screw fixations of Jones' fractures. AB - Two different fixations for treatment of Jones' fracture were tested in bone models and cadaveric specimens to determine the differences in the stability of the constructs. A bicortical 3.5-mm cannulated cortical screw and an intramedullary 4.0-mm partially threaded cancellous screw were tested using physiologic loads with an Instron 8500 servohydraulic tensiometer (Instron Corporation, Canton, MA). In bone models, the bicortical construct (n = 5, 87+/ 23 N) showed superior fixation strength (p = .0009) when compared to the intramedullary screw fixation (n = 5, 25+/-13 N). Cadaveric testing showed similar statistical significance (p = .0124) with the bicortical construct (n = 5, 152+/-71 N) having greater load resistance than the intramedullary screw fixation (n = 4, 29+/-20 N). In bone models, the bicortical constructs (23+/-9 N/mm) showed over twice the elastic modulus than the intramedullary screw fixations (9+/-4 N/mm) with statistical significance (p = .0115). The elastic modulus in the cadaveric group showed a similar pattern between the bicortical (19+/-17 N/mm) and intramedullary (9+/-6 N/mm) screw constructs. Analysis of the bicortical screw failure patterns revealed that screw orientation had a critical impact on fixation stability. The more distal the exit site of the bicortical screw was from the fracture site, the greater the load needed to displace the fixation. PMID- 12075902 TI - Biomechanical characteristics of the lateral ligament of the ankle joint. AB - The purpose of this study was to investigate the biomechanical properties of the lateral ankle ligaments and to correlate these properties with the distribution of mechanoreceptors in the ligaments reported previously. Biomechanical tensile tests were performed by using a video dimension analyzer to monitor the distortion of the lateral ligament at the pole and center in the animals. The mean upward slope of the load-elongation curves of the pole group was 38.1+/ 14.2, and that of the center group was 79.1+/-6.5. The pole of the ligaments tended to be elongated more than the center. These biomechanical results suggest that mechanoreceptors in the pole of the ligaments, compared with those in the center, may receive more mechanical stress and be effective in monitoring of tension applied to the ligament. PMID- 12075903 TI - A prospective comparison of clinical, radiographic, and intraoperative features of hallux rigidus: short-term follow-up and analysis. AB - Forty-seven patients (50 feet) underwent surgical intervention for symptomatic hallux rigidus between February 1998 and April 1999. Thirty-eight patients (41 feet) returned at 1 year for follow-up evaluation. Each foot was graded according to a four-stage hybrid radiographic grading system. At 1-year follow-up, 10 patients were classified as grade I, 17 as grade II, 12 as grade III, and 2 as grade IV. Subjective evaluation was based on a modified American Orthopaedic Foot and Ankle Surgery hallux metatarsophalangeal-interphalangeal 100-point scale. A pre- and postoperative objective physical examination and radiographic analysis were performed. Statistically significant differences between preoperative and postoperative values were found to exist for each portion of the subjective evaluation (p = .000); nonweightbearing dorsiflexion (p = .001); simulated weightbearing dorsiflexion (p = .003); metatarsal protrusion distance and angle of deviation of the second metatarsophalangeal joint (p = .000); and talar-first metatarsal angle (p = .015). For this specific patient population, the short-term results of surgical intervention for hallux rigidus provided subjective patient improvement and satisfaction, as well as a statistically significant but functionally minimal increase in first metatarsophalangeal joint dorsal range of motion. Additionally, in the 19 patients who underwent a periarticular decompression osteotomy, the intended correlation of plantar transposition of the capital fragment and offsetting the longitudinal shortening of the first metatarsal did not exist. PMID- 12075904 TI - Extracorporeal shock wave therapy for the treatment of chronic plantar fasciitis: indications, protocol, intermediate results, and a comparison of results to fasciotomy. AB - A review of the history, mechanism of action, and application of extracorporeal shock wave therapy for chronic plantar fasciitis is presented. The results of 40 feet treated with this modality are reviewed after a mean follow-up time of 8.4 months. All procedures were performed under intravenous sedation and local infiltrative anesthesia. An electrohydraulic shock wave with a mean of 20.6 kV combined with a mean of 2,506 pulses was used. The results of a similar demographic class of patients having undergone a percutaneous plantar fasciotomy at our institution were compared to the results of this cohort of shock wave patients. Eighty-two percent of the patients treated with extracorporeal shock wave therapy were successfully treated as compared to 83% with a percutaneous plantar fasciotomy. The mean score on the 11-point visual analog scale for satisfied patients was 7.9 preoperatively and 2.95 within 7 days postoperatively. After 3 months, the mean visual analog score was 4.2 or 50% of the preoperative value after a mean of 8.4 months following treatment. Eighty-three percent of the patients treated stated that shock wave therapy improved their symptoms. There were no complications encountered in any patient in this study. Extracorporeal shock wave therapy is an effective treatment, which significantly reduces the symptoms associated with chronic plantar fasciitis and compares favorably to the results achieved with surgical intervention in the form of a percutaneous plantar fasciotomy. PMID- 12075905 TI - Tendon interpositional arthroplasty of the fourth-fifth metatarsocuboid joint. AB - The most common complaints of patients with tarsal-metatarsal arthritis are severe loss of function and painful ambulation. In cases of failed conservative management, the recommended treatment is arthrodesis of the affected joint(s). This article presents a report of three cases of fourth-fifth metatarsocuboid arthritis treated with a fourth-fifth metatarsocuboid interposition alarthroplasty. Follow-up evaluation of all three patients demonstrated satisfactory pain relief. No postoperative complications were noted in any of the patients. Although more study is needed to evaluate the use of fourth-fifth metatarsocuboid tendon interpositional arthroplasty, the procedure offers promise and a suitable alternative to arthrodesis. PMID- 12075906 TI - Chondroblastoma of the talus: a case report. AB - Chondroblastoma of bone is an uncommon condition that has a predilection for secondary ossification centers, particularly of the humerus, tibia, and femur. Treatment usually involves surgical curettage and autogenous bone graft. This article reports the case of a chondroblastoma affecting the talus of a young man and a minimally invasive surgical approach for the treatment of this lesion. The patient presented with an 18-month history of right ankle pain and swelling. The lesion involved most of the talar body, with 2.5-cm cortical erosion inferiorly at the posterior talocalcaneal facet. A lateral approach was made to the talus and a window was fashioned at the distal end of the sinus tarsi under fluoroscopic guidance to allow access to the lesion. This was curetted and the defect was filled with autogenous bone graft taken from the ipsilateral iliac crest. The patient underwent an uneventful recovery, and was asymptomatic at follow-up 24 months postsurgery. He had 75% movement at the subtalar joint, when compared with the contralateral foot. PMID- 12075907 TI - Atypical coronal or sagittal Z ruptures of the achilles tendon: a report of four cases. AB - A typical rupture of Achilles tendon occurs as a complete rupture about 4-6 cm proximal to its insertion to the calcaneus. The morphology of the tear is most often random without any distinct recognizable pattern. The authors describe four cases of atypical "Z" rupture of the Achilles tendon. In two, the orientation of the "Z" was in the coronal plane and other two in the sagittal plane. All four cases were treated by open repair. PMID- 12075908 TI - Pigmented dermatofibrosarcoma protuberans of the foot with fibrosarcomatous changes: a review and case presentation. AB - Pigmented dermatofibrosarcoma protuberans, also known as Bednar's tumor, is a rare, aggressive, cutaneous tumor. The lesion is highly invasive but metastasis is rare. This article presents a case of Bednar's tumor of the foot with fibrosarcomatous changes and a review of the differential diagnoses of this lesion. PMID- 12075909 TI - Surgical approaches for hallucal sesamoid excision. PMID- 12075910 TI - Percutaneous K-wire fixation: protection of the exposed end. PMID- 12075911 TI - Strength comparison of allogenic bone screws, bioabsorbable screws, and stainless steel screw fixation. PMID- 12075912 TI - Do family physicians and internists differ in knowledge, attitudes, and self reported approaches for depression? AB - OBJECTIVES: The purpose of this investigation was to assess the relationship of primary care specialty training with self-assessed skill, knowledge, attitudes, and behavior toward depression recognition and management. METHOD: A baseline self-report questionnaire was administered to 184 internists and 138 family physicians participating in a multisite depression intervention study. RESULTS: There were no marked differences in knowledge of internists and family physicians regarding depression, in attitudes about the effectiveness of specific therapies, or in barriers to providing optimum treatment for depression. However, compared to internists, family physicians rated themselves as more skilled in the management of depression. When considering management of patients with moderate to severe depression, family physicians were more likely to report that they prescribed a selective serotonin-reuptake inhibitor (relative odds (RO) = 3.51, 95 percent Confidence interval (CI) [2.19, 5.60] and to personally counsel patients (RO = 1.97, 95 percent CI [1.16, 3.38]) more than half the patients, but were less likely to refer to a specialist in mental health (RO = 0.52, 95 percent CI [0.33, 0.82]) than were internists. Additional potentially influential characteristics did not wholly account for the reported differences in practice according to specialty. Physicians of both specialties expressed considerable uncertainty in their knowledge of psychotherapy and in their evaluation of the effectiveness of other strategies for the prevention of recurrence of depression. CONCLUSION: Strategies to improve mental health care should account for the orientation of primary care physicians to mental health issues. PMID- 12075913 TI - Lithium-induced tremor treated with vitamin B6: a preliminary case series. AB - OBJECTIVE: The occurrence of tremor in patients receiving lithium is well known, but the management of this side effect is a significant problem both for patients and physicians. Although some reports have suggested that beta-blockers may be useful in treating lithium-induced tremor (LT), these agents have different side effects which limit the possibility of their use. Vitamin B6 has been reported to be effective in treatment of patients suffering from different kinds of neuroleptic-induced movement disorders including parkinsonism and tardive dyskinesia. METHODS: This report presents the results of a preliminary four-week open-label clinical trial of five patients who suffered from LT and who were treated with vitamin B6 (900-1200 mg/d). The severity of tremor was assessed using the tremor subscale from the Simpson-Angus Scale (SAS) and Subjective Clinical Improvement Impression scale (SCII). RESULTS: After the addition of vitamin B6 to their treatment, according to the SAS scores four patients showed an impressive improvement until total disappearance of tremor. The subjective scale, on which the patients' scored their impression of clinical improvement, showed similar results. None of the patients suffered from any side effects attributable to vitamin B6. CONCLUSIONS: The results suggest that vitamin B6 may alleviate LT, double-blind controlled trials are needed to establish this effect. PMID- 12075914 TI - Physician willingness to refer older depressed patients for psychotherapy. AB - OBJECTIVE: Psychotherapy for late-life depression is an efficacious treatment option for older primary care patients who do not wish to take or do not respond to antidepressant medication. However, rates of physician referral to psychotherapy to treat late-life depression tend to be low. The purpose of this study was to assess attitudes toward psychotherapy for late-life depression and to identify predictors of physician willingness to refer older patients to psychotherapy. METHODS: Two hundred and five physicians identified from PPO directories of general internists in California and North Carolina completed a brief mailed survey about how they would treat a hypothetical older depressed patient and specific attitudes and practices regarding their own treatment of late-life depression. RESULTS: Only 27 percent of physicians said they would refer a depressed older patient to psychotherapy. In a regression analysis, female gender, the belief that psychotherapy is effective for older adults, and physician use of psychosocial techniques were associated with increased willingness to refer to psychotherapy. Practicing in North Carolina, awareness of depression treatment guidelines, and the perception of patient willingness to attend psychoeducational classes on depression and medication management were associated with decreased willingness to refer. CONCLUSIONS: More efforts are needed to increase the use of referral to psychotherapy as a treatment option for older medical patients. Education about guideline-level treatment alone may not be sufficient. More specific education, including information about the efficacy of psychotherapy for older adults, as well as direct training in psychosocial techniques, may be helpful in promoting referral to psychotherapy. PMID- 12075915 TI - Expressive disclosure and health outcomes in a prostate cancer population. AB - OBJECTIVE: This pilot study explored the feasibility and the efficacy of a brief, well-defined psychosocial intervention (expressive disclosure) in improving behavioral, medical, immunological, and emotional health outcomes in men with diagnosed prostate cancer. METHOD: Thirty prostate cancer patients receiving outpatient oncology care were randomized into experimental (disclosure) and control (non-disclosure) groups. All had been previously treated by surgery or radiation within the last 4 years and were being monitored without further intervention for change in PSA levels. Psychological and physical health surveys were administered and peripheral blood for PSA levels and immune assays was obtained upon study enrollment and again at 3 and 6 months post enrollment. Multivariate analyses were used to examine how the expressive disclosure impacted the hypothesized domains of functioning: physical and psychological symptoms; health care utilization; and immunocompetence. RESULTS: Compared to controls, patients in the expressive disclosure condition showed improvements in the domains of physical symptoms and health care utilization, but not in psychological variables nor in disease relevant aspects of immunocompetence. CONCLUSIONS: Study results support the feasibility of an expressive disclosure intervention for men with prostate cancer. The intervention was well accepted by this population, and participation/adherence was quite high. Results provide only limited support for the hypothesis that a written emotional disclosure task can positively impact health outcomes in a cancer population. However, this pilot study may have lacked adequate power to detect possible intervention benefits. Further studies with larger samples are needed to better assess the intervention's impact on psychological well-being and immunocompetence. PMID- 12075916 TI - Apolipoprotein E genotype and cognitive impairment in community-dwelling black older adults. AB - OBJECTIVE: The relationship between the epsilon 4 allele of the apolipoprotein E gene (APOE-epsilon4) located on chromosome 19 and Alzheimer's disease is well documented among Caucasian populations. However, the findings of research addressing the link between APOE polymorphism and neurocognitive functioning in populations of African origin from around the world have been equivocal. Therefore, the current study explored the relation of APOE-epsilon4 with cognitive impairment in a sample of community-dwelling English-speaking elderly blacks. METHODS: All participants (N = 57) were recruited consecutively from a community memory-screening program at a University affiliated Memory Disorders Clinic and evaluated using standardized assessment procedures. Cognitive impairment was classified according to an age and education adjusted Mini-Mental State Exam score of less than 24 as well as poorer functioning on a measure of delayed verbal memory. RESULTS: Increased risk for global cognitive dysfunction (OR = 9.5, 95 percent CI = 2.3-55.3, p = .004) and poorer verbal recall performance (beta = -.36, p = .006) were linked with the APOE epsilon4 allele after controlling for the potentially confounding effects of age, education, and gender. CONCLUSIONS: This investigation supports the role of APOE polymorphism in determining neurocognitive impairment among black elders residing in the community. PMID- 12075917 TI - Religious attendance and cause of death over 31 years. AB - OBJECTIVE: Frequent attendance at religious services has been reported by several studies to be independently associated with lower all-cause mortality. The present study aimed to clarify relationships between religious attendance and mortality by examining how associations of religious attendance with several specific causes of death may be explained by demographics, socioeconomic status, health status, health behaviors, and social connections. METHOD: Associations between frequent religious attendance and major types of cause-specific mortality between 1965 and 1996 were examined for 6545 residents of Alameda County, California. Sequential proportional hazards regressions were used to study survival time until mortality from circulatory, cancer, digestive, respiratory, or external causes. RESULTS: After adjusting for age and sex, infrequent (never or less than weekly) attenders had significantly higher rates of circulatory, cancer, digestive, and respiratory mortality (p < 0.05), but not mortality due to external causes. Differences in cancer mortality were explained by prior health status. Associations with other outcomes were weakened but not eliminated by including health behaviors and prior health status. In fully adjusted models, infrequent attenders had significantly or marginally significantly higher rates of death from circulatory (relative hazard [RH] = 1.21, 95 percent confidence interval [CI] = 1.02 to 1.45), digestive (RH = 1.99, p < 0.10, 95 percent CI = 0.98 to 4.03), and respiratory (RH = 1.66, p < 0.10, 95 percent CI = 0.92 to 3.02) mortality. CONCLUSIONS: These results are consistent with the view that religious involvement, like high socioeconomic status, is a general protective factor that promotes health through a variety of causal pathways. Further study is needed to determine whether the independent effects of religion are mediated by psychological states or other unknown factors. PMID- 12075918 TI - Late onset anorexia nervosa: two case reports. AB - OBJECTIVE: Some patients develop an eating disorder at a late age. It is worth investigating why this syndrome develops at such widely different ages. METHODS: Two Japanese cases of late onset anorexia nervosa are reported. RESULTS: In these late onset cases, anorexia developed out of sadness over a loss such as bereavement, divorce, and fear or anxiety for the future. In addition, these two patients didn't demonstrate a fear of aging or of losing their attractiveness and sexuality, nor did they develop a dissociative disturbance based on an early traumatic history of sexual abuse in PTSD. CONCLUSION: Although the clinical features are similar, there appear to be differences between late onset and adolescent cases in psychological mechanisms. PMID- 12075919 TI - Sickle cell disease and posttraumatic stress disorder. AB - Sickle Cell Disease (SCD) is a common condition among African Americans. It is associated with severe complications including severe pain in the chest, back, abdomen, or extremities. Individuals with SCD also have a reduced life span. Post traumatic stress disorder (PTSD) is a condition increasingly being recognized. In this article we discuss, to our knowledge, the first case of a patient with comorbid sickle cell disease and post-traumatic stress disorder. PMID- 12075920 TI - Separation and detection of neuroactive steroids from biological matrices. AB - This review is based on a selection of research papers published mainly in the last decade and it describes various analytical aspects of separation and detection of neuroactive steroids in biological matrices. PMID- 12075921 TI - Determination of sulfonylurea herbicides by continuous-flow liquid membrane extraction on-line coupled with high-performance liquid chromatography. AB - On-line coupling continuous-flow liquid membrane extraction (CFLME) with HPLC, a novel automatic system was developed for the determination of sulfonylurea herbicides in water. After an automatic trace-enrichment process by CFLME, which is the combination of continuous flow liquid-liquid extraction and support liquid membrane (SLM) extraction, the target analytes were concentrated in 50 microl of 0.2 M Na2CO3-NaHCO3 (pH 10.0) buffer. The concentrated sample solutions were injected directly onto a C18 analytical column with a valve, and detected at 240 nm with a diode array detector. Metsulfuron methyl (MSM), and DPX-A 7881 were baseline separated with a mobile phase consisting of methanol and 67 mM KH2PO4 Na2HPO4 (pH 5.91) buffer (45+55, v+v) at a flow-rate of 1.0 ml min(-1). With an enrichment time of 10 min and enrichment sample volume of 20 ml, the enrichment factors and detection limits are 100 and 0.05 microg l(-1) for MSM, and 96 and 0.1 microg l(-1) for DPX-A 7881, respectively. The linear range and precision (RSD) are 0.1-50 microg l(-1) and 7.0% for MSM, and 0.2-50 microg l(-1) and 9.2% for DPX-A 7881, respectively. This proposed method was applied to determine MSM and DPX-A 7881 in seawater, tap water, and bottled mineral water with spiked recoveries in the range of 83-95% for MSM and 88-100% for DPX-A 7881, respectively. PMID- 12075922 TI - Ion-exchange separation of proteins by polyallylamine-grafted cellulose gel. AB - A cellulose-based anion exchanger bearing water-soluble polycation was tested for separation of proteins. The exchanger was obtained by partial oxidation of cellulose gel by aq. NaIO4 followed by Schiff base formation with polyallylamine (PAA, molecular mass 5000). The retention behavior of proteins for three grades of PAA-cellulose gels, with amino group contents of 0.35, 0.59 and 0.96 mmol/g cellulose, was examined at several pH values and compared with that for conventional DEAE-cellulose gel with amino group content of 1.07 mmol/g cellulose. The retention of proteins by PAA-cellulose gels was remarkably greater than that for the DEAE-cellulose gel. Pairs of proteins having close isoelectric points and molecular masses (human and bovine serum albumins; beta-lactoglobulin A and B) could be separated by the PAA-cellulose gel columns. Such efficiency can be ascribed to high local density of grafted polyallylamine, in contrast to the random and sparse charge distribution in DEAE-cellulose. PMID- 12075923 TI - Role of the Na+ ion on phenol derivatives/hydroxypropyl-beta-cyclodextrin complex formation on porous graphitic carbon phase. AB - The reversed-phase liquid chromatography retention of phenol derivatives was investigated over a concentration range of sodium chloride (0-10(-2) M) and hydroxypropyl-beta-cyclodextrin (HP-beta-CD) (0-35x10(-3) M) using a porous graphitic carbon (PGC) stationary phase and a methanol/water mixture (50:50 (v/v)) as the mobile phase. A theoretical treatment was developed to investigate the effect of the sodium chloride and hydroxypropyl-beta-cyclodextrin on the equilibrium between the solutes with the PGC surface and the aqueous medium, respectively. The thermodynamic parameter variations were calculated using van't Hoff plots. It was expected that the sodium ion acted on the solute-PGC association process by modifying the surface tension of both the bulk solvent and the PGC surface. The phenol derivative/HP-beta-cyclodextrin complexation was shown to be entropically controlled for all the solutes except for the one which contained the -NO2 group in its structure, i.e. the nitro phenol derivative. A comparison of the compensation temperature of the solute-PGC association process when sodium chloride and HP-beta-CD concentration changed in the mobile phase led to the conclusion that these two modifiers acted via a variation in the hydrophobic effect. PMID- 12075924 TI - Rapid determination of minority organic acids in honey by high-performance liquid chromatography. AB - A rapid high-performance liquid chromatographic method for the determination of organic acids in honey is reported. Malic, maleic, citric, succinic and fumaric acids were identified and quantified in 15 min. First time repeatibility, reproducibility and recoveries were determined out for these acids in honey samples. Maleic acid was also quantified for first time by a chromatographic method. The organic acids were removed from honey by using a solid-phase extraction procedure with anion-exchange cartridges. Previously, the solution of honey was adjusted to pH 10.50 with 0.1 M NaOH and stirred for 15 min at room temperature. Then, this solution was adjusted to pH 5.00 with 0.1 M H2SO4. This procedure was carried out to avoid interferences in the baseline. The chromatographic separation was achieved with only one Spherisorb ODS-2 S5 column thermostated at 25 degrees C. Metaphosphoric acid (pH 2.20) was used as mobile phase at a flow-rate of 0.7 ml/min. Organic acids were detected with a UV-vis detector (215 nm). The precision results showed that the relative standard deviations of the repeatability and reproducibility were < or =3.20% and < or =4.86%, respectively. The recoveries of the organic acids ranged from 62.9 to 99.4%. Under optimum conditions the detection limits ranged from 0.0064 to 7.57 mg/kg and the quantification limits ranged from 0.025 to 10.93 mg/kg. PMID- 12075925 TI - Harmful azo colorants in leather. Determination based on their cleavage and extraction of corresponding carcinogenic aromatic amines using modern extraction techniques. AB - This study concerns the possibilities of using microwave-assisted extraction (MAE) or supercritical fluid extraction (SFE) for detection of harmful azo colorants in leather. After degreasing of the leather sample with SFE there follows a reductive cleavage of the azo colorants to their corresponding aromatic amines in the MAE or SFE equipment. The aromatic amines are subsequently extracted using either MAE or SFE and then finally determined by liquid chromatography with diode-array detection. The results have been compared with recoveries obtained using the German DIN method 53316. This standard method, based on conventional solvent extraction, is used in several European countries. Overall much better recoveries were obtained using MAE or SFE. With both MAE and SFE the amine recoveries of spiked leather samples were generally above 50%. The average recoveries were 62% for MAE and 60% for SFE (solvent collection) compared to 24% with the DIN method. For genuine leather samples the recoveries decreased, especially for benzidine. In this case the average values for MAE, SFE and DIN were 54, 38 and 19%, respectively. The quantification limits in leather samples using MAE or SFE were below 1 mg/kg for all amines investigated. The within laboratory precision was generally better than 10%, varying somewhat with the analyte considered. With the proposed methodology, the amount of hazardous organic solvents used could be decreased and the sample throughput increased with at least a factor of two with less manual handling compared to the DIN method. PMID- 12075926 TI - Reversed-phase high-performance liquid chromatography method for the determination of prolactin in bacterial extracts and in its purified form. AB - Reversed-phase high-performance liquid chromatography methodology for the determination of human prolactin (hPRL) in bacterial periplasmic space or in purified preparations has been developed. The technique, based on the high hydrophobicity of the hPRL molecule, allows its separation from the bulk of bacterial proteins. The precision for periplasmic shock fluid analysis was characterized by relative standard variations of 3-7% for intra-day and of 3-25% for inter-day determinations. Accuracy, evaluated by recovery tests, was of the order of 90%, a calibration curve being constructed with the use of a lyophilized osmotic shock fluid extract, which provided a stable, readily prepared internal reference. Sensitivity was of the order of 0.5 microg of hPRL. The methodology developed also provided a tool for comparing the hydrophobicity of glycosylated and non-glycosylated prolactin molecules obtained from several different species and of different preparations of native or biosynthetic human prolactin. PMID- 12075927 TI - Use of a linear gradient flow program for liquid chromatography-mass spectrometry protein-binding studies. AB - A rapid screening method to measure drug-protein binding using an immobilized human serum albumin (HSA) column was developed. This method utilizes a linear gradient flow-rate to accelerate the elution of strong binders to the HSA column. Post-column addition of a pressure relief valve enables mass spectrometric detection at relatively high mobile phase flow-rates (i.e., 2 ml/min). PMID- 12075928 TI - Liquid chromatographic determination of toxigenic secondary metabolites produced by Fusarium strains. AB - Various liquid chromatographic methods used in the analysis of mycotoxins (zearalenone, trichothecenes and fumonisins) produced by Fusarium species were compared in this work. The results demonstrate the suitability of modern clean-up procedures employing multifunctional MycoSep and immunoaffinity columns although these methods are more expensive than conventional methodologies for clean-up. HPLC with both fluorescence and photodiode array detection is a suitable technique for the analysis of toxic secondary metabolites produced by Fusarium species; different derivatisation strategies have been studied to improve the sensitivity of the technique because of the low concentration of these metabolites in contaminated food. The utility of the proposed methodology was assessed in cereal cultures of various Fusarium strains. PMID- 12075929 TI - Studies on separation of rare earth elements on various types of anion-exchangers in the C3H7OH-7 M HNO3 systems. AB - This paper presents the possibilities of separation of yttrium(III) neodymium(III) and samarium(III)-neodymium(III) pairs by frontal analysis on strongly basic anion-exchangers Wofatit SBW, Wofatit SBK and Lewatit MP 5080 in the systems: 90% (v/v) 2-propanol-10% (v/v) 7 M HNO3 and 90% (v/v) 1-propanol-10% (v/v) 7 M HNO3. The best results are obtained on Lewatit MP 5080 in the 90% (v/v) 2-propanol-10% (v/v) 7 M HNO3 system. In this system, on 1 dm3 of this anion exchanger, 1.95 kg yttrium(III) and 0.3 kg samarium(III) were purified decreasing the microcomponent content to below 10(-3)%. PMID- 12075930 TI - Field and flow programming in frit-inlet asymmetrical flow field-flow fractionation. AB - The separation of wide molecular mass (Mr) ranges of macromolecules using frit inlet asymmetrical flow field-flow fractionation (FI-AFlFFF) has been improved by implementing a combination of field and flow programming. In this first implementation, field strength (governed by the cross flow-rate through the membrane-covered accumulation wall) is decreased with time to obtain faster elution and improved detection of the more strongly retained (high Mr) materials. The channel outlet flow-rate is optionally held constant, increased, or decreased with time. With circulation of the flow exiting the accumulation wall to the inlet frit, the dual programming of cross flow and channel outlet flow could be implemented using just two pumps. With this flow configuration, the channel outlet flow-rate is always equal to the channel inlet flow-rate, and these may be programmed independently of the cross flow-rate through the membrane. FI-AFlFFF retains its operational advantage over conventional asymmetrical flow FFF (AFlFFF). Unlike conventional AFlFFF, FI-AFlFFF does not require time consuming, and experimentally inconvenient, sample focusing and relaxation steps involving valve switching and interruption of sample migration. The advantages of employing dual programming with FI-AFlFFF are demonstrated for sets of polystyrene sulfonate standards in the molecular mass range of 4 to 1000 kDa. It is shown that programmed FI-AFlFFF successfully expands the dynamic separation range of molecular mass. PMID- 12075931 TI - Simultaneous modeling of the Kovats retention indices on OV-1 and SE-54 stationary phases using artificial neural networks. AB - In this study, a quantitative structure-property relationship technique has been used for the simultaneous prediction of Kovats retention indices for some esters, alcohols, aldehyde and ketones on OV-1 and SE-54 stationary phases, using an artificial neural network (ANN). The best-selected descriptors that appear in the models are the molecular values, number of atoms in each molecule, molecular shadow area on the xy plane and the energy level of the highest occupied molecular orbital. A 4-6-2 ANN was generated using these descriptors as inputs and its outputs will be the Kovats retention indices on OV-1 and SE-54 stationary phases. After optimization of the network parameters, the network was trained using a training set. For the evaluation of the predictive power of the generated ANN, an optimized network was used to predict the Kovats retention indices of the prediction set. The results obtained in this study showed that the average percentage deviation between the predicted ANN and the experimental values of Kovats retention indices for the prediction set were 2.5 and 3.0% on the OV-1 and SE-54 stationary phases, respectively. These values are in good agreement with the experimental results. PMID- 12075932 TI - Cloning of human myeloid-associated differentiation marker (MYADM) gene whose expression was up-regulated in NB4 cells induced by all-trans retinoic acid. AB - A full-length cDNA of 3192 bp isolated from human bone marrow cDNA library was predicted an ORF encoding 298 amino acids. The deduced protein, containing seven putative transmembrane segments and sharing 75.8% amino acid identity with mouse Myadm protein, was named as human MYADM. The results of Northern blot analysis showed that MYADM was ubiquitously expressed in 15 of 16 adult tissues tested, except thymus. To determine whether the novel human gene was involved in hematopoietic differentiation process as mouse Myadm did, we examined the mRNA expressive abundance of this gene between normal bone marrow cells and peripheral blood leukocytes, and detected the expression change in NB4 cells induced by all trans retinoic acid at different induce time by the semi-quantitative RT-PCR. The results showed that the expression of the novel gene was not only significantly higher in peripheral blood leukocytes than in bone marrow cells, but also significantly up-regulated when the NB4 cells(derived from a patient with acute promyelocytic leukemia) were induced by all-trans retinoic acid (ATRA) for 48hr. It is suggested that human MYADM was also associated with the differentiation of hematopoietic cells or acute promyelocytic leukemia cells. In addition, MYADM was mapped to human chromosome 19q 13.33-q 13.4 by Radiation Hybrid mapping, and it consists of 3 exons and 2 introns and spans a 7.1-Kb genomic region. PMID- 12075933 TI - Gene expression profiling in osteoblast biology: bioinformatic tools. AB - This review focuses on using microarray data on a clonal osteoblast cell model to demonstrate how various current and future bioinformatic tools can be used to understand, at a more global or comprehensible level, how cells grow and differentiate. In this example, BMP2 was used to stimulate growth and differentiation of osteoblast to a mineralized matrix. A discussion is included on various methods for clustering gene expression data, statistical evaluation of data, and various new tools that can be used to derive deeper insight into a particular biological problem. How these tools can be obtained is also discussed. New tools for the biologists to compare their datasets with others, as well as examples of future bioinformatic tools that can be used for developing gene networks and pathways for a given set of data are included and discussed. PMID- 12075934 TI - Structure of transfection-active histone H1/DNA complexes. AB - Relationships between the structure of transfecting complexes of histone H1 and DNA and their transfection efficiency were studied. Transfection activity proved to be connected to complex aggregates. Low speed centrifugation of the complexes resulted in loss of the transfection activity. The complexes/aggregates were active with high efficiency in a broad range of weight input ratios r(i) (0.1 < r(i) < 30). Using atomic force microscopy (AFM), the complexes were imaged at negative, nearly electroneutral and positive charge conditions. Electroneutral complexes at r(i) = 1 showed a multitude of different complex forms. Fibrillar, network-like and branched structures were frequently present in one complex. Strongly positive charged complexes had a toroidal appearance. All these different forms contributed to the high transfection efficiency. Cellular uptake is supposed to be by phagocytosis. PMID- 12075935 TI - Characterization of the antiapoptotic (p35) gene homologue of Spodoptera litura nucleopolyhedrosis virus (SlNPV). AB - Antiapoptotic genes of baculoviruses have been shown to prevent virus induced apoptosis in insect cells. Dot blot and Southern hybridizations of EcoRI genomic library and genomic digests of Spodoptera litura nucleopolyhedrosis virus (SlNPV) respectively give strong hybridization signals with antiapoptotic DNA (p35 gene) probe of the prototype Autographa californica nucleopolyhedrosis virus (AcNPV). Both the hybridizations indicate the presence of a homologous gene in the 1.8 kb EcoRI-Y fragment of SlNPV. The sequence of 1.244 kb region of this fragment encompasses an open reading frame coding for a polypeptide of 296 amino acids under sequential early (TATA) and late (TAAG) promoter motifs like that in other baculovirus p35 genes. The putative SlNPV p35 ORF expresses abundantly as a 35 kDa protein in Spodopterafrugiperda (Sf9) cells when allowed to express under the polyhedrin promoter of AcNPV. PMID- 12075937 TI - Masses of phosphorylated neurofilaments are associated with abnormal golgi apparatus of anterior horn neurons of beta, beta'-iminodipropionitrile intoxicated rats. AB - The Golgi apparatus (GA) of anterior horn neurons of rats chronically intoxicated with beta,beta'-iminodipropionitrile (IDPN) in drinking water was examined with an organelle-specific antibody. The neuropile of the anterior horns contained the typical axonal spheroids associated with IDPN toxicity while the perikarya of approximately one-third of the neurons contained phosphorylated neurofilaments, which are not found in the perikarya of control rat neurons. By serial or double immunostaining with the SMI-31 and anti-MG 160 antibodies, there were no morphological changes of the GA in the majority of neurons including neurons with a mild to moderate degree of neurofilamentous accumulation. However, a few neurons with a massive accumulation of phosphorylated neurofilaments contained abnormal profiles of the GA which consisted of focal clustering, reduction in size and fragmentation. The results suggest that masses of phosphorylated neurofilaments are associated with structural abnormalities of the GA. PMID- 12075938 TI - Immunohistochemical characterization of pineal parenchymal tumors using novel monoclonal antibodies to the pineal body. AB - To characterize the immunohistochemical nature of pineal parenchymal tumors (PPT), we examined nine cases of normal pineal bodies and 23 cases of PPT using several neuronal and glial antibodies and 10 novel monoclonal antibodies raised against human pineal tissue. The PPT were classified into four pineocytoma, five pineal parenchymal tumor of intermediate differentiation (Int-PPT), and 14 pineoblastoma. The pinealocytes, parenchymal cells of the pineal body, were labeled with five, neuronal and seven pineal monoclonal (from PP1 to PP7) antibodies in the normal pineal bodies. The subjects ranged from 3 to 85 years old, 12 female and eight male subjects were studied. Antibodies to glial cells PI1, PI2 and PX1, stained interstitial cells of the pineal body. Many of the PPT showed positive immunostaining for pinealocyte-associated antigens and neuronal markers. The intensity of immunostaining showed some association with the degree of differentiation of the tumor, but there was a considerable variety of staining from case to case. The pineocytomas are more immunopositive than are the Int-PPT or pineoblastoma for neuronal and pinealocyte-associated antibodies. In particular the neurofilament protein (NFP)68 kDa, PP1 and PP6 showed significant differences of reactivity between pineocytoma, Int-PPT and pineoblastoma, when compared in groups showing extensive positive staining (positive staining in almost all areas of the tumor). By using three representative antibodies, anti NFP68kDa, PP1 and PP6, we were able to make a clear distinction between pineoblastoma, Int-PPT and pineocytoma. Glial fibrillary acidic protein (GFAP), PI1 and PI2 antibodies only occasionally showed a small number of positive cells in the tumor, and thus we considered these cells to be non-neoplastic interstitial cells or reactive astrocytes entrapped in the tumor. Our data suggest that the glial differentiation of PPT may occur, but that it seems to be a very rare event. PMID- 12075936 TI - The regulatory network controlling beta-globin gene switching. AB - The human globin gene cluster, which represents a prototypical eukaryotic multigene locus, has been investigated for more than two decades and is classic model for coordinate control of tissue-specific gene expression. It is well known that globin gene expression is restricted to specific tissues and that globin genes are sequentially switched on during development. What intricate regulatory mechanisms account for tissue-specific transcriptional control of globin gene expression? Previous studies have focused on the interactions of trans-acting factors and cis-acting elements including the locus control region (LCR), which is considered a potent enhancer in globin gene switching. More recent studies have not only focused on the local DNA regulatory elements but also on remodelling of chromatin and transcription at the globin gene cluster within the native genomic context. Moreover, several studies have presented extensive data that address whether the LCR is required to open the chromatin. Although there is increased insight into the regulation of the beta-globin gene switching, many aspects relating to the developmental activation of distinct globin genes remain elusive. PMID- 12075939 TI - Parkinson's disease mimicking senile dementia of the Alzheimer type: a clinicopathological study of four autopsy cases. AB - This report concerns four Japanese autopsy cases of Parkinson's disease (PD) mimicking senile dementia of the Alzheimer type. Three patients with a clinical diagnosis of senile dementia of the Alzheimer type developed memory disturbance as the initial sign, and a patient with a clinical diagnosis of atypical senile dementia presented with hallucination and delusion as the initial sign. Dementia was evident in all four patients, and slight parkinsonism appeared in the middle to late stages of the disease in two patients. Macroscopical examination of the brain disclosed slight depigmentation of the substantia nigra and prominent depigmentation of the locus ceruleus in all four cases. Histological examination of the four patients showed neuronal loss with astrocytosis and the appearance of Lewy bodies in the substantia nigra, locus ceruleus, and dorsal vagal nucleus. The nucleus basalis of Meynert was involved in three cases, in which this structure was examined. The total Lewy body scores of the four cases were 1 in three cases and 0 in the other, compatible with PD. Massive appearance of senile plaques, consistent with Braak stage C, was found in one case, and the slight appearance of senile plaques, consistent with Braak stage A, was evident in two cases. One case had no evidence of senile plaques. In all four cases, slight neurofibrillary changes were present in the limbic areas, compatible with Braak stages II to III. Based on these clinicopathological findings and a review of the literature, we concluded that PD simulating Alzheimer's disease without overt parkinsonism rarely exists. Furthermore, we postulate that the clinical features of PD are more widespread than previously believed. PMID- 12075940 TI - Imaging and pathology in pediatric neurological disorders. AB - Radiologic and pathologic findings correlate well in many pediatric neurologic disorders. However, there are some exceptions. Radiologic techniques are unable to visualize or identify pathologic changes in some diseases (e.g. status marmoratus and subdural fluid collection), whereas pathologic examination fails to clarify the histologic substrate of radiologic abnormalities in other conditions (e.g. Fukuyama-type congenital muscular dystrophy and neurofibromatosis type 1). Although studies to solve these issues have met with practical difficulties, important progress has recently been made on some disorders such as Fukuyama-type congenital muscular dystrophy and acute necrotizing encephalopathy. PMID- 12075941 TI - Review of evidence for a connection between Chlamydia pneumoniae and atherosclerotic disease. AB - BACKGROUND: Established risk factors account for no more than 50% of coronary artery disease cases; therefore, the search continues for other modifiable risk factors. In recent years, there has been renewed interest in the infectious theory of atherosclerosis. Chlamydia pneumoniae has been implicated as a potential cause of atherosclerotic disease. OBJECTIVE: This review discusses possible mechanisms of C pneumoniae involvement in atherosclerosis, summarizes the case-control studies and antibiotic trials completed, and identifies remaining questions about future therapy. METHODS: Published data were identified by a MEDLINE search of the English-language literature from 1966 through 2001 using the terms Chlamydia, atherosclerosis, and coronary artery disease. Relevant conference presentations and book chapters were also included. RESULTS: C pneumoniae antibodies are found in approximately 50% of middle-aged adults world wide. These antibodies have been detected in atherosclerotic tissue by various methods, including microimmunofluorescence, and several studies have linked high antibody titers with increased risk of cardiovascular events. A few possible mechanisms for this perceived increase in risk have been proposed, such as induction of atheroma through damage to the endothelium, expression of procoagulant factor leading to thrombus formation, and production of cytokines resulting in increased inflammatory response. Results of animal studies suggest that early antibiotic treatment may reduce cardiovascular risk, but the first human studies have not produced conclusive results. CONCLUSIONS: Although a connection has been suggested, the precise mechanism by which C pneumoniae affects atherosclerosis has not yet been identified. Large-scale trials are needed to determine whether eradication of C pneumoniae reduces the incidence of cardiovascular events in humans. PMID- 12075942 TI - A controlled, double-blind, multicenter study comparing clarithromycin extended release tablets and levofloxacin tablets in the treatment of community-acquired pneumonia. AB - BACKGROUND: Macrolides and fluoroquinolones are frequently used for the empiric treatment of community-acquired pneumonia (CAP). OBJECTIVE: The aim of the study was to compare the safety profile and efficacy of clarithromycin extended-release (ER) tablets with those of levofloxacin tablets for the treatment of CAP in ambulatory adult patients. METHODS: In a Phase III, double-blind, randomized, parallel-group, multicenter study, ambulatory adult patients (> or = 18 years) with signs and symptoms of CAP received a 7-day course of treatment with either clarithromycin ER (two 500-mg tablets once daily) or levofloxacin (two 250-mg tablets once daily). A diagnosis of CAP was confirmed by radiography of the chest and physical examination, and sputum samples were analyzed to identify etiologic pathogen(s). Tolerability was assessed through subjective reports of adverse events and through changes in physical findings, concomitant medications, and laboratory values. RESULTS: There were no statistically significant differences between treatment groups in terms of sex, age, race, or body weight. The mean age was 50 years (range, 18-91 years). Of 299 patients randomized and treated, 252 were clinically evaluable (128 clarithromycin ER, 124 levofloxacin). The 95% CI for the difference between cure rates demonstrated equivalence of the 2 treatments. Among clinically evaluable patients at the test-of-cure visit, clinical cure rates were 88% (113/128) and 86% (107/124), and radiographic success rates were 95% (117/123) and 88% (104/118) for clarithromycin ER and levofloxacin, respectively. Both treatment regimens were effective in resolving and improving clinical signs and symptoms of CAP. Among clinically and bacteriologically evaluable pa- tients, bacteriologic cure rates were 86% (80/93) and 88% (85/97) for clarithromycin ER and levofloxacin, respectively. No statistically significant differences were observed between the 2 treatment groups in the overall incidence of adverse events. CONCLUSIONS: Clarithromycin ER demonstrated equivalent efficacy and tolerability to the fluoroquinolone levofloxacin in a group of ambulatory adult patients with CAP. Clarithromycin ER also appeared to be safe in the population studied. PMID- 12075943 TI - Safety profile and tolerability of intravenous AR-C69931MX, a new antiplatelet drug, in unstable angina pectoris and non-Q-wave myocardial infarction. AB - BACKGROUND: Thrombin generation and platelet aggregation in the disrupted atherosclerotic plaque are the major reasons for thrombus formation associated with acute coronary events. AR-C69931MX is a new agent that inhibits adenosine diphosphate-induced platelet aggregation by antagonism of the P(2T) purinoceptor. OBJECTIVE: This study assessed the safety profile, tolerability, and plasma concentrations at steady state of intravenous AR-C69931MX in patients with unstable angina pectoris or non-Q-wave myocardial infarction (MI). METHODS: This was a Phase II, multicenter, double-blind, randomized, placebo-controlled trial. Patients with unstable angina or non-Q-wave MI were randomized to a 72-hour infusion of AR-C69931MX or placebo as adjunctive therapy to aspirin and low molecular-weight heparin. Other treatment was at the discretion of the local investigator. Outcomes were assessed at 30 days. RESULTS: Ninety-four patients were randomized and 91 received treatment (45 AR-C69931MX, 46 placebo). Plasma concentrations of AR-C69931MX were within the expected range, there were no signs of accumulation, and interindividual variability in clearance was low. Four patients receiving AR-C69931MX discontinued treatment due to minor bleeding events, and 5 patients receiving placebo discontinued treatment due to other adverse events or deterioration in their condition. No serious bleeding events were seen during treatment. The incidence of > or = 1 episode of minor bleeding was slightly higher in patients receiving AR-C69931MX compared with those receiving placebo (38% vs 26%, respectively). The drug was well tolerated hemodynamically, and there were no significant changes in other laboratory values between groups. CONCLUSIONS: As adjunctive therapy to aspirin and low-molecular weight heparin in patients with unstable angina or non-Q-wave MI, intravenous AR C69931MX was well tolerated, with no difference in the incidence of serious adverse events compared with placebo. PMID- 12075944 TI - Comparison of granisetron and ramosetron for the prevention of nausea and vomiting after thyroidectomy. AB - BACKGROUND: Postoperative nausea and vomiting (PONV) are common after thyroidectomy. Granisetron, a selective 5-hydroxytryptamine type 3 (5-HT3) receptor antagonist, is more effective than the traditional antiemetics droperidol and metoclopramide for the prevention of PONV after thyroidectomy. Ramosetron, another new selective antagonist of 5-HT3 receptor, has been shown to have more potent and longer-acting properties than granisetron against cisplatin induced emesis in ferrets. OBJECTIVE: This study was undertaken to compare the efficacy and tolerability of granisetron and ramosetron for the prevention of PONV after thyroidectomy. METHODS: In this randomized, double-blind study, patients received granisetron 3 mg or ramosetron 0.3 mg intravenously at the end of surgery. A standard general anesthetic technique and postoperative analgesia were used. Emetic episodes and safety were assessed during the first 24 hours and the second 24 hours after anesthesia. RESULTS: Eighty patients (15 men, 65 women) aged 33 to 58 years were included in the study. Each study group consisted of 40 patients. There were no differences between groups with regard to patient demographics. The percentage of patients who were emesis free (no nausea, no retching, no vomiting) 0 to 24 hours after anesthesia was 83% (33 of 40) with granisetron and 88% (35 of 40) with ramosetron; the corresponding rates 24 to 48 hours after anesthesia were 63% (25 of 40) and 90% (36 of 40), respectively (P = 0.004). The safety profile of the two 5-HT3 receptor antagonists was comparable, as no clinically serious adverse effects caused by the study drug were observed in either of the groups. CONCLUSIONS: Prophylactic antiemetic therapy with ramosetron was comparable to therapy with granisetron for the prevention of PONV 0 to 24 hours after anesthesia in patients who underwent thyroidectomy; 24 to 48 hours after anesthesia, ramosetron was more ef- fective than granisetron for prophylaxis against PONV in this population. PMID- 12075945 TI - A randomized, parallel, vehicle-controlled comparison of two erythromycin/benzoyl peroxide preparations for acne vulgaris. AB - BACKGROUND: Topical erythromycin/benzoyl peroxide (EBP), marketed for acne treatment, must be compounded by a pharmacist and requires subsequent refrigeration, warranting the development of alternate formulations. OBJECTIVE: This trial compared the efficacy and tolerability of a single-use EBP combination package (EBP Pak) with those of its matching vehicle control (VC Pak) and the original, reconstituted formulation packaged in a jar (EBP Jar). The matching VC for the original formulation (VC Jar) was used to achieve study blinding. METHODS: In this double-blind, parallel-group, multicenter study, patients were randomly assigned to the 4 treatment arms. The primary efficacy evaluations were lesion reductions from baseline and treatment success (as defined by a Physician's Global Acne Severity score of 0 [clear] or 0.5 [sparse comedones with few or no inflammatory lesions]). Secondary evaluations were Physician's Global Acne Severity scores, facial-oiliness scores, and end-point patient evaluations of global improvement and treatment acceptability. Tolerability was based on the incidence and severity of adverse events. RESULTS: Three hundred twenty-seven patients (age range, 12-46 years) were randomly assigned to the 4 treatment groups (EBP Pak, 124; VC Pak, 42; EBP Jar. 121; VC Jar, 40). Mean percent reductions in total acne lesions, inflammatory acne lesions, and come-dones from baseline were significantly greater with EBP Pak than with VC Pak (P < or = 0.001 for the intent-to-treat patient population after 8 weeks). Statistical significance for all lesion parameters was demonstrated at week 2 (P < 0.05) and maintained throughout the study. At 8 weeks, a significantly greater proportion of patients demonstrated treatment success with the EBP Pak compared with VC Pak (28% vs 2%, respectively; P < 0.001). The EBP Pak was comparable to the EBP Jar in terms of reduction in acne lesions, Physician's Global Acne Severity scores, and end-of-treatment patient evaluations of global improvement. No serious drug related adverse events were reported. CONCLUSIONS: Results of this 8-week trial demonstrate that the single-use combination package of EBP is well tolerated, effective, and comparable to the original formulation for the treatment of acne vulgaris in this selected patient population. PMID- 12075946 TI - A postmarketing, open-label study to evaluate the tolerability and effectiveness of replacing standard-formulation doxazosin with doxazosin in the gastrointestinal therapeutic system formulation in adult patients with hypertension. AB - BACKGROUND: The antihypertensive doxazosin works to decrease perivascular muscular tone, causing vasodilatation and hence a decrease in peripheral vascular resistance. To prevent the sharp decrease in blood pressure (BP), syncope, and other postural effects that may occur at the beginning of therapy with this drug, the dose must be adjusted. A new formulation, doxazosin gastrointestinal therapeutic system (GITS), allows slow release of the active agent so therapeutic serum levels are reached within 24 hours, rendering dose adjustment unnecessary and eliminating any first-dose effects. OBJECTIVES: The goals of this study were to evaluate the tolerability and effectiveness of (1) using doxazosin in the standard and new GITS formulations in adult patients with hypertension who either had uncontrolled or newly diagnosed disease, and (2) replacing standard formulation doxazosin with doxazosin in the GITS formulation. METHODS: This was a postmarketing, open-label, noncomparative, multicenter clinical study covering primary care patients diagnosed with essential uncontrolled arterial hypertension (AHT). Subjects could be patients who were undergoing drug therapy before enrollment or those diagnosed with AHT and/or treated for the disease for the first time on entering the study. The study covered a period of 6 to 9 months, divided into 2 phases. Phase 1 involved a minimum of 3 and maximum of 6 months of treatment with standard-formulation doxazosin. Phase 2 commenced with the changeover from standard-formulation doxazosin to the GITS formulation and lasted 12 weeks. The principal study variables included BP and the development of adverse events (AEs). At every visit, the patients were asked by an investigator whether they had suffered any AEs since the previous contact. RESULTS: Of the total of 4,512 patients initially enrolled, 3537 (78.4%) completed the study. A total of 285 patients were excluded for failing to comply with the inclusion criteria, leaving 4,227 patients for analysis. In most instances, premature withdrawal from the study (16.3% [690/4,227]) was due to loss to follow-up (37.2% [257/690]), followed by the development of AEs (27.8% [192/690]). Fifty-nine percent (2,493/4,226) of patients analyzed were men and 41.0% (1,733/4,226) were women (sex data not recorded for 1 patient), with a mean age of 62.4 years (SD, 10.6). Among the patients participating, 54.8% (2,316/4,227) presented with some type of associated disease. The percentage of patients undergoing monotherapy was 70.7% (2,987/4,227); the remainder (29.3% [1,240/4,227]) underwent a combined therapy regimen using another antihypertensive drug in conjunction with doxazosin. The mean initial systolic and diastolic BPs were 160 +/- 10.63 mm Hg and 95.26 +/- 7.21 mm Hg, respectively. Reduction in systolic BP was 20.9 mm Hg in phase 1 and 3.8 mm Hg in phase 2. In the case of diastolic BP, the reduction was 13.3 mm Hg in phase 1 and 2.6 mm Hg in phase 2. The percentage of patients with controlled disease was 47.9% (1,891/3,949) by the end of phase 1 and 63.4% (2,242/3,537) by the end of phase 2. A total of 322 (7.6%) patients presented with 343 AEs. 37 (0.9%) of which were deemed severe (0.6% in phase 1 and 0.3% in phase 2). Limitations included the following: (1) the design of this study did not allow comparison of the 2 formulations regarding effectiveness or tolerability: (2) the patients who remained in the study after phase 1 were those less susceptible to toxicity from doxazosin; and (3) it is probable that the nonresponders were more likely to drop out of the study. CONCLUSIONS: Doxazosin in the standard formulation was effective and well tolerated for the purpose of lowering BP. Patients who tolerated the standard formulation also tolerated the switch to the GITS formulation. Finally, this substitution did not negatively impact the effectiveness of treatment of AHT. PMID- 12075947 TI - The prescription drug user fee act: active postmarketing drug surveillance and safety. PMID- 12075948 TI - Clinical and economic outcomes associated with olanzapine for the treatment of psychotic symptoms in a county mental health population. AB - BACKGROUND: The comparatively high acquisition costs of the newer antipsychotic medications have caused the mental health community to look closely at their potential benefits. OBJECTIVE: The purpose of this study was to perform a naturalistic analysis of changes in mental health service utilization, economic costs, and clinical outcomes after the initiation of olanzapine therapy for psychotic symptoms in an indigent patient population from a large county-operated mental health care system. METHODS: This was a prospective, uncontrolled investigation using a mirror-image cohort design. All captured costs from patients who began olanzapine therapy between November 1, 1996, and April 30, 1998, were analyzed in an intent-to-treat fashion to compare resource utilization in the 12 months immediately before and after the intervention. Clinical function was assessed at baseline and 6 months using the Positive and Negative Syndrome Scale (PANSS). In a subgroup analysis, the baseline characteristics of patients who completed 12 months of olanzapine treatment were compared with those of patients who (1) changed medication or (2) changed pay or source or were lost to follow-up. RESULTS: One hundred eighty-nine patients were started on olanzapine treatment during the 18-month study entry phase. Patients were primarily male (63.5%) and had a mean age of 35.9 years. Most (66.3%) had a formal diagnosis of thought disorder. Fifty-six patients received olanzapine for 12 consecutive months, and 22 were switched to other psychotropic medications. Of the remaining 111 patients, 70 changed payors (ie, qualified for Medicaid), and 41 were lost to follow-up. In the subgroup analysis, patients who completed 12 months of treatment (ie, responders) had significantly lower mean PANSS total scores at baseline compared with those who changed payors or were lost to follow-up (P = 0.047), and were significantly more likely to have a formal diagnosis of thought disorder (P = 0.039). Responders demonstrated a significant reduction in PANSS total and negative subscale scores at 6-month follow-up (both measures, P < 0.001). In the intent-to-treat analysis of resource utilization in all patients with complete data sets (n = 78), hospitalization costs and crisis costs decreased significantly during the 12-month follow-up period (P = 0.003 and P = 0.009, respectively), and both outpatient and medication costs increased significantly (P = 0.035 and P < 0.001, respectively). Overall, the change in total annual resource utilization during the 12 months after initiation of olanzapine was not statistically significant (mean decrease per patient, $1,991; 95% CI, -$5,258 to $1,122). CONCLUSIONS: Initiation of olanzapine therapy was associated with favorable clinical outcomes in this population, particularly in patients with a formal diagnosis of thought disorder. Overall, there was a cost shift away from hospital and crisis costs toward medication and outpatient services costs. The decline in total resource utilization was not statistically significant, although it may be of practical importance. PMID- 12075949 TI - Length of hospital stay and cost of Staphylococcus and Streptococcus infections among hospitalized patients. AB - BACKGROUND: Staphylococcus (Staph) and Streptococcus (Strep) infections are important causes of morbidity and mortality worldwide. The economic burden of these infections is also significant, especially among hospitalized patients. OBJECTIVE: The aim of this study was to estimate length of hospital stay (LOS) and total payments for hospital admissions for patients with Staph or Strep infection as a first (primary) or second or higher (comorbid) diagnosis. METHODS: From the 1994-1997 MarketScan inpatient database, admissions with Staph (n = 2,042) or Strep (n = 1,401) infection (905 as primary and 2,538 as comorbid diagnosis) and 89,899 control admissions without a diagnosis of gram-positive infection were identified. Crude and category-specific mean LOS and anti-log mean total payments were compared between admissions with Staph or Strep infection and admissions without a diagnosis of any gram-positive infection within major diagnostic categories and principal surgical procedures (SPs). RESULTS: For admissions with Staph or Strep infection as first (primary) diagnosis (n = 905), the mean LOS was 4.68 days (95% CI, 4.44-4.93) and 4.78 days (95% CI, 4.35-5.26), respectively. The mean total payments were $6,445 (95% CI, $6,045-$6,870) and $6,821 (95% CI, $6,149-$7,566), respectively. In contrast, the average LOS and total payment for the control group were 2.99 days (95% CI, 2.98-3.01) and $6,325 (95% CI, $6,284-$6,365). For admissions with infection as the comorbid diagnosis (n = 2,538), mean LOS and total payment were 4 days longer and $6,000 higher for Staph infections and 1.2 days longer and $1,200 higher for Strep infections than the control group. Within each SP, LOS and total payments were substantially higher for patients with Staph and Strep infections. CONCLUSIONS: The results of this study indicate that infections with the pathogens Staph and Strep substantially increase LOS and total payments among hospitalized patients. PMID- 12075950 TI - Risedronate: a new oral bisphosphonate. PMID- 12075951 TI - Sterilising Siqveland. PMID- 12075952 TI - Setting the standard for dental health education. PMID- 12075954 TI - Emergency dental services. PMID- 12075953 TI - My view on fluoridation. PMID- 12075955 TI - Raising the ARF? PMID- 12075956 TI - Crowns and other extra-coronal restorations: preparations for full veneer crowns. AB - Preparations for full veneer crowns is the eighth in the series on crowns and other extra-coronal restorations. Whilst handpiece skills are important, many other factors combine to ensure provision of a satisfactory full veneer crown (also termed 'full coverage crown'). Our aim in writing this article is to consider the principles which influence crown preparation, seasoned with clinical advice our undergraduate and postgraduate students have found useful. PMID- 12075957 TI - Dental bodies corporate and their approach to customer care. AB - For many years the 27 bodies corporate registered with the General Dental Council were of little interest to most people in the UK dental profession, serving only as an anachronistic reminder of a bygone period. How times change. Although they still have only a small share of the dental market - with 4% of all dentists in the UK in early 1999 - they have expanded rapidly from a small base and are expected to continue to do so in the future. The reasons behind this growth are numerous and include such factors as: deregulation of the profession allowing dentists to advertise thus facilitating company branding; a general move away from NHS dentistry; a growing consumerism amongst the general public; precedents set by pharmacists and opticians; and, last but not least, the belief of venture capitalists amongst others that investment in dentistry will yield attractive returns. PMID- 12075958 TI - The long-term benefits of orthodontic treatment. AB - OBJECTIVE: This study aimed to evaluate post-treatment changes at least five years out of retention in a cohort of 100 consecutively started patients treated by a postgraduate student in a UK orthodontic department. DESIGN: Analysis of study models using recognised occlusal indices. Setting Patients initially treated at Newcastle Dental Hospital were recalled for record collection. MATERIALS AND METHODS: Study models and clinical photographs were taken for each patient attending for post-retention records. Weighted Peer Assessment Rating (PAR) Index and Index of Orthodontic Treatment Need (IOTN) scores were recorded for cases at the beginning, end and at least 5 years post-retention. Main outcome measures Post-treatment changes were evaluated using PAR Index and IOTN to evaluate extent and likelihood of change and to compare treatment modalities. RESULTS: Seventy-eight patients attended for post-retention records. Overall PAR reduction immediately post-treatment was 68.6% and at a mean of 6.5 years post retention was 55.5%. Dual arch fixed appliance treatments achieved the greatest reduction in PAR score, and maintained the reduction beyond the retention period better than other treatment modalities. Eighty-four per cent of the cohort still had some demonstrable improvement after a mean of 6.5 years post-retention. There was a residual need for treatment in 8% of cases on aesthetic grounds and in 24% of cases on dental health grounds. CONCLUSIONS: Occlusal deterioration after orthodontic treatment is almost universal. Dual arch fixed appliance treatment provides the best post-treatment and post-retention result. PMID- 12075960 TI - Focus Awards 2001. PMID- 12075961 TI - Propagation of limited-diffraction X-waves in dissipative media. AB - Diffractionless solutions of the wave equation in the form of X-waves have previously been obtained based on the inviscid form of the wave equation. A new general solution to the cylindrically symmetric wave equation for a medium with classical viscous losses is obtained. Particular solutions called dissipative Arcsin X-waves have been derived from this general solution. The properties of these waves are discussed for both infinite and finite size transducers and for different viscous liquids. To calculate the field produced by a finite transducer diameter, we have derived a dissipative form of the Rayleigh integral. PMID- 12075959 TI - Attitudes to biopsy procedures in general dental practice. AB - AIM: To investigate biopsy procedures in general dental practice. OBJECTIVES: To assess the views and attitudes of: specialists on the dental specialist surgical registers; dentists in general practice (GDPs) and patients undergoing biopsy procedures. METHOD: Questionnaires were sent to 98 oral and maxillofacial surgeons and surgical dentists, 335 general dental practitioners and 220 patients attending the Oral Medicine Clinic at the Dental Hospital, Manchester. Participation rates were 68 (74%), 227 (72%), and 158 (76%) respectively. RESULTS: Specialists: 47 (70%) would discourage dental practitioners undertaking biopsies. Concerns were a lack of skills and delays in referral; 20 (30%) considered GDPs should be able to perform simple biopsies for benign lesions. Dentists: 33 (15%) reported they had performed oral biopsies in the last two years; 136 (60%) felt they should be competent to biopsy benign lesions. Their main concerns were lack of practical skills and the risk of diagnostic error. PATIENTS: 112 (65%) worried about their biopsy result, 67 (39%) would feel anxious if their dentist did the biopsy, although 40 (23%) were anxious when biopsied in the oral medicine clinic. CONCLUSIONS: Both specialists on the dental surgical registers and GDPs feel there is a need for further training in biopsy technique for GDPs and better advertised and accessible pathology support. The current fee for biopsies may need upward revision. A main concern of patients is fear of an adverse biopsy report. Whilst patients are satisfied with specialist management any concerns were an insufficient reason for biopsy of a benign lesion not being undertaken in general practice. PMID- 12075962 TI - Doppler angle estimation using AR modeling. AB - The transit time spectrum broadening effect has long been explored for Doppler angle estimation. Given acoustic beam geometry, the Doppler angle can be derived based on the mean Doppler frequency and the Doppler bandwidth. Spectral estimators based on the fast Fourier transform (FFT) are typically used. One problem with this approach is that a long data acquisition time is required to achieve adequate spectral resolution, with typically 32-128 flow samples being needed. This makes the method unsuitable for real-time two-dimensional Doppler imaging. This paper proposes using an autoregressive (AR) model to obtain the Doppler spectrum using a small number (e.g., eight) of flow samples. The flow samples are properly selected, then extrapolated to ensure adequate spectral resolution. Because only a small number of samples are used, the data acquisition time is significantly reduced and real-time, two-dimensional Doppler angle estimation becomes feasible. The approach was evaluated using both simulated and experimental data. Flows with various degrees of velocity gradient were simulated, with the Doppler angle ranging from 20 degrees to 75 degrees. The results indicate that the AR method generally provided accurate Doppler bandwidth estimates. In addition, the AR method outperformed the FFT method at smaller Doppler angles. The experimental data for Doppler angles, ranging from 33 degrees to 72 degrees, showed that the AR method using only eight flow samples had an average estimation error of 3.6 degrees, which compares favorably to the average error of 4.7 degrees for the FFT method using 64 flow samples. Because accurate estimates can be obtained using a small number of flow samples, it is concluded that real-time, two-dimensional estimation of the Doppler angle over a wide range of angles is possible using the AR method. PMID- 12075963 TI - Clutter filters adapted to tissue motion in ultrasound color flow imaging. AB - The quality of ultrasound color flow images is highly dependent on sufficient attenuation of the clutter signals originating from stationary and slowly moving tissue. Without sufficient clutter rejection, the detection of low velocity blood flow will be poor, and the velocity estimates will have a large bias. In some situations, e.g., when imaging the coronary arteries or when the operator moves the probe in search for small vessels, there is considerable movement of tissue. It has been suggested that clutter rejection can be improved by mixing down the signal with an estimate of the mean frequency prior to high pass filtering. In this paper, we compare this algorithm with several other adaptive clutter filtering algorithms using both experimental data and simulations. We found that realistic accelerations of the tissue have a large effect on the clutter rejection. The best results were obtained by mixing down the signal with non constant phase increments estimated from the signal. This adapted the filter to a possibly accelerated tissue motion and produced a significant improvement in clutter rejection. PMID- 12075965 TI - Frequency dependence of speckle in continuous-wave ultrasound with implications for blood perfusion measurements. AB - Speckle in continuous wave (CW) Doppler has previously been found to cause large variations in detected Doppler power in blood perfusion measurements, where a large number of blood vessels are present in the sample volume. This artifact can be suppressed by using a number of simultaneously transmitted frequencies and averaging the detected signals. To optimize the strategy, statistical properties of speckle in CW ultrasound need to be known. This paper presents analysis of the frequency separation necessary to obtain independent values of the received power for CW ultrasound using a simplified mathematical model for insonation of a static, lossless, statistically homogeneous, weakly scattering medium. Specifically, the autocovariance function for received power is derived, which functionally is the square of the (deterministic) autocorrelation function of the effective sample volumes produced by the transducer pair for varying frequencies, at least if a delta correlated medium is assumed. A marginal broadening of the modeled autocovariance functions is expected for insonation of blood. The theory is applicable to any transducer aperture, but has been experimentally verified here with 5-MHz, 6.35-mm circular transducers using an agar phantom containing small, randomly dispersed glass particles. A similar experimental verification of a transducer used in multiple-frequency blood perfusion measurements shows that the model proposed in this paper is plausible for explaining the decorrelation between different channels in such a measurement. PMID- 12075964 TI - Decorrelation-based blood flow velocity estimation: effect of spread of flow velocity, linear flow velocity gradients, and parabolic flow. AB - In recent years, a new method to measure transverse blood flow, based on the decorrelation of the radio frequency (RF) signals has been developed. In this paper, we investigated the influence of nonuniform flow on the velocity estimation. The decorrelation characteristics of transverse blood flow using an intravascular ultrasound (IVUS) array catheter are studied by means of computer modeling. Blood was simulated as a collection of randomly located point scatterers; moving this scattering medium transversally across the acoustical beam represented flow. First-order statistics were evaluated, and the signal-to noise ratio from the signals were measured. The correlation coefficient method was used to present the results. Three velocity profiles were simulated: random spread of blood-flow velocity, linear blood-flow velocity gradient, and parabolic blood-flow. Radio frequency and envelope signals were used to calculate the decorrelation pattern. The results were compared to the mean decorrelation pattern for plug blood-flow. The RF signals decorrelation patterns were in good agreement with those obtained for plug blood flow. Envelope decorrelation patterns show a close agreement with the one for plug blood flow. For axial blood flow, there is a discrepancy between decorrelation patterns. The results presented here suggest that the decorrelation properties of an IVUS array catheter for measuring quantitative transverse blood flow probably will not be affected by different transverse blood-flow conditions. PMID- 12075967 TI - Estimation and compensation of ultrasonic wavefront distortion using a blind system identification method. AB - A common random input filter model is described for estimation and correction of wavefront aberration in ultrasonic b-scan imaging. In the model, aberration between the focus and the transducer elements is represented by the response of a linear filter bank to a common random signal. The response of each filter in the bank is found using a two-level extension of an existing subspace method for blind system identification. The receive waveforms are compensated using an inverse filter, and the transmit waveforms are predistorted using time reversal. To test the model, experiments were conducted using a two-dimensional array system to obtain echoes from a point reflector and from a random medium in each case through an aberrator. The aberrator is a phantom that mimics wavefront distortion produced by human abdominal wall, and the random medium is made to mimic ultrasonic characteristics of human liver. The results indicate the method can improve both the transmit and the receive focus and can outperform time-shift estimation and compensation as well as the method of backpropagation followed by timeshift estimation and compensation. PMID- 12075966 TI - In vivo droplet vaporization for occlusion therapy and phase aberration correction. AB - The objective was to determine whether a transpulmonary droplet emulsion (90%, <6 microm diameter) could be used to form large gas bubbles (>30 microm) temporarily in vivo. Such bubbles could occlude a targeted capillary bed when used in a large number density. Alternatively, for a very sparse population of droplets, the resulting gas bubbles could serve as point beacons for phase aberration corrections in ultrasonic imaging. Gas bubbles can be made in vivo by acoustic droplet vaporization (ADV) of injected, superheated, dodecafluoropentane droplets. Droplets vaporize in an acoustic field whose peak rarefactional pressure exceeds a well-defined threshold. In this new work, it has been found that intraarterial and intravenous injections can be used to introduce the emulsion into the blood stream for subsequent ADV (B- and M-mode on a clinical scanner) in situ. Intravenous administration results in a lower gas bubble yield, possibly because of filtering in the lung, dilution in the blood volume, or other circulatory effects. Results show that for occlusion purposes, a reduction in regional blood flow of 34% can be achieved. Individual point beacons with a +24 dB backscatter amplitude relative to white matter were created by intravenous injection and ADV. PMID- 12075968 TI - Piezoelectrically actuated flextensional micromachined ultrasound droplet ejectors. AB - This paper reports a variation on the design of the flextensional transducer for use in ejecting liquids. The transducer is constructed by depositing a piezoelectric thin film to a thin, edge-clamped, circular annular plate. By placing a fluid behind one face of a vibrating compound plate that has an orifice at its center, we achieve continuous or drop-on-demand ejection of the fluid. We present results of ejection of water and isopropanol. The ejector is harmless to sensitive fluids and can be used to eject fuels as well as chemical and biological samples. Micromachined two-dimensional array piezoelectrically actuated flextensional droplet ejectors were realized using planar silicon micromachining techniques. Typical resonant frequency of the micromachined device ranges from 400 kHz to 4.5 MHz. The ejection of water thru a 5-microm diameter orifice at 3.5 MHz was demonstrated by using the developed micromachined two dimensional array ejectors. PMID- 12075969 TI - Time of flight diffraction imaging for double-probe technique. AB - Due to rapid progress in microelectronics and computer technologies, the system evolving from analog to digital, and a programmable and flexible synthetic aperture focusing technique (SAFT) for the single-probe pulse-echo imaging technique of ultrasonic nondestructive testing (NDT) becomes feasible. The double probe reflection technique usually is used to detect the nonhorizontal flaws in the ultrasonic NDT. Because there is an offset between the transmitter and receiver, the position and size of the flaw cannot be directly read from the image. Therefore, a digital signal processing (DSP) imaging method is proposed to process the ultrasonic image obtained by double-probe reflection technique. In the imaging, the signal is redistributed on an ellipsoid with the transmitter and receiver positions as focuses, and the traveltime sum for the echo from the ellipsoid to the focuses as the traveltime of signal. After redistributing all the signals, the useful signals can be constructively added in some point in which the reflected point is; otherwise, the signals will be destructively added. Therefore, the image resolution of the flaw can be improved and the position and size of the flaw can be estimated directly from the processed image. Based on the experimental results, the steep flaw (45 degrees) cannot be detected by the pulse echo technique but can be detected by the double-probe method, and the double probe B-scan image of 30 degrees tilted crack is clearer than the pulse echo B scan image. However, the flaw image departs from its true position greatly. After processing, the steep flaw image can be moved to its true position. When the flaws are not greater than the probe largely, the sizes of the flaws are difficult to be discriminated in both pulse echo and double-probe B-scan images. In the processed double-probe B-scan image, the size of the flaws can be estimated successfully, and the images of the flaws are close to their true shape. PMID- 12075970 TI - AM noise impact on low level phase noise measurements. AB - The influence of the source AM noise in microwave residual phase noise experiments is investigated. The noise floor degradation problem, caused by the parasitic detection of this type of noise by an unperfectly balanced mixer, is solved thanks to a refinement of the quadrature condition. The parasitic noise contribution attributable to the AM to PM (phase modulation) conversion occurring in the device under test is minimized through the development of a dedicated microwave source featuring an AM noise level as low as -170 dBc/Hz at 10 kHz offset from a 3.5 GHz carrier. PMID- 12075971 TI - A simple optimally unbiased MA filter for timekeeping. AB - In this paper, we address the design of an optimally unbiased moving average (MA) filter for time error online estimating and synchronization in Global Positioning System (GPS)-based timekeeping. We proceed from the slowly changing nature of a time error generated by a local clock, assuming its function to be linear for N points in the average. Employing the inherent property of a simple MA to produce the lowest noise for the given averaging interval and using linear regression as a proper stochastic approximation to evaluate the bias, we find the optimal weighting coefficients. The designed optimally unbiased MA filter and its improved modification produce two times more noise than a simple MA when N >> 1. Both the numerical simulation and the filtering of the real GPS-based time error processes are provided and discussed in detail. PMID- 12075972 TI - Analysis of a rotating elastic beam with piezoelectric films as an angular rate sensor. AB - The flexural vibration of an elastic beam with surface-bonded piezoelectric films rotating about its axis is studied. One-dimensional equations governing the motion of the beam are developed, including the effects of Coriolis and centrifugal forces. The equations are used in the analysis of the flexural vibration of the beam under the excitation of an alternating electric voltage. Forced vibration solution is obtained. The beam can be used as a gyroscope for detecting the angular rate of the rotation. Voltage sensitivity and its dependence on various geometric and physical parameters are examined. PMID- 12075973 TI - SSBW to PSAW conversion in SAW devices using heavy mechanical loading. AB - The development of efficient computation tools based on mixed analytical and numerical calculation approaches allows precise descriptions and characterizations of surface acoustic waves (SAW) propagation, taking into account realistic electrical and mechanical boundary conditions. As an example, suppression of the leaky SAW, also called pseudo SAW (PSAW), attenuation has been predicted using such tools allowing to explain experimental occurrences for SAW devices on YX cut lithium niobate and lithium tantalate with thick aluminum strip gratings (6% < h/lambda < 10%). In this work, such a theoretical model is used to analyze the evolution of surface waves on standard YX lithium tantalate cuts versus aluminum strip height. It is shown that the surface skimming bulk wave (SSBW), which accompanies the pseudo SAW on such crystal orientations, may be trapped by the grating, exhibiting then a second pseudo SAW behavior when close to the Bragg condition. A device has been designed and fabricated to check these theoretical predictions. The experimental evidence of the existence of the phenomenon allows one to discuss its consequences on more classical devices built on (Y+36 degrees,X) LiTaO3 substrates. PMID- 12075974 TI - Metal strip reflectivity and NSPUDT orientations in langanite, langasite, and gallium phosphate. AB - New materials of the trigonal 32 class have received much attention recently, due to their quartz-like temperature behavior and the promise of higher electromechanical coupling coefficients. The magnitude and phase of the reflection coefficient of the metal strips that form the SAW transducers and reflector structures is a critical parameter that allows proper device designs and optimal material surface orientation. This paper describes an investigation of the magnitude and phase of the SAW metal strip reflectivity in some new materials and along new orientations. The materials are langanite, langasite, and gallium phosphate. The results are presented as contour plots of the magnitude and phase of the reflection coefficient. In addition, the phase velocity, temperature coefficient of delay, electromechanical coupling coefficient, and power flow angle are given, thus allowing proper orientation selection for SAW device designs. The results presented highlight the reflection coefficient calculations in the selection of natural single-phase unidirectional transducer orientations (NSPUDT) for these new materials. As a result of the present work, reflection coefficient magnitudes five times larger than those of the well-known quartz NSPUDT orientation (Euler angles: 0 degrees 132.75 degrees 25 degrees) can be obtained with the new materials, for orientations where the optimum NSPUDT reflection coefficient phase criterion is met. The new materials discussed here provide an opportunity to design shorter- and higher-bandwidth devices due to: the increased electromechanical coupling coefficient; the higher reflection coefficient; and the lower phase velocities; while maintaining the excellent temperature insensitivity characteristic of quartz. The orientation regions suggested in this paper for the new materials are thus very promising for low loss, high-performance consumer and communications SAW designs, such as NSPUDT filters, resonator-based filters, and other devices that can benefit from a high metal strip reflectivity. PMID- 12075975 TI - Comparison between ST-cut quartz 25 degrees and -60 degrees NSPUDT propagation directions. AB - This paper discusses directivity and other propagation properties of natural single phase unidirectional transducer (NSPUDT) propagation directions on ST-cut quartz. The work focuses on the comparison of surface acoustic wave (SAW) directivity and propagation properties between the ST-cut quartz -60 degrees, Euler angles: (0 degrees, 132.75 degrees, -60 degrees), and the ST-cut quartz 25 degrees, Euler angles: (0 degrees, 132.75 degrees, 25 degrees), NSPUDT propagation directions, including predicted and measured directivity responses for both propagation directions. The well-known SAW 25 degrees propagation direction is used for low loss, high performance SAW filter designs for consumer products and communications applications. The ST-cut quartz -60 degrees propagation direction has been predicted to have a reflection coefficient 2.5 times larger than ST-cut quartz 25 degrees. In addition the ST-cut quartz -60 degrees satisfied the NSPUDT 90 degrees reflection coefficient phase condition much more closely, resulting in an improved directivity response. For the delay line structures used in the experiments, the measured directivity is 10.1 dB for the -60 degrees propagation direction. For the same structures, the measured directivity along the 25 degrees propagation direction is about 5.0 dB. The experimental results given in this paper verify that indeed ST-cut quartz -60 degrees has a higher directivity than ST-cut quartz 25 degrees, confirming the theoretical predictions. In addition, this work compares other propagation properties for both directions, namely, phase velocities, electromechanical couplings, temperature coefficients of delay, power flow angles, and metallic strip reflection coefficient amplitudes and phases. PMID- 12075976 TI - High-accuracy standard specimens for the line-focus-beam ultrasonic material characterization system. AB - We prepared standard specimens for the line-focus-beam ultrasonic material characterization system to obtain absolute values of the propagation characteristics (phase velocity and attenuation) of leaky surface acoustic waves (LSAWs). The characterization system is very useful for evaluating and analyzing specimen surfaces. The calibration accuracy of these acoustic parameters depends on the accuracy of acoustical physical constants (elastic constants, piezoelectric constants, dielectric constants, and density) determined for standard specimens. In this paper, we developed substrates of nonpiezoelectric single crystals (viz., gadolinium gallium garnet [GGG], Si, and Ge) and an isotropic solid (synthetic silica [SiO2] glass) as standard specimens. These specimens can cover the phase velocity range of 2600 to 5100 m/s for Rayleigh type LSAWs. To determine the elastic constants with high accuracy, we measured velocities by the complex-mode measurement method and corrected diffraction effects. Measurements of bulk acoustic properties (bulk wave velocity and density) were conducted around 23 degrees C, and bulk wave velocities were obtained with an accuracy of within +/-0.004%. We clearly detected differences in acoustic properties by comparing the obtained results with the previously published values; the differences were considered to be due to differences of the specimens used. We also detected differences in acoustic properties among four SiO2 substrates produced by different manufacturers. PMID- 12075977 TI - Characterization of all the elastic, dielectric, and piezoelectric constants of uniaxially oriented poled PVDF films. AB - Polyvinylidene fluoride (PVDF), a piezoelectric material, has many useful applications, for example, as sensors, transducers, and surface acoustic wave (SAW) devices. Models of performance of these devices would be useful engineering tools. However, the benefit of the model is only as accurate as the material properties used in the model. The purpose of this investigation is to measure the elastic, dielectric and piezoelectric properties over a frequency range, including the imaginary part (loss) of these properties. Measurements are difficult because poled material is available as thin films, and not all quantities can be measured in that form. All components of the elastic stiffness, dielectric tensor, and electromechanical coupling tensor are needed in the models. The material studied here is uniaxially oriented poled PVDF that has orthorhombic mm2 symmetry. Presented are the frequency dependence of all nine complex elastic constants, three complex dielectric constants, and five complex piezoelectric constants. The PVDF was produced at Raytheon Research Division, Lexington, MA. Measurements were made on thin films and on stacked, cubical samples. The elastic constants c44D and C55D, the dielectric constants epsilon11T and epsilon22T, as well as the piezoelectric constants g15 and g24 reported here have not been published before. The values were determined by ultrasonic measurements using an impedance analyzer and a least square data-fitting technique. PMID- 12075978 TI - A low-cost, high-performance pulse generator for ultrasound imaging. PMID- 12075979 TI - Spatial separation of target and competitor cues enhances blocking of human causality judgements. AB - Three experiments were carried out. Each required subjects to make judgements about the causal status of cues following a two-stage blocking procedure. In Stage 1 a competitor cue was consistently paired with an outcome, and in Stage 2 the competitor continued to be paired with the outcome but was accompanied by a target cue. It was predicted that causal judgements for the target would be reduced by the presence of the competitor. In Experiments 1 and 2 the blocking procedure was implemented as a computer simulation of a card game during which subjects had to learn which cards produced the best payouts. The cues that subjects used to make their judgement were colours and symbols that appeared on the backs of the cards. When the target and competitor cues appeared on the same card blocking effects did not emerge, but when they appeared as part of different cards blocking effects were found. Thus, spatial separation of target and competitor cues appeared to facilitate blocking. Experiment 3 replicated the blocking result using spatially separated target and competitor cues. PMID- 12075980 TI - Higher-order retrospective revaluation in human causal learning. AB - Previous studies demonstrated that participants will retrospectively adjust their ratings about the relation between a target cue and an outcome on the basis of information about the causal status of a competing cue that was previously paired with the target cue. We demonstrate that such retrospective revaluation effects occur not only for target cues with which the competing cue was associated directly, but also for target cues that were associated indirectly with the competing cue. These second-order and third-order retrospective revaluation effects are compatible with certain implementations of the probabilistic contrast model and with a modified, extended comparator model, but cannot be explained on the basis of a revised Rescorla-Wagner model or a revised SOP model. PMID- 12075981 TI - Analysis of the perceptual learning effect in flavour aversion learning: evidence for stimulus differentiation. AB - Rats received exposure to two compound flavours, AX and BX, where A and B were sucrose and saline and X was acid. For group intermixed (1), exposure consisted of alternating trials with AX and BX; group blocked (B) received a block of AX trials and a separate block of BX trials. Experiment 1 showed that generalization to BX after conditioning with AX was less profound in group 1 than in group B. Separate examination of the elements of the compound showed that the source of this difference lay in the strength acquired by the X element. X acquired less strength in group 1 than in group B (Experiments 1 and 2), whereas for the A element (Experiments 3 and 4) the reverse pattern was obtained. These results support the proposal that the perceptual learning effect (restricted generalization from AX to BX in group 1) depends on a process that enhances the effectiveness of unique stimulus elements (A and B) and reduces that of common elements (such as X). PMID- 12075982 TI - Successive positive contrast in one-way avoidance learning. AB - The main finding of these experiments was a positive contrast effect in one-way avoidance learning. Experiment 1 showed that increasing safety time during one way avoidance training led to improved performance, surpassing that of a control group that had received the high reward (safe time) from the beginning of training. Experiment 2 showed that a similar positive contrast effect occurred when the time spent in the danger compartment before the onset of the warning signal was shortened. These results suggest that time spent in a safe context acts as a reinforcer of the avoidance response; however, its incentive value depends not only on its duration, but also on the length of the time spent in the danger compartment before the onset of the signal. Overall, results also suggest that the avoidance response is a mixture of flight (motivated by fear) and approach (to a safe place) behaviour. The specific weight of the flight or approach component may be a function of the time and the amount of activation of each emotional state (fear or relief) due to opponent homeostatic compensatory processes that occur in the danger and safe compartments during one-way avoidance learning. PMID- 12075983 TI - Contextual dependencies in a stimulus equivalence paradigm. AB - Two experiments with human subjects assessed contextual dependencies in a stimulus equivalence paradigm. Subjects learned to form two sets of stimuli in a matching-to-sample training procedure. Each set was presented against one of two different background colours, the contextual cues. At test, the influence of a context change-that is, presenting each set against the other context-was measured on baseline, symmetry, and equivalence trials. These three trial types reflect a difference in task complexity. It was predicted that the magnitude of context-dependent effects would be a positive function of task complexity. In Experiment 1, the context change was realized by switching the stimulus set at test while keeping the background colour constant. In Experiment 2, the stimulus set remained constant, and the background colour was switched. In both experiments, a change in context only resulted in an increase in response latency on equivalence trials; no effect was seen on symmetry and baseline trials. Results were discussed in the framework of switch costs, habituation to contextual stimuli, and a model based on Shea and Wright (1995) that explains the differential influence of a context switch on easy versus difficult tasks. PMID- 12075984 TI - A sequential intrastriatal dopaminergic graft strategy in the rodent model for Parkinson's disease: implications for graft survival and targeting. AB - Optimal placement of intrastriatal dopaminergic grafts is likely crucial to optimize clinical recovery in Parkinson's disease (PD). The target sites of dopaminergic grafts vary among clinical trials and may partially explain the variable results in clinical efficacy reported thus far. In this study we hypothesized that a subsequent dopaminergic graft may promote functional recovery following a suboptimal initial graft. To test this hypothesis, rats with unilateral 6-hydroxydopamine lesions of the right nigrostriatal pathway were randomly divided into three groups. The first group received 900,000 fetal nigral cells in the medial striatum only (n = 6). The second group received 900,000 cells in both the medial and lateral striatum simultaneously (1.8 million total; n = 8). The final group received a second graft of 900,000 cells in the lateral striatum 6 weeks following initial transplantation of a medial graft (n = 6). Amphetamine-induced circling behavior was significantly reduced in both simultaneous and sequential graft groups at 9 and 12 weeks following transplantation of the initial graft. However, no recovery was noted in the single medial graft group at those time points. Furthermore, increased survival of dopaminergic cells was observed in the lateral graft of sequentially grafted animals compared with the medial graft. We conclude that a well-positioned subsequent graft can restore function in animals with a suboptimal initial graft and that the initial graft may improve survival of the second graft. These results are further discussed in relation to their important clinical implication for neural transplantation in PD. PMID- 12075985 TI - Tauroursodeoxycholic acid improves the survival and function of nigral transplants in a rat model of Parkinson's disease. AB - There is accumulating evidence showing that the majority of cell death in neural grafts results from apoptosis when cells are implanted into the brain. Tauroursodeoxycholic acid (TUDCA), a taurine-conjugated hydrophilic bile acid, has been found to possess antiapoptotic properties. In the present study we have examined whether the supplementation of TUDCA to cell suspensions prior to transplantation can lead to enhanced survival of nigral grafts. We first conducted an in vitro study to examine the effects of TUDCA on the survival of dopamine neurons in serum-free conditions. The number of tyrosine hydroxylase (TH)-positive neurons in the TUDCA-treated cultures was significantly greater than that of control cultures 7 days in vitro. In addition, a terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) assay showed that the number of apoptotic cells in the TUDCA-treated cultures was dramatically smaller than that in the control cultures. In the transplantation study, a 50 microM concentration of TUDCA was added to the media when nigral tissue from Sprague-Dawley (SD) rats was trypsinized and dissociated. Two microliters of cell suspension containing TUDCA was then stereotaxically injected into the striatum of adult SD rats subjected to an extensive unilateral 6 hydroxydopamine lesion of the nigrastriatal dopamine pathway. At 2 weeks after transplantation, the rats that received a cell suspension with TUDCA exhibited a significant reduction in amphetamine-induced rotation scores when compared with pretransplantation value. There was a significant increase (approximately threefold) in the number of TH-positive cells in the neural grafts for the TUDCA treated group when compared with the controls 6 weeks postgrafting. The number of apoptotic cells was much smaller in the graft areas in the TUDCA-treated groups than in the control group 4 days after transplantation. These data demonstrate that pretreatment of the cell suspension with TUDCA can reduce apoptosis and increase the survival of grafted cells, resulting in an improvement of behavioral recovery. PMID- 12075986 TI - Striatal xenotransplantation of human retinal pigment epithelial cells attached to microcarriers in hemiparkinsonian rats ameliorates behavioral deficits without provoking a host immune response. AB - Attachment of donor cells to microcarriers has been reported to make them more tolerable for transplantation into the brain. Human retinal pigment epithelial (hRPE) cells have been previously reported to contain enzymes for the production of dopa. Therefore, we examined the host immune response and behavioral effects of xenotransplantation of hRPE cells attached to microcarriers (hRPE-M) into the striatum of unilateral dopamine-depleted rats. Thirty-four adult rats were lesioned with 6-OHDA injections into the medial forebrain bundle on the right side. After 5 weeks of testing for apomorphine-induced rotations (AIR), animals were randomized for right striatal surgery into the following four groups: hRPE-M (group 1), hRPE alone (group 2), microcarriers alone (group 3), or needle tract alone (group 4). Following surgery, animals were tested for AIR every 4 weeks for a period of 12-18 weeks and thereafter euthanized. There was a significant reduction in AIR scores posttransplantation in all groups of animals in the initial observation points at 4 weeks and 8 weeks. However, there was a gradual return to baseline scores in groups 2, 3, and 4 animals at 12 weeks and at 18 weeks only group 1 animals had statistically significant (p = 0.001, repeated measures ANOVA, means comparison, predetermined contrasts) reduction in AIR scores. Brain tissue from representative animals from each group was cut into 30 microm coronal sections, stained for cresyl violet, tyrosine hydroxylase (TH), and markers for host immune activation. Sections through the striatum from group 1 animals revealed microcarriers with attached cells resembling RPE cells. No evidence of transplanted hRPE cells could be detected in sections from group 2 animals while those from groups 3 and 4 animals showed microcarriers and a needle tract alone, respectively. There was no host TH-immunoreactive sprouting response in the striatum in any of the groups and the host immune response was minimal. These results suggest that intrastriatal hRPE-M xenotransplantation into rats is well tolerated without systemic immunosuppression and that such transplants may provide behavioral benefit for parkinsonism. PMID- 12075987 TI - Adeno-associated viral delivery of GDNF promotes recovery of dopaminergic phenotype following a unilateral 6-hydroxydopamine lesion. AB - Glial cell line-derived neurotrophic factor (GDNF) is a potent neurotrophic factor for dopamine neurons that has been proposed for use in the treatment of Parkinson's disease (PD). Previous studies using viral vectors to deliver GDNF in rodent models of PD have entailed administering the virus either prior to or immediately after neurotoxin-induced lesions, when the nigrostriatal pathway is largely intact, a paradigm that does not accurately reflect the clinical situation encountered with Parkinson's patients. In this study, recombinant adeno associated virus carrying the gene encoding GDNF (rAAV-GDNF) was administered to animals bearing a maximal lesion in the nigrostriatal system, more closely resembling fully developed PD. Rats were treated with 6-hydroxydopamine into the medial forebrain bundle and assessed by apomorphine-induced rotational behavior for 5 weeks prior to virus administration. Within 4 weeks of a single intrastriatal injection of rAAV-GDNF, unilaterally lesioned animals exhibited significant behavioral recovery, which correlated with increased expression of dopaminergic markers in the substantia nigra, the medial forebrain bundle, and the striatum. Our findings demonstrate that rAAV-GDNF is capable of rescuing adult dopaminergic neurons from near complete phenotypic loss following a neurotoxic lesion, effectively restoring a functional dopaminergic pathway and diminishing motoric deficits. These data provide further support for the therapeutic potential of rAAV-GDNF-based gene therapy in the treatment of PD. PMID- 12075988 TI - Upregulation of EphA receptor expression in the injured adult rat spinal cord. AB - After spinal cord injury (SCI), the inability of supraspinal neurons to regenerate or reform functional connections is likely due to proteins in the surrounding microenvironment restricting regeneration. EphAs are a family of receptor tyrosine kinases that are involved in axonal guidance during development. These receptors and their ligands, the Ephrins, act via repulsive mechanisms to guide growing axons towards their appropriate targets and allow for the correct developmental connections to be made. In the present study, we investigated whether EphA receptor expression changed after a thoracic contusion SCI. Our results indicate that several EphA molecules are upregulated after SCI. Using semiquantitative RT-PCR to investigate mRNA expression after SCI, we found that EphA3, A4, and A7 mRNAs were upregulated. EphA3, A4, A6, and A8 receptor immunoreactivity increased in the ventrolateral white matter (VWM) at the injury epicenter. EphA7 had the highest level of immunoreactivity in both control and injured rat spinal cord. EphA receptor expression in the white matter originated from glial cells as coexpression in both astrocytes and oligodendrocytes was observed. In contrast, gray matter expression was localized to neurons of the ventral gray matter (motor neurons) and dorsal horn. After SCI, specific EphA receptor subtypes are upregulated and these increases may create an environment that is unfavorable for neurite outgrowth and functional regeneration. PMID- 12075989 TI - Muscle reinnervation with delayed or immediate transplant of embryonic ventral spinal cord cells into adult rat peripheral nerve. AB - Muscle denervation is common in various neuromuscular diseases and after trauma. It induces skeletal muscle atrophy. Only muscle reinnervation leads to functional recovery. In previous studies, denervated adult rat muscles were rescued by transplantation of embryonic day 14-15 (E14-15) ventral spinal cord cells into a nearby peripheral nerve. In the present study, changes were made in the environment into which the cells were placed to test whether reinnervation was improved by: 1) prior nerve degeneration, induced by sciatic nerve transection 1 week before cell transplantation; 2) transplantation of 1 million versus 5 million cells; 3) addition of nerve growth factor (NGF) to the transplant. Ten weeks after cell transplantation, axons had grown from all of the transplants. The numbers of myelinated axons that regenerated into the tibial, medial (MG), and lateral gastrocnemius-soleus (LGS) nerves were similar across treatments. The mean diameters of large LGS axons (>6 microm) were significantly larger with nerve degeneration before transplantation. The mean diameters of MG and LGS axons were significantly larger with transplantation of 1 million versus 5 million cells. Silver-stained experimental and control lateral gastronemius (LG) muscles showed axons that terminated at motor end plates. Nodal and terminal sprouts were more common in reinnervated muscles (45-63% of all end plates) than in control muscles (10%). Electrical stimulation of the transplants induced weak contractions in 39 of 47 MG muscles (83%) and 33 of 46 LG muscles (72%) but at higher voltages than needed to excite control muscles. The threshold for MG contraction was lower with transplantation of 1 million cells, while LG thresholds were lower without NGF. The cross-sectional area of whole LG muscles was significantly larger with cell transplantation (immediate or delayed) than with media alone, but all of these muscle areas were reduced significantly compared with control muscle areas. These data suggest that delayed transplantation of fewer cells without NGF assists regeneration of larger diameter axons and prevents some muscle atrophy. PMID- 12075990 TI - T155g-immortalized kidney cells produce growth factors and reduce sequelae of cerebral ischemia. AB - Fetal rat kidney cells produce high levels of glial-derived neurotrophic factor (GDNF) and exert neuroprotective effects when transplanted into the brain in animal models of Parkinson's disease and stroke. The purpose of the present experiment was to produce kidney cell lines that secrete GDNF. Genes encoding two truncated N-terminal fragments of SV40 large T antigen, T155g and T155c, which does not code for small t antigen, were used. T155g was transduced into E17 cultured fetal Sprague-Dawley rat kidney cortex cells using a plasmid vector, and T155c was transduced with a plasmid and a retroviral vector. Sixteen clones were isolated from cultures transfected with the T155g-expressing plasmid. No cell lines were obtained with T155c. Four clones produced GDNF at physiological concentrations ranging from 55 to 93 pg/ml of medium. These four clones were transplanted into the ischemic core or penumbra of rats that had undergone middle cerebral artery occlusion (MCAO). Three of the four clones reduced the volume of infarction and the behavioral abnormalities normally resulting from MCAO. Blocking experiments with antibodies to GDNF and platelet-derived growth factor (PDGF) suggested that these growth factors contributed only minimally to the reduction in infarct volume and behavioral abnormality. These cell lines may be useful for intracerebral transplantation in animal models of brain injury, stroke, or Parkinson's disease. PMID- 12075991 TI - Human umbilical cord blood cells can be induced to express markers for neurons and glia. AB - Rare cells are present in human umbilical cord blood that do not express the hematopoietic marker CD45 and in culture do not produce cells of hematopoietic lineage. These umbilical cord multipotent stem cells (UC-MC) behave as multilineage progenitor cells (stem cells) and can be expanded in tissue culture. Exposure to basic fibroblast growth factor (bFGF) and human epidermal growth factor (hEGF) for a minimum of 7 days in culture induces expression of neural and glial markers. Western immunoblots demonstrate expression of both beta-tubulin III and glial fibrillary acidic protein (GFAP). Immunocytochemistry of the cells showed intense labeling to both compounds on the intracellular cytoskeleton. The oligodendrocyte cell surface marker galactocerebroside (Gal-C) was present on most cells. Many cells show dual labeling, expressing both neuronal and glial markers. PMID- 12075992 TI - Human umbilical cord blood cells express neural antigens after transplantation into the developing rat brain. AB - Recently, our laboratory began to characterize the mononuclear cells from human umbilical cord blood (HUCB) both in vitro and in vivo. These cryopreserved human cells are available in unlimited quantities and it is believed that they may represent a source of cells with possible therapeutic and practical value. Our previous molecular and immunocytochemical studies on cultured HUCB cells revealed their ability to respond to nerve growth factor (NGF) by increased expression of neural markers typical for nervous system-derived stem cells. In addition, the DNA microarray detected downregulation of several genes associated with development of blood cell lines. To further explore the survival and phenotypic properties of HUCB cells we transplanted them into the developing rat brain, which is known to provide a conducive environment for development of neural phenotypes. Prior to transplantation, HUCB cells were either cultured with DMEM and fetal bovine serum or were exposed to retinoic acid (RA) and nerve growth factor (NGF). Neonatal pups (1 day old) received unilateral injection of cell suspension into the anterior part of subventricular zone. One month after transplantation animals were perfused, their brains cryosectioned, and immunocytochemistry was performed for identification of neural phenotypes. Our results clearly demonstrated that approximately 20% of transplanted HUCB survived (without immunosuppression) within the neonatal brain. Additionally, double labeling with cell-type-specific markers revealed that some HUCB-derived cells (recognized by anti-human nuclei labeling) were immunopositive for glial fibrillary acidic protein (GFAP) and few donor cells expressed the neuronal marker TuJ1 (class III beta-tubulin). These findings suggest that at least some of the transplanted HUCB cells differentiated into cells with distinct glial or neuronal phenotypes after being exposed to instructive signals from the developing brain. PMID- 12075993 TI - Intravenous administration of human umbilical cord blood reduces neurological deficit in the rat after traumatic brain injury. AB - We measured the effect of treatment of traumatic brain injury (TBI) in the rat with human umbilical cord blood (HUCB) administered i.v.. HUCB cells were injected into the tail vein 24 h after TBI and the rats were sacrificed at day 28 after the treatment. The Rotarod test and the neurological severity score (NSS) were used to evaluate neurological function. The distribution of the donor cells in the brain, heart, lung, kidney, liver, spleen, bone marrow, and muscle were analyzed in recipient rats using immunohistochemical staining and laser confocal microscopy. HUCB cells injected i.v. significantly reduced motor and neurological deficits compared with control groups by day 28 after the treatment. The cells preferentially entered the brain and migrated into the parenchyma of the injured brain and expressed the neuronal markers, NeuN and MAP-2, and the astrocytic marker, GFAP. Some HUCB cells integrated into the vascular walls within the boundary zone of the injured area. Our data suggest that i.v. administration of HUCB may be useful in the treatment of TBI. PMID- 12075995 TI - TrkC overexpression enhances survival and migration of neural stem cell transplants in the rat spinal cord. AB - Although CNS axons have the capacity to regenerate after spinal cord injury when provided with a permissive substrate, the lack of appropriate synaptic target sites for regenerating fibers may limit restoration of spinal circuitry. Studies in our laboratory are focused on utilizing neural stem cells to provide new synaptic target sites for regenerating spinal axons following injury. As an initial step, rat neural precursor cells genetically engineered to overexpress the tyrosine kinase C (trkC) neurotrophin receptor were transplanted into the intact rat spinal cord to evaluate their survival and differentiation. Cells were either pretreated in vitro prior to transplantation with trkC ligand neurotrophin 3 (NT-3) to initiate differentiation or exposed to NT-3 in vivo following transplantation via gelfoam or Oxycel. Both treatments enhanced survival of trkC overexpressing stem cells to nearly 100%, in comparison with approximately 30-50% when either NT-3 or trkC was omitted. In addition, increased migration of trkC overexpressing cells throughout the spinal gray matter was noted, particularly following in vivo NT-3 exposure. The combined trkC expression and NT-3 treatment appeared to reduce astrocytic differentiation of transplanted neural precursors. Decreased cavitation and increased beta-tubulin fibers were noted in the vicinity of transplanted cells, although the majority of transplanted cells appeared to remain in an undifferentiated state. These findings suggest that genetically engineered neural stem cells in combination with neurotrophin treatment may be a useful addition to strategies for repair of spinal neurocircuitry following injury. PMID- 12075994 TI - Fibronectin promotes survival and migration of primary neural stem cells transplanted into the traumatically injured mouse brain. AB - Multipotential stem cells are an attractive choice for cell therapy after traumatic brain injury (TBI), as replacement of multiple cell types may be required for functional recovery. In the present study, neural stem cells (NSCs) derived from the germinal zone of E14.5 GFP-expressing mouse brains were cultured as neurospheres in FGF2-enhanced medium. When FGF2 was removed in vitro, NSCs expressed phenotypic markers for neurons. astrocytes, and oligodendrocytes and exhibited migratory behavior in the presence of adsorbed fibronectin (FN). NSCs (10(5) cells) were transplanted into mouse brains 1 week after a unilateral, controlled, cortical contusion (depth = 1 mm, velocity = 6 m/s, duration = 150 ms) (n = 19). NSCs were injected either directly into the injury cavity with or without an injectable FN-based scaffold [collagen I (CnI)/FN gel; n = 14] or into the striatum below the injury cavity (n = 5). At all time points examined (1 week to 3 months posttransplant), GFP+ cells were confined to the ipsilateral host brain tissue. At 1 week, cells injected into the injury cavity lined the injury penumbra while cells inserted directly into the striatum remained in or around the needle track. Striatal transplants had a lower number of surviving GFP+ cells relative to cavity injections at the 1 week time point (p < 0.01). At the longer survival times (3 weeks-3 months), 63-76% of transplanted cells migrated into the fimbria hippocampus regardless of injection site, perhaps due to cues from the degenerating hippocampus. Furthermore, cells injected into the cavity within a FN containing matrix showed increased survival and migration at 3 weeks (p < 0.05 for both) relative to injections of cells alone. These results suggest that FGF2 responsive NSCs present a promising approach for cellular therapy following trauma and that the transplant location and environment may play an important role in graft survival and integration. PMID- 12075996 TI - Evaluation of the effects of thiamethoxam on three species of African termite (Isoptera: Termitidae) crop pests. AB - Thiamethoxam formulated as ACTARA 25 WG has been tested on three species of higher African termites: Trinervitermes trinervius Rambur, Odontotermes smeathmani Fuller, and Amitermes evuncifer Silvestri, which are pests of the principal tropical food crops (e.g., rice, maize, sorghum, sugarcane). The doses used ranged from 0.03 to 10 ppm. The effective dose for these termites was 0.3 ppm, which resulted in 100% mortality within 2-8 d, depending on the species studied. Excluding O. smeathmani, which consumed the product, thiamethoxam proved to be an anti-feedant but not repellent to T. trinervius and A. evuncifer. For all species studied, mortality was not dependent on consumption of the products, which acts principally by contact. In O. smeathmani, thiamethoxam could be transmitted in the colony from contaminated individuals to healthy individuals. PMID- 12075997 TI - Field evaluation of potential of alarm pheromone compounds to enhance baits for control of grass-cutting ants (Hymenoptera: Formicidae). AB - Leaf-cutting ants are important economic pests of the Neotropics, and the most common method of control involves the use of insecticidal baits. Baits that are currently available exhibit low attractiveness to grass-cutting species, thus there is a need to develop improved baits. The potential for using alarm pheromone compounds to enhance the attractiveness and subsequent harvest of baits was examined for two economically important species of grass-cutting ant, Atta bisphaerica (Forel) and Atta capiguara (Goncalves). Compounds of the alarm pheromone were applied to rubber septa that were then sealed inside plastic sachets together with citrus pulp-based bait. The best candidate compound for bait enhancement was 4-methyl-3-heptanone. This compound significantly increased the attractiveness of bait sachets to both species. It also appeared to improve the discovery of nearby unenhanced sachets. However, 4-methyl-3-heptanone resulted in only a slight and non-significant improvement in bait harvest. Enhanced and unenhanced bait sachets were applied at a number of positions to obtain an improvement in harvest, but without success. The possible reasons for the lack of an enhancement of harvest and the potential for using alarm pheromone compounds as leaf-cutting ant bait enhancers are discussed. PMID- 12075998 TI - Population dynamics of Cacopsylla melanoneura (Homoptera: Psyllidae), a vector of apple proliferation phytoplasma in northwestern Italy. AB - Apple proliferation is a phytoplasma-associated disease transmitted by insects causing serious damage and economic losses to apple orchards. Investigations were carried out in 1999 and 2000 in northwestern Italy to identify the vector of apple proliferation and to study its population dynamics. Yellow sticky traps and beat tray samples revealed the presence of the psyllid Cacopsylla melanoneura (Forster) in eight apple orchards in the Aosta Valley. The species completes one generation per year; the overwintered psyllids colonized apple trees beginning in late January, whereas the springtime generation was observed beginning in early May. The offspring adults remained in apple orchards until the end of June, when they began to move onto other hosts. During 1999 and 2000, all apple trees present in the investigated orchards were visually checked to assess the fluctuation of disease symptoms. Polymerase chain reaction and restriction fragment-length polymorphism confirmed the presence of the apple proliferation phytoplasmas in both overwintering and offspring insects as well as in symptomatic apple plants. The ability of C. melanoneura to vector the disease was assessed by preliminary transmission trials. Overwintered psyllids, collected in the most affected orchards, caged on healthy apple test plants transmitted apple proliferation phytoplasmas. PMID- 12075999 TI - Influence of alfalfa cultivar on suitability of Acyrthosiphon kondoi (Homoptera: Aphididae) for survival and development of hippodamia convergens and Coccinella septempunctata (Coleoptera: Coccinellidae). AB - Hippodamia convergens Guerin-Meneville and Coccinella septempunctata L. (Coleoptera: Coccinellidae) larvae were supplied daily with 1, 2, 4, or 16 mg of Acyrthosiphon kondoi Shinji (Homoptera: Aphididae) reared on one of two susceptible ('OK08' or 'CUF-101') or one resistant ('54H55') alfalfa cultivar (IMedicago sativa L.) . Hippodamia convergens survived to the adult stage when supplied with > or = 1 mg of A. kondoi per day from both susceptible and aphid resistant cultivars, whereas C. septempunctata required > or = 2 mg of A. kondoi per day (from each cultivar) for survival to the adult stage. For both H. convergens and C. septempunctata, no consistent differences in survivorship or developmental times were observed between predator larvae supplied with increasing daily levels of A. kondoi from susceptible (OKO8 orCUF-101) versus resistant (54H55) cultivars. Additionally, alfalfa cultivar had no indirect influence on adult weight of H. convergens or C. septempunctata. Results from our study suggest that the resistant alfalfa cultivar (54H55) would have little to no effect on the nutritional value of A. kondoi for ladybeetle predators. PMID- 12076000 TI - Impact of Thelohania solenopsae (Microsporidia: Thelohaniidae) on polygyne colonies of red imported fire ants (Hymenoptera: Formicidae). AB - Three studies were conducted to assess the effects of the entomopathogen Thelohania solenopsae on polygynous, red imported fire ant, Solenopsis invicta Buren, colonies. A total of 57 of 122 queens (46.7%) from nine, field-collected, polygyne, S. invicta colonies, was infected with T. solenopsae. Infection rate of queens for each colony ranged from 25 to 75%. Laboratory colonies of polygyne S. invicta, with three to 12 queens, were inoculated and infected with T. solenopsae. Brood levels in all infected colonies declined to 0 after 26-52 wk. Brood did not reappear in any of the colonies after 3-11 wk, even though in two of the eight infected colonies, five fertile queens that were uninfected were recovered. Thus, polygyne, S. invicta colonies infected with T solenopsae, which were confined and isolated under laboratory conditions, did not recover. Field plots that contained polygynous S. invicta colonies, which were infected with T solenopsae, were monitored over a 2-yr period. Infection rates increased during the study and reached a maximum of 9.3%. Fire ant nest density and colony sizes fluctuated over time, with maximum reduction of 63% per plot. In general, fire ant reductions were attributed to smaller colony sizes. T. solenopsae infections in polygynous S. invicta can result in a slow colony decline and death. Under field conditions, the prolonged colony death may mask the impact of T. solenopsae by allowing for concurrent reinfestations. PMID- 12076001 TI - Development, survival, body weight, longevity, and reproductive potential of Oemena hirta (Coleoptera: Cerambycidae) under different rearing conditions. AB - Oemona hirta (F.) is a New Zealand native longicorn beetle, whose larvae bore into the wood of branches and stems of living trees and vines, causing serious damage. To explore effective methods for maintaining laboratory colonies and biology of immature stages of this species we evaluated four laboratory rearing methods with both natural and artificial diets and compared biological parameters of laboratory colonies with those of field-collected insects. On an artificial diet, approximately 40% of neonate and 70% of autumn- and 11% of winter-collected larvae reached adulthood. Neonate larvae could not complete their development in cut poplar (Populus nigra variety italica Koehne) twigs; however, when twigs were standing in water >46% of neonate larvae survived to adulthood. Mean larval development time ranged from approximately150 to almost 300 d, depending on rearing methods. Mean pupal stage ranged from 15 to 19 d. Adult females were significantly heavier than males. Although adult females from field-collected twigs and reared on the artificial diet had similar mean body weight, which was significantly greater than that of other rearing colonies, the potential fecundity was the highest in the former (83.1 +/- 29.4 eggs) and the lowest in the latter (33.5 +/- 9.1 eggs). In all laboratory-reared and field-collected insects, the total number of eggs produced (eggs laid + eggs that remained in dead female body) by females was positively correlated with their body weight. When larvae fed on the artificial diet, there was no positive correlation between the number of eggs successfully laid and female body weight. However, when larvae lived on natural food, a positive correlation was found. In laboratory colonies, mean longevity of females (36 to 52 d) was slightly greater than that of males (30 to 50 d) without significant difference between sexes but in the adults from field-collected twigs, males (52 d) lived significantly longer than females (33 d). In terms of time, labor, and the number of resulting adults, collecting larvae in the field in autumn and then transferring them onto artificial diet is the most effective method for maintaining a laboratory colony. PMID- 12076003 TI - Evaluation of tebufenozide carry-over and residual effects on spruce budworm (Lepidoptera: Tortricidae). AB - Laboratory and field studies investigated carry-over effects of tebufenozide on spruce budworm, Choristoneura fumiferana (Clemens). In the laboratory, third and fourth instars were fed either sublethal doses of tebufenozide (10 ppm) or water onAbies balsamea (L.) Mill. needles, reared to adulthood and allowed to oviposit on laying surfaces 1 or 14 d after being sprayed with water or tebufenozide concentrations of 17.5, 35.0, and 70.0 g/liter. Percentage adult emergence and sex ratio were not affected by larval ingestion of the tebufenozide. Also, the mean number of eggs laid on untreated wax paper by moths reared from tebufenozide treated larvae was similar to the controls. Hence, tebufenozide did not inhibit carry-over effects on treated larvae. Oviposition on tebufenozide-treated wax paper by moths reared from untreated larvae was affected by both the substrate concentration and the age of the treatment residue. When offered treated and untreated laying surfaces simultaneously, C. fumiferana did not show a preference. However, significantly fewer eggs were laid on both laying surfaces by fewer females than when tebufenozide was absent. Residual tebufenozide on wax paper did not affect egg hatch but topical applications were toxic to eggs. Field studies appear to corroborate laboratory results and suggest that although the ingestion of tebufenozide by larval spruce budworm might not impair adult reproduction, the insecticide's presence in the environment could inhibit oviposition. This inhibition was considered to be a primary factor in tebufenozide's multi-year effects against spruce budworm populations. PMID- 12076002 TI - Effects of row spacing and plant density on corn rootworm (Coleoptera: Chrysomelidae) emergence and damage potential to corn. AB - Planting corn, Zea mays L., in row spacings less than the conventional width of 76 cm has been shown to increase grain yields. This study was conducted to determine if row spacing and plant density affected corn rootworm, Diabrotica virgifera virgifera LeConte and D. barberi Smith & Lawrence, adult emergence, larval injury to the roots, and plant tolerance to injury. Field experiments were conducted at Ames and Nashua, IA, in 1998, 1999, and 2000. Treatments were row spacings of 38 and 76 cm, and plant populations of 64,500 and 79,600 plants per hectare. Adult emergence was 31% greater in 38 cm compared with 76-cm rows. However, root injury was not significantly different between row spacings or plant populations. Row spacing alone did not significantly influence tolerance to injury, measured as root size and the amount of root regrowth. However, at one environment where precipitation was low, plants in 38-cm rows produced 25% more regrowth compared with plants in 76-cm rows. Root dry weight and regrowth were suppressed by 16 and 32%, respectively, at the high plant population. Although lodging was 51% lower in the 38-cm rows compared with the 76-cm rows, grain yields were not significantly different between row spacings. Reducing the row spacing of field corn from 76-38 cm should not increase the potential for injury from corn rootworm larvae. PMID- 12076004 TI - Effects of artificial and western spruce budworm (Lepidoptera: Tortricidae) defoliation on growth and biomass allocation of Douglas-fir seedlings. AB - Artificial defoliation has been used commonly to simulate defoliation by insect herbivores in experiments, in spite of the fact that obvious differences exist between clipping foliage and natural defoliation due to insect feeding. We used a greenhouse experiment to compare the effects of artificial and western spruce budworm (Choristoneura occidentalis Freeman) defoliation on the growth and biomass allocation of 3-yr old half-sib seedlings from mature Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco variety glauca] trees that showed phenotypic resistance versus susceptibility to budworm defoliation in the forest. Artificial clipping of buds mimicked the effects of budworm feeding on total seedling biomass when 50% of the terminal buds were damaged. However, artificial defoliation decreased seedling height, relative growth rate of height, and shoot: root ratio more than budworm defoliation, whereas budworm defoliation decreased stem diameter relative growth rate more than artificial defoliation. Half-sib seedling progeny from resistant maternal tree phenotypes had greater height, diameter, biomass, and shoot: root ratio than seedlings from susceptible phenotypes. We concluded that careful artificial defoliation could generally simulate effects of budworm defoliation on total biomass of Douglas-fir seedlings, but that the two defoliation types did not have equal effects on biomass allocation between shoot and root. Further, an inherently higher growth rate and a greater allocation of biomass to shoot versus root are associated with resistance of Douglas-fir trees to western spruce budworm defoliation. PMID- 12076005 TI - Watermelon seedling growth and mortality as affected by Anasa tristis (Heteroptera: Coreidae). AB - Adult squash bugs, Anasa tristis (De Geer), were confined on seedling watermelon plants at densities of zero, one, two, and four per plant. Squash bugs were allowed to feed on the plants until plants died or reached 30 cm in height. Number of leaves and length of plant vine were recorded at 2- or 3-d intervals. Seedling foliage, stems, and roots were harvested and dried after plants reached 30 cm in height. Growth of seedlings was regressed on number of squash bugs and results indicated that an increasing density of squash bugs feeding on seedlings resulted in a significant reduction in plant growth. Additionally, increased density of squash bugs resulted in reduced weight of foliage and root dry biomass. Seedling mortality increased as the density of squash bugs increased. PMID- 12076006 TI - Susceptibility of black soldier fly (Diptera: Stratiomyidae) larvae and adults to four insecticides. AB - Dosage-mortality regressions were determined for black soldier fly, Hermetia illucens (L.), larvae fed cyromazine or pyriproxifen treated media. Cyromazine LC50 for larvae dying before becoming prepupae ranged from 0.25 to 0.28 ppm with dosage-mortality regression slopes between 5.79 and 12.04. Cyromazine LC50s for larvae dying before emergence ranged from 0.13 to 0.19 ppm with dosage-mortality regression slopes between 3.94 and 7.69. Pyriproxifen dosage-mortality regressions were not generated for larvae failing to become prepupae since <32% mortality was recorded at the highest concentration of 1,857 ppm. LC50s for larvae failing to become adults ranged from 0.10 to 0.12 ppm with dosage mortality-regression slopes between 1.67 and 2.32. Lambda-cyhalothrin and permethrin dosage-mortality regressions were determined for wild adult black soldier flies and house flies, Musca domestica L., and for susceptible house flies. Our results indicate that the wild house fly, unlike the black soldier fly, population was highly resistant to each of these pyrethroids. Regression slopes for black soldier flies exposed to lambda-cyhalothrin were twice as steep as those determined for the wild house fly strain. Accordingly, LC50s for the black soldier fly and susceptible house fly were 10- to 30-fold lower than those determined for wild house flies. The differential sensitivity between wild black soldier flies and house flies might be due to behavioral differences. Adult house flies usually remain in animal facilities with the possibility of every adult receiving pesticide exposure, while black soldier fly adults are typically present only during emergence and oviposition thereby limiting their exposure. PMID- 12076008 TI - Piperidine alkaloids in sitka spruce with varying levels of resistance to white pine weevil (Coleoptera: Curculionidae). AB - Our objective was to evaluate piperidine alkaloids as potential resistance factors in Sitka spruce, Picea sitchensis (Bong.) Carr, at risk to attack by white pine weevils, Pissodes strobi (Peck). We sampled 72 seedlings in each of two replicated field trials in the Oregon Coast Range. The seedlings were grown from open-pollinated seeds of putatively "resistant" or "susceptible" off-site parental sources. Alkaloid concentrations in bark and foliage were measured in previously unattacked trees at the time of weevil host selection. Leader mortality was evaluated in the fall to gauge actual resistance in the sample trees. Five families had < or = 25% topkill and seven sustained >50% topkill. Alkaloid concentrations differed significantly among families, but the major alkaloids did not appear to be functionally linked with topkill or useful indicators of resistance. However, our study design did not address all potential resistance mechanisms. Therefore, before concluding that Sitka spruce alkaloids have no influence on white pine weevils, complementary laboratory and field experiments are needed. PMID- 12076007 TI - Proteins as active compounds involved in insecticidal activity of mushroom fruitbodies. AB - Many mushrooms are toxic to insects. To identify the chemicals involved in insecticidal activity, the toxicity of 14 species has been studied for water solubility, thermolability, and dialysis. The data strongly suggest that proteins are responsible for most of the insecticidal activity of mushroom fruitbodies and may be a source of genes available for plant protection against insects. Among proteins, lectins and hemolysins were good insecticide candidates because the toxicities were not affected by protease. PMID- 12076009 TI - Survival of kansas dipel-resistant European corn borer (Lepidoptera: Crambidae) on Bt and non-Bt corn hybrids. AB - The Kansas Dipel-resistant and susceptible European corn borer, Ostrinia nubilalis (Hubner), were evaluated in the greenhouse on different Bt transgenic events expressed in corn hybrids. There were important differences in the resistance offered by the different Bt event corn hybrids. Hybrid comparison tests indicate that these Dipel-resistant first-instar European corn borer were not able to survive to adulthood on whorl-stage MON810, Bt11, or 176 Bt event corn plants. Third instars did not survive to adulthood on whorl-stage MON810 or Bt11 event corn plants but a small number of fifth instars were found on whorl stage DBT418 plants infested with Dipel-resistant larvae. First and third instars of these Dipel-resistant European corn borers caused more leaf-feeding damage and more tunneling on whorl-stage Bt-corn plants than did the Dipel-susceptible European corn borers. However, in the single Bt corn hybrid test, there was no survival of the Dipel-resistant European corn borers on DK580BtX or MAX454 Bt plants 35 to 42 d after they had been infested with first instars. These results demonstrate that the current Kansas selection of Dipel-resistant European corn borer strain cannot establish reproducing populations in the tested Bt corn lines and hybrids. PMID- 12076011 TI - Corn earworm, Helicoverpa zea (Lepidoptera: Noctuidae) and other insect associated resistance in the maize inbred Tex6. AB - A 2-yr field and laboratory study investigated insect resistance of the maize, Zea mays L., inbred Tex6, which has previously demonstrated resistance to Aspergillus ear rot and aflatoxin production, relative to susceptible inbred B73. Field studies indicated significantly greater resistance to insect feeding of V4 V8 growth stage Tex6 plants compared with B73 plants in both years, primarily to flea beetles (Chaetonema spp.). Field studies of natural (1999) and artificial (2000) infestations of corn earworms, Helicoverpa zea (Boddie), indicated much lower levels of kernel damage at milk stage (approximately three-fold) and smaller surviving larvae (approximately three-fold) in Tex6 compared with B73 ears. At harvest similar trends in reduction of numbers of damaged kernels per ear, as well as incidence and numbers of kernels per ear symptomatically infected by Fusarium spp. were noted. Laboratory studies indicated little difference in mortality or survivor weight of caterpillars or sap beetle adults caged with milk stage kernels of the two inbreds. However, assays with silks indicated significantly greater mortality of H. zea in both 1999 and 2000, and European corn borer, Ostrinia nubilalis (Hubner) in 1999 (only year tested) when fed Tex6 silks compared with B73 silks. Pollinated Tex6 silks were generally darker colored and more toxic than unpollinated silks. Thus, it is possible that commercially usable inbreds with resistance to insects, which also contribute to the mycotoxin problem through vectoring and damage, could be produced using Tex6 as a source. PMID- 12076010 TI - Effects of transgenic bt corn on growth and development of the stalk borer Papaipema nebris (Lepidoptera: Noctuidae). AB - This study assessed the efficacy of two different genetic events, event Bt 11 (CrylAb) and event CBH351 (Cry 9C), in Bt corn against two instar classes of the stalk borer Papaipema nebris across three different plant stages (V1, V3, and V5) of corn, Zea mays. Class A includes instars 1 and 2, and class B includes instars 3 and 4. Stalk borer response and development over time were measured, and the data from 1999 and 2000 show that the Bt corn does have some effect on the feeding and development of P. nebris. Injury to the corn plant was reduced, although not eliminated. Stalk borer larvae caused significantly (P = 0.0001) more injury to the non-Bt plants than to the Bt plants over time. Growth and development of the larvae were slowed and mortality was higher for Bt corn than for non-Bt corn. These data suggest that planting Bt corn may benefit growers by reducing, but not eliminating, stalk borer infestations and subsequent plant injury. PMID- 12076012 TI - Binary insecticidal crystal protein from Bacillus thuringiensis, strain PS149B1: effects of individual protein components and mixtures in laboratory bioassays. AB - A family of novel binary insecticidal crystal proteins, with activity against western corn rootworm, Diabrotica virgifera virgifera LeConte, was identified from Bacillus thuringiensis Berliner. A binary insecticidal crystal protein (bICP) from B. thuringiensis strain PS149B1 is composed of a 14-kDa protein (Cry34Abl) and a 44-kDaprotein (Cry35Ab1). These proteins have been co-expressed in transgenic maize plants, Zea mays L., and effectively control western corn rootworm larvae under field conditions. Laboratory experiments were conducted to better understand the contribution of each component protein to the in vivo activity of the bICP. The 14-kDa protein is active alone against southern corn rootworm, Diabrotica undecimpunctata howardi Barber, and was synergized by the 44 kDa protein. In mixtures, the concentration of the 14-kDa protein had a greater impact on efficacy than the 44-kDa component. Although both proteins are clearly required for maximal insecticidal activity, laboratory results did not support the formation of a stable, fixed-ratio complex of the two component proteins. PMID- 12076013 TI - Effectiveness of spinosad on four classes of wheat against five stored-product insects. AB - Spinosad is a commercial reduced-risk pesticide that is naturally derived. Spinosad's performance was evaluated on four classes of wheat (hard red winter, hard red spring, soft red winter, and durum wheats) against adults of the lesser grain borer, Rhyzopertha dominica (F.); rice weevil, Sitophilus oryzae (L.); sawtoothed grain beetle, Oryzaephilus surinamensis (L.); red flour beetle, Tribolium castaneum (Herbst); and larvae of the Indianmeal moth, Plodia interpunctella (Hubner). Beetle adults (25) or P. interpunctella eggs (50) were exposed to untreated wheat and wheat treated with spinosad at 0.1 and 1 mg (AI)/kg of grain. On all untreated wheat classes, adult beetle mortality ranged from 0 to 6%, and P. interpunctella larval mortality ranged from 10 to 19%. The effects of spinosad on R. dominica and P. interpunctella were consistent across all wheat classes. Spinosad killed all exposed R. dominica adults and significantly suppressed progeny production (84-100%) and kernel damage (66-100%) at both rates compared with untreated wheat. Spinosad was extremely effective against P. interpunctella on all wheat classes at 1 mg/kg, based on larval mortality (97.6-99.6%), suppression of egg-to-adult emergence (93-100%), and kernel damage (95-100%), relative to similar effects on untreated wheats. The effects of spinosad on S. oryzae varied among wheat classes and between spinosad rates. Spinosad was effective against S. oryzae, O. surinamensis and T. castaneun only on durum wheat at 1 mg/kg. Our results suggest spinosad to be a potential grain protectant for R. dominica and P. interpunctella management in stored wheat. PMID- 12076014 TI - Cone-dominated ERG luminance-response function: the Photopic Hill revisited. AB - PURPOSE: In response to progressively brighter stimuli, the b-wave of the photopic ERG gradually augments in amplitude, reaches a plateau for a narrow range of intensities and then rapidly decreases with further increments in the luminance of the flash. This unique luminance-response function was originally introduced as the Photopic Hill. The purpose of this study was to further characterize this unique feature of the cone ERG, investigate if it was only limited to b-wave measurements and if it could be obtained under different photopic background luminances. METHODS: Photopic ERGs and oscillatory potentials were generated in response to flashes of light ranging from 0.5 to 16 cd m(-2) s in intensity and presented against photopic backgrounds varying from 18 to 525 cd m(-2) in luminance. RESULTS: All but the brightest background yielded a clear Photopic Hill like luminance-response function which could only be evidenced with the b-wave, the i-wave and OP4 amplitude measurements. Interestingly, the maximal amplitude reached remained almost identical irrespective of the background luminance. CONCLUSIONS: Our results suggest that the retinal mechanisms at the origin of the Photopic Hill effect could represent a voltage limitation mechanism, intimately tied to the OFF pathway. The latter would however be intrinsic to the cone system only and not to the entire retinal network since significantly higher peak amplitudes are reached with dark adaptation. PMID- 12076015 TI - An expanded nystagmus acuity function: intra- and intersubject prediction of best corrected visual acuity. AB - The Nystagmus Acuity Function (NAF) provides an objective measurement of the foveation characteristics of nystagmus waveforms and an assessment of potential visual acuity for subjects with congenital (CN) or latent/manifest latent (LMLN) nystagmus. It is based on the subjects' ability to maintain fixation within a physiologically based 'foveation window' of +/- 0.5 degrees and +/- 4.0 degrees/s. However, some subjects are incapable of controlling fixation well enough to remain within this window with duration sufficient for good foveation. To obtain a measure of the CN waveforms of these individuals, we are proposing an eXpanded Nystagmus Acuity Function (NAFX) that relaxes either the position limit, the velocity limit, or both. Data used in this study comes from 11 human subjects with CN (10 idiopathic and 1 with achiasma) and a Belgian sheepdog with achiasma. Visual acuity was tested with a standard Snellen chart and eye movements recorded with infrared oculography or scleral search coil. For the NAFX to be useful, it must not only be applicable for subjects who cannot maintain fixation within the standard limits of the NAF, but also must yield results equivalent to those obtained with the NAF when testing subjects who are capable of maintaining good fixation control. For the latter subjects, the amount of time when position and velocity fell within the expanded limits was measured, the standard deviations of the position and velocity during these times were calculated, and a tau-surface for the exponential function was generated to guarantee the equivalence between the NAF and the NAFX. We developed an automated NAFX equivalent to the original NAF. We demonstrated that equivalence in 10 subjects and the use of the NAFX on two additional (1 human and 1 canine) subjects who were incapable of maintaining fixation within the standard position and velocity limits. We demonstrated the effects of surgery and related the results to visual acuity. We found the results to be comparable to those seen when applying the NAF to subjects who had good fixation control. The NAFX can be determined for CN and LMLN subjects with poor control of fixation by extending the standard NAF position and/or velocity limits for foveation. The resulting function can be used along with the longest foveation domain (derived from the NAFX to measure breadth of a high-NAFX region) to identify the gaze or convergence angles with the best waveform and to predict the best-possible visual acuity that could be achieved with the reduction of their nystagmus. PMID- 12076016 TI - Recognition of small stimulus screen masks using the multifocal ERG. AB - To evaluate the ability of the multifocal ERG (mfERG) to detect small defects in the stimulus array was the objective of this paper. Seven normal subjects had mfERGs recorded with a VERIS system. Stimulus arrays composed of 61, 103 or 241 hexagons were covered in part by small masks of different light transmittance properties. Only masks that covered at least one-half of a single 103 hexagon stimulus cell caused a significant reduction in signal. Different-shaped masks of about 5 degrees diameter were detectable using a 61-hexagon array only when they fully covered a stimulus cell. Detection was better, but marginal for some of the masks, with the 103 hexagon array. The 241 hexagon array showed sharp defects for all masks. Masking the stimulus screen is not equivalent to having a pathologic scotoma, but it demonstrates the greatest possible spatial sensitivity of the mfERG system. Thus, the mfERG appears to be able to detect small retinal lesions if they reduce local retinal function by at least 50% and correspond to at least half the area of one stimulus hexagon. Scotomas 5 degrees or smaller would be best detected using a fine (241 hexagon) stimulus array. With coarser stimulus arrays (e.g. 103 or 61 hexagons), the effect of a small scotoma depends on its location relative to the stimulus cells. These issues should be considered when selecting mfERG recording conditions. PMID- 12076018 TI - Quantifying the benefits of additional channels of multifocal VEP recording. AB - For some individuals and for some locations, multifocal visual evoked potentials (mfVEP) may be too small or appear 'too noisy' to be reliably measured. By adding electrodes, especially electrodes placed lateral to the midline, and by recording with multiple channels, the amplitude of the signal can be increased in some field locations. However, the addition of electrodes involves certain costs; the set-up time is longer and the data analysis more time consuming and complex. The objective of this study was to assess the benefits of adding electrodes by quantifying these benefits using a signal-to-noise measure. In addition to the typical midline placement of electrodes, two electrodes were placed 1 cm above and 4 cm lateral to the inion on each side. This allowed for 3 channels of recording and 3 additional, derived channels. The mfVEPs were recorded with a 60 sector, pattern-reversing display presented to one eye. Two 7 min records were obtained from 14 individuals with no known visual problems. The two records were averaged and a signal-to-noise (SNR) measure was obtained for every response from all 6 channels. For each sector of the display and each subject, the benefits of additional electrodes were quantified by comparing the SNR from the traditional midline channel to the best SNR from amongst the 6 channels. The number of responses exceeding any given criterion SNR value was increased with the additional channels. For example, 79% of the responses for the typical midline channel exceeded a SNR of 0.6 (a false positive rate of about 2.5%) and this increased to 93% when the best SNR value was used. As expected, summing the mfVEP responses from contiguous sectors also increased the SNR values. Additional electrodes and multiple channels of recording substantially improve the quality of the mfVEP records and the SNR measure provides a useful metric for assessing these benefits. PMID- 12076017 TI - A signal-to-noise analysis of multifocal VEP responses: an objective definition for poor records. AB - Sixty local VEP records, called the multifocal VEP (mfVEP), can be obtained over a wide retinal area. From subject-to-subject, from day-to-day, and from location to-location, these records can vary in quality presenting a challenge to quantitative analyses. Here three procedures are described for specifying the quality of mfVEP recordings in terms of signal-to-noise ratios. Monocular mfVEPs were recorded in two, 7-min runs. A '2-run signal-to-noise ratio' (2rSNR) was obtained as [RMS(RunA+RunB)]/[RMS(RunA-RunB)]-1, where RMS is the root-mean square amplitude of the response over the period from 45 to 150 ms (signal window). Two 'noise-window signal-to-noise ratios' were obtained with the same numerator as the 2rSNR but with the denominators based upon the RMS of a signal free window from 325 to 430 ms. In one case, inSNR, the denominator was the RMS of the record's noise window and in the other case, mnSNR, the denominator was the mean of the RMS amplitudes of all the signal-free noise windows for the subject. The SNRs were related to false-positive rates (i.e., detecting a signal when none was present) by recording mfVEPs with some of the sectors of the display occluded. In particular, the outer three rings (36 sectors) of the display were occluded so that only noise was recorded; false-positive rates for different values of SNR were calculated. The 2rSNR had the highest false-positive rate largely due to alpha in the records of some subjects. The mnSNR had a lower false-positive rate than did the inSNR because there was little correlation between the RMS of the noise in the signal-free window and the RMS of the noise within the signal window. Use of the mnSNR is recommended over the 2rSNR, especially where alpha contamination can not be eliminated. Ways to improve the SNR of the records are discussed. PMID- 12076019 TI - Formulation of a soil-pesticide transport model based on a compartmental approach. AB - A semianalytical soil-pesticide transport model is formulated based on a compartmental approach to determine spatial and temporal variations of pesticide residues across a soil profile. The compartmental model is implemented by drawing an analogy between a series of continuous-flow stirred tank reactors and a soil horizon that consists of multiple perfectly mixed compartments. The analogy is strengthened by exploiting a relation between the compartment series and the conventional convective-dispersive equation (CDE) for vertical transport in the soil. Consequently, the number of compartments in the model formulation is not free, but dictated as a function of transport parameters. The model formulation allows consideration of arbitrary boundary value specifications and also, for some cases, spatially varying initial concentration profiles. Sorption kinetics is represented via a two-site model that involves a linear sorption isotherm and a first-order irreversible sorption or a radial diffusive penetrating model. For these three cases, analysis of the compartmental model allows the resultant concentration profiles to be expressed in terms of the Poisson distribution. When a nonlinear kinetic sorption model is used to simulate the sorption processes, an analytical solution is not found and a numerical approach is required. PMID- 12076020 TI - Particle transport in unsaturated fractured chalk under arid conditions. AB - A series of field and laboratory experiments were conducted to study the mechanisms of particle detachment and transport from fractures in vadose chalk. Experiments of intermittent flow events along fracture surfaces were carried out in the laboratory. In the field, water was percolated from land surface via a discrete fracture into a compartmental sampler installed inside a horizontal corehole located I m below the surface. The mass, size distribution, and composition of the particles drained from the fracture voids were examined along with flow rates and salt dissolution. Two boreholes penetrating the underlying saturated zone were sampled and analyzed for colloidal concentration and composition. Most of the particle and solute release at the drained effluents occurred during the first several hours of flow, but erratic pulses of particles were still observed after long periods of time. Most of the detached particles had a mean diameter of >2 microm, while the mobile colloidal phase in the groundwater had a mean diameter of approximately 1 microm. Mineralogical composition of the groundwater colloids and the particles detached from the upper vadose fracture were similar. Laboratory observations demonstrated the importance of the existence of a coating layer, made of weathered particles and salts, on particle detachment. The results of this study suggest that: (1) particle detachment causes flow-rate variability in the unsaturated fracture; (2) the mechanisms of particle detachment and salt dissolution within the fracture are linked: and (3) although most of the detached particles are large and likely to accumulate inside fractures, some colloidal particles also eroded from the fracture void and are likely to be transported to the groundwater. PMID- 12076021 TI - Mobilization of small DNAPL pools formed by capillary entrapment. AB - We performed an experimental study to quantify the critical conditions for the mobilization of small pools of dense nonaqueous phase liquids (DNAPLs) that may form at capillary-heterogeneity boundaries. A series of experiments were conducted in columns packed with uniform sands arranged to create capillary heterogeneities. DNAPL pools readily formed in these packings and were more easily mobilized than trapped DNAPL ganglia. A model was developed to describe the critical conditions for DNAPL pool mobilization. Pool mobilization was expected when a dimensionless pool trapping number N(T)p> 1 while mobilization was observed in our experiments when N(T)p>0.76+/-0.16 (+/- 95% confidence interval). The difference between the model prediction and the experimental observations was attributed to experimental error. Using this model for DNAPL pool mobilization, a simple numerical experiment was conducted to illustrate use of the model and to explore the effect of scale on the critical conditions for pool mobilization. With an increase in system scale flow bypassing around the DNAPL pool increased and the system-averaged conditions for the onset of mobilization changed: a greater system-average Darcy flux or lower interfacial tensions were required for DNAPL pool mobilization. This result illustrates the importance of system scale on mobilization of DNAPL pools in systems with capillary heterogeneities. PMID- 12076022 TI - Computation of the interfacial area for two-fluid porous medium systems. AB - We develop a method to compute interfacial areas from three-dimensional digital representations of multiphase systems. We approximate the interfaces with the isosurface generated by the standard marching-cube algorithm from the discrete phase distribution. We apply this approach to two-fluid pore-scale simulations by (1) simulating a random packing of spheres that obeys the grain-size distribution and porosity of an experimental porous medium system, and (2) using a previously developed pore-morphology-based model in order to predict the phase distribution for a water-wet porous medium that undergoes primary drainage. The predicted primary drainage curve and interfacial areas are in good agreement with the experimental values reported in the literature, where interfacial areas were measured using interfacial tracers. The energy dissipation during Haines jumps is significant: thus, the mechanical work done on the system is not completely converted into surface energy, and interfacial areas may not be deduced from the primary drainage curve. PMID- 12076023 TI - Laboratory-scale in situ chemical oxidation of a perchloroethylene pool using permanganate. AB - In situ chemical oxidation (ISCO) is an emerging technology for the destruction of some chlorinated solvents present in subsurface environments. A laboratory investigation using a physical model was designed to assess the effectiveness of using permanganate as an oxidant to reduce the mass of a perchloroethylene (PCE) pool. The physical model was filled with silica sand overlying a silica flour base, simulating a two-dimensional saturated sand zone overlying a capillary barrier. PCE was introduced into the model so that it rested on top of the silica flour base, forming a dense nonaqueous phase liquid pool. The experimental methodology involved flushing the model with a permanganate solution for 146 days. During this period, measurements of chloride were used to assess the extent of pool oxidation. Before and after the oxidant flush, the quasi-steady state dissolution from the PCE pool was evaluated. Additionally, tracer studies were completed to assess changes in the flow field due to the oxidation process. At the termination of the experiment nine soil cores extracted from the model were used to detect the presence of MnO2 deposits and to quantify the mass of PCE remaining in the system. Excavation of the remaining material in the model revealed that the MnO2 distribution throughout the model was consistent with that observed in the cores. The oxidant flush was concluded before all of the pure phase PCE had been completely oxidized; however, approximately 45% of the PCE mass was removed, resulting in a fourfold decrease in the quasi-steady state aqueous phase mass loading of PCE from the pool. Measurements of chloride during the oxidant flush and of PCE in the soil cores suggested that the oxidation reaction occurred primarily at the upgradient edge of the PCE pool. MnO2 deposits within the model aquifer decreased the velocity of water directly above the pool, and the overall mass transfer from the remaining PCE pool. The results of this experimental study indicate that ISCO using permanganate is capable of removing substantial mass from a DNAPL pool; however, the performance of ISCO as a pool removal technology will be limited by the formation and precipitation of hydrous MnO2 that occurs during the oxidation process. PMID- 12076024 TI - Gel barrier formation in unsaturated porous media. AB - The gel barrier formation by a gelling liquid (Colloidal Silica) injection in an unsaturated porous medium is investigated by developing a mathematical model and conducting numerical simulations. Gelation process is initiated by adding electrolytes such as NaCl, and the gel phase consisting of cross-linked colloidal silica particles grows as the gelation process proceeds. The mathematical model describing the transport and gelation of Colloidal Silica (CS) is based on coupled mass balance equations for the gel mixture (the sol phase plus the gel phase), gel phase (cross-linked colloidal silica particles plus water captured between cross-linked particles), and colloidal silica particles (discrete and cross-linked) and NaCl in the sol (suspension of discrete colloidal silica particles in water) and gel phases. The solutions in terms of volumetric fraction of the gel phase yield the gel mixture viscosity via the dependency on the volumetric fraction of gel phase. This dependency is determined from a kinetic gelation model with time-normalized viscosity curves. The proposed model is verified by comparing experimentally and numerically determined hydraulic conductivities of gel-treated soil columns at different CS injection volumes. The numerical experiments indicate that an impermeable gel layer is formed within the time period twice the gel-point in a one-dimensional flow system. At the same normalized time corresponding to twice the gel-point, the CS solutions with lower NaCl concentrations result in further migration and poor performance in plugging the pore space. The viscosity computation proposed in this study is compared with another method available in the literature. It is observed that the other method estimates the viscosity at the mixing zone higher than the one proposed by the authors. The proposed model can simulate realistic injection scenarios with various combinations of operating parameters such as NaCl concentration and NaCl mixing time, and thus providing guidelines in performing this technology on site. PMID- 12076025 TI - Removal of As, Mn, Mo, Se, U, V and Zn from groundwater by zero-valent iron in a passive treatment cell: reaction progress modeling. AB - Three treatment cells were operated at a site near Durango, CO. One treatment cell operated for more than 3 years. The treatment cells were used for passive removal of contamination from groundwater at a uranium mill tailings repository site. Zero-valent iron [Fe(0)] that had been powdered, bound with aluminosilicate and molded into plates was used as a reactive material in one treatment cell. The others used granular Fe(0) and steel wool. The treatment cells significantly reduced concentrations of As, Mn, Mo, Se, U, V and Zn in groundwater that flowed through it. Zero-valent iron [Fe(0)], magnetite (Fe3O4), calcite (CaCO3), goethite (FeOOH) and mixtures of contaminant-bearing phases were identified in the solid fraction of one treatment cell. A reaction progress approach was used to model chemical evolution of water chemistry as it reacted with the Fe(0). Precipitation of calcite, ferrous hydroxide [Fe(OH)2] and ferrous sulfide (FeS) were used to simulate observed changes in major-ion aqueous chemistry. The amount of reaction progress differed for each treatment cell. Changes in contaminant concentrations were consistent with precipitation of reduced oxides (UO2, V2O3), sulfides (As2S3, ZnS), iron minerals (FeSe2, FeMoO4) and carbonate (MnCO3). Formation of a free gas phase and precipitation of minerals contributed to loss of hydraulic conductivity in one treatment cell. PMID- 12076026 TI - Biosynthesis of exopolysaccharide by a Bacillus licheniformis strain isolated from ropy cider. AB - A strain of Bacillus licheniformis displaying a ropy phenotype was isolated from a French ropy cider. The influence of culture conditions on the production of exopolysaccharide (EPS) was investigated. When B. licheniformis was grown in Man, Rogosa and Sharpe (MRS) medium, the highest amount of EPS was observed at mid exponential growth phase whatever the carbon source, glucose, fructose or sucrose. Interestingly at mid exponential growth phase, EPS amounts did not increase with increasing sugar concentrations. Incubation of B. licheniformis cells in media supplemented with ethanol (1-7%, v/v) revealed that EPS production was enhanced by the presence of ethanol, in exponential as well as in stationary phase. High Performance Liquid Chromatography (HPLC) and Nuclear Magnetic Resonance (NMR) analysis of EPS composition indicated that it was a heteropolymer in which mannose was the predominant monosaccharide as it constituted more than 80% of total polysaccharide. PMID- 12076027 TI - Gastrointestinal transit survival of an Enterococcus faecium probiotic strain administered with or without vancomycin. AB - The primary aim of this study was to evaluate if an ingested probiotic, containing viable Enterococcus faecium could survive gastrointestinal transit and if so, correlate the amount of the recovered probiotic strain with the host's own enterococci. The second aim was to investigate if simultaneous vancomycin intake influenced the survival and persistence of the probiotic strain and the stability of endogenous enterococci strains. Twenty healthy volunteers were given the probiotic product once daily for 10 days. Half of the subjects were simultaneously given vancomycin. Isolates of E. faecium strains were genotypically or phenotypically analysed with pulsed-field gel electrophoresis (PFGE) and the PhenePlate system, respectively. In eight of the ten volunteers given only the probiotic, the ingested E. faecium could be detected on day 10, while in none on day 31. From subjects given both probiotic and vancomycin no ingested E. faecium could be detected on day 10 or day 31. The estimated amount of ingested E. faecium recovered from faeces on day 10 ranged from 1.2 x 10(3) to 4.2 x 10(6) colony forming units per gram faeces, which in several cases were a substantial part of the total amount of E. faecium. The E. faecium isolated before probiotic plus vancomycin administration showed no close relationship to the ones isolated 3 weeks after ceased intake in any subjects. In conclusion, the ingested E. faecium strain can survive gastrointestinal transit. After intake, the E. faecium probiotic strain might become a large part of the total E. faecium population. The occurrence of the probiotic strain in the human gut seems to be transient after intake stop. Re-colonization of E. faecium after simultaneous probiotic plus vancomycin intake occurs mainly with strains without close genetic relationship to the strains harboured before treatment or to the ingested E. faecium strain. PMID- 12076028 TI - Inactivation and removal of Bacillus cereus by sanitizer and detergent. AB - This study was conducted to compare the susceptibility of planktonic cells, attached single cells and biofilm cells of Bacillus cereus to sodium hypochlorite and Spartec, a quaternary ammonium compound (QAC). Removal of B. cereus biofilm by simulating clean-in-place (CIP) procedures employing 1.5% Spec-Tak 1000, an alkaline detergent; 0.65% Dilac, an acid detergent and 1.5% Diverform Plus, a sodium hypochlorite containing alkaline detergent was also examined. The results show that B. cereus cells in a biofilm were most resistant to chemical sanitizers followed by the attached single cell and cells in a planktonic state. When B. cereus cells were in a planktonic state, 25 ppm hypochlorite or 100 ppm QAC induced a more than 5.0 log CFU/ml reduction of cell numbers within 15 s. However, the same sanitizer used in this study showed little effect on cells in a biofilm even when the exposure time was extended to 5 min. The sanitizers tested were less effective against cells in biofilm formed on milk pre-soiled than on unsoiled stainless steel chips. It was also noted that Spec-Tak 1000 treatment at 70 degrees C for 10 min and water rinse in the first step of the long-hot CIP procedure can effectively remove the biofilm B. cereus cells from the milk pre soiled stainless chip. With this long-hot CIP procedure, the biofilm cells of B. cereus were removed by ca. 6.10 log CFU/chip compared to ca. 1.47 log CFU/chip noted in the control procedure. PMID- 12076029 TI - The locus responsible for production of plantaricin S, a class IIb bacteriocin produced by Lactobacillus plantarum LPCO10, is widely distributed among wild-type Lact. plantarum strains isolated from olive fermentations. AB - The genes plsA and plsB encoding for production of plantaricin S (Pls), a two peptide bacteriocin produced by Lactobacillus plantarum LPCO10, are commonly distributed among wild-type Lact. plantarum strains isolated from olive fermentations. Among 68 independent isolates from different olive processing plants in South Spain, 15 of them were shown to produce bacteriocins that were active against other lactic acid bacteria, as well as spoilage and pathogenic bacteria. On the basis of PCR amplification and hybridization with specific probes, the Pls operon was detected in all the bacteriocin producer strains but not in the non-producer ones. Purification and subsequent amino acid sequencing of the bacteriocin produced by some of the 15 isolates yielded both the alpha and beta peptides from Pls. These results suggest that bacteriocin production contributes an ecological advantage for the wild-type Lact. plantanum strains in the colonization of the spontaneous, traditional olive fermentation process. PMID- 12076030 TI - Depuration dynamics of viruses in shellfish. AB - The consumption of shellfish has been associated with viral infections even in cases where shellfish complied with the current regulation, which is based on bacterial analysis. In this study, depuration rates of potential indicators and human viruses have been analysed in order to study the use of complementary parameters for evaluating the microbiological quality of depurated shellfish. Depuration of naturally highly polluted mussels has been evaluated and analyses for Escherichia coli, Clostridium perfringens, somatic coliphages, F-RNA phages and bacteriophages infecting Bacteroides fragilis RYC2056 and HSP40, human adenovirus, enterovirus have been done. Seawater of the depuration tank was disinfected by UV irradiation, ozone and passed through a skimmer and a biological filter. The correct functioning of the depuration tank was monitored by the quantification of total organic carbon (TOC), NH4+ and total aerobic bacteria in the seawater. To study the relation between the bacteriophages and the human viruses analysed, a logistic regression model was applied. F-RNA phages are significantly related to human adenoviruses and enteroviruses. Thus, they can be used as a complementary parameter for evaluating the efficiency of the depuration treatment. Somatic coliphages are also significantly associated with enteroviruses. Bacteriophages infecting B. fragilis HSP40 were analysed by the double-agar-layer (DAL) method, which quantifies infectious viruses, and by nested PCR, which detects the presence of the genome of these phages. The highest sensitivity of the molecular techniques was demonstrated and the results obtained are an indicator of a close relation between positive results by PCR and the presence of infectious viral particles in shellfish. All shellfish samples were negative for human viruses by PCR after 5 days of depuration treatment and the results obtained applying a regression model also showed negative results or nearly for F-RNA phages and bacteriophages infecting B. fragilis RYC2056. Thus, in this specific depuration treatment, 5 days may be necessary to assess the sanitary quality of shellfish. PMID- 12076031 TI - Pasteurization of milk and the heat resistance of Mycobacterium avium subsp. paratuberculosis: a critical review of the data. AB - Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) causes Johne's disease in ruminants (including cattle, sheep and goats) and other animals, and may contribute to Crohn's disease in humans. This possibility, and the fact that M. paratuberculosis may be present in raw milk, make it important to ensure that the heat treatment specified for pasteurization of milk will give acceptable inactivation of this bacterium, with an adequate margin of safety. Published studies of the heat resistance of this bacterium in milk have given widely differing results. Possible reasons for these differences, and the technical problems involved in the work, are reviewed. It is concluded that there is a need (i) for the adoption of an agreed Performance Criterion for pasteurization of milk in relation to this bacterium, (ii) a need for definitive laboratory experiments to understand and determine the heat resistance of M. paratuberculosis, and (iii) a need for an assessment of whether the minimum heat treatments specified at present for pasteurization of milk (Process Criteria) will meet the Performance Criterion for M. paratuberculosis. Measures are also required to ensure that commercial processes deliver continually the specified heat treatment, and to ensure that post-pasteurization contamination is avoided. PMID- 12076033 TI - Effect of freezing on the spontaneous fermentation and sensory attributes of Almagro eggplants. AB - A study of the physico-chemical and microbiological processes taking place during the spontaneous fermentation of eggplant fruits that had been frozen after blanching revealed a delay in the onset of the active fermentation stage, which has been ascribed to the adverse effect of the freezing temperature employed on the lactic acid bacteria involved in fermentation. The species most affected by freezing were L. fermentum and L. brevis biotype 2. Despite this finding, the pattern of fermentation was similar to the fermentation process in unfrozen eggplants. Freezing of the fruits did not significantly affect the sensory acceptability of the product and, in fact, in some cases, Almagro eggplants made from frozen fruits were preferred to commercially manufactured products. PMID- 12076032 TI - Empirical model building based on Weibull distribution to describe the joint effect of pH and temperature on the thermal resistance of Bacillus cereus in vegetable substrate. AB - A mathematical model based on Weibull parameters was built to describe the joint effect of temperature and pH on thermal inactivation of Bacillus cereus spores (strain INRA TZ415). The effect of these factors on Weibull model parameters (beta, 1/alpha) was also studied. Heat inactivation tests were carried out in acidified carrot broth as vegetable substrate, following a full factorial design at four levels for temperature (80, 85, 90 and 95 degrees C) and pH (6.2, 5.8, 5.2 and 4.7). The Weibull distribution model provided good individual fits for the different combinations of temperature-pH tested, with discrepancy factors, Df, coming close to 25% for most cases. The temperature and pH did not have a significant effect on the shape parameter (beta), which yielded a mean value of 0.88. The scale parameter (alpha) decreased with pH, and its inverse (1/alpha) followed an Arrhenius-type relationship with temperature. A global model was built, including the dependence of the alpha parameter on temperature and pH, and the model parameters were estimated by using a one-step nonlinear least-squares regression to improve the precision of the estimates. Results indicated that the global model provides a satisfactory description of the thermal inactivation of B. cereus spores, with R2 equal to 0.983. PMID- 12076034 TI - Numbers and types of microorganisms in vacuum-packed cold-smoked freshwater fish at the retail level. AB - Fifty-four packages (each one belonging to a different lot) of vacuum-packed cold smoked salmon (30) and trout (24) produced by six Spanish smokehouses were obtained at retail level after 3 weeks storage at 2+/-1 degrees C. Sensorial, chemical, physicochemical and microbiological characteristics were examined. Overall, pH, a(w), salt content in water phase, aerobic plate counts at 30 and 25 degrees C. levels of Enterobacteriaceae, lactic acid bacteria (LAB), fungi and presumptive aeromonads and staphylococci are in agreement with available data on lightly preserved fish products. Psychrotrophic clostridia ranged between 1.71 and 2.21 log CFU/g. Levels of ethanol were highly variable and not significantly related (p > 0.05) to sensory scores or to microbial numbers. Salmonella, Escherichia coli and Listeria monocytogenes were not detected in any sample. Listeriae other than L. monocytogenes were isolated from three packages. Levels of Staphylococcus aureus lower than 4 log CFU/g were also found in three packages. Among 377 bacteria randomly isolated from aerobic 25 degrees C plate counts, LAB predominated, with Carnobacterium (C. piscicola) and Lactobacillus (eight species) being the genera most frequently found. The second and third major groups were Enterobacteriaceae and Micrococcaceae, respectively. Proteus vulgaris, P. mirabilis and Serratia liquefaciens were dominant among Enterobacteriaceae and coagulase-negative staphylococci among Micrococcaceae. Minor microbial groups such as aerobic gram-negative bacilli (Acinetobacter; Moraxella and Pseudomonas), Brochothrix, Aeromonas, Bacillus and Vibrio constituted less than 17% of the total flora. PMID- 12076035 TI - Safety and efficacy of probiotic lactobacilli in promoting growth in post-weaning Swiss mice. AB - Lactobacillus rhamnosus MA27/6B and L. acidophilus MA27/6R are strains used in feed as probiotics. Their safety profiles and growth-stimulating properties were investigated via in vivo studies on young Swiss mice. After repeated administrations of different probiotic preparations in drinking water, safety parameters determined from liver, spleen and total weight remained unchanged. The growth-stimulating properties of viable or dead lactobacilli were studied after supplementation in drinking water. The feed intake (FI), water intake and body weight gain (WG) of the animals were compared to those of control mice. The lactobacilli supplementation of a sub-optimal diet made of barley allow recording of measurable growth performance of mice. It significantly increased WG compared to control groups (P < 0.01), by +28.9% and +31.7% for L. rhamnosus MA27/6B and L. acidophilus MA27/6R, respectively. This WG was correlated with a decrease in the consumption index. The effect of the dose ingested was also investigated: 10(8) lactobacilli CFU/mouse/day produced greater WG than 10(2), 10(4) or 10(6) CFU/mouse/day. No significant differences in growth performance parameters were observed between mice fed with 10(8) cells of viable or nonviable preparations. The mouse assay described could be used as a preliminary criterion when screening candidate probiotics for growth performance properties. PMID- 12076036 TI - A novel immunostimulating aspect of Lactobacillus gasseri: induction of "Gasserokine" as chemoattractants for macrophages. AB - The chemotactic activity of the culture supernatants from 14 strains of Lactobacillus acidophilus and L. gasseri was examined for murine macrophages. Significant macrophage chemotactic activity was observed in three strains of L. acidophilus and all strains of L. gasseri. The highest activity was observed in the supernatant (1131-sup) from 24-h cultures of L. gasseri JCM1131T. The chemotactic factor from 1131-sup, designated as "Gasserokine", was purified by the C18 reverse phase and ion-exchange chromatography. The purity of Gasserokine was checked by HPLC with the reverse-phase mode. The chemotactic activity of Gasserokine was also observed for human monocytes. The macrophage chemotaxis induced by L. gasseri JCM1131T culture supernatants was discovered to be a new biological function exerted by probiotic lactic acid bacteria. Therefore, the activity is expected to be used for one of the functional parameters in the immunomodulating properties of probiotic lactic acid bacteria. PMID- 12076037 TI - A simple, spreadsheet-based, food safety risk assessment tool. AB - The development and use of a simple tool for food safety risk assessment is described. The tool is in spreadsheet software format and embodies established principles of food safety risk assessment, i.e., the combination of probability of exposure to a food-borne hazard, the magnitude of hazard in a food when present, and the probability and severity of outcomes that might arise from that level and frequency of exposure. The tool requires the user to select from qualitative statements and/or to provide quantitative data concerning factors that that will affect the food safety risk to a specific population, arising from a specific food product and specific hazard, during the steps from harvest to consumption. The spreadsheet converts the qualitative inputs into numerical values and combines them with the quantitative inputs in a series of mathematical and logical steps using standard spreadsheet functions. Those calculations are used to generate indices of the public health risk. Shortcomings of the approach are discussed, including the simplifications and assumptions inherent in the mathematical model, the inadequacy of data currently available, and the lack of consideration of variability and uncertainty in the inputs and outputs of the model. Possible improvements are suggested. The model underpinning the tool is a simplification of the harvest to consumption pathway, but the tool offers a quick and simple means of comparing food-borne risks from diverse products, and has utility for ranking and prioritising risks from diverse sources. It can be used to screen food-borne risks and identify those requiring more rigorous assessment. It also serves as an aid to structured problem solving and can help to focus attention on those factors in food production, processing, distribution and meal preparation that most affect food safety risk, and that may be the most appropriate targets for risk management strategies. PMID- 12076038 TI - A semi-quantitative seafood safety risk assessment. AB - As part of a semi-quantitative risk assessment of 10 seafood hazard/product combinations, a risk assessment tool was used to generate a Risk Ranking. The tool is in a spreadsheet software format and provides a risk estimate, which is scaled between 0 and 100, where 0 represents no risk and 100 represents all meals containing a lethal dose of the hazard. A full description of the tool is contained in Ross and Sumner (this issue). Based on their ranking, seafoods in Australia fell into three risk categories. Hazard/product pairs with ranking < 32 included mercury poisoning (Relative Risk = 24), Clostridium botulinum in canned fish (RR = 25), or in vacuum-packed cold-smoked fish (RR = 28), parasites in sushi/sashimi (RR = 31), viruses in shellfish from uncontaminated waters, (RR = 31), enteric bacteria in imported cooked shrimp (RR = 31) and algal biotoxins from controlled waters (RR = 31). It is noted that there have been no documented cases of food-borne illness from any of the above hazard/product pairings in Australia. Those with rankings 32-48 included Vibrio parahaemolyticus in cooked prawns (RR = 37), V. cholerae in cooked prawns (RR = 37), Listeria monocytogenes in cold-smoked seafoods (RR = 39), scombrotoxicosis (RR = 40), V. vulnificus in oysters (RR = 41), ciguatera in the general Australian population (RR = 45), L. monocytogenes in susceptible (RR = 45) and extremely susceptible populations (RR = 47) and enteric bacteria in imported cooked shrimp eaten by vulnerable consumers (RR = 48). Almost all the hazard/product pairs in this category have caused the outbreaks of food poisoning in Australasia. Those hazard/product pairs with rankings >48 included ciguatera from recreational fishing in susceptible areas (RR = 60), viruses in shellfish from contaminated waters (RR = 67) and algal biotoxins from uncontrolled waters in an algal event (RR = 72). There have been significant (>100 cases) food poisoning incidents involving viruses and biotoxins in shellfish, while ciguatera poisoning is prevalent among coastal communities in Australia's warmer waters. PMID- 12076039 TI - Staphylococcal enterotoxin genes of classical and new types detected by PCR in France. AB - The PCR detection of the genes coding for the newly described staphylococcal enterotoxins (SE) G, H, I and J was carried out for 332 foodborne staphylococci, isolated from a variety of foods in France. The frequency of the Staphylococcus aureus strains harboring these genes was found to be very high (57%) and greater than that of the strains harboring "classical" SE genes as previously established. If one takes into account the newly described SE genes, in addition to the classical SE genes, the percentage of foodborne enterotoxigenic S. aureus strains doubles. The S. aureus biovars that were rarely or never enterotoxigenic (i.e., the poultry and bovine biovars) frequently become more potentially toxigenic, if taking into account the seg. seh, sei and sej genes. Furthermore, this work indicated a systematic association between the seg and sei genes and their high incidence among the S. aureus strains, which suggests that these two SE's could be an important phylogenetic link among the staphylococcal enterotoxins. PMID- 12076040 TI - Alicyclobacillus acidoterrestris spores as a target for Cupuacu (Theobroma grandiflorum) nectar thermal processing: kinetic parameters and experimental methods. AB - The kinetic parameters of thermal inactivation of a spore former, Alicyclobacillus acidoterrestris, in a tropical fruit nectar [25% of Cupuacu (Theobroma grandiflorum) pulp and 15% sugar] were determined by the isothermal method (IM), under batch heating, and by the paired equivalent isothermal exposures (PEIE) method, under non-isothermal continuous conditions. The isothermal experiments were repeated three times, every 4 months, with the same spore suspension kept frozen between experiments. The aging of spores, under frozen storage, seemed to produce a notorious increase in the z-value from experiment to experiment: Experiment 1 (z = 7.8 +/- 2.6 degrees C, D(95 degrees C) = 5.29 +/- 0.96 min), Experiment 2 (z = 22 +/- 5 degrees C, D(95 degrees C) = 5.99 +/- 0.63 min), and Experiment 3 (z =29 +/- 10 degrees C, D(95 degrees C) = 3.82 +/- 0.48 min). The evaluation of the kinetic parameters by the PEIE method was carried out in parallel with Experiment 3, with the same aged spores, and the results (z = 31 +/- 6 degrees C, D(95 degrees C) = 5.5 +/- 1.2 min) were close to the ones obtained in this experiment. From this work, it seems that the PEIE method can also be applied to evaluate the reduction parameters of a spore forming microorganism, and in a more realistic way, since the continuous system eliminates the errors caused by come-up and cool-down times (CUT and CDT) that are unavoidable in isothermal experiments. Therefore, when designing a thermal process for a continuous system, the PEIE method should be used, or the chances are that the process would be underdesigned, risking that the desired level of spore inactivation would not be achieved. An optimization of the thermal processing conditions was next performed for Cupuacu nectar, considering a 5D reduction in A. acidoterrestris spores. If a pasteurization process is considered, the conditions that ensure safety (9 min at 98 degrees C) only allow a 55% retention of ascorbic acid (AA). If sterilization is considered, 8 s at 115 degrees C will ensure a safe product and retain 98.5% of the original ascorbic acid. Therefore, if A. acidoterrestris is considered as the target microorganism, the nectar should undergo an aseptic high temperature short time principle (HTST) process to achieve a 5D reduction in this acidophilus spore former. However, if the hot-fill-and-hold pasteurization process is preferred, the product should be fortified with ascorbic acid. PMID- 12076041 TI - Similar Listeria monocytogenes pulsotypes detected in several foods originating from different sources. AB - The purpose of the study was to obtain fingerprinting data of Listeria monocytogenes strains isolated in various foods to determine possible associations of strains with product type, producer, country or isolation time. Two hundred and ninety-five L. monocytogenes strains originating from food items of 41 producers of 10 countries were characterized by pulsed-field gel electrophoresis (PFGE) typing. Combination of AscI and ApaI macrorestriction patterns (MRP) yielded 66 different pulsotypes. Ten pulsotypes were common to two or more product types and 17 pulsotypes were detected in foods of more than one producer having no apparent association with each other. Similar pulsotypes of L. monocytogenes were recovered in products of different countries over several years. Some of the pulsotypes were recurrently recovered from the same product of the same producer, suggesting a possible persistence of these strains in the processing plant. However, some of the recurrently isolated L. monocytogenes pulsotypes were repeatedly found in products of several producers, which may indicate that persistent houseflora strains are not always producer-specific. Furthermore, the similarity of macrorestriction patterns expressed as clusters, based on the numerical analysis of macrorestriction patterns, was not found to correlate with product type, country, producer or year of isolation. Our data suggest a wide geographical and temporal distribution of a number of L. monocytogenes strains isolated in food products. The existence of similar L. monocytogenes strains in various food products of several producers should be considered if food strain fingerprint results are used to help trace the vehicles for infections. PMID- 12076042 TI - Effects of cutting process on pork meat contamination by verotoxin-producing Escherichia coli (VTEC) and E. coli O157:H7. AB - The aims of the present study were: (i) to evaluate verotoxin-producing Escherichia coli (VTEC) prevalence in pork cutting meat; (ii) to determine the effects of cutting process on pork meat contamination by VTEC; (iii) to characterise the VTEC strains isolated from pork and pork cutting plants (virulence genes and serotype); and (iv) to compare the strains isolated the same day in the same cutting plant in order to identify the routes of contamination inside the cutting plant. Pork carcasses from three French cutting plants were sampled before carcass cutting (carcass samples), after carcasses were divided into big portions (untrimmed cuts) and after preparation of primal cuts (rindless boneless cuts), and different environmental sites in each cutting plant were sampled at three different times in the work day. Potable water was also collected. PCR detection of stx genes was performed on a total of 2042 samples. In addition, a second PCR specific for E. coli O157:H7 detection was carried out on the stx-positive samples. VTEC strains were recovered from positive samples by colony hybridisation or immunoconcentration, then serotyped, genetically characterised (eae, ehx, stx1, stx2, stx2e, uidA and genes which are associated with virulence) and pulsotyped. No E. coli O157:H7 was detected. Meat contamination decreased from carcass (12%) and primary cuts (19%) to secondary cuts (5%), whereas environmental contamination increased after 2 h of activity (from 3% before the commencement of the work day to 25% and 20%, 2 and 6 h after commencement of cutting). No VTEC isolates harboured eae, ehx and uidA genes. VTEC contamination routes were not clearly identified. PMID- 12076044 TI - Genetic management of nonhuman primates. AB - Genetic management is widely recognized as a critical component of the overall management of captive nonhuman primate colonies which produce animals for biomedical research. In this paper, we review the roles of conservation-oriented genetic management, research-oriented genetic management, genetic management at the level of taxomomic class, genetic management at the level of the population, and quantitative genetic analysis in comprehensive genetic management programs for nonhuman primate colonies. We conclude that genetic management is crucial for maintaining nonhuman primate populations suitable for genetic research on normal and disease-related phenotypes. In addition, for research programs that do not have specific genetic objectives, genetic management is essential to facilitate the selection of samples of well-matched unrelated animals for experimental purposes. PMID- 12076043 TI - Effects of slaughter processes on pig carcass contamination by verotoxin producing Escherichia coli and E. coli O157:H7. AB - The aims of the present study were: (i) to evaluate verotoxin-producing Escherichia coli (VTEC) faecal carriage of slaughtered pigs; (ii) to determine the effects of three different pig slaughtering processes on pig carcass contamination by VTEC; (iii) to characterise the VTEC strains isolated from pig and pig slaughterhouses (virulence genes and serotype); and (iv) to compare the strains isolated in the same slaughterhouse in order to identify the routes of contamination inside the slaughterhouse. Pork carcasses from three French slaughterhouses were sampled at three steps of the slaughter process and different sites in each slaughterhouse were sampled at three different times in the work day. Faecal material from each sampled carcass, potable water and scalding water were also collected. Detection of stx genes was performed by polymerase chain reaction (PCR) on a total of 1227 samples. In addition, a second PCR specific for E. coli O157:H7 detection was carried out on the stx-positive samples. VTEC strains were recovered from positive samples by colony hybridisation or immunoconcentration, then serotyped, genetically characterised (eae, ehx, stx1, stx2, stx2c, uidA genes associated with virulence) and pulsotyped. No E. coli O157:H7 was isolated from the three uidA-positive samples. VTEC faecal carriage was 31%. Global carcass contamination decreased with slaughter process (from 46% to 15%), whereas environmental contamination increased (from 7% to 29%). No VTEC isolates harboured eae, ehx, and uidA genes. VTEC contamination routes were not clearly identified. PMID- 12076045 TI - The FIC1 gene: structure and polymorphisms in baboon. AB - A genome scan performed on 648 pedigreed baboons to detect and localize quantitative trait loci (QTL) for lipoprotein phenotypes that are known risk factors for atherosclerosis indicated the presence of a QTL on chromosome 18q that exerts a major influence on HDL-cholesterol (HDL-C) related phenotypes. Inspection of the human gene map revealed that the familial intrahepatic cholestatis gene 1 (FIC1) maps to the homologous region of baboon chromosome 18 containing the major QTL influencing HDL-C phenotypes. FIC1 is a strong biological candidate for this QTL because HDL-C is the preferred precursor for bile acid synthesis. In this study, we cloned and sequenced FIC1 cDNA and found that it is highly conserved between human and baboon. We also sequenced FIC1 cDNAs from a panel of unrelated baboons revealing single nucleotide polymorphisms (SNPs) and a polymorphic dinucleotide repeat. None of the baboon SNPs corresponded to human FIC1 mutations associated with familial intrahepatic cholestasis or benign recurrent intrahepatic cholestasis disorders. PMID- 12076046 TI - Viral dynamics of early HIV infection in neonatal macaques after oral exposure to HIV-2287: an animal model with implications for maternal-neonatal HIV transmission. AB - A model of vertical HIV transmission was developed using oral HIV-2(287) exposure of newborn Macaca nemestrina. The minimal Animal Infectious Dose for this oral route was found to be 10-fold higher than that for atraumatic viral transmission across other mucosal membranes (vaginal/rectal) of juvenile macaques. However, once infection was established, viral replication was rapid and plasma viremia could be detected by reverse-transcriptase polymerase chain reaction and viral co culture within 1 week following exposure. No animal was resistant to infection and all macaques initially exposed to a subinfectious viral inoculum were subsequently infected by re-exposure of mucosal membranes. Higher viral load during primary infection correlated with a more rapid CD4 depletion; however, all HIV-2(287)-infected animals developed CD4 depletion during the observation period. This animal model can now be used to study early viral replication in the presence and absence of anti-retroviral agents to help identify conditions to reduce vertical HIV transmission in human newborns. PMID- 12076047 TI - DNA prime/protein boost vaccine strategy in neonatal macaques against simian human immunodeficiency virus. AB - Newborn macaques were vaccinated against a chimeric simian human immunodeficiency (SHIV) virus, SHIV-vpu+, by DNA priming and boosting with homologous HIV-1 gp160. Following SHIV-vpu+ challenge, containment of infection was observed in 4 of 15 animals given DNA priming/protein boost vaccination and in three of four animals given gp160 boosts only. Rechallenge with homologous virus of six animals that contained the first challenge virus resulted in rapid viral clearance or low viral loads. Upon additional rechallenge with heterologous, pathogenic SHIV89.6P, four of these six animals maintained normal CD4+ T-cell counts with no or limited SHIV89.6P infection. Our data suggest that humoral and cellular immune mechanisms may have contributed to the containment of SHIV89.6P; however, viral interference with SHIV-vpu+ could also have played a role. Our results indicate that immunogenicity and efficacy of candidate AIDS vaccines are not affected when vaccination is initiated during infancy as compared with later in life. PMID- 12076048 TI - Polymorphism in the rhesus macaque (Macaca mulatta) NRAMP1 gene: lack of an allelic association to tuberculosis susceptibility. AB - Although previous tuberculosis (TB) research has suggested that underlying genetic factors influence a host's response and ability to survive Mycobacterium infection, only recently has a gene been identified, the 'natural resistance associated macrophage protein 1' (NRAMP1) gene, which provides a degree of natural resistance to infection by some Mycobacterium species. To date, however, the role that NRAMP1 may play in resistance to Mycobacterium infection has only been examined in mouse and man. Here, we present data generated at NRAMP1 among a group of rhesus macaques (Macaca mulatta) that were euthanized because of an outbreak of Mycobacterium tuberculosis during quarantine. Data were also generated on unrelated (and healthy) rhesus macaques in order to better determine the frequency and degree of genetic polymorphism within Macaca at the NRAMP1 locus. These data represent the first study designed to examine the role that NRAMP1 may play in TB susceptibility among rhesus macaques. PMID- 12076049 TI - Survey of dental radiographic services in private dental clinics in Damascus, Syria. AB - OBJECTIVES: To perform a radiographic survey of private dental clinics in Damascus, Syria using a postal questionnaire to produce recommendations for improving the quality of dental radiographic services and education in Syria. METHODS: Three hundred private dental clinics in Damascus were surveyed using a postal questionnaire (in English and Arabic) containing 27 questions on demographic information, equipment, techniques, selection criteria, frequency of examinations, and undergraduate/ postgraduate education. RESULTS: Two hundred and two (67%) dentists responded of which 95% graduated in Syria. The results showed a general lack of knowledge and understanding of dental radiography. Sixty four per cent did not know the kVp setting of their equipment, 73% used D-speed film, 57% did not use film holders and beam aiming devices, 25% did not use a viewing box. In addition, 45% of known equipment operated at 50 kVp or less and 16% was over 20 years old. No meaningful selection criteria existed with a wide variation in type and frequency of radiographs used for different clinical conditions. Syrian undergraduate training in dental radiology was minimal and there was no postgraduate education in the speciality. CONCLUSIONS: Several areas of the radiographic service in Damascus fall short of current recommendations on good practice. Recommendations are made to improve the service, the quality of undergraduate education and to establish postgraduate education. PMID- 12076050 TI - Impact of JPEG lossy image compression on quantitative digital subtraction radiography. AB - OBJECTIVES: The aim of the study was to evaluate the impact of JPEG lossy image compression on the estimation of alveolar bone gain by quantitative digital subtraction radiography (DSR). METHODS: Nine dry domestic pig mandible posterior segments were radiographed three times ('Baseline', 'No change', and 'Gain') with standardized projection geometry. Bone gain was simulated by adding artificial bone chips (1, 4, and 15 mg). Images were either compressed before or after registration. No change areas in compressed and subtracted 'No change-Baseline' images and bone gain volumes in compressed and subtracted 'Gain-Baseline' images were calculated and compared to the corresponding measurements performed on original subtracted images. RESULTS: Measurements of no change areas ('No change Baseline') were only slightly affected by compressions down to JPEG 50 (J50) applied either before or after registration. Simulated gain of alveolar bone ('Gain-Baseline') was underestimated when compression before registration was performed. The underestimation was bigger when small bone chips of 1 mg were measured and when higher compression rates were used. Bone chips of 4 and 15 mg were only slightly underestimated when using J90, J70, and J50 compressions before registration. CONCLUSIONS: Lossy JPEG compression does not affect the measurements of no change areas by DSR. Images undergoing subtraction should be registered before compression and if so, J90 compression with a compression ratio of 1:7 can be used to detect and measure 4 mg and larger bone gain. PMID- 12076051 TI - Detection of approximal caries in digital radiographs before and after correction for attenuation and visual response. An in vitro study. AB - OBJECTIVE: To evaluate if digital compensation for exponential attenuation and the characteristics of the human visual system improves the diagnosis of approximal caries from digital radiographs. MATERIAL AND METHODS: Forty premolar teeth were mounted in plaster blocks. Radiographs of the teeth were exposed with the Dixi digital intraoral system employing a Prostyle Intra dental X-ray unit (Planmeca Oy, Helsinki, Finland). Thirteen radiographs were then processed to compensate for the exponential attenuation and for the characteristic of the human visual system using equations presented in the paper. Ten observers were asked to diagnose approximal caries in all radiographs. ROC analyses were performed. The teeth were subsequently sectioned for histological validation of the lesions. The areas under ROC curves of original and processed radiographs were compared and analysed using Wilcoxon's signed-ranks test. RESULTS: There were significant diagnostic differences between the two types of radiographs (all lesions P=0.005. enamel P=0.028, and dentine P=0.050). CONCLUSION: Digital radiographs processed to compensate for exponential attenuation and the characteristics of the human visual system significantly improves the diagnosis of approximal caries in vitro. PMID- 12076052 TI - Correction for attenuation and visual response in digital radiography. AB - OBJECTIVES: To derive a method to correct for the exponential attenuation adding visual linearization for digital radiography and to perform an experimental test to study effects on observer performance. METHOD: A theoretical analysis was performed and expressions were derived to correct radiographic data for the attenuation. An experiment was performed exposing an aluminum step wedge with holes of increasing depths behind each step plus soft tissue simulation. Seven original images were created with a digital intraoral X-ray sensor and a further 14 transformed images produced. Ten observers examined the images in a randomized order reporting the number of holes seen per wedge step. The results were analysed by One Way ANOVA. RESULTS: It was possible to correct for attenuation and the response of the human visual system to light intensities from a computer monitor. Perception was significantly improved in all recalculated radiographs that took both attenuation and the response of the eye into account (P-values ranged from <0.0001 to <0.0286). CONCLUSIONS: The transforms may be useful for improved perception when viewing digital radiographs. PMID- 12076054 TI - Visual linearization of the display of digital radiographs. AB - OBJECTIVES: To derive and test a method to linearize the visual response of the display of digital radiographs so that equal steps in gray levels will be perceived as equal steps in brightness. METHOD: A mathematical analysis was performed and expressions for visual linearization were derived. Twenty-four test images were computer generated to confirm that visual linearization may be achieved. Each image had three groups of square areas of different size placed in three rows. The left and right squares in each row were given different gray levels to simulate various contrast levels. The middle squares were initially given the same gray level value as one of the outer squares. The images were examined by ten observers who could change the gray levels of the middle squares so that the step in brightness between the middle square and the outer squares become subjectively equal. The test was performed three times employing two different monitors. RESULTS: The experimental test confirmed that visual linearization could be achieved. Linear regression analyses gave determination coefficients of 0.9926 amd 0.9942 for monitors with gamma-values of 1.93 and 2.50. respectively. The mean data from the ten observers perfectly fitted those theoretically calculated. CONCLUSION: Visual linearization of gray levels can be achieved but further clinical research is needed to determine if this improves diagnosis. PMID- 12076053 TI - Clinical indications for digital volume tomography in oral and maxillofacial surgery. AB - Digital volume tomography (DVT, NewTom, NewTom AG, Marburg, Germany) is a novel technique for maxillofacial imaging at a lower radiation dose and lower cost than CT. We describe four clinical cases to illustrate its potential advantages. PMID- 12076055 TI - Thin-plate spline (TPS) graphical analysis of the mandible on cephalometric radiographs. AB - We describe two cases of Class III malocclusion with and without orthodontic treatment. A thin-plate spline (TPS) analysis of lateral cephalometric radiographs was used to visualize transformations of the mandible. The actual sites of mandibular skeletal change are not detectable with conventional cephalometric analysis. These case analyses indicate that specific patterns of mandibular transformation are associated with Class III malocclusion with or without orthopaedic therapy, and visualization of these deformations is feasible using TPS graphical analysis. PMID- 12076056 TI - Bilateral coronoid hyperplasia in two brothers. AB - Coronoid hyperplasia is a rare condition of unknown aetiology that can occur in both unilateral and bilateral forms. Without radiographic investigation the diagnosis is often missed. Researchers have postulated a familial form of inheritance. This study reports the occurrence of coronoid hyperplasia in two brothers. The parents were unaffected and there are no other siblings. The diagnosis was confirmed with the aid of panoramic radiographs and axial computed tomographic scans with para-sagittal reconstructions which demonstrated enlargement of the coronoid processes and in one case impingement against the zygomatic bone. One brother was successfully treated with a unilateral intra-oral coronoidectomy whilst the other was unsuccessfully treated with a bilateral intra oral coronoidectomy. PMID- 12076057 TI - Inflammatory orbital pseudotumor with infratemporal fossa extension mimicking temporomandibular joint dysfunction. AB - Inflammatory orbital pseudotumor is a benign space-occupying lesion of unknown origin that involves all or part of the fatty tissue within the orbit. Occasionally the disease may extend into the middle cranial, the pterygopalatine, and the infratemporal fossa through the various foramina of the orbit, although extension into the infratemporal fossa is very rare. We present a case which extends into the infratemporal fossa through the inferior orbital fissure, resulting in presenting symptoms mimicking temporomandibular joint dysfunction. PMID- 12076058 TI - Malignant melanoma of mandibular gingiva; the usefulness of fat-saturated MRI. AB - Case report of a histologically validated malignant melanoma affecting the mandibular molar gingiva in a 31-year-old woman. Tumour was evaluated by magnetic resonance (MR) imaging at 1.5-T, spin-echo 3 mm slice thickness with T1-weighted and fat-saturated T1-weighted images. The fat-saturated T1-weighted images demonstrated the lesion more clearly than conventional T1-weighted images. PMID- 12076059 TI - Late allergic reaction following sialography. PMID- 12076060 TI - Odontogenic myxomas in the Hong Kong Chinese: clinico-radiological presentation and systematic review. AB - AIM: The aim of this study was to determine the clinical and radiological presentation on plain films of central odontogenic myxomas (OM) in the Hong Kong Chinese and compare them to other reported series by a systematic review (SR). METHODS: The study had two elements, that of a complete series of all OMs presenting at a major Chinese maxillofacial surgical unit and a SR. The files of the Department of Oral and Maxillofacial Surgery of the University of Hong Kong between 1989 and 2000 were reviewed for OM cases. The relevant literature was identified by electronic databases, review of citation lists and handsearching of key journals. The principal selection criterion was that the study should represent a complete collection of cases. RESULTS: The SR revealed that only a quarter of the reports provided any quantitative radiological details. Just under half of the included reports provided the relative period prevalence of OMs compared to other odontogenic tumours, but these reports almost completely excluded consideration not only of the radiology but also specific clinical details. Furthermore, they did not report of recurrence data, an important feature of the OM. Most of the most recent reports were concerned with relative period prevalence. The relative period prevalence and 'number of OMs per hospital per year' of studies separated by two or more decades in Hong Kong and Nigeria indicate an increase in the number of lesions diagnosed as OMs. The 10 Hong Kong cases were broadly consistent with the predilections for females and the mandible of other reports. The mean age at first presentation in the present report is 36.9, older than the other reports. Most lesions are large, affecting both posterior and anterior quadrants. They appear to be larger than those in many other reports. Two of the OMs are accompanied by histories of recent rapid growth. Tooth displacement and root resorption are more frequently observed in OMs in the present study. Although all OMs in the present study are still being followed up after surgery, none have recurred. CONCLUSION: Although the presentation of larger lesions in the older Chinese could in part be explained by attitudes rooted in traditional medicine in spite of the widespread availability of modern medical care in Hong Kong, there is a history of recent rapid growth in at least two cases. An emphasis by recent reports on the OMs' relative period prevalence of concurrent odontogenic lesions, has omitted recording the radiology and the specific clinical details usually required for diagnosis., PMID- 12076061 TI - Oral radiographic predictors of osteoporosis. AB - Osteoporosis is a systemic skeletal disease characterized by low bone mass and micro architectural deterioration of the bone scaffold that result in increased bone fragility and susceptibility to fracture. It is one of the most common disorders of the elderly and is estimated to effect 75 million people in Europe, Japan and the USA. The purpose of screening for osteoporosis is to identify individuals who are likely to benefit from treatment. The fact that dental radiographs are regularly made on a large fraction of the adult population makes their potential use as a marker of skeletal health an exciting avenue of research. In the last four decades numerous research teams have reported oral radiographic findings associated with osteoporosis. The preponderance of the evidence shows that the jaws of subjects with osteoporosis show reduced bone mass and altered morphology. Clinically useful predictions of individuals most likely to develop osteoporotic fractures will require a multifactorial model including both radiographic and clinical findings. Future efforts should continue to search for oral radiographic signs with high sensitivity and specificity for osteoporosis, identify clinical signs available in the dental office associated with osteoporosis, develop multidisciplinary classification methods including both radiographic and clinical parameters, and automate radiographic and clinical analyses as much as possible to minimize the involvement of the dentist as well as to standardize data collection. PMID- 12076062 TI - A comparison of digital and film radiography in Dutch dental practices assessed by questionnaire. AB - OBJECTIVES: The purpose of this study was to assess the experiences of general dental practitioners (GDPs) with solid-state and storage phosphor digital sensors compared to film. METHODS: In a mail survey 578 questionnaires were sent to Dutch GDPs who were users of digital X-ray detectors and film. The questionnaire requested demographic data, information about the digital system used and the user-friendliness of the X-ray detector system. In the analysis of the data these variables were related to the type of system used and also to the demographic data. The data were analysed using descriptive statistics, variance-analysis (One Way ANOVA) and nonparametric tests (Kruskal-Wallis, Mann-Whitney and Wilcoxon) (SPSS 9.0). RESULTS: Four hundred and twenty-four questionnaires were returned (overall response rate 73%). The user-friendliness of the handling of the different systems before exposing the radiograph is better for conventional film, whereas the handling after exposing the radiograph favoured the digital X-ray detector systems. CONCLUSIONS: The user-friendliness was best for film pre exposure and digital sensors post-exposure. PMID- 12076063 TI - Lessons from similarities between SLE and HIV infection. AB - OBJECTIVE: We attempted to obtain deeper understanding of systemic lupus erythematosus (SLE) and human immunodeficiency virus (HIV) infection through comparative studies between both diseases. METHOD: For this purpose, we reviewed and discussed lessons from similarities in both diseases based on our own and reported findings in literatures. RESULT: Such comparative studies may contribute to the progress in understanding the clinical or pathogenetic features of these diseases. CONCLUSION: Further studies into the relationship between SLE and HIV infection may bring to light important clues to assist in the development of better treatments for each disease. PMID- 12076064 TI - Latency and reactivation of human cytomegalovirus. AB - The sequence analysis of herpesviruses suggests they have been evolving with their individual vertebrate hosts for millions of years, and their divergence parallels that of the hosts they infect. Given this time they have been learning to live with their individual hosts, it is not surprising that they have become extremely well adapted to doing so without causing much in the way of obvious disease. A key feature of their strategy for persisting in the host is the ability of all herpesviruses to establish latent infection-a state in which no, or only a very limited set of, viral genes are expressed in cells in which viral DNA persists. The alpha herpesviruses (herpes simplex and varicella zoster virus) establish latency in neuronal cells in sensory ganglia: these are long lived non dividing cells and the alpha herpesviruses persist in these with expression of only the latency associated transcripts-although the function of these RNA transcripts remains incompletely understood. The principal gamma herpesvirus of humans, Epstein Barr virus (EBV), is latent mainly in B lymphocytes: EBV persistence in B cells is associated with expression of a limited set of viral genes encoding functions necessary for the maintenance of the episomal viral DNA as B cells divide.The mechanism by which the principal beta herpesvirus of humans human cytomegalovirus (HCMV) persists, is also incompletely understood and the subject of this review. Understanding how HCMV persists has clinical relevance in that its transmission to seronegative recipients might be more easily prevented, and the mechanisms by which it produces disease in the neonate and immunocompromised hosts more easily understood, if we knew more about the cells in which the virus is latent and the way in which it reactivates. PMID- 12076065 TI - Clinical aspects and management of cytomegalovirus infection. AB - The previous article discussed recent advances in understanding the biology of HCMV infection. Here we discuss current approaches to the clinical management of HCMV disease, and how understanding the biology of the virus may affect these. PMID- 12076066 TI - Epidemiology of pneumonia hospitalizations in Spain, 1995-1998. AB - OBJECTIVES: This epidemiological survey was undertaken to estimate the burden of hospital admissions for pneumonia in Spain during a four-year period (1995-1998). METHODS: Data were obtained from the national surveillance system for hospital data. RESULTS: There were 231,512 hospital admissions for pneumonia (ICD 9 CM 480 486; first listed diagnosis) during this period, that is an annual incidence of 177 cases per 100,000 population. The incidence was higher in children <5 years of age and in persons >or=65 years compared with other age groups. The annual cost of these hospitalizations to the National Health Care System was of 127 million ECUS. CONCLUSIONS: Preventive measures, such as vaccination of population groups at high risk, to reduce pneumonia-related morbidity could result in large cost savings to the National Health Care System. PMID- 12076068 TI - Treatment of severe infectious mononucleosis with famciclovir. AB - We report a patient with severe acute infectious mononucleosis who was successfully treated with famciclovir. A 15-year-old male was admitted with a 6 week history of fever, malaise, generalized lymphadenopathy, and hepatosplenomegaly, the patient was acutely ill with a temperature of 39.0 degrees C. Oropharingeal examination revealed enlarged tonsils partially obstructing the airways. EBV serology obtained during admission showed a positive Monospot test, virus capsid antigen IgM, 1:320, Epstein-Barr nuclear and early antigen, negative. After 72 hours of treatment with famciclovir (500 mg t.i.d.), the patient was afebrile with important regression of the lymphadenopathy, enlarged tonsils and hepatosplenomegaly. Because acute infectious mononucleosis may be associated with extensive and prolonged disease, the potential therapeutic role of famciclovir in the treatment of severe forms of the disease deserves further studies. PMID- 12076067 TI - Low prevalence of antiretroviral drug resistance among HIV-1 seroconverters in London, 1984-1991. AB - OBJECTIVES: To examine the prevalence of resistance mutations and natural polymorphisms to reverse transcriptase (RT) and protease inhibitors in a cohort of patients with defined seroconversion dates. METHODS: Eligible patients were those attending an HIV centre in North London who seroconverted from HIV negative to positive status between 01/01/85 and 31/12/91 (n=104). Genotypic resistance analysis was performed on the first positive serum sample after seroconversion and before use of antiretroviral therapy using population-based sequencing of RT PCR fragments and rule-based sequence interpretation (Vircogen). RESULTS: Protease and RT sequences were successfully amplified from only 37 (35.6%) of the 104 seroconverters. Only one patient who seroconverted in August 1991 showed any evidence of significant mutations in the RT region, and this was associated with resistance to zidovudine (ZDV) (215Y and 210W). An additional patient who seroconverted in July 1991 had a TOR mutation and was classified as having intermediate resistance to ZDV. No spontaneous mutations were detected in the protease region. CONCLUSIONS: Overall only 2 (5%) of these treatment-naive individuals were infected with HIV variants resistant to ZDV. Although the data at present do not support the need for pretreatment genotyping, there is a need for continued surveillance of the frequency of resistance mutations in antiretroviral naive patients since the introduction of highly active antiretroviral therapy. PMID- 12076069 TI - Meningococcal disease presenting as bronchiolitis. AB - Meningococcal disease is the leading infective cause of mortality in children. Bronchiolitis and meningococcal disease share some common features. Both are seasonal diseases with epidemics in winter. A preceding history of upper respiratory tract infection is commonly present in both. We report two cases of meningococcal disease in infants whose initial presentation was suggestive of bronchiolitis. We draw attention to tachypnoea as an important but overlooked early sign of meningococcal septicaemia. PMID- 12076070 TI - A case of lower respiratory tract infection caused by Neisseria weaveri and review of the literature. AB - Neisseria weaveri (formerly CDC [Centers for Disease Control and Prevention] group M-5 is part of the normal canine oral flora. Infections in humans are usually associated with dog bite wounds. Very rarely the organism has been isolated from sites other than wounds, or from deep seated infections. A 60-year old man was admitted to our hospital because of an acute exacerbation of his bronchiectasis. Gram stain of bronchial washings and expectorated sputum showed numerous polymorphs and Gram-negative bacilli. Routine bacterial culture yielded a heavy pure growth of a Gram-negative rod-shaped organism that was strongly oxidase and catalase positive, indole negative, non-motile and did not ferment carbohydrates. The organism was identified as N. weaveri by using 16S rRNA sequencing. The patient was treated with a 3 weeks course of ofloxacin and had a good response. Sputum culture after treatment yielded normal respiratory flora only. To our knowledge, this is the first reported case of lower respiratory tract infection caused by N. weaveri. PMID- 12076071 TI - Chorioamnionitis and neonatal septicaemia caused by Eikenella corrodens. PMID- 12076072 TI - Congenital and maternal cytomegalovirus infection in a north west population. PMID- 12076073 TI - Acute acalculous cholecystitis associated with co-infection by Plasmodium falciparum and Plasmodium vivax. PMID- 12076074 TI - Hospital-acquired, native valve endocarditis caused by Pseudomonas aeruginosa. PMID- 12076075 TI - A change in pattern of group G streptococcal septicaemia? PMID- 12076076 TI - Somatostatin analogs inhibit neonatal retinal neovascularization. AB - The goal of this study was to determine the effect of two somatostatin analogs, Woc4D and octreotide, on oxygen induced retinopathy in the mouse. Oxygen induced retinopathy was produced in C57BL6 mice. Octreotide and Woc4D were administered from post-natal day 12-16. Retinopathy was assessed by a retinal scoring system utilizing fluorescein perfused retinal whole mounts. Animals treated with Woc4D and octreotide, respectively, had median retinopathy scores of 4(3,5) [median(25th, 75th quartile)] with P = 0.01 and 3.5(2.9,4.3) with P = 0.01 compared to oxygen and sham treated oxygen animals with scores of 6.6(5.3,8.5) and 7.4(5.8,8.6), respectively. Woc4D and octreotide treated animals had decreased blood vessel tufts and decreased extra-retinal neovascularization when compared to oxygen treated animals. Pituitary growth hormone (GH) mRNA expression was increased 8.3-fold by Woc4D treatment and 106-fold by oxygen exposure, and GH and mRNA was markedly reduced by Woc4D as well as octreotide. Growth as measured by animal weight was unaffected by either treatment. Woc4D and octreotide inhibited retinal neovascularization in an equally effective manner in the mouse model of oxygen induced retinopathy. PMID- 12076077 TI - Temporal relationship of choroidal blood flow and thickness changes during recovery from form deprivation myopia in chicks. AB - When form deprived, young chicks rapidly develop axial myopia, from which they recover if the treatment is ceased at a sufficiently early age. The increased axial growth of the eye is accompanied by choroidal thinning and decreased choroidal blood flow (ChBF). In contrast, during the early part of the recovery process, the choroid thickens, shifting the retina towards the new plane of focus. Little information is available about ChBF during recovery from myopia. Because of the possibility that choroidal thickening during recovery from myopia might be driven by an increase in ChBF, the temporal relationship of ChBF and choroidal thickness changes was examined during such recovery. White Leghorn chicks were form deprived from 3 days of age for 2-3 weeks using detachable plastic diffusers. Axial ocular dimensions, including choroidal thickness, were then measured by high frequency A-scan ultrasonography at various times after the diffusers were removed up to 240 hr. ChBF was measured transclerally immediately following the A-scan ultrasonography, using laser Doppler flowmetry. In the chicks measured immediately after diffuser removal, the vitreous chamber was 29.9% longer, the choroid was 6.4% thinner and ChBF was 13.7% less in the treated than in the non-treated control eyes. These changes are characteristic of myopic chick eyes and are reversible in young eyes. Thus, in chicks examined 7 hr after diffuser removal, the ChBF in recovering eyes was now greater than that in control eyes. This ChBF increase peaked about 19 hr after the diffusers were removed. The mean increase in ChBF in treated eyes for the 7-30 hr monitoring period was 187%, relative to control eyes. ChBF in the treated eyes gradually returned to the control level after this time. By contrast to the early, transient increase in ChBF, significant choroidal thickening was not observed in treated eyes until 30 hr after diffuser removal, and continued to increase relative to control eyes over the remainder of the monitoring period, reaching a final mean value of 182%. This study demonstrates, in chick eyes recovering from form deprivation myopia, large increases in ChBF that preceded increases in choroidal thickness and were also more transient than the latter. These results raise the possibility that the increase in ChBF may trigger or even drive the subsequent onset of choroidal expansion, perhaps by facilitating the filling of the choroidal lymphatic lacunae that are well developed in the avian eye. PMID- 12076078 TI - Cultivation of retinal pigment epithelial cells from human choroidal neovascular membranes in age related macular degeneration. AB - A method is described for cultivating retinal pigment epithelial cells from choroidal neovascular membrane (CNV) specimens that were surgically removed in patients with age-related macular degeneration (AMD). CNV specimens of 43 patients were available for cultivation. They were incubated in supplemented DMEM/Ham's F12 cell culture medium on microporous semipermeable filter membranes. Thirty-four specimens gave rise to cell cultures, 28 of which could be subcultivated for up to 15 passages. The membrane type as classified by fluorescence angiography was compared with cellular growth in vitro. Immunocytochemistry revealed a uniform expression of cytokeratin 18 and vimentin, while factor 8, glial fibrillary acidic protein and alpha smooth muscle actin were absent in all 21 cultures stained. The expression of RPE markers cellular retinaldehyde binding protein (CRALBP) and RPE65 was detected by RT-PCR in all cultures tested. An epithelial character of the cultures was supported by the presence of apical microvilli as determined by electron microscopical studies. Therefore, the cell cultures from CNV in AMD bear characteristics of retinal pigment epithelial cells. For the first time, this cell culture system holds the potential to study human RPE cells in the context of neovascular AMD in vitro. PMID- 12076079 TI - Expression of matrix metalloproteinases and their inhibitors in experimental retinal ischemia-reperfusion injury in rats. AB - Matrix metalloproteinases (MMPs) are endopeptidases that degrade the extracellular matrix (ECM) and are involved in the pathogenesis of retinal degeneration along with tissue inhibitors of metalloproteinases (TIMPs). The present study examined the expression and activation of two specific members of MMPs (MMP-2 and MMP-9) and their related inhibitors (TIMP-1 and TIMP-2) in an experimental retinal ischemia-reperfusion injury. Retinal ischemia-reperfusion injury (RIRI) was induced in adult rats with a ligation method. After one hour of ischemia and a varied reperfusion time (0, 3, 6, 12, 24, 48 and 76 hr), the rat eyes were enucleated. Retinal extracts underwent zymographic analysis to measure the activity of MMP-2/9. The activity of TIMP-1 and TIMP-2 was measured by reverse zymography. The protein level was examined by Western blot. Immunohistochemistry analysis was undertaken to assess the anatomical distribution of MMP-9 in the retina after RIRI. The gelatinolytic activity of ProMMP-2 (72 kDa) was increased markedly at 6 hr after RIRI. ProMMP-9 (92 kDa) was not detected in the control specimens, while it appeared at 3 hr, increased markedly at 6 hr, and reached maximal levels at 24 hr after RIRI. The gelatinolytic activity found ian retinal extracts was shown to be inhibited by 10 m M EDTA and activated in vitro by a known metalloproteinase activator (4 aminophenylmercuric acetate (APMA)), indicating that these enzymes were of the metalloproteinase class. By western blot, MMP-2/9 levels increased parallel to protein activity level in zymography. No corresponding increase in TIMP-1 and TIMP-2 protein activity and protein level was detected by reverse zymography and western blot. Elevated levels of MMP-9 and its distribution in retina were confirmed by immunohistochemistry. Expression of MMP-9 was detected in the inner and outer segments of rat retina, and the level becomes stronger at 24 hr after RIRI. In this study, ProMMP-2 and ProMMP-9 were expressed and increased significantly, but their inhibitors (TIMP-1 and TIMP-2) remained relatively unaltered in ischemic retina after RIRI in rats. These results suggest that MMP-2 and MMP-9 may play an important role in the pathomechanism of retinal ischemic injury. PMID- 12076080 TI - Inhibitory effects of salmosin, a disintegrin, on posterior capsular opacification in vitro and in vivo. AB - The proliferation, migration and transdifferentiation of the remaining lens epithelial cells (LECs) after cataract surgery are a major cause of posterior capsular opacification (PCO). It has previously been reported that salmosin, a novel disintegrin, significantly inhibits solid tumor growth in mice by perturbation of tumor-specific angiogenesis via blocking alpha v beta 3 integrin expressed on vascular endothelial cells. In this study, the inhibitory function of salmosin in PCO was investigated and was found that salmosin inhibits the attachment of bovine LECs and rabbit lens cells (N/N1003A) to extracellular matrix-coated plates. The anti-adhesive activity of salmosin was approximately 1000 times higher than that of synthetic Arg-Gly-Asp peptide. In addition, the cell proliferation and migration of bovine LECs and N/N1003A were strongly inhibited by salmosin, whereas the proliferation of corneal endothelial cells was less affected. LEC migration and proliferation were also decreased by salmosin treatment in rabbit eyes without any toxic effect in the cornea, iris and retina. In this study, salmosin was shown to specifically inhibit LEC migration and proliferation in an animal model. Therefore, the authors suggest that further investigation may show salmosin to be a good candidate for inhibiting PCO development. PMID- 12076081 TI - Changes in three types of ubiquitin mRNA and ubiquitin-protein conjugate levels during lens development. AB - Ubiquitin is a small, highly conserved protein that covalently attaches to other proteins to form a unique branched protein structure. The best characterized function of this post-translational modification is to mark the modified protein for degradation by the proteasome. To investigate whether ubiquitin genes are regulated in lens development, the authors analyzed the levels of three ubiquitin mRNAs (UbA(52), UbB and UbC) in freshly dissected fiber and epithelial cells, and in epithelial explants induced to differentiate ex vivo. Explants, comprising the capsule and adherent epithelial cells, were dissected from lenses of 3 day old Sprague Dawley rats and cultured +/-bFGF to induce differentiation. Quantitative competitive RT-PCR was used to determine the mRNA levels in fresh and cultured cells. UbA(52), UbB and UbC mRNAs were 3.2 (P < 0.0001), 5.0 (P < 0.0001) and 6.8 (P < 0.0001) fold higher, respectively, in freshly dissected epithelial cells than in differentiated fiber cells. Immunological spot assays showed that ubiquitin protein is over two fold as high in rat pup lens epithelial cells as in fiber cells. The ubiquitin protein in fiber cells of adult rat is lower than that in adult epithelium and in pup fiber cells, indicating that ubiquitin content further decreased during lens fiber maturation. Western blots showed a greater amount of protein-conjugated ubiquitin (MW > 81 kD) in epithelial cells than in fiber cells, demonstrating a parallel pattern between the expression of ubiquitin mRNA, the level of ubiquitin protein and the level of conjugates in the cells. Epithelial cell explant cultures permit study of cells initiating differentiation. In contrast to fully differentiated fiber cells, explant cultures induced to initiate differentiation underwent differential up-regulation of ubiquitin gene expression. UbA(52) and UbB mRNA levels in +bFGF (differentiating) explant cultures were 2.6 (P < 0.001) and 1.4 (P < 0.001) fold higher, respectively, than those of -bFGF cultures. UbC mRNA content was similar in explants cultured with or without bFGF. Dissection of the isolated epithelial cells into regions representing distinct populations gave results consistent with this observation of the explant results. UbA(52), UbB and UbC mRNAs are 2.0, 2.2 and 1.76 fold higher, respectively, in the peripheral (initiating differentiation) than in the central (undifferentiated) region of epithelial cells. These results together indicate that UbA(52) and UbB mRNAs are transiently increased during the initiation and early stages of differentiation. However, UbC mRNA appears to be relatively unaffected at the earliest stage in this differentiation model and may have a different distribution than UbA(52) and UbB in the anterior lens cells. These data are consistent with an important role for ubiquitin during the early stages of lens differentiation. The selective expression indicates that the three genes have specific differentiation related functions. PMID- 12076082 TI - Expression of spermidine/spermine N(1)-acetyltransferase in the Muller glial cells of the developing chick retina. AB - A number of genes have been found to be asymmetrically expressed along the three axes of the retina: central-peripheral, dorsal-ventral, temporal-nasal. Some of the asymmetrically expressed genes have been shown to play a role in the establishment of boundaries required for guiding retinal axons to their correct targets in the brain. Asymmetric expression during development can also be a consequence of the different rates of differentiation along the three retinal axes. The authors have used a differential-display-PCR approach to identify genes asymmetrically expressed along the dorsal-ventral axis in the chick retina. One of the selected genes, spermidine/spermine N(1)-acetyltransferase (SSAT), was preferentially expressed in the dorsal-temporal quadrant of the developing retina. There was a sharp increase in retinal SSAT mRNA levels during the transition stage from proliferation (E7) to early differentiation (E10). SSAT mRNA was found in Muller glial cells and its distribution pattern in these cells closely followed the three differentiation axes of the developing retina, with a central-dorsal-temporal preference. The elevated levels of SSAT mRNA in Muller glial cells may reflect a requirement for acetylated spermine/spermidine or putrescine in the differentiating neuronal cells of the retina. PMID- 12076083 TI - Early retinal damage in experimental diabetes: electroretinographical and morphological observations. AB - A growing body of evidence indicates that impairment of retinal function precedes the earliest signs of vascular complications. The aim of this study was to follow the development of retinopathy both functionally and morphologically in a rat model of diabetes mellitus. Diabetes was induced in rats by intravenous injection of streptozotocin (STZ). Age-matched rats raised under similar conditions served as control. The electroretinogram (ERG) was recorded in order to assess retinal function. The expression of glial fibrillary acidic protein (GFAP) in Muller cells was used as a cellular marker for retinal damage. The ERG responses of the diabetic rats were reduced in amplitude compared to the responses recorded from the control rats as early as 2 weeks after onset of diabetes. The b-wave was more affected than the a-wave. GFAP expression in the diabetic retina did not differ from that in the control retina during the first 5 weeks of diabetes. GFAP was demonstrated only in astrocytes in the vitreo-retinal border. After 6-7 weeks of diabetes, GFAP expression in the retinas of the diabetic rats was also detected in the endfeet of the Muller cells. With the progression of diabetes, GFAP expression spreads throughout the entire length of the Muller cells. In the retinas from control rats, GFAP expression was limited to astrocytes and was not detected in Muller cells even at 40 weeks of follow-up. The observations indicate that the functional integrity of retinal cells is compromised already at short time intervals after onset of experimental diabetes in rats. PMID- 12076084 TI - Human retinal pigment epithelial cells inhibit proliferation and IL2R expression of activated T cells. AB - The purpose of this study was to characterize the effects of human retinal pigment epithelial (RPE) cells on activated T cells. Activated T cells were cocultured with adult and foetal human RPE cells whereafter apoptosis and proliferation were determined by flow cytometry and (3)H-Thymidine incorporation assay, respectively. T cells and RPE cells were cultured directly together or in a transwell system for determination of the effect of cell contact. The importance of cell surface molecules was examined by application of a panel of blocking antibodies (CD2, CD18, CD40, CD40L, CD54, CD58) in addition to use of TCR negative T cell lines. The expression of IL2R-alpha -beta and -gamma chains of activated T cells was analysed by flow cytometry after incubation of T cells alone or with RPE cells. Human RPE cells were found to inhibit the proliferation of activated T cells by a cell contact-dependent mechanism. The RPE cells inhibitory abilities were not affected by blocking of any of the tested surface molecules. The inhibition of the T cells' proliferation correlates with a decreased expression of IL2R-beta and -gamma chains. The T cells regain their ability to proliferate and increase their IL2R-beta and -gamma chain expression within 24 hr after removal from the coculture. It is concluded that the cultured human adult and foetal RPE cells inhibit the proliferation of activated T cells by a process that does not involve apoptosis. It depends on cell contact but the involved surface molecules were not revealed. The proliferation inhibition correlates with a modulation of the T cells' expression of IL2R, and is reversible. PMID- 12076085 TI - Delivery of genes and fluorescent dyes into cells of the intact lens by particle bombardment. AB - The authors report the use of a particle bombardment technique to deliver exogenous genes and fluorescent dyes into living fibre cells in the intact lens. Gold particles were coated with plasmid DNA encoding green fluorescent protein (GFP) or the lipophilic fluorescent probes DiI and DiO. The particles were introduced into embryonic chicken or neonatal mouse lenses using a pressurized helium charge. A significant fraction of particles penetrated the capsule and came to rest in the superficial lens cortex. Over the range tested, varying the particle size or the pressure of the helium charge had little effect on the final distribution of particles within the lens. Particle bombardment was used initially to deliver DiI and DiO into the lens. Within a few hours, bombarded lenses exhibited multicolored membranous labelling of individual, elongating fibre cells. The particle bombardment technique was also used to introduce a plasmid encoding GFP. After overnight incubation, many fibre cells in the bow region expressed GFP. On close examination by confocal reflectance microscopy, each expressing cell was found to contain a gold particle lodged in its nucleus. The authors examined the fate of GFP-expressing fibre cells over a period of 1 week in organ culture. In the embryonic chicken lens, transfected fibres showed modest (approximately two-fold) elongation. In contrast, GFP-expressing mouse lens fibres underwent dramatic elongation, reaching the anterior and posterior sutures after 7 days in culture. These species differences may reflect the fact that mitosis continued at a near normal rate in the cultured mouse lens but declined precipitously in the cultured chicken lens. These results suggest that particle bombardment, in conjunction with appropriate cell culture conditions, may prove useful in visualizing the behaviour of differentiating fibre cells in the living intact lens in vitro. PMID- 12076086 TI - Up-regulation of cytochrome oxidase in the retina following optic nerve injury. AB - The purpose of this study is to investigate the cytochrome oxidase (COX) activity in the retina and optic nerve following an optic nerve injury. The optic nerve crush of one eye was carried out in Balb/c mice. A semi-quantitative RT-PCR method was then adopted to evaluate the mRNA expression of cytochrome oxidase subunit 1 (COX1) in the retina after surgery. Up-regulation of COX1 mRNA in the retina was detected by RT-PCR at 24 hr following the optic nerve injury. Total retinal mitochondrial mass measured by fluorescent intensity of MitoTracker green was not altered following the injury. COX histochemistry performed on cryostat sections showed an elevated enzyme activity of COX in the retina and in the optic nerve. In the retina, elevation of the COX activity was observed in the retinal ganglion cell layer and the overlying nerve fibre layer. The increase of COX activity began from 24 hr after injury, peaked around day 3, and maintained up to 1 week after the operation. In the optic nerve, increase of COX activity was observed in regions distal to the crush line and distributed either randomly or in a cone shape. In conclusion, both the expression of COX1 mRNA in retina and the activity of COX in inner plexiform layer and retinal ganglion cell layer were elevated following optic nerve injury without affecting total retinal mitochondrial mass. These findings suggested that one of early responses in the retina and in the optic nerve after the optic nerve injury is to scale up the energy production. PMID- 12076087 TI - Distribution and density of medium- and short-wavelength selective cones in the domestic pig retina. AB - The topography of medium (M)- and short (S)-wavelength sensitive cone photoreceptors was studied in the domestic pig retina. Antisera specific for M or S opsin as well as cone photoreceptor proteins arrestin and alpha-transducin were used to label cone types. Retinal wholemounts and their blood vessel patterns were drawn and specific regions removed. The wholemounts were immunocytochemically labelled to detect both M and S cones, and the specific regions labelled to detect S cones. Cones were counted in a 1 mm grid pattern, using the drawings as a guide. Pig retina has a high cone density retinal streak extending across the retina covering the optic disc (OD) and horizontal meridian. Densities in the streak are 20,000-35,000 mm(-2). Two higher peaks occur in the streak, one in temporal retina near the OD (39,000 mm(-2)) and the other in nasal retina 5-7 mm from the OD (40,500 mm(-2)). The lowest cone density is in far peripheral inferior retina (7000 mm(-2)). The total number of cones in pig retina is 17-20 million. Both types of cones are found throughout the retina, with S cone percentages ranging from 7.4 to 17.5% in no consistent topographical pattern. S cones have an irregular local distribution which can vary from a regular hexagonal pattern to small clusters of adjacent S cones to small areas lacking S cones. Double-label immunocytochemistry found that virtually all S cone outer segments (OS) contain some M opsin. M cone OS do not label at detectible levels for S opsin. Domestic pig retina is widely available, large, has a high cone density and has two types of cones. This tissue should be an excellent source for biochemical analysis of cone proteins, and for in vitro approaches to understanding cone survival factors. PMID- 12076088 TI - Levobetaxolol-induced Up-regulation of retinal bFGF and CNTF mRNAs and preservation of retinal function against a photic-induced retinopathy. AB - Betaxolol (racemic), a beta-adrenoceptor antagonist that is used to lower intraocular pressure in the treatment of glaucoma, has been shown to protect inner retina cells from various insults. To determine if such protection could be afforded to retinal photoreceptors and retinal pigment epithelial cells (RPE), levobetaxolol (S-betaxolol) was evaluated in a photic-induced retinopathy model. Rats were dosed (IP) with vehicle or levobetaxolol (10 and 20 mg kg(-1)) 48, 24 and 0 hr prior to exposure for 6 hr to fluorescent blue light. The electroretinogram (ERG) and retinal morphology were assessed after a 3 week recovery period. Evaluation of the ERG demonstrated significant protection of retinal function in levobetaxolol (20 mg kg(-1))-dosed rats compared to vehicle dosed rats. Similarly, the RPE and outer nuclear layer were significantly thicker in levobetaxolol (20 mg kg(-1))-dosed rats compared to vehicle-dosed rats. To elucidate potential mechanism(s) of the neuroprotective activity of levobetaxolol, bFGF and CNTF mRNA levels in normal rat retinas were evaluated 12 hr after a single i.p. injection. Northern blot analysis of levobetaxolol treated retinas demonstrated a 10-fold up-regulation of bFGF and a two-fold up-regulation of CNTF mRNA levels, trophic factors that have been shown to inhibit retinal degeneration in a number of species. These studies suggest that levobetaxolol can be used as a novel neuroprotective agent to ameliorate retinopathy. PMID- 12076089 TI - Genetic modification of retinal degeneration in tubby mice. AB - Mice that carry the recessive mutation tub develop neurosensory defects including retinal and cochlear degeneration, as well as maturity-onset obesity associated with insulin resistance. The biological function of the gene and the mechanism by which it induces its phenotypes are still unclear. In order to elucidate the pathways through which tub functions, in the current study, QTL modifiers were identified in an F2 intercross between (C57BL/6J- tub/tub and AKR/J-+/+) F1 hybrids (AKR intercross). The thickness of the outer nuclear layer of the retina and the number of photoreceptor nuclei were assessed in F2 mice homozygous for the tub mutation. A genome-wide scan revealed a significant linkage on chromosome 11 (named motr1) and two suggestive linkages on chromosomes 2 and 8. Interestingly, the same chromosome 2 region identified for the hearing modifier of tubby, the moth1 locus, showed a peak lod score of 2.3 for protection from retinal degeneration. This result suggests that the gene responsible for the QTL on chromosome 2 might be involved in a common pathway through which retinal and cochlear degeneration are induced in tubby mice. PMID- 12076090 TI - Nitration of manganese superoxide dismutase during ocular inflammation. AB - Reactive nitrogen species, in particular, peroxynitrite (ONOO(-)) have been proposed to play an important role in the pathogenesis of endotoxin-induced uveitis (EIU). Tyrosine nitration by ONOO(-) has been shown in other model systems to inhibit the activity of the superoxide anion quenching enyzme, manganese superoxide dismutase (MnSOD), perhaps contributing to progression of disease. In this study, it is confirmed through immunoanalysis that nitrated proteins are produced during EIU, and furthermore, that MnSOD is a target of nitration during the inflammatory response. In addition, through microsequencing analyses, nitrated albumin--apparent in both control and EIU eyes--was identified. Positive immunostaining of nitrated proteins was seen in the ciliary epithelium, inflammatory cells, and protein exudate of eyes from rats injected with endotoxin. Incubation of nitrotyrosine immunoprecipitates from the iris and ciliary body (ICB) with a polyclonal antibody against MnSOD revealed that nitrated MnSOD was present only in the ICB of EIU rats. When the total activity of the enzyme was examined, it was observed that despite the presence of nitrated MnSOD, activity was increased relative to control. Analysis of MnSOD mRNA and protein from the ICB of both groups demonstrated an increase in mRNA expression and consequently a three- to five-fold increase in MnSOD protein in EIU rats as compared to control rats. Further examination of MnSOD protein expression through immunohistochemistry noted enhanced immunostaining in the ciliary epithelium of eyes of EIU rats. Additional investigation of a 70 kDa band apparent in nitrotyrosine immunoprecipitates from the ICB of control and EIU rats revealed that the plasma protein albumin is nitrated as well. This protein is present as a result of the breakdown of the blood-aqueous barrier during inflammation. In summary, two endogenous nitration targets, albumin and MnSOD, were identified. Nitrated MnSOD appears to be specifically targeted to the ICB during inflammation, underscoring the importance of the interface in EIU. Furthermore, the expression and activity of the enzyme is increased in the ICB during EIU, perhaps regulating reactive nitrogen species produced within the cells. This study implicates ONOO(-) in the pathogenesis of EIU and imparts the putative role MnSOD plays in disease resolution. PMID- 12076091 TI - Cellular incursion into Bowman's membrane in the peripheral cone of the keratoconic cornea. AB - Analysis of corneal tissue from normal and keratoconic donors has revealed differences which may represent early signs in the pathogenesis of keratoconus. Peripheral areas of keratoconic tissue obtained from transplant surgery were targeted to ascertain cellular disposition and morphological changes which may be masked within the extensive damage of the central keratoconic cone. Peripheral keratoconic corneae exhibited discrete incursion of fine cellular processes into Bowman's membrane. These processes originated from keratocytes and were often observed in conjunction with a defined indentation from the basal epithelium. Comparison of the lysosomal enzymes cathepsin B and G with constitutively expressed cytoplasmic esterase determined that both cathepsins were elevated within keratocytes of keratoconic tissue compared with normal tissue. Some clusters of keratoconic keratocytes had elevated levels of cathepsin exceeding all others. Cathepsin-rich keratocytes localized with morphologically compromised regions of Bowman's membrane. The presence of cell nests deeper within the stroma indicated that the catabolic changes, which are visible within the acellular Bowman's membrane, are probably also occurring deeper within the stroma, but are masked and not readily detectable. PMID- 12076092 TI - Evaluation of therapeutic effectiveness using MR imaging in a rabbit model of anterior uveitis. AB - Magnetic resonance imaging (MRI) has been used to examine conditions that alter the permeability of the blood-retinal barrier. Our goal was to determine if blood aqueous barrier permeability could be similarly assessed, because MRI offers the theoretical advantage of providing quantitative data directly from inflamed uveal tissues rather than from the aqueous humor into which the inflammatory reaction spills. As an additional challenge, we sought to use MRI to measure differences between the inflamed uveal tissues of corticosteroid-treated and placebo-treated uveitic eyes. Anterior uveitis was induced in one eye of eight rabbits by subcutaneous injection of Mycobacterium tuberculosis, followed after 10 days with intravitreal challenge. One rabbit of each pair was treated with topical 1% prednisolone acetate while control rabbits were treated with artificial tears. Contrast-enhanced MRI studies were performed prior to uveitis induction, one day after induction and then weekly for at least 2 weeks. MR image data were analyzed to determine percent change in peak enhancement of the ciliary body and anterior chamber. The initial rate of change of enhancement of the anterior chamber was also measured. Extensive contrast agent-induced MR image enhancement of both the anterior chamber and the ciliary processes was measured following the induction of uveitis. More rapid improvement was measured for the 1% prednisolone acetate treated rabbit eyes (P < 0.001). MR signal enhancement data obtained from the ciliary processes proved to be the most reliable indicator of disease activity in this rabbit model of uveitis. Such data can only be obtained using MRI. PMID- 12076093 TI - Ischemia-reperfusion injury of retinal endothelium by cyclooxygenase- and xanthine oxidase-derived superoxide. AB - The formation of reactive oxygen species (ROS) may be important in the pathogenesis of microvascular dysfunction and injury in ischemic retinopathies. The authors hypothesized that retinal endothelial cells can generate injurious levels of superoxide radical in response to ischemia/reperfusion, that endothelial xanthine oxidase and cyclooxygenase are important enzymatic sources of superoxide radical under these conditions, and that superoxide scavengers and inhibitors of these enzymes can protect endothelium from ischemic injury. The authors used confluent cultures of mouse retinal endothelial cells (MREC) subjected to exogenously generated superoxide or simulated ischemia-reperfusion to test these hypotheses. Cell injury was assessed biochemically by lactate dehydrogenase release into the culture medium. MREC were injured in a duration dependent fashion by exposure to the superoxide-generating mix of hypoxanthine and xanthine oxidase. Increasing periods of oxygen and glucose deprivation (OGD) for 5-9 hr followed by replenishment of substrates for 2 hr led to progressive increases in endothelial cell injury; a significant proportion of the injury occurred during the period of substrate replenishment. Significant MREC protection was achieved by the superoxide scavengers SOD (1000 U ml(-1)) and a carboxylic acid derivative of carboxyfullerene (10 microM), the xanthine oxidase inhibitors oxypurinol (100 microM) and diphenyleneiodonium (DPI) (100 n M), and the cyclooxygenase inhibitors indomethacin (300 microM) and ibuprofen (300 microM). It is concluded that MREC are vulnerable to auto-oxidative injury by superoxide radical generated following a period of OGD. Both xanthine oxidase- and cyclooxygenase-dependent pathways are important enzymatic sources of superoxide formation in this setting. These enzymes and the ROS produced from their activity may be viable therapeutic targets to reduce microvascular dysfunction and injury in ischemic retinopathies. PMID- 12076094 TI - Glutathione and NADH, but not ascorbate, protect lens proteins from modification by UV filters. AB - Age-dependent human lens colouration and fluorescence may stem primarily from the covalent binding of UV filters to crystallins. The tendency of the kynurenine (Kyn) UV filters to deaminate at neutral pH, with the generation of reactive alpha,beta-ketoalkenes, underlies this phenomenon. In this study the authors examined the ability of small molecular weight antioxidants, which are known to be present in the lens, to inhibit this process. Crystallins were incubated with Kyn at pH 7 in the presence of glutathione (GSH), ascorbate or NADH. Ascorbate, even at high (15 m M) levels, was not found to significantly retard the time dependent covalent binding of Kyn to the proteins. GSH, and to a lesser extent NADH, however, had a major impact in preventing this modification. The increase in protein UV absorbance and fluorescence was inhibited by GSH intercepting the reactive ketone intermediate, to form a GSH-Kyn adduct. NADH seemed to protect by both reduction of the reactive ketone intermediate and by competing with Kyn for presumably hydrophobic sites on the crystallins. This may indicate that the covalent attachment of aromatic Kyn molecules could be facilitated by initial hydrophobic interactions. Since GSH is present at far greater concentrations than NADH, these results show that in primate lenses, GSH is the key agent responsible for protecting the crystallins from covalent modification. PMID- 12076095 TI - Retinal pigment epithelial cell DNA is damaged by exposure to benzo[a]pyrene, a constituent of cigarette smoke. AB - This study examined the effect of exogenous benzo[ a ]pyrene (BaP), an important constituent of cigarette smoke, on cultured bovine retinal pigment epithelial (RPE) cells. Evidence is presented for its metabolic conversion into benzo[ a ]pyrene diol epoxide (BPDE) and the consequent formation of potentially cytotoxic nucleobase adducts in DNA. Cultured RPE cells were treated with BaP at concentrations in the range of 0-100 microM. The presence of BaP was found to cause inhibition of cell growth and replication. BaP induced the expression of a phase I drug metabolizing enzyme which was identified as cytochrome P450 1A1 (CYP 1A1) by RT-PCR and by Western blotting. Coincident with the increased expression of CYP 1A1, covalent adducts between the mutagenic metabolite BPDE and DNA could be detected within RPE cells by immunocytochemical staining. Additional support for their formation was afforded by nuclease P1 enhanced (32)P-postlabelling assays on cellular DNA. Single-cell gel electrophoresis (comet) assays showed that exposure of RPE cells to BaP rendered them markedly more susceptible to DNA damage induced by broad band UVB or blue light laser irradiation. In the case of UVB, this is consistent with the photosensitization of DNA cleavage by nucleobase adducts of BPDE. Collectively, these findings imply that BaP has a significant impact on RPE cell pathophysiology and suggest mechanisms whereby exposure to cigarette smoke might cause RPE dysfunction and cell death, thus possibly contributing to degenerative disorders of the retina. PMID- 12076097 TI - Expansion of conjunctival epithelial progenitor cells on amniotic membrane. AB - Amniotic membrane (AM) reconstructed human conjunctival surfaces recover a goblet cell density higher than normal. Cultured rabbit conjunctival epithelial cells (RCE) on AM preferentially exhibit non-goblet epithelial differentiation. It was thus wondered if conjunctival progenitor cells that might have been preserved during ex vivo expansion on AM can still differentiate into conjunctival non goblet epithelial and goblet cells under the influence of mesenchymal cells. Fourteen day old AM cultures of RCE were subcutaneously implanted in Balb/c athymic mice for 11 days and processed for PAS staining and immunostaining with monoclonal antibodies to conjunctival goblet cell mucin (MUC5AC, AM3), glycocalyx (AMEM2), cornea specific cytokeratins K3 (AE5) and K12 (AK2) and basal cell specific cytokeratin K14. Cell cycle kinetics were measured by BrdU labelling for 1 or 7 days. The 7 day labelled RCE were chased for 14 days in the same primary culture. After subcutaneous implantation, conjunctival non-goblet epithelial cells increased stratification and formed occasional cysts. The resultant epithelial phenotype was conjunctival with many PAS-positive, MUC5AC-positive, and AM3-positive goblet cells, AMEM2-positive suprabasal and superficial cells, and K14-positive basal cells, but was not corneal (negative to AE5 and AK2 staining). Twenty four hr BrdU labelling showed a labelling index of 42.5%. A higher labelling index or 69% was noted after continuous BrdU labelling for 7 days. A large number of label retaining basal cells with a labelling index of 84% were noted following 14 days of chase. Conjunctival epithelial progenitor cells for goblet and non-goblet cell differentiation are preserved by AM in vitro as evidenced by being able to differentiate into goblet cells in a permissive stromal environment, and being slow-cycling, and label retaining. This information is useful for future ex vivo expansion of conjunctival epithelial stem cells for conjunctival surface reconstruction. PMID- 12076096 TI - Effects of angiogenic growth factor combinations on retinal endothelial cells. AB - The aim of this paper was to determine if growth factors, known to be upregulated in proliferative diabetic retinopathy, exerted combined effects on retinal endothelial cells. The authors explored the individual and collective actions of insulin-like growth factor I (IGF-I), vascular endothelial growth factor (VEGF), platelet-derived growth factor-BB (PDGF-BB), fibroblast growth factor-2 (FGF-2) and placenta growth factor (PlGF) on several parameters that reflect the angiogenic potential of endothelial cells. The effect of growth factors on cell migration and survival/proliferation was examined using primary cultures of bovine retinal endothelial cells (BREC). The authors also determined the growth factor action on capillary-like tube formation on a reconstituted basement membrane matrix and on the newly described phenomenon of secondary sprouting, in which endothelial cell colonies spontaneously survive, proliferate, migrate and invade the matrix after the original capillary-like tubes have collapsed. Sprouting cells were positive for von Willebrand factor and could aggregate into larger tubes with lumens. Incubation with VEGF+IGF-I or PlGF+FGF-2 enhanced tube stability by 40-50%, more than each growth factor alone or other combinations (5 20%). The concurrent addition of four growth factors did not improve the response seen with growth factor pairs. Surprisingly, PDGF-BB induced tube collapse. IGF-I and FGF-2 mildly enhanced BREC proliferation/survival (5-15%). However, VEGF+IGF I or PlGF+FGF-2 increased BREC proliferation/survival by 25% under low serum conditions, whereas combinations of all four growth factors exerted a clearly synergistic effect (250% increase). PDGF-BB or FGF-2 stimulated secondary sprouting and were the only factors capable of exerting this effect alone. Even though VEGF, IGF-I or PlGF were not effective, if administered in pairs, they demonstrated increased responses. PDGF-BB was also able to enhance the effect of FGF-2+IGF-I+VEGF on BREC secondary sprouting, but not of any of them individually. No other growth factor tested was able to significantly improve the action of combinations of three other growth factors. VEGF increased cell migration in a wounded monolayer assay two-fold and PDGF-BB, 2.5 times, but other individual growth factors were ineffective. PlGF+FGF-2 enhanced cell migration more than each factor alone. VEGF+IGF-I+PlGF+FGF-2, however, increased cell migration four-fold. In summary, this study indicates that growth factors, overexpressed in diabetic retinopathy eyes, enhance the angiogenic characteristics of cultured cells (tube formation, proliferation, secondary sprouting and migration). Their effects, however, can be greatly augmented by other growth factors that alone exert little or no action. Therefore, diabetic retinal neovascularization may result from the additive or synergistic action of several growth factors. PMID- 12076098 TI - Clusterin is present in drusen in age-related macular degeneration. PMID- 12076100 TI - Application of semiological seizure classification to epileptic seizures in children. AB - To better define seizure characteristics and to examine whether semiological seizure classification (SSC) can be appropriately applied to the seizures of infants and children, we studied 152 videotaped seizures recorded in 133 children. Seizure semiology was described on the basis of a series of behavioural, sensory, and motor phenomena according to SSC. Special emphasis was placed on whether one semiology can be representative of a patient's whole semiology sequence. If one semiology was able to represent the whole sequence, the seizure was classified according to SSC. Ninety of 152 seizures (59.2%) could be classified as a single seizure type by SSC. However, only 19 of 53 seizures (35.9%) consisting of two semiologies, three of 24 seizures (12.5%) consisting of three semiologies, and one of eight seizures (12.5%) consisting of four semiologies could be classified according to SSC. Although SSC is very efficient, it is more accurately a descriptive terminology for clinical ictal events than a classification system. PMID- 12076101 TI - Anticonvulsant profile and mechanism of action of propranolol and its two enantiomers. AB - The anticonvulsant properties of the ss-adrenoceptor antagonist propranolol and its two enantiomers were examined in various screening tests in order to characterize the anticonvulsant profile as well as the possible molecular mechanism of action. These compounds dose-dependently raised the threshold for tonic electroshock seizures in mice and were effective in the traditional maximal electroshock test (ED (50)s 15- 20 mg kg (-1)i.p.). In combination with clinically used antiepileptics, the anticonvulsant effectiveness of the latter was significantly increased. In the pentylenetetrazol (85 mg kg (-1)s.c.) seizure threshold test, ( +/-)- and ( +)-propranolol were not effective in preventing clonic seizures. In unrestrained rats with chronically implanted electrodes in the dorsal hippocampus, propranolol and its ( +)-enantiomer equieffectively reduced the duration of electrically-evoked hippocampal afterdischarges (10 and 20 mg kg (-1)i.p.) and raised the focal stimulation threshold (20 mg kg ( 1)i.p.). In amygdala-kindled rats, both drugs ( >or= 10 mg kg (-1)i.p.) reduced the seizure severity from stage 5 (generalized clonic-tonic) to stage 3 (unilateral forelimb) seizures. Furthermore, whole-cell patch-clamp experiments showed that ( +)- as well as ( -)-propranolol ( 10(-6)to 10(-4)M) depressed the fast inward sodium current in a concentration- and use-dependent manner in cultured rat cardiomyocytes and inhibited picrotoxin-induced burst firing activity of mouse spinal cord neurones in culture. In conclusion, propranolol and its two enantiomers have anticonvulsant effects in models for generalized tonic clonic and complex partial seizures which may be accounted for by the sodium channel blocking and not by the ss-adrenoceptor blocking activity. PMID- 12076103 TI - Recurrent absence status epilepticus: clinical and EEG characteristics. AB - In order to outline the clinical and EEG characteristics of recurrent absence status epilepticus (ASE), eight cases with more than two attacks of ASE were studied. Their current ages were between 13 and 84 years, and five of the patients were women. There was a history of epilepsy in five of the patients before the first ASE episode. A varying degree of confusion was the main clinical symptom with associated mild motor signs like perioral, eyelid and generalised myoclonus, seen in one, two and four patients respectively. Two of the patients had juvenile myoclonic epilepsy. One patient had an atypical form of childhood absence epilepsy characterised by recurrent ASE attacks on awakening. There were two patients with phantom absences and late onset generalised convulsions, one patient with perioral myoclonia and absences, and finally two patients with eyelid myoclonia with absences, which are proposed syndromes. On the EEGs that revealed the diagnosis of ASE, there was a marked variability of the generalised multispike and wave discharges. The EEG findings appeared to be syndrome-related with some exceptions. IV Clonazepam lead to a dramatic improvement. Our study shows that the majority of recurrent ASE cases do not fit into the International syndrome classification. PMID- 12076102 TI - Assessment and cost comparison of sleep-deprived EEG, MRI and PET in the prediction of surgical treatment for epilepsy. AB - Our aim was to determine if less expensive interictal indices can predict which epilepsy patients may benefit from the more expensive comprehensive pre-surgical evaluation. Surgical treatment was determined based on the results of a comprehensive inpatient continuous video-EEG monitoring. This evaluation included three interictal tests, which were reviewed retrospectively-2 hour-sleep-deprived electroencephalogram (SDEEG), magnetic resonance imaging (MRI), and positron emission tomography (PET). Sixty-nine patients were evaluated with 35 patients having focal resection (33 temporal, two frontal). When two or more interictal tests were positive, 77% (27 /35) went to surgery, but when one test was positive 23% (8 /34) had surgery. When all tests were negative, only a single patient (1 /13 or 7.7%) had surgery, a frontal resection. The positive predictive value for any single interictal test was 68%, while it was higher for any combination of two positive tests (77-83%). PET was the most sensitive (0.86) single interictal test, compared to SDEEG (0.66) and MRI (0.66). The odds ratio for predicting surgical treatment for a positive PET, SDEEG, or MRI was 8.57, 4.01, and 4.01, respectively. MRI was three and PET was six times the cost of a SDEEG. The combination of SDEEG and MRI had the best cost/PPV ratio. Seventy-nine percent (11 /14) of the patients with three positive tests were seizure free following focal resection compared to 43% (9 /21) when less than three tests were positive ( P0.4). Detection of tnfalpha was significantly different between the group of horses that had infiltrative inflammatory bowel disease and those with larval cyathostominosis (P=0.028) and those that were helminth negative (P=0.014). These results indicate a possible role for tnfalpha in the pathogenesis of equine infiltrative inflammatory bowel disease. PMID- 12076112 TI - An unusual mechanism of disposal of superfluous spermatozoa in vasectomised quails. AB - As part of a series of studies designed to evaluate usual and potential functions of epithelial cells in the avian epididymis under varying circumstances, adult, sexually active male Japanese quails were vasectomised for 3, 6 and 8 weeks. The testes, epididymides and deferent ducts were studied histologically and ultrastructurally. Testicular lesions were minimal, but epididymal response was profound. Non-ciliated (type I) cells of the proximal efferent duct (PED) actively phagocytized spermatozoa and their fragments. An especially noteworthy and apparently novel observation was the proliferation of the lining epithelium in parts of the PED to form additional adluminal sheets of spermiophagic epithelium which did not lie directly on basal laminae. These sheets were composed exclusively of non-ciliated cells. Spermatozoal dissolution and macrophage activity were also important mechanisms for the disposal of superfluous and incarcerated spermatozoa in the epididymis of this bird. PMID- 12076113 TI - Development of a clinical syndrome resembling haemorrhagic septicaemia in the buffalo following intravenous inoculation of Pasteurella multocida serotype B:2 endotoxin and the role of tumour necrosis factor-alpha. AB - Clinical changes and acute phase responses, including tumour necrosis factor alpha (tnfalpha), in six buffalo calves were examined following intravenous inoculation of a bolus of endotoxin (1 microg kg(-1) bodyweight in 10 ml of phosphate-buffered saline [ pbs ]) extracted from Pasteurella multocida serotype B:2, the bacterium responsible for haemorrhagic septicaemia (hs) in Asia. Endotoxin injection caused a rapid onset of clinical signs characterised by dullness, sternal recumbency, elevated rectal temperatures, excessive salivation and dyspnoea that lasted for up to 12 hours post-inoculation (p.i.). Serum concentrations of tnfalpha rose within 1 hour p.i. to reach peak values ranging between 8 and 140 ng ml(-1) at 1-2 hours p.i. and then declined rapidly to baseline levels 3-5 hours p.i. Endotoxin injection induced other acute phase changes, including a rapid leucopenia and reductions in the serum concentrations of iron and zinc and a delayed but prolonged increase in haptoglobin from 12 hours p.i. that reached a plateau from about 60 hours p.i. Three control calves injected with 10 ml pbs showed no clinical or blood compositional changes. By reproducing key signs of hs the work confirms a pivotal role of endotoxin in the pathogenesis of hs and emphasises the exquisite sensitivity of the buffalo to P multocida endotoxin. PMID- 12076114 TI - Duration of increased serum alkaline phosphatase activity in dogs receiving different glucocorticoid doses. AB - Exposure to exogenous glucocorticoids can variably increase the serum alkaline phosphatase (alp) activity, however, the duration of this effect in dogs has not been determined. In this study, three groups of ten clinically normal adult dogs were administered different types of glucocorticoids at therapeutic doses. Group 1 received prednisone 1 mg kg day(-1)p.o. for 3 weeks; Group 2 received a single dose of methylprednisone acetate 1.1 mg kg day(-1)s.c.; Group 3 received dexamethasone 0.25 mg kg day(-1)p.o. for 1 week. In Group 1 elevations were statistically significant on days 7, 14 and 21 (P<0.01, P<0.001, P<0.001, respectively). After discontinuing therapy serum alp returned to baseline levels in 7 days. In Group 2, serum alp activity remained significantly elevated for 3 weeks after therapy (P<0.05, P<0.001, P<0.01 on days 7, 14 and 21 respectively). In Group 3, serum alp levels were significantly increased after 1 week of therapy (P<0.001) returning to basal levels 2 weeks after discontinuing glucocorticoid administration. In conclusion, duration of increased serum alp activity was variable and with the protocols assessed a 3-week period for short-acting glucocorticoids and more than 4 weeks for long-acting methylprednisone may be required to return to baseline levels in all dogs. PMID- 12076115 TI - A thermal threshold testing device for evaluation of analgesics in cats. AB - A thermal analgesiometric device was developed for unrestrained cats. Heat was provided by an electrical element potted together with a temperature sensor in thermally conductive epoxy in a 5 gm probe. This was attached to an elasticated band round the cat's thorax with an inflated bladder maintaining constant pressure between probe and skin. A safety cut-off was set at 60 degrees C. End point was a skin flick, turning, or jumping. Threshold temperatures in untreated cats were around 40 degrees C and repeatable to 4 degrees C with 5, 10 or 15 minutes between tests. Threshold temperature was stable in tests at 15 minutes intervals without false positives or negatives. Tests repeated at weekly intervals were repeatable to within 4 degrees C. Treatment with the opioid analgesic pethidine increased the threshold temperatures 10.2 (6.7) degrees C 45 minutes after treatment. The device was well tolerated for at least 24 hours and the analgesic effect of an opioid was detected. The system appears suitable for use in investigations into analgesic pharmacology in cats. PMID- 12076116 TI - Distribution and repeatability of nematode faecal egg counts in dairy goats: a farm survey and implications for worm control. AB - The aims of this study were to characterise trichostrongyle egg excretion in dairy goat flocks under various epidemiological conditions, and to assess the relative importance of animals in their first lactation and goats with high levels of milk production on the total egg output of the flock. The repeatability of egg excretion was estimated on 4 different dates in 14 dairy goat farms in France. Egg outputs were positively skewed on most farms suggesting an aggregated distribution of parasites. Within-year repeatability coefficients ranged from 0.29 to 0.82 suggesting that, on most farms, animals with high egg excretion on one occasion are likely to show high rates of excretion on other occasions. Comparisons of egg outputs from the selected subgroups showed that the high producing goats contributed prominently to egg excretion, particularly at times of peak parasitism. Results from goats in their first lactation were more equivocal. These results provide a rationale for anthelmintic strategies in dairy goats based on phenotypic characters. PMID- 12076117 TI - An experimental clot model in sheep; generation of a heterologous clot and its detection in vivo using venography and (125)I labelled fibrinogen. AB - An experimental venous clot model using the lateral saphenous vein of sheep is described. Eight experimental Suffolk crossbred sheep were used. A mixture of human fibrinogen, in some cases labelled with (125)I, bovine thrombin and homologous whole blood was placed via a catheter into a surgically isolated segment of the lateral saphenous vein. The resulting heterologous clot was imaged daily for 6 days using venography, or monitored using an external gamma ray detector. Clots were radiographically detectable for the 6 days of the study. They were totally occlusive for a mean of 4.2 days (SD 2.2) and were occlusive in the immediate 24 hour period after surgery. The fibrin component of the clot was persistent (85 per cent of the initial fibrin[ogen] present after 6 days). Radiographically the clots were seen as filling defects within partially filled vessels, or their presence was inferred from the absence of filling. A collateral blood supply was apparent immediately on vessel occlusion. No adverse effects, evidence of infection or limb oedema were seen. The model provided a reproducible blood clot within the lateral saphenous vein of the sheep. Clot imaging using venography was effective and readily achieved. It is suggested that the model is useful when a stable, intravenous deposit of heterologous (e.g. human) fibrin is required in vivo at a site suitable for venography and radionucleid monitoring. PMID- 12076118 TI - Effect of diet on markers of intestinal health in dogs. AB - The effects of two popular commercial pet foods on faecal markers of microbial metabolism were investigated. Adult dogs were fed a dry, extruded diet and a moist, canned diet in a randomly assigned crossover design. Fresh faecal samples were collected for chemical and enzyme activity assays. Relative to the canned diet, the dry food resulted in decreased faecal pH and faecal indole, sulphide and ammonia concentrations and increased total short-chain fatty acid, acetic and propionic acid concentrations. Faecal beta-glucosidase, beta-glucuronidase, beta galactosidase and nitroreductase activities were decreased in dogs fed the dry diet. These changes in microbial metabolic activity are consistent with beneficial effects of the dry diet on colonic health. PMID- 12076119 TI - Anti-enteropathogenic Escherichia coli immunoglobulin Y isolated from eggs laid by immunised Leghorn chickens. AB - IgY, the egg yolk immunoglobulin, equivalent to the IgG from mammals, has been used in veterinary practice for passive immunisation against bacterial or viral infectious diseases. Enteropathogenic Escherichia coli (EPEC) is the main etiological agent of infantile diarrhoea in Brazil and other developing countries. Our aims were to isolate immunoglobulin IgY from egg yolk laid by EPEC -immunised Leghorn chickens and to study its reactivity to the antigens from this pathogen, including some virulence factors. Leghorn chickens were immunised with a bacterial suspension intramuscularly (three hens) or intravenously (three hens) or with PBS (two hens). Eggs were collected over a period of 17 weeks. IgY isolation procedures were carried out by salt precipitation (ammonium sulphate, in solid form) followed by centrifugations and dialysis. Final preparations were submitted to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS - PAGE), enzyme-linked immunosorbent assay (ELISA) and immunoblotting. All immunised animals developed good levels of antibodies reactive to whole bacteria or lipopolysaccharide (LPS), in contrast to the control ones. Immunoblottings allowed the recognition of several antigenic fractions of bacterial antigens, some of which had a molecular weight compatible with bacterial virulence factors, confirming the efficacy of the immunisation and the adequacy of the method. PMID- 12076120 TI - Pathogenicity and protective activity in pregnant goats of a Brucella melitensis Deltaomp25 deletion mutant. AB - The Brucella melitensis mutant BM 25, which lacks the major 25 kDa outer membrane protein Omp25, has previously been found to be attenuated in the murine brucellosis model. In the present study, the capacity of the Deltaomp25 mutant to colonise and cause abortions in the caprine host was evaluated. The vaccine potential of BM 25 was also investigated in goats. Inoculation of nine pregnant goats in late gestation with the B. melitensis mutant resulted in 0/9 abortions, while the virulent parental strain, B. melitensis 16M, induced 6/6 dams to abort (P<0.001, n=6). BM 25 also colonised fewer adults (P<0.05, n=6) and kids (P<0.01, n=6) than strain 16M. The Deltaomp25 mutant was found capable of transient in vivo colonisation of non-pregnant goats for two weeks post-infection. Owing to the ability of BM 25 to colonise both non-pregnant and pregnant adults without inducing abortions, a vaccine efficacy study was performed. Vaccination of goats prior to breeding with either BM 25 or the current caprine vaccine B. melitensis strain Rev. 1 resulted in 100 per cent protection against abortion following challenge in late gestation with virulent strain 16M (P<0.05, n=7). However, unlike strain Rev. 1, BM 25 does not appear to cause abortions in late gestation based on this study with a small number of animals. The B. melitensis Deltaomp25 mutant, BM 25, may be a safe and efficacious alternative to strain Rev. 1 when dealing with goat herds of mixed age and pregnancy status. PMID- 12076121 TI - Cytosolic ratio of malate dehyrogenase/lactate dehydrogenase activity in peripheral leukocytes of race horses with training. AB - The activities of the enzymes involved in the malate-aspartate shuttle and m RNA expression of malate dehydrogenase (MDH), a crucial enzyme for the NADH shuttle that produces ATP in glucose metabolism in the peripheral leukocytes of horses, were measured to investigate the change in metabolic states with training. There were no significant differences in plasma glucose and immunoreactive insulin concentrations between race horses and riding horses, used as a comparable reference. The cytosolic and mitochondrial MDH activities in leukocytes of race horses were significantly higher than those of riding horses. High activities of MDH in leukocytes of race horses were confirmed by RT - PCR analysis on the total RNA extracted from the whole blood. The cytosolic ratio of MDH /lactate dehydrogenase (LDH) activity (M/L ratio) in leukocytes of race horses was significantly higher than in those of riding horses. Increase in the M/L ratio was considered to reflect elevation of energy metabolism in animal tissues. The M/L ratio may be a useful parameter to evaluate the difference in metabolic states between race horses and riding horses. PMID- 12076122 TI - Complete coding sequence of equine inducible nitric oxide synthase. PMID- 12076123 TI - Bacteriophage T4 development in Escherichia coli is growth rate dependent. AB - Three independent parameters (eclipse and latent periods, and rate of ripening during the rise period) are essential and sufficient to describe bacteriophage development in its bacterial host. A general model to describe the classical "one step growth" experiment [Rabinovitch et al. (1999a) J. Bacteriol.181, 1687-1683] allowed their calculations from experimental results obtained with T4 in Escherichia coli B/r under different growth conditions [Hadas et al. (1997) Microbiology143, 179-185]. It is found that all three parameters could be described by their dependence solely on the culture doubling time tau before infection. Their functional dependence on tau, derived by a best-fit analysis, was used to calculate burst size values. The latter agree well with the experimental results. The dependence of the derived parameters on growth conditions can be used to predict phage development under other experimental manipulations. PMID- 12076124 TI - Location, time, and temperature dependence of digestion in simple animal tracts. AB - In this paper, we develop plug flow reactor models that simultaneously investigate how reaction and absorption, morphological differences, and temperature influence nutrient acquisition rates in simple, tubular animal guts. We present analytical solutions to the resulting reaction-advection equations that model these processes, and we obtain formulas giving the throughflow speed that maximizes the absorption rate. The model predicts that the optimal digestion speed increases as the ratio of the rate of enzyme breakdown to the rate of absorption increases. PMID- 12076125 TI - Robustness as a measure of plausibility in models of biochemical networks. AB - Theory, experiment, and observation suggest that biochemical networks which are conserved across species are robust to variations in concentrations and kinetic parameters. Here, we exploit this expectation to propose an approach to model building and selection. We represent a model as a mapping from parameter space to behavior space, and utilize bifurcation analysis to study the robustness of each region of steady-state behavior to parameter variations. The hypothesis that potential errors in models will result in parameter sensitivities is tested by analysis of two models of the biochemical oscillator underlying the Xenopus cell cycle. Our analysis successfully identifies known weaknesses in the older model and suggests areas for further investigation in the more recent, more plausible model. It also correctly highlights why the more recent model is more plausible. PMID- 12076126 TI - Recurrent inhibitory dynamics: the role of state-dependent distributions of conduction delay times. AB - We have formulated and analysed a dynamic model for recurrent inhibition that takes into account the state dependence of the delayed feedback signal (due to the variation in threshold of fibres with their size) and the distribution of these delays (due to the distribution of fibre diameters in the feedback pathway). Using a combination of analytic and numerical tools, we have analysed the behaviour of this model. Depending on the parameter values chosen, as well as the initial preparation of the system, there may be a spectrum of post-synaptic firing dynamics ranging from stable constant values through periodic bursting (limit cycle) behaviour and chaotic firing as well as bistable behaviours. Using detailed parameter estimation for a physiologically motivated example (the CA3 basket cell-mossy fibre system in the hippocampus), we present some of these numerical behaviours. The numerical results corroborate the results of the analytic characterization of the solutions. Namely, for some parameter values the model has a single stable steady state while for the others there is a bistability in which the eventual behaviour depends on the magnitude of stimulation (the initial function). PMID- 12076127 TI - A model of the early evolution of soma-to-germline feedback. AB - The V-genes of the immunoglobulin locus in vertebrates code for a part of the heavy and light chain variable regions of antibodies and are extremely variable. Steele (1979) has developed a theory that explains the evolution of adaptive immune response by a soma-to-germline flow of cDNAs derived from somatically mutated V-genes. Here we model the early evolution of soma-to-germline feedback in a population living in a changing viral environment in terms of the dynamics of an initially rare genetic modifier that controls transfer of V-genes to germ cells' DNA. It is shown that a modifier invades the population and creates a great variety of V-genes if the environment follows stepwise temporal changes, i.e. a soma-to-germline feedback machinery evolves in a population if newly derived V-alleles still play a role in protecting the population against foreign antigens in some following generations. The distribution of the age of V-genes evolves to a bell-shaped curve the width and the maximum of which depend mainly on selection strength. Two phases of modifier evolution are distinguished. In the first phase, the dynamics are slow while the number of different V-genes is small. In the second phase, when a sufficiently large number of different V-genes is created, the modifier increases faster in frequency. Linkage of V-genes and the modifier enhances the rate of evolution. PMID- 12076128 TI - A stochastic model for patterning of the cytoplasm by the saltatory movement. AB - In various cases of importance for animal physiology and development, a specific distribution of cellular components is achieved through the active transport of these components along cytoskeletal fibres by molecular motors. The pattern generating transport is stochastic; it is commonly referred to as the saltatory movement which means frequent, random change of direction of movement of individual transported particles. Inference of the distribution of the cellular components and kinetics of transitions between different patterns from parameters of the saltatory movement is the goal of the proposed theory. The theory is presented by developing a sample model for the redistribution of lipid droplets at early stages of Drosophila development, a process well studied at the quantitative level. The saltatory movement is modelled at the fundamental level as a stochastic velocity jump process. A diffusion (in the mathematical sense) model is derived from the fundamental velocity jump description as its simple and accurate approximation. This approximation reduces the number of parameters, simplifies the methods of their measurement and clarifies the relationship between the kinetics and the resulting pattern. PMID- 12076129 TI - Tangled nature: a model of evolutionary ecology. AB - We discuss a simple model of co-evolution. In order to emphasize the effect of interaction between individuals, the entire population is subjected to the same physical environment. Species are emergent structures and extinction, origination and diversity are entirely a consequence of co-evolutionary interaction between individuals. For comparison, we consider both asexual and sexually reproducing populations. In either case, the system evolves through periods of hectic reorganization separated by periods of coherent stable coexistence. PMID- 12076130 TI - Intercellular adhesion and cancer invasion: a discrete simulation using the extended Potts model. AB - We develop a discrete model of malignant invasion using a thermodynamic argument. An extension of the Potts model is used to simulate a population of malignant cells experiencing interactions due to both homotypic and heterotypic adhesion while also secreting proteolytic enzymes and experiencing a haptotactic gradient. In this way we investigate the influence of changes in cell-cell adhesion on the invasion process. We demonstrate that the morphology of the invading front is influenced by changes in the adhesiveness parameters, and detail how the invasiveness of the tumour is related to adhesion. We show that cell-cell adhesion has less of an influence on invasion compared with cell-medium adhesion, and that increases in both proteolytic enzyme secretion rate and the coefficient of haptotaxis act in synergy to promote invasion. We extend the simulation by including proliferation, and, following experimental evidence, develop an algorithm for cell division in which the mitotic rate is explicitly related to changes in the relative magnitudes of homotypic and heterotypic adhesiveness. We show that although an increased proliferation rate usually results in an increased depth of invasion into the extracellular matrix, it does not invariably do so, and may, indeed, cause invasiveness to be reduced. PMID- 12076131 TI - Single muscle fiber contraction is dictated by inter-sarcomere dynamics. AB - This paper presents first results from a study where we developed a generic framework for analysing inter-sarcomere dynamics. Our objective is to find an accurate description of a muscle as a linear motor composed of parallel and series coupled subunits. The quality of theoretical models can be tested through their ability to predict experimental observations. With the current method we have found rigorous mathematical explanations for mechanisms such as sarcomere popping, extra tension and homogenization. These phenomena have been observed for many years in single fibers experiments, yet have never been completely understood in terms of a mechanical model. Now they can be explained on a theoretical basis. Interestingly, rather simplistic descriptions of each of the various molecular components in the sarcomere (actin-myosin cross-bridges, titin and contributions from passive elastic components) are sufficient to predict these behaviors. The complexity of a real muscle fiber is addressed through rigorous coupling of the single component models in a system of differential equations. We examine the properties of the differential equations, based on a down-stripped model, which permits the derivation of analytical solutions. They suggest that the contraction characteristics of inter-connected sarcomeres are essentially dictated by the initial distribution of the sarcomeres on the force length curve and their starting velocities. The complete model is applied to show the complexity of inter-sarcomere dynamics of activated fibers in stretch-release experiments with either external force or length control. Seemingly contradictory and unexpected observations from single fiber experiments, which have hitherto been discussed with the argument of uncontrollable biological variability, turn out to be a consistent set of possible fiber responses. They result from a convolution of multiple relatively simple rules each of them defining a certain characteristics of the single sarcomere. PMID- 12076132 TI - The sulfone linker in solid-phase synthesis: preparation of 3,5-disubstituted cyclopent-2-enones. AB - The preparation of functionalized 3,5-disubstituted cyclopent-2-enones via a solid-phase sulfone linker strategy is described. Polystyrene/divinylbenzene sulfinate 1 underwent S-alkylation followed by alpha,alpha-dialkylation with cis 1,4-dichloro-2-butene to form polymer-bound 3-phenylsulfonylcyclopentenes 8. Subsequent epoxidation of the cyclopentene moiety in 8 was accomplished by treatment of mCPBA, and the resulting oxirane ring in resin 9 was opened with various nucleophiles, i.e., Grignard and cuprate reagents, azide ion, and amines. To complete the sulfone-based linker strategy, Swern or TPAP oxidation of 10 gave a transient gamma-ketosulfone, which underwent sulfinate elimination, thus cleaving the sulfone linker. Eleven 3,5-disubstituted cyclopent-2-enones (11) were prepared with this five-step process in 18-40% overall yield from solid phase benzene sulfinate 1. PMID- 12076133 TI - Enantiospecific synthesis of the phospholipase A2 inhibitor (-)-cinatrin B. AB - The first enantiospecific synthesis of phospholipase A2 (PLA2) inhibitor (-) cinatrin B (2) from the D-arabinose derivative 9 is described. The spirolactone system was formed by an Ireland-Claisen rearrangement of the allyl ester 8 followed by hydrolysis and stereoselective iodolactonization. The stereoselectivity of the rearrangement was controlled by the asymmetry in the allylic alcohol fragment. Ester (S)-8 gave the desired rearrangement product 7 and the epimer 13 in high yield as a 73:27 ratio, respectively. The final stereocenter at C2 was introduced via a chelation-controlled addition of the Grignard reagent derived from trimethylsilylacetylene to alpha-hydroxy ketone 6. Transformation of the terminal alkyne into the methyl ester 21 followed by acetal hydrolysis and selective lactol oxidation afforded cinatrin B methyl ester (22). Base hydrolysis and acid-induced relactonization then gave (-)-cinatrin B (2). PMID- 12076134 TI - Enantioselective synthesis of 12-amino alkylidenecyclopentenone prostaglandins. AB - An enantioselective synthesis of new 12-amino alkylidenecyclopentenone prostaglandins is reported. The key step of the synthesis involved a [3.3] sigmatropic rearrangement of an asymmetric allylic cyanate to elaborate an asymmetric 5-amino-1,6-diene which was further transformed into cyclopentenone by successive ring-closing metathesis reaction catalyzed by the Grubbs reagent and one-pot oxidation. A palladium-catalyzed cross-coupling reaction on a 5-iodo-1,5 diene allowed the synthesis of prostanoids with variable Rw side chains. These new compounds exhibit high cytotoxic activities. PMID- 12076135 TI - A new route for generation of alpha-lambda3-iodanyl ketones via ester exchange of (Z)-(beta-acetoxyvinyl)-lambda3-iodanes: their nucleophilic substitutions with halides and sulfur and phosphorus nucleophiles. AB - An efficient method for generation of alpha-lambda3-iodanyl ketones from (Z)-(2 acetoxyvinyl)(phenyl)-lambda3-iodanes was developed. The method involves ester exchange of (Z)-2-acetoxyvinyl-lambda3-iodanes with methanol in the presence of triethylamine. alpha-lambda3-Iodanyl ketones react with a variety of nucleophiles such as halides, thiols, phosphines, phosphinic acids, and phosphates, under the conditions which produce alpha-functionalized carbonyl compounds probably via an S(N)2 pathway. PMID- 12076136 TI - Photocatalyzed [2 + 2 + 2]-cycloaddition of nitriles with acetylene: an effective method for the synthesis of 2-pyridines under mild conditions. AB - The photocatalyzed [2 + 2 + 2]-cycloaddition of nitriles with 2 equiv of acetylene to 2-pyridines can be carried out under mild conditions and represents a valuable extension to common synthetical methods. For the ideal wavelength range (350-500 nm), lamps as well as sunlight can be used. Working at room temperature and in organic solvents such as toluene or hexane as well as in water gives satisfying results in many cases. However, it is also possible to vary the solvent and the reaction temperature of the photocatalyzed synthesis and to choose, with respect to the specific substrate, specific requirements for this particular reaction and general requirements of the method. This simple and selective method derives its potential mainly from the large variety of applicable nitriles. Suitable substrates include (functionalized) aliphatic and aromatic nitriles as well as cyanamides derived from secondary amines. PMID- 12076137 TI - Tricalixarenes and pentacalixarenes: synthesis and complexation studies. AB - Tricalix[4]arene 4, tricalix[5]arene 14, and pentacalix[4]arene 10 have been synthesized from O-alkylcalixarene mono- and dialdehydes by a two-step conversion to the corresponding monoethynyl ketones or diethynyl ketones followed by aryne trimerization in refluxing DMF containing a dialkylamine. The tricalixarenes 4 and 14 were converted, in turn, to calixarenes 6 and 16, which carry OH groups on the lower rim and methylenes as the bridging moieties to the benzene ring. Complexation studies with the tricalix[5]arene 16 show that it forms (a) a 1:3 complex with N,N-dimethylethylenediamine in which each calixarene unit contains a molecule of the amine, (b) a 1:1 complex with tris(aminomethyl)amine in which each calixarene unit contains one of the three arms of the guest, and (c) a 1:1 complex with C60 in which the guest presumably resides in the cavity provided by the three calixarene units acting cooperatively. PMID- 12076138 TI - Synthesis of alpha-D-Galp-(1-->3)-beta-D-Galf-(1-->3)-D-man, a terminal trisaccharide of Leishmania type-2 glycoinositolphospholipids. AB - The synthesis of alpha-D-Galp-(1-->3)-beta-D-Galf-(1-->3)-D-Man, present in the type-2 glycoinositolphospholipids and in the core of the lipophosphoglycan of Leishmania, is described. The glycosyl aldonolactone approach, followed by reduction of the lactone with diisoamylborane, was utilized for the introduction of the internal galactofuranosyl unit and the trichloroacetimidate method for the O-glycosidation reaction. A high-yield synthesis of the beta-D-Galf-(1-3)-D-Man unit, also present in the lipopeptidophosphoglycan of Trypanosoma cruzi, is reported. PMID- 12076139 TI - Thermal rearrangements of spiro[2.4]hepta-1,4,6-trienes. AB - Thermolysis of spiro[2.4]hepta-1,4,6-triene (1a) at 50 degrees C yielded bicyclo[3.2.0]hepta-1,3,6-triene (5), which dimerized in two different fashions to form cyclobutanes. The 1,2-dimethyl and 1-propyl derivatives of 1a also rearranged at 50 degrees C, but at a faster rate, each yielding a pair of cyclobutane dimers. The structures of these symmetrical dimers were investigated by 1D and 2D NMR and NOE difference spectroscopy. Ab initio calculations indicated that the two strained olefins 1a and 5 had comparable energies about 50 kcal/mol lower than norborna-1(7),2,5-triene, which was thus excluded as a reaction intermediate. PMID- 12076140 TI - Carbohydrate carbocyclization by a zinc-mediated tandem reaction and ring-closing enyne metathesis. AB - Methyl 5-deoxy-5-iodo-pentofuranosides are reductively ring-opened and propargylated in a tandem fashion in the presence of zinc. The 1,7-enynes thus obtained are subjected to ring-closing enyne metathesis with catalyst B to produce functionalized 1-vinyl cyclohexenes. By adding BnNH(2) to the tandem reaction, an amino group can be introduced in the 1,7-enyne products. Addition of 2-TMS-ethynylcerium(III) chloride after the reductive ring-opening produces the corresponding 1,6-enynes. Further annulation of the product 1,3-dienes can be achieved through a Diels-Alder reaction with good control of stereochemistry. These procedures constitute efficient methods for rapid carbocyclization and annulation of carbohydrates to produce a variety of functionalized five- and six membered ring systems. PMID- 12076141 TI - Efficient and general synthesis of 5-(alkoxycarbonyl)methylene-3-oxazolines by palladium-catalyzed oxidative carbonylation of prop-2-ynylamides. AB - A variety of prop-2-ynylamides have been carbonylated under oxidative conditions to give oxazolines, oxazolines with chelating groups, and bisoxazolines bearing an (alkoxycarbonyl)methylene chain at the 5 position in good yields. The cyclization-alkoxycarbonylation process was carried out in alcoholic media at 50 70 degrees C and under 24 bar pressure of 3:1 carbon monoxide/air in the presence of catalytic amounts of 10% Pd/C or PdI2 in conjunction with KI. Cyclization occurred by anti attack of an oxygen function on the palladium-coordinated triple bond, followed by stereospecific alkoxycarbonylation, strictly resulting in E stereochemistry. The structures of representative oxazolines and bisoxazolines have been determined by X-ray diffraction analysis. PMID- 12076142 TI - Electrochemically-induced spirolactonization of alpha-(methoxyphenoxy)alkanoic acids into quinone ketals. AB - Anodic oxidation of two series of alpha-(2)- and alpha-(4-methoxyphenoxy)alkanoic acids were studied both at the analytical and preparative scales in order to delineate mechanistic aspects of electrochemically induced spirolactonization and to develop synthetically useful orthoquinone bis- and monoketals. Although alpha monomethylated carboxylic acids and acetic acid derivatives do not undergo any spiroannulation, alpha-dimethylated carboxylic acids furnished spirolactones in high yields. A gem-dimethyl effect is invoked to explain these differences in cyclization capacity. Electrooxidation conditions can be selected to furnish either quinone spirolactone bis- or monoketals. Chemoselective monohydrolysis of bisketals can also be accomplished in a stepwise fashion to furnish the corresponding spirolactone monoketals, but the ortho compound unfortunately dimerized in situ via a Diels-Alder process. An ECEC pathway is proposed to rationalize the observed spirolactonizations on the basis of cyclic voltammetry analyses. PMID- 12076143 TI - Toward a general strategy for the synthesis of 3,4-dihydroxyprolines from pentose sugars. AB - A general strategy is proposed, wherein a pentose sugar gamma-lactone can be converted, via a series of nine reactions, to a 3,4-dihydroxyproline, suitably protected for use in peptide synthesis. Thus, D-ribonolactone (6) has been converted to N-fluorenylmethoxycarbonyl-3,4-di-O-tert-butyldimethylsilyloxy-D-2,3 cis-3,4-cis-proline (7) in 18.9% overall yield. Likewise, L-arabinonolactone (11) has been converted to N-fluorenylmethoxycarbonyl-3,4-di-O-tert butyldimethylsilyloxy-L-2,3-cis-3,4-trans-proline (36) in 13.7% overall yield and L-lyxonolactone (12) to N-fluorenylmethoxycarbonyl-3,4-di-O-tert butyldimethylsilyloxy-L-2,3-trans-3,4-cis-proline (37) in 11.2% overall yield. These building blocks have also been fully deprotected to give the free amino acids. We believe that this series of reactions ought to be applicable to the synthesis of any of the eight stereoisomers of 3,4-dihydroxyproline, by judicious selection of the pentose starting material. PMID- 12076145 TI - Novel process for generating useful electrophiles from common anions using Selectfluor fluorination agent. AB - In the present work, the electrophile equivalents Cl+, Br+, SCN+, and NO2+ are generated from their respective sodium, potassium, and in some cases ammonium salts (M+X-) by reaction with Selectfluor electrophilic fluorination agent in acetonitrile solution at room temperature. These generated electrophilic species subsequently react in situ with a variety of aromatic substrates containing one or more substituent groups including H, F, Cl, CH3, COOH, C(O)CH3, NO2, and OR' and NR'R' ' where R' and R' ' are H or CH3. The resulting substitution products are, in most cases, isolable as pure compounds in high yield. Variations in the process include the use of other anions, electrophilic fluorination agents, and solvents. PMID- 12076144 TI - Synthesis of alpha- and beta-glycosyl asparagine ethylene isosteres (C-glycosyl asparagines) via sugar acetylenes and Garner aldehyde coupling. AB - A convergent approach has been developed for the synthesis of C-glycosyl amino acids in which the glycinyl moiety CH(NH2)CO2H is connected to the anomeric center of the sugar residue by a three carbon atom tether. Essentially, these compounds are isosteres of N-glycosyl asparagines in which the amide group has been replaced by an ethylene bridge. Following the coupling of alpha- or beta-D linked lithium C-glycoside acetylides with N-Boc D-serinal acetonide (Garner aldehyde), the resulting adducts were transformed into the final N-Boc-C-glycosyl alpha-aminopentanoic acids via reduction of the triple bond, deoxygenation, and oxidative cleavage of the oxazolidine ring. By this protocol, 12 C-glycosyl asparagines, six pairs of alpha- and beta-anomers, have been prepared incorporating the gluco, galacto, manno, and the corresponding 2-acetamido-2 deoxy residues. PMID- 12076146 TI - Kinetics and mechanism of the phenolysis of asymmetric diaryl carbonates. AB - The reactions 4-methylphenyl 4-nitrophenyl carbonate (MPNPC), 4-chlorophenyl 4 nitrophenyl carbonate (CIPNPC), 4-methylphenyl 2,4-dinitrophenyl carbonate (MPDNPC), and 4-chlorophenyl 2,4-dinitrophenyl carbonate (CIPDNPC) with a homogeneous series of phenoxide anions are subjected to a kinetic investigation in aqueous solution (25.0 degrees C, ionic strength 0.2 M (KCI)). Under an excess of phenoxide with respect to the substrate, all of these reactions obey pseudo first-order kinetics and are first order in phenoxide. The Bronsted-type plots for the nucleophilic rate constants (k(N)) are linear, with slopes beta = 0.48 (MPNPC), 0.67 (ClPNPC), 0.41 (MPDNPC), and 0.32 (ClPDNPC). The magnitude of these slopes and the absence of a curvature in the Bronsted plot at pK(a) = 7.1 for the CIPNPC reactions are consistent with concerted mechanisms (one step). The carbonates MPDNPC and ClPDNPC are more reactive than MPNPC and CIPNPC, respectively, toward phenoxide nucleophiles. This can be explained by the presence of a second nitro group in the nucleofuge of the dinitro derivatives, which (i) leaves their carbonyl carbon more positively charged, making them better electrophiles, and (ii) makes 2,4-dinitrophenoxide a better leaving group than 4-nitrophenoxide. The 4-chloro derivatives are more reactive than the corresponding 4-methyl derivatives. This should be due to the greater electron withdrawal of 4-chloro than 4-methyl, which makes the former carbonyl more electrophilic. Comparison of the concerted phenolysis of MPNPC with the stepwise reactions of secondary alicyclic amines with the same substrate indicates that substitution of a secondary alicyclic amine group in a zwitterionic tetrahedral intermediate by a phenoxy group greatly destabilizes the intermediate. An equation is deduced for log k(N) in terms of the basicity of the nucleophile, the nonleaving moiety, and the leaving group. This equation shows that for these reactions, the sensitivity of log k(N) to the basicity of the nonleaving moiety (beta(nlg) = -0.27) is very similar to that of the nucleofuge (beta(lg) = -0.25). PMID- 12076147 TI - Synthesis of axinohydantoins. AB - A short synthesis of the hydantoin-containing marine sponge metabolites axinohydantoins is described. A key feature of the synthesis is a putative biomimetic, intramolecular cyclization of alpha-functionalized imidazolone 5, which affords the tricyclic pyrroloazepinone framework comprising 6. In addition, the conversion of imidazolones to alpha,beta-unsaturated hydantoins is outlined and represents a new approach to these heterocyclic systems. PMID- 12076148 TI - Lactone-directed intramolecular Diels-Alder cyclization: synthesis of trans dihydroconfertifolin. AB - Trienes 1 and 3 were obtained in five steps from ethyl 4-acetoxy-3-oxobutanoate and 6-iodo-3-methyl-1,3-hexadiene. Intramolecular Diels-Alder cyclization of 1 and 3 gave tricyclic lactones 2 and 4 as the major products, respectively. The key intermediate 4 was converted in two steps to trans-dihydroconfertifolin (5). PMID- 12076149 TI - Synthesis and evaluation of potential inhibitors of chondroitin AC lyase from Flavobacterium heparinum. AB - Chondroitin AC lyase from Flavobacterium heparinum degrades chondroitin sulfate glycosaminoglycans via an elimination mechanism, resulting in disaccharides or oligosaccharides with Delta4,5-unsaturated uronic acid residues at their nonreducing end. The syntheses and testing of two potential inhibitors of this lyase are described. Methyl O-(2-acetamido-2-deoxy-beta-D-galactopyranosyl)-(1- >4)-alpha-L-threo-hex-4-enopyranoside, 1, has the trigonal geometry at C5 of the uronic acid moiety expected at the transition state, yet retains the "leaving group" sugar moiety. Surprisingly, compound 1 showed no inhibition of the enzyme. The novel 5-nitro sugar, phenyl (5S)-5-nitro-beta-D-xylopyranoside, 2, is a monosaccharide nitro analogue of the natural substrate, with C5 being a carbon acid of pK(a) 8.8. The rate of reprotonation of the anion generated at this center is sufficiently low that the anion of 2 can be used directly in initial steady-state velocity measurements without significant interference from the conjugate carbon acid. The anion of compound 2 was found to be a competitive inhibitor with a K(i) value of 5 mM, whereas the conjugate acid had a K(i) value of 35 mM. PMID- 12076150 TI - Building blocks for the solution phase synthesis of oligonucleotides: regioselective hydrolysis of 3',5'-Di-O-levulinylnucleosides using an enzymatic approach. AB - A short and convenient synthesis of 3'- and 5'-O-levulinyl-2'-deoxynucleosides has been developed from the corresponding 3',5'-di-O-levulinyl derivatives by regioselective enzymatic hydrolysis, avoiding several tedious chemical protection/deprotection steps. Thus, Candida antartica lipase B (CAL-B) was found to selectively hydrolyze the 5'-levulinate esters, furnishing 3'-O-levulinyl-2' deoxynucleosides 3 in >80% isolated yields. On the other hand, immobilized Pseudomonas cepacia lipase (PSL-C) and Candida antarctica lipase A (CAL-A) exhibit the opposite selectivity toward the hydrolysis at the 3'-position, affording 5'-O-levulinyl derivatives 4 in >70% yields. A similar hydrolysis procedure was successfully extended to the synthesis of 3'- and 5'-O-levulinyl protected 2'-O-alkylribonucleosides 7 and 8. This work demonstrates for the first time application of commercial CAL-B and PSL-C toward regioselective hydrolysis of levulinyl esters with excellent selectivity and yields. It is noteworthy that protected cytidine and adenosine base derivatives were not adequate substrates for the enzymatic hydrolysis with CAL-B, whereas PSL-C was able to accommodate protected bases during selective hydrolysis. In addition, we report an improved synthesis of dilevulinyl esters using a polymer-bound carbodiimide as a replacement for dicyclohexylcarbodiimide (DCC), thus considerably simplifying the workup for esterification reactions. PMID- 12076151 TI - Stereoselective synthesis of novel types of cyclopropyl carbocyclic nucleosides containing quaternary stereogenic centers. AB - Versatile and stereocontrolled synthetic entries to novel types of cyclopropyl carbocyclic nucleosides are described. The target products have been synthesized from suitable cyclopropane precursors obtained, in turn, from olefinic compounds derived from D-glyceraldehyde as a chiral precursor. Selective manipulation of the functional groups has allowed the preparation of enantiopure nucleosides, some of them displaying opposite chirality. All these molecules contain a quaternary stereogenic carbon at C-1 or C-3 of the cyclopropane ring and bear an amino, a hydroxymethyl, or a methyl group as an additional substituent. In one instance, thymine is directly linked to the cyclopropane. A methylene unit serves as the spacer in the other synthesized nucleosides. PMID- 12076152 TI - Synthesis of zwitterionic compounds: fully saturated pyrimidinylium and 1,3 diazepinylium derivatives via the novel rearrangement of 3-oxobutanoic acid thioanilide derivatives. AB - An unusual rearrangement following cyclization of 2-anilino-2-ethoxy-3 oxothiobutanoic acid with aliphatic 1,3- as well as 1,4-diamine leads to zwitterionic derivatives of 2-hydroxypropanoic acid. Moreover, with aromatic 1,2 diamines, fused heterocyclic systems such as pteridine, quinoxaline, and pyrido[2,3-b]pyrazine are obtained. PMID- 12076153 TI - Reaction of allylzinc reagents and zinc enolates of ketones with alpha amidoalkylphenyl sulfones. AB - Alpha-amidoalkylphenyl sulfones behave as N-acylimino equivalents in the reaction with functionalized allylzinc reagents. The addition products obtained using the zinc derivative of ethyl 2-(bromomethyl)acrylate can be readily transformed into alpha-methylene-gamma-lactams using different cyclization procedures. The allylzinc reagent obtained from 3-bromo-1-acetoxy-1-propene directly affords protected 1,2-amino alcohols with a preference for the anti stereoisomer, regardless of the structure of the alpha-amidoalkylphenyl sulfone employed. This procedure can be extended to the use of zinc enolates obtained from alpha-bromo ketones and leads to the synthesis of N-protected beta-amino ketones. PMID- 12076154 TI - The rational synthesis of chlorins via rearrangement of porphodimethenes: influence of beta-substituents on the regioselectivity and stereoselectivity of pyrroline ring formation. AB - The porphodimethene rearrangement methodology reported in this paper provides for a rational, step-by-step synthesis of chlorins from readily available pyrrole precursors. The intermediate porphodimethenes are furnished directly via the '2 + 2' MacDonald condensation, or by the less symmetry-constrained '3 + 1' condensation of a tripyrrane and bis-formyl pyrrole. The synthetic route is short and highly convergent, especially in the case of the '3 + 1' approach, and furnishes chlorins in good to moderate yields. The synthesis is highly regioselective and appears to be based on the ability of the beta-substituent to stabilize excess electron density, with an electron-neutral hydrogen or an electron-withdrawing carbonyl beta-substituent demonstrating the greatest influence on the formation of the pyrroline ring. The synthesis is highly stereoselective when epimerization of the pyrroline ring beta-carbons is possible, furnishing only the trans-reduced sterioisomer. Finally, there is substantial evidence that a fifth, axial ligand is involved in the transposition of peripheral hydrogens during the rearrangement of the pi-system from metalloporphodimethene to metallochlorin. PMID- 12076155 TI - An enaminone-directed benzannulation/macrocyclization approach to cyclophane ring systems. AB - A straightforward and modular preparative approach to 1,3,5-triaroylbenzene-based functionalized cyclophane ring systems has been developed. The key cyclophane forming macrocyclization reaction was accomplished during the course of a regioselective cross-benzannulation between bis(aryl ethynyl) ketone and enaminone reactants. Macrocyclic products with ring sizes ranging from 18- to 22 membered were successfully constructed. The composition of the tether connecting the two aryl ethynyl ketone fragments can be easily varied; consequently, this method is suitable for construction of a diverse range of structurally distinct cyclophane products. To illustrate this feature, cyclophanes possessing xylyl, alkyl, di(ethylene triamine), and di(ethylene oxy) bridging units were synthesized in isolated yields of 11-46%. Three new cyclophanes (calixarene-like macrocyles 8 and 9, as well as crownophane 18) were structurally characterized by X-ray diffractometry. PMID- 12076156 TI - The regioselective mono-deprotection of 1,3-dioxa-2,2-(di-tert-butyl)-2 silacyclohexanes with BF3*SMe2. AB - The selective mono-deprotection of di-tert-butylsilylene ethers prepared from substituted 1,3-pentanediols and 2,4-hexanediols has been achieved with BF3*SMe2. The reaction conditions are compatible with esters, allyl ethers, and TIPS ethers. The resulting di-tert-butylfluorosilyl ethers are stable to various conditions including low pH aqueous solutions and silica gel chromatography; the di-tert-butylfluorosilyl ethers are readily cleaved with HF-pyridine. Substrate stereochemistry and conformation influences the efficiency of the deprotection, while the deprotection regiochemistry is consistent with coordination of boron to the sterically more accessible oxygen prior to intramolecular delivery of fluoride. PMID- 12076158 TI - Approach toward the total synthesis of griseoviridin: formation of thioethynyl and thiovinyl ether-containing nine-membered lactones through a thioalkynylation macrolactonization-hydrostannylation sequence. AB - Synthesis of the lactone core 17 of 8-epi-griseoviridin is reported. Thioethynyl derivative 11 was easily prepared via an anionic coupling reaction between acetylenic compound 9 and sulfone 10. After desilylation of 11, saponification of the resulting hydroxy ester 12 followed by a Mitsunobu macrolactonization furnished the unusual triple-bond-containing nine-membered lactone 13 in 50% yield for the last two steps (39% after recrystallization). Stannylation under Magriotis conditions led to the pure regio- and stereocontrolled vinyltin 14 (80% yield). After a Sn/I exchange, palladium-catalyzed carbonylation delivered either the ester lactone 16 in 67% yield or the propargyl amide 17 in 65% yield. Synthesis of propargyl amide 17 of the lactone core of 8-epi-griseoviridin was achieved in 11.9% overall yield from commercial L-cystin dimethyl ester (nine steps). PMID- 12076157 TI - Synthesis of alpha-galactosyl ceramide, a potent immunostimulatory agent. AB - Alpha-galactosyl ceramide has been identified to be a potent stimulatory agent for a novel immunological process, mediated by CD1 molecules. This paper describes a short and efficient synthesis of alpha-galactosyl ceramide. Starting from commercially available 2-deoxy galactose, a suitable sphingosine derivative was obtained in nine steps and 36% overall yield, which was subsequently glycosylated to give the target molecule. This flexible route will provide various glycolipids for further exploration of this interesting biological process. PMID- 12076159 TI - Spin delocalization in 1-heteroallyl monoradicals as a measure of radical stabilization by heterovinyl substituents assessed through the EPR-spectral zero field D parameter of 1,3-cyclopentanediyl triplet diradicals. AB - The EPR-spectral zero-field splitting parameter D of the localized heterovinyl substituted 1,3-cyclopentanediyl triplet diradicals T, generated in a 2 methyltetrahydrofuran (MTHF) glass matrix at 77 K through the photochemical deazetation of the corresponding azoalkanes 1-13, is a quantitative measure of the spin-density (rho) variation by the substituents at the radical site in the 1 heteroallylic radicals. From these data, the radical-stabilizing ability of a variety of nitrogen-containing groups has been assessed, which includes imino and hydrazonyl functionalities. The radical stabilization in the heteroallylic radical fragment follows the order X = O < NMe < CH2 < CHMe << NOH approximately NOMe << NNHCHO approximately NNHC(O)NH2 < NPh approximately NNMe2 < NNH2 < CHPh < NNHPh. The lowest D values have been found for the hydrazonyl-substituted derivatives, which implies the lowest spin density at the carbon center and, thus, the most efficacious radical stabilization through spin delocalization. This superdelocalization may be rationalized in terms of nitrogen-centered radical-cationic structures. Localization of the spin at the terminal atom is resisted through the electronegativity effect (O < N < C). PMID- 12076160 TI - Dithienylethenes with a novel photochromic performance. AB - Dithienylethenes with low decoloration quantum yields and thermal reversibility at high temperature above 100 degrees C were prepared. Introduction of bulky alkoxy substituents at 2- and 2'-positions of the thiophene rings strongly suppressed the cycloreversion quantum yields. The quantum yields were lower than 10(-3), and the photogenerated color remained stable enough under room light. On the other hand, the bulky alkoxy substituent decreased the thermal stability of the colored closed-ring isomers at high temperature. The color of the dithienylethene with cyclohexyloxy substituents faded out in less than 1 min at 160 degrees C. PMID- 12076161 TI - Absolute configuration of secondary alcohols by 1H NMR: in situ complexation of alpha-methoxyphenylacetic acid esters with barium(II). AB - A novel methodology that allows the assignment of the absolute configuration of chiral secondary alcohols by NMR using only one derivative is presented. All that is needed is (a) the derivatization of the alcohol of unknown configuration with one enantiomer--either the (R)- or the (S)--of alpha-methoxyphenylacetic acid (MPA), (b) the recording of the 1H NMR spectrum of the resulting ester in MeCN d3, and (c) addition of a barium(II) salt [i.e. Ba(ClO4)2] to the NMR tube till saturation and recording of a second spectrum. The assignment of the R/S configuration to the alcohol takes a few minutes and consists on the comparison of the signs of the shifts (Deltadelta(Ba)) produced by addition of the barium(II) with those predicted for the (R) and the (S) enantiomers in accordance to a simplified model that reflects the conformational changes produced by the complexation with barium and their consequences in the chemical shifts. These conformational changes are based on experimental NMR and CD results showing that the formation of a barium(II) complex with the MPA ester moves the conformational equilibrium between syn- (sp) and anti-periplanar (ap) forms toward the most stable ones (sp), and that this leads to the increase of the shielding caused by the MPA phenyl group on a certain substituent of the alcohol. In addition, ab initio Hartree-Fock (HF) and density functional theory (DFT) calculations provide further evidence on the formation, structure, and stability of the complexes with Ba2+, Mg2+, and the influence of the solvent. The general applicability of this methodology and the reliability of the configurational assignment were assured by the study of about twenty alcohols of known configuration and diverse structural features. Its scope and limitations have also being established and other representative cations (i.e. Li+, Rb+, Cs+, Mg2+, Ca2+, Sc3+, V3+, Zn2+) were also evaluated. The procedure proposed is simple, fast, and cheap because it requires a very small amount of sample, only one derivatization, and the recording of only two 1H NMR spectra at rt. A graphical guide to facilitate the application of this new method is included at the end of the paper. PMID- 12076162 TI - Biosynthesis of terpenes. Preparation of (E)-1-hydroxy-2-methyl-but-2-enyl 4 diphosphate, an intermediate of the deoxyxylulose phosphate pathway. AB - (E)-1-hydroxy-2-methyl-but-2-enyl 4-diphosphate (E-6) was synthesized in six reaction steps from hydroxyacetone (9) and (ethoxycarbonylmethenyl) triphenylphosphorane (11) with an overall yield of 38%. The compound was shown to be identical with the product of IspG protein, which serves as an intermediate in the nonmevalonate terpene biosynthetic pathway. PMID- 12076163 TI - Phosphine-catalyzed annulation of thioamides and 2-alkynoates: a new synthesis of thiazolines. AB - The annulation of thioamides with 2-alkynoates and 2,3-dienoates under the catalysis of tri-n-butylphosphine was described. The annulation reaction provided a new entry to thiazolines, particularly those with 2-aryl substituents. PMID- 12076164 TI - A practical procedure for the large-scale preparation of methyl (2R,3S)-3-(4 methoxyphenyl)glycidate, a key intermediate for diltiazem. AB - A practical synthesis of methyl (2R,3S)-3-(4-methoxyphenyl)glycidate (-)-2, a key intermediate for diltiazem (1), was developed. Treatment of methyl (E)-4 methoxycinnamate 3 with chiral dioxirane, generated from chiral ketone 4, provided (-)-2 in 77% ee and 89% yield. The crude mixture of (-)-2 and 4 was efficiently separated by the use of novel and simple equipment performing a lipase-catalyzed transesterification and a continuous dissolution and crystallization to furnish the optically pure (-)-2 and recovery of 4 in 74% and 91% yield, respectively. PMID- 12076165 TI - A kinetic study of reactions of Et3ZnLi and di-tert-butyl ketone. AB - An organozincate of composition Et3ZnLi and di-tert-butyl ketone react in toluene to form (after hydrolysis) ethyl-di-tert-butylmethanol. The rate is proportional to approximately [Et3ZnLi](-0.5)[ketone](1) when the initial concentration of Et3ZnLi is greater than that of the ketone but proportional to [Et3ZnLi](1)[ketone](-1) when the initial concentration of ketone is greater than that of Et3ZnLi. The rate of addition of Et3ZnK to di-tert-butyl ketone is <10( 4) that of Et3ZnLi. PMID- 12076166 TI - Structure-energy relationships in curved polycyclic aromatic hydrocarbons: study of benzocorannulenes. AB - B3LYP/6-31G* calculations indicate that sequential annelation of benzene rings to the rim of corannulene gradually flattens the skeleton. Pentabenzocorannulene is predicted to exhibit nearly barrierless flipping motion at room temperature. A mixed quartic-quadratic potential successfully explains the inversion barrier and curvature (given by the pi-orbital axis vector angle) relationship. PMID- 12076167 TI - Synthesis of (Z)-4-hydroxytamoxifen and (Z)-2-[4-[1-(p-hydroxyphenyl)-2-phenyl] 1butenyl]phenoxyacetic acid. AB - The synthesis of (Z)-4-hydroxytamoxifen and (Z)-2-[4-[1-(p-hydroxyphenyl)-2 phenyl]-1-butenyl]phenoxyacetic acid was accomplished using a McMurry reaction as the key step. The perfluorotolyl derivatives of the McMurry products enabled the separation of the minor undesirable geometrical isomer. The methodology proceeds without E,Z isomerization, employs a very mild final debenzylation step compatible with a large array of functional groups, and can be applied to the generation of a variety of 4-hydroxytamoxifen analogues. PMID- 12076168 TI - Triptycene quinones in synthesis: preparation of triptycene cyclopentenedione and its reactivity as a dienophile. AB - Triptycenene quinone 1 was converted to triptycene cyclopentenedione 5 through hydroxyquinone-phenyliodonium ylide formation and thermal ring contraction of the latter. Cyclopentenedione 5 reacts as a dienophile and as a dipolarophile with dienes and nitrile oxides, affording polycyclic adducts bearing the triptycene moiety. PMID- 12076169 TI - A new protocol for a regioselective aldol condensation as an alternative convenient synthesis of beta-ketols and alpha,beta-unsaturated ketones. AB - A general and convenient synthesis of beta-ketols and alpha,beta-alkenones has been achieved by a Knoevenagel condensation of a beta-ketoacid with an aldehyde in aqueous medium. Saponification of a beta-ketoester by an aqueous KOH 10% solution gives the potassium salt of the beta-ketoacid, which is condensed in situ with an aldehyde at pH 7.8-8.0, at 60 degrees C for 5-6 h. The intermediate beta-ketocarboxylate is smoothly decarboxylated in the reaction medium, giving the beta-ketol in high yield (75-90%). Acidification of the reaction mixture at pH 1 and heating at 70 degrees C under vigorous stirring for 6 h, leads directly to the corresponding alpha,beta-unsaturated ketone in good yield (65-75%). PMID- 12076170 TI - Density functional theory investigation of the reaction of isodiiodomethane with acetylene: potential utility of isodiiodomethane for cyclopropenation reactions. AB - We present density functional theory calculations for the reactions of CH2I-I and CH2I with acetylene (HC triple bond CH) to form a cyclopropene product. CH2I-I readily reacts with HC triple bond CH to form a cyclopropene product and an I2 leaving group via a rate-determining step barrier of approximately 3.9 kcal/mol (B3LYP/Sadlej-pVTZ). Calculations indicated that the CH2I radical reacts to form an iodopropenyl radical, which can close to a cyclopropene only with difficulty. Our results indicate that CH2I-I may act as an effective carbenoid to produce cyclopropenated products from alkynes. PMID- 12076172 TI - Convenient preparation of trans-arylalkenes via palladium(II)-catalyzed isomerization of cis-arylalkenes. AB - A convenient method for the isomerization of cis-arylalkenes to their trans isomers using a palladium(II) catalyst is described. The reaction conditions are mild and general across a range of arylalkenes. The synthesis of a trans resveratrol derivative from a mixture of alkene isomers was also completed. PMID- 12076171 TI - Ginkgolides: selective acetylations, translactonization, and biological evaluation. AB - Protocols for selective acetylation of the hydroxyl groups of ginkgolide C have been developed. These acetylations have given rise to various ginkgolide C acetates and iso-ginkgolide C acetates, the latter having a rearranged skeleton resulting from translactonization. These acetyl derivatives, as well as ginkgolides A and B acetates have been investigated for their ability to bind to a cloned platelet-activating factor (PAF) receptor. PMID- 12076173 TI - Total synthesis of (-)- and (+)-lentiginosine. AB - Total synthesis of (-)-lentiginosine was achieved from D-mannitol using highly stereoselective reactions. Similarly, (+)-lentiginosine was synthesized from L tartaric acid. PMID- 12076175 TI - Exercise-induced hypoalgesia and intensity of exercise. AB - Pain sensitivity has been found to be altered following exercise. A number of investigators have found diminished sensitivity to pain (hypoalgesia) during and following exercise. However, currently it is unknown whether there is a specific intensity of exercise that is required to produce this hypoalgesia response. Aerobic exercise, such as cycling and running, have been studied most often, and a number of different exercise protocols have been used in this research including: (i) increasing exercise intensity by progressively increasing the workloads; (ii) prescribing a particular exercise intensity based on a percentage of maximum; and (iii) having participants self-select the exercise intensity. Results indicate that hypoalgesia occurred consistently following high-intensity exercise. In the studies in which exercise intensity was increased by increasing workloads, hypoalgesia was found most consistently with a workload of 200 W and above. Hypoalgesia was also found following exercise prescribed at a percentage of maximal oxygen uptake (e.g. 60 to 75%). Results are less consistent for studies that prescribed exercise based on percentage of heart rate maximum, as well as for studies that let participants self-select the exercise intensity. However, there has not been a systematic manipulation of exercise intensity in most of the studies conducted in this area. In addition, the interaction between exercise intensity and exercise duration, more than likely influences whether hypoalgesia occurs following exercise. There is a need for research to be conducted in which both intensity and duration of exercise are manipulated in a systematic manner to determine the 'optimal dose' of exercise that is required to produce hypoalgesia. In addition, there is a need for more research with other modes of exercise (e.g. resistance exercise, isometric exercise) to determine the optimal dose of exercise required to produce hypoalgesia. PMID- 12076176 TI - Training techniques to improve endurance exercise performances. AB - In previously untrained individuals, endurance training improves peak oxygen uptake (VO2peak), increases capillary density of working muscle, raises blood volume and decreases heart rate during exercise at the same absolute intensity. In contrast, sprint training has a greater effect on muscle glyco(geno)lytic capacity than on muscle mitochondrial content. Sprint training invariably raises the activity of one or more of the muscle glyco(geno)lytic or related enzymes and enhances sarcolemmal lactate transport capacity. Some groups have also reported that sprint training transforms muscle fibre types, but these data are conflicting and not supported by any consistent alteration in sarcoplasmic reticulum Ca2+ ATPase activity or muscle physicochemical H+ buffering capacity. While the adaptations to training have been studied extensively in previously sedentary individuals, far less is known about the responses to high-intensity interval training (HIT) in already highly trained athletes. Only one group has systematically studied the reported benefits of HIT before competition. They found that >or=6 HIT sessions, was sufficient to maximally increase peak work rate (W(peak)) values and simulated 40 km time-trial (TT(40)) speeds of competitive cyclists by 4 to 5% and 3.0 to 3.5%, respectively. Maximum 3.0 to 3.5% improvements in TT(40) cycle rides at 75 to 80% of W(peak) after HIT consisting of 4- to 5-minute rides at 80 to 85% of W(peak) supported the idea that athletes should train for competition at exercise intensities specific to their event. The optimum reduction or 'taper' in intense training to recover from exhaustive exercise before a competition is poorly understood. Most studies have shown that 20 to 80% single-step reductions in training volume over 1 to 4 weeks have little effect on exercise performance, and that it is more important to maintain training intensity than training volume. Progressive 30 to 75% reductions in pool training volume over 2 to 4 weeks have been shown to improve swimming performances by 2 to 3%. Equally rapid exponential tapers improved 5 km running times by up to 6%. We found that a 50% single-step reduction in HIT at 70% of W(peak) produced peak approximately 6% improvements in simulated 100 km time-trial performances after 2 weeks. It is possible that the optimum taper depends on the intensity of the athletes' preceding training and their need to recover from exhaustive exercise to compete. How the optimum duration of a taper is influenced by preceding training intensity and percentage reduction in training volume warrants investigation. PMID- 12076178 TI - Mountain biking injuries: an update. AB - This article reviews the available literature regarding injuries in off-road bicyclists. Recent progress in injury research has allowed the description of several patterns of injury in this sport. Mountain biking remains popular, particularly among young males, although sales and participation figures have decreased in the last several years. Competition in downhill racing has increased, while cross-country racing has decreased somewhat in popularity. Recreational riders comprise the largest segment of participants, but little is known about the demographics and injury epidemiology of noncompetitive mountain cyclists. Most mountain bikers participating in surveys reported a history of previous injuries, but prospective studies conducted at mountain bike races have found injury rates of <1%. The most common mechanism of injury involves a forward fall over the handlebars, usually while riding downhill, which can result in direct trauma to the head, torso and upper extremities. A variety of factors can be associated with this type of fall, including trail surface irregularities, mechanical failures and loss of control. In mountain bike racing the risk of injury may be higher for women than men. Minor injuries such as abrasions and contusions occur frequently, but are usually of little consequence. Fractures usually involve the torso or upper extremities, and shoulder injuries are common. Head and face injuries are not always prevented by current helmet designs. Fatal injuries are rare but have been reported. Improvements in safety equipment, rider training and racecourse design are suggested injury prevention measures. The authors encourage continued research in this sport. PMID- 12076177 TI - Fatigue during high-intensity intermittent exercise: application to bodybuilding. AB - Resistance exercise is an activity performed by individuals interested in competition, those who wish to improve muscle mass and strength for other sports, and for individuals interested in improving their strength and physical appearance. In this review we present information suggesting that phosphocreatine depletion, intramuscular acidosis and carbohydrate depletion are all potential causes of the fatigue during resistance exercise. In addition, recommendations are provided for nutritional interventions, which might delay muscle fatigue during this type of activity. PMID- 12076179 TI - Narrow versus broad spectrum antibacterials: factors in the selection of pneumococcal resistance to beta-lactams. AB - Streptococus pneumoniae represents an interesting model to discuss the relative impact of broad versus narrow spectrum antibacterials as potential selectors for resistance. Indeed, this pathogen is responsible for potentially severe infections in the community, and has a great capacity for acquisition of resistance to antibacterial agents. It has been the focus of many studies to elucidate some unique aspects of molecular biology, including the adaptive mechanisms responsible for emergence and spread of multiresistance. In the past, the use of narrow spectrum agents was recommended in order to try to reduce the risk of selection of resistance. This concept is nowadays somewhat obsolete for several reasons. S. pneumoniae is able to acquire resistance to antibacterials belonging to different families of drugs through different molecular mechanisms. Thus, selection of multiresistant pneumococci can result from exposure to very different agents, including narrow spectrum as well as broad spectrum agents. In vitro studies have shown a different potential for selection of resistance among the beta-lactam agents. Furthermore, several studies have more or less directly established a close relationship between the level of antibacterial use and the rate of selection of resistance. In addition to the overall amount of antibacterials prescribed in the community, several other factors have been shown to influence the rate of selection of resistance, including the use of doses that are too low, the length of therapy and the duration of bacterial exposure to long acting agents compared to drugs with short half-lives. Therefore, there are three main ways to control selection and spread of resistant strains: by (i) reducing the amount of antibacterials used; (ii) using optimal dosages (avoiding underdosing) and treatments of short duration; and (iii) reducing the risk of transmission among young children attending daycare centres or kindergartens. In order to help physicians reduce the number of unnecessary prescriptions, it is important to develop rapid tests to recognise the bacterial origin of a febrile illness and even more important to detect resistance to antibacterials. However, apart from rapid diagnostic tests for streptococcal pharyngitis, those tests are not currently available. As a consequence, currently, the debate around narrow versus broad spectrum antibacterials remains a false debate. Physicians should use broad spectrum agents in many instances of upper or lower respiratory tract infection, taking into consideration the probable pathogens and the risk of (multi)resistance to antibacterials. Once rapid diagnostic are available in community practice, allowing a precise diagnosis of the offending agent and its susceptibility profile, physicians will be able to add to their current criteria the selective potential for resistance of the antibacterials that appear to be active in vitro. PMID- 12076181 TI - Combination therapy for malaria: the way forward? AB - Unless new strategies are deployed to combat malaria, the already enormous health and economic burden related to the disease in tropical countries is bound to worsen. The main obstacle to malaria control is the emergence of drug resistant strains of Plasmodium falciparum. As for HIV/AIDS and tuberculosis, the use of combinations of antimalarial drugs reduces the risk of selecting for resistant mutants of the plasmodial parasites. In large field trials, the combination of an artemisinin derivative and a partner drug with an unrelated mode of action (in this case mefloquine), has shown a remarkable double effect: preventing the emergence and spread of drug resistance, and interrupting the transmission of P. falciparum. This has opened the way for a new approach to the deployment of antimalarial drugs. Coupled with early detection and confirmed diagnosis, this strategy represents the only way forward in the chemotherapy of malaria. Massive economic assistance will be needed to detect and treat adequately the estimated 500 million cases of malaria per year, but without radical action there is no prospect of 'Rolling Back' malaria. PMID- 12076183 TI - Olmesartan medoxomil. AB - Olmesartan medoxomil is a nonpeptide angiotensin II receptor antagonist which selectively and competitively inhibits the type 1 angiotensin II receptor without affecting other receptors regulating the cardiovascular system. In well designed randomised trials, olmesartan medoxomil was significantly more effective than placebo, and at dosages of 10 to 20 mg/day was at least as effective as atenolol 50 to 100 mg/day in reducing diastolic blood pressure (DBP). At dosages of 5 to 20 mg/day, olmesartan medoxomil was more effective than captopril 12.5 to 50mg twice daily at lowering seated DBP in patients with mild to moderate hypertension in a dose titration study. Reductions in seated DBP were greater with olmesartan medoxomil 10 to 20 mg/day than losartan 50 to 100 mg/day. Olmesartan medoxomil at 20 mg/day was more effective in lowering seated DBP than losartan 50 mg/day, valsartan 80 mg/day or irbesartan 150 mg/day, and was more efficacious than losartan 50 mg/day or valsartan 80 mg/day at reducing 24-hour ambulatory systolic blood pressure. Olmesartan medoxomil has shown no clinically important pharmacokinetic interactions with digoxin, warfarin or antacid (aluminium magnesium hydroxide). Adverse events were infrequent in clinical studies of olmesartan medoxomil and were similar to those attributed to placebo. With olmesartan medoxomil, the frequency of dizziness was higher than with placebo but similar to that occurring with losartan, valsartan and irbesartan. PMID- 12076182 TI - Treatment of heroin (diamorphine) addiction: current approaches and future prospects. AB - New pharmacological treatments for heroin (diamorphine) addiction include drugs that reduce opiate withdrawal symptoms and agents that are given during the maintenance phase of treatment. A variety of different types of pharmacological agents (opioid agonists, partial opioid agonists, opioid antagonists and alpha(2) adrenoreceptor agonists) are reviewed and the evidence of their use during managed withdrawal and maintenance are presented. Experimental approaches attempting to reduce the time of opiate withdrawal and to accelerate the transition to abstinence are being developed. The combination tablet of buprenorphine and naloxone that is to be introduced for office-based maintenance is currently undergoing intense evaluation in the US. This new approach may facilitate the expansion of treatment while reducing the potential for medication diversion and intravenous use. PMID- 12076188 TI - Gliclazide modified release. AB - Gliclazide modified release (MR) is a new formulation of the drug gliclazide and is given once daily. The hydrophilic matrix of hypromellose-based polymer in the new formulation effects a progressive release of the drug which parallels the 24 hour glycaemic profile in untreated patients with type 2 diabetes mellitus. The formulation shows high bioavailability and its absorption profile is unaffected by coadministration with food. Mean plasma glucose levels are significantly reduced over a 24-hour period in patients with type 2 diabetes mellitus treated with gliclazide MR once daily, in both fasting and postprandial states. No cardiovascular ATP-sensitive potassium channel interaction has been observed at therapeutic concentrations of gliclazide MR. Gliclazide MR has also demonstrated antioxidant properties that are independent of glycaemic control. In a randomised, double-blind, multicentre study, gliclazide MR 30 to 120 mg once daily showed similar efficacy to gliclazide immediate release (IR) 80 to 320 mg/day (in divided doses for doses >80 mg) in patients with type 2 diabetes mellitus over a 10-month period, reducing glycosylated haemoglobin (HbA(1c)) and fasting plasma glucose (FPG) to a similar extent. The drug appeared most efficacious in patients who had previously been treated by diet alone, where significant reductions in HbA(1c) from baseline of 0.9% and 0.95% were seen at 10 and 24 months. Similarly, a sustained effect of gliclazide MR was observed in a subgroup of elderly patients defined a priori; HbA(1c) was decreased to a similar degree to that observed in the general study population. Gliclazide MR showed similar tolerability to gliclazide IR after 10 months' treatment in the randomised trial. The most commonly observed adverse events were arthralgia, arthritis, back pain and bronchitis (each <5%). Bodyweight remained stable. In this study no episodes of nocturnal hypoglycaemia or hypoglycaemia requiring third party assistance were observed during treatment with gliclazide MR. Episodes of symptomatic hypoglycaemia were infrequent, occurring in approximately 5% of patients. PMID- 12076180 TI - The role of the angiotensin system in cardiac glucose homeostasis: therapeutic implications. AB - Resistance to the metabolic actions of insulin is thought to play a determining role in the aetiology of a great variety of disorders, including essential hypertension, accelerated atherosclerosis and cardiomyopathies. ACE inhibitors are recognised as being highly effective therapy for hypertension and cardiac insufficiency, and have a more beneficial effect on survival rate than expected on the basis of known mechanisms of action. The mechanism responsible for these extremely positive effects are just beginning to be understood and appear to be linked to the effects these drugs have on metabolism. The relationship between the insulin and angiotensin II (Ang II) signalling pathways needs to be fully clarified in order to prevent or correct the target organ damage resulting from changes in the cross-talk of these two hormonal systems. In recent years, Ang II has been shown to play a central role in cardiovascular and neuroendocrine physiology as well as in cellular cycle control. Moreover, the fact that Ang II utilises the insulin-receptor substrate (IRS)-1 to relay signals towards their intracellular destination, provides the biochemical explanation of how these two systems interact in a healthy organism and in a diseased one. Since it is overactivity of the renin-angiotensin system that seems to impair the intracellular response to insulin signalling, cardiovascular drugs that modulate the cellular transmission of Ang II have attracted particular interest. As well as the already widely-used ACE inhibitors, selective blockers of the Ang II type 1 receptor (AT(1)) have been shown to be clinically effective in the control of haemodynamic parameters, but with perhaps a less striking effect on glucose homeostasis. Many trials have investigated the effect of Ang II blockade on systemic glucose homeostasis. The inhibition of Ang II by ACE-inhibitors frequently showed a positive effect on glycaemia and insulin sensitivity, while information on the effects of AT(1) receptor antagonists on glucose homeostasis is more limited and controversial. An important limitation of these studies has been the short treatment and follow-up periods, even for the 'so called' long term studies which were only 6 months. Several investigators have focused on the effects of the nuclear factors involved in gene transcriptions, especially with respect to the agonists/antagonists of peroxisome proliferator-activated receptors (PPARs) and their intriguing interconnections with the insulin and Ang II subcellular pathways. In fact, in vitro and in vivo experimental studies have shown that thiazolidinediones (selective PPAR-gamma ligands) are not only powerful insulin sensitisers, but also have anti-hypertensive and anti atherosclerotic properties. In addition to conventional pharmacological approaches, attempts have been made to use genetic transfer in the treatment of cardiovascular and metabolic disorders. The development of powerful viral vectors carrying target genes has allowed us to restore the expression/function of specific proteins involved in the cellular mechanism of insulin resistance, and research now needs to move beyond animal models. Although a clearer picture is now emerging of the pathophysiological interaction between insulin and Ang II, especially from pre-clinical studies, there is much to be done before experimental findings can be used in daily clinical practice. PMID- 12076191 TI - Perindopril: in congestive heart failure. AB - Perindopril is a long-acting ACE inhibitor, acting through its only active metabolite perindoprilat. It inhibits the renin-angiotensin system by preventing both the conversion of angiotensin I to angiotensin II and the degradation of bradykinin, thereby reducing the vasoconstriction and left ventricular remodelling characteristic of heart failure. Perindopril 4mg significantly improved a range of haemodynamic parameters in single-dose and long-term (8 weeks and 3 months) studies involving patients with congestive heart failure (CHF), with little or no effect on blood pressure or heart rate. In randomised, double blind, placebo-controlled clinical trials conducted over 3 months and a large noncomparative study (up to 30 months), perindopril 4mg once daily significantly increased exercise tolerance and reduced symptoms of heart failure in patients with mild to moderate CHF. Perindopril 4mg once daily is generally well tolerated in patients with mild to moderate CHF. In a large noncomparative study the most commonly reported adverse clinical event was cough, which led to 2.8% of patients discontinuing treatment. In short-term comparative trials there was a significantly lower incidence of first-dose hypotension following the recommended starting dose of perindopril 2mg than after the equivalent starting doses of captopril, enalapril and lisinopril. PMID- 12076197 TI - A piece of my mind. Dead man talking. PMID- 12076194 TI - Ramipril: a review of its use in the prevention of cardiovascular outcomes. AB - Ramipril, an angiotensin-converting enzyme (ACE) inhibitor, is a prodrug which is rapidly hydrolysed after absorption to the active metabolite ramiprilat. Earlier trials have shown that ACE inhibitors, when given to patients with low ejection fractions, have reduced the relative risk of myocardial infarction (MI) and other ischaemic events by 14 to 23%. Subsequently, the double-blind, randomised, placebo-controlled, multicentre Heart Outcomes Prevention Evaluation (HOPE) study has shown that, in patients who are not known to have low ejection fraction or heart failure but are at increased risk for developing cardiovascular events, ramipril reduced the incidence of stroke, MI and death due to cardiovascular disease. Results from the HOPE study, in which 9297 patients were randomised to receive either ramipril 10 mg/day or placebo for a mean of 4.5 years, indicate that ramipril reduced the relative risk of the composite outcome of MI, stroke and cardiovascular death by 22%. The incidence of the composite outcome was significantly lower in the ramipril group than in the placebo group (14.0 vs 17.8%). Patients who received ramipril, compared with placebo recipients, had a significantly decreased incidence of stroke, MI or death due to cardiovascular disease (3.4 vs 4.9%, 9.9 vs 12.3% and 6.1 vs 8.1%, respectively). The relative risk of death from any cause was reduced among patients who received ramipril. In addition, treatment with ramipril reduced as the incidence of revascularisation procedures, and, among patients with diabetes mellitus, ramipril reduced the incidence of complications related to diabetes mellitus, including the development of overt nephropathy. Moreover, in patients without a previous diagnosis of diabetes mellitus, ramipril, compared with placebo, significantly reduced the development of diabetes mellitus. Furthermore, compared with patients receiving placebo, patients receiving ramipril had a reduced rate of progression of carotid artery wall thickness. CONCLUSION: Ramipril 10 mg/day can significantly reduce the incidence of MI, stroke or death from cardiovascular causes in patients aged > or =55 years who are at increased risk for the development of ischaemic cardiovascular events due to a history of stroke, coronary artery disease (with controlled blood pressure), diabetes mellitus plus at least one other risk factor or peripheral vascular disease but no heart failure or low ejection fraction. Therefore, in addition to dietary and lifestyle modifications, ramipril should be an integral part of secondary prevention therapy in patients at increased risk for the development of cardiovascular events. PMID- 12076198 TI - Contemporary concepts of the pathogenesis and management of preeclampsia. PMID- 12076199 TI - Economic development key to healthier world. PMID- 12076195 TI - Enoxaparin: an update of its clinical use in the management of acute coronary syndromes. AB - Enoxaparin (enoxaparin sodium) is a low molecular weight heparin (LMWH) indicated for use in the treatment of ischaemic complications of unstable angina and non-Q wave myocardial infarction (MI). Unfractionated heparin (UFH) has for many years represented the standard in anticoagulant therapy for patients with acute coronary syndromes; however, recent studies suggest that enoxaparin is also a viable option for anticoagulant therapy in these patients. The ESSENCE (Efficacy and Safety of Subcutaneous Enoxaparin in Non-Q wave Coronary Events) and the TIMI 11B (Thrombolysis in Myocardial Infarction) studies reported that twice daily enoxaparin was significantly more effective than a continuous infusion of UFH in reducing the composite triple endpoint of death, MI, or recurrent angina or urgent revascularisation. Follow-up of both patient populations showed continued benefit associated with enoxaparin. Enoxaparin has been compared with tinzaparin in the treatment of unstable coronary artery disease using a nonblind study design. There was no difference between treatment groups in the therapeutic endpoints. Three nonblind studies have also compared the effects of enoxaparin and UFH in patients receiving thrombolytic therapy following acute MI. The HART II (Heparin and Aspirin Reperfusion Therapy), the ASSENT 3 (Assessment of the Safety and Efficacy of a New Thrombolytic Regimen) and the ENTIRE-TIMI 23 (Enoxaparin and Tenecteplase with or without glycoprotein IIb/IIIa Inhibitor as Reperfusion strategy in ST Elevation MI - Thrombolysis in Myocardial Infarction) studies have revealed that enoxaparin in combination with alteplase or tenecteplase is at least equivalent (HART II and ENTIRE-TIMI 23), and possibly superior (ASSENT 3) to UFH. Enoxaparin is administered as a twice-daily subcutaneous injection. In contrast, UFH is administered as an intravenous infusion which requires routine monitoring of the activated partial thromboplastin time to ensure adequate levels of anticoagulation are maintained. During the acute phase of the the ESSENCE and TIMI 11B studies, the incidence of major bleeding was similar in patients receiving enoxaparin to that in patients receiving UFH. In contrast, the rates of minor bleeding were higher in patients receiving enoxaparin than in those receiving UFH throughout these studies. CONCLUSIONS: Data from the ESSENCE, TIMI 11B and ASSENT 3 studies have prompted calls for those LMWHs which have been shown to be superior to UFH, to be considered as first choice treatment for anticoagulation in unstable coronary syndromes. To date, these suggestions are not reflected in current guidelines which consider UFH and LMWHs equally. Irrespective, the clinical data reported in this review support the use of enoxaparin in the treatment of acute coronary syndromes. These data suggest that enoxaparin shows certain clinical and practical advantages over standard treatment with UFH and represents an important development in the treatment of acute coronary syndromes. PMID- 12076205 TI - Should consumers trust hospital quality report cards? PMID- 12076200 TI - Harvard Macy Institute helps physicians become better educators and change agents. PMID- 12076206 TI - Should consumers trust hospital quality report cards? PMID- 12076207 TI - Should consumers trust hospital quality report cards? PMID- 12076209 TI - Osteopontin as a biomarker for ovarian cancer. PMID- 12076210 TI - Osteopontin as a biomarker for ovarian cancer. PMID- 12076211 TI - Osteopontin as a biomarker for ovarian cancer. PMID- 12076213 TI - Depression in an elderly man. PMID- 12076214 TI - Depression in an elderly man. PMID- 12076216 TI - Use of feeding tubes in nursing home residents with severe cognitive impairment. PMID- 12076217 TI - Fluvastatin for prevention of cardiac events following successful first percutaneous coronary intervention: a randomized controlled trial. AB - CONTEXT: Percutaneous coronary intervention (PCI) is associated with excellent short-term improvements in ischemic symptoms, yet only three fifths of PCI patients at 5 years and one third of patients at 10 years remain free of major adverse cardiac events (MACE). OBJECTIVE: To determine whether treatment with fluvastatin reduces MACE in patients who have undergone PCI. DESIGN AND SETTING: Randomized, double-blind, placebo-controlled trial conducted at 77 referral centers in Europe, Canada, and Brazil. PATIENTS: A total of 1677 patients (aged 18-80 years) recruited between April 1996 and October 1998 with stable or unstable angina or silent ischemia following successful completion of their first PCI who had baseline total cholesterol levels between 135 and 270 mg/dL (3.5-7.0 mmol/L), with fasting triglyceride levels of less than 400 mg/dL (4.5 mmol/L). INTERVENTIONS: Patients were randomly assigned to receive treatment with fluvastatin, 80 mg/d (n = 844), or matching placebo (n = 833) at hospital discharge for 3 to 4 years. MAIN OUTCOME MEASURE: Survival time free of MACE, defined as cardiac death, nonfatal myocardial infarction, or reintervention procedure, compared between the treatment and placebo groups. RESULTS: Median time between PCI and first dose of study medication was 2.0 days, and median follow-up was 3.9 years. MACE-free survival time was significantly longer in the fluvastatin group (P =.01). One hundred eighty-one (21.4%) of 844 patients in the fluvastatin group and 222 (26.7%) of 833 patients in the placebo group had at least 1 MACE (relative risk [RR], 0.78; 95% confidence interval [CI], 0.64-0.95; P =.01). This result was independent of baseline total cholesterol levels (above [RR, 0.76; 95% CI, 0.56-1.04] vs below [RR, 0.77; 95% CI, 0.57-1.02] the median). In subgroup analysis, the risk of MACE was reduced in patients with diabetes (n = 202; RR, 0.53; 95% CI, 0.29-0.97; P =.04) and in those with multivessel disease (n = 614; RR, 0.66; 95% CI, 0.48-0.91; P =.01) who received fluvastatin compared with those who received placebo. There were no instances of creatine phosphokinase elevations 10 or more times the upper limit of normal or rhabdomyolysis in the fluvastatin group. CONCLUSION: Fluvastatin treatment in patients with average cholesterol levels undergoing their first successful PCI significantly reduces the risk of major adverse cardiac events. PMID- 12076218 TI - Dietary intake of antioxidants and risk of Alzheimer disease. AB - CONTEXT: Laboratory findings have suggested that oxidative stress may contribute to the pathogenesis of Alzheimer disease. Therefore, the risk of Alzheimer disease might be reduced by intake of antioxidants that counteract the detrimental effects of oxidative stress. OBJECTIVE: To determine whether dietary intake of antioxidants is related to risk of Alzheimer disease. DESIGN AND SETTING: The Rotterdam Study, a population-based, prospective cohort study conducted in the Netherlands. PARTICIPANTS: A total of 5395 participants who, at baseline (1990-1993), were aged at least 55 years, free of dementia, and noninstitutionalized and had reliable dietary assessment. Participants were reexamined in 1993-1994 and 1997-1999 and were continuously monitored for incident dementia. MAIN OUTCOME MEASURES: Incidence of Alzheimer disease, based on Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition (DSM-III-R) criteria and National Institute of Neurological and Communicative Disorders and Stroke and Alzheimer Disease and Related Disorders Association (NINCDS-ADRDA) criteria, associated with dietary intake of beta carotene, flavonoids, vitamin C, and vitamin E. RESULTS: After a mean follow-up of 6 years, 197 participants developed dementia, of whom 146 had Alzheimer disease. When adjustments were made for age, sex, baseline Mini-Mental State Examination score, alcohol intake, education, smoking habits, pack-years of smoking, body mass index, total energy intake, presence of carotid plaques, and use of antioxidative supplements, high intake of vitamin C and vitamin E was associated with lower risk of Alzheimer disease (rate ratios [RRs] per 1-SD increase in intake were 0.82 [95% confidence interval [CI], 0.68-0.99] and 0.82 [95% CI, 0.66-1.00], respectively). Among current smokers, this relationship was most pronounced (RRs, 0.65 [95% CI, 0.37-1.14] and 0.58 [95% CI, 0.30-1.12], respectively) and also was present for intake of beta carotene (RR, 0.49 [95% CI, 0.27-0.92]) and flavonoids (RR, 0.54 [95% CI, 0.31-0.96]). The associations did not vary by education or apolipoprotein E genotype. CONCLUSION: High dietary intake of vitamin C and vitamin E may lower the risk of Alzheimer disease. PMID- 12076219 TI - Dietary intake of antioxidant nutrients and the risk of incident Alzheimer disease in a biracial community study. AB - CONTEXT: Oxidative processes have been suggested as elements in the development of Alzheimer disease (AD), but whether dietary intake of vitamin E and other antioxidant nutrients prevents its development is unknown. OBJECTIVE: To examine whether intake of antioxidant nutrients, vitamin E, vitamin C, and beta carotene is associated with incident AD. DESIGN, SETTING, AND PARTICIPANTS: Prospective study, conducted from 1993 to 2000, of individuals selected in a stratified random sample of community-dwelling residents. The 815 residents 65 years and older were free of AD at baseline and were followed up for a mean of 3.9 years. They completed food frequency questionnaires an average of 1.7 years after baseline. MAIN OUTCOME MEASURE: Incident AD diagnosed in clinical evaluations with standardized criteria. RESULTS: Increasing vitamin E intake from foods was associated with decreased risk of developing AD after adjustment for age, education, sex, race, APOE epsilon 4, and length of follow-up. Relative risks (95% confidence intervals [CIs]) from lowest to highest quintiles of intake were 1.00, 0.71 (0.24-2.07), 0.62 (0.26-1.45), 0.71 (0.27-1.88), and 0.30 (0.10-0.92) (P for trend =.05). The protective association of vitamin E was observed only among persons who were APOE epsilon 4 negative. Adjustment for other dietary factors reduced the protective association. After adjustment for baseline memory score, the risk was 0.36 (95% CI, 0.11-1.17). Intake of vitamin C, beta carotene, and vitamin E from supplements was not significantly associated with risk of AD. CONCLUSION: This study suggests that vitamin E from food, but not other antioxidants, may be associated with a reduced risk of AD. Unexpectedly, this association was observed only among individuals without the APOE epsilon 4 allele. PMID- 12076221 TI - Content and design attributes of antivaccination web sites. AB - CONTEXT: Individuals searching the Internet for vaccine information may find antivaccination Web sites. Few published studies have systematically evaluated these sites. OBJECTIVES: To examine antivaccination Web site attributes and to delineate the specific claims and concerns expressed by antivaccination groups. DESIGN AND SETTING: In late 2000, using a metasearch program that incorporates 10 other search engines, we reviewed and analyzed 772 links to find 12 Web sites that promulgated antivaccination information. Analyzing links from these 12 sites yielded another 10 sites, producing a total of 22 sites for study. Using a standardized form, 2 authors (R.M.W., L.K.S.) systematically evaluated these sites based on specific content and design attributes. MAIN OUTCOME MEASURES: Presence or absence of 11 Web site content attributes (antivaccination claims) and 10 Web site design attributes. RESULTS: The most commonly found content claims were that vaccines cause idiopathic illness (100% of sites), vaccines erode immunity (95%), adverse vaccine reactions are underreported (95%), and vaccination policy is motivated by profit (91%). The most common design attributes were the presence of links to other antivaccination sites (100%of sites), information for legally avoiding immunizations (64%), and the use of emotionally charged stories of children who had allegedly been killed or harmed by vaccines (55%). CONCLUSION: Antivaccination Web sites express a range of concerns related to vaccine safety and varying levels of distrust in medicine. The sites rely heavily on emotional appeal to convey their message. PMID- 12076220 TI - Noninvasive positive-pressure ventilation for postextubation respiratory distress: a randomized controlled trial. AB - CONTEXT: Noninvasive positive-pressure ventilation (NPPV) has been demonstrated to be effective in preventing the need for endotracheal intubation in some patients who present with acute respiratory failure. It is also used for patients who develop acute respiratory distress after extubation, but there are no randomized controlled trials that address its effectiveness in this population. OBJECTIVE: To determine the effectiveness of NPPV compared with standard medical therapy in preventing the need for endotracheal reintubation in high-risk patients who develop respiratory distress during the first 48 hours after extubation. DESIGN: Randomized, controlled, unblinded study with concealed allocation conducted between August 1, 1996 and October 31, 1999. SETTING: An intensive care unit (ICU) in an academic, tertiary care hospital in Ontario. PATIENTS: Eighty-one patients with a history of cardiac or respiratory disease or who initially required ventilatory support for more than 2 days and who developed respiratory distress within 48 hours of extubation. INTERVENTIONS: Patients were randomly assigned to receive standard medical therapy alone (supplemental oxygen to maintain oxygen saturation by pulse oximetry > or = 95%; n = 42) or NPPV by face mask plus standard medical therapy (n = 39). MAIN OUTCOME MEASURES: Rates of reintubation, duration of mechanical ventilation, lengths of ICU and hospital stay, and hospital mortality. RESULTS: Comparing the NPPV group with the standard therapy group, there was no difference in the rate of reintubation (72% vs 69%; relative risk, 1.04; 95% confidence interval, 0.78-1.38) or hospital mortality (31% for both groups; relative risk, 0.99; 95% confidence interval, 0.52-1.91). Similarly, no difference was found in duration of mechanical ventilation or length of ICU or hospital stay. CONCLUSIONS: The addition of NPPV to standard medical therapy does not improve outcome in heterogeneous groups of patients who develop respiratory distress during the first 48 hours after extubation. PMID- 12076222 TI - A 23-year-old man with schizophrenia. PMID- 12076223 TI - A 40-year-old woman who noticed a medication error, 1 year later. PMID- 12076224 TI - Preventing cardiac events and restenosis after percutaneous coronary intervention. PMID- 12076225 TI - Dietary intake of antioxidants and risk of Alzheimer disease: food for thought. PMID- 12076230 TI - Statin therapy in older persons: pertinent issues. PMID- 12076231 TI - Reduction of left ventricular hypertrophy after exercise and weight loss in overweight patients with mild hypertension. AB - BACKGROUND: Hypertrophy and concentric remodeling of the left ventricle are important manifestations of hypertension that are associated with increased morbidity and mortality. Although lifestyle interventions are efficacious in lowering blood pressure, evidence that they have a beneficial effect on target organs has been lacking. OBJECTIVE: To assess the effects of regular aerobic exercise or exercise plus weight management counseling on left ventricular mass and geometry in overweight, sedentary men and women with high-normal or mildly elevated blood pressure. METHODS: Eighty-two participants in a randomized, controlled trial were assigned to supervised aerobic exercise only, a behavioral weight management program that included exercise, or a waiting-list control group for 6 months. Blood pressure and echocardiographic measures of left ventricular structure were measured at baseline and at the conclusion of the treatment phase. RESULTS: The 45 women and 37 men had a mean +/- SD age of 47 +/- 9 years and had a mean +/- SD blood pressure of 140 +/- 10/93 +/- 5 mm Hg. Blood pressure fell by 7/6 mm Hg in the weight management group and by 3/4 mm Hg in the aerobic exercise group. In association with these decreases in blood pressure, participants in the intervention groups exhibited significant decreases in left ventricular relative wall thickness (P =.003), posterior wall thickness (P =.05), and septal thickness (P =.004) and a trend toward a decrease in indexed left ventricular mass (P =.08) relative to the control group. CONCLUSIONS: In a cohort of overweight, sedentary men and women, exercise and weight loss reduced blood pressure and induced favorable changes in left ventricular structure. PMID- 12076232 TI - Causes of physician delay in the diagnosis of breast cancer. AB - BACKGROUND: Understanding sources of physician delay in diagnosis of breast cancer will assist efforts to expedite diagnosis. OBJECTIVE: To test whether increased reliance on screening mammography has affected causes of physician delay in diagnosis of breast cancer. DESIGN: Survey of delays in a case series. SETTING: Practice specializing in breast diseases in a region with high use of screening mammography. PATIENTS: Four hundred thirty-five consecutive patients treated for 454 breast cancers of any stage. INTERVENTION: Customary patient care. MAIN OUTCOME MEASURES: Whether delay was related to how cancer was identified, patient age, individual cancer characteristics (such as tumor type), mammography reports, or physician expertise. RESULTS: Twenty-one women (5%) were inappropriately reassured that a malignant lump was benign without biopsy, 14 women (3%) had a misread mammogram, 4 women (1%) had a misread pathologic finding, and 5 women (1%) had cancer missed by a poorly performed fine-needle aspiration biopsy. Delay was associated with a benign mammography report (relative risk, 10.8; 95% confidence interval, 5.1-22.8), a woman finding her own mass (relative risk, 3.3; 95% confidence interval, 1.8-6.2), and current hormone replacement therapy (relative risk, 3.1; 95% confidence interval, 1.2-8.5). CONCLUSIONS: The leading cause of physician delay in diagnosis of breast cancer continues to be inappropriate reassurance that a mass is benign without biopsy. Reducing delay in diagnosis will require less willingness to rely on clinical examination to decide that a mass is benign, less reliance on benign mammography reports to decide not to biopsy a mass, and a requirement that fine-needle aspiration biopsy be done by persons with demonstrated competence for the procedure. PMID- 12076233 TI - Sildenafil for male erectile dysfunction: a systematic review and meta-analysis. AB - OBJECTIVE: To determine the efficacy and safety of sildenafil citrate in the treatment of male erectile dysfunction. DATA SOURCES: The MEDLINE, HealthSTAR, Current Contents, and Cochrane Library databases (January 1, 1995, through December 31, 2000); bibliographies of retrieved articles and review articles; conference proceedings abstracts; the Food and Drug Administration Web site; and the manufacturer. STUDY SELECTION: Trials were eligible if they included men with erectile dysfunction, compared sildenafil with control, were randomized, were of at least 7 days' duration, and assessed clinically relevant outcomes. DATA EXTRACTION: Two reviewers independently evaluated study quality and extracted data in a standardized fashion. DATA SYNTHESIS: Twenty-seven trials (6659 men) met the inclusion criteria. In results pooled from 14 parallel-group, flexible as needed dosing trials, sildenafil was more likely than placebo to lead to successful sexual intercourse, with a higher percentage of successful intercourse attempts (57% vs 21%; weighted mean difference, 33.7; 95% confidence interval [CI], 29.2-38.2; 2283 men) and a greater percentage of men experiencing at least 1 intercourse success during treatment (83% vs 45%; relative benefit increase, 1.8; 95% CI, 1.7-1.9; 2205 men). In data pooled from 6 parallel-group, fixed-dose trials, efficacy appeared slightly greater at higher doses. Treatment response appeared to vary between patient subgroups, although relative to placebo, sildenafil significantly improved erectile function in all evaluated subgroups. In trials with parallel-group design and flexible dosing, men randomized to receive sildenafil were less likely than those receiving placebo to drop out for any reason and no more likely to drop out due to an adverse event or laboratory abnormality. Specific adverse events with sildenafil included flushing (12%), headache (11%), dyspepsia (5%), and visual disturbances (3%); all adverse events were significantly less likely to occur with placebo. Sildenafil was not significantly associated with serious cardiovascular events or death. CONCLUSIONS: Sildenafil improves erectile function and is generally well tolerated. Treatment response seems to vary between patient subgroups, although sildenafil has greater efficacy than placebo in all evaluated subgroups. PMID- 12076234 TI - Willingness to pay for complete symptom relief of gastroesophageal reflux disease. AB - BACKGROUND: Over $6 billion per year is spent on prescription medication for gastroesophageal reflux disease (GERD). This study is an economic analysis of patients' willingness to pay for a prescription medication that offers complete relief of GERD symptoms. METHODS: The study was a cross-sectional, nonrandomized design recruiting patients from 5 clinical sites. A computer-administered discrete-choice questionnaire was used to explore patients' willingness to pay for various attributes (time to relief, amount of relief, side effects, and out of-pocket cost) associated with GERD treatment. Patients chose between 2 different combinations of attributes by indicating which scenario they preferred. Data were gathered on health status, health-related quality of life, and sociodemographic characteristics. RESULTS: Two hundred five patients completed the discrete-choice questionnaire with a consistency rate of 99.5%. All attributes were relevant to patient decision making. Respondents were willing to pay up to $182 to obtain complete relief in a short period of time without side effects. Patients with less severe GERD symptoms were willing to pay more to avoid side effects ($58.25 vs $38.43). Older patients were less willing to pay for better relief than younger patients. CONCLUSIONS: Results demonstrate that patients are willing to pay more per month for a medication that provides more complete and faster relief from GERD symptoms. This information can guide clinicians and formulary committees in evaluating optimal treatment for GERD. PMID- 12076235 TI - The increasing incidence of coronary artery disease and cardiovascular risk factors among a Southwest Native American tribe: the White Mountain Apache Heart Study. AB - BACKGROUND: Rates of cardiovascular disease and its risk factors seem to be increasing in American Indian populations, yet these changes have received little documentation. OBJECTIVES: To evaluate incidence rates of coronary artery disease, acute myocardial infarction, and cardiac events during a 10-year period (1987-1996); to assess cardiac risk factors for an American Indian tribe in Arizona. METHODS: A retrospective medical chart review was performed for tribal members from January 1, 1987-December 31, 1996. Patient records with even minor indications of coronary disease were reviewed independently by 2 cardiologists of the Native American Cardiology Program. Multiple databases were reviewed in an effort to find all diagnoses, and incidence rates were calculated and analyzed for increasing trends. Cardiac risk factors were assessed in a population convenience sample. RESULTS: From 1987 through 1996, the number of incident cases increased from 3 to 18 for coronary artery disease, 1 to 10 for acute myocardial infarction, and 3 to 26 for cardiac events. Statistically significant increasing trends were calculated for each. Of our youthful convenience sample, 49% had 2 or more cardiac risk factors. CONCLUSIONS: This study confirms increasing rates of coronary artery disease and its comorbidities in this American Indian population and demonstrates high prevalence of cardiovascular risk factors among tribal members without extant coronary disease. This suggests that coronary disease will likely continue to increase as this population ages. To prevent such increases, culturally appropriate, aggressive preventive interventions are needed. PMID- 12076236 TI - Plasma total homocysteine and hospitalizations for cardiovascular disease: the Hordaland Homocysteine Study. AB - BACKGROUND: Elevated total plasma homocysteine (tHcy) level is a risk factor for occlusive disease in the coronary, cerebral, and peripheral vessels and is related to several lifestyle factors associated with cardiovascular disease (CVD). OBJECTIVE: To examine the association of a single tHcy measurement on subsequent hospitalizations due to CVD. METHODS: A population-based prospective cohort study was conducted from April 1, 1992, to May 31, 1998 (mean follow-up, 5.3 years) in western Norway. The study included 17 361 individuals aged 40 to 42 or 65 to 67 years at baseline. Main outcome measure was CVD as the main hospital discharge diagnosis or coronary revascularization procedures (denoted "CVD hospitalizations") during follow-up (n = 1275). RESULTS: At baseline, participants with preexisting CVD had higher mean tHcy values than individuals without CVD. Risk of CVD hospitalizations increased significantly with increasing baseline tHcy only in the oldest age group. Here, multiple risk factor-adjusted hospitalization rate ratios in 5 tHcy categories (<9, 9-11.9, 12-14.9, 15-19.9, and >or=20 micromol/L [to convert tHcy to milligrams per liter, divide by 7.397]) were as follows: 1 (reference level), 1.00, 1.34, 1.67, and 1.94, respectively (P for trend <.001). The relation between tHcy level and CVD hospitalizations was significantly stronger among individuals with preexisting CVD than those without (hospitalization rate ratio per 5-micromol/L tHcy increment, 1.29 vs 1.10; P for interaction,.02). CONCLUSIONS: Plasma tHcy level is a strong predictor of CVD hospitalizations only in elderly individuals, and especially among those with preexisting CVD. Our findings are compatible with the theory that tHcy interacts with conventional CVD risk factors to provoke the acute event of CVD. PMID- 12076237 TI - Nut consumption and decreased risk of sudden cardiac death in the Physicians' Health Study. AB - BACKGROUND: Dietary nut intake has been associated with a reduced risk of coronary heart disease mortality; however, the mechanism is unclear. Since components of nuts may have antiarrhythmic properties, part of the benefit may be due to a reduction in sudden cardiac death. METHODS: We prospectively assessed whether increasing frequency of nut consumption, as ascertained by an abbreviated food frequency questionnaire at 12 months of follow-up, was associated with a lower risk of sudden cardiac death and other coronary heart disease end points among 21 454 male participants enrolled in the US Physicians' Health Study. Participants were followed up for an average of 17 years. RESULTS: Dietary nut intake was associated with a significantly reduced risk of sudden cardiac death after controlling for known cardiac risk factors and other dietary habits (P for trend,.01). Compared with men who rarely or never consumed nuts, those who consumed nuts 2 or more times per week had reduced risks of sudden cardiac death (relative risk, 0.53; 95% confidence interval, 0.30-0.92) and total coronary heart disease death (relative risk, 0.70; 95% confidence interval, 0.50-0.98). In contrast, nut intake was not associated with significantly reduced risks of nonsudden coronary heart disease death or nonfatal myocardial infarction. CONCLUSION: These prospective data in US male physicians suggest that the inverse association between nut consumption and total coronary heart disease death is primarily due to a reduction in the risk of sudden cardiac death. PMID- 12076238 TI - Rapid down-regulation of thyroid hormones in acute myocardial infarction: is it cardioprotective in patients with angina? AB - BACKGROUND: In severe illness of any cause, down-regulation of the thyroid hormone system may occur. How this affects patients with acute myocardial infarction (AMI) is largely unknown. OBJECTIVE: To investigate changes in serum levels of the thyroid hormones during AMI and their association with cardiac function and outcome. METHODS: Forty-seven consecutive euthyroid patients with AMI were studied prospectively during the first 5 days and again 6 and 12 weeks later. Time from pain onset was used in all analyses. RESULTS: The thyroid hormone system was rapidly down-regulated with maximal changes 24 to 36 hours after onset of symptoms. The mean level of the hormone total triiodothyronine (T3) decreased 19% (P =.02), the inactive metabolite reverse T3 (rT3) levels increased 22% (P =.01), and thyrotropin levels declined 51% (P<.001) between the first 6-hour and the 24- to 36-hour period. The prohormone free thyroxine was largely unchanged. Patients with poor heart function or more intense inflammatory reaction showed more pronounced down-regulation of the thyroid system. No correlation was found with cardiac enzymes. Patients with prior angina pectoris had lower T(3) levels in early samples, smaller infarctions, and higher levels of C-reactive protein and the proinflammatory cytokine interleukin 6 on admittance. Peak levels of interleukin 6 correlated negatively with T3 (P =.005) and positively with rT3 (P<.05), suggesting that down-regulation before AMI may be cardioprotective. However, mortality was high among patients with the most pronounced thyroid level depression, indicating that down-regulation after AMI may be maladaptive. CONCLUSIONS: The thyroid hormone system is rapidly down regulated in AMI. This may be beneficial during acute ischemia. Patients with angina had higher levels of interleukin 6 and C-reactive protein and more depressed thyroid hormone system in early samples. Thyroid level depression in patients with angina may possibly have been present before the infarction process started. This novel finding needs further evaluation in large studies to sort out cause-and-effect relationships. PMID- 12076239 TI - Therapy with hydroxymethylglutaryl coenzyme a reductase inhibitors (statins) and associated risk of incident cardiovascular events in older adults: evidence from the Cardiovascular Health Study. AB - BACKGROUND: Recommendations to treat older adults with hydroxymethylglutaryl coenzyme A reductase inhibitors (statins) for the primary prevention of coronary heart disease events are supported by a single clinical trial restricted to adults 73 years or younger with low levels of high-density lipoprotein cholesterol. METHODS: We investigated the association of statin use with incident cardiovascular disease and all-cause mortality during up to 7.3 years' follow-up of 1250 women and 664 men from the Cardiovascular Health Study. Study participants were 65 years and older and free of cardiovascular disease at baseline. They received drug therapy to lower cholesterol levels at baseline or no treatment with a recommendation for therapy according to the National Cholesterol Education Program guidelines. Use of these drugs was assessed annually. We used proportional-hazards models to calculate hazard ratios (HRs) and 95% confidence intervals (CIs), adjusted for confounding variables. RESULTS: We found 382 incident cardiovascular events (159 myocardial infarctions, 159 strokes, and 64 deaths due to coronary heart disease) and 362 total deaths from June 1, 1989, to May 31, 1997. Compared with no use of drugs to lower cholesterol levels, statin use was associated with decreased risk of cardiovascular events (multivariate HR, 0.44; 95% CI, 0.27-0.71) and all-cause mortality (HR, 0.56; 95% CI, 0.36-0.88). Similar associations were observed among participants 74 years or older at baseline. CONCLUSIONS: Use of statins was associated with decreased risk of incident cardiovascular events among elderly adults. These findings lend support to the National Cholesterol Education Program guidelines, which recommend therapy for the lowering of cholesterol levels for older adults with hypercholesterolemia. PMID- 12076240 TI - Association of kidney function with anemia: the Third National Health and Nutrition Examination Survey (1988-1994). AB - BACKGROUND: Kidney failure is known to cause anemia, which is associated with a higher risk of cardiac failure and mortality. The impact of milder decreases in kidney function on hemoglobin levels and anemia in the US population, however, is unknown. METHODS: We analyzed a population-based sample of 15419 participants 20 years and older in the Third National Health and Nutrition Examination Survey, conducted from 1988 to 1994. RESULTS: Lower kidney function was associated with a lower hemoglobin level and a higher prevalence and severity of anemia below, but not above, an estimated glomerular filtration rate (GFR) of 60 mL/min per 1.73 m(2). Adjusted to the age of 60 years, the predicted median hemoglobin level among men (women) decreased from 14.9 (13.5) g/dL at an estimated GFR of 60 mL/min per 1.73 m(2) to 13.8 (12.2) g/dL at an estimated GFR of 30 mL/min per 1.73 m(2) and to 12.0 (10.3) g/dL at an estimated GFR of 15 mL/min per 1.73 m(2). The prevalence of anemia (hemoglobin level <12 g/dL in men and <11 g/dL in women) increased from 1% (95% confidence interval, 0.7%-2%) at an estimated GFR of 60 mL/min per 1.73 m(2) to 9% (95% confidence interval, 4%-19%) at an estimated GFR of 30 mL/min per 1.73 m(2) and to 33% (95% confidence interval, 11%-67%) at an estimated GFR of 15 mL/min per 1.73 m(2) among men and to 67% (95% confidence interval, 30%-90%) at an estimated GFR of 15 mL/min per 1.73 m(2) among women. An estimated GFR of 15 to 60 mL/min per 1.73 m(2) was present in 4% of the entire population and in 17% of the individuals with anemia. CONCLUSION: Below an estimated GFR of 60 mL/min per 1.73 m(2), lower kidney function is strongly associated with a higher prevalence of anemia among the US adult population. PMID- 12076242 TI - Brugada syndrome: an unusual cause of convulsive syncope. AB - A patient who presented with a new apparent seizure was found to have abnormal electrocardiographic findings, with classic features of the Brugada syndrome. He had spontaneous episodes of nonsustained ventricular tachycardia, easily inducible ventricular fibrillation at electrophysiological study in the absence of structural heart disease, and a negative neurological evaluation. These findings suggested that sustained ventricular arrhythmias known to be associated with the Brugada syndrome and resultant cerebral hypoperfusion, rather than a primary seizure disorder, were responsible for the event. Patients with the Brugada syndrome often present with sudden death or with syncope resulting from ventricular arrhythmias. In consideration of its variability in presentation sometimes mimicking other disorders, primary care physicians and internists should be aware of its often transient electrocardiographic features. PMID- 12076241 TI - Alveolar and postcranial bone density in postmenopausal women receiving hormone/estrogen replacement therapy: a randomized, double-blind, placebo controlled trial. AB - BACKGROUND: We conducted a 3-year, double-blind, randomized, placebo-controlled study to determine whether the positive effects of hormone/estrogen replacement therapy (H/ERT) on postcranial bone density are accompanied by similar positive effects on oral bone mass. METHODS: A total of 135 postmenopausal women (aged 41 70 years) with no evidence of moderate or severe periodontal disease were randomized to receive daily oral conjugated estrogen (Premarin; 0.625 mg) alone or in combination with medroxyprogesterone acetate (Prempro; 0.625 and 2.5 mg, respectively) or placebo. All subjects received calcium carbonate (1000 mg/d) and cholecalciferol (400 [corrected] IU/d) supplements. The primary efficacy end points were the changes in alveolar crest height and alveolar bone density. Alveolar crest height was measured on bite-wing radiographs, and changes in alveolar bone mass were assessed by means of digital-subtraction radiography. Postcranial bone density was measured in the lumbar spine and left proximal femur by means of dual-energy x-ray absorptiometry. RESULTS: Hormone/estrogen replacement therapy significantly increased alveolar bone mass compared with placebo (+1.84% vs +0.95% [P =.04]), and tended to improve alveolar crest height (+4.83% vs +3.46% [P =.34]). Bone mineral density of the proximal femur significantly increased in the H/ERT compared with the placebo group (total proximal femur, +3.59% vs +0.22% [P =.001]; neck, +2.05% vs -0.34% [P =.02]; trochanter, +3.49% vs +0.08% [P<.001]), but not the lumbar spine (+1.01% vs +0.17% [P =.39]). Changes in alveolar bone mass correlated with bone density changes in the total femur (r = 0.28 [P =.02]) and femoral trochanter (r = 0.25 [P =.04]) in the H/ERT but not in the placebo group. CONCLUSIONS: Postcranial and oral bone mass were increased in postmenopausal women receiving H/ERT. Improvement in oral bone health constitutes an additional benefit of H/ERT. PMID- 12076243 TI - Massah and mechanisms. PMID- 12076244 TI - The effects of prayer: scientific study. PMID- 12076245 TI - Can the efficacy of prayer be tested? PMID- 12076246 TI - Must science and religion be separate? PMID- 12076247 TI - Research and religion. PMID- 12076249 TI - God, science, and intercessory prayer. PMID- 12076253 TI - Mast cells and angiogenesis. AB - Angiogenesis is tightly regulated by pro- and anti-angiogenic factors. Secreting mast cells are able to induce and enhance angiogenesis via multiple in part interacting pathways. They include mast cell-derived (i) potent pro-angiogenic factors such as VEGF, bFGF, TGF-beta, TNF-alpha and IL-8, (ii) proteinases and heparin, that release heparin-binding pro-angiogenic factors lodged on cell surfaces and in the extracellular matrix (ECM), (iii) histamine, VEGF, and certain lipid-derived mediators that induce microvascular hyperpermeability having pro-angiogenic effects, (iv) chemotactic recruitment of monocytes/macrophages and lymphocytes that are able to contribute with angiogenesis-modulating molecules, (v) activation of platelets that release pro angiogenic factors, (vi) activation of neighboring stationary non-mast cells, which secrete pro-angiogenic factors, ECM-degrading proteinases and stem cell factor which attracts, mitogenically stimulates and activates mast cells, (vii) auto- and paracrine stimulation of mast cells by stem cell factor, (viii) recruitment of mast cells by pro-angiogenic factors such as VEGF, bFGF and TGF beta. As a result of ECM-degradation and changes in the microenvironment following initial mast cell secretion, the mast cell populations may change significantly in number, phenotype and function. In tumor models, mast cells have been shown to play a decisive role in inducing the angiogenic switch which precedes malignant transformation. There is, moreover, strong evidence that mast cells significantly influence angiogenesis and thus growth and progression in human cancers. PMID- 12076251 TI - The cyclo-oxygenase-2 inhibitor celecoxib perturbs intracellular calcium by inhibiting endoplasmic reticulum Ca2+-ATPases: a plausible link with its anti tumour effect and cardiovascular risks. AB - Substantial evidence indicates that the cyclo-oxygenase-2 (COX-2) inhibitor celecoxib, a widely prescribed anti-inflammatory agent, displays anti-tumour effect by sensitizing cancer cells to apoptosis. As part of our effort to understand the mechanism by which celecoxib mediates apoptosis in androgen independent prostate cancer cells, we investigated its effect on intracellular calcium concentration ([Ca(2+)](i)). Digital ratiometric imaging analysis indicates that exposure of PC-3 cells to celecoxib stimulates an immediate [Ca(2+)](i) rise in a dose- and time-dependent manner. Kinetic data show that this Ca(2+) signal arises from internal Ca(2+) release in conjunction with external Ca(2+) influx. Examinations of the biochemical mechanism responsible for this Ca(2+) mobilization indicate that celecoxib blocks endoplasmic reticulum (ER) Ca(2+)-ATPases. Consequently, inhibition of this Ca(2+) reuptake mechanism results in Ca(2+) mobilization from ER stores followed by capacitative calcium entry, leading to [Ca(2+)](i) elevation. In view of the important role of Ca(2+) in apoptosis regulation, this Ca(2+) perturbation may represent part of the signalling mechanism that celecoxib uses to trigger rapid apoptotic death in cancer cells. This Ca(2+)-ATPase inhibitory activity is highly specific for celecoxib, and is not noted with other COX inhibitors tested, including aspirin, ibuprofen, naproxen, rofecoxib (Vioxx), DuP697 and NS398. Moreover, it is noteworthy that this activity is also observed in many other cell lines examined, including A7r5 smooth muscle cells, NIH 3T3 fibroblast cells and Jurkat T cells. Consequently, this Ca(2+)-perturbing effect may provide a plausible link with the reported toxicities of celecoxib such as increased cardiovascular risks in long term anti-inflammatory therapy. PMID- 12076254 TI - Postnatal development of beta-cells in rats. Proposed explanatory model. AB - The previously shown wave of beta-cell apoptosis and the apparent plateau in the beta-cell mass in the third week of life in rats are still unexplained events. Using a novel design-based stereological method we investigated the postnatal development of the beta-cell population in Sprague-Dawley rats. The total beta cell mass increased from postnatal day 4 until day 16, to be followed by a plateau until day 24, after which it increased further. This plateau was caused by beta-cell hypotrophia as well as decreased net beta-cell formation. The beta cell mass per unit body weight (the relative beta-cell mass) was five times higher at birth compared with the adult constant level that was reached at approximately 24 days of age. We propose an explanatory model for the postnatal development of the beta-cell population in rats. According to this model, beta cells in the early postnatal period are immature, i.e. are not susceptible to the mechanism that in later life maintains a constant relative beta-cell mass. Within the following weeks the number of mature beta-cells increases, and from approximately day 24 and onwards the beta-cell population is dominated by mature beta-cells that adjust to match the body weight, keeping a constant relative beta cell mass. Findings of an apoptotic wave, a plateau phase in the total beta-cell mass development, a period with beta-cell hypotrophia, and the disappearance of insulin-like growth factor II positive beta-cells at postnatal day 21 all fit well in the model. PMID- 12076252 TI - Requirement of the p38 mitogen-activated protein kinase signalling pathway for the induction of the 78 kDa glucose-regulated protein/immunoglobulin heavy-chain binding protein by azetidine stress: activating transcription factor 6 as a target for stress-induced phosphorylation. AB - Malfolded protein formation and perturbance of calcium homoeostasis results in the induction of the endoplasmic reticulum (ER) chaperone protein, namely the 78 kDa glucose-regulated protein (GRP78)/immunoglobulin heavy-chain binding protein. Various ER stress inducers can activate grp78, but signal transduction mechanisms are not well understood. We report in the present study that the induction of endogenous grp78 mRNA by the amino acid analogue azetidine (AzC) requires the integrity of a signal transduction pathway mediated by p38 mitogen-activated protein kinase (p38 MAPK). In contrast, induction of grp78 by thapsigargin that depletes the ER calcium storage can occur even when the p38 MAPK pathway is blocked. Treatment of cells with AzC results in the sustained activation of p38 MAPK. We identified an ER transmembrane activating transcription factor 6 (ATF6) as a target of p38 MAPK phosphorylation in AzC-treated cells. ATF6 undergoes proteolytic cleavage on AzC treatment, releasing a nuclear form that is an activator of the grp78 promoter. We show here that constitutively active mitogen activated protein kinase kinase 6, a selective p38 MAPK activator, enhances the ability of the nuclear form of ATF6 to transactivate the grp78 promoter. Our results provide direct evidence that different ER stress inducers use diverse pathways to activate grp78 and that in addition to activation by proteolytic cleavage, ATF6 undergoes specific ER stress-induced phosphorylation. We propose that phosphorylation of ATF6 is a novel mechanism for augmenting its potential as a transcription activator. PMID- 12076256 TI - Investigation of Candida dubliniensis in Candida spp.-positive hemocultures. AB - Candida dubliniensis is one of the Candida species which was first recognized in 1995. The yeast was misidentified because of its phenotypic similarities with Candida albicans. In this study, blood samples of patients from various departments at Ankara University Medical Faculty between January 1996 and September 2000 were investigated for distribution of Candida spp. and presence of C. dubliniensis. Ninety-eight culture positive fungi were included in the study. Phenotypic tests for identification of C. dubliniensis and tests for differentiation of the yeast from C. albicans, such as colony morphology on Staib agar, growth at 42 degrees C and 45 degrees C, beta-glucosidase activity and carbohydrate assimilation, were carried out. Sixty-four of the isolates produced germ tubes and chlamydospores, and none of them had the phenotypic characteristics of C. dubliniensis. Further large-scale studies of specific patient groups are necessary to reveal the etiologic importance of this yeast. PMID- 12076255 TI - T-cell-rich B-cell lymphoma - diagnostic and therapeutic aspects. AB - BACKGROUND: Morphologically, T-cell-rich B-cell lymphoma (TCRB-NHL) may be indistinguishable from Hodgkin's disease (HD). Immunophenotyping may be helpful in the separation of these entities. TCRB-NHL is occasionally misdiagnosed and treated as HD. However, information is limited regarding clinical characteristics and outcome of this patient population. Furthermore, knowledge concerning any association with Epstein-Barr virus (EBV) in TCRB-NHL, as well as the immunophenotype of reactive T-cells and the expression of T-cell intracellular antigen-1 (TIA-1), granzyme B (GrB) and the CD3-zeta-chain is limited. PATIENTS AND METHODS: We have re-evaluated 251 tumour biopsies from patients aged > or =15 years with HD diagnosed 1985-1994. Reclassification from HD to TCRB-NHL was done in 12 cases (5%). Six TCRB-NHL patients initially diagnosed and treated as B-NHL were also included. All TCRB-NHL biopsies were analysed for latent membrane protein 1 (LMP-1), CD4, CD8, CD56, CD57, TIA-1, GrB and CD3-zeta-chain. RESULTS: Twelve cases of TCRB-NHL were initially subclassified as HD (lymphocyte predominance 5, nodular sclerosis 3, and mixed cellularity 4). Of these 12 TCRB NHL patients, 6 were given radiotherapy alone, 5 MOPP/ABVD or similar combination chemotherapy, and one patient combined modality treatment. Male sex (p<0.05) and inguinal involvement (p<0.001) were significantly more frequent when TCRB-NHL patients receiving HD treatment (n=12) were compared with the remaining patients with confirmed (conf) HD, while no significant differences were seen with regard to stage, bone marrow infiltration, splenomegaly or cause-specific survival. Similar results were achieved when all TCRB-NHL patients (n=18) were compared to conf HD patients. Lymphoma cells in three samples stained positively for LMP-1. A decreased expression of CD3-zeta-chain was seen in 9/14 tumour biopsies. CONCLUSION: Immunohistochemistry makes it possible to identify cases of TCRB-NHL that are morphologically difficult to distinguish from HD. The outcome of TCRB NHL patients treated as having HD was comparable with that of the remaining HD population. PMID- 12076258 TI - No immunosuppressive effect of transfected annexin II. AB - Annexin II (AXII) was reported to suppress lymphoproliferation and immunoglobulin production. To investigate this further, the AXII-negative human B-lymphoma cell line Raji was transfected with AXII followed by transfection with p11. Stable transfectants were generated. In vitro immunological effects of Raji, Raji-AXII and Raji-AXII-p11 were compared by using irradiated cells as stimulators for PBM in mixed lymphocyte reactions or by adding the supernatants to lymphoproliferation or ELISPOT cultures. None of the assays provided evidence of significant immunosuppression by AXII. Thus, expression of AXII or AXII plus p11 does not by itself give a cell immunosuppressive ability. PMID- 12076257 TI - Bismuth-induced lysosomal rupture in J774 cells. AB - Bismuth-containing drugs have several applications, one being their use against Helicobacter pylori-associated peptic ulcers, and bismuth has been discovered in macrophages at the base and margins of peptic ulcers. In the present study, the autometallographic technique for the histochemical demonstration of bismuth was applied, showing that bismuth citrate-exposed J774 cells accumulate the metal in their lysosomes. Such accumulations resulted in lysosomal rupture - assayed by the acridine orange uptake technique and flow cytofluorometry - and ensuing apoptotic cell death. PMID- 12076259 TI - Synergistic effects of 1,25-Dihydroxyvitamin D3 and TGF-beta1 on the production of insulin-like growth factor binding protein 3 in human bone marrow stromal cell cultures. AB - 1,25-Dihydroxyvitamin D3 (calcitriol), transforming growth factor-beta (TGF beta), and insulin-like growth factors (IGFs) are all important bone regulatory factors known to affect proliferation and differentiation of human bone-forming cells (osteoblasts). We have previously shown that TGF-beta1 increased IGF-I and IGF-binding protein (IGFBP)-3 production in human bone marrow stromal (hMS) osteoblast progenitors and calcitriol stimulated IGFBP-3 and IGFBP-4 production. As interaction between signaling pathways of these factors has been reported, the present study aimed at examining the concerted actions on components of the IGF system. We report that co-treatment with TGF-beta1 and calcitriol resulted in a synergistic increase in IGFBP-3 production, thereby suggesting that the effects of these factors on hMS osteoblast differentiation may involve the observed increase in IGFBP-3. PMID- 12076260 TI - Uncoupling activation-induced modulation of CD16 and CD69 in CD56+ cells during AIDS. AB - The immune system of HIV+ patients is chronically activated, which has been associated with a detrimental effect on both innate and acquired immunity during AIDS. We analyzed the expression and modulation of the triggering markers CD69 and CD16 in CD56+ cells from 18 asymptomatic HIV+ individuals and 8 AIDS patients, compared with 21 seronegative subjects. We observed a diminished PMA induced CD16 downregulation in AIDS patients (p<0.01), associated with low numbers of CD4+ cells (p<0.02). Furthermore, an enhanced unstimulated expression of CD69 in asymptomatic HIV+ patients (p<0.05) was shown. AIDS patients could not efficiently upregulate PHA-dependent CD69 expression (p<0.05), which correlated with low CD4+ counts (p< 0.05). These abnormalities in CD16 and CD69 modulation were recorded in patients under highly active antiretroviral therapy (HAART). Our results demonstrate an altered modulation of two functionally relevant receptors in CD56+ cells from AIDS patients, contributing to our understanding of the immunopathogeny of NK cell dysfunction during disease progression. PMID- 12076262 TI - Infectious salmon anaemia virus. AB - Infectious salmon anaemia virus (ISAV) is a commercially important orthomyxovirus causing disease in farmed Atlantic salmon. The cumulative mortality in a net pen during an outbreak may vary from insignificant to more than 90%. The infection is spread by management activity such as well-boat traffic, but possibly also through contact with wild fish. In many of its aspects, including the structure of the virus particle and replication strategy, the ISAV is similar to the influenza viruses. Variations between ISAV and the influenza viruses can mostly be related to differences in the temperature at which replication occurs and the immune response of their respective host animals. ISAV shows both haemagglutinating and receptor-destroying activity. The variability of the ISAV haemagglutinin molecule is concentrated around a small domain close to the transmembrane region. The function of this variable region is unknown, but it may be related to a recent or ongoing crossing of a species barrier. Alignment studies based on genetic data indicate that the phylogenetic relationship to the influenza viruses is distant, and that ISAV therefore could possibly warrant a new genus within Orthomyxoviridae. PMID- 12076261 TI - Hypoxia and reoxygenation of brain microvascular smooth muscle cells in vitro: cellular responses and expression of cerebral amyloid angiopathy-associated proteins. AB - Hypoxia is known to cause complex cascades of physiological, biochemical, and morphological changes in the brain. Cerebral microvascular smooth muscle cell (MV SMC) damage may occur following hypoxic conditions and lead to SMC dysfunction. However, little is known about the exact cellular and molecular responses of these cells to hypoxia. To partly address these questions, MV-SMC were isolated from human brain, cultured and placed in conditions of ambient hypoxia (H) and hypoxia followed by reoxygenation (H/R). Cell morphology, proliferation, and the expression of amyloid precursor protein (APP) and cystatin C peptide were investigated and compared (after induction of hypoxia) between cerebral MV- and human aortic SMC. Our results show that MV-SMC proliferation was inhibited after 48 h of hypoxia and H/R, whereas aortic SMC proliferation was stimulated after 48 h of hypoxia and H/R. Hypoxia and H/R induced an increase of intracellular APP and cystatin C expression in both types of SMC, though the effect of H and H/R on APP upregulation was quantitatively more robust in MV-SMC than aortic SMC. Patterns of hypoxia-induced APP upregulation in SMC differed significantly from those found in cultured neuronal cells (PC12, NT2). These results suggest that hypoxia and H/R-induced APP and cystatin C upregulation appear to occur independently of the inhibition of cerebral MV-SMC proliferation. Overexpression of APP and cystatin C in response to hypoxia may thus represent an initiating event in the pathogenesis of amyloid angiopathy, or mediate progression of this microvascular lesion. PMID- 12076263 TI - Superficial collagenous fibroma: immunohistochemical, ultrastructural, and flow cytometric study of three cases, including one pemphigus vulgaris patient with a dermal mass. AB - Collagenous fibroma (desmoplastic fibroblastoma) is an extremely rare benign soft tissue tumor of fibroblastic origin. The majority of reported cases have been located in the deep subcutis, fascia, aponeurosis, or skeletal muscle of the extremities, limb girdles, or head and neck regions. There has been no mention of underlying diseases in patients who developed this tumor. We here report an additional three cases of superficial collagenous fibroma, one of which was a dermal lesion occurring in the abdomen of a 26-year-old male patient with a 5 year history of pemphigus vulgaris prior to development of the tumor. To the best of our knowledge, an association between collagenous fibroma and pemphigus vulgaris has not previously been reported. The remaining two tumors were located in the superficial subcutaneous tissue of the infrascapular area and right foot, respectively. There was no tumor recurrence or metastasis during follow up of 18, 25, and 47 months, respectively. All three tumors were well-circumscribed and unencapsulated without infiltrating borders. Histologically, the common denominator of all three cases was paucicellular proliferation of spindle or stellate fibroblasts enmeshed within an extensively collagenous background. Immunohistochemically, there was diffuse strong staining for vimentin and intense focal reaction for smooth muscle actin in two tumors tested. Electron microscopy revealed features consistent with a fibroblastic or myofibroblastic lineage. Flow cytometry in two cases demonstrated a diploid DNA content with low S-phase fractions, which correlated with minimal MIB-1 nuclear labeling (less than 1%) and benign behavior of this entity. PMID- 12076264 TI - Conditions influencing the in vitro antifungal activity of lactoferrin combined with antimycotics against clinical isolates of Candida. Impact on the development of buccal preparations of lactoferrin. AB - Lactoferrin, an iron-binding glycoprotein, is a potential agent for the treatment of oropharyngeal Candidiasis. The aim of the present study was to test the capability of lactoferrin, combined or not combined with conventional antifungal agents, to inhibit the growth of different Candida species under various experimental conditions to be of guidance in the development of a suitable pharmaceutical formulation containing lactoferrin. The anti-Candida activities of lactoferrin were considerably higher using RPMI instead of SLM as assay medium. They were moreover increased by raising the medium pH from 5.6 to 7.5. With the 'standard' antifungal agent fluconazole similar results were found as for lactoferrin, but the medium type and pH did not affect MIC values of amphotericin B. The addition of saliva to medium did not reduce the antifungal activities of the individual compounds. Synergistic inhibitory effects on Candida growth were found for combinations of lactoferrin and fluconazole or amphotericin B, irrespective of the medium type and pH, or the addition of saliva. This indicates that for treatment of oral Candidiasis a formulation containing lactoferrin seems appropriate; results may be optimized if the formulation is provided with buffer capacity to attain pH 7.5 in the mucosal fluid. The synergistic effects between lactoferrin and 'standard' antifungals indicate that combinations should be considered in such a formulation. PMID- 12076265 TI - Effects of magnetic cell separation on monocyte adhesion to endothelial cells under flow. AB - Studies on monocyte adhesion are frequently limited by spontaneous changes of CD11b and CD62L during cell purification. Most isolation protocols for flow cytometric analysis that overcome this problem cannot be used when large numbers of living cells are needed for functional adhesion assays. This study investigated whether magnetic cell separation of monocytes with a paramagnetic bead against CD33 is a feasible method combining high yield with a low degree of spontaneous activation. As determined by flow cytometry, isolation of magnetically tagged monocytes at 4 degrees C did not alter the expression of CD11b and CD62L when compared to whole blood controls. Warming the cells slowly to room temperature immediately before starting the adhesion assay in a parallel plate flow chamber at 37 degrees C prevented further upregulation of adhesion molecules due to rewarming. When adhesion of magnetically tagged monocytes was compared with untouched monocytes that had been isolated via depletion of contaminating leukocytes, videomicroscopy showed that labelling CD33 neither affected rolling nor firm adhesion to human umbilical venous endothelial cells under flow. Finally, the subsequent upregulation of tissue factor expression on adherent monocytes indicates that magnetically separated monocytes responded properly to activating stimuli during cell adhesion. We conclude that magnetic cell separation via CD33 represents a feasible method for cell separation whenever large numbers of non-activated monocytes are needed for adhesion assays under flow. PMID- 12076266 TI - Low prevalence of Bartonella henselae infections in Norwegian domestic and feral cats. AB - Bartonella henselae is the causative agent of cat scratch disease (CSD). This clinical entity is very rarely encountered in human medical practice in Norway. B. henselae infections including bacteraemia in cats have been frequently reported. The objective of the present study was to investigate the seroprevalence rate and the degree of B. henselae bacteraemia in Norwegian domestic and feral cats. One hundred cats investigated at a small animal veterinary practice in the middle of Norway were included in the study. Blood collected in Isolator blood-lysis tubes and lysates of erythrocytes after freezing and thawing were cultured. PCR analysis of whole blood was also performed. Serology was performed by indirect fluorescence assay (IFA) and enzyme immunoassay (EIA) using immobilised B. henselae Houston-1 strain as antigen. None of the 100 cats investigated was found to be bacteraemic. All 100 cats were seronegative when analysed by IFA; one cat was positive by EIA. The discrepancy between IFA and EIA of this particular cat is probably due to cross-reactive antibodies. Contrary to findings reported from several geographic regions, B. henselae infections in Norwegian cats appear to be virtually absent. This in turn may explain why CSD has not been reported in human medical practice in Norway. PMID- 12076267 TI - Molecular heterogeneity of fluconazole-resistant and -susceptible oral Candida albicans isolates within a single geographic locale. AB - The emergence of drug-resistant Candida albicans in immunocompromised patients is common. A disconcerting aspect of this phenomenon is the rapid emergence of C. albicans strains that are resistant to a widely used azole drug, fluconazole (FLZ). To understand the origin of FLZ-resistant yeast isolates, we investigated molecular profiles of 20 geographically related oral C. albicans isolates using three genotyping methods: randomly amplified polymorphic DNA-PCR, with six different primers (OBU1, OBU2, OBU3 RSD6, RSD11 and RSD12); electrophoretic karyotyping by pulsed-field gel electrophoresis; and HinfI restriction fragment analysis. Of the 20 isolates studied, 10 were FLZ- resistant and originated from patients with oral candidosis with a history of FLZ therapy, and the remainder were FLZ susceptible from individuals with oral candidosis, but without a history of FLZ therapy. A composite genotype was generated for each strain by combining molecular types derived from the three independent molecular methods. The composite profiles indicated genetic diversity amongst both the FLZ-resistant as well as -sensitive isolates, and no specific features emerged distinguishing the drug-resistant and -sensitive groups. These observations cast doubt on the theory of a clonal origin of FLZ-resistant C. albicans isolates. PMID- 12076268 TI - Follicular hyperplasia presenting with a marginal zone pattern in a reactive lymph node lesion. AB - Histologically, the marginal zone pattern of the lymph node is characterized by lymphoid follicles with three distinct layers. The inner layer is composed of follicular center zones, the middle layer of darkly stained mantle zones, and the outer layer of marginal zones. However, the marginal zone pattern is rarely seen in reactive lymph nodes except for mesenteric lymph nodes. We describe the clinicopathologic, immunohistochemical and genotypic findings of six cases of reactive follicular hyperplasia exhibiting the marginal zone pattern. The patients comprised three males and three females (age range 24 to 63 years; medium 56 years). Follow-up data were obtained from five patients. None of them developed malignant lymphomas during the follow-up period of from 5 to 204 months (median 68 months). Histologically, the lesion was characterized by numerous lymphoid follicles and partial distortion of lymph node structure. Varying degrees of progressive transformation of the germinal center (PTGC) were noted in the four cases. The marginal zone pattern was observed in some or most of the lymphoid follicles including PTGC. The marginal zone B cells were small to medium sized lymphocytes with round or slightly indented nuclei and a broad rim of pale cytoplasm. Some of them had a monocytoid appearance. They were CD20+, CD79a+, sIgM+/-, sIgD-, CD5-, CD10-, CD21-, CD23-, CD43-, CD45RO-, Bcl-6-, cyclin D1-, EMA- and p53-. A portion of them were Bcl-2 positive. Occasional large lymphoid cells with round or indented nuclei and moderate amounts of cytoplasm were observed in the marginal zone in four cases. These large lymphoid cells were usually CD20 positive, but Bcl-6 negative. A small number of them contained polytypic intracytoplasmic immunoglobulins. The polytypic nature of B lymphocytes was demonstrated by immunohistochemistry and polymerase chain reaction. Recognition of unusual marginal zone hyperplasia in reactive lymph node lesions is important to avoid confusion with nodal involvement in various low-grade B cell lymphomas presenting a marginal zone distribution pattern. PMID- 12076269 TI - Phenotypic and genotypic characterisation of blood isolates of coagulase-negative staphylococci in the newborn. AB - Coagulase-negative staphylococci (CNS) are the leading cause of late-onset sepsis in newborns (>72 h of age). Our aim was to determine whether phenotypic and/or genotypic differences existed between blood isolates of CNS regarded as inducers of sepsis or as contaminants. Ninety-seven bloodisolates of CNS recovered from newborns at the neonatal intensive care unit, Orebro, Sweden in 1983-1997 were analysed. Twenty-nine of them (30%) were classified as sepsis isolates and 68 (70%) as contaminants. The most prevalent species was Staphylococcus epidermidis (n=59). Staphylococcus haemolyticus (n=16) was most often isolated from newborns with the lowest gestational age and birth weight. Biochemical typing using the Phene Plate system (PhP) and genotyping using pulsed-field gel electrophoresis (PFGE) showed that the S. epidermidis isolates regarded as inducers of sepsis (n=16) were more homogeneous than isolates considered contaminants (n=37). One main genotypic group, representing seven (44%) isolates, was identified among the sepsis isolates. Phenotypically the S. epidermidis sepsis isolates comprised three major clusters. In contrast, among the S. epidermidis contaminants, eight genotypic groups and two phenotypic clusters were identified. The dominating genotypic group among the sepsis isolates of S. epidermidis may represent strains with higher invasive capacity. PMID- 12076270 TI - Effects of different embedding gels on periostealchondrogenesis in vitro. AB - To develop a more useful organ culture model for periosteal chondrogenesis in vitro, we compared the effects of the embedding of explants in agarose versus collagen gels. Chondrogenic differentiation was examined by means of histological observation and in terms of the expression of mRNA encoding two cartilage markers, Type II collagen and Aggrecan. Periosteal explants were derived from the tibiae of rabbits. These explants were embedded in either agarose gel or collagen gel and cultured for 6 weeks. Histological examinations revealed that in the agarose gel, cells were neither present in the explants nor in the gel. In the collagen gel, cells migrated from the explanted tissues into the gel, and some cells were round. However, no explants showed safranin-O staining. Only 10% of 10 surviving explants in the agarose gel expressed the Type II collagen gene and the Aggrecan gene. On the other hand, of 25 surviving explants in the collagen gel, the expression of the Type II collagen gene was detected in 18 explants (72%) and that of the Aggrecan gene was detected in 21 explants (84%). In conclusion, we demonstrated that the periosteum exerts chondrogenesis in certain circumstances and its chondrogenesis is closely related to the culture material. PMID- 12076271 TI - Ets-1 and integrin beta3 for lung metastasis from colorectal cancer. AB - Ets-1 functions as tissue-specific transcription factor and plays an important role in cell proliferation, differentiation, angiogenesis, and apoptosis. To elucidate the involvement of Ets-1 in lung metastasis from colorectal cancer, immunohistochemical analysis was performed on 51 colorectal cancer patients (22 with lung metastasis and 29 with advanced colorectal cancer showing no recurrence for at least 5 years after surgery). Tumorous and stromal Ets-1 immunoreactivity was evaluated. Ets-1 protein was demonstrated within stromal fibroblasts, myofibroblasts and capillaries, and also within carcinoma cells. Stromal Ets-1 immunoreactivity in primary colorectal cancer was statistically correlated with lung metastasis (p<0.05). In primary colorectal cancer, a significant association was observed between stromal Ets-1 immunoreactivity and vascular integrin beta3 expression (p<0.01). In the cases with lung metastasis, vascular integrin beta3 index in lung metastases was significantly diminished compared with primary tumors or liver metastases (p<0.01). In contrast, stromal or tumorous Ets-1 expression did not change. In a multivariate model using logistic stepwise regression analysis, vascular integrin beta3 and stromal Ets-1 overexpression were significantly and independently related to lung metastasis. Stromal Ets-1 and/or vascular integrin beta3 may be useful markers for risk of lung metastasis from colorectal cancer. PMID- 12076272 TI - Outer membrane vesicles from Neisseria meningitidis. AB - Flow cytometry was used to study the expression of leukocyte adhesion molecules CD11a, CD11b, CD11c, CD14, and CD62L (L-selectin) and production of reactive oxygen species (ROS) in an ex vivo human whole-blood system stimulated with lipopolysaccharide-containing outer membrane vesicles (LPS-OMV) from N. meningitidis. Results demonstrated a dose-dependent increase in surface expression of CD11a, CD11b, CD11c and CD14 in granulocytes and monocytes (maximal at 30-120 min) upon OMV-LPS challenge, whereas CD62L expression was heavily downregulated (maximal at 30-120 min). The OMV-associated LPS was almost as potent (on a weight basis) as purified LPS from E. coli in inducing adhesion molecule modulation but the response was delayed. Upon stimulation with OMV-LPS or E. coli-LPS, the production of intracellular ROS increased in both granulocytes and monocytes when dihydroethidium (DHE, mainly reflecting superoxide anion) was used as a probe, whereas peroxynitrite production monitored with dihydrorhodamine 123 (DHR) was not significantly changed. The OMV-mediated modulation of leukocyte adhesion molecule expression and increased ROS production may certainly lead to increased entrapment of leukocytes in the microcirculation and contribute to untoward inflammatory reactions as seen in systemic meningococcal disease. PMID- 12076273 TI - Effect of intravenous immunoglobulins on in vitro immunoglobulin formation in patients with antibody immunodeficiency. AB - Seventeen patients with antibody immunodeficiency (9 subclass IgG immunodeficiencies, 8 common variable immunodeficiencies) and clinically unambiguous immunodeficiency symptomatology participated in the study with 14 healthy donors. The patients were given regular intravenous immunoglobulin (IVIG) infusions with Endobulin. Blood was collected before and 7 days after infusion of the usual IVIG dose. Mononuclear cells were isolated from peripheral blood (PBMC) of the patients by Ficoll-Paque gradient centrifugation. In order to monitor the ability to inhibit or activate polyclonal production of immunoglobulins in vitro, we stimulated PBMC with pokeweed mitogen (PWM) and with a mixture of pokeweed mitogen + concanavalin A (PWM+ConA). We found that an immunomodulatory effect of IVIG persists in vitro even one week after infusion. Polyclonally stimulated IgA and IgM production was suppressed by IVIG infusion mainly in patients with IgG subclass deficiency. The positive stimulatory effect of IVIG infusion on IgG production was confirmed. The IgG production increased in vitro after infusion in both groups of patients and was significantly higher than in healthy donors. Co stimulation of PWM-stimulated cells with ConA caused an inhibition of immunoglobulin release in normal healthy donors. The infusion supported the capability of ConA to inhibit IgG production in vitro in patients with IgG subclass deficiency, whereas an increase in IgG production with PWM+ConA stimulation after infusion was found in CVID patients. We assume that lymphocytes activated by ConA produce suppressive factors, which can be affected by the IVIG infusion and which can have both an immunostimulatory and an immunosuppressive effect. PMID- 12076274 TI - Reduced expression of both Bax and Bcl-2 is independently associated with lymph node metastasis in human breast carcinomas. AB - Imbalance between pro-apoptotic and anti-apoptotic proteins, causing altered apoptosis, may lead to tumour development and tumour progression, and reduced response to adjuvant therapy. In this study, we evaluated the expression patterns of Bcl-2, Bcl-xL, and Bax protein in 126 primary invasive breast carcinomas, and the association with other clinicopathological parameters. We used immunohistochemical methods to evaluate protein expression. Reduced expression of both Bax and Bcl-2 was associated with lymphnode metastases in univariate analyses (one-way ANOVA) as well as in multivariate analysis (binary logistic regression) (Bcl-2 p=0.003 univariate, p=0.01 multivariate, Bax p=0.05 univariate, p=0.03 multivariate). Bcl-2 overexpression showed an inverse association with cyclin A (p=0.05), while expression of Bcl-xL showed an association only with cyclin D3 (p=0.04). Bcl-xL expression also showed a highly significant association with oestrogen receptor status (p=0.009). Bcl-2 and Bcl xL showed an association with different D-type cyclins, indicating different pathways of pathogenesis. Expression of Bcl-2 was associated with better patient survival in univariate analysis (Kaplan meyer p=0.04), but lost its prognostic value in multivariate analysis (Cox regression p=0.2). PMID- 12076275 TI - The effects of enoxaparin on the reparative processes in experimental osteonecrosis of the femoral head of the rat. AB - The blood supply of one femoral head of 6-month-old rats was severed by incising the periosteum of the neck and cutting the ligamentum teres. The rats were killed on the 30th postoperative day and the femoral bones were obtained for semiquantification of the reparative processes in the necrotic heads. Fourteen rats were treated with enoxaparin and 14 untreated animals served as controls. Statistically, the amounts of necrotic bone in the epiphysis were less, the extent of remodeling of the femoral heads was milder, and the articular cartilage degeneration was slighter in the enoxaparin-treated than untreated rats. There was no significant difference in the quantities of newly formed bone in femoral heads of treated and untreated rats. These findings are in agreement with the known effects of unfractionated and low-molecular-weight heparins which enhance osteoclastic bone resorption and angiogenesis and decrease osteoblastic bone formation. The former activities, operative in minimizing the structural distortion of the femoral head, oppose the crucial event in the pathogenesis of post-osteonecrotic osteoarthritis. PMID- 12076277 TI - Isolation of exfoliated colonocytes from human stool as a new technique for colonic cytology. AB - Cell exfoliation in the gut is an important cell renewal mechanism. To approach its investigation we applied a novel immunomagnetic technique for isolation of exfoliated cells from human stool. Exfoliated colonocytes were isolated from 168 stool samples. The cells were assessed microscopically using conventional stains and immunohistochemistry. The technique allowed us to obtain well-preserved colonocytes displaying characteristic features of well-differentiated colonic epithelium and positive immunostaining for cytokeratin 5/8. No mucin-producing cells were found. Exfoliated cells did not produce inducible nitric oxide synthase, albeit cultured colon carcinoma cells HT-29 analysed in parallel showed strong immunostaining. Analysis of exfoliated cell numbers in consecutive stool samples from the same subjects revealed considerable interindividual variation. Overall exfoliated colonocyte numbers were relatively low, isolation being unaffected by addition during the procedure of excessive amounts of HT-29 cells. Apoptosis was extremely rare among exfoliated colonocytes. Well-preserved exfoliated colonocytes can be consistently isolated from human faeces using a simple procedure. Our findings suggest that the actual process of cell exfoliation in the human colon may be much less intense than is generally accepted. Exfoliated cell isolation from human stool constitutes a convenient non invasive approach that can be used for diagnostic and research purposes. PMID- 12076276 TI - Expression of HGF/SF and Met protein is associated with genetic alterations of VHL gene in primary renal cell carcinomas. AB - We analyzed the genetic alterations of VHL, HGF/SF, and Met genes and the expression pattern of HGF/SF and Met protein in 26 renal cell carcinomas (RCCs). We found five mutations of the VHL gene and frequent LOH (50%) only in non papillary clear cell RCC. We found six cases in which the CpG island of VHL was methylated. In addition, one missense mutation of the HGF/SF gene was detected in clear cell RCC. HGF/SF and Met protein were expressed in 84.6% and 80.7% of RCCs, respectively. All of the cases with the genetic alterations of VHL or HGF/SF demonstrated strong expression of HGF/SF and Met protein in RCC cells. Statistically, genetic alterations of VHL and HGF/SF were significantly correlated with HGF/SF and Met expression (Fisher's exact test, p=0.022 and p=0.0070). Thus, these results strongly suggest that the expression of HGF/SF and Met protein is closely associated with the genetic alterations of VHL and HGF/SF in primary RCCs. PMID- 12076278 TI - Epidemiological typing and prevalence of integrons in multiresistant Acinetobacter strains. AB - Seventy-seven Acinetobacter isolates were recovered from patients in a Korean hospital during the period from November to December 1998. The isolates were genotyped using randomly amplified polymorphic DNA (RAPD) analysis for epidemiological relationship, and investigated for antibiotic susceptibility and presence of integrons. Sixty-nine Acinetobacter baumannii isolates were distributed into five groups by RAPD profiles, with 5, 1, 60, 2 and 1 in each group. The major RAPD group of 60 isolates was further divided into six subgroups by antibiograms. Eight isolates belonging to Acinetobacter DNA group 13TU were distributed among six RAPD groups. Seventy-three of the Acinetobacter isolates were resistant to eight or more of the antibiotics tested. Integrase genes were detected in 66 of the 69 A. baumannii (96%) and in 5 of the 8 Acinetobacter DNA group 13TU isolates (63%). The intI1 and intI2 genes were found in 63 and 8 isolates, respectively. The intI3 gene was not detected. All integron-carrying isolates were resistant to multiple antibiotics. All strains isolated from more than one patient carried integrons. According to the results, the presence of integrons was significantly (p<0.01) associated with multiple antibiotic resistance and nosocomial spread in Acinetobacter strains. PMID- 12076279 TI - Solitary fibrous tumor of the abdominal wall: a report of two cases immunohistochemical, flow cytometric, and ultrastructural studies and literature review. AB - Solitary fibrous tumors have been described at many extrapleural sites in recent years. However, solitary fibrous tumors arising from somatic soft tissue occur only rarely and can pose problems in the differential diagnosis from other benign or malignant soft tissue tumors. The majority of solitary fibrous tumors occurring in the somatic soft tissue have been found in the extremities and limb girdles, and the head and neck regions. There have been only eight published cases located in the abdominal wall. We herein report two female patients who developed solitary fibrous tumors of the abdominal wall that were not in association with the underlying peritoneum. Histologically, both tumors were characterized by a variety of architectural patterns, alternating hypercellular and hypocellular areas, proliferation of plump spindle cells, thick keloid-like and/or amianthoid collagen bundles, and ectatic staghorn-like vessels. Both tumors showed a diffuse strong reaction for CD34 and vimentin as well as focal positivity for bcl-2 and smooth muscle actin. A striking predominance in females was found in a literature review of solitary fibrous tumors of the abdominal wall, contrasting with other somatic soft tissue sites which show an equal gender distribution. Interestingly, expression of estrogen but not progesterone receptor was observed in both tumors. Ultrastructurally, the tumor cells displayed features of fibroblasts with dilated branching rough endoplasmic reticulum (RER) and Golgi apparatus. Both tumors assayed by flow cytometry demonstrated a diploid DNA content with an S-phase fraction of 7.9% and 11.4%, respectively. At follow up, both patients were well without evidence of recurrence or metastasis after surgical excision. PMID- 12076280 TI - Classification, identification and subtyping of bacteria based on pyrosequencing and signature matching of 16S rDNA fragments. AB - The rapid identification of the etiological agent of microbial infections can bring about both clinical and financial benefits. Thus, fast and generally applicable classification methods are needed that will enable us to rapidly distinguish pathogenic bacteria from commensals or saprophytic bacteria found in the same habitat. We here show that provisional classification of bacterial isolates can be performed on a large scale based on 16S rRNA sequence comparisons using Pyrosequencing, a recently described real-time DNA sequence analysis technique, and the concept of signature matching. The probes we have developed, together with the new technology, will enable early diagnosis of specific pathogens, which is critical for the rational use of antimicrobial therapy in clinical medicine. PMID- 12076281 TI - Transfusion medicine illustrated: a microscopic atlas of thrombotic thrombocytopenic purpura. PMID- 12076282 TI - Thrombotic thrombocytopenic purpura and hemolytic uremic syndrome: will recent insight into pathogenesis translate into better treatment? PMID- 12076283 TI - Is it time to standardize granulocyte alloantigen nomenclature? PMID- 12076284 TI - The risk of transfusion-acquired CMV infection in seronegative solid-organ transplant recipients receiving non-WBC-reduced blood components not screened for CMV antibody (1984 to 1996): experience at a single Canadian center. AB - BACKGROUND: The use of CMV-safe cellular blood components has been recommended for CMV-sero- negative recipients of CMV seronegative (R-D-) solid- organ transplants. STUDY DESIGN AND METHODS: The incidence of CMV infection in 281 CMV seronegative patients receiving renal, heart, heart-lung, lung, and liver transplants at our center between January 1984 and October 1996 was studied. The blood components that these patients received were neither WBC reduced nor screened for CMV antibody. RESULTS: One hundred thirty-one of 154 (85.1%) patients receiving organs from CMV-seropositive donors (R-D+) developed CMV infection compared with three cases of presumptive transfusion-acquired (TA) CMV infection in 127 R-D- recipients (2.4%) (relative risk, 36.0; 95% CI, 11.8 110.4). The organ-specific incidence of TA CMV infection in R-D- patients was as follows: renal, 0 of 57 (0%); heart, 0 of 29 (0%); heart-lung and/or lung, 1 of 6 (16.7%); and liver, 2 of 20 (10%). There was no significant difference in the transfusion requirements of CMV-infected and -uninfected R-D- patients. False positive results were often (40%) observed when posttransfusion serum samples were used for determination of the organ donor CMV serostatus. CONCLUSION: The low risk of TA CMV infection observed in transplant patients who received standard blood components in our study should be considered when evaluating the efficacy of programs that provide CMV-safe blood components for this population. PMID- 12076285 TI - Quantitation of CMV by real-time PCR in transfusable RBC units. AB - BACKGROUND: CMV is one of the most significant pathogens infecting immunocompromised individuals. CMV is transmissible through transfusion of blood components. The goal of this study was to measure CMV levels in RBC units using a sensitive and quantitative DNA amplification assay. STUDY DESIGN AND METHODS: An assay to measure CMV load was developed by using real-time PCR to target the major immediate early viral gene. A probe (TaqMan, Applied Biosystems) was used to confirm product specificity and to permit quantitation of CMV in blood samples on a sequence detection system (ABI Prism 7700, Applied Biosystems). RESULTS: The assay was shown to be accurate, linear, and sensitive to as few as five copies of CMV DNA per PCR. The assay was applied to aliquots of RBC units from 203 healthy donors, 110 of whom were seropositive for CMV. CMV DNA was not detected in any of the 203 RBC samples. CONCLUSION: The findings statistically imply that at least 98.5 percent of RBC units have a CMV load of less than 250 copies per mL. Future clinical studies on larger numbers of units are required to determine the utility of real-time PCR in evaluating the risk of CMV transmission and in confirming the efficacy of WBC reduction. PMID- 12076286 TI - Stabilized viral nucleic acids in plasma as an alternative shipping method for NAT. AB - BACKGROUND: Preservation of the integrity of viral nucleic acids in blood specimens during shipping and handling is crucial for NAT and viral load monitoring. An economical and convenient method is described for nucleic acid stabilization by using an RNA stabilizing solution (RNAlater, Ambion) in plasma that is designed for the shipment of samples to tropical countries. STUDY DESIGN AND METHODS: HCV, HIV, and HBV FFP were compared with RNAlater-treated plasma and dried plasma spots (DPSs) after incubation at 37 degrees C, which was chosen as an upper limit of ambient shipping temperature, for up to 28 days. HCV-infected chimpanzee plasma was shipped at either room temperature after RNAlater treatment or as frozen plasma in liquid nitrogen from Liberia to New York City. They were then compared for HCV RNA levels. The nucleic acid stabilities were determined by quantitative PCR by using a molecular beacon assay on a sequence detection system (ABI 7700, PE-Biosystems) and by visualizing the PCR components on an acrylamide gel. RESULTS: Quantitative PCR data showed that a 60:40 or greater ratio of RNAlater:plasma volume successfully stabilized HCV RNA and HIV RNA in plasma for up to 28 days at 37 degrees C. HBV DNA in plasma was stable for up to 14 days at 37 degrees C without any stabilizing solution. DPSs on filter paper stabilized viral nucleic acids, but the recoveries were 3 to 10 times less than those with frozen plasma. The integrity of the 5' UTR region of HCV RNA in RNA later-treated chimpanzee plasma was intact when its PCR component was viewed on an acrylamide gel. CONCLUSION: The DPS method stabilized nucleic acids, at least with the extraction method used, was less sensitive than use of RNAlater, and required tedious manual handling. RNAlater provides a convenient way of stabilizing viral nucleic acid in plasma at ambient temperature during sample transportation. PMID- 12076287 TI - HIV seroconverting donors delay their return: screening test implications. AB - BACKGROUND: The yield of HIV p24 antigen testing implemented in March 1996 has been lower than projected. One possible explanation is that HIV seroconverting donors delay their return because of the recent practice of risk behaviors and/or signs and symptoms associated with primary infection. STUDY DESIGN AND METHODS: From a database of 6.8-million allogeneic donations collected at five U.S. blood centers from 1991 to 1997, 49 HIV, 21 HCV, 32 HTLV, and 44 HBsAg seroconverters with at least three donations were identified. A statistical method was developed to investigate whether the time between a donor's last negative donation and their positive donation was significantly longer than expected based on their previous return history. RESULTS: HIV seroconverters returned on average 42 percent later than expected (p < 0.01). Although not significant, HCV seroconverters donated on average 43 percent earlier than expected. HTLV and HBsAg seroconverters did not appear to change their donation pattern around the time of seroconversion. Sixty-three percent of the HIV seroconverters later acknowledged practicing a high-risk behavior. CONCLUSIONS: HIV seroconverters delay their return around the time of seroconversion and are thus less likely to be recently infected. Unique among HIV seroconverters, this observation provides a possible explanation for the lower than expected yield of HIV p24 antigen testing and suggests that NAT may have a similar low yield. PMID- 12076288 TI - Reduction of Yersinia enterocolitica load in deliberately inoculated blood: the effects of blood prestorage temperature and WBC filtration. AB - BACKGROUND: Yersinia enterocolitica is known to cause severe infections in patients who receive transfusions. STUDY DESIGN AND METHODS: The aim of the study was to define the best strategy for reducing the bacterial load in blood that was deliberately contaminated with Y. enterocolitica by combining prestorage temperature and WBC filtration with conditions of blood processing close to those applied in blood banks. RESULTS: The effects of three prestorage temperatures (4 degrees C, 20 degrees C, 37 degrees C) were evaluated at various times after infection. The best reduction of bacterial load was achieved after 3 hours at 20 degrees C. In further experiments, conducted according to the former specifications, filtration of whole blood from eight and six donors with an inoculum of 100 and 500 to 1000 CFUs per mL, respectively, resulted in a total inhibition of bacterial growth up to 42 days after infection. After fractionation of blood components, in contrast to plasma and RBCs, filtration was shown to reduce dramatically the bacterial growth in buffy coats, demonstrating that the antibacterial effect of filtration was supported by the removal of infected WBCs from blood samples. CONCLUSION: These results provide support for the systematic use of blood filtration in the preparation of blood components to prevent Y. enterocolitica infection of patients receiving transfusions. PMID- 12076289 TI - Collection of autologous monocytes for dendritic cell vaccination therapy in metastatic melanoma patients. AB - BACKGROUND: Dendritic cells (DCs) for immunotherapy of malignant melanoma can be generated from partially enriched monocytes prepared from PBMNCs. The feasibility of a single steady-state leukapheresis procedure to enrich monocytes for a complete vaccination series with up to 10 vaccinations was investigated. STUDY DESIGN AND METHODS: Thirty-eight patients (27 males and 11 females) with metastatic melanoma were enrolled in the study. All leukapheresis procedures were performed by a continuous flow method (Spectra, Cobe BCT) with a standard MNC program. RESULTS: An average of 11.7 L (range, 8-14 L) of whole blood was processed within 197.3 +/- 23.7 minutes, and a mean of 13.5 +/- 5.7 x 109 WBCs in a final volume of 191.0 +/- 24.2 mL was collected. The MNC purity in the apheresis component was 81.5 +/- 15.1 percent, from which 29.8 +/- 14.7 percent were monocytes. Thus, 11.0 +/- 5.0 x 109 MNCs and 3.2 +/- 2.0 x 109 monocytes were collected per procedure. Linear regression analysis revealed a high correlation between the absolute number of monocytes in peripheral blood before the apheresis procedure and the number of monocytes in the collected component (r=0.74, p < 0.0001). For the generation of DCs, 1.6 +/- 0.8 x 109 MNCs were plated into culture dishes; 3.2 +/- 1.8 percent of the cultured cells matured to DCs, which resulted in 56.5 +/- 49.4 x 106 DCs (range, 6.3-178) per patient for the complete vaccination series. CONCLUSION: A target dose of monocytes for the complete vaccination series could be obtained by a single convenient, safe, steady-state leukapheresis procedure in each patient without the need for G-CSF mobilization. The absolute number of monocytes in peripheral blood before the apheresis procedure is the best predictive variable for the yield of monocytes in the apheresis component. PMID- 12076290 TI - In vitro safety profile of G-CSF-mobilized whole blood after storage for 7 days in an infusable-grade L15 medium. AB - BACKGROUND: G-CSF-mobilized whole blood (WB) is a cost-reducing and simple alternative for peripheral blood progenitor cell transplantation. Recently, it was demonstrated that mobilized WB supplemented with Leibovitz's L15 medium permitted prolonged preservation of clonogenic cells at ambient temperature. In this study, an infusable-grade L15 medium (IG-L15) was developed, and the safety profile of mobilized WB after 7 days of storage was investigated. STUDY DESIGN AND METHODS: IG-L15 was manufactured in a closed system under good manufacturing practice conditions. Proinflammatory cytokine levels and hemolysis in mobilized WB were determined after 7 days of storage in different containers and were compared with current clinical mobilized WB values after 1 to 3 days of storage at 4 degrees C. RESULTS: IG-L15 and L15 maintained clonogenic cells equally. In the samples of mobilized WB that were returned to the patient, cytokine levels were not elevated in comparison with freshly collected mobilized WB. By using IG L15 in polystyrene-coated cell culture bags, median (range) levels of 9.4 (2.2 69.8) pg per mL (IL-1beta), 31.6 (6.1-146.5) pg per mL (TNF-alpha), 76.9 (15.5 934.9) pg per mL (IL-6), and 7195 (104-205,600) pg per mL (IL-8) were found after 7 days. Higher cytokine levels were found with L15 and different containers. He- molysis was less than 0.5 g per dL in all cases. CONCLUSION: The storage of mobilized WB for 7 days in IG-L15 at ambient temperature is possible with adequate preservation of clonogenic cells, but cytokine levels may require plasma removal before return. PMID- 12076291 TI - High-dose CD34+ cells are not clinically relevant in reducing cytopenia and blood component consumption following myeloablative therapy and peripheral blood progenitor cell transplantation as compared with standard dose. AB - BACKGROUND: No agreement exists about the number of autologous peripheral blood progenitor cells (PBPCs) to transfuse for optimal hematologic recovery after high dose chemotherapy. STUDY DESIGN AND METHODS: To determine CD34+ cell dosage following high-dose chemotherapy (in terms of hematologic recovery and blood component consumption), the effects of two schedules of CD34+ cell transfusions in a cohort of patients with myeloma or non-Hodgkin's lymphoma were examined. Forty patients (Group 1) received between 2.5 and 5 x 106 CD34+ cells per kg, with a median of 3.4 x 106 per kg following high-dose chemotherapy, and 40 patients (Group 2), selected to match Group 1 for age, diagnosis, prior therapies, and procedure for PBPC mobilization, received a dose of CD34+ cells >5 x 106 per kg, with a median of 8.4 x 106 per kg (5-33). RESULTS: The median number of days to achieve a neutrophil count of >0.5 x 109 per L and unsupported platelets of >20 x 109 per L was identical for the two groups, but the time required to reach 1.5 x 109 neutrophils per L and 50 x 109 platelets per L was greatly delayed in Group 1. No significant difference was observed for the median number of RBC and platelet transfusions, or for the proportion of patients in each group that did not require either platelet or RBC transfusions. CONCLUSION: Our data confirm a dose-response relationship between CD34+ cell dose transfused and time to hematologic recovery after high-dose chemotherapy. However, the minimal Hb and platelet counts for transfusion independence in the two groups are similar when the CD34+ cell dose is greater than 5 x 106 CD34+ cells per kg. Therefore, our data suggest that it is not necessary to go on with apheresis procedures after 5 x 106 CD34+ cells per kg are harvested to sustain one high dose chemotherapy. PMID- 12076292 TI - Intraoperative blood salvage: a mathematical perspective. AB - BACKGROUND: Intraoperative blood salvage (IBS) is one of several blood conservation strategies employed to limit the use of allogeneic blood. IBS with mathematical modeling was studied to gain a better understanding of its benefits and limitations. STUDY DESIGN AND METHODS: Equations, based on a convergent geometric series, allowed the assessment of maximum allowable blood losses to avoid allogeneic RBC exposure in relationship to the fractional recovery of IBS. Such equations allowed us to calculate the potential "savings" in allogeneic RBC volume and to define better the efficiency of IBS. RESULTS: This model predicts that substantial amounts of RBCs (>2 units) can be recovered if there is large blood loss (e.g., >3000 mL) and if the minimal allowable Hct is low (e.g., 21%). As with other blood- conservation methods, this article shows that there are limits to the potential benefit of IBS for the avoidance of allogeneic RBC transfusion. CONCLUSION: IBS can be modeled with a convergent geometric series, and the amount of blood loss is dependent on the difference between the patient's starting Hct, the minimum allowable Hct, and the fractional recovery of RBCs. Such modeling illustrates the limited utility of IBS as a sole blood-conservation strategy. PMID- 12076293 TI - The transfusion trigger and number of units transfused in patients with HIV: associations with disease stage and functional status. AB - BACKGROUND: The influence of quality of life (QOL), physical functioning, and HIV disease stage on the transfusion trigger and the number of units transfused was investigated. STUDY DESIGN AND METHODS: The Viral Activation Transfusion Study, a randomized, double-blind study at 11 participating sites, enrolled HIV-positive patients with anemia who required RBC transfusion; 428 patients were included in the analysis of the first transfusion. The QOL scores, Perceived Health Index, Karnofsky score, CD4+ cell count, HIV viral load, and site were analyzed for relationships with the Hb level and the number of units transfused. RESULTS: The transfusion trigger was lower in patients with higher levels of Karnofsky score, Perceived Health Index, CD4+ cell count, and a number of QOL scales. Both the Hb trigger and the number of units transfused had a significant site variation. Males were transfused at a significantly lower Hb level than females. In multivariate analysis, the CD4+ cell count remained significant, but the Karnofsky score or the Perceived Health Index did not. The number of RBC units transfused was associated with the Hb level, CD4+ cell counts, and Karnofsky scores in unadjusted analysis but with only Hb in adjusted analysis. CONCLUSIONS: In this group of HIV+ patients, lower CD4+ cell counts prompted transfusion at higher Hb levels. However, after controlling for the Hb level, the number of units transfused was associated with only the Hb level. The HIV stage appears to influence the decision to transfuse at a particular Hb level but not to influence the number of RBC units transfused. The functional status does not appear to influence the decision to transfuse. PMID- 12076294 TI - Report on the Fourth International Granulocyte Immunology Workshop: progress toward quality assessment. AB - BACKGROUND: A formal quality assurance (QA) scheme has been established to facilitate proficiency testing for granulocyte antibodies and antigens. STUDY DESIGN AND METHODS: Fifteen laboratories participated in the Fourth International Granulocyte Immunology Workshop. The main objective of the workshop was to establish a formal QA scheme for granulocyte serology and molecular typing methods. A secondary objective was to determine the relative sensitivities of the granulocyte immunofluorescence test, granulocyte agglutination test, and MoAb immobilization assays using defined antisera and protocols. RESULTS: Laboratories scored between 16.7 and 100 percent (mean, 57.5%) of the maximum available in the serologic part of this QA exercise. There were particular problems in detecting granulocyte-specific human neutrophil antigen-1 (HNA-1a) IgM antibodies and HNA 2a antibodies in the presence of HNA-1b antibodies. The granulocyte immunofluorescence test was more sensitive than the granulocyte agglutination test in titration studies, but the latter method more readily identified the presence of HNA-3a antibodies. HNA genotyping was generally well performed, with nine laboratories obtaining 100-percent correct results for HNA-1a, HNA-1b, and HNA-1c. CONCLUSIONS: There is a need to standardize the detection of granulocyte specific antibodies. Laboratories with good performance tended to use two methods for detecting granulocyte-specific antibodies and an HNA-typed panel of granulocytes. The use of a method for elucidating mixtures of granulocyte- and lymphocyte-reactive antibodies (e.g., MoAb immobilization assay) and the use of methods for detecting both cytotoxic and noncytotoxic HLA class I antibodies were also associated with a higher than average performance. PMID- 12076295 TI - FCGR3 variants and expression of human neutrophil antigen-1a, -1b, and -1c in the populations of northern Germany and Uganda. AB - BACKGROUND: Human neutrophil antigen-1c (HNA-1c) (SH) has been described as the third alloantigen of the Fc receptor type IIIb (FcgammaRIIIb) for IgG beside the known alloantigens HNA-1a (NA1) and HNA-1b (NA2). Controversy exists on the assignment of the HNA-1c coding gene to the FCGR3B locus and on a possible linkage between the HNA-1c and HNA-1a coding genes. STUDY DESIGN AND METHODS: Two hundred sixty northern German blood donors and 43 individuals from Uganda were typed for FCGR3B*1 (NA1), FCGR3B*2 (NA2), and FCGR3B*3 (SH) by allele-specific PCR. In a subset of FCGR3B*3-positive probands, PCR-amplified FCGR3 fragments were subcloned and sequenced. Transmission of FCGR3B*3 was analyzed in family studies. A possible correlation with the FcgammaRIIIb alloantigen expression was investigated by flow cytometry. RESULTS: In the northern German population, FCGR3B*3 was found exclusively in individuals carrying FCGR3B*1 independent of the existence of FCGR3B*2 at a frequency of 5 percent. In the individuals from Uganda, each possible combination of FCGR3B*1, FCGR3B*2, and FCGR3B*3 was detected. FCGR3B*3 frequency was 34.9 percent. Within both populations, some individuals carried each of the three genotypes. DNA sequencing revealed new FCGR3 variants caused by single nucleotide exchanges at the typical polymorphic positions. In one individual, six different FCGR3 variants were detected. CONCLUSION: The coincidence of the three known FCGR3B alleles varies within the population of Germany and Uganda. Three simultaneous FCGR3B forms may be explained by two gene loci, but the basis of the high number of different variants in some individuals still remains unclear. Possible explanations may be a hypermutation mechanism or a number of FCGR3 higher than expected hitherto. PMID- 12076296 TI - Use of enzyme treatment to enhance reactivity of HLA and platelet-specific antibodies in solid-phase RBC adherence assays. AB - BACKGROUND: Solid-phase RBC adherence (SPRCA) assays (Immucor) detect HLA and/or platelet-specific antibodies. Pretreatment of reagent platelets with enzymes was investigated to determine whether the sensitivity of the assay could be increased. STUDY DESIGN AND METHODS: SPRCA testing, following the manufacturer's directions, was performed on 51 sera from patients with either a positive SPRCA antibody screen, suboptimal clinical responses to platelet transfusions, and/or suspected immune thrombocytopenic purpura; testing was also performed following pretreatment of the reagent platelets with bromelin, papain, or ficin. Sera from 23 patients having negative routine SPRCA antibody screens and good clinical responses to transfusion were tested as controls. Lymphocytotoxic antibody testing was also performed on selected samples. The effectiveness of enzyme treatment was judged by the increase in the proportion of reagent platelets reacting with the sample and the observed reaction strengths. RESULTS: Pretreatments of reagent platelets with all three enzymes increased the reactivity of known antibodies and detected some HLA and platelet-specific antibodies that had not reacted in routine testing. The clinical significance of the antibody specificities detected only after enzyme pretreatment was verified by a correlation with results from transfusing antigen-negative units. Only occasional false-positive results after enzyme pretreatment were observed. CONCLUSIONS: The use of enzyme pretreatment of SPRCA screening strips can provide information that is useful in selecting appropriate units for transfusion. PMID- 12076297 TI - Isolation, characterization, and family study of DTI, a novel partial D phenotype affecting the fourth external loop of D polypeptides. AB - BACKGROUND: The Rh system is the most polymorphic of the blood group systems and is of major importance in transfusion medicine. The partial D phenotypes lack one or more of the D epitopes. These variants appear to have arisen through hybrid RhD-CE-D or by spontaneous point mutations in RhD. The serologic findings and the molecular characterization of a novel partial D phenotype, termed DTI, are presented here. STUDY DESIGN AND METHODS: RBCs from the DTI proband and RBCs from individuals with other partial D phenotypes were tested with MoAbs against 16 D epi- topes, according to the recommendations of the 4th ISBT Workshop on MoAbs (Rh Section 1A). A full-length cDNA encoding DTI and introns 4 and 5 of RhD were isolated and analyzed by DNA sequencing. A family study of the DTI allele was carried out using PCR-RFLP and long-range PCR methods. RESULTS: Analysis of RBCs from the proband revealed that the DTI phenotype lacks epitopes D1, D2.1 (partial), D2.2, D5, D6 (partial), and D8. The DTI polypeptide exhibits seven amino acid substitutions in the D polypeptide: F223V, A226P, E233Q, V238M, V245L, G263R, and K267M. The genomic organization of DTI showed that the replacement of RhD with RhCE was located in intron 4, and the replacement of RhCE with RhD was located in intron 5. Family studies revealed that the DTI allele was inherited maternally, whereas the RhD- allele was inherited paternally in the proband. CONCLUSION: The serologic data provide the first molecular characterization of DTI, a previously unknown partial D phenotype. This phenotype affected the D polypeptide within the fourth external loop, resulting in a new RhD-CE (entire exon 5)-D hybrid gene. It is worth noting that P226, encoded by exon 5, is derived from E of RhCE in the DTI polypeptide. Family studies demonstrated that DTI was associated with a cDTIE haplotype. PMID- 12076298 TI - Russia's National Research Center for Hematology: its role in the development of blood banking. PMID- 12076299 TI - Allogeneic hematopoietic stem cell transplantation and the risk for transmission of heritable malignancy. PMID- 12076300 TI - The first case of HCV infection detected before seroconversion in blood donors tested by HCV core antigen ELISA. PMID- 12076301 TI - Single-donor platelets reduce the risk of septic platelet transfusion reactions. PMID- 12076303 TI - IgE-mediated asthma and rhinitis I: a role of allergen exposure? AB - Exposures to airborne protein antigens, aeroallergens, may cause sensitization with production of Th2-dependent antibodies, including IgE. The IgE antibodies and associated cellular responses are responsible for the allergic airway diseases, allergic rhinitis and allergic asthma, which are increasing in societies with Western life style. Aeroallergens may have different potential to sensitize exposed subjects. Thus, there are only a limited number of important groups of aeroallergens, which are those from house dust mites, cockroaches, pets, pollens, and moulds. Allergy follows to a certain extent the pharmacological/toxicological paradigm of dose-response relationship. Unlike effects of pharmacologically and toxicologically active substances, allergens elicit their adverse effects in a two-stage process. In the first stage the immunologically naive individual is sensitized to the allergen. In the second stage renewed exposure to the allergen elicits the disease response. Also, high concentrations of aeroallergens may induce immunological tolerance. The scientific literature suggests that many environmental factors contribute to the increase in sensitization and development of airway allergies. Nevertheless, the dose-response relationships apply (within certain limits) both to the sensitization itself and to the exacerbation of the diseases. This suggest that exposure reduction may be one of the methods for reduction of risk, in relation to control of the allergic airway diseases. PMID- 12076304 TI - Bayesian pharmacokinetics of lithium after an acute self-intoxication and subsequent haemodialysis: a case report. AB - We report a case of a 39-year-old male with bipolar affective disorder who was admitted to hospital with an intentional acute lithium intoxication resulting in renal insufficiency. The patient had previously been treated with lithium, risperidone, fluoxetine and lorazepam, and successfully titrated to lithium levels of 0.7 mmol/l. After overdosing, the lithium level was 5.89 mmol/l and haemodialysis was initiated. A full pharmacokinetic time profile of lithium was obtained. After successful haemodialysis treatment, lithium levels recovered below toxic levels of 1.5 mmol/l in 53 hr. Without intervention non-toxic levels were not expected to have been reached within 6 days, based on computer simulation of predialysis levels. The patient was discharged 6 days after admission without residual symptoms. It was concluded that the lithium intoxication resulted from a combination of lithium overdose and subsequent renal insufficiency due to the overdose. A possible fluoxetine-risperidone interaction was not considered clinically apparent. PMID- 12076305 TI - Drug-induced ventricular tachyarrhythmia in isolated rabbit hearts with atrioventricular block. AB - The aims of this study were to develop a suitable model to study proarrhythmic potential using isolated rabbit hearts with atrioventricular block and to examine the proarrhythmic potential of several drugs using this model. With a normal K/Mg solution (K+=5.7 mM and Mg2+=1 mM), d,l-sotalol (10 and 30 microM), a class III antiarrhythmic drug, prolonged ventricular repolarization, such as QT intervals and monophasic action potential duration, and induced early after-depolarization and polymorphic ventricular tachyarrhythmia. Cisapride (0.1 and 0.3 microM), a 5 HT4 receptor agonist, also prolonged the ventricular repolarization, and induced early after-depolarization. With a low K/Mg solution (K+=1.5 mM and Mg2+=0.35 mM), d,l-sotalol at 30 microM and cisapride at 0.3 microM more potently prolonged the ventricular repolarization than with a normal K/Mg solution. Furthermore, the incidence of polymorphic ventricular tachyarrhythmia caused by cisapride at 0.3 microM with a low K/Mg solution was higher than that with a normal K/Mg solution. Mosapride citrate, another 5-HT4 receptor agonist, at 10 microM prolonged the ventricular repolarization and induced early after-depolarization with a low K/Mg solution, whereas the drug at 1 and 3 microM did not affect any of the parameters examined. Des-4-fluorobenzyl-mosapride, a metabolite of mosapride citrate, at 10 microM slightly prolonged the ventricular repolarization without inducing early after-depolarization or ventricular tachyarrhythmia. These results suggest that mosapride citrate and des-4-fluorobenzyl-mosapride have much less proarrhythmic potential than cisapride and that isolated rabbit heart with atrioventricular block, perfused with a low K/Mg solution, is a suitable model for predicting the proarrhythmic potential of drugs. PMID- 12076306 TI - Antioxidant/restorative effects of calcined gold preparations used in Indian systems of medicine against global and focal models of ischaemia. AB - Stroke, or ischaemic brain damage, is of great geriatric importance being the third most common cause of death after cancer and heart diseases in developed countries. Despite such high frequency, its management has received inadequate attention. Many studies have shown the role of free radicals in the pathogenesis of ischaemic brain damage. Search for safe and effective antioxidant and free radial scavenger agents, therefore, appear to be a promising approach for stroke therapy. Gold, widely used in modern medicine for the treatment of rheumatoid arthritis, is highly valued for various medicinal uses in Indian systems of medicine. Traditional gold preparations are attributed with tonic/rejuvenating and antioxidant properties. Our earlier studies revealed interesting analgesic, immunostimulant, adaptogenic and glycogen sparing properties in these preparations, but their effects in cerebral ischaemia have not been investigated. This prompted us to initiate the present study using global and focal models of ischaemia in albino rats. Enzymatic parameters (lipid peroxidase, reduced glutathione, catalase, glutathione reductase, glutathione-S-transferase, glutatione peroxidase, superoxide dismutase, and glucose-6-phosphate dehydrogenase) were employed to assess ischaemic brain damage and its modulation. Significant restoration of altered values to near normal levels by Ayurvedic Swarna Bhasma and Unani Kushta Tila Kalan (25 mg/kg, orally for 10 days), suggest potentials for gold preparations in cerebrovascular diseases. The preparations deserve more scientific attention for possible therapeutic exploitation. PMID- 12076307 TI - Tolterodine and its active 5-hydroxymethyl metabolite: pure muscarinic receptor antagonists. AB - Tolterodine and its major active 5-hydroxymethyl metabolite (5-HM) are potent muscarinic receptor antagonists that show selectivity for the urinary bladder over salivary glands in vivo. This tissue selectivity cannot be attributed to muscarinic receptor subtype selectivity, since both compounds are non-selective with respect to the M1-M5 receptor subtypes. The aim of the present in vitro study was to determine the specificity of tolterodine and 5-HM for muscarinic receptors compared to other potential cellular targets. Carbachol-induced contractions of isolated guinea pig bladder were effectively inhibited by tolterodine (IC50 14 nM) and 5-HM (IC50 5.7 nM). Tolterodine and 5-HM were weak inhibitors of effects mediated by alpha-adrenergic receptors (rat portal vein), histamine receptors (guinea pig ileum) and calcium channels (guinea pig potassium depolarised urinary bladder, spontaneously beating right atria and electrically driven right papillary muscle). The IC50 values were in the microM range and the antimuscarinic potency of tolterodine was 27, 200 and 370-485 times higher, respectively, than its potency in blocking histamine receptors, alpha adrenoceptors and calcium channels. The active metabolite, 5-HM, was >900 times less potent at these sites than at bladder muscarinic receptors. Radioligand binding data on 54 different receptors and binding sites showed that tolterodine and 5-HM bind with significant affinity only at muscarinic receptors. In conclusion, the results of the present study indicate that both tolterodine and 5 HM are specific muscarinic receptor antagonists. The tissue selectivity of these agents in vivo cannot therefore be explained by secondary pharmacological actions. PMID- 12076308 TI - Pharmacological antagonism of fumonisin B1 cytotoxicity in porcine renal epithelial cells (LLC-PK1): a model for reducing fumonisin-induced nephrotoxicity in vivo. AB - Fumonisin B1 is a mycotoxin commonly found on corn. It is hepatotoxic and nephrotoxic in domestic and experimental animals, and causes equine leukoencephalomalacia and porcine pulmonary oedema. It is a potent inhibitor of ceramide synthase. Inhibition leads to accumulation of free sphingoid bases in cells and tissues. In pig kidney epithelial cells (LLC-PK1), fumonisin B1 induces increased tumour necrosis factor alpha (TNFalpha) expression independent of the accumulation of sphingoid bases. The objective of this study was to investigate pharmacological approaches for intervening in fumonisin B1 toxicity using the LLC PK1 cell model. The toxicity of fumonisin B1 was assayed using cell viability and lactate dehydrogenase (lactate dehydrogenase) release. Pretreatment of cells with myriocin, preventing sphinganine accumulates, prevented the fumonisin B1-induced decrease in cell viability and increased lactate dehydrogenase release. Modulation of adenosine receptor activity did not reduce the fumonisin B1 cytotoxicity. As with myriocin, silymarin pretreatment prevented the fumonisin B1 induced effects on cell viability and lactate dehydrogenase release. When added 6 or 24 hr after treatment of cells with fumonisin B1, both myriocin and silymarin reversed the decreased cell viability and suppressed the increased lactate dehydrogenase release. Myriocin, but not silymarin, blocked the accumulation of sphinganine in fumonisin B1-treated cells. Silymarin, unlike myriocin, induced expression of TNFalpha to an extent similar to fumonisin B1, but pretreatment with silymarin decreased the fumonisin B1-induced TNFalpha expression in LLC-PK1 cells. Results suggest that the mechanisms by which myriocin and silymarin protect renal cells are different, and silymarin potentially prevents fumonisin B1-induced toxicity by modulating TNFalpha expression or signals downstream of the inhibition of ceramide synthase. PMID- 12076309 TI - Protective effect of aloe extract against the cytotoxicity of 1,4-naphthoquinone in isolated rat hepatocytes involves modulations in cellular thiol levels. AB - Aloe is a familiar ingredient in a wide range of health care and cosmetic products and has been reported to possess various physiological effects, antioxidative, anticarcinogenic, antiinflammatory and laxative. Aloe has also been reported to have an effect on liver function. The cytoprotective effect of aloe extract against 1,4-naphthoquinone-induced hepatotoxicity was evaluated in primary cultured rat hepatocytes. After exposure to 1,4-naphthoquinone (100 microM), a decrease in cell viability measured as >60% lactate dehydrogenase depletion was induced. Cellular glutathione (GSH) and protein-SH levels were also significantly decreased in a time-dependent manner. However addition of aloe extract resulted in a dose-dependent improvement of these effects. This cytoprotective effect of aloe could be attributed to its inhibition of GSH and protein-SH depletions. The effect of the aloe extracts were also dose-dependent. Addition of diethyl maleate (1 mM), a cellular glutathione-depleting agent, to hepatocytes treated with both 1,4-naphthoquinone and aloe extract, induced depletion of GSH, but did not affect protein-SH or lactate dehydrogenase. These results suggest that the 1,4-naphthoquinone-induced toxicity in rat hepatocytes was inhibited by aloe extract, and that this protective effect was due to the maintenance of cellular thiols, especially protein-SH. PMID- 12076310 TI - Treatment of distal colitis with local anaesthetic agents. AB - The results of clinical and experimental studies on topical treatment of distal colitis with local anaesthetic agents are summarized. The original observation was an adrenergic hyperinnervation of the inflamed mucosa (hyperinnervation hypothesis). In order to silence local nervous reflexes, the mucosa was treated topically with 2% lidocaine gel. The clinical results are promising and no side effects have been observed. The relapse rate is relatively high and related to the duration of treatment. In studies of experimental colitis a potential antagonism between harmful adrenergic nerves (vasoconstrictor substances and proinflammatory cytokines) and mucosa-protective visceral afferents (antiinflammatory cytokines) in the pathogenesis of colitis is intriguing. Other studies have emphasized the importance of neutrophils for causing damage to the colon epithelium (neutrophil hypothesis) and local anaesthetics have potent effects on several steps of the inflammatory response in addition to the nervous blockade. PMID- 12076311 TI - Functional impairment of renal organic cation transport in experimental diabetes. AB - This study was designed to determine the effect of diabetes on the function of the renal organic cation transport system that mediates the excretion of a wide variety of toxicants and drugs. The experiments compared the ability of renal cortex slices from streptozotocin-induced diabetic and non-diabetic rats to accumulate the model cation, 14C-tetraethylammonium under controlled conditions. Initial experiments demonstrated a progressive decline in tetraethylammonium accumulation with increasing duration of diabetes. The maximal decrease was observed at 21 days after streptozotocin injection. Time-dependent incubations revealed that tetraethylammonium uptake from both diabetic and non-diabetic rats followed a curvilinear pattern expected of an active process. However, at steady state the diabetic-derived slices accumulated a significant 38% less tetraethylammonium versus slices from non-diabetics. Concentration-dependent incubations of tetraethylammonium (0.01-10 mM, 60 min.) demonstrated saturable transport in both diabetic and non-diabetic slices with a significantly decreased capacity of diabetic-derived slices to accumulate tetraethylammonium. Cellular respiration rates in the two groups were not different. Insulin treatment of the diabetic rats prevented the transport decline. While the causative factor of the transport impairment in diabetes is unresolved, this study documents an aspect of diabetic nephropathy that has not been previously reported but which may have important implications for renal excretion of cationic drugs and toxicants. The results also provide a mechanism for the well-documented "protection phenomenon" by which the kidneys of diabetic rats are resistant to nephrotoxicity induced by the chemotherapeutic agent cisplatin. PMID- 12076312 TI - Intravenously administered oxotremorine and atropine, in doses known to affect pain threshold, affect the intraspinal release of acetylcholine in rats. AB - Both systemically and intrathecally administered cholinergic agonists produce antinociception while cholinergic antagonists decrease pain threshold. The mechanism and the site of action of these substances are not known. In the present study it was hypothesized that systemically administered muscarinic agonists and antagonists modify nociceptive threshold by affecting intraspinal release of acetylcholine (ACh). Catheters were inserted into the femoral vein in rats maintained on isoflurane anaesthesia for administration of oxotremorine (10 300 microg/kg) and atropine (0.1, 10, 5000 microg/kg). Spinal microdialysis probes were placed intraspinally at approximately the C2-C5 spinal level for sampling of acetylcholine and dialysis delivery of atropine (0.1, 1, 10 nM). Additionally, the tail-flick behaviour was tested on conscious rats injected intraperitoneally with saline, atropine (10, 100 and 5000 microg/kg), or subcutaneously with oxotremorine (30, 100, 300 microg/kg). Subcutaneous administration of oxotremorine (30, 100, 300 microg/kg) significantly increased the tail-flick latency. These doses of oxotremorine dose-dependently increased the intraspinal release of acetylcholine. Intravenously administered atropine, in a dose that produced hyperalgesia (5000 microg/kg) in the tail-flick test, significantly decreased the intraspinal release of acetylcholine. Our results suggest an association between pain threshold and acetylcholine release in spinal cord. It is also suggested that an approximately 30% increase in basal ACh release produces antinociception and that a 30% decrease in basal release produces hyperalgesia. PMID- 12076313 TI - Aloe emodin suppresses myofibroblastic differentiation of rat hepatic stellate cells in primary culture. AB - We have studied the inhibitory effect of aloe emodin on hepatic stellate cells activation and proliferation, as these cells play a key role in the pathogenesis of hepatic fibrosis. Rat hepatic stellate cells were activated by contact with plastic dishes, resulting in their transformation into myofibroblast-like cells. Primary hepatic stellate cells were exposed to aloe emodin (1-10 microg/ml). Possible cytotoxic effects were measured on stellate cells and hepatocytes using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of aloe emodin on production of type I collagen and smooth muscle cell alpha-actin were examined at the same concentration, by quantitative immunoprecipitation. Antiproliferative effects were examined by bromodeoxyuridine incorporation. Aloe emodin at 10 microg/ml restored the morphological changes characteristic of activated primary stellate cells, reduced DNA synthesis to 95% of control hepatic stellate cells at 10 microg/ml without affecting cell viability, and inhibited type I collagen production and smooth muscle alpha-actin expression by 86.77% and 99%, respectively, which suggest that aloe emodin is a potent inhibitor of stellate cell transformation. PMID- 12076314 TI - The mechanism underlying the protective effect of the thearubigin fraction of black tea (Camellia sinensis) extract against the neuromuscular blocking action of botulinum neurotoxins. AB - The aim of the present study was to elucidate the mechanism of the protective effect of black tea extract, the thearubigin fraction, against the neuromuscular blocking action of botulinum neurotoxin types A, B, and E. The effects of thearubigin fraction extracted from a black tea infusion were examined on the neuromuscular blocking action of botulinum neurotoxin types A, B, and E in mouse phrenic nerve-diaphragm preparations and on the binding of these toxins to rat cerebrocortical synaptosomes. Botulinum neurotoxin type A (1.5 nM), B (6 nM), or E (5 nM) abolished indirect twitches in mouse phrenic nerve-diaphragm preparations within 50, 90, 90 min., respectively. Thearubigin fraction mixed with each toxin blocked the inhibitory effect of the toxins. The specific binding of [125I]botulinum neurotoxin type A, B, or E to rat cerebrocortical synaptosomes was inhibited by mixing iodinated toxin with thearubigin fraction. The elution profile of [125I]botulinum neurotoxin type A, B, or E on Sephadex G-50 column chromatography was different from that of toxin mixed with thearubigin fraction. These findings indicate that thearubigin fraction protects against the neuromuscular blocking action of botulinum neurotoxin types A, B, and E by binding with the toxins. PMID- 12076315 TI - Age-related alteration in hepatic disposition of the neurotoxin 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine and pesticides. AB - Idiopathic Parkinson's disease may be caused by environmental neurotoxins such as pesticides, however the major risk factor is old age. We postulated that the high incidence of Parkinson's disease in older people is secondary to age-related impairment of the hepatic detoxification of xenobiotics. Previously, we have shown that there are significant differences between the hepatic disposition of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and pesticides. Here, we investigated whether there are age-related differences in the hepatic disposition of MPTP and pesticides, putatively associated with the pathogenesis of Parkinson's disease. We measured the hepatic disposition of paraquat, dichlorodiphenyltrichloroethane (DDT), malathion and MPTP using the multiple indicator dilution technique in the perfused livers of Fischer F344 rats aged 3 and 18 months. The recoveries of MPTP, DDT and malathion were increased from the livers of the older rats (by 258%, 253% and 134% compared with young rats, respectively). The hepatic transport of DDT and malathion into hepatocytes was reduced with age suggesting that part of the impaired uptake of neurotoxins may be secondary to an age-related barrier to influx. Ageing may increase risk of Parkinson's disease by altering hepatic detoxification and increasing systemic bioavailability of neurotoxins. PMID- 12076316 TI - Effects of carbamazepine and novel 10,11-dihydro-5H-dibenz[b,f]azepine-5 carboxamide derivatives on synaptic transmission in rat hippocampal slices. AB - The effects of carbamazepine on synaptic transmission in rat hippocampal slices were compared with those of two novel analogues (BIA2-093 and BIA2-024) with equivalent anticonvulsant efficacy but with fewer side effects. Carbamazepine (10 1000 microM) inhibited in a concentration-dependent manner the field excitatory postsynaptic potential (fPSP) response, with an EC50 of 263 microM, and also attenuated the presynaptic volley with a similar EC50 value. Carbamazepine was more potent to inhibit the NMDA receptor component of the fPSP (fPSPNMDA), with an EC50 of 160 microM. BIA2-093 and BIA2-024 were nearly equipotent with carbamazepine to inhibit synaptic transmission, and displayed similar potency to inhibit the fPSP (EC50 of 145 microM and 205 microM) and fPSPNMDA responses (EC50 of 198 microM and 206 microM). As with carbamazepine, BIA2-093 and BIA2-024 also attenuated the presynaptic volley with EC50 values ranging from 142 to 322 microM. These results indicate that carbamazepine and its analogues mostly inhibit synaptic transmission through inhibition of conduction, although carbamazepine, but not BIA2-093 and BIA2-024, may also depress NMDA receptor mediated responses. PMID- 12076317 TI - Delta-aminolevulinate dehydratase inhibition by phenyl selenoacetylene: effect of reaction with hydrogen peroxide. AB - The effect of phenyl selenoacetylene and its selenoxide on delta-aminolevulinate dehydratase from liver of adult rats (mammalian source) and from cucumber leaves (plant source) was investigated. In vivo, selenides can be oxidized to selenoxides by flavin-containing monooxygenases and selenoxides can regenerate selenides by thiol oxidation. The compound phenyl selenoacetylene was converted to selenoxides by reaction with hydrogen peroxide. Phenyl selenoacetylene inhibited mammalian and plant delta-aminolevulinate dehydratase with an IC50 about 250 microM and >400 microM, respectively. Its selenoxide inhibited the enzyme more strongly, with IC50 values of 45 microM and 100 microM for the mammalian and plant source, respectively. The selenoxide inhibitory action was antagonized by dithiothreitol suggesting the involvement of -SH groups. Moreover, delta-aminolevulinate dehydratase from a plant source was inhibited by the selenoxide, suggesting a possible involvement of -SH groups located at a site distinct from the region implicated in Zn2+ binding in mammalian delta aminolevulinate dehydratase. The results of the present study suggest that (i) delta-aminolevulinate dehydratase is a potential molecular target for phenyl selenoacetylene, due to the oxidation of enzyme sulfhydryl groups, and that (ii) the monooxygenation of this selenocompound, which in vivo could be possibly mediated by flavin-containing monooxigenases, increases its inhibitory effect. PMID- 12076318 TI - Bioavailability of reduced nicotinamide-adenine-dinucleotide (NADH) in the central nervous system of the anaesthetized rat measured by laser-induced fluorescence spectroscopy. AB - Drugs intended to increase wellness or quality of life ("lifestyle drugs") have gained popularity and/or importance over recent years. Biogenic substances like nicotinamide adenine dinucleotide (NADH) are supposed to increase the physical and intellectual performance without side-effects. NADH is an energy-delivering co-substrate in the respiratory chain. Clinical studies showed positive effects of peripherally given NADH in Morbus Parkinson and major depression. NADH can be measured by its fluorescence. In this study a pulsed N2-laser combined with a fibre-optic probe and photomultipliers was used to induce and measure NADH fluorescence in the rat cortex. The aims of the study were to assess the suitability of the laser-induced spectroscopy for in vivo and on-line measurement of NADH in neuroscience and the assessment of the central availability of NADH after peripheral administration. NADH (50 mg/kg) but not the precursor nicotinamide caused a significant rise of the NADH fluorescence intensity indicating an increase of the NADH concentration in the rat cortex. In conclusion, the results suggest that NADH given orally or intraperitoneally increases the amounts of NADH in the brain. The results may thus help to explain the clinical effects reported. PMID- 12076319 TI - Ethanol inhibits store-operated Ca2+ entry of platelets. PMID- 12076321 TI - Mast cell/T cell interactions in oral lichen planus. AB - Lichen planus is a disorder characterized by lesions of the skin and oral mucous membranes. Although many patients have involvement of both skin and oral mucosa at some stage during the progress of the disease, a larger group has oral involvement alone. It has been reported that oral lichen planus (OLP) affects one to two percent of the general population and has the potential for malignant transformation in some cases (1, 2). Like many chronic inflammatory skin diseases, it often persists for many years. Numerous disorders may be associated with OLP such as graft-vs.-host disease and Hepatitis C virus infection (3), however, it is unclear how such diverse influences elicit the disease and indeed whether they are identical to idiopathic OLP. Available evidence supports the view that OLP is a cell-mediated immunological response to an induced antigenic change in the mucosa (4-6). Studies of the immunopathogenesis of OLP aim to provide specific novel treatments as well as contributing to our understanding of other cell-mediated inflammatory diseases. In this paper, the interactions between mast cells and T cells are explored from the standpoint of immune regulation. From these data, a unifying hypothesis for the immunopathogenesis of OLP is then developed and presented. PMID- 12076322 TI - Serum interleukin-6 level is a useful marker in evaluating therapeutic effects of levamisole and Chinese medicinal herbs on patients with oral lichen planus. AB - BACKGROUND: Oral lichen planus (OLP) is a T cell-mediated inflammatory disease. Interleukin-6 (IL-6) is a pro-inflammatory cytokine that has effects on cellular and humoral immunities. Previous studies have shown that keratinocytes and tissue infiltrating mononuclear cells from OLP lesions can secrete IL-6. In some OLP patients, the high serum IL-6 levels are reduced after treatment, suggesting that IL-6 may be a useful marker in evaluating therapeutic effects and in monitoring the disease status of OLP. METHODS: In this study, we used a solid phase, two site sequential chemiluminescent immunometric assay to determine the baseline serum levels of IL-6 in a group of 180 patients with erosive OLP (EOLP), nonerosive OLP (NEOLP), erythema multiforme (EM), traumatic ulcers (TU), oral submucous fibrosis (OSF), pemphigus vulgaris (PV), or Sjogren's syndrome (SS), and in 77 normal control subjects. Some OLP patients were treated with levamisole plus Chinese medicinal herbs or levamisole only for 0.5-5.5 months and their serum IL-6 levels were measured after treatment. RESULTS: We found that approximately 99% of the normal control subjects and the patients with EM, TU, or OSF had a normal serum IL-6 level less than 5.0 pg/ml. However, 15% (22/149) OLP patients, 15% (20/136) EOLP patients, 20% (5/25) major type EOLP patients, 14% (15/111) minor type EOLP patients, 15% (2/13) NEOLP patients, 14% (1/7) EM patients, 43% (3/7) PV patients, and 100% (6/6) SS patients had a serum IL-6 level greater than 5.0 pg/ml. The mean serum IL-6 level in patients with OLP (3.4 +/- 3.1 pg/ml, P < 0.001), EOLP (3.4 +/- 3.2 pg/ml, P < 0.001), major type EOLP (4.9 +/- 3.5 pg/ml, P < 0.001), minor type EOLP (3.0 +/- 3.0 pg/ml, P < 0.01), or NEOLP (4.2 +/- 1.5 pg/ml, P < 0.001) was significantly higher than that in normal control subjects (2.0 +/- 1.5 pg/ml). A significant difference in the mean serum IL-6 level was also found between major type and minor type EOLP patients (P < 0.01). The mean reduction of serum IL-6 level in OLP patients treated with levamisole plus Chinese medicinal herbs was significantly higher (7.4 +/- 4.7 pg/ml) than that in OLP patients treated with levamisole only (3.8 +/- 2.3 pg/ml, P < 0.05), suggesting that the combination therapy was superior to levamisole only. CONCLUSION: We conclude that levamisole and levamisole plus Chinese medicinal herbs can modulate the serum IL-6 level in OLP patients. IL-6 may be a useful marker in evaluating therapeutic effects and in monitoring the disease status of OLP. PMID- 12076323 TI - Cell proliferation activity and the expression of cell cycle regulatory proteins in oral lichen planus. AB - BACKGROUND: In oral lichen planus (OLP), destruction of the basal cell layer, which is one of the characteristic histological features, is seen and many changes in cell proliferation, cell repair and cell death occur in the injured mucosal epithelium. METHODS: We studied mucosal tissues from 19 patients of OLP and 10 controls, with immunohistochemistry for Ki-67, p53, cyclin dependent kinase inhibitors (CDKI) and cyclins. Mitotic count was calculated. TUNEL assay was also performed for evaluation of apoptotic cell death. RESULTS: Mitotic count, Ki-67 and cyclin D1 labeling indices in the basal and parabasal layers of OLP mucosa were elevated in comparison with those of controls. p53, p21Cip1 and TUNEL indices of OLP mucosa were also increased. CONCLUSIONS: These complex changes, which concomitantly occur in the injured mucosal epithelium, could contribute to the development and maintenance of characteristic mucosal epithelial architectures seen in OLP. PMID- 12076324 TI - The incidence of oropharyngeal cancer in Taiwan: an endemic betel quid chewing area. AB - BACKGROUND: Oropharyngeal cancer is the one of the most common cancers in the world. The purpose of this study was to examine the trends in oropharyngeal cancer from 1979 to 1996 in Taiwan. METHODS: Traditional cohort analysis was employed to show the birth-cohort effect of oropharyngeal cancer incidence. Age period-cohort model analysis was used to examine the age, period and cohort effect between intraoral and pharyngeal cancer. RESULT: A significant increasing trend in oropharyngeal cancer has been seen in males. The principal increases have been seen in tongue and mouth in males, and tongue in females. In males, an increasing trend was found in successive cohorts born after 1929. The increase in incidence of intraoral cancer of males was greater than the increase in incidence of pharyngeal cancer in the younger age group, more recent time periods and the younger cohorts. CONCLUSION: The increasing incidence in oropharyngeal cancer in Taiwan is probably heavily influenced by the rising consumption of alcohol and use of betel quid. The effect of these etiologies on intraoral cancer was more significant in recent time periods, later cohorts and the younger group. PMID- 12076325 TI - The retinoic acid receptor-beta (RAR-beta) mRNA expression in the oral squamous cell carcinoma associated with betel quid use. AB - BACKGROUND: Within the abundant retinoids nuclear receptors, abnormally low expression of the RAR-beta has been shown to contribute to neoplastic progression in oral epithelium in western countries. Distinctly different risk factors contributing to oral squamous cell carcinoma (OSCC) in epidemiologically different societies denote the value of exploring the role of RAR-beta expression in OSCC associated with betel quid (BQ) use in our society. METHODS: We examined the cellular expression of RAR-beta using in situ hybridization (ISH) analysis on 38 pairs of surgical specimens of primary OSCC and non-cancerous matched tissues (NCMT) to correlate with their clinico-pathological features including age, sites of tumor, habit of BQ use, stage, size of primary tumor, lymph node metastasis, differentiation. RESULTS: Of all cases analyzed, BQ users were significantly younger than non-BQ users (51.2 +/- 2.1 vs. 60.2 +/- 2.6, P = 0.01). 52% OSCC of BQ users (13/25) and 23% OSCC of non-BQ users (3/13) exhibited the absence of RAR beta expression. In 17 paired-samples from buccal mucosa (BM), most NCMT and less than half of OSCC exhibited RAR-beta expression (16/17, 94% vs. 8/17, 47%, P = 0.003). The RAR-beta expression was seen in the vast majority of the well differentiated OSCC and in less than half of the moderately differentiated OSCC only (15/20, 75% vs. 7/18, 39%, P = 0.03). CONCLUSION: A correlation between the loss of RAR-beta expression and more advanced histopathological grade tumors was observed. This study also suggests that the loss of RAR-beta expression is significant in BM OSCC, which preferentially occurs in BQ users. PMID- 12076326 TI - Prediction of delayed neck metastasis in patients with stage I/II squamous cell carcinoma of the tongue. AB - BACKGROUND: The incidence of delayed neck metastasis (DNM) in patients with squamous cell carcinoma (SCC) of the tongue is reported to be 20% to 50%. Although clinically negative cervical lymph nodes (N0) are associated with a good outcome, the prognosis is poor in patients with DNM. The aim of this study was to evaluate the clinicopathological and immunohistochemical parameters associated with DNM in patients with stage I/II SCC. METHODS: Fifty nine patients, with previously untreated stage I/II carcinoma, underwent examination of clinicopathological and immunohistochemical parameters and incidence of DNM. A linear discriminant analysis was used to analyze prognostic factors and to determine the probability of DNM occurring. RESULTS: DNM occurred in 14 (24%) subjects of the 59 study patients, level I to level III, within 5 years. Parameters such as gender and age, disease stage, tumor size and histological grade, tumor location, degree of tumor invasion and expression of VEGF, E cadherin or Ki-67 showed no significant correlation with the occurrence of DNM; however, factors such as tumor morphology, tumor thickness greater than 4 mm, and Flt-4 expression were significantly associated with development of DNM. CONCLUSIONS: Such factors provide useful information with regard to DNM and the prognosis. We concluded that patients with early SCC whose tumors are > 4 mm in thickness and immunopositive for Flt-4 are particularly at risk of developing DNM. PMID- 12076327 TI - Expression of hMSH2 and hMLH1 proteins of the human DNA mismatch repair system in salivary gland tumors. AB - BACKGROUND: The human DNA mismatch repair (hMMR) system plays an important role in reducing mutation and maintaining genomic stability. The MMR system in human cells is composed of at least six genes (hMSH2, hMLH1, hMSH3, hPMS1, hPMS2 and GTBP/hMSH6). In particular, hMSH2 and hMLH1 are expressed in cells undergoing rapid renewal; their reduced expression has been reported in several tumors. METHODS: We examined the expression of hMSH2 and hMLH1 by immunohistochemistry in tumor specimens from 43 patients with primary tumors. RESULTS: All carcinomas (n = 20) expressed these proteins. In addition, when compared to pleomorphic adenomas, malignant tumors contained significantly (P < 0.01) higher proportions of hMSH2 (56.1 +/- 31.5 vs. 31.1 +/- 22.6) and hMLH1 (27.9 +/- 26.0 vs. 14.0 +/- 12.6) positive cells. Warthin's tumors showed no specific nuclear staining of tumor cells for both hMSH2 and hMLH1. CONCLUSIONS: These data suggest a minor, if any, role for a defect in the hMMR system in the pathogenesis of malignant salivary gland tumors. PMID- 12076328 TI - Clinico-pathological aspects of a residual natal tooth: a case report. AB - A Japanese girl was referred to Osaka University Dental Hospital for examination of a tooth-like structure that had erupted following spontaneous exfoliation of a natal tooth in the lower left primary central incisor region. The structure had erupted at 6 months of age, and radiographic and clinical examination showed composition of pulp and dentin, but no enamel. On histological examination, the majority of the dentin area had a tubular dentin-like appearance, while the outer area of the root appeared to be composed of an osteodentin-like substance. Most of the dentin was covered by cementum. These findings suggest that the structure had originated from a developing remnant of the extracted natal tooth, which must have remained in the gingival tissues. We termed this calcified structure a residual natal tooth. PMID- 12076329 TI - Sebaceous lymphadenoma of the lip: report of a case of minor salivary gland origin. AB - A case of sebaceous lymphadenoma occurring in the lip of a 73-year-old female is described. The patient had noticed a painless mass in the region of her upper lip for a year. The surgically removed tumor, measuring about 10 mm in diameter, was located just beneath the lip mucosa, expanding into the submucosal and muscle layer. Histologically, the tumor was well encapsulated and consisted of scattered round-shaped islands of small squamous epithelial cells with focal but apparent sebaceous differentiation in a background of lymphoid stroma. This is the first case report of sebaceous lymphadenoma of minor salivary gland origin. PMID- 12076330 TI - Metastatic infiltration of the dental pulp by medulloblastoma. AB - Medulloblastoma is an infratentorial primitive neuroectodermal tumour. It is the most commonly occurring brain tumour of childhood, accounting for 15-20% of all paediatric tumours. Extracranial metastasis is rare, but may involve the skeleton. Jaw lesions, however, have never been described. A case is reported of metastases of a medulloblastoma to the jaw including the dental pulp. PMID- 12076331 TI - The role of molecular techniques in preventing oral cancer and improving its poor prognosis: an illusion? PMID- 12076333 TI - Gli-type zinc finger proteins as bipotential transducers of Hedgehog signaling. AB - Secreted proteins of the Hedgehog (Hh) family direct the development of diverse organs and tissues of vertebrates and invertebrates. Gli-type zinc finger proteins function as transcriptional mediators of the Hh signaling cascade and were implicated both in the activation and repression of Hh target genes. The differential activity of Gli-type zinc finger proteins is regulated on the level of proteolytic processing and subcellular localization as a complex concert of Hh responsive, intracellular determinants. Here, we provide a survey of recent studies on the characterization of molecular mechanisms involved in the interpretation of Hh signals by Gli-type zinc finger proteins. PMID- 12076334 TI - Mucociliary differentiation according to time in human nasal epithelial cell culture. AB - Knowledge of the state of differentiation, cell phenotype, and expression of genes for mucus production at the time of study is important because these may vary at different times during the culture period. The primary purpose of this study was to determine whether the number of ciliated cells increases as a function of differentiation in NHNE cells. If we observed an increase in the number of ciliated cells, the composition ratio of ciliated and secretory cells according to the culture duration was determined. The levels of mucin and lysozyme secretion and their gene expression at this time were also examined. The presence of ciliated cells was not evident up to 2 days after confluence. However, 3.1 +/- 0.2 %, 7.4 +/- 0.5 %, and 14.5 +/- 0.6 % of the cells were ciliated on the 7th, the 14th, and the 28th day after confluence, respectively. Meanwhile, the percentage of secretory cells were 35.6 +/- 2.8 %, 32.8 +/- 2.5 %, 32.8 +/- 2.5 %, and 49.4 +/- 1.4 % on the 2nd, the 7th, 14th, and 28th day after confluence. The amount of secreted mucin showed an abruptly increasing pattern by the 14th day after confluence but showed no significant changes thereafter. The amount of secreted lysozyme increased as a function of differentiation. MUC5AC and MUC5B mRNA were mainly expressed between the 7th and the 14th day after confluence with relatively weak MUC8 and lysozyme expression. By the 28th day after confluence however, as the MUC5AC mRNA expression became weaker, MUC5B, MUC8, and lysozyme mRNA expression became stronger. In conclusion, we speculate that in in vitro studies with NHNE cells, the time point of treatment should vary according to the purpose of the study. In addition, the MUC5B and MUC8 gene may play an important role in mucin secretion in fully differentiated human nasal epithelial cells. PMID- 12076335 TI - Positive effect of collagen V and VI on triglyceride accumulation during differentiation in cultures of bovine intramuscular adipocytes. AB - Ethyl-3,4-dihydroxybenzoate (EDHB), a specific inhibitor of collagen synthesis, was used to study the role of collagen in the differentiation of bovine intramuscular preadipocytes (BIP). Triglyceride (TG) accumulation levels of BIP cells were dose-dependently inhibited by EDHB and were reduced to 50 % at a 0.1 mM concentration. EDHB addition prevented the accretion of collagens (types I-VI) on the cell surface, which generally increases during adipose conversion. Western blotting and immunofluorescence studies showed in detail that triple-helical conformation of procollagen molecules was drastically interrupted by EDHB, and as a result, their matrix assembly was not performed in the extracellular space of adipocytes. Particularly, the development of collagen types IV, V and VI during differentiation was severely damaged. When exogenous collagens were supplied to make up for the lack of endogenous products, cultured EDHB-treated cells on type V and VI collagen-coated dishes were the only ones among six collagens to accumulate more TG, although their TG content did not reach that of normal adipocytes. This result implies the importance and the active role of collagens V and VI for adipogenesis. However, these findings also indicate that collagen newly synthesized and organized by the adipocyte itself during differentiation is still necessary for the growth of adipose tissue. PMID- 12076336 TI - Expression of multiple troponin T isoforms in chicken breast muscle regeneration induced by sub-serous implantation. AB - Chicken fast-muscle type (F-type) troponin T (TnT) isoforms are classified into two types, leg-muscle type (L-type) and breast-muscle type (B-type), which are generated by exclusion and inclusion of exon x series-derived sequences in mRNAs, respectively. The B-type isoforms are further classified into neonatal breast muscle (BN), young chicken breast-muscle (BC), and adult chicken breast-muscle (BA) subtypes. It is known that the multiple F-type TnT isoforms are transiently expressed in the breast muscle tissue during normal development. To examine whether the transition of the isoforms was fixed in muscle cell lineage, breast muscle pieces (pectoralis major) of 1-day old chicks were cultured under gizzard serous membrane of the same chicks for 60 days at the longest. TnT isoform expression of the implants was monitored by immunoblotting and immunostaining using anti-F-type TnT against both L-type and B-type isoforms, anti-exon x3 against only B-type isoforms, and anti-S-type TnT against slow-muscle-type (S type) isoforms. Muscle fibers in the implant degenerated first, and then new myotubes expressing L-type isoforms were formed by the fusion of myoblasts from surviving satellite cells. When the maturation of the myotubes into myofibers proceeded, BN-, BC-, and BA-subtype isoforms were expressed in the order of developmental stage specific-manner, indicating that the order of appearance of these isoforms was fixed in muscle cell lineage. In immunostaining of the implants recovered on the 60th day after implantation, at least three kinds of the regenerated myofibers were observed, expressing mainly B-type, both B-type and L-type, and only L-type isoforms. The immunohistochemical results suggested that the regulation of alternative splicing of F-type TnT pre-mRNAs was different among individual myofibers, and that the regulation was programmed in myogenic cells, probably satellite cells, which were the primary source of the fibers. PMID- 12076337 TI - Regulated lens regeneration from isolated pigmented epithelial cells of newt iris in culture in response to FGF2/4. AB - When a lens is removed from the newt eye, a new lens is regenerated from the pigmented epithelial cells of the dorsal iris, whereas the ventral iris never shows such an ability. It is important to clarify the nature of signaling molecules which act directly on the iris cells to accomplish lens regeneration from the iris and also to gain insight into the mechanism of dorso-ventral difference of the regeneration potential. To examine the effects of exogenous factors, we established an in vitro culture of reaggregates made from dissociated pigmented epithelial cells of dorsal or ventral halves of newt iris. Foci of depigmented cells appeared within the cell reaggregates, regardless of their origins, when the cell reaggregates were cultured with FGF2 or FGF4. In contrast, only the depigmented cells in the dorsal iris cell reaggregates underwent extensive proliferation and developed a lens with the synthesis of lens-specific crystallins, recapitulating the normal lens regeneration. On the other hand, neither FGF8, FGF10, EGF, VEGF, nor IGF promoted lens development from iris cell reaggregates. Consistent with the FGF-specific action, FGFR-specific inhibitor SU5402 suppressed the lens development from the cultured cell reaggregates. These results demonstrated that FGF2 or FGF4 is essential for the in vitro lens regeneration from the pigmented cells of the dorsal iris. In addition, these findings indicated that unequal competence in the dorsal and ventral iris to FGF2/4 contributes to the difference in lens forming ability between them. PMID- 12076338 TI - Ectoderm gene activation in sea urchin embryos mediated by the CCAAT-binding factor. AB - Transcriptional enhancers are short stretches of DNA that function to achieve highly specific patterns of gene expression. To identify the mechanisms by which enhancers achieve their specificity, we made use of an enhancer from the aboral ectoderm-specific spec2a gene of the sea urchin Strongylocentrotus purpuratus. The spec2a enhancer contains five cis-regulatory elements within 78 base pairs that interact with five distinct DNA-binding proteins to confer aboral ectoderm expression. Here, we present an analysis of the sea urchin CCAAT binding factor (CBF), which binds to a CCAAT motif within the spec2a enhancer. S. purpuratus CBF and SpOtx, a ubiquitously expressed factor, act together at closely placed cis regulatory elements to mediate spec2a transcription in the ectoderm. SpCBF was the sole factor that bound to the spec2a CCAAT element, and two of the three subunits that make up the CBF holoprotein were cloned and shown to have high sequence conservation with their vertebrate orthologs. Based on its involvement in the regulation of several other sea urchin genes, SpCBF appears to be a major transcription factor in the sea urchin embryo for positive regulation of ectoderm gene expression. In addition to its role in vertebrate cell growth and proliferation, our results indicate that CBF also functions at the early stages of germ layer formation, namely ectoderm differentiation. PMID- 12076339 TI - Non-thermal activation of the hsp27/p38MAPK stress pathway by mobile phone radiation in human endothelial cells: molecular mechanism for cancer- and blood brain barrier-related effects. AB - We have examined whether non-thermal exposures of cultures of the human endothelial cell line EA.hy926 to 900 MHz GSM mobile phone microwave radiation could activate stress response. Results obtained demonstrate that 1-hour non thermal exposure of EA.hy926 cells changes the phosphorylation status of numerous, yet largely unidentified, proteins. One of the affected proteins was identified as heat shock protein-27 (hsp27). Mobile phone exposure caused a transient increase in phosphorylation of hsp27, an effect which was prevented by SB203580, a specific inhibitor of p38 mitogen-activated protein kinase (p38MAPK). Also, mobile phone exposure caused transient changes in the protein expression levels of hsp27 and p38MAPK. All these changes were non-thermal effects because, as determined using temperature probes, irradiation did not alter the temperature of cell cultures, which remained throughout the irradiation period at 37 +/- 0.3 degrees C. Changes in the overall pattern of protein phosphorylation suggest that mobile phone radiation activates a variety of cellular signal transduction pathways, among them the hsp27/p38MAPK stress response pathway. Based on the known functions of hsp27, we put forward the hypothesis that mobile phone radiation-induced activation of hsp27 may (i) facilitate the development of brain cancer by inhibiting the cytochrome c/caspase-3 apoptotic pathway and (ii) cause an increase in blood-brain barrier permeability through stabilization of endothelial cell stress fibers. We postulate that these events, when occurring repeatedly over a long period of time, might become a health hazard because of the possible accumulation of brain tissue damage. Furthermore, our hypothesis suggests that other brain damaging factors may co-participate in mobile phone radiation-induced effects. PMID- 12076340 TI - Pulse pressure in tests improves the prediction of left ventricular mass: 10 years of follow-up. AB - Literature does not agree with the usefulness of exercise blood pressure (BP) in predicting hypertension or target organ damage. In this prospective 10 years of follow-up, we evaluated if exaggerated BP responses to tests may improve the prediction of left ventricular mass index (LVMI). At baseline, BP was recorded by casual measurements, and during tests using intra-arterial monitoring. The subjects were 97 healthy, untreated 35- to 45-year-old-men (34 normotensive, 29 borderline hypertensive, and 34 mild hypertensive). At 10-year follow-up, echocardiography was performed to 86 (89%) of them. Subjects not taking antihypertensive medication (n = 66) were included in the prediction of LVMI(g m 2). Echocardiography data at baseline was available from 70 (72%) of the subjects, of whom 52 did not use antihypertensive medication at follow-up. Pulse pressure (PP) at supine test (r = 0.337, P = 0.006), PP at dynamic exercise last work load (r = 0.332, P = 0.006), and PP after dynamic exercise (r = 0.316, P = 0.010) were the best BP variables achieved in tests in predicting future LVMI of the 66 subjects. Casual BP did not significantly correlate with future LVMI. The best model in predicting LVMI included PP achieved after dynamic exercise, family history of hypertension, and body mass index (BMI) (adj.R2 = 0.207). Baseline LVMI correlated significantly with future LVMI only among the 52 unmedicated subjects (r = 0.508, P<0.0001). The predictive value of baseline LVMI on future LVMI among them (adj.R2 = 0.243) was best improved by PP achieved in supine test and age (adj.R2 = 0.350). In conclusion, BP measurements during tests improved the prediction of LVMI compared with casual BP. For the first time, the pulsatile component of BP in tests was found to be the most significant BP parameter in predicting future LVMI. PMID- 12076341 TI - Age-related reductions in appendicular skeletal muscle mass: association with habitual aerobic exercise status. AB - Appendicular skeletal muscle mass decreases with advancing age, and this is thought to be a key factor for reductions in functional independence in the elderly. We determined the potential modulatory influence of regular aerobic exercise on the age-related reductions in appendicular muscle mass. In the present cross-sectional study, we studied a total of 131 healthy men aged 20-79 years. For at least the previous 2 years subjects were either sedentary or endurance-trained. Appendicular skeletal muscle mass was measured using dual energy X-ray absorptiometry. There were no significant group differences in height. Body mass and body surface area were lower in endurance-trained compared with sedentary men (P<0.05). In general, body fat increased with age in both groups, but was lower in endurance-trained compared with sedentary men at any age group (P<0.05). Lean body mass and maximal oxygen consumption decreased with age (P<0.05). Irrespective of expression (absolute, relative to body surface area, or relative to body mass), arm, leg, and total limb muscle mass generally decreased with age. Absolute appendicular muscle mass was not different between the two activity groups. When expressed relative to body mass or body surface area, appendicular muscle mass in the endurance-trained middle-aged and older men was significantly higher than their sedentary peers. The rate of decline in appendicular muscle mass with age was similar between the two activity groups. The results of the present cross-sectional study indicate that endurance-trained men had significantly higher appendicular skeletal muscle mass relative to body mass or body size compared with their sedentary peers. This may contribute, at least in part, to the lower incidence of functional disability observed in middle aged and older men who exercise regularly. PMID- 12076343 TI - Validation of a newly developed B-mode image-processing technique versus wall tracking ultrasound for the study of wall mechanics in small-calibre arteries. AB - PURPOSE: To validate a newly developed image-processing technique for the assessment of arterial wall compliance and distensibility from non-invasive B mode ultrasound compared with the invasive wall-tracking technique. MATERIALS AND METHODS: Arterial wall compliance and distensibility coefficient were measured invasively by wall-tracking with an ultrasonic transducer implanted on the vessel wall, and non-invasively by automatic processing of B-mode ultrasound images, with a dedicated workstation and software (IO 3.1, IODP, Paris). Measurements were performed in the normal aorta of five animals, and upstream, at the stent level, and downstream from the stent in eight other animals (immediately after stenting in six, and 3 months later in four), for a total of 35 paired measurements. RESULTS: There was no significant difference between the two techniques for compliance but there was a significant difference in diameter (P<0.005) and distensibility (P<0.05) as external ultrasound measured the inner diameter, while wall-tracking measured the outer diameter. Agreement between the two methods as assessed by the Bland-Altman approach was acceptable for aortic diameter, compliance and distensibility. CONCLUSION: Automatic processing of B mode ultrasound images is a reliable non-invasive technique to assess the compliance of small-calibre arteries. PMID- 12076342 TI - Effects of endurance training on heart rate and blood pressure variability. AB - PURPOSE: To study the influences of a 1-year controlled, randomized endurance exercise training period on heart rate (HR) and blood pressure variability in a representative sample of Finnish men in their late middle age. METHODS AND RESULTS: Subjects were 140 sedentary men aged 53-63 years. The men were randomized into two identical groups: an intervention (EX) and a reference (CO) group. One hundred and twelve of them remained in the final analysis (EX: n=59, CO: n=53). EX trained for 30-60 min three to five times a week with the intensity of 40-60% of maximal oxygen consumption. In EX, 1 year of regular exercise training increased oxygen consumption at respiratory compensation threshold by 11% (P < or = 0.001) in a maximal cardiorespiratory test. Total power and very low frequency power of R-R interval variability (ms2) tended to increase in the EX group by 26 and 42% and to decrease in the CO group by 13 and 10% (interaction P<0.05 and P<0.01), respectively. There were no significant changes in blood pressure variability. CONCLUSION: Regular low- to moderate-intensity exercise training could retard the declining tendency in cardiac autonomic nervous function in older men during 1 year. PMID- 12076344 TI - A single-blind placebo-controlled investigation into the analgesic effects of interferential currents on experimentally induced ischaemic pain in healthy subjects. AB - The aim of this single-blind placebo-controlled study was to examine the analgesic effects of interferential currents (IFC) on experimentally induced ischaemic pain. Ischaemic pain was induced using the submaximal effort tourniquet technique (SETT) and pain intensity was recorded using a visual analogue scale at 1-min intervals was used as the primary outcome measure. Following baseline recordings 30 healthy volunteers received either active IFC, sham IFC, or no treatment (10 subjects per group). Data were analysed by calculating the mean change in pain intensity at each 1-min interval by subtracting data during treatment from the baseline data. IFC was administered throughout the duration of the ischaemic pain test via four electrodes (quadripolar application) on the forearm. Active IFC delivered electrical currents at a 'strong but comfortable' intensity. A 'dummy' stimulator that delivered no current was used as sham IFC. Subjects in the no treatment control group were informed that the IFC device was not switched on. There were significant effects for Groups (P=0.04) which were attributed to a significant reduction in pain intensity for the IFC group when compared with sham and no-treatment control (P< or =0.05). There were no significant effects for Time (P=0.69) or Group-Time interaction (P=0.45). In conclusion, IFC produced significantly greater analgesia than sham and no treatment control groups under the present experimental conditions. PMID- 12076345 TI - Assessment of fluid balance by measurement of skin tissue thickness during clinical anaesthesia. AB - STUDY OBJECTIVE: To determine if measuring skin tissue thickness by a recently developed 10 MHz ultrasound scan may be used as a valuable parameter to guide fluid therapy and detect fluid shifts to the extravascular space during surgical procedures in addition to central venous pressure (CVP). STUDY DESIGN: Prospective, clinical and observational study. SETTING: An operation theatre of the Ear Nose Throat (ENT) department of an university hospital. PATIENTS: Following approval by the local ethics committee 12 otherwise healthy male patients undergoing ENT surgery for oropharyngeal carcinoma were involved in this study. The patients stayed nil per os for 10 h before induction of anaesthesia. INTERVENTIONS/MEASUREMENTS: Crystalline fluids (Ringer's solution) were supplied at a constant rate of 10 ml kg-1 bw h-1. Patients were kept in the supine position during surgery, no further interventions were performed. Additional to routine monitoring, tissue thickness (TT) of proximal pre-tibial skin and CVP were measured every 30 min, haematocrit was determined hourly for 5 h starting at t0. MAIN RESULTS: Haematocrit constantly declined during the observation period, showing a significant difference in t0 after 120 min. The increase in TT was strongly correlated with intraoperative positive fluid balance (r=0.96), while the course of CVP did not reflect the amount of fluid application comparably. CONCLUSIONS: Non-invasive determinations of skin tissue thickness by the presented ultrasonic device appears to give additional information on fluid intake and distribution during clinical anaesthesia. PMID- 12076346 TI - The internal jugular vein valve may have a significant role in the prevention of venous reflux: evidence from live and cadaveric human subjects. AB - INTRODUCTION: The internal jugular vein valve (IJVV), which is situated just above the termination of the internal jugular vein, is the only valve between the heart and the brain. This means that it plays a role in the prevention of cephalad flow of venous blood. If the IJVV is damaged or becomes incompetent, increase in intrapleural pressure could result in raised intracranial pressure. Additionally, the jugular venous pulse (JVP) is used clinically to estimate right atrial pressure, a functional IJVV may prevent accurate estimation of the JVP. OBJECTIVES: To describe the presence and the competence of the IJVV in post mortem and live human subjects. DESIGN - setting and methods: The anatomical appearance of the IJVV from 30 cadavers was studied. Competence was checked by measuring maximum hydrostatic pressure before reflux occurred through the valve. The function of the valve was evaluated in 25 live subjects using colour duplex scanning. RESULTS: The IJVV was present in all cadavers just before its termination (60 IJVVs from 30 subjects). The valve was bicuspid in most cases (93%). The competence of 41 IJVVs was checked of which only three (7%) were found to be incompetent. All IJVVs in live subjects were found to be competent. CONCLUSION: This study confirms that a functional IJVV is present just above the termination of the internal jugular vein. The IJVV may therefore prevent reflux of venous blood from the right atrium into the internal jugular vein. PMID- 12076347 TI - Do preceding vasoconstrictions influence the 'inspiratory gasp test'? AB - Diabetic or other neuropathies can affect the autonomic innervations of vessels and heart. Using non-invasive and inexpensive autonomic tests, one can detect autonomic neuropathy. One of these tests is the inspiratory gasp response (IGR). It appears as a short reduction of skin blood volume (SBV) following deep inspiration (DI). Unfortunately, because of a high degree of variability of the study results, the clinical merits of this test have been questioned. However, there remains vagueness about underlying mechanisms or pitfalls. The aim of the study was to identify preceding vasoconstrictor episodes as putative pitfalls in assessing IGR. With regard to the hypothesis that a preceding vasoconstrictor episode may affect the amplitude of the following one, we investigated SBV in fingertips of 25 healthy test subjects (14 females, 11 males; mean age 24 years), using the photoplethysmographic technique. After adaptation to environmental conditions the subjects were instructed to perform five DI in various time lags (90, 30, 20 and 180 s). We found that the shorter the time interval to a preceding IGR, the lower the amplitude of the following IGR. After an interval of 180 s, the amplitude increased again. We conclude that this newly observed phenomenon might explain the contradictory results of the IGR test found in some earlier studies. We suppose that reduced IGR may depend on alterations in peripheral norepinephrinergic transmission processes (e.g. emptying of norepinephrine reservoirs). In clinical studies it should be ruled out that a diagnostically assessed IGR is influenced by a preceding vasoconstrictor episode. PMID- 12076348 TI - In vivo quantitative near-infrared spectroscopy in skeletal muscle during incremental isometric handgrip exercise. AB - The aim of this study was to investigate the performance of in vivo quantitative near-infrared spectroscopy (NIRS) in skeletal muscle at various workloads. NIRS was used for the quantitative measurement of O2 consumption (mVO2) in the human flexor digitorum superficialis muscle at rest and during rhythmic isometric handgrip exercise in a broad range of work intensities (10-90% MVC=maximum voluntary contraction force). Six subjects were tested on three separate days. No significant differences were found in mVO2 measured over different days with the exception of the highest workload. The within-subject variability for each workload measured over the three measurements days ranged from 15.7 to 25.6% and did not increase at the high workloads. The mVO2 was 0.14 +/- 0.01 mlO2 min-1 100 g-1 at rest and increased roughly 19 times to 2.68 +/- 0.58 mlO2 min-1 100 g-1 at 72% MVC. These results show that local muscle oxygen consumption at rest as well as during exercise at a broad range of work intensities can be measured reliably by NIRS, applied to a uniform selected subject population. This is of great importance as direct local measurement of mVO2 during exercise is not possible with the conventional techniques. The method is robust enough to measure over separate days and at various workloads and can therefore contribute to a better understanding of human physiology in both the normal and pathological state of the muscle. PMID- 12076349 TI - Insulin inhalation with absorption enhancer at meal-times results in almost normal postprandial insulin profiles. AB - BACKGROUND: Conventional insulin therapy with subcutaneous injections of regular insulin at meal-times result in plasma insulin peaks that are lower and appear later than meal related insulin peaks in healthy individuals. The present study was designed in order to evaluate the resulting insulin concentrations in peripheral blood after inhalation of micro crystalline human insulin together with an absorption enhancer [dioctyl sodium sulphosuccinate (DOSS)] via a powder inhaler. METHODS: Ten insulin dependent middle-aged non-obese diabetic patients (mean diabetes duration 21 years) were included. Blood samples for glucose and insulin were taken immediately before and 13 times, up to 300 min, after insulin inhalation. The mass median aerodynamic diameter of the particles was 3.2 microm. The inhaled insulin dose was 39 U. RESULTS: Within 5 min after the end of the 2 min inhalation procedure the mean increase of insulin was 7.0 microU ml-1, and the mean maximum concentration, 12.1 microU ml-1, was reached between 20 and 30 min. There was then a slow decline until base-line was reached after around 210 min and there were no adverse events. CONCLUSIONS: Inhalation of a mixture of 39 U of insulin and enhancer resulted in a rapid plasma insulin peak with a slow decline, similar to the normal postprandial insulin profile. PMID- 12076350 TI - Lumbar bone mineral content and density measured using a Lunar DPX densitometer in healthy full-term infants during the first year of life. AB - This cross-sectional study provides values for lumbar spine bone mineral content (BMC) and density (BMD) in 41 healthy full-term born Finnish infants, 19 boys and 22 girls, during the first year of life measured by dual energy X-ray absorptiometry (DXA) using the Lunar DPX densitometer. Lumbar BMC correlated with the weight (r=0.733; P=0.000), length (r=0.677; P=0.000), standardized length (r=0.315; r=0.045) and age at examination (r=0.314; P=0.045), and with the bone area (r=0.736; P=0.000). Infants with < or =-1 SD scores for lengths at examination had significantly lower BMC values [mean (SD); 1.79 (0.66) g] than infants with SD scores above -1 SD [2.27 (0.46) g] (P=0.011). Exclusive breast feeding did not correlate with the lumbar BMC values (r=-0.039; P=0.811). No differences were found in lumbar spine BMC (P=0.097), BMD (P=0.254) and bone area (P=0.094) values between boys and girls. In order to determine the predictive value of the anthropometric measurements on lumbar BMC, stepwise multiple regression analysis were performed, bone area and present weight were the only independent variables which explained 67.6% of the variance in the BMC values. The present cross-sectional data imply that, in healthy term infants, patterns of relative linear growth during the first year of life are related to the lumbar BMC values. In future, careful longitudinal measurements of linear growth are needed to study connections between growth patterns and bone mineral status in infancy. PMID- 12076351 TI - Skeletal muscle mitochondrial ATP production rate and walking performance in peripheral arterial disease. AB - This study tested the hypotheses that skeletal muscle mitochondrial ATP production rate (MAPR) is impaired in patients with peripheral arterial disease (PAD) and that it relates positively to their walking performances. Seven untrained patients, eight exercise-trained patients and 11 healthy controls completed a maximal walking test and had muscle sampled from the gastrocnemius medialis muscle. Muscle was analysed for its MAPR in the presence of pyruvate, palmitoyl-L-carnitine or both, as well as citrate synthase (CS) activity. MAPRs were not different between untrained PAD and controls. In contrast, MAPRs (pyruvate) were significantly higher in trained PAD vs. controls. MAPR (pyruvate combinations) was also significantly higher in trained than untrained PAD muscle. MAPR and CS activity were highly correlated with walking performance in patients, but not in controls. These data do not support the hypothesis that isolated mitochondria are functionally impaired in PAD and demonstrate that the muscle mitochondrial capacity to oxidize carbohydrate is positively related to walking performance in these patients. PMID- 12076352 TI - The quality of spirometric examinations in Finland: results from a national questionnaire survey. AB - National treatment programmes for asthma and chronic obstructive pulmonary disease (COPD) have in the past few years increased the need for spirometry in Finland. The purpose of this study was to determine, by means of a national questionnaire survey, the quality and number of spirometric examinations performed in Finland in 1998, the year when the national COPD programme was initiated. We estimated that 395 000-425 000 spirometric examinations were performed in Finland in 1998. The mean quality of spirometry was evaluated as moderate, and the quality index developed for this study indicated an average of scores of 67 (max 100), ranging between 43 and 88 in the whole country. The most common qualitative insufficiencies in spirometric examinations were found in preparation of the subject for the examination, performance of the examination, recognition of a successful spirometric curve and evaluation of reproducibility of the examination. Furthermore, the time reserved in the reports for performing the examination was often very short. However, in comparison with the previous questionnaire survey in Finland (in 1990), the quality of spirometry had markedly improved. PMID- 12076353 TI - Can we speed up the online publishing process? And who will pay for it, anyway? PMID- 12076354 TI - Antitumor effect of lysine-isopeptides. AB - Isopeptides (ϵ-peptides) of lysine, with a given Mw and low polydispersity (10-400 units), were synthesized to study the relationship between their chemical structure and biological effect. The designed compounds were of high purity, low polydispersity and high stereochemical purity. The effect of the compounds was tested on a human erythroleukemia cell line (K-562) and on four transplantable mouse tumors (L1210 lymphoid leukemia, P38 macrophage derived tumor, Ehrlich ascites carcinoma, Lewis lung tumor /LLT/). In case of the L1210 and P388 tumors and the Ehrlich carcinoma, survival of the animals was used as an indicator of the effect. In case of the Lewis lung tumor, the number and size of metastases in the lung and/or liver of treated and untreated mice were used as indicators. The polymers of polymerisation degree 80-120 (Mw 10.2-15.4 KD) showed the strongest antiproliferative effect both on K562 cells and the tumors growing in vivo. This effect was manifest with a significantly higher survival rate as compared to the control (L1210, P38, Ehrlich ascites), furthermore, by a decrease in the number and size of liver and lung metastases (LLT). PMID- 12076356 TI - Purification and partial characterization of bacillocin 490, a novel bacteriocin produced by a thermophilic strain of Bacillus licheniformis. AB - BACKGROUND: Applications of bacteriocins as food preservatives have been so far limited, principally because of their low antimicrobial activity in foods. Nisin is the only bacteriocin of significant use, but applications are restricted principally because of its very low activity at neutral or alkaline pH. Thus the isolation of new bacteriocins active in foods is desirable. RESULTS: We isolated a Bacillus licheniformis thermophilic strain producing a bacteriocin with some novel features, named here bacillocin 490. This bacteriocin was inactivated by pronase E and proteinase K and was active against closely related Bacillus spp. both in aerobic and in anaerobic conditions. Bactericidal activity was kept during storage at 4 degrees C and was remarkably stable in a wide pH range. The bacteriocin was partially purified by elution after adhesion to cells of the food isolated strain Bacillus smithii and had a rather low mass (2 KDa). Antimicrobial activity against B. smithii was observed also when this organism was grown in water buffalo milk. CONCLUSIONS: Bacillocin 490 is a novel candidate as a food anti-microbial agent since it displays its activity in milk, is stable to heat treatment and during storage, is active in a wide pH range and has bactericidal activity also at high temperature. These features may allow the use of bacillocin 490 during processes performed at high temperature and as a complementary antimicrobial agent of nisin against some Bacillus spp. in non-acidic foods. The small size suggests its use on solid foods. PMID- 12076355 TI - Stress responses and replication of plasmids in bacterial cells. AB - Plasmids, DNA (or rarely RNA) molecules which replicate in cells autonomously (independently of chromosomes) as non-essential genetic elements, play important roles for microbes grown under specific environmental conditions as well as in scientific laboratories and in biotechnology. For example, bacterial plasmids are excellent models in studies on regulation of DNA replication, and their derivatives are the most commonly used vectors in genetic engineering. Detailed mechanisms of replication initiation, which is the crucial process for efficient maintenance of plasmids in cells, have been elucidated for several plasmids. However, to understand plasmid biology, it is necessary to understand regulation of plasmid DNA replication in response to different environmental conditions in which host cells exist. Knowledge of such regulatory processes is also very important for those who use plasmids as expression vectors to produce large amounts of recombinant proteins. Variable conditions in large-scale fermentations must influence replication of plasmid DNA in cells, thus affecting the efficiency of recombinant gene expression significantly. Contrary to extensively investigated biochemistry of plasmid replication, molecular mechanisms of regulation of plasmid DNA replication in response to various environmental stress conditions are relatively poorly understood. There are, however, recently published studies that add significant data to our knowledge on relations between cellular stress responses and control of plasmid DNA replication. In this review we focus on plasmids derived from bacteriophage lambda that are among the best investigated replicons. Nevertheless, recent results of studies on other plasmids are also discussed shortly. PMID- 12076357 TI - Long-term experience with implanted intrathecal drug administration systems for failed back syndrome and chronic mechanical low back pain. AB - BACKGROUND: Continuous intrathecal drug delivery has been shown in open studies to improve pain and quality of life in those with intractable back pain who have had spinal surgery. There is limited data on long term effects and and even less for patients with mechanical back pain without prior spinal surgery. METHODS: We have investigated spinal drug administration systems for patients with failed back syndrome and chronic mechanical low back pain by patient questionnaire study of the efficacy of this therapy and a case notes review. RESULTS: 36 patients (97% of 37 approached) completed questionnaires, 24 with failed back syndrome and 12 with chronic mechanical low back pain. Recalled pre-treatment levels with current post-treatment levels of pain and a range of quality of life measures (recorded on 11-point numerical rating scales) were compared. Pain improved significantly in both groups (Wilcoxan signed ranks test, p < 0.005). The majority of quality of life measures improved significantly in the failed back syndrome group (Wilcoxan signed ranks test, p < 0.005) although work interruption and the effect of pain on sex life did not change. There was a trend towards improvement in the majority of quality of life measures in the mechanical back pain group but this did not reach statistical significance due to the smaller numbers in this cohort (p > 0.005, Wilcoxan signed ranks test with Bonferroni correction).Diamorphine was used in all 37 patients, bupivacaine in 32, clonidine in 27 and baclofen in 3. The mean dose of diamorphine increased for the first 2 years but did not change 2-6 years post implant, averaging 4.5 mg/day. Revision surgery was required in 24% of cases, but reduced to 12% in the later years of our experience. CONCLUSIONS: We conclude that spinal drug administration systems appear to be of benefit in alleviating pain in the failed back syndrome and chronic mechanical low back pain but need to be examined prospectively. PMID- 12076375 TI - Acupuncture for smoking cessation. AB - BACKGROUND: Acupuncture and related techniques are promoted as a treatment for smoking cessation in the belief that they may reduce nicotine withdrawal symptoms. OBJECTIVES: The objective of this review is to determine the effectiveness of acupuncture and the allied therapies of acupressure, laser therapy and electrostimulation, in smoking cessation in comparison with: a) sham treatment, b) other interventions, or c) no intervention. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register, Cochrane Controlled Trials Register, Medline, Embase, BIOSIS Previews, PsycINFO, Science and Social Sciences Citation Index, AMED and CISCOM. Date of last search January 2002. SELECTION CRITERIA: Randomised trials comparing a form of acupuncture, acupressure, laser therapy or electrostimulation with either sham treatment, another intervention or no intervention for smoking cessation. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of smokers recruited, the nature of the acupuncture and control procedures, the outcome measures, method of randomisation, and completeness of follow-up. We assessed abstinence from smoking at the earliest time-point (before 6 weeks), at six months and at one year or more follow-up in patients smoking at baseline. We used the most rigorous definition of abstinence for each trial, and biochemically validated rates if available. Those lost to follow-up were counted as continuing to smoke. Where appropriate, we performed meta-analysis using a fixed effects model. MAIN RESULTS: We identified 22 studies. Acupuncture was not superior to sham acupuncture in smoking cessation at any time point. The odds ratio (OR) for early outcomes was 1.22 (95% confidence interval 0.99 to 1.49); the OR after 6 months was 1.50 (95% confidence interval 0.99 to 2.27) and after 12 months 1.08 (95% confidence interval 0.77 to 1.52). Similarly, when acupuncture was compared with other anti-smoking interventions, there were no differences in outcome at any time point. Acupuncture appeared to be superior to no intervention in the early results, but this difference was not sustained. The results with different acupuncture techniques do not show any one particular method (i.e. auricular acupuncture or non-auricular acupuncture) to be superior to control intervention. Based on the results of single studies, acupressure was found to be superior to advice; laser therapy and electrostimulation were not superior to sham forms of these therapies. REVIEWER'S CONCLUSIONS: There is no clear evidence that acupuncture, acupressure, laser therapy or electrostimulation are effective for smoking cessation. PMID- 12076376 TI - Interventions for helping patients to follow prescriptions for medications. AB - BACKGROUND: People who are prescribed self-administered medications typically take less than half the prescribed doses. Efforts to assist patients with adherence to medications might improve the benefits and efficiency of health care, but also might increase its adverse effects. OBJECTIVES: To update a review summarising the results of randomised controlled trials (RCTs) of interventions to help patients follow prescriptions for medications for medical problems, focusing on trials that measured both adherence and clinical outcomes. SEARCH STRATEGY: Computerised searches to August 2001 in MEDLINE, CINAHL, The Cochrane Library, International Pharmaceutical Abstracts (IPA) PsychInfo, and Sociofile; bibliographies in articles on patient adherence; articles in the reviewers' personal collections; and contact with authors of original and review articles on the topic. SELECTION CRITERIA: Articles were selected if they reported an unconfounded RCT of an intervention to improve adherence with prescribed medications, measuring both medication adherence and treatment outcome, with at least 80% follow-up of each group studied and, for long-term treatments, at least six months follow-up for studies with positive initial findings. DATA COLLECTION AND ANALYSIS: Information on study design features, interventions and controls, and results were extracted by one reviewer and confirmed by at least one other reviewer. The studies were too disparate to warrant meta-analysis. MAIN RESULTS: For short-term treatments, one of three interventions reported in three RCTs showed an effect on both adherence and clinical outcome. Eighteen of 36 interventions for long-term treatments reported in 30 RCTs were associated with improvements in adherence, but only 16 interventions led to improvements in treatment outcomes. Almost all of the interventions that were effective for long term care were complex, including combinations of more convenient care, information, reminders, self-monitoring, reinforcement, counselling, family therapy, and other forms of additional supervision or attention by a health care provider (physician, nurse, pharmacist or other). Even the most effective interventions did not lead to large improvements in adherence and treatment outcomes. Two studies showed that telling patients about adverse effects of treatment did not affect their adherence. REVIEWER'S CONCLUSIONS: The full benefits of medications cannot be realised at currently achievable levels of adherence. Current methods of improving adherence for chronic health problems are mostly complex and not very effective. Innovations to assist patients to follow medication prescriptions are needed. PMID- 12076377 TI - Routine versus selective antifungal administration for control of fungal infections in patients with cancer. AB - BACKGROUND: Systemic fungal infection is considered to be an important cause of morbidity and mortality in cancer patients, particularly those with neutropenia. Antifungal drugs are often given prophylactically, or to patients with persistent fever. OBJECTIVES: The objective of this review was to assess the effect of antifungal drugs in cancer patients with neutropenia. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register and MEDLINE (November 2001) and the reference lists of articles. We searched the proceedings of the ICAAC (from 1990 to 2001, General Meeting of the ASM (from 1990 to 2001), and the European Congress of Clinical Microbiology and Infectious Diseases (1995 to 2001) and contacted researchers in the field. SELECTION CRITERIA: Randomised trials of amphotericin B, fluconazole, ketoconazole, miconazole, or itraconazole compared with placebo or no treatment in cancer patients with neutropenia. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial eligibility, methodological quality and abstracted data. MAIN RESULTS: Thirty trials involving 4094 patients were included. Prophylactic or empirical treatment with antifungals as a group had no statistically significant effect on mortality (relative risk 0.95, 95% confidence interval 0.82 to 1.11). The relative risk was smallest for amphotericin B, 0.73 (0.52 to 1.03) (P=0.08). In another review, three trials compared intravenous lipid soluble amphotericin B (AmBisome) with smaller doses of standard intravenous amphotericin B; the relative risk was 0.74 (0.52 to 1.07). Taken together, these results indicate that intravenous amphotericin B might decrease mortality. In contrast, trials with fluconazole, ketoconazole, miconazole and itraconazole failed to find an effect on mortality. The incidence of invasive fungal infection decreased significantly with administration of amphotericin B (relative risk 0.39, 95% CI 0.20 to 0.76), fluconazole (0.39, 0.27 to 0.57) and itraconazole (0.51, 0.27 to 0.96), but not with miconazole or ketoconazole. REVIEWER'S CONCLUSIONS: Intravenous amphotericin B is the only antifungal agent for which there is evidence suggesting that it might reduce mortality. It should therefore be preferred when prophylactic or empirical antifungal therapy in cancer patients with neutropenia is considered indicated. PMID- 12076378 TI - Holding chambers versus nebulisers for beta-agonist treatment of acute asthma. AB - BACKGROUND: In acute asthma inhaled beta-agonists are often administered to relieve bronchospasm by wet nebulisation, but some have argued that metered-dose inhalers with a holding chamber (spacer) can be equally effective. In the community setting nebulisers are more expensive, require a power source and need regular maintenance. OBJECTIVES: There is controversy as to whether wet nebulisers are better than metered dose inhalers with holding chambers to deliver beta2-agonist medications for acute asthma. Comparisons of hospital and home use are also of interest. The objective of this review was to assess the effects of holding chambers compared to nebulisers for the delivery of beta2-agonists for acute asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Randomised trials in adults and children (from two years of age) with asthma, where holding chamber beta2-agonist delivery was compared with wet nebulisation. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied study inclusion criteria (one reviewer for the first version of the review), extracted the data and assessed trial quality. Missing data were obtained from the authors or estimated. MAIN RESULTS: This review has been updated in 2001 to include 4 new trials and has now analysed 880 children and 444 adults included in 21 trials. Method of delivery of beta2-agonist did not appear to affect hospital admission rates. In adults, the relative risk of admission for holding chamber versus nebuliser was 0.88 (95% confidence interval 0.56 to 1.38). The relative risk for children was 0.65 (95% confidence interval 0.4 to 1.06). One study in children found a significantly shorter length of stay in the emergency department when the holding chamber was used, with a weighted mean difference of -0.62 hours, 95% confidence interval -0.84 to -0.40 hours. Adults' length of stay in the emergency department was similar for the two delivery methods. Peak flow and forced expiratory volume were also similar for the two delivery methods. Pulse rate was lower for holding chamber in children, weighted mean difference -7.8% baseline (95% confidence interval -10.2 to -5.3). REVIEWER'S CONCLUSIONS: Metered-dose inhalers with holding chamber produced outcomes that were at least equivalent to nebuliser delivery. Holding chambers may have some advantages compared to nebulisers for children with acute asthma. PMID- 12076379 TI - Corticosteroids for acute ischaemic stroke. AB - BACKGROUND: Much of the brain swelling in ischaemic stroke is due to cytotoxic oedema, which is related to cell membrane dysfunction. Early treatment with corticosteroids may help reduce the swelling and improve the outcomes after a stroke. OBJECTIVES: The objective of this review was to assess the effect of corticosteroids in acute presumed ischaemic stroke. SEARCH STRATEGY: We searched the Cochrane Stroke Group trials register (last searched: 10 April 2001) and contacted investigators in the field. SELECTION CRITERIA: Published randomised trials comparing corticosteroids with placebo or control in people with acute (presumed or definite) ischaemic stroke. Trials were included if treatment began within 48 hours of stroke onset and if clinical outcome was assessed. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied the inclusion criteria, assessed trial quality and extracted the data. MAIN RESULTS: Seven trials involving 453 people were included. Details of trial quality that may relate to bias were not available from most trials. No difference was shown in the odds of death within one year (odds ratio 1.08, 95% confidence interval 0.68 to 1.72). Treatment did not appear to improve functional outcome in survivors. Six trials reported neurological impairment but pooling the data was impossible because no common scale or time interval was used. The results were inconsistent between individual trials. The only adverse effects reported were small numbers of gastrointestinal bleeds, infections and deterioration of hyperglycaemia across both groups. Results unchanged since last update. REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate corticosteroid treatment for people with acute presumed ischaemic stroke. Conclusions unchanged since last update. PMID- 12076380 TI - Electroconvulsive therapy for schizophrenia. AB - BACKGROUND: Electroconvulsive therapy (ECT) involves the induction of a seizure (fit) for therapeutic purposes by the administration of a variable frequency electrical stimulus shock via electrodes applied to the scalp. The effects of its use in people with schizophrenia are unclear. OBJECTIVES: To determine whether electroconvulsive therapy (ECT) results in clinically meaningful benefit with regard to global improvement, hospitalisation, changes in mental state, behaviour and functioning for people with schizophrenia, and whether variations in the practical administration of ECT influences outcome. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1982-1996), EMBASE (1980-1996), MEDLINE (1966 2001), PsycLIT (1974-1996),SCISEARCH (1996) and the Cochrane Schizophrenia Group's Register (July 2001) were undertaken. The references of all identified studies were also inspected and authors contacted. SELECTION CRITERIA: All randomised controlled clinical trials that compared ECT with placebo, 'sham ECT', non-pharmacological interventions and antipsychotics, and different schedules and methods of administration of ECT for people with schizophrenia, schizoaffective disorder or chronic mental disorder. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality rated and data extracted. For dichotomous data, relative risks (RR) were estimated, with the 95% confidence intervals (CI). Where possible, the number needed to treat statistic (NNT) was calculated. Analysis was by intention-to-treat. Normal continuous data was summated using the weighted mean difference (WMD). Scale data was presented for only those tools that had attained pre-specified levels of quality. Tests of heterogeneity and for publication bias were undertaken. MAIN RESULTS: This review includes 24 trials with 46 reports. When ECT is compared with placebo or sham ECT, fewer people remain unimproved in the real ECT group (n=400, RR fixed 'not globally improved in the short term' 0.77 CI 0.6 to 0.9, chi-square 13.46 df=8 p=0.1). Removal of the one study with clearly heterogeneous results causes a change in the findings (n=380, 8 RCTs, RR fixed 0.83 CI 0.7 to 1.01), as does removal of a clinically heterogeneous trial (n=370, 8 RCTs, RR fixed 0.74 CI 0.6 to 0.9, chi-square 10.97 df=7 p=0.14). There was a suggestion that ECT resulted in less relapses than sham ECT (n=47, 2 RCTs, RR fixed 0.26 CI 0.03 to 2.2), and a greater likelihood of being discharged from hospital (n=98, 1 RCT, RR fixed 0.59, CI 0.34 to 1.01). There is no evidence that this early advantage for ECT is maintained over the medium to long term. People treated with ECT did not drop out of treatment earlier than those treated with sham ECT (n=495, 14 RCTs, RR fixed 0.71 CI 0.33 to 1.52). Very limited data indicated that visual memory might decline after ECT compared with sham ECT (n=24, 1 RCT, WMD -14.0 CI -23 to -5); the results of verbal memory tests were equivocal. When ECT is directly compared with antipsychotic drug treatment (total n=419, 8 RCTs), results favour the medication group (n=175, 3 RCTs, RR fixed 'not improved at the end of ECT course' 2.18 CI 1.3 to 3.6). One small study suggested more memory impairment after a course of ECT combined with antipsychotics than with antipsychotics alone (n=20, MD serial numbers and picture recall -4.90 CI -0. 8 to -9), though this proved transient. When continuation ECT was added to antipsychotic drugs, the combination was superior to the use of antipsychotics alone (n=30, WMD Global Assessment of Functioning 19.1 CI 9.7 to 28.5), or CECT alone (n=30, WMD -20.3 CI -11.5 to 29.1). Unilateral and bilateral ECT were equally effective in terms of global improvement (n=78, 2 RCTs, RR fixed 'not improved at end of course of ECT' 0.79 CI 0.5 to 1.4). One trial showed a significant advantage for 20 treatments over 12 treatments for numbers globally improved at the end of the ECT course (n=43, RR fixed 2.53 CI 1.1 to 5.7). REVIEWER'S CONCLUSIONS: There is no evidence to clearly refute the use of ECT for people with schizophrenia. There is some limited evidence to support its use, particularly combined with antipsychotic drugs for those with schizophrenia who show limited response to medication alone. The research base for the use of ECT in people with schizophrenia is growing but, even after more than five decades of clinical use, is still inadequate. PMID- 12076381 TI - Antibiotics for gonorrhoea in pregnancy. AB - BACKGROUND: Neisseria gonorrhoeae can be transmitted from the mother's genital tract to the newborn during birth and can cause gonococcal ophthalmia neonatorum as well as systemic neonatal infection. It can also cause endometritis and pelvic sepsis in the mother. OBJECTIVES: The objective of this review was to assess the effects of antibiotic regimens in the treatment of genital infection with gonorrhoea during pregnancy with respect to neonatal and maternal morbidity. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register (November 2001) and the Cochrane Controlled Trials Register (The Cochrane Library, Issue 3, 2001) were searched. SELECTION CRITERIA: Randomised trials of one regimen of antibiotic versus another in pregnant women with culture confirmed genital gonococcal infection. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by one reviewer. MAIN RESULTS: Two trials involving 346 women were included. The only outcome included in these trials was the incidence of 'cure' assessed by bacterial culture. Failure to achieve 'microbiological cure' was similar for each antibiotic regimen: Amoxicillin plus probenecid compared with spectinomycin (odds ratio (OR) 2.40, 95% confidence interval (CI) 0.71-8.12), amoxicillin plus probenecid compared with ceftriaxone (OR 2.40, 95% CI 0.71-8.12) and ceftriaxone compared with cefixime (OR 1.22, 95% CI 0.16-9.04). Side effects were uncommon for all the tested regimens. REVIEWER'S CONCLUSIONS: The number of women included in each of the comparisons is small and therefore, although no differences were detected between the different antibiotic regimens, the trials were limited in their ability to detect important but modest differences. For women who are allergic to penicillin, this review provides some reassurance that treatment with ceftriaxone or spectinomycin appears to have similar effectiveness in producing microbiological cure. PMID- 12076382 TI - Prophylactic intravenous preloading for regional analgesia in labour. AB - BACKGROUND: Fetal heart rate changes are common following regional analgesia (epidural or spinal) during labour. Reduced uterine blood flow from maternal hypotension (low blood pressure) may contribute to this. Intravenous fluid preloading (volume expansion) may help to reduce maternal hypotension. Newer protocols using weaker solutions of local anaesthetic, and opioid only blocks, may reduce the need for preloading. OBJECTIVES: The objective of this review was to assess the effects of prophylactic intravenous fluid preloading prior to epidural analgesia during labour on maternal and fetal well-being. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register (November 2001) and the Cochrane Controlled Trials Register (The Cochrane Library, Issue 3, 2001) were searched. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing prophylactic intravenous preloading before epidural analgesia during labour with a control group (dummy or no preloading). DATA COLLECTION AND ANALYSIS: The reviewer assessed trial quality and extracted data. MAIN RESULTS: For high dose local anaesthetic blocks, one study involving 102 women was included. There was potential for considerable bias in this trial. Preloading with intravenous fluids was associated with a reduction in hypotension (relative risk 0.07, 95% confidence interval 0.01 to 0.53). It was also associated with a reduction in fetal heart rate abnormalities (relative risk 0.36, 95% confidence interval 0.16 to 0.83). No differences were detected in other perinatal and maternal outcomes. The trials in women receiving low-dose local anaesthetic (95 women) or opioid only blocks (30 women) demonstrated no statistically significant differences in maternal hypotension or fetal heart rate abnormality, but were too small to exclude the possibility of moderate effects with certainty. REVIEWER'S CONCLUSIONS: There are methodological limitations in the trials studied. However, preloading prior to high-dose local anaesthetic blocks may have beneficial fetal and maternal effects in healthy women. Further investigation of the effects in women receiving low-dose local anaesthetic or opioid only blocks, and the risks and benefits of intravenous preloading for women with pregnancy complications, is required. PMID- 12076383 TI - Aerobic exercise for women during pregnancy. AB - BACKGROUND: Physiological responses of the fetus (especially increase in heart rate) to single, brief bouts of maternal exercise have been documented frequently, suggesting that maternal exercise might have beneficial or adverse effects on pregnancy outcome. OBJECTIVES: The objective of this review was to assess the effects of advising healthy pregnant women to engage in regular (at least two to three times per week) aerobic exercise on physical fitness, labour and delivery, and the outcome of pregnancy. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched and researchers in the field were contacted. Date of last search: January 2002. SELECTION CRITERIA: Acceptably controlled comparisons of prescribed aerobic exercise programmes. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality and extracted data. Study authors were contacted for additional information. MAIN RESULTS: Ten trials involving 688 women were included. The trials were not of high methodologic quality. Five trials reported significant improvement in physical fitness in the exercise group, although differences in measures used to assess fitness prevent a quantitative pooling of results. Six trials reported on pregnancy outcomes. A statistically significant increased risk of preterm birth (relative risk (RR) = 2.29 [95% confidence interval 1.02 to 5.13]) in the exercise group is inconsistent with the absence of effect on mean gestational age [+0.0 (-0.4 to +0.4) weeks] and may have been biased by post-randomization exclusion of women with preterm labor. Results bearing on fetal growth (birth weight, length, and head circumference) were highly variable. One trial found improvements among exercising women in several aspects of self-reported body image, including muscle strength, energy level, and body build. REVIEWER'S CONCLUSIONS: Regular aerobic exercise during pregnancy appears to improve (or maintain) physical fitness and body image. Available data are insufficient to infer important risks or benefits for the mother or infant. PMID- 12076384 TI - Interventions to help external cephalic version for breech presentation at term. AB - BACKGROUND: Breech presentation places a fetus at increased risk. The outcome for the baby is improved by planned caesarean section compared with planned vaginal delivery. External cephalic version attempt reduces the chance of breech presentation at birth, but is not always successful. Tocolytic drugs to relax the uterus as well as other methods have been also used in an attempt to facilitate external cephalic version at term. OBJECTIVES: The objective of this review is to assess the effects of routine tocolysis, fetal acoustic stimulation, epidural or spinal analgesia and transabdominal amnioinfusion for external cephalic version at term on successful version and measures of pregnancy outcome. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group Trials Register (searched December 2001) and the Cochrane Controlled Trials Register (The Cochrane Library, Issue 4, 2001) were searched. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing routine versus selective or no tocolysis; fetal acoustic stimulation in midline fetal spine positions versus dummy or no stimulation; epidural or spinal analgesia versus no regional analgesia; or transabdominal amnioinfusion versus no amnioinfusion for external cephalic version at term. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed by the reviewer. MAIN RESULTS: In six trials, routine tocolysis was associated with fewer failures of external cephalic version (relative risk 0.74, 95% confidence interval 0.64 to 0.87). The reduction in non-cephalic presentations at birth was not statistically significant. Caesarean sections were reduced (relative risk 0.85, 95% confidence interval 0.72 to 0.99). Fetal acoustic stimulation in midline fetal spine positions was associated with fewer failures of external cephalic version at term (relative risk 0.17, 95% confidence interval 0.05 to 0.60). With epidural or spinal analgesia, external cephalic version failure, non-cephalic births and caesarean sections were reduced in two trials but not the other. The overall differences were not statistically significant. No randomised trials of transabdominal amnioinfusion for external cephalic version at term were located. REVIEWER'S CONCLUSIONS: Routine tocolysis appears to reduce the failure rate of external cephalic version at term. Although promising, there is not enough evidence to evaluate the use of fetal acoustic stimulation in midline fetal spine positions, nor of epidural or spinal analgesia. Large volume intravenous preloading may have contributed to the effectiveness demonstrated in two of the latter trials. No randomised trials of transabdominal amnioinfusion for external cephalic version at term were found. PMID- 12076385 TI - Short-term low-dose corticosteroids vs placebo and nonsteroidal antiinflammatory drugs in rheumatoid arthritis. AB - BACKGROUND: The effectiveness of low dose corticosteroids, equivalent to 15 mg prednisolone daily or less, in patients with rheumatoid arthritis has been questioned. We therefore performed a systematic review of trials which compared corticosteroids with placebo or non-steroidal, anti-inflammatory drugs. OBJECTIVES: To determine whether short-term (i.e. as recorded within the first month of therapy), oral low-dose corticosteroids (corresponding to a maximum of 15 mg prednisolone daily) is superior to placebo and non steroidal, antiinflammatory drugs in patients with rheumatoid arthritis. SEARCH STRATEGY: Medline Silverplatter, The Cochrane Controlled Trials Register, reference lists and a personal archive. Date of last search Nov 2001. SELECTION CRITERIA: This review is an update to a previous review which was peer-review and published in the Cochrane Library. All randomised studies comparing an oral corticosteroid (not exceeding an equivalent of 15 mg prednisolone daily) with placebo or a non steroidal, antiinflammatory drug were eligible if they reported clinical outcomes within one month after start of therapy. DATA COLLECTION AND ANALYSIS: Decisions on which trials to include were made independently by two observers based on the methods sections of the trials only. Standardised effect measures were used for the statistical analyses; the random effects model was used if P<0.10 for the test of heterogeneity. MAIN RESULTS: Ten studies, involving 320 patients, were included in the meta-analysis. Prednisolone had a marked effect over placebo on joint tenderness (standardised effect size 1.31, 95% confidence interval 0.78 to 1.83), pain (standardised effect size 1.75, 0.87 to 2.64) and grip strength (standardised effect size 0.41, 0.13 to 0.69). Measured in the original units, the differences were 12 tender joints (6 to 18) and 22 mm Hg (5 to 40) for grip strength. Prednisolone also had a greater effect than non steroidal, antiinflammatory drugs on joint tenderness (standardised effect size 0.63, 0.11 to 1.16) and pain (standardised effect size 1.25, 0.26 to 2.24), whereas the difference in grip strength was not significant (standardised effect size 0.31, 0.02 to 0.64). Measured in the original units, the differences were 9 tender joints (5 to 12) and 12 mm Hg (-6 to 31). The risk of adverse effects, also during moderate- and long-term use, seemed acceptable. REVIEWER'S CONCLUSIONS: Prednisolone in low doses (not exceeding 15 mg daily) may be used intermittently in patients with rheumatoid arthritis, particularly if the disease cannot be controlled by other means. Since prednisolone is highly effective, short-term placebo controlled trials studying the clinical effect of low-dose prednisolone or other oral corticosteroids are no longer necessary. PMID- 12076386 TI - Routine or selective carotid artery shunting for carotid endarterectomy (and different methods of monitoring in selective shunting). AB - BACKGROUND: Temporary interruption of cerebral blood flow during carotid endarterectomy can be avoided by using a shunt across the clamped section of the carotid artery. This may improve outcome. OBJECTIVES: The objective of this review was to assess the effect of routine versus selective, or never, shunting during carotid endarterectomy, and to assess the best method for selecting patients for shunting. SEARCH STRATEGY: For the original review the authors searched the Cochrane Stroke Group trials register, Medline (1966 to 1994), Embase (1980 to 1995) and Index to Scientific and Technical Proceedings (1980 to 1994). They also hand searched Annals of Surgery (1981 to 1995), British Journal of Surgery (1985 to 1995), European Journal of Vascular Surgery (1988 to 1995) and World Journal of Surgery (1978 to 1995). For the updated review, for the dates January 1994 - December 2000 we: 1. Repeated all these searches performed for the original review and developed more comprehensive search strategies for Medline and Embase. The Cochrane Stroke Group Trials Register was last searched in May 2001. 2. Hand searched the Journal of Vascular Surgery, Stroke, Annals of Vascular Surgery, American Journal of Surgery and Cardiovascular Surgery. 3. Hand searched the abstracts from the International Stroke Conference, AGM of the Vascular Surgical Society (UK), AGM of the Association of Surgeons of Great Britain and Ireland and the Annual Meeting of the Society for Vascular Surgery (USA). 4. Searched reference lists from all relevant trials All the authors of studies included in the initial review, and other authors known to have published relevant work, were contacted requesting information about further published or unpublished data. SELECTION CRITERIA: Randomised and quasi-randomised trials of routine shunting compared with no shunting or selective shunting, and trials that compared different shunting policies in patients undergoing carotid endarterectomy. DATA COLLECTION AND ANALYSIS: For the original review two reviewers independently performed the searches and applied the inclusion criteria. The data were extracted by one reviewer and double-checked. Trial quality was assessed. During the update, two reviewers independently performed the searches and applied the inclusion criteria. No new relevant randomised controlled trials were found. MAIN RESULTS: Despite recommendation from the original review that further studies were required, no new trials of adequate quality and fitting the inclusion criteria were found. The initial review included three trials. Two trials involving 590 patients compared routine shunting with no shunting. The other trial involving 131 patients compared shunting with a combination of electroencephalographic and carotid pressure measurement, with shunting by carotid pressure measurement alone. Allocation was adequately concealed in one trial, and one trial was quasi-randomised. Analysis was by intention-to-treat where possible. For routine versus no shunting, there was no significant difference in the rate of all stroke, ipsilateral stroke or death up to 30 days after surgery, although data were limited. There was no significant difference between the risk of ipsilateral stroke in patients selected for shunting with the combination of electroencephalographic and carotid pressure assessment compared to pressure assessment alone, although again the data were limited. REVIEWER'S CONCLUSIONS: When first published in 1995, this review concluded that the data available were too limited to either support or refute the use of routine or selective shunting in carotid endarterectomy. It was suggested that large scale randomised trials using no shunting as the control group were required. No one method of monitoring in selective shunting has been shown to produce better outcomes. No further prospective randomised or quasi randomised trials have been performed since then and the conclusions therefore remain unchanged. PMID- 12076387 TI - Ambulatory oxygen for chronic obstructive pulmonary disease. AB - BACKGROUND: Little is known about the effectiveness of ambulatory domicilary oxygen therapy. At present ambulatory oxygen in the UK is provided with small oxygen cylinders but in other countries such as the USA and Italy, liquid oxygen systems with higher oxygen carrying capacity are widely used. Both these systems are used without adequate evidence of their effectiveness. OBJECTIVES: To determine the effectiveness of long-term ambulatory domicilary oxygen therapy in patients with chronic obstructive pulmonary disease. SEARCH STRATEGY: The Cochrane Airways Group specialised trials register was searched. In addition, bibliographies of each trial retrieved were also searched for additional papers that may contain further studies. Authors of identified trials were contacted for other published and unpublished studies. SELECTION CRITERIA: Only randomised controlled trials in patients with chronic obstructive pulmonary disease were considered for inclusion. Trials must have randomised patients into long-term ambulatory oxygen therapy or placebo while at home. Ambulatory oxygen can be provided either through portable oxygen cylinders or with liquid oxygen canisters and the placebo group using compressed or liquid air. DATA COLLECTION AND ANALYSIS: Two reviewers assessed all potential trials independently. Data abstraction was completed by one reviewer and re-checked by the second reviewer. MAIN RESULTS: Only two studies met the inclusion criteria. These provided data on 70 patients. Statistically significant effects of oxygen were found in only one of the studies, a crossover trial involving 9 patients with severe hypoxia at rest. This study reported a reduction in minute ventilation at maximal exercise (WMD -11.00 L/min; 95%CI -17.53, -4.47L/min) and an increase in PaO2 at rest (WMD 17.00 mmHg; 95%CI 9.13,24.87 L/min) with oxygen therapy when compared to placebo. The other study recruited patients who did not have resting hypoxaemia. REVIEWER'S CONCLUSIONS: Evidence available to date does not allow any firm conclusions to be drawn concerning the effectiveness of ambulatory domicilary oxygen therapy in patients with COPD. Further studies are required in order to understand the role of ambulatory oxygen in the management of patients with COPD on long-term oxygen therapy. These studies should separate patients who desaturate from those who do not desaturate. PMID- 12076388 TI - Amphotericin B versus fluconazole for controlling fungal infections in neutropenic cancer patients. AB - BACKGROUND: Systemic fungal infection is considered to be an important cause of morbidity and mortality in cancer patients, particularly those with neutropenia. Antifungal drugs are often given prophylactically, or to patients with persistent fever. OBJECTIVES: To compare the effect of fluconazole and amphotericin B on morbidity and mortality in patients with cancer complicated by neutropenia. SEARCH STRATEGY: MEDLINE and Cochrane Library (November 2001). Letters, abstracts, and unpublished trials. The industry and authors were contacted. SELECTION CRITERIA: Randomised trials comparing fluconazole with amphotericin B. DATA COLLECTION AND ANALYSIS: Data on mortality, invasive fungal infection, colonisation, use of additional (escape) antifungal therapy and adverse effects leading to discontinuation of therapy were extracted by both authors independently. MAIN RESULTS: Sixteen trials (3760 patients, 341 deaths) were included. In 3 large 3-armed trials, results for amphotericin B were combined with results for nystatin in a "polyene" group. Because nystatin is an ineffective drug in these circumstances, this approach creates a bias in favour of fluconazole. Furthermore, most patients were randomised to oral amphotericin B, which is poorly absorbed and poorly documented. It was unclear whether there was overlap among the "polyene" trials. We were unable to obtain any information to clarify these issues from the trial authors or from Pfizer, the manufacturer of fluconazole. There were no significant differences in effect between fluconazole and amphotericin B, but the confidence intervals were wide. More patients dropped out of the study when they received amphotericin B, but as none of the trials were blinded, decisions on premature interruption of therapy could have been biased. Furthermore, amphotericin B was rarely given under optimal circumstances, with premedication to reduce infusion-related toxicity, slow infusion, and with potassium and magnesium supplements to prevent nephrotoxicity. REVIEWER'S CONCLUSIONS: Amphotericin B had been disfavoured in several of the trials through their design or analysis. Since intravenous amphotericin B is the only antifungal agent for which there is good evidence suggesting an effect on mortality and is considerably cheaper than fluconazole, it should be preferred. PMID- 12076389 TI - Antioxidant vitamin and mineral supplements for age-related macular degeneration. AB - BACKGROUND: It has been proposed that antioxidants may prevent cellular damage in the retina by reacting with free radicals produced in the process of light absorption. OBJECTIVES: The objective of this review is to assess the effects of antioxidant vitamin and/or mineral supplementation on the progression of age related macular degeneration. SEARCH STRATEGY: The Cochrane Controlled Trials Register - CENTRAL/CCTR, which contains the Cochrane Eyes and Vision Group specialised register (Cochrane Library Issue 3 2001), MEDLINE (1966 to August 2001), EMBASE (1980 to September 2001), the Science Citation Index, and the reference lists of relevant articles were searched. Investigators of included studies were contacted for further information. SELECTION CRITERIA: Randomised trials comparing an antioxidant vitamin and/or mineral supplement (alone or in combination) to control in people with age-related macular degeneration are included in this review. DATA COLLECTION AND ANALYSIS: The reviewer extracted data and assessed trial quality. Due to the variable methods of collecting and presenting outcome data, no statistical summary measure was calculated. MAIN RESULTS: Seven trials, which randomised 4119 people with signs of age-related macular degeneration, are included in this review. One unpublished trial of zinc supplementation (170 participants) is awaiting assessment. The majority of people (88%) were randomised in one trial that found a modest beneficial effect of antioxidant and zinc supplementation on progression to advanced age-related macular degeneration (odds ratio 0.72, 99% confidence interval 0.52 to 0.98). People supplemented with antioxidants and zinc were less likely to lose 15 or more letters of visual acuity (equivalent to a doubling of the visual angle) (odds ratio 0.79, 99% confidence interval 0.60 to 1.04). This effect was seen more strongly in people with moderate to severe disease. There were few events in people with early signs of the disease. The trial evaluated many safety outcomes, of which hospitalisation for genitourinary problems was more common in people taking zinc and yellowing of skin was more common in people taking antioxidant micronutrients. The other six trials in this review were small and the results were inconsistent. REVIEWER'S CONCLUSIONS: The evidence as to the effectiveness of antioxidant vitamin and mineral supplementation in halting the progression of age-related macular degeneration is dominated by one large trial that showed modest benefit in people with moderate to severe signs of the disease. There is no evidence at present that people with early signs of the disease should take supplementation, however, current studies are underpowered to answer that question. Long term harm from supplementation cannot be ruled out, particularly in smokers. The generalisability of these findings to other populations with different nutritional statuses is not known. Further large well-conducted randomised controlled trials in other populations are required. PMID- 12076390 TI - Chest physiotherapy for preventing morbidity in babies being extubated from mechanical ventilation. AB - BACKGROUND: Chest physiotherapy has been used to clear secretions and help lung ventilation in newborns who have needed mechanical ventilation for respiratory problems. However concerns about the safety of some forms of chest physiotherapy have been expressed. OBJECTIVES: To assess the effects of active chest physiotherapy on babies being extubated from mechanical ventilation for neonatal respiratory failure. SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Review Group was used. This included searches of electronic databases: Oxford Database of Perinatal Trials; Cochrane Controlled Trials Register (Cochrane Library Issue 4 2001); MEDLINE (1966-2001); and CINAHL (1982 2001), previous reviews including cross references, abstracts, conferences, symposia proceedings, expert informants and journal hand searching mainly in the English language. SELECTION CRITERIA: All trials utilising random or quasi-random patient allocation, in which active chest physiotherapy was compared with non active techniques (eg positioning and suction alone) or no intervention in the periextubation period. DATA COLLECTION AND ANALYSIS: Assessment of methodological quality and extraction of data for each included trial was undertaken independently by the authors. Data were extracted for the primary outcomes of postextubation lobar collapse, use of reintubation, duration of oxygen therapy, intracranial haemorrhage, cerebral cystic lesions, long term neurosensory impairment and death. Subgroup analysis was performed on different treatment frequencies and gestational age less than 32 weeks. Meta-analysis was conducted using a fixed effects model. Results are presented as relative risk (RR), risk difference (RD) and number needed to treat (NNT) for categorical data and mean difference (MD) for data measured on a continuous scale. All outcomes are reported with the use of 95% confidence intervals. MAIN RESULTS: In this review of four trials, two of which were carried out 15 & 23 years ago, no clear benefit of periextubation active chest physiotherapy can be seen. Active chest physiotherapy did not significantly reduce the rate of postextubation lobar collapse [RR 0.80 (95% CI 0.49,1.29)], though a reduction in the use of reintubation was shown in the overall analysis: RR 0.32 (95% CI 0.13,0.82); RD 7% (95% CI-13, -2); NNT 14 (95% CI 8, 50). There is insufficient information to adequately assess important short and longer term outcomes, including adverse effects. REVIEWER'S CONCLUSIONS: The results of this review do not allow development of clear guidelines for clinical practice. Caution is required when interpreting the possible positive effects of chest physiotherapy of a reduction in the use of reintubation and the trend for decreased post-extubation atelectasis as the numbers of babies studied are small, the results are not consistent across trials, data on safety are insufficient, and applicability to current practice may be limited. Further randomised controlled trials addressing the role of prophylactic active chest physiotherapy for neonates in the postextubation period may be unwarranted. PMID- 12076391 TI - Conservative interventions for treating distal radial fractures in adults. AB - BACKGROUND: Fracture of the distal radius is a common clinical problem particularly in elderly white women with osteoporosis. OBJECTIVES: To determine the most appropriate conservative treatment for fractures of the distal radius in adults. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register (January 2002), the Cochrane Controlled Trials Register (The Cochrane Library, Issue 4, 2001), MEDLINE (1966 to January 2002), EMBASE (1988 to 2002 Week 04), CINAHL (1982 to December Week 2 2001), the National Research Register (up to Issue 4, 2001) and reference lists of articles. No language restrictions were applied. SELECTION CRITERIA: Randomised or quasi-randomised clinical trials involving skeletally mature patients with a fracture of the distal radius, which compared commonly applied conservative interventions for fracture fixation. These included the application of an external support (plaster cast or brace) and fracture manipulation. DATA COLLECTION AND ANALYSIS: All trials, judged as fitting the selection criteria by both reviewers, were independently assessed by both reviewers for methodological quality. Data were extracted for anatomical, functional and clinical, including complications, outcomes. The trials were grouped into categories relating to cast position, extent and duration of immobilisation, use of braces, cast material and fracture manipulation. Although quantitative data from some trials are presented, the lack of good quality trials and trial heterogeneity inhibited pooling of results. MAIN RESULTS: Two trials were newly included in this update. In all, there are 33 trials, involving a total of 3664 mainly female and older patients, meeting the inclusion criteria for this review. Comprehensive details of the individual trials are provided in tabular form, and their results, grouped as indicated above, have been presented in text and analyses tables. The poor quality and heterogeneity in terms of patient characteristics, interventions compared and outcome measurement, of the included trials meant that no meta-analyses were undertaken. REVIEWER'S CONCLUSIONS: There remains insufficient evidence from randomised trials to determine which methods of conservative treatment are the most appropriate for the more common types of distal radial fractures in adults. Therefore, at present, practitioners applying conservative management should use an accepted technique with which they are familiar, and which is cost-effective from the perspective of their provider unit. Patient preferences and circumstances, and the risk of complications should also be considered. Further research to clarify the most appropriate conservative treatment for this common fracture is warranted but requires careful preparation with an emphasis on undertaking a systematic approach. For this, it is important to reflect on the issues raised in this review, to ascertain patient preferences, prioritise researchable questions and agree a core data set for classification of fractures and outcomes. PMID- 12076392 TI - Extramedullary fixation implants for extracapsular hip fractures. AB - BACKGROUND: Extramedullary fixation of hip fractures refers to the application of a plate and screws to the lateral side of the proximal femur. OBJECTIVES: To compare different types of extramedullary fixation implants for the surgical treatment of extracapsular hip fracture in adults. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register and reference lists of relevant articles. Date of the most recent search: October 2001. SELECTION CRITERIA: All randomised or quasi-randomised trials comparing extramedullary implants used in the fixation of extracapsular hip fracture in adults. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, using a ten item scale, and extracted data. Additional information was sought from all trialists. Wherever appropriate and possible, data from comparable trials were pooled. MAIN RESULTS: Two new trials investigating the Medoff plate are included in this update. All ten included trials (involving 1856 participants) had methodological flaws which may have resulted in serious bias. Three trials involving 355 patients compared a fixed nail plate (Jewett or McLaughlin) with the sliding hip screw (SHS). The limited data presented indicated an increased risk of fixation failure outcomes for fixed nail plates. The two trials, involving 433 patients, comparing the RAB plate with the SHS had contrasting results, notably in terms of operative complications, fixation failure and anatomical restoration. One trial involving 100 patients compared the Pugh nail and the SHS. There was no significant difference between implants for the outcome measures reported. Two trials involving 292 patients with 296 fractures, compared the Medoff plate with the SHS. Higher mean operative blood loss and longer mean operative time were reported for the Medoff plate in one trial. There was a lower risk of fixation failure for unstable trochanteric fractures fixed with the Medoff plate. Two trials involving, respectively, 569 patients with unstable trochanteric fractures and 107 with subtrochanteric fractures compared the Medoff plate with three different screw-plate systems. There were no statistically significant differences in outcome for trochanteric fractures. For subtrochanteric fractures, there was a tendency to a lower fixation failure rate for the Medoff plate, but no evidence for differences in longer-term outcomes. REVIEWER'S CONCLUSIONS: The fixed nail plate has higher risks of implant breakage and fixation failure than the SHS. Though insufficient evidence on other outcomes is available from randomised trials, the increased fixation failure rate is a major consideration and thus the SHS appears preferable. Insufficient information is available to draw firm conclusions of the clinical significance of differences between the SHS and either the RAB plate, the Pugh nail or the Medoff plate. PMID- 12076393 TI - Sequential combination of glucocorticosteroids and alfa interferon versus alfa interferon alone for HBeAg-positive chronic hepatitis B. AB - BACKGROUND: Chronic hepatitis B has serious effects on morbidity and mortality. Alfa interferon has been shown to increase the rates of HBeAg-clearance as well as seroconversion to anti-HBe, but response rates are unsatisfactory. Glucocorticosteroid pretreatment may increase the response to alfa interferon. OBJECTIVES: The objectives were to assess the effects of the sequential combination of glucocorticosteroids and alfa interferon versus alfa interferon alone in hepatitis B 'e' antigen positive chronic hepatitis B on mortality, virological response, biochemical response, liver histology, quality of life, and adverse events. SEARCH STRATEGY: Electronic searches of the controlled trial registers of The Cochrane Hepato-Biliary Group and The Cochrane Library, MEDLINE, BIOSIS, and EMBASE were combined (May 2000). Reading the bibliography of retrieved articles identified further trials. Alfa interferon-manufacturing companies were approached in order to inquire about any published and unpublished randomised trials. SELECTION CRITERIA: The analyses included randomised trials comparing identical alfa interferon treatment regimens with and without glucocorticosteroid pretreatment for hepatitis B 'e' antigen positive chronic hepatitis. The trials could be open, single blinded, or double blinded. No patient exclusion criteria were applied. DATA COLLECTION AND ANALYSIS: Three reviewers independently selected the trials and one extracted the data, which were validated. Assessments of the outcome measures were performed at the end of treatment and at six months and at maximal follow up after the end of treatment with alfa interferon. MAIN RESULTS: A total of 13 randomised trials including 790 patients were included. Loss of hepatitis B 'e' antigen (OR 1.41, 95% confidence interval 1.03 to 1.92, P = 0.03) and hepatitis B virus DNA (OR = 1.51, 95% confidence interval 1.12 to 2.05, P = 0.008) were significantly more frequent among patients treated with the sequential combination of glucocorticosteroids and alfa interferon than among patients treated with alfa interferon alone. Glucocorticosteroid pretreatment did not significantly influence seroconversion from hepatitis B 'e' antigen to antibodies to hepatitis B 'e' antigen, loss of hepatitis B surface antigen, normalisation of alanine aminotransferase/aspartate aminotransferase activities, and severity of adverse events. Glucocorticosteroid pretreatment did not significantly affect mortality and adverse events. The effect of glucocorticosteroid pretreatment on liver histology and quality of life could not be assessed due to insufficient data. REVIEWER'S CONCLUSIONS: Pretreatment with glucocorticosteroids before treatment with alfa interferon in patients with hepatitis B 'e' antigen positive chronic hepatitis B may be more effective than treatment with alfa interferon alone with regard to loss of hepatitis B 'e' antigen and hepatitis B virus DNA, but evidence for effect on clinical outcomes is lacking. PMID- 12076394 TI - Interferon for interferon naive patients with chronic hepatitis C. AB - BACKGROUND: A previous meta-analysis of interferon therapy in naive patients with chronic hepatitis C has documented its efficacy in achieving virologic clearance, and improving liver biochemistry and histology; however, since its publication additional trials have been reported. OBJECTIVES: To evaluate the response to interferon in interferon naive patients with chronic hepatitis C. The effect of treatment dose and duration, and the response in patients with cirrhosis and those with normal aminotransferases was also investigated. SEARCH STRATEGY: The Cochrane Controlled Trials Register (Cochrane Library Issue 1, 1999), MEDLINE (January 1966 to December 1999), and reference lists were searched, and pharmaceutical companies were contacted for unpublished trials. SELECTION CRITERIA: Randomised clinical trials comparing interferon with placebo, no treatment, or different regimens of interferon were selected. Abstracts were excluded. DATA COLLECTION AND ANALYSIS: The primary outcome measure was sustained disappearance of serum HCV RNA (virologic sustained response (SR)). Biochemical and end of treatment responses, liver histology, and adverse events were also recorded. Assessment of drug efficacy used the methods of Peto and Der Simonian and Laird. MAIN RESULTS: Fifty-four trials enrolling 6545 patients were included. Compared with no treatment, interferon 3 MU thrice weekly for 12 months increased the probability of a virologic SR (Peto odds ratio (OR) 4.60; 95% confidence interval (CI) 1.53 to 13.85). At this dosage and duration of therapy, the rate of virologic SR was 17% (95% CI 10 to 28%) in interferon-treated patients versus 3% (95% CI 1 to 10%) in controls. A dose of 6 MU was more effective than 3 MU thrice weekly (OR for 12 months treatment, 2.21; 95% CI 1.10 to 4.45), as were durations of 12 months or greater versus six months (OR 1.87; 95% CI 1.30 to 2.67). Adverse events were more common with higher doses and prolonged durations of treatment. Compared with no therapy, interferon increased the probability of histologic improvement (OR 9.22; 95% CI 5.69 to 14.94). The response to interferon in cirrhotic patients (virologic SR, 17%; 95% CI 11 to 26%) was similar to that in non-cirrhotic patients. However, interferon was no more effective than control in patients with normal aminotransferases. REVIEWER'S CONCLUSIONS: Interferon is effective in achieving viral clearance and improving liver biochemistry and histology in interferon naive patients with chronic hepatitis C. Higher doses and prolonged durations are more effective, but associated with more frequent adverse events. Interferon is associated with similar benefits in patients with cirrhosis, but the efficacy in patients with normal aminotransferases is unproven. PMID- 12076395 TI - Kinesthetic stimulation for preventing apnea in preterm infants. AB - BACKGROUND: Recurrent apnea is common in preterm infants, particularly at very early gestational ages. These episodes of loss of effective breathing can lead to hypoxemia and bradycardia, which may be severe enough to require resuscitation including use of positive pressure ventilation or other treatments. Physical stimulation is often used to restart breathing and it is possible that repeated stimulation, such as with an oscillating mattress (kinesthetic stimulation), might prevent apnea and its consequences. OBJECTIVES: In preterm infants at risk for apnea, does prophylactic use of kinesthetic stimulation lead to a clinically important reduction in apnea and bradycardia, and use of intermittent positive pressure ventilation (IPPV). SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group was used. This included searches of the Oxford Database of Perinatal trials, Cochrane Controlled Trials Register (Cochrane Library, Issue 4, 2001), MEDLINE (1966-January 2002), EMBASE, CINAHL (1982-January 2002), previous reviews including cross references, abstracts, conferences and symposia proceedings, expert informants, journal hand searching mainly in the English language. SELECTION CRITERIA: All trials in preterm infants at risk of developing clinical apnea which utilized random or quasi-random allocation to treatment with an oscillating mattress (or other forms of repetitive kinesthetic stimulation) or control, were eligible. DATA COLLECTION AND ANALYSIS: Standard methods of the Cochrane Collaboration and its Neonatal Review Group were used with separate evaluation of trial quality and data extraction by each author and synthesis of the data using relative risk. MAIN RESULTS: Three trials enrolling a total of 154 babies were included in this review. There was no evidence of effect on short term outcomes (apnea /bradycardia, IVH, use of IPPV, sleep/wake cycles and neurological status at discharge) or long term outcomes (in one trial - growth and development to one year). REVIEWER'S CONCLUSIONS: Implications for practice. Prophylactic use of kinesthetic stimulation cannot be recommended to reduce apnea/bradycardia in preterm infants. Implications for research. There are currently no clear research questions regarding prophylactic use of kinesthetic stimulation to prevent apnea in preterm infants. PMID- 12076396 TI - Interventions for treating proximal humeral fractures in adults. AB - BACKGROUND: Proximal humeral fractures are common yet management varies widely. In particular, the role and timing of any surgical intervention have not been clearly defined. OBJECTIVES: To collate and evaluate the scientific evidence supporting the various methods used for treating proximal humeral fractures. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register, MEDLINE, EMBASE, the Cochrane Controlled Trials Register, CINAHL, the National Research Register (UK), AMED, PEDro and bibliographies of trial reports. The search was completed in November 2001. SELECTION CRITERIA: All randomised studies pertinent to the treatment of proximal humeral fractures were selected. DATA COLLECTION AND ANALYSIS: Independent quality assessment and data extraction were performed by two reviewers. Although quantitative data from trials are presented, trial heterogeneity prevented pooling of results. MAIN RESULTS: Ten randomised trials were included. All were small trials; the largest study involved only 85 patients. Bias in these trials could not be ruled out. Six trials evaluated conservative treatment, three compared surgery with conservative treatment and one compared two surgical techniques. In the 'conservative' group there was very limited evidence indicating that the type of bandage used made any difference in terms of time to fracture union and the functional end result. However, an arm sling was generally more comfortable than a body bandage. There was some evidence that mobilisation at one week instead of three weeks alleviated pain in the short term without compromising long term outcome. Two trials provided some evidence that patients, when given sufficient instruction to pursue an adequate physiotherapy programme, could generally achieve a satisfactory outcome if allowed to exercise without supervision. Operative reduction improved fracture alignment in two trials. However, in one trial, surgery was associated with a greater risk of complication, and did not result in improved shoulder function. In one trial, hemi-arthroplasty resulted in better short-term function with less pain and less need for help with activities of daily living when compared with conservative treatment for severe injuries. Fracture fixation of severe injuries was associated with a high rate of re-operation in one trial, comparing tension-band wiring fixation with hemi-arthroplasty. REVIEWER'S CONCLUSIONS: Only tentative conclusions can be drawn from the available randomised trials, which do not provide robust evidence for many of the decisions that need to be made in contemporary fracture management. It is unclear whether operative intervention, even for specific fracture types, will produce consistently better long term outcomes. There is a need for good quality evidence for the management of these fractures. PMID- 12076397 TI - Radiant warmers versus incubators for regulating body temperature in newborn infants. AB - BACKGROUND: The provision of a thermoneutral environment is an essential component of the immediate and longer term care of newborn infants. A variety of methods are currently employed including incubators and open-care systems, with or without modifications such as heat shields and plastic wrap. The system used must allow ready access to the infant but should also minimise alterations in the immediate environment. OBJECTIVES: To assess the effects of radiant warmers versus incubators on neonatal fluid and electrolyte balance, morbidity and mortality. SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Review Group was used. This included searches of electronic databases: Oxford Database of Perinatal Trials, Cochrane Controlled Trials Register (Cochrane Library Issue 4 2001), MEDLINE (1966 -2001), and CINAHL (1982-2001). SELECTION CRITERIA: Randomised or quasi-randomised trials in which radiant warmers were compared to incubators in a neonatal population. DATA COLLECTION AND ANALYSIS: Independent data extraction and quality assessment of included trials was conducted by the authors. Data were analysed using relative risk (RR) and weighted mean difference (WMD). Results are presented with 95% confidence intervals. Meta-analysis was undertaken using a fixed effect model. MAIN RESULTS: Eight studies are included in this review; six employed a crossover design. In the overall comparison of radiant warmers vs incubators, radiant warmers caused a statistically significant increase in insensible water loss (IWL) [WMD 0.94g/Kg/day (95% CI 0.47, 1.41)] and a trend towards increased oxygen consumption which was not statistically significant [WMD 0.27mL/kg/min (95% CI 0.09, 0.63)]. Due to small numbers, effects on important clinical outcomes could not be adequately assessed. A comparison of radiant warmers with heat shields vs incubators without heat shields showed a trend for increased IWL in the radiant warmer group, which was not statistically significant. No difference was shown in oxygen consumption. REVIEWER'S CONCLUSIONS: Radiant warmers result in increased IWL compared to incubators. This needs to be taken into account when calculating daily fluid requirements. The results of this review do not provide sufficient evidence concerning effects on important outcomes to guide clinical practice. Further randomised controlled trials are required to assess the effects of radiant warmers versus incubators in neonatal care on important short and long term outcomes, with particular attention to extremely low birthweight infants in the early neonatal period. PMID- 12076398 TI - Supported housing for people with severe mental disorders. AB - BACKGROUND: There has been a significant reduction in the number of people with severe mental illness who spend extended periods in long-stay hospitals. Psychiatric and social services, both statutory and voluntary, aim to assist these people to stay in their local community. District health authorities, local authorities, housing associations and voluntary organisations are jointly expected to provide support for people with severe mental disorder/s. This 'support' may well involve some sort of special housing. OBJECTIVES: To determine the effects of supported housing schemes compared with outreach support schemes or 'standard care' for people with severe mental disorder/s living in the community. SEARCH STRATEGY: Cochrane Schizophrenia Group's Register of trials (February 2001) and the Cochrane Library (Issue 1, 2001) were searched using relevant phrases. These databases are compiled by methodical searches of BIOSIS, CINAHL, Dissertation abstracts, EMBASE, LILACS, MEDLINE, PSYNDEX, PsycINFO, RUSSMED, Sociofile, supplemented with hand searching of relevant journals and numerous conference proceedings. Reference list screening of relevant papers was performed. SELECTION CRITERIA: Relevant randomised, or quasi-randomised, trials dealing with people with 'severe mental disorder/s' allocated to supported housing, outreach support schemes or standard care focusing on outcomes of service utilisation, mental state, satisfaction with care, social functioning, quality of life, and economic data, were sought. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality rated and data extracted. For dichotomous data, relative risks (RR) would have been estimated, with the 95% confidence intervals (CI). Where possible, the number needed to treat statistic (NNT) was to have been calculated. Analysis would have been by intention-to-treat. Normal continuous data were to have been summated using the weighted mean difference (WMD). Scale data were to have been presented for only those tools that had attained pre-specified levels of quality. Tests of heterogeneity and for publication bias were to have been undertaken. MAIN RESULTS: No studies met the inclusion criteria although 139 citations were acquired from the searches. REVIEWER'S CONCLUSIONS: Dedicated schemes whereby people with severe mental illness are located within one site or building with assistance from professional workers have potential for great benefit as they provide a 'safe haven' for people in need of stability and support. This, however, may be at the risk of increasing dependence on professionals and prolonging exclusion from the community. Whether or not the benefits outweigh the risks can only be a matter of opinion in the absence of reliable evidence. There is an urgent need to investigate the effects of supported housing on people with severe mental illness within a randomised trial. PMID- 12076399 TI - Psychological debriefing for preventing post traumatic stress disorder (PTSD). AB - BACKGROUND: Over approximately the last last fifteen years early psychological interventions such as psychological 'debriefing' have been increasingly used to treat psychological trauma. While these intervention have become popular and their use spread to several settings - efficacy had largely not been tested empirically. In 1997 a systematic review of single session psychological "debriefing" was undertaken and this subsequently became a protocol and Cochrane Review published in 1998 (Issue2). This update forms the first substantive update of the original review. OBJECTIVES: To assess the effectiveness of brief psychological debriefing for the management of psychological distress after trauma, and the prevention of post traumatic stress disorder. SEARCH STRATEGY: Electronic searching of MEDLINE, EMBASE, PsychLit, PILOTS, Biosis, Pascal, Occ. Safety and Health,SOCIOFILE, CINAHL, PSYCINFO, PSYNDEX, SIGLE, LILACS, CCTR, CINAHL, NRR, Hand search of Journal of Traumatic Stress. Contact with leading researchers. SELECTION CRITERIA: The inclusion criteria for all randomized studies was that they should focus on persons recently (one month or less) exposed to a traumatic event, should consist of a single session only, and that the intervention involve some form of emotional processing/ventilation by encouraging recollection/reworking of the traumatic event accompanied by normalisation of emotional reaction to the event. DATA COLLECTION AND ANALYSIS: 11 trials fulfilled the inclusion criteria. Quality was generally poor. Data from two trials could not be synthesised. Two trials involved the use of the intervention in an obstetric setting. MAIN RESULTS: Single session individual debriefing did not reduce psychological distress nor prevent the onset of post traumatic stress disorder (PTSD). Those who received the intervention showed no significant short term (3-5 months) in the risk of PTSD (odds ratio 1.22 (95% ci 0.60 to 2.46 )). At one year one trial reported that there was a significantly increased risk of PTSD in those receiving debriefing (odds ratio 2.88 (1.11 to 7.53))odds ratio 95%). There was also no evidence that debriefing reduced general psychological morbidity, depression or anxiety. REVIEWER'S CONCLUSIONS: There is no current evidence that psychological debriefing is a useful treatment for the prevention of post traumatic stress disorder after traumatic incidents. Compulsory debriefing of victims of trauma should cease. PMID- 12076400 TI - Limited (information only) patient education programs for adults with asthma. AB - BACKGROUND: A key component of many asthma management guidelines is the recommendation for patient education and regular medical review. A number of controlled trials have been conducted to measure the effectiveness of asthma education programmes. These programmes improve patient knowledge, but their impact on health outcomes is less well established. At its simplest level, education is limited to the transfer of information about asthma, its causes and its treatment. This review focused on the effects of limited asthma education. OBJECTIVES: The objective of this review was to assess the effects of limited (i.e. information only) asthma education on health outcomes in adults with asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles. SELECTION CRITERIA: Randomised and controlled trials of individual asthma education involving information transfer only in adults over 16 years of age. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and two reviewers extracted data independently. Study authors were contacted for missing information. MAIN RESULTS: Twelve trials were included. They were of variable quality. Limited asthma education did not reduce hospitalisation for asthma (weighted mean difference -0.03 average hospitalisations per person per year, 95% confidence interval -0.09 to 0.03). There was no significant effect on doctor visits, lung function and medication use. The effects on asthma symptoms were variable. There was no reduction in days lost from normal activity, but in two studies, perceived asthma symptoms did improve after limited asthma education (odds ratio 0.44, 95% confidence interval 0.26 to 0.74). In one study, limited asthma education was associated with reduced emergency department visits (reduction of -2.76 average visits per person per year, 95% confidence interval -4.34 to 1.18). REVIEWER'S CONCLUSIONS: Use of limited asthma education as it has been practiced does not appear to improve health outcomes in adults with asthma although perceived symptoms may improve. Provision of information in the emergency department may be effective, but this needs to be confirmed. PMID- 12076401 TI - Danazol for heavy menstrual bleeding. AB - BACKGROUND: Heavy menstrual bleeding (HMB) is an important cause of ill health in pre menopausal women. Medical therapy, with the avoidance of possibly unnecessary surgery is an attractive treatment option, but there is considerable variation in practice and uncertainty about the most effective therapy. Danazol is a synthetic steroid with anti-oestrogenic and anti progestogenic activity, and weak androgenic properties. Danazol suppresses oestrogen and progesterone receptors in the endometrium, leading to endometrial atrophy (thinning of the lining of the uterus) and reduced menstrual loss and to amenorrhoea in some women. OBJECTIVES: To determine the effectiveness and tolerability of danazol when used for heavy menstrual bleeding in women of reproductive years. SEARCH STRATEGY: All studies which might describe randomised controlled trials of danazol for the treatment of heavy menstrual bleeding were obtained by electronic searches of MEDLINE, EMBASE, Current Contents, CINAHL, National Research Register and the Menstrual Disorders and Subfertility Group's Specialist Register of controlled trials (on 6 November 2001). Attempts were also made to identify trials from citation lists of included trials and relevant review articles. In most cases the first author of each included trial was contacted for unpublished additional information. SELECTION CRITERIA: Randomised controlled trials of danazol versus placebo, any other medical (non-surgical) therapy or danazol in different dosages for heavy menstrual bleeding in women of reproductive age with regular HMB measured either subjectively or objectively. Trials that included women with post menopausal bleeding, intermenstrual bleeding and pathological causes of heavy menstrual bleeding were excluded. DATA COLLECTION AND ANALYSIS: Nine RCTs, with 353 women, were identified that fulfilled the inclusion criteria for this review. Quality assessment and data extraction were performed independently by two reviewers. The main outcomes were menstrual blood loss, the number of women experiencing adverse effects, weight gain, withdrawals due to adverse effects and dysmenorrhoea. If data could not be extracted in a form suitable for meta-analysis, they were presented in a descriptive format. MAIN RESULTS: Most data were not in a form suitable for meta analysis, and the results are based on a small number of trials, all of which are under-powered. Danazol appears to be more effective than placebo, progestogens, NSAIDs and the OCP at reducing MBL, but confidence intervals were wide. Treatment with danazol caused more adverse events than NSAIDs (OR 7.0; 95% CI 1.7, 28.2) and progestogens (OR 4.05, 95% CI 1.6, 10.2), but this did not appear to affect adherence to treatment. Danazol was shown to significantly lower the duration of menses when compared with NSAIDs (WMD -1.0; 95% CI -1.8, -0.3) and a progesterone releasing IUD (WMD -6.0; 95% CI -7.3, 4.8). There were no randomised trials comparing danazol with tranexamic acid or the levonorgestrel-releasing intrauterine system. REVIEWER'S CONCLUSIONS: Danazol appears to be an effective treatment for heavy menstrual bleeding compared to other medical treatments, though it is uncertain whether it is acceptable to women. The use of danazol may be limited by its side effect profile, its acceptability to women and the need for continuing treatment. Overall no strong recommendations can be made due to the small number of trials, and the small sample sizes of the included trials. PMID- 12076402 TI - Continuous positive airways pressure for obstructive sleep apnoea. AB - BACKGROUND: Obstructive sleep apnoea is the periodic reduction (hypopnoea) or cessation (apnoea) of breathing due to narrowing or occlusion of the upper airway during sleep. The main symptom is daytime sleepiness although there it has been linked to premature death, hypertension, ischaemic heart disease, stroke and road traffic accidents. OBJECTIVES: The main treatment for sleep apnoea is with continuous positive airways pressure (CPAP) treatment, which consists of a flow generator and mask. These are used at night to prevent apnoea, hypoxia and sleep disturbance. The objective was to assess the effects of CPAP in the treatment of obstructive sleep apnoea in adults. SEARCH STRATEGY: We searched the Cochrane Airways Group RCT register (MEDLINE 1966 to 2000, Embase 1974 to 2000, Cinahl 1982 to 2000) and the reference lists of articles. We consulted experts in the field. SELECTION CRITERIA: Randomised trials comparing nocturnal CPAP with placebo or other treatments in adults with obstructive sleep apnoea and an apnoea/hypopnoea index greater than five per hour. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and two reviewers extracted data independently. Study authors were contacted for missing information. MAIN RESULTS: Twelve trials involving 475 people were included. Most studies had methodological shortcomings. Most trials were of crossover design. Compared with placebo, CPAP showed significant improvements in objective and subjective sleepiness and several quality of life and depression measures. Patients preferred CPAP to placebo (odds ratio 0.4, 95% confidence interval 0.2 to 0.8). There was no significant effect on daytime blood pressure. Compared with oral appliances, CPAP significantly improved the apnoea/hypopnoea index (weighted mean difference -7.3, 95% confidence interval -10.0 to -4.7) and minimum oxygen saturation during sleep. Patients strongly preferred the oral appliance to CPAP (odds ratio 9.5, 95% confidence interval 4.3 to 21.1). REVIEWER'S CONCLUSIONS: CPAP is more effective than placebo in improving sleepiness and quality of life measures for people with obstructive sleep apnoea. It is more effective than oral appliances in improving respiratory disturbances. Although patients prefer CPAP to placebo, they preference oral appliances to CPAP. PMID- 12076403 TI - Ambulatory versus conventional methods for monitoring blood pressure during pregnancy. AB - BACKGROUND: Hypertensive disorders are among the most common medical complications of pregnancy and a leading cause of maternal and perinatal morbidity and mortality world-wide. Blood pressure measurement plays a central role in the screening and management of hypertension during pregnancy. In recent years the validity of conventional (clinic) blood pressure measurement has been questioned and efforts have been made to improve the technique with ambulatory automated devices that provide a large number of measurements over a period of time, usually a 24-hour period. OBJECTIVES: To assess whether the use of ambulatory blood pressure monitoring during pregnancy improves subsequent maternal and feto-neonatal outcomes, women-newborn quality of life or use of health service resources, compared with conventional (clinic) blood pressure measurements. These effects will be assessed for the following subgroups: (1) Women at low or average risk of hypertensive disorders of pregnancy (unselected). (2) Women defined as high risk of hypertensive disorders of pregnancy. (3) Women with hypertension without other signs of pre-eclampsia. (4) Women with established pre-eclampsia. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register, MEDLINE, LILACS and EMBASE were searched. Date of last search: July 2001. SELECTION CRITERIA: All randomised trials comparing ambulatory blood pressure monitoring versus conventional (clinic) blood pressure monitoring in pregnancy. Quasi-random designs will be excluded. DATA COLLECTION AND ANALYSIS: Two reviewers evaluated all potentially relevant articles, examined each study for possible inclusion and assessed the methodological quality using the Cochrane guidelines. MAIN RESULTS: No trials included. REVIEWER'S CONCLUSIONS: There is no randomised controlled trial evidence to support the use of ambulatory blood pressure monitoring during pregnancy. Randomized trials with adequate design and sample sizes are needed to evaluate the possible advantages and risks of ambulatory blood pressure monitoring during pregnancy, in particular in hypertensive pregnant women. These trials should evaluate not only clinical outcomes, but also use of health care resources and women's views. PMID- 12076404 TI - Intra-venous albumin for preventing severe ovarian hyperstimulation syndrome. AB - BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is an iatrogenic condition that occurs after the administration of human menopausal gonadotrophin (hMG) with or without gonadotrophin releasing hormone (GnRH) agonists. OHSS is a threat to every woman undergoing ovulation induction and is potentially lethal in its severest form. Severe OHSS is characterised by growth of multiple large follicles with massive extravascular protein rich fluid shift. This may lead to hypovolaemia, haemoconcentration, oliguria, and electrolyte disturbance. Human albumin solutions are now used in the management of shock and other conditions in which restoration of blood volume is urgent, the acute management of burns, and clinical situations associated with hypoproteinaemia. Recently, a number of clinical trials with conflicting results have been reported in which albumin has been tested as a possible way for preventing the severe form of OHSS. OBJECTIVES: To review the effectiveness of human albumin administration in prevention of severe ovarian hyperstimulation syndrome. SEARCH STRATEGY: The Menstrual Disorders and Subfertility Group literature search strategy was used to identify randomised trials that had compared the use of human albumin with placebo or no treatment in the prevention of severe ovarian hyperstimulation syndrome. A diverse search strategy was employed, including handsearching of core journals from 1966 to November 2001, searching bibliographies of relevant trials, MEDLINE, EMBASE, PsychLIT and CINAHL databases, the MDSG specialised register, abstracts from North American and European meetings and contact with authors of relevant papers. SELECTION CRITERIA: Trials were included if they compared the effect of human albumin with placebo or no treatment on relevant outcomes. Only randomised controlled studies were included in this review. DATA COLLECTION AND ANALYSIS: Seven randomised controlled trials were identified, five of which met our inclusion criteria and enrolled 378 women (193 in the albumin treated group and 185 in the control group). Trials under consideration were evaluated for methodological quality and appropriateness for inclusion without consideration of their results.The five included trials were single-centre parallel randomised controlled studies. Relevant data were extracted independently by two reviewers using the standardized data extraction sheet. Validity was assessed in terms of method of randomisation, completeness of follow-up, presence or absence of crossover and co-intervention. DATA SYNTHESIS: 2x2 tables were generated for all relevant outcomes. Odds ratios were calculated using the Peto modified Mantel Haenszel technique. MAIN RESULTS: Meta-analysis of the five included trials demonstrated significant reduction in severe ovarian hyperstimulation syndrome on administration of human albumin (odds ratio was 0.28 (95% CI 0.11 to 0.73). Relative risk was 0.35 (0.14 - 0.87) and absolute risk reduction was 5.5. For every 18 women at risk of severe OHSS, albumin infusion will save one more case. There was no evidence of an increase in the pregnancy rate (odds ratio was 1.09, (95% CI 0.65 to 1.83) REVIEWER'S CONCLUSIONS: This review shows a clear benefit from administration of intra-venous albumin at the time of oocyte retrieval in prevention of severe OHSS in high-risk cases. Whether the NNT would justify the routine use of albumin infusion in cases at risk of severe OHSS needs to be judged by clinical decision makers. PMID- 12076405 TI - Surgical interventions for age-related cataract. AB - BACKGROUND: Cataract is the major cause of global blindness, accounting for 40 to 80% of all blindness in developing countries. The number of people blind from cataract is expected to rise due to the changing age distribution and increasing life expectancy. There is currently no proven intervention to prevent cataract and surgery is the only form of treatment. OBJECTIVES: The objective of this review is to compare the effects of different surgical interventions for age related cataract. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register - CENTRAL/CCTR, which contains the Cochrane Eyes and Vision Group specialised register (Cochrane Library Issue 3 2001), MEDLINE (1966 to August 2001), EMBASE (1980 to September 2001), the reference lists of identified trials, and we contacted investigators and experts in the field for details of published and unpublished trials. SELECTION CRITERIA: We included randomised controlled trials evaluating surgical treatment for people with age-related cataract. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and discrepancies were resolved by discussion. Where appropriate, relative risks, odds ratios and weighted mean differences were summarised after assessing heterogeneity between the studies. We used a fixed effect model due to the low number of trials in each comparison. MAIN RESULTS: We identified six trials that randomised a total of 7828 people. Phacoemulsification gave a better visual outcome than extracapsular surgery and gave a similar average cost per procedure in one trial conducted in the UK. Extracapsular surgery with posterior chamber lens implant and intracapsular surgery with or without an anterior chamber intraocular lens implant gave acceptable visual outcomes at 12 to 24 months after surgery. In three large trials in south Asia, best-corrected visual acuity of less than 6/60 ranged from 0.5 to 4%. Higher rates of poor outcome were observed in a multicentre study with 19 surgeons compared to a single-centre study with two surgeons. REVIEWER'S CONCLUSIONS: This review provides evidence from one randomised controlled trial that phacoemulsification gives a better visual outcome than extracapsular extraction with sutures. However, this trial was conducted in a developed country specialised hospital setting and extrapolation to other settings must be made with caution. This review also found evidence that extracapsular cataract extraction with a posterior chamber lens implant provides better visual outcome than intracapsular extraction with aphakic glasses. This finding is also based on the results of a single trial. The long term effects of posterior capsular opacification need to be assessed in larger populations. The data in the review suggest that intracapsular extraction with an anterior chamber lens implant is an effective alternative to intracapsular extraction with aphakic glasses, with similar safety. Further data from developing regions are needed to compare all aspects of intraocular lens surgery with the three main surgical procedures - intracapsular extraction with an anterior chamber lens, extracapsular surgery with a posterior chamber lens with or without sutures. PMID- 12076406 TI - Naltrexone maintenance treatment for opioid dependence. AB - BACKGROUND: Despite widespread use of naltrexone maintenance in many countries for more than a decade, the evidence of its effects has not yet been systematically evaluated. OBJECTIVES: To evaluate the effects of naltrexone maintenance treatment in preventing relapse in opioid addicts after detoxification. SEARCH STRATEGY: We searched MEDLINE (1973-first year of naltrexone use in humans-July 2000), EMBASE (1974-July 2000), Cochrane Controlled Trials Register (Cochrane Library issue 2001.4) and handsearched the "Bolletino per le Farmacodipendenze e l'Alcolismo" (1978 to 1997) and reference lists of relevant articles. We contacted pharmaceutical producers of naltrexone, authors and other Cochrane review groups. Date of most recent searches: December 2001. SELECTION CRITERIA: All controlled studies of naltrexone; treatment of heroin addicts after detoxification. DATA COLLECTION AND ANALYSIS: Reviewers evaluated data independently and analysed outcome measures taking into consideration adherence to and success of the study intervention. Data were extracted and analysed stratifying for the three categories of study quality. Where possible, meta-analysis was performed. MAIN RESULTS: Eleven studies met the criteria for inclusion in this review, even if not all of them were randomised. The methodological quality of the included studies varied, but was generally poor. Meta-analysis could be performed to a very low degree only, because the studies and their outcome measures were very heterogeneous. A statistically significant reduction of (re-)incarcerations was found for patients treated with naltrexone and behaviour therapy in respect to those treated with behaviour therapy only. The other outcomes considered in the meta-analysis did not yield any significant results. Final conclusions on whether naltrexone treatment may be considered effective in maintenance therapy cannot be drawn from the clinical trials available so far. REVIEWER'S CONCLUSIONS: The available trials do not allow a final evaluation of naltrexone maintenance treatment yet. A trend in favour of treatment with naltrexone was observed for certain target groups (particularly people who are highly motivated), as has been previously described in the literature. PMID- 12076407 TI - Physiotherapist-led programmes and interventions for rehabilitation of anterior cruciate ligament, medial collateral ligament and meniscal injuries of the knee in adults. AB - BACKGROUND: Soft-tissue injuries of the knee, mainly involving the anterior cruciate ligament (ACL), the medial collateral ligament (MCL) and menisci, are common and their rehabilitation after non-surgical or surgical treatment often involves intensive and prolonged physiotherapy. OBJECTIVES: To examine the evidence for effectiveness of various physiotherapist-led (or 'directed') rehabilitation programmes, and of various interventions used within these programmes, for rehabilitation of acute or chronic ACL, MCL or meniscal injuries of the knee in adults. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group's specialised register (to June 2001), MEDLINE (from 1966 to August 1999), EMBASE (from 1980 to February 1997), CINAHL (1982 to April 1999), CURRENT CONTENTS (up to March 1999) and reference lists of relevant articles, and consulted colleagues. Date of the most recent search: June 2001. SELECTION CRITERIA: Randomised or quasi-randomised clinical trials evaluating physiotherapist-led rehabilitation programmes, or components of rehabilitation programmes, for the treatment or post-surgical rehabilitation of ACL, MCL or knee meniscal injuries. Excluded were trials investigating electrical stimulation, or various interventions such as cryotherapy, immobilisation braces and continuous passive motion when used in initial or early treatment. Laboratory based trials reporting intermediate outcomes were also excluded. DATA COLLECTION AND ANALYSIS: All trials, judged as fitting the selection criteria by two reviewers, were independently assessed by two reviewers for methodological quality by use of an 11 item checklist. Data were independently extracted by two reviewers. Any disagreement was resolved by discussion. Although quantitative data from most trials are presented, using relative risks or mean differences together with 95 per cent confidence intervals, trial heterogeneity and lack of outcome data prevented meaningful pooling of results from comparable trials. MAIN RESULTS: Thirty-one trials, involving 1545 mainly young and male patients, met the inclusion criteria of the review. Methodological quality was highly variable: allocation concealment and / or assessor blinding were rare, and assessment of outcome was often incomplete and short-term. ACL injury and /or deficiency was the main focus of 18 trials, MCL injury of two trials, meniscal injury of nine trials and a mixture of soft-tissue injuries in the other two trials. The trial comparisons fell into five main categories: rehabilitation programme versus control (6 trials); one rehabilitation programme versus another (6 trials); different timing of rehabilitation (4 trials); one component of a programme versus another (6 trials); supplementary interventions to a programme versus none (9 trials). No trial provided sufficient evidence to establish the relative effectiveness of the intervention(s) under investigation. REVIEWER'S CONCLUSIONS: The available evidence for physiotherapist-led rehabilitation of ACL, MCL and meniscal injuries is wide ranging in terms of scope but insufficient to establish the relative effectiveness of the various approaches and methods in current use. There is a need for further research involving good quality, large scale randomised trials with sufficiently long follow-up to fully assess knee function and recovery. PMID- 12076408 TI - Amisulpride for schizophrenia. AB - BACKGROUND: The treatment of schizophrenia with old, 'typical' antipsychotic drugs such as haloperidol can be problematic, because many people treated with these drugs will suffer from movement disorders. Amisulpride is said to be an "atypical" antipsychotic which induces less movement disorder and which is effective for the negative symptoms of schizophrenia. OBJECTIVES: To evaluate the effects of amisulpride as compared with placebo, typical and atypical antipsychotic drugs for schizophrenia. SEARCH STRATEGY: The authors carried out electronic searches of Biological Abstracts (1982-1999), CINAHL (1982-1999), Cochrane Library (Issue 4, 1999), Cochrane Schizophrenia Group's Register (November 2000), EMBASE (1980-1999), LILACS(1982-1999), MEDLINE (1966-1999) and PsycLIT (1974-1999). They checked all identified studies for further trial citations, and sought these studies in the Science Citation Index. They also contacted authors of trials and the manufacturer of amisulpride. SELECTION CRITERIA: All randomised controlled trials comparing amisulpride to placebo, typical or atypical antipsychotic drugs for schizophrenia or other non-affective serious mental illnesses. DATA COLLECTION AND ANALYSIS: Data were independently extracted and analysed on an intention-to-treat basis. The relative risk (RR) and 95% confidence intervals (CI) of dichotomous data were calculated using a random effects model, and, where possible, the number needed to treat was calculated. Weighted mean differences (WMD) were calculated for continuous data. MAIN RESULTS: This review currently includes 19 randomised studies with a total of 2443 participants. Most trials were of short duration. Data from 4 trials with 514 participants with predominantly negative symptoms suggest that low-dose (up to 300mg/day) amisulpride was a more acceptable treatment than placebo (n=514, RR 0.6 CI 0.5 to 0.8, NNT 3 CI 3 to 7), the improvement of the participants' global state (n=242, RR 0.6 CI 0.5 to 0.8, NNT 3 CI 2 to 6) and the treatment of negative symptoms (n=177, WMD -10.1 CI -16.6 to -3.5). Amisulpride was shown to be more likely to cause extrapyramidal symptoms than placebo in two studies (n=269, RR 2.2 CI 1.2 to 4.2), but this result did not hold calculating the risk reduction so that an NNT-statistic could not be indicated. Compared to typical antipsychotics, the pooled results of a total of fourteen trials suggest that amisulpride was more effective in improving global state (n=651, RR 0.7 CI 0.5 to 0.9, NNT 6 CI 4 to 11), the general mental state (n=695, WMD -4.2 CI -6.5 to 1.9) and the negative symptoms of schizophrenia (n=506, WMD -2.8 CI -4.3 to 1.3). Regarding positive symptoms, amisulpride was as effective as typical antipsychotics. Amisulpride was less prone to cause at least one general adverse event (n=751, RR 0.9 CI 0.8 to 0.97, NNH 9 CI 6 to 18), one extrapyramidal symptom (n=771, RR 0.7 CI 0.6 to 0.9, NNH 5 CI 4 to 9) or to require the use of antiparkinson medication (n=851, RR 0.6 CI 0.5 to 0.8, NNH 4 CI 3 to 6). No clear differences in other adverse events compared to typical drugs were found. Amisulpride also seemed to be more acceptable than conventional drugs as measured by the outcome 'leaving the studies early' (n=1512, RR 0.8 CI 0.7 to 0.9, NNT 16 CI 9 to 69) than conventional drugs, but this result might have been overestimated due to a publication bias which could not be excluded with certainty. A single trial compared amisulpride to another 'atypical' antipsychotic, risperidone. With the exception of agitation, which was more frequent in the amisulpride group (n=228, RR 3.4 CI 1.2 to 10.1, NNH 11 CI 6 to 50) no significant differences were recorded on efficacy or acceptability. REVIEWER'S CONCLUSIONS: This systematic review confirms that amisulpride is an effective 'atypical' antipsychotic drug for those with schizophrenia. Amisulpride may offer a good general profile, at least compared to high-potency 'typical' antipsychotics. It may also yield better results in some specific outcomes related to efficacy, such as improvement of global state and general negative symptoms. It might be more acceptable and more tolerable than high-potency conventional antipsychotics, especially regarding extrapyramidal side-effects. Longer term randomised trials are needed to evaluate the comparative value of amisulpride, particularly compared to other expensive atypical antipsychotics. These should focus on important outcomes which have not been sufficiently monitored such as service use, family burden and quality of life. PMID- 12076409 TI - Zonisamide add-on for drug-resistant partial epilepsy. AB - BACKGROUND: The majority of people with epilepsy have a good prognosis and their seizures can be well controlled with the use of a single antiepileptic agent, but up to 30 per cent develop refractory epilepsy, especially those with partial seizures. In this review we summarize the current evidence regarding zonisamide, when used as an add-on treatment for drug-resistant partial epilepsy. OBJECTIVES: To evaluate the effects of zonisamide when used as an add-on treatment for people with drug -resistant partial epilepsy. SEARCH STRATEGY: We searched the Cochrane Epilepsy Group trial register (14/12/01), the Cochrane Controlled Trials Register (Cochrane Library Issue 4, 2001). In addition, we contacted Dainippon and Elan Pharma (makers and licensees of zonisamide) and experts in the field to seek any ongoing studies or unpublished studies. SELECTION CRITERIA: Randomized placebo controlled add-on trials of zonisamide in people with drug-resistant partial epilepsy. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion and extracted relevant data. Outcomes were: (a) fifty per cent or greater reduction in total seizure frequency; (b) treatment withdrawal (any reason); (c) adverse events. Primary analyses were intention to treat. Summary odds ratios (ORs) were estimated for each outcome. MAIN RESULTS: Three trials (499 participants) were included. The overall odds ratio (OR, 95% Confidence Interval (CI)) for 50 per cent reduction in seizure frequency compared to placebo was 2.07(1.36 to 3.15) for a 400mg/day dose of zonisamide. When the full treatment period of 12 weeks was considered for all three trials including varied rates of titration to 400mg/day the OR compared to placebo was 2.72(95% CI 1.74 to 4.25). There was insufficient evidence to support a dose response relationship for this outcome. The OR for treatment withdrawal was 1.74(95% CI 1.03 to 2.95). The 99% CI for the following side effects indicate that they are significantly associated with zonisamide: ataxia 3.94(1.23 to 12.57); somnolence 2.11(1.11 to 3.98); agitation 3.52(1.26 to 9.68); agitation and irritability 2.43(1.04 to 5.66) and anorexia 2.98(1.38 to 6.42). REVIEWER'S CONCLUSIONS: Zonisamide has efficacy as an add-on treatment in people with drug-resistant partial epilepsy. Minimum effective and maximum tolerated doses cannot be identified. The trials reviewed were of 12 week duration and results cannot be used to confirm longer periods of effectiveness in seizure control. The results cannot be extrapolated to monotherapy or to people with other seizure types or epilepsy syndromes. PMID- 12076410 TI - Sensory stimulation for brain injured individuals in coma or vegetative state. AB - BACKGROUND: Coma and vegetative state follow traumatic brain injury in about one out of eight patients, and in patients with non traumatic injury the prognosis is worse. The use of sensory stimulation for coma and vegetative state has gained popularity during the 1980's but beliefs and opinions about its effectiveness vary substantially among health professionals. OBJECTIVES: To assess the effectiveness of sensory stimulation programmes in patients in coma or vegetative state. SEARCH STRATEGY: We searched the Injuries Group specialised register, the Cochrane Controlled trials register, EMBASE, MEDLINE, CINAHL and PSYCHLIT from 1966 to January 2002, without language restriction. Reference lists of articles were scanned and we contacted experts in the area to find other relevant studies. SELECTION CRITERIA: Randomised or controlled trials that compared sensory stimulation programmes with standard rehabilitation in patients in coma or vegetative state. DATA COLLECTION AND ANALYSIS: Abstracts and papers found were screened by one reviewer. Three reviewers independently identified relevant studies, extracted data and assessed study quality resolving disagreement by consensus. MAIN RESULTS: Three studies were identified with 68 patients in total. The overall methodological quality was poor and studies differed widely in terms of outcomes measures, study design and conduct. We therefore did not carry out any quantitative synthesis but reviewed results of available studies qualitatively. REVIEWER'S CONCLUSIONS: This systematic review indicates that there is no reliable evidence to support, or rule out, the effectiveness of multisensory programmes in patients in coma or vegetative state. PMID- 12076411 TI - Riluzole for amyotrophic lateral sclerosis (ALS)/motor neuron disease (MND). AB - BACKGROUND: Riluzole has been approved for treatment of patients with amyotrophic lateral sclerosis in many countries but not all. Questions persist about its clinical utility because of high cost, modest efficacy and concern over adverse effects. OBJECTIVES: To examine the efficacy of riluzole in prolonging survival, and in delaying the use of surrogates (tracheostomy and mechanical ventilation) to sustain survival. SEARCH STRATEGY: Search of the Cochrane Neuromuscular Disease Group Register for randomized trials and enquiry from authors of trials, Aventis (manufacturer of riluzole) and other experts in the field. The most recent search was May, 2001 SELECTION CRITERIA: Types of studies: randomized trials TYPES OF PARTICIPANTS: adults with a diagnosis of amyotrophic lateral sclerosis Types of interventions: treatment with riluzole or placebo Types of outcome measures: Primary: pooled hazard ratio of tracheostomy-free survival over all time points with riluzole 100 mg. Secondary: per cent mortality as a function of time with riluzole 100 mg and other doses of riluzole; neurologic function, quality of life, muscle strength and adverse events. DATA COLLECTION AND ANALYSIS: We identified four eligible randomized trials. Each reviewer graded them for methodological quality. Data extraction was performed by a single reviewer and checked by two others. We obtained some missing data from investigators and regulatory agencies. We performed meta-analyses with Review Manager 4.1 software using a fixed effects model. A test of drug efficacy was based on the Parmar pooled hazard ratio. MAIN RESULTS: The three trials examining tracheostomy-free survival included a total of 876 riluzole treated patients and 406 placebo treated patients. The data for tracheostomy-free survival was not available from the fourth trial. The methodological quality was acceptable and the three trials were easily comparable, although one trial included older patients in more advanced stages of amyotrophic lateral sclerosis. Riluzole 100 mg per day provided a benefit for the homogeneous group of patients in the first two trials (p=0.039, hazard ratio 0.80, 95% confidence interval 0.64 to 0.99) and there was no evidence of heterogeneity (p=0.33). When the third trial (which included older and more seriously affected patients) is added, there is evidence of heterogeneity (p<0.0001) and the random effects model, which takes this into account results in the overall treatment effect estimate falling just short of significance (p=0.056, hazard ratio 0.84, 95% confidence interval 0.70 to 1.01). This represents a 9% gain in the probability of surviving one year (57% in the placebo and 66% in the riluzole group). In secondary analyses of survival at separate time points, there was a significant survival advantage with riluzole 100 mg at six, nine, 12 and 15 months, but not at three or 18 months. There was a small beneficial effect on both bulbar and limb function, but not on muscle strength. There were no data on quality of life, but patients treated with riluzole remained in a more moderately affected health state significantly longer than placebo-treated patients (weighted mean difference 35.5 days, 95% confidence interval 5.9 to 65.0). A threefold increase in serum alanine transferase was more frequent in riluzole treated patients than controls (weighted mean difference 2.69, 95% confidence interval 1.65 to 4.38). REVIEWER'S CONCLUSIONS: Riluzole 100 mg daily is reasonably safe and probably prolongs survival by about two months in patients with amyotrophic lateral sclerosis. More studies are needed, especially to clarify its effect in older patients (over 75 years), and those with more advanced disease. PMID- 12076412 TI - Pneumococcal vaccines for preventing otitis media. AB - BACKGROUND: Acute otitis media (AOM) is one of the most common diseases in early infancy and childhood. Long term effects of recurrent episodes of otitis media, rapid emergence of drug resistant bacteria associated with AOM worldwide and huge estimated direct and indirect annual costs associated with otitis media have emphasized the need for an effective vaccination program to prevent episodes of AOM. OBJECTIVES: The object of this review was to assess the effect of pneumococcal vaccination in preventing AOM in children up to 12 years of age. SEARCH STRATEGY: We searched the Cochrane Acute Respiratory Infection Group specialised register (last update, 26th April 2001), the Cochrane Library (Issue 4, 2000), MEDLINE (January 1966-August 2000) and reference list of all studies and review articles retrieved. We also contacted two vaccine manufacturers and first or corresponding authors of some included studies. SELECTION CRITERIA: Randomised controlled clinical trials of pneumococcal vaccination with prevention of AOM as outcome in children aged 12 years or younger and a follow-up of at least six months. DATA COLLECTION AND ANALYSIS: Five reviewers independently assessed trial quality and two reviewers extracted data. Two study authors were contacted. MAIN RESULTS: Eight trials on pneumococcal polysaccharide vaccine (PPV) and two trials on pneumococcal conjugate vaccine (PCV) were included. The highest efficacy of PPV was found in children aged 24 months and older: the rate ratio after adjustment for study was 0.833 [95%CI: 0.625-0.970]. The PPV has little effect on the prevention of AOM in children without documented prior episodes of AOM and only a moderate effect in the group of children with documented AOM episodes prior to vaccination. The results of the two PCV trials in healthy infants, which followed children from the age of two months until two years of age, could not be pooled because of lack of data. Both studies showed that the risk of recurrent disease decreased with 9% in the group of children receiving the PCV together with other childhood vaccinations at 2,4,6 and 14 months of age: Study Black et al 2000 : risk ratio=0.91[95%CI:0.86-0.96]; Study Eskola et al 2001: risk ratio=0.90 [95%CI:0.73-1.12]. REVIEWER'S CONCLUSIONS: Based on the currently available results of the effectiveness of pneumococcal vaccination for the prevention of AOM, a large scale use of pneumococcal vaccination for this indication is not recommended. The results of currently ongoing trials could provide more information whether pneumococcal vaccines are effective in specific high-risk (otitis-prone) populations. PMID- 12076413 TI - Endometrial destruction techniques for heavy menstrual bleeding. AB - BACKGROUND: Heavy menstrual bleeding (HMB) is a significant health problem in premenopausal women that can reduce quality of life and cause anaemia. First line therapy has traditionally been medical therapy but this is not always completely effective. Hysterectomy, often used after the failure of medical therapy, is 100% effective but is risky, costly and can cause complications. Endometrial ablation is less invasive, less costly and preserves the uterus. A large number of techniques have been developed to "ablate" (remove) the lining of the endometrium. The gold standard techniques (laser, transcervical resection of the endometrium and rollerball) require visualisation of the uterus with a hysteroscope and, although safe, require skilled surgeons. A number of newer techniques have recently been developed, most of which can be performed blind and are less time consuming. Many of these techniques are still under development, refinement and investigation. OBJECTIVES: To compare the efficacy, safety and acceptability of methods used to destroy the endometrium to reduce HMB in premenopausal women. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (issue 4, 2001), Medline (1966 to September 2001), EmBase (1980 to August 2001), Current Contents (1993 to week 38, 2001), Biological Abstracts (1980 to June 2001), Psyclit (1967 to August 2001) and Cinahl (1982 to July 2001). We also searched the specialised register of the Cochrane Menstrual Disorders and Subfertility Group (August 2001). We also searched reference lists of articles and contacted pharmaceutical companies and experts in the field. SELECTION CRITERIA: Randomised controlled trials comparing endometrial ablation techniques in women with a complaint of heavy menstrual bleeding without uterine pathology. The outcomes included reduction of heavy menstrual bleeding, improvement in quality of life, operative outcomes, satisfaction with outcome, complications and need for further surgery. DATA COLLECTION AND ANALYSIS: The two reviewers independently selected trials for inclusion, assessed trials for quality and extracted data. Attempts were made to contact authors for clarification of data in some trials. Adverse events were only assessed if they were separately measured in the included trials. MAIN RESULTS: In comparing hysteroscopic techniques, the vaporising electrode procedure was less difficult to perform (OR=0.25, 95%CI 0.1, 0.7) and had less fluid deficit (WMD=-258mls, 95% CI -342.1, -174.0) than TCRE. The odds of fluid overload and equipment failure were higher ((OR=5.2, 95% CI 1.5, 18.4) and (OR=6.0, 95% CI 1.7, 20.9) respectively) for those women having laser as compared to TCRE (transcervical resection of the endometriuim). In comparing traditional hysteroscopic endometrial ablation with the newer 2nd generation techniques overall, the newer techniques took less time to perform (WMD=-11mins, 95% CI -18.6, -2.6) and were more likely to be performed under local anaesthesia (OR=7.6, 95% CI 1.1, 52.7) but had a greater chance of equipment failure (OR=4.1, 95% CI 1.1, 15.0). The reduction in heavy bleeding did not differ significantly between any of the groups. REVIEWER'S CONCLUSIONS: Endometrial ablation techniques continue to play an important role in the management of HMB. The rapid development of a number of new methods of endometrial destruction has made systematic comparisons between methods and with the "gold standard" of TCRE difficult. Most of the newer techniques are performed blind and are technically easier than hysteroscopy-based methods. Overall, the existing evidence suggests success rates and complication profile of newer techniques of ablation compares favourably with TCRE, although technical difficulties with new equipment need to be ironed out. PMID- 12076414 TI - Melatonin for the prevention and treatment of jet lag. AB - BACKGROUND: : Jet-lag commonly affects air travellers who cross several time zones. It results from the body's internal rhythms being out of step with the day night cycle at the destination. Melatonin is a pineal hormone that plays a central part in regulating bodily rhythms and has been used as a drug to re-align them with the outside world. OBJECTIVES: : To assess the effectiveness of oral melatonin taken in different dosage regimens for alleviating jet-lag after air travel across several time zones. SEARCH STRATEGY: : We searched the Cochrane Controlled Trials Register, MEDLINE, EMBASE, PsychLit and Science Citation Index electronically, and the journals 'Aviation, Space and Environmental Medicine' and 'Sleep' by hand. We searched citation lists of relevant studies for other relevant trials. We asked principal authors of relevant studies to tell us about unpublished trials. Reports of adverse events linked to melatonin use outside randomised trials were searched for systematically in 'Side Effects of Drugs' (SED) and SED Annuals, 'Reactions Weekly', MEDLINE, and the adverse drug reactions databases of the WHO Uppsala Monitoring Centre (UMC) and the US Food & Drug Administration. SELECTION CRITERIA: : Randomised trials in airline passengers, airline staff or military personnel given oral melatonin, compared with placebo or other medication. Outcome measures should consist of subjective rating of jet-lag or related components, such as subjective wellbeing, daytime tiredness, onset and quality of sleep, psychological functioning, duration of return to normal, or indicators of circadian rhythms. DATA COLLECTION AND ANALYSIS: : Ten trials met the inclusion criteria. All compared melatonin with placebo; one in addition compared it with a hypnotic, zolpidem. Nine of the trials were of adequate quality to contribute to the assessment, one had a design fault and could not be used in the assessment. Reports of adverse events outside trials were found through MEDLINE, 'Reactions Weekly', and in the WHO UMC database. MAIN RESULTS: : Nine of the ten trials found that melatonin, taken close to the target bedtime at the destination (10pm to midnight), decreased jet lag from flights crossing five or more time zones. Daily doses of melatonin between 0.5 and 5mg are similarly effective, except that people fall asleep faster and sleep better after 5mg than 0.5mg. Doses above 5mg appear to be no more effective. The relative ineffectiveness of 2mg slow-release melatonin suggests that a short-lived higher peak concentration of melatonin works better. Based on the review, the number needed to treat (NNT) is 2. The benefit is likely to be greater the more time zones are crossed, and less for westward flights. The timing of the melatonin dose is important: if it is taken at the wrong time, early in the day, it is liable to cause sleepiness and delay adaptation to local time. The incidence of other side effects is low. Case reports suggest that people with epilepsy, and patients taking warfarin may come to harm from melatonin. REVIEWER'S CONCLUSIONS: : Melatonin is remarkably effective in preventing or reducing jet-lag, and occasional short-term use appears to be safe. It should be recommended to adult travellers flying across five or more time zones, particularly in an easterly direction, and especially if they have experienced jet-lag on previous journeys. Travellers crossing 2-4 time zones can also use it if need be. The pharmacology and toxicology of melatonin needs systematic study, and routine pharmaceutical quality control of melatonin products must be established. The effects of melatonin in people with epilepsy, and a possible interaction with warfarin, need investigation. PMID- 12076415 TI - Safety education of pedestrians for injury prevention. AB - BACKGROUND: Each year about one million people die and about 10 million are seriously injured on the world's roads. Educational measures to teach pedestrians how to cope with the traffic environment are considered to be an essential component of any prevention strategy, and pedestrian education has been recommended in many countries. However, as resources available for road safety are limited, a key question concerns the relative effectiveness of different prevention strategies. OBJECTIVES: To quantify the effectiveness of pedestrian safety education programmes in preventing pedestrian-motor vehicle collisions. SEARCH STRATEGY: We searched the Injuries Group specialised register, Cochrane Controlled Trials Register, TRANSPORT, MEDLINE, EMBASE, ERIC, PSYCHLIT, SPECTR, and the WHO database on the Internet. We checked reference lists of relevant reviews and papers and contacted experts in the field. Most database searching was conducted in 1999. SELECTION CRITERIA: Randomised controlled trials of safety education programmes for pedestrians of all ages. DATA COLLECTION AND ANALYSIS: One reviewer screened records. Two reviewers independently extracted data and assessed methodological quality of trials. Because of differences in the types of interventions and outcome measures used in the trials, meta-analyses were not carried out. MAIN RESULTS: We found 15 randomised-controlled trials of pedestrian safety education programmes, conducted between 1976 and 1997. Methodological quality of the included trials was generally poor. Allocation concealment was adequate in three trials, outcome assessment was blinded in eight, and in most of the studies large numbers of participants were lost to follow-up. Study participants were children in 14 studies and institutionalised adults in one. Eight studies involved the direct education of participants, seven used parents as educators. No trials were conducted in a developing country and there were none of pedestrian safety training in the elderly. None of the included trials assessed the effect of pedestrian safety education on the occurrence of pedestrian injury but six trials assessed the effect on observed behaviour. Some of these trials showed evidence of behavioural change following pedestrian safety education but it is difficult to predict what effect this might have on pedestrian injury risk. REVIEWER'S CONCLUSIONS: Pedestrian safety education can result in improvement in children's knowledge and can change observed road crossing behaviour but whether this reduces the risk of pedestrian motor vehicle collision and injury occurrence is unknown. There is evidence that changes in safety knowledge and observed behaviour decline with time suggesting that safety education must be repeated at regular intervals. PMID- 12076416 TI - Surgical versus non-surgical treatment for carpal tunnel syndrome. AB - BACKGROUND: Carpal tunnel syndrome is the clinical condition resulting from the entrapment of the median nerve in the wrist. It has been accepted as the most frequent entrapment neuropathy. The most common symptoms are tingling, numbness, and pain in the hand that may radiate to the forearm or shoulder. There may be weakness and atrophy of the thenar muscles associated with sensory loss in the affected fingers. There is no universally accepted therapy for carpal tunnel syndrome. Surgical treatment is widely preferred to non-surgical or conservative therapies for overtly symptomatic patients, while mild cases are usually not treated. OBJECTIVES: The objective of this review is to compare the efficacy of surgical treatment of carpal tunnel syndrome with non-surgical treatment in improving clinical outcome. SEARCH STRATEGY: We searched the Cochrane Neuromuscular Disease Group register for randomised or quasi-randomised trials as well as MEDLINE, EMBASE and LILACS (to July 2001). We checked the bibliographies in relevant papers and contacted the authors to obtain information about other published or unpublished studies. SELECTION CRITERIA: All randomised and quasi randomised controlled trials comparing any surgical and any non-surgical therapies. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed eligibility of the trials. MAIN RESULTS: Only one randomised controlled trial was found. It included 22 female patients, 11 allocated to surgical section of the anterior carpal ligament and 11 to splinting for one month. The trial was not blinded and it is not clear if the allocation was properly concealed. Data reported allowed an intention-to-treat analysis on two secondary outcomes. The results favour surgery for both of them. There was a significant clinical improvement at one year follow-up in 10 out of 11 patients allocated to surgery and two out of 11 allocated to splinting (relative risk 5.00, 95% confidence interval 1.41, 17.76). Eight out of 11 patients allocated to splinting required surgery during follow-up, compared with apparently no re operation in the surgical group (relative risk 0.06, 95% confidence interval 0.00, 0.91). REVIEWER'S CONCLUSIONS: Surgical treatment of carpal tunnel syndrome seems to be better than splinting. There is a need for randomised controlled trials comparing surgical and non-surgical therapies for carpal tunnel syndrome, particularly in patients with mild symptoms in whom there is greater uncertainty concerning the balance of risks versus benefit of surgical therapy. PMID- 12076417 TI - Prophylaxis for venous thromboembolic disease in pregnancy and the early postnatal period. AB - BACKGROUND: Venous thromboembolic disease (TED), although very rare, is a major cause of maternal mortality and morbidity, hence methods of prophylaxis are often used for women at risk. This may include women delivered by caesarean section, those with a personal or family history of TED and women with inherited or acquired thrombophilias (conditions that predispose people to thrombosis). Many methods of prophylaxis carry a risk of side effects, and as the risk of thromboembolic disease is low, it is possible that the benefits of thromboprophylaxis may be outweighed by harm. Current guidelines for clinical practice are based on expert opinion only, rather than high quality evidence from randomised trials. OBJECTIVES: To determine the effects of thromboprophylaxis in association with pregnancy in women who are pregnant or have recently delivered on the incidence of venous thromboembolic disease and side effects. SEARCH STRATEGY: The register of trials maintained by the Cochrane Pregnancy and Childbirth Group, the Cochrane Controlled Trials Register, MEDLINE, EMBASE and bibliographies of reviews. Date of last search: January 2002. SELECTION CRITERIA: Randomised trials comparing one method of thromboprophylaxis with placebo or no treatment, and randomised trials comparing two (or more) methods of thromboprophylaxis. DATA COLLECTION AND ANALYSIS: Data were extracted independently by all reviewers. Discrepancies were resolved by discussion. MAIN RESULTS: Eight trials involving 649 women were included. Four of them compared methods of antenatal prophylaxis; low molecular weight versus unfractionated heparin (two studies), aspirin plus heparin versus aspirin alone (one study), and unfractionated heparin versus no treatment (one study). Four studies assessed postnatal prophylaxis after caesarean section; one compared hydroxyethyl starch with unfractionated heparin, two compared heparin with placebo (one low molecular weight heparin, one unfractionated heparin) and the other compared unfractionated heparin with low molecular weight heparin. It was not possible to assess the effects of any of these interventions on most outcomes, especially rare outcomes such as death, thromboembolic disease and osteoporosis, because of small sample sizes and the small number of trials making the same comparisons. REVIEWER'S CONCLUSIONS: There is insufficient evidence on which to base recommendations for thromboprophylaxis during pregnancy and the early postnatal period. Large scale randomised trials of currently-used interventions should be conducted. PMID- 12076418 TI - Mobilisation strategies after hip fracture surgery in adults. AB - BACKGROUND: Post-operative care programmes after hip fracture surgery include strategies for mobilisation, such as early weight bearing, gait retraining and other physical therapy interventions. OBJECTIVES: To evaluate the effects of different mobilisation strategies and programmes after hip fracture surgery. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register (to February 2002), MEDLINE (1966 to February 2002), the National Research Register (Issue 2, 2001), Current Controlled Trials, conference proceedings and reference lists of articles. SELECTION CRITERIA: All randomised or quasi-randomised trials comparing different mobilisation strategies/programmes after hip fracture surgery. DATA COLLECTION AND ANALYSIS: The reviewers independently assessed trial quality, using a ten item scale, and extracted data. Wherever appropriate and possible, the data are presented graphically. MAIN RESULTS: Our first update included one new study. Again, each of the now five included trials evaluated a different intervention. Most trials had poor and / or poorly reported trial methodology. One trial of 100 patients which compared twice daily with once daily physiotherapy reported a lack of demonstrable difference in recovery of the two patient groups at nine weeks follow-up. Only limited outcome data were available for checking this. One trial of 80 patients evaluated a quadriceps muscle strengthening exercise programme. Improved mobility, leg extension power and Barthel score were reported for the intervention group. A treadmill gait retraining programme was compared with a conventional gait retraining programme in 40 patients. More patients in the treadmill group had recovered their pre-fracture level of mobility by the time of hospital discharge, which tended to happen earlier than for the control group. Neither of these differences were statistically significant. One trial of 24 patients compared neuromuscular stimulation of the quadriceps muscle with placebo stimulation. No data were available to test the claims that neuromuscular stimulation improved the recovery of mobility, assessed up to 13 weeks. One trial involving 273 patients with a displaced intracapsular fracture treated by internal fixation compared weight bearing at two weeks after surgery with delayed weight bearing at 12 weeks after surgery. From the limited data available, there were no statistically significant differences between the two methods of treatment for the outcomes of non-union, mortality and overall unfavourable outcome at one year. REVIEWER'S CONCLUSIONS: There is insufficient evidence from randomised trials to determine the effects of more frequent physiotherapy, quadriceps strengthening exercises, treadmill gait retraining, or neuromuscular stimulation after hip fracture surgery. There is also insufficient evidence to determine the effects of early weight bearing after the internal fixation of an intracapsular proximal femoral fracture. PMID- 12076419 TI - Treatment of infantile spasms. AB - BACKGROUND: Infantile spasms (West's Syndrome) is a syndrome which includes a peculiar type of epileptic seizure, usually hypsarrhythmia and in the majority of patients, psychomotor retardation. It remains poorly understood and despite modern imaging techniques an underlying cause is often not found. Little is known about their pathophysiological basis and treatment remains problematic. OBJECTIVES: To compare the effects of single drugs used to treat infantile spasms upon long-term psychomotor development, subsequent epilepsy, control of the spasms and side effects. SEARCH STRATEGY: Our search included the Cochrane Epilepsy Group trials register, MEDLINE (1960 to 2001), EMBASE (1981 to 2001), contacting pharmaceutical companies and appeals at international conferences. SELECTION CRITERIA: Randomized controlled trials (RCTs) of the administration of drugs to people with infantile spasms. DATA COLLECTION AND ANALYSIS: Three reviewers independently selected trials for inclusion and extracted data. Study quality and potential sources of heterogeneity were assessed. Outcomes included cessation of spasms, time to cessation of spasms, participants with cessation of spasms remaining spasm free, reduction in spasms, resolution of hypsarrhythmia, subsequent epilepsy rates and side effects. MAIN RESULTS: Ten small RCTs were included. In total these studies recruited just 335 participants and tested eight different drugs. Overall, studies were of poor methodological quality. No study assessed long-term psychomotor development or the development of other seizure types. One small study found vigabatrin to be more efficacious than hydrocortisone in stopping infantile spasms in a group of people with tuberous sclerosis. One underpowered study showed a trend for vigabatrin to be more efficacious than placebo in stopping infantile spasms, two small studies when combined showed ACTH to be more efficacious than low-dose prednisone (2mg/kg). It was not possible to compare reduction in the number of spasms between the different treatments because of differences in methods of analysis. Overall, only nine participants were reported to have been withdrawn from the trial treatments due to side effects and two deaths were reported. REVIEWER'S CONCLUSIONS: We found no reliable evidence that any of the treatments assessed were more efficacious than any other. Few studies considered psychomotor development or subsequent seizure rates as outcomes and none had long term follow-up. Further trials with larger numbers of participants, and longer follow-up are required. PMID- 12076420 TI - Posture and fluids for preventing post-dural puncture headache. AB - BACKGROUND: Potentially disabling postural headache occurs after dural puncture in 1-70% of patients. It has been suggested that such headaches may be less common if patients routinely have a period of bed rest or receive supplementary fluids after the procedure. OBJECTIVES: To assess the effects on post dural puncture headache of a period of bed rest versus early mobilisation, of different positions during a period of bed rest, and of administering supplementary fluids after the procedure. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (Cochrane Library, Issue 4, 2000), MEDLINE (January 1994-December 1998), and EMBASE (January 1980-December 1998). We also searched the reference lists of articles identified electronically, and contacted trial authors for information about other potentially relevant studies. Date of the most recent search: December 2000. SELECTION CRITERIA: We sought randomised, unconfounded trials, among all types of patients, that compared the effects on post-dural puncture headache of: bed rest versus early mobilisation; head-down tilt versus horizontal or prone versus supine positions during bed rest; or the administration of supplementary fluids versus control. DATA COLLECTION AND ANALYSIS: One reviewer extracted data from the reports of all trials considered eligible for inclusion. The authors of included studies were invited to check the information extracted and provide any details that were unavailable in the published reports. Intention to-treat analyses were performed. MAIN RESULTS: Eleven trials among 1723 patients compared either bed rest with immediate mobilisation or a longer versus a shorter period of bed rest. There was a non-significant relative increase in the odds of the primary outcome of postural headache among patients allocated more rather than less bed rest (196/639 [31%] bed rest versus 169/615 [27%] early mobilisation; odds ratio [OR] 1.21; 95% confidence interval [CI] 0.94 to 1.55). Analyses confined to the methodologically most rigorous trials gave similar results. Only two trials among 126 patients compared different positions during bed rest. No statistically significant differences were found, but small numbers made the comparisons imprecise. One trial among 100 patients assessed fluid supplementation. Again, the numbers of patients and outcome events were small, and indicated the possibility of both reduced and increased postural headache with additional fluids. REVIEWER'S CONCLUSIONS: There is no good evidence from randomised trials to suggest that routine bed rest after dural puncture is beneficial. The role of fluid supplementation in the prevention of post-dural puncture headache remains uncertain. PMID- 12076421 TI - Epidural blood patching for preventing and treating post-dural puncture headache. AB - BACKGROUND: Dural puncture is a common procedure, but leakage of CSF from the resulting dural defect may cause postural headache after the procedure, and this can be disabling. Injecting an epidural blood patch around the site of the defect may stop this leakage, and so may have a role in preventing or treating post dural puncture headache. OBJECTIVES: To assess the possible benefits and harms of epidural blood patching in both the prevention and the treatment of post-dural puncture headache. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (Cochrane Library, Issue 4, 2000), MEDLINE (January 1994 to December 1998), and EMBASE (January 1980 to December 1998). We also searched the reference lists of relevant articles identified electronically, and asked both the authors of all included trials and colleagues with an interest in this area to let us know of any other potentially relevant studies not already identified. Date of last search: December 2000. SELECTION CRITERIA: We sought all properly randomised, unconfounded trials that compared epidural blood patch versus no epidural blood patch in the prevention or treatment of post-dural puncture headache among all types of patients undergoing dural puncture for any reason. The primary outcome of effectiveness was postural headache. DATA COLLECTION AND ANALYSIS: One reviewer extracted details of trial methodology and outcome data from the reports of all trials considered eligible for inclusion. We invited the authors of all such trials both to check the information extracted and to provide any details that were unavailable in the published reports. Intention-to-treat analyses were performed using the Peto O-E method. Information about adverse effects (post-dural puncture backache, epidural infection and lower limb paraesthesia) was also extracted. MAIN RESULTS: Three trials (77 patients) were eligible for inclusion. Methodological details were generally incomplete. Although the results of our analyses suggested that both prophylactic and therapeutic epidural blood patching may be of benefit, the very small numbers of patients and outcome events, as well as uncertainties about trial methodology, precluded reliable assessments of the potential benefits and harms of this intervention. REVIEWER'S CONCLUSIONS: Further, adequately powered, randomised trials (including at least a few hundred patients) are required before reliable conclusions can be drawn about the role of epidural blood patching in the prevention and treatment of post-dural puncture headache. PMID- 12076422 TI - Aerobic exercise interventions for adults living with HIV/AIDS. AB - BACKGROUND: The profile of HIV infection is constantly changing. Although once viewed as an illness progressing to death, among those with access to antiretroviral therapy, HIV infection can now present as a disease with an uncertain natural history, perhaps a chronic manageable disease for some. Thus, the needs of persons infected with HIV have increasingly included management of impairments, disabilities and handicaps. Exercise is a key management strategy employed by rehabilitation professionals to address these issues and assist persons living with HIV/AIDS. Exercise has been shown to improve strength, cardiovascular function, and psychological status in seronegative populations (~~Bouchard 1993~~) but what are the effects of exercise for adults living with HIV infection? If the risks and benefits of exercise for people living with HIV infection are better understood, appropriate exercise prescription may be practiced by health care providers, and may enhance the effectiveness of HIV management, thus improving the overall outcome for adults living with HIV infection. OBJECTIVES: To examine the effect of aerobic exercise interventions on cardiopulmonary, immunological/virological and psychological parameters in adults living with HIV infection. SEARCH STRATEGY: To identify the appropriate studies, we conducted a search using MEDLINE, EMBASE, SCIENCE CITATION INDEX, AIDSLINE, CINAHL, HEALTHSTAR, PSYCHLIT, SOCIOFILE, SCI, SSCI, ERIC and DAI. We also reviewed abstracts from international AIDS, ICAAC, and other major meetings. Reference lists from pertinent articles and books and personal contact with authors were also used, as were Collaborative Review Group databases and results of hand searching of targeted journals. All languages were included. Searches for the original review covered the period from 1980 to July 1999. For the purposes of this update, an additional literature search, following the same identification of studies criteria as listed above, was conducted from August 1999 to January 2001. SELECTION CRITERIA: To be selected, studies had to be randomized controlled trials involving HIV+ adults 18 years of age or older and had to include at least one group randomized to receive aerobic exercise performed at least three times/week for at least four weeks. DATA COLLECTION AND ANALYSIS: Data on study design, participants, interventions, and outcomes were extracted from the reports onto specifically designed data collection forms by at least two reviewers. MAIN RESULTS: A total of eight studies (six from the original search and two from the updated search) were identified that satisfied the eligibility criteria. The main results indicated that performing constant or interval aerobic exercise, or a combination of constant aerobic exercise and progressive resistive exercise for at least 20 minutes, at least three times per week for four weeks appears to be safe and may lead to clinically significant improvements in cardiopulmonary fitness. Furthermore, individual studies suggest that aerobic exercise may improve psychological well-being for adults living with HIV/AIDS. These findings are limited to those who continued to exercise and for whom there was adequate follow-up data. REVIEWER'S CONCLUSIONS: Aerobic exercise appears to be safe and may be beneficial for adults living with HIV/AIDS. These findings are limited by the small sample sizes and large drop-out rates of the included studies. Future research would benefit from an increased attention to participant follow-up and intention-to-treat analysis. Further research is required to determine the optimal parameters of aerobic exercise and stage of disease in which aerobic exercise may be most beneficial for adults living with HIV infection. PMID- 12076423 TI - Intravenous immunoglobulin for chronic inflammatory demyelinating polyradiculoneuropathy. AB - BACKGROUND: Chronic inflammatory demyelinating polyradiculoneuropathy is an immune mediated disorder characterised by progressive or relapsing symmetrical motor or sensory symptoms and signs in more than one limb, developing over at least two months. It may cause prolonged periods of disability and even death. Several uncontrolled studies have suggested a beneficial effect of intravenous immunoglobulin. OBJECTIVES: To review systematically the evidence from randomised controlled trials concerning the efficacy and safety of intravenous immunoglobulin in chronic inflammatory demyelinating polyradiculoneuropathy. SEARCH STRATEGY: We used the Search Strategy of the Cochrane Neuromuscular Disease Review Group to search the Disease Group register and other databases for randomised controlled trials from 1985 onwards. SELECTION CRITERIA: Randomised controlled studies examining the effects of any dose of intravenous immunoglobulin versus placebo, plasma exchange or corticosteroids in patients with definite or probable chronic inflammatory demyelinating polyradiculoneuropathy. Outcome measures had to include one of the following: a disability score, the Medical Research Council sum score, electrophysiological data or walking distance. Studies which reported the frequency of adverse effects were used to assess the safety of treatment. DATA COLLECTION AND ANALYSIS: Two reviewers independently reviewed literature searches to identify potentially relevant trials, scored their quality and extracted data independently. For dichotomous data, we calculated relative risks, and for continuous data, effect sizes (for definition see statistical analysis section) and weighted pooled effect sizes. Statistical uncertainty was expressed in 95% confidence intervals. Sensitivity analysis excluding studies with quality scores below A 0.50 and below B 0.75 was planned but not performed as all studies had quality scores above 0.75. MAIN RESULTS: Six randomised controlled trials were considered eligible including 170 patients. Four studies on 113 patients compared intravenous immunoglobulin against placebo. One trial with 17 patients compared intravenous immunoglobulin with plasma exchange in a cross-over design and one trial compared intravenous immunoglobulin with prednisolone in 32 patients. A significantly higher proportion of patients improved in disability within one month after the onset of intravenous immunoglobulin treatment as compared with placebo (relative risk 3.17, 95% confidence interval 1.74 to 5.75). Whether all these improvements are equally clinically relevant cannot be deduced from this analysis because each trial used a different disability scale with a unique definition of a significant improvement. To overcome this problem an attempt was made to transform the various disability scales to the modified Rankin score. In three trials including 87 patients this transformation could be carried out. A significantly higher proportion of patients improved one point after intravenous immunoglobulin treatment compared to placebo (relative risk 2.47, 95% confidence interval 1.02 to 6.01). The effect size for change of mean disability score at six weeks comparing intravenous immunoglobulin with plasma exchange revealed no difference between the two therapies (effect size -0.07, 95% confidence interval -0.76 to 0.63.) The proportion of patients with a significant improvement did not differ significantly between prednisolone and intravenous immunoglobulin (relative risk of 0.91 (95% CI 0.50 to 1.68). Also, no difference in mean improvement on the disability scale was found at two weeks (effect size -0.12, 95% confidence interval -0.68 to 0.45) or six weeks (effect size -0.07, 95% confidence interval 0.63 to 0.50) between prednisolone and intravenous immunoglobulin. There were no statistically significant differences in frequencies of side effects between the three types of treatment. REVIEWER'S CONCLUSIONS: The evidence from randomised controlled trials shows that intravenous immunoglobulin improves disability for at least two to six weeks compared with placebo, with a number needed to treat of three. During this period it has similar efficacy to plasma exchange and oral prednisolone. Since intravenous immunoglobulin, plasma exchange and prednisolone seem to be equally effective, it is currently uncertain which of these treatments should be the first choice. Cost, side effects, duration of treatment, dependency on regular hospital visits and ease of administration all have to be considered before such a decision can be made. PMID- 12076424 TI - Plasma exchange for Guillain-Barre syndrome. AB - BACKGROUND: Guillain-Barre syndrome is an acute symmetric, usually ascending and usually paralysing illness, due to inflammation of peripheral nerves. It is thought to be caused by autoimmune factors, such as antibodies. Plasma exchange removes antibodies and other potentially injurious factors from the blood stream. It involves connecting the patient's blood circulation to a machine which exchanges the plasma for a substitute solution, usually albumin. Several studies have evaluated plasma exchange for Guillain-Barre syndrome. OBJECTIVES: To systematically review the evidence concerning the efficacy of plasma exchange for treating Guillain-Barre syndrome. SEARCH STRATEGY: Search of the Cochrane Neuromuscular Disease Trial Register for randomised trials concerning plasma exchange in Guillain-Barre syndrome, search of the bibliographies of identified papers and enquiry from the authors of the papers. SELECTION CRITERIA: Randomised and quasi-randomised trials of plasma exchange versus sham exchange or supportive treatment. DATA COLLECTION AND ANALYSIS: Potentially relevant papers were scrutinised by two reviewers and the selection of eligible studies was agreed by them and a third reviewer. Data were extracted by one reviewer and checked by a second reviewer. Some missing data were obtained from the authors of studies. MAIN RESULTS: Six eligible trials concerning 649 patients were identified, all comparing plasma exchange versus supportive treatment alone. Primary outcome measures ~bullet~Time to recover walking with aid In the only two trials for which this measure was reported, the median time to recover this ability was faster in the plasma exchange than the control group. ~bullet~Time to onset of motor recovery in mildly affected patients In the one trial for which this measure was available, the time was significantly shortened in the plasma exchange group. Secondary outcome measures ~bullet~Improvement in disability grade at four weeks In five trials, there were significantly more patients who had improved by one disability grade or more in the plasma exchange group as compared to the control group. Patients treated with plasma exchange fared significantly better in the following secondary outcome measures: time to recover walking without aid, percentage of patients requiring artificial ventilation, duration of ventilation, full muscle strength recovery after one year, and severe sequelae after one year. There were less patients with infectious events and cardiac arrhythmias in the plasma exchange than the control group. Subgroup analyses Plasma exchange was beneficial in patients with mild, moderate and severe (needing ventilation) Guillain-Barre syndrome. It was beneficial in patients with a disease duration of seven or less days and also in those with disease lasting more than seven days. However, in the only trial that enrolled patients up to 30 days from disease onset, the benefit of plasma exchange in patients treated after seven days was less apparent. Type of treatment Single studies showed that two plasma exchanges were significantly superior to none for mild Guillain-Barre syndrome and four to two for moderate Guillain-Barre syndrome, but that six were not superior to four for severe Guillain-Barre syndrome requiring ventilation. One study suggested that continuous flow plasma exchange was significantly superior to intermittent flow. Another study found no significant difference between the two techniques. The same study found a significantly higher rate of adverse events with fresh frozen plasma as the replacement fluid than albumin. Plasma exchange compared with cerebrospinal fluid filtration A single trial comparing these two treatments did not show any difference in outcomes but was too small to demonstrate equivalence. REVIEWER'S CONCLUSIONS: Plasma exchange is the first and only treatment that has been proven to be superior to supportive treatment alone in Guillain-Barre syndrome. Consequently, plasma exchange should be regarded as the treatment against which new treatments, such as intravenous immunoglobulin, should be judged. In mild Guillain-Barre syndrome two sessions of plasma exchange are superior to none. In moderate Guillain-Barre syndrome four sessions are superior to two. In severe Guillain-Barre syndrome six sessions are no better than four. Continuous flow plasma exchange machines may be superior to intermittent flow machines and albumin to fresh frozen plasma as the exchange fluid. Plasma exchange is more beneficial when started within seven days after disease onset rather than later, but was still beneficial in patients treated up to 30 days after disease onset. The value of plasma exchange in children less than 12 years old is not known. There is insufficient evidence to determine whether cerebrospinal fluid filtration is equivalent to plasma exchange. PMID- 12076425 TI - Opioid antagonists for alcohol dependence. AB - BACKGROUND: The results from animal studies suggest that opioid antagonists may prevent the reinforcing effects of alcohol consumption. Based on the results of those animal studies, some opioid antagonists, such as, naltrexone, nalmefene, have been studied for their benefits in treating alcohol dependence. OBJECTIVES: To determine the effectiveness of opioid antagonists in attenuating or preventing the recommencement of alcohol consumption in patients with alcohol dependence in comparison to placebo, other medications and psychosocial treatments. In addition, discontinuation rate, death, patient satisfaction, functioning, health related quality of life and economic outcomes were also evaluated. SEARCH STRATEGY: Electronic searches of Cochrane Controlled Trials Register (Cochrane Library 2001, issue 4), MEDLINE (1966 - October 2001), EMBASE (1980 - December 2001), and CINHAL (1982 - December 2001) were undertaken. Du Pont Pharmaceutical and Ivax Corporation were contacted for information regarding unpublished trials. The reference lists of the obtained papers were also examined. SELECTION CRITERIA: All relevant randomised controlled trials (RCTs) and controlled clinical trials (CCTs) were included. Participants were people with alcohol dependence. Naltrexone (NTX), nalmefene (NMF) and other opioid antagonists with/without other biological or psychosocial treatments were examined. Four primary outcomes of interest were number of patients who return to drinking, percentage or number of drinking days, number of standard drinks of alcohol and amount of alcohol consumed. A number of secondary outcomes were also considered. DATA COLLECTION AND ANALYSIS: Two reviewers evaluated and extracted the data independently. The dichotomous data were extracted on an intention-to-treat basis. The Relative Risk with the 95% confidence interval was used to assess the dichotomous data. Weighted (or Standardised) Mean Difference with 95% confidence interval was used to assess the continuous data. MAIN RESULTS: The review included 19 RCTs or CCTs presented in 26 articles. In comparison to placebo, two of four short-term primary outcomes were significantly in favour of NTX. Those were number of patients who return to drinking (61% in NTX group vs 69% in placebo group) [RR (95% CI) = 0.88 (0.80 to 0.98), NNT = 14] and percentage or number of drinking days [WMD (95% CI) = -4.52 (-5.29 to -3.75)]. However, the short-term discontinuation rates were high and not different between NTX and placebo groups [RR (95% CI) = 0.96 (0.81 to 1.13)]. No medium-term outcomes of NTX and placebo groups showed any significant difference after the completion of NTX treatment for three to six months. However, those who were regularly treated with NTX treatment in both short and medium terms consumed smaller amounts of alcohol than placebo-treated patients. Because of the small sample sizes, there were few significant differences for other comparisons. REVIEWER'S CONCLUSIONS: NTX at the dose of 50 mg/day is effective for alcohol dependence in short-term treatment. The optimal duration of NTX treatment may be longer than 3 months. The evidence so far may be too little to support the superiority of NTX to acamprosate and the inferiority of NTX to disulfiram. NTX treatment should be concurrently given with a psychosocial intervention. Other patterns of NTX administration should not be used at present, e.g., a dose of three times a week, combined NTX with other biological treatments. NMF has no role for the treatment of alcohol dependence in clinical practice. Randomised, double-blind, placebo controlled trials of NTX treatment in patients with alcohol dependence are still needed. Some issues should be concerned in further studies. Firstly, further trials should be conducted in larger sample sizes and over longer periods of time. Secondly, other than the outcomes relevant to alcohol use, some important outcomes should also be measured, e.g., functioning, health-related quality of life, economic cost. Thirdly, the comparisons between NTX and other treatments for alcohol dependence, both biological and psychosocial, should be investigated. Fourthly, combined treatments of NTX and other biological treatments for alcohol dependence may be in issue of interest. Lastly, high discontinuation rate in both treatment and control groups should be concerned. PMID- 12076426 TI - Interventions for preventing obesity in children. AB - BACKGROUND: The prevalence of obesity and overweight is increasing in both adult and child populations throughout the world. Obesity in children impacts on their health in both the short and longer term, and obesity prevention is an international public health priority. However, the efficacy of prevention strategies is poorly understood. OBJECTIVES: To assess the effectiveness of educational, health promotion and/or psychological/family/behavioural therapy/counselling/management interventions that focussed on diet, physical activity and/or lifestyle and social support, and were designed to prevent obesity in childhood. SEARCH STRATEGY: The following databases were searched: MEDLINE, Psyclit, EMBASE, Science Citation Index, Social Science Citation Index, CINAHL, Cochrane Controlled Trials Register (CCTR) and the Cochrane Heart Group's specialised register from 1985 to July 2001. Non English language papers were included. Experts were contacted to seek additional references or unpublished studies. SELECTION CRITERIA: RCTs and non-randomised trials with concurrent control group that observed participants for a minimum of three months were included DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Ten studies were included; seven were long-term (children observed for at least one year), three were shorter term (at least 3 months). Eight were school/nursery-based interventions, one was a community-based intervention targeting low-income African-American families, and one was a family-based intervention that targeted non-obese children of obese parents. The studies included were diverse in terms of study design and quality, target population, theoretical underpinning of intervention approach, and outcome measures, so it was not possible to combine study findings using statistical methods. Three of the four long-term studies that combined dietary education and physical activity interventions resulted in no difference in overweight, whereas one study reported an improvement in favour of the intervention group. In two studies of dietary education alone, a multimedia action strategy appeared to be effective but other strategies did not. The one long term study that only focussed on physical activity resulted in a slightly greater reduction in overweight in favour of the intervention group, as did two short term studies of physical activity. REVIEWER'S CONCLUSIONS: There is limited high quality data on the effectiveness of obesity prevention programs and no generalisable conclusions can be drawn. However, concentration on strategies that encourage reduction in sedentary behaviours and increase in physical activity may be fruitful. The need for well-designed studies that examine a range of interventions remains a priority, although a number of important studies are underway. PMID- 12076427 TI - Carbamazepine versus phenytoin monotherapy for epilepsy. AB - BACKGROUND: Worldwide, carbamazepine and phenytoin are commonly used antiepileptic drugs. This review summarizes evidence from randomized controlled trials in which these two drugs have been compared. OBJECTIVES: To review the best evidence comparing carbamazepine and phenytoin when used as monotherapy in subjects with partial onset seizures, or generalized onset tonic-clonic seizures with or without other generalized seizure types. SEARCH STRATEGY: We searched: (a) the trial register of the Cochrane Epilepsy Group; (b) The Cochrane Controlled Trials Register (Cochrane Library Issue 4, 2001); (c) MEDLINE 1966 2001. In addition we hand searched relevant journals and contacted the pharmaceutical industry and researchers in the field to seek any ongoing or unpublished studies. SELECTION CRITERIA: Randomized controlled trials in children or adults with partial onset seizures or generalized onset tonic-clonic seizures. Trials must have included a comparison of carbamazepine monotherapy with phenytoin monotherapy. DATA COLLECTION AND ANALYSIS: This was an individual patient data review. Outcomes were time to (a) withdrawal of allocated treatment, (b) 12 month remission, (c) six month remission, and (d) first seizure post randomization. Data were analysed using a stratified logrank analysis with results expressed as hazard ratios (HR) and 95% confidence intervals (95% CI), where a HR>1 indicates an event is more likely on phenytoin. MAIN RESULTS: Individual patient data are available for 551 participants from three trials, representing 63% of the participants recruited into the nine trials that met our inclusion criteria. By convention, for the outcomes time to six and 12 month remission HR>1 indicates a clinical advantage for phenytoin, whilst for time to withdrawal and first seizure HR>1 indicates a clinical advantage for carbamazepine. Results (HR (95% CI)) were: (i) time to withdrawal of allocated treatment 0.97(0.74 to 1.28), (ii) time to 12 month remission 1.00(0.78 to 1.29), (iii) time to six month remission 1.10(0.87 to 1.39), (iv) time to first seizure 0.91(0.74 to 1.12). The results suggest no overall difference between carbamazepine and phenytoin for these outcomes. REVIEWER'S CONCLUSIONS: We have not found evidence that a significant difference exists between carbamazepine and phenytoin for the outcomes examined in this review. Confidence intervals are wide and the possibility of important differences existing has not been excluded. PMID- 12076428 TI - Psychological therapies for sickle cell disease and pain. AB - BACKGROUND: Sickle cell disease comprises a group of genetic blood disorders, and occurs when the sickle haemoglobin gene is inherited from both parents. The effects of the condition are: varying degrees of anaemia which if severe reduce the capacity for mobility; predisposition to obstruction of small blood capillaries causing pain in muscle and bone known as "crises"; damage to major organs such as the spleen, liver, kidneys, and lungs; and increased vulnerability to severe infections. There are both medical and non-medical complications, and treatment is usually symptomatic and palliative in nature. Psychological intervention for individuals with sickle cell disease seems viable in complementing current medical treatment, and studies examining their efficacy appear to have also yielded encouraging results. OBJECTIVES: To examine the evidence that in patients with sickle cell disease, psychological treatment improves the ability to cope with the condition. SEARCH STRATEGY: The Cochrane Cystic Fibrosis and Genetic Disorders Group specialist trials register which comprises references from comprehensive electronic database searches. Also, hand searching relevant journals, hand searching abstract books of conference proceedings, and searches on the Internet were performed. Date of the most recent search of the Group's specialised register: January 2001. SELECTION CRITERIA: All randomised or quasi-randomised controlled trials comparing the use of psychological intervention to no (psychological) intervention in patients with sickle cell disease. DATA COLLECTION AND ANALYSIS: Both reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Five studies were identified in the initial search, of which three studies, with a total of 158 patients were eligible for inclusion in the review. Published data reveal that family education and cognitive behavioural therapy can help patients cope with sickle cell disease. REVIEWER'S CONCLUSIONS: Patient education programmes improve knowledge and attitudes of patients with sickle cell disease. There is as yet however, insufficient evidence to demonstrate the role of other psychological therapies. This systematic review has clearly identified the need for well designed, adequately powered, multicentre randomised controlled trials assessing the effectiveness of specific intervention in sickle cell disease. PMID- 12076429 TI - Massage for low back pain. AB - BACKGROUND: Low-back pain is one of the most common and costly musculoskeletal problems in modern society. Proponents of massage therapy claim it can minimize pain and disability, and speed return to normal function. OBJECTIVES: To assess the effects of massage therapy for non-specific low-back pain. SEARCH STRATEGY: We searched Medline, Embase, Cochrane Controlled Trials Register, HealthSTAR, CINAHL and Dissertation abstracts from their beginning to May 2001 with no language restrictions. References in the included studies and in reviews of the literature were screened. Contact with content experts and massage associations was also made. SELECTION CRITERIA: The studies had to be randomized or quasi randomized trials investigating the use of any type of massage (using the hands or a mechanical device) as a treatment for non-specific low-back pain. DATA COLLECTION AND ANALYSIS: Two reviewers blinded to authors, journal and institutions selected the studies, assessed the methodological quality using the criteria recommended by the Cochrane Back Review Group, and extracted the data using standardized forms. The studies were analysed in a qualitative way due to heterogeneity of population, massage technique, comparison groups, timing and type of outcome measured. MAIN RESULTS: Nine publications reporting on eight randomized trials were included. Three had low and five had high methodological quality scores. One study was published in German and the rest in English. Massage was compared to an inert treatment (sham laser) in one study that showed that massage was superior, especially if given in combination with exercises and education. In the other seven studies, massage was compared to different active treatments. They showed that massage was inferior to manipulation and TENS; massage was equal to corsets and exercises; and massage was superior to relaxation therapy, acupuncture and self-care education. The beneficial effects of massage in patients with chronic low-back pain lasted at least one year after the end of the treatment. One study comparing two different techniques of massage concluded in favour of acupuncture massage over classic (Swedish) massage. REVIEWER'S CONCLUSIONS: Massage might be beneficial for patients with subacute and chronic non-specific low-back pain, especially when combined with exercises and education. The evidence suggest that acupuncture massage is more effective than classic massage, but this need confirmation. More studies are needed to confirm these conclusions and to assess the impact of massage on return-to-work, and to measure longer term effects to determine cost-effectiveness of massage as an intervention for low-back pain. PMID- 12076430 TI - Surgical versus non-surgical management of pleural empyema. AB - BACKGROUND: Pleural empyema is a collection of pus between the lungs and the chest wall. Approximately 50% of cases complicate pneumonia. There are a variety of treatment options ranging from intravenous antibiotics alone to open thoracotomy and debridement, depending in part on the stage of the empyema and the severity. The condition changes with time, becoming loculated and more difficult to drain. There is much debate about the most appropriate therapy particularly with the advent of new treatments such as fibrinolytic enzymes (e.g. streptokinase, urokinase) and video-assisted thoracoscopic surgery (VATS). OBJECTIVES: To determine which is more effective for the management of empyema: surgical (e.g. thoracoscopy, thoracotomy) or non-surgical techniques (e.g. thoracocentesis, chest tube drainage) and to establish whether there is an optimum time for intervention. SEARCH STRATEGY: The Cochrane Controlled Trials Register and DARE database were searched in addition to the Cochrane Acute Respiratory Infections Group's own register of trials. A specialised topic search with no language restrictions was used to search MEDLINE and EMBASE using Silverplatter. Bibliographies and the reference lists of identified studies and review articles were handsearched. Personal communication with authors and experts in the field is ongoing. SELECTION CRITERIA: Randomised controlled trials (RCTs) of surgical techniques versus non-surgical approaches for treatment of pus in the pleural cavity in children and adults but not neonates. Studies of empyema associated with tuberculosis or malignancy were excluded. DATA COLLECTION AND ANALYSIS: Trial quality was assessed using Jadad criteria as recommended by the ARI group (Jadad 1996). The primary outcomes were death or resolution of the empyema. Secondary outcomes addressed length of time chest tubes were required, pain, hospital stay and any complications. MAIN RESULTS: Only one small randomised study was identified which met the inclusion criteria. It was conducted in a university thoracic surgery department. There were some methodological quality considerations which cast some doubt on validity ( patient selection, unclear allocation concealment and outcome assessor blinding) and it scored 'B' overall (Jadad score 3). The main results of the study were that when compared with chest tube drainage combined with streptokinase, the video-assisted thoracoscopic surgery (VATS) group had a significantly higher primary treatment success and spent less time in hospital. Each group suffered one mortality. REVIEWER'S CONCLUSIONS: It would appear that for large, loculated pleural empyemas VATS is superior to chest tube drainage with streptokinase in terms of duration of chest tubes and hospital stay. However there are questions about validity and the study is also too small to draw conclusions. There are risks of complications (associated with all treatments) which may not apparent with small numbers. VATS is performed under general anaesthetic and one lung ventilation. Fibrinolytics are also associated with side effects. Further larger multicentre studies need to be conducted. PMID- 12076431 TI - Opioid antagonists with minimal sedation for opioid withdrawal. AB - BACKGROUND: Managed withdrawal (detoxification) is necessary prior to drug-free treatment. It may also represent the end point of long-term opioid replacement treatment such as methadone maintenance. OBJECTIVES: To assess the effectiveness of interventions involving opioid antagonists to induce withdrawal, in combination with medication to ameliorate symptoms but with minimal sedation. SEARCH STRATEGY: Multiple databases were searched using a strategy designed to retrieve references broadly addressing opioid withdrawal. Reference lists of retrieved studies, reviews and conference abstracts were handsearched. SELECTION CRITERIA: Randomised or quasi-randomised controlled clinical trials or prospective controlled cohort studies that compared antagonist-induced (conscious) withdrawal with other approaches to modify the signs and symptoms of withdrawal in opioid-dependent participants. DATA COLLECTION AND ANALYSIS: One reviewer assessed potentially relevant studies for inclusion and undertook data extraction. All inclusion decisions and the overall process were confirmed by consultation between all reviewers. MAIN RESULTS: Ten studies (5 randomised and 5 non-randomised controlled trials), involving 770 participants, met the inclusion criteria for the review. Treatment regimes using opioid antagonists to induce withdrawal, with minimal sedation, varied in a number of aspects preventing description of a "standard" approach. Antagonist-induced withdrawal is associated with similar or less overall severity than withdrawal managed primarily with an alpha2 adrenergic agonist. This is probably because of earlier resolution of withdrawal. Peak severity is likely to be higher with antagonist-induced withdrawal and require the use of additional adjunct medications. Withdrawal from methadone may be more severe than withdrawal from heroin, but data are limited. Antagonist-induced withdrawal appears to be associated with somewhat higher rates of completion of withdrawal and achievement of maintenance doses of naltrexone but there were insufficient data for statistical analyses. The benefit of higher rates of completion of withdrawal is lessened by apparently low rates of retention in subsequent naltrexone maintenance treatment. REVIEWER'S CONCLUSIONS: The use of opioid antagonists combined with alpha2 adrenergic agonists is feasible and probably increases the likelihood of transfer to naltrexone compared to withdrawal managed primarily with an adrenergic agonist. A high level of monitoring and support is desirable for several hours following administration of opioid antagonists because of the possibility of vomiting, diarrhoea and delirium. Further research is required to confirm the relative effectiveness of antagonist-induced regimes, as well as variables influencing the severity of withdrawal, adverse effects, the most effective antagonist-based treatment regime, and approaches that might increase retention in subsequent naltrexone maintenance treatment. PMID- 12076433 TI - Carbamazepine for cocaine dependence. AB - BACKGROUND: Cocaine dependence has become a substantial public health problem, developing a significant number of medical, psychological and social problems, including the spread of infectious diseases (e.g. AIDS, hepatitis and tuberculosis), crime, violence and neonatal drug exposure. Although there is no consensus regarding how to treat cocaine dependence, effective pharmacotherapy has a potentially major role to play as part of a broader treatment milieu. The anti-convulsant carbamazepine, a tricyclic medication that is widely used to treat a variety of neurological and psychiatric disorders, has also been used for treatment of cocaine dependence, although its effectiveness has not been established. OBJECTIVES: To determine whether carbamazepine (CBZ) is effective for the treatment of cocaine dependence. SEARCH STRATEGY: We searched: the Cochrane Controlled Trials Register (Cochrane Library issue 1, 1999), MEDLINE (from 1966 - October 1997), EMBASE (from 1980 - October 1997), PsycLIT (from 1974 - July 1997), Biological Abstracts and LILACS (from 1982 - 1997); scan of reference list of relevant articles; personal communication; conference abstracts; unpublished trials from pharmaceutical industry; book chapters on treatment of cocaine dependence. SELECTION CRITERIA: The inclusion criteria for all randomised controlled trials were that they should focus on the use of carbamazepine drugs versus placebo on the treatment of cocaine dependence. Trials including patients with additional diagnosis such as opiate dependence were also eligible. DATA COLLECTION AND ANALYSIS: The reviewers extracted the data independently and Odds Ratios, weighted mean difference and number needed to treat were estimated. Qualitative assessments of the methodology of eligible studies were carried out using validated checklists. The reviewers assumed that people who died or dropped out had no improvement and tested the sensitivity of the final results to this assumption. Where possible analysis was carried out according to the "intention to treat" principles. MAIN RESULTS: 5 studies were included in the review, with 455 people randomised. No differences were found regarding positive urine sample for cocaine metabolites. Scores on Spielberg State Anxiety Inventory slightly favoured carbamazepine, but didn't reach statistical significance. Dropouts were high in both groups up to 70% in the placebo group. Less dropout occurred in the carbamazepine group (RR 0.87 95%CI 0.71-1.06). When no retention in treatment was due to side effects no differences were found. The number of participants presenting at least one side effect, reported in Kranzler (Kranzler 1995), was higher in the carbamazepine group (RR 4.33 95% CI 1.45-12.91). REVIEWER'S CONCLUSIONS: There is no current evidence supporting the clinical use of CBZ in the treatment of cocaine dependence. Larger randomised investigation must be considered taking into account that these time consuming efforts should be reserved for medications showing more relevant and promising evidence. PMID- 12076432 TI - Opioid antagonists under heavy sedation or anaesthesia for opioid withdrawal. AB - BACKGROUND: Withdrawal (detoxification) is necessary prior to drug-free treatment. It may also represent the end point of long-term opioid replacement treatment such as methadone maintenance. The availability of managed withdrawal is essential to an effective treatment system. OBJECTIVES: To assess the effectiveness of interventions involving the administration of opioid antagonists to induce opioid withdrawal with concomitant heavy sedation or anaesthesia. SEARCH STRATEGY: Multiple electronic databases (including MEDLINE, EMBASE, PsycLIT, Australian Medical Index, Cochrane Controlled Trials Register, and CINAHL) were systematically searched. Reference lists of retrieved studies, reviews and conference abstracts were handsearched. SELECTION CRITERIA: Randomised or quasi-randomised controlled trials that compared the administration of opioid antagonists under heavy sedation or anaesthesia with another form of treatment. DATA COLLECTION AND ANALYSIS: One reviewer assessed studies for inclusion and undertook data extraction. Inclusion decisions and the overall process were confirmed by consultation between all three reviewers. MAIN RESULTS: As yet, no studies have been published comparing treatment regimes involving the administration of opioid antagonists under heavy sedation or anaesthesia with other approaches to detoxification. Treatment regimes for the studies considered for this review varied in the opioid antagonist used, the dose and mode of administration, the anaesthetic agent, duration of anaesthesia and adjunct medications employed. More detailed monitoring of withdrawal is required before any conclusions can be drawn as to what comprises a typical pattern of withdrawal and what factors might influence the pattern. There is only very limited information on referral to ongoing treatment, and relapse to opioid use. Together with the lack of adequate comparisons, this makes it impossible to draw any conclusions about the long-term effectiveness, or the cost-effectiveness, of withdrawal induced by opioid antagonists under heavy sedation or anaesthesia. REVIEWER'S CONCLUSIONS: Considerably more research evidence will be needed before any conclusions can be drawn regarding the effectiveness of managing withdrawal by administration of opioid antagonists under heavy sedation or anaesthesia. The risk of vomiting during sedation, respiratory depression and cardiac irregularities point to the approach being limited to facilities equipped for intubation, assisted ventilation and a high level of monitoring, and with the capacity to respond to adverse events that might occur. The approach must be regarded as experimental with both risks and benefits remaining uncertain. PMID- 12076434 TI - Buprenorphine for the management of opioid withdrawal. AB - BACKGROUND: Managed withdrawal (detoxification) is a necessary step prior to drug free treatment. It may also represent the end point of long-term opioid replacement treatment such as methadone maintenance. The availability of managed withdrawal is essential to an effective treatment system. OBJECTIVES: To assess the effectiveness of interventions involving the short-term use of buprenorphine to manage the acute phase of opioid withdrawal. SEARCH STRATEGY: Multiple electronic databases were searched using a strategy designed to retrieve references broadly addressing the management of opioid withdrawal. Reference lists of retrieved studies, reviews and conference abstracts were handsearched. SELECTION CRITERIA: Randomised or quasi-randomised controlled clinical trials or prospective controlled cohort studies that compared different buprenorphine regimes, or that compared buprenorphine with another form of treatment (or placebo) to modify the signs and symptoms of withdrawal in participants who were primarily opioid dependent. DATA COLLECTION AND ANALYSIS: Potentially relevant studies were assessed for inclusion by one reviewer. Inclusion decisions were confirmed by consultation between reviewers. One reviewer undertook data extraction with the process confirmed by consultation between all three reviewers. MAIN RESULTS: Six studies (5 RCTs and 1 controlled prospective study), involving 357 participants, met the criteria for inclusion in the review. Four studies compared buprenorphine with clonidine. All found withdrawal to be less severe in the buprenorphine treatment group. In three of these studies all participants were withdrawing from heroin. Participants in one study were withdrawing from methadone (10mg/day). Aches, restlessness, yawning, mydriasis, tremor, insomnia, nausea and mild anxiety were reported as being experienced by some participants. Rates of completion of withdrawal ranged from 65% to 100%. None of the studies included in the review reported adverse effects. However a single-group study which therefore did not meet the inclusion criteria, reported the occurrence in some participants of headaches, sedation, nausea, constipation, anxiety, dizziness and itchiness, particularly in the first 2-3 days of treatment. In one of the six studies, and in two studies that did not meet the inclusion criteria, treatment was provided on an outpatient basis. REVIEWER'S CONCLUSIONS: Buprenorphine has potential as a medication to ameliorate the signs and symptoms of withdrawal from heroin, and possibly methadone, but many aspects of treatment protocol and relative effectiveness need to be investigated further. PMID- 12076435 TI - Radiotherapy versus open surgery versus endolaryngeal surgery (with or without laser) for early laryngeal squamous cell cancer. AB - BACKGROUND: Radiotherapy, open surgery and endolaryngeal excision (with or without laser) are all accepted modalities of treatment for early stage glottic cancer. Case series suggest that they confer similar survival advantage. Opinions on optimal therapy vary across disciplines and between countries. OBJECTIVES: To compare the effectiveness of open surgery, endolaryngeal excision (with or without laser) and radiotherapy in the management of early glottic laryngeal cancer SEARCH STRATEGY: Electronic search of MEDLINE (from 1966 to October 2000), EMBASE (from 1980 to October 2000), CINAHL (from 1982 to October 2000) and CancerLit (from 1963 to October 2000) databases and the Cochrane Controlled Trials Register. SELECTION CRITERIA: Randomised controlled trials (RCT) comparing open surgery, endolaryngeal resection and/or radiotherapy DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed RCTs identified from the electronic searches for eligibility and methodological quality. All authors of the review discussed the results of these assessments. MAIN RESULTS: Only one RCT was identified which compared open surgery and radiotherapy among a substantial number of patients with early glottic laryngeal cancer. REVIEWER'S CONCLUSIONS: There is currently insufficient evidence to guide management decisions on the most effective treatment. Interpretation of the only large scale RCT comparing open surgery and radiotherapy in patients with early glottic cancer is limited because of concerns about the adequacy of treatment regimens and deficiencies in the reporting of the study design and analysis. Endolaryngeal resection of early glottic tumours is becoming more common and a well designed multicentre RCT is warranted. PMID- 12076436 TI - Nystatin prophylaxis and treatment in severely immunodepressed patients. AB - BACKGROUND: Nystatin is sometimes used prophylactically in patients with severe immunodeficiency or in the treatment of fungal infection in such patients, although the effect seems to be equivocal. OBJECTIVES: To study whether nystatin decreases morbidity and mortality when given prophylactically or therapeutically to patients with severe immunodeficiency. SEARCH STRATEGY: MEDLINE and The Cochrane Library using a comprehensive search strategy, date of last search November 2001. Contacted industry and scanned reference lists. SELECTION CRITERIA: Randomised trials comparing nystatin with placebo, an untreated control group, fluconazole or amphotericin B. DATA COLLECTION AND ANALYSIS: Data on mortality, invasive fungal infection and colonisation were extracted by both authors independently. The outcomes were weighted by the inverse variance. A random effects model was used unless p>0.10 for the test of heterogeneity. MAIN RESULTS: We included 12 trials (1,464 patients). The drugs were given prophylactically in ten trials and as treatment in two. Seven trials were in acute leukaemia, two in cancer, one in liver transplant patients, one in critically ill surgical and trauma patients, and one in AIDS patients. Nystatin had been compared with placebo in three trials and with fluconazole in nine; the dose varied from 1.5 MIE to 72 MIE daily. The effect of nystatin was similar to that of placebo on fungal colonisation (relative risk 0.85, 95% confidence interval 0.65 to 1.13). There was no statistically significant difference between fluconazole and nystatin on mortality (relative risk 0.76, 0.49 to 1.18) whereas fluconazole was more effective in preventing invasive fungal infection (relative risk 0.37, 0.15 to 0.91) and colonisation (relative risk 0.49, 0.34 to 0.70). The results were very similar if the three studies which were not performed in cancer patients were excluded. REVIEWER'S CONCLUSIONS: Nystatin cannot be recommended for prophylaxis or treatment of Candida infections in immunodepressed patients. PMID- 12076437 TI - Transfusion thresholds and other strategies for guiding allogeneic red blood cell transfusion. AB - BACKGROUND: Most clinical practice guidelines recommend restrictive red cell transfusion practices with the goal of minimising exposure to allogeneic blood (from an unrelated donor). The purpose of this review is to compare clinical outcomes in patients randomised to restrictive versus liberal transfusion thresholds (triggers). OBJECTIVES: To examine the evidence on the effect of transfusion thresholds, on the use of allogeneic and/or autologous blood, and the evidence for any effect on clinical outcomes. SEARCH STRATEGY: Trials were identified by: computer searches of OVID Medline (1966 to December 2000), Current Contents (1993 to Week 48 2000), and the Cochrane Controlled Trials Register (2000 Issue 4). References in identified trials and review articles were checked and authors contacted to identify any additional studies. SELECTION CRITERIA: Controlled trials in which patients were randomised to an intervention group or to a control group. Trials were included where the intervention groups were assigned on the basis of a clear transfusion "trigger", described as a haemoglobin (Hb) or haematocrit (Hct) level below which a RBC transfusion was to be administered. DATA COLLECTION AND ANALYSIS: Trial quality was assessed using criteria proposed by Schulz et al. (1995). Relative risks of requiring allogeneic blood transfusion, transfused blood volumes and other clinical outcomes were pooled across trials using a random effects model. MAIN RESULTS: Ten trials were identified that reported outcomes for a total of 1780 patients. Restrictive transfusion strategies reduced the risk of receiving a red blood cell (RBC) transfusion by a relative 42% (RR=0.58: 95%CI=0.47,0.71). This equates to an average absolute risk reduction (ARR) of 40% (95%CI=24% to 56%). The volume of RBCs transfused was reduced on average by 0.93 units (95%CI=0.36,1.5 units). However, heterogeneity between these trials was statistically significant (p<0.00001) for these outcomes. Mortality, rates of cardiac events, morbidity, and length of hospital stay were unaffected. Trials were of poor methodological quality. REVIEWER'S CONCLUSIONS: The limited published evidence supports the use of restrictive transfusion triggers in patients who are free of serious cardiac disease. However, most of the data on clinical outcomes were generated by a single trial. The effects of conservative transfusion triggers on functional status, morbidity and mortality, particularly in patients with cardiac disease, need to be tested in further large clinical trials. In countries with inadequate screening of donor blood the data may constitute a stronger basis for avoiding transfusion with allogeneic red cells. PMID- 12076438 TI - Bisphosphonates for the relief of pain secondary to bone metastases. AB - BACKGROUND: Bisphosphonates form part of standard therapy for hypercalcemia and the prevention of skeletal events in some cancers. However, the role of bisphosphonates in pain relief for bony metastases remains uncertain. OBJECTIVES: To determine the effectiveness of bisphosphonates for the relief of pain from bone metastases. SEARCH STRATEGY: MEDLINE (1966-1999), EMBASE (1980-1999), CancerLit (1966-1999), the Cochrane library (Issue 1, 2000) and the Oxford Pain Database were searched using the strategy devised by the Cochrane Pain, Palliative and Supportive Care Group with additional terms 'diphosphonate', 'bisphosphonate', 'multiple myeloma' and 'bone neoplasms'. (Last search: January 2000). SELECTION CRITERIA: Randomized trials of bisphosphonates compared with open, blinded, or different doses/types of bisphosphonates in cancer patients were included where pain and/or analgesic consumption were outcome measures. Studies where pain was reported only by observers were excluded. DATA COLLECTION AND ANALYSIS: Article eligibility, quality assessment and data extraction were undertaken by both reviewers. The proportions of patients with pain relief at 4, 8 and 12 weeks were assessed. The proportion of patients with analgesic reduction, the mean pain score, mean analgesic consumption, adverse drug reactions, and quality of life data were compared as secondary outcomes. MAIN RESULTS: Thirty randomized controlled studies (21 blinded, four open and five active control) with a total of 3682 subjects were included. For each outcome, there were few studies with available data. For the proportion of patients with pain relief (eight studies) pooled data showed benefits for the treatment group, with an NNT at 4 weeks of 11[95% CI 6-36] and at 12 weeks of 7 [95% CI 5-12]. In terms of adverse drug reactions, the NNH was 16 [95% CI 12-27] for discontinuation of therapy. Nausea and vomiting were reported in 24 studies with a non-significant trend for greater risk in the treatment group. One study showed a small improvement in quality of life for the treatment group at 4 weeks. The small number of studies in each subgroup with relevant data limited our ability to explore the most effective bisphosphonates and their relative effectiveness for different primary neoplasms. REVIEWER'S CONCLUSIONS: There is evidence to support the effectiveness of bisphosphonates in providing some pain relief for bone metastases. There is insufficient evidence to recommend bisphosphonates for immediate effect; as first line therapy; to define the most effective bisphosphonates or their relative effectiveness for different primary neoplasms. Bisphosphonates should be considered where analgesics and/or radiotherapy are inadequate for the management of painful bone metastases. PMID- 12076439 TI - Cleavage stage versus blastocyst stage embryo transfer in assisted conception. AB - BACKGROUND: Despite numerous advances in the field of in vitro fertilisation (IVF), many of the widely applied embryo culture techniques and resulting implantation rates have remained relatively unchanged since the first treatment was performed in the mid 1970's. Recent advances in the understanding of nutrient requirements of embryos, have led to a renaissance of extending their culture from the standard procedure of 2-3 days (early cleavage embryo transfer) to 5-6 days (blastocyst culture). The rationale for blastocyst culture is to improve the synchronicity of uterine and embryonic development and provide a mechanism for self-selection of viable embryos. Numerous reports on the clinical benefits of blastocyst culture have led to the worldwide introduction of this technique, despite a deficiency of conclusive evidence to do so. OBJECTIVES: Primary: To determine if blastocyst stage embryo transfers (ET's) result in higher success rates, than cleavage stage embryo transfers. Secondary: To assess the overall embryo utilisation rate of both techniques. SEARCH STRATEGY: Electronic searches of the Cochrane Menstrual Disorders and Subfertility Group specialised register of controlled trials, CCTR, MEDLINE, EMBASE, and Bio extracts were performed to identify relevant randomised controlled trials (RCTs). Attempts were also made to identify trials from the National Research Register, the Clinical Trial Register and the citation lists of review articles and included trials. The first or corresponding author of each included trial was also contacted for additional information. SELECTION CRITERIA: Trials were included if they were randomised and compared the effectiveness of early cleavage versus blastocyst stage transfers. DATA COLLECTION AND ANALYSIS: Of the 29 trials that were identified, ten trials met the inclusion criteria and were reviewed. Primary outcomes were rates of; live birth, clinical pregnancy and implantation per woman. Secondary outcomes were rates of; miscarriage, monozygotic twinning, embryo freezing, embryo utilisation, cancellation, multiple pregnancy and high order pregnancy and per cycle data. Quality assessment and data extraction were performed independently by two reviewers. Meta analysis was performed using odds ratios for dichotomous outcomes and weighted mean differences for continuous outcomes. MAIN RESULTS: There was no significant difference between the two treatment groups in live birth rate, although this was reported by only one quasi-random trial (Peto OR 1.59, 95% CI 0.80, 3.15). There was also no evidence of a difference in pregnancy rate (both overall and subgroups) between the two groups for pregnancy rate per couple randomised (4 RCTs: Peto OR 0.86, 95% CI 0.57, 1.29). There was also no suggestion of an overall difference in implantation rates per embryo's transferred although it was impossible to calculate valid confidence intervals from published data (Day 2/3 17.1% vs Day 5/6 18.9%). The subgroup of sequential media trials suggested higher implantation rate for blastocyst transfer (Day 2/3 22.6% vs Day 5/6 32%). The miscarriage rate was no different between the two groups (1 RCT, Peto OR 1.66, 95% CI 0.41, 6.81). The RCTs reporting embryo freezing showed no difference (Peto OR 1.71, 95% CI 1.00, 2.94), however the two quasi-random trials showed a significant difference in favour of the Day 2/3 group (Peto OR 2.99, 95% CI 1.88, 4.75). Embryo transfer cancellation rates were significantly higher in the Day 5/6 group (5 RCTs: Peto OR 0.57, 95% CI 0.40, 0.83). There was no significant difference in the rate of multiple pregnancies or the rate of high order pregnancies (3 RCTs, Peto OR 0.58, 95% CI 0.30, 1.12)(2 RCTs, Peto OR 7.88, 95% CI 0.49, 126.30 respectively). REVIEWER'S CONCLUSIONS: Overall this review of the best available evidence based on data from randomised controlled trials, suggests that to date little difference in the major outcome parameters has been demonstrated between early embryo transfer and blastocyst culture. Collectively, the increase in cancellation and the possible decrease in cryopreservation rates suggest that the routine practice of blastocyst culture should be offered to patients with caution. The subgroup of trials employing sequential media, did however demonstrate a substantial improvement in implantation rates and similar pregnancy rates, despite the transfer of less embryos. Whether this trend will culminate in convincing higher live birth rates per woman, has yet to be validated. PMID- 12076440 TI - Routine anticonvulsants for treating cerebral malaria. AB - BACKGROUND: Cerebral malaria is a common complication of Plasmodium falciparum infection, and kills over a million people every year. People with cerebral malaria become unconscious, and often have protracted convulsions. It is unclear whether giving anticonvulsant drugs routinely to people with cerebral malaria will improve the outcome of treatment and prevent death. OBJECTIVES: To evaluate the effect of routine anticonvulsant drugs in people with cerebral malaria. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group specialized trials register (November 2001), The Cochrane Controlled Trials Register (Issue 4, 2001), MEDLINE (1966 to November 2001), EMBASE (1988 to October 2001), LILACS (2001, 40a Edition CD-ROM), Science Citation Index (November 2001), African Index Medicus (1999), reference lists of articles, and research organizations. We also contacted the authors for addtional information. SELECTION CRITERIA: Randomized and quasi-randomized controlled trials of people with cerebral malaria. The trials compared anticonvulsant drugs started on admission to hospital with no anticonvulsant drug or placebo. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data from those trials eligible for inclusion. We assessed the methodological quality of the included trials by considering allocation sequence, concealment of allocation, blinding, and inclusion of all randomized participants. We used Review Manager (version 4.1) for the meta analysis and also explored possible sources of heterogeneity. MAIN RESULTS: Three trials with a total of 573 participants met the inclusion criteria. These trials all compared phenobarbitone with placebo or no treatment. In the two trials with adequate allocation concealment, death was more common in the anticonvulsant group (Relative Risk 2.0; 95% confidence interval 1.20 to 3.33; fixed effect model). In all three trials, phenobarbitone compared with placebo or no treatment was associated with fewer convulsions (Relative Risk 0.30; 95% confidence interval 0.19 to 0.45; fixed effect model). REVIEWER'S CONCLUSIONS: Routine phenobarbitone in cerebral malaria is associated with fewer convulsions but possibly more deaths. Further trials with adequate design, more participants, and different doses of anticonvulsant drugs are needed. PMID- 12076441 TI - LAAM maintenance vs methadone maintenance for heroin dependence. AB - BACKGROUND: LAAM and methadone are both full mu opiate agonists and have been shown to reduce dependence on heroin when given continuously under supervised dosing conditions. LAAM has a long duration of action requiring dosing every two or three days compared to methadone which requires daily dosing. LAAM is not as widely available internationally as methadone, and may be withdrawn from the market following ten cases of life-threatening cardiac arrhythmias and an association with QT prolongation. OBJECTIVES: To compare the efficacy and acceptability of LAAM maintenance with methadone maintenance in the treatment of heroin dependence. SEARCH STRATEGY: We searched MEDLINE (January 1966 to August 2000), PsycINFO (1887 to August 2000), EMBASE (January 1985 to August 2000), and the Cochrane Controlled Trials Register (Issue 2 2000). In addition we hand searched NIDA monographs until August 2000 and searched reference lists of articles. SELECTION CRITERIA: All randomised controlled trials, controlled clinical trials and controlled prospective studies comparing LAAM and methadone maintenance for the treatment of heroin dependence and measuring outcomes of efficacy or acceptability were included. DATA COLLECTION AND ANALYSIS: Data on retention in treatment, heroin use, side-effects and mortality were collected by two reviewers independently. A meta-analysis was performed using RevMan. Discrepancies were resolved by consensus. MAIN RESULTS: Eighteen studies, (15 RCTs, 3 Controlled prospective studies) met the inclusion criteria for the review. Three were excluded from the meta-analysis due to lack of data on retention, heroin use or mortality. Cessation of allocated medication (11 studies, 1473 participants) was greater with LAAM than with methadone, (RR 1.36, 95%CI 1.07-1.73, p=0.001, NNT=7.7 (or 8)). Non-abstinence was less with LAAM (5 studies, 983 participants; RR 0.81, 95%CI 0.72-0.91, p=0.0003, NNT=9.1 (or 10)). In 10 studies (1441 participants) there were 6 deaths from a range of causes, 5 in participants assigned to LAAM (RR 2.28 (95%CI 0.59-8.9, p=0.2). other relevant outcomes, such as quality of life and criminal activity could not be analysed because of lack of information in the primary studies. REVIEWER'S CONCLUSIONS: LAAM appears more effective than methadone at reducing heroin use. More LAAM patients than methadone ceased their allocated medication during the studies, but many transferred to methadone and so the significance of this is unclear. There was no difference in safety observed, although there was not enough evidence to comment on uncommon adverse events. PMID- 12076442 TI - Ribavirin with or without alpha interferon for chronic hepatitis C. AB - BACKGROUND: Hepatitis C is a major cause of liver-related morbidity and mortality. Ribavirin plus interferon combination therapy is presently considered the optimal treatment of interferon naive patients with chronic hepatitis C, but its role in relapsers and non-responders to previous interferon therapy is not established. OBJECTIVES: To assess the efficacy and safety of ribavirin alone or in combination with alpha interferon in interferon naive patients, relapsers, and non-responders with chronic hepatitis C. SEARCH STRATEGY: Eligible trials were identified through searches on electronic databases: The Cochrane Hepato-Biliary Group Controlled Trials Register (August 2001), The Cochrane Controlled Trials Register on The Cochrane Library Issue 3, 2001, MEDLINE (1966 - August 2001), and EMBASE (1985 - August 2001). Manual searches of bibliographies and journals were done as well as authors of trials and pharmaceutical companies producing ribavirin or interferon were contacted. SELECTION CRITERIA: We included all randomised trials comparing ribavirin with or without alpha interferon versus no intervention, placebo, or alpha interferon for chronic hepatitis C. DATA COLLECTION AND ANALYSIS: The primary outcome measures were the 'sustained' (six months after treatment) virological response, and morbidity plus mortality. The secondary outcome measures were the 'end of treatment' and 'sustained' biochemical response, the 'end of treatment' virologic response, histology, quality of life, and adverse events. MAIN RESULTS: We included eight trials in which 271 patients were randomised to ribavirin versus placebo or no intervention and 48 trials in which 6585 patients were randomised to interferon with or without ribavirin. Compared with placebo or no intervention, ribavirin monotherapy had no significant effect on the virological response or histology and only a transient effect on the biochemical response. Compared with interferon, combination therapy reduced the risk of not having a sustained virological response by 26% in naive patients (relative risk (RR) 0.74; 95% confidence interval (CI) 0.70-0.78), 33% in relapsers (RR 0.67; 95% CI 0.57 0.78), and 11% in non-responders (RR 0.89; 95% CI 0.83-0.96). There was no significant effect on morbidity plus mortality (Peto odds ratio 0.45; 95% CI 0.19 1.06). Irrespective of previous therapy, combination therapy significantly reduced the risk of not having a sustained biochemical response (RR 0.76; 95% CI 0.59-0.84) or improved histology (RR 0.67; 95% CI 0.56-0.81). Combination therapy also significantly increased the risk of treatment discontinuation (RR 1.28; 95% CI 1.07-1.52) and several types of adverse events. REVIEWER'S CONCLUSIONS: Combination therapy increased the number of naive patients, relapsers, and non responders with a sustained virological, biochemical, or histological response, but also the occurrence of adverse events. PMID- 12076443 TI - Calcium channel blockers for inhibiting preterm labour. AB - BACKGROUND: Preterm birth is a major contributor to perinatal mortality and morbidity and affects approximately six to seven per cent of births in developed countries. Tocolytics are drugs used to suppress uterine contractions. The most widely tested tocolytics are betamimetics. Although they have been shown to delay delivery, betamimetics have not been shown to improve perinatal outcome, and they have a high frequency of unpleasant and even fatal maternal side effects. There is growing interest in calcium channel blockers as a potentially effective and well tolerated form of tocolysis. OBJECTIVES: To assess the effects on maternal, fetal and neonatal outcomes of calcium channel blockers, administered as a tocolytic agent, to women in preterm labour. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group's specialised register of controlled trials, the Cochrane Controlled Trials Register (February 2002), MEDLINE, EMBASE, and Current Contents. We also contacted recognised experts and cross referenced relevant material. SELECTION CRITERIA: All published and unpublished randomised trials in which calcium channel blockers were used for tocolysis for women in labour between 20 and 36 weeks gestation. DATA COLLECTION AND ANALYSIS: Standard methods of the Cochrane Collaboration and the Cochrane Pregnancy and Childbirth Group were used. Evaluation of methodological quality and trial data extraction were undertaken independently by three authors. Additional information was sought to enable assessment of methodology and conduct of intention-to-treat analyses. Meta-analysis was conducted assessing the effects of calcium channel blockers compared with any other tocolytic agent. Results are presented using relative risk for categorical data and weighted mean difference for continuous data. MAIN RESULTS: Eleven randomised controlled trials involving 870 women were included. When compared with any other tocolytic agent (mainly betamimetics), calcium channel blockers reduced the number of women giving birth within 48 hours (relative risk (RR) 0.73; 95% confidence interval (CI) 0.54, 0.98) and within seven days (RR 0.76; 95% CI 0.59, 0.99). Calcium channel blockers also reduced the requirement for women to have treatment ceased for adverse drug reaction (RR 0.15; 95% CI 0.06, 0.43), the frequency of neonatal respiratory distress syndrome (RR 0.64; 95% CI 0.45, 0.91) and neonatal jaundice (RR 0.73; 95% CI 0.57, 0.93). REVIEWER'S CONCLUSIONS: When tocolysis is indicated for women in preterm labour, calcium channel blockers are preferable to betamimetic agents. Further research should address the effects of different dosage regimens and formulations of nifedipine on maternal and neonatal outcomes. PMID- 12076444 TI - Orthotic devices for treating patellofemoral pain syndrome. AB - BACKGROUND: Patellofemoral pain syndrome is a frequently reported condition in active adults. A wide variety of conservative treatment strategies have been described. As yet, no optimal strategy has been identified. Application of orthotic devices e.g. knee braces, knee straps, forms of taping of the knee, active training devices, knee sleeves and in-shoe orthotics to support the foot have been advocated to treat this condition. OBJECTIVES: To assess the effectiveness of foot and knee orthotics for treatment of patellofemoral pain syndrome. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register (December 2001), the Cochrane Controlled Trials register (Issue 2, 2000), MEDLINE (January 1966 to March 2000; EMBASE (January 1988 to March 2000); CINAHL (January 1982 to March 2000) and PEDro (up to March 2000). Relevant orthotic companies were contacted. SELECTION CRITERIA: All randomised and quasi-randomised trials comparing the effectiveness of knee or foot orthotics for treatment of patellofemoral pain syndrome were selected. Trials describing the use of orthotic devices in conjunction with operative treatment were excluded. DATA COLLECTION AND ANALYSIS: Three reviewers independently assessed methodological quality of the identified trials by use of a modified version of the Cochrane Musculoskeletal Injuries Group assessment tool, consisting 11 items. Two reviewers extracted data without blinding. Trialists were contacted to obtain missing data. MAIN RESULTS: Five trials involving 362 participants were included in this review. Five other trials await possible inclusion if further information can be obtained and one awaits further assessment. Due to clinical heterogeneity, we refrained from statistical pooling and conducted analysis by grading the strength of scientific evidence. The level of obtained research-based evidence was graded as limited as all trials were of low methodological quality. This limited research-based evidence showed the Protonics orthosis at six week follow-up was significantly more effective for decrease in pain (weighted mean difference (WMD) between groups 3.2; 95% confidence interval (CI) 2.8 to 3.6), functional improvement on the Kujala score (WMD 45.6; 95% CI 43.4 to 47.7) and change in patellofemoral congruence angle (WMD 17.2; 95% CI 14.1 to 20.3) when compared to no treatment. A comprehensive programme including tape application was significantly superior to a monitored exercise programme without tape application for decrease in worst pain (WMD 1.6; 95% CI 0.4 to 2.8) and usual pain (WMD 1.2; 95% CI 0.2 to 2.1), and clinical change and functional improvement questionnaire scores (WMD 10, 95% CI 2.07 to 17.93) at four weeks follow-up. The trials reported statistically significant differences in patient satisfaction after applied therapy (WMD 3.3; 95% CI 0.5 to 6.1) in favour of the McConnell regimen compared with the Coumans bandage at six weeks follow-up. REVIEWER'S CONCLUSIONS: The evidence from randomised controlled trials is currently too limited to draw definitive conclusions about the use of knee and foot orthotics for the treatment of patellofemoral pain. Future high quality trials in this field are warranted. PMID- 12076445 TI - Continuous distending pressure for respiratory distress syndrome in preterm infants. AB - BACKGROUND: Respiratory distress syndrome (RDS) is the single most important cause of morbidity and mortality in preterm infants (Greenough 1998, Bancalari 1992). Intermittent positive pressure ventilation (IPPV) with surfactant is the standard treatment for the condition. The major difficulty with IPPV is that it is invasive, resulting in airway and lung injury and contributing to the development of chronic lung disease. OBJECTIVES: In spontaneously breathing preterm infants with RDS, to determine if continuous distending pressure (CDP) reduces the need for IPPV and associated morbidity without adverse effects. SEARCH STRATEGY: The standard search strategy of the Neonatal Review group was used. This included searches of the Oxford Database of Perinatal Trials, Cochrane Controlled Trials Register (The Cochrane Library, Issue 1, 2002), MEDLINE (1966 January 2002), and EMBASE (1980-January 2002), previous reviews including cross references, abstracts, conference and symposia proceedings, expert informants, journal hand searching mainly in the English language. SELECTION CRITERIA: All trials using random or quasi-random allocation of preterm infants with RDS were eligible. Interventions were continuous distending pressure including continuous positive airway pressure (CPAP) by mask, nasal prong, nasopharyngeal tube, or endotracheal tube, or continuous negative pressure (CNP) via a chamber enclosing the thorax and lower body, compared with standard care. DATA COLLECTION AND ANALYSIS: Standard methods of the Cochrane Collaboration and its Neonatal Review Group were used, including independent assessment of trial quality and extraction of data by each author. MAIN RESULTS: CDP is associated with a lower rate of failed treatment (death or use of assisted ventilation) [summary RR 0.70 (0.55, 0.88), RD -0.22 (-0.35, -0.09), NNT 5 (3, 11)], overall mortality [summary RR 0.52 (0.32, 0.87), RD -0.15 (-0.26, -0.04), NNT 7 (4, 25)], and mortality in infants with birthweights above 1500 g [summary RR 0.24 (0.07, 0.84), RD -0.281 ( 0.483, -0.078), NNT 4 (2, 13)]. The use of CDP is associated with an increased rate of pneumothorax [summary RR 2.36 (1.25, 5.54), RD 0.14 (0.04, 0.23), NNH 7 (4, 24)]. REVIEWER'S CONCLUSIONS: In preterm infants with RDS the application of CDP either as CPAP or CNP is associated with benefits in terms of reduced respiratory failure and reduced mortality. CDP is associated with an increased rate of pneumothorax. The applicability of these results to current practice is difficult to assess, given the intensive care setting of the 1970s when four out of five of these trials were done. Where resources are limited, such as in developing countries, CPAP for RDS may have a clinical role. Further research is required to determine the best mode of administration and its role in modern intensive care settings PMID- 12076446 TI - Fluoride gels for preventing dental caries in children and adolescents. AB - BACKGROUND: Topically applied fluoride gels have been widely used as a caries preventive intervention in dental surgeries and school-based programs for over two decades. OBJECTIVES: To determine the effectiveness and safety of fluoride gels in the prevention of dental caries in children and to examine factors potentially modifying their effect. SEARCH STRATEGY: Multiple electronic database searches, reference lists of articles, journal handsearch, selected authors and manufacturers. SELECTION CRITERIA: Randomized or quasi-randomized controlled trials with blind outcome assessment, comparing fluoride gel with placebo or no treatment in children up to 16 years during at least one year. The main outcome was caries increment measured by the change in decayed, missing and filled tooth surfaces (D(M)FS). DATA COLLECTION AND ANALYSIS: Inclusion decisions, quality assessment and data extraction were duplicated in a random sample of one third of studies, and consensus achieved by discussion or a third party. Study authors were contacted for missing data. The primary outcome measure was the prevented fraction (PF), that is the caries increment in the treatment group expressed as a percentage of the control group. Random effects meta-analyses were performed where data could be pooled. Potential sources of heterogeneity were examined in random effects meta-regression analyses. MAIN RESULTS: Twenty-five studies were included, involving 7747 children. For the 23 that contributed data for meta analysis, the D(M)FS pooled prevented fraction estimate was 28% (95% CI, 19% to 37%; p<0.0001). There was clear heterogeneity, confirmed statistically (p<0.0001). The effect of fluoride gel varied according to type of control group used, with D(M)FS PF on average being 19% (95% CI, 5% to 33%; p<0.009) higher in non-placebo controlled trials. A funnel plot of the 23 studies indicated a relationship between prevented fraction and study precision. Only two trials reported on adverse events. REVIEWER'S CONCLUSIONS: There is clear evidence of a caries-inhibiting effect of fluoride gel. The best estimate of the magnitude of this effect, based on the 14 placebo-controlled trials, is a 21% reduction (95% CI, 14 to 28%) in D(M)FS. This corresponds to an NNT of 2 (95% CI, 1 to 3) to avoid 1 D(M)FS in a population with a caries increment of 2.2 D(M)FS/year, or an NNT of 24 (95% CI, 18 to 36) based on an increment of 0.2 D(M)FS/year. There is little information concerning deciduous dentition, on adverse effects or on acceptability of treatment. Future trials should include assessment of potential adverse effects. PMID- 12076447 TI - Antidepressants for people with both schizophrenia and depression. AB - BACKGROUND: Depressive symptoms, often of substantial severity, are found in 50% of newly diagnosed suffers of schizophrenia and 33% of people with chronic schizophrenia who have relapsed. Depression is associated with dysphoria, disability, reduction of motivation to accomplish tasks and the activities of daily living, an increased duration of illness and more frequent relapses. OBJECTIVES: To determine the clinical effects of antidepressant medication for the treatment of depression in people who also suffer with schizophrenia. SEARCH STRATEGY: We undertook electronic searches of the Cochrane Schizophrenia Group's Register (October 2000), ClinPsych (1988-2000), The Cochrane Library (Issue 3, 2000), EMBASE (1980-2000) and MEDLINE (1966-2000). This was supplemented by citation searching, personal contact with authors and pharmaceutical companies. SELECTION CRITERIA: All randomised clinical trials that compared antidepressant medication with placebo for people with schizophrenia or schizoaffective disorder who were also suffering from depression. DATA COLLECTION AND ANALYSIS: Data were independently selected and extracted. For homogeneous dichotomous data the fixed effects risk difference (RD), the 95% confidence intervals (CI) and, where appropriate, the number needed to treat (NNT) were calculated on an intention-to treat basis. For continuous data, reviewers calculated weighted mean differences. Statistical tests for heterogeneity were also undertaken. MAIN RESULTS: Eleven studies met the inclusion criteria. All were small, and randomised fewer than 30 people to each group. Most included people after the most acute phase of psychosis and investigated a wide range of antidepressants. The quality of reporting varied a great deal. For the outcome of 'no important clinical response' antidepressants were significantly better than placebo (n=209, 5 RCTs, summary risk difference fixed effects -0.26, 95% CI -0.39 to -0.13, NNT 4 95% CI 3 to 8). The depression score at the end of the trial, as assessed by the Hamilton Rating Scale (HAM-D), seemed to suggest that using antidepressants was beneficial, but this was only statistically significant when a fixed effects model was used (n=261, 6 RCTs, WMD fixed effects -2.2 95% CI -3.8 to -0.6; WMD random effects -2.1 95% CI -5.04 to 0.84). There was no evidence that antidepressant treatment led to a deterioration of psychotic symptoms in the included trials. Heterogeneous data on 'any adverse effect' are equivocal (n=110, 2 RCTs, RD fixed 0.11 CI -0.03 to 0.25, Chi square 7.5, df=1, p=0.0062). In one small study extrapyramidal adverse effects were reported less often by those allocated to antidepressant (n=52, 1 RCT, RD fixed -0.28 CI -0.5 to -0.04). Only about 10% of people left these studies by 12 weeks. There was no apparent difference between those allocated placebo and those given an antidepressant (n=426, 10 RCTs, RD fixed 0.04 CI -0.02 to 0.1). REVIEWER'S CONCLUSIONS: Overall, the literature was of poor quality, and only a small number of trials made useful contributions. Though our results provide some evidence to indicate that antidepressants may be beneficial for people with depression and schizophrenia, the results, at best, are likely to overestimate the treatment effect, and, at worst, could merely reflect selective reporting of statistically significant results and publication bias. At present, there is no convincing evidence to support or refute the use of antidepressants in treating depression in people with schizophrenia. We need further well-designed, conducted and reported research to determine the best approach towards treating depression in people with schizophrenia. PMID- 12076448 TI - Tricyclic drugs for depression in children and adolescents. AB - BACKGROUND: There is a need to identify effective and safe treatments for depression in children and adolescents. While tricyclic drugs are effective in treating depression in adults, individual studies involving children and adolescents have been equivocal. OBJECTIVES: To assess the effects of oral tricyclic antidepressants compared to placebo in the treatment of child and adolescent depression. SEARCH STRATEGY: We searched MEDLINE (1966-1997), EMBASE, Excerpta Medica (June 1974-1997), the Cochrane Collaboration Depression, Anxiety and Neurosis Group trials register (most recent search 25/1/2000) and bibliographies of previously published reviews and papers describing original research were cross-checked. Current Contents was screened for recent publications. We contacted authors of relevant abstracts in conference proceedings of the American Academy of Child and Adolescent Psychiatry, and we hand searched the Journal of the American Academy of Child and Adolescent Psychiatry (1978-1999). SELECTION CRITERIA: Randomised controlled trials comparing the efficacy of orally administered tricyclic medication with placebo in depressed people aged 6-18 years. DATA COLLECTION AND ANALYSIS: Most studies reported multiple outcome measures including depression scales and clinical global impression scales. For each study the best available depression measure was taken as the index measure of depression outcome. Predetermined criteria were established to assist in the ranking of measures. Where authors reported categorical outcomes we calculated individual and pooled odds ratios for the odds of improvement in treated compared with control subjects. For continuous outcomes pooled effect sizes were calculated as the number of standard deviations by which the change in depression scores for the treatment group exceeded those for the control groups. MAIN RESULTS: Thirteen trials (involving 506 participants) were included. No overall improvement with treatment compared to placebo was seen for children or adolescents (odds ratio = 0.84, 95% confidence interval 0.56 to 1.25). A statistically significant but small benefit of treatment over placebo was seen in reducing symptoms (effect size (standardised mean difference) = 0.31, 95% confidence interval -0.62 to -0.01). Subgroup analyses suggest a larger benefit among adolescents (effect size = -0.47, 95% confidence interval -0.92 to 0.02), and no benefit among children (effect size = 0.15, 95% confidence interval -0.34 to 0.64). Treatment with a tricyclic antidepressant caused more vertigo (odds ratio = 4.38, 95% confidence interval 2.33 to 8.25), orthostatic hypotension (odds ratio = 6.78, 95% confidence interval 2.06 to 22.26), tremor (odds ratio 6.29, 95% confidence interval 1.78 to 22.17) and dry mouth (odds ratio = 5.17, 95% confidence interval 2.68 to 29.99) than did placebo, but no statistically significant difference was found for other possible adverse effects. REVIEWER'S CONCLUSIONS: Data suggest tricyclic antidepressants are not useful in treating depression in pre pubertal children. There is marginal evidence to support the use of tricyclic antidepressants in the treatment of depression in adolescents, although the magnitude of effect is likely to be moderate at best. PMID- 12076449 TI - Physical training for cystic fibrosis. AB - BACKGROUND: Physical training may form an important part of the care package for patients with cystic fibrosis. OBJECTIVES: To determine whether a prescribed regime of physical training produces improvement or prevents deterioration in physiological and clinical outcomes in cystic fibrosis compared to no training. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group specialist trials register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. Date of the most recent search: August 2001. SELECTION CRITERIA: All randomised controlled clinical trials in which a prescribed regime of physical training is compared to no physical training in patients with cystic fibrosis. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion, assessed methodological quality and extracted data. MAIN RESULTS: Of the sixteen trials identified six trials, which included 184 patients, met the inclusion criteria. Exercise tolerance in the short term, during an acute admission was improved in patients who received physical training. In a three year study there was no improvement in exercise tolerance at the end of the study period. Over three years there was a significantly greater mean annual rate of decline in FVC in the control group compared to the exercise group WMD 2.17 (95%CI 0.47, 3.87). Changes in other lung function parameters showed a similar trend, but these were not statistically significant. REVIEWER'S CONCLUSIONS: Conclusions about the efficacy of physical training in cystic fibrosis are limited by the small size, short duration and incomplete reporting of most of the trials included in this review. Physical training is already part of the care package offered to most patients with cystic fibrosis and there is no evidence to actively discourage this. Further research is needed to assess comprehensively the net benefit of the addition of physical training to the care of cystic fibrosis patients. PMID- 12076450 TI - "Scared Straight" and other juvenile awareness programs for preventing juvenile delinquency. AB - BACKGROUND: 'Scared Straight' and other programmes involve organised visits to prison by juvenile delinquents or children at risk for criminal behavior. programmes are designed to deter participants from future offending through first hand observation of prison life and interaction with adult inmates. These programmes remain in use world-wide despite studies and reviews questioning their effectiveness. OBJECTIVES: To assess the effects of programmes comprising organised visits to prisons by juvenile delinquents (officially adjudicated or convicted by a juvenile court) or pre-delinquents (children in trouble but not officially adjudicated as delinquents), aimed at deterring them from criminal activity. SEARCH STRATEGY: Handsearching by the first author in identifying randomised field trials 1945-1993 relevant to criminology was augmented by structured searches of 16 electronic data bases, including the Campbell SPECTR database of trials and the Cochrane CCTR. Experts in the field were consulted and relevant citations were followed up. SELECTION CRITERIA: Studies that tested the effects of any program involving the organised visits of juvenile delinquents or children at-risk for delinquency to penal institutions were included. Studies that included overlapping samples of juvenile and young adults (e.g. ages 14-20) were included. We only considered studies that randomly or quasi-randomly (i.e. alternation) assigned participants to conditions. Each study had to have a no treatment control condition with at least one outcome measure of "post-visit" criminal behavior. DATA COLLECTION AND ANALYSIS: We report narratively on the nine eligible trials. We conducted one meta-analysis of post-intervention offending rates using official data. Information from other sources (e.g. self report) was either missing from some studies or critical information was omitted (e.g. standard deviations). We examined the immediate post-treatment effects (i.e. "first-effects") by computing Odds Ratios (OR) for data on proportions of each group re-offending, and assumed both fixed and random effects models in our analyses. MAIN RESULTS: The analysis show the intervention to be more harmful than doing nothing. The program effect, whether assuming a fixed or random effects model, was nearly identical and negative in direction, regardless of the meta-analytic strategy. REVIEWER'S CONCLUSIONS: We conclude that programmes like 'Scared Straight' are likely to have a harmful effect and increase delinquency relative to doing nothing at all to the same youths. Given these results, agencies that permit such programmes must rigorously evaluate them not only to ensure that they are doing what they purport to do (prevent crime) - but at the very least they do not cause more harm than good. PMID- 12076451 TI - Propylthiouracil for alcoholic liver disease. AB - BACKGROUND: Alcohol is the most common cause of liver disease in the Western world today. Randomised clinical trials have addressed the question whether propylthiouracil has any efficacy in patients with alcoholic liver disease. OBJECTIVES: The objectives were to assess the efficacy of propylthiouracil on mortality, clinical symptoms and complications, liver biochemistry, and liver histology in patients with alcoholic liver disease. Adverse events were also analysed. SEARCH STRATEGY: The Cochrane Hepato-Biliary Group Controlled Trials Register (searched July 2001), The Cochrane Controlled Trials Register (Cochrane Library Issue 3, 2001), MEDLINE (January 1966 to July 2001), EMBASE (January 1985 to July 2001) were searched. These electronic searches were combined with full text searches. Manufacturers and researchers in the field were also contacted. SELECTION CRITERIA: Randomised clinical trials studying patients with alcoholic steatosis, alcoholic fibrosis, alcoholic hepatitis, and/or alcoholic cirrhosis were included. Interventions encompassed propylthiouracil at any dose versus placebo or no intervention. The trials could be double-blind, single-blind, or unblinded. The trials could be unpublished or published as an article, an abstract, or a letter and no language limitations were applied. DATA COLLECTION AND ANALYSIS: All analyses were performed according to the intention-to-treat method. The statistical package (RevMan and MetaView) provided by the Cochrane Collaboration was used. The methodological quality of the randomised clinical trials was evaluated by components of quality and the Jadad-scale. MAIN RESULTS: Combining the results of six randomised clinical trials including 710 patients demonstrated no significant effects of propylthiouracil versus placebo on mortality (Peto odds ratio (OR) 0.91, 95% confidence interval (CI) 0.59 to 1.40), liver related mortality (OR 0.78, 95% CI 0.45 to 1.33), complications of the liver disease (OR 1.14, 95% CI 0.58 to 2.24), or liver histology. Propylthiouracil was associated with a non significant trend towards an increased risk of non-serious adverse events (OR 1.49, 95% CI 0.74 to 2.99) and with the seldom occurrence of serious adverse events (leukopenia). REVIEWER'S CONCLUSIONS: This systematic review could not demonstrate any significant efficacy of propylthiouracil on any clinically important outcomes (mortality, liver related mortality, liver complications, and liver histology) of patients with alcoholic liver disease and propylthiouracil was associated with adverse events. Accordingly, there is no evidence for using propylthiouracil for alcoholic liver disease outside randomised clinical trials. PMID- 12076452 TI - Second-line chemotherapy for non-small cell lung cancer. AB - BACKGROUND: The role of second-line chemotherapy for the treatment of patients with non-small cell lung cancer (NSCLC) who have relapsed or failed to respond to first-line treatment was unclear. OBJECTIVES: To determine the effectiveness of any second-line chemotherapy in patients with NSCLC. SEARCH STRATEGY: Medline (1966-July 2001), Embase (1974-July 2001), Cancerlit (1993-July) and the Cochrane Controlled Trials Register (CENTRAL, issue 2 2001) were searched. In addition a handsearch was performed and experts in the field contacted to identify any further studies that had not been found by the electronic searches. SELECTION CRITERIA: Randomised controlled clinical trials in which any second-line chemotherapy was compared with placebo or best supportive care in patients with NSCLC who had previously failed to any previous chemotherapy regimen. DATA COLLECTION AND ANALYSIS: Data were extracted by 2 independent reviewers and revised by a third author. MAIN RESULTS: Only one study was included. This study included a total of 204 patients who were randomised to receive either doxetaxel or best supportive care. Following an unacceptably high toxic death rate the dose of doxetaxel was reduced from 100 mg/m(2) to 75 mg/m(2). Docetaxel gave an extra 2.4 months survival - an average of 7.0 months vs 4.6 months on best supportive care. At 1 year after diagnosis 29% of doxetaxel treated patients were alive compared with 19% of the best supportive care group. REVIEWER'S CONCLUSIONS: Definitive recommendations cannot be made since evidence is only available from one randomised controlled trial which, though of reasonable quality had a number of limitations. There is currently no evidence to support second-line treatment of patients with poor performance status. Larger, well-designed controlled trials are needed to further evaluate whether the benefits of second-line chemotherapy to patients with non-small cell lung cancer outweigh its risks and costs. PMID- 12076453 TI - Embryo freezing for preventing Ovarian Hyperstimulation Syndrome. AB - BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is an iatrogenic potentially life threatening condition resulting from an excessive ovarian stimulation. Its reported incidence varies from 1% to 10% of in vitro fertilization (IVF) cycles. The factors leading to this syndrome have not been completely explained. It seems likely that the release of vasoactive substances, secreted by the ovaries under human chorionic gonadotropin (hCG) stimulation plays a key role in triggering this syndrome. The hallmark of this condition, is a massive shift of fluid from the intra-vascular compartment to the third space resulting in profound intra vascular depletion and hemoconcentration. OBJECTIVES: To evaluate (i) the effectiveness of cryopreservation (embryo freezing) for the prevention of OHSS when compared with human intra-venous albumin infusion and (ii) the effectiveness of the elective cryopreservation (embryo freezing ) of all embryos for the prevention of OHSS when compared with fresh embryo transfer. SEARCH STRATEGY: Publications in the literature that describe or may describe randomised controlled trials of both human intra-venous albumin and freezing of all embryos in the management of OHSS as consequence of the superovulation in assisted reproduction techniques (ART) cycles were searched. The Cochrane Menstrual Disorders and Subfertility Review Group specialised register of controlled trials was searched. In addition, MEDLINE (PUBMED 1985 to 2001), EMBASE (1985 to 2001), CINHAL (1985 to 2001) and the National Research Register were searched SELECTION CRITERIA: Randomised controlled trials (RCTs) in which either human intra-venous albumin or cryopreservation of all embryos were used as a therapeutic approach to OHSS were included. The participants were women of reproductive age who were down regulated by gonadotrophin-releasing hormone-analogue (GnRH-a), undergoing superovulation in in vitro fertilization/intra-cytoplasmic sperm injection (IVF/ICSI) cycles. DATA COLLECTION AND ANALYSIS: Two reviewers, Mr N.N. Amso (NNA) and Dr A. D'Angelo (ADA) scanned the titles and the abstracts of the reports identified by electronic searching in order to find relevant papers. One reviewer (ADA) obtained copies of the full text articles and made copies for the other reviewer (NNA) in which details of authors, institution, results and discussion were removed in order to assess their eligibility for inclusion. Then, both reviewers extracted data independently using forms designed according to Cochrane guidelines. Disagreements were resolved by discussion. Additional information on the trial methodology or data were requested by writing to the corresponding authors directly. The interventions compared were cryopreservation (embryo freezing) versus intra-venous human albumin administration and elective cryopreservation of all embryos versus fresh embryo transfer. The primary outcomes were: incidence of moderate and severe OHSS versus nil/mild OHSS, clinical pregnancies/woman. The secondary outcomes were: number of oocytes retrieved, number of oocytes fertilized, number of embryos transferred, number of embryos frozen, multiple pregnancy rate, live birth rate, number of women admitted to the hospital as inpatient or outpatient and time to the next menstrual period (resolution time). Statistical analysis was performed in accordance with the Cochrane Menstrual Disorders and Subfertility Group guidelines. MAIN RESULTS: Seventeen studies were identified, two of which met our inclusion criteria. One study was included where cryopreservation (embryo freezing) was compared with intra-venous human albumin administration (Shaker 1996) and one study was included where elective cryopreservation of all embryos was compared with fresh embryo transfer (Ferraretti 1999). When cryopreservation was compared with intra-venous human albumin administration no difference was found in all the outcomes examined between the two groups. When elective cryopreservation of all embryos was compared with fresh embryo transfer no difference was found in all the outcomes examined between the two groups. REVIEWER'S CONCLUSIONS: This review has showed that there is insufficient evidence to support routine cryopreservation and insufficient evidence for the relative merits of intra-venous albumin versus cryopreservation. PMID- 12076454 TI - Thermotherapy for treating rheumatoid arthritis. AB - BACKGROUND: Thermotherapy is often used as adjunct in the treatment of rheumatoid arthritis (RA) by rehabilitation specialists. OBJECTIVES: To evaluate the effectiveness of different thermotherapy applications on objective and subjective measures of disease activity in patients with RA. SEARCH STRATEGY: We searched Medline, EMBASE, Pedro, Current Contents, Sports Discus and CINAHL up to and including September 2001. The Cochrane Field of Rehabilitation and related therapies and the Cochrane Musculoskeletal Review Group were also contacted for a search of their specialized registers. Hand searching was conducted on all retrieved articles for additional articles. SELECTION CRITERIA: Comparative controlled studies, such as randomized controlled trials, controlled clinical trials, cohort studies or case/control studies, of thermotherapy compared to control or active interventions in patients with RA were eligible. No language restrictions were applied. Abstracts were accepted. DATA COLLECTION AND ANALYSIS: Two independent reviewers identified potential articles from the literature search (VR, LB). These reviewers extracted data using pre-defined extraction forms. Consensus was reached on all data extraction. Quality was assessed by two reviewers using a 5 point scale that measured the quality of randomization, double-blinding and description of withdrawals. MAIN RESULTS: Seven studies (n=328 subjects) met the inclusion criteria. The results of this systematic review of thermotherapy for RA found that there was no significant effect of hot and ice packs applications (Ivey 1994), cryotherapy (Rembe 1970) and faradic baths (Hawkes 1986) on objective measures of disease activity including joint swelling, pain, medication intake, range of motion (ROM), grip strength, hand function compared to a control (no treatment) or active therapy. There is no significant difference between wax and therapeutic ultrasound as well as between wax and faradic bath combined to ultrasound for all the outcomes measured after 1, 2 or 3 week(s) of treatment (Hawkes 1986). There was no difference in patient preference for all types of thermotherapy. No harmful effects of thermotherapy were reported. REVIEWER'S CONCLUSIONS: Superficial moist heat and cryotherapy can be used as a palliative therapy. Paraffin wax baths combined with exercises can be recommended for beneficial short term effects for arthritic hands. These conclusions are limited by methodological considerations such as the poor quality of trials. PMID- 12076455 TI - Psychoeducation for schizophrenia. AB - BACKGROUND: Schizophrenia can be a severe and chronic illness characterised by lack of insight and poor compliance with treatment. Psychoeducational approaches have been developed to increase patients' knowledge of, and insight into, their illness and its treatment. It is supposed that this increased knowledge and insight will enable people with schizophrenia to cope in a more effective way with their illness, thereby improving prognosis. OBJECTIVES: To assess the effects of psychoeducational interventions compared to the standard levels of knowledge provision. SEARCH STRATEGY: Electronic searches of CINAHL (1982-1999), The Cochrane Library CENTRAL (Issue 1, 1999), The Cochrane Schizophrenia Group's Register (May 2001), EMBASE (1980-1999), MEDLINE (1966-1999), PsycLit (1974 1999), and Sociofile (1974-1999) were undertaken. These were supplemented by cross-reference searching and personal contact with authors of all included studies. SELECTION CRITERIA: All relevant randomised controlled trials focusing on psychoeducation for schizophrenia and/or related serious mental illnesses involving individuals or groups. Quasi-randomised trials were excluded. DATA COLLECTION AND ANALYSIS: Data were extracted independently from included papers by at least two reviewers. Authors of trials were contacted for additional and missing data. Relative risks (RR) and 95% confidence intervals (CI) of homogeneous dichotomous data were calculated. A random effects model was used for heterogeneous dichotomous data. Where possible the numbers needed to treat (NNT) were also calculated. Weighted or standardised means were calculated for continuous data. MAIN RESULTS: Ten studies are included in this review. All studies of group education included family members. Compliance with medication was significantly improved in a single study using brief group intervention (at one year) but other studies produced equivocal or skewed data. Any kind of psychoeducational intervention significantly decreased relapse or readmission rates at nine to 18 months follow-up compared with standard care (RR 0.8 CI 0.7 0.9 NNT 9 CI 6-22). Several of the secondary outcomes (knowledge gain, mental state, global level of functioning, expressed emotion in family members) were measured using scales that are difficult to interpret. Generally, however, findings were consistent with the possibility that psychoeducation has a positive effect on a persons' well being. No impact was found on insight, medication related attitudes or on overall satisfaction with services of patients or relatives but these findings rested on very few studies. Health economic outcome was only measured in one study and data were skewed. It was not possible to analyse whether different duration or formats of psychoeducation influenced effectiveness. REVIEWER'S CONCLUSIONS: Evidence from trials suggests that psychoeducational approaches are useful as a part of the treatment programme for people with schizophrenia and related illness. The fact that the interventions are brief and inexpensive should make them attractive to managers and policy makers. More well-designed, conducted and reported randomised studies investigating the efficacy of psychoeducation are needed. PMID- 12076456 TI - Haloperidol for agitation in dementia. AB - BACKGROUND: Agitation occurs in up to 70% of demented patients. Haloperidol has been used for decades to control agitation in dementia, but its effectiveness remains unclear. Previous meta-analyses examined only English language publications or compared haloperidol with other drugs rather than with placebo. To study the effectiveness of haloperidol a more widely based review was performed. OBJECTIVES: To determine whether evidence supported the use of haloperidol in agitated dementia. SEARCH STRATEGY: The CDCIG Specialized Register which contains references from medical databases (MEDLINE, EMBASE, PsycInfo and CINAHL) as well as from many trials databases was searched on 26 July 2000 to identify reports of randomised controlled trials on haloperidol treatment of agitation in dementia. SELECTION CRITERIA: Randomized, placebo-controlled trials, with concealed allocation, where subjects' dementia and agitation were assessed. DATA COLLECTION AND ANALYSIS: 1. Two reviewers extracted data from included trials 2. Data were pooled where possible, and analysed using appropriate statistical methods 3. Odds ratios of average differences were calculated 4. Only 'intention to treat' data were included 5. Analysis included haloperidol treated patients, compared with placebo MAIN RESULTS: The five included trials led to the following results: 1. There was no significant improvement in agitation among haloperidol treated patients, compared with controls. 2. Aggression decreased among patients with agitated dementia treated with haloperidol; other aspects of agitation were not affected significantly in treated patients, compared with controls. 3. Although two studies showed increased dropouts due to adverse effects among haloperidol patients, there was no significant difference in dropout rates, comparing all haloperidol treated patients with controls. 4. The data were insufficient to examine response to treatment in relation to length of treatment, degree of dementia, age or sex of patients, and cause of dementia. REVIEWER'S CONCLUSIONS: 1. Evidence suggests that haloperidol was useful in the control of aggression, but was associated with increased side effects; there was no evidence to support the routine use of this drug for other manifestations of agitated dementia. 3. Similar dropout rates among haloperidol and placebo treated patients suggested that poorly controlled symptoms, or other factors, may be important in causing treatment discontinuation. 4. Variations in degree of dementia, dosage and length of haloperidol treatment, and in ways of assessing response to treatment suggested caution in the interpretation of reported effects of haloperidol in the management of agitated dementia. 4. The present study confirmed that haloperidol should not be used routinely to treat patients with agitated dementia. Treatment of agitated dementia with haloperidol should be individualized and patients should be monitored for side effects of therapy. PMID- 12076457 TI - Interventions for preventing eating disorders in children and adolescents. AB - BACKGROUND: Eating disorders represent an extremely difficult condition to treat and patients consume an enormous amount of mental health energy and resources. Being young, female, and dieting are some of the few identified risk factors that have been reliably linked to the development of eating disorders, and several prevention eating disorder prevention programs have been developed and trialed with children and adolescents. The purpose of this systematic review is to evaluate the effectiveness of eating disorder prevention programs for children and adolescents both in the general population and those determined to be at risk. OBJECTIVES: 1. To determine if eating disorder prevention programs are effective in promoting healthy eating attitudes and behaviours in children and adolescents; 2. To determine if eating disorder prevention programs are effective in promoting psychological factors that protect children and adolescents from developing eating disorders; 3. To determine if eating disorder prevention programs are effective in promoting satisfactory physical health in children and adolescents; 4. To determine if eating disorder prevention programs have a long term, sustainable, and positive impact on the mental and physical health of children and adolescents; and, 5. To determine the safety of eating disorder prevention programs in terms of possible harmful consequences on the mental or physical health of children and adolescents. SEARCH STRATEGY: Relevant trials are identified through searching the Cochrane Controlled Trial Register (CCTR) and relevant biomedical and social science databases. All terms necessary to detect prevention programs and the participant groups are used. A strategy to locate randomised controlled trials is used. Other sources of information are the bibliographies of systematic and non-systematic reviews and reference lists from articles identified through the search strategy. In order to identify unpublished studies, experts in the field are contacted by letter and/or electronic mail. SELECTION CRITERIA: Randomised controlled trials (RCT) with a major focus on eating disorder prevention programs for children and adolescents, where there is no known DSM-IV diagnosis of an eating disorder, are eligible for inclusion in the review. Trials must include a control group and at least one objective outcome measure (eg. BMI) or a standardised psychological measure used with the intervention and control group, pre- and post-intervention. DATA COLLECTION AND ANALYSIS: A total of 1379 titles have been identified through the search to date. 13 studies were located that reported use of a randomised controlled trial methodology and were critically appraised by two independent reviewers. Five (5) studies were excluded as data were not reported in a useable form or useable data could not be obtained from the trial authors, one dissertation could not be obtained, one study had no "true" no-treatment or usual treatment control group, and one study did not use a pre-test outcome measure. Eight (8) studies met the selection criteria outlined above. MAIN RESULTS: Only one of eight pooled comparisons of two or more studies using similar outcome measures and similar intervention types demonstrated the statistically significant effect of a particular type of eating disorder prevention program for children and adolescents. Combined data from two eating disorder prevention programs based on a media literacy and advocacy approach indicate a reduction in the internalisation or acceptance of societal ideals relating to appearance at a 3- to 6-month follow-up (Kusel, unpublished; Neumark-Sztainer2000) [SMD -0.28, -0.51 to -0.05, 95% CI]. However, there is insufficient evidence to conclude that this approach also demonstrated a significant impact on awareness of societal standards relating to appearance. There is insufficient evidence to support the effect of four programs designed to address eating attitudes and behaviours and other adolescent issues on body weight, eating disorder symptoms, associated eating disorder psychopathology or general psychological and physical well-being in the general sample or those classified as being at high risk for eating disorder (Buddeberg-F 1998; Killen 1993/1996; Santonastaso 1999; Zanetti 1999). Given only one program used a psychoeducation approach to prevent bulimia nervosa (Jerome, unpublished) and only one program adopted a focus on self-esteem (O'Dea 2000), the effect of these approaches could not be evaluated via meta-analyses. In relation to potential harmful effects, there is not sufficient evidence to suggest that harm resulted from any of the prevention programs included in the review. REVIEWER'S CONCLUSIONS: The one significant pooled effect in the current review does not allow for any firm conclusions to be made about the impact of prevention programs for eating disorders in children and adolescents, although none of the pooled comparisons indicated evidence of harm. From a clinical perspective, the development and refinement of prevention programs is complicated by a lack of knowledge about risk factors associated with eating disorders and the need to strike a balance between delivering preventive interventions for eating disorders and considering the potential to cause harm. From a research perspective, the idea of "thresholds" for identifying young people at risk of developing eating disorders has been raised, and denial of concern or denial of illness represents a further issue complicating early identification in relation to eating disorder symptomatology. Longer-term effects of the intervention approaches will need to be monitored across development in order to demonstrate a decline in the incidence of eating disorders and associated risk factors. PMID- 12076458 TI - Antibiotic prophylaxis for cirrhotic patients with gastrointestinal bleeding. AB - BACKGROUND: Bacterial infections are a frequent complication in patients with cirrhosis and gastrointestinal bleeding. Antibiotic prophylaxis seems to decrease the incidence of bacterial infections. Oral antibiotics, active against enteric bacteria, have been most often used as antibiotic prophylaxis in cirrhotic patients with gastrointestinal bleeding. OBJECTIVES: This review aims to evaluate the effects of antibiotic prophylaxis in the prevention of bacterial infections in cirrhotic patients with gastrointestinal bleeding. SEARCH STRATEGY: Electronic searches on The Cochrane Hepato-Biliary Group Controlled Trials Register (May 2001), The Cochrane Library (Issue 2, 2001), EMBASE (1980-2001), and MEDLINE (1966-2001); handsearching the references of all identified studies; contacting the first author of each included trial. SELECTION CRITERIA: Randomised clinical trials comparing different types of antibiotic prophylaxis with placebo, no intervention, or another antibiotic to prevent bacterial infections in cirrhotic patients with gastrointestinal bleeding. DATA COLLECTION AND ANALYSIS: Two reviewers independently appraised the quality of each trial and extracted the data from the included trials. Relative risks (RR) or average differences, with their 95% confidence intervals (CI) were estimated. The reviewers assumed an intention to treat basis for the outcome measures. MAIN RESULTS: Eight trials evaluated the effects of antibiotic prophylaxis compared with placebo or no antibiotic prophylaxis in 864 patients. A significant beneficial effect on decreasing mortality (RR 0.73, 95% CI 0.55 to 0.95) and the incidence of bacterial infections (RR 0.40, 95% CI 0.32 to 0.51) was observed. No serious adverse events were reported. The trials showed no significant heterogeneity. Three additional trials evaluated the effects of antibiotics compared with a different regimen of antibiotics in 503 patients. Data could not be combined as each trial used different interventions. None of the examined antibiotic regimens was superior to the control regimen regarding mortality or the incidence of bacterial infections. REVIEWER'S CONCLUSIONS: Antibiotic prophylaxis for cirrhotic inpatients with gastrointestinal bleeding is efficacious in reducing the number of deaths and bacterial infections, are well tolerated, and should be advocated. PMID- 12076459 TI - Nutrition support for bone marrow transplant patients. AB - BACKGROUND: Bone marrow transplantation involves the administration of toxic chemotherapy and infusion of marrow cells. After treatment, patients can develop a poor appetite, mucositis and gastrointestinal failure, leading to malnutrition. To prevent this, parenteral nutrition (PN) support is the first choice but is associated with an increased risk of infection. Enteral nutrition (EN) is an alternative, as is the addition of substrates e.g. glutamine to enteral and parenteral solutions. However, the relative effectiveness of these treatments is not clear. OBJECTIVES: To determine the efficacy of EN or PN support for patients receiving a bone marrow transplant. SEARCH STRATEGY: Trials were identified by searching the Cochrane Library (Issue 4, 2000 ), MEDLINE (1966-2000), EMBASE (1988-2000) and CINAHL (1982-2000 ). Reference lists of identified trials and conference proceedings were searched for relevant reports. Date of the most recent search: 2000. SELECTION CRITERIA: RCTs that compared one form of nutrition support with another, or control, for bone marrow transplant patients were included. DATA COLLECTION AND ANALYSIS: Thirty five reports were identified, 11 were excluded. Two reviewers extracted data from 24 studies; 16 were allocated to four interventions: oral glutamine versus placebo; PN and glutamine versus standard PN; PN versus IV hydration; PN versus EN. Eight studies were other interventions. Data were collected on participants' characteristics; adverse effects; neutropaenia; % change in body weight; graft versus host disease; and survival. Trialists were contacted for unreported data. MAIN RESULTS: Two studies (82 subjects) found that glutamine mouthwash reduced days of neutropaenia (6.82 days, 95% CI (1.67-11.98) p=0.009) compared with placebo. Three studies (103 subjects) showed that patients receiving PN with glutamine had a reduced hospital stay, 6.62 d (95% CI 3.47, 9.77, P=0.00004) compared with patients receiving standard PN. Two studies (73 subjects) indicated that patients receiving PN plus glutamine had a reduced incidence of positive blood cultures (OR 0.23, 95% CI 0.08-0.65, p=0.006) compared to those receiving standard PN. One study, (25 subjects) showed patients receiving PN had a higher incidence of line infections (odds ratio 21.23, 95% CI 4.15,108.73, P=0.0002) compared to those receiving standard IV fluids. There were no evaluable data to compare PN with EN. REVIEWER'S CONCLUSIONS: Lack of evaluable data means that the relative effectiveness of EN versus PN cannot be evaluated. Further studies and missing data from completed trials need to be retrieved. Studies comparing PN with glutamine versus standard PN suggest that patients leave hospital earlier, and experience a reduced incidence of positive blood cultures, than those receiving standard PN. Patients with gastrointestinal failure should consider PN with the addition of glutamine if enteral feeding is not possible. PMID- 12076460 TI - In-hospital care pathways for stroke. AB - BACKGROUND: Care within a stroke unit reduces death or dependency after stroke. However, studies have found significant variations in clinical practice, access to stroke unit care, organisation of patient care, and clinical outcome. Stroke care pathways have been introduced as a method to promote organised and efficient patient care that is based on best evidence and guidelines. OBJECTIVES: We aimed to assess the effects of care pathways, as compared to standard medical care, among patients with acute stroke who had been admitted to hospital. SEARCH STRATEGY: We searched the Cochrane Stroke Group Specialised Trials Register (last searched in May 2001), the Cochrane Controlled Trials Register (Issue 4, 2000), MEDLINE (1975-2000), EMBASE (1980-2000), CINAHL (1982-2000), the Index to Scientific and Technical Proceedings (ISTP, May 2001), and HealthSTAR (May 2001). We also handsearched the Journal of Managed Care (1997 to 1998), which was later renamed the Journal of Integrated Care (1998 to 2001). Reference lists of articles were searched. SELECTION CRITERIA: We considered randomised controlled trials and non-randomised studies (quasi-randomised trials, comparative studies, controlled and uncontrolled before and after studies, and interrupted time series) that compared care pathway care with standard medical care. DATA COLLECTION AND ANALYSIS: One reviewer selected studies for inclusion and the other independently checked the decisions. Two reviewers independently assessed the methodological quality of the studies. One reviewer extracted the data and the other checked the extracted data. Data from randomised and non-randomised studies were analysed separately. We found significant statistical heterogeneity in the analysis of two outcomes (computed tomography brain scanning and duration of stay). MAIN RESULTS: There were three randomised controlled trials (total of 340 patients) and seven non-randomised studies (total of 1673 patients) that compared care pathway care with standard medical care. We found no difference between care pathway and control groups in terms of death, dependency, or discharge destination. Evidence from mainly non-randomised studies suggests that patients managed using a care pathway may be: a) less likely to suffer a urinary tract infection (OR 0.38, CI 0.18 to 0.79), b) less likely to be readmitted (OR 0.11, CI 0.03 to 0.39), and c) more likely to have a computed tomography brain scan (OR 3.66, CI 1.45 to 9.27) or carotid duplex study (OR 2.45, CI 1.3 to 4.61). Evidence from randomised trials suggests that patient satisfaction and quality of life may be significantly lower in the care pathway group (P=0.02 and P<0.005 respectively). REVIEWER'S CONCLUSIONS: The use of care pathways to manage stroke patients in hospital may be associated with both positive and negative effects on the process of care and clinical outcomes. Since most of the results have been derived from non-randomised studies, they are likely to be influenced by potential biases and confounding factors. There is currently insufficient supporting evidence to justify routine implementation of care pathways for acute stroke management or stroke rehabilitation. PMID- 12076461 TI - Risk of fatal and nonfatal lactic acidosis with metformin use in type 2 diabetes mellitus. AB - BACKGROUND: Metformin is an oral anti-hyperglycemic agent used in the treatment of type 2 diabetes mellitus. The results of the UK Prospective Diabetes Study indicate that metformin treatment is associated with a reduction in total mortality compared to other anti-hyperglycemic treatments. Metformin, however, is thought to increase the risk of lactic acidosis, and is considered to be contraindicated in many chronic hypoxemic conditions that may be associated with lactic acidosis, such as cardiovascular, renal, hepatic and pulmonary disease, and advancing age. OBJECTIVES: To assess the incidence of fatal and nonfatal lactic acidosis with metformin use compared to placebo and other glucose-lowering treatments in patients with type 2 diabetes mellitus. A secondary objective was to evaluate the blood lactate levels for those on metformin treatment compared to placebo or non-metformin therapies. SEARCH STRATEGY: A search was performed of the Cochrane Controlled Trials Register and the Database of Abstracts of Reviews of Effectiveness (up to 4/2000), Medline (up to 11/2000), Embase (up to 11/2000), Oldmedline, and Reactions (up to 5/2000), in order to identify all studies of metformin treatment from 1966 to November 2000. The Cumulated Index Medicus was used to search relevant articles from 1959 to 1965. The search was augmented by scanning references of identified articles, and by contacting principal investigators. Date of latest search: November 2000. SELECTION CRITERIA: Prospective trials in patients with type 2 diabetes that lasted longer than one month were included if they evaluated metformin, alone or in combination with other treatments, compared to placebo or any other glucose-lowering therapy. Observational cohort studies of metformin treatment lasting greater than one month were also included. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials to be included, assessed study quality and extracted data. The incidence of fatal and nonfatal lactic acidosis was recorded as cases per patient-years, for metformin treatment and for placebo or other treatments. The upper limit for the true incidence of cases in the metformin and non-metformin groups were calculated using Poisson statistics. In a second analysis lactate levels were measured as a net change from baseline or as mean treatment values (basal and stimulated by food or exercise) for treatment and comparison groups. The pooled results were recorded as a weighted mean difference (WMD) in mmol/L, using the fixed effects model for continuous data. MAIN RESULTS: Pooled data from 176 comparative trials and cohort studies revealed no cases of fatal or nonfatal lactic acidosis in 35,619 patient-years of metformin use or in 30,002 patients-years in the non-metformin group. Using Poisson statistics with 95% confidence intervals the upper limit for the true incidence of metformin associated lactic acidosis was 8.4 cases per 100,000 patient-years, and the upper limit for the true incidence of lactic acidosis in the non-metformin group was 9 cases per 100,000 patient-years. There was no difference in lactate levels, either as mean treatment levels or as a net change from baseline, for metformin compared to placebo or other non-biguanide therapies. The mean lactate levels were slightly lower for metformin treatment compared to phenformin (WMD -0.75 mmol/L, 95% CI -0.86 to -0.15). REVIEWER'S CONCLUSIONS: There is no evidence from prospective comparative trials or from observational cohort studies that metformin is associated with an increased risk of lactic acidosis, or with increased levels of lactate, compared to other anti-hyperglycemic treatments if prescribed under the study conditions, taking into account contra-indications. PMID- 12076462 TI - Tracheal gas insufflation for the prevention of morbidity and mortality in mechanically ventilated newborn infants. AB - BACKGROUND: Tracheal gas insufflation (TGI) is a technique where a continuous flow of gas is instilled into the lower trachea during conventional mechanical ventilation. TGI can improve carbon dioxide removal with lower ventilation pressures and smaller tidal volumes, potentially decreasing secondary lung injury and chronic lung disease (CLD). OBJECTIVES: To assess whether, in mechanically ventilated neonates, the use of tracheal gas insufflation reduces mortality, CLD and other adverse clinical outcomes without significant side effects. SEARCH STRATEGY: Searches were made of MEDLINE 1966 to December 2001, CINAHL 1982 to December 2001, the Cochrane Controlled Trials Register (Cochrane Library, Issue 4, 2001) and conference and symposia proceedings. SELECTION CRITERIA: Randomised controlled trials (RCT) that include newborn infants who are mechanically ventilated, and compare TGI during conventional mechanical ventilation (CMV) with CMV alone. Primary outcomes - mortality, CLD and neurodevelopmental outcome; secondary outcomes - air leak, intraventricular haemorrhage, periventricular leukomalacia, duration of mechanical ventilation, duration of respiratory support, duration of oxygen therapy, duration of hospital stay, retinopathy of prematurity, immediate adverse effects. DATA COLLECTION AND ANALYSIS: Each reviewer assessed eligibility, trial quality and extracted data separately. Study authors were contacted for additional information if necessary. MAIN RESULTS: Only one small study was found to be eligible. This study found no evidence of effect on mortality, CLD or age at first extubation. The total duration of ventilation was 9.3 days shorter in the TGI group (95% CI from 15.7 to 2.9 days shorter). The age at complete weaning from ventilation was 26 days shorter in the TGI group (95% CI from 46 to 6 days shorter). There was no evidence of effect on the total duration of respiratory support, oxygen therapy or hospital stay. REVIEWER'S CONCLUSIONS: There is evidence from a single RCT that TGI may reduce the duration of mechanical ventilation in preterm infants - although the data from this small study do not give sufficient evidence to support the introduction of TGI into clinical practice. The technical requirements for performing TGI (as performed in the single included study) are great. There is no statistically significant reduction in the total duration of respiratory support or hospital stay. TGI cannot be recommended for general use at this time. PMID- 12076463 TI - Early versus delayed initiation of continuous distending pressure for respiratory distress syndrome in preterm infants. AB - BACKGROUND: The application of a continuous distending pressure (CDP) has been shown to have some benefits in the treatment of pre-term infants with respiratory distress syndrome (RDS). CDP has the potential to reduce lung damage, particularly if applied early before atelectasis has occurred. Early application of CDP may better conserve an infant's own surfactant stores and consequently be more effective than CDP applied later in the course of RDS. OBJECTIVES: In pre term infants with RDS to determine if early compared with delayed initiation of CDP results in lower mortality and reduced need for intermittent positive pressure ventilation. SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Review Group was used. This included searches of the Oxford Database of Perinatal Trials, Cochrane Controlled Trials Register (Issue 1, 2002), MEDLINE (1966-2001), previous reviews including cross references, abstracts, conference and symposia proceedings, expert informants, journal hand searching mainly in the English language. SELECTION CRITERIA: Trials among pre term infants with respiratory distress syndrome spontaneously breathing at trial entry, which used random or quasi-random allocation to either early or delayed CDP. DATA COLLECTION AND ANALYSIS: Standard methods of the Cochrane Collaboration and its Neonatal Review Group were used, including independent assessment of trial quality and extraction of data by two authors. MAIN RESULTS: In six studies on a total of 165 infants, early CDP was associated with a significant reduction in subsequent use of intermittent positive pressure ventilation, typical RR 0.55, RD -0.16, NNT 6, but there was no evidence of effect on overall mortality. There was no evidence of effect on the rates of pneumothorax (five studies) or bronchopulmonary dysplasia (one study). Early CDP resulted in a reduction in duration of oxygen therapy in the single study reporting this outcome. REVIEWER'S CONCLUSIONS: Early application of CDP has a clinical benefit in the treatment of RDS in that it reduces subsequent use of IPPV and thus may be useful in preventing the adverse effects of this treatment. However, many of the trials were done in the 1970s and 1980s and re-evaluation of the strategy of early CDP in the era of antenatal steroid use and early surfactant administration is indicated. PMID- 12076464 TI - Drug treatments for obstructive sleep apnoea. AB - BACKGROUND: The treatment of choice for moderate to severe obstructive sleep apnoea (OSA) is continuous positive airway pressure (CPAP) via a mask during sleep. However this is not tolerated by all patients and its role in mild OSA is not proven. Drug therapy has been proposed as an alternative to CPAP in some patients with mild to moderate sleep apnoea. The mechanisms by which drugs might reduce OSA include; a reduction in the proportion of rapid eye movement (REM) sleep (during which apnoeas tend to be more frequent), an increase in ventilatory drive or an increase in upper airway muscle tone during sleep. OBJECTIVES: To determine the efficacy of drug therapies in the treatment of sleep apnoea. SEARCH STRATEGY: Searches were carried out on the Cochrane Airways Group RCT Register. Additional hand searching was performed as relevant. SELECTION CRITERIA: Double blind, randomised placebo controlled trials were included, involving patients with confirmed obstructive sleep apnoea. Trials were excluded if continuous positive airways pressure, mandibular devices or oxygen therapy were used. No restriction was placed upon publication language or trial duration. DATA COLLECTION AND ANALYSIS: A total of 51 references were identified by electronic searches. 42 studies were retrieved for selection and 9 trials were included in the review. The results for 91 patients were available. No response for further information was forthcoming from the study authors. Results were expressed as (WMD) and 95% Confidence Intervals (95% CI) MAIN RESULTS: Only acetazolamide reduced the Hypopnoea Index (1 crossover trial of 9 patients, Weighted Mean Difference -24; 95%Confidence Intervals (95% CI): -4, -44). However there was no symptomatic response and the drug was poorly tolerated. Protriptyline led to a symptomatic improvement (improved vs not improved) in two out of three crossover trials (13 patients, Peto Odds Ratio 29.2; 95%CI 2.8, 301.1) but there was no change in the apnoea frequency. No beneficial effects were found for medroxy progesterone, clonidine, buspirone, aminophylline, theophylline or sabeluzole. REVIEWER'S CONCLUSIONS: The data available do not support the use of drugs as a therapy for OSA. Although the studies examined had limitations there was little to justify further trials of these particular drugs. PMID- 12076465 TI - Rehabilitation after lumbar disc surgery. AB - BACKGROUND: Although several rehabilitation programs, physical fitness programs or protocols regarding instruction for patients to return to work after lumbar disc surgery have been suggested, little is known about the efficacy and effectiveness of these treatments. There are still persistent fears of causing re injury, re-herniation, or instability. OBJECTIVES: The objective of this systematic review was to evaluate the effectiveness of active treatments that are used in the rehabilitation after first-time lumbar disc surgery. SEARCH STRATEGY: We searched the MEDLINE, EMBASE and Psyclit databases up to April 2000 and the Cochrane Controlled Trials Register 2001, Issue 3. SELECTION CRITERIA: Both randomized and non-randomized controlled trials on any type of active rehabilitation program after first-time disc surgery were included. DATA COLLECTION AND ANALYSIS: Two independent reviewers performed the inclusion of studies and two other reviewers independently performed the methodological quality assessment. A rating system that consists of four levels of scientific evidence summarizes the results. MAIN RESULTS: Thirteen studies were included, six of which were of high quality. There is no strong evidence for the effectiveness for any treatment starting immediately post-surgery, mainly because of lack of (good quality) studies. For treatments that start four to six weeks post-surgery there is strong evidence (level 1) that intensive exercise programs are more effective on functional status and faster return to work (short-term follow-up) as compared to mild exercise programs and there is strong evidence (level 1) that on long term follow up there is no difference between intensive exercise programs and mild exercise programs with regard to overall improvement. For all other primary outcome measures for the comparison between intensive and mild exercise programs there is conflicting evidence (level 3) with regard to long-term follow-up. Furthermore, there is no strong evidence for the effectiveness of supervised training as compared to home exercises. There was also no strong evidence for the effectiveness of multidisciplinary rehabilitation as compared to usual care. There is limited evidence (level 3) that treatments in working populations that aim at return to work are more effective than usual care with regard to return to work. Also, there is limited evidence (level 3) that low tech and high-tech exercises, started more than 12 months post-surgery are more effective in improving low back functional status as compared to physical agents, joint manipulations or no treatment. Finally, there is no strong evidence for the effectiveness of any specific intervention when added to an exercise program, regardless of whether exercise programs start immediately post-surgery or later. None of the investigated treatments seem harmful with regard to re-herniation or re-operation. REVIEWER'S CONCLUSIONS: There is no evidence that patients need to have their activities restricted after first time lumbar disc surgery. There is strong evidence for intensive exercise programs (at least if started about 4-6 weeks post-operative) on short term for functional status and faster return to work and there is no evidence they increase the re-operation rate. It is unclear what the exact content of post-surgery rehabilitation should be. Moreover, there are no studies that investigated whether active rehabilitation programs should start immediately post-surgery or possibly four to six weeks later. PMID- 12076466 TI - Dietary interventions for recurrent abdominal pain (RAP) in childhood. AB - BACKGROUND: Between 4% and 25% of school-age children complain of recurrent abdominal pain (RAP) of sufficient severity to interfere with daily activities. It is unclear whether the diagnosis of RAP includes children with different aetiologies for their pain. For the majority of such children, no organic cause for their pain can be found on physical examination or investigation. Although most children are likely managed by reassurance and simple measures, a large range of interventions have been recommended. OBJECTIVES: To determine the effectiveness of dietary interventions for recurrent abdominal pain in school-age children. SEARCH STRATEGY: The Cochrane Library (CENTRAL), MEDLINE, EMBASE, CINAHL, ERIC, PsycINFO, LILACS and JICST were searched using a strategy combining (Recurrent OR synonyms) AND (Abdomen OR synonyms) AND (Pain OR synonyms). Where appropriate, search filters were employed. In addition, researchers working in this area were asked to identify relevant studies. SELECTION CRITERIA: Any study in which the majority of participants were school-age children fulfilling standard criteria for RAP, and who were allocated by random or quasi-random methods to any dietary treatment compared with a placebo or no treatment, were selected. DATA COLLECTION AND ANALYSIS: References identified by the searches were screened against the inclusion criteria by two independent reviewers. MAIN RESULTS: Four trials were included in the review overall. Two comparing fibre supplements with placebo (Christensen 1982, Feldman 1985) were included, with data from one study reported in two papers (Christensen 1982, Christensen 1986). A total of 92 school-aged children participated in the two trials. The pooled odds ratio for improvement in the frequency of abdominal pain was 1.16 (95% CI 0.45-2.87). Two trials (Lebenthal 1981, Dearlove 1983) comparing lactose containing with lactose-free diets were included. A total of 90 school-aged children participated in these two trials; however, neither trial reported data in a form which could be used in the meta-analysis and the former trial had a loss to follow-up of 45%. We are continuing to try to contact the authors of the Lebenthal 1981 trial for more data. The authors of Dearlove 1983 reported that raw data from their trial is not available. REVIEWER'S CONCLUSIONS: There is a lack of high quality evidence on the effectiveness or otherwise of dietary interventions. This review suggests that fibre supplements are not effective in the management of RAP, although the trials are small and of variable quality. The trials from lactose-restricting diets are as yet wholly inclusive. There is a need for well-designed trials (which include compliance measures) of all recommended dietary interventions for children with RAP. PMID- 12076467 TI - Beta-lactam versus beta-lactam-aminoglycoside combination therapy in cancer patients with neutropaenia. AB - BACKGROUND: Chemotherapy treated cancer patients are prone to neutropaenia and life-threatening infections. Early, empirical antibiotic treatment is therefore administered routinely to febrile neutropaenic patients. Currently, either beta lactam-aminoglycoside combination treatment or beta-lactam monotherapy are recommended. OBJECTIVES: We compared beta-lactam monotherapy versus beta-lactam aminoglycoside combination therapy for cancer patients with fever and neutroepaenia. SEARCH STRATEGY: Cochrane Library (Issue 4,2001), the Cochrane Cancer Network Register of trials (July 2000), EMBASE (January 1980-2000), MEDLINE (1966-8/2001), and ICAAC conference proceedings (1995 onwards). We scanned references of all included studies, pertinent reviews, and contacted the first author of each included trial and the pharmaceutical companies. SELECTION CRITERIA: Randomised controlled trials comparing any beta-lactam antibiotic monotherapy to any combination of a beta-lactam and an aminoglycoside antibiotic, for the initial, empirical treatment of febrile neutropaenic cancer patients. DATA COLLECTION AND ANALYSIS: Data concerning mortality, treatment failure (including treatment modifications), superinfections, adverse effects and study quality measures were extracted independently by two reviewers. Relative risks with their 95% confidence intervals (CI) were estimated. Outcomes were extracted by intention-to-treat analysis whenever possible. MAIN RESULTS: Forty-six trials and 7642 patients were included. All cause mortality was the primary outcome assessed. For all mortality comparisons, no significant difference between monotherapy and combination therapy was seen, relative risk 0.85 (95% CI 0.72 1.02) for all studies combined. Treatment failure was the outcome reported in all included trials. No significant difference between study groups was shown for studies comparing the same beta-lactam, relative risk 1.12 (95% CI 0.96-1.29). A significant advantage to monotherapy was observed for studies comparing different beta-lactams, relative risk 0.86 (95% CI 0.80-0.93). Bacterial and fungal superinfections developed with similar frequencies in the monotherapy and combination treatment groups. Adverse events were significantly more common in the combination treatment group, relative risk 0.83, (95% CI 0.72-0.97). These included events associated with significant morbidity, primarily renal toxicity. Results were consistent for subgroup and sensitivity analyses. REVIEWER'S CONCLUSIONS: We have shown an advantage to broad-spectrum beta-lactam monotherapy over beta-lactam-aminoglycoside combination therapy for febrile neutropaenia. This advantage comprises of 1) a similar, if not better, survival, 2) a significantly lower treatment failure rate, 3) comparable probability for secondary infections and, 4) most importantly, a lower rate of adverse events associated with significant morbidity. Monotherapy can be regarded, therefore, as the standard of care for febrile neutropaenic patients. PMID- 12076468 TI - Mediastinal chest drain clearance for cardiac surgery. AB - BACKGROUND: Cardiac tamponade may occur following cardiac surgery as a result of blood or fluid collecting in the pericardial space compressing the heart and reducing cardiac output. Mediastinal chest drains (including pericardial drains) are inserted as standard post-operative practice following cardiac surgery to assist the clearance of blood from the pericardial space and to prevent cardiac tamponade. Manipulation techniques including milking, stripping, fanfolding and tapping may be applied to the tubes to keep them from blocking. Evidence is required as to the safest and most effective means of preventing chest tube blockage and preventing cardiac tamponade. OBJECTIVES: To compare different methods of chest drain clearance (i.e. varying levels of suction or suction in combination with milking, stripping, fanfolding and tapping of chest drains) in preventing cardiac tamponade in patients following cardiac surgery. SEARCH STRATEGY: We searched the Cochrane Heart Group specialised register, the Cochrane Controlled Trials Register (CCTR) (Issue 1, 2001) The Cochrane Database of Systematic Reviews (CDSR), the Database of Abstracts of Reviews of Effectiveness (DARE), Issue 1, 2001, MEDLINE (1966 to May Week 1, 2001), EMBASE (1980 to 2001 Week 35), CINAHL (1982 to March 2001), the Clinical Trials site of the NIH, (USA) (10.09.01) and reference lists of articles. SELECTION CRITERIA: Randomised, quasi randomised or systematically allocated clinical trials of chest tube manipulation techniques in adults and children with mediastinal chest drains following cardiac surgery were included. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. Study authors were contacted for additional information where required. Adverse effects information was collected from the trials. MAIN RESULTS: Three studies with a total of 471 participants were included. There was no data, however, which could be included in a meta analysis. This was due to inadequate data provision by two of the studies and where adequate data were provided there were no common interventions or outcomes to pool. On the basis of single studies there was no difference between groups on incidence of chest tube blockage, heart rate, cardiac tamponade or incidence of surgical re-entry. REVIEWER'S CONCLUSIONS: There are insufficient studies which compare differing methods of chest drain clearance to support or refute the relative efficacy of the various techniques in preventing cardiac tamponade. Nor can the need to manipulate chest drains be supported or refuted by results from RCT's. PMID- 12076469 TI - Early surfactant administration with brief ventilation vs selective surfactant and continued mechanical ventilation for preterm infants with or at risk for RDS. AB - BACKGROUND: Both early and prophylactic surfactant replacement therapy compared with later selective surfactant administration reduces mortality and pulmonary complications in ventilated infants with respiratory distress syndrome (RDS). Continuous distending pressure (CDP) has also been shown to improve clinical outcomes in preterm infants with RDS. OBJECTIVES: To compare two treatment strategies in preterm infants with, or at risk for, RDS: early surfactant administration with brief mechanical ventilation (less than 1 hour) followed by extubation, vs later, selective surfactant administration, continued mechanical ventilation and extubation from low respiratory support. Two populations of infants receiving early surfactant were considered: spontaneously breathing infants with signs of RDS (surfactant administration during evolution of RDS prior to requiring intubation for respiratory failure) and infants at high risk for RDS (prophylactic surfactant administration within 15 minutes after birth). SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal trials, MEDLINE (1966-December 2001), CINAHL (1982-December 2001), EMBASE (1980-December 2001), Cochrane Controlled Trials Register (The Cochrane Library, Issue 1, 2002), Pediatric Research (1990-2001), abstracts, expert informants and hand searching. No language restrictions were applied. SELECTION CRITERIA: Randomized or quasi randomized controlled clinical trials comparing early surfactant administration with planned brief mechanical ventilation (less than one hour) followed by extubation, vs selective surfactant administration, continued mechanical ventilation and extubation from low respiratory support. DATA COLLECTION AND ANALYSIS: Data were sought regarding effects on incidence of mechanical ventilation (ventilation continued or initiated beyond one hour after surfactant administration), incidence of bronchopulmonary dysplasia (BPD, need for oxygen at 28 days of age), incidence of chronic lung disease (CLD, need for oxygen at 36 weeks' post-conceptional age), mortality (neonatal mortality < 28 days and mortality prior to hospital discharge), duration of mechanical ventilation, duration of hospitalization, time in oxygen, duration of respiratory support (including CPAP and nasal cannula), number of patients receiving surfactant, number of surfactant doses administered per patient, incidence of air leak syndromes (pulmonary interstitial emphysema, pneumothorax), incidence of pulmonary hemorrhage, and other complications of prematurity. Data analyses were performed in accordance with the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: Only one randomized controlled clinical trial met selection criteria and was included in this review (Verder 1994). In this study of infants with signs of RDS, intubation and early surfactant therapy followed by extubation to nasal CPAP (NCPAP) compared with later, selective surfactant administration was associated with a lower incidence of mechanical ventilation (ventilation continuing for one hour or more after surfactant administration in the early surfactant group or initiated for respiratory insufficiency or apnea in either group [RR 0.51, 95% CI 0.32, 0.76]). A larger proportion of infants in the early surfactant group received surfactant than in the selective surfactant group [RR 1.74, 95% CI 1.30, 2.33]. The number of surfactant doses per patient was significantly greater among patients randomized to the early surfactant group [MD 0.51, 95% CI 0.32, 0.70]. Trends towards a decreased incidence of mortality, and a higher rate of patent ductus arteriosus requiring treatment were seen in the early surfactant group. There was no evidence of effect on median time in oxygen, duration of mechanical ventilation, or incidence of BPD (oxygen at 28 days). REVIEWER'S CONCLUSIONS: Early surfactant replacement therapy with extubation to NCPAP compared with later, selective surfactant replacement and continued mechanical ventilation with extubation from low ventilator support is associated with a reduced need for mechanical ventilation and increased utilization of exogenous surfactant therapy. These conclusions are based on findings from one small randomized clinical trial. Additional randomized trials are needed and are underway. PMID- 12076470 TI - Hydration for treatment of preterm labour. AB - BACKGROUND: Hydration has been proposed as a treatment for women with preterm labour. Theoretically, hydration may reduce uterine contractility by increasing uterine blood flow and by decreasing pituitary secretion of antidiuretic hormone and oxytocin. OBJECTIVES: To evaluate the effectiveness of intravenous or oral hydration to avoid preterm birth and its consequences in women with preterm labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register (January 2002), the Cochrane Controlled Trials register (The Cochrane Library, Issue 1, 2002) and bibliographies of relevant papers were searched. SELECTION CRITERIA: Randomised controlled trials, including women with a viable pregnancy less than 37 completed weeks' gestation and presenting with preterm labour, comparing intravenous or oral hydration with no treatment. The intervention might or might not be associated with bed rest. Studies comparing tocolytic drugs with intravenous fluids used in the control group as a placebo were not included in this review. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the reports, to determine if the study met the inclusion criteria and to evaluate the methodological quality. Data were extracted independently by two of the reviewers. The results were expressed as relative risks (RR) for dichotomous outcomes and weighted mean difference for continuous outcomes. MAIN RESULTS: Two studies, including a total of 228 women with preterm labour and intact membranes, compared intravenous hydration with bed rest alone. Risk of preterm delivery, before 37 weeks (relative risk (RR): 1.09; 95% confidence interval (CI): 0.71-1.68), before 34 weeks (RR: 0.72; 95% CI: 0.20 2.56) or before 32 weeks (RR: 0.76; 95% CI: 0.29-1.97), was similar between groups. Admission to neonatal intensive care unit occurred with similar frequency in both groups (RR: 0.99; 95% CI: 0.46-2.16). Cost of treatment was slightly higher (US$39) in the hydration group. This difference was not statistically significant and only includes hospital costs during a visit of less than 24 hours. No studies evaluated oral hydration. REVIEWER'S CONCLUSIONS: The data are too few to support the use of hydration as a specific treatment for women presenting with preterm labour. The two small studies available do not show any advantage of hydration compared to bed rest alone. Intravenous hydration does not seem to be beneficial, even during the period of evaluation soon after admission, in women with preterm labour. Women with evidence of dehydration may, however, benefit from the intervention. PMID- 12076471 TI - D-cycloserine for Alzheimer's disease. AB - BACKGROUND: Evidence supports a role for the NMDA receptors in learning and memory. These can be modulated by the antibiotic D-cycloserine in such a way that the effect of the excitatory transmitter substance glutamate is enhanced. A study on healthy subjects pretreated with scopolamine to mimic Alzheimer's disease showed a positive effect of D-cycloserine at low doses. OBJECTIVES: To assess the efficacy and safety of D-cycloserine in patients with Alzheimer's disease. SEARCH STRATEGY: The trials were identified from a search of the Specialized Register of the Cochrane Dementia and Cognitive Improvement Group on 14 June 2001 using the terms: cycloserine, D-cycloserine, Alzheimer*. SELECTION CRITERIA: Randomized, double-blinded and unconfounded trials comparing D-cycloserine with a control treatment. DATA COLLECTION AND ANALYSIS: Two larger and two smaller randomized controlled trials were identified. The clinical global impression scale was used in all studies and was a primary outcome measure. MAIN RESULTS: It was not possible to extract the results from the first phases of the two crossover studies and therefore the meta-analyses are based on the two parallel group 6 month studies. There was no indication of a positive effect favouring D cycloserine for the numbers showing improvement at 6 months as assessed by the Clinical Global Impression for any dose. The number of withdrawals for any reason before end of treatment at 6 months was significantly in favour of placebo (fewer withdrawals) compared with D-cycloserine for dose levels of 30 mg/day (OR 2.94, 95% CI 1.52, 5.70) and 100 mg/day (OR 3.23, 95% CI 1.67, 6.25). There was no significant difference between treatment, (2, 10, 30, 100, or 200 mg/day) and placebo for the number of withdrawals due to adverse events by six months. REVIEWER'S CONCLUSIONS: The lack of a positive effect of D-cycloserine on cognitive outcomes in controlled clinical trials with statistical power high enough to detect a clinically meaningful effect means that D-cycloserine has no place in the treatment of patients with Alzheimer's disease. PMID- 12076472 TI - Cognitive behavioural interventions for sleep problems in adults aged 60+. AB - BACKGROUND: The prevalence of sleep problems in adulthood increases with age. While not all sleep changes are pathological in later life, severe disturbances may lead to depression, cognitive impairments, deterioration of quality of life, significant stresses for carers and increased healthcare costs. The most common treatment for sleep disorders (particularly insomnia) is pharmacological. The efficacy of non-drug interventions has been suggested to be slower than pharmacological methods, but with no risk of drug-related tolerance or dependency. Cognitive and behavioural treatments for sleep problems aim to improve sleep by changing poor sleep habits, promoting better sleep hygiene practices and by challenging negative thoughts, attitudes and beliefs about sleep. OBJECTIVES: To assess the efficacy of cognitive-behavioural interventions in improving sleep quality, duration and efficiency amongst older adults (aged 60 and above). SEARCH STRATEGY: The following databases were searched: MEDLINE (1966 - October 2001); EMBASE (1980 - January 2002), CINAHL ( 1982 - January 2002; PsychINFO 1887 to 2002; The Cochrane Library (Issue 1, 2002); National Research Register (NRR [2002]). Bibliographies of existing reviews in the area, as well as of all trial reports obtained, were searched. Experts in the field were consulted. SELECTION CRITERIA: Randomised controlled trials of cognitive behavioural treatments for primary insomnia where 80% or more of participants were over 60. Participants must have been screened to exclude those with dementia and/or depression. DATA COLLECTION AND ANALYSIS: Abstracts of studies identified in searches of electronic databases were read and assessed to determine whether they might meet the inclusion criteria. Data were analysed separately depending on whether results had been obtained subjectively or objectively. MAIN RESULTS: Six trials, including 282 participants with insomnia, examined the effectiveness of cognitive-behavioural treatments (CBT) for sleep problems in this population. The final total of participants included in the meta-analysis was 224. The data suggest a mild effect of CBT for sleep problems in older adults, best demonstrated for sleep maintenance insomnia. REVIEWER'S CONCLUSIONS: When the possible side-effects of standard treatment (hypnotics) are considered, there is an argument to be made for clinical use of cognitive-behavioural treatments. Research is needed to establish the likely predictors of success with such treatments. As it may well be the case that the treatment efficacy of cognitive behavioural therapy itself is not durable, the provision of "top-up" sessions of CBT training to improve durability of effect are worthy of investigation. PMID- 12076473 TI - Physical therapies for prevention of urinary and faecal incontinence in adults. AB - BACKGROUND: Physical therapies, particularly pelvic floor muscle training, have been recommended for prevention of incontinence associated with childbearing, and prostate surgery. OBJECTIVES: To assess the effectiveness of physical therapies in preventing incontinence in adults. SEARCH STRATEGY: The Cochrane Incontinence Group trials register was searched to September 2001. Trials were also sought from the Reference Lists of relevant articles and from experts in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials in adults without incontinence symptoms that compared a physical therapy with no treatment, or any other treatment to prevent incontinence. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed eligibility, trial quality and extracted data. Data were processed as described in the Cochrane Collaboration Handbook. MAIN RESULTS: Two trials in men (155 men) and 13 trials in women (4661 women) were included. As most trials recruited regardless of continence status, and there was much heterogeneity, only a qualitative synthesis was undertaken. Three of seven trials in childbearing women reported less urinary incontinence after pelvic floor muscle training compared to control treatment three months postpartum. Two trials selected women at higher risk of postnatal incontinence. The third used an intensive training programme. Four trials did not find any difference between the groups at the primary endpoint. Two trials compared pre-prostate surgery pelvic floor muscle training with control treatment, and no difference in the occurrence of postoperative urinary incontinence was reported between the groups. REVIEWER'S CONCLUSIONS: There is insufficient evidence to determine whether physical therapies can prevent incontinence in childbearing women, or men following prostate surgery. Further, better quality research is needed. PMID- 12076474 TI - Immunosuppressive treatment for multifocal motor neuropathy. AB - BACKGROUND: Multifocal motor neuropathy is a distinct clinical entity characterised by progressive, predominantly distal, asymmetrical limb weakness and minimal sensory abnormality. The pathognomonic feature of this condition is the presence of multiple partial motor nerve conduction blocks. Controlled trials have demonstrated the efficacy of regular intravenous immunoglobulin infusions. Immunosuppressive agents have been used as primary, second-line or adjunctive agents for its treatment. This review was undertaken to identify and review systematically randomised controlled trials of immunosuppressive agents. The use of intravenous immunoglobulin will be the subject of a separate review. OBJECTIVES: To provide the best available evidence from randomised controlled trials on the role of immunosuppressive agents for the treatment of multifocal motor neuropathy. SEARCH STRATEGY: The Cochrane Neuromuscular Disease Group register was searched for all trials of multifocal motor neuropathy published, using 'multifocal motor neuropathy' OR 'chronic inflammatory demyelinating polyradiculoneuropathy' OR ' conduction block' OR ' motor neuropathy' AND 'immunosuppressive agents', 'immunosuppressants', 'corticosteroids', 'plasma exchange', 'azathioprine', 'cyclophosphamide', 'cyclosporin', 'methotrexate', and 'mycophenolate', 'immunomodulatory agents', 'interferon', 'total lymphoid irradiation' or 'bone marrow transplantation' as search terms. In addition we searched MEDLINE, EMBASE for 2000 and 2001 and CINAHL, LILACS for all years. SELECTION CRITERIA: All randomised controlled trials and quasi-randomised clinical trials in which allocation was not random but was intended to be unbiased (e.g. alternate allocation) were to have been selected. Since no such trials were discovered, all prospective and retrospective case series were included in 'background' or 'discussion' sections of the review. DATA COLLECTION AND ANALYSIS: All studies on multifocal motor neuropathy or lower motor neuron weakness with conduction block and no sensory abnormality were scrutinized for data on patients treated with any form of immunosuppressive agents besides intravenous immunoglobulin. The information on the outcome of treatment was then collated and summarised. MAIN RESULTS: We found no randomised controlled trials of any immunosuppressive agents for multifocal motor neuropathy. We summarised the results of retrospective and prospective case series in the discussion of the review. REVIEWER'S CONCLUSIONS: There are no randomised controlled trials to indicate whether immunosuppressive agents are beneficial in multifocal motor neuropathy. PMID- 12076475 TI - Rehabilitation for distal radial fractures in adults. AB - BACKGROUND: Fracture of the distal radius is a common clinical problem, particularly in older white women with osteoporosis. OBJECTIVES: To examine the evidence for effectiveness of rehabilitation intervention(s) for adults with conservatively or surgically treated distal radial fractures. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register (January 2002), the Cochrane Controlled Trials Register (The Cochrane Library, Issue 4, 2001), the Cochrane Rehabilitation and Related Therapies Field database, MEDLINE (1966 to January 2002), EMBASE (1988 to 2001 Week 50), CINAHL (1982 to December Week 2 2001), Current Controlled Trials (December 2001), AMED, PEDro, conference proceedings and reference lists of articles. SELECTION CRITERIA: Randomised or quasi-randomised clinical trials evaluating rehabilitation as part of the management of fractures of the distal radius sustained by skeletally mature patients. Rehabilitation interventions such as active and passive mobilisation exercises, and training for activities of daily living, could be used on their own or in combination, and be applied in various ways by various clinicians. DATA COLLECTION AND ANALYSIS: All trials meeting the selection criteria were independently assessed by all three reviewers for methodological quality. Data were extracted independently by two reviewers. The trials were grouped into categories relating to the main comparisons, and to when the intervention(s) commenced (for example, during or after plaster cast immobilisation). Quantitative data are presented using relative risks or mean differences together with 95 per cent confidence limits. MAIN RESULTS: Twelve trials, involving 601 mainly female and older patients, were included. Initial treatment was conservative, involving plaster cast immobilisation, in all but 20 patients whose fractures were fixed surgically. Though some trials were well conducted, others were methodologically compromised. No trial provided definitive evidence. Only very limited pooling of results from comparable trials was possible. During immobilisation, there was weak evidence of improved hand function in the short term, but not in the longer term, for early occupational therapy (1 trial), and of a lack of differences in outcome between supervised and unsupervised exercises (1 trial). Post-immobilisation, there was weak evidence of a lack of clinically significant differences in outcome in patients receiving formal rehabilitation therapy (3 trials), passive mobilisation (2 trials) or whirlpool immersion (1 trial) compared with no intervention. There was weak evidence of a short-term benefit of continuous passive motion (post external fixation) (1 trial), intermittent pneumatic compression (1 trial) and ultrasound (1 trial). There was weak evidence of better short-term hand function in patients given physiotherapy than in those given instructions for home exercises by a surgeon (1 trial). REVIEWER'S CONCLUSIONS: The available evidence from randomised trials is insufficient to establish the relative effectiveness of the various interventions used in the rehabilitation of adults with fractures of the distal radius. PMID- 12076476 TI - In vitro fertilisation for unexplained subfertility. AB - BACKGROUND: In vitro fertilisation (IVF) is now a widely accepted treatment for unexplained infertility (RCOG 1998). However, with estimated livebirth rates per cycle varying between 13% and 28%, it's effectiveness has not been rigorously evaluated in comparison with other treatments. With increasing awareness of the role of expectant management and less invasive procedures such as intrauterine insemination, concerns about multiple complications and costs associated with IVF, it is extremely important to evaluate the effectiveness of IVF against other treatment options in couples with unexplained infertility. OBJECTIVES: The aim of this review is to determine, in the context of unexplained infertility, whether IVF improves the probability of livebirth compared with 1. expectant management 2. clomiphene citrate (CC) 3. intra uterine insemination (IUI) alone 4. IUI with controlled ovarian stimulation and 5. Gamete IntraFallopian Transfer (GIFT). SEARCH STRATEGY: RCTs were identified using the search strategies developed for the Menstrual Disorders and Subfertility Group. See Review group for more information. SELECTION CRITERIA: Only randomised controlled trials were included. Livebirth rate per woman was the primary outcome of interest. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed eligibility and quality of trials. MAIN RESULTS: Nine randomised controlled trials were identified. In two we could not extract data separately for unexplained infertility cases, two were non randomised, one reported no valid rates (included in the review and not in the meta-analysis), leaving four trials for analysis. One trial compared two different interventions (IVF versus IUI with or without ovarian stimulation) and one study compared three interventions (IVF versus IUI with ovarian stimulation and GIFT). The number of trials assessing the effectiveness of IVF with the other treatments were as follows: IVF versus expectant management (one), IVF versus IUI (one), IVF versus IUI with ovarian stimulation (two) and IVF versus GIFT (three). Livebirth rate per woman was reported in two studies and three studies determined clinical pregnancy rate per woman. Multiple pregnancy rate was reported in three trials. Two studies reported ovarian hyperstimulation syndrome (OHSS) as an outcome measure. There were no comparative data for clomiphene citrate, and no comparative data on livebirth rates for expectant management or GIFT. There was no evidence of a difference in livebirth rates between IVF and IUI either without (OR 0.51, 95% CI 0.23 to 1.1) or with (OR 0.87, 95% CI 0.42 to 1.8) ovarian stimulation. There was no evidence of a difference in clinical pregnancy rates between IVF and expectant management. There was no significant difference in the clinical pregnancy rates between IVF and GIFT (OR 0.47, 95% CI 0.24 to 0.92). There was no evidence of a difference in the multiple pregnancy rates between IVF and either IUI with ovarian stimulation (OR 1.59, 95% CI 0.68 to 3.70) or GIFT (OR 0.47, 95% CI 0.08 to 0.58). Clinical heterogeneity was present among the studies included. However, there was no evidence of statistical heterogeneity, which allowed the studies to be combined for statistical analysis. REVIEWER'S CONCLUSIONS: Any effect of IVF relative to expectant management, clomiphene citrate, IUI with or without ovarian stimulation and GIFT in terms of livebirth rates for couples with unexplained subfertility remains unknown. The studies included are limited by their small sample size, so that even large differences might be hidden. Livebirth rates are seldom reported. Adverse effects such as multiple pregnancies and ovarian hyperstimulation syndrome have also not been reported in most studies. Larger trials with adequate power are warranted to establish the effectiveness of IVF in these women. Future trials should not only report rates per woman /couple but also include adverse effects and costs of the treatments compared as outcomes. Factors that have a major effect on these outcomes such as fertility treatment, female partner's age, duration of infertility and previous pregnancy history should also be considered. PMID- 12076477 TI - Kava extract for treating anxiety. AB - BACKGROUND: Synthetic anxiolytic drugs are effective for anxiety, but are often burdened with adverse events. Constraints on resources and time often render treatments such as psychological interventions impracticable. Thus, an effective and safe oral medication would be of considerable interest and a welcome addition to the therapeutic repertoire. OBJECTIVES: To systematically review the evidence from rigorous clinical trials assessing the efficacy and safety of kava extract versus placebo for the treatment of anxiety. SEARCH STRATEGY: Publications describing randomised controlled trials (RCTs) of kava extract for anxiety were sought through Medline, Embase, Biosis, AMED, CISCOM and the Cochrane Library (all from their respective inception to June 1998). The search terms used were kava, kawa, kavain, Piper methysticum and Rauschpfeffer (German common name for Piper methysticum). Manufacturers of kava preparations and experts on the subject were contacted and asked to contribute published and unpublished material. In addition, our own files were searched and the bibliographies of all of the studies identified were scanned for further trials. There were no restrictions on the language of publication. SELECTION CRITERIA: Randomized, double-blind trials of oral kava extract mono-preparations for the treatment of anxiety were included. Trials comparing kava with placebo were included. Trials assessing kava as one of several active components in a combination preparation or as a part of a combination treatment were excluded. DATA COLLECTION AND ANALYSIS: All publications were blinded prior to assessment by a person not involved in the study. Data were extracted systematically. Methodological quality of all trials was evaluated using the standard scoring system developed by Jadad and colleagues. The screening of studies, selection, data extraction and the assessment of methodological quality were performed independently by the two reviewers. Disagreements in the evaluation of individual trials were resolved through discussion. MAIN RESULTS: Seven trials met the inclusion criteria. All of the reviewed trials suggest superiority of kava extract over placebo. The meta analysis of three studies using the Hamilton Anxiety Score as a common outcome measure suggests a significant differential treatment effect in favour of kava extract (weighted mean difference: 9.7, 95% confidence interval: 3.5 - 15.8). Adverse events as reported in the reviewed trials were mild, transient and infrequent. REVIEWER'S CONCLUSIONS: The evidence presented implies that kava extract is superior compared with placebo and relatively safe as a treatment option for anxiety. These findings warrant further and more rigorous investigations into the efficacy and safety of kava extract. PMID- 12076478 TI - Bright light therapy for sleep problems in adults aged 60+. AB - BACKGROUND: The prevalence of sleep problems in adulthood increases with age. While not all sleep changes are pathological in later life, severe disturbances may lead to depression, cognitive impairments, deterioration of quality of life, significant stresses for carers and increased healthcare costs. The most common treatment for sleep disorders (particularly insomnia) is pharmacological. The efficacy of non-drug interventions has been suggested to be slower than pharmacological methods, but with no risk of drug-related tolerance or dependency. Bright light treatment involves participants sitting in front of a "light box" which emits very high (typically 10,000 lux) fluorescent light for periods of around two hours daily. The timing of this light treatment will depend on the irregular timing of the participant's sleep pattern. OBJECTIVES: To assess the efficacy of bright light therapy in improving sleep quality (sleep timing in particular) amongst adults aged 60 and above. SEARCH STRATEGY: The following databases were searched: MEDLINE (1966 - January 2001); EMBASE (1980 - January 2001), CINAHL ( 1982 - January 2001; PsychINFO 1970 to 2001; The Cochrane Library (Issue 1, 2001); National Research Register (NRR [2001]). Bibliographies of existing reviews in the area, as well as of all trial reports obtained, were searched. Experts in the field were consulted. SELECTION CRITERIA: Randomised controlled trials of bright light therapy for primary sleep problems where 80% or more of participants were over 60. Participants must have been screened to exclude those with dementia and/or depression. DATA COLLECTION AND ANALYSIS: Abstracts of studies identified in searches of electronic databases were read and assessed to determine whether they might meet the inclusion criteria. MAIN RESULTS: Reviewers found no trials on which to base conclusions for the effectiveness of this treatment. REVIEWER'S CONCLUSIONS: When the possible side effects of standard treatment (hypnotics) are considered, there is a reasonable argument to be made for clinical use of non-pharmacological treatments. In view of the promising results of bright light therapy in other populations with problems of sleep timing, further research into their effectiveness with older adults would seem justifiable. PMID- 12076479 TI - Methadone at tapered doses for the management of opioid withdrawal. AB - BACKGROUND: Despite widespread use in many countries of tapered methadone for detoxification from opiate dependence, the evidence of efficacy to prevent relapse and promote lifestyle change has not been systematically evaluated. OBJECTIVES: To determine whether tapered methadone is effective to manage withdrawal from opioids. SEARCH STRATEGY: We searched: the Cochrane Controlled Trials Register (Issue 1, 2000), MEDLINE (OVID 1966-2000), EMBASE (1980-2000); scan of reference list of relevant articles; personal communication; conference abstracts; unpublished trials from pharmaceutical industry; Internet (NIDA, Clinical Trials.org, BMJ). SELECTION CRITERIA: All randomised controlled trials which focus on the use of tapered methadone (length of treatment max 30 days) versus all other pharmacological detoxification treatments, placebo and different modalities of methadone detoxification programs for the treatment of opiate withdrawal. Trials including patients with additional diagnoses such as benzodiazepine dependence were also eligible. DATA COLLECTION AND ANALYSIS: One reviewer (LA) assessed studies for inclusion and undertook data extraction. Inclusion decisions and the overall process were confirmed by consultation between reviewers. Qualitative assessments of the methodology of eligible studies were carried out using validated checklists. Where possible analysis was carried out according to the "intention to treat" principles. MAIN RESULTS: 20 studies were included in the review, with 1357 people randomised. 10 studies compared methadone with adrenergic agonists, 7 studies compared different modalities of methadone detoxification, 2 studies compared methadone with other opioid agonists, 1 study compared methadone with chlordiazepoxide one with placebo. The conclusions of the 10 studies that compared methadone with adrenergic agonists showed a substantial clinical difference of the two treatments in terms of retention in treatment, degree of discomfort and detoxification success rates. The conclusions of the 6 studies that compare different methadone reduction schedules, showed that different types of methadone withdrawal schedule produce different responses in terms of time course of withdrawal, the severity of withdrawal response and possibly in terms of subsequent engagement with treatment. Regarding the studies that compare methadone with other opioid agonists, in Sorensen 1982 methadyl acetate performed similarly to methadone on most process and outcome measures, while in Tennant 1975, methadone reduced severity of withdrawal and had fewer drop-outs than did a propoxyphene group. In Drummond 1989, using chlordiazepoxide vs methadone, the results suggest that the two drugs had similar results in terms of overall effectiveness. San 1992 compared methadone with placebo and found more severe withdrawal and more drop outs in the placebo group. The results indicate that tapered methadone and other medications used in the included studies are effective in the treatment of the heroin withdrawal syndrome, although symptoms experienced by subjects differed according to the medication used and the program adopted. It seems that regardless of which medication is selected for heroin detoxification, the rates of subsequent heroin abstinence are about equal. This suggests that the medications are similar in terms of overall effectiveness. Improvements were achieved when other services such as counseling and other supporting services were offered contemporaneously with detoxification. REVIEWER'S CONCLUSIONS: Data from literature are hardly comparable; programs vary widely with regard to duration, design and treatment objectives, impairing the application of meta analysis. Results of many outcomes could not be summarised because they were presented either in graphical form or provided only statistical tests and p values. For most studies standard deviation for continuous variables were not provided. The studies included in this review confirm that slow tapering with temporary substitution of long acting opioids, accompanied by medical supervision and ancillary medications can reduce withdrawal severity. Nevertheless the majority of patients relapsed to heroin use. However this cannot be considered a goal for a detoxification as heroin dependence is a chronic, relapsing disorder and the goal of detoxification should be to remove or reduce dependence on heroin in a controlled and human fashion and not a treatment for heroin dependence. PMID- 12076480 TI - Dietary fibre for the prevention of colorectal adenomas and carcinomas. AB - BACKGROUND: Colorectal cancer (CRC) is a major cause of morbidity and mortality in industrialized countries. Experimental evidence has supported the hypothesis that dietary fibre may be protective for the development of CRC, although epidemiologic data have been inconclusive. OBJECTIVES: We have conducted a systematic review and meta-analysis to assess the effect of dietary fibre on the incidence or recurrence of colorectal adenomas, the incidence of CRC, and the development of adverse events. SEARCH STRATEGY: We identified randomized controlled trials from Medline, Embase, and the Cochrane Controlled Trials Register up to Oct 2001 SELECTION CRITERIA: Randomized or quasi-randomized controlled trials were assessed. The population included all subjects that had adenomatous polyps but no previous history of colorectal cancer (CRC), a documented "clean colon" at baseline and repeated visualization of the colon/rectum after at least two years of follow-up. Dietary fibre was the intervention. The primary outcomes were the number of subjects with: a) at least one adenoma, b) more than one adenoma, c) at least one adenoma greater than or equal to 1 cm or d) a new diagnosis of CRC. The secondary outcome was the number of adverse events. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data, assessed trial quality and resolved discrepancies by consensus. The outcomes were reported as relative risks (RR) and risk difference (RD) with 95% confidence intervals (CI). If statistical significance was reached, the number need to treat (NNTT) or harm (NNTH) was reported. The study data were combined with the fixed effects model if it was clinically, methodologically, and statistically reasonable. MAIN RESULTS: Five studies with 4349 subjects met the inclusion criteria. The interventions were wheat bran fibre, ispaghula husk, or a comprehensive dietary intervention with high fibre whole food sources alone or in combination. When the data were combined there was no difference between the intervention and control groups for the number of subjects with at least one adenoma [RR 1.04 (95% CI 0.95,1.13); RD 0.01 (95% CI 0.02,0.04)]. As well, the combined results for the number of subjects with more than one adenoma [RR 1.02 (95% CI 0.89,1.17), RD 0.00 (-0.02,0.03)] or at least one adenoma 1 cm or greater [RR 0.94 (95% CI 0.77,1.15), RD -0.01 (-0.02,0.01)] were not statistically significant. Other primary and secondary outcomes and subanalyses by type of fibre intervention were not statistically or clinically significant. REVIEWER'S CONCLUSIONS: There is currently no evidence from RCTs to suggest that increased dietary fibre intake will reduce the incidence or recurrence of adenomatous polyps within a two to four year period. PMID- 12076481 TI - Interventions for increasing pedestrian and cyclist visibility for the prevention of death and injuries. AB - BACKGROUND: Pedestrians and cyclists account for nearly one in three of all road users killed and seriously injured in road traffic crashes. Late detection of other road users is one of the basic driver failures responsible for collisions. Aids to improve pedestrians and cyclist visibility have been used to avert potential collisions. However, the impact of these strategies on drivers' responses, and on pedestrian and cyclist safety is not known. OBJECTIVES: 1. To quantify the effect of visibility aids vs no visibility aids, and of different visibility aids on the occurrence of pedestrian and cyclist-motor vehicle collisions and injuries. 2. To quantify the effect of visibility aids vs no visibility aids, and of different visibility aids on drivers' detection and recognition responses. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, the Injuries Group Specialised Register, MEDLINE, TRANSPORT, TRANSDOC from ECMT (European Conference of Ministers of Transport), IRRD (International Road Research Documentation), TRIS (Transportation Research Information Services), NRR (National Research Register), PsycInfo and PsycLit. SELECTION CRITERIA: 1. Randomised controlled trials and controlled before/after trials of the effect of visibility aids on the occurrence of pedestrian and cyclist-motor collisions and injuries. 2. Randomised controlled trials of the effect of visibility aids on drivers' detection and recognition responses. This included trials where the order of presentation of visibility aids was randomised or balanced using a Latin square design. DATA COLLECTION AND ANALYSIS: Two reviewers independently screened records, extracted data and assessed trial quality. MAIN RESULTS: We found no trials assessing the effect of visibility aids on the occurrence of pedestrian and cyclist-motor vehicle collisions and injuries. We identified 37 trials assessing the effect of visibility aids on drivers' responses. Fluorescent materials in yellow, red and orange colours improve detection and recognition in the daytime. For night-time visibility, lamps, flashing lights and retroreflective materials in red and yellow colours increase detection and recognition. Retroreflective materials arranged in a 'biomotion' configuration also enhance recognition. Substantial heterogeneity between and within the trials limited the possibility for meta-analysis. Summary statistics and descriptive summaries of the outcomes were presented for individual trials when appropriate. REVIEWER'S CONCLUSIONS: Visibility aids have the potential to increase visibility and enable drivers to detect pedestrians and cyclists earlier. Public acceptability of these strategies would merit further development. However, the effect of visibility aids on pedestrian and cyclist safety remains unknown. Studies which collect data on simple, meaningful outcomes are required. PMID- 12076482 TI - 'As required' medication regimens for seriously mentally ill people in hospital. AB - BACKGROUND: Drugs used to treat psychotic illnesses may take weeks to have an effect. Often, in the interim, additional, 'as required' doses of medication can be used to calm patients in psychiatric wards. 20 to 50% of people on acute psychiatric wards are written up for 'as required' doses of medication. In this situation, a doctor prescribes the frequency and upper limit of dose, and the drug is then given at the discretion of clinical staff. OBJECTIVES: To compare the effects of 'as required' medication regimens with regular regimens of medication for the treatment of psychotic symptoms or behavioural disturbance, thought to be secondary to psychotic illness. This is in addition to any regular psychotropic medication for the long-term treatment of schizophrenia or schizophrenia-like illnesses. SEARCH STRATEGY: The reviewers searched the Cochrane Schizophrenia Group's register of trials (November 2001). This register is compiled by methodical searches of BIOSIS, CINAHL, Dissertation abstracts, EMBASE, LILACS, MEDLINE, PSYNDEX, PsycINFO, RUSSMED, Sociofile and is supplemented with hand searching of relevant journals and numerous conference proceedings. SELECTION CRITERIA: Relevant randomised control trials, involving hospital inpatients with schizophrenia or schizophrenia-like illnesses, comparing any regimen of medication administered for the short term relief of behavioural disturbance, or psychotic symptoms, to be given at the discretion of ward staff ('as required', 'prn') with fixed non-discretionary patterns of drug administration of the same drug(s). This is in addition to regular psychotropic medication for the long-term treatment of schizophrenia or schizophrenia-like illnesses where prescribed. The primary outcomes of interest were levels of disturbance, severity of psychiatric symptoms, dose of medication, and adverse effects. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality rated and data extracted. For dichotomous data, relative risks (RR) would have been estimated, with the 95% confidence intervals (CI). Where possible, the number needed to treat statistic (NNT) was to have been calculated. Analysis would have been by intention-to-treat. Normal continuous data were to have been summated using the weighted mean difference (WMD). Scale data were to have been presented for only those tools which had attained pre-specified levels of quality. Tests of heterogeneity and for publication bias were to have been undertaken. MAIN RESULTS: No randomised trials comparing 'as required' medication regimens to regular regimens of the same drug were identified. REVIEWER'S CONCLUSIONS: This common current practice has no support from randomised trials. Current practice is based on clinical experience and habit rather than high quality evidence. Current practice, therefore, outside of a well designed, conducted and reported randomised trial, is therefore difficult to justify. PMID- 12076483 TI - Transcranial magnetic stimulation for treating depression. AB - BACKGROUND: Transcranial magnetic stimulation can either excite or inhibit cortical areas of the brain, depending on whether the speed of the repetitive stimulation is applied at high or low frequencies. It has been used for physiological studies and it has also been proposed as a treatment for depression. OBJECTIVES: To assess the clinical efficacy and safety of transcranial magnetic stimulation for treating depression. SEARCH STRATEGY: An electronic search was performed including the Cochrane Collaboration Depression, Neurosis and Anxiety Review Group trials register (last searched June, 2001), the Cochrane Controlled Trials Register (Issue 2, 2001), MEDLINE (1966-2001), EMBASE (1974-2001), PsycLIT (1980-2001), and bibliographies from reviewed articles. Unpublished data and grey literature were searched through personal communications with researchers. SELECTION CRITERIA: Randomised controlled trials assessing the therapeutic efficacy and safety of transcranial magnetic stimulation for depression. DATA COLLECTION AND ANALYSIS: All reviewers independently extracted the information and verified it by cross-checking. Disagreements were resolved through discussion. Continuous data: When similar studies were grouped, the overall standardised mean difference was calculated under a fixed effect model weighted by the inverse variance method with 95% confidence intervals. (In the presence of statistical heterogeneity, a random effects model was to be used.) MAIN RESULTS: Sixteen trials were included in the review and fourteen contained data in a suitable form for quantitative analysis. Most comparisons did not show differences between rTMS and other interventions. No difference was seen between rTMS and sham TMS using the Beck Depression Inventory or the Hamilton Depression Rating Scale, except for one time period (after two weeks of treatment) for left dorsolateral prefrontal cortex and high frequency; and also for right dorsolateral prefrontal cortex and low frequency, both in favour of rTMS and both using the Hamilton scale. Comparison of rTMS (left dorsolateral prefrontal cortex and high frequency) with electroconvulsive therapy showed no difference except for psychotic patients after two weeks treatment, using the Hamilton scale, which indicated that electroconvulsive therapy was more effective than rTMS. REVIEWER'S CONCLUSIONS: The information in this review suggests that there is no strong evidence for benefit from using transcranial magnetic stimulation to treat depression, although the small sample sizes do not exclude the possibility of benefit. PMID- 12076484 TI - Antiretrovirals for reducing the risk of mother-to-child transmission of HIV infection. AB - BACKGROUND: At the end of 2000 it was estimated that over 36 million people were living with the human immunodeficiency virus (HIV). This includes 1.4 million children less than 15 years of age. This is one of several reviews assessing the available evidence for preventing mother-to-child transmission of HIV infection. The other reviews will address other interventions, including Caesarean section, breast feeding, vaginal lavage and vitamin A supplementation. OBJECTIVES: To assess which antiretroviral therapies may be effective in decreasing the risk of mother-to-child transmission of HIV infection as well as their effect on neonatal and maternal mortality and morbidity. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register. We also searched conference abstracts from the International AIDS Conferences and Conference on Retroviruses and Opportunistic Infections. SELECTION CRITERIA: Randomised trials comparing any antiretroviral therapy aimed at decreasing the risk of mother-to-child transmission of HIV infection with placebo or no treatment, or any two or more antiretroviral therapies or regimens aimed at decreasing the risk of mother-to-child transmission of HIV infection. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Zidovudine monotherapy Based on four trials any zidovudine regimen versus placebo significantly reduces the risk of mother-to child transmission (Peto odds ratio (OR) 0.46, 95% confidence interval (CI) 0.35 to 0.60). Zidovudine also appears to decrease the risk of infant death within the first year of birth (OR 0.57, 95% CI 0.38 to 0.85) and the risk of maternal death (OR 0.32, 95% CI 0.16 to 0.66). There is no evidence that zidovudine influences the incidence of premature delivery (OR 0.86, 95% CI 0.57 to 1.29) or low birth weight (OR 0.74, 95% CI 0.53 to 1.04). The risk of transmission using a 'short short' course of zidovudine (from 35 weeks in pregnancy for the mother and for the baby until 3 days old) was higher than the risk using a 'long-long' course (from 28 weeks in pregnancy for the mother and for the baby until 6 weeks old), (OR 2.33, 95% CI 1.16 to 4.68). However, the effectiveness of the 'long-short' course (from 28 weeks in pregnancy for the mother and for the baby until 3 days old) and the 'short-long' course (from 35 weeks in pregnancy for the mother and for the baby until 6 weeks old) did not differ from that of the 'long-long' course. Nevirapine One large randomised controlled trial demonstrates that nevirapine given to mothers as a single dose at the onset of labour and to babies as a single dose within 72 hours of birth is more effective than an intrapartum and post-partum regimen of zidovudine (OR 0.51, 95% CI 0.33 to 0.79). When nevirapine is given to mothers already receiving standard antiretroviral therapy, however, there appears to be no additional advantage (OR 1.10, 95% CI 0.42 to 2.86). Combination Therapy Preliminary findings of the effect of combination therapy using zidovudine and lamivudine (3TC) suggest a decrease in the risk of transmission when the combination is given during the antenatal and intrapartum period or during the intrapartum and postpartum period compared with placebo. There is no evidence that intrapartum zidovudine and lamivudine alone are sufficient to decrease the risk of transmission compared with placebo. REVIEWER'S CONCLUSIONS: Implications for practice The randomised trials included in this review provide evidence that short course zidovudine and single-dose nevirapine are effective therapies for reducing mother-to-child transmission of HIV. The challenge for low and middle income countries will be to institute this therapy in practice. In industrialised countries practice has already moved on from the current evidence and combination antiretroviral therapy aimed primarily at preventing disease progression in the mother is the standard of care. Implications for research The potential value of nevirapine used for longer durations in breastfeeding populations should be considered as it may further reduce the risk of mother-to-child transmission, particularly if combined with early weaning. On-going evaluation of combination antiretroviral therapy is essential and will have an immediate benefit for countries with the resources to adopt such treatment. The search for effective, affordable, safe and acceptable alternatives to antiretroviral therapy for reducing mother-to-child transmission in resource poor countries should remain on the research agenda. PMID- 12076485 TI - Humidity control for chronic asthma. AB - BACKGROUND: Humidity control measures in the home environment of patients with asthma have been recommended, however there is no consensus about the usefulness of these measures. OBJECTIVES: To study the effect of dehumidification of the home environment on asthma control. SEARCH STRATEGY: A search of the clinical trials registers of the Cochrane Collaboration and Cochrane Airways Group using search terms for asthma and [humid* OR water vapour OR water vapor* OR water vapour* OR water-vapor*]. SELECTION CRITERIA: Randomized controlled trials on the use of humidity control measures in the home environment of patients with asthma were evaluated for inclusion. Only one trial could be included. DATA COLLECTION AND ANALYSIS: Data was extracted using a predesigned data extraction form. No data was available for entering into RevMan for analysis. MAIN RESULTS: The included trial using mechanical ventilation with or without high efficiency vacuum cleaners did not show any clinical benefit to asthma patients. There was a decline in the house dust mite count and the antigen level. This open trial had a low sample size. REVIEWER'S CONCLUSIONS: There is a need for studying the health benefits of dehumidification by a double blind randomized controlled trial with adequate sample size measuring clinical outcomes in patients of asthma. PMID- 12076486 TI - Cardioselective beta-blockers for chronic obstructive pulmonary disease. AB - BACKGROUND: Beta-blocker therapy has a proven mortality benefit in patients with hypertension, heart failure and coronary artery disease, as well as during the perioperative period. These drugs have traditionally been considered contraindicated in patients with chronic obstructive pulmonary disease (COPD). OBJECTIVES: To assess the effect of cardioselective beta-blockers on respiratory function of patients with COPD. SEARCH STRATEGY: A comprehensive search of EMBASE, MEDLINE and CINAHL was performed using the Cochrane Airways Group registry to identify randomised blinded controlled trials from 1966 to May 2001. The search was completed using the terms: asthma*, bronchial hyperreactivity*, respiratory sounds*, wheez*, obstructive lung disease* or obstructive airway disease*, and adrenergic antagonist*, sympatholytic* or adrenergic receptor block*. We did not exclude trials on the basis of language. SELECTION CRITERIA: Randomised, blinded, controlled trials of single dose or longer duration that studied the effects of cardioselective beta-blockers on the forced expiratory volume in 1 second (FEV1) or symptoms in patients with COPD. DATA COLLECTION AND ANALYSIS: Two independent reviewers extracted data from the selected articles, reconciling differences by consensus. Two interventions studied were the administration of beta-blocker, given either as a single dose or for longer duration, and the use of beta2-agonist given after the study drug. MAIN RESULTS: Eleven studies of single-dose treatment and 8 of treatment for longer duration, ranging from 2 days to 12 weeks, met selection criteria. Cardioselective beta blockers produced no statistically significant change in FEV1 or respiratory symptoms compared to placebo, given as a single dose (Weighted Mean Difference 2.05% [95% Confidence interval, -6.05 to 1.96%]) or for longer duration (WMD 2.55% [95% CI, -5.94 to 0.84]), and did not significantly affect the FEV1 treatment response to beta2-agonists. Exacerbations and hospitalizations were recorded in all trials, but none occurred during the periods of study, in either group. A subgroup analysis revealed no significant change in results for those participants with severe chronic airways obstruction or for those with a reversible obstructive component. REVIEWER'S CONCLUSIONS: The available evidence suggests that cardioselective beta-blockers, given to patients with COPD do not produce a significant short-term reduction in airway function or in the incidence of COPD exacerbations. However, the trials were small and of short duration. Given their demonstrated benefit in conditions such as heart failure, coronary artery disease and hypertension, cardioselective beta-blockers should be considered for patients with COPD, but administered with careful monitoring since data concerning long term administration and their effects during exacerbations are not available. PMID- 12076487 TI - Heliox for treatment of exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND: Due to its low density properties, helium-oxygen mixtures have the potential to decrease the work of breathing and possibly avoid the need for intubation and mechanical ventilation in patients with respiratory failure. OBJECTIVES: To determine the effect of the addition of helium/oxygen mixtures (heliox) to standard medical care during ventilated and nonventilated acute exacerbations of COPD. SEARCH STRATEGY: Randomized controlled trials were identified from the Cochrane Airways Review Group asthma Register. Primary authors and experts were contacted. References from included and excluded studies, known reviews and texts were also searched. SELECTION CRITERIA: Studies were selected for inclusion if they compared treatment with heliox to placebo (oxygen or air) in randomized controlled trials in adults with an exacerbation of COPD. DATA COLLECTION AND ANALYSIS: Data from all trials were combined using the Review Manager (version 4.1). We planned to perform: 1) random effects weighted mean difference (WMD), with 95% confidence intervals (95% CI), 2) Homogeneity of effect sizes with the Dersimonian and Laird method with p<0.1 as the cut point for significance, and 3) sensitivity analysis on different helium-oxygen mixtures (80/20 vs 70/30), and 4) methodological quality (Jadad score >2 vs. <3). MAIN RESULTS: Four studies, all published between 1997 and 2000 met the inclusion criteria. Two studies compared heliox-oxygen vs. air-oxygen in decompensated COPD patients who were not ventilated. One study was performed in mechanical ventilated patients and one in patients undergoing noninvasive pressure support (NIPSV) ventilated patients. Data could be obtained for only two of the studies. One was a randomized crossover study of 70:30 helium-oxygen vs air-oxygen that involved nineteen patients with acute severe COPD, hospitalized in an intensive care unit for NIPSV. In the patients receiving heliox, arterial PCO2 fell more; WMD 0.8 kPa (95% CI 0.26, -1.34). The second was a randomized trial involved 47 patients with acute COPD, who presented to an Emergency Department randomized to receive updraft nebulization of albuterol and ipratropium bromide using 80% helium and 20% oxygen or compressed air as the driving gas. Treatments were administered at 0, 20, 40, and 120 minutes after randomization. There were no significant differences in the change of FEV1 and FVC between the two groups by either the 1 or 2 hours point. although a small improvement in FEF 25-75 was significantly greater in the heliox group than in the air group. REVIEWER'S CONCLUSIONS: There is currently insufficient evidence to support the use of helium-oxygen mixtures to treat acute exacerbations of COPD in either ventilated or nonventilated patients. Suitably designed randomised controlled trials with the endpoint being the avoidance of mechanical ventilation may be justified. PMID- 12076488 TI - Oral treatments for fungal infections of the skin of the foot. AB - BACKGROUND: About 15% of the population have fungal infections of the feet (tinea pedis or athlete's foot). Whilst there are many clinical presentations of tinea pedis the most common are between the toes (interdigital) and on the soles, heels and sides of the foot (plantar) which is known as moccasin foot. Once acquired the infection can spread to other sites including the nails, which can be a source of reinfection. Oral therapy is usually used for chronic conditions or when topical treatment has failed. OBJECTIVES: To assess the effects and costs of oral treatments for fungal infections of the skin of the foot (tinea pedis). SEARCH STRATEGY: Randomised controlled trials were identified from MEDLINE, EMBASE and CINAHL from the beginning of these databases to January 2000. We also searched the Cochrane Controlled trials Register (Cochrane Library issue 1, 2000) the Science Citation Index, BIOSIS, CAB-Health, Health star and Economic databases. Bibliographies were searched, podiatry journals hand searched and the pharmaceutical industry and schools of podiatry contacted. SELECTION CRITERIA: Randomised controlled trials including participants who have a clinically diagnosed tinea pedis, confirmed by microscopy and growth of dermatophytes in culture. DATA COLLECTION AND ANALYSIS: Study selection was done by two independent reviewers. Methodological quality assessment and data collection was also assessed by two independent reviewers. MAIN RESULTS: Twelve trials, involving 700 participants, were included. The two trials comparing terbinafine and griseofulvin produced a pooled risk difference of 52% (95% confidence intervals 33% to 71%) in favour of terbinafine's ability to cure infection. No significant difference was detected between terbinafine and itraconazole; fluconazole and either itraconazole and ketoconazole; or between griseofulvin and ketoconazole, although the trials were generally small. Two trials showed that terbinafine and itraconazole were effective compared with placebo. Adverse effects were reported for all drugs, with gastrointestinal effects most commonly reported. REVIEWER'S CONCLUSIONS: The evidence suggests that terbinafine is more effective than griseofulvin and that terbinafine and itraconazole are more effective than no treatment. PMID- 12076489 TI - Cognitive rehabilitation for spatial neglect following stroke. AB - BACKGROUND: Unilateral spatial neglect is a failure to attend to one side of space. Various strategies have been used to treat these problems but evidence of their benefit has been lacking. OBJECTIVES: To determine the effects of cognitive rehabilitation for spatial neglect following stroke as measured on impairment and disability level assessments, and destination on discharge from hospital. To determine whether any effects persist at follow-up assessment. SEARCH STRATEGY: We searched the Cochrane Stroke Group Trials Register (last searched February 2001), MEDLINE (1966-December 2000), EMBASE (1980-February 2001), CINAHL (1983 January 2001), PSYCLIT and CLINPSYCH (1974-February 2001). We handsearched relevant journals, screened reference lists from relevant articles and tracked citations using SCISEARCH. SELECTION CRITERIA: Controlled trials of cognitive rehabilitation for spatial neglect in stroke. Studies with mixed patient groups were excluded unless more than 75% of their sample were stroke patients or separate stroke data were available for stroke patients. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials, extracted data, and assessed trial quality. MAIN RESULTS: We included 15 studies with 400 participants. A large number of different outcome measures were reported. Only six studies included a measure of disability and only four (111 participants) investigated persisting effects on any outcome. There was evidence that cognitive rehabilitation resulted in significant and persisting improvements in performance on impairment level assessments, although this varied depending on the test used. There was insufficient evidence to confirm or exclude an effect of cognitive rehabilitation at the level of disability or on destination following discharge from hospital. REVIEWER'S CONCLUSIONS: There is some evidence that cognitive rehabilitation for spatial neglect improves performance on some impairment level tests but its effect on disability is unclear. Further well-designed RCTs are warranted as well as basic research to develop valid outcome measures. PMID- 12076492 TI - Advice to stay active as a single treatment for low back pain and sciatica. AB - BACKGROUND: Low back pain is one of the most common conditions managed in primary care. Restricted activity, rest, and symptomatic analgesics are the most commonly prescribed treatment for low back pain and sciatica. OBJECTIVES: To assess the effects of advice to stay active as single treatment for patients with low back pain. SEARCH STRATEGY: Computerised searches in MEDLINE, EMBASE, Sport, The Cochrane Controlled Trials Register, Musculoskeletal Group's Trials Register and Scisearch, and scanning of reference lists from relevant articles were undertaken. Relevant studies were also traced by contacting authors. Date of the most recent searches: December 1998. SELECTION CRITERIA: We included all randomised trials or quasi-randomised trials where the study population consisted of adult patients with low back pain or sciatica, in which one comparison group was advised to stay active. The main outcomes of interest were pain, functional status, recovery and return to work. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion, assessed the validity of included trials and extracted data. Investigators were contacted to collect missing data or for clarification when needed. MAIN RESULTS: Four trials, with a total of 491 patients, were included. Advice to stay active was compared to advice to rest in bed in all trials. Two trials were assessed to have low risk of bias and two to have moderate to high risk of bias. The results were heterogeneous. Results from one high quality trial of patients with acute simple LBP found small differences in functional status [Weighted Mean Difference (on a 0-100 scale) 6.0 (95% CI: 1.5, 10.5)] and length of sick leave [WMD 3.4 days (95% CI: 1.6, 5.2)] in favour of staying active compared to advice to stay in bed for two days. The other high quality trial compared advice to stay active with advice to rest in bed for 14 days for patients with sciatic syndrome, and found no differences between the groups. One of the high quality trials also compared advice to stay active with exercises for patients with acute simple LBP, and found improvement in functional status and reduction in sick leave in favour of advice to stay active. REVIEWER'S CONCLUSIONS: The best available evidence suggests that advice to stay active alone has small beneficial effects for patients with acute simple low back pain, and little or no effect for patients with sciatica. There is no evidence that advice to stay active is harmful for either acute low back pain or sciatica. If there is no major difference between advice to stay active and advice to rest in bed, and there is potential harmful effects of prolonged bed rest, then it is reasonable to advise people with acute low back pain and sciatica to stay active. These conclusions are based on single trials. PMID- 12076491 TI - Pre-operative autologous donation for minimising perioperative allogeneic blood transfusion. AB - BACKGROUND: Public concerns regarding the safety of transfused blood have prompted re-consideration of the indications for the transfusion of allogeneic red cells (blood from an unrelated donor), and a range of techniques designed to minimise transfusion requirements. OBJECTIVES: To examine the evidence for the efficacy of pre-operative autologous blood donation (PAD) in reducing the need for peri-operative allogeneic red blood cell (RBC) transfusion. SEARCH STRATEGY: Articles were identified by: computer searches of OVID MEDLINE, EMBASE, and Current Contents (to March 2001) and web sites of international health technology assessment agencies (to January 2001). References in the identified trials were checked and authors contacted to identify additional studies. SELECTION CRITERIA: Randomised controlled trials with a concurrent control group in which adult patients, scheduled for non-urgent surgery, were randomised to PAD, or to a control group who did not receive the intervention. DATA COLLECTION AND ANALYSIS: Trial quality was assessed using criteria proposed by Schulz et al (1995) and Jadad et al (1996). The principle outcomes were: the number of patients exposed to allogeneic red blood cells, and the amount of blood transfused. Other clinical outcomes are detailed in the review. MAIN RESULTS: Overall PAD reduced the risk of receiving an allogeneic blood transfusion by a relative 63% (RR=0.37: 95%CI:0.26,0.54). The absolute reduction in risk of allogeneic transfusion was 43.8% (RD=-0.438: 95%CI: -0.607,-0.268). In contrast the results show that the risk of receiving any blood transfusion (allogeneic and/or autologous) is actually increased by pre-operative autologous blood donation (RR=1.29: 95%CI: 1.12,1.48). Trials were unblinded and allocation concealment was not described in 87.5% of the trials. REVIEWER'S CONCLUSIONS: Although the trials of PAD showed a reduction in the need for allogeneic blood the methodological quality of the trials was poor and the overall transfusion rates (allogeneic and/or autologous) in these trials were high, and were increased by recruitment into the PAD arms of the trials. This raises questions about the true benefit of PAD. In the absence of large, high quality trials using clinical endpoints, it is not possible to say whether the benefits of PAD outweigh the harms. PMID- 12076493 TI - Bromocriptine/levodopa combined versus levodopa alone for early Parkinson's disease. AB - BACKGROUND: Drugs that mimic dopamine such as bromocriptine were introduced as monotherapy or in a combination with LD in the hope that this approach would prevent or delay the onset of motor complications in patients with Parkinson's disease (PD). However, hitherto, the role of bromocriptine (BR) in this issue has remained controversial. The present study is a systematic review of all randomised controlled trials of bromocriptine/levodopa (BR/LD) combination therapy compared with levodopa (LD) monotherapy in PD. OBJECTIVES: To assess the efficacy and safety of BR/LD combination therapy in delaying the onset of motor complications associated with LD monotherapy in patients with PD. SEARCH STRATEGY: Sources including the Cochrane Library, the search strategy of the Movement Disorders Group (includes computerised searches of MEDLINE and EMBASE and hand searching of appropriate neurology journals), reference lists of the reviews found by the MEDLINE and EMBASE search-strategy, Sandoz -now Novartis- (manufacturer of BR), PPD Pharmaco, symposia reports, PD handbooks, contacts with colleagues who had co-ordinated trials on BR and reference lists of all included studies were used to identify randomised controlled trials (RCTs) of interest. SELECTION CRITERIA: Randomised trials were eligible for inclusion if they evaluated the efficacy of BR/LD combination therapy for delaying the onset of motor complications compared to LD monotherapy in PD patients. Outcome measures that were evaluated included occurrence and severity of motor complications, scores on impairment and disability, and the occurrence of side effects and dropouts. DATA COLLECTION AND ANALYSIS: To determine the feasibility of a quantitative systematic review two independent reviewers evaluated the methodological quality of identified trials. MAIN RESULTS: The methodological quality of five trials showed important shortcomings. All studies failed to adequately describe randomisation procedures. Only two were carried out according to a double-blind design. Differences between studies concerning the mean age of the patients, the BR titration phase, the maximum achieved daily dose of LD (62.5 1000 mg) and BR (5-50 mg), and the applied outcomes were found. Our results show no evidence of consistent differences concerning the occurrence and severity of motor complications, scores of impairment and disability and the occurrence of side effects between both treatment groups. REVIEWER'S CONCLUSIONS: This systematic review found no evidence in support of early bromocriptine/levodopa combination therapy as a strategy to prevent or delay the onset of motor complications in the treatment of PD. PMID- 12076494 TI - Bladder neck needle suspension for urinary incontinence in women. AB - BACKGROUND: Bladder neck needle suspension is an operation traditionally used for moderate or severe stress urinary incontinence in women. About a third of adult women experience some urinary incontinence, and about a third of them have moderate or severe symptoms. OBJECTIVES: To determine the effects of needle suspension on stress or mixed urinary incontinence in comparison with other management options. SEARCH STRATEGY: We searched the Cochrane Incontinence Group specialised register and reference lists of relevant articles. Date of last search: December 2001. SELECTION CRITERIA: Randomised or quasi-randomised trials that included needle suspension for the treatment of urinary incontinence. DATA COLLECTION AND ANALYSIS: Trials were assessed and data extracted independently by at least two reviewers. One trial investigator was contacted for additional information. MAIN RESULTS: Eight trials were identified which included 327 women having six different types of needle suspension procedures and 407 who received comparison interventions. Needle suspensions were more likely to fail than open abdominal retropubic suspension (higher subjective failure rate after the first year (70/267, 26% failed vs 33/243, 14% failed after open abdominal retropubic suspension: RR 2.10, 95% CI 1.45 to 3.04) and there were more peri-operative complications (36/75, 48% vs 23/77, 30%; RR 1.51, 95% CI 1.06 to 2.15), but there were no significant differences for other outcome measures. This effect was confined to women with primary incontinence: too few women with recurrent incontinence were studied to draw conclusions about the effects of secondary operations. Needle suspensions may be as effective as anterior vaginal repair (46/128, 36% failed after needles vs 50/129, 39% after anterior repair; RR 0.93, 95% CI 0.68 to 1.26) but there was little information about morbidity. Data for comparison with suburethral slings were inconclusive because they came from a small and atypical population. No trials compared needle suspensions with conservative management, peri-urethral injections or sham or laparoscopic surgery. REVIEWER'S CONCLUSIONS: Bladder neck needle suspension surgery is probably not as good as open abdominal retropubic suspension for the treatment of primary genuine stress urinary incontinence in terms of lower cure rates and higher morbidity. However, the reliability of the evidence was limited by poor quality and small trials. There was not enough information to comment on secondary procedures or on comparisons with suburethral sling operations. Although cure rates were similar after needle suspension compared with after anterior vaginal repair, the data were insufficient to be reliable and inadequate to compare morbidity. PMID- 12076495 TI - Simple behavioural and physical interventions for nocturnal enuresis in children. AB - BACKGROUND: Nocturnal enuresis (bedwetting) is a socially disruptive and stressful condition which affects around 15-20% of five year olds, and up to 2% of young adults. Although there is a high rate of spontaneous remission, the social, emotional and psychological costs can be great. Simple behavioural methods of treating bedwetting include reward systems such as star charts given for dry nights, lifting or waking the children at night to urinate, retention control training to enlarge bladder capacity (bladder training) and fluid restriction. OBJECTIVES: To assess the effects of simple behavioural interventions on nocturnal enuresis in children, and to compare these with other interventions. SEARCH STRATEGY: We searched the Cochrane Incontinence Group trials register. The reference list of a previous version of this review was also searched. Date of the most recent search: December 2001. SELECTION CRITERIA: All randomised or quasi-randomised trials of simple behavioural interventions for nocturnal enuresis in children up to the age of 16. Trials focused solely on daytime wetting were excluded. DATA COLLECTION AND ANALYSIS: In a previous version of this review, trials up to 1997 were independently assessed by two reviewers. Previous trials were re-assessed, and further trials were assessed for quality, and data were extracted by another reviewer. MAIN RESULTS: Twelve reliable trials met the inclusion criteria, involving 748 children of whom 365 received a simple behavioural intervention. However, within each comparison each outcome was addressed by single trials only, precluding meta-analysis. In single small trials, reward systems (e.g. star charts), lifting and waking were each associated with significantly fewer wet nights, higher cure rates and lower relapse rates compared to controls. There was not enough evidence to evaluate retention control training (bladder training), whether compared with controls or dry bed training, or used as a supplement to alarms, or versus desmopressin. Cognitive therapy may have lower failure and relapse rates than star charts, but this finding was based on one small trial only. One small trial of poor quality suggested that star charts were initially less successful than amitriptyline but this difference did not persist after the treatments stopped. REVIEWER'S CONCLUSIONS: Simple behavioural methods may be effective for some children, but further trials are needed, in particular in comparison with treatments known to be effective, such as desmopressin, tricyclic drugs and alarms. However, simple methods could be tried as first line therapy before considering alarms or drugs, because these alternative treatments may be more demanding and may have adverse effects. PMID- 12076496 TI - Advice on low-fat diets for obesity. AB - BACKGROUND: Overweight and obesity are global health problems contributing to an ever increasing noncommunicable disease burden. Calorie restriction can achieve short-term weight loss but the weight loss has not been shown to be sustainable in the long-term. An alternative approach to calorie restriction is to lower the fat content of the diet. However, the long-term effects of fat-restricted diets on weight loss have not been established. OBJECTIVES: To assess the effects of advice on low-fat diets as a means of achieving sustained weight loss, using all available randomised clinical trials. This review focused primarily on participants who were overweight or clinically obese and were dieting for the purpose of weight reduction. Since we were particularly interested in the ability of participants to sustain weight loss over a longer period of time, we focused on studies of 'free living' men and women who were given dietary advice rather than provision of food or money to purchase food. SEARCH STRATEGY: We searched the Cochrane Library (issue 2, 2001), MEDLINE (up to February 20002), and EMBASE (up to February 20002). We also searched the Science Citation Index (up to January 2001) and bibliographies of studies identified. Date of latest search: February 2002. SELECTION CRITERIA: Trials were included if they fulfilled the following criteria: 1) they were randomised controlled clinical trials of low-fat diets versus other weight-reducing diets, 2) the primary purpose of the study was weight loss, 3) participants were followed for at least six months, 4) the study participants were adults (18 years or older) who were overweight or obese (BMI >25 kg/m2) at baseline. Studies including pregnant women or patients with serious medical conditions were excluded. Two people independently applied the inclusion criteria to the studies identified. Disagreement was resolved by discussion or by intervention of a third party. DATA COLLECTION AND ANALYSIS: Data were extracted by three independent reviewers and meta-analysis performed using a random effects model. Weighted mean differences of weight loss were calculated for treatment and control groups at 6, 12 and 18 months. MAIN RESULTS: Four studies were included at the six month follow-up, five studies at the 12 month follow-up and three studies at the 18 month follow-up. There was no significant difference in weight loss between the two groups at six months (WMD 1.7 kg, 95% CI -1.4 to 4.8 kg). The weighted sum of weight loss in the low fat group was -5.08 kg (95% CI -5.9 to -4.3 kg) and in the control group was -6.5 kg, (95% CI -7.3 to -5.7 kg). There was no significant difference in weight loss between the two groups at 12 months (WMD 1.1 kg, 95% CI -1.6 to 3.8 kg). The weighted sum of weight loss in the low fat group was -2.3 kg (95% CI -3.2 to -1.4 kg) and in the control group was -3.4 kg (95% CI -4.2 to -2.6 kg). There was no significant difference in weight loss between the two groups at 18 months (WMD 3.7 kg, 95% CI - 1.8 to 9.2). The weighted sum of weight loss in the control group was -2.3 kg (95% CI -3.5 to -1.2 kg) and in the low fat group there was a weight gain of 0.1 kg (95% CI -0.8 to 1 kg). There was significant heterogeneity in the results for weight loss at six months and 12 months. Apart from one study which showed a slight but statistically significant difference in total cholesterol in the low fat group at one year follow-up, there were no significant differences between the dietary groups for other outcome measures such as serum lipids, blood pressure and fasting plasma glucose. Studies measuring other factors such as perceived wellness and quality of life reported conflicting results. REVIEWER'S CONCLUSIONS: The review suggests that fat-restricted diets are no better than calorie restricted diets in achieving long term weight loss in overweight or obese people. Overall, participants lost slightly more weight on the control diets but this was not significantly different from the weight loss achieved through dietary fat restriction and was so small as to be clinically insignificant. PMID- 12076497 TI - Radiotherapy for early breast cancer. AB - BACKGROUND: The long-term effects of radiotherapy on mortality from breast cancer and other causes remain uncertain. OBJECTIVES: In this report the Early Breast Cancer Trialists' Collaborative Group present their systematic overview of treatment with radiotherapy. SEARCH STRATEGY: Trial identification procedures for the EBCTCG overviews have been described elsewhere. See under "EBCTCG" in the Breast Cancer Group module. SELECTION CRITERIA: A meta-analysis was done of 10 year and 20-year results from 40 unconfounded randomised trials of radiotherapy for early breast cancer. Radiotherapy fields generally included not only chest wall (or breast) but also axillary, supraclavicular, and internal mammary nodes. DATA COLLECTION AND ANALYSIS: Data collection involved central review of individual patient data on recurrence and cause-specific mortality from 20,000 women. MAIN RESULTS: A reduction of approximately two-thirds in local recurrence was seen in all trials, largely independent of the type of patient or type of radiotherapy (8.8% vs 27.2% local recurrence by year 10). Hence, to assess effects on breast cancer mortality of substantially better local control, results from all trials were combined. Breast cancer mortality was reduced (2p=0.0001) but other, particularly vascular, mortality was increased (2p=0.0003), and overall 20-year survival was 37.1% with radiotherapy versus 35.9% control (2p=0.06). There was little effect on early deaths, but logrank analyses of later deaths indicate that, on average after year 2, radiotherapy reduced annual mortality rates from breast cancer by 13.2% (SE 2.5) but increased those from other causes by 21.2% (SE 5.4). Nodal status, age, and decade of follow-up strongly affected the ratio of breast cancer mortality to other mortality, and hence affected the ratio of absolute benefit to absolute hazard from these proportional changes in mortality. REVIEWER'S CONCLUSIONS: Radiotherapy regimens able to produce the two-thirds reduction in local recurrence seen in these trials, but without long-term hazard, would be expected to produce an absolute increase in 20-year survival of about 2-4% (except for women at particularly low risk of local recurrence). The average hazard seen in these trials would, however, reduce this 20-year survival benefit in young women and reverse it in older women. PMID- 12076498 TI - Indomethacin for the treatment of Alzheimer's disease patients. AB - BACKGROUND: Inflammatory processes involving cytokines, prostaglandins, free radicals and glial cells have been implicated in the pathogenesis of Alzheimer's disease. Non-steroidal anti-inflammatory drugs such as indomethacin attenuate inflammatory reactions. Hence, there may be a role for some of these drugs in the treatment of Alzheimer's disease. OBJECTIVES: To examine the efficacy of indomethacin in the treatment of patients suffering from Alzheimer's disease. SEARCH STRATEGY: The trials were identified from a search of the Specialized Register of the Cochrane Dementia and Cognitive Improvement Group (which contains records from many different medical and trials databases) on 14 June 2001 using the terms indomethacin and indome* and NSAIDS. In addition two independent reviewers systematically searched relevant computerized databases and Internet sites. This was supplemented by hand searching and additional references sought from selected papers. SELECTION CRITERIA: Single or multi-centre placebo controlled randomized trials examining the efficacy of indomethacin in patients diagnosed with Alzheimer's disease were eligible for selection for this review. Using a standard extraction form, inclusion/exclusion criteria were set to ensure design quality and lack of bias of all trials included. DATA COLLECTION AND ANALYSIS: Data were collected independently by two reviewers and any discrepancies were subject to discussion. Corresponding authors were contacted for any missing data needed for statistical analysis. MAIN RESULTS: Only one study was selected for this review (~~Rogers 1993~~). We detected no statistically significant difference between indomethacin treatment and placebo for the individual cognitive tests, Mini Mental State Examination (MMSE), Alzheimer's Disease Assessment Scale ( ADAS), Boston Naming Test (BNT) and Token Test (TK). Dropouts and death rate were the only reported results that were amenable to evaluation. The dropout rate was higher in the indomethacin group (10/24) than in the control group (6/20). Gastrointestinal adverse events were more prevalent in the treatment group (5/24 compared with 1/20 in control group). There was no statistically significant difference in death rate between the two groups (p=0.9). REVIEWER'S CONCLUSIONS: On the basis of this one trial and subsequent analysis of data as reported by the authors, indomethacin cannot be recommended for the treatment of mild to moderate severity Alzheimer's disease. At doses of 100-150 mg daily, serious side effects will limit its use. PMID- 12076499 TI - Paracetamol for treating fever in children. AB - BACKGROUND: Paracetamol (acetaminophen) is widely used for treating fever in children. Like ibuprofen, aspirin, and physical methods (such as fanning), paracetamol aims to provide relief from symptoms and prevent febrile convulsions. Uncertainty exists about the benefits of using it to treat fever in children. OBJECTIVES: To assess the effects of paracetamol for treating fever in children in relation to fever clearance time, febrile convulsions, and resolution of associated symptoms. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group specialized trials register (November 2001), The Cochrane Controlled Trials Register (The Cochrane Library Issue 4, 2001), MEDLINE (1966 to November 2001), EMBASE (1988 to November 2001), LILACS (2001, 40a Edition CD ROM), Science Citation Index (November 2001), and reference lists of articles. We also contacted researchers in the field. SELECTION CRITERIA: Randomized and quasi randomized trials of children with fever from infections comparing: (1) paracetamol with placebo or no treatment; and (2) paracetamol with physical cooling methods (eg, sponging, bathing, or fanning). The primary outcomes were fever clearance time and febrile convulsion. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data on methods, types of participants, interventions, and outcomes. The meta-analysis was conducted using Relative Risk with 95% confidence intervals for discrete variables, and weighted mean differences for continuous outcomes. MAIN RESULTS: 12 trials (n = 1509 participants) met the inclusion criteria. Outcomes varied between trials. No data were available on the primary outcome. There is insufficient evidence to show whether paracetamol influenced the risk of febrile convulsions. In a meta analysis of two trials (n = 120), the proportion of children without fever by the second hour after treatment did not differ significantly between those given paracetamol and those sponged (Relative Risk 1.84; confidence interval 0.94 to 3.61, random effects model). The statistical test showed significant heterogeneity between the groups receiving paracetamol or physical methods. No severe adverse events were reported. The number of children with mild adverse events did not differ significantly between paracetamol and placebo, or paracetamol and physical methods, but numbers were small. REVIEWER'S CONCLUSIONS: Trial evidence that paracetamol has a superior antipyretic effect than placebo is inconclusive. There is limited evidence that there is no difference between the antipyretic effect of paracetamol and physical methods. Data on adverse events in these trials were limited. Establishing standard outcomes will help comparisons between studies and meta-analysis. PMID- 12076500 TI - Group-based parent-training programmes for improving emotional and behavioural adjustment in 0-3 year old children. AB - BACKGROUND: Mental health problems in children are common. Research suggests that parenting has an important role to play in helping children to become well adjusted adults, and that the first few months and years of a child's life are especially important in establishing patterns of emotional, cognitive and social functioning which will in turn influence the child's future development and in particular, their mental health. Parenting programmes may therefore have a role to play in improving the mental health of infants and toddlers. OBJECTIVES: The objectives of this review are as follows: a) To establish whether group-based parenting programmes are effective in improving the mental health of children less than three years of age b) To assess the role of parenting programmes in the primary prevention of mental health problems SEARCH STRATEGY: A range of biomedical and social science databases were searched including MEDLINE, EMBASE, CINAHL, PsychLIT, Sociofile, Social Science Citation Index, ASSIA, the Cochrane Library including SPECTR, CENTRAL, National Research Register (NRR) and ERIC. SELECTION CRITERIA: Only randomised controlled trials were included and studies had to include at least one standardised instrument measuring some aspect of infant mental health. DATA COLLECTION AND ANALYSIS: The treatment effect for each outcome in each study was standardised by dividing the mean difference in post intervention scores for the intervention and treatment group, by the pooled standard deviation, to obtain an effect size. The results for each outcome in each study have been presented with 95% confidence intervals. Where appropriate the results have been combined in a meta-analysis using a random effect model. MAIN RESULTS: A total of five studies were included in the review. The 5 included studies provided a total of 36 assessments of infant and toddler mental health including emotional and behavioural adjustment, and sleep patterns. All of the results apart from 6 showed positive findings favouring the intervention group. While some of findings were non-significant, most of the effect sizes were large and the wide confidence intervals crossing the zero that were obtained in a number of the studies, were very probably due to small sample sizes. There was sufficient data from four studies to combine the results in a meta-analysis. The results of the meta-analysis show a significant difference in children's emotional and behavioural adjustment favouring the intervention group. Overall, the limited follow-up data provide equivocal evidence concerning the maintenance of these effects over time. REVIEWER'S CONCLUSIONS: The results of this review suggest that parenting programmes can be effective in improving the mental health of infants and toddlers. There is, however, insufficient evidence to reach any firm conclusions regarding the role that such programmes might play in the primary prevention of mental health problems. Furthermore, there is insufficient evidence to know whether the short-term benefit of these programmes is maintained over time, and further research is required. PMID- 12076501 TI - Interventions for learning disabled sex offenders. AB - BACKGROUND: The management of sex offenders is a major public concern. Behavioural and pharmacological interventions have been used for many years and more recently cognitive behavioural based interventions have become popular around the world. Programmes designed for the general population have been modified for those sex offenders with learning disability, to address their cognitive deficits. The efficacy of these modified programmes is unclear. OBJECTIVES: To determine the efficacy of interventions with learning disabled sex offenders. SEARCH STRATEGY: The reviewers searched electronically EMBASE, PsycINFO, Medline, Cinahl, Cochrane Library, SPECTR, National Clearinghouse on Child Abuse and Neglect Information and National Criminal Justice Reference Service and Biological Abstracts. SELECTION CRITERIA: All randomised controlled trials comparing an intervention for learning disabled sex offenders to any other, or no intervention. DATA COLLECTION AND ANALYSIS: Data were independently extracted. MAIN RESULTS: No randomised controlled trial was identified. REVIEWER'S CONCLUSIONS: Using the methods described the reviewers found no randomised controlled trial evidence to guide the use of interventions for learning disabled sex offenders. Until the urgent need for randomised controlled trials is met, clinical practice will continue to be guided by either extrapolation of evidence from randomised controlled trials involving sex offenders without learning disability or non-randomised trial evidence of interventions for the learning disabled sex offender. PMID- 12076502 TI - Rofecoxib for the treatment of rheumatoid arthritis. AB - BACKGROUND: Rheumatoid arthritis (RA) is a systemic auto-immune disorder, in which the synovial lining of many joints and tendon sheaths are persistently inflamed. OBJECTIVES: To assess the efficacy and toxicity of rofecoxib for treating RA. SEARCH STRATEGY: We searched the following databases up to December 2000: MEDLINE, EMBASE, Cochrane Database of Systematic Reviews, Cochrane Controlled Trials Register, National Research Register, NHS Economic Evaluation Database, Health Technology Assessment database. The bibliographies of retrieved papers were scanned for additional references. The manufacturers of rofecoxib, MSD, were also approached by the UK National Institue for Clincal Excellence to submit additional evidence to inform it's appraisal on the use of cyclo-oxygenase inibitors for arthritis. SELECTION CRITERIA: Relevant studies were randomised controlled trials of parallel group design evaluating the efficacy and/or toxicity of rofecoxib in RA, both placebo based and comparative trials were eligible. Relevant outcome criteria had to be available to evaluate efficacy and/or toxicity, such as the OMERACT outcomes. DATA COLLECTION AND ANALYSIS: Data were abstracted independently by two reviewers and the results were compared for the degree of agreement. A validated tool (Jadad 1995) was used to score the quality of the randomised controlled trials. The planned analysis was to pool, where appropriate, continuous outcome measures using mean or standardized mean differences, and dichotomous outcome measures using relative risk ratios. MAIN RESULTS: Two randomised controlled trials evaluating the efficacy and toxicity of rofecoxib in RA were identified and met the criteria. One compared rofecoxib to placebo and the other compared rofecoxib to naproxen. The overall number of ACR 20 responders who had received 25mg (82/ 171 = 48%) or 50mg (86/161 = 53%) was statistically significantly more than those receiving placebo (58/168 = 35% ) (RR 1.39 CI: 1.07, 1.80 and RR 1.55 CI: 1.20, 1.99 respectively) with no statistically significant differences between the 25 and 50 mg doses. The safety profile of rofecoxib was similar to that of placebo. In the comparative trial, rofecoxib at a dosage of 50 mg/day demonstrated similar efficacy to naproxen at a dosage of 500 mg twice daily. However, the combined rate of clinically significant complicated gastro-intestinal events (GI) (perforations, ulcers, bleeds, or obstructions) was lower with rofecoxib than with naproxen (RR 0.46, 95% CI, 0.34 to 0.63) due to a reduction in the number of ulcers and bleeds. Patients taking rofecoxib had a greater risk of having a myocardial infarction (MI) than patients taking naproxen (RR 4.03, 95% CI, 2.86 to 5.68). REVIEWER'S CONCLUSIONS: In patients with RA, rofecoxib demonstrates a greater degree of efficacy than placebo, while having a comparable safety profile. Rofecoxib demonstrates a similar degree of efficacy as naproxen, but with a significantly lower rate of ulceration and gastrointestinal bleeding. Rofecoxib was associated with a greater risk for MI, but the exact significance and pathophysiology of this possible relationship is unclear. PMID- 12076503 TI - Non-steroidal anti-inflammatory drugs (NSAIDs) for treating lateral elbow pain in adults. AB - BACKGROUND: Lateral elbow pain, or tennis elbow, is a common condition causing pain in the elbow and forearm and lack of strength and function of the elbow and wrist. It is often treated with non-steroidal anti-inflammatory drugs (NSAIDs), either orally or by topical application. OBJECTIVES: To assess effectiveness of NSAIDs (oral or topical) in the treatment of adults with lateral elbow pain with respect to symptom (pain) reduction, improvement in function, grip strength and adverse effects. SEARCH STRATEGY: We searched the Musculoskeletal Review Group's trials register database, the Cochrane Clinical Trials Register (Cochrane Library issue 2, 2001), MEDLINE, CINAHL, EMBASE and SCISEARCH up to June 2001. SELECTION CRITERIA: Randomised and pseudo randomised trials in all languages of NSAIDs (oral or topical) compared to placebo or another intervention, or comparing two NSAIDs (oral or topical) to each other, in adults with lateral elbow pain (tennis elbow). Outcomes of interest were pain, function, disability and quality of life, strength, participant satisfaction with treatment and adverse effect. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied selection criteria and assessed study quality. MAIN RESULTS: Fourteen trials were included in the review. Few trials used intention to treat analysis, and the sample size of most was small. The median follow up was 2 weeks (range 1-12 weeks). There is evidence that topical NSAIDs are significantly more effective than placebo with respect to pain [weighted main difference= -1.88, (95% confidence intervals -2.54 to -1.21)] and participant satisfaction [relative risk 0.39, (95% confidence intervals 0.23 to 0.66)] in the short term, and this finding is robust against the possible bias introduced by the inclusion of unblinded trials and publication bias. The adverse effects reported were minor. Only two included trials assessed the effect of oral NSAID and these were not able to be pooled. There is some evidence for short term benefit with respect to pain and function from oral NSAIDs, but this benefit was not sustained. Significantly more gastrointestinal adverse effects were reported by those taking oral NSAIDs [relative risk = 3.17, (95% confidence intervals 1.35 to 7.41)]. In the short term there may be some advantage in steroid injection over oral NSAID [patient's perception of benefit relative risk = 3.06, (95% confidence intervals 1.55 to 6.06)], but this was not sustained in the longer term. REVIEWER'S CONCLUSIONS: There is some support for the use of topical NSAIDs to relieve lateral elbow pain at least in the short term. There remains insufficient evidence to recommend or discourage the use of oral NSAID, although it appears injection may be more effective than oral NSAID in the short term. A direct comparison between topical and oral NSAID has not been made and so no conclusions can be drawn regarding the best method of administration. PMID- 12076504 TI - Electrical stimulation for the treatment of rheumatoid arthritis. AB - BACKGROUND: Electrical stimulation is one of several rehabilitation interventions suggested for the management of rheumatoid arthritis (RA) to enhance muscle performance. OBJECTIVES: To assess the effectiveness of electrical stimulation for improving muscle strength and function in clients with RA. SEARCH STRATEGY: We searched MEDLINE, Embase, Healthstar, Sports Discus, CINAHL, the Cochrane Controlled Trials Register, the PEDro database, the specialized registry of the Cochrane musculoskeletal group and the Cochrane field of physical and related therapies up to January 2002 according to the sensitive search strategy for RCTs designed for the Cochrane Collaboration. The search was complemented with handsearching of the reference lists. Key experts in the area were contacted for further articles. SELECTION CRITERIA: All randomized controlled trials (RCTs) and controlled clinical trials (CCTs), case-control and cohort studies comparing ES against placebo or another active intervention in patients with RA were selected, according to an a priori protocol. No language restrictions were applied. DATA COLLECTION AND ANALYSIS: Two independent reviewers determined the studies to be included based on a priori inclusion criteria. Data were independently abstracted by the same two reviewers, and checked by a third reviewer using a pre-developed form. The same two reviewers, using a validated scale, independently assessed the methodological quality of the RCTs and CCTs. The data analysis was performed using Peto Odds ratios. MAIN RESULTS: Of the two relevant studies that were identified in the literature, only one RCT met the inclusion criteria. This RCT compared the effects of two electrostimulation (ES) protocols on hand function in general and on the performance of the first dorsal interosseous muscle in particular, in 15 patients with RA and secondary disuse atrophy of the first dorsal interosseous of the dominant hand. The results showed that ES had significant benefit when compared to a control no treatment group in terms of muscle strength and fatigue resistance of the first dorsal interosseous. Most favourable results were obtained by using a patterned stimulation derived from a fatigued motor unit of the first dorsal interosseous in a normal hand rather than a fixed 10 Hz stimulation frequency. Side effects of the ES application were not reported. REVIEWER'S CONCLUSIONS: ES was shown to have a clinically beneficial effect on grip strength and fatigue resistance for RA patients with muscle atrophy of the hand. However, these conclusions are limited by the low methodological quality of the trial included. More well-designed studies are therefore needed to provide further evidence of the benefits of ES in the management of RA. PMID- 12076505 TI - Patient education for adults with rheumatoid arthritis. AB - BACKGROUND: Because of the unpredictability people with arthritis face on a daily basis, patient education programmes have become an effective complement to traditional medical treatment giving people with arthritis the strategies and the tools necessary to make daily decisions to cope with the disease. OBJECTIVES: To assess the effectiveness of patient education interventions on health status in patients with rheumatoid arthritis. SEARCH STRATEGY: We searched MEDLINE, EMBASE and PsycINFO and the Cochrane Controlled Trials Register. A selection of review articles (see references) were examined to identify further relevant publications. There was no language restriction. SELECTION CRITERIA: Randomised controlled trials (RCT's) evaluating patient education interventions that included an instructional component and a non-intervention control group; pre- and post-test results available separately for RA, either in the publication or from the studies' authors; and study results presented in full, end-of-study report. MAIN RESULTS: Twenty-four studies with relevant data were included. We found significant effects of patient education at first follow-up for scores on disability, joint counts, patient global assessment and psychological status. Physician global assessment was not assessed in any of the included studies. The two separate dimensions of psychological status: anxiety and depression showed no significant effects, nor did the dimensions of pain and disease activity. At final follow up no significant effects of patient education were found. REVIEWER'S CONCLUSIONS: Patient education as provided in the studies reviewed here had moderate short-term effects on patient global assessment, and small short-term effects on disability, joint counts and psychological status. There were no long-term benefits. PMID- 12076506 TI - Arsenic toxicity and potential mechanisms of action. AB - Exposure to the metalloid arsenic is a daily occurrence because of its environmental pervasiveness. Arsenic, which is found in several different chemical forms and oxidation states, causes acute and chronic adverse health effects, including cancer. The metabolism of arsenic has an important role in its toxicity. The metabolism involves reduction to a trivalent state and oxidative methylation to a pentavalent state. The trivalent arsenicals, including those methylated, have more potent toxic properties than the pentavalent arsenicals. The exact mechanism of the action of arsenic is not known, but several hypotheses have been proposed. At a biochemical level, inorganic arsenic in the pentavalent state may replace phosphate in several reactions. In the trivalent state, inorganic and organic (methylated) arsenic may react with critical thiols in proteins and inhibit their activity. Regarding cancer, potential mechanisms include genotoxicity, altered DNA methylation, oxidative stress, altered cell proliferation, co-carcinogenesis, and tumor promotion. A better understanding of the mechanism(s) of action of arsenic will make a more confident determination of the risks associated with exposure to this chemical. PMID- 12076507 TI - A review of animal models for the study of arsenic carcinogenesis. AB - As inorganic arsenic is a proven human carcinogen, significant effort has been made in recent decades in an attempt to understand arsenic carcinogenesis using animal models, including rodents (rats and mice) and larger mammals such as beagles and monkeys. Transgenic animals were also used to test the carcinogenic effect of arsenicals, but until recently all models had failed to mimic satisfactorily the actual mechanism of arsenic carcinogenicity. However, within the past decade successful animal models have been developed using the most common strains of mice or rats. Thus dimethylarsinic acid (DMA), an organic arsenic compound which is the major metabolite of inorganic arsenicals in mammals, has been proven to be tumorigenic in such animals. Reports of successful cancer induction in animals by inorganic arsenic (arsenite and arsenate) have been rare, and most carcinogenetic studies have used organic arsenicals such as DMA combined with other tumor initiators. Although such experiments used high concentrations of arsenicals for the promotion of tumors, animal models using doses of arsenicals species closed to the exposure level of humans in endemic areas are obviously the most significant. Almost all researchers have used drinking water or food as the pathway for the development of animal model test systems in order to mimic chronic arsenic poisoning in humans; such pathways seem more likely to achieve desirable results. PMID- 12076508 TI - Effect of arsenic on transcription factor AP-1 and NF-kappaB DNA binding activity and related gene expression. AB - Both acute (24 h) and chronic (10-20 week) exposure of human fibroblast cells to low dose sodium arsenite (As(III)) significantly affects activating protein-1 (AP 1) and nuclear factor kappa B (NF-kappa B) DNA binding activity. Short-term treatment with 0.1-5 microM As(III) up-regulates expression of c-Fos and c-Jun and the redox regulators, thioredoxin (Trx) and Redox factor-1 (Ref-1) and activates both AP-1 and NF-kappa B binding. Chronic exposure to 0.1 or 0.5 microM As(III) decreased c-Jun, c-Fos and Ref-1 protein levels and AP-1 and NF-kappa B binding activity, but increased Trx expression. Short term exposure to phorbol 12 myristate 13-acetate (TPA), a phorbol ester tumour promoter, or hydrogen peroxide (H(2)O(2)) also activates AP-1 and NF-kappa B binding. However, pre-treatment with As(III) prevents this increase. These results suggest that As(III) may alter AP-1 and NF-kappa B activity, in part, by up-regulating Trx and Ref-1. The different effects of short- versus long-term As(III) treatment on acute-phase response to oxidative stress reflect changes in the expression of Ref-1, c-Fos and c-Jun, but not Trx. PMID- 12076509 TI - Plasmid DNA damage caused by methylated arsenicals, ascorbic acid and human liver ferritin. AB - Both dimethylarsinic acid (DMA(V)) and dimethylarsinous acid (DMA(III)) release iron from human liver ferritin (HLF) with or without the presence of ascorbic acid. With ascorbic acid the rate of iron release from HLF by DMA(V) was intermediate (3.37 nM/min, P<0.05) and by DMA(III) was much higher (16.3 nM/min, P<0.001). No pBR322 plasmid DNA damage was observed from in vitro exposure to arsenate (iAs(V)), arsenite (iAs(III)), monomethylarsonic acid (MMA(V)), monomethylarsonous acid (MMA(III)) or DMA(V) alone. DNA damage was observed following DMA(III) exposure; coexposure to DMA(III) and HLF caused more DNA damage; considerably higher amounts of DNA damage was caused by coexposure of DMA(III), HLF and ascorbic acid. Diethylenetriaminepentaacetic acid (an iron chelator), significantly inhibited DNA damage. Addition of catalase (which can increase Fe(2+) concentrations) further increased the plasmid DNA damage. Iron dependent DNA damage could be a mechanism of action of human arsenic carcinogenesis. PMID- 12076510 TI - Arsenic inhibits the repair of DNA damage induced by benzo(a)pyrene. AB - In order to study the effect of arsenic on DNA damage, Sprague-Dawley rats were dosed with sodium arsenite (10 mg/kg) with or without 800 microg of benzo(a)pyrene (BP) by intramammilary injection. The animals were sacrificed on day 1, 3, 5, 10 and 27 and the mammary gland tissues were collected for DNA adduct measurement using a (32)P post-labeling assay. Animals dosed with arsenic alone did not show any DNA adducts. DNA adduct levels in rats dosed with BP alone reached a maximum level by day 5, reducing to 13% of this level by day 27. Adduct levels in rats dosed with arsenic and BP also reached a maximum by day 5 but only 80% of the level observed in the BP group. However, 84% of this amount still remained by day 27. The First Nucleotide Change (FNC) technique was used for the screening of 115 samples of various tissues from mice that had been chronically exposed to sodium arsenate for over 2 years revealed that inorganic arsenic did not attack the two putative hotspots (codons 131 and 154) of the hOGG1 gene. These results support the hypothesis that arsenic exerts its biological activity through DNA repair inhibition. PMID- 12076511 TI - Epidemiologic evidence of diabetogenic effect of arsenic. AB - It is well documented that arsenic can lead to skin lesions, atherosclerotic diseases and cancers. The association between arsenic exposure and diabetes mellitus is a relatively new finding. Up to now, there are six epidemiologic reports linking diabetes mellitus with arsenic exposure from environmental and occupational sources. Two reports in Taiwan carried out in the blackfoot disease hyperendemic villages, one cross-sectional and one prospective follow-up of the same cohort, indicate that arsenic exposure from drinking artesian well water is associated with prevalence and incidence of diabetes mellitus in a dose responsive pattern. The observation of the relation between arsenic exposure and diabetes mellitus is further supported by studies carried out in Sweden and Bangladesh. In Sweden, case-control analyses of death records of copper smelters and glass workers revealed a trend of increasing diabetes mellitus with increasing arsenic exposure from inhalation. In Bangladesh, prevalence of diabetes mellitus among arsenic-exposed subjects with keratosis was about five times higher than unexposed subjects. Increasing trends of diabetes mellitus with indices of arsenic exposure in drinking water seems to be independent of the presence of skin lesions associated with arsenic exposure. Although these studies consistently show an association between arsenic exposure and diabetes mellitus, the weak study designs of cross-sectional or case-control, the use of glucosuria or diabetes death as diagnostic criteria and the lack of adjustment for possible confounders in some studies, are major limitations that may reduce the strength of the evidence. PMID- 12076513 TI - Urinary arsenic speciation in subjects with or without restriction from seafood dietary intake. AB - In order to understand whether ingestion of seafood affects the urinary arsenic metabolites. About 42 women and 36 men were recruited from the students, parents and teachers in Taipei Medical University and National Taiwan University. The study subjects were interviewed about dietary habits, cigarette smoking habits, drug and vitamin intake, and consumption of seafood. Urine samples were collected from study subjects before and after refraining from eating seafood for 3 days, respectively. The urine samples were frozen at -20 degrees C separated by high performance liquid chromatography (HPLC), and on line linked to hydride generator atomic absorption spectrometry (HGAAS) to quantify the levels of various species of inorganic arsenic and its metabolites. The levels of arsenite (AsIII), arsenate (AsV), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), total inorganic arsenic metabolites, inorganic arsenic percent, MMA percent and DMA percent were similar before and after refraining from eating seafood for 3 days. The frequencies of fish, shellfish and seaweed dietary intake were not significantly correlated with urinary arsenic species. PMID- 12076512 TI - The absorption and excretion of fluoride and arsenic in humans. AB - The absorption and excretion of fluoride and arsenic were measured in a group of healthy volunteers given drinking water with naturally high concentration of fluoride (F 2.3 mg/l)(,) or high concentration of arsenic (As 0.15 mg/l), or high concentrations of both fluoride and arsenic (F 2.25 mg/l, As 0.23 mg/l and F 4.05 mg/l, As 0.58 mg/l), respectively. The results indicated that, for arsenic, the absorption rate, the proportion of urinary excretion and the biological-half-life did not show statistically significant differences between drinking water containing high arsenic alone and drinking water containing different levels of high arsenic and fluoride. Excretion and retention of arsenic were positively correlated to the total arsenic intake. Similar results were observed for fluoride. This suggests that there are different metabolic processes for arsenic and fluoride in respect to absorption and excretion; and no joint action can be attributed by these two elements. PMID- 12076514 TI - HPLC measurement of harderoporphyrin in the harderian glands of rodents as a biomarker for sub-lethal or chronic arsenic exposure. AB - An improved HPLC method has been established for the measurement of harderoporphyrin (HP) in the harderian gland of rats and mice. Groups of female Wistar rats were given a single oral dose of sodium arsenite at 0, 0.5 or 5.0 mg As(III)/kg body weight, or a slurry of arsenic-contaminated soil at equivalent dose rates and the animals were sacrificed 96 h after dosing. A group of C57BL/6J female mice were chronically exposed to drinking water containing 500 microg As(V)/l of sodium arsenate ad libitum for over 2 years. Porphyrins were measured in the harderian glands of rats and mice. Our results suggest that HP and the alteration of the porphyrin profile in the harderian glands of rodents is a highly sensitive biomarker for both single sub-lethal and chronic arsenic exposure. PMID- 12076515 TI - Combined effects of anions on arsenic removal by iron hydroxides. AB - Batch experiments were conducted to investigate the combined effects of phosphate, silicate, and bicarbonate on the removal of arsenic from Bangladesh groundwater (BGW) and simulated groundwater by iron hydroxides. The apparent adsorption constants indicated that the affinity of the anions for iron hydroxide sites decreased in the following order arsenate>phosphate>arsenite>silicate>bicarbonate. Phosphate, silicate, and bicarbonate decreased the removal of As(III) even at relatively low concentrations and low surface site coverage. Phosphate (0-0.08 mM), silicate (0 0.8 mM), and bicarbonate (0-14 mM) in separate solutions had none to moderate effects on As(V) removal in a solution containing 6.7 mg/l Fe and 0.3 ppm As(V). In the presence of bicarbonate and silicate the adverse effect of phosphate on As(V) adsorption was magnified. The residual As(V) concentration after iron hydroxide treatment increased from less than 13 microg/l in separate bicarbonate (2.2 mM) and phosphate (0.062 mM) solutions to 110 microg/l in the solution containing both anions. The results suggested the combined effects of phosphate, silicate, and bicarbonate caused the high mobility of arsenic in Bangladesh water. PMID- 12076516 TI - Arsenic removal from water using advanced oxidation processes. AB - Consumption of groundwaters containing natural arsenic at several hundred microg/l (ppb) in countries such as Bangladesh has lead to the increased occurrence of many cancers particularly those of the skin and bladder, while concerns in the USA and Australia regarding the unknown health impact of drinking water containing tens of ppb of arsenic is leading to increasingly stringent maximum contaminant levels. The anaerobic conditions of these groundwaters result in the arsenic being present in its reduced form, hence the use of an oxidant is necessary if the arsenic is to be successfully removed by precipitation or ion exchange methods. Advance oxidation methods which utilise ultraviolet light and a photo absorber have been developed and patented, in which both iron salts and sulphite can be used as the photoabsorber. The former absorber has been developed for arsenic removal in rural areas of Bangladesh and the latter for groundwaters in countries such as the USA. PMID- 12076518 TI - Modulation of cytostatic efficacy of melphalan by glutathione: mechanisms and efficacy. PMID- 12076519 TI - Chemiluminescence associated with the oxidative metabolism of salicylic acid in rat liver microsomes. AB - Rat liver microsomal suspension (1 mg protein per ml) was incubated at 37 degrees C with 5 mM salicylic acid and 0.2 mM NADPH. The amounts of thiobarbituric acid reactive substances (TBARS) and 2,5-dihydroxybenzoic acid (2,5-DHB), an oxidative metabolite of salicylic acid increased with the incubation time. Simultaneously spontaneous chemiluminescence (CL) was found to be generated there. The addition of SKF-525A, an inhibitor of cytochrome P450 (P450), to the reaction mixture inhibited the CL generation together with the inhibition of the oxidative metabolism. The anti-oxidants and singlet oxygen scavengers like N,N diphenylphenylenediamine (DPPD) and histidine suppressed the CL generation. The addition of 1,4-diazabicyclo [2.2.2] octane (DABCO), a singlet oxygen quencher, to the reaction mixture generating CL enhanced CL transiently and then CL decreased markedly. Thus CL observed here may possibly originate from the singlet oxygen. The CL generation was suggested to be closely related with salicylic acid induced lipid peroxidation, and to be coupled with the oxidative metabolism mediated by P450 in rat liver microsomes. PMID- 12076520 TI - Expression of rat liver glutathione-S-transferase GSTA5 in cell lines provides increased resistance to alkylating agents and toxic aldehydes. AB - The glutathione-S-transferases (GST) are a major contributor to the eukaryotic cell's defences against chemical and oxidative stress. However, the role of individual GST isoenzymes in conferring resistance to xenobiotics has not been fully determined. We have examined the effect of the rat GSTA5 isoenzyme in the detoxication of alkylating agents and aldehydes by constructing a cell line in which it is stably expressed. The hamster fibroblast cell line V79 was transfected with a construct expressing GSTA5 from the CMV promoter. A stable clone (V79-GSTA5) was isolated after selecting for the neomycin phosphotransferase gene present on the introduced DNA. The cell line showed significantly increased levels of resistance towards the alkylating agents chorambucil and melphalan. Levels of resistance were 4-6-fold greater in V79 GSTA5 cells than in control cells. Increased levels of resistance were also observed towards the lipid peroxidation product acrolein (IC(50)=80 microM compared with 17 microM in control cells). The V79-GSTA5 cells also showed a 4 fold increase in resistance to trans, trans muconaldehyde (IC(50)=4 micro compared with l microM for control cells). GSTA5 did not protect against 4 hydroxynonenal, but it did provide greater levels of protection to hydrogen peroxide, with an IC(50) of 380 microM in V79-GSTA5 compared with 180 microM in control cells. In contrast, V79-GSTA5 cells were more sensitive to methyl glyoxal, suggesting that a methyl glyoxal-glutathione conjugate is more toxic that the parental compound. These data contribute towards the evaluation of the role of GSTA5 in the detoxication of these compounds. PMID- 12076522 TI - Kinetics of bromodichloromethane metabolism by cytochrome P450 isoenzymes in human liver microsomes. AB - The kinetic constants for the metabolism of bromodichloromethane (BDCM) by three cytochrome P450 (CYP) isoenzymes have been measured in human liver microsomes. The three CYP isoenzymes, CYP2E1, CYP1A2 and CYP3A4, have been identified previously as important in the metabolism of this compound. To measure the constants for each isoenzyme, enzyme-specific inhibitory antibodies were used to block the activities for two of the three isoenzymes. CYP2E1 was found to have the lowest K(m), 2.9 microM, and the highest catalytic activity, k(cat). The K(m) for the other isoenzymes, CYP1A2 and CYP3A4, were about 60 microM with lower values of k(cat). Apparent kinetic constants obtained from two microsomal samples that were not inhibited were consistent with these results. In addition, 11 human microsome samples characterized for 10 CYP activities were correlated with the metabolism of 9.7 microM BDCM by each sample; statistical analysis showed a correlation with CYP2E1 activity only. This result is consistent with the finding that CYP2E1 is the only isoenzyme with a K(m) lower than the BDCM concentration used. The kinetic constants obtained from the inhibited microsomes were compared to similar results from recombinant human isoenzyme preparations containing only one CYP isoenzyme. The results for CYP2E1 were very similar, while the results for CYP1A2 were somewhat less similar and there was a substantial divergence for CYP3A4 in the two systems. Possible reasons for these differences are differing levels of CYP reductase and/or differing makeup of the membrane lipid environment for the CYPs. Because of the low levels of BDCM exposure from drinking water, it appears likely that CYP2E1 will dominate hepatic CYP-mediated BDCM metabolism in humans. PMID- 12076521 TI - Quantitative evaluation of bromodichloromethane metabolism by recombinant rat and human cytochrome P450s. AB - We report quantitative estimates of the parameters for metabolism of bromodichloromethane (BDCM) by recombinant preparations of hepatic cytochrome P450s (CYPs) from rat and human. Earlier work identified CYP2E1, CYP2B1/2 and CYP1A2 as activating enzymes necessary for hepatotoxicity in rat. In order to extend an existing PBPK model for rat to include a capability for extrapolation to humans, it is necessary to evaluate quantitatively the principal metabolic pathways in both species. We have conducted in vitro experiments using recombinant preparations of the three rat CYP isoenzymes mentioned above and for CYP2C11 and CYP3A1 as well. Similar experiments have been performed with human recombinant isoenzymes for CYP2E1, CYP1A2, CYP2A6, CYP2B6, CYP2D6 and CYP3A4. Results indicate that the principal metabolizing enzymes in rat are those identified previously, CYP2E1, CYP2B1/2 and CYP1A2. CYP3A1 may also have some activity. In human, CYP2E1, CYP1A2 and CYP3A4 show substantial activity, and CYP2A6 also measurably metabolizes BDCM. In both species, CYP2E1 is the low K(m) isoenzyme, with K(m) approximately 27-fold lower than those for the isoenzymes with the next lowest K(m). In addition, the metabolic parameters, K(m) and k(cat), for rat and human CYP2E1 were nearly identical. The metabolic parameters for CYP1A2, the only other isoenzyme active in both species, were not similar across species. In addition, calculations based on the kinetic constants obtained are compared to results from two in vivo experiments to show that the in vitro kinetic data is relevant to in vivo exposures. We conclude that although several CYPs metabolize BDCM, at low concentration/exposure, BDCM metabolism is dominated by CYP2E1 in both rat and human, but that other isoenzymes can be important at higher concentrations. We further conclude that the kinetic data are consistent with existing in vivo results. PMID- 12076523 TI - Thiol protecting agents and antioxidants inhibit the mitochondrial permeability transition promoted by etoposide: implications in the prevention of etoposide induced apoptosis. AB - Etoposide (VP-16) is known to promote cell apoptosis either in cancer or in normal cells as a side effect. This fact is preceded by the induction of several mitochondrial events, including increase in Bax/Bcl-2 ratio followed by cytochrome c release and consequent activation of caspase-9 and -3, reduction of ATP levels, depolarization of membrane potential (DeltaPsi) and rupture of the outer membrane. These events are apoptotic factors essentially associated with the induction of the mitochondrial permeability transition (MPT). VP-16 has been shown to stimulate the Ca2+-dependent MPT induction similarly to prooxidants and to promote apoptosis by oxidative stress mechanisms, which is prevented by glutathione (GSH) and N-acetylcysteine (NAC). Therefore, the aim of this work was to study the effects of antioxidants and thiol protecting agents on MPT promoted by VP-16, attempting to identify the underlying mechanisms on VP-16-induced apoptosis. The increased sensitivity of isolated mitochondria to Ca2+-induced swelling, Ca2+ release, depolarization of DeltaPsi and uncoupling of respiration promoted by VP-16, which are prevented by cyclosporine A proving that VP-16 induces the MPT, are also efficiently prevented by ascorbate, the primary reductant of the phenoxyl radicals produced by VP-16. The thiol reagents GSH, dithiothreitol and N-ethylmaleimide, which have been reported to prevent the MPT induction, also protect this event promoted by VP-16. The inhibition of the VP-16 induced MPT by antioxidants agrees with the prevention of etoposide-induced apoptosis by GSH and NAC and suggests the generation of oxidant species as a potential mechanism underlying the MPT that may trigger the release of mitochondrial apoptogenic factors responsible for apoptotic cascade activation. PMID- 12076524 TI - The antimicrobial properties of milkfat after partial hydrolysis by calf pregastric lipase. AB - Studies on the kinetic characteristics of calf pregastric lipase (EC 3.1.1.3) have shown that it preferentially releases short chain fatty acids (SCFAs) from bovine milkfat. The released fatty acids form mixed micelle structures. The aim of this investigation has been to test whether hydrolysed milkfat is antimicrobial, and how the state of the emulsion alters the bactericidal or bacteriostatic effects. Partial hydrolysis of milkfat by pregastric lipase was carried out in two types of emulsion systems, containing either Triton X-100 or casein/lecithin, plus milkfat in citrate/phosphate buffer (pH 5.0-6.0). The concentrations and compositions of fatty acids were determined by gas chromatography. The minimum percentages of hydrolysed milkfat which affected growth and survival of selected Gram-positive and Gram-negative bacteria were measured. The bacterial experiments were repeated using pure fatty acids at similar concentrations. Lauric acid (C12:0) was found to be the most potent bactericidal fatty acid against Enterococcae (Gram-positive), and caprylic acid (C8:0) was the most potent against coliforms (Gram-negative). Use of Triton X-100 for milkfat emulsification provided a more compatible medium for studying bacterial growth in the hydrolysed milkfat than did use of casein/lecithin. The results also show that the antimicrobial effects of individual fatty acids released from hydrolysed milkfat were at least additive and suggest that hydrolysis of milkfat may be a significant factor in controlling growth of organisms imbibed with food in pre-weaned animals. The amount of pregastric catalyzed triglyceride hydrolysis in the digestive tract is sufficient to produce an antibacterial concentration of fatty acids and monoglycerides. PMID- 12076526 TI - ML -- a conserved domain involved in innate immunity and lipid metabolism. AB - ML (MD-2-related lipid-recognition) is a novel domain identified in MD-1, MD-2, GM2A, Npc2 and multiple proteins of unknown function in plants, animals and fungi. These single-domain proteins were predicted to form a beta-rich fold containing multiple strands, and to mediate diverse biological functions through interacting with specific lipids. PMID- 12076525 TI - Can boron get bacteria talking? PMID- 12076528 TI - Of forks and ends. PMID- 12076527 TI - Viral RNA-polymerases -- a predicted 2'-O-ribose methyltransferase domain shared by all Mononegavirales. AB - The Mononegavirales virus group comprises several major human pathogens, including measles, rabies and Ebola viruses. This article reports a computational analysis of the C-terminal region of RNA-dependent RNA-polymerases from Mononegavirales. Using a combination of sequence similarity and threading analysis, a 2'-O-ribose methyltransferase domain was identified that is involved in the capping of viral mRNAs. PMID- 12076529 TI - Eyeing up tryptophanyl-tRNA synthetase. PMID- 12076530 TI - Starvation amidst plenty: PDKs and diabetes mellitus. PMID- 12076532 TI - Transcriptional memory during cell division. PMID- 12076534 TI - Membrane proteins: shaping up. AB - Over recent years, much progress has been made in the identification and characterization of factors involved in the biosynthesis of integral membrane proteins of the helix-bundle type. In addition, our knowledge of membrane protein structure and the forces stabilizing helix-helix interactions in a lipid environment is expanding rapidly. However, it is still not clear how a membrane protein folds into its final form in vivo, nor what constraints there are on the folded structure that results from the mechanistic details of translocon-mediated assembly rather than simply from the thermodynamics of protein-lipid interactions. PMID- 12076535 TI - The SOCS box: a tale of destruction and degradation. AB - Although initially identified in the suppressor of cytokine signaling (SOCS) family of proteins, the C-terminal SOCS box has now been identified in more than 40 proteins in nine different families. Growing evidence suggests that the SOCS box, similar to the F-box, acts as a bridge between specific substrate-binding domains and the more generic proteins that comprise a large family of E3 ubiquitin protein ligases. In this way, SOCS proteins regulate protein turnover by targeting proteins for polyubiquitination and, therefore, for proteasome mediated degradation. PMID- 12076536 TI - Quaternary structure and function of transport proteins. AB - Membranes are important sites for the regulation of metabolic functions because they contain transport molecules, which often catalyze the first step in a pathway, and signal-transduction components, which allow the cell to communicate with the environment. Given the catalytic importance of transport proteins and their role in membrane stability, it is possible that oligomerization is used to regulate their function. This review evaluates knowledge of the functions that are associated with the oligomeric organization of secondary transport proteins, which are a major class of solute-translocation systems in all living species. PMID- 12076538 TI - Snap-shots of clathrin-mediated endocytosis. AB - Clathrin-mediated endocytosis is one of the major entry routes into a eukaryotic cell. It is driven by protein components that aid the selection of cargo and provide the mechanical force needed to both deform the plasma membrane and detach a vesicle. Clathrin-coated vesicles were first observed by electron microscopy in the early 1960s. In subsequent years, many of the characteristic intermediates generated during vesicle formation have been trapped and observed. A variety of electron microscopy techniques, from the analysis of sections through cells to the study of endocytic intermediates formed in vitro, have led to the proposition of a sequence of events and of roles for different proteins during vesicle formation. In this article, these techniques and the insights gained are reviewed, and their role in providing snap-shots of the stages of endocytosis in atomic detail is discussed. PMID- 12076537 TI - P450 BM3: the very model of a modern flavocytochrome. AB - Flavocytochrome P450 BM3 is a bacterial P450 system in which a fatty acid hydroxylase P450 is fused to a mammalian-like diflavin NADPH-P450 reductase in a single polypeptide. The enzyme is soluble (unlike mammalian P450 redox systems) and its fusion arrangement affords it the highest catalytic activity of any P450 mono-oxygenase. This article discusses the fundamental properties of P450 BM3 and how progress with this model P450 has affected our comprehension of P450 systems in general. PMID- 12076539 TI - beta-Actinin, Cap Z, connectin and titin: what's in a name? PMID- 12076540 TI - Resources for full-length cDNAs. PMID- 12076541 TI - Lost in translation. PMID- 12076544 TI - Getting a handle on obesity. PMID- 12076545 TI - Need for attention to mental health of young offenders. PMID- 12076546 TI - Is there a future for telemedicine? PMID- 12076547 TI - Coagulation signals for intact blood vessels. PMID- 12076548 TI - Bridges to Iran. PMID- 12076549 TI - HIV/AIDS in Africa, 2002. PMID- 12076550 TI - Joint teleconsultations (virtual outreach) versus standard outpatient appointments for patients referred by their general practitioner for a specialist opinion: a randomised trial. AB - BACKGROUND: The current model of general practitioner referral of patients to hospital specialists in the UK is sometimes associated with unnecessary duplication of investigations and treatments. We aimed to compare joint teleconsultations between general practitioners, specialists, and patients (virtual outreach) with standard outpatient referral. METHODS: Virtual outreach services were established in London and Shrewsbury. The general practitioners referred 3170 patients, of whom 2094 consented to participate in the study and were eligible for inclusion. 1051 patients were randomly assigned virtual outreach, and 1043 standard outpatient appointments. We followed up the patients for 6 months after their index consultation. The primary outcome measure was the offer of a follow-up outpatient appointment. Analysis was by intention to treat. FINDINGS: More patients in the virtual outreach group than the standard group were offered a follow-up appointment (502 [52%] vs 400 [41%], odds ratio 1.52 [95% CI 1.27-1.82], p<0.0001). Significant differences in effects were observed between the two sites (p=0.009) and across different specialties (p<0.0001). Virtual outreach increased the offers of follow-up appointments more in Shrewsbury than in London, and more in ear, nose, and throat surgery and orthopaedics than in the other specialties. Fewer tests and investigations were ordered in the virtual outreach group by an average of 0.79 per patient (0.37 1.21, p=0.0002). Patients' satisfaction (analysed per protocol) was greater after a virtual outreach consultation than after a standard outpatient consultation (mean difference 0.33 scale points [95% CI 0.23-0.43], p<0.0001), with no heterogeneity between specialties or sites. INTERPRETATION: The trial showed that allocation of patients to virtual outreach consultations is variably associated with increased offers of follow-up appointments according to site and specialty, but leads to significant increases in patients' satisfaction and substantial reductions in tests and investigations. Efficient operation of such services will require appropriate selection of patients, significant service reorganisation, and provision of logistical support. PMID- 12076551 TI - Effects of fruit and vegetable consumption on plasma antioxidant concentrations and blood pressure: a randomised controlled trial. AB - BACKGROUND: High dietary intakes of fruit and vegetables are associated with reduced risks of cancer and cardiovascular disease. Short-term intensive dietary interventions in selected populations increase fruit and vegetable intake, raise plasma antioxidant concentrations, and lower blood pressure, but long-term effects of interventions in the general population are not certain. We assessed the effect of an intervention to increase fruit and vegetable consumption on plasma concentrations of antioxidant vitamins, daily fruit and vegetable intake, and blood pressure. METHODS: We undertook a 6-month, randomised, controlled trial of a brief negotiation method to encourage an increase in consumption of fruit and vegetables to at least five daily portions. We included 690 healthy participants aged 25-64 years recruited from a primary-care health centre. FINDINGS: Plasma concentrations of alpha-carotene, beta-carotene, lutein, beta cryptoxanthin, and ascorbic acid increased by more in the intervention group than in controls (significance of between-group differences ranged from p=0.032 to 0.0002). Groups did not differ for changes in lycopene, retinol, alpha tocopherol, gamma-tocopherol, or total cholesterol concentrations. Self-reported fruit and vegetable intake increased by a mean 1.4 (SD 1.7) portions in the intervention group and by 0.1 (1.3) portion in the control group (between-group difference=1.4, 95% CI 1.2-1.6; p<0.0001). Systolic blood pressure fell more in the intervention group than in controls (difference=4.0 mm Hg, 2.0-6.0; p<0.0001), as did diastolic blood pressure (1.5 mm Hg, 0.2-2.7; p=0.02). INTERPRETATION: The effects of the intervention on fruit and vegetable consumption, plasma antioxidants, and blood pressure would be expected to reduce cardiovascular disease in the general population. PMID- 12076552 TI - Mental health needs of boys in secure care for serious or persistent offending: a prospective, longitudinal study. AB - BACKGROUND: The mental health needs of children and adolescents in secure care are a matter of concern, but little systematic research has been done. Our aim was to assess the mental health, social, and educational needs of these young people in a prospective, longitudinal study. METHODS: We enrolled 97 boys aged 12 17 years who had been admitted to secure care. We assessed their needs (n=97) at the time of admission and 3 months later (n=90) with standardised interviews and psychometric tests. RESULTS: 26 (27%) boys had an intelligence quotient (IQ) of less than 70. The need for psychiatric help was high on admission to a secure unit, with the most frequent disorders being depression and anxiety. There were high rates of aggression, substance misuse, self harm, and social, family, and educational problems, and associated needs. The mean number of needs was 8.5 (SD 2.9) on admission and 2.9 (SD 2.4) after 3 months (mean difference 5.6, 95% CI 5.0-6.3). Areas in which needs were mostly met included education, substance misuse, self care, and diet. Areas where the frequency of need fell substantially, but remained high, were social and family problems, and aggressive behaviours. Psychological needs persisted, with new onsets of depression, anxiety problems, and post-traumatic-stress symptoms shortly after admission. The most frequently required interventions were psychological assessment and cognitive behavioural work. INTERPRETATION: Boys in secure care have many needs and a high rate of psychiatric morbidity. During the admission period, secure care units address some domains of need, but others remain unchanged or get worse. Psychological and psychiatric provision in secure units need to be improved. PMID- 12076553 TI - Feasibility and cost-effectiveness of standardised second-line drug treatment for chronic tuberculosis patients: a national cohort study in Peru. AB - BACKGROUND: There are no data on the feasibility and cost-effectiveness of using second-line drugs to treat patients with chronic tuberculosis, many of whom are infected with multidrug resistant (MDR) strains of Mycobacterium tuberculosis, in low or middle-income countries. METHODS: A national programme to treat chronic tuberculosis patients with a directly observed standardised 18-month daily regimen, consisting of kanamycin (3 months only), ciprofloxacin, ethionamide, pyrazinamide, and ethambutol, was established in Peru in 1997. Compliance and treatment outcomes were analysed for the cohort started on treatment between October, 1997, and March, 1999. Total and average costs were assessed. Cost effectiveness was estimated as the cost per DALY gained. FINDINGS: 466 patients were enrolled; 344 were tested for drug susceptibility and 298 (87%) had MDR tuberculosis. 225 patients (48%) were cured, 57 (12%) died, 131 (28%) did not respond to treatment, and 53 (11%) defaulted. Of the 413 (89%) patients who complied with treatment, 225 (55%) were cured. Among MDR patients, resistance to five or more drugs was significantly associated with an unfavourable outcome (death, non-response to treatment, or default; odds ratio 3.37, 95% CI 1.32-8.60; p=0.01). The programme cost US $0.6 million per year, 8% of the National Tuberculosis Programme budget, and US $2381 per patient for those who completed treatment. The mean cost per DALY gained was $211 ($165 at drug prices projected for 2002). INTERPRETATION: Treating chronic tuberculosis patients with high levels of MDR with second-line drugs can be feasible and cost-effective in middle income countries, provided a strong tuberculosis control programme is in place. PMID- 12076554 TI - Congestive heart failure and expression of myocardial urotensin II. AB - BACKGROUND: Human urotensin II has several cardiovascular actions, including potent vasoactive, and cardiac inotropic and hypertropic properties. Our aim was to ascertain degree of expression of urotensin II and its receptor GPR14 (now known as UT receptor) in the myocardium of patients with congestive heart failure (CHF). METHODS: We obtained specimens of myocardium from the hearts of 19 patients with end-stage CHF (12 ischaemic heart disease, seven dilated cardiomyopathy), five patients with early-stage CHF, and eight healthy controls. We used immunohistochemistry, in-situ hybridisation, reverse transcriptase-PCR (RT-PCR), and fluorescein isothiocyanate (FITC)-conjugated urotensin II to ascertain degree of myocardial expression of urotensin II and binding urotensin receptor. FINDINGS: Our results showed strong expression of urotensin II in the cardiomyocytes, and to a lesser extent in the vascular smooth muscle cells, endothelial cells, and inflammatory cells of patients with end-stage CHF. There was significantly less urotensin II expression in the myocardium of patients with early-stage CHF (p<0.0001). Also, there was little to no urotensin II expression in the myocardium of healthy controls. Myocardial expression of urotensin II correlated significantly with left ventricular end-diastolic dimension (p=0.0092), and inversely with ejection fraction (p=0.0002). RT-PCR showed increased concentrations of urotensin II and presence of urotensin receptor mRNA in the myocardium of patients with CHF. The confocal microscopy results showed a significant increase in the binding sites for urotensin in the myocardium of patients with end-stage CHF (p<0.0001). INTERPRETATION: Our findings suggest a possible role for urotensin II in the cardiac dysfunction and remodelling characteristic of CHF. PMID- 12076555 TI - Rejection, recurrence, or non-adherence? PMID- 12076556 TI - Multiple pulmonary arteriovenous malformations. PMID- 12076557 TI - White dysplastic melanocytic naevi. AB - We describe three cases of a new variant of dysplastic melanocytic naevi (DMN), which is characterised by complete lack of pigmentation. Clinically, white DMN were non-pigmented white to pale-red macules with accentuated skin markings, and had a silvery, shiny appearance under tangential light. Histology showed characteristic features of DMN--ie, non-typical melanocytes arranged as solitary units and nests at the dermoepidermal junction and in the papillary dermis, and mild stromal changes. Association with malignant melanoma in all three patients emphasises the importance of this morphological type of DMN as a possible precursor to, or marker of, malignant melanoma. PMID- 12076558 TI - Mapping of the wet/dry earwax locus to the pericentromeric region of chromosome 16. AB - Human earwax is a one-gene trait comprising two phenotypically distinct forms- wet and dry. This trait is attributed to secretory products of the ceruminous apocrine glands, and frequencies of phenotypes vary between ethnic groups. We did linkage analysis of eight Japanese families segregating earwax dimorphism. We assigned the earwax locus within a approximately 7.42-cM region between the loci D16S3093 and D16S3080 on chromosome 16p11.2-16q12.1, with a maximum two-point LOD score of 11.15 (theta;=0.00) at the locus D16S3044. Identification of the earwax locus could contribute to further anthropogenetic studies and physiological and pathological understanding of the apocrine-gland development. PMID- 12076559 TI - An epidemic Burkholderia cepacia complex strain identified in soil. AB - Life threatening infection with species of the Burkholderia cepacia complex frequently occurs as a result of cross infection among individuals with cystic fibrosis. Stringent infection control measures have decreased but not eliminated such infection in this vulnerable population, implying that non-patient reservoirs contribute to ongoing acquisition. However, strains common to both the natural environment and patients with cystic fibrosis have not yet been described. By use of various genotyping methods, we have identified from agricultural soil the B cepacia genomovar III strain that is most frequently recovered from cystic fibrosis patients in the mid-Atlantic region of the USA. This finding indicates that human pathogenic strains are not necessarily distinct from environmental strains, and might help explain ongoing human acquisition despite strict infection control measures. PMID- 12076560 TI - Breastfeeding and lowering the risk of childhood obesity. AB - Breastfeeding might confer protection against obesity later in life, but the evidence is inconclusive. We tested the hypothesis that breastfeeding is associated with a reduced risk of obesity in a population-based sample of 32200 Scottish children studied at age 39-42 months in 1998 and 1999. Obesity was defined as body-mass index (BMI) at the 95th and 98th percentiles or higher. The prevalence of obesity was significantly lower in breastfed children, and the association persisted after adjustment for socioeconomic status, birthweight, and sex. The adjusted odds ratio for obesity (BMI > or = 98th percentile) was 0.70 (95% CI 0.61-0.80). Our results suggest that breastfeeding is associated with a reduction in childhood obesity risk. PMID- 12076561 TI - Experts see progress in fight against tuberculosis. PMID- 12076564 TI - Are dyskinesias a problem after neural transplantation for Parkinson's disease? PMID- 12076562 TI - Researchers solve the mystery of nitroglycerin's mechanism of action. PMID- 12076565 TI - RNAi could hold promise in the treatment of HIV. PMID- 12076566 TI - A therapeutic future for CD44 in inflammation? PMID- 12076567 TI - UNICEF asks donors to give poorer nations longer-term commitment. PMID- 12076568 TI - Canadians are fighting fit, says government report. PMID- 12076569 TI - South Asia's thirsty cities search for solutions. PMID- 12076570 TI - The spread and effect of HIV-1 infection in sub-Saharan Africa. AB - Africa is the continent most severely affected by the global HIV-1 epidemic, with east and southern Africa in general more severely affected than west and central Africa. Differences in the spread of the epidemic can be accounted for by a complex interplay of sexual behaviour and biological factors that affect the probability of HIV-1 transmission per sex act. Sexual behaviour patterns are determined by cultural and socioeconomic contexts. In sub-Saharan Africa, some traditions and socioeconomic developments have contributed to the extensive spread of HIV-1 infection, including the subordinate position of women, impoverishment and decline of social services, rapid urbanisation and modernisation, and wars and conflicts. Populations in many parts of Africa are becoming trapped in a vicious circle as the HIV-1 epidemic leads to high mortality rates in young and economically productive age groups, and thus leads to further impoverishment. Interventions to control HIV-1 should not only target individuals, but also aim to change those aspects of cultural and socioeconomic context that increase the vulnerability to HIV-1 of people and communities. PMID- 12076571 TI - Treatment of postmenopausal osteoporosis. AB - The aim of treatment of postmenopausal osteoporosis is to reduce the frequency of vertebral and non-vertebral fractures (especially at the hip), which are responsible for morbidity associated with the disease. Results of large placebo controlled trials have shown that alendronate, raloxifene, risedronate, the 1-34 fragment of parathyroid hormone, and nasal calcitonin, greatly reduce the risk of vertebral fractures. Furthermore, a large reduction of non-vertebral fractures has been shown for alendronate, risedronate, and the 1-34 fragment of parathyroid hormone. Calcium and vitamin D supplementation is not sufficient to treat individuals with osteoporosis but is useful, especially in elderly women in care homes. Hormone replacement therapy remains a valuable option for the prevention of osteoporosis in early postmenopausal women. Choice of treatment depends on age, the presence or absence of prevalent fractures, especially at the spine, and the degree of bone mineral density measured at the spine and hip. Non pharmacological interventions include adequate calcium intake and diet, selected exercise programmes, reduction of other risk factors for osteoporotic fractures, and reduction of the risk of falls in elderly individuals. PMID- 12076572 TI - A developing country perspective. PMID- 12076573 TI - The soft science of German cardiology. PMID- 12076574 TI - Time to right the wrongs: improving basic health care in Nigeria. AB - Nigeria, once heralded as the beacon of Africa, has fallen somewhat short of this potential. Years of kleptocratic repressive dictators and military rule, coupled with widespread corruption, have resulted in large-scale neglect and deterioration of public services. Nowhere is this more apparent than within the health sector. Government-run health-care services barely function: half the population are unvaccinated for routine diseases, and a burgeoning epidemic of HIV/AIDS, only now being adequately addressed, leaves 3.5 million already infected and without access to the most basic of care. A poorly structured health service that relies on vertical programmes for HIV, tuberculosis, and malaria, means that coordination is chaotic, and already scant resources fail to reach the lower levels in which they are needed most. I visited Nigeria in October, 2001, with Medecins Sans Frontieres, a humanitarian aid organisation that has been working in Nigeria since 1996. I witnessed the poor level of health care in Nigeria for myself--a country that is more than capable of providing effective services--and concluded that, even now, political priorities are being put ahead of the population's basic needs. The challenges to the new civilian government are monumental, and it is yet to show any solid commitment to improving the health of Africa's biggest nation. PMID- 12076575 TI - Not so NICE for CML. PMID- 12076576 TI - Science, philosophy, religion, and use of embryonic stem cells. PMID- 12076578 TI - Long-term increase in bone mass through high calcium intake before puberty. PMID- 12076580 TI - Effects of folic acid. PMID- 12076581 TI - Effects of folic acid. PMID- 12076582 TI - Effects of folic acid. PMID- 12076583 TI - HIV-1 infection and intraepithelial neoplasia of lower genital tract. PMID- 12076584 TI - Progressive lateral ventricular enlargement as a clue to Alzheimer's disease. PMID- 12076585 TI - Family planning in Bangladesh. PMID- 12076586 TI - Physicians' charter and the new professionalism. PMID- 12076587 TI - Physicians' charter and the new professionalism. PMID- 12076588 TI - Physicians' charter and the new professionalism. PMID- 12076592 TI - Finding a voice. PMID- 12076589 TI - A complicated labour in 404 AD. PMID- 12076623 TI - Redescription of Neospora caninum and its differentiation from related coccidia. AB - Neospora caninum is a protozoan parasite of animals, which before 1984 was misidentified as Toxoplasma gondii. Infection by this parasite is a major cause of abortion in cattle and causes paralysis in dogs. Since the original description of N. caninum in 1988, considerable progress has been made in the understanding of its life cycle, biology, genetics and diagnosis. In this article, the authors redescribe the parasite, distinguish it from related coccidia, and provide accession numbers to its type specimens deposited in museums. PMID- 12076624 TI - Lymphatic filariasis: new insights into an old disease. AB - Lymphatic filariasis has afflicted people in the tropical areas of the world for thousands of years but even up to comparatively recent times it has been poorly understood and its importance under recognised. In the last 2 decades or so there has been a flurry of activity in filariasis research, which has provided new insights into the global problem of filariasis, the pathogenesis of filarial disease, diagnosis and control. PMID- 12076625 TI - Characterisation of hexokinase in Toxoplasma gondii tachyzoites. AB - We have cloned the hexokinase [E.C. 2.7.1.1] gene of Toxoplasma gondii tachyzoite and obtained an active recombinant enzyme with a calculated molecular mass of 51,465Da and an isoelectric point of 5.82. Southern blot analysis indicated that the hexokinase gene existed as a single copy in the tachyzoites of T. gondii. The sequence of T. gondii hexokinase exhibited the highest identity (44%) to that of Plasmodium falciparum hexokinase and lower identity of less than 35% to those of hexokinases from other organisms. The specific activity of the homogeneously purified recombinant enzyme was 4.04 micromol/mg protein/min at 37 degrees C under optimal conditions. The enzyme could use glucose, fructose, and mannose as substrates, though it preferred glucose. Adenosine triphosphate was exclusively the most effective phosphorus donor, and pyrophosphate did not serve as a substrate. K(m) values for glucose and adenosine triphosphate were 8.0+/-0.8 microM and 1.05+/-0.25mM, respectively. No allosteric effect of substrates was observed, and the products, glucose 6-phosphate and adenosine diphosphate, had no inhibitory effect on T. gondii hexokinase activity. Other phosphorylated hexoses, fructose 6-phosphate, trehalose 6-phosphate which is an inhibitor of yeast hexokinase, and pyrophosphate, also did not affect T. gondii hexokinase activity. Native hexokinase activity was recovered in both the cytosol and membrane fractions of the whole lysate of T. gondii tachyzoites. This result suggests that T. gondii hexokinase weakly associates with the membrane or particulate fraction of the tachyzoite cell. PMID- 12076626 TI - Pyridinylimidazole p38 mitogen-activated protein kinase inhibitors block intracellular Toxoplasma gondii replication. AB - Toxoplasma gondii is a medically important, obligate intracellular parasite. Little is known regarding factors that regulate its replication within cells. Such knowledge would further understanding of T. gondii pathogenesis, and might lead to novel therapeutic strategies. Mitogen-activated protein kinases (MAPKs) govern diverse cellular processes including proliferation and differentiation. We now show that treatment of T. gondii-infected cells with SB203580 or SB202190, substituted pyridinylimidazoles that are potent inhibitors of human p38 MAPK, inhibits intracellular T. gondii replication. Several independent experimental approaches suggest that the anti-proliferative effects of pyridinylimidazoles depend on direct action on tachyzoites, not the host cell: (i) selective inhibition of host p38 MAPK using recombinant adenoviruses had little effect on tachyzoite replication, (ii) pyridinylimidazole-treated tachyzoites developed abnormal morphology suggesting defective parasite division, and (iii) pyridinylimidazole-resistant mutant tachyzoites were developed through culture in progressively higher drug concentrations. We hypothesise that pyridinylimidazoles target a human p38 MAPK homologue in tachyzoites that regulates their replication. Phylogenetic data suggest that T. gondii likely encodes a p38 MAPK homologue, but such a homologue is absent from the incomplete Toxoplasma genomic data base. As all eukaryotic pathogens, including agents of malaria, leishmaniasis and trypanosomiasis encode endogenous MAPKs, drugs inhibiting endogenous MAPK activation may represent a novel, potentially broadly-acting class of anti-parasitic agents. Pyridinylimidazoles also represent tools to elucidate factors governing intracellular tachyzoite replication. PMID- 12076627 TI - An empirical quantitative framework for the seasonal population dynamics of the tick Ixodes ricinus. AB - The wide geographic and climatic range of the tick Ixodes ricinus, and the consequent marked variation in its seasonal population dynamics, have a direct impact on the transmission dynamics of the many pathogens vectored by this tick species. We use long-term observations on the seasonal abundance and fat contents (a marker of physiological ageing) of ticks, and contemporaneous microclimate at three field sites in the UK, to establish a simple quantitative framework for the phenology (i.e. seasonal cycle of development) of I. ricinus as a foundation for a generic population model. An hour-degree tick inter-stadial development model, driven by soil temperature and including diapause, predicts the recruitment (i.e. emergence from the previous stage) of a single cohort of each stage of ticks each year in the autumn. The timing of predicted emergence coincides exactly with the new appearance of high-fat nymphs and adults in the autumn. Thereafter, fat contents declined steadily until unfed ticks with very low energy reserves disappeared from the questing population within about 1 year from their recruitment. Very few newly emerged ticks were counted on the vegetation in the autumn, but they appeared in increasing numbers through the following spring. Larger ticks became active and subsequently left the questing population before smaller ones. Questing tick population dynamics are determined by seasonal patterns of tick behaviour, host-contact rates and mortality rates, superimposed on a basal phenology that is much less complex than has hitherto been portrayed. PMID- 12076628 TI - The population dynamics of Ostertagia gruehneri in reindeer: a model for the seasonal and intensity dependent variation in nematode fecundity. AB - The gastrointestinal nematode Ostertagia gruehneri is a parasite of reindeer that can have a significant impact on host population dynamics. To gain a better understanding of the population dynamics of O. gruehneri, we parameterise a model for its fecundity that describes the observed seasonal and intensity dependent pattern of faecal egg counts well. The faecal egg count model is combined with a model for the seasonal faecal production rate of Svalbard reindeer to obtain quantitative estimates of the fecundity of O. gruehneri. The model is used to evaluate the relative contribution to pasture contamination of variation in the abundance of O. gruehneri and variation in reindeer densities. It is concluded that due to the intensity dependence in nematode fecundity, variation in reindeer population densities is likely to be the most important of these factors for pasture contamination. PMID- 12076629 TI - Coastal freshwater runoff is a risk factor for Toxoplasma gondii infection of southern sea otters (Enhydra lutris nereis). AB - The association among anthropogenic environmental disturbance, pathogen pollution and the emergence of infectious diseases in wildlife has been postulated, but not always well supported by epidemiologic data. Specific evidence of coastal contamination of the marine ecosystem with the zoonotic protozoan parasite, Toxoplasma gondii, and extensive infection of southern sea otters (Enhydra lutris nereis) along the California coast was documented by this study. To investigate the extent of exposure and factors contributing to the apparent emergence of T. gondii in southern sea otters, we compiled environmental, demographic and serological data from 223 live and dead sea otters examined between 1997 and 2001. The T. gondii seroprevalence was 42% (49/116) for live otters, and 62% (66/107) for dead otters. Demographic and environmental data were examined for associations with T. gondii seropositivity, with the ultimate goal of identifying spatial clusters and demographic and environmental risk factors for T. gondii infection. Spatial analysis revealed clusters of T. gondii-seropositive sea otters at two locations along the coast, and one site with lower than expected T. gondii seroprevalence. Risk factors that were positively associated with T. gondii seropositivity in logistic regression analysis included male gender, older age and otters sampled from the Morro Bay region of California. Most importantly, otters sampled near areas of maximal freshwater runoff were approximately three times more likely to be seropositive to T. gondii than otters sampled in areas of low flow. No association was found between seropositivity to T. gondii and human population density or exposure to sewage. This study provides evidence implicating land-based surface runoff as a source of T. gondii infection for marine mammals, specifically sea otters, and provides a convincing illustration of pathogen pollution in the marine ecosystem. PMID- 12076630 TI - Cryptosporidium molnari n. sp. (Apicomplexa: Cryptosporidiidae) infecting two marine fish species, Sparus aurata L. and Dicentrarchus labrax L. AB - Cryptosporidium molnari n. sp. is described from two teleost fish, the gilthead sea bream (Sparus aurata L.) and the European sea bass (Dicentrarchus labrax L.). The parasite was found mainly in the stomach epithelium and seldom in the intestine. Oocysts were almost spherical, with four naked sporozoites and a prominent residuum, and measured 3.23-5.45 x 3.02-5.04 (mean 4.72 x 4.47) microm in the type host, gilthead sea bream (shape index 1-1.17, mean 1.05). Sporulation was endogenous, as fully sporulated oocysts were found within the fish, both in the stomach epithelium and lumen, and in faeces. Oocysts and other stages of C. molnari fit most of the diagnostic features of the genus Cryptosporidium, but differ from hitherto described species, including piscine ones. All stages were located within a host contributed parasitophorous vacuole lined by a double host microvillar membrane. Merogonial and gamogonial stages appeared in the typical extracytoplasmic position, whereas oogonial and sporogonial stages were located deeply within the epithelium. Ultrastructural features, including the characteristic contact zone of the parasite with the host epithelial surface, were mostly coincident with those of other Cryptosporidium spp. Mitochondria were found in dividing meronts, merozoites, microgamonts and sporozoites. Pathological effects were more evident in gilthead sea bream, which also exhibited a clearly higher prevalence (24.4 versus 4.64% in sea bass). External clinical signs, consisting of whitish faeces, abdominal swelling and ascites, were rarely observed, in contrast with important histopathological damage. The wide zones of epithelium invaded by oogonial and sporogonial stages appeared necrotic, with abundant cell debris, and sloughing of epithelial cells, which detached to the lumen. No inflammation reaction was observed and the cellular reaction was limited to the cells involved in the engulfing of intraepithelial stages and debris, probably macrophages. PMID- 12076631 TI - Sequence analysis of the beta-giardin gene and development of a polymerase chain reaction-restriction fragment length polymorphism assay to genotype Giardia duodenalis cysts from human faecal samples. AB - The flagellate parasite Giardia duodenalis is a major cause of diarrhoea in humans and in animals worldwide. Molecular techniques are particularly useful for studying the taxonomy, the population structure, the zoonotic potential of animal isolates, and the correlation between the genetic variability of the parasite and the range of clinical symptoms observed in humans. In this work, a new PCR assay that targets the beta-giardin gene was tested on 21 Giardia duodenalis reference strains representing Assemblages A, B and E, which are associated with infections of humans and other mammals. The assay was then applied to 30 faecal samples collected from Italian persons. The sequence analysis of 31 PCR products from both reference strains and clinical samples showed that each Assemblage is clearly distinct from the others on the basis of specific substitutions; the sequence diversity was approximately 5%, and all substitutions occurred at the third codon positions of the gene. The analysis of the intra-Assemblage variability allowed for the identification of three genotypes within Assemblage A, and of four genotypes within Assemblage B. Interestingly, two genotypes were identified only in the clinical samples and not in reference strains. Finally, a simple PCR-restriction fragment length polymorphism method was developed for the rapid discrimination of Assemblages and applied for the direct genetic analysis of cysts present in human faecal samples. PMID- 12076632 TI - Characterisation and expression of the Fasciola gigantica cathepsin L gene. AB - The gene structure of a cathepsin L from Fasciola gigantica was characterised. The gene spans approximately 2.0 kb and comprises four exons and three introns and is a compact gene as in the cases of crustaceous and platyhelminth cathepsins L. Southern blot analysis suggested that a few copies of the genes are sparsely organised in the genome. Of the three intron insertion positions, two of which are in the same position as in the mammalian cathepsin L gene. Phylogenetic analysis revealed that F. gigantica cathepsin L forms a clade with those from Fasciola hepatica, but not with those from Spirometra erinacei and schistosomes. Putative TATA-boxes were found upstream of a transcription initiation site. The sequence analysis of the 5'-upstream of the transcript revealed that the cathepsin L gene is transcribed by cis-splicing fashion. Furthermore, the experiments using recombinant F. gigantica procathepsin L showed that it was processed to an enzymatically active cathepsin L by pH-dependent autocatalysis. However, the pro-peptide deleted cathepsin L showed no enzyme activity, indicating that the pro-region of F. gigantica procathepsin L is essential for the folding and/or refolding of functional cathepsin L. These results are consistent with the observations in mammalian cathepsin L and papain. PMID- 12076633 TI - Characterisation of atovaquone resistance in Leishmania infantum promastigotes. AB - Atovaquone, an antiparasitic agent, could possibly represent an alternative therapy after relapse following classical treatment for visceral leishmaniasis. Atovaquone-resistant strains were selected in vitro by stepwise drug pressure to study the mechanism of resistance in Leishmania. Characteristics of a promastigote strain resistant to 250 microg/ml of atovaquone were compared with those of the wild type (WT) strain. Resistant strains were shown to have a high level of resistance (45 times). They were stable in drug-free medium for 6 months, and showed no cross-resistance with other antileishmanial drugs. Rhodamine uptake and efflux were studied. They were not modified in the resistant strain, indicating the absence of P-glycoprotein overexpession. The effect of atovaquone on membrane lipidic composition was determined in both WT and atovaquone-resistant promastigotes. Analysis of lipid composition of the atovaquone-resistant strain showed that sterol biosynthesis was decreased in atovaquone-resistant parasites. Cholesterol was found to be the major membrane sterol as opposed to the WT strain. Cholesterol, due to its ordering effect, could decrease membrane fluidity and subsequently block the passage of atovaquone through the membrane. Increased membrane cholesterol content and altered drug membrane fluidity resulted from possible decrease of ergosterol biosynthesis by atovaquone, incorporation of cholesterol by promastigotes in the culture medium, solubilisation of atovaquone by cholesterol and co-passage of the two compounds or influence of dimethylsulfoxide. These results indicate that different cellular alterations may participate in the resistant phenotype, by altering drug membrane permeability. PMID- 12076634 TI - Establishment of Besnoitia darlingi from opossums (Didelphis virginiana) in experimental intermediate and definitive hosts, propagation in cell culture, and description of ultrastructural and genetic characteristics. AB - Besnoitia darlingi from naturally infected opossums (Didelphis virginiana) from Mississippi, USA, was propagated experimentally in mice, cats, and cell culture and was characterised according to ultrastructural, genetic, and life-history characteristics. Cats fed tissue cysts from opossums shed oocysts with a prepatent period of nine or 11 days. Oocysts, bradyzoites, or tachyzoites were infective to outbred and interferon-gamma gene knockout mice. Tachyzoites were successfully cultivated and maintained in vitro in bovine monocytes and African green monkey cells and revived after an 18-month storage in liquid nitrogen. Schizonts were seen in the small intestinal lamina propria of cats fed experimentally-infected mouse tissues. These schizonts measured up to 45 x 25 microm and contained many merozoites. A few schizonts were present in mesenteric lymph nodes and livers of cats fed tissue cysts. Ultrastructurally, tachyzoites and bradyzoites of B. darlingi were similar to other species of Besnoitia. A close relationship to B. besnoiti and an even closer relationship to B. jellisoni was indicated for B. darlingi on the basis of the small subunit and ITS-1 portions of nuclear ribosomal DNA. PMID- 12076635 TI - How to improve statistical analysis in parasitology research publications. PMID- 12076636 TI - Toxic effects of head-to-tail 3-alkylpyridinium polymers isolated from the marine sponge Reniera sarai in rat. AB - Toxic water soluble polymeric 3-alkylpyridinium salts (poly APS; MW 18900 and 5520Da) were isolated from the marine sponge Raniera sarai. In vitro they strongly inhibited acetylcholinesterase. In order to evaluate the role of acetylcholinesterase inhibition in toxin lethality, and to assess other possible lethal effects, in vivo experiments were performed on male Wistar rats, and ECG, blood pressure and breathing pattern were monitored. The results showed that none of the animals died due to the acetylcholinesterase inhibitory action of poly APS. Doses lower than 1mg/kg caused only transient bradycardia and transient prolongation of expirium. At doses above 2.7mg/kg of poly-APS all treated animals died, but signs were not typical of acetylcholinesterase inhibition. Arterial blood pressure fell to mid-circulatory pressure, and breathing stopped after a few breaths with an increase of the residual volume. Autopsy of the experimental animals that died due to the effects of the toxin revealed that mid-size and small sized blood vessels in the heart and lungs were filled with granular brownish material with inclusions of red blood cells and platelets. Data obtained on blood samples from animals treated with poly-APS also revealed numerous thrombocyte aggregates. In vitro poly-APS induced thrombocyte aggregation in a dose dependent manner. The acetylcholinesterase-inhibitory effects were most pronounced only at lower doses of poly-APS. With higher doses those effects were masked or covered by other, more pronounced and faster developing lethal effects of the toxin such as platelet aggregation. Therefore it is reasonable to assume that acetylcholinesterase inhibitory effects are not responsible for the lethal activity of the toxin. PMID- 12076637 TI - Gossypol, a component in cottonseed, induced increases in cytosolic Ca2+ levels in Chang liver cells. AB - The effect of gossypol, a compound found in cottonseed, on intracellular free Ca2+ levels ([Ca2+](i)) in Chang liver cells were evaluated using fura-2 as a fluorescent Ca2+ indicator. Gossypol (0.2-5microM) increased [Ca2+](i) in a concentration-dependent manner with an EC(50) value of 1.5microM. The [Ca2+](i) response was composed of an initial rise and a slow decay to a sustained phase within 5min after drug application. Removal of extracellular Ca2+ markedly reduced the [Ca2+](i) signals by 80+/-2%. Preincubation with 0.1mM La3+ or 10microM nimodipine abolished the Ca2+ influx. Gossypol (5microM)-induced release of intracellular Ca2+ was reduced by 75% by pretreatment with 1microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor) to deplete the endoplasmic reticulum Ca2+. Conversely, pretreatment with gossypol abolished thapsigargin-induced Ca2+ release. After pretreatment with 5microM gossypol in Ca2+-free medium for several min, addition of 3mM Ca2+ induced a [Ca2+](i) increase of a magnitude nine-fold greater than control. Gossypol (5microM) induced Ca2+ release was not affected by inhibiting phospholipase C with 2microM 1-(6-((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5 dione (U73122). Together, this study shows that gossypol induced significant [Ca2+](i) increases in Chang liver cells by releasing Ca2+ from intracellular pools in a phospholipase C-dissociated fashion and by causing La3+- and nimodipine-sensitive Ca2+ influx. PMID- 12076638 TI - Development of viper-venom antibodies in chicken egg yolk and assay of their antigen binding capacity. AB - The therapeutic use of specific antibodies is invaluable in certain clinical conditions, such as administration of specific antivenom for snakebite envenomation. The production of antibodies and their purification from mammalian blood has been found low yielding and laborious. Most antivenom is polyvalent whole serum or partially purified immunoglobulin. The side effects of anti-snake venom therapy include serum sickness and can be reduced by using mono-specific antivenom in sufficiently pure form. We have attempted to standardize a simple method for producing avian antivenom in relatively pure form from eggs. The isolation is very simple and involves only two steps, namely, removal of lipids from the diluted egg yolk followed by gel filtration. Each egg produces 80-100mg of pure immunoglobulin, and specific antibodies are present for up to 100 days after immunization. Thus, large quantities of the Ig can be obtained in pure form using only small amounts of venom. Antigen binding was shown by Ouchterlony's double diffusion experiments and the avian antivenom neutralizes the thrombin like activity of equivalent amounts of venom on human plasma. The LD(50) of the venom was approximately 3mg/kg body weight in mice and rats but when pre incubated with equivalent amounts (by weight) of egg IgG injected subcutaneously, all the animals survived. In a similar experiment using a commercial horse IgG, 25% mortality is seen. These results indicate that the antivenom immunoglobulins purified from immunized chicken egg yolk is biologically active and the possibility of their therapeutic use will be investigated further. PMID- 12076639 TI - Pharmacological characterization of mikatoxin, an alpha-neurotoxin isolated from the venom of the New-Guinean small-eyed snake Micropechis ikaheka. AB - Symptoms of envenomation by the New-Guinean small-eyed snake Micropechis ikaheka (Elapidae) include peripheral neurotoxicity and myotoxicity. We have now purified to homogeneity a long-chain neurotoxin, mikatoxin, from M. ikaheka venom by successive gel filtration and reverse-phase chromatography. Electrospray ionization mass spectrometry showed mikatoxin to be a homogenous peptide of MW 7775.6. Mikatoxin was devoid of any phospholipase A(2) activity associated with the crude venom and did not exhibit any intrinsic anticholinesterase activity. In the chick biventer cervicis muscle, it produced an irreversible, concentration dependent block of responses to exogenously applied acetylcholine and carbachol as well as twitches evoked by nerve, but not by direct muscle stimulation. Moreover, mikatoxin, like alpha-bungarotoxin and erabutoxin-b, did not show significant fade response to train-of-four stimulation of the mouse phrenic nerve hemi diaphragm muscle. It also failed to block ganglionic transmission in the guinea pig ileum and muscarinic responses in the rat anococcygeus muscle. Our study provides strong evidence for the presence of a neurotoxin (mikatoxin) in M. ikaheka venom that produces neuromuscular blockade in skeletal muscle attributable to selective and irreversible antagonism of postsynaptic nicotinic acetylcholine receptors of the neuromuscular junction and likely contributes to the peripheral neurotoxicity observed in M. ikaheka envenomation. PMID- 12076640 TI - A comparative analysis of invaded sequences from group IA phospholipase A(2) genes provides evidence about the divergence period of genes groups and snake families. AB - Two phospholipase A(2) (PLA(2)) genes classified into group IA were cloned from the genomic library of the sea snake Laticauda semifasciata. Eight clones were obtained by PCR cloning procedure from genomic DNA of Laticauda laticaudata (four clones) and Laticauda colubrina (four clones). The genes were 3.6-4.4kbp in length. Intron and exon organization of the group IA PLA(2) genes was the same as that of Naja sputatrix group IA PLA(2) genes (four exons and three introns). There were two kinds of repetitive sequences in the first and second introns of all sequenced PLA(2) genes. The differences in the length of these genes were derived from the length of their repetitive sequences. The chicken repeat-1 (CR1) like long interspersed repeated DNA (LINE) sequences, different from the above repetitive sequences, were also found in all sequenced Laticauda PLA(2) genes. A comparative analysis of groups IA, IA' and IIA PLA(2)s genes suggests a period of CR1-like LINE integration during molecular and family evolution. The integration of CR1-like LINE into PLA(2) genes occurred after the divergence of groups I and II PLA(2)s but before the divergence of groups, IA and IA' PLA(2)s. These integration events occurred before the family divergence of Naja and Laticauda. The presence of CR1-like LINE and a comparison of intron and exon organization showed that the divergence of Naja and Bungarus occurred before the divergence of Laticauda and Naja. PMID- 12076641 TI - Antibacterial activity against resistant bacteria and cytotoxicity of four alkaloid toxins isolated from the marine sponge Arenosclera brasiliensis. AB - Arenosclerins A-C and haliclonacyclamine E, new tetracyclic alkylpiperidine alkaloids isolated from the marine sponge Arenosclera brasiliensis, were subjected to antimicrobial and cytotoxic bioassays. Fourteen samples of microorganisms were used: Candida albicans, Staphylococcus aureus, Escherichia coli, and 12 antibiotic-resistant bacteria isolated from hospital environment. The minimum inhibitory concentration activity of each alkaloid was determined. The four compounds displayed antibacterial activity, but no antifungal activity against C. albicans. Haliclonacyclamine E and arenosclerins A and C were active against a larger number of bacteria strains than arenosclerin B. However, arenosclerins B and C presented more potent antibacterial activity. The alkaloids displayed inhibitory activity against both Gram positive and Gram negative bacteria. Cytotoxicity bioassays using the MTT method showed that these compounds present cytotoxic activity against human HL-60 (leukemia), L929 (fibrosarcoma), B16 (melanoma) and U138 (colon) cancer cell lines at concentrations between 1.5 and 7.0microg/ml. The results obtained indicated that A. brasiliensis alkaloids have a potent toxic activity. The broad cytotoxic and antimicrobial activities presented by A. brasiliensis alkaloids suggest a defensive role of arenosclerins and haliclonacyclamine E against microbial infection and/or the action of potential predators at the sponge's natural habitat. PMID- 12076642 TI - Determination of microcystins in lake water using reusable immunoaffinity column. AB - A reusable immunoaffinity column for purification of microcystins in lake water was prepared by coupling anti-microcystin-LR monoclonal antibodies to immunoaffinity support. Thanks to spherical shape of the immunoaffinity support Formyl-Cellulofine used in this study, applied solutions passed the column smoothly even when used repeatedly. Reusability of the column was examined by determining the recoveries of spiked microcystins-RR, -YR and -LR (100ng each) from lake water. After extraction with a Sep-Pak PS2 cartridge containing styrene divinylbenzene copolymer, the extract was purified with the immunoaffinity column. The immunoaffinity column was regenerated by washing with Tris-HCl buffer containing bovine serum albumin for repeated uses. Recoveries of spiked microcystins from the first use of the column were 87-88%, and 83-88% from the second and third uses, and the recoveries gradually dropped to 63-77% from the 4 5th uses, the results of which indicated that the column could be used repeatedly for three times. The present method was applied to determine microcystins in water collected from three different lakes in Japan in 1999. In a sample from Lake Suwa, microcystins-RR and -LR were determined by high performance liquid chromatography with photodiode array detection and electrospray ionization-liquid chromatography/mass spectrometry. PMID- 12076643 TI - Conophysin-R, a Conus radiatus venom peptide belonging to the neurophysin family. AB - A novel Conus peptide, conophysin-R, was purified from the venom of Conus radiatus. The distinctive disulfide framework and sequence indicates that it is a member of the neurophysin peptide family. The complete sequence of the peptide is HPTKPCMYCSFGQCVGPHICCGPTGCEMGTAEANMCSEEDEDPIPCQVFGSDCALNNPDNIHGHCVADGICCVDDTCTTHL CLThis is the first time a neurophysin-like peptide has been found in any venom. In addition, conophysin-R is the first neurophysin family member isolated and biochemically characterized from an invertebrate source. PMID- 12076644 TI - First evidence of an extensive northern European distribution of azaspiracid poisoning (AZP) toxins in shellfish. AB - Azaspiracids have recently been identified as the toxins responsible for a series of human intoxications in Europe since 1995, following the consumption of cultured mussels (Mytilus edulis) from the west coast of Ireland. Liquid chromatography-mass spectrometric (LC-MS) methods have been applied in the study reported here to investigate the new human toxic syndrome, azaspiracid poisoning. Separation of azaspiracid (AZA1) and its analogues, 8-methylazaspiracid (AZA2) and 22-demethylazaspiracid (AZA3), was achieved using reversed-phase LC and coupled, via an electrospray ionisation source, to an ion-trap mass spectrometer. These azaspiracids have now been identified in mussels from Craster (north-east England) and Sognefjord (south-west Norway) using source collision induced dissociation-MS and multiple tandem MS detection. AZA1 was the predominant toxin and toxin profiles were similar to those found in contaminated Irish shellfish. This is the first report of the occurrence of these azaspiracids outside Ireland with the significant implications that these toxins may occur in shellfish throughout northern Europe. PMID- 12076645 TI - Crystal structures of an acidic phospholipase A(2) from the venom of Naja Kaouthia. AB - A phospholipase A(2) purified from the venom of Naja kaouthia (Guangxi cobra) exhibits anticoagulant activities. The structures of two crystal forms were determined by X-ray crystallography at 2.8A resolution with the Naja naja (India cobra) PLA(2) as an initial model. The enzyme exhibits a trimer structure, which is similar to that of India cobra PLA(2). This reinforces the physiological relevance of the oligomer. The trimer has a wide cavity, which allows the substrate to enter and interact with the catalytic site. The formation of the trimer may serve as a storage method to improve the solubility at high concentration in the venom gland. The Ca2+ binding loop in the absence of the cation can exist in different conformations depending on its surroundings. PMID- 12076646 TI - Botulinum neurotoxin serotypes A and B preparations have different safety margins in preclinical models of muscle weakening efficacy and systemic safety. AB - This preclinical study compared the muscle weakening efficacy, duration, and safety margin of the recently approved botulinum toxin type B (BTX-B; Myobloc/Neurobloc) to botulinum toxin type A (BTX-A; BOTOX((R))). Mice received a single hind limb intramuscular injection of BTX-B (1-150U/kg) or BTX-A (1 120U/kg). An observer who was masked to treatment assessed the magnitude and duration of muscle weakening efficacy on a 0-4 scale using the digit abduction scoring assay. Safety margins were determined as the ratio of the IM median lethal dose to the IM dose that produced half-maximal muscle weakness in the DAS. BTX-A produced muscle weakness at lower doses than BTX-B (IM ED(50): 6.2+/-0.6 vs. 20.8+/-1.4U/kg, respectively) (p<0.0001). BTX-A at 29U/kg and BTX-B at 67U/kg produced comparable peak DAS scores of approximately 4 indicating maximal muscle weakness. At these doses, the duration of BTX-A was longer, with a return to baseline by day 36 compared to a return to baseline by day 14 with BTX-B. The mean dose that was lethal in 50% of mice was lower for BTX-A than BTX-B (81.4+/ 3.5 vs. 104.6+/-1.9U/kg, respectively) (p<0.001) and the safety margin was higher (13.9+/-1.7 vs. 5.4+/-0.3, respectively (p<0.001). These results indicate that the BTX-A:BTX-B dose ratio for muscle weakening efficacy is different from the ratio for systemic effects following IM injections and suggest that no single dose ratio is adequate to compare these preparations. The in vivo differences found are consistent with the different clinical profiles reported for these two products. PMID- 12076647 TI - Characterisation of multiple Caribbean ciguatoxins and congeners in individual specimens of horse-eye jack (Caranx latus) by high-performance liquid chromatography/mass spectrometry. AB - We studied the variation in toxin profiles of purified extracts of 10 individual specimens and two pools of ciguateric Caranx latus. High-performance liquid chromatography/mass spectrometry (HPLC/MS) identified in all individual samples at least seven Caribbean ciguatoxins (C-CTXs) comprising C-CTX-1 and its epimer C CTX-2 ([M+H](+) m/z 1141.58), and five new C-CTX congeners with pseudo-molecular ions at m/z 1141.58, 1143.60, 1157.57, 1159.58, and 1127.57. In some samples, additional C-CTX isomers were detected with [M+H](+) ions at m/z 1141.58 (two), 1143.60 (one) and 1157.57 (two). The two low-toxic pools contained only four to six ciguatoxins. The comparison in relative proportions of four different mass classes ([M+H](+) at m/z 1141, 1143, 1157 and 1127) showed that the group at m/z 1157 increased (2-20%) with flesh toxicity. More than 80% of group m/z 1141 comprised C-CTX-1, C-CTX-2 and their isomer C-CTX-1a whose level in this group correlated with fish toxicity. Contrary to low-toxic fishes, high-risk specimens had C-CTX-1 levels <50% and were subjected to large losses of activity on purification indicating that unstable ciguatoxins were present. A possible conversion of C-CTX-1 into C-CTX-1a was identified when flesh was cooked, without changes in toxicity. In conclusion, HPLC/MS characterised 12 C-CTXs accumulated by C. latus at variable levels. PMID- 12076648 TI - Glutathione-S-transferase activity toward aflatoxin epoxide in livers of mastomys and other rodents. AB - In order to study the liver glutathione-S-transferase (GST) activity toward aflatoxin B1 (AFB1) epoxide in mastomys in comparison with other rodents, we performed in vitro studies of the cytosolic GST activity toward AFB1-epoxide using mastomys, rat, mouse and hamster liver. Also AFB1 metabolism by liver microsomes including formation of AFB1-DNA adducts was studied. Cytosolic GST activity toward AFB1-epoxide was highest in mastomys liver, and higher in the hamster and mouse livers than in the rat liver, correlating well with the differences of the sensitivity of these species to the toxicity of AFB1. However, no relationship was noted between the sensitivity of a given species to the toxicity of AFB1 and the microsomal activity of binding of AFB1 to DNA or metabolizing AFB1 to AFM1, AFQ1 and AFP1. These results demonstrate the importance of the GST mediated AFB1-epoxide conjugation with glutathione in determining the differing sensitivities of these species to AFB1 toxicity. The extremely high activity of GST in mastomys indicates that this species would be a good model animal for studying GST toward AFB1-epoxide. PMID- 12076649 TI - Characterization and cDNA cloning of halyxin, a heterogeneous three-chain anticoagulant protein from the venom of Agkistrodon halys brevicaudus. AB - We report upon the isolation, characterization, and cDNA cloning of an anticoagulant protein, halyxin from Agkistrodon halys brevicaudus venom. The protein exists as a 29kDa protein, and is separated into three chains on SDS-PAGE under reducing conditions. However, we cloned only two cDNAs encoding halyxin from the cDNA library of the snake venom gland, on the basis of the determined amino acid sequences. The complete amino acid sequences were deduced from their nucleotide sequences and named halyxin A (129 amino acid residues) and B chain (123 amino acid residues). The deduced amino acid sequence of halyxin A chain corresponds to the two smaller chains. Thus, it is considered that halyxin A chain could be synthesized as a single-chain protein that is subsequently cleaved to yield the mature two-chain protein. The amino acid sequence of halyxin is similar to that of other snake venom proteins of the C-type lectin superfamily, and prolongs plasma-clotting time. In the presence of Ca(2+) ions, halyxin binds to coagulation factors IX, X, IXa, and Xa, but not to other vitamin K-dependent coagulation factors. It also inhibits factor Xa in a non-competitive manner but does not affect other activated coagulation factors. PMID- 12076650 TI - Amino acid sequence of a thrombin like enzyme, elegaxobin, from the venom of Trimeresurus elegans (Sakishima-habu). AB - The amino acid sequence of a thrombin like enzyme, named elegaxobin, isolated from the venom of Trimeresurus elegans (Sakishima-habu) was determined by Edman sequencing of the peptides, derived from digests with cyanogen bromide, hydroxylamine, achromobacter protease I, trypsin, V8 protease, and chymotrypsin. Elegaxobin showed conservation of the catalytic amino acid residues (His, Asp, and Ser) of trypsin like serine protease in the amino acid sequence. The carboxy terminal amino acid, Leu, was determined using carboxypeptidase Y. Elegaxobin consisted of 233 amino acids and had a calculated molecular weight of 25,439. Elegaxobin was 53, 59, 26 and 40% homologous in sequence to ancrod, flavoxobin, bovine thrombin and trypsin, respectively. PMID- 12076651 TI - From sanddabs to blue whales: the pervasiveness of domoic acid. AB - Domoic acid (DA) is a potent food web transferred algal toxin that has caused dramatic mortality events involving sea birds and sea lions. Although no confirmed DA toxicity events have been reported in whales, here we present data demonstrating that humpback and blue whales are exposed to the toxin and consume DA contaminated prey. Whale fecal samples were found to contain DA at levels ranging from 10 to 207microg DA g(-1) feces via HPLC-UV methods. SEM analysis of whale feces containing DA, collected from krill-feeding whales, revealed the presence of diatom frustules identified as Pseudo-nitzschia australis, a known DA producer. Humpback whales were observed feeding on anchovies and sardines that contained DA at levels ranging from 75 to 444microg DA g(-1) viscera. DA contamination of whale feces and fish occurred only during blooms of toxic Pseudo nitzschia. Additionally, several novel fish species collected during a toxic diatom bloom were tested for DA. Fish as diverse as benthic sanddabs and pelagic albacore were found to contain the neurotoxin, suggesting that DA permeates benthic as well as pelagic communities. PMID- 12076652 TI - Pectenotoxin-2 seco acid, 7-epi-pectenotoxin-2 seco acid and pectenotoxin-2 in shellfish and plankton from Portugal. AB - Pectenotoxin-2 seco acid (PTX2sa) and 7-epi-pectenotoxin-2 seco acid (7-epi PTX2sa) were found in Portuguese shellfish both by fluorescence detection after ADAM derivatisation and, liquid chromatography coupled with mass spectrometry detection. Two time-series both with blue mussel (Mytilus edulis) and common cockle (Cerastoderma edule) from Aveiro lagoon illustrate how PTX2sa has a strong association with Dinophysis acuta occurrence in the plankton, as well as Dinophysis fortii. Data so far excludes D. acuminata from contributing to contamination with pectenotoxins. It also shows that mussel may not be the best indicator of contamination with PTX2sa. At Aveiro lagoon also oyster, razor clam and clams were all less toxic than cockle. Pectenotoxin-2 seco acid was not involved in a previously reported incident of human poisoning that took place in February 1998 after consumption of Donax trunculus. In plankton extracts the most abundant pectenotoxin found was PTX2. Concentration of PTX2sa was around 10% of PTX2, and 7-epi-PTX2sa was not detected. PMID- 12076653 TI - First confirmation of human diarrhoeic poisonings by okadaic acid esters after ingestion of razor clams (Solen marginatus) and green crabs (Carcinus maenas) in Aveiro lagoon, Portugal and detection of okadaic acid esters in phytoplankton. AB - A new outbreak of human diarrhoeic poisonings (DSP) with esters of okadaic acid (OA) was confirmed after ingestion of razor clams (Solen marginatus) harvested at Aveiro lagoon (NW Portugal) in the summer of 2001. Accumulation of marine toxins in second order consumers was investigated in the edible parts of a shellfish predator abundant at Aveiro lagoon, the green crab Carcinus maenas. Okadaic acid was found, also in a predominant esterified form. Levels in edible parts (comprising mainly viscera) surpassed 16microg/100g. We suggest that one patient may have developed profuse diarrhoea after ingestion of a large number of green crabs contaminated with okadaic acid esters. At least 32microg OA/100g were found in a remaining sample of its meal. Domoic acid was also found but under the allowable level in force in USA of 30microg/g crab viscera. In cooked crabs, significant losses of domoic acid were found and it is not suspected to have contributed to the poisoning event, although being a vector for this toxin. The low percentage of free okadaic acid found is in accordance with a predation predominantly on benthonic shellfish (razor clams, clams and common cockle) rather than on rock mussels. These last ones present usually higher percentages of free okadaic acid. Okadaic acid was confirmed with full-scan mass spectra either in plankton and mussel extracts. Okadaic acid esters were also found in plankton extracts. Percentages between 40-60% of esterified OA were found in samples freshly extracted. Ester's percentage diminished drastically if after sonication the extract was kept at room temperature. The major part of the esters was water-soluble. PMID- 12076654 TI - Ontogenetic variation of metalloproteinases and plasma coagulant activity in venoms of wild Bothrops atrox specimens from Amazonian rain forest. AB - A comparative study of venoms from juvenile, sub-adult and adult wild Bothrops atrox specimens captured in Manaus region (Brazil) was performed. All venoms tested had acidic pH (5.5) and the human plasma coagulant activity was higher in venoms from juvenile and sub-adult specimens than in adults. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the most intense bands in adult venoms corresponded to polypeptides of 23 and 50kDa. The 23kDa protein was not detected in juvenile venoms. The 23 and 50kDa proteins were purified by two steps of reversed phase-HPLC followed by size exclusion HPLC. Partial amino acid sequence of the 23kDa protein showed homology to metalloproteinases from other snake venoms. Electrospray ionization mass spectrometric analysis (ESI-MS) showed that the 23kDa band contained at least three isoforms of 23030, 23300 and 23645Da. The 50kDa polypeptide was N terminally blocked for Edman degradation and presented molecular masses ranging from 46.8 to 49.4kDa by ESI-MS. Both proteins were detected by anti-mutalysin II antibodies in immunoblotting assay indicating that they belong to the metalloproteinase family. Immunoblotting analysis also showed that the 23kDa band increased in intensity from juvenile to adult specimens.SDS-PAGE analysis of juvenile and adult venoms following autoproteolysis in pH 7.4 suggested that endogenous venom metalloproteinases can digest the 50kDa metalloproteinase, originating a new protein band of 27kDa. It was also demonstrated in juvenile venoms that the 23kDa band was not the result of proteolytic processing of the 50kDa metalloproteinase. PMID- 12076655 TI - Pathogenic mechanisms of sand tampan toxicoses induced by the tick, Ornithodoros savignyi. AB - The tick, Ornithodoros savignyi has been implicated in inducing paralysis and tampan toxicosis. In this study, a basic toxin (TSGP4) was identified and the presence of an acidic toxin (TSGP2) was confirmed. Both basic and acidic toxins were more lethal than previously described, with TSGP4 (34microg) and TSGP2 (24microg) causing mortality of adult mice within 30min. Pathological effects on the cardiac system, notably of salivary gland extract on an isolated rat heart perfusion system and of purified toxins on mouse electrocardiogram patterns could be observed. TSGP4 caused Mobitz type ventricular block, while TSGP2 induced ventricular tachycardia. Conversely, fractions from reversed phase high performance liquid chromatography preparations caused paralysis-like symptoms of the limbs after only 48h. The toxins also differ from previously described tick paralysis toxins in terms of molecular behavior and properties. These results indicate that tampan toxicoses and tick paralysis are unrelated pathogenic phenomena. PMID- 12076657 TI - Study of experimentally induced lesions in sheep by grazing Brachiaria decumbens. AB - A histologic and ultrastructural study of the alterations found in the lymph nodes and livers of nine sheep with experimental cholangiohepatopathy by grazing on Brachiaria decumbens has been performed. Sheep were euthanized in three groups, on the 77th, 89th, and 150th days of the experimental feeding. The main gross lesions were whitish spots of multifocal distribution scattered throughout the hepatic parenchyma from all B. decumbens-grazed animals and whitish foci surrounded by reddened halos in the mesenteric and hepatic lymph nodes of sheep necropsied on the 150th. The principal histologic findings included hepatocellular cloudy swelling, marked multifocal cholangitis in the portal triads with bile duct proliferation and infiltration of macrophages and lymphocytes. Crystals were observed within bile ducts and surrounded by macrophages. Ultrastructurally, there were criytaloid structures within the macrophages and hepatocytes, which also presented hyperplasia of smooth endoplasmic reticulum. These findings suggest that hepatocytes were the initial target of the toxic effects, which depending on the degree of severity developed would cause both, subsequent cholangiopathy or occasional photosensitization. Additionally, the developmental stages of the hepatic lesions observed in this study have been presented. PMID- 12076656 TI - Comparison of protein phosphatase inhibitory activity and apparent toxicity of microcystins and related compounds. AB - Two metabolites of microcystin-LR glutathione conjugate and, microcystin-cysteine conjugate, as well as microcystin-RR (MCRR) are less toxic than microcystin-LR (MCLR). In the present study, we investigated why these compounds are weakly toxic in comparison with MCLR, as the reason is still unknown and no systematic study has so far been carried out for a clarification of this issue. Although they showed almost the same inhibitory activity against protein phosphatases 1 and 2A as MCLR in vitro, the apparent toxicity of these three compounds by intratracheal administration to mice decreased to about 1/12 the level of MCLR at 100microg/kg. An immunostaining study showed that these conjugates at a sublethal dose of 200microg/kg were prominently observed in the intestine and kidney, whereas effective accumulation and bleeding were not found in the liver in spite of the larger dosage. As an explanation for these results, there may be two possibilities. First, the transport system to the liver might not function well, and second, transported toxins may be effectively eliminated by an appropriate system such as the GS-X (ATP-dependent glutathione S-conjugate exported) pump. It was concluded that the inhibitory activity against protein phosphatases is not always related to the apparent LD(50) level, and that the appearance of toxicity by microcystins depends on the balance between accumulation and metabolism in the liver. PMID- 12076659 TI - Enzymes with gelatinolytic activity can be found in Tityus bahiensis and Tityus serrulatus venoms. AB - Enzymes with gelatinolytic activity were detected in Tityus bahiensis and Tityus serrulatus venom. Their activity was optimal at pH 8.0 in SDS-PAGE-gelatin. They were inhibited by PMSF but not by iodoacetamide, pepstatin or phenantrolin in the assay conditions used. This suggests that these enzymes are serine proteases. The presence of metal ions did not affect the proteolytic activity of these enzymes. Several possible functions may be envisaged for these enzymes: in tissue permeabilization, pancreatitis and toxin processing. PMID- 12076658 TI - Molecular cloning and sequence analysis of cDNA encoding flavoridin, a disintegrin from the venom of Trimeresurus flavoviridis. AB - We isolated a cDNA of 2001bp encoding the full-length precursor of flavoridin, which is one of the four disintegrins in the venom of Trimeresurus flavoviridis, and analyzed the cDNA nucleotide sequence. The deduced amino acid sequence of the open reading frame consisted of a pro-domain (190 residues), a metalloproteinase domain (205 residues), a spacer domain (18 residues) and a disintegrin (flavoridin) domain (70 residues), thus indicating that the flavoridin precursor belongs to the P-II class of snake venom metalloproteinases. The unknown metalloproteinase domain shared strong sequence similarity with HR2a (71.2% identity) and H(2)-proteinase (74.1% identity), a low molecular mass hemorrhagic metalloproteinase and a non-hemorrhagic metalloproteinase in the same snake venom, respectively. PMID- 12076660 TI - Bradykinin is involved in hyperalgesia induced by Bothrops jararaca venom. AB - Bradykinin is involved in hyperalgesia (pain hypersensitivity) induced by Bothrops jararaca venom-intraplantar injection of B. jararaca venom (5microg/paw) in rats caused hyperalgesia, which peaked 1h after venom injection. This phenomenon was not modified by promethazine (H(1) receptor antagonist), methysergide (5-HT receptor antagonist), guanethidine (sympathetic function inhibitor), anti-TNF-alpha or anti-interleukin-1 antibodies or by the chelating agent CaNa(2)EDTA. Venom-induced hyperalgesia was blocked by the bradykinin B(2) receptor antagonist HOE 140. On the other hand, des-Arg(9), [Leu(8)]-bradykinin, a bradykinin B(1) receptor antagonist, did not modify the hyperalgesic response. These results suggest that bradykinin, acting on B(2) receptor, is a mediator of hyperalgesia induced by B. jararaca venom. PMID- 12076661 TI - A short-chain peptide toxin isolated from Centruroides sculpturatus scorpion venom inhibits ether-a-go-go-related gene K(+) channels. AB - From the venom of the American scorpion Centruroides sculpturatus Ewing we have isolated a minute peptide fraction (named CsEKerg1) which reversibly inhibits the current through ERG (ether-a-go-go-related gene) K(+) channels. Isolation was done by CM-cellulose column chromatography and reversed phase high-performance liquid chromatography. To test for an effect on ERG channels we used NG108-15 neuroblastomaxglioma hybrid cells voltage-clamped in the whole-cell mode. CsEKerg1 contains 43 amino acids and has a molecular weight of 4833. Its amino acid sequence is similar but not identical to that of ergtoxin, a peptide isolated recently from the venom of the Mexican scorpion Centruroides noxius [FASEB J. 13 (1999) 953]. Half inhibition of ERG current occurs at a peptide concentration of 1.12microg/ml. PMID- 12076663 TI - What is the rabbit skin method by Kondo et al. (1960) for determining hemorrhagic activities of snake venoms? PMID- 12076664 TI - Mushroom-induced rhabdomyolisis. Cortinarius or Tricholoma? PMID- 12076668 TI - Dominantly inherited, non-coding microsatellite expansion disorders. AB - Dominantly inherited diseases are generally caused by mutations resulting in gain of function protein alterations. However, a CTG expansion located in the 3' untranslated portion of a kinase gene was found to cause myotonic dystrophy type 1, a multisystemic dominantly inherited disorder. The recent discovery that an untranslated CCTG expansion causes the same constellation of clinical features in myotonic dystrophy type 2 (DM2), along with other recent discoveries on DM1 pathogenesis, have led to the understanding that both DM1 and DM2 mutations are pathogenic at the RNA level. These findings indicate the existence of a new category of disease wherein repeat expansions in RNA alter cellular function. Pathogenic repeat expansions in RNA may also be involved in spinocerebellar ataxia types 8, 10 and 12, and Huntington's disease-like type 2. PMID- 12076669 TI - Friedreich ataxia: a paradigm for mitochondrial diseases. AB - Friedreich ataxia (FRDA), a progressive neurodegenerative disease, is due to the partial loss of function of frataxin, a mitochondrial protein of unknown function. Loss of frataxin causes mitochondrial iron accumulation, deficiency in the activities of iron-sulfur (Fe-S) proteins, and increased oxidative stress. Mouse models for FRDA demonstrate that the Fe-S deficit precedes iron accumulation, suggesting that iron accumulation is a secondary event. Furthermore, increased oxidative stress in FRDA patients has been demonstrated, and in vitro experiments imply that the frataxin defect impairs early antioxidant defenses. These results taken together suggest that frataxin may function either in mitochondrial iron homeostasis, in Fe-S cluster biogenesis, or directly in the response to oxidative stress. It is clear, however, that the pathogenic mechanism in FRDA involves free-radical production and oxidative stress, a process that appears to be sensitive to antioxidant therapies. PMID- 12076670 TI - The fragile X premutation: into the phenotypic fold. AB - Premutation alleles (55-200 CGG repeats) of the fragile X mental retardation 1 gene (FMR1) are known to contribute to the fragile X phenotype through genetic instability and transmission of full mutation alleles (>200 repeats). There is now mounting evidence that the premutation alleles themselves contribute to clinical involvement, including premature ovarian failure among female carriers and a new tremor/ataxia syndrome among older male carriers. Recent observations also provide direct evidence of dysregulation of the FMR1 gene in the premutation range, which may explain many of the clinical observations. PMID- 12076671 TI - Advances in understanding of fragile X pathogenesis and FMRP function, and in identification of X linked mental retardation genes. AB - The fragile X mental retardation syndrome is caused by large methylated expansions of a CGG repeat in the FMR1 gene that lead to the loss of expression of FMRP, an RNA-binding protein. FMRP is proposed to act as a regulator of mRNA transport or translation that plays a role in synaptic maturation and function. The recent observations of unexpected phenotypes in some carriers of fragile X premutations suggest a pathological role, in these individuals, of an abnormal FMR1 mRNA. FMRP was recently shown to interact preferentially with mRNAs containing a G quartet structure. Mouse and Drosophila models are used to decipher the function of FMRP, which was found to inhibit translation of some mRNA targets, but may be stimulatory in other cases. Proteins interacting with FMRP have been identified, and suggest a link with the Rac1 GTPase pathway that is important in neuronal maturation. Recent advances also include identification of other genes implicated in X-linked mental retardation. PMID- 12076672 TI - The molecular bases of spinal muscular atrophy. AB - Spinal muscular atrophy (SMA) is a common recessive autosomal disorder characterized by degeneration of motor neurons of the spinal cord. SMA is caused by mutations of the survival of motor neuron gene that encodes a multifunctional protein, and mouse models have been generated. These advances represent starting points towards an understanding of the pathophysiology of this disease and the design of therapeutic strategies in SMA. PMID- 12076673 TI - Genetics of Parkinson's disease and biochemical studies of implicated gene products. AB - Parkinson's disease was thought, until recently, to have little or no genetic component. This notion has changed with the identification of three genes, and the mapping of five others, that are linked to rare familial forms of the disease (FPD). The products of the identified genes, alpha-synuclein (PARK 1), parkin (PARK 2), and ubiquitin-C-hydrolase-L1 (PARK 5) are the subject of intense cell biological and biochemical studies designed to elucidate the underlying mechanism of FPD pathogenesis. In addition, the complex genetics of idiopathic PD is beginning to be unraveled. Genetic information may prove to be useful in identifying new therapeutic targets and identifying the preclinical phase of PD, allowing treatment to begin sooner. PMID- 12076674 TI - Familial dysautonomia. AB - Familial dysautonomia is a developmental disorder of the sensory and autonomic nervous system. Recent studies have shown that two mutations in the gene IKBKAP are responsible for the disease. IKAP, the IKBKAP-encoded protein, is a member of the recently identified human Elongator complex. The major FD mutation is a splice mutation that results in aberrant tissue-specific mRNA splicing. PMID- 12076675 TI - Molecular-evolutionary mechanisms for genomic disorders. AB - Molecular studies of unstable regions in the human genome have identified region specific low-copy repeats (LCRs). Unlike highly repetitive sequences (e.g. Alus and LINEs), LCRs are usually of 10-400 kb in size and exhibit > or = 95-97% similarity. According to computer analyses of available sequencing data, LCRs may constitute >5% of the human genome. Through the process of non-allelic homologous recombination using paralogous genomic segments as substrates, LCRs have been shown to facilitate meiotic DNA rearrangements associated with disease traits, referred to as genomic disorders. In addition, this LCR-based complex genome architecture appears to play a major role in both primate karyotype evolution and human tumorigenesis. PMID- 12076677 TI - Heart development: learning from mistakes. AB - Congenital heart disease in humans results from abnormal morphogenesis of the embryonic cardiovascular system. The characterization of mutations affecting cardiovascular development in animal models ranging from flies to mice has identified many of the key signaling molecules and transcriptional regulators of heart formation. Many of these molecules are also mutated in familial forms of human congenital heart disease. Through the use of animal models combined with analysis of human pedigrees, a molecular framework that controls formation of the vertebrate heart is beginning to emerge. PMID- 12076676 TI - Smooth, rough and upside-down neocortical development. AB - Lissencephaly, which means 'smooth cortex', is caused by defective neuronal migration during development of the cerebral cortex and has devastating clinical consequences. 'Classical' lissencephaly seems to reflect mutations in regulators of the microtubule cytoskeleton, whereas 'cobblestone' lissencephaly is caused by mutations in genes needed for the integrity of the basal lamina of the central nervous system. Reelin, which is mutated in a third type of lissencephaly, may represent a unifying link because it encodes an extracellular protein that regulates neuronal migration and may also regulate the microtubule cytoskeleton. PMID- 12076678 TI - Hereditary forms of intrahepatic cholestasis. AB - Several genes that are mutated in hereditary forms of intrahepatic cholestasis have been identified or mapped, providing new insights into the process of enterohepatic bile acid circulation in health and disease and new tools with which to study this process. Murine models of several of these disorders have been generated. Unanticipated genetic heterogeneity has been identified. PMID- 12076679 TI - Continuing to break the sound barrier: genes in hearing. AB - The past year has seen major advances in our understanding of the genes involved in Usher syndrome, as well as the discovery of a myriad of other genes expressed specifically in hair cells. Mouse models continue to be invaluable in illuminating our knowledge of how mutations in genes lead to deafness. The role of mitochondrial genes in the hearing process has also contributed to elucidating the workings of the auditory system. PMID- 12076681 TI - The genetics of aging. AB - Once thought to be an extremely complex conundrum of weak genetic and environmental effects, exceptional longevity is beginning to yield genetic findings. Numerous lower organism and mammalian models demonstrate genetic mutations that increase life-span markedly. These variations, some of them evolutionarily conserved, inform us about biochemical pathways that significantly impact upon longevity. Centenarian studies have also proven useful as they are a cohort that, relative to younger age groups, lacks genotypes linked to age related lethal diseases and premature mortality. Pedigree studies have demonstrated a significant familial component to the ability to survive to extreme old age and a recent study demonstrates a locus on chromosome 4 linked to exceptional longevity indicating the likely existence of at least one longevity enabling gene in humans. Thus, a number of laboratories are making substantial and exciting strides in the understanding of the genetics of aging and longevity which should lead to the discovery of genes and ultimately drugs that slow down the aging process and facilitate people's ability to delay and perhaps escape age associated diseases. PMID- 12076680 TI - Muscular dystrophies involving the dystrophin-glycoprotein complex: an overview of current mouse models. AB - The dystrophin-glycoprotein complex (DGC) is a multisubunit complex that connects the cytoskeleton of a muscle fiber to its surrounding extracellular matrix. Mutations in the DGC disrupt the complex and lead to muscular dystrophy. There are a few naturally occurring animal models of DGC-associated muscular dystrophy (e.g. the dystrophin-deficient mdx mouse, dystrophic golden retriever dog, HFMD cat and the delta-sarcoglycan-deficient BIO 14.6 cardiomyopathic hamster) that share common genetic protein abnormalities similar to those of the human disease. However, the naturally occurring animal models only partially resemble human disease. In addition, no naturally occurring mouse models associated with loss of other DGC components are available. This has encouraged the generation of genetically engineered mouse models for DGC-linked muscular dystrophy. Not only have analyses of these mice led to a significant improvement in our understanding of the pathogenetic mechanisms for the development of muscular dystrophy, but they will also be immensely valuable tools for the development of novel therapeutic approaches for these incapacitating diseases. PMID- 12076682 TI - Determination of lopinavir and nevirapine by high-performance liquid chromatography after solid-phase extraction: application for the assessment of their transplacental passage at delivery. AB - An adaptation of the HPLC method previously described for the simultaneous assay of amprenavir, ritonavir, indinavir, saquinavir, nelfinavir and efavirenz after solid-phase extraction is proposed here for the separate analysis of the newer PI lopinavir (LPV) and the NNRTI nevirapine (NVP). After viral inactivation by heat (60 degrees C for 60 min), plasma (600 microl), with clozapine added as internal standard, is diluted 1+1 with phosphate buffer pH 7 and subjected to a solid phase extraction on a C(18) cartridge. Matrix components are eliminated with 2 x 500 microl of a solution of 0.1% H(3)PO(4) neutralised with NaOH to pH 7. LPV and NVP are eluted with 3 x 500 microl MeOH. The resulting eluate is evaporated under nitrogen at room temperature and is reconstituted in 100 microl MeOH 50%. A 40 microl volume is injected onto a Nucleosil 100, 5 microm C(18) AB column. LPV and NVP are analysed separately using a gradient elution program with solvents constituted of MeCN and phosphate buffer adjusted to pH 5.07 and containing 0.02% sodium heptanesulfonate. LPV and NVP are detected by UV at 201 and 282 nm, respectively. The calibration curves are linear up to 10 microg/ml. The mean absolute recovery of LPV and NVP is 91% and 88%, respectively. The method is precise with mean inter-day C.V.s within 2.1-6.6% and 0.9-1.7% for LPV and NVP, and accurate (range of inter-day deviations -1.1 to +2.4%, and -1.9 to +0.8%, for LPV and NVP, respectively). The method has been validated and is currently applied to the monitoring of LPV and NVP in HIV patients, and has been notably applied in a study aimed at assessing the extent of transplacental passage of nevirapine and PIs, notably lopinavir, at the time of delivery in pregnant HIV infected women. PMID- 12076683 TI - Measurement of amoxicillin in plasma and gastric samples using high-performance liquid chromatography with fluorimetric detection. AB - A rapid, selective and sensitive HPLC assay has been developed for the routine analysis of amoxicillin in rat plasma, gastric juice aspirate and gastric tissue which is applicable to low concentrations of amoxicillin (<1 microg mL(-1)) or small sample volumes. Amoxicillin was converted, via an internal rearrangement, to form a fluorescent product which was subsequently recovered using liquid liquid extraction. A Kromasil ODS 3 microm (150 x 3.2 mm I.D.) column was maintained at 40 degrees C and used with a mobile phase consisting of methanol water (55:45, v/v). Fluorimetric detection was at an lambda(ex) of 365 nm and an lambda(em) of 445 nm. The limits of quantitation for amoxicillin were 0.1 microg mL(-1) for gastric juice aspirate (500 microL), 0.5 microg mL(-1) for plasma (50 microL) and 0.075 microg g(-1) for gastric tissue (250 mg). The method was linear up to at least 15 microg mL(-1) in gastric juice aspirate, up to 200 microg mL( 1) in plasma and up to 100 microg g(-1) in gastric tissue, with inter- and intra day RSDs being less than 19%. The assay has been applied to the measurement of amoxicillin in rat plasma, gastric juice aspirate and gastric tissue for pharmacokinetic studies in individual rats. PMID- 12076684 TI - 7,7,8,8-Tetracyanoquinodimethane as a new derivatization reagent for high performance liquid chromatography and thin-layer chromatography: rapid screening of plasma for some antidepressants. AB - Using 7,7,8,8-tetracyanoquinodimethane (TCNQ) as a new derivatization reagent for HPLC and TLC, novel methods are described to detect secondary amine-bearing antidepressants (paroxetine, desipramine, fluoxetine, nortriptyline, maprotiline). The HPLC method is sensitive enough to detect these drugs in plasma at therapeutic levels whereas the latter has potential to detect them in overdose or forensic cases. The methods are based on purple chromogens formed by the displacement reaction of the drugs with TCNQ. The resulting chromogens are directly separated by either reversed-phase HPLC on a C(18) column or TLC on silicagel plates. For HPLC, acetonitrile-water (60:40) was used as mobile phase, with detection at 567 nm and separation in 40 min. For TLC, three developing solvent systems were used. By HPLC, 36 ng ml(-1) spiked plasma concentration of the drugs gave easily detectable signals whereas by TLC, detection limits varied mostly between 240 and 480 ng ml(-1). The HPLC method was applied to real plasma samples. The methods described are simple and very selective; some metabolites of these antidepressants and a vast number of drugs do not interfere with detection. PMID- 12076685 TI - High-performance liquid chromatographic determination of 2-hydroxypropyl-gamma cyclodextrin in different biological fluids based on cyclodextrin enhanced fluorescence. AB - A high-performance size exclusion chromatographic method with analyte enhanced fluorescence detection is described for the analysis of 2-hydroxypropyl-gamma cyclodextrin (HPGCD) in different biological fluids. The principle of detection was the in situ complexation of 8-anilinonaphthalene-1-sulfonic acid (ANS) by HPGCD. When HPGCD eluted from the column the increased fluorescence was measured at excitation and emission wavelengths of 270 and 512 nm, respectively. Solid phase extraction cleanup and concentration of samples resulted in higher than 78% recovery of HPGCD for each of the studied biological fluids. Some important details of the method development as well as the validation of the method for rabbit plasma, rabbit aqueous humour, monkey plasma and monkey urine are given. The limits of quantification varied between 1 and 10 nmol/ml (correspond to 1.5 15 microg/ml) depending on the biological matrix used. The method was successfully adapted in another laboratory proving that HPGCD had not absorbed into aqueous humour and plasma after topical application of HPGCD containing eye drop in rabbits. PMID- 12076686 TI - Liquid chromatographic determination of triethylenetetramine in human and rabbit sera based on intramolecular excimer-forming fluorescence derivatization. AB - A highly selective and simple fluorimetric liquid chromatographic method for the determination of triethylenetetramine (TETA), a therapeutic drug for Wilson's disease, in human and rabbit sera is described. This method is based on intramolecular excimer-forming fluorescence derivatization, which allows spectrofluorometric discrimination of polyamino compounds from monoamino species, followed by liquid chromatography. TETA and 1,6-hexanediamine (internal standard) were converted to the corresponding excimer-forming derivatives with a pyrene reagent, 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester. The derivatives were separated within 20 min on a reversed-phase column using isocratic elution and detected spectofluorometrically at 480 nm with excitation at 345 nm. This method was successfully applied to the monitoring of TETA in human and rabbit sera with a simple pretreatment. The detection limit for TETA in serum was 18 ng/ml (0.13 nmol/ml) corresponding to 0.2 pmol on column at a signal-to-noise ratio of 3. PMID- 12076687 TI - Validated method for quantitation of biomarkers for benzene and its alkylated analogues in urine. AB - A validated gas chromatography-mass spectrometric method for the analysis of the metabolites of benzene and its alkylated analogues in urine is reported. A number of metabolites, as required by authorities for biomonitoring of industrial exposure to aromatic vapour, were analysed simultaneously with preservation of quantitative information concerning positional isomers. The use of this method replaces a combination of analytical methods required for the analysis of all these metabolites. Urine samples were subjected to acidic deconjugation followed by a derivatization step. Phenol, ortho-, meta-, para-cresol, mandelic acid, and ortho-, meta-, para-methylhippuric acid were analysed as their corresponding ethoxycarbonyl derivatives, with single ion monitoring. The mass-to-charge ratios (m/z) of the ions used for quantitation by single ion monitoring of the metabolites were: phenol, 94 m/z; cresols, 108 m/z; mandelic acid, 206 m/z; hippuric acid, 105 m/z; methylhippuric acids, 119 m/z. The mass-to-charge ratios for the internal standards were: [(2)H(6)]phenol, 99 m/z; p-chlorophenol, 128 m/z and 3-chloro-4-hydroxyphenyl acetic acid, 214 m/z. The limits of detection for phenol and the cresols were below 0.4 micromol/l and below 0.05 micromol/l for mandelic acid and the hippuric acids. Within-run precision for mandelic acid was 6.2%, for hippuric acid was 7.32% and was below 5% for the rest of the analytes. PMID- 12076688 TI - Assessment of sirolimus concentrations in whole blood by high-performance liquid chromatography with ultraviolet detection. AB - A novel, reversed-phase high-performance liquid chromatographic (HPLC) method is described for the analysis of sirolimus (SRL) in whole blood. The samples were purified by precipitating blood matrix with zinc sulfate, SRL was then extracted with acetone followed by solid-phase extraction. The method was linear over a range of 1-100 ng/ml and the lower limit of quantification was 2.5 ng/ml. The coefficient of variation (within day) was below 8.0% for the lowest SRL concentration. The day-to-day coefficient of variation was below 6.6%. The assay did not show interference peaks with immunosuppressive drugs commonly given to transplant patients. With the simplified extraction procedure described, 60 samples (including controls and calibration curve) can be quantified in a day. The sensitivity and rapidity of this analytical procedure makes it useful for routine therapeutic monitoring of SRL. PMID- 12076689 TI - Development and validation of a high-performance liquid chromatography-mass spectrometry assay for the determination of artemether and its metabolite dihydroartemisinin in human plasma. AB - A sensitive and selective method is described for the determination of artemether and its active dihydroartemisinin metabolite in human plasma using artemisinin as internal standard. The method consists of a liquid-liquid extraction with subsequent evaporation of the supernatant to dryness followed by the analysis of the reconstituted sample by liquid chromatography-mass spectrometry (LC-MS) in single ion monitoring mode using atmospheric pressure chemical ionization (APCI) as an interface. Chromatography was performed on a C(18) reversed-phase column using acetonitrile-glacial acetic acid 0.1% (66:34) as a mobile phase. The method was fully validated over a concentration range of 5-200 ng/ml using 0.5 ml of human plasma per assay. Stability assessment was also included. The method was applied to the quantification of artemether and its metabolite in human plasma of healthy volunteers participating in pharmacokinetic drug-drug interaction studies. PMID- 12076690 TI - Simultaneous quantification of prostaglandins, isoprostane and thromboxane in cell-cultured medium using gas chromatography-mass spectrometry. AB - We have developed a simultaneous quantification method for prostaglandin (PG) E(2), PGD(2), PGF(2 alpha), 8-epi-PGF(2 alpha), 6-keto-PGF(1 alpha) and thromboxane (TX) B(2). Using [3,3,4,4-(2)H(4)]PGE(2), [3,3,4,4-(2)H(4)]PGD(2), [3,3,4,4-(2)H(4)]8-epi-PGF(2 alpha), [3,3,4,4-(2)H(4)]PGF(2 alpha), [3,3,4,4 (2)H(4)]6-keto-PGF(1 alpha) and [18,18,19,19-(2)H(4)]TXB(2) as internal standards (I.S.), the eicosanoids and their I.S. were simultaneously extracted by solid phase extraction from cell-cultured medium, derivatized to methyl ester/methoxim/tert.-butyldimethylsilyl ether derivatives and analyzed using gas chromatography-mass spectrometry in the selected ion monitoring mode. The accuracy for the added eicosanoids ranged from 92 to 113%, and coefficients of variation ranged from 0.1 to 12.2%. Increased eicosanoids in RAW264.7 and U937 cells stimulated by lipopolysaccharide were suppressed by NS-398 and indometacin. This simultaneous quantification method can be applied routinely for assaying eicosanoids in vitro. PMID- 12076691 TI - Linear regression for calibration lines revisited: weighting schemes for bioanalytical methods. AB - When the assumption of homoscedasticity is not met for analytical data, a simple and effective way to counteract the greater influence of the greater concentrations on the fitted regression line is to use weighted least squares linear regression (WLSLR). The purpose of the present paper is to stress the relevance of weighting schemes for linear regression analysis and to show how this approach can be useful in the bioanalytical field. The steps to be taken in the study of the linear calibration approach are described. The application of weighting schemes was shown by using a high-performance liquid chromatography method for the determination of lamotrigine in biological fluids as a practical example. By using the WLSLR, the accuracy of the analytical method was improved at the lower end of the calibration curve. Bioanalytical methods data analysis was improved by using the WLSLR procedure. PMID- 12076692 TI - Simple liquid chromatographic method for the determination of uracil and dihydrouracil plasma levels: a potential pretreatment predictor of 5-fluorouracil toxicity. AB - 5-Fluorouracil (5-FU) is a commonly used anti-cancer drug with notable activity in clinical practice, yet it causes significant unpredictable and often serious toxicity. Both 5-FU and uracil (U) are catabolised by dihydropyrimidine dehydrogenase (DPD) to form dihydrofluorouracil (FUH(2)) and dihydrouracil (UH(2)), respectively. A means of predicting toxicity before treatment would be more valuable. Variations in dihydropyrimidine dehydrogenase (DPD) activity between patients are at least partly responsible for variable toxicity. Measurement of the UH(2) to U ratio may be a measure of pyrimidine catabolism and thus be utilised to predict subsequent toxicity. We have developed an efficient extraction and detection method using HPLC for the simultaneous measurement of UH(2) and U in plasma. A single C(18) Spherisorb ODS2 (25 cm) column using isocratic elution was utilised. U, UH(2) and the internal standard 4-chlorouracil were detected at wavelengths of 257, 220, and 268 nm, respectively. The chromatographic run time was 45 min which is half that of other methods. The detection limit was 0.02 microM for U and 0.1 microM for UH(2) using only 0.5 ml of plasma for both compounds. The basal plasma concentrations of U and UH(2) in 23 individuals ranged from 0.025 to 0.27 microM and 0.4-1.7 microM, respectively. This simple method may permit the assessment of pyrimidine catabolism, and therefore allow prediction of the toxicities associated with the use of fluorinated pyrimidines. PMID- 12076693 TI - Measurement of 4-hydroxynonenal in small volume blood plasma samples: modification of a gas chromatographic-mass spectrometric method for clinical settings. AB - 4-Hydroxynon-2-enal (4-HNE) is one of the major aldehydic products of lipid peroxidation (LPO) and is involved in a number of pathophysiological processes. Since LPO products are useful indicators for oxidative stress in vivo, a number of detection methods for LPO products in biological tissues were developed. However, none of these methods is presently used in clinical settings. In order to introduce LPO products as biomarkers in clinical studies a suitable GC-MS method for 4-HNE detection was adapted to meet clinical requirements. As one result, the minimal sample volume could be decreased to 50 microl of plasma so that the method might even be suitable for pediatric purposes. The best internal standard (I.S.) for 4-HNE detection by GC-MS 9,9,9-D(3)-4-hydroxynon-2-enal was introduced by van Kuijk et al. [Anal. Biochem., 224 (1995) 420]. However, because of its limited availability, benzaldehyde-ring-d(5), 4-hydroxybenzaldehyde, and 2,5-dihydroxybenzaldehyde were tested to find an alternative. Out of these three, 4-hydroxybenzaldehyde was shown to serve best as I.S. To examine the applicability of the adapted method, tests on the stability of 4-HNE in samples during storage were carried out. It was shown that plasma samples need to be stored at -80 degrees C or less to avoid greater loss of 4-HNE. Samples with 4 HNE concentrations close to the physiological level were shown to be stable over 22 months at -80 degrees C. The introduction of a new and easily available I.S., reduction of the sample volume, and information about sample stability provided by this study facilitate 4-HNE determination in most clinical settings. PMID- 12076694 TI - The aggressive nature of the odontogenic keratocyst: is it a benign cystic neoplasm? Part 2. Proliferation and genetic studies. AB - Immunocytochemical studies of the expression of PCNA, Ki67 and p53 protein have been done by different groups on sporadic keratocysts (OKCs) and OKCs associated with the naevoid basal cell carcinoma syndrome (NBCCS). These 'markers' have in common that they are all expressed in actively proliferating cells, particularly in neoplasms. The findings were compared with their expression in dentigerous and radicular cysts. While there was some variability in the reported results, probably because of technical inconsistencies and the use of different antibodies, a definite trend emerged. In general PCNA, Ki67 and p53 positivity occurred more frequently and more intensely in the OKCs, and in the syndrome related more than the solitary, compared with the other cyst types. In the OKCs the positivity was expressed mostly in the suprabasal layers of epithelium whereas in the other cysts types it was mainly in the basal layer that positivity was observed. Other studies showed that the gene for the NBCCS (PTCH), a tumour suppressor gene, mapped to chromosome 9q22.3. PTCH gene mutation has been shown to be an important step in the pathogenesis of the OKC and was thought to have a role in the development of the sporadic as well as the syndrome-related OKCs. The 'two-hits' hypothesis was invoked in support of the view that syndrome-related basal cell carcinomas (BCCs) and OKCs probably arise from precursor cells that contain an inherited 'first hit'. Only a single mutation was then required in the somatic cell to cause homozygous inactivation and neoplastic progression. Sporadic OKCs might arise from susceptible cells in which two somatic mutations or 'hits' have occurred, one of which manifests as allelic loss. The loss of tumour suppressor genes supports the view that the OKC is a benign neoplasm. PMID- 12076695 TI - Tenascin and beta 6 integrin are overexpressed in floor of mouth in situ carcinomas and invasive squamous cell carcinomas. AB - Floor of the mouth squamous cell carcinomas exhibit many characteristics that suggest they represent a distinct biological subset within head and neck tumors. The features of preinvasive lateral intraepithelial spread, high rate of conversion of intraepithelial neoplasia to invasive carcinoma, and high incidence of occult metastases, suggest the importance of motility-associated proteins in the pathogenesis of these lesions. Two such proteins, tenascin and beta 6 integrin, are generally overexpressed in squamous carcinomas, and may play a central role in the invasive process of floor of the mouth lesions. The purpose of this study was to evaluate in situ and invasive squamous cell carcinomas from the floor of the mouth for the expression of tenascin and beta 6 integrin. Twenty lesions each of floor of the mouth in situ carcinomas and squamous cell carcinomas, and 10 normal controls were stained for tenascin and beta 6 using a standard immunohistochemical protocol for formalin-fixed specimens. Sections were assessed for staining intensity, pattern, and co-localization. Tenascin was highly expressed at the keratinocyte-connective tissue interface of both in situ and invasive carcinomas. beta 6 was expressed in basal keratinocytes of all in situ and invasive lesions, but was not evident in any of the control epithelia. There was no significant difference in staining of in situ and invasive carcinomas, but there was a significant difference in staining between these lesions and controls. Staining was colocalized in serial sections, supporting a receptor-ligand relationship. Both tenascin and beta 6 were weakly expressed in dysplastic areas adjacent to carcinomas suggesting that changes in the expression of these proteins occurs prior to the invasive phenotype. We conclude that tenascin and beta 6 are overexpressed in in situ and invasive floor of the mouth carcinomas, but that transgression of the basement membrane by neoplastic epithelial cells requires additional changes to the keratinocyte molecular profile. PMID- 12076696 TI - A comparative study of the efficacy and safety of fluconazole oral suspension and amphotericin B oral suspension in cancer patients with mucositis. AB - This randomized study compared the efficacy and safety of fluconazole suspension with that of amphotericin B suspension in patients with head and neck cancer who were suffering from candidiasis during cancer treatment with radiotherapy and/or chemotherapy. A total of 123 evaluable patients received 50 mg fluconazole once daily and 120 evaluable patients received 0.5 g amphotericin B thrice daily for 7 14 days depending on clinical response. A positive culture result was obtained in 121 of 264 (46%) patients; Candida albicans was most common. At the end of treatment, fluconazole and amphotericin B were equivalent (CI(90) of -10.7 to +14.9) in terms of clinical cure and improvement, but the rate of mycologic cure was higher for fluconazole (48%) than amphotericin B (35%). The incidence of adverse events was 39% for fluconazole and 44% for amphotericin B. Fluconazole suspension appeared effective and safe. PMID- 12076697 TI - Analysis of fluorescence in oral squamous cell carcinoma. AB - This study was carried out to examine the spectral properties of the red fluorescence emitted from oral squamous cell carcinoma (SCC). Fluorescent samples obtained from oral cancers induced in hamsters, human oral SCCs, and the medium from cultured oral cancer cell lines were analyzed with a spectrofluorometer with excitation at 404 nm. The spectral profile of the experimentally induced cancers changed with increasing malignancy: peaks at 634 and 672 nm increased and peaks at 520 and 582 nm decreased. A reduction in fluorescence intensity at 582 nm and a rise of intensity at 634 nm were commonly observed in the experimental, clinical, and cell line samples, and the ratio of the fluorescence intensity at 582 nm over that at 634 nm was consistent in all samples. These results suggested that the red fluorescence was emitted by porphyrin, which we believe to be produced by oral SCCs and to accumulate inside or on the surface of cancer tissues in greater amounts with progressing malignancy. PMID- 12076698 TI - Effect of herpes simplex virus type-1 on growth of oral cancer in an immunocompetent, orthotopic mouse model. AB - Herpes simplex virus type-1 has been proposed as an agent for the treatment of oral cancer. Experiments were designed to test its effectiveness in an animal model that had a high level of similarity to the human disease. The mouse oral cancer cell line, AT-84, was implanted at an orthotopic site--the base of the tongue--into syngeneic, immunocompetent C3H mice. As expected, tumors invaded the musculature of the tongue, eroded the mandible, and metastasized to the lungs. To obtain a suitable strain of HSV-1 for therapy we screened 17 fresh clinical isolates and selected one that grew to a high titer in vitro. The mouse tumors were then treated by injection of HSV-1 at a titer of 10(9) plaque-forming units/milliliter. To prolong the anti-tumor effect some mice were also given cyclophosphamide, hydrocortisone, or a second injection of virus. To find the importance of bystander killing of tumor cells, some mice were given virus with ganciclovir. A reduction in tumor volume for a limited period was seen after treatment by HSV-1, and was increased by a second injection of virus or by the administration of cyclophosphamide. Ganciclovir negated the anti-tumor effect. Virus was detectable in the tumors for up to 7 days, and loss of virus coincided with the time at which growth of tumors resumed. The mortality of the mice varied up to around 50%. It appears that (1) a non-attenuated strain of HSV-1 can inhibit the growth of an aggressive malignant oral tumor, but only to a limited extent and (2) inhibition depends on the ability of the virus to replicate in the tumor. It is suggested that mutations in the virus will be necessary to prevent mortality, but must be designed carefully so as not to reduce the virulence of the virus. PMID- 12076700 TI - Hyalinizing clear cell carcinoma of salivary glands: a study of extracellular matrix. AB - Hyalinizing clear cell carcinoma (HCCC) is a recently described low-grade carcinoma of the salivary glands presenting two main histological features: clear neoplastic cells and prominent hyalinized stroma. Our aim was to characterize the composition of the hyalinized stroma in three cases of HCCC. An immuno histochemical study using laminin and type I, III and IV collagens, fibronectin and tenascin antibodies was performed. In one case, electron microscopy study was also done. Collagen I and fibronectin were always present in tumoral stroma. Collagen III was documented in two cases and tenascin at the invasive front of the neoplasm. Collagen IV and laminin were found around neoplastic cells but not at the interstitial stroma. Foci of basal lamina reduplication and large deposits of long space collagen were observed in EM. This study demonstrates that the hyalinized stroma of HCCC is not formed by the accumulation of basement membrane type material since it is mostly composed by collagen I and fibronectin. PMID- 12076699 TI - Genetic alterations in squamous cell carcinomas of the hypopharynx with correlations to clinicopathological features. AB - The objective of this study is to describe the molecular alterations in carcinomas in one specific location of the head and neck, the hypopharynx. Thirty seven hypopharyngeal squamous cell carcinomas were studied. The DNA from tumour and healthy tissue was evaluated for amplification of the 11q13 region and of the MYC and ERBB1 oncogenes, for integration of the Human Papillomavirus (HPV), and for loss of heterozygosity (LOH) at p53 and NAT2 loci. The most common alteration was the amplification of the 11q13 region (78% of the cases), followed by LOH at p53 locus (70%). MYC amplification was found in 19% of the cases, ERBB1 amplification in 29%, LOH at NAT2 locus in 25%, and integration of the HPV in 29%. 11q13 amplification was related with nodal metastases and higher tumour recurrence rates. These findings confirm that 11q13 amplification is one of the most frequent genetic alterations in hypopharyngeal squamous cell carcinomas, and that it may have prognostic significance in these tumours. PMID- 12076701 TI - Assessment of dynamic MRI of Warthin's tumors arising as multiple lesions in the parotid glands. AB - We examined three patients with multiple synchronous Warthin's tumors in the bilateral parotid and described the value of using dynamic MRI. The time course of the contrast index (CI curves) was calculated from a dynamic series. Warthin's tumors showed intermediate signal intensity on T1WI, heterogeneous high and intermediate signal intensity on T2WI and a slight enhancement on Gd-T1WI. Warthin's tumors of CI curves showed specific findings. CI curves in each lesion showed the same pattern. It was difficult to diagnose masses as Warthin's tumors using only MR images on T1, T2 and Gd-T1WI. Dynamic MRI can distinguish Warthin's tumors from other possible tumors except for oncocytoma. Therefore, the use of dynamic MRI is recommended as a diagnostic method for Warthin's tumors in multiple synchronous lesions of the parotid gland. PMID- 12076702 TI - Views of oral cancer prevention and early detection: Maryland physicians. AB - The purpose of this study was to obtain in-depth information on Maryland physicians' knowledge, opinions and practices about oral cancer examinations. The qualitative descriptive study used one focus group conducted in a conference facility and nine one-on-one interviews at private medical offices. A criterion purposeful sampling was used for selection of participants. Generally, we found low awareness of, and surprise about, Maryland's high oral cancer mortality rates. Physicians were not surprised that they detect more lesions than dentists, although most physicians did not provide oral cancer examinations on a routine basis. Physicians were interested in attending continuing medical education (CME) courses on oral cancer prevention and early detection but only if worked into other CME programs on cancer. They were very interested in having hands-on training on performing an oral cancer examination. These findings will be used to implement educational interventions for Maryland physicians to help increase early detection of oral cancers. PMID- 12076704 TI - A novel, de novo germline TP53 mutation in a rare presentation of the Li-Fraumeni syndrome in the maxilla. AB - We undertook the genetic analysis of a classic Li-Fraumeni syndrome (LFS) family with clustering of primary tumours including two maxillary sarcomas, a rare LFS site of tumour occurrence. Our aim was to investigate the presence of a specific type of TP53 mutation that could be associated with this unusual predilection of site for cancer occurrence. Mutational screening of the coding region of TP53 revealed an A>T transversion in codon 144 of exon 5 (CAG>CTG, Gln>Leu) in the germline of one of the three affected members, with loss of heterozygosity (LOH) in the tumour tissue. All other affected members were negative for germline or somatic TP53 mutations. TP53 immunohistochemistry was uninformative. The mutation we report is a de novo constitutional TP53 mutation that has not been previously described in the literature. It could explain the more burdened phenotype of the affected patient (died at 21 months). Alternative mechanisms to explain the overall family phenotype are discussed. PMID- 12076703 TI - Evaluation of T-classifications of upper gingival and hard palate carcinomas--a proposition for new criterion of T4. AB - Most carcinomas of upper gingiva and hard palate are classified as T4 stage on the basis of the UICC criteria, since they easily invade the underlying bone tissue. We classified 43 patients with squamous cell carcinoma of the upper gingiva in terms of three criteria: (1) the original T-classification by UICC, (2) the classification by the Japan Society for Head and Neck Cancer (JSHNC), and (3) a new classification in which the maxillary sinus or nasal floor is used as the defining borderline for T4 (MSF classification). Our study demonstrated that the new classification was superior with regard to distribution of patients by T stage, correlation with prognosis and choice of treatment method. PMID- 12076705 TI - Oral yeast carriage correlates with presence of oral epithelial dysplasia. AB - Previous studies have suggested a link between the presence of Candida albicans and the development of oral squamous cell carcinoma (OSCC). The aim of the present study was to assess the presence and level of colonisation of oral yeast in patients undergoing an incisional oral mucosal biopsy in order to assess whether the amount of oral yeast present correlated with the presence and degree of oral epithelial dysplastic or neoplastic change. Two hundred and twenty-three patients who were undergoing an incisional biopsy for the diagnosis of an oral mucosal lesion were enrolled in this study. Mouth swills were obtained from each patient for the presence and amount of oral yeast present. Some of the patients (44.6%) had a histopathological diagnosis of either oral epithelial dysplasia (OED) or OSCC and the frequency of oral yeast carriage was significantly greater (P<0.001) in these patients than those without histopathologically detected dysplastic or neoplastic oral lesions. Furthermore, significantly (P<0.001) more patients with OED or OSCC had a higher number of yeast (over 1000 cfu/ml) in their oral cavity than patients without any evidence of epithelial dysplasia or neoplasia histopathologically. The degree of epithelial dysplasia present in these patients also correlated with higher amounts of yeast in the oral cavity (P=0.017). The results of the present study reveal that there is an interaction between oral carriage of yeast and oral epithelial dysplasia, however it remains unclear how yeast infection influences the development and progression of dysplasia. PMID- 12076707 TI - Intraduct papilloma of the palate. Report of a case. AB - Salivary gland papillomas are rare tumours arising from ductal epithelium. Intraduct papillomas are the most rare of all duct papillomas. Only four intraduct papillomas have been described in a review of nearly 3100 epithelial salivary tumours. Intraduct papillomas are located almost exclusively in the excretory ducts of the minor salivary glands. However, also the major salivary glands may be affected. Microscopically, the tumour consists of fibrovascular papillae covered by a columnar or cuboidal epithelium. The authors describe an intraduct papilloma of the palate in a 74-year-old woman. The excision of the lesion was curative. PMID- 12076706 TI - Importance of tumour thickness measurement in prognosis of tongue cancer. AB - Eighty-one patients who underwent surgery for cancer of the tongue were retrospectively studied to evaluate the influence on survival of some clinical and pathologic parameters. These parameters and data on the patient's current status were gathered by the study of tissue sections, using haematoxylin-eosin staining, and from medical records. The 5-year survival rate was 68.5%. Univariate analysis showed that the parameters influencing survival were: T (P<0.01), pathologic T (P<0.01), N (P<0.05), pathologic N (P<0.05), extracapsular nodal spread (P<0.05), locoregional recurrence (P<0.01), and tumour thickness (P<0.05). Multivariate analysis showed that tumour thickness had the greatest influence on survival. Patients with tumour thickness of < or = 3 mm had a 5-year survival of 85.7%, significantly greater (P<0.05) than the rates of 58.3 and 57% for patients with tumour thickness of 4-7 mm and >7 mm, respectively. Wider studies are required to unify criteria for the measurement of this important prognostic parameter. PMID- 12076708 TI - Primary high-grade mucoepidermoid carcinoma of the minor salivary glands with cutaneous metastases at diagnosis. AB - Mucoepidermoid carcinoma (MEC) represents 15% of all salivary glands malignancies. Metastatic disease at diagnosis is observed in less than 5% of the cases. The lung is the most commonly involved site. This is the first reported case of high-grade MEC of the salivary gland with skin metastases at diagnosis. This feature was associated with a chemoresistant and aggressive behaviour. Differential diagnosis between metastatic MEC and primary skin MEC is essential for therapeutic management and prognosis. PMID- 12076710 TI - Fas-ligand and perforin expression in infiltrating cytotoxic T lymphocytes in the liver of chronic hepatitis C. AB - Cytotoxic T cells (CTLs) are believed to play an important role in the pathogenesis of chronic hepatitis C based on histological findings of the liver and in vivo experiments. Fas-ligand-Fas and perforin dependent pathways are two major killing systems when CTLs induce their target-cell death. Thus, the present study attempts to determine whether these pathways are activated, and if they are, how they are related in chronic hepatitis C. To investigate the expression of Fas-ligand and perforin, we performed double immunofluorescent staining of liver biopsy specimens from patients with chronic hepatitis C. Fas-ligand and perforin expression was observed in mononuclear cells, and the partial coexistence of the two proteins was observed. Cells expressing both proteins were positive for CD45RO(+) T cells (active T cells), whereas cells expressing perforin were negative for CD68 (macrophages). In the cases which had sustained negative HCV-RNA over 6 months after interferon treatment, Fas-ligand was not expressed, although perforin was slightly detectable. To quantitatively assess the balance of these pathways, hepatic mRNAs of Fas-ligand and perforin were measured by quantitative RT-PCR. The ratio of Fas-ligand-mRNA/perforin-mRNA was significantly correlated with serum alanine aminotransferase (ALT) levels (r=0.913). These results suggest that both pathways are activated in chronic hepatitis C and that Fas-ligand-Fas pathway may be predominant in active inflammation. PMID- 12076711 TI - Association of circulating CD8(+) lymphocytes to a spontaneous and interferon alpha induced clearance of HCV. AB - The amount of copies of HCV-RNA and count of CD8(+) lymphocytes was retrospectively evaluated in 326 patients: sampling was performed in basal condition, during treatment with alpha-IFN (n=232) and post-treatment follow-up, and at the same time points in untreated patients (n=94). In the treated group the difference between CD8(+) lymphocytes in the patients successfully treated (n=65) and those with an unfavourable outcome (n=176) is statistically significant (898+/-172 vs., 440+/-176 CD8(+) lymphocytes per mm(3) P<0.005 ANOVA). Also, in the untreated patients the average count of CD8(+) cells is statistically higher in patients with a favourable outcome (P<0.01 ANOVA). The present data show that the count of CD8(+) lymphocyte is of clinical value in order to predict the outcome of HCV infection and may be used together with the viral load and genotype, already established predictors. PMID- 12076712 TI - Clinical features and viral sequences of various genotypes of hepatitis B virus compared among patients with acute hepatitis B. AB - Clinical manifestations and viral sequences of core promoter and precore/core region were compared among various genotypes of hepatitis B virus (HBV) in 25 patients with acute hepatitis. The genotype in patients with acute hepatitis was distributed differently from that among chronic hepatitis patients in Japan, which are predominantly genotypes B and C. Of 25 patients with acute hepatitis, 14 had genotype A, five genotype B and six genotype C. Serum total bilirubin levels were significantly higher in patients with genotype A than in those with genotype C. Prothrombin time was shorter in patients with genotype B than those with genotype A or C. Total bilirubin was lower in patients with short duration of acute hepatitis. The serum ALT value remained above 1000 IU/l for over 10 days in 79% of patients with genotype A. This prolonged duration of hepatitis in patients with genotype A may contribute to hyperbilirubinemia. Sequence analysis revealed no difference in the number of mutations in precore/core regions among the three genotypes. Although the double mutation, A-T and G-A at 1762 and 1764, respectively, was found in two patients each with genotype A and C, these mutations were not related to the hepatitis B e antibody (HbeAg)/hepatitis B e antibody (HbeAb) phenotype. Two of seven patients with thymidine at 1858 also had a G to A mutation at 1896. Thus, the difference in the genotype little influenced the HBeAg/HBeAb phenotype in acute hepatitis patients. Understanding the viral genotypes in acute HBV infection may be valuable in predicting the clinical course of acute hepatitis B (AHB). PMID- 12076713 TI - Superoxide dismutase and total anti-oxidant levels in various forms of liver diseases. AB - In order to understand the impact of viral hepatitis on anti-oxidant defence system of the body, blood levels of superoxide dismutase (SOD), an enzymatic anti oxidant, and total anti-oxidant (TAO) were evaluated and co-related to etiological viral hepatitis in various forms of liver diseases. A total number of 110 patients including 50 patients with acute viral hepatitis (AVH), 30 patients with chronic active hepatitis (CAH) and 30 patients with cirrhosis of liver were analysed for different hepatitis viral markers and the anti-oxidant levels in their blood. For comparison, blood from 100 healthy persons were also simultaneously tested for anti-oxidant levels. Analysis of results indicated that none of the patients belonging to these three liver diseases had hepatitis A viral (HAV) and hepatitis D viral (HDV) infections. AVH group had mainly hepatitis B viral (HBV), hepatitis C viral (HCV) and hepatitis E viral (HEV) infections, CAH group had B and C infections and cirrhosis group had B, C and E infections. A sizeable number of patients in each group had no markers and were labelled as non-BCE group. On co-relating anti-oxidant levels to viral etiology in these patients, it was observed that in comparison to healthy control group, SOD level was significantly reduced in all the patients irrespective of the viral etiology (P<0.05-0.001). The impact of different viruses on reduction in SOD level was recorded to be the same with no significant difference in SOD level between any two viral infections. On the contrary, TAO level in the majority of patients was found to be comparable with that observed in healthy persons. An appreciable change in SOD level but little impact on TAO level during viral hepatitis may be explained by the possible adaptive rise of some other anti oxidant level in the blood of these patients. PMID- 12076714 TI - Expression of cyclooxygenase 2 and cytosolic phospholipase A(2) in the liver tissue of patients with chronic hepatitis and liver cirrhosis. AB - Cyclooxgenase (COX) and phospholipase A(2) (PLA(2)) are crucial rate-limiting enzymes involved in the conversion of arachidonic acid to prostaglandin H(2), the precursor of various compounds including prostaglandins (PGs), prostacyclin, and thromboxanes in the process of PGs' synthesis. Recent studies have shown increased levels of COX(2) in adjacent cirrhotic tissue of hepatocellular carcinoma. The relationship between the expression of COX(2) or cytosolicPLA(2) (cPLA(2)) and liver fibrosis has not been described previously. We used 45 formalin-fixed, paraffin-embedded liver tissue samples obtained by needle biopsies from patients with chronic hepatitis, consisting of 7 cases of F(0), 10 cases of F(1), 10 cases of F(2), 9 cases of F(3) and 9 cases of liver cirrhosis (LC) according to the New Inuyama Classification of the staging of liver fibrosis. The expression of COX(2) and cPLA(2) was investigated by immunohistochemistry, western blotting and image analysis. The positive signals for COX(2) and cPLA(2) were observed in the cytoplasm of hepatocytes. The signal intensity of COX(2) increased significantly with the progression of liver fibrosis (P<0.001) and no significant difference was observed in the relative amount of cPLA(2) from group F(0) to group LC. According to the New Inuyama Classification of hepatitis activity grading, 45 samples were classified as group A(1) (23 cases), group A(2) (19 cases) and group A(3) (3 cases). No significant differences were found in the relative amount of COX(2) and cPLA(2) between group A(1) and group A(2-3). Significant correlation was observed between the relative amount of COX(2) and hyaluronan (P<0.01). Our findings suggested that COX(2) may be involved in liver fibrogenesis. PMID- 12076715 TI - Relationship between the histological degrees of hepatitis and the postoperative recurrence of hepatocellular carcinoma in patients with hepatitis C. AB - The relationship between the recurrence of hepatocellular carcinoma (HCC) and the degree of inflammation was evaluated in resected livers with the hepatitis C virus (HCV) -associated HCC. Seventy-three patients with HCV-associated HCC who were followed up for more than 2 years were selected for this study. In these cases, the degree of chronic hepatitis in noncancerous regions at the time of surgery was classified according to the New Inuyama Classification as follows, the degree of necroinflammatory activity (Grading) was graded from A0 to A3, and the degree of fibrosis (Staging) was staged on F0-F4. In addition, among these patients, 41 patients who were followed by blood tests every 3 months were divided into two groups (high or low group) according to annual average levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), the platelet counts (Plt), and alpha-fetoprotein (AFP). As a result, cancer-free survival rate was significantly lower in the high-grade group (A3) than in the low-grade group (A1 or 2) (P=0.01). The high ALT (>80 IU) group and the high AFP (>20 mg/ml) group also had significantly worse cancer-free survival rate than the low ALT group and the low AFP group (P=0.04 for ALT, P=0.03 for AFP). A multivariate analysis for the prognostic values revealed the AFP level (P=0.02) and the Grading (P=0.04) were useful as independent prognostic factors concerning recurrence. In conclusion, the degree of inflammatory activity (Grading) is considered to be a useful factor regarding recurrence after liver resection in patients with HCC. Furthermore, the inhibition of inflammation in remnant liver may also contribute to the prevention of recurrence. PMID- 12076716 TI - Survival and local recurrence rates of hepatocellular carcinoma patients treated by transarterial chemolipiodolization with and without embolization. AB - The factors contributing to local recurrence and survival rates of patients with hepatocellular carcinoma (HCC) were analyzed by univariate and multivariate analysis using the Cox proportional hazard model in 356 patients treated by transcatheter arterial chemoembolization (TAE) or transcatheter arterial chemolipiodolization (TAI). Potential predictors of recurrence and survival analyzed included: patient characteristics (gender and age), basal liver disease, tumor characteristics (number of tumors, size of tumor nodule and presence of vascular invasion), Child classification, serum albumin and total bilirubin level, prothrombin time, treatment modality (TAE or TAI) and serum levels of the tumor markers [alpha-fetoprotein (AFP) and des-gamma-carboxy prothrombin (PIVKA II)]. In 259 patients (72.5%) with confirmed complete necrosis of the primary lesion, the local recurrence rate in the TAE group was significantly lower than that of the TAI group (P<0.05). Size of tumor nodule, vascular invasion, treatment modality (TAE or TAI) and serum PIVKA-II level were significant independent risk factors contributing to local recurrence. Serum albumin level, size of tumor nodule, vascular invasion and serum AFP level were significantly independent prognostic factors contributing to survival. When size of tumor nodule was greater than 30 mm in diameter and without vascular invasion, TAE had a survival benefit superior to TAI. PMID- 12076717 TI - Involvement of Smad proteins in TGF-beta and activin A-induced apoptosis and growth inhibition of liver cells. AB - Transforming growth factor (TGF)-beta and activin A inhibit the growth and induce cell death of parenchymal liver cells. Smad proteins have recently been identified as intracellular signaling mediators and modulators of TGF-beta family members. This study assessed the role of Smad proteins during the action of TGF beta and activin A on liver cells using a well-differentiated human hepatoma cell line, Hep3B cells. To study the role of Smad proteins in the anti-proliferative, apoptosis-inducing, activities of TGF-beta and activin A in liver cells, we stably transfected dominant negative Smad2-3SA or Smad3-3SA mutants in Hep3B cells. Transfection of Smad2-3SA or Smad3-3SA abrogated both TGF-beta-induced and activin A-induced growth inhibition and apoptosis of Hep3B cells. Down regulation of Bcl-xL expression by TGF-beta was both Smad2 and Smad3 dependent. We also demonstrate that transfection of Smad7, an intracellular antagonist of Smad signaling, inhibited both TGF-beta- and activin A-induced apoptosis and growth inhibition of these cells. These results suggest that Smad proteins positively and negatively mediate TGF-beta-induced and activin A-induced apoptosis and growth inhibition of liver cells. PMID- 12076718 TI - The usefulness of prevention consolidation therapy of esophageal varices using an argon plasma coagulation technique. AB - BACKGROUND AND STUDY AIM: esophageal variceal ligation (EVL) has been spread widely as a simple and effective method for treating esophageal varices. However, in view of the unacceptably high rate of recurrence associated with this technique, the availability of other supplemental consolidation therapies has been earnestly desired. Under these circumstances, attempts such as endoscopic injection sclerotherapy (EIS) have been made for this particular therapeutic purpose. In the current study, we tested the clinical usefulness and safety of the argon plasma coagulation technique (APC). PATIENTS AND METHODS: 11 patients with imminent signs of esophageal varix rupture in our hospital were subjects in the present study. Before APC was performed, these patients underwent EVL with consequent improvement of esophageal varices from F3 (largest sized varices) to disappear or F1 (straight). RESULTS: the patients were followed up for a mean post-treatment period of 637.4 (SD +/-56.5) days, and no obvious recurrence of varices in the so-called critical area was noted. In addition, no serious complications were observed. CONCLUSION: our current clinical study has demonstrated that APC may be an effective prevention consolidation therapy after EVL without serious complications. PMID- 12076719 TI - Environmental manipulations in rodents and primates. Insights into pharmacology, biochemistry and behaviour. PMID- 12076720 TI - Handling, genetic and housing effects on the mouse stress system, dopamine function, and behavior. AB - This research was designed to examine how early stimulation (i.e., handling), subsequent housing conditions and genetic factors interact to produce adult differences in stress regulation. High-aggressive (NC900) and low-aggressive (NC100) mice were handled for 3 weeks potspartum and were subsequently isolated or grouped until observed as adults in an open field or a dyadic test. In NC100, handling abolished the temporal variations seen in open-field activity among the nonhandled subjects and reduced corticosterone (CORT) activation. In NC900, these two measures were unaffected by handling. Only among handled NC100 did subsequent group rearing further reduce CORT activation. By contrast, handling caused an up regulation of D1 dopamine receptors in both lines, and, in NC100, this effect was increased by group rearing. In a dyadic encounter with another male mouse, subjects of both lines showed handling effects. NC100 froze less rapidly and NC900 attacked more rapidly. This multifactorial design showed that the systemic effects of handling are modulated by genetic background, and that measures of these effects are affected by experience beyond infancy. Our findings also showed that the effects of handling vary when assessed across different physiological systems and across social and nonsocial testing conditions. PMID- 12076721 TI - Neurobehavioral development of two mouse lines commonly used in transgenic studies. AB - The present study was aimed at establishing the differences in the neurodevelopmental profile between two F2 lines derived from two F1 hybrid mouse strains (129 x C57BL/6 and C57BL/6 x SJL). The choice of the given strains was based on the frequent use of these mice in transgenic research. For the neurodevelopment phenotyping, we employed a test battery consisting of 23 somatometric, sensorial and motor tests. Significant variations between the strains were established in different functional domains. Some specific delays in the appearance of developmental landmarks were observed in F2 mice derived from crosses of F1 C57BL/6 x 129, whereas they acquired early developmental functions, such as the righting reflex, sooner than C57BL/6 x SJL-derived mice. C57BL/6 x 129 F2 offspring were spontaneously hypoactive, and their poorer motor performance was confirmed by low performance in the negative geotaxis test. However, there were no differences in the general psychomotor development as shown by the good performance in the homing test in both F2 lines. Both strains were susceptible to the handling procedures used, presenting a similar alteration in the response observed in the homing test as compared to nonhandled control mice. In conclusion, our work highlights the importance of the genetic background for transgenesis experiments and also the need for well-established testing protocols to obtain sufficient information at the first stage of behavioral screening of genetically modified mice. PMID- 12076722 TI - Craving for alcohol in the rat: adjunctive behavior and the lateral hypothalamus. AB - A review of previous results and the new data in this report show clearly that the Falk model of adjunctive behavior is an adequate analogue of human alcoholism and can be applied to induce excessive ethanol consumption. New data on the consumption of sweet flavored ethanol solutions and, especially, sweet alone solutions during brief periods of ethanol withdrawal provide some significant insights concerning the possible physiological basis for cravings in humans. Because voluntary consumption of ethanol is the normal process by which alcoholism develops, a general set of environmental and other experimental conditions that produce behavioral excess; adjunctive behavior, electrical stimulation of the brain, and salt arousal of drinking are discussed in some detail. Neuronal circuits of the lateral hypothalamus are important because some of the cells are chemosensitive and monitor osmolality of the blood and initiate drinking in the normal regulation of body fluids. Alcohol in very small amounts has a direct effect on these cells that also project to lower spinal motor neurons and modulate the level of excitability in spinal reflexes and thereby reactivity to environmental stimulation. Taste and other sensory information from the mouth arrives in presynaptic endings on these same cells by a multitude of indirect multisynaptic pathways. A theoretical model is developed to explain how tactile and taste sensory information and what is initially a nonspecific general state of motor arousal interact together to produce an excessive consumption or craving for ethanol. PMID- 12076723 TI - Postnatal stress of early weaning exacerbates behavioral outcome in prenatal alcohol-exposed juvenile rats. AB - Some of the behavioral deficits caused by prenatal or postnatal alcohol exposure have been demonstrated to be ameliorated by environmental manipulations such as handling or environmental enrichment. This experiment, in contrast, investigated whether behavioral deficits due to prenatal alcohol exposure could be exacerbated by a stressful experience, early weaning. Pregnant dams were given either a liquid diet with 35% of the calories derived from alcohol, a liquid diet without alcohol to control for any effects of the liquid diet administration, or ad libitum food and water. Half of each litter were weaned at 15 days of age (early weaning) and half were weaned at 21 days of age (normally weaned). Offspring were weighed, tested for activity in an open field at 18 days of age, and trained to find a hidden platform in the Morris water maze at 22-24 days of age. Alcohol exposed subjects who were weaned early were more impaired in spatial navigation ability than any other group. Similarly, the combination of early weaning and prenatal alcohol exposure caused the slowest growth. All subjects exposed to alcohol, regardless of weaning condition, had greater latencies to find the platform than those from the two control groups. There was no synergistic effect of alcohol and stress on activity levels, but all early-weaned females were more active than normally weaned females; males did not show this effect. Thus, environmental stressors such as early weaning can compound detrimental symptoms of prenatal alcohol exposure. These results have implications for the understanding of the effects of the environment on neuronal plasticity. PMID- 12076724 TI - Prenatal stress produces deficits in socio-sexual behavior of cycling, but not hormone-primed, Long-Evans rats. AB - Prenatal stress (PNS) alters behavior of adult offspring in novel environments or in social interactions; variable effects of PNS on female reproductive behavior have been reported. Effects of exposure to restraint and lights for 45 min/day on Gestational Days 14-20 were examined on the motor and socio-sexual behavior of adult female offspring. In a novel arena, proestrous PNS rats displayed greater behavioral inhibition as indicated by significantly fewer beam breaks made in the horizontal crossing task compared to that of proestrous non-PNS rats. In a standard mating test, in which females are exposed to males in a relatively small space for a restricted time or number of sexual contacts, PNS females in proestrus were found to have significant decreases in the intensity of lordosis and in the number of solicitation behaviors that they directed towards the male compared to non-PNS rats. In a seminatural mating test, in which females can control the timing of the sexual contacts from the male, PNS females in proestrus engaged in significantly less pacing of their sexual contacts compared to that of the non-PNS females. When additional PNS and non-PNS rats were ovariectomized (ovx) and tested following hormone priming, behavioral differences were abrogated. PNS decreased motor behavior in a novel arena, lordosis intensity, and solicitation behavior in a standard mating paradigm, as well as adaptive, approach-avoidance behavior in a seminatural mating situation of endogenously cycling proestrous rats but not ovx, hormone-primed rats. Thus, hormone priming may override or mask effects of PNS on some aspects of socio-sexual behavior. PMID- 12076725 TI - Mothering begets mothering: the transmission of behavior and its neurobiology across generations. AB - Early experiences exert their effects on adult parental behavior in part by altering the development of neurobiological mechanisms that initiate or support the initiation and sustenance of adult parental behavior. The effects of parental behavior on sensory, perceptual and emotional mechanisms in offspring constitute an experientially based mechanism by which neurobiological factors regulating behavior can be transferred from generation to generation somewhat independently of genetic endowment. PMID- 12076726 TI - Neonatal isolation alters stress hormone and mesolimbic dopamine release in juvenile rats. AB - Rat pups were individually isolated from the mother and nest for 1 h/day from postnatal days (PND) 2 to 9 and tested as juveniles (PND 26-30) compared to nonhandled (NH) controls. In response to 1 h of restraint stress, NH rats increased locomotor activity and dopamine (DA) levels, but neonatally isolated (ISO) rats did not. Both groups had increased plasma corticosterone levels in response to restraint, but corticosterone levels were higher in ISO than in NH. Brain allopregnanolone (3alpha,5alpha-THP) levels also increased in response to stress, but NH and ISO did not differ. Sex of the rats was not a factor for any of the measures except plasma corticosterone levels, where females had higher levels than males. These data indicate that the effects of neonatal isolation persist postweaning and that the effects are most evident in response to stress as opposed to under baseline conditions. PMID- 12076727 TI - Peripheral benzodiazepine receptors reflect trait (early handling) but not state (avoidance learning). AB - Behavioral animal paradigms and experimental neuroendocrinological and neurochemical studies have shown that early environmental manipulations have profound effects on the late response to stress. The aim of the present study was to investigate the interactive effects of environmental manipulation (early handling) and experimentally induced behavioral differences on the peripheral benzodiazepine receptor (PBR) system, which is known to be involved in the response to stressors. Adult early-handled (EH) and nonhandled (NH; control) Wistar rats were placed in a two-way active avoidance/latent inhibition (LI) paradigm, and PBR densities in the adrenal glands, kidneys, and gonads were assessed. In line with previous studies, overall avoidance learning improved in the EH group, and LI was disrupted in the NH group (primarily in males). PBR densities were up-regulated in EH subjects, and more so in females than males. However, PBR densities did not correlate with any of the behavioral measures. These findings strengthen the hypothesis that differences in PBR densities between EH and NH rats are a reflection of trait rather than state, and they suggest that the PBR system is characterized by a highly stressor-specific response. PMID- 12076728 TI - Corticosterone administration to rat pups, but not maternal separation, affects sexual maturation and glucocorticoid receptor immunoreactivity in the testis. AB - Prenatal stress strongly affects sexual dimorphism of male rats. Much less information is instead available on the effects of postnatal stress on sexual maturation during the so-called stress hyporesponsive period (SHRP). For this reason, we compared corticosterone-treated (CS; 10 mg/kg sc, suspended in sesame oil) or maternally separated pups (MS; 5 h/day in the first week of life) with control rats. Control and MS pups also received sesame oil injections. The effects of these procedures on physical development (body weight and eye opening), sexual maturation [anogenital distance, testis weight, 3beta hydroxysteroid dehydrogenase/(Delta5-4) (3betaHSD) isomerase activity and time to testis descent] and glucocorticoid receptor (GR) immunoreactivity in the testis were examined. Corticosterone treatment significantly (P<.05) advanced testis descent and increased testis weight and 3betaHSD activity at puberty. In addition, adult CS rats presented higher levels of GR immunoreactivity in testicular tubules when compared to control and MS rats. No differences were found between control and MS rats. On this basis, we propose that the silencing of adrenocortical function during the SHRP could be finalized to preserve sexual maturation from the influence of glucocorticoid effects. As SHRP is unique to rodents, this phenomenon could be related to their successful reproductive strategy. PMID- 12076729 TI - Maternal corticosterone influences behavior, stress response and corticosteroid receptors in the female rat. AB - In infancy, glucocorticoids have been shown to affect hypothalamus-pituitary adrenal (HPA) axis activity and behavior. Both the activity of the HPA axis and many aspects of behavior exhibit important gender-dependent differences physiologically. In our previous studies, male offspring of hypercorticosteronemic mothers show long-lasting changes of learning as well as adrenocortical activity. In the light of these findings, this study aims to determine the long-term effects of glucocorticoids in the early stages of life in female rats. Corticosterone (200 microg/ml) was added to the drinking water of the dams. Female offspring exhibited lower adrenocortical secretory response to stress, improvement in learning (water maze at 21, 30 and 90 days; active avoidance at 15 months) and reduced fearfulness in anxiogenic situations (dark light test at 1 and 15 months; conditioned suppression of drinking at 3 months; plus maze at 15 months) after weaning, from 21 days up to 15 months of age, but not before. No difference in hippocampal adrenocorticoid receptors was observed. These results, together with previous data on male offspring, show that the outcomes of maternal hypercorticosteronemia on hormonal stress response and behavior are similar in males and females, but the effects on some aspects of the HPA axis activity are gender-dependent. Possible explanations for these differences are discussed. PMID- 12076730 TI - Repeated maternal separation does not alter sucrose-reinforced and open-field behaviors. AB - Repeated separation of rat pups from their mothers has been reported to increase behavioral fearfulness and hypothalamic-pituitary-adrenal (HPA) response to stress. Recently, it was suggested that it might also alter behavioral responses to natural and drug rewards. Here, we studied whether maternal separation (MS) would alter behavioral responses to a sucrose reward. We also tested whether MS would alter behavioral responses in an open-field test using a novel method of analysis [Software for the Exploration of Exploration (SEE)]. Long-Evans rat pups were exposed to either 180 min of MS, 15 min of separation [early handling (EH)] or left undisturbed [nonhandled (NH)] from postnatal day (PND) 3 to 14. The adult male offspring were tested for sucrose solution preference using a two-bottle free-choice test, operant response for sucrose under fixed ratio and progressive ratio (PR) schedules of reinforcement and response to a novel environment (open field test). MS had no effect on sucrose preference or operant responding for sucrose reward. In the open-field test, NH rats showed a brief decrease in locomotor response, but MS rats did not differ from the NH and EH groups in the other behavioral measures. Thus, under the conditions of the present study, MS did not appear to alter reward-related processes and also had a minimal effect on open-field behavior. PMID- 12076731 TI - Effects of maternal separation on brain nociceptin/orphanin FQ peptide levels in male Wistar rats. AB - Environmental manipulation early in life may induce persistent alterations in adult behaviour and physiology. In this study, we investigated the long-term effects of daily maternal separation, Days 1-21, on brain immunoreactive nociceptin/orphanin FQ (ir-N/OFQ) levels in male Wistar rats. The rat pups were separated in litters for 360 min (MS360) or 15 min (H15). Control rats were left undisturbed until weaning. Peptide levels were measured at 10 weeks of age. In the hypothalamus and periaqueductal gray, MS360 induced an increase in ir-N/OFQ levels in comparison with control rats. H15 rats had increased ir-N/OFQ levels in the hypothalamus and the medial prefrontal cortex compared with control animals. The rats were also tested at two occasions in an elevated plus-maze. An increased anxiety-like behaviour was shown in MS360 rats at weaning, whereas a decreased anxiety response was found at 9 weeks of age compared with control rats. The study shows that early life experiences induce long-term effects on behaviour, as well as brain N/OFQ levels. PMID- 12076732 TI - Long-lasting changes in stress-induced corticosterone response and anxiety-like behaviors as a consequence of neonatal maternal separation in Long-Evans rats. AB - Early neonatal environmental factors appear to have powerful and long-lasting influences on an organism's physiology and behavior. Long-Evans male rats separated from their dam for 3 h daily over the first 2 weeks of life (maternally separated, MS rats) when tested as adults exhibit exaggerated behavioral and neuroendocrine responses to stress compared to 15-min separated (handled, H) animals. The purpose of this study was to compare male and female adult rats that were MS, H or were undisturbed (nonhandled, NH) as neonates in anxiety-like behaviors, in the elevated plus-maze, and in response to startle-inducing auditory stimuli. We confirmed that MS males oversecrete corticosterone (CORT; 2.5-5 times) in response to mild handling stress. MS males and females were less likely to explore open arms of the plus-maze. MS males exhibited 35% higher startle amplitudes compared to controls. Furthermore, MS males were more likely to emit ultrasonic vocalizations in response to startle than were H controls. However, MS and control females did not differ in auditory startle response or in startle-induced ultrasonic vocalizations. Therefore, experiencing maternal separation results in a long-lasting increase in anxiety-like behaviors that occurs in a sex-dependent manner. PMID- 12076733 TI - Effect of a single maternal separation at different pup ages on the corticosterone stress response in adult and aged rats. AB - Postnatal days (PNDs) 4-14 constitute the stress hyporesponsive period (SHRP) of the rat's pituitary-adrenal axis. The impact of manipulation of the pup-dam relationship during the SHRP on neuroendocrine and behavioural function has been the subject of considerable investigation. A single period of 24-h separation of the litter from the dam (maternal separation, MS) during the SHRP increases pup pituitary-adrenal activity and attenuates the SHRP. The MS manipulation also allows for the age-specific analysis of the chronic effects of early-life stress. Here we report on the effects of MS performed at the beginning of (PND 4), or about midway into (PND 9), or after (PND 18) the SHRP, on basal and stress related blood corticosterone (CORT) titers in mature (month 5) and old (month 20) adult males. MS at PND 4, 9, or 18 did not affect basal CORT plasma titers. MS at each of these ontogenetic stages led to a similar and significant increase in the CORT response to restraint in adults but not in old adults. Therefore, whereas MS exerts a chronic impact on stress-related pituitary-adrenal activity in adult male rats, the effect of this postnatal experience does not depend upon the ontogenetic/SHRP status of the pup, and nor does it persist into senescence. PMID- 12076734 TI - Multiple feedback mechanisms activating corticotropin-releasing hormone system in the brain during stress. AB - Stress-associated disorders such as melancholic depression are characterized by persistent hypothalamic-pituitary-adrenocortical (HPA) axis activation and intensive anxiety. Corticotropin-releasing hormone (CRH) appears to play an essential role in pathophysiology of such disorders. In an attempt to elucidate possible mechanisms underlying persistent activation of CRH in the central nervous system (CNS), we examined responses of hypothalamic and extrahypothalamic CRH systems to the stressors (immobilization stress or psychological stress) and interactions between these CRH systems and glucocorticoids in rats. We propose multiple feedback loops activating central CRH system: (1) attenuation of glucocorticoid-induced negative feedback on the activity of the hypothalamic and brainstem nuclei during chronic stress, (2) autoregulation of CRH biosynthesis in the hypothalamic paraventricular nucleus (PVN) through up-regulation of Type-1 CRH receptor (CRHR-1), and (3) glucocorticoid-mediated positive effects on the amygdaloid CRH system. Stress initially activates the hypothalamic CRH system, resulting in the hypersecretion of glucocorticoids from the adrenal gland. In addition, the psychological component of the stressor stimulates the amygdaloid CRH system. In the chronic phase of stress, down-regulation of GR in the PVN and other brain structures such as the locus coeruleus (LC) fails to restrain hyperfunction of the HPA axis, and persistent activation of the HPA axis further up-regulates the amygdaloid CRH system. Thus, the hypothalamic and the amygdaloid CRH systems cooperatively constitute stress-responsive, anxiety-producing neurocircuitry during chronic stress, which is responsible for the clinical manifestations of stress-associated disorders. Effects of tricyclic antidepressants (TCAs), which appear to mitigate the above mentioned multiple feedback loop forming the vicious circle to activate central CRH systems, will also be discussed. PMID- 12076735 TI - Stress and nerve growth factor: findings in animal models and humans. AB - Stress is elicited by environmental, social or pathological conditions occurring during the life of animals and humans that determine changes in the nervous, endocrine and immune systems. In the present review, we present data supporting the hypothesis that stress-related events both in animal models and humans are characterized by modifications of endogenous nerve growth factor (NGF) synthesis and/or utilization. Stress inducing alteration in NGF synthesis and/or utilization appears to be more severe during neurogenesis and in early postnatal life. However, NGF endogenously released during stress may promote remodeling of damaged tissues following acute and/or chronic stressful events. PMID- 12076736 TI - Environmental influences on brain neurotrophins in rats. AB - Environmental factors can have profound influences on the brain. Enriching environments with physical, social and sensory stimuli are now established to be beneficial to brain development and ageing. A multitude of responses from cellular and molecular mechanisms to macroscopic changes in neural morphology and neurogenesis have been considered in the context for evidences that environmental inputs can regulate brain plasticity in the rat at all stages of life. Data from our laboratory have revealed that enriched environment increased nerve growth factor (NGF) gene expression and protein levels in the hippocampus, and this may contribute to events underlying environmentally induced neural plasticity. Because neurotrophic factors are essential for neural development and survival, they are likely to be involved in the cerebral consequences modified by enriched experiences. PMID- 12076737 TI - Early maternal deprivation and prepulse inhibition: the role of the postdeprivation environment. AB - Early postnatal maternal deprivation leads to a variety of biochemical and behavioural alterations in the offspring, some of which do not develop until adulthood, like deficits in prepulse inhibition. Since a number of these deficits are similar to abnormalities observed in schizophrenic patients, maternal deprivation has been proposed as an interesting model for schizophrenia. However, little is still known about the processes that determine these long-term consequences. Previous experiments showed that the strain of rats and the deprivation procedure are important factors. In the present set of experiments, we focussed on the postdeprivation period. We showed that rearing normal Wistar rats in social isolation from weaning disrupts prepulse inhibition. However, if maternally deprived Wistar rats were reared in social isolation, the prepulse inhibition was normal. We further showed that if only half of the litters were maternally deprived at postnatal day 9, the animals had only a small disruption in prepulse inhibition compared to animals that came from litters where all the animals were deprived. In a final experiment, we crossfostered maternally deprived mothers to nondeprived pups and vice versa. This experiment showed that both the nondeprived pups raised by a deprived mother and the deprived pups raised by a nondeprived mother had small deficits in prepulse inhibition. Taken together, these data clearly show that the postdeprivation period is of crucial importance for the development of prepulse inhibition deficits in maternally deprived rats. We present a working model in order to explain the long-term behavioural consequences of maternal deprivation. PMID- 12076738 TI - The effects of isolation rearing on glutamate receptor NMDAR1A mRNA expression determined by in situ hybridization in Fawn hooded and Wistar rats. AB - Rats reared in social isolation exhibit a syndrome of behavioral and biochemical effects indicative of enhanced mesolimbic dopamine (DA) function. The precise nature of the neurodevelopmental changes that produce this state are unknown but result in enhanced DA neurotransmission in the nucleus accumbens (NAC). It was hypothesized that this may be the indirect result of chronic changes in glutamate NMDA receptor function. The same prediction has been made for Fawn hooded (FH) rats that exhibit some of the characteristic effects of isolation-reared rats when compared to Wistar rats. Therefore, mRNA levels of the NMDAR1A receptor subunit were determined by in situ hybridization and were quantified in the striatum, hippocampus and prefrontal cortex of FH and Wistar rats. Isolation rearing alone was not found to have an effect on the expression of NMDAR1A, while FH rats had reduced levels across most brain regions examined. In some areas of the striatum and prefrontal cortex, this effect was greater in FH isolates than in FH socials, while in the hippocampus, the opposite was observed. PMID- 12076739 TI - Psychological stress and environmental adaptation in enriched vs. impoverished housed rats. AB - In this study, we report differential behavioural and cognitive effects, as assessed in the open-field and the Morris water maze, following psychological stress in enriched vs. impoverished housed rats. Three stress conditions were evaluated: nonstress, mild stress and powerful stress. Mild stress consisted of exposure to an avoidance box but without shock, while in the powerful stress condition animals were exposed to an electric shock. The results revealed distinct effects in the differentially housed animals. Prior exposure to a mild stress enhanced escape performance in the water maze in enriched but not impoverished animals. However, preexposure to powerful stress negatively affected animals from both housing conditions in the water maze task, but with the enriched animals less affected than impoverished animals. In the open-field test, stress preexposure reduced locomotion counts in both the differentially housed animals. In addition, the results showed that the enrichment effect on emotional reactivity in the open-field is long-lasting and persists even after extensive training and housing in standard laboratory conditions. The results are discussed in relation to the nature of the behavioural and learning differences between the differentially housed animals. PMID- 12076740 TI - Dissociable effects of isolation rearing and environmental enrichment on exploration, spatial learning and HPA activity in adult rats. AB - Male Lister hooded rats were reared from weaning either singly or in groups of three in either barren or enriched cages (n=9 each) to study effects of isolation rearing and environmental enrichment on open-field activity, object exploration, activity in the Light/Dark box (L/D box), spatial learning and memory in the Morris water maze, and hypothalamic-pituitary-adrenal (HPA) activity in response to restraint stress. Regardless of inanimate background, isolation rearing mainly enhanced activity under several conditions of environmental novelty. By contrast, environmental enrichment, regardless of social background, primarily accelerated habituation to novelty and improved spatial learning and memory. None of the treatments significantly altered basal and response levels of plasma ACTH and corticosterone. Furthermore, rats reared singly in barren cages showed persistent activity in the L/D box, indicating an interaction between isolation-induced hyperactivity and reduced habituation due to barren caging. These results show that isolation rearing and environmental enrichment affect behaviour selectively, while at the same time revealing biologically relevant interactions between social and inanimate stimulation. It is concluded that systematic variation of social and inanimate stimulation can help distinguish between effects that generalise across variation in environmental background and effects that are idiosyncratic to a specific environmental background. PMID- 12076741 TI - Enduring effects of environmental enrichment on novelty seeking, saccharin and ethanol intake in two rat lines (RHA/Verh and RLA/Verh) differing in incentive seeking behavior. AB - The Roman high- and low-avoidance (RHA/Verh and RLA/Verh) rat lines represent, respectively, low emotional/anxious and high novelty seeker vs. high emotional/anxious and low novelty seeker profiles. In the present study, RLA/Verh and RHA/Verh rats, either reared in pairs from weaning (untreated) or reared in groups of 8-10 in an enriched environment until the age of 7 months, were tested for exploratory and novelty-seeking behavior in the hole board (including novel objects under the holes), as well as for their preference for saccharin-water and ethanol-water in a two-bottle free-choice paradigm. Testing started when rats were 20 months old in order to study the long-lasting effects of differential rearing. RHA/Verh rats explored more and showed greater preference for (and intake of) saccharin as well as for ethanol than RLA/Verh rats, thus confirming their validity as a rat model for sensation/reward seeking. Environmental enrichment (EE) increased head-dipping behavior (i.e., novelty seeking) in both rat lines, without affecting locomotor activity. EE treatment increased the preference for, and volume intake of, saccharin (especially at the higher concentrations tested) in the relatively low saccharin-preferring RLA/Verh rats, and also enhanced ethanol consumption in both rat lines. Thus, the results demonstrate consistent and enduring effects of EE on incentive-seeking behavior and further the analysis of how individual differential predispositions for the need of novelty and contact with (or consumption of) rewarding substances arise through either biological (genetic) or early environmental factors, or both. PMID- 12076742 TI - Early-life handling stimulation and environmental enrichment: are some of their effects mediated by similar neural mechanisms? AB - Neonatal (early) handling (EH) and environmental enrichment (EE) of laboratory rodents have been the two most commonly used methods of providing supplementary environmental stimulation in order to study behavioral and neurobiological plasticity. A large body of research has been generated since the 1950s, unequivocally showing that both treatments induce profound and long-lasting behavioral and neural consequences while also inducing plastic brain effects and being "protective" against some age-related deficits. The present work is aimed at reviewing the main neurobehavioral effects of both manipulations, with the final purpose of comparing them and trying to find out to what extent the effects of both treatments may share (or not) possible neural mechanisms. PMID- 12076743 TI - Social stress in tree shrews: effects on physiology, brain function, and behavior of subordinate individuals. AB - Social stress is known to be involved in the etiology of central nervous disorders such as depression. In recent years, animal models have been developed that use chronic stress to induce neuroendocrine and central nervous changes that might be similar to those occurring in the course of the development of depressive disorders. The present review gives a summary of observations made in the tree shrew chronic social stress model. During periods of daily social stress, male tree shrews develop symptoms that are known from many depressed patients such as persistent hyperactivities of both the hypothalamic-pituitary adrenal (HPA) axis and sympathetic nervous system, disturbances in sleeping patterns, and reduced motor activity. Moreover, various physiological parameters indicate an acceleration of the over all metabolic rate in socially stressed tree shrews. Some of these parameters can be renormalized by antidepressants thus supporting the view of the tree shrew social stress paradigm as model for major depression. In the brains of socially stressed animals, monoamine receptors show dynamic changes that reflect adaptation to the persistent monoaminergic hyperactivity during periods of chronic stress. In addition to the changes in neurotransmitter systems, there are structural changes in neurons, e.g., retraction of the dendrites of hippocampal pyramidal neurons. Together, these processes are suggested as a cause of behavioral alterations that can be counteracted by antidepressants in this naturalistic social stress model. PMID- 12076744 TI - Early deprivation and behavioral and physiological responses to social separation/novelty in the marmoset. AB - Long-term effects of adverse early environment on neurobehavioral development have been reported for rodents and primates. The present study used daily early deprivation (ED), a paradigm developed for rats, for the first time in a nonhuman primate, the common marmoset, and investigated its effects on the behavioral and physiological responses to social separation/novelty (SSN) challenge tests in juveniles. On postnatal days (PNDs) 2-28, infants (n=5 twin pairs) were removed from the parents and placed alone in an isolation chamber for 30-120 min (9 h/week). Parents and control subjects (n=5 twin pairs) were briefly restrained (CON). At Weeks 18-20, behavioral responses of ED and CON juveniles to six 60-min SSN tests in an isolated cage, comprising 45 min alone and 15 min reunion with the father, were measured. Baseline and post-test urine samples were collected for measurement of cortisol. ED subjects exhibited significantly lower basal SSN urinary cortisol than CON, whilst SSN response cortisol values were similar in ED and CON. When alone, ED subjects were significantly less mobile and emitted significantly less contact calls than CON. Following reunion, ED subjects were significantly less in contact with or being carried by the father than CON and demonstrated significantly more tail piloerection. Although they require validation by additional parameters (e.g. cardiovascular), these data strongly suggest that early-life stress alters endocrine and behavioral responsiveness to psychosocial challenge in this primate and in a direction that could model important changes in disorders of human affective state. PMID- 12076745 TI - Effects of social manipulations and environmental enrichment on behavior and cell mediated immune responses in rhesus macaques. AB - This paper reviews a series of studies that have examined the effects of manipulations to the social and the inanimate environments on the behavior and cell-mediated immune responses of rhesus macaques of various ages living in different settings. In general, enrichment of the inanimate environment with toys, structures, foraging devices, and/or videotapes increased the amount of species-typical behavior expressed by the monkeys, but did not affect their immune responses. Housing monkeys socially, on the other hand, not only resulted in increased time spent in species-typical activities, but also resulted in (1) decreases in time spent in abnormal behavior and (2) changes in a number of immune parameters. Additionally, attempts to directly influence the affiliative interactions of socially housed adult rhesus have resulted in systematic changes in affiliative behavior, although anticipated accompanying systematic alterations to cell-mediated immune responses have yet to be realized. The data suggest that aspects of the physical and social environments influence behavioral and immunological parameters in captive macaques in the absence of other experimental manipulations. As such, these influences need to be appropriately managed and/or controlled in order to minimize potential confounds in experimental designs. PMID- 12076746 TI - Environmental enrichment for the captive spectacled bear (Tremarctos ornatus). AB - As part a series of investigations of environmental enrichment methods for zoo animals, two spectacled bears (Tremarctos ornatus) were observed for 40 h, documenting use of cage space and behaviors, using a detailed ethogram. Baseline data showed concentration of activity into limited areas of the enclosure and expression of a relatively restricted subset of the species' behavioral repertoire. Introduction of a climbing structure resulted in increased behavioral diversity, both in the use of the enclosure's physical space and the behaviors displayed in various parts of the enclosure. PMID- 12076747 TI - Cidofovir in the treatment of poxvirus infections. AB - Cidofovir [(S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine, HPMPC] has since 1996 been licensed for clinical use in the treatment of cytomegalovirus (CMV) retinitis in AIDS patients. Cidofovir has broad-spectrum activity against virtually all DNA viruses, including herpes-, adeno-, polyoma-, papilloma- and poxviruses. Among the poxviruses, vaccinia, variola (smallpox), cowpox, monkeypox, camelpox, molluscum contagiosum and orf have proven sensitive to the inhibitory effects of cidofovir. In vivo, cidofovir has shown high efficacy, even after administration of a single systemic (intraperitoneal) or intranasal (aerosolized) dose, in protecting mice from a lethal respiratory infection with either vaccinia or cowpox. Cidofovir has also demonstrated high effectiveness in the treatment of vaccinia virus infection in severe combined immune deficiency mice. In humans, cidofovir has been used successfully in the treatment, by both the topical and intravenous route, of recalcitrant molluscum contagiosum and orf in immunocompromised patients. Taken together, these data indicate that cidofovir should be effective in the therapy and short-term prophylaxis of smallpox and related poxvirus infections in humans, as well as the treatment of the complications of vaccinia that may arise in immunocompromised patients inadvertently inoculated with the smallpox vaccine (vaccinia). PMID- 12076748 TI - Goals and milestones during treatment of HIV-1 infection with antiretroviral therapy: a pathogenesis-based perspective. AB - Highly active antiretroviral therapy (HAART) has reduced the morbidity and mortality related to infection with the human immunodeficiency virus-1 (HIV-1) through its ability to suppress viral replication and preserve and reconstitute specific immune responses in many infected individuals. However, the complete eradication of HIV-1 with current HAART regimens is not considered possible by most experts. Moreover, many current antivirals have metabolic complications and limiting side effects. Consequently, the treatment paradigm has shifted from 'hit hard and early' to delaying the initiation of therapy until later in the course of HIV-1-related disease, with corresponding modifications of consensus treatment guidelines. Factors that need to be considered in deciding when to initiate therapy and with what regimen include the patient's risk of disease progression, the possible adverse drug effects, the patient's ability to adhere to the prescribed therapy, and the need to preserve future therapeutic options. In this article, we discuss the issues surrounding the initiation of HAART, and describe the virologic and immunologic milestones that may be achieved with effective antiretroviral therapy. PMID- 12076749 TI - Characterizing antiviral activity of adefovir dipivoxil in transgenic mice expressing hepatitis B virus. AB - Oral adefovir dipivoxil (ADV) reduced viral load in transgenic mice expressing hepatitis B virus (HBV). Liver HBV DNA was reduced to <0.1 pg of viral DNA per microg of total DNA (pg/microg) following oral ADV therapy at a dosage of 100 mg/kg/day twice daily for 10 days as compared to a mean of 3.0 pg/microg for the placebo control group. Oral ADV treatment also reduced serum HBV DNA to 3.5 log10 genomic equivalents (ge)/ml compared to 5.3 log10 ge/ml for the placebo control group. With once daily treatments, ADV antiviral activity reached near maximum viral reduction by day 10 in the liver and reached an endpoint of liver virus inhibition at 1.0 mg/kg/day. The minimum effective dose was less than 0.1 mg/kg/day using inhibition of serum virus. Lamivudine (3TC) given orally at 500 mg/kg/day using the same treatment schedule marginally reduced the serum HBV DNA by 4-fold, but did not significantly reduce HBV liver DNA. Serum titer reduction was also identified in untreated or placebo-treated animals, which may have been caused by the stress of pre-treatment bleeding and multiple oral gavage treatments. This trauma/placebo-effect may have masked the extent of viral reduction in the serum in ADV- and 3TC-treated animals. Liver HBV RNA was not reduced by oral ADV treatments. The lack of RNA reduction was expected, because the HBV transgene is stably integrated into the chromosome and ADV inhibits polymerase activity after transcription of pregenomic RNA. ADV was identified to have potent anti-HBV activity in this HBV transgenic mouse model and could serve as a suitable positive control for future drug discovery experiments. PMID- 12076750 TI - Dose proportional inhibition of HIV-1 replication by mycophenolic acid and synergistic inhibition in combination with abacavir, didanosine, and tenofovir. AB - Mycophenolate mofetil (MMF), a therapeutically used inhibitor of inosine monophosphate dehydrogenase is hydrolyzed to its active metabolite mycophenolic acid (MPA) in vivo. MPA exhibits anti-HIV activity in vitro. We tested MPA alone and in combination with abacavir (ABC), didanosine (DDI), lamivudine (3TC) and tenofovir (TFV) against wild-type human immunodeficiency virus type-1 (HIV-1) and nucleoside reverse transcriptase inhibitor (NRTI)-resistant HIV-1. MPA (62.5-500 nM), when combined with ABC or DDI, synergistically enhanced activity against wild-type HIV and the NRTI-resistant HIV clone DRSM34. MPA also enhanced the activity of TFV against both wild-type HXB2 and TFV-resistant strain HIV(K65R), in a more than additive manner. No significant antiproliferative effect of MPA (< or =0.25 microM) alone or in the presence of ABC, DDI and TFV was observed. This indicates that the antiviral effects of MMF may be clinically achievable without fully blocking T-cell proliferation or inducing immunosuppression. These findings provide further rationale for the clinical testing of MMF in combination with ABC, DDI, and TFV. PMID- 12076752 TI - Metabolic pathways of N-methanocarbathymidine, a novel antiviral agent, in native and herpes simplex virus type 1 infected Vero cells. AB - N-methanocarbathymidine ((N)-MCT), a thymidine analog incorporating a pseudosugar with a fixed Northern conformation, exhibits potent antiherpetic activity against herpes simplex virus types 1 (HSV-1) and 2 (HSV-2). This study contrasts the metabolic pathway of (N)-MCT and the well-known antiherpetic agent ganciclovir (GCV) in HSV-1-infected and uninfected Vero cells. Treatment of HSV-1 infected Vero cells immediately after viral infection with (N)-MCT profoundly inhibited the development of HSV-1 infection. Using standard plaque reduction assay to measure viral infection, (N)-MCT showed a potency greater than that of ganciclovir (GCV), the IC50s were 0.02 and 0.25 microM for (N)-MCT and GCV, respectively. (N)-MCT showed no cytotoxic effect on uninfected Vero cells (CC50>100 microM). Dose and time dependence studies showed high levels of (N)-MCT triphosphate ((N)-MCT-TP), and GCV-triphosphate (GCV-TP) in HSV-1-infected cells incubated with (N)-MCT or GCV, respectively. In contrast, uninfected cells incubated with (N)-MCT showed elevated levels of (N)-MCT-monophosphate only, while low levels of mono, di- and triphosphates of GCV were found following incubation with GCV. Although the accumulation rate of (N)-MCT and GCV phosphates in HSV-1-infected cells were similar, the decay rate of (N)-MCT-TP was slower than that of GCV-TP. These results suggest that: (1) the antiviral activity of (N)-MCT against herpes viruses is mediated through its triphosphate metabolite; (2) in contrast to GCV, the diphosphorylation of (N)-MCT in HSV-1- infected cells is the rate limiting step; (3) (N)-MCT-TP accumulates rapidly and has a long half life in HSV-1-infected cells; and (4) HSV-tk catalyzed the mono, and diphosphorylation of (N)-MCT while monophosphorylating GCV only. These results provide a biochemical rational for the highly selective and effective inhibition of HSV-1 by (N)-MCT. PMID- 12076751 TI - Antiviral activity of Plantago major extracts and related compounds in vitro. AB - Plantago major L., a popular traditional Chinese medicine, has long been used for treating various diseases varying from cold to viral hepatitis. The aim of present study was to examine the antiviral activity of aqueous extract and pure compounds of P. major. Studies were conducted on a series of viruses, namely herpesviruses (HSV-1, HSV-2) and adenoviruses (ADV-3, ADV-8, ADV-11). The antiviral activity of EC50 was defined as the concentration achieved 50% cyto protection against virus infection and the selectivity index (SI) was determined by the ratio of CC50 (concentration of 50% cellular cytotoxicity) to EC50. Results showed that aqueous extract of P. major possessed only a slight anti herpes virus activity. In contrast, certain pure compounds belonging to the five different classes of chemicals found in extracts of this plant exhibited potent antiviral activity. Among them, caffeic acid exhibited the strongest activity against HSV-1 (EC50=15.3 microg/ml, SI=671), HSV-2 (EC50=87.3 microg/ml, SI=118) and ADV-3 (EC50=14.2 microg/ml, SI=727), whereas chlorogenic acid possessed the strongest anti-ADV-11 (EC50=13.3 microg/ml, SI=301) activity. The present study concludes that pure compounds of P. major, which possess antiviral activities are mainly derived from the phenolic compounds, especially caffeic acid. Its mode of action against HSV-2 and ADV-3 was found to be at multiplication stages (postinfection of HSV-1: 0-12 h; ADV-3: 0-2 h), and with SI values greater than 400, suggesting the potential use of this compound for treatment of the infection by these two viruses. PMID- 12076753 TI - Additive and antagonist effects of therapeutic gene combinations for suppression of HIV-1 infection. AB - A previously described Moloney-based vector expressing a double copy anti-tat antisense tRNA (DC-tRNA-AT) (Biasolo et al., 1996. J. Virol. 70, 2154-2161) was modified to increase the copy number of the antisense molecule and to target the intra-cytoplasmic localization of the HIV genome. To this end, an anti-U5 hammerhead ribozyme, engineered as a hybrid small adenoviral VAI RNA (VAIalpha), was inserted into the vector as a single molecule or in combination with the double copy anti-tat sequence. The retroviral vector expressing only VAIalpha (DC VAIalpha) inhibited HIV-1 replication to an extent comparable to that of DC-tRNA AT. A more effective inhibition was produced by the vector expressing multiple copies of the anti-tat antisense (DC-6tRNA-AT). This higher effectiveness correlated with anti-tat stochiometry, i.e. with the absolute number of therapeutic molecules being produced on a per cell basis at the steady state. Surprisingly, when the tRNA-AT and VAIalpha genes were combined in the same vector (DC-AT-VAIalpha), an enhancement of viral replication was noticed. This study indicates that it is possible to potentiate the antiviral activity of a retroviral vector by increasing the steady-state level of the therapeutic molecule. Results also show that the combined expression of two singularly active therapeutic RNAs can have antagonistic rather than synergistic effects. PMID- 12076754 TI - Isolation of two highly potent and non-toxic inhibitors of human immunodeficiency virus type 1 (HIV-1) integrase from Salvia miltiorrhiza. AB - Water soluble extracts of the herbal plant, Salvia miltiorrhiza (Danshen) exhibited potent effect against HIV-1 integrase activity in vitro and viral replication in vivo. We have developed an extensive purification scheme to isolate effective, non-toxic inhibitors against human immunodeficiency virus type 1 (HIV-1) using the 3'-processing activity of integrase as a purification guide and assay. Two water soluble compounds, M(5)22 and M(5)32, have been discovered by isolating them from S. miltiorrhiza roots in purities of >99.5% as shown by NMR spectral analysis with yields of 0.018 and 0.038%, respectively. Structural determination revealed that M(5)22 is lithospermic acid and M(5)32 is lithospermic acid B. These two structurally related compounds are potent anti-HIV inhibitors and showed no cytotoxicity to H9 cells at high concentrations (CC(100)>297 microM for M(5)22 and >223 microM for M(5)32). The IC50 for inhibition of 3'-processing by HIV-1 integrase was found to be 0.83 microM for M(5)22 and 0.48 microM for M(5)32. In addition, M(5)22 and M(5)32 inhibited HIV-1 integrase catalytic activities of 3'-joining to the target DNA with IC50 of 0.48 microM for M(5)22 and 0.37 microM for M(5)32. Furthermore, kinetic and mechanistic studies suggested that drug binding to HIV-1 integrase and inhibition of enzymatic activity occur at a fast rate. Both M(5)22 and M(5)32 do not prevent HIV entry in H9 cells. They also show no inhibition of reverse transcriptase activity in infected cells. The levels of intracellular strong stop and full length viral DNA remained unchanged following drug treatment. However, both inhibitors strongly suppressed the acute HIV-1 infection of H9 cells with IC50 values of 2 and 6.9 microM for M(5)22 and M(5)32, respectively. Thus these two selective integrase inhibitors hold promise as a novel class of therapeutic drugs for AIDS based on their high potencies and absence of cytotoxicity. PMID- 12076756 TI - Synthesis, cytotoxicity and antiviral activity of N,N'-bis-5-nitropyrimidyl derivatives of dispirotripiperazine. AB - During the search for new antivirals, various N,N'-bis-5-pyrimidyl derivatives of 3,12-diaza-6,9-diazonia(5,2,5,2)dispirohexadecane dichloride (dispirotripiperazine) were synthesized. To reveal relationships between chemical structure and antiviral activity, the compounds were characterized by fast atom bombardment mass, nuclear magnetic resonance, infra red spectroscopy, and elemental analysis and examined for cytotoxicity, inhibition of cell growth and antiviral activity under in vitro conditions. The results of this study demonstrate an excellent compatibility of the test compounds for confluent as well as proliferating cells and a potent structure-dependent inhibition of herpes simplex virus type 1 replication when added during viral adsorption. Functional group analysis revealed that both the dispirotripiperazine as well as the pyrimidine ring with a nitro group in the 5 position are necessary for activity. A reduction of electron density in the terminal pyrimidine rings enhanced the antiviral activity whereas electron donor substitutions reduced it. Introduction of a methyl group in position 2 of the pyrimidine had no influence on cytotoxicity or antiviral activity. PMID- 12076755 TI - Identification of active antiviral compounds against a New York isolate of West Nile virus. AB - The recent West Nile virus (WNV) outbreak in the United States has increased the need to identify effective therapies for this disease. A chemotherapeutic approach may be a reasonable strategy because the virus infection is typically not chronic and antiviral drugs have been identified to be effective in vitro against other flaviviruses. A panel of 34 substances was tested against infection of a recent New York isolate of WNV in Vero cells and active compounds were also evaluated in MA-104 cells. Some of these compounds were also evaluated in Vero cells against the 1937 Uganda isolate of the WNV. Six compounds were identified to be effective against virus-induced CPE with 50% effective concentrations (EC50) less than 10 microg/ml and with a selectivity index (SI) of greater than 10. Known inhibitors of orotidine monophosphate decarboxylase and inosine monophosphate dehydrogenase involved in the synthesis of GTP, UTP, and TTP were most effective. The compounds 6-azauridine, 6-azauridine triacetate, cyclopententylcytosine (CPE-C), mycophenolic acid and pyrazofurin appeared to have the greatest activities against the New York isolate, followed by 2-thio-6 azauridine. Anti-WNV activity of 6-azauridine was confirmed by virus yield reduction assay when the assay was performed 2 days after initial infection in Vero cells. The neutral red assay mean EC50 of ribavirin was only 106 microg/ml with a mean SI of 9.4 against the New York isolate and only slightly more effective against the Uganda isolate. There were some differences in the drug sensitivities of the New York and Uganda isolates, but when comparisons were made by categorizing drugs according to their modes of action, similarities of activities between the two isolates were identified. PMID- 12076757 TI - Multitarget ribozyme against the S1 genome segment of reovirus possesses novel cleavage activities and is more efficacious than its constituent mono-ribozymes. AB - Two hammerhead motif containing ribozymes (Rzs) were constructed through recombinant techniques that were directed to cleave at the conserved sites of the reovirus S1 gene segment which encodes the cell attachment protein sigma1. The two mono-ribozymes 553 and 984 cleaved the target RNA in a sequence specific manner, and Rz-553 being the more efficient. When the mono-Rzs were combined in direct tandem to make it a multitarget-Rz, very efficient cleavage of the S1 RNA was achieved that retained the specificity of the two mono-ribozymes. This cleavage was, as expected, Mg(++)-dependent but protein-independent. Almost complete cleavage of the S1 RNA was observed with multitarget ribozyme alone. Although S1-Rz-984 cleaved the short S1 synthetic RNA, it failed to cleave the full length S1 RNA (1.4 kb). On the contrary, Rz-553 cleaved the short synthetic RNA as well as the full length S1 RNA with equal efficiency. Full length S1 RNA was, however, cleaved efficiently by the multitarget-ribozyme-S1-Rz-984-553 that cleaved at both the target sites. Thus, hybridization of one ribozyme (Rz-553) to a full length S1 RNA potentially opened up the 984-Rz target site that was otherwise inaccessible to the mono-Rz-984. Multitarget ribozyme expressing mammalian cells showed reduced amounts of S1 RNA that correlated well with the levels of reovirus sigma1 protein. Potential uses of such multitarget-Rzs are discussed. PMID- 12076758 TI - Mutations in the conserved woodchuck hepatitis virus polymerase FLLA and YMDD regions conferring resistance to lamivudine. AB - During more than 104 weeks of treatment with lamivudine (3TC) in chronic woodchuck hepatitis virus (WHV) carrier woodchucks, viral recrudescence occurred. Analysis of WHV DNA polymerase from woodchuck serum samples by PCR followed by DNA sequencing demonstrated that all samples were wild type at the conserved YMDD motif in domain C. Four of the six 3TC-treated woodchucks showed a mixture of the wild-type Ala (GCT) and the mutant Thr (ACT) at the conserved amino acid residue 566 (FLLA) in domain B of the WHV polymerase region. The appearance of the A566T mutation was temporally associated with viral recrudescence. This change is analogous with the amino acid 181 (FLLA) in HBV where 3TC selects for a change from Ala to Thr in humans. In the woodchuck, the Ala to Thr change in the polymerase gene results in a mutation of the WHV surface protein (amino acid 377) from Trp (TGG) to an opal codon (TGA), which may prematurely terminates the polypeptide. Three WHV molecular infectious clones were constructed to study this mutation in greater detail in vitro: A566T, analogous to A181T in HBV; M589V, analogous to the M204V in HBV; and the double mutant A566T/M589V, analogous to A181T/M204V in HBV. These mutants exhibited drug-sensitivity and replication profiles that paralleled those reported for analogous HBV variants. In transfected Huh7 cells, WHV containing the M589V mutation conferred at least 100 fold increased resistance to 3TC, but replicated approximately 5-fold less efficiently than wild-type virus as judged by both extracellular virus production and intracellular DNA replicative forms. In contrast, A566T mutant was approximately 10-fold more resistant to 3TC, replicated intracellularly as well as wild type, but produced 10-fold lower levels of virions than wild type. These findings are consistent with the observation that the A566T mutation alters the overlapping WHV surface antigen reading frame. WHV carrying mutations in the conserved YMDD motif, while not directly selected during lamivudine therapy in WHV carrier woodchucks, are replication competent in cell culture indicating the potential for their emergence in treated animals. These results further illustrate the utility of the WHV/woodchuck model to studies of HBV-drug resistance. PMID- 12076759 TI - 3-deazaneplanocin A induces massively increased interferon-alpha production in Ebola virus-infected mice. AB - 3-deazaneplanocin A, an analog of adenosine, is a potent inhibitor of Ebola virus replication. A single dose early in infection prevents illness and death in Ebola virus-infected mice. The ability of this and similar compounds to block both RNA and DNA viruses has been attributed to the inhibition of a cellular enzyme, S adenosylhomocysteine hydrolase (SAH), indirectly resulting in reduced methylation of the 5' cap of viral messenger RNA. However, we found that the protective effect of the drug resulted from massively increased production of interferon alpha in Ebola-infected, but not uninfected mice. Peak interferon levels increased with the extent of disease at the time of treatment, indicating that production was boosted only in virus-infected cells. Ebola virus has been shown to suppress innate antiviral mechanisms of the type I interferon response. 3 deazaneplanocin A appears to reverse such suppression, restricting viral dissemination. Further development should focus on identifying adenosine analogues that produce a similar effect in Ebola virus-infected primates. PMID- 12076760 TI - A mammalian two-hybrid screening system for inhibitors of interaction between HIV Nef and the cellular tyrosine kinase Hck. AB - In the scope of the search for new anti-HIV agents interacting with a new target, we developed a high-throughput screening system to detect the interactions between Nef and Hck. Nef is an accessory protein of HIV, which is involved in the pathogenicity of the acquired immunodeficiency syndrome (AIDS). Nef is also a signaling molecule because it binds to many host molecules. It has especially high affinity to Hck, a member of src family tyrosine kinase. Using a mammalian two-hybrid system, the interaction between Nef and the SH3 domain of Hck induced luciferase activity with high sensitivity and a Nef-PXXP peptide inhibited this interaction; and so did the anticancer drug adriamycin. We also developed another assay system by coexpression of full-length Hck and Nef, and found that Hck kinase was activated depending on the dose of Nef plasmid. Using the second system, we found that adriamycin interfered with the Nef-Hck interaction by reducing the amount of the Hck protein. The mammalian two-hybrid system may show utility in screening inhibitors of Nef-Hck interaction. PMID- 12076761 TI - Adenovirus inhibition by peptidomimetic integrin antagonists. AB - Many viruses and bacterial pathogens are capable of exploiting host cell surface integrins during their replication cycles. The ligands for many integrins contain an arginine-glycine-aspartic acid (RGD) amino acid sequence that is essential for protein-protein interaction. Human adenovirus particles contain this sequence in the penton base protein, and previous studies support a role for this RGD in integrin-dependent internalization of the virus by the cell. As synthetic peptidomimetics of RGD have been shown in other experimental systems to be antagonists of the activities of specific integrins both in vitro and in vivo, we sought to determine whether these small molecules are antagonists of adenovirus infection. Such compounds inhibited viral infection of cultured cells with similar rank order potency to that determined in assays utilizing purified extracellular matrix proteins as integrin ligands. The maximal level of inhibition achieved with the peptidomimetics was comparable to that of RGD containing peptides, whereas no significant effects were apparent with an RGE containing peptide. An engineered adenovirus having a mutated RGD sequence in the penton base was not susceptible to the inhibition. The results obtained with these synthetic antagonists, which have varied structures and potencies, suggest that integrins interact with adenoviral RGD in a manner similar to that of other protein ligands such as vitronectin. Furthermore, the results confirm the role of RGD in the replication cycle, and suggest peptidomimetic compounds may be useful antimicrobial agents in the treatment of a variety of diseases. PMID- 12076762 TI - Antiviral and immunomodulatory activity of the metal chelator ethylenediaminedisuccinic acid against cytomegalovirus in vitro and in vivo. AB - Antiviral activity of the metal chelator ethylenediaminedisuccinic acid (EDDS) was examined in vitro against human cytomegalovirus (HCMV) wild type strains and strains that are resistant against ganciclovir (GCV) and cidofovir (HPMPC). EDDS inhibited the replication of wild-type as well as GCV- and HPMPC-resistant strains with a 50% effective concentration of 7.4-12 microg/ml. At concentrations of 100 microg/ml EDDS, unlike GCV or HPMPC, suppressed HCMV-induced up-regulation of intercellular adhesion molecule-1 (ICAM-1) and reduced T-cell adhesion to HCMV infected cells in a monolayer adhesion model. In vitro EDDS inhibited murine cytomegalovirus (MCMV) replication (EC50 8.6 microg/ml) and caused in mice some protection against MCMV induced mortality at a non-toxic dose. Since immunopathological factors may play a significant role in HCMV disease it will be of interest to further study whether EDDS is effective in terms of modulation of inflammatory responses to HCMV infections. PMID- 12076763 TI - RSV entry inhibitors block F-protein mediated fusion with model membranes. AB - RSV fusion is mediated by F-protein, a major viral surface glycoprotein. CL 309623, a specific inhibitor of RSV, interacts tightly with F-protein, which results in a hydrophobic environment at the binding site. The binding is selective for F-protein and does not occur with G-protein, a surface glycoprotein that facilitates the binding of RSV to target cells, or with lipid membranes at concentrations in the sub-millimolar range. Using an assay based on the relief of self-quenching of octadecyl rhodamine (R18) incorporated in the RSV envelope, we show that the virus fuses efficiently with large unilamellar vesicles containing cholesterol, in the absence of specific receptor analogs. Fusion of cp-52, a mutant virus lacking the G and SH surface glycoproteins, with vesicles is inhibited by CL-309623 and RFI-641 due to specific interactions of the inhibitor(s) with the fusion protein. Both virus-vesicle and virus-cell fusion are inhibited with equal potency. The formation of the binary complex of CL 309623 with F-protein in its native state, resulting in the inhibition of fusion and entry of virus, is a prerequisite for the observed anti-RSV activity in cell cultures. PMID- 12076764 TI - Polymeric inhibitor of influenza virus attachment protects mice from experimental influenza infection. AB - Synthetic sialic acid-containing macromolecules inhibit influenza virus attachment to target cells and suppress the virus-mediated hemagglutination and neutralize virus infectivity in cell culture. To test the protective effects of attachment inhibitors in vivo, mice were infected with mouse-adapted influenza virus A/Aichi/2/68 (H3N2) and treated with synthetic polyacrylamide-based sialylglycopolymer PAA-YDS bearing moieties of (Neu5Acalpha2-6Galbeta1 4GlcNAcbeta1-2Manalpha1)2-3,6Manbeta1-4GlcNAcbeta1-4GlcNAc. Single intranasal inoculations with PAA-YDS 30 min before or 10 min after infection increased the survival of mice (P<0.01). Multiple treatments with aerosolized PAA-YDS on days 2 5 post infection also increased survival (P<0.01), alleviated disease symptoms, and decreased lesions in the mouse lungs. These data suggest that synthetic polyvalent inhibitors of virus attachment can be used for prevention and treatment of influenza. PMID- 12076765 TI - Genetic manipulation of Giardia lamblia. AB - Giardia lamblia is a flagellated protozoan that infects several species including humans and is a major agent of waterborne outbreaks of diarrhea. G. lamblia is also important in the study of basic eukaryotic molecular biology and evolution; however, it has been difficult to employ standard genetic methods in the study of Giardia. Over the past 6 years, two transfection systems were developed and used for the genetic manipulation of G. lamblia. Both systems allow transient or stable transfection of Giardia and/or foreign genes. The DNA-based transfection system allows electroporation of circular or linear plasmid DNA into trophozoites. The RNA virus-based transfection system requires electroporation of in vitro transcribed RNA into GLV-infected trophozoites. Because G. lamblia is one of the most rudimentary eukaryotes, its processes of transcription, translation and protein transport, as well as its metabolic and biochemical pathways, are of interest. Study of these areas will continue to be advanced using transfection in combination with cellular and molecular tools. Several groups have combined these technologies with other techniques to study protein transport and the transcriptional and post-transcriptional regulation of Giardia genes, including encystation-specific and variant surface protein genes. In addition, coupling antisense techniques with transfection has permitted functional knockout of Giardia metabolic genes, allowing Giardia metabolic pathways to be studied. In the near future, both transfection systems will be potent tools in our investigations of the perplexing questions in Giardia biology. PMID- 12076766 TI - Positive and negative effects of deletions and mutations within the 5' flanking sequences of Plasmodium falciparum DNA polymerase delta. AB - The DNA polymerase delta gene from Plasmodium falciparum is associated with a 1.76 kb 5' untranslated region and a promoter containing long homopolymeric (dA:dT) tracts and intrinsic DNA curvature. Here, a comprehensive deletion and mutational analysis of the DNA Pol(delta) upstream sequences has been undertaken to define functionally important regions. Removal of promoter-proximal DNA was shown to upregulate luciferase reporter gene expression and the ATG-proximal portion of the 5' untranslated region was required in conjunction with the promoter for reporter activity. In contrast to the ATG-proximal 5' untranslated region, deletion of the central part of the untranslated region had a positive effect on expression. Disruption of a homopolymeric (dA:dT) tract adjacent to the main transcription start site both derepressed gene expression and reduced the intrinsic curvature of DNA fragments containing this sequence. PMID- 12076767 TI - A cathepsin L protease essential for Caenorhabditis elegans embryogenesis is functionally conserved in parasitic nematodes. AB - Proteolytic enzymes are involved in processes important to development and survival of many organisms. Parasite proteases are considered potential targets of parasite control yet, for most, their precise physiological functions are unknown. Validation of potential targets requires analysis of function. We have recently identified a cathepsin L (CPL) cysteine protease, Ce-CPL-1, which is essential for embryonic development of the free-living nematode Caenorhabditis elegans. We now show that CPL genes closely related to Ce-cpl-1 are expressed in the animal parasitic nematodes Haemonchus contortus, Dictyocaulus viviparus, Teladorsagia circumcincta, Ancylostoma caninum and Ascaris suum, as well as in plant parasitic nematodes. The similarities in gene structure and encoded amino acid sequence indicate that the parasite and C. elegans CPLs are homologous enzymes. We demonstrate functional compensation of the loss of C. elegans cpl-1 by transgenic expression of the H. contortus cpl-1 gene, rescuing the embryonic lethality. These genes may therefore be orthologues, sharing the same function in both species. Targeting of this enzyme has potential in inhibiting development and transmission of parasitic nematodes. In addition, the role of CPL is important to our understanding of nematode development. PMID- 12076768 TI - Plasmodium falciparum MAEBL is a unique member of the ebl family. AB - Malaria is one of the deadliest human diseases and efforts to control it have been difficult due to the protozoan parasites' complex biology. Malaria merozoite invasion of erythrocytes is an essential part of blood-stage infections. The invasion process is mediated by numerous parasite molecules, such as EBA-175, a member of the ebl family of erythrocyte binding proteins. We have identified maebl, an ebl paralogue, in Plasmodium falciparum and found it highly conserved with its orthologues in P. yoelii and P. berghei, but distinct from other Plasmodium ebl. Importantly, the putative MAEBL ligand domains are highly conserved and are similar to AMA-1, but not the consensus DBL ligand domains present in all other ebl. In mature merozoites, MAEBL localized with rhoptry proteins (RhopH2, RAP-1), including surface localization with RhopH2, but not microneme proteins (EBA-175, BAEBL). MAEBL appears as proteolytically processed fragments in P. falciparum parasites. The amino cysteine-rich ligand domains were present primarily in culture supernatants, while the carboxyl cysteine-rich domain adjacent to the transmembrane domain was preferentially isolated from Triton X-100 extracted fractions. These data indicate that the primary structure of maebl is highly conserved among Plasmodium species, while its characteristics demonstrate a function unique among the ebl proteins. PMID- 12076769 TI - Amino terminus of Plasmodium falciparum acidic basic repeat antigen interacts with the erythrocyte membrane through band 3 protein. AB - The acidic basic repeat antigen (ABRA) of Plasmodium falciparum is localised in the parasitophorous vacuole, and associates with the merozoite surface at the time of schizont rupture. By virtue of its protease-like activity, it is implicated in the process of merozoite invasion and schizont rupture, and therefore, possibly interacts with erythrocyte membrane proteins to execute its function during these events. In this study, using Escherichia coli expressed recombinant fragments of ABRA, we have demonstrated that ABRA interacts with red blood cells through its N-terminus. Out of the four human erythrocyte proteins tested, namely, band 3, glycophorin A and B and spectrin, ABRA showed dose dependent and saturable binding with the band 3 protein. This binding was lost on chymotrypsin treatment of erythrocytes or their membrane extract. Studies with the deletion constructs of the N-terminus revealed that the binding domain lies in the cysteine-rich N-proximal region of ABRA. In addition to the recombinant fragments, native ABRA derived from the P. falciparum-infected erythrocytes also showed binding to band 3 protein. Sequencing of the cysteine-rich 528 bp region, amplified from fifteen field isolates of P. falciparum, showed that not only the five cysteines of mature ABRA but also the whole sequence is fully conserved, even at the nucleotide level. This sequence conservation of the N-terminus and its role in RBC binding suggests that this region may be crucial for any putative function of ABRA, therefore emphasising its importance as a vaccine/drug target. PMID- 12076770 TI - A role for tertiary structure in the generation of antigenic diversity and molecular association of the Tams1 polypeptide in Theileria annulata. AB - The major merozoite-piroplasm surface antigen (mMPSA) of Theileria annulata, Tams1, is known to be antigenically diverse. The possession of variable N-linked glycosylation sites and removal of monoclonal antibody 5E1 reactivity by mild periodate treatment suggested, previously, that divergent epitopes may be conferred by secondary modification. This study has shown that monoclonal antibody 5E1 and polyspecific antisera raised against the native protein react against divergent amino acid epitopes that are dependent on a molecular conformation that is sensitive to periodate. Therefore, no experimental evidence exists to confirm the sequence prediction that Tams1 undergoes N-linked glycosylation. Data is also presented indicating that the conformation of the antigen results in presentation of divergent regions on the external surface of the molecule, while conserved regions are more likely to be internal and hidden. In addition, non-reducing SDS-PAGE analysis demonstrated that Tams1 can undergo molecular association to form homo-dimers, trimers and multimers. The potential influence of tertiary structure and inter-molecular association on Tams1 diversity and function is discussed. PMID- 12076771 TI - The microsporidian polar tube: evidence for a third polar tube protein (PTP3) in Encephalitozoon cuniculi. AB - The invasion strategy used by microsporidia is primarily related to spore germination. Small differentiated spores of these fungi-related parasites inject their contents into target cells through the lumen of a rapidly extruded polar tube, as a prerequisite to obligate intracellular development. Previous studies in Encephalitozoon species that infect mammals have identified two major antigenic polar tube proteins (PTP1 and PTP2) which are predicted to contribute to the high tensile strength of the polar tube via an assembly process dependent on disulfide linkages. By immunoscreening of a cDNA library, we found that a novel PTP is encoded by a single transcription unit (3990 bp) located on the chromosome XI of E. cuniculi. PTP3 is predicted to be synthesized as a 1256-amino acid precursor with a cleavable signal peptide. The mature protein lacks cysteine residue and its large acidic core is flanked by highly basic N- and C-terminal regions. Immunolocalization data indicated that PTP3 is involved in the sporoblast-to-spore polar tube biogenesis. A transcriptional up-regulation during sporogony is supported by a strong increase in the relative amount of Ecptp mRNAs within host cells sampled at late post-infection times. To begin to explore polar tube-associated protein interactions, spore proteins were extracted in the presence of SDS and dithiothreitol then incubated with a chemical cross-linker (DSP or sulfo-EGS). A large multimeric complex was formed and shown to contain PTP1, PTP2 and PTP3 with a few other proteins. PTP3 is hypothesized to play a role in the control of the polar tube extrusion as part of a specific response to ionic stimuli. PMID- 12076773 TI - Suppression of secreted acetylcholinesterase expression in Nippostrongylus brasiliensis by RNA interference. PMID- 12076772 TI - Two families of RNA binding proteins from Trypanosoma brucei associate in a direct protein-protein interaction. AB - We have previously reported the identification of two closely related RNA binding proteins from Trypanosoma brucei, termed p34 and p37. The predicted primary structures of the two proteins are highly homologous with one major difference, an 18 amino acid insertion in the N-terminal region of p37. These two proteins are localized to the nucleus based on immunofluorescence microscopy. Recently, we have shown that p34 and p37 interact with T. brucei 5S rRNA. In order to gain further insight into their function, we have utilized protein affinity chromatography and immune capture approaches to identify T. brucei proteins which associate with p34 and p37. We demonstrate here an interaction of both p34 and p37 with the NOPP44/46 proteins, identified in T. brucei as a family of tyrosine phosphorylated RNA binding proteins primarily localized to the nucleolus. This interaction was mapped to the RNA-binding region of p34/p37 and an acidic region of NOPP44/46 by protein affinity chromatography using recombinant deletion constructs of p34 and p37 and yeast two-hybrid analysis. These data may suggest a role for p34 and p37 and NOPP44/46 in the import and/or assembly pathway of T. brucei 5S rRNA in ribosome biogenesis. PMID- 12076774 TI - Dephosphorylation of cyst wall proteins by a secreted lysosomal acid phosphatase is essential for excystation of Giardia lamblia. PMID- 12076775 TI - Reducing requirements for hemoglobin hydrolysis by Plasmodium falciparum cysteine proteases. PMID- 12076776 TI - Characterization of a spliced leader gene and of trans-spliced mRNAs from Taenia solium. PMID- 12076777 TI - A sub-family of common and highly conserved Plasmodium falciparum var genes. PMID- 12076779 TI - Organization of mitochondrial DNA in the basidiomycetous Dioszegia hungarica (Cryptococcus hungaricus) species. AB - The organization of mitochondrial DNA was investigated in the six collection strains of the basidiomycetous yeast Dioszegia hungarica (Cryptococcus hungaricus) isolated so far. Physical and partial functional maps were constructed. Two strains (CBS 6324 and 6576) were identical while three others (CBS 4214, 5124, 6953) differed not only in the distribution of restriction sites but in gene order as well. Results confirm the hypothesis that these five strains are representatives of different closely related species. The sixth strain CBS 6569 revealed a unique mitochondrial genome organization. Its mtDNA separated into eight bands on agarose gel without enzymatic digestion. These molecules carried mitochondrial genes, and RFLP analysis of the four largest molecules using frequently-cutting restriction enzymes (KpnI and SmaI) showed them to have strongly homologous sequences. This unique mtDNA organization was also observed in a strain of Cystofilobasidium capitatum, providing evidence that CBS 6569 belongs to the Cystofilobasidium clade. PMID- 12076780 TI - Cloning of a Ca(2+)/calmodulin-dependent protein kinase gene from the filamentous fungus Arthrobotrys dactyloides. AB - A Ca(2+)/calmodulin-dependent protein kinase (CaMK) gene was cloned and characterized from Arthrobotrys dactyloides, a nematode-trapping fungus. The resulting 373-amino-acid protein, FCaMK, has significant homology to mammalian CaMKs. FCaMK contains a serine/threonine kinase domain followed by a calmodulin binding domain. The activation loop in FCaMK (amino acids 184-199) contains a phosphorylation site at threonine-188, which could be the target of a kinase activator. Truncated FCaMK mutants revealed that amino acids 296-324 are essential for calmodulin binding. An oligopeptide designed from residues 297-324 formed a stable peptide-calmodulin complex of 1:1 stoichiometry. Southern blot analysis detected a single copy of the fcamk gene, suggesting that FCaMK plays an important role in Ca(2+)/calmodulin signaling in A. dactyloides. PMID- 12076781 TI - Characterization of the CaNAG3, CaNAG4, and CaNAG6 genes of the pathogenic fungus Candida albicans: possible involvement of these genes in the susceptibilities of cytotoxic agents. AB - CaNAG3, CaNAG4, and CaNAG6 form a gene cluster with CaNAG1 CaNAG2 and CaNAG5 that are responsible for glucosamine-6-phosphate deaminase, N-acetylglucosamine phosphate deacetylase and N-acetylglucosamine kinase, but their functions largely remain unclear. In this study, Candida albicans cells carrying null mutations in either CaNAG3, CaNAG4, or CaNAG6 were generated and characterized. They showed increased susceptibility to cycloheximide and attenuated virulence in mice. More profound effects were observed when both CaNAG3 and CaNAG4, which are highly related to each other, were disrupted. Thus, it seems that CaNAG3, CaNAG4, and CaNAG6 are involved in drug sensitivity and virulence in C. albicans. PMID- 12076782 TI - Two immunologically distinct types of protofilaments can be identified in Natrialba magadii flagella. AB - We examine distribution of flagellins along multicomponent flagellar filaments (FF) and protofilaments (PF) of the haloalkaliphilic archaeon Natrialba magadii using immunogold electron microscopy. A high specific polyclonal antibody raised to one of the flagellin types bound homogeneously to the undissociated FF along the full length. At the same time both uniformly labelled and completely unlabelled PF, outwardly indistinguishable one from another, were observed. PMID- 12076783 TI - Identification of Weissella species by the genus-specific amplified ribosomal DNA restriction analysis. AB - The Weissella species have been increasingly isolated from many food and meat samples. For rapid and systematic identification of Weissella species, an amplified ribosomal DNA restriction analysis (ARDRA) has been developed. This method employs both polymerase chain reaction (PCR) and restriction pattern analysis: first, PCR using the genus-specific primers that were designed from 16S rDNA sequence produces a 725-bp band only from Weissella species and next, restriction analysis using MnlI, MseI, and BceAI restriction enzymes identifies these Weissella-specific PCR fragments to the species level. This ARDRA method clearly identified six Weissella strains isolated from kimchi as well as 10 validly described Weissella type strains. Therefore, these results suggest that this ARDRA is a rapid and reliable method for identification of Weissella species. PMID- 12076784 TI - A novel method for rapid production and purification of exfoliative toxin A of Staphylococcus aureus. AB - The exfoliative toxins of Staphylococcus aureus are the causative agents of the scalded-skin syndrome. Previously described methods of toxin production and purification require large quantities of culture medium, take a long time and often produce low yields of toxin. A novel method of toxin production and purification using a dialysis sac to separate the culture medium from the staphylococci is described. This method produces up to 12 mg of crude toxin per ml of bacterial cell culture bathing the surface of the dialysis sac within 36 h and almost 10 mg of purified toxin per ml of cell culture within 3 days, in contrast to previous procedures that took over a week to produce 0.1-1.0 mg ml( 1) crude toxin and less than 0.01 mg ml(-1) purified toxin. This rapid method of toxin production should speed up future research into the pathogenesis of the staphylococcal scalded-skin syndrome. PMID- 12076785 TI - Effect of olive oil mill wastewater on extracellular ligninolytic enzymes produced by Phanerochaete flavido-alba. AB - Our previous results have demonstrated that Phanerochaete flavido-alba decoloration, dephenolization and detoxification of olive oil mill wastewater (OMW) were associated with changes in the ligninolytic major exoenzymes accumulated in the cultures. This paper describes the effect of the two main OMW components (monomeric aromatic compounds and a major brownish polymeric pigment), on extracellular P. flavido-alba ligninolytic enzymes. Laccase was the sole ligninolytic enzyme detected in cultures containing monomeric aromatic compounds. Laccase and an acidic manganese-dependent peroxidase (MnPA, pI<2.8) were accumulated in cultures with OMW or polymeric pigment. Also, modified manganese dependent peroxidases were observed mainly in OMW-supplemented cultures. Laccase was more stable to the effect of OMW toxic components and was accumulated in monomeric aromatic-supplemented cultures, suggesting a more important role than manganese-dependent peroxidases in OMW detoxification. Alternatively, MnPA accumulated in cultures containing the polymeric pigment seems to be more essential than laccase for degradation of this recalcitrant macromolecule by P. flavido-alba. PMID- 12076786 TI - Vector engineering to improve a staphylococcal surface display system. AB - A previously developed expression system for surface display of heterologous proteins on the surface of Staphylococcus carnosus employs the secretion signals from a Staphylococcus hyicus lipase and the cell wall anchoring part of Staphylococcus aureus protein A (SpA) to achieve surface display of expressed recombinant proteins. The system has been successfully used in various applications but the vector has not been considered genetically stable enough to allow protein library display applications, which would be of obvious interest. A new set of vectors, differing in size and devoid of a phage f1 origin of replication, were constructed and evaluated in terms of bacterial growth characteristics and vector stability. Furthermore, surface expression of a model surface protein was monitored by an enzymatic whole-cell assay and flow cytometry. The engineered expression vectors demonstrated dramatically improved stability and growth properties and two of the novel vectors demonstrated retained high surface density of the displayed model protein. The flow cytometry was found to be a powerful tool for observing the surface density of displayed heterologous proteins, and would thus be a rational strategy for monitoring the optimisation of any surface display system. The implications of these improved display vectors for future protein library applications are discussed. PMID- 12076787 TI - Role of fibronectin in curli-mediated internalization. AB - Curli fibers of Escherichia coli mediate internalization of bacteria by eukaryotic cells. As curli fibers bind fibronectin with high affinity, the role of fibronectin in the uptake process was studied. The experiments presented here support the involvement of fibronectin in internalization of bacteria. Furthermore, a peptide containing the RGD motif, responsible for interaction of fibronectin with cellular integrins, can strongly inhibit curli-mediated internalization. The ability of curli fibers to bind fibronectin can therefore be linked to virulence. PMID- 12076788 TI - Degradation of contrasting pesticides by white rot fungi and its relationship with ligninolytic potential. AB - The capacity of nine species of white rot fungus from a variety of basidiomycete orders to degrade contrasting mono-aromatic pesticides was investigated. There was no relationship between degradation of the dye Poly R-478, a presumptive test for ligninolytic potential, and degradation of the highly available pesticides diuron, metalaxyl, atrazine or terbuthylazine in liquid culture. However, there were significant positive correlations between the rates of degradation of the different pesticides. Greatest degradation of all the pesticides was achieved by Coriolus versicolor, Hypholoma fasciculare and Stereum hirsutum. After 42 days, maximum degradation of diuron, atrazine and terbuthylazine was above 86%, but for metalaxyl less than 44%. When grown in the organic matrix of an on-farm "biobed" pesticide remediation system, relative degradation rates of the highly available pesticides by C. versicolor, H. fasciculare and S. hirsutum showed some differences to those in liquid culture. While H. fasciculare and C. versicolor were able to degrade about a third of the poorly available compound chlorpyrifos in biobed matrix after 42 days, S. hirsutum, which was the most effective degrader of the available pesticides, showed little capacity to degrade the compound. PMID- 12076789 TI - Induction of an outer membrane protein of 78 kDa in Vibrio vulnificus cultured in the presence of desferrioxamine B under iron-limiting conditions. AB - Although Vibrio vulnificus is known to be able to utilize ferrioxamine B as an iron source, its outer membrane receptor remains to be determined. In this study, we found that V. vulnificus expressed a new outer membrane protein of 78 kDa when grown in the presence of desferrioxamine B under iron-limiting conditions. The desferrioxamine B-dependent iron uptake was only observed in bacterial cells expressing this protein. Furthermore, non-denaturing polyacrylamide gel electrophoresis followed by autoradiography of the outer membrane preparation containing the 78-kDa protein preincubated with [(55)Fe]ferrioxamine B provided a single radioactive band in which the 78-kDa outer membrane protein was present as the major component. These lines of evidence suggest that the inducible 78-kDa protein may serve as the cell-surface receptor for ferrioxamine B. PMID- 12076790 TI - Polyhydroxyalkanoate biosynthesis in Pseudomonas pseudoalcaligenes YS1. AB - Pseudomonas pseudoalcaligenes strain YS1 isolated from oil contaminated soil was able to produce polyhydroxybutyrate blended with medium-chain-length polyhydroxyalkanoates (mcl PHA). PHA synthesis genes were cloned from this strain. A fadB (gene for fatty acid degradation) deleted mutant Escherichia coli KM32B (FADB::Tet) was constructed to express the cloned PHA synthesis gene phaC1(Pp) or phaC2(Pp). The fadB deleted mutant KM32B harboring phaC1(Pp) or phaC2(Pp) showed mcl PHA accumulation while the intact E. coli KM32 did not. The results demonstrated that P. pseudoalcaligenes YS1 possessed at least two PHA synthesis pathways; one of them was responsible for production of mcl PHA. PMID- 12076791 TI - Homonucleotide stretches in chromosomal DNA of Campylobacter jejuni display high frequency polymorphism as detected by direct PCR analysis. AB - Homopolymeric nucleotide tracts have been previously identified in the genome sequence of Campylobacter jejuni 11168 [Parkhill et al., Nature 403 (2000) 665 668]. These tracts are believed to regulate contingency genes but as yet no phenotypic variation has been identified associated with many of these genes. To investigate homopolymeric tracts for genes for which there is no observable phenotype, a method was designed to visualise profiles of the various tract lengths directly at the genomic level by means of PCR and denatured polyacrylamide gel electrophoresis. Six of the seven contingency genes investigated displayed variation in the length of the respective homonucleotide tracts. Surprisingly, each contingency gene gave a typical peak profile that represented a conserved size distribution of polymorphic forms. For each gene studied, peak profiles were conserved between strains of C. jejuni. Duplicated genes, containing homonucleotide stretches, displayed locus-specific peak distributions for each gene copy. Contingency genes were polymorphic within single colonies, and the observed complex peak profiles suggested a frequency of slippage several orders of magnitude higher than reported for other organisms. No G7 (or C7) stretch was ever observed, and their absence from the complete genome suggests strong selection against their presence. In view of the predictable outcome of the process leading to these polymorphisms, it is hypothesised that the formation and/or selection of these tracts is not a random process, but is driven by as yet unknown mechanism(s). High-frequency polymorphism of these genes may be a mechanism by which C. jejuni survives selection bottlenecks between opportunities for growth within a host. PMID- 12076792 TI - Passage in mice causes a change in the ability of Salmonella enterica serovar Oranienburg to survive NaCl osmotic stress: resuscitation from the viable but non culturable state. AB - An outbreak caused by dried processed squids contaminated with Salmonella Oranienburg occurred in Japan in 1999. Isolates obtained from the causative food were resistant to NaCl osmotic stress, but isolates from the patients were sensitive to NaCl. Although strains from both sources were almost identical in their virulence in mice, a NaCl-resistant strain from food (Sa9911T) became NaCl sensitive after passage in mice and a NaCl-sensitive strain from one patient (Sa99004) retained NaCl sensitivity after such passage. When dried squid was contaminated experimentally with both strains during processing, only Sa9911T was recovered directly from the final product. Nevertheless, the viability of the Sa99004 cells was over 90% found by fluorescent staining. We suggested that Sa99004 might become viable but non-culturable (VNC) by NaCl stress. This hypothesis was confirmed by resuscitation by efficient enrichment. We concluded that VNC S. Oranienburg would be potentially dangerous contaminants of NaCl preserved foods and that measures to ensure their detection should be taken at the time of food inspection. PMID- 12076793 TI - The copy number of the catabolic plasmid pJP4 affects growth of Ralstonia eutropha JMP134 (pJP4) on 3-chlorobenzoate. AB - Ralstonia eutropha JMP134 (pJP4) grows on 3-chlorobenzoate (3-CB) or 2,4 dichlorophenoxyacetate (2,4-D). The copy number of chlorocatechol genes has been observed to be important for allowing growth of bacterial strains on chloroaromatic compounds. Despite the fact that two functional chlorocatechol degradation tfd gene clusters are harbored on plasmid pJP4, a single copy of the region comprising all tfd genes in strain JMP134-F was insufficient to allow growth on 3-CB, whereas growth on 2,4-D was only slightly retarded compared to the wild-type strain. Using competitive PCR, approximately five copies of pJP4 per genome were observed to be present in the wild-type strain, whereas only one copy of pJP4 was present per chromosome in strain JMP134-F. Therefore, several copies of pJP4 per chromosome are required for full expression of the tfd-encoded growth abilities in the wild-type R. eutropha strain. PMID- 12076794 TI - Role of the Pseudomonas aeruginosa las and rhl quorum-sensing systems in rhlI regulation. AB - In Pseudomonas aeruginosa the LasR-LasI and RhlR-RhlI quorum-sensing (QS) systems control expression of numerous virulence genes in a population density-dependent fashion. In this study, we investigated regulation of the autoinducer synthase gene rhlI, which is responsible for C(4)-HSL signal production. Primer extension analysis was used to map the rhlI transcriptional start site and an upstream regulatory region was identified. Expression studies revealed that (i) this regulatory region is important for rhlI expression and (ii) although the rhl QS system will induce rhlI, las is the dominant regulator. Furthermore, we found that control of rhlI in Escherichia coli is markedly different than in P. aeruginosa. PMID- 12076795 TI - Effect of culture conditions on Escherichia coli O157:H7-mediated attaching effacing lesions in a bovine large intestinal mucosal explant model. AB - The effects of culture conditions on the extent of Escherichia coli O157:H7 attaching-effacing (A/E) adherence in an adult bovine large intestinal mucosal explant model were assessed by three different morphometric methods. Measurement of the percent of tissue sections with A/E adherence and the number of foci of A/E adherence mm(-1) of surface epithelium was more sensitive than measurement of the percent of surface epithelium with A/E adherent bacteria for detection of treatment effects. Culture of bacterial inoculum in tryptic soy broth, incubation of explants in 5% CO(2), and rocking of explants on a platform rocker at 18 cycles min(-1) provided optimal conditions for A/E adherence. In future studies, the model may be used for preliminary testing of intervention strategies aimed at reduction of E. coli O157:H7 intestinal colonization of cattle. PMID- 12076796 TI - Molecular characterization of phosphoglycerate mutase in archaea. AB - The interconversion of 3-phosphoglycerate and 2-phosphoglycerate during glycolysis and gluconeogenesis is catalyzed by phosphoglycerate mutase (PGM). In bacteria and eukaryotes two structurally distinct enzymes have been found, a cofactor-dependent and a cofactor-independent (iPGM) type. Sequence analysis of archaeal genomes did not find PGMs of either kind, but identified a new family of proteins, distantly related to iPGMs. In this study, these predicted archaeal PGMs from Pyrococcus furiosus and Methanococcus jannaschii have been functionally produced in Escherichia coli, and characterization of the purified proteins has confirmed that they are iPGMs. Analysis of the available microbial genomes indicates that this new type of iPGM is widely distributed among archaea and also encoded in several bacteria. In addition, as has been demonstrated in certain bacteria, some archaea appear to possess an alternative, cofactor-dependent PGM. PMID- 12076797 TI - Cloning, expression and characterization of L-arabinose isomerase from Thermotoga neapolitana: bioconversion of D-galactose to D-tagatose using the enzyme. AB - Gene araA encoding an L-arabinose isomerase (AraA) from the hyperthermophile, Thermotoga neapolitana 5068 was cloned, sequenced, and expressed in Escherichia coli. The gene encoded a polypeptide of 496 residues with a calculated molecular mass of 56677 Da. The deduced amino acid sequence has 94.8% identical amino acids compared with the residues in a putative L-arabinose isomerase of Thermotoga maritima. The recombinant enzyme expressed in E. coli was purified to homogeneity by heat treatment, ion exchange chromatography and gel filtration. The thermophilic enzyme had a maximum activity of L-arabinose isomerization and D galactose isomerization at 85 degrees C, and required divalent cations such as Co(2+) and Mn(2+) for its activity and thermostability. The apparent K(m) values of the enzyme for L-arabinose and D-galactose were 116 mM (v(max), 119 micromol min(-1) mg(-1)) and 250 mM (v(max), 14.3 micromol min(-1) mg(-1)), respectively, that were determined in the presence of both 1 mM Co(2+) and 1 mM Mn(2+). A 68% conversion of D-galactose to D-tagatose was obtained using the recombinant enzyme at the isomerization temperature of 80 degrees C. PMID- 12076798 TI - Regulation of the Legionella mip-promotor during infection of human monocytes. AB - The opportunistic pathogen Legionella pneumophila, the etiologic agent of Legionnaires disease, is able to invade and multiply intracellularly in human macrophages. This process is controlled by several bacterial virulence factors. As recently demonstrated, one of these virulence factors, the macrophage infectivity potentiator (Mip) protein, is important for invasion and proper intracellular establishment of L. pneumophila in macrophages and protozoa. Knockout mutants devoid of a functional mip-gene enter host cells much less effectively but intracellular replication is not affected. Using a P(mip)-green fluorescent protein reporter construct in L. pneumophila substrain Corby, P(mip) was recently shown to be constitutively active in replicating bacteria. A stringent regulation during the infection process could not be observed, neither in intracellular nor in BYE broth-grown bacteria. For enhanced temporal and quantitative resolution, we examined the activity of mip on RNA level in order to detect short transient regulatory events. Our results show that P(mip) of L. pneumophila is temporarily repressed directly after invasion of the monocytic human cell line MonoMac 6 and regains activity after 24 h of intracellular replication. PMID- 12076799 TI - The membrane-associated F(0)F(1) ATPase is essential for the viability of Streptococcus pneumoniae. AB - Genetic studies aimed at eliminating expression of the atp operon (F(0)F(1) H(+) ATPase) of Streptococcus pneumoniae by genetic disruption of atpC, the first gene of the operon, with a chloramphenicol-resistance cassette were performed. Resistant transformants were obtained only when the recipient strain had a duplication of atpC, recombination occurring in such a way that transcription of the operon from its own promoter was allowed. These results imply that the atp operon is essential for the viability of the cells. PMID- 12076801 TI - Characterisation of the QM gene of Trypanosoma brucei. AB - The QM protein has been reported to have roles in both tumour suppression and transcription factor regulation in vertebrate cells, and in ribosome stability in both yeast and mammals. The present study isolated the QM gene of Trypanosoma brucei and determined its sequence. Alignment with QM sequences from Saccharomyces cerevisiae, Arabidopsis thaliana, Drosophila melanogaster and Homo sapiens revealed greater than 60% identity. Southern blot analysis revealed multiple copies of QM within the trypanosome genome. An epitope tag was inserted into the C-terminus of the T. brucei QM and the protein expressed under inducible control in procyclic form trypanosomes. Immune fluorescence microscopy revealed co-localisation with the GPI:protein transamidase component, GPI8, a distribution indicative of ribosome association with the rough endoplasmic reticulum. PMID- 12076800 TI - Anaerobic degradation of protocatechuate (3,4-dihydroxybenzoate) by Thauera aromatica strain AR-1. AB - The denitrifying bacterium Thauera aromatica strain AR-1 grows anaerobically with protocatechuate (3,4-dihydroxybenzoate (DHB)) as sole energy and carbon source. This bacterium harbors two distinct pathways for degradation of aromatic compounds, the benzoyl-coenzyme A (CoA) pathway for benzoate degradation and the hydroxyhydroquinone (HHQ) pathway for degradation of 3,5-DHB. In order to elucidate whether protocatechuate is degraded via the benzoyl-CoA or the HHQ pathway, induction experiments were carried out. Dense suspensions of cells grown on protocatechuate or benzoate readily degraded benzoate and protocatechuate but not 3,5-DHB. Dense suspensions of 3,5-DHB-grown cells degraded 3,4- and 3,5-DHB at similar rates, but benzoate was not degraded. 3,5-DHB hydroxylating activity was found only in cells grown with this substrate. HHQ dehydrogenase activity was found in extracts of cells grown with 3,5-DHB and at a low rate also in protocatechuate-grown cells, but not in extracts of cells grown with benzoate. Activities of protocatechuyl-CoA synthetase and protocatechuyl-CoA reductase leading to 3-hydroxybenzoyl-CoA were found in extracts of cells grown with protocatechuate. There was no repression of the HHQ pathway by the presence of protocatechuate, unlike by degradation of benzoate. We conclude that protocatechuate is not degraded via the HHQ pathway because there was no evidence of a hydroxylation reaction involved in this process. Instead, our results strongly suggest that protocatechuate is degraded via a pathway which connects to the benzoyl-CoA route of degradation. PMID- 12076802 TI - Construction and intergeneric conjugative transfer of a pSG5-based cosmid vector from Escherichia coli to the polyisoprene rubber degrading strain Micromonospora aurantiaca W2b. AB - A non-rubber degrading mutant of the polyisoprene rubber degrading bacterium Micromonospora aurantiaca W2b lacking the capability to form halos on latex overlay agar plates was isolated after N-methyl-N-nitro-N-nitrosoguanidine mutagenesis. A 10.3-kb shuttle cosmid vector pGM446 was constructed from the Streptomyces cloning vectors pGM160 and pOJ446. This vector was transferred by conjugation from Escherichia coli to M. aurantiaca W2b. The frequency of formation of exconjugants with pGM446 was 3.6 x 10(-3). This vector could be useful for shotgun cloning of genes into the non-rubber degrading mutant L1 from M. aurantiaca W2b. PMID- 12076803 TI - Cloning, expression, and functional characterization of the Mycobacterium tuberculosis secA gene. AB - To better understand the protein secretion mechanisms involved in the growth and pathogenesis of Mycobacterium tuberculosis, we examined the secA gene from M. tuberculosis (tbsecA; cosmid sequence accession No. z95121.gb_ba). We generated plasmids containing the full-length tbsecA gene or a fusion containing the 5' sequence from the M. tuberculosis secA gene and the remainder from the Escherichia coli secA gene and evaluated the ability of each construct to complement the defective SecA protein in E. coli MM52ts when grown at the non permissive temperature. The full-length tbsecA gene was unable to compensate for the temperature-sensitive defect, whereas E. coli MM52ts that has been transformed with plasmid pMF8TB226 containing a chimeric secA gene was able to grow at 42 degrees C. This work confirms that the topography of SecA and its ATP binding sites are highly conserved, whereas its membrane insertion domains are species specific. PMID- 12076804 TI - Transformation of the edible mushroom Pleurotus ostreatus by particle bombardment. AB - Uracil auxotroph of Pleurotus ostreatus was transformed to prototrophy by means of particle bombardment. Five transformants were obtained under three conditions differing in the two parameters of target distance and helium pressure. The transformation frequency was one transformant per microg of DNA. In the transformants, plasmid DNAs were integrated into the genomic DNA and stably maintained. This is the first report on transformation of P. ostreatus by particle bombardment. PMID- 12076805 TI - Monitoring the identity of bacteria using their intrinsic fluorescence. AB - Three different fluorescence spectra were recorded following excitation at 250 nm (aromatic amino acids+nucleic acids), 270 nm (tryptophan residues) and 316 nm (NADH) for 25 strains of bacteria in dilute suspensions. Evaluation of the spectra using principal component analysis and hierarchical clustering showed a good reproducibility from culture to culture and a good discrimination of the bacteria. Applying the method of Mahalanobis distances to the spectra of lactobacilli species recorded following excitation at 250 nm, a good classification was observed for 100% and 81% of calibration and validation groups, respectively. The developed method allows the discrimination and identification of the investigated bacteria at the genus, species and strain levels. PMID- 12076806 TI - Inhibitory effect of sulfur dioxide and other stress compounds in wine on the ATPase activity of Oenococcus oeni. AB - Malolactic fermentation (MLF) is carried out by Oenococcus oeni under very harsh conditions. This paper shows that stress compounds in wine such as SO(2), fatty acids and copper have an inhibitory effect on cell growth and MLF duration, and relates this effect to an inhibition of ATPase activity. Of the stress compounds, SO(2) and dodecanoic acid had the strongest effect, decreasing the ATPase specific activity to 37% and 58%, respectively. It can be concluded that ATPase is a good indicator of the physiological state of the cells and their ability to lead MLF. PMID- 12076807 TI - Temperature-dependent translation of leaderless and canonical mRNAs in Escherichia coli. AB - Leaderless mRNAs beginning with a 5'-terminal start codon occur in all biological systems. In this work, we have studied the comparative translational efficiency of leaderless and leadered mRNAs as a function of temperature by in vitro translation competition assays with Escherichia coli extracts. At low temperature (25 degrees C) leaderless mRNAs were found to be translated comparatively better than mRNAs containing an internal canonical ribosome binding site, whereas at high temperature (42 degrees C) the translational efficiency of canonical mRNAs is by far superior to that of leaderless mRNA. The inverse correlation between temperature and translational efficiency characteristic for the two mRNA classes was attributed to structural features of the mRNA(s) and to the reduced stability of the translation initiation complex formed at a 5'-terminal start codon at elevated temperature. PMID- 12076808 TI - Alpha-proteobacteria cultivated from marine sponges display branching rod morphology. AB - Most isolates recovered from marine environments are Gram-negative proteobacteria, even with the use of various media and media additions to enhance recoverability. Cultivation studies with two genera of deep-water sponges yielded nine isolates that demonstrated bulbous branching rod morphology, which is usually associated with microorganisms staining Gram-positive. Gram reactions indicated that the isolates were Gram-negative, which was confirmed by partial 16S rDNA sequencing. All nine isolates were shown to be alpha-proteobacteria most closely related to other alpha-proteobacteria isolated from various sponges. PMID- 12076809 TI - Use of subtractive hybridization for comprehensive surveys of prokaryotic genome differences. AB - Comparative bacterial genomics shows that even different isolates of the same bacterial species can vary significantly in gene content. An effective means to survey differences across whole genomes would be highly advantageous for understanding this variation. Here we show that suppression subtractive hybridization (SSH) provides high, representative coverage of regions that differ between similar genomes. Using Helicobacter pylori strains 26695 and J99 as a model, SSH identified approximately 95% of the unique open reading frames in each strain, showing that the approach is effective. Furthermore, combining data from parallel SSH experiments using different restriction enzymes significantly increased coverage compared to using a single enzyme. These results suggest a powerful approach for assessing genome differences among closely related strains when one member of the group has been completely sequenced. PMID- 12076810 TI - Photoinduced bactericidal activity against Pseudomonas aeruginosa by TiO(2) based thin films. AB - The photoinduced bactericidal capacity of TiO(2) based films was evaluated, using as model organism Pseudomonas aeruginosa. Thin films were obtained by spray pyrolysis; they included undoped, Cu doped, and Al doped TiO(2). Scanning electron microscopy was used to observe the final effect of the irradiated films upon the bacteria. Depending on the composition and characteristics of the films, quantitative experiments show that bacterial inhibition varies between 28 and 96%. The order of magnitude of the average quantum yield of the films was determined between 10(-9) and 10(-11) inhibited bacterial per photon. PMID- 12076811 TI - Osmoprotection by carnitine in a Listeria monocytogenes mutant lacking the OpuC transporter: evidence for a low affinity carnitine uptake system. AB - A deletion mutant of Listeria monocytogenes lacking OpuC, an ABC transporter responsible for the uptake of the compatible solute carnitine, was constructed and carnitine transport assays confirmed that carnitine transport was defective in this mutant. However, the mutant retained the ability to derive osmoprotection from carnitine, suggesting the presence of a second uptake system for this compatible solute. Measurement of intracellular carnitine pools during balanced growth confirmed that the opuC mutant accumulated high levels of carnitine. These pools were only achieved in the mutant when high levels (1 mM) of carnitine were present extracellularly. When a lower level (100 microM) was supplied in the medium the mutant failed to accumulate a substantial intracellular pool and failed to derive osmoprotection from carnitine. These data suggest the presence of a second low affinity carnitine uptake system in this osmotolerant pathogen. PMID- 12076812 TI - Diversity of culturable bacterial populations associated to Tuber borchii ectomycorrhizas and their activity on T. borchii mycelial growth. AB - Isolation and physiological and molecular characterisation of culturable bacterial strains belonging to actinomycetes, pseudomonads and aerobic spore forming bacteria were carried out on mycorrhizal root tips of Quercus robur var. peduncolata infected by Tuber borchii. Cellular density of the three bacterial groups in ectomycorrhizal root tips was estimated to be 1.3+/-0.11 x 10(6) cfu g( 1) dry weight for total heterotrophic bacteria and 1.08+/-0.6 x 10(5) (mean+/ S.E.), 1.3+/-0.3 x 10(5) and 1.4+/-0.2 x 10(5) cfu g(-1) dry weight for pseudomonads, actinomycetes and spore-forming bacteria respectively. Identification of pseudomonads by the Biolog system indicated, besides the most represented species Pseudomonas fluorescens (biotypes B, F and G), the occurrence of strains belonging to Pseudomonas corrugata. Amplified ribosomal DNA restriction analysis of actinomycetes and spore formers revealed at least three and six different groups of patterns, respectively. Many bacterial isolates were able to induce variations in growth rates of T. borchii mycelium; among these, 101 strains showed antifungal activity, whereas 17 isolates, belonging to spore formers, were able to increase mycelial growth up to 78% when compared to uninoculated mycelial growth. The potential role of these populations in the development and establishment of mycorrhizas is discussed. PMID- 12076813 TI - In vitro anti-Neisseria gonorrhoeae activity of Terminalia macroptera leaves. AB - We used the agar dilution method to evaluate the antibacterial effect of Terminalia macroptera leaf (Tml) extract against nine reference and clinical Neisseria gonorrhoeae strains, including penicillin- and tetracycline-resistant and -susceptible strains. Tml possesses anti-N. gonorrhoeae activity against all of the strains and the minimum inhibitory concentrations (MIC) were between 100 and 200 microg ml(-1). We then used a liquid-liquid partition method to divide the Tml extract into five fractions and determined the anti-N. gonorrhoeae activity of each of the fractions. All of the fractions showed antibacterial activity. The most active one was identified as the diethyl ether fraction and had MIC values of between 25 and 50 microg ml(-1) against all of the strains. PMID- 12076814 TI - Survival of spumavirus, a primate retrovirus, in laboratory media and water. AB - The persistence of a previously characterized spumavirus strain (strain SV-522) was investigated utilizing various laboratory media and waters, including Eagle's minimal essential medium (EMEM) plus 0% fetal bovine serum (EMEM-0%), EMEM-2%, EMEM-10%, Chlamydia transport medium (CTM), phosphate-buffered saline, distilled, estuarine, and marine water, human serum, and the germicides, ethyl alcohol (70%) and sodium hypochlorite (10%). Experiments were performed at 4 degrees C and/or 23 degrees C. Infectivity endpoints were determined in stock aliquots upon initiation of testing and then after 3, 5, 7, and 10 days. The virus was reisolated from all diluents after 5 days at 23 degrees C and in EMEM-10% after 7 days. The virus was detected in CTM, EMEM-2%, EMEM-10%, and estuarine and marine waters after 7 days at 4 degrees C. Differences in the persistence of the virus may be ascribed to temperature and organic load. Water ionic strengths (e.g., estuarine vs. marine water) had no effect on modifying persistence of viral particles. Infectivity of spumavirus was undetectable after 30 s in 70% ethanol or 10% sodium hypochlorite. After 30 min at 23 degrees C, spumavirus infectivity in normal but not heat-inactivated human serum increased by almost 100-fold. Persistence of infectivity of primate spumavirus after 7 days in media and waters, and the agent's infectious potential in the human host, emphasize a need for cautious recognition during the manipulation of primate cells/organs and in the handling of primates themselves. PMID- 12076815 TI - The substrate-binding protein TeaA of the osmoregulated ectoine transporter TeaABC from Halomonas elongata: purification and characterization of recombinant TeaA. AB - TeaABC is a novel, osmoregulated secondary transport system for compatible solutes found in the halophilic proteobacterium Halomonas elongata DSM 2581(T). Sequence comparisons have confirmed that TeaABC belongs to the family of TRAP transporters and as such it consists of two putative transmembrane proteins (TeaB, TeaC) and a putative periplasmic substrate-binding protein (TeaA). TeaABC is the only osmoregulated transporter for ectoines found in H. elongata. By overexpressing TeaA-(6)His tag in Escherichia coli we demonstrated that TeaA is processed as predicted and exported to the periplasm. Furthermore, it was proven that TeaA is indeed a periplasmic ectoine-binding protein. The functionality of the purified protein as an ectoine-binding protein was tested employing a modified binding assay. PMID- 12076816 TI - AbrB is a regulator of the sigma(W) regulon in Bacillus subtilis. AB - The Bacillus subtilis global regulator AbrB was found to negatively control expression of sigW and genes of the sigma(W) regulon. AbrB bound to DNA regions in the autoregulatory sigW promoter and to some, but not all, of the other sigma(W)-dependent promoters in B. subtilis. Defects in antibiotic resistance properties caused by spo0A mutations are at least partially correlated with AbrB repression of the sigma(W) regulon. PMID- 12076817 TI - Prevalence of granulocytic Ehrlichiae in Ixodes ricinus ticks in Middle Germany (Thuringia) detected by PCR and sequencing of a 16S ribosomal DNA fragment. AB - A total of 305 Ixodes ricinus ticks (243 nymphs and 62 adults) were collected from three different regions of Thuringia in Middle Germany which are known to be endemic for Borrelia burgdorferi. Our aim was to investigate the carrier rate of ticks for granulocytic Ehrlichia species. The presence of ehrlichial 16S ribosomal DNA was investigated by polymerase chain reaction. Using primers specific for the Ehrlichia phagocytophila group PCR fragments of 151 bp and 943 bp, respectively, were produced in positive samples. Adult ticks showed a significantly higher infection rate (4/62; 6.5%) compared to nymphs (3/243; 1.2%). Prevalence rates varied between 0 and 3.8% regarding the different areas under investigation. The nucleotide sequences showed high similarity (between 97.5% and 99% identity) to the known sequences of the three E. phagocytophila group members HGE agent, E. phagocytophila and Ehrlichia equi. The sequence data did not allow a final classification to a particular member of this group. PMID- 12076819 TI - Bacterial adhesion to sulcular epithelium in periodontitis. AB - The purpose of this study was to investigate, by electron microscopy, the type of bacterial attachment to the sulcular epithelium in periodontitis. Gingiva biopsies were observed in a transmission electron microscope using cytochemical staining with ruthenium red for glycocalyx visualisation. In addition, subgingival plaque samples and biopsies from the sulcular epithelium in periodontitis from the patients were estimated microbiologically. Aerobic bacteria only were estimated in the subgingival plaque and both aerobic and anaerobic bacteria in the gingival biopsies. No bacterial internalisation could be observed. Fimbria-mediated adhesion as the only type of bacterial attachment and a large diversity of bacterial glycocalyces were detected. As the fimbrial adhesins of putative periodontal pathogens are able in vitro to induce inflammation and bone resorption via stimulation of the proinflammatory cytokine production, the demonstrated fimbrial adhesins suggest the significant role of bacterial adhesion to sulcular epithelium in periodontitis. PMID- 12076818 TI - Molecular analysis of the dormancy response in Mycobacterium smegmatis: expression analysis of genes encoding the DevR-DevS two-component system, Rv3134c and chaperone alpha-crystallin homologues. AB - Mycobacteria adapt to a decrease in oxygen tension by entry into a non replicative persistent phase. It was shown earlier that the two-component system, DevR-DevS, was induced in Mycobacterium tuberculosis and Mycobacterium bovis BCG cultures during hypoxia, suggesting that it may play a regulatory role in their adaptation to oxygen limitation. The presence of a homologous genetic system in Mycobacterium smegmatis was predicted by scanning its unfinished genome sequence with devR and devS genes of M. tuberculosis. Rv3134c, which is cotranscribed with devR-devS in M. tuberculosis, was also present in M. smegmatis at a similar location upstream from devR. The expression of all three genes was induced at the RNA and protein levels in M. smegmatis cultures grown under microaerobic and anaerobic conditions. The M. smegmatis genome also contained the hspX gene, encoding chaperone alpha-crystallin, Acr, that was induced during hypoxia. The similarity in sequences and hypoxia-responsive behaviour of devR-devS, Rv3134c and hspX genes in M. smegmatis and M. tuberculosis suggests that the molecular mechanisms involved in the dormancy response are likely conserved in these two species. M. smegmatis could therefore serve as a useful model for the delineation of the hypoxia response in general and DevR-DevS regulated pathways in particular. PMID- 12076820 TI - Characterization of Bacillus strains from apple and pear trees in South Africa antagonistic to Erwinia amylovora. AB - In order to find reasons for the absence of fire blight in most countries of the Southern hemisphere, bark samples from apple and pear trees in orchards of the Western Cape region in South Africa were extracted for bacteria which could be antagonistic to Erwinia amylovora. Screening was done in the late growth season and mainly Gram-positive bacteria were isolated. Approximately half of them produced growth inhibition zones on a lawn of E. amylovora. Most isolates were classified as Bacillus megaterium by microbiological assays and in API 50 test systems. They were visualized in the light microscope as non-motile large rods. These strains may not be responsible for the absence of fire blight in orchards, but they may indicate unfavourable climatic conditions for Gram-negative bacteria including E. amylovora. They may reduce the ability of E. amylovora to establish fire blight and could also be useful for application in biological disease control. PMID- 12076821 TI - Regulatory role of external calcium on Pythium porphyrae (Oomycota) zoospore release, development and infection in causing red rot disease of Porphyra yezoensis (Rhodophyta). AB - Formation of zoosporangia and cleavage of zoosporangial cytoplasm to zoospores in Pythium porphyrae is absolutely dependent on extracellular calcium. Calcium ion could be substituted neither by monovalent nor divalent cations tested. Increased concentrations of extracellular calcium did not affect the release of zoospores from zoosporangia but inhibited the zoospore motility. Chelating calcium ion by EGTA has decreased the ability of encysted zoospores to germinate and form appressoria. The increased external calcium-ion concentration has decreased the infectivity of Porphyra yezoensis thalli in a linear fashion apparently indicating a role of calcium in the signaling mechanism. PMID- 12076822 TI - Sec-dependent and Sec-independent translocation of haloacid dehalogenase Chd1 of Burkholderia cepacia MBA4 in Escherichia coli. AB - 2-Haloacid dehalogenases are hydrolytic enzymes that cleave the halogen-carbon bond(s) in haloalkanoic acids. We have previously isolated a cryptic haloacid dehalogenase gene from Burkholderia cepacia MBA4 and expressed it in Escherichia coli. This recombinant protein is unusual in having a long leader sequence, a property of periplasmic enzymes. In this paper, we report the functional role of this leader sequence. Western blot analyses showed that Chd1 is translocated to the periplasm. The results on the expression of Chd1 in the presence of sodium azide suggested the cleavage of the leader to be Sec-dependent. Chimeras of Chd1 and green fluorescent protein demonstrated that the leader sequence is fully functional in translocating the fusion protein to the periplasm. The expression of the chimeras in Sec mutants supported the Sec-dependent translocation. Surprisingly, recombinant Chd1 and a chimera with no leader sequence were also found in the periplasm. PMID- 12076824 TI - A fitness cost for Turnip mosaic virus to overcome host resistance. AB - The relative fitness of the Turnip mosaic virus (TuMV) isolate UK 1 was compared with that of two other wildtype isolates CZE 1 and CDN 1. The isolates CZE 1 and CDN 1 are able to overcome the effect of the resistance gene TuRB01 and at least three other resistance sources that are effective against UK 1. Comparisons were also made between the fitness of UK 1 and a recombinant virus with a single nucleotide change (v35Tunos +5570 A>G) conferring the ability to overcome TuRB01 resistance. Co-inoculation experiments were carried out where pairs of isolates were serially passaged over 5 months in a plant line possessing no known resistance genes in order to examine the relative fitness of the isolates. In each case, UK 1 dominated the mixture in the susceptible host background. It out competed CZE 1 and v35Tunos +5570 A>G within four passages, and CDN 1 after one passage. The greater fitness of UK 1 suggests that there may be a fitness cost to TuMV overcoming resistance genes of brassica crops. This may shed some light on the frequency of naturally occurring isolates, in that pathotype 1 isolates are found much more frequently than isolates of other pathotypes. Implications for the deployment of TuRB01 are discussed. PMID- 12076823 TI - Escherichia coli genes involved in resistance to pyrazinoic acid, the active component of the tuberculosis drug pyrazinamide. AB - The natural resistance of Escherichia coli to pyrazinoic acid (POA), the active derivative of pyrazinamide, was investigated. The TolC mutant was found to be more susceptible to POA and other weak acids than the wild-type strain. Mutation in EmrB but not AcrB efflux protein slightly increased POA susceptibility. Two transposon mutants with increased susceptibility to POA were found to harbor mutations in acnA encoding aconitase-1 and ygiY encoding a putative two-component sensor protein. Complementation of the AcnA and YgiY mutants conferred resistance to POA, whereas the complemented TolC mutant became resistant to POA and other weak acids. PMID- 12076825 TI - Cell-binding properties of glycoprotein B of Aujeszky's disease virus. AB - The glycoprotein B (gB) of Aujeszky's disease virus (ADV) has a role in virus entry and cell-to-cell spread. In this report we examined the cell-binding properties of native ADV gB purified from the virus envelope by affinity chromatography. The binding of gB to the surface of susceptible cells BHK-21 and MDBK was specific, dose-dependent, and nearly saturable, which is characteristic of conventional receptor-ligand interactions. The purified gB was shown to specifically bind to immobilised heparin. The addition of soluble exogenous heparin and heparinase treatment of cells inhibited the binding of gB to the cells. Cell-associated gB could also be dissociated from the cells by soluble heparin. The results indicated that ADV gB binds specifically to cellular heparan sulphate. The binding of gB to cells inhibited the attachment of virus to cells and thus the formation of viral plaques. The results suggest that ADV gB may have a function in the initial attachment of ADV to the surface of susceptible cells. PMID- 12076826 TI - Enhanced protection against HSV lethal challenges in mice by immunization with a combined HSV-1 glycoprotein B:H:L gene DNAs. AB - The effectiveness of a cocktailed HSV-1 three-glycoprotein B, H, and L gene vaccine in comparison to individual glycoprotein gene vaccines was studied with regard to protecting against the HSV-1 infection. Three glycoprotein gene recombinant DNA vaccines, which produced the corresponding glycoproteins in Vero cells, were constructed using a CMV promoter. The cocktailed DNA vaccines were prepared by combining all three genes. The titers of neurtalizing antibody following the immunization of the five vaccines were KOS(1/1024)>B:H:L=B(1/512)>H:L(1/64)>H(1/16) genes. The mice, which were immunized with L gene alone failed to induce enough neutralizing antibody. The CTL activity was rated as KOS (95%)>B:H:L (80%)>B(60%)>H:L(50%)> H (35%) gene vaccines at an E:T ratio of 50:1. The H gene alone or L gene vaccine alone induced little CTL activity. The protection rates of the DNA-vaccinated mice against the lethal intraperitoneal (i.p.) or i.m challenges were shown as KOS>B:H:L>B>H:L>H gene vaccines, and the protection activity depended on the lethal dosage of the challenging virus, which are inversely proportional to each other. Compared with the mice, which were vaccinated with individual DNA vaccines, the mice, which were vaccinated with the cocktailed three-gene vaccine, were shown to be better protected against the lethal challenging doses. It can be concluded that vaccination with the cocktailed three gene vaccines is more effective in protecting mice from the viral challenge and the protection rate varies inversely with the amount of lethal challenging dose used, although all DNA vaccines failed to block the latent infection in sensory nerves. PMID- 12076827 TI - Recombinant Sendai viruses expressing different levels of a foreign reporter gene. AB - Sendai virus (SeV) is an enveloped virus with a nonsegmented negative strand RNA genome. The recovery of infectious virus from cDNA and generation of recombinant SeV carrying a foreign gene at the promoter proximal position has been demonstrated. In this study, we constructed a series of recombinant SeVs carrying a reporter human secreted alkaline phosphatase (SEAP) gene at each viral gene junction or the 5' distal end in order to measure the expression level of the foreign gene. We demonstrated that there was a gradient in the reporter gene expression level that depended on location, due to the polarity of transcription. In contrast, the growth and final titers of these recombinant viruses showed an opposite gradient to the foreign gene expression level. This suggests the potential for matching therapeutic gene expression level to individual therapy programs by changing the position of the foreign gene when SeVs are used as vectors for human gene therapy. PMID- 12076828 TI - Interactions between human cytomegalovirus helicase-primase proteins. AB - The human cytomegalovirus (HCMV) UL70, UL102, and UL105 genes are predicted to encode essential proteins that assemble the replicative helicase-primase complex based on sequence and genome position similarities to putative herpes simplex virus type 1 (HSV-1) counterparts. Consistent with this prediction, they are required for transient complementation of DNA synthesis. However, little is known about their physical interactions and biochemical activities, primarily because of their restricted expression in HCMV-infected cells. To look for assembly of the predicted complexes, we prepared rabbit polyclonal antisera and used Semliki Forest Virus (SFV) vectors to express untagged and glutathione-S-transferase (GST)-tagged UL70, UL102 and UL105 proteins. The UL70 and UL105 proteins co purified with the GST-tagged UL102 protein from triply-infected baby hamster kidney cells (BHK-21), and pUL70, but not pUL105, co-purified with pGST-UL102 from dually infected BHK-21 cells. In immunoprecipitation experiments with untagged SFV-expressed proteins, pUL70 or pUL105 coprecipitated with pUL102, pUL102 or pUL70 co-precipitated with pUL105; and pUL102 or pUL105 coprecipitated with pUL70. Comparison of the GST-pull down and immunoprecipitation experiments suggested that the amino-terminal GST-tag interfered with certain pairwise interactions. These results support the prediction that the HCMV helicase-primase proteins assemble a three-protein heteromeric complex, and show that each protein contacts both partners. PMID- 12076829 TI - Reevaluation of a North India isolate of hepatitis E virus based on the full length genomic sequence obtained following long RT-PCR. AB - The genomic cloning and sequence of hepatitis E virus (HEV) from an epidemic in North India is reported. We describe here a simple method wherein the viral RNA was reverse transcribed and then amplified in a single step using an extra long polymerase chain reaction procedure. The full genome nucleotide sequence of this HEV isolate (called Yam-67) was made up of 7191 nucleotides, excepting the poly(A) tail and had three open reading frames: ORF1 coding for 1693 amino acids (aa), ORF2 coding for 659 aa and ORF3 coding for 122 aa. This North Indian isolate of HEV showed close sequence homology to other HEV isolates from India and Asia, but was distant from the Chinese genotype 4, Japanese, Mexican and US isolates. There is no indication from sequence analysis that this may be an atypical strain of HEV, as reported earlier. PMID- 12076830 TI - Analysis of herpes simplex virus entering into cells of oral origin. AB - The entry of herpes simplex virus (HSV) into an oral epithelial cell line, primary normal human oral keratinocytes (NHOK) and gingival fibroblasts (GF) was examined. Infection of these cells by HSV-1 and HSV-2 was blocked by heparin. Further examination indicated that heparin reduced viral attachment but not penetration. Moreover, neomycin inhibited HSV-1 infection more effectively than HSV-2 infection in GF, but not in NHOK. In conclusion, our results elucidated some aspects of the HSV entry process into oral cells and revealed some differences in HSV entering into NHOK and GF. PMID- 12076831 TI - Co-adaptation between cassava mosaic geminiviruses and their local vector populations. AB - Four cassava mosaic geminivirus (CMG) isolates; African cassava mosaic virus from Namulonge, Uganda (ACMV-[Nam]), East African cassava mosaic virus from Mtwara, Tanzania (EACMV-[Mtw]), EACMV-Uganda from Namulonge (EACMV-UG[Nam]) and Indian cassava mosaic virus from Trivandrum, India (ICMV-[Tri]) were compared for their ability to be transmitted by four colonies of cassava whitefly, Bemisia tabaci (Gennadius), collected from Namulonge (NAM), Mtwara (MTW), Kumasi (KUM) and Trivandrum (TRI). With 20 adult whiteflies per test plant, the CMGs from Africa were transmitted by all three of the African B. tabaci populations to 60-79% of the target plants. Indian cassava B. tabaci transmitted ICMV-[Tri] to 89% of the target plants. In contrast, Indian cassava B. tabaci transmitted EACMV-[Mtw] and Tanzanian cassava B. tabaci transmitted ICMV-[Tri] less than a tenth as often, even when using 50 adults per plant and with increased acquisition and inoculation access periods. The complete coat protein genes of the CMGs had sequences typical of their source viruses, the major differences occurring between those originating from India and Africa. Symptom severity of the CMGs was quantified precisely by both visual assessment and image analysis techniques. EACMV-[Mtw] and ACMV-[Nam] were the most and least damaging isolates with 15.4 and 10.0% of the leaf area diseased, respectively. In these tests the transmission frequency was not linked to symptom severity in the source plants. These data support the hypothesis that virus-vector co-adaptation exists in the cassava mosaic disease (CMD) pathosystem and the results are discussed in relation to CMD epidemiology. PMID- 12076833 TI - Transactivation of the human endogenous retrovirus K long terminal repeat by herpes simplex virus type 1 immediate early protein 0. AB - We found that LTR-directed transcription of the human endogenous retrovirus K can be induced by HSV-1 infection. The effect was mediated by the action of a HSV-1 immediate early protein, ICP0 and required the AP-1 binding site present on the HERV-K LTR. In addition, ICP0 could up-regulate AP-1 activity, suggesting that ICP0 increases transcription of HERV-K through AP-1 site. This effect might be important to understand both HERV-K- and HSV-1-mediated pathogenesis because HERV K LTR represents an important class of retrotranspositional mutagens and also could provide a new regulatory element for the linked DNA sequences. PMID- 12076834 TI - Genetic heterogeneity among bovine leukemia virus genotypes and its relation to humoral responses in hosts. AB - The existence of bovine leukemia virus (BLV) genotypes was investigated by restriction fragment length polymorphism (RFLP) analysis using bovine peripheral blood leukocytes collected from different geographical areas of Japan. For this purpose a nested polymerase chain reaction (PCR) for a 444 bp fragment of the envelope (env) gene was used because it was previously reported that this region might be responsible for the serological status in the host. The PCR products from 60 samples of BLV-infected cells were digested with endonucleases BamH I, Bgl I, Bcl I, Hae III and Pvu II. RFLP analysis demonstrated that there were six different genotypes of BLV present among cattle in Japan. In some herds PCR positive animals were infected with only one genotype, but in other herds a few genotypes were found. One genotype was dominant throughout infected cattle and it was also detected in neoplastic cells from three of four animals with lymphosarcoma and three cell lines persistently infected with BLV. Production of antibodies to BLV in each cattle was surveyed by agar gel immunodiffusion and indirect hemagglutination tests, and the results were compared with those obtained from PCR. No genotype related to decreased immunoreactivity was detected. The difference in anti-viral immune responses of each animal appears to be related to the infection stage and other host factors, not to genetic heterogeneity of the envelope gene. PMID- 12076832 TI - Vesicular stomatitis virus induces apoptosis at early stages in the viral cycle and does not depend on virus replication. AB - We detected apoptosis induction in the vesicular stomatitis virus (VSV) infected mammalian cell lines Vero-76, Cos-7, and BHK-21. Cell lines were analyzed by chromosomal DNA fragmentation and nuclear morphology. In order to determine the step in the viral cycle at which apoptosis of infected cells is triggered, chemical and physical agents were used to block viral infection at different times and then the apoptotic response of infected cells was examined. The treatment of Vero-76 infected cells with a lysosomotrophic agent, such as NH4Cl, was shown to abrogate virus apoptosis induction. On the other hand, VSV-induced apoptosis was not blocked by the presence of cycloheximide, suggesting that the de novo viral protein synthesis is not required for this process. UV-inactivated viruses were also capable of inducing apoptosis in Vero-76 cells, indicating that the activation of a programmed cell death process by VSV does not require viral replication. We conclude from these findings that VSV induces apoptosis at early stages of infection. PMID- 12076835 TI - Rescue of rubella virus replication-defective mutants using vaccinia virus recombinant expressing rubella virus nonstructural proteins. AB - The genome of rubella virus (RV) is translated into a polyprotein precusor, p200, of the nonstructural proteins (NSPs). This is proteolytically processed by a viral-encoded protease into two mature products, p150 and p90. p150 contains sequence corresponding to the predicted methyltransferase and protease activities, while p90 has sequence for the proposed helicase and RNA-dependent RNA polymerase activities. Processing of p200 is essential for RV viral replication. RV NSPs are responsible for viral RNA replication, in which a full length negative-strand RNA serves as the intermediate for the replication of positive-strand genomic RNA and the transcription of subgenomic RNA. Previously we demonstrated that p200 synthesizes negative- but not positive-strand RNA, and that cleavage products p150/p90 are required for efficient production of positive strand RNA. To determine whether p150 or p90 alone or together is involved in positive-strand RNA synthesis, vaccinia virus recombinants expressing individual NSPs were constructed and characterized. These were used in in vivo rescue experiments to complement replication-defective mutants in virus replication. A protease-inactive mutant was rescued by p200 or p150 provided in trans by using vaccinia virus recombinants. Thus this protease can function in trans. Rescue of cleavage-defective mutant by either p200 alone, or p150 plus p90 but not by p150 or p90 alone suggests that p150 and p90 function together as a replication complex in positive-strand RNA synthesis. PMID- 12076836 TI - Alterations and diversity in the cytoplasmic tail of the fusion protein of subacute sclerosing panencephalitis virus strains isolated in Osaka, Japan. AB - We determined the nucleotide sequence of the fusion (F) gene of three strains (Osaka-1, -2, and -3) of nonproductive variants of measles virus (MV). These viral strains were isolated in Osaka, Japan, from brain tissues of patients with subacute sclerosing panencephalitis (SSPE). Phylogenetic analysis revealed a close relationship among the three strains of SSPE virus. The cytoplasmic tail of the F protein, predicted from sequence analysis of the gene, is altered in all three SSPE strains when compared to the MV field strains. However, the extent and mode of alteration are different in each strain. The F protein of the Osaka-1 strain has six nonconservative amino acid substitutions and a 29-residue elongation of its cytoplasmic tail. The F protein of the Osaka-3 strain has two nonconservative substitutions and a 5-residue truncation of its C-terminus. Although the termination codon is not altered in the F protein of the Osaka-2 strain, five or six amino acids are changed in the cytoplasmic tail of the F protein of the two sibling viruses of this strain. The significance of the altered cytoplasmic domain of the SSPE viruses in the SSPE pathogenesis is discussed. PMID- 12076837 TI - Expression of Cardamom mosaic virus coat protein in Escherichia coli and its assembly into filamentous aggregates. AB - Cardamom mosaic virus (CdMV), a member of the genus Macluravirus of Potyviridae, causes a mosaic disease in cardamom. A polyclonal antiserum was raised against the purified virus and IgG was prepared. Electron microscopic studies on the purified virus showed flexuous filamentous particles of approximately 800 nm in length, typical of members of Potyviridae. The coat protein (CP) encoding sequence of the virus was expressed in Escherichia coli and the protein purified by affinity chromatography under denaturing conditions. The viral nature of the expressed CP was confirmed by positive reaction with anti CdMV IgG in a Western blot. The expressed CP aggregated irreversibly upon renaturation at concentrations above 0.07 mg/ml. The expression of the CP led to the formation of filamentous aggregates in E. coli as observed by immuno-gold electron microscopy. The filamentous aggregates were of 100-150 nm in length. Immuno-capture RT-PCR confirmed the absence of coat protein mRNA in the filamentous aggregates. Deletion mutations, which were expected to inhibit virus assembly, were introduced in the core region of the coat protein. However, these mutations did not improve the solubility of the CP in non-denaturing buffers. PMID- 12076839 TI - Castration in female rats modifies the development of the pilocarpine model of epilepsy. AB - Previous studies have shown that the susceptibility to pilocarpine-induced status epilepticus (SE) in female rats changes according to estrous cycle phases. These studies have also shown that following pilocarpine administration changes occur in gonadal, hypophyseal and hypothalamic hormones that could contribute for the sequence of the epileptic events. Accordingly, the present work aimed to investigate the role of sexual hormones withdrawal on the development of the pilocarpine model of epilepsy in female rats. With this purpose, castrated and non-castrated adult female Wistar rats were injected with pilocarpine and some characteristic parameters of the experimental model were observed. The results showed increased mortality after pilocarpine injection in the castrated rats when compared with non-castrated females. The latency period for SE onset and for the first spontaneous seizure was decreased in castrated when compared with non castrated animals. The mossy fiber sprouting measured by neo-Timm scale during the chronic period, reached grade 3 for castrated epileptic rats while the non castrated epileptic rats showed grade 2. Our results indicate that castration interferes with the epileptogenesis in the pilocarpine model of epilepsy suggesting that female sexual hormones could have protective effects against pilocarpine-induced SE. PMID- 12076840 TI - Clobazam shows a different antiepileptic action profile from clonazepam and zonisamide in Ihara epileptic rats. AB - PURPOSE: Clobazam (CLB, 1,5-benzodiazepine, 1,5-BZP) has been reported to show unique antiepileptic action profile distinguishing from standard 1,4-BZPs. To further elucidate the action profile of CLB, its effects on the abnormal circling fits (ACFs) and generalized tonic-clonic convulsions (GTCs) in Ihara epileptic rats (IERs), a genetically epileptic mutant, were examined in comparison with conventional antiepileptic drugs (AEDs), a 1,4-BZP, clonazepam (CZP) and a non BZP, zonisamide (ZNS). METHODS: The incidence of ACFs or GTCs in IERs was recorded automatically by the computer-assisted behavior monitoring system (COBAS N-IV) before, during and after the drug treatment period for 5 days in each. The drugs were orally administered twice daily. The daily and total incidences of ACFs or GTCs were calculated every each period in each dose group. The incidences of various behaviors such as feeding, gnawing and scratching recorded simultaneously were used for evaluating the behavioral activity (BA). RESULTS: CLB (30 and 60 mg/kg) prevented the appearance of ACFs and GTCs without affecting BAs. CZP (1 and 3 mg/kg) suppressed the occurrence of ACFs but induced no effect on the incidence of GTCs. Furthermore, it inhibited BAs at the same doses. ZNS (15 mg/kg) suppressed GTCs but little ACFs without affecting BA. CONCLUSION: CLB exhibited a different action profile from CZP and ZNS in a novel epileptic mutant, IERs, and was expected to be a useful AED superior to 1,4-BZPs in clinical practice. PMID- 12076841 TI - Gabapentin and vigabatrin increase GABA in the human neocortical slice. AB - The effects of antiepileptic drugs, gabapentin and vigabatrin, on gamma aminobutyric acid (GABA) concentrations were studied in human (n=14) and rat (n=6) neocortical slice preparations. In this study, neocortical slices were incubated with gabapentin, vigabatrin or no drugs for 3 h in an oxygenated environment. Proton magnetic resonance spectroscopy (MRS) of perchloric acid (PCA) extracts was used to measure GABA concentrations. Vigabatrin increased cellular GABA concentrations in both human and rat neocortical slices by 62% (P<0.001) and 88% (P<0.03), respectively. Gabapentin significantly increased GABA concentrations by 13% (P<0.02) in human neocortical slices made from tissue resected during epilepsy surgery. However, in the rat neocortical slice exposed to the same conditions as the human tissue, gabapentin did not increase GABA significantly. These results confirm our MRS studies in vivo that gabapentin increases GABA levels in epileptic patients, but has minimal or no effect in a healthy rodent model. Caution must be used in extrapolating negative results obtained in rodent models to the human condition. PMID- 12076842 TI - Time course of lamotrigine de-induction: impact of step-wise withdrawal of carbamazepine or phenytoin. AB - OBJECTIVE: The objective of the present analysis is to examine lamotrigine (LTG) pharmacokinetics both during polytherapy with enzyme inducing antiepileptic drugs and to evaluate the time course of de-induction following the step-wise withdrawal of enzyme inducers. BACKGROUND: LTG pharmacokinetics can be significantly influenced by concomitant AEDs, and the addition of enzyme inducers can markedly increase LTG clearance, thereby reducing serum concentrations. A clinically relevant question is how will LTG clearance and resulting plasma concentrations be altered during concomitant enzyme inducer withdrawal/conversion process. DESIGN/METHOD: As part of a previously published, active-control, LTG monotherapy trial, dose and plasma concentration data for LTG, carbamazepine (CBZ) or phenytoin (PHT) were obtained. Following the attainment of a LTG target dose of 500 mg/day, CZB or PHT were withdrawn in weekly 20% decrements. Following inducer withdrawal, LTG was then continued as monotherapy for an additional 12 weeks. Plasma concentrations and daily doses of LTG, CBZ, or PHT were obtained at regularly scheduled study visits during inducer withdrawal and during LTG monotherapy. Pharmacokinetic analysis of the plasma concentration data was done to determine the time-course and effect of inducer plasma concentration on LTG oral clearance (Cl(o)), where LTG Cl(o) was estimated as the dose/concentration ratio. RESULTS: Of the 156 patients enrolled in this trial, 76 were assigned to LTG arm, 43 completed the withdrawal to monotherapy phase with 28 successfully completing the study. In a subset analysis of completers, LTG Cl(o) determined prior to withdrawal of the inducers was significantly greater in patients (n=28) on LTG+PHT (160% increase) than in those (n=48) receiving LTG+CBZ (62% increase): 1.77+/-0.77 vs. 1.06+/-0.41 ml/min/kg, respectively, p=0.017. The significant reduction in LTG Cl(o) occurred only when CBZ plasma concentrations reached approximately 2 microg/ml or PHT plasma concentrations reached zero. CONCLUSIONS: Mean LTG plasma concentrations will approximately double following the withdrawal PHT; however increases of only 60% may occur following the withdrawal of CBZ. Importantly, these data suggest that LTG concentrations would not be expected to increase until the concomitant inducer is almost completely removed. PMID- 12076843 TI - Aminophylline exacerbates status epilepticus-induced neuronal damages in immature rats: a morphological, motor and behavioral study. AB - Adenosine is an endogenous modulator that has an inhibitory effect on neuronal activity. The aim of this work was to investigate the role of aminophylline, an adenosine receptor antagonist, on the long-term effects of status epilepticus (SE) in the developing brain. Four groups of rats at the postnatal age of 12 days were intraperitoneally administered with saline, aminophylline (50 mg/kg), lithium-pilocarpine (Li-PC) (3 mEq/kg-60 mg/kg), and Li-PC plus aminophylline, respectively. The four groups were tested for spatial memory using the Morris water maze task at P80 and motor performance by the Rotarod test at P100. The brains were then analyzed with cresyl violet stain for histological lesions and evaluated for mossy fiber sprouting with the Timm stain. At the acute stage, all rats subjected to Li-PC developed SE and no seizures were elicited in the saline treated or aminophylline-treated rats. The seizure duration was longer in the Li PC plus aminophylline group (346.9+/-32.7 min) as compared with that in the Li-PC group (265.2+/-9.8 min). The difference of mortality was not significant. Rats without seizures exhibited no motor imbalance, spatial deficits, or morphological changes. The rats with Li-PC-induced SE demonstrated spatial memory deficits without motor incoordination or morphological changes. However, the rats subjected to Li-PC plus aminophylline exhibited motor impairment and morphological changes, including neuronal cell loss in CA1 area and increased mossy fiber sprouting in CA3 area. In addition, the rats of Li-PC plus aminophylline had greater spatial memory deficits than that seen in rats with Li PC. We concluded that an adenosine receptor antagonist, such as aminophylline, had synergistic effects on the SE-induced long-term deficit of cognition and motor performance in the developing brain. The present study may provide experimental evidence and lead to novel therapeutic interventions. PMID- 12076844 TI - Hydroethidine detection of superoxide production during the lithium-pilocarpine model of status epilepticus. AB - Hydroethidine is reported to be selectively oxidized to ethidium by superoxide. Using digital imaging and fluorescence microscopy it is possible to evaluate neuronal ethidium accumulation in specific brain regions of rats damaged in the lithium-pilocarpine model of status epilepticus. Intravenous or intraperitoneal administration of hydroethidine prior to 1 h of status epilepticus produced diffuse cytosolic distribution of ethidium fluorescence suggesting an increased neuronal production of superoxide that was not observed in control animals. A significantly increased number of neurons with the enhanced ethidium fluorescence was observed in parietal cortex, piriform cortex, perirhinal cortex, lateral amygdala, mediodorsal thalamus and laterodorsal thalamus, suggesting superoxide as a mechanism of neuronal injury in those regions. Other regions injured by lithium-pilocarpine seizures, such as the basolateral amygdala and hippocampus, did not demonstrate the enhanced neuronal ethidium fluorescence. In such regions it is possible that superoxide is not a mechanism of injury or that 1 h of status epilepticus is not sufficient to produce superoxide or other reactive oxygen species. PMID- 12076846 TI - Influence of cytochrome P450 induction on the pharmacokinetics and pharmacodynamics of remacemide hydrochloride. AB - Remacemide hydrochloride (RMD) is a putative anticonvulsant agent with an active metabolite, desglycinyl-remacemide (DGR) and a broad spectrum of activity in experimental seizure models. In clinical trials, however, the efficacy of RMD is questionable. In the case of add-on studies, the inconclusive findings may be related to pharmacokinetic interactions between RMD and established antiepileptic drugs. We have investigated the influence of cytochrome P450 (CYP(450)) induction following repeated treatment with phenobarbital (PB) on the pharmacokinetics and pharmacodynamics of RMD in mice. Pre-treatment with PB (80 mg/kg; once daily for 4 days) significantly increased CYP(450) content and activity in mouse liver. This was associated with a consistent reduction in the brain concentrations of both RMD and DGR and attenuation of the anticonvulsant effects of RMD in the maximal electroshock model. Pharmacokinetic analysis suggested that DGR was proportionately more susceptible to CYP(450) induction than the parent compound. As the principal active moiety, the selectively enhanced metabolism of DGR under induced conditions may underlie the debatable findings of add-on trials with RMD in refractory epilepsy. However, this hypothesis does not explain the similarly questionable efficacy of RMD monotherapy in newly diagnosed epilepsy, an observation that may have wider pharmacological implications. PMID- 12076845 TI - Effects of acute inhibition of fatty acid oxidation on latency to seizure and concentrations of beta hydroxybutyrate in plasma of rats maintained on calorie restriction and/or the ketogenic diet. AB - The present study was designed to evaluate the effects of acute inhibition of fatty acid oxidation on plasma levels of beta hydroxybutyrate and latency to PTZ induced seizures in ad libitum- (AL), calorie-restricted normal rodent chow- (CR), and calorie-restricted ketogenic diet (KD)-fed young rats. Young (day 23) Sprague-Dawley rats were fasted for 8 h and then fed their respective diets for 21 days. On day 21 of the diet rats in each group received either saline or the fatty acid oxidation inhibitor mercaptoacetate (MA; 46 mg/kg intraperitoneally (i.p.). Two hours later, all rats received pentylenetetrazole (PTZ; 10 mg/kg; i.p.) every 10 min until seizure onset. Results demonstrated that KD-fed rats had the longest (P<0.05) latency to PTZ-induced seizures. KD-fed rats administered an acute dose of MA had lower (P<0.01) levels of beta hydroxybutyrate in plasma and shorter latency to PTZ-induced seizures compared with control KD-fed rats. However, there was not a significant positive correlation (P>0.10) between plasma beta hydroxybutyrate and latency to seizure, suggesting that beta hydroxybutyrate may be indirectly involved in the antiseizure effects of the KD. Fatty acid oxidation inhibition represents an experimental manipulation that may allow for more precise establishment and evaluation of levels of beta hydroxybutyrate in plasma necessary for antiseizure effects of the KD. PMID- 12076847 TI - Evidence against association between arachnoid cysts and epilepsy. AB - Arachnoid cysts are often discovered incidentally in MRs performed for a variety of reasons. In this study, we have attempted to determine a possible relationship between the arachnoid cyst, seizure type and EEG abnormalities. Eight patients were classified according to the Classification of Epilepsies and Epileptic Syndromes of ILAE as idiopathic generalized or localized epilepsy: three as idiopathic generalized epilepsy, three as Rolandic epilepsy and two as juvenile myoclonic epilepsy. Two patients with rare nocturnal seizures had normal EEGs in the awaking and sleep stage. The EEGs of five patients with simple or complex partial seizures revealed focal epileptiform abnormalities, but only one patient had the same location with the arachnoid cyst. Five patients with symptomatic epilepsy had diffuse slowing in their EEGs and only one patient had seizure focus in the EEG that was contralateral to the arachnoid cyst. According to our study, seizure type and EEG abnormality corresponds to arachnoid cyst location in only one patient. Therefore, we suggest that arachnoid cysts may not be related to a specific seizure type and EEG focus. PMID- 12076849 TI - Polyspike discharges in Lennox-Gastaut syndrome. PMID- 12076851 TI - [Content validity of an information system used for head injury rehabilitation based on an analysis of medical records]. AB - OBJECTIVE: This study examines the content validity of the TCC-Quebec Information System based on an analysis of rehabilitation medical records. The goal was to determine the agreement between the variables of the TCC-Quebec Information System identified by experts and based on an extensive literature review and those found in medical records. METHOD: The medical records of 82 adults with a head injury were reviewed. The individuals had been hospitalized from 1997 to 1998 at three different acute care facilities or three rehabilitation centers. The abstractor determined if the information pertaining to a variable (e.g. personal history, impairments, or disabilities relating to sensori-motor function) was present in the record. A standardized and reliable procedure was used to ensure the quality of data extraction. The percentage of variables found in the medical records and the number of records in which each variable was documented were calculated for each clinical setting (acute care or rehabilitation) and for the different geographical regions. RESULTS: The results suggest that a large discrepancy exists between what experts desired to be included in the information system and what is really documented clinically. No discrepancy exists between the different regions. Only 23% of variables were found in more than 70% of records. CONCLUSION: This study provides recommendations about the most relevant variables to be included in an information system based on clinicians'information needs and the clinical reality. As such, these results should facilitate the use and implementation of the information system under study. PMID- 12076852 TI - [Feasability of a back school assessment programme]. AB - OBJECTIVES: Testing the feasability of a back school assessment programme in two populations of people suffering with chronic low back pain. MATERIAL: Twenty-nine patients were randomly included in a chronic low back pain assessment programme (15 patients waiting for back school and 14 patients after back school). METHODS: The programme was made of the measure of the age, the sex, the body mass index, the pain (VAS, St Antoine Hospitals Pain Questionnaire - SAPQ), the anxiety, the depression, the RIII nociceptive reflex, the fingertip-floor distance, the strength of the flexors and of the extensors of the lumbar spine (CybexR 6000 isocinetic dynamometer), the lumbar function (EIFEL, Dougados), the Dallas self questionnaire. The Back School Education programme was made of five sessions (information, ergonomics, extension exercises). RESULTS: Comparing the two populations we did not observe significant differences concerning the age, the body mass index, the anxiety and depression levels, the pain (VAS, SAPA, RIII nociceptive reflex), the lumbar stiffness, the lumbar disability, the quality of life; the patients who had achieved back school had a peak torque of the flexors and a ratio flexors/extensors significantly lower. Comparing men and women we observed significant differences in the SAPQ and the muscles strength. The SAPQ was correlated with the depression and anxiety levels, the lumbar disability, the peak torque of the flexors. The VAS was correlated with the age, the lumbar stiffness, the depression level, the peak torque of the flexors. The RIII nociceptive reflex was correlated with the ratio flexors/extensors. CONCLUSION: This study present some biases but this objective assessment of chronic low back pain appeared as feasible. The pain must be investigated in term of intensity, expression, alleviation. The impairment of the flexors muscles in women and after back school has to be confirmed. The different tests are relevant to determinate the efficiency of the back school programmes. PMID- 12076853 TI - [Effectiveness and compliance to a center-based short rehabilitation program with a home-based program for chronic low back pain]. AB - OBJECTIVES: To assess the effectiveness and compliance to a short center-based intensive rehabilitation program followed by a home-based program for chronic low back pain. METHODS: Open prospective study in patients with chronic low back pain. The program included five weekly meetings of 12 day. Followed by a home based program consigned on a booklet. Evaluation of deficiencies (lumbar and radicular pains on an analog visual scale, Schober Mac Rae test, distance finger ground, Shirado test, Sorensen test), disabilities (Quebec functional index), and handicap (analog visual scale) were assessed at J0, five weeks, three months and one year. Compliance was assessed quantitatively (a number of exercises, duration of the meetings, numbers meetings per week) and qualitatively (semi-quantitative ordinal scale). Patient's global opinion on the effectiveness of the program was collected on a semi-quantitative ordinal scale. RESULTS: 60 patients (20 women), aged of 45 +/- 10,4 years, were included. Duration of the symptoms was 110 +/- 123 months. At five weeks, a significant improvement of all parameters was observed. This improvement was maintained at three months. At 12 months, there was no more significant improvement. Compliance to the program was 64,4%. 40.7% of the patient did not realize their exercises correctly. The majority gave up back muscle isometric reinforcement exercises. Score of Shirado test, Quebec index and EVA handicap were significantly different in the group realizing the home-base program. At one year, 79% of the patients regarded this program as useful. CONCLUSION: Although quantitative compliance with the home-based program can be considered as satisfactory, the effectiveness of this intensive rehabilitation program of lumbar decrease after three months. This can be explained by a fast deterioration of qualitative compliance. The effectiveness of a short supervised program between 3 and 6 months associated a more powerful message of motivation must be evaluated. PMID- 12076855 TI - Feinstone SM, Kapikian AZ, Purcell RH. Hepatitis A: detection by immune electron microscopy of a virus like antigen associated with acute illness [Science 1973;182:1026-1028]. PMID- 12076856 TI - Proliferation of rat small hepatocytes after long-term cryopreservation. AB - BACKGROUND/AIMS: The demand for clinical use of hepatocytes is escalating because cell transplantation will be an alternative to orthotopic liver transplantation and the shortage of liver donors is a serious problem throughout the world. However, the supply of fresh differentiated hepatocytes is limited and methods for cryopreservation of hepatocytes that can proliferate with hepatic functions are not satisfactorily established. METHODS: Colonies of small hepatocytes were collected and then maintained at -80 degrees C for more than 6 months. Albumin secretion and mRNA expression of thawed cells were measured by enzyme linked immunosorbent assay and Northern blotting, respectively, and the expression of hepatic functions was examined by immunoblotting. The ultrastructure of cryopreserved cells was also examined. RESULTS: About 60% of the cryopreserved colonies attached on dishes and then proliferated. The average area of small hepatocyte colonies was about 7.5 times larger at day 15 than at day 1. Albumin production increased with time in culture. In addition, the cells produced other serum proteins such as transferrin and fibrinogen, and expressed carbamoyl phosphate synthetase I and tryptophan 2,3-dioxygenase. CONCLUSIONS: Small hepatocytes maintain growth ability and hepatic differentiated functions even after long-term cryopreservation. PMID- 12076857 TI - Effects of long-term oral misoprostol administration on hepatic amino acid nitrogen metabolism in patients with cirrhosis. AB - BACKGROUND: The acute infusion of a Prostaglandin of E series 1 (PGE1) analogue results in nitrogen sparing in cirrhosis. AIMS: To test the effects of long-term oral PGE1 on hepatic and whole-body nitrogen metabolism. PATIENTS AND METHODS: Ten patients with advanced cirrhosis were studied in paired experiments, before and 30-50 days after oral misoprostol therapy. alpha-Amino-nitrogen levels and urea-nitrogen synthesis rate were measured in the post-absorptive state and in response to continuous alanine infusion (2 mmol/kg per hour for 4.5h). Data were used to compute the functional hepatic nitrogen clearance, i.e. the slope of the regression of alpha-amino-N levels to urea-N synthesis rate, and the apparent nitrogen exchange. RESULTS: Misoprostol reduced urea-N synthesis rate (during fasting and in response to alanine), resulting in a positive nitrogen exchange. The functional hepatic nitrogen clearance slightly increased, and the regression line was rightwards shifted, indicating a reduced urea synthesis rate at any alpha-amino-N concentration. Amino acid- and ammonia-N did not accumulate in plasma. No systematic effects on insulin and glucagon were observed. CONCLUSIONS: Data are consistent with a nitrogen sparing mechanism of misoprostol, not mediated by hormone levels. These effects may be beneficial in clinical hepatology, and need to be tested in controlled trials. PMID- 12076858 TI - Inhibition of renin-angiotensin system attenuates liver enzyme-altered preneoplastic lesions and fibrosis development in rats. AB - BACKGROUND/AIMS: It is suggested that the renin-angiotensin system (RAS) is involved in tumor development and fibrogenesis. The aim of the present study was to examine the effect of RAS inhibition on the liver enzyme-altered preneoplastic lesions and fibrosis development. METHODS: The effects of the clinically used angiotensin-I converting enzyme inhibitor (ACE-I), perindopril (PE), on two different rat model of liver carcinogenesis models induced separately by diethylnitrosamine (DEN) and a choline-deficient L-amino acid-defined (CDAA) diet were studied. This CDAA model was also used to elucidate the effect of PE on liver fibrosis development. RESULTS: The immunohistochemical evaluation revealed that the glutathione S-transferase placental form (GST-P), and gamma glutamyltransferase (GGT)-positive preneoplastic foci significantly decreased in the livers of the PE-treated groups. In CDAA-induced liver fibrosis model, PE revealed a marked inhibitory effect of liver fibrosis development. The hepatic hydroxyproline, serum fibrosis markers, alpha-smooth muscle actin (alpha-SMA) immunopositive cells in number, and alpha-(III) pro-collagen mRNA expression were significantly suppressed by PE treatment. These inhibitory effects of PE were achieved even at a clinically comparable dose (2 mg/kg per day). CONCLUSIONS: These results suggested that the RAS is involved in liver carcinogenesis and fibrosis development. PMID- 12076859 TI - Effects of amrinone on hepatic ischemia-reperfusion injury in rats. AB - BACKGROUND/AIMS: The present study was designed to investigate the effect of amrinone, a phosphodiesterase III inhibitor, on hepatic ischemia-reperfusion injury in rats. METHODS: Amrinone was infused at a rate of 20 or 100 microg/kg/min, and 60-min partial ischemia was induced. The effects of amrinone on hemodynamic status, hepatic tissue cyclic adenosine 5'-monophosphate (cAMP), hepatic tissue blood flow, platelet aggregation and plasma levels of transaminase were examined. The expression of intercellular adhesion molecule-1 (ICAM-1) and myeloperoxidase activity were analyzed and histological examination was performed in the injured liver. The cumulative survival rates for 14 days were also examined. RESULTS: Hemodynamic status was not affected by amrinone. The levels of cAMP during reperfusion were significantly higher in rats with amrinone. Hepatic tissue blood flow during reperfusion was increased and platelet aggregation was inhibited by amrinone. The expression of ICAM-1 mRNA and protein in the injured liver was suppressed in rats with amrinone. The levels of transaminase, necrotic changes and myeloperoxidase activity were suppressed after reperfusion and higher survival was achieved in the rats treated with amrinone. CONCLUSIONS: Amrinone protected against ischemia-reperfusion injury of the liver in the present model. PMID- 12076860 TI - Up-regulated expression of fractalkine and its receptor CX3CR1 during liver injury in humans. AB - BACKGROUND/AIMS: Little is known about the role of fractalkine (CX3CL1) in the liver. The aim of this study was to investigate the expression patterns of fractalkine and its receptor CX3CR1 in normal human liver and in conditions of injury. METHODS: Distribution and expression of fractalkine and its receptor were investigated using immunohistochemistry, in situ hybridization, flow cytometry and reverse transcriptase-polymerase chain reaction. In vitro experiments were conducted in HepG2 cells. RESULTS: Both fractalkine and CX3CR1 were up-regulated during chronic injury, in areas of portal and lobular inflammation. In severe acute hepatitis, fractalkine and CX3CR1 were expressed at high levels not only in areas of inflammation but also in regenerating epithelial cells within bile duct like structures, which showed co-expression of fractalkine and cytokeratin-7 or CX3CR1. The human hepatocarcinoma cell line HepG2 expressed fractalkine at the gene and protein level, and HepG2-conditioned medium was chemotactic for cells overexpressing CX3CR1. Transcripts for CX3CR1 were detected in HepG2, and exposure of these cells to recombinant fractalkine induced cell migration. CONCLUSIONS: This study shows that the fractalkine system is up-regulated during liver damage, and suggests that fractalkine may play a role in the recruitment and adhesion of inflammatory cells and in the biology of liver epithelial cells. PMID- 12076861 TI - Gene deletion of NF-kappa B p50 does not alter the hepatic inflammatory response to ischemia/reperfusion. AB - BACKGROUND/AIMS: Nuclear factor kappa B (NF-kappa B) is a primary regulator of gene expression and is activated during hepatic ischemia/reperfusion injury. The objective of the present study was to determine whether activation of NF-kappa B is causally related to the induction of the acute inflammatory response induced by hepatic ischemia/reperfusion. METHODS: Wild-type (p50(+/+)) and NF-kappa B p50 deficient (p50(-/-)) mice underwent hepatic ischemia/reperfusion. NF-kappa B activation was determined by electrophoretic mobility shift assay. Hepatic neutrophil accumulation was measured by liver myeloperoxidase content. Hepatocellular injury was assessed by serum level of alanine aminotransferase and liver histology. RESULTS: In p50(+/+) mice, ischemia/reperfusion induced marked activation of NF-kappa B consisting of p50/p65 heterodimers. In contrast, NF kappa B activation in livers from p50(-/-) mice was abrogated, but p65 was observed in nuclear extracts. Despite amelioration of NF-kappa B activation there was no significant difference between p50(+/+) and p50(-/-) mice in expression of TNF alpha and MIP-2, liver accumulation of neutrophils or hepatocellular injury. CONCLUSIONS: Gene deletion of NF-kappa B p50 does not alter the hepatic inflammatory response to ischemia/reperfusion. Despite abrogation of DNA-binding by the NF-kappa B p50/p65 complex, p65 was still observed in nuclear extracts suggesting that there may be functional redundancy amongst members of the Rel protein family in order to preserve the inflammatory response. PMID- 12076862 TI - In situ detection of lipid peroxidation and oxidative DNA damage in non-alcoholic fatty liver diseases. AB - BACKGROUND/AIMS: Although oxidative stress is an important candidate in the pathogenesis of non-alcoholic fatty liver disease (NAFLD), the localization and pathological significance of oxidative stress-induced cellular damage in NAFLD remains unclear. METHODS: Hepatic expression of 4-hydroxy-2'-nonenal (HNE) and 8 hydroxydeoxyguanosine (8-OHdG), as reliable markers of lipid peroxidation and oxidative DNA damage, respectively, was immunohistochemically investigated in NAFLD and the results were compared with histological findings. RESULTS: While no HNE adducts were observed in control livers, they were frequently detected in NAFLD. In NASH, the localization of the adducts was in the cytoplasm of sinusoidal cells and hepatocytes with a predominance in zone 3. The grade of necro-inflammation as well as the stage of fibrosis significantly correlated with the HNE index. Regarding 8-OHdG, although no 8-OHdG expression was observed in normal liver and only a few in fatty liver, 11 of 17 cases (64.7%) with NASH exhibited nuclear expression of 8-OHdG in hepatocytes and sinusoidal cells in areas of active inflammation. The 8-OHdG index significantly correlated with the grade of necro-inflammation. CONCLUSIONS: Oxidative cellular damage occurs frequently in livers with NAFLD and may be associated with some clinico pathological features of NAFLD including liver fibrosis and possibly, hepatocarcinogenesis. PMID- 12076863 TI - Activation of NF-kappa B, AP-1 and STAT transcription factors is a frequent and early event in human hepatocellular carcinomas. AB - BACKGROUND/AIMS: Hepatitis B and C viruses, two inducers of hepatocarcinomas, have been shown to activate AP-1, NF-kappa B and STAT in vitro, but no detailed information on the activity of these transcription factors in vivo have been provided. METHODS: We have measured the DNA binding activity of these transcription factors in the peri-tumoral and the tumoral parts of 15 primary liver cancers, of viral or non-viral etiologies, and in five hepatic metastases using electrophoretic mobility shift assays. RESULTS: AP-1, NF-kappa B and STAT binding activities were increased in the peritumoral tissue, compared with histologically normal livers in 73, 87 and 70%, respectively, of the cases. A further activation of AP-1, NF-kappa B, but not STAT binding in the tumoral parts was detected in 40 and 80%, respectively, of the cases. A close correlation was found between JunD and c-Jun levels and AP-1 binding activity at the tumoral stage. By contrast, AP-1 and NF-kappa B binding activities were low or only slightly elevated in the peri-tumoral and the tumoral tissue of metastases. CONCLUSIONS: Early activation of AP-1, NF-kappa B and STAT contributes probably to the acquisition of a transformed phenotype during hepatocarcinogenesis, whatever the etiology. PMID- 12076864 TI - Improved functional reserve of hypertrophied contra lateral liver after portal vein ligation in rats. AB - BACKGROUND/AIMS: We assessed the functional capacity of hypertrophied liver after portal vein ligation (PL) in a test group of rats compared to a control group (without PL) having the same size liver. METHODS: The portal veins of the left and median lobes in the test group were ligated in an initial operation. Four days after the PL, the liver volume of the posterior caudate lobe (5%) increased two-fold, accounting for 10% of the liver. Then a 90% hepatectomy was performed, leaving only the hypertrophied posterior caudate lobe. Rats in a sham group underwent a 90% hepatectomy 4 days after having laparotomy, leaving the normal anterior and posterior caudate lobes (10%). RESULTS: The survival rate for the PL group was significantly higher than for the sham group at 4 days after hepatectomy (56.3 and 26.7%, P<0.05). The regeneration ratio and the proliferating cell nuclear antigen (PCNA) labeling index in the PL group was markedly higher than in the sham group 24h after hepatectomy. CONCLUSIONS: Hypertrophied liver at 4 days after PL still showed liver regeneration. Regenerating liver provided greater tolerance for extended hepatectomy than normal liver. This is because of the induced rapid regeneration of the remaining liver after hepatectomy. PMID- 12076865 TI - Improved prognosis of postoperative hepatocellular carcinoma patients when treated with functional foods: a prospective cohort study. AB - BACKGROUND/AIMS: Active hexose correlated compound (AHCC) is a newly developed functional food. In vitro experiments have shown that AHCC enhances natural killer cell activity, and may be considered a potent biological response modifier in the treatment of cancer patients. However, the effects of AHCC in a clinical setting have not been reported. We seek to determine whether AHCC can improve the prognosis of hepatocellular carcinoma (HCC) patients following surgical treatment. METHODS: A prospective cohort study was performed from February 1, 1992 to December 31, 2001. A total of 269 consecutive patients with histologically confirmed HCC were studied. All of the patients underwent resection of a liver tumor. Time to treatment failure (disease recurrence or death) and ten parameters related to liver function after surgery were examined. RESULTS: Of the 269 patients, 113 received AHCC orally after undergoing curative surgery (AHCC group). The AHCC group had a significantly longer no recurrence period (hazard ratio (HR), 0.639; 95% confidence interval (CI), 0.429-0.952; P=0.0277) and an increased overall survival rate (HR, 0.421; 95% CI, 0.253-0.701; P=0.0009) when compared to the control group by Cox's multivariate analysis. CONCLUSIONS: This study suggests that AHCC intake can improve the prognosis of postoperative HCC patients. PMID- 12076866 TI - Heparin accelerates liver regeneration following portal branch ligation in normal and cirrhotic rats with increased plasma hepatocyte growth factor levels. AB - BACKGROUND/AIMS: Heparin is widely used as a general anticoagulant, and has been recently reported to elevate plasma hepatocyte growth factor (HGF) levels by releasing HGF sequestrated in the extracellular matrix. Therefore, we investigated the effects of heparin administration on liver regeneration following portal branch ligation (PBL) in normal and cirrhotic rats. METHODS: Dimethylnitrosamine-induced cirrhotic rats and control rats underwent portal ligation of the left lateral and median branches, followed by intraperitoneal heparin injections, every 12 h. To examine the feasibility of an extensive hepatectomy in the cirrhotic livers, cirrhotic rats with or without heparin treatment underwent resection of occluded lobes at 72 h after the PBL. RESULTS: Heparin injections significantly augmented liver regeneration after PBL in both normal and cirrhotic rats, following an increase in hepatocellular DNA synthesis at 24 h after the PBL. The plasma HGF concentrations were elevated by heparin treatment in both groups. In addition, heparin administration dramatically improved the survival rate after an extensive hepatectomy in the cirrhotic rats. CONCLUSIONS: Heparin treatment significantly accelerated liver regeneration following the PBL, with an increase in the plasma HGF levels in both normal and cirrhotic rats. Heparin administration may make an extensive hepatectomy clinically feasible even for cirrhotic livers. PMID- 12076867 TI - Lack of effect of intravenous pamidronate on fracture incidence and bone mineral density after orthotopic liver transplantation. AB - BACKGROUND/AIMS: Increased rates of bone loss and fracture have been reported after liver transplantation. The aim of this study was to investigate the effects of a pre-transplant infusion of pamidronate on fracture incidence and bone loss during the first year after transplantation. METHODS: Ninety-nine adults awaiting orthotopic liver transplantation (OLT) were randomised to pamidronate or no treatment. Spinal X-rays were obtained at baseline and after 12 months. Bone mineral density (BMD) was measured at the lumbar spine (L1-4) and femoral neck at baseline, and 3, 6, and 12 months after OLT. RESULTS: The incidence of fractures in the first year after OLT was 8%, four patients within the pamidronate treated group and two in the untreated group developing fractures (P=0.40). No significant spinal bone loss occurred in either group during the first year. However, significant and sustained bone loss occurred at the femoral neck in both groups. No significant differences were seen between pamidronate treated or untreated groups at either site. CONCLUSIONS: Pamidronate in the regimen used had no significant effect on fracture rate or BMD post-transplant. The low incidence of fracture and absence of spinal bone loss indicate that bone disease after liver transplantation may be less common than previously reported. PMID- 12076868 TI - Decrease in hepatic CD56(+) T cells and V alpha 24(+) natural killer T cells in chronic hepatitis C viral infection. AB - BACKGROUND/AIMS: The intrahepatic immune system is likely to play a key role in determining the outcome of hepatitis C virus (HCV) infection. The hepatic lymphocyte repertoire is characterised by high CD8/CD4 T cell ratios and large numbers of gamma delta T cells, natural killer (NK) cells, NK T cells and NK receptor-positive T cells. It is not known which of these populations contribute to immunity against HCV or immune pathology. METHODS: To explore the relative contributions of lymphocyte subpopulations, we have compared the intrahepatic lymphocyte repertoires and cytokine expression in 13 patients with mild chronic hepatitis C infection, 14 with end-stage hepatitis C cirrhosis and five histologically normal livers by flow cytometry and immunohistochemistry. RESULTS: CD4(+) T cells bearing alpha beta T cell receptors (TCR) were significantly expanded in livers with chronic HCV infection while CD56(+) alpha beta T cells and V alpha 24 TCR-positive T cells were significantly depleted. Expanded CD4(+)T cells were predominantly Th1 cells, producing interferon-gamma but not interleukin-4. CONCLUSIONS: Failure to resolve HCV infection may be due to deficient innate and/or memory immune responses, while Th1 cells may mediate immune pathology. PMID- 12076869 TI - High doses of interferon in combination with ribavirin are more effective than the standard regimen in patients with HCV genotype 1 chronic hepatitis. AB - BACKGROUND/AIMS: The aim of the present, open-labelled, controlled study was to determine whether 5 MU of interferon (IFN) alpha 2b combined with a standard dose of ribavirin might increase the rate of viral clearance in all patients with chronic HCV hepatitis or at least in those with an unfavourable genotype. METHODS: A total of 298 previously untreated patients with chronic hepatitis C were randomized to 5 or 3 MU of interferon alpha 2b 3 times per week with 1000 1200 mg of ribavirin daily (148 and 150 patients, respectively). Patients were treated for 12 months and observed for 6 months posttreatment. RESULTS: In patients infected with HCV genotype 1, the sustained virologic response was 37.8% (95% CI 27.3-48.1) with IFN 5 MU and 19.2% (95% CI 10.1-28.2) with IFN 3 MU (P=0.008). Out of 45 sustained responders with genotype 1, 31 (69%) had received 5 MU and 14 (31.1%) the standard 3 MU dose of IFN in combination with ribavirin (P=0.01). Of the 86 responders infected with genotype non-1, 39 (45.3%) were from the 5 MU IFN group and 47 (54.6%) were from the 3 MU IFN group; these figures were not significant. At the multivariate analysis of baseline features for all patients, the variables with an independent effect for a sustained response were genotype non-1 (odds ratio (OR) 3.98, 95% CI 2.36-6.40), and the histological grading (score 0-2) (OR 2.48, 95% CI 1.12-5.51) and staging (score 0-1) (OR 1.73, 95% CI 1.02-2.95). For patients with genotype 1 only the high regimen of IFN entered the model (OR 2.39, 95% CI 1.13-5.05), whereas for patients with genotype non-1 an age of <40 years (OR 2.64, 95% CI 1.23-5.70) and staging (score 0-1) (OR 2.38, 95% CI 1.07-5.28) were independent predictors of a sustained response. CONCLUSIONS: Our study suggests that when treating naive patients with genotype 1, there is a significant increase in the rate of sustained virologic clearance by increasing the dose of IFN given in combination with ribavirin. PMID- 12076870 TI - Histological and virological features and follow-up of hepatitis C virus carriers with normal aminotransferase levels: the Italian prospective study of the asymptomatic C carriers (ISACC). AB - BACKGROUND/AIMS: To evaluate demographic characteristics, liver histology and virological features of hepatitis C virus (HCV) carriers with normal alanine transaminase (ALT) levels. METHODS: A nationwide prospective study was started in 1997. Four Italian centres have participated in this study. RESULTS: Eight hundred and eighty subjects entered the study. One hundred and eighty-nine (21.5%) were excluded during the follow-up because of ALT increase. Among the 691 patients with persistent ALT normality, 72% were females. An overall prevalence of genotype 2 was found (52%). Normal liver was found in 17% of the patients; 34% had minimal chronic hepatitis, 44% mild hepatitis, 4% moderate to severe hepatitis, and 1% had cirrhosis. Clinical and virological features did not differ between subjects with ALT flares and those with persistently normal ALT. Baseline ALT levels have no effects on liver histology and clinical outcome. CONCLUSIONS: Many HCV carriers have significant chronic liver damage, although in the majority of them liver lesions are minimal or mild. Up to 60% of HCV carriers in Italy harbour non-1 HCV types. Current definition of HCV carriers with persistently normal ALT levels, based upon three normal ALT values over a 6-month period, is not adequate to discriminate between carriers with persistent ALT normality and those with transient biochemical remission. Longer follow-ups are needed. PMID- 12076872 TI - Polymerase chain reaction-based prevalence of hepatitis A, hepatitis E and TT viruses in sewage from an endemic area. AB - BACKGROUND/AIMS: Hepatitis A and E viruses (HAV, HEV) are transmitted enterically and are highly endemic in India. This study aims to evaluate prevalence of these and TT virus (TTV) in the sewage. METHODS: Influent and effluent samples from a sewage treatment plant from Pune, India were collected twice a week for 1 year and subjected to nested polymerase chain reaction (PCR) for the detection of HAV RNA, HEV RNA and TTV DNA. HAV and HEV PCR products were sequenced. Effluent samples were not collected for 5 months as the plant was non-functional. RESULTS: The overall prevalence was 24.42% (21/86, HAV), 10.98% (9/82, HEV) and 12.7% (8/63, TTV). Prevalence of HAV was significantly higher than HEV (P=0.023). During summer months, significantly higher HAV RNA positivity was noted (P<0.01). A substantial reduction in HAV RNA positivity (15/48 vs. 2/48, P=0.0008) was recorded for treated sewage samples. However, HEV RNA or TTV DNA positivity did not reduce significantly. Of the 17 HAV and HEV RNA negative sewage samples concentrated using ultracentrifugation, 13 and none were positive for HAV and HEV RNA, respectively. Phylogenetic analyses grouped these viruses in IB and Ia, respectively, the genotypes most prevalent in India. CONCLUSIONS: Sewage may play an important role in maintaining hyper-endemicity of these infections. Sustained efforts are obligatory to render sewage less/non-infectious. PMID- 12076871 TI - Early hepatitis C viral kinetics correlate with long-term outcome in patients receiving high dose induction followed by combination interferon and ribavirin therapy. AB - BACKGROUND/AIMS: The majority of patients with genotype 1 do not respond to interferon (IFN) plus ribavirin. Limited data exist on the use of induction followed by combination therapy. METHODS: In this prospective study of 28 patients infected with genotype 1, randomization involved either daily or twice daily high dose IFN for 6 weeks, followed by standard therapy of 3 million units three times a week in combination with ribavirin for an additional 42 weeks. Hepatitis C virus (HCV) RNA was quantitated before and frequently during treatment. RESULTS: The best correlate of response was delta (the infected cell loss rate). Sixteen patients continued on the study because they had at least a 2 log drop in their HCV RNA levels by week 12; all but one were PCR negative for HCV RNA at 48 weeks, and 14 of these 16 patients continued to be PCR negative at 72 weeks. Both African-Americans in our trial failed to respond to therapy, and differences were evident during the induction phase. CONCLUSIONS: This randomized study of induction IFN therapy followed by combination IFN plus ribavirin yielded the highest rate of sustained response (50%) reported to date in chronically HCV infected patients with genotype 1. The predictive value of the infected cell loss rate needs to be evaluated prospectively in larger studies, particularly in patients receiving pegylated IFN. PMID- 12076873 TI - Viral dynamics during and after entecavir therapy in patients with chronic hepatitis B. AB - BACKGROUND/AIMS: Nucleoside analogues inhibit hepatitis B virus (HBV) replication. Entecavir, a new guanine nucleoside, has also been shown to reduce covalently closed circular DNA (cccDNA) to undetectable levels in woodchucks chronically infected with hepatitis virus. Mathematical description of changes in viral load during and after therapy may help to understand the several events that take place during nucleoside analogue treatment. METHODS: Ten chronic hepatitis B patients were evaluated with a mathematical model during and after withdrawal of four doses of entecavir. Blood was drawn for HBV DNA measurement at frequent intervals. Non-linear modelling was used to fit individual patient data. RESULTS: The median effectiveness in blocking viral production is 96% (n=10, range 87-98%). The median half-life of viral turn-over was 16 h (range 12-29 h). The median half-life of infected hepatocytes was 257 h (=10.7 days) (n=9, range 112-762 h). Rebound of viral replication also followed a bi-phasic return to baseline levels. CONCLUSIONS: Decay and rebound of viral concentration during and after entecavir therapy, respectively, showed a bi-phasic pattern. Both can be described with a mathematical model. Data on levels of cccDNA in the liver in these patients could be helpful in supporting the parameters as calculated with the model. PMID- 12076874 TI - Towards a bank of cryopreserved hepatocytes: which cell to freeze? PMID- 12076875 TI - It could have been something they ate -- functional food and the treatment of liver cancer. PMID- 12076876 TI - Favourable and disappointing lessons from viral kinetics. PMID- 12076877 TI - Hepatocellular carcinoma in patients with non-alcoholic steatohepatitis. AB - We describe six patients with non-alcoholic steatohepatitis (NASH) and hepatocellular carcinoma (HCC). From 1990 to 2001, we treated 82 patients with NASH and observed six patients (three men and three women, aged 56-72 years) in this group who were referred with HCC or developed the complication during follow up. In five of these six patients, NASH was associated with obesity (cases 3, 4 and 5), hyperlipidemia (case 5), or diabetes mellitus (cases 1, 3 and 6). We confirmed the presence of HCC by ultrasonography-guided tumor biopsy or surgery except in case 3 where we diagnosed the tumor by ultrasonography, computed tomography and selective hepatic arteriography. The carcinomas measured 1.5-6.0 cm in diameter and three were well differentiated. When HCC was diagnosed, cirrhosis was present in all instances. Four of the six tumor patients also had esophageal varices but only one patient had a history of variceal bleeding and ascites. Treatment of HCC consisted of surgery (cases 1 and 5), transcatheter arterial embolization or infusion and/or percutaneous ethanol injection (cases 2, 3, 4, and 6). In patients with NASH cirrhosis, the development of treatable HCC is sufficiently common to warrant regular screening for this grave complication. PMID- 12076878 TI - Post-splenectomy pancreatic pseudocyst and extensive thrombosis of the portal venous system in a liver transplant recipient. PMID- 12076879 TI - Hyperemesis gravidarum and severe liver enzyme elevation. PMID- 12076880 TI - Acute viral hepatitis A and parvovirus B19 infections complicated by pure red cell aplasia and autoimmune hemolytic anemia. PMID- 12076881 TI - Acute cholestatic hepatitis after reinitiating treatment with atorvastatin. PMID- 12076882 TI - Vapreotide in variceal bleeding. PMID- 12076883 TI - Fracture risk in patients with celiac Disease, Crohn's disease, and ulcerative colitis: a nationwide follow-up study of 16,416 patients in Denmark. AB - The authors studied fracture risk among 16,416 Danish patients with bowel disease. All patients diagnosed with celiac disease (n = 1,021), Crohn's disease (n = 7,072), or ulcerative colitis (n = 8,323) in Denmark between January 1, 1983, and December 31, 1996, were included. Each patient was compared with three age- and gender-matched controls randomly drawn from the background population. No increase in fracture risk could be demonstrated for celiac disease before or after diagnosis. In patients with Crohn's disease, overall fracture risk was increased both before diagnosis (incidence rate ratio = 1.15, 95% confidence interval (CI): 1.00, 1.32) and after diagnosis (incidence rate ratio = 1.19, 95% CI: 1.06, 1.33). Bowel surgery was associated with a decreased risk of sustaining a fracture before diagnosis (odds ratio = 0.70, 95% CI: 0.54, 0.90) and after diagnosis (hazard ratio = 0.81, 95% CI: 0.67, 0.99). Overall fracture risk was not increased in patients with ulcerative colitis, except for a small increase around the time of diagnosis. Increasing age and having a fracture before diagnosis increased the risk of sustaining a new fracture after diagnosis. Crohn's disease was associated with a minor increase in overall fracture risk in contrast to ulcerative colitis and celiac disease. The severity of the inflammatory process and the amount of corticosteroids given may explain the difference in fracture risk. PMID- 12076884 TI - All-cause and cause-specific death rates by educational status for two million people in two American Cancer Society cohorts, 1959-1996. AB - Low socioeconomic status is associated with high mortality, but the extent to which socioeconomic status affects particular diseases and whether socioeconomic status effects have changed over time are uncertain. The authors used education as a marker for socioeconomic status in a study of two large American Cancer Society cohorts (follow-up, 1959-1996). Low education was associated with higher death rates in both cohorts from all causes and most specific causes, except breast cancer and external causes among women. Life expectancy in the more recent cohort was 4.8 years shorter for men and 2.7 years shorter for women for the least versus the most educated. The inverse relation between education and mortality was strongest for coronary heart disease, lung cancer, diabetes, and chronic obstructive pulmonary disease; moderate for colorectal cancer, external causes (men only), and stroke; weak for prostate cancer; and reversed for external causes among women. The direction of a weak gradient for breast cancer differed for those with and without prevalent breast cancer at baseline. Adjustment for conventional risk factors, probable intermediate variables between education and mortality, diminished but did not eliminate the observed educational/mortality gradients. Temporal trends showed increasing mortality differences by education for coronary heart disease, diabetes, and lung cancer for women. PMID- 12076885 TI - Dietary fat intake and ovarian cancer in a cohort of US women. AB - Several studies have suggested that high intake of fats and fat-rich foods may increase the risk of ovarian cancer. The authors examined these relations in the Nurses' Health Study cohort. Dietary intake was assessed in 1980, 1984, 1986, and 1990 by using a self-administered food frequency questionnaire. Food data were used to calculate intake of various fats and fatty acids. For best reflection of long-term intake, an updated, cumulative, averaged measure of fat intake was used to predict incidence of ovarian cancer. Between 1980 and 1996, 301 incident cases of invasive epithelial ovarian cancer were confirmed among the 80,258 participants who completed the baseline food frequency questionnaire. There was no evidence of a positive association between intake of any type of fat and ovarian cancer risk, even after adjustment of fat subtypes for one another. Women in the highest quintile of total fat intake were not at increased risk compared with those in the lowest quintile (multivariate relative risk = 1.03, 95 percent confidence interval: 0.72, 1.45, p for trend = 0.97). Intakes of fat-rich foods were also not appreciably associated with ovarian cancer risk, although an increase in risk with frequent intake of eggs was observed. Overall, results suggest no association between intake of any type of fat and ovarian cancer. PMID- 12076886 TI - Effects of climate, latitude, and season on the incidence of Bell's palsy in the US Armed Forces, October 1997 to September 1999. AB - Bell's palsy is a relatively common disease characterized by the sudden onset of unilateral facial paralysis. Using a centralized surveillance system that contains demographic, military assignment, and medical encounter data of US military service members, the authors estimated rates, trends, and demographic correlates of risk of Bell's palsy during a 2-year period. Poisson regression was used to estimate the independent effects of climate, season, and latitude. From October 1997 to September 1999, there were 1,181 incident cases of Bell's palsy among US service members. The crude incidence rate was 42.77 per 100,000 person years. Incidence rates increased with age and were higher among females, Blacks, Hispanics, married persons, and enlisted service members. Both climate (adjusted rate ratio for arid vs. nonarid climate = 1.34) and season (adjusted rate ratio for cold vs. warm months = 1.31) were independent predictors of risk of Bell's palsy. Latitude was not a statistically significant predictor when demographic, climate, and season effects were taken into account. The results are consistent with hypotheses regarding viral etiologies (e.g., reactivation of herpes simplex) of Bell's palsy. PMID- 12076887 TI - Epidemiology of inapparent and symptomatic acute dengue virus infection: a prospective study of primary school children in Kamphaeng Phet, Thailand. AB - Dengue viruses are a major cause of morbidity in tropical and subtropical regions of the world. Knowledge about the epidemiology and host determinants of inapparent and severe dengue virus infections is limited. In this paper, the authors report findings from the first 3 years of a prospective study of dengue virus transmission and disease severity conducted in a cohort of 2,119 elementary school children in northern Thailand. A total of 717,106 person-school days were observed from 1998 to 2000. The incidence of inapparent and of symptomatic dengue virus infection was 4.3% and 3.6% in 1998, 3.2% and 3.3% in 1999, and 1.4% and 0.8% in 2000, respectively. Symptomatic dengue virus infection was responsible for 3.2%, 7.1%, and 1.1% of acute-illness school absences in 1998, 1999, and 2000, respectively. The early symptom complex of acute dengue virus infection is protean and difficult to distinguish from other causes of febrile childhood illnesses. The authors' results illustrate the spatial and temporal diversity of dengue virus infection and the burden of dengue disease in schoolchildren in Thailand. Their findings increase understanding of dengue virus transmission and disease severity in a well-defined cohort population and offer a study design in which to test the efficacy of potential dengue vaccines. PMID- 12076888 TI - Spatial and temporal circulation of dengue virus serotypes: a prospective study of primary school children in Kamphaeng Phet, Thailand. AB - Dengue virus occurs as four distinct serotypes, each of which causes epidemics throughout the tropical and subtropical regions of the world. Few studies have examined co-circulation of multiple dengue virus serotypes in a well-defined cohort population over time and their capacity to produce severe dengue disease. In this paper, the authors report the details and findings of the first 3 years (1998-2000) of an ongoing prospective study of dengue virus transmission and disease severity in a cohort of children in northern Thailand. A total of 108 dengue virus isolates were obtained from 167 acute dengue virus infections; 23% were DEN-1, 35% were DEN-2, 41% were DEN-3, and 1% were DEN-4. Despite the proximity of the schools, there was marked spatial and temporal clustering of transmission of each dengue serotype. Serotype-specific antibody levels prior to the dengue transmission season were not predictive of the incidence of dengue virus infections or the predominant serotype transmitted at individual schools. All dengue serotypes produced severe dengue illness, although DEN-3 produced more severe symptoms than the other dengue serotypes. The authors' findings emphasize the complexity of dengue serotype-specific virus transmission and severe dengue disease and have important implications for dengue control and vaccine development. PMID- 12076889 TI - Commentary: what contributes to a successful career in epidemiology in the United States? AB - The authors conducted a study examining perceived enabling factors and barriers to a successful career in epidemiology, the role of mentoring in facilitating one's career, where graduates are most often being employed, and key competencies for future epidemiologic training. During June to August 2001, they surveyed senior epidemiologists across the United States (n = 248) in four sectors: state health departments, the Centers for Disease Control and Prevention, the National Institutes of Health, and schools of public health. The top enabling factors were dedication to hard work and having an intrinsic curiosity and a sense of discovery. The most frequently cited barrier was balancing career and family life, except among minority respondents, for whom an unsupportive supervisor was the leading obstacle. Influential characteristics of a mentor were high integrity and the provision of inspiration and encouragement. The top competencies anticipated for the next 10 years were skills working in multidisciplinary teams and in using modern information technologies. Important competencies varied somewhat according to work sector. These findings may be useful in training and career planning among aspiring epidemiologists and for educational policy development among organizations promoting training and mentoring. PMID- 12076890 TI - Using a telephone survey to acquire genetic and behavioral data related to cigarette smoking in "made-anonymous" and "registry" samples. AB - In the Smokers and Nonsmokers Study, the authors investigated the feasibility of using random digit dialing telephone interviews to locate adults in the continental United States who were willing to provide DNA from buccal swabs through the mail. Interviews with 3,383 adults regarding their smoking-related behaviors (response rate = 70%) were conducted in 1999-2000; swab returns continued into early 2001. Overall, 57% of interviewees agreed to receive mailed information explaining the study. Better-educated persons (odds ratio (OR) = 1.3, 95% confidence interval (CI): 1.1, 1.6), younger persons (OR = 0.988, 95% CI: 0.983, 0.992), persons with symptoms of depression (OR = 1.8, 95% CI: 1.4, 2.4), and current smokers (OR = 2.25, 95% CI: 1.8, 2.8) were likelier to agree to receive a mailing. Approximately 26% of interviewees (45% of those receiving kits) returned buccal swabs, and 18% were successfully genotyped. Older (OR = 1.02, 95% CI: 1.01, 1.03), better-educated (OR = 1.4, 95% CI: 1.1, 1.7), and White (OR = 1.8, 95% CI: 1.4, 2.5) participants were more likely to return DNA samples, but current smokers (OR = 0.6, 95% CI: 0.5, 0.8) were less likely to do so. Participants were randomly assigned to one of two forms of participation: the "registry" group (names were kept on file) or the "made-anonymous" group (names were unassociated with samples). The two groups were equally likely to return kits, but registry respondents were more likely to nominate siblings for participation in the study (OR = 1.6, 95% CI: 1.2, 2.1). The participants in this study were similar demographically to the national population. The authors conclude that random digit dialing surveys coupled with mail collection of DNA may constitute a practical method of obtaining DNA samples for biobehavioral research. PMID- 12076891 TI - Agreement on nutrient intake between the databases of the First National Health and Nutrition Examination Survey and the ESHA Food Processor. AB - The objective of this study was to assess agreement on nutrient intake between the nutrient database of the First National Health and Nutrition Examination Survey (NHANES I) and an up-to-date (December 1998) nutrient database, the ESHA Food Processor. Analysis was conducted among 11,303 NHANES I participants aged 25 74 years in 1971-1975 who had undergone dietary assessment. A list of all unique foods consumed was obtained from a single 24-hour dietary recall questionnaire administered during the baseline NHANES I visit. Foods on the list were matched to foods in the ESHA Food Processor software. Agreement between participants' nutrient intakes as calculated with the NHANES I and ESHA nutrient databases was assessed using intraclass correlation analysis, linear regression analysis, and graphic methods. Intraclass correlation analysis demonstrated excellent concordance between most nutrient intakes, with coefficients above 0.95 for intakes of energy, carbohydrates, protein, cholesterol, and calcium; coefficients between 0.90 and 0.95 for intakes of total fat, saturated fat, potassium, and vitamin C; and coefficients of approximately 0.85 for intakes of sodium and vitamin A. Graphic methods and regression analyses also showed good-to-excellent correspondence for most nutrients. These findings support the validity of expanding existing nutrient intake databases to explore current hypotheses, provided that food formulation, enrichment, and fortification practices have not changed substantially over time. PMID- 12076892 TI - Sources of bias in studies among infertility clients. AB - An assumption in case-control studies is that forces of selection are the same for cases and controls. This may not be true for studies of male infertility among infertility clients. Earlier reproductive outcomes may introduce modification of risk behavior or differential referral. Selection bias might also occur when infertile males are compared with fertile males. Partners of sterile men are more likely to have "normal" fertility, while partners of a reference group of normozoospermic men tend to have a lower fertility potential. The latter may lead to overrepresentation of causes of reduced female fertility and introduce bias into estimates of risk factors shared by couples. The relation between cigarette smoking and semen quality was studied in a population of infertility clients from the Netherlands during 1995-1996. To reduce the potential for bias, this relation was studied first in a restricted population less aware of the type of infertility involved. The odds ratio of infertility with smoking was elevated in the restricted population as compared with the total population. Adjustment for smoking by the female partner increased the odds ratios for male smoking as well. These results indicate that bias may occur in clinic-based fertility studies because of different forms of selection. PMID- 12076894 TI - Imaging in the diagnosis, staging, and follow-up of colorectal cancer. PMID- 12076896 TI - Use of the American College of Radiology BI-RADS guidelines by community radiologists: concordance of assessments and recommendations assigned to screening mammograms. AB - OBJECTIVE: This study evaluated the use of the American College of Radiology Breast Imaging Reporting and Data System (BI-RADS) by community radiologists by determining the concordance of assessment categories and recommendations assigned to screening mammograms. MATERIALS AND METHODS: The study comprised the interpretations of 82,620 consecutive screening mammograms by 18 radiologists between January 1, 1995, and December 31, 1998. For all mammograms, assessment categories and recommendations were compared to determine whether they were in accordance with BI-RADS guidelines. Overall patterns of discordance were analyzed, and comparisons of discordant patterns by assessment category, patient age, breast density, and year of examination were made. RESULTS: The overall discordance between BI-RADS assessments and recommendations was low (3%). The assessment with the highest discordance was "probably benign finding" (category 3), at 53.5%. Mammograms obtained in 1998 were almost half as likely to have assessment-recommendation discordance compared with those obtained in 1995 (2.4% vs 4.5%, respectively; odds ratio = 0.52; p < 0.001). Mammograms of women with dense breast tissue were 30% more likely to have lesions assigned discordant assessments and recommendations compared with those of women with fatty tissue (3.4% vs 2.7%, respectively; odds ratio = 1.3; p < 0.001). No differences in the patterns of discordance were found between mammograms of women younger than 50 years and those of women 50 years old and older (p = 0.10). CONCLUSION: There has been improvement in the accurate application of BI-RADS since its introduction. However, variation in the pairing of BI-RADS assessments and recommendations persists. Continued efforts to educate radiologists about the use of BI-RADS and to clarify BI-RADS terms would promote maximum consistency in this use of this reporting method. PMID- 12076897 TI - Compliance with prompt payment legislation: the initial experiences of New Jersey radiologists. AB - OBJECTIVE: Prompt payment has emerged as a dominant issue in managed care reform, with 47 states passing laws or regulations requiring prompt remuneration for medical services. The New Jersey Prompt-Pay Act, effective on December 28, 1999, requires payment within 30 days of electronic submission and within 40 days for nonelectronic submission. This study was undertaken to assess compliance with the New Jersey Prompt-Pay Act for radiology claims submitted 1 and 5 months after implementation (7-11 months after passage of the statute). MATERIALS AND METHODS: A retrospective prompt payment analysis was undertaken of all claims submitted in February and June 2000 to six major payers by a common third-party billing company on behalf of 11 radiology practices of various sizes, settings, and locations in New Jersey. A total of 33,537 claims were assigned to one of six time periods on the basis of timeliness of payment: less than or equal to 30 days, 31-40 days, 41-50 days, 51-60 days, 61-90 days, and greater than 90 days or a separate unpaid category after 170 days had elapsed. A detailed analysis of 3156 claims from one practice was performed to estimate a clean claims submission rate. RESULTS: The overall percentage of claims paid within 40 days was 70%. After 170 days, the overall percentage of paid claims was 93%. The annualized interest lost on delayed payments was $23,939 for the practice analyzed. CONCLUSION: Prompt payment for radiology services remains an elusive goal in New Jersey, despite passage and implementation of prompt payment legislation. PMID- 12076899 TI - Technologists' productivity when using PACS: comparison of film-based versus filmless radiography. AB - OBJECTIVE: The objective of this study was to assess the impact of filmless operation and computed radiography on technologists' examination times compared with conventional film-based operation and film-screen radiography. CONCLUSION: Compared with conventional film-screen operation, filmless operation using computed radiography was associated with a significant decrease in technologist examination times in the performance of general radiographic examinations. This decrease in technologist examination times in a filmless environment offers the potential for increased productivity with resulting personnel savings and improved operational efficiency. PMID- 12076898 TI - Body MR imaging and CT volume: variations and trends based on an analysis of medicare and fee-for-service health insurance databases. AB - OBJECTIVE: We examined Medicare and fee-for-service data sets to understand better the utilization of MR imaging for imaging the pelvis, abdomen, and chest relative to its use in imaging for other body parts and to the utilization of CT. MATERIALS AND METHODS: CT and MR imaging procedure volumes for pelvis, abdomen, chest, and total were extracted from the 1993, 1996, and 1999 Health Care Financing Administration Physician/Supplier Procedure Summary Master Files, based on CPT-4 codes. We also analyzed a fee-for-service health insurance database for January 1998 through July 1999 from a single northeastern state, which included provider location (rural, suburban, or urban) and type (teaching or nonteaching site). RESULTS: The greatest 3-year Medicare increase was for abdominal MR imaging, from 1996 to 1999 (101% increase). However, pelvic, abdominal, and chest MR imaging together remained less than 5% of total MR imaging. Abdominal MR imaging increased more than did total MR imaging in all 10 Health Care Financing Administration regions. In the fee-for-service database, the relative procedure volume of abdominal MR imaging varied approximately fivefold from rural to urban provider locations, and approximately double from nonteaching to teaching hospitals. CONCLUSION: Although far more abdominal CT than abdominal MR imaging is performed, the rate of abdominal MR imaging utilization has increased more rapidly since 1993. The relative procedure volume of abdominal MR imaging varied more than fivefold from rural to urban provider locations and double from nonteaching to teaching hospitals. PMID- 12076901 TI - CT and computed radiography: the pictures are great, but is the radiation dose greater than required? PMID- 12076903 TI - Relying on the radiologist. PMID- 12076904 TI - Statistically engineering the study for success. PMID- 12076905 TI - Using multidetector CT for preoperative vascular evaluation of liver neoplasms: technique and results. AB - OBJECTIVE: The purpose of our study was to evaluate the performance of CT angiography using multidetector CT (MDCT) for preoperative vascular evaluation in candidates who were scheduled for liver neoplasm resection. SUBJECTS AND METHODS: Forty-two consecutive subjects with malignant liver tumors scheduled for resection were studied with multiphase MDCT. The first 22 subjects underwent both multiphase MDCT angiography and catheter angiography before surgery. The subsequent 20 subjects underwent only preoperative CT angiography. Postprocessing was performed, and the images were analyzed for the depiction of arterial, portal vein, and hepatic vein anatomy and for the identification of important vascular variants. The postprocessing findings were compared and correlated with the findings from catheter angiography (22/42) or intraoperative sonography (42/42) and surgery (42/42). RESULTS: Arterial anomalies were detected on the images of 17 of 42 patients, including a replaced right hepatic artery in five, replaced left hepatic artery in six, accessory right and left hepatic arteries in two, common trunk for the celiac and superior mesenteric arteries in one, and early bifurcation of the celiac artery in one. In 22 patients in whom catheter angiography confirmation was available, the number of arteries and almost all the significant anomalies were correctly identified on CT angiography (accuracy, 97%; sensitivity, 94%; specificity, 100%). In the subset of 20 patients who underwent MDCT angiography without catheter angiography confirmation, all clinically relevant information was provided by CT angiography. The portal and hepatic vein anatomy and the relationships of the liver tumors to the neighboring venous structures were shown on CT. CONCLUSION: Multidetector CT provides valuable preoperative information about hepatic vascular architecture and can be used as a noninvasive alternative to catheter angiography before oncologic liver surgery. PMID- 12076906 TI - Multidetector CT: diagnostic impact of slice thickness on detection of hypervascular hepatocellular carcinoma. AB - OBJECTIVE: The objective of our study was to evaluate the diagnostic impact of varying slice thickness on multidetector CT to optimize detection of hypervascular hepatocellular carcinoma. MATERIALS AND METHODS: Forty-three patients with 87 hypervascular hepatocellular carcinomas (diameter: range, 3-80 mm; mean, 22 mm) and 19 patients with either chronic hepatitis or liver cirrhosis and without hepatocellular carcinoma who had undergone early arterial and late arterial phase imaging of the entire liver on multidetector CT were retrospectively enrolled in this study. The detector row configuration was 2.5 x 4 mm, the pitch was 6, and the scanning time was 10.5 sec for each phase. All patients received contrast medium (2 mL/kg of body weight) at a rate of 5 mL/sec; the mean scanning delay for the early arterial phase was 19.0 sec, and the mean delay for the late arterial phase was 34.5 sec. Eighty 2.5-mm-thick reconstruction images, forty 5-mm-thick reconstruction images, and twenty-six 7.5 mm-thick reconstruction images were obtained for each phase. Each image set was interpreted separately by three observers to detect hypervascular hepatocellular carcinoma by viewing images on a workstation monitor. Sensitivity, positive predictive value, and area under the receiver operating characteristic curve (A(z)) were calculated. We used retrospectively excellent follow-up and imaging or pathologic proof as the gold standard. RESULTS: The mean sensitivity and positive predictive value for hypervascular hepatocellular carcinoma were 76% and 69% on 2.5-mm images, 73% and 69% on 5-mm images, and 67% and 76% on 7.5-mm images, respectively. No significant difference in sensitivity among the images was detected, except by one observer who reported a significant difference in the sensitivity between 2.5- and 7.5-mm images (p < 0.05) and between 5- and 7.5-mm images (p < 0.05). The mean A(z) values were 0.79, 0.80, and 0.78 for 2.5-, 5-, and 7.5-mm images, respectively. No significant difference in A(z) values among the images obtained with different slice thicknesses was detected. CONCLUSION: For multidetector CT identification of hypervascular hepatocellular carcinoma, we found little or no advantage in reducing slice thickness to less than 5 mm. PMID- 12076907 TI - Detection of hepatocellular carcinoma: value of adding delayed phase imaging to dual-phase helical CT. AB - OBJECTIVE: The purpose of our study was to determine the value of adding delayed phase imaging to dual-phase helical CT for the detection of hepatocellular carcinoma. SUBJECTS AND METHODS: One hundred thirteen patients with 131 hepatocellular carcinomas underwent triple-phase helical CT. The diagnosis was established by pathologic examination after surgical resection in all patients. For triple-phase helical CT, hepatic arterial, portal venous, and delayed phase scanning began 30, 60, and 180 sec, respectively, after the injection of 120 mL of iodinated contrast material. Dual-phase helical CT excluding delayed phase and triple-phase helical CT images were reviewed independently by three radiologists on a segment-by-segment basis. Diagnostic accuracy was assessed using receiver operating characteristic analysis in 330 resected segments. Sensitivities and specificities were calculated. The value of the delayed phase images in the characterization of hepatocellular carcinoma was also assessed. RESULTS: The diagnostic accuracy of triple-phase helical CT including delayed phase (area under the curve [A(z)], 0.973) was significantly higher than that of dual-phase helical CT (A(z), 0.954). The mean sensitivity of triple-phase CT (89%) was also significantly higher than that of dual-phase CT (86%). The mean specificities of triple-phase CT (99%) and dual-phase CT (99%) were equal. Delayed phase images were helpful in the characterization of hepatocellular carcinoma in 14% of patients. CONCLUSION: The addition of delayed phase imaging to dual-phase helical CT is valuable for the detection and characterization of hepatocellular carcinoma. PMID- 12076908 TI - Sonographic detection of hepatocellular carcinoma and dysplastic nodules in cirrhosis: correlation of pretransplantation sonography and liver explant pathology in 200 patients. AB - OBJECTIVE: The objective of this study was to determine the sensitivity and specificity of sonography as an aid in detecting hepatocellular carcinomas and dysplastic nodules using explantation correlation in patients with cirrhosis and no known hepatocellular carcinomas. MATERIALS AND METHODS: The sonography reports of 200 patients with cirrhosis who underwent sonography and then underwent liver transplantation within 90 days were retrospectively reviewed for focal solid liver lesions. All focal solid masses detected on sonography were considered possible hepatocellular carcinomas. The sonographic findings were compared with thin-section explanted liver pathologic results. RESULTS: Twenty-seven patients (13.5%) had hepatocellular carcinoma at explantation, including four patients with diffuse, multifocal tumors. Eight of the 39 lesions were detected on sonography for a patient sensitivity of 29.6% and a lesion sensitivity of 20.5%. Sonography revealed three (75%) of four hepatocellular carcinomas larger than 5 cm in diameter, one (50%) of two hepatocellular carcinomas with diameters of 3.1 5.0 cm, one (20%) of five hepatocellular carcinomas with diameters of 2.1-3.0 cm, three (13.6%) of 22 hepatocellular carcinomas with diameters of 1-2 cm, and no lesions with diameters smaller than 1 cm. Forty-two patients (21%) had a total of 126 dysplastic nodules including two patients with innumerable lesions. Sonography depicted only two dysplastic nodules, for a patient sensitivity of 4.8% and a lesion sensitivity of 1.6%. The overall specificity of sonography for either hepatocellular carcinomas or dysplastic nodules was 96%. CONCLUSION: Sonography has low sensitivity but high specificity in revealing hepatocellular carcinomas and dysplastic nodules in patients with a cirrhotic liver requiring liver transplantation. In these patients, sonography should not be the sole imaging modality used for lesion detection before transplantation. PMID- 12076909 TI - Hemodynamic characterization of focal nodular hyperplasia using three-dimensional volume-rendered multidetector CT angiography. AB - OBJECTIVE: The goal of this study was to show the ability of three-dimensional multidetector CT angiography to display the angioarchitecture of focal nodular hyperplasia. CONCLUSION: CT angiography with volume rendering shows the anomalous feeding artery and hepatic draining veins that are characteristic of focal nodular hyperplasia. These features may be helpful in distinguishing focal nodular hyperplasia from other lesions. PMID- 12076911 TI - Gadolinium-ethoxybenzyl-diethylenetriamine pentaacetic acid as an intrabiliary contrast agent: preliminary assessment. AB - OBJECTIVE: We assessed the added efficacy of gadolinium-ethoxybenzyl diethylenetriamine pentaacetic acid (gadolinium-EOB) in depicting biliary structures compared with T2-weighted MR cholangiopancreatography (T2-MRCP) and measured reviewer preference and willingness-to-pay for the added value of biliary contrast. SUBJECTS AND METHODS: Ten patients prospectively underwent T2 MRCP and gadolinium-EOB-enhanced MR cholangiography (EOB-MRC). Three radiologists reviewed the unpaired, then the paired, examinations, rating biliary visualization using a 5-point scale. The common bile, right and left hepatic ducts, and second-order branches were evaluated. Improved biliary visualization using paired over unpaired tests indicated the added value of contrast media. Kappa values measured interobserver reliability. A regression model controlling for fixed effects due to reviewer and subject correlation quantified improvement in ratings attributable to paired review. RESULTS: Average visualization ratings for unpaired review of EOB-MRC were the following: common bile duct, 3.3; right hepatic duct, 2.7; left hepatic duct, 2.5; second-order branches, 1.4. Average visualization ratings for unpaired review of T2-MCRP were the following: common bile duct, 3.4; right hepatic duct, 1.8; left hepatic duct, 2.2; second-order branches, 0.9. Ratings improved using paired tests over EOB-MRC and T2-MRCP for all structures (p < 0.001) except for T2-MRCP common bile duct ratings (p > 0.05). Agreement was moderate to good except for EOB-MRC common bile duct ratings. Paired review improved ratings (chi(2) < 0.0001) over T2-MRCP alone by 1.05 and over EOB-MRC alone by 0.68. Despite significant improvement, reviewers preferred unpaired T2-MRCP (53%) over unpaired EOB-MRC (17%) or paired tests (30%). Reviewers were willing to pay $25 (median) for gadolinium-EOB. CONCLUSION: Combining T2-MRCP and EOB-MRC significantly improved biliary visualization over each test alone. However, improvement was small, and the perceived added value of gadolinium-EOB was modest. PMID- 12076912 TI - Radiofrequency ablation of hepatic tumors: increased tumor destruction with adjuvant liposomal doxorubicin therapy. AB - OBJECTIVE: The purpose of this study was to determine whether the administration of liposomal doxorubicin before radiofrequency ablation increases coagulation more than radiofrequency alone in focal hepatic tumors. SUBJECTS AND METHODS: Fourteen focal hepatic tumors (diameter: mean +/- SD, 4.0+/-1.8 cm) in 10 patients (colorectal cancer, n = 3 patients; hepatocellular carcinoma, n = 4; neuroendocrine tumor, n = 2; breast cancer, n = 1) were treated with internally cooled radiofrequency ablation. In addition to undergoing radiofrequency, five patients (n = 7 lesions) were randomly assigned to receive 20 mg of IV doxorubicin in a long-circulating stealth liposome carrier (Doxil) 24 hr before ablation. Contrast-enhanced helical CT was performed immediately (within 30 min) after radiofrequency ablation (baseline) and 2-4 weeks after ablation. The volume of induced coagulation was measured by three-dimensional reconstruction techniques, and the measurements were compared. RESULTS: For tumors treated with radiofrequency alone, the volume of the thermal lesion had decreased 12-24% (mean +/- SD, 82.5% +/- 4.4% of initial volume) at 2-4 weeks after ablation. By comparison, increased tumor destruction at 2-4 weeks after ablation was observed for all lesions treated with combined Doxil and radiofrequency (p<0.001). Six lesions increased 24-36% in volume, and coagulation surrounding a small colorectal metastasis increased 342%. No coagulation was identified in four unablated control lesions in the two patients receiving Doxil alone. CONCLUSION: Our pilot clinical study suggests that adjuvant Doxil chemotherapy increases tumor destruction compared with radiofrequency ablation therapy alone in a variety of focal hepatic tumors. Optimization of this synergistic strategy may ultimately allow improved clinical efficacy and outcome. PMID- 12076913 TI - Saline injection into the pleural cavity to detect tumors of the hepatic dome with sonography: a new approach for treatment of hepatocellular carcinoma. PMID- 12076915 TI - Using a coaxial technique with a curved inner needle for CT-guided fine-needle aspiration biopsy. PMID- 12076914 TI - Usefulness of imaging-guided catheter drainage and talc sclerotherapy in patients with metastatic gynecologic malignancies and symptomatic pleural effusions. AB - OBJECTIVE: The purpose of our study was to assess the usefulness of imaging guided catheter drainage and talc sclerotherapy in patients with metastatic gynecologic malignancies and symptomatic pleural effusions and to assess the affect of ascites on the success rate of this treatment. MATERIALS AND METHODS: Twenty-five patients (mean age, 63 years) with metastatic gynecologic malignancies who had 26 symptomatic effusions treated at our institution over a 4 year period with imaging-guided catheter drainage and talc sclerotherapy were included in this study. Response to treatment was assessed by comparing pre-, immediate post-, and 30-day postsclerotherapy chest radiographs. Response to the treatment was graded as complete (no reaccumulation), partial (accumulation above immediate post- but below presclerotherapy level), or no response (reaccumulation to or above the presclerotherapy level). The presence of ascites on CT (n = 23), sonography (n = 1), direct intraoperative visualization (n = 1), or at physical examination (n = 1) was also noted. RESULTS: Of the 25 patients, 13 patients with 14 treated malignant effusions survived at least 30 days after sclerotherapy and formed the final study group. The remaining patients either died (n = 11) or were lost to follow-up (n = 1). At 30 days, 12 of the 14 treated effusions showed complete responses and one showed a partial response. The overall response rate was 86%. Abdominal ascites was present at the time of treatment in 11 patients (79%) and did not affect the success rate (p > 0.999). CONCLUSION: Imaging-guided catheter drainage and talc sclerotherapy are an effective treatment for symptomatic pleural effusions in patients with metastatic gynecologic malignancies. Ascites does not adversely affect the response to pleurodesis. PMID- 12076916 TI - Treatment of massive hemoptysis with intraarterial thrombin injection of a bronchial artery. PMID- 12076917 TI - Contrast-enhanced MR angiography of the aorta and lower extremities with routine inclusion of the feet. PMID- 12076918 TI - Periaortitis with ureteral obstruction after endovascular repair of an abdominal aortic aneurysm. PMID- 12076921 TI - Sonography of anorectal, rectal, and perirectal abnormalities. PMID- 12076919 TI - Dynamic MR pancreatography after secretin administration: image quality and diagnostic accuracy. AB - OBJECTIVE: The objective of our study was to assess the improvement of image quality and diagnostic accuracy of secretin-enhanced MR pancreatography compared with conventional MR pancreatography. SUBJECTS AND METHODS: Ninety-five patients were studied with a 1.5-T scanner using a T2-weighted single-slice fast spin-echo sequence. Image quality and diameter of the head, body, and tail portion of the pancreatic main duct, the accessory duct, and the side branches were assessed before and after IV administration of secretin. Diagnoses before and after secretin administration were evaluated in a blinded fashion and correlated to the final diagnoses based on endoscopic retrograde cholangiopancreatography (ERCP), intraoperative results, and clinical follow-up as the reference standard. RESULTS: In patients with a normal pancreatic duct, the visualization of all portions of the main pancreatic duct and the accessory duct was significantly improved with dynamic MR pancreatography (p < or = 0.001). In patients with chronic pancreatitis, the visualization of the main duct was also significantly improved with dynamic MR pancreatography (p < or = 0.05). However, the visualization of the minor duct and the side branches was significantly improved only in patients showing no ductal stricture (p < or = 0.05), compared with those with ductal stricture (not significant). The overall sensitivity for the detection of chronic pancreatitis increased from 77% to 89% using secretin enhanced MR pancreatography. A pancreas divisum was found in eight patients before and 13 patients after secretin administration. The overall negative predictive value of MR pancreatography increased from 84% to 98% after secretin administration. CONCLUSION: Improvement in image quality after secretin stimulation increases the diagnostic value of MR pancreatography in patients with a normal or nondilated main pancreatic duct and may obviate invasive procedures such as ERCP. PMID- 12076922 TI - MR imaging pelvimetry: a useful adjunct in the treatment of women at risk for dystocia? AB - OBJECTIVE: The objective of this study was to test the clinical value of MR imaging for diagnosing cephalopelvic disproportion and for predicting labor outcome in women at risk for dystocia. SUBJECTS AND METHODS: Antepartum fetal sonography and maternal MR imaging pelvimetry measurements were performed at term in 38 pregnant women at risk for dystocia with a single fetus in cephalic presentation. Various methods used to diagnose cephalopelvic disproportion were evaluated in a blinded manner for their accuracy to predict both the presence of cephalopelvic disproportion and the mode of delivery (vaginal vs cesarean). RESULTS: None of the methods tested yielded both high sensitivity (15-100%) and high specificity (24-92%) for determining the presence of cephalopelvic disproportion and high levels of accuracy for predicting labor outcome (overall predictability, 50-74%). CONCLUSION: To achieve increased reliability of MR imaging pelvimetry in the diagnosis and treatment of dystocia and in predicting labor outcome, new methods assessing fetal-pelvic compatibility, including measurements of the pelvic outlet and the shape and configuration of the pelvis, need to be established and prospectively tested before firm recommendations for clinical use can be made. PMID- 12076923 TI - Color-flow Doppler sonography of pseudoaneurysms in patients with bleeding renal angiomyolipoma. PMID- 12076925 TI - Incremental benefit of maximum-intensity-projection images on observer detection of small pulmonary nodules revealed by multidetector CT. AB - OBJECTIVE: Our purpose was to assess the incremental effect of maximum-intensity projection (MIP) image processing on the ability of various observers to detect small (<1 cm in diameter) central and peripheral lung nodules revealed by multidetector CT. MATERIALS AND METHODS: We retrospectively identified 25 patients with metastatic disease, each having from two to nine nodules that were 3-9 mm in diameter. Two senior and three junior reviewers interpreted all images on a workstation. The observers first reviewed axial images (3.75-mm collimation, 3-mm reconstruction interval, multidetector acquisition) in cine and sequential fashion and recorded the size, lobe, and central or peripheral (within 1 cm of the edge of lung) location of each nodule. MIP images (10-mm slab, 8-mm interval) were then reviewed, and additional nodules detected were recorded. Final counts were established by consensus. RESULTS: The reviewers found 122 nodules (71 peripheral, 51 central) in the 25 patients. The addition of MIP slabs significantly enhanced reviewer detection of central nodules (p < 0.001) and junior reviewer detection of peripheral nodules (p < 0.001). MIP slabs also reduced the effects of reviewer experience, particularly for peripheral nodules. CONCLUSION: MIP processing reduces the number of overlooked small nodules, particularly in the central lung. Observer nodule detection remains imperfect even when lesions are clearly depicted on images. PMID- 12076927 TI - Dynamic time-resolved contrast-enhanced two-dimensional MR projection angiography of the pulmonary circulation: standard technique and clinical applications. AB - OBJECTIVE: Time-resolved pulmonary two-dimensional MR projection angiography is a fast acquisition technique that allows the generation of dynamic projection angiograms by a method similar to that used to generate digital subtraction angiograms. MR images are obtained after subtracting the mask defined at the beginning of the sequence from later images, thus generating time-resolved continuous projection angiograms that depict the passage of a bolus through the pulmonary circulation. This article describes the application of this novel technique in three patients with pathologic conditions not previously described with this modality and two control subjects. CONCLUSION: The analysis of the findings on dynamic time-resolved contrast-enhanced two-dimensional MR projection angiography shows that this technique is useful not only in revealing morphologic changes associated with pulmonary disorders but also in following the passage of the bolus through the cardiopulmonary circulation. The latter capability allows qualitative detection of normal or abnormal pathways and thus is potentially of value in the assessment of several pulmonary disorders. PMID- 12076928 TI - Intrapulmonary bronchogenic cyst: CT and pathologic findings in five adult patients. AB - OBJECTIVE: The purpose of our study was to describe the CT and pathologic findings of intrapulmonary bronchogenic cysts in five adult patients. CONCLUSION: Intrapulmonary bronchogenic cysts in adults appear on CT as well-defined ovoid lesions with the attenuation of soft tissue or water, similar to their mediastinal counterparts. Associated with these cysts are areas of mosaic low attenuation and bandlike linear attenuation representing histopathologically confirmed emphysema and bronchiolization or fibrotic change or both in the surrounding lung. PMID- 12076929 TI - Breast lesions detected on MR imaging: features and positive predictive value. AB - OBJECTIVE: The purpose of this study was to analyze features of breast lesions detected on MR imaging that had subsequent biopsy and to determine the positive predictive value (PPV) of these features. MATERIALS AND METHODS: Retrospective review was performed of 100 consecutive solitary MR imaging-detected breast lesions that had MR imaging-guided needle localization and surgical excision. We described lesions, using terms found in a proposed breast MR imaging lexicon. Histologic findings were reviewed. RESULTS: Carcinoma was identified in 25 lesions (25%), including ductal carcinoma in situ (DCIS) in 13 (52%) and infiltrating carcinoma in 12 (48%). Carcinoma was found in 15 (25%) of 60 masses versus 10 (25%) of 40 nonmass lesions; most malignant masses (73%) were infiltrating carcinoma, whereas most malignant nonmass lesions (90%) were DCIS. The features with the highest PPV were spiculated margin (80% carcinoma), rim enhancement (40% carcinoma), and irregular shape (32% carcinoma) for mass lesions and segmental (67% carcinoma) or clumped linear and ductal enhancement (31% carcinoma) for nonmass lesions. Visually assessed kinetic patterns were not significant predictors of carcinoma, but washout was present in 70% of infiltrating carcinomas versus 9% of DCIS lesions (p < 0.01). Carcinoma was present in 17 (19%) of 88 lesions classified as suspicious versus eight (67%) of 12 lesions classified as highly suggestive of malignancy (p = 0.001). CONCLUSION: Among MR imaging-detected breast lesions referred for biopsy, carcinoma was found in 25%, of which half were DCIS. Features with the highest PPV were spiculated margin, rim enhancement, and irregular shape for mass lesions and segmental or clumped linear and ductal enhancement for nonmass lesions. Final assessment categories were significant predictors of carcinoma. PMID- 12076930 TI - MR imaging of mucinous carcinoma of the breast. AB - OBJECTIVE: We examined retrospectively the MR imaging findings in eight patients with mucinous carcinoma of the breast to better describe the imaging characteristics of this disease. CONCLUSION: The MR imaging findings in mucinous carcinomas include dynamic curves of the gradually enhancing type and a very high signal intensity on T2-weighted images compared with other histologic types of invasive ductal carcinoma. These findings appear to be useful for diagnosis. PMID- 12076932 TI - Stereotactic vacuum-assisted breast biopsy: an unusual cause of Mondor's disease. PMID- 12076933 TI - Contrast-enhanced magic-angle MR imaging of the Achilles tendon. AB - OBJECTIVE: The objective of this study was to image the Achilles tendon with MR imaging at the magic angle (the long axis of the tendon at 55 degrees relative to the direction of the main static magnetic field [B(0)]) to detect signal from the tendon, to measure the T1 of the tendon, and to determine patterns of contrast enhancement in control subjects and patients. CONCLUSION: Mean T1 values of 311 +/- 30 msec (at 1.0 T) were found in six volunteers. In six control volunteers, slow uptake of contrast material that dispersed over 40 min-1.5 hr was shown without focal change, with elimination in most cases occurring within 18-24 hr. Small rapidly enhancing focal areas of enhancement were seen next to the insertion of the tendon and centrally within 5-10 min in two control volunteers. The focal areas were located at the sites of the blood supply. A patient with chronic tendinopathy showed early local contrast enhancement that extended widely within the tendon over several hours. Two patients with a partially ruptured or repaired tendon showed marked rapid contrast enhancement. The enhancement was obvious with the tendon at the magic angle but was not evident with the tendon in the usual orientation for MR examinations parallel to B(0). PMID- 12076935 TI - Sterilization of barium for vertebroplasty: an effective, reliable, and inexpensive method to sterilize powders for surgical procedures. PMID- 12076934 TI - Patterns of gadopentetate-enhanced MR imaging of radiocarpal joints of healthy subjects. AB - OBJECTIVE: The aim of our study was to evaluate prospectively the grades and patterns of gadopentetate-enhanced MR imaging in the radiocarpal joints of healthy subjects after IV contrast administration. SUBJECTS AND METHODS: The study included 18 healthy subjects (nine men, nine women; age range, 24-34 years; mean age, 30.8 years). We obtained coronal T1-weighted spin-echo images with fat suppression before and after IV administration of gadopentetate dimeglumine and additional axial T1-weighted spin-echo images after contrast administration. Patterns of signal-intensity enhancement in and around the radiocarpal joints were evaluated qualitatively and quantitatively. RESULTS: In eight (44.4%) of 18 healthy subjects, enhancement of the radiocarpal joints was seen and exclusively located in the region of the prestyloid recess. Enhancement patterns were bandlike in three (16.7%) of 18 healthy subjects, homogeneous in another three (16.7%) of 18, and nodular in two (11.1%) of 18. CONCLUSION: After IV administration of gadopentetate, signal-intensity enhancement in the radiocarpal joint is frequently seen in healthy subjects and is not predictive of inflammatory joint disease. If contrast enhancement is present, three distinct patterns are usually revealed, all invariably located in the region of the prestyloid recess. PMID- 12076936 TI - Graded compression sonography of the colon in the diagnosis of polyps in pediatric patients. AB - OBJECTIVE: We report seven cases of juvenile polyps detected by graded compression gray-scale and color Doppler sonography in five children with nonspecific symptoms. CONCLUSION: Intestinal polyps can be detected by graded compression gray-scale and color Doppler sonography without colonic preparation. On gray-scale sonography, polyps appeared as spherical or ovoid hypoechoic nodules in the colon lumen. Small cysts were identified inside the nodules. Four polyps had fewer and smaller cysts, whereas three others contained many cysts. A hyperechoic layer surrounding the polyp corresponded to the submucosa. In two patients, the polyp was visualized in the transverse colon and caused a colocolic intussusception, which reduced spontaneously during sonography. Color Doppler sonography showed four hypovascularized and three hypervascularized polyps. PMID- 12076937 TI - Epidermoid cyst in a pancreatic accessory spleen mimicking an infected abdominal cyst in a child. PMID- 12076938 TI - Using MR cholangiopancreatography to reveal anomalous pancreaticobiliary ductal union in infants and children with choledochal cysts. AB - OBJECTIVE: The purpose of this study was to determine whether MR cholangiopancreatography can accurately depict anomalous pancreaticobiliary ductal union in children with choledochal cysts. SUBJECTS AND METHODS: Twenty children (age range, 1 month-13 years; mean age, 4.6 years; all girls) who were diagnosed with choledochal cyst by sonography underwent MR cholangiopancreatography with a single-shot fast spin-echo sequence. The type of choledochal cyst and anomalous pancreaticobiliary ductal union were characterized on the basis of MR cholangiopancreatographic findings and were compared with the finding of intraoperative cholangiography. RESULTS: The type of choledochal cyst (type Ia, n = 4; type Ic, n = 7; type IVa, n = 7; type IVb, n = 2) determined on MR cholangiopancreatography correlated with that identified on intraoperative cholangiography in each patient. Anomalous pancreaticobiliary ductal union was detected by MR cholangiopancreatography and intraoperative cholangiography in 12 (60%) and 16 (80%) of 20 patients, respectively. The types of anomalous pancreaticobiliary ductal union as determined on MR cholangiopancreatography (type A, n = 2; type B, n = 7; type C, n = 3) were concordant with those of intraoperative cholangiography in 11 of 12 patients. In five of eight patients with choledochal cyst (type Ia, n = 1; type IVa, n = 5; type IVb, n = 2) in whom MR cholangiopancreatography could not depict anomalous pancreaticobiliary ductal union, anomalous pancreaticobiliary ductal union was documented on intraoperative cholangiography that was performed after choledochal cyst resection. CONCLUSION: MR cholangiopancreatography provides diagnostic information about anomalous pancreaticobiliary ductal union in children with choledochal cyst. PMID- 12076940 TI - Handlebar injury causing pancreatic contusion in a pediatric patient. PMID- 12076939 TI - Children with suspected craniosynostosis: a cost-effectiveness analysis of diagnostic strategies. AB - OBJECTIVE: Our purpose was to evaluate the clinical and economic impact of three evaluation strategies in children at different risks of craniosynostosis. MATERIALS AND METHODS: A decision-analytic and cost-effectiveness model was constructed to compare three evaluation in strategies in children with suspected synostosis: no imaging, radiography (if abnormal, followed by three-dimensional CT [3D CT]), and 3D CT. Three risk groups were analyzed on the basis of the prevalence (pretest probability) of disease: low (completly healthy children; prevalence, 34/100,000), intermediate (healthy children with head deformity; prevalence, 1/115), and high risk (children with syndromic craniofacial disorders [i.e., Crouzon's syndrome or Apert's syndrome]; prevalence, 9-10/10). Test performance (sensitivity and specificity) of the evaluation strategies was obtained from the literature. Costs (not charge) estimates were obtained from the hospital cost-accounting database and from the Medicaid fee schedule. RESULTS: In the low-risk group, the radiographic and 3D CT strategies resulted in a cost per quality-adjusted life year (QALY) gained of more than $560,000. In the intermediate-risk group, the radiographic strategy resulted in a cost per QALY gained of $54,600. Three-dimensional CT was more effective than the two other strategies but at a higher cost-hence, with a cost per QALY gained of $374,200. In the high-risk group, 3D CT was the most effective strategy with a cost per QALY gained of $33,800. Less experienced radiologists and poor-quality studies increased the evaluation cost per QALY gained for all of the risk groups because of decreased effectiveness. CONCLUSION: Radiologic screening of completely healthy children (low risk) for synostosis is not warranted because of the high cost per QALY gained of the radiographic and 3D CT strategies. In healthy children with head deformity (intermediate risk), the radiographic strategy had a reasonable cost per QALY gained. Three-dimensional CT was more effective but had a high cost per QALY gained. In children with syndromic craniofacial disorders (high risk), 3D CT was the most effective strategy and had a reasonable cost per QALY gained. Selection of children with suspected craniosynostosis based on their risk group and use of the most appropriate evaluation strategy could maximize clinical and economic outcomes for these patients. PMID- 12076941 TI - Influence of availability of clinical history on detection of early stroke using unenhanced CT and diffusion-weighted MR imaging. AB - OBJECTIVE: The radiologic diagnosis of stroke requires accurate detection and appropriate interpretation of relevant imaging findings; both detection and interpretation may be influenced by knowledge of the patient's presentation. In our study, we evaluated the effect of the availability of clinical history on the sensitivity for stroke detection using unenhanced CT and diffusion-weighted MR imaging. MATERIALS AND METHODS: The records of 733 consecutive patients with a clinically based admission diagnosis of early stroke were reviewed. Among the criteria for inclusion in our study were the availability of an unenhanced CT scan (561 cases) or diffusion-weighted MR imaging examination (409 cases) obtained at admission and a discharge diagnosis indicating whether a patient had actually had a stroke. The radiology requisition forms, available at the time of image interpretation, were classified as either indicating or not indicating a clinical suspicion of early stroke. Sensitivity, specificity, and accuracy of stroke detection were computed, stratified by the presence or absence of an available history indicating suspicion of stroke. Results were compared using the Fisher's exact two-tailed test. RESULTS: Unenhanced CT sensitivity was 52% (specificity, 95%) for the suspicion-of-stroke group and 38% (specificity, 89%) for the no-suspicion-of-stroke group (p = 0.008). Diffusion-weighted MR imaging sensitivity was 95% (specificity, 94%) for the suspicion-of-stroke group and 94% (specificity, 98%) for the no-suspicion-of-stroke group (p = 0.822). CONCLUSION: Availability of a clinical history indicating that early stroke is suspected significantly improves the sensitivity for detecting strokes on unenhanced CT without reducing specificity. In contradistinction, the availability of such a history did not significantly improve the sensitivity for detecting stroke using diffusion-weighted MR imaging. Whenever possible, relevant clinical history should be made available to physicians interpreting emergency CT scans of the head. PMID- 12076944 TI - Signs of acute stroke seen on fluid-attenuated inversion recovery MR imaging. PMID- 12076942 TI - Quantitative evaluation of mean transit times obtained with dynamic susceptibility contrast-enhanced MR imaging and with (133)Xe SPECT in occlusive cerebrovascular disease. AB - OBJECTIVE: We report on quantitative mean transit time images obtained using dynamic susceptibility contrast-enhanced MR imaging after an IV bolus injection of a contrast agent. Our study compared cerebral hemodynamics measured using dynamic susceptibility contrast-enhanced MR imaging with the cerebral perfusion reserve obtained using dynamic (133)Xe single-photon emission computed tomography (SPECT). SUBJECTS AND METHODS: Seventeen patients with chronic cerebral artery occlusion or stenosis diagnosed by digital subtraction angiography were examined. Dynamic susceptibility contrast-enhanced MR imaging data were acquired using a multishot echoplanar sequence. Our procedure for quantification of mean transit time was based on the indicator dilution theory and deconvolution analysis. RESULTS: The increased mean transit time values obtained with dynamic susceptibility contrast-enhanced MR imaging correlated well (r=-0.789, p < 0.0001) with decreased cerebral perfusion reserve determined by performing dynamic (133)Xe SPECT before and after administration of acetazolamide. The mean transit time values in the regions with severely decreased perfusion reserve were significantly higher than those in the regions with normal or moderately decreased perfusion reserve (p < 0.0001 and p = 0.0004, respectively). CONCLUSION: Mean transit time images generated from dynamic susceptibility contrast-enhanced MR imaging data could be used to evaluate the extent of cerebral perfusion reserve impairment in patients with occlusive cerebrovascular disease. PMID- 12076945 TI - Evaluation of software for registration of contrast-enhanced brain MR images in patients with glioblastoma multiforme. AB - OBJECTIVE: We evaluated commercially available software that rapidly and automatically registers brain MR images on a clinical workstation, and we studied the accuracy of these registrations. SUBJECTS AND METHODS: Ten patients with a diagnosis of glioblastoma multiforme underwent contrast-enhanced inversion recovery prepared three-dimensional (3D) volumetric spoiled gradient-recalled acquisition in the steady state (SPGR) MR imaging (contiguous 1.5-mm slice thickness, 96-104 slices). After this imaging sequence, each patient was brought out of the head coil into a sitting position and then repositioned in the coil. The inversion recovery prepared 3D SPGR sequence was then repeated. A commercially available software program operating on a clinical workstation was used to automatically register the second inversion recovery prepared SPGR series to the first. The speed of registration was recorded. The accuracy of each registration was estimated by recording the coordinates of eight anatomic landmarks on the registered and reference series and by calculating the mean error among matching landmarks. RESULTS: In nine of 10 patients, the registration software produced a visually satisfactory registration. In one patient, a second registration was necessary to produce a satisfactory registration. The processing time for each iteration was 48.3 +/- 3.8 sec (mean +/- SD). The mean error in aligning matching anatomic landmarks ranged from 0.67 to 1.41 mm, with an overall mean of 1.18 mm. The largest error among matching landmarks was 2.3 mm. CONCLUSION: Commercially available registration software can automatically register 3D imaging volumes in less than 1 min. The mean error in registration was approximately equivalent to the dimensions of a single voxel. PMID- 12076946 TI - Lesions of the corpus callosum: MR imaging and differential considerations in adults and children. PMID- 12076947 TI - MR imaging of large multinodular goiters: observer agreement on volume versus observer disagreement on dimensions of the involved trachea. AB - OBJECTIVE: MR imaging and sonography are considered to be among the most reliable methods available for estimating goiter volume. Our aim was to assess the observer variability of MR imaging for estimating the volume of a large multinodular goiter and the dimensions of the related trachea. Additionally, we compared the goiter volume estimates from MR imaging with those from sonography. SUBJECTS AND METHODS: The effect of high-dose (131)I therapy on the thyroid gland and the impact on the trachea in 23 patients with a large multinodular goiter (range in volume, 100-703 mL) were monitored by observers unaware of duplicated measurements on MR imaging (n = 68) before, 1 week after, and 1 year after (131)I therapy. In goiters exclusively cervically located (n = 12), cross-sectional planimetric sonographic measurements (n = 24) were performed simultaneously with MR imaging. RESULTS: The mean intraobserver difference for the MR imaging measurements of goiter volume was 2.1 mL (1.4%, p = 0.32), and the coefficient of variation (CV) +/- SD was 3.6% +/- 2.6%. The mean interobserver difference was 0.4 mL (0.3%, p = 0.86), and the CV +/- SD was 4.1% +/- 3.5%. Compared with MR imaging, sonography underestimated goiter volume; the mean percentage difference between the volume estimates on MR imaging and those on sonography (volume estimated on MR imaging - volume estimated on sonography) was 19.5% (95% limits of agreement: -22.2% to 83.7%), and the CV +/- SD was 15.0% +/- 12.4%. The mean interobserver difference in the MR imaging measurement of tracheal volume along the goiter extension was 7.4% (95% confidence interval: 4.0-10.8%) and that of the smallest cross-sectional area of the trachea was 7.9% (95% confidence interval: 2.9-13.2%). The corresponding CV +/- SD were 8.1% +/- 6.6% and 10.3% +/ 10.3%, respectively. CONCLUSION: For the estimation of goiter volume, MR imaging has low intra- and interobserver variations. In contrast, the determination of tracheal dimensions using MR imaging has a high variability and, thus, is imprecise. Sonography significantly underestimates thyroid volume compared with MR imaging in patients with a large goiter. PMID- 12076948 TI - The palatovaginal canal: can it be identified on routine CT and MR imaging? AB - OBJECTIVE: The palatovaginal canal is a short bone tunnel that extends from the pterygopalatine fossa to the roof of the pharynx. The primary purpose of our work was to establish whether the palatovaginal canal can be identified on CT and MR imaging. The secondary goal was to establish the frequency of visualization and the appearance of this canal. MATERIALS AND METHODS: We retrospectively analyzed 150 consecutive direct coronal CT studies obtained for evaluation of the sinonasal cavities. Frequency, bilaterality, and appearance of the palatovaginal canals were recorded. The frequency of the vidian canals was recorded for comparison. We also analyzed 20 MR imaging studies of that area to assess visualization of the palatovaginal canals and their contents. A dry skull specimen was examined using CT, and the images were correlated with those obtained in vivo. RESULTS: The palatovaginal canal could be identified on CT on at least one side in 88 (58.7%) of 150 patients. Unilateral complete canals were found in 14 patients (9.3%), and unilateral semicanals were evident in 17 (11.3%). Bilateral complete canals were seen in 24 patients (16%), and bilateral semicanals were found in 11 (7.3%). In 22 patients (14.7%), one complete canal and one semicanal were detected. Fifty-five percent of the visualized canals were completely formed. The palatovaginal canal and its internal tubular structure, presumably corresponding to the pterygovaginal artery, were depicted on 40% of the MR imaging studies. The position and configuration of this canal as seen on CT of the dry skull specimen correlated well with the imaging findings. CONCLUSION: The palatovaginal canals are commonly depicted on CT and MR imaging. PMID- 12076949 TI - Are T1- and T2-weighted spin-echo MR images sufficient for the diagnosis of central nervous system vasculitis? PMID- 12076950 TI - Primary amyloidosis of the breast presenting solely as microcalcifications. PMID- 12076951 TI - What is adequate treatment for gastric fundal varices associated with a gastrorenal shunt? PMID- 12076953 TI - Leiomyosarcoma of the renal vein: imaging and surgical reconstruction. PMID- 12076954 TI - Osteosarcoma of the breast. PMID- 12076955 TI - Myelitis caused by Cladophialophora bantiana. PMID- 12076956 TI - Buckshot embolism to the kidney. PMID- 12076957 TI - Biliary adenofibroma with malignant transformation and pulmonary metastases: CT findings. PMID- 12076958 TI - Preconditioning-mediated neuroprotection: role of nitric oxide, cGMP, and new protein expression. AB - Preconditioning adaptation induced by transient ischemia can increase brain tolerance to oxidative stress, but the underlying neuroprotective mechanisms are not fully understood. Recently, we developed a human brain-derived cell model to investigate preconditioning mechanism in SH-SY5Y neuroblastoma cells.(1) Our results demonstrate that a non-lethal serum deprivation-stress for 2 h (preconditioning stress) enhanced the tolerance to a subsequent lethal oxidative stress (24 h serum deprivation) and also to 1-methyl-4-phenyl-pyridinium (MPP(+)).(2) Two-hour non-lethal preconditioning stress increased the expression of neuronal nitric oxide (NOS1/nNOS) mRNA, Fos, Ref-1, NOS protein, and then nitric oxide (*NO) production. As well as MnSOD expression, the *NO-cGMP-PKG pathway mediated the preconditioning-induced upregulation of antiapoptotic protein Bcl-2 and the downregulation of adaptor protein p66(shc). We also propose that cGMP-mediated preconditioning-induced adaptation against oxidative stress may be due to the synthesis of a new protein, such as thioredoxin (Trx) since the protective effect can be blocked by Trx reductase inhibitor.(3) The antioxidative potency of Trx was approximately 100 and 1,000 times greater than GSNO and GSH, respectively. These results suggest that *NO-cGMP-PKG signaling pathway plays an important role in the preconditioning-induced neuroprotection, and perhaps cardioprotection, against oxidative stress. PMID- 12076959 TI - NO-mediated chemoresistance in C6 glioma cells. AB - Expression of inducible nitric oxide synthase (iNOS) in malignant glioma and other tumors has been extensively documented. Massive production of NO by iNOS has been shown to exert tumoricidal effects. However, NO may enhance vasodilation and promote neovascularization, thereby facilitating tumor growth. Compared to the effects of NO on tumor cell death and survival, correlation between NO and cytotoxicity of chemotherapeutic reagents in glioma have been less well characterized. Another gene product often linked to tumor malignancy is hypoxia inducible factor-1 (HIF-1). HIF-1 is a transcription factor that renders malignant tumors adaptive to hypoxic stress during massive vascularization and tumor invasion. Interestingly, HIF-1 also contributes to iNOS induction under hypoxia. We have characterized the interrelationship between iNOS, HIF-1 and chemoresistance. We note that increased NO synthesis by cytokine exposure or iNOS overexpression neutralized the cytotoxicity of 1,3-bis(2-chloroethyl)-1 nitrosourea (BCNU) and 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU), but not cisplatin, in rat C6 glioma cells. Both BCNU and CCNU are chloroethylnitrosoureas that kill tumor cells via carbamoylating and alkylating actions. Further studies indicated that iNOS only neutralized carbamoylating action of chloroethylnitrosoureas. Expression of iNOS may inhibit HIF-1 activity under hypoxia in C6 glioma cells transfected with a VEGF promoter-driven luciferase gene. Pretreatment of C6 cells with N-acetyl-l-cysteine (NAC), an antioxidant, nullified the inhibitory effect of iNOS on HIF-1 binding. That NO generated by iNOS expression inhibits HIF-1 activity in hypoxic C6 cells reveals a negative feedback loop in the HIF-1 --> iNOS cascade. Together these results suggest a complicated role of NO in malignant tumor growth, survival and invasion. PMID- 12076960 TI - Role of nitric oxide in myocardial preconditioning. AB - Ischemic preconditioning (PC) is a polygenic defensive cellular adaptive phenomenon whereby brief ischemic stimuli render the heart resistant to subsequent similar stress. The late phase of ischemic PC lasts for three to four days, protects against both myocardial stunning and infarction, and thus has considerable clinical relevance. Diverse signaling molecules released by a sublethal ischemic stress initiate a complex signal transduction cascade that modulates the expression of cardioprotective genes. Nitric oxide (NO), generated by the endothelial NO synthase (NOS) and acting via the formation of reactive oxygen species, activates the epsilon isoform of protein kinase C (PKC), which activates the Src family of protein tyrosine kinases (Src and Lck) and transcription factors (nuclear factor-kappaB, and possibly others), with resultant upregulation of the inducible NOS (iNOS) gene and protein expression. iNOS, and other cardioprotective proteins, including cyclooxygenase-2 and aldose reductase, confer resistance to subsequent ischemic stress. This delayed protection can also be mimicked, in the absence of ischemia, by administering NO releasing agents, a situation that can be potentially exploited for cardioprotection in clinical situations. Identification of this novel bifunctional (trigger and mediator) role played by NO in cardiac protection, not only advances the knowledge regarding its signaling functions, but also offers a potential therapeutic strategy for patients with coronary artery disease. The purpose of this review is to summarize the current evidence in support of this critical role played by NO in ischemic PC. PMID- 12076961 TI - Regulation of caspases by nitric oxide. AB - Nitric oxide can prevent or induce apoptosis depending on its concentration, cell type, and the oxidative milieu. Nitric oxide inhibits apoptosis and inflammation by S-nitrosylation of the active site cysteine of caspases, the central effector molecules of cell death as well as maturation of IL-1beta and IL-18. The ability of nitric oxide to S-nitrosylate caspases depends on multiple factors including the presence of free iron and intracellular redox potential. There are no known direct effects of nitric oxide on promoting caspase activation or activity. However, nitric oxide has been shown to promote apoptotic pathways in numerous cell types through the indirect activation of caspases. In this article we review the relationship of nitric oxide and caspase activity, modulation of this effect by iron, and clinical implications for the use of nitric oxide in regulating inflammation and apoptosis. PMID- 12076962 TI - The significance of nitric oxide production in the brain after injury. AB - Glutamate toxicity has been implicated in many aspects of brain injury including traumatic, ischemic, and hemorrhagic damage. We have used in vitro as well as in vivo methods to measure NO production and to examine the role of NO in glutamate toxicity. In building our recombinant system, we used human kidney embryonic cells, HEK 293, as host for transfection of nNOS and NMDA receptor proteins. Cells cotransfected with NMDA and nNOS were more resistant to glutamate toxicity. This resistance correlated with NO production as measured by citrulline assay. Meanwhile, the production of NO did not significantly change the response of the NMDA receptor as seen by calcium studies. Moreover, in vivo, NO production was directly correlated with brain tissue oxygen tension in subarachnoid hemorrhage patients. These data and others point toward the importance of NO production in the response of brain to injury. PMID- 12076963 TI - Contradictory effects of sodium nitroprusside and S-nitroso-N-acetylpenicillamine on oxidative stress in brain dopamine neurons in vivo. AB - To investigate whether nitric oxide (*NO) is neurotoxic or neuroprotective in the brain, we compared the in vivo role of S-nitroso-N-acetylpenicillamine (SNAP) with that of sodium nitroprusside (SNP) on ferrous citrate-induced oxidative stress and neuronal loss in the rat nigrostriatal dopaminergic system. It is known that light irradiation releases *NO from its donor compounds; these irradiated *NO donors were used as sham controls in this study. Intranigral infusion of ferrous citrate (4.2 nmol) into the rat midbrain substantia nigra compacta area caused acute lipid peroxidation in the substantia nigra and chronic dopamine depletion in the caudate nucleus. Coinfusion of freshly prepared SNAP (0 8.4 nmol) or *NO (about 2 nmol), but not SNP, rescued iron-induced dopamine depletion in the rat brain in vivo. In fact, SNP produced prooxidative effects similar to ferrous citrate both in vivo and in vitro, since SNP is a redox iron complex. Consistently, *NO and SNAP inhibited, whereas SNP potentiated, *OH generation and lipid peroxidation evoked by ferrous citrate in vitro. We previously reported that freshly prepared, but not irradiated, S-nitroso-L glutathione (GSNO) protected brain dopamine neurons against oxidative stress in vivo. As well as these antioxidative properties, our recent reports (see (Ref. 1)) indicate that *NO/GSNO activated guanylyl cyclase, increased cGMP and that could lead to PKG-mediated expression of MnSOD, Bcl-2, and thioredoxin for preconditioning neuroprotection against 1-methyl-4-phenylpyridinium (MPP(+)).(1) In conclusion, *NO and S-nitrosothiols (e.g., GSNO and SNAP) can scavenge reactive oxygen species and activate the heme moiety of guanylyl cyclase, resulting in protection of brain dopamine neurons through both antioxidative and antiapoptotic mechanisms. PMID- 12076965 TI - Glial-neuronal transfer of arginine and S-nitrosothiols in nitric oxide transmission. AB - The arginine-nitric oxide (Arg-NO) and the S-nitrosothiols systems, two less well studied aspects of NO transmission in the central nervous system, are reviewed. A growing body of evidence suggested that they play a crucial role in NO synthesis and activity. l-Arginine, the NO precursor, is predominantly localized in glia. Together with in vitro and in vivo results of arginine release, this suggests a transfer of arginine from glia to neurons in order to supply NO synthase with its substrate. NO biosynthesis may thus involve the co-occurrence of the glial neuronal transfer of arginine and of NOS activation. The arginine availability may shed light on the dual, beneficial and toxic effects of NO. At low arginine concentrations, neuronal NO synthase generates NO and superoxide, favouring the production of the toxin peroxynitrite. NMDA-induced excitotoxicity in neuronal cells is dependent on arginine availability and glia may play a neuroprotective role by supplying arginine. The reversible S-nitros(yl)ation of thiol containing molecules may represent an important cellular signal transduction mechanism, probably comparable to phosphorylation. S-nitrosothiols, in particular through the presence and release of S-nitroso-cysteinylglycine in sensory thalamus, may act as a local buffering system in NO transmission. This may represent a novel specific facilitating mechanism in order to enhance transmission of persistent stimuli. PMID- 12076964 TI - L-Citrulline recycle for synthesis of NO in cerebral perivascular nerves and endothelial cells. AB - Recycle of L-citrulline to form L-arginine in cerebral perivascular nerves has been well described, providing direct evidence that nitric oxide (NO) is synthesized and released from these nerves to act as the transmitter for vasodilation. NO is also synthesized and released from cerebral endothelial cells, involving L-citrulline conversion to L-arginine. Evidence for the presence of enzymes involved in the conversion, however, has not been shown. The presence of nitric oxide synthase (NOS), argininosuccinate synthetase (ASS), and argininosuccinate lyase (ASL), and their coexistence with NADPH-diaphorase (NADPHd), a marker for NOS, in endothelial cells of middle cerebral arteries and the circle of Willis of the pig, therefore, were examined using combined immunohistochemical and histochemical techniques. NOS-, ASS-, and ASL immunoreactivities were found in almost all endothelial cells of all cerebral arteries examined. All ASS-, ASL-, and NOS-immunoreactive (I) endothelial cells also stained positively for NADPHd, suggesting that ASS, ASL, and NOS were colocalized in endothelial cells of middle cerebral arteries and the circle of Willis. These results provide morphological evidence that cerebral vascular endothelial cells like cerebral perivascular nerves contain enzymes necessary for recycling L-citrulline to L-arginine to synthesize NO via an argininosuccinate (AS) pathway. PMID- 12076966 TI - Nitric oxide protects astrocytes from oxidative stress. AB - Oxidative stress in astrocytes has been observed in a number of pathological conditions, however the effect of nitric oxide on oxidatively stressed astrocytes remains unclear. The goal of this research was to determine how nitric oxide influences the mechanism of peroxide-induced oxidative stress in astrocytes. We used enriched mouse-astrocyte cultures and tertiary-butyl hydroperoxide to study astrocyte oxidative stress. Cellular integrity was measured at the ultrastructural level by electron microscopy and plasma membrane integrity measured by lactate dehydrogenase release. Mitochondrial swelling and disruption of cristae were observed by electron microscopy in the absence of damage to other organelles. Astrocytes exposed to peroxide eventually succumbed to necrotic loss of plasma membrane integrity. However, when astrocytes were subjected to oxidative stress in the presence of nitric oxide, mitochondrial integrity was preserved and cell death delayed. This protection of nitric oxide resembled that of iron chelation and nitric oxide reduced susceptibility usually associated with iron-loaded astrocytes exposed to oxidative stress. Taken together, these data suggest that nitric oxide can act as a powerful antioxidant for astrocytes preserving mitochondrial and cellular integrity during oxidative stress. PMID- 12076967 TI - The roles of nitric oxide in dorsal root ganglion neurons. AB - The functions of nitric oxide (NO) in primary somatosensory neurons are reviewed. During the early development of these neurons the neuronal isoform of nitric oxide synthase (nNOS) is expressed during neurite extension. As their axons extend peripherally and start to take up nerve growth factor (NGF), nNOS starts to disappear from the majority of these neurons. A small number (less than 5%) continue to express nNOS, and for this small population NO may have a role in synaptic transmission. Following peripheral nerve section in adult rats, nNOS is reexpressed in many small peptidergic DRG neurons that have been axotomized. At the same time cGMP synthesis is increased in satellite glia cells. From culture studies, it was established that NGF negatively regulates nNOS synthesis in DRG neurons and that block of NO production leads to neuronal death. Further recent data is reviewed that supports the view that NO has a neuroprotective action preventing loss of DRG neurons and facilitates regeneration. PMID- 12076968 TI - Spatial and temporal correlation of nitric oxide synthase expression with CuZn superoxide dismutase reduction in motor neurons following axotomy. AB - Axotomized neurons expressing neuronal nitric oxide synthase (nNOS) may use nitric oxide (NO), known for its antioxidant activities and ability to scavenge free radicals, to protect against oxidative stress. This hypothesis was tested by immunohistochemical examination of superoxide dismutase (SOD) in neurons of the hypoglossal nucleus (HGN) and dorsal motor nucleus of the vagus nerve (DMV) one day to ten weeks after unilateral hypoglossal nerve crush or avulsion combined with vagus nerve crush in adult rats, and also in neurons of the anterior horn (AH) one week after unilateral sciatic nerve crush or avulsion. In the HGN, emergence of nNOS coincided temporally with reduction of CuZn-SOD immunoreactivity (ir), and the level of reduction correlated with that of nNOS induction, differing only in magnitude between nerve crush and nerve avulsion. The two nerve lesion models further revealed the concurrence of nNOS abatement with recovery of CuZn-SOD ir, and absence of nNOS abatement with persistent low CuZn-SOD ir. In the AH, reduced CuZn-SOD ir was localized in the segments containing nNOS positive neurons as a result of sciatic nerve avulsion. CuZn-SOD ir was unchanged in the absence of nNOS induction following sciatic nerve crush. DMV neurons were devoid of CuZn-SOD ir. However, increased Mn-SOD ir one and two weeks post crush was similar to that in HGN neurons. DMV neurons lacked both nNOS abatement and CuZn-SOD ir, which may explain their particular vulnerability to cell death from axotomy in comparison with other peripheral neurons. These data suggest that axotomy-induced nNOS expression is causally linked to oxidative stress, and that NO is neuroprotective, but can become neurodestructive when produced in excess. PMID- 12076969 TI - Regulation of endothelial nitric oxide synthase activity and gene expression. AB - Endothelial nitric oxide synthase (eNOS) is constitutively expressed in endothelial cells lining the blood vessel and the heart. It plays a major role in vascular and tissue protection. Its activity is tightly controlled by an intramolecular autoinhibitory element that hinders calmodulin binding. This molecular hindrance is removed by elevated intracellular calcium levels. The catalytic activity of eNOS is augmented by phosphorylation of a C-terminal serine residue (Ser-1177 of human eNOS) through the phosphatidyl-3 kinase (PI-3K)/Akt pathway. Its activity is also enhanced by binding to heat shock protein-90. These two processes are calcium independent. The two biochemical events appear to facilitate calmodulin access to its binding site. eNOS is upregulated at the transcriptional level. Its upregulation is mediated by an increased Sp1 binding to its cognate site on eNOS promoter/enhancer region via the action of protein phosphatase 2A (PP2A). PP2A is activated by a signaling pathway including PI 3gamma --> Janus activated kinase 2 (Jak2) --> MEK-1 --> ERK1 and 2. The transcriptional and posttranslational enhancement of eNOS activity is two- to threefold above the basal level. A higher magnitude of augmentation of eNOS gene expression can be achieved by gene transfer, which confers protection against vascular diseases and ischemia-induced tissue injury in experimental animals. These findings provide new insight into the protective role of eNOS and the therapeutic potential of eNOS gene therapy. PMID- 12076970 TI - Nitric oxide transport and storage in the cardiovascular system. AB - Despite short halflife in biological fluids, nitric oxide (NO) can produce remote or long lasting effect in the cardiovascular system. Long distance transport or local storage of NO might explain these effects. In blood, recent findings suggest that in addition to being a major consumption pathway, interaction of NO with hemoglobin may permit O(2)-governed transport of NO (as S-nitrosohemoglobin) to tissues in which NO may be released together with O(2), via transnitrosation of a transport protein. In blood vessels, two different putative NO stores have been characterized. The first is the photosensitive store, formed from endothelium-derived NO. The mechanism of NO release from this store in the body (in absence of light) and its physiological relevance are unknown. The second store is generated in conditions of high tissue NO levels, as a consequence of the inducible NO synthase activity or in various stress conditions. This NO store involves formation of protein-bound dinitrosyl iron complexes or S-nitrosated proteins, or both. Low molecular weight thiols can displace NO from these stores and probably transfer it to target membrane protein(s) such as K(+) channels, via transnitrosation reactions. These stores may be involved in defence mechanisms against inflammation or stress. Thus, NO transport and storage mechanisms may be implicated in a variety of NO effects. The mechanisms of their formation and of NO release and their physiologic and pathophysiologic relevance deserve further investigations. PMID- 12076972 TI - Does nitric oxide play a role in orofacial pain transmission? AB - Although the synaptology, neural connectivity, and the roles played by nitric oxide (NO) and other neurotransmitters have been extensively studied in spinal pain, such information is rather scanty with respect to orofacial pain transmission. This paper presents the findings of several investigations carried out by the author and his colleagues on the roles of NO in orofacial pain transmission in male Wistar rats, using nicotinamide adenosine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry using light and electron microscopy; and NOS immunohistochemistry and immunofluorescence using both light and confocal laser scanning microscopy. The results revealed that (1) a complicated relation existed between the nitrergic axon terminals and dendrites in the caudal part of the spinal trigeminal nucleus (cSTN); (2) the nitrergic neuronal cells bodies were not projection neurons, but rather, local circuit neurons; (3) although the thalamus projecting neurons in the cSTN did not synthesize NO, they could be modulated by NO diffused from nitrergic neurons; (4) c-fos positive neurons in the superficial laminae of the cSTN, detected following subcutaneous injection of 0.5 ml of 4% formalin into the left lateral face of the rats, respond to the release of glutamate through activation of N-methyl-D aspartate (NMDA), alpha-amine-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) and metabotropic glutamate (mGlu) receptors expressed by these c-fos neurons; and (5) NO might play a seemingly less important role than glutamate in neural transmission. PMID- 12076971 TI - Thioredoxin-related regulation of NO/NOS activities. AB - The role of regulation of nitric oxide synthase (NOS) activity in mitigating oxidative stress in neonatal lungs and contributing to pulmonary vasodilation at birth is still unclear. Furthermore, it is known that, depending on interactions between the individual components of the mitogen-activated protein kinase (MAPK) signaling cascades, many biological consequences, including apoptosis, are initiated. Although the importance of nitric oxide (NO) in apoptosis is controversial and likely depends on NO concentrations and cell types, this highly reactive free radical can activate the p38 MAPK signal cascade. Recent studies have suggested that thioredoxin may play an important role as an effector for some of these functions. Thioredoxin is a major redox protein for many enzymes/transcription factors and is involved in cellular functions, such as viability, activation, and proliferation. In addition to its redox regulation, thioredoxin binds directly to the apoptosis signal-regulating kinase 1 (ASK1), thus inhibiting the activation of stress-induced MAPK signaling cascades that lead to apoptosis. Furthermore, NO produced from newly induced neuronal NOS was reported to induce expression of thioredoxin and several other genes for preconditioning-induced neuroprotection. Moreover, although exposure of endothelial cells to NO decreases NOS activity, this inhibition was shown to be reversed by thioredoxin. Finally, the correlation of expression of thioredoxin with endothelial NOS activity seems to suggest an important role played by this protein in perinatal changes of pulmonary artery functions. Therefore, thioredoxin may participate in the regulation of NOS activity and be involved in NO functions via multiple mechanisms. PMID- 12076973 TI - A critical assessment of the neurodestructive and neuroprotective effects of nitric oxide. AB - Whether nitric oxide is cytodestructive or cytoprotective is of obvious clinical importance. The debate on this subject in the past decade has generated much "heat and light". This paper focuses on the actions of NO on the nervous system and reexamines the controversial issue and the contribution of the authors and their colleagues in the light of recent findings. We also report new findings, critically assesses previous experimental data, and share perspectives on this important subject. PMID- 12076974 TI - Hypothesis: proteasomal dysfunction: a primary event in neurogeneration that leads to nitrative and oxidative stress and subsequent cell death. AB - It is proposed that a primary mechanism leading to neuronal cell death in common neurodegenerative diseases is interference with proteasome function. This can involve genetic defects, direct inactivation of the proteasome (e.g., by reactive oxygen species), or overloading with proteins. The latter can be caused by excessive production of normal proteins or by the formation of poorly degradable proteins as a result of genetic mutations, faulty posttranslational modification, or protein modification by reactive oxygen or nitrogen species. Blockage of the proteasome leads to increased oxidative and nitrative stress, the latter apparently due to upregulation of nitric oxide synthase. Thus, agents that increase proteasome function may be generally neuroprotective, as may be NOS inhibitors. Proteasome inhibitors should be used with caution as therapeutic agents. PMID- 12076975 TI - A chemical perspective on the interplay between NO, reactive oxygen species, and reactive nitrogen oxide species. AB - Nitric oxide (nitrogen monoxide, NO) plays a veritable cornucopia of regulatory roles in normal physiology. In contrast, NO has also been implicated in the etiology and sequela of numerous neurodegenerative diseases that involve reactive oxygen species (ROS) and nitrogen oxide species (RNOS). In this setting, NO is often viewed solely as pathogenic; however, the chemistry of NO can also be a significant factor in lessening injury mediated by both ROS and RNOS. The relationship between NO and oxidation, nitrosation, and nitration reactions is summarized. The salient factors that determine whether NO promotes, abates, or interconnects these chemistries are emphasized. From this perspective of NO chemistry, the type, magnitude, location, and duration of either ROS or RNOS reactions may be predicted. PMID- 12076976 TI - Nitric oxide and superoxide, a deadly cocktail. AB - Free radicals and oxidants may have contrasting effects on cells. Nitric oxide is a cellular messenger that acts by several mechanisms, including activation of soluble guanylate cyclase, nitrosylation of thiols, and formation of peroxynitrite. The action of nitric oxide depends on oxidative conditions in the cell. In motor neurons, nitric oxide enhances brain-derived neurotrophic factor effects by stimulation of cGMP production. In the absence of trophic factors, nitric oxide induces motor neuron apoptosis by reacting with superoxide to form peroxynitrite. The mechanism of induction of motor neuron apoptosis by the formation of peroxynitrite is more complicated than simply the formation of a strong oxidant and it also involves decreased formation of cGMP. PMID- 12076977 TI - Apolipoprotein-E allele-specific regulation of nitric oxide production. AB - Cognitive decline and dementia are key features of Alzheimer's disease (AD) that result from failure of neuronal function. Affected neurons demonstrate indices of nitrosative stress resulting from changes in nitric oxide (NO) mediated redox balance. Neurofibrillary tangles, a characteristic neuropathologic feature of AD, and dysfunctional neurons frequently display 3-nitrotyrosine or other markers of nitrosative stress and immunoreactive nitric oxide synthase (NOS), suggesting that NOS-containing neurons are affected in AD. Our previous studies showed that apolipoprotein E treatment of macrophages increased NO production. Using transgenic mouse models expressing human apoE2, apoE3, or apoE4 protein isoforms and no mouse apoE, we now report an isoform specific difference in microglial NO production. Mice expressing the apoE4 protein isoform have a greater NO production than mice expressing the apoE3 protein isoform. The supply of arginine, the sole substrate for NOS, is dependent on cationic amino acid transporters (CATs) that also demonstrate a similar pattern of apoE isoform dependency. Although arginine transport is greater in APOE4 microglia, this effect is not limited to tissue macrophages. Cortical neurons in primary culture from APOE4 transgenic mice exhibit a similar increase in arginine uptake over neurons cultured from APOE3 mice. The inappropriate levels of arginine transport and of NO in the presence of the APOE4 compared to the APOE3 gene and its products are likely to have significant impact in the CNS. PMID- 12076980 TI - Possible roles of nitric oxide in the physiology and pathophysiology of the mammalian cochlea. AB - Nitric oxide (NO) has been implicated as a mediator of vasodilation and neurotransmission in the mammalian cochlea. This is demonstrated by the presence of nitric oxide synthase (NOS) and nitric oxide (NO) in the blood vessels and the organ of Corti in the cochlea. It is not certain if the neurons in the spiral ganglion produce NO since no fluorescent signal could be detected by 4,5 diaminofluorescein diacetate (DAF-2DA), a fluorescent indicator of NO. To determine if NO/peroxynitrite plays any role in neurodestruction observed in ischemic cochlea of the guinea pig, the effects of NO donors, such as S nitrosocysteine (S-NC) and nitroglycerine (NTG); peroxynitrite generators, such as 3-morpholinosydnonimine (SIN-1); peroxynitrite inhibitors, such as superoxide dismutase plus catalase (SOD/Cat); and NOS inhibitors, such as N(G)-nitro-L arginine methyl ether (L-NAME) were tested on normal and ischemic cochleae. The level of NO in the cochlea after 20 to 120 minutes of ischemia was indicated by measurement of nitrites/nitrates in the perilymph. The evidence gathered from these experiments indicates that NO or peroxynitrite is not necessarily destructive to auditory hair cells, and in fact, exogenous NO may protect neural structures in the cochlea from damage under ischemic conditions. PMID- 12076979 TI - A reassessment of the peroxynitrite scavenging activity of uric acid. AB - Peroxynitrite is implicated in numerous human diseases. Hence, there is considerable interest in potential therapeutic peroxynitrite scavengers. It has been claimed that uric acid is a powerful peroxynitrite scavenger. We previously observed that uric acid is a powerful inhibitor of tyrosine nitration induced by peroxynitrite, but fails to prevent alpha(1)-antiproteinase (alpha(1)-AP) inactivation induced by peroxynitrite. However, the reactivity of peroxynitrite is significantly modified by bicarbonate and this has not been considered in evaluating the scavenging activity of uric acid and other endogenous antioxidant compounds. In the presence of bicarbonate (25 mM), the ability of uric acid, ascorbate, Trolox, and GSH to inhibit peroxynitrite-mediated tyrosine and guanine nitration is decreased. Protection against peroxynitrite-mediated alpha(1)-AP inactivation is also decreased by ascorbate, Trolox, and GSH, but it is enhanced by uric acid. Bicarbonate also inhibits the ability of these compounds to prevent peroxynitrite-mediated ABTS radical cation formation. However, the abilities of these antioxidants to prevent peroxynitrite-mediated bleaching of pyrogallol red are enhanced by bicarbonate. These results show that physiologic concentrations of bicarbonate substantially modify the ability of uric acid to prevent peroxynitrite-mediated reactions. This study highlights the need to use several different assays in the presence of physiologically relevant concentrations of bicarbonate when assessing compounds for peroxynitrite scavenging, in order to avoid misleading results. PMID- 12076978 TI - The association between neuronal nitric oxide synthase and neuronal sensitivity in the brain after brain injury. AB - Injury to the central nervous system is the leading cause of disability in the United States. Neuronal death is one of the causes of disability. Among patients who survive this type of injury, various degrees of recovery in brain function are observed. The molecular basis of functional recovery is poorly understood. Clinical observations and research using experimental injury models have implicated several metabolites in the cascade of events that lead to neuronal degeneration. The levels of intracellular ATP (energy source) and pH are decreased, whereas levels of extracellular glutamate, intracellular calcium ions, and oxidative damage to RNA/DNA, protein, and lipid are increased. These initiating events can be associated with energy failure and mitochondrial dysfunction, resulting in functional or structural brain damage. The injured brain is known to express immediate early genes. Recent studies show that reactive oxygen species (ROS) cause lesions in genes from which mRNA is transcribed as part of the endogenous neuroprotective response. Although degenerating proteins and lipids may contribute to necrosis significantly after severe injury, abnormalities in genetic material, if not repaired, disturb cellular function at every level by affecting replication, transcription, and translation. These lesions include abnormal nucleic acids, known as oxidative lesions of DNA (ODLs) or of RNA (ORLs). In this review, we focus on our current understanding of the various effects of neuronal nitric oxide synthase on the formation of modified bases in DNA and RNA that are induced in the brain after injury, and how ODLs and ORLs affect cell function. PMID- 12076981 TI - Nitric oxide in liver diseases: friend, foe, or just passerby? AB - Research on the free radical gas, nitric oxide (NO), during the past twenty years is one of the most rapid growing areas in biology. NO seems to play a part in almost every organ and tissue. However, there is considerable controversy and confusion in understanding its role. The liver is one organ that is clearly influenced by NO. Acute versus chronic exposure to NO has been associated with distinct patterns of liver disease. In this paper we review and discuss the involvement of NO in various liver diseases collated from observations by various researchers. Overall, the important factors in determining the beneficial versus harmful effects of NO are the amount, duration, and site of NO production. A low dose of NO serves to maximize blood perfusion, prevent platelet aggregation and thrombosis, and neutralize toxic oxygen radicals in the liver during acute sepsis and reperfusion events. NO also demonstrates antimicrobial and antiapoptosis properties during acute hepatitis infection and other inflammatory processes. However, in the setting of chronic liver inflammation, when a large sustained amount of NO is present, NO might become genotoxic and lead to the development of liver cancer. Additionally, during prolonged ischemia, high levels of NO may have cytotoxic effects leading to severe liver injury. In view of the various possible roles that NO plays, the pharmacologic modulation of NO synthesis is promising in the future treatment of liver diseases, especially with the emergence of selective NO synthase inhibitors and cell-specific NO donors. PMID- 12076982 TI - Nitric oxide (NO) and convulsions induced by pentylenetetrazol. AB - Data about the role of nitric oxide (NO) in epileptogenesis are contradictory. It is found to exert both proconvulsant and anticonvulsant effects. In an attempt to elucidate the role of NO in seizures, male Wistar rats were treated intraperitoneally by pentylenetetrazol (PTZ) (60, 80, and 100 mg/kg) and by a nitric oxide synthase antagonist, N-omega-nitro-L-arginine-methyl-ester (L-NAME) (10, 40, and 70 mg/kg), applied before PTZ. The time to onset and incidence of forelimb dystonia (FLD), generalized clonic convulsions (GCC), clonic-tonic convulsions (CTC), and mortality were recorded. The most successful convulsive response and mortality prevention were found in PTZ (80 mg/kg)-treated groups, where L-NAME (70 mg/kg) decreased the incidence by 29, 50, 67 (p = 0.052), and 50%, respectively, and significantly prolonged the time to onset, except that for mortality. Unexpectedly, L-NAME (40 mg/kg) increased incidence of GCC and mortality by 16%, similar to L-NAME (10 mg/kg) in PTZ (60 mg/kg)-treated groups, where GCC, CTC, and mortality increased by 14, 14, and 28%, respectively. Convulsive latency was prolonged in some PTZ (100 mg/kg) + L-NAME (40 and 70 mg/kg)-treated groups. In the experimental model and protocol used, it is concluded that (1) the effects of NO are L-NAME- and PTZ-dose dependent; (2) clonic-tonic convulsions are more strongly influenced by NO than limbic, probably because of PTZ limbic structure overstimulation; (3) L-NAME decreases the incidence of CTC and prolongs FLD, GCC, and CTC times to onset, indicating that NO acts as a proconvulsant; and (3) increased GCC, CTC, and mortality that suggests an anticonvulsant effect of NO needs further investigation. PMID- 12076983 TI - Serum-dependence of LPS-induced neurotoxicity in rat cortical neurons. AB - Previous studies have shown that the bacterial endotoxin, lipopolysaccharide (LPS), is neurotoxic both in vitro and in vivo. The rate of binding of LPS to a target cell is greatly enhanced by serum in general and by LPS binding protein (LBP) in particular. The purpose of the study described in this paper was to determine if microglia activation and LPS-induced neurotoxicity is serum or LBP dependent. A murine microglial cell line, BV2, was used to assess the serum dependence of nitric oxide production and tumor necrosis factor a release in microglia. Embryonic rat cortical neuron/glia mixed cultures were used to determine the serum dependence of LPS-induced neurotoxicity. Our results from both cell culture systems show that LPS-induced inflammatory responses are serum dependent at lower doses of LPS and progressively become serum independent above 10 ng/ml. Purified human recombinant LBP reconstitutes the lost LPS-induced inflammatory responses in primary and immortalized cell cultures treated with heat-denatured serum and appears to account for the serum dependence. These data suggest that the cell surface signaling receptor for LPS at the low and high concentrations are likely to differ, consistent with the existence of a variety of LPS receptors. PMID- 12076984 TI - Role of nitric oxide in inflammation-mediated neurodegeneration. AB - Increasing evidence has suggested that inflammation in the brain is closely associated with the pathogenesis of several degenerative neurologic disorders, including Parkinson's disease, Alzheimer's diseases, multiple sclerosis, amyotrophic lateral sclerosis, and AIDS dementia. The hallmark of brain inflammation is the activation of glial cells, especially that of microglia that produce a variety of proinflammatory and neurotoxic factors, including cytokines, fatty acid metabolites, free radicals--such as nitric oxide (NO) and superoxide. Excessive production of NO, as a consequence of nitric oxide synthase induction in activated glia, has been attributed to participate in neurodegeneration. Using primary mixed neuron-glia cultures and glia-enriched cultures prepared from embryonic rodent brain tissues, we have systemically studied the relationship between the production of NO and neurodegeneration in response to stimulation by the inflammagen lipopolysaccharide. This review summarizes our recent findings on the kinetics of NO generation, the relative contribution of microglia and astrocytes to NO accumulation, the relationship between NO production and neurodegeneration, and points of intervention along the pathways associated with NO generation to achieve neuroprotection. We also describe our results relating to the effect of several opioid-related agents on microglial activation and neuroprotection. Among these agents, the opioid receptor antagonist naloxone, especially its non-opioid enantiomer (+)-naloxone, promises to be of potential therapeutic value for the treatment of inflammation-related diseases. PMID- 12076986 TI - Go6976 protects mesencephalic neurons from lipopolysaccharide-elicited death by inhibiting p38 MAP kinase phosphorylation. AB - Glial activation is associated with inflammation-related neuron degeneration in the brain. A variety of protein kinases are assumed to contribute to the expression of inflammation-related products, such as nitric oxide (NO) and proinflammatory cytokines, however, the mechanisms of glial activation and glia mediated neurotoxicity remain unclear. We found that the indolocarbazole, Go6976, originally known as a selective protein kinase C (PKC) inhibitor, protects neurons from glia-mediated damage and suppresses lipopolysaccharide (LPS)-induced microglial production of inflammatory factors. The purpose of the study we report here was to determine the mechanism underlying the neuroprotective effect of Go6976 in mesencephalic neuron/glia cultures. Go6976 suppressed LPS-induced neurotoxicity in mesencephalic neuron/glia cultures and the protective effect of Go6976 paralleled the suppression of p38 mitogen activated protein kinase (MAPK) activation and inhibition of NO production. Go6976 did not directly inhibit the activity of p38 MAPK; rather, the inhibitor suppressed the phosphorylation of p38 MAPK, suggesting that the target of Go6976 is a signaling event upstream of p38 MAPK. Although Go6976 was originally known to be a selective PKC inhibitor, the neuroprotection was not mediated through its reputed effects on PKC activity. This paper demonstrates that the neuroprotective effect of Go6976 against LPS induced damage is mediated through the inhibition of proinflammatory factors, such as NO from microglia, by inhibiting the phosphorylation of p38 MAPK. PMID- 12076985 TI - p38 MAP kinase is involved in lipopolysaccharide-induced dopaminergic neuronal cell death in rat mesencephalic neuron-glia cultures. AB - Immune stimulants, such as the bacterial endotoxin, lipopolysaccharide (LPS), the human immunodeficiency virus-1 coat protein gp120, or beta-amyloid peptides, lead to glial activation and production of various immune mediators, such as nitric oxide (NO) and proinflammatory cytokines in the brain. These mediators appear to contribute to neuronal cell death in neurodegenerative diseases. However, the signaling pathways, which mediate the neurotoxic effect by the endotoxin, are not understood. The purpose of this study was to determine the role of mitogen activated protein kinase (MAPK) in LPS-induced neurodegeneration using mesencephalic dopaminergic neuron/glia cultures. We have found that the p38 MAPK is important in LPS-induced death of mesencephalic neurons in rat neuron-glia mixed cultures. Upon treatment with 10 ng/ml LPS, the number of dopaminergic neurons decreased by 80% within 48 h, preceded by a significant production of NO by glia. Neuroprotection by selective inhibition of p38 MAPK activity paralleled a decrease in LPS-induced inducible nitric oxide synthase (iNOS) expression. These events were significantly reduced by the selective p38 MAPK inhibitor, SB202190, but not by the inactive analogue SB202474. Inhibition of iNOS activity and NO production by treatment with GW274150 was also neuroprotective. Although the p38 MAPK inhibitor afforded significant neuroprotection from LPS toxicity in the neuron-glia mixed culture, it failed to protect dopaminergic neurons from 6 hydroxy-dopamine-induced toxicity, which acts directly on dopaminergic neurons by inducing hydroxyl radical formation from the mitochondria. The results suggest that p38 MAPK in glia plays a significant role in the LPS-induced death of mesencephalic neurons through induction of nitric oxide synthase and resulting NO production. PMID- 12076987 TI - Nitric oxide-releasing drugs: a novel class of effective and safe therapeutic agents. AB - Nitric oxide (NO) deficiency has been implicated in many pathologic processes, thus providing a solid biological basis for the use of NO replacement therapy. Exogenous NO sources constitute a powerful way to supplement NO when the body cannot generate sufficient NO for normal biological functions. This theory has opened up the possibility of designing new drugs that are capable of delivering NO into tissues and the bloodstream in a sustained and controlled manner. This objective has been achieved by grafting an organic nitrate structure onto existing drugs through chemical spacers, such as aliphatic, aromatic, or a heterocyclic chain. The approach has led to the synthesis of several new chemical entities whose pharmacologic profile challenges the parent drug, not only on the basis of new properties, but also with respect to a better safety profile. In this article, a specific class of NO donors is reviewed, the nitric oxide releasing non-steroidal antiinflammatory drugs, NO-NSAIDs. Recently discovered compounds, whose action depends on the combined properties of both the known drug and NO release, are illustrated. Two examples are described in detail: (1) nitric oxide-releasing aspirin, which has demonstrable innovative properties for treatment of vascular disorders and cancer; (2) nitro-derivatives of flurbiprofen that have shown encouraging results in models of Alzheimer's disease. PMID- 12076988 TI - Ginseng, sex behavior, and nitric oxide. AB - In Asia, ginseng is commonly included in herbals used for the treatment of sexual dysfunction. Recent studies in laboratory animals have shown that both Asian and American forms of ginseng enhance libido and copulatory performance. These effects of ginseng may not be due to changes in hormone secretion, but to direct effects of ginseng, or its ginsenoside components, on the central nervous system and gonadal tissues. Indeed, there is good evidence that ginsenosides can facilitate penile erection by directly inducing the vasodilatation and relaxation of penile corpus cavernosum. Moreover, the effects of ginseng on the corpus cavernosum appear to be mediated by the release and/or modification of release of nitric oxide from endothelial cells and perivascular nerves. Treatment with American ginseng also affects the central nervous system and has been shown to significantly alter the activity of hypothalamic catecholamines involved in the facilitation of copulatory behavior and hormone secretion. Recent findings that ginseng treatment decreased prolactin secretion also suggested a direct nitric oxide-mediated effect of ginseng at the level of the anterior pituitary. Thus, animal studies lend growing support for the use of ginseng in the treatment of sexual dysfunction and provide increasing evidence for a role of nitric oxide in the mechanism of ginsenoside action. PMID- 12076989 TI - The clinical safety of viagra. AB - Erectile dysfunction (ED) in men is amenable to correction with Viagra in a majority of patients. The accumulated experience of prescribing Viagra across the broad continuum of men suffering from ED is sufficient for a meaningful assessment of the safety of Viagra in clinical practice. The use of Viagra necessitates caution in cardiac failure and when used within six months of acute myocardial infarction and stroke. It is inadvisable in patients with unstable angina pectoris. The co-administration of Viagra with organic nitrates, for example, glyceryl trinitrate or isosorbide dinitrate, is unsafe. The relative contraindications to Viagra in cardiovascular disease are uncontrolled hypertension and impaired cardiac reserve. With respect to interactions with other drugs, the potential influence on the metabolism of Viagra by medications that affect the cytochrome-P-450 system does not translate into clinical effects. The vasodilatory properties of sildenafil citrate are largely responsible for unwanted effects. The most common side effects are headache, flushing (due to vasodilation), and dyspepsia (due to relaxation of the smooth muscle of the gastroesophageal sphincter with reflux). In the recommended single-dose range (25 100 mg), the use of Viagra for erectile dysfunction, in the absence of contraindications, is extremely safe provided the drug is taken under proper conditions. PMID- 12076990 TI - Nitric oxide: an antioxidant and neuroprotector. AB - Indirect evidence, including neuroprotection against 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP)-neurotoxicity by nitric oxide synthase (NOS) inhibitors and resistance of transgenic animals deficient in NOS, is controversial. We have reviewed evidence in favor of oxidative stress during the development of MPTP neurotoxicity and the influence of antioxidants, including nitric oxide (NO) and NO donors, on MPTP-induced dopaminergic neurotoxicity. Systemic administration of MPTP causes dose-dependent generation of hydroxyl radicals (OH) in vivo in the striatum in mice; OH scavengers protect dopaminergic neurons from this insult. On the other hand the role of NO in MPTP-neurotoxicity is controversial. Hitherto, no direct evidence for the involvement of NO in MPTP neurotoxicity has been available. MPTP does not affect inducible-NOS mRNA level or its expression in SN or the striatum. Nitroglycerine, a NO donor, can attenuate MPTP-induced dopamine depletion in the striatum by virtue of its OH scavenging action. Several other NO donors have also been shown to scavenge the OH generated, following Fenton chemistry in vitro, and to protect against in vivo dopaminergic neurotoxicity by small mass iron complex formation. This evidence suggests that NO renders protection against MPTP-induced OH-mediated nigrostriatal lesions, acting as an antioxidant. PMID- 12076991 TI - Nitric oxide triggers classic ischemic preconditioning. AB - The role of NO in the classic ischemic preconditioning phenomenon of the myocardium is not well defined, and was investigated by using the isolated perfused rat heart as a model. Hearts were preconditioned with 3 x 5 minute ischemia in the presence and absence of the NOS inhibitors L-NAME (50 microM) and L-NNA (50 microM), and the guanylyl cyclase inhibitor ODQ (20 microM). These inhibitors significantly attenuated the protective effect of preconditioning against 25-min global ischemia (as measured by functional recovery), specifically if administered during the triggering phase. Cyclic infusions (3 x 5 min) of the NO-donors SNAP (50 microM) and SNP (100 microM) elicited protection against both 25-min global or low-flow ischemia. Hearts preconditioned with NO donors displayed significantly superior functional reserve, if stimulated with adrenaline, compared to hearts preconditioned with ischemia. Although the NO donors SNAP and SNP both activated p38 MAPK during the preconditioning protocol, protection was accompanied by significantly decreased p38 MAPK activity during sustained ischemia, as was the case in ischemic preconditioning. We conclude that (1) NO is a trigger for classic preconditioning, (2) cGMP generation plays an important role in its protection, (3) attenuation of p38 MAPK during sustained ischemia accompanies NO preconditioning and may mediate cardiac protection, and (4) preconditioning with NO may be more advantageous than using ischemia. PMID- 12076992 TI - Mechanisms of hepatoprotection by nitric oxide. AB - Nitric oxide (NO) exerts numerous antiapoptotic effects on hepatocytes in settings of inflammation and tissue damage. These actions of NO are modulated by a variety of mechanisms under both physiologic and pathologic conditions. Nitric oxide inhibits cell death or apoptosis by modulation of heat shock proteins, S nitrosylation of caspases at their catalytic site cysteine residue, triggering of the cGMP pathway, and prevention of mitochondrial dysfunction. Our preliminary studies also suggest that NO can modulate apoptosis-related genes in a manner consistent with an antiapoptotic effect. This review focuses on these molecular mechanisms of cytoprotection by NO. PMID- 12076994 TI - Devic's disease: bridging the gap between laboratory and clinic. PMID- 12076993 TI - Protective effects of glial cell line-derived neurotrophic factor in ischemic brain injury. AB - Glial cell line-derived neurotrophic factor (GDNF), a member of the transforming growth factor-beta (TGF-beta) superfamily, has been shown to have trophic activity on dopaminergic neurons. Recent studies indicate that GDNF can protect the cerebral hemispheres from damage induced by middle cerebral arterial ligation. We found that such neuroprotective effects are mediated through specific GDNF receptor alpha-1 (GFRalpha1). Animals with a deficiency in GFRalpha 1 have less GDNF-induced neuroprotection. Ischemia also enhances nitric oxide synthase (NOS) activity, which can be attenuated by GDNF. These.data suggest that GDNF can protect against ischemic injury through a GFRalpha-1/NOS mechanism. We also found that the receptor for GDNF, GFRalpha1, and its signaling moiety c-Ret were upregulated, starting immediately after ischemia. This upregulation suggests that activation of an endogenous neuroprotective mechanism occurs so that responsiveness of GDNF can be enhanced at very early stages during ischemia. PMID- 12076995 TI - The neuropathology of primary mood disorder. AB - The biological mechanisms proposed to underlie primary mood disorder do not usually include a neuropathological component. However, a significant MRI literature attests to structural abnormalities in regions and has encouraged neuropathological investigations from which candidate histological correlates have begun to emerge. In particular, there are several reports of cytoarchitectural alterations in anterior cingulate and prefrontal cortices, characterized by a decrease in the number or density of glia. Reductions in the size and density of some neuronal populations have also been described, accompanied by alterations in indices of synaptic terminals and dendrites. This form of pathology putatively reflects aberrant neurodevelopment or impaired cellular plasticity. A separate pathological process is suggested by the excess of subcortical focal lesions seen on MRI, especially in elderly patients; these probably reflect white matter damage of vascular origin. Both types of pathology have been observed, to a greater or lesser extent, in unipolar as well as bipolar mood disorders. None of the findings appear attributable to treatment with antidepressants, mood stabilizers or electroconvulsive therapy (ECT). However, all findings remain preliminary due to a lack of unequivocal replication and the failure to control fully for other potential confounders and co-morbid conditions. There are also basic questions to be answered concerning the clinical correlates, magnitude, progression and heterogeneity of the pathology. Nevertheless, it must now be considered likely that changes in brain structure, both macroscopic and microscopic, are a feature of primary mood disorder, a fact to be taken into account when interpreting functional imaging, neuropsychological and neurochemical data. The neuropathology is postulated to contribute to the pathophysiology and dysfunction of the neural circuits which regulate mood and its associated cognitions, behaviours and somatic symptoms. PMID- 12076996 TI - A role for humoral mechanisms in the pathogenesis of Devic's neuromyelitis optica. AB - Devic's disease [neuromyelitis optica (NMO)] is an idiopathic inflammatory demyelinating disease of the CNS, characterized by attacks of optic neuritis and myelitis. The mechanisms that result in selective localization of inflammatory demyelinating lesions to the optic nerves and spinal cord are unknown. Serological and clinical evidence of B cell autoimmunity has been observed in a high proportion of patients with NMO. The purpose of this study was to investigate the importance of humoral mechanisms, including complement activation, in producing the necrotizing demyelination seen in the spinal cord and optic nerves. Eighty-two lesions were examined from nine autopsy cases of clinically confirmed Devic's disease. Demyelinating activity in the lesions was immunocytochemically classified as early active (21 lesions), late active (18 lesions), inactive (35 lesions) or remyelinating (eight lesions) by examining the antigenic profile of myelin degradation products within macrophages. The pathology of the lesions was analysed using a broad spectrum of immunological and neurobiological markers, and lesions were defined on the basis of myelin protein loss, the geography and extension of plaques, the patterns of oligodendrocyte destruction and the immunopathological evidence of complement activation. The pathology was identical in all nine patients. Extensive demyelination was present across multiple spinal cord levels, associated with cavitation, necrosis and acute axonal pathology (spheroids), in both grey and white matter. There was a pronounced loss of oligodendrocytes within the lesions. The inflammatory infiltrates in active lesions were characterized by extensive macrophage infiltration associated with large numbers of perivascular granulocytes and eosinophils and rare CD3(+) and CD8(+) T cells. There was a pronounced perivascular deposition of immunoglobulins (mainly IgM) and complement C9neo antigen in active lesions associated with prominent vascular fibrosis and hyalinization in both active and inactive lesions. The extent of complement activation, eosinophilic infiltration and vascular fibrosis observed in the Devic NMO cases is more prominent compared with that in prototypic multiple sclerosis, and supports a role for humoral immunity in the pathogenesis of NMO. Based on this study, future therapeutic strategies designed to limit the deleterious effects of complement activation, eosinophil degranulation and neutrophil/macrophage/microglial activation are worthy of further investigation. PMID- 12076998 TI - A multigenerational family with multiple sclerosis. AB - We report a family with 15 individuals affected with multiple sclerosis present in three and possibly four generations. The segregation of multiple sclerosis within this pedigree is consistent with an autosomal dominant mode of inheritance with reduced penetrance. The clinical characteristics of the affected individuals are indistinguishable from those seen in sporadic multiple sclerosis with respect to sex ratio, age at onset, onset symptom, MRI and clinical course. Eleven of 14 cases (78.6%) were positive for the known multiple sclerosis-associated major histocompatibility complex (MHC) Class II HLA DRB1*15 allele. Parametric linkage analysis gave a non-significant LOD score of 0.31 (theta; = 0.33) for the DRB1 gene. However, among 11 affected children with at least one DRB1*15 bearing parent, all 11 out of 11 received at least one copy of this known susceptibility allele. A transmission disequilibrium test analysis was significant for the DRB1*15 allele within this single family; P = 0.0054. The inheritance pattern in this family suggests the presence of a single major locus responsible for multiple sclerosis susceptibility, with DRB1 acting as an important modifier. This family could be an important resource for the identification of a multiple sclerosis susceptibility gene. PMID- 12076997 TI - Markers for different glial cell responses in multiple sclerosis: clinical and pathological correlations. AB - Disease progression in multiple sclerosis occurs within the interface of glial activation and gliosis. This study aimed to investigate the relationship between biomarkers of different glial cell responses: (i) to disease dynamics and the clinical subtypes of multiple sclerosis; (ii) to disability; and (iii) to cross validate these findings in a post-mortem study. To address the first goal, 51 patients with multiple sclerosis [20 relapsing remitting (RR), 21 secondary progressive (SP) and 10 primary progressive (PP)] and 51 neurological control patients were included. Disability was assessed using the ambulation index (AI), the Expanded Disability Status Scale score (EDSS) and the 9-hole PEG test (9HPT). Patients underwent lumbar puncture within 7 days of clinical assessment. Post mortem brain tissue (12 multiple sclerosis and eight control patients) was classified histologically and adjacent sites were homogenized for protein analysis. S100B, ferritin and glial-fibrillary acidic protein (GFAP) were quantified in CSF and brain-tissue homogenate by ELISA (enzyme-linked immunosorbent assay) techniques developed in-house. There was a significant trend for increasing S100B levels from PP to SP to RR multiple sclerosis (P < 0.05). S100B was significantly higher in RR multiple sclerosis than in control patients (P < 0.01), whilst ferritin levels were significantly higher in SP multiple sclerosis than in control patients (P < 0.01). The S100B : ferritin ratio discriminated patients with RR multiple sclerosis from SP, PP or control patients (P < 0.05, P < 0.01 and P < 0.01, respectively). Multiple sclerosis patients with poor ambulation (AI > or =7) or severe disability (EDSS >6.5) had significantly higher CSF GFAP levels than less disabled multiple sclerosis or control patients (P < 0.01 and P < 0.001, respectively). There was a correlation between GFAP levels and ambulation in SP multiple sclerosis (r = 0.57, P < 0.01), and between S100B level and the 9HPT in PP multiple sclerosis patients (r = -0.85, P < 0.01). The post-mortem study showed significantly higher S100B levels in the acute than in the subacute plaques (P < 0.01), whilst ferritin levels were elevated in all multiple sclerosis lesion stages. Both GFAP and S100B levels were significantly higher in the cortex of multiple sclerosis than in control brain homogenate (P < 0.001 and P < 0.05, respectively). We found that S100B is a good marker for the relapsing phase of the disease (confirmed by post-mortem observation) as opposed to ferritin, which is elevated throughout the entire course. GFAP correlated with disability scales and may therefore be a marker for irreversible damage. The results of this study have broad implications for finding new and sensitive outcome measures for treatment trials that aim to delay the development of disability. They may also be considered in future classifications of multiple sclerosis patients. PMID- 12076999 TI - Nicotinic receptor abnormalities in the cerebellar cortex in autism. AB - Autism is a common developmental disorder associated with structural and inferred neurochemical abnormalities of the brain. Cerebellar abnormalities frequently have been identified, based on neuroimaging or neuropathology. Recently, the cholinergic neurotransmitter system has been implicated on the basis of nicotinic receptor loss in the cerebral cortex. Cerebellar cholinergic activities were therefore investigated in autopsy tissue from a series of autistic individuals. The presynaptic cholinergic enzyme, choline acetyltransferase, together with nicotinic and muscarinic receptor subtypes were compared in the cerebellum from age-matched mentally retarded autistic (eight), normal control (10) and non autistic mentally retarded individuals (11). The nicotinic receptor binding the agonist epibatidine (the high affinity receptor subtype, consisting primarily of alpha3 and alpha4, together with beta2 receptor subunits) was significantly reduced by 40-50% in the granule cell, Purkinje and molecular layers in the autistic compared with the normal group (P < 0.05). There was an opposite increase (3-fold) in the nicotinic receptor binding alpha-bungarotoxin (to the alpha7 subunit) which reached significance in the granule cell layer (P < 0.05). These receptor changes were paralleled by a significant reduction (P < 0.05) and non-significant increase, respectively, of alpha4 and alpha7 receptor subunit immunoreactivity measured using western blotting. Immunohistochemically loss of alpha(4 )reactivity was apparent from Purkinje and the other cell layers, with increased alpha7 reactivity in the granule cell layer. There were no significant changes in choline acetyltransferase activity, or in muscarinic M1 and M2 receptor subtypes in autism. In the non-autistic mentally retarded group, the only significant abnormality was a reduction in epibatidine binding in the granule cell and Purkinje layers. In two autistic cases examined histologically, Purkinje cell loss was observed in multiple lobules throughout the vermis and hemispheres. This was more severe in one case with epilepsy, which also showed vermis folial malformation. The case with less severe Purkinje cell loss also showed cerebellar white matter thinning and demyelination. These findings indicate a loss of the cerebellar nicotinic alpha4 receptor subunit in autism which may relate to the loss of Purkinje cells, and a compensatory increase in the alpha7 subunit. It remains to be determined how these receptor abnormalities are involved in neurodevelopment in autism and what is the relationship to mental function. Since nicotinic receptor agonists enhance attentional function and also induce an elevation in the high affinity receptor, nicotinic therapy in autism may be worth considering. PMID- 12077000 TI - Stimulation of the greater occipital nerve induces increased central excitability of dural afferent input. AB - Patients with primary headaches often report pain that involves not only the front of the head, innervated by the first (ophthalmic) division of the trigeminal nerve, but also the back of the head, innervated by the greater occipital nerve (GON) that is a branch of the C(2) spinal root. The aim of this work was to study the physiology of trigeminocervical input in a model of cranial nociception by describing a population of nociceptive neurones that receive convergent input from the supratentorial dura and the GON. Rats were anaesthetized with pentobarbital, paralysed and ventilated. The parietal dura above the middle meningeal artery was stimulated through a closed cranial window. The GON was exposed in the neck and stimulated. Recordings were made from 67 neurones (52 wide dynamic range neurones/15 nociceptive-specific-neurones) in the C(2) spinal dorsal horn, which responded to stimulation of the dura and the GON. Most neurones showed receptive fields corresponding to the first trigeminal division as well as input from C(2) skin and muscle. Neurones were recorded in superficial and deep layers of the dorsal horn. All neurones tested received input from the ipsilateral and contralateral GON (n = 5). The responses to dural stimulation were analysed before and after stimulation of the GON. Supramaximal electrical stimulation of the GON (20 s to 5 min) enhanced afferent dural input in 8/20 neurones. Application of the C-fibre activator mustard oil (MO) to the cutaneous receptive field or suboccipital muscles innervated by the GON induced an increased excitability of dural responses in 8/12 and 9/10 neurones, respectively. Stimulation of muscle afferents had a significant longer facilitatory effect than cutaneous stimulation (P < 0.05). These results show that a considerable population of neurones show convergent input from both dura as well as cervical cutaneous and muscle territories, which supports the view of a functional continuum between the caudal trigeminal nucleus and upper cervical segments involved in cranial nociception. The facilitatory effect of GON stimulation on dural stimulation suggests a central mechanism at the second order neurone level. This mechanism may be important in pain referral from cervical structures to the head and therefore have implications for most forms of primary headache. PMID- 12077001 TI - Change of chloride ion channel conductance is an early event of slow-to-fast fibre type transition during unloading-induced muscle disuse. AB - Disuse of postural slow-twitch muscles, as it occurs in hypogravity, induces a slow-to-fast myofibre type transition. Nothing is known about the effects of weightlessness on the resting membrane chloride conductance (gCl), which controls sarcolemma excitability and influences fibre type transition during development and adult life. Using the current-clamp method, we observed that rat hindlimb unloading (HU) for 1-3 weeks increased gCl in fibres of the slow-twitch soleus (Sol) muscle toward values found in fast muscle. Northern blot analysis suggested that this effect resulted from an increased ClC-1 chloride channel mRNA level. In the meantime, a 4-fold increase in fibres expressing fast isoforms of the myosin heavy chain (MHC) was observed by immunostaining of muscle sections. Also, Sol muscle function evolved toward a fast phenotype during HU, as demonstrated by the positive shift of the threshold potential for contraction. After 3-days HU, Sol muscle immunostaining and RT-PCR experiments revealed no change in MHC protein and mRNA expression, whereas the gCl was already maximally increased, due to a pharmacologically probed, increased activity of ClC-1 channels. Thus the increase in gCl is an early event in Sol muscle experiencing unloading, suggesting that gCl may play a role in muscle adaptation to modified use. Pharmacological modulation of ClC-1 channels may help to prevent disuse-induced muscle impairment. PMID- 12077002 TI - Mitochondrial dysfunction in a cell culture model of familial amyotrophic lateral sclerosis. AB - The molecular mechanisms by which mutations in the gene for Cu/Zn superoxide dismutase (SOD1) lead to the selective death of motor neurones in familial amyotrophic lateral sclerosis (FALS) remain incompletely understood. Previous evidence has indicated that mitochondrial abnormalities may develop during motor neurone injury, but several important questions remain unanswered. We have developed a cell culture model of FALS in which a motor neurone cell line (NSC34) has been stably transfected to express normal or mutant human SOD1 at levels approximating to those seen in the human disease. The aims of the study were to: (i) investigate whether morphological mitochondrial abnormalities occur at expression levels of mutant SOD1 close to physiological levels; and (ii) determine whether the presence of mutant SOD1 causes abnormalities of mitochondrial respiratory chain function and changes in cellular bioenergetic parameters in motor neuronal cells. Using this cellular model, we demonstrate that the presence of mutant SOD1 results in the development of abnormally swollen and pale staining mitochondria. These morphological changes are accompanied by biochemical abnormalities with specific decreases in the activities of complexes II and IV of the mitochondrial electron transfer chain. These same complexes are inhibited when control NSC34 cells are subjected to oxidative stress induced by serum withdrawal. The decrease in respiratory chain complex activity in the presence of mutant SOD1 was not accompanied by decreased expression of representative proteins present in these complexes. Motor neuronal cells expressing mutant SOD1 showed increased cell death when exposed to oxidative stress by serum withdrawal, whereas the presence of normal human SOD1 exerted a protective effect. Under basal, unstressed culture conditions, no change in the ATP : ADP ratio was observed in the presence of mutant SOD1. However, the mitochondrial changes associated with the presence of mutant SOD1 clearly had adverse cellular bioenergetic consequences as shown by increased cell death in the presence of pharmacological inhibition of the glycolytic pathway. We conclude that one important mechanism by which mutant SOD1 causes motor neurone injury involves inhibition of specific components of the mitochondrial electron transfer chain. Therapeutic measures aimed at protecting mitochondrial respiratory chain function may be useful in SOD1 related familial and possibly other forms of amyotrophic lateral sclerosis. PMID- 12077003 TI - Two populations of neuronal intranuclear inclusions in SCA7 differ in size and promyelocytic leukaemia protein content. AB - Spinocerebellar ataxia type 7 (SCA7) is a hereditary progressive cerebellar ataxia with retinal degeneration associated with an abnormally expanded polyglutamine stretch. Neuronal intranuclear inclusions (NIIs), as in other polyglutamine diseases, are pathological hallmarks of these disorders. NIIs in polyglutamine diseases contain not only the protein with the expanded polyglutamine stretch but also other types of proteins. Several chaperone proteins related to the ubiquitin proteasome pathway, transcription factors and nuclear matrix proteins have been detected in NIIs. The composition of NIIs might reflect the process of NII formation and part of the pathogenesis of these diseases. To investigate how these proteins relate to the pathogenesis of SCA7, we performed immunohistochemical analyses of the composition of NIIs in two cases of SCA7. We demonstrated that there are two types of NIIs in SCA7 that differ in size and immunoreactivity to promyelocytic leukaemia protein (PML), one of the essential components of nuclear bodies (NBs; also called PML oncogenic domains). Small and large NIIs contained ataxin-7, human DnaJ homologue 2 (HDJ-2) and proteasome subunit 19S. In contrast, PML was found only in small NIIs. CREB binding protein (CBP), another component of NBs, was distributed like PML in NIIs. Our results suggest that NIIs are formed by the accumulation of ataxin-7 in NBs, which become enlarged as they recruit related proteins. PMID- 12077004 TI - Cerebellar hemispheric activation ipsilateral to the paretic hand correlates with functional recovery after stroke. AB - An experimental lesion in the primary motor or sensory cortices in monkeys leads to functional reorganization in areas surrounding the lesion or in contralateral homologous regions. In humans, task-dependent brain activation after motor stroke seems to be multifocal and bilateral. Although many active structures are seen after stroke, their roles are unclear. For instance, the uninjured primary motor cortex may play a significant role in recovery or may be associated with mirror movements. Other motor areas, particularly those outside the affected middle cerebral artery distribution, have also been thought to play such a role, including the medial pre-motor areas and both cerebellar hemispheres. The lateral pre-motor areas might also contribute but the demarcation of primary motor and pre-motor cortices is not trivial. It is not known from existing studies how brain activation relates to behavioural change over the time course of recovery. We used functional MRI (fMRI) to study 12 patients longitudinally over the first 6 months of stroke recovery. All subjects had acute stroke causing unilateral arm weakness and had some ability to move the impaired hand within 1 month. Each patient had both motor testing and fMRI during finger and wrist movements at four points during the observed period. Six of these patients showed good motor recovery, whereas the other six did not. The imaging results support a role for the cerebellum in mediating functional recovery from stroke. The data suggest that patients with good recovery have clear changes in the activation of the cerebellar hemisphere opposite the injured corticospinal tract. Patients with poor recovery do not show such changes in cerebellar activation. No other brain region had a significant correlation with recovery. Interestingly, activation in the cerebellum ipsilateral to the injury increases transiently after stroke, independently of the success of recovery. The present work suggests a possible link between cerebellar activation and behavioural recovery from hand weakness from stroke. The underlying mechanism is not known, but it could relate to haemodynamic changes such as diaschisis or to the postulated role of the cerebellum in motor skill learning. PMID- 12077006 TI - Adaptive modification of saccade amplitude in Parkinson's disease. AB - The accuracy of saccades (fast eye movements) is maintained over time and is an adaptive ability usually ascribed to the cerebellum. Adaptation might occur elsewhere in certain tasks, such as in the prefrontal cortex for memory-guided saccades. We hypothesized that adaptation of memory-guided saccades would be impaired in Parkinson's disease, as basal ganglia dysfunction can disrupt the operation of the prefrontal cortex, while adaptation of visually guided saccades would be preserved. Adaptation was induced by consistently yet imperceptibly displacing targets as saccades were made toward them, causing artificial saccadic inaccuracy. Twelve Parkinson's disease subjects (OFF medication) and 12 age matched controls performed 245 visually- and memory-guided horizontal saccades in separate sessions. An infrared eye tracker detected the saccade, during which the target was displaced by 12.5% of the size of the initial jump, either in the same (centrifugal) or the opposite (centripetal) direction. Parkinson's disease subjects made smaller visually guided saccades than did controls [F(1,20) = 9.10, P < 0.01], yet both groups modified saccade size appropriately. Parkinson's disease memory-guided saccades were also smaller than those of controls [F(1,19) = 5.93, P < 0.05]. While controls decreased (by 8.6%) or increased (by 4.1%) the size of these saccades appropriately, Parkinson's disease subjects decreased saccade size in response to both centripetal adaptation (by an excessive 18.3%) and centrifugal adaptation (by 3.5%). Parkinson's disease subjects were less able to modify saccadic size appropriately when the movement size was specified in motor memory: a predilection for excessive hypometria was invoked, regardless of adaptation direction. This indicates that, in certain tasks, saccadic adaptation involves structures other than the cerebellum. PMID- 12077005 TI - Dopamine-dependent changes in the functional connectivity between basal ganglia and cerebral cortex in humans. AB - We test the hypothesis that interaction between the human basal ganglia and cerebral cortex involves activity in multiple functional circuits characterized by their frequency of oscillation, phase characteristics, dopamine dependency and topography. To this end we took recordings from macroelectrodes (MEs) inserted into the subthalamic nucleus (STN) in eight awake patients following functional neurosurgery for Parkinson's disease. An EEG was also recorded, as were the signals from MEs in the globus pallidus interna (GPi) in two of the cases. Coherence between EEG and ME potentials was apparent in three major frequency bands, 2-10 Hz, 10-30 Hz and 70-85 Hz. These rhythmic activities differed in their cortical topography, although coherence was always strongest over the midline. Coherence between EEG and ME potentials in the 70-85 Hz band was only recorded in patients treated with levodopa. Cortical activity phase led that in the basal ganglia in those oscillatory activities with frequencies <30 Hz. In contrast, STN and GPi phase led cortex in the 70-85 Hz band. The temporal differences in the way in which cortical activity led or lagged behind that in STN/GPi were similar, around 20 ms, regardless of the overall direction of information flow and frequency band. We conclude that the basal ganglia may receive multiple cortical inputs at frequencies <30 Hz and, in the presence of dopaminergic activity, produce a high frequency drive back to the cerebral cortex, in particular the supplementary motor area (SMA). PMID- 12077007 TI - Neural systems underlying British Sign Language and audio-visual English processing in native users. AB - In order to understand the evolution of human language, it is necessary to explore the neural systems that support language processing in its many forms. In particular, it is informative to separate those mechanisms that may have evolved for sensory processing (hearing) from those that have evolved to represent events and actions symbolically (language). To what extent are the brain systems that support language processing shaped by auditory experience and to what extent by exposure to language, which may not necessarily be acoustically structured? In this first neuroimaging study of the perception of British Sign Language (BSL), we explored these questions by measuring brain activation using functional MRI in nine hearing and nine congenitally deaf native users of BSL while they performed a BSL sentence-acceptability task. Eight hearing, non-signing subjects performed an analogous task that involved audio-visual English sentences. The data support the argument that there are both modality-independent and modality-dependent language localization patterns in native users. In relation to modality independent patterns, regions activated by both BSL in deaf signers and by spoken English in hearing non-signers included inferior prefrontal regions bilaterally (including Broca's area) and superior temporal regions bilaterally (including Wernicke's area). Lateralization patterns were similar for the two languages. There was no evidence of enhanced right-hemisphere recruitment for BSL processing in comparison with audio-visual English. In relation to modality-specific patterns, audio-visual speech in hearing subjects generated greater activation in the primary and secondary auditory cortices than BSL in deaf signers, whereas BSL generated enhanced activation in the posterior occipito-temporal regions (V5), reflecting the greater movement component of BSL. The influence of hearing status on the recruitment of sign language processing systems was explored by comparing deaf and hearing adults who had BSL as their first language (native signers). Deaf native signers demonstrated greater activation in the left superior temporal gyrus in response to BSL than hearing native signers. This important finding suggests that left- temporal auditory regions may be privileged for processing heard speech even in hearing native signers. However, in the absence of auditory input this region can be recruited for visual processing. PMID- 12077008 TI - Brain anatomy and sensorimotor gating in Asperger's syndrome. AB - Asperger's syndrome (an autistic disorder) is characterized by stereotyped and obsessional behaviours, and pervasive abnormalities in socio-emotional and communicative behaviour. These symptoms lead to social exclusion and a significant healthcare burden; however, their neurobiological basis is poorly understood. There are few studies on brain anatomy of Asperger's syndrome, and no focal anatomical abnormality has been reliably reported from brain imaging studies of autism, although there is increasing evidence for differences in limbic circuits. These brain regions are important in sensorimotor gating, and impaired 'gating' may partly explain the failure of people with autistic disorders to inhibit repetitive thoughts and actions. Thus, we compared brain anatomy and sensorimotor gating in healthy people with Asperger's syndrome and controls. We included 21 adults with Asperger's syndrome and 24 controls. All had normal IQ and were aged 18-49 years. We studied brain anatomy using quantitative MRI, and sensorimotor gating using prepulse inhibition of startle in a subset of 12 individuals with Asperger's syndrome and 14 controls. We found significant age related differences in volume of cerebral hemispheres and caudate nuclei (controls, but not people with Asperger's syndrome, had age-related reductions in volume). Also, people with Asperger's syndrome had significantly less grey matter in fronto-striatal and cerebellar regions than controls, and widespread differences in white matter. Moreover, sensorimotor gating was significantly impaired in Asperger's syndrome. People with Asperger's syndrome most likely have generalized alterations in brain development, but this is associated with significant differences from controls in the anatomy and function of specific brain regions implicated in behaviours characterizing the disorder. We hypothesize that Asperger's syndrome is associated with abnormalities in fronto striatal pathways resulting in defective sensorimotor gating, and consequently characteristic difficulties inhibiting repetitive thoughts, speech and actions. PMID- 12077009 TI - Cortical motor reorganization after a single clinical attack of multiple sclerosis. AB - In order to evaluate whether cortical motor reorganization occurs in the earliest phase of multiple sclerosis, we studied patients after a first clinical attack of hemiparesis. From a consecutive series of 70 patients enrolled in a study of patients with clinically isolated syndrome and serial MRI findings indicative of multiple sclerosis, we retrospectively selected 10 patients with hemiparesis as the onset symptom and no further clinical episode [mean age 32 +/- 9 years, disease duration 24 +/- 14 months, median Expanded Disability Status Score (EDSS) 1.25]. Ten age-matched, healthy subjects served as controls. Each subject was submitted to two functional MRI trials (one per hand) using a 1.5 T magnet during a sequential finger-to-thumb opposition task. Image analysis was performed using SPM99 software. Movements of both the 'affected' and the 'unaffected' hand activated significantly larger areas in patients than in controls in both the contralateral and ipsilateral cortical motor areas. Patients activated a greater number of foci than controls during both the right-hand and the left-hand movement. Most of these foci were located in cortical areas which were less or not at all activated in controls, such as the lateral premotor cortex [Brodmann area (BA) 6], the insula and the inferior parietal lobule (BA 40). Between-group analysis of patients versus controls showed significant (P < 0.001) foci in these areas, principally located in the ipsilateral hemisphere during right-hand movement and in both the cerebral hemispheres during left-hand movement. Time since clinical onset showed a significant positive correlation with the extent of activation in the ipsilateral motor areas (P = 0.006) during the right-hand movement and with the extent of activation in both the ipsilateral (P = 0.02) and contralateral (P = 0.006) motor areas during the left-hand movement. The T(1) lesion load along the motor pathway showed a significant positive correlation (P = 0.007) with the extent of activation in the contralateral motor areas during right-hand movement. Our study shows functional adaptive changes that involve both the symptomatic and asymptomatic hemisphere during a simple motor task in patients who had suffered a single clinical attack of hemiparesis. The extent of these changes increased with the time elapsed since disease onset and the severity of brain damage. PMID- 12077010 TI - Adolescents who were born very preterm have decreased brain volumes. AB - Infants born very preterm have an increased risk of brain injury. Given the great increase in the number of such infants that are surviving, it is important to establish whether any resultant brain abnormalities persist into adolescence and adult life. We therefore examined in vivo whole brain, grey matter, white matter and hippocampal volumes, ventricular size and grey/white matter ratios in a series of adolescents who had been born very preterm, and an age-matched full term control group. Structural MRI was carried out on a cohort of 72 adolescents (mean age 15 years) who were born before 33 weeks, and 48 age-matched full-term controls. Brain measurements were made blind to group affiliation using stereological principles. After controlling for gender and height, the very preterm subjects showed a 6.0% decrease in whole brain volume, and an 11.8% decrease in cortical grey matter volume, as well as a 15.6% decrease in right and a 12.1% decrease in left hippocampal volumes; they also had a 42.0% increase in the size of the lateral ventricles. Therefore, individuals who were born very preterm continue to show noticeable decrements in brain volumes and striking increases in lateral ventricular volume into adolescence. The functional significance of these abnormalities merits further investigation. PMID- 12077011 TI - Neuronal death is enhanced and begins during foetal development in type I spinal muscular atrophy spinal cord. AB - Spinal muscular atrophy (SMA) is an autosomal recessive disorder caused by mutations in the survival motor neurone gene (SMN). The degeneration and loss of the anterior horn cells is the major neuropathological finding in SMA, but the mechanism and timing of this abnormal motor neurone death remain unknown. A quantitative study was carried out comparing neuronal death in controls and SMA foetuses and neonates. Between 12 and 15 weeks of gestational age, a significant increase in nuclear DNA vulnerability, as revealed with the method of in situ end labelling of nuclear DNA fragmentation, was detected in SMA foetuses and was reflected by a decrease in the number of neurones of the anterior horn. Neurones with nuclear DNA vulnerability are no longer detected at the end of the foetal period and the post-natal period. On the other hand, abnormal morphology of motor neurones, mainly early chromatolytic changes, was observed only after birth. Our findings indicate that in type I SMA, the absence or dysfunction of SMN is reflected by an enhanced neuronal death that is already detectable at 12 weeks, the earliest SMA foetal stage analysed. This is associated with a progressive loss of motor neurones towards the neonatal period. Given that a proportion of the remaining SMA motor neurones in the neonatal period appear with pathological findings not detected at earlier stages, it can be hypothesized that type I SMA results in differential age-dependent responses leading to cell death and motor neurone degeneration during development. PMID- 12077012 TI - Reduced oxidative phosphorylation and proton efflux suggest reduced capillary blood supply in skeletal muscle of patients with dermatomyositis and polymyositis: a quantitative 31P-magnetic resonance spectroscopy and MRI study. AB - Quantitative MRI and phosphorus magnetic resonance spectroscopy ((31)P-MRS) were used to investigate skeletal muscle metabolism in vivo in patients with dermatomyositis (DM) and polymyositis (PM) in order to evaluate the role of mitochondrial abnormalities in the pathogenesis and clinical expression of these conditions. Nine patients with DM (mean age +/- SD, 57 +/- 14 years) and five with PM (42 +/- 12 years) and with age at disease onset 53 +/- 16 and 38 +/- 12 years, respectively, were included in the study together with 18 age-matched controls. Post-exercise (31)P-MRS indices of muscle oxidative metabolism were all impaired in DM and PM. In both groups of patients, the phosphocreatine and adenosine diphosphate recovery half-times were almost twice as long as in controls (P < 0.05 for each variable) and the maximum rate of mitochondrial ATP production was half that found in normal subjects (P < 0.001). The rate of proton efflux from muscle fibres was significantly reduced in DM (P < 0.001) and PM (P = 0.02). The impairment of (31)P-MRS recovery indices in DM and PM patients was similar to that found in a group of 10 patients with a primary mitochondrial disorder that showed a normal proton efflux rate. There was no correlation between the MRS-detectable abnormalities and the degree of inflammation or fatty infiltration of the muscle, as measured by MRI. The in vivo findings in DM and PM patients indicate impaired muscle aerobic function, which, considering the reduced proton efflux, is likely to be secondary to an impaired blood supply. Our results suggest that the abnormal mitochondria seen in some muscle biopsies are unlikely to be the primary cause of the oxidative insufficiency in these patients. PMID- 12077013 TI - Neurobehavioural functioning in neonatal intensive care unit graduates in late childhood and early adolescence. AB - The current study examined memory and executive functioning in a sample of children who were hospital ized in a neonatal intensive care unit (NICU) after birth. Thirty-two children born prematurely and/or with medical complications (NICU children) and 25 control children born at term were assessed with the Cambridge Neuropsychological Testing Automated Battery (CANTAB), a multi dimensional computer-based measure of memory and executive functioning. Comparisons between the NICU and control groups on the CANTAB subscales indicated that the NICU children had a shorter spatial memory span length and committed more forgetting errors on a spatial working memory task. Correlational analyses demonstrated that the number and extent of medical complications at birth was negatively associated with spatial memory span, planning and spatial recognition memory. Multiple regression models suggested that gestational age was of primary importance in predicting spatial memory span, while neurobiological risk was primary in the prediction of spatial working memory errors. Overall, the current results showed fewer deficits in this group of children than were found in a previous neuropsychological assessment of this cohort. The implications of this finding for discerning the effects of neural plasticity over and above normal brain maturational processes are discussed. PMID- 12077014 TI - The antalgic efficacy of chronic motor cortex stimulation is related to sensory changes in the painful zone. AB - Epidural motor cortex stimulation (MCS) could achieve good pain control in patients with drug-resistant chronic neurogenic pain. In the search for parameters associated with the favourable outcome of this surgical procedure, quantitative sensory testing was performed in a series of 31 patients treated by MCS for chronic pain. Non-nociceptive and nociceptive sensory thresholds were measured in the painful area and its contralateral homologous zone with the stimulator in 'off' and in 'on' position. All 13 patients who exhibited normal or quite normal non-nociceptive thermal thresholds within the painful area benefited from MCS. Of the remaining 18 patients with altered thermal sensory thresholds, eight patients nevertheless experienced good pain control by MCS. In these eight 'good responders', sensory thresholds were improved by switching 'on' MCS. In contrast, the last 10 patients showed abnormal thermal thresholds that were not modified by switching 'on' MCS, and did not respond clinically to MCS. Therefore, 'good responders' to MCS could be identified by the absence of alteration of non nociceptive sensory modalities within the painful area, or by abnormal sensory thresholds that could be improved by MCS. These results additionally suggest that MCS acts on neural pathways involved in sensory discrimination that, in turn, are able to modulate the transmission of pain signals. PMID- 12077015 TI - Defining neurodegenerative diseases. PMID- 12077016 TI - Treating neurodegenerative diseases. PMID- 12077017 TI - Neurodegeneration in the age of molecular biology. PMID- 12077018 TI - The wall between neurology and psychiatry. PMID- 12077019 TI - Demystifying neurology. PMID- 12077020 TI - Iatrogenic stigma of mental illness. PMID- 12077021 TI - Diagnosing brain death without a neurologist. PMID- 12077022 TI - Patient safety agency admits problems with its pilot scheme. PMID- 12077023 TI - Government makes U turn on private practice ban. PMID- 12077029 TI - RCN general secretary may face "no confidence" vote. PMID- 12077027 TI - beta Blockers protect brain during cardiac surgery. PMID- 12077030 TI - An exceptional man. PMID- 12077031 TI - First hundred cases of variant Creutzfeldt-Jakob disease: retrospective case note review of early psychiatric and neurological features. AB - OBJECTIVE: To describe the early psychiatric and neurological features of variant Creutzfeldt-Jakob disease. DESIGN: Cohort study. SETTING: National surveillance system for Creutzfeldt-Jakob disease in the United Kingdom. PARTICIPANTS: The first 100 cases of variant Creutzfeldt-Jakob disease identified in the United Kingdom. MAIN OUTCOME MEASURES: The timing and nature of early psychiatric and neurological symptoms in variant Creutzfeldt-Jakob disease. RESULTS: The early stages of variant Creutzfeldt-Jakob disease are dominated by psychiatric symptoms, but neurological symptoms precede psychiatric symptoms in 15% of cases and are present in combination with psychiatric symptoms in 22% of cases from the onset of disease. Common early psychiatric features include dysphoria, withdrawal, anxiety, insomnia, and loss of interest. No common early neurological features exist, but a significant proportion of patients do exhibit neurological symptoms within 4 months of clinical onset, including poor memory, pain, sensory symptoms, unsteadiness of gait, and dysarthria. CONCLUSIONS: Although the diagnosis of variant Creutzfeldt-Jakob disease may be impossible in the early stages of the illness, particular combinations of psychiatric and neurological features may allow early diagnosis in an appreciable proportion of patients. PMID- 12077032 TI - Sleep attacks in patients taking dopamine agonists: review. AB - OBJECTIVES: To assess the evidence for the existence and prevalence of sleep attacks in patients taking dopamine agonists for Parkinson's disease, the type of drugs implicated, and strategies for prevention and treatment. DESIGN: Review of publications between July 1999 and May 2001 in which sleep attacks or narcoleptic like attacks were discussed in patients with Parkinson's disease. RESULTS: 124 patients with sleep events were found in 20 publications. Overall, 6.6% of patients taking dopamine agonists who attended movement disorder centres had sleep events. Men were over-represented. Sleep events occurred at both high and low doses of the drugs, with different durations of treatment (0-20 years), and with or without preceding signs of tiredness. Sleep attacks are a class effect, having been found in patients taking the following dopamine agonists: levodopa (monotherapy in 8 patients), ergot agonists (apomorphine in 2 patients, bromocriptine in 13, cabergoline in 1, lisuride or piribedil in 23, pergolide in 5,) and non-ergot agonists (pramipexole in 32, ropinirole in 38). Reports suggest two distinct types of events: those of sudden onset without warning and those of slow onset with prodrome drowsiness. CONCLUSION: Insufficient data are available to provide effective guidelines for prevention and treatment of sleep events in patients taking dopamine agonists for Parkinson's disease. Prospective population based studies are needed to provide this information. PMID- 12077033 TI - Randomised double blind trial of etoricoxib and indometacin in treatment of acute gouty arthritis. AB - OBJECTIVE: To assess the safety and efficacy of etoricoxib, a selective cyclo oxygenase-2 inhibitor, in comparison with indometacin in the treatment of acute gouty arthritis. DESIGN: Randomised, double blind, active comparator controlled trial. SETTING: 43 outpatient study centres in 11 countries. PARTICIPANTS: 142 men and eight women (75 patients per treatment group) aged 18 years or over presenting with clinically diagnosed acute gout within 48 hours of onset. INTERVENTIONS: Etoricoxib 120 mg administered orally once daily versus indometacin 50 mg administered orally three times daily, both for 8 days. MAIN OUTCOME MEASURES: Patients' assessment of pain in the study joint over days 2 to 5 (primary end point); investigators' and patients' global assessments of response to treatment and tenderness of the study joint (key secondary end points). RESULTS: Etoricoxib showed efficacy comparable to indometacin. Patients' assessment of pain in the study joint (0-4 point Likert scale, "no pain" to "extreme pain") over days 2 to 5 showed a least squares mean change from baseline of -1.72 (95% confidence interval -1.90 to -1.55) for etoricoxib and -1.83 (-2.01 to -1.65) for indometacin. The difference between treatment groups met prespecified comparability criteria. All other efficacy end points, including those reflecting reduction in inflammation and analgesia, provided corroborative evidence of comparable efficacy. Significant pain relief was evident at the first measurement, 4 hours after the first dose of treatment. Prespecified safety analyses revealed that drug related adverse experiences occurred significantly less frequently with etoricoxib (22.7%) than with indometacin (46.7%) (P=0.003), although overall adverse experience rates were similar between the two treatment groups. CONCLUSION: Etoricoxib 120 mg once daily provides rapid and effective treatment for acute gouty arthritis comparable to indometacin 50 mg three times daily. Etoricoxib was generally safe and well tolerated in this study. PMID- 12077034 TI - Comparison of physician outcome measures and patients' perception of benefits of inpatient neurorehabilitation. PMID- 12077035 TI - Locomotor milestones and babywalkers: cross sectional study. PMID- 12077036 TI - Prevalence of epilepsy in prisoners: systematic review. PMID- 12077037 TI - Mental health problems of children in the community: 18 month follow up. PMID- 12077038 TI - Drug points: Severe myalgia from an interaction between treatments with pantoprazole and methotrexate. PMID- 12077040 TI - 10-minute consultation: Paraesthesia. PMID- 12077039 TI - Reliability of N-terminal pro-brain natriuretic peptide assay in diagnosis of heart failure: cohort study in representative and high risk community populations. AB - OBJECTIVE: To investigate the performance of a novel assay for N-terminal pro brain natriuretic peptide (NT-proBNP) in diagnosing heart failure in various randomly selected general and high risk community populations. DESIGN: Community cohort study (substudy of the echocardiographic heart of England screening study). SETTING: Four randomly selected general practices in the West Midlands of England. PARTICIPANTS: 591 randomly sampled patients over the age of 45, stratified for age and socioeconomic status and falling into four cohorts (general population, patients with an existing clinical label of heart failure, patients prescribed diuretics, and patients deemed at high risk of heart failure). MAIN OUTCOME MEASURE: Sensitivity, specificity, positive and negative predictive values, likelihood ratios, and area under receiver operating characteristic curve for NT-proBNP assay in the diagnosis of heart failure. RESULTS: For NT-proBNP in the diagnosis of heart failure in the general population (population screen), a level of >36 pmol/l had a sensitivity of 100%, a specificity of 70%, a positive predictive value of 7%, a negative predictive value of 100%, and an area under the receiver operating characteristic curve of 0.92 (95% confidence interval 0.82 to 1.0). Similar negative predictive values were found for patients from the three other populations screened. CONCLUSIONS: This NT-proBNP assay seems to have value in the diagnosis of heart failure in the community. High negative predictive values indicate that the assay's chief use would be to rule out heart failure in patients with suspected heart failure with normal concentrations of NT-proBNP. Positive results may identify patients who need cardiac imaging. PMID- 12077041 TI - What we need to know about age related memory loss. PMID- 12077042 TI - Lesson of the week: Akathisia: overlooked at a cost. PMID- 12077043 TI - Evaluating drug treatments for Parkinson's disease: how good are the trials? PMID- 12077044 TI - Folic acid, ageing, depression, and dementia. PMID- 12077045 TI - Leprosy elimination-a virtual phenomenon or a reality? PMID- 12077046 TI - Discontinuation of thioridazine in patients with learning disabilities: balancing cardiovascular toxicity with adverse consequences of changing drugs. PMID- 12077047 TI - All you need is cognitive behaviour therapy? Critical appraisal of evidence base must be understood and respected. PMID- 12077048 TI - The limits of psychiatry. Psychospirituality will benefit heathcare staff. PMID- 12077049 TI - Renal transplantation. Spain's system for procuring organs operates in hospitals. PMID- 12077050 TI - Africa can solve its own health problems. Africa needs less globalisation and more real assistance . . . PMID- 12077051 TI - More units dedicated to women presenting with miscarriage are needed. PMID- 12077052 TI - Simple dosage guide for suncreans will help users. PMID- 12077053 TI - Proceedings of the 1st International Symposium on Visceral Perception in Gastroenterology. PMID- 12077054 TI - Neuroanatomy of visceral nociception: vagal and splanchnic afferent. AB - Afferent fibres convey sensory information from the upper gastrointestinal tract to the central nervous system but the nature of this information is different for vagal and spinal pathways. Vagal afferents convey predominantly physiological information while spinal afferents are able to encode noxious events. Because of the different response profiles following activation of these pathways, it is likely that vagal and splanchnic afferents play different roles in mediating sensation. PMID- 12077055 TI - Prevertebral ganglia and intestinofugal afferent neurones. AB - Intestinofugal afferent neurones (IFANs) are a unique subset of myenteric ganglion neurones that regulate normal gastrointestinal function. The IFANs relaying mechanosensory information to sympathetic neurones of the prevertebral ganglion (PVG) function as volume detectors. It is possible that mechanosensory information arriving in the PVG via axon collaterals of visceral spinal afferent nerves can be modulated entirely within the PVG itself. PMID- 12077056 TI - Visceral perception: sensory transduction in visceral afferents and nutrients. AB - The possible mechanisms that may be involved in nutrient detection in the wall of the gastrointestinal tract are reviewed. There is strong functional and electrophysiological evidence that both intrinsic and extrinsic primary afferent neurones mediate mechano- and chemosensitive responses in the gastrointestinal tract. This review focuses on the extrinsic afferent pathways as these are the ones that convey information to the central nervous system which is clearly necessary for perception to occur. PMID- 12077057 TI - Gastric hyperalgesia and changes in voltage gated sodium channel function in the rat. AB - Potential peripheral mechanisms that might contribute to the development of visceral hypersensitivity and ultimately to the altered sensations that characterise functional gastrointestinal disorders were examined by developing experimental models of gastric hyperalgesia. A direct link between alteration in behaviour and potential mechanisms of increased excitability of the gastric afferent innervation was found. PMID- 12077058 TI - Visceral perception: inflammatory and non-inflammatory mediators. AB - Visceral hypersensitivity is currently the most widely accepted mechanism responsible for abdominal pain. Inflammatory mediators are known to sensitise primary afferents and to recruit silent nociceptors. Recent evidence suggests that non-inflammatory mediators also have the potential to trigger visceral pain. This sequence of events may constitute part of an alerting system which prompts the central nervous system to correct gastrointestinal responses to ingestion. PMID- 12077060 TI - Hypersensitivity in functional gastrointestinal disorders. AB - Patients with functional gastrointestinal disorders may have visceral sensory dysfunction so that physiological stimuli induce their symptoms. The clinical significance of altered perception-that is, its relation to clinical symptoms remains unclear. Data indicate that sensory dysfunction is associated with altered reflex activity. Hence evidence of combined sensory-reflex dysfunction as a common pathophysiological mechanism in various functional gastrointestinal disorders would suggest that they are different forms of the same process. Altered reflex activity and altered conscious perception of gastrointestinal stimuli may combine to differing degrees, and their interaction may produce clinical symptoms. PMID- 12077061 TI - Role of the brain and sensory pathways in gastrointestinal sensory disorders in humans. AB - Several features of the irritable bowel syndrome (IBS) suggest involvement of the emotional limbic system in the brain. Abnormalities which upregulate afferent (sensory) signal intensity anywhere in this system, from the gastrointestinal tract to the brain, could induce hypersensitivity, leading to the pain and discomfort that characterise IBS and other functional disorders. Functional gastrointestinal disorders are likely to be heterogeneous given the complexity of the afferent system, and a number of different perturbations are possible. Intestinal hypersensitivity to pain and discomfort and associated reflex alterations in motility might explain the symptoms of functional bowel diseases. PMID- 12077062 TI - Testing the sensitivity hypothesis in practice: tools and methods, assumptions and pitfalls. AB - Visceral hypersensitivity is widely regarded as the reason for the development of functional gastrointestinal disorders, including functional dyspepsia and irritable bowel syndrome. The principles and techniques involved in testing the hypothesis that visceral sensitivity is important are discussed, together with the controversies in the assumptions, methods, and interpretations of the data acquired to date. PMID- 12077063 TI - A role for inflammation in irritable bowel syndrome? AB - Attention has been directed to the putative role of low grade mucosal inflammation in irritable bowel syndrome (IBS) on the basis of evidence showing that some patients with IBS have an increased number of inflammatory cells in the colonic and ileal mucosa. Previous episodes of infectious enteritis, genetic factors, undiagnosed food allergies, and changes in bacterial microflora may all play a role in promoting and perpetuating this low grade inflammatory process. Human and animal studies support the concept that inflammation may perturb gastrointestinal reflexes and activate the visceral sensory system even when the inflammatory response is minimal and confined to the mucosa. Thus abnormal neuroimmune interactions may contribute to the altered gastrointestinal physiology and hypersensitivity that underlies IBS. A brief review of the human and animal studies that have focused on the putative role of intestinal inflammation and infections in the pathogenesis of IBS is given. PMID- 12077065 TI - Food and hypersensitivity in functional dyspepsia. AB - It has long been known that stress affects both the stomach and colon, as shown by the very high prevalence of gastrointestinal symptoms among patients with psychiatric illness. The source may be limbic or peripheral, involving encoded memories or physiological changes. It is important to realise that physical symptoms such as those of functional dyspepsia do not only mean that the patient has a stomach disorder which needs to be identified and treated with specific pharmacological remedies, they often represent, in metaphorical and symbolic form, a state of disharmony brought about a specific psychosocial situation. It is only when that situation is understood and acknowledged that the patient can begin to get better. PMID- 12077064 TI - Role of thyrotrophin releasing hormone and corticotrophin releasing factor in stress related alterations of gastrointestinal motor function. AB - There is a growing body of experimental and clinical evidence to indicate that stress influences gastrointestinal motility. The most common pattern of gastrointestinal motor alterations induced by a variety of different stress factors is that of delayed gastric emptying and accelerated colonic transit. Central administration of corticotrophin releasing factor mimics both of these effects. This review focuses on the effects of two centrally acting peptides known to influence gastrointestinal motility and transit in experimental animals: thyrotrophin releasing hormone and corticotrophin releasing factor. The biological actions of these peptides are discussed in relation to the motility changes and pathways involved in their actions. PMID- 12077066 TI - The role of fat and cholecystokinin in functional dyspepsia. AB - The main factors involved in the pathophysiology of fat induced dyspepsia were investigated by reviewing a series of controlled double blind randomised studies which sought to determine the role of nutrient fat and the postprandial release of cholecystokinin (CCK) in the development of dyspeptic symptoms in healthy volunteers and in patients with functional dyspepsia. The studies showed that during distension of the stomach, lipids are a major trigger of dyspeptic symptoms such as nausea, bloating, pain, and fullness, and that they modulate upper gastrointestinal sensations and symptoms in a dose related fashion. CCK is a major mediator of the sensitisation of gastric perception by lipids in patients with functional dyspepsia as the CCK-A receptor antagonist dexloxiglumide markedly diminishes this effect. The studies provide important insights into the mechanisms underlying gastrointestinal perception in response to fat and the role of CCK in patients with functional dyspepsia. PMID- 12077068 TI - Non-invasive measurement of gastric accommodation in humans. AB - Gastric accommodation describes the reduction in gastric tone and increase in compliance that follows ingestion of a meal and involves at least two responses: "receptive relaxation" which allows the stomach to accept a volume load without a significant rise in gastric pressure and "adaptive relaxation" which modulates gastric tone in response to the specific properties of the meal ingested. However, there are considerable technical difficulties in measuring the accommodation process. The current standard barostat studies, and other methods such as conventional and three dimensional ultrasound, or single photon emission computed tomography have significant disadvantages. Preliminary findings from the development and validation of a new magnetic resonance imaging technique that addresses many of the deficiencies of previous methods are presented. PMID- 12077069 TI - Pathophysiology of functional dyspepsia. AB - Functional dyspepsia is a symptom complex characterised by postprandial upper abdominal discomfort or pain, early satiety, nausea, vomiting, abdominal distension, bloating, and anorexia in the absence of organic disease. Gastrointestinal motor abnormalities, altered visceral sensation, and psychosocial factors have all been identified as major pathophysiological mechanisms. This perspective has now replaced the earlier view that the condition was the result of a sole motor or sensory disorder of the stomach. Future therapeutic strategies should be aimed at reducing nociception as well as enhancing the accommodation response. PMID- 12077070 TI - Role of visceral sensitivity in the pathophysiology of irritable bowel syndrome. AB - Visceral hypersensitivity has been recognised as a characteristic of patients with irritable bowel syndrome (IBS). It may be involved in the pathogenesis of abdominal pain/discomfort, and seems to result from the sensitisation of nerve afferent pathways originating from the gastrointestinal tract. From a clinical point of view, hypersensitivity, although frequent, is not a constant finding among patients with IBS and cannot therefore be considered as a diagnostic marker of the condition. The advances made in understanding visceral hypersensitivity in patients with IBS are reviewed: the factors that influence abdominal distension are defined and different therapeutic perspectives are examined. PMID- 12077071 TI - Sensation and gas dynamics in functional gastrointestinal disorders. AB - Our current knowledge of motor and sensory functions in the human gut is critically reviewed, showing how the two may interact to produce symptoms in patients with functional gastrointestinal disorders. A local stimulus is necessary to activate the pathogenetic symptom generation process, and in many patients abnormal pooling of gas at various or extensive sites in the bowel and focal gut distension may provide the local stimulus, compounded by spatial summation phenomena and conscious visceral hypersensitivity. The interplay of these mechanisms results in the clinical expression of symptoms. PMID- 12077073 TI - Serotonergic modulation of visceral sensation: upper gastrointestinal tract. AB - Agents that modify serotonergic function have therapeutic potential for the treatment of visceral hypersensitivity, either through a direct effect on perception or through modulation of visceral tone or motility. Administration of selective serotonin reuptake inhibitors reduces oesophageal sensitivity to distension but not gastric sensitivity to distension. 5-HT ligands may also influence gastric mechanosensitivity by altering tone. Although the exact role of 5-HT receptors in the control of gastrointestinal functions remains unknown, 5-HT is generally considered to be the main candidate involved in the modulation of motor and sensory function from the gastrointestinal tract. Hence serotonergic modulation of upper gut sensitivity appears to be promising for the development of novel approaches to the treatment of functional disorders of the upper gastrointestinal tract. PMID- 12077074 TI - Serotonergic modulation of visceral sensation: lower gut. AB - The role of 5-HT agents in the modulation of lower gastrointestinal function is discussed. Selective serotonin reuptake inhibitors are of potential benefit in functional gastrointestinal diseases although formal evidence is lacking. Novel pharmacological approaches include 5-HT(3) antagonists and 5-HT(4) agonists. These pharmacological classes have shown beneficial effects on a global efficacy end point, and ameliorated more than one symptom of lower gut function in clinical trials. They offer promise for the development of novel therapies for the treatment and control of irritable bowel syndrome. PMID- 12077075 TI - Pharmacotherapy: non-serotonergic mechanisms. AB - Antidepressants rapidly relieve pain in irritable bowel syndrome (IBS) and are effective at low doses. Noradrenaline reuptake inhibitors appear to be more effective than selective serotonergic reuptake inhibitors, suggesting that pathways other than those modulated by serotonin may be involved in visceral sensation. Visceral sensitivity is reduced by both centrally and peripherally acting opioids, suggesting the possible existence of an endogenous opioid deficiency in patients with IBS. The alpha(2) adrenoceptor antagonist clonidine, as well as somatostatin, oxytocin, and possibly amitriptyline have also been shown to act as visceral analgesics. As knowledge increases, there are undoubtedly many other possible targets, and new drugs currently undergoing development may provide future benefit in patients with IBS. PMID- 12077078 TI - Autoimmune related pancreatitis. AB - Since the first documented case of a particular form of pancreatitis with hypergammaglobulinaemia, similar cases have been reported, leading to the concept of an autoimmune related pancreatitis or so-called "autoimmune pancreatitis". Although it has not yet been widely accepted as a new clinical entity, the present article discusses the recent concept of autoimmune pancreatitis. PMID- 12077076 TI - Centrally acting agents and visceral sensitivity. AB - The evidence relating to the site and mechanism of action of "centrally acting" agents which may affect visceral sensitivity is reviewed. Antidepressant drugs such as amitriptyline as well as the newer selective serotonin reuptake inhibitors are thought to act at the level of the CNS. Opiates, including morphine as well as compounds such as trimebutine or fedotozine designed for therapeutic use in irritable bowel syndrome, are effective in reducing visceral nociception. Cytokines in the CNS are known to be involved in the modulation of pain and there is also evidence to suggest that centrally acting cytokines may play a role in the production of visceral hypersensitivity. Consequently, they may provide an interesting target for future research. PMID- 12077079 TI - A place for TACE. PMID- 12077080 TI - A long hard look at Dukes' B. PMID- 12077081 TI - Weighty issues in hepatitis C. PMID- 12077082 TI - To perform or not to perform liver biopsy-that is the question. PMID- 12077083 TI - Ileal Crohn's disease is best treated by surgery. PMID- 12077084 TI - Ileal Crohn's disease is best treated by surgery. PMID- 12077085 TI - Risk factors for ulcer development in patients with non-ulcer dyspepsia: a prospective two year follow up study of 209 patients. AB - BACKGROUND AND AIMS: A subset of non-ulcer dyspepsia (NUD) disorders can evolve into peptic ulcer disease. This prospective study attempted to determine the independent risk factors for ulcer formation in NUD patients, and compared the natural history of Helicobacter pylori positive and negative NUD subjects. METHODS: From May 1997 to April 1999, consecutive NUD patients were enrolled into the study. Endoscopy was performed routinely on enrolment, at the end of the second and 12th months, and whenever there was a dyspepsia attack. Patients were prospectively followed up for two years. RESULTS: Peptic ulcers occurred in 16 of 209 NUD patients during the two year follow up period. Multivariate analysis of 13 host and bacterial factors demonstrated that advanced age (odds ratio 2.90), H pylori infection (odds ratio 3.59), and use of non-steroidal anti-inflammatory drugs (NSAID; odds ratio 4.46) were independently significant in predicting subsequent peptic ulcer development. NUD patients with all three risk factors had a 75% (3/4) risk of developing peptic ulcer but the ulcer incidence in patients without any of the risk parameters was only 1.2% (1/84). The resolution rate of symptoms in the H pylori positive NUD patients was similar to the H pylori negative patients (57.9% v 49.1%; 95% confidence interval (CI) -5 to 22). However, rates for subsequent peptic ulcer and erosion development were significantly higher in H pylori positive patients than in H pylori negative patients (ulcer 12.6% v 3.5%, 95% CI 1-16; erosion 23.2% v 12.3%, 95% CI 1-21). CONCLUSION: A small but significant proportion of NUD patients develop peptic ulcer after long term follow up. H pylori infection, NSAID use, and advanced age are independent risk factors for subsequent ulcer formation. Follow up endoscopy is strongly indicated for an NUD patient with multiple risk factors for ulcer formation when symptoms recur. PMID- 12077087 TI - Adult height in patients with early onset of Crohn's disease. AB - BACKGROUND: Growth impairment during childhood and adolescence is a common problem faced by patients with an early onset of Crohn's disease. AIMS: To establish how the final adult height is affected in patients with early onset of symptoms of Crohn's disease. METHODS: Information on height, parental height, and disease history was obtained from 135 patients with Crohn's disease who reached their adult height (men 22-40 years, women 18-40 years) using a questionnaire and by outpatient measurement of height where possible. Subsequently, adult heights were expressed as standard deviation scores, with and without correction for the expected target height. RESULTS: Patients with onset of disease before puberty were shorter compared with patients with onset in adulthood (p<0.01). This difference was not statistically significant when adult heights were corrected for parental height. Also, height standard deviation scores for those patients with onset of disease before puberty were significantly lower than those with onset of disease during puberty (p<0.05) but after correction for parental height the difference was not significant. The site of disease had no influence on adult height. Patients who had used corticosteroids during puberty were significantly shorter than patients who had not (p=0.005). This was also true when corrected for target height (p=0.007). CONCLUSIONS: Although there was a trend indicating a deficit in adult height in patients with an early onset of Crohn's disease, once adjustment was made for parental height, this difference was not significant. Use of corticosteroids in puberty resulted in lower adult height. PMID- 12077086 TI - In siblings with similar genetic susceptibility for inflammatory bowel disease, smokers tend to develop Crohn's disease and non-smokers develop ulcerative colitis. AB - BACKGROUND AND AIMS: Smoking tobacco has opposite effects on the different forms of inflammatory bowel disease (IBD). It predisposes to the development of Crohn's disease (CD) yet is associated with a reduced incidence of ulcerative colitis (UC). We have studied sib pairs discordant for both smoking and IBD phenotype (UC or CD) to investigate whether smoking determines the type of IBD that develops in individuals with very similar genetic susceptibility. PATIENTS: Smoking habits and disease characteristics were analysed in 242 IBD pedigrees (658 patients). Within this group there were 339 affected sibling pairs of whom 89 were discordant for smoking when diagnosed. RESULTS: Smoking at diagnosis was associated with development of CD (odds ratio (OR) 3.55; 95% confidence limits 2.50-5.02; p<0.001) in all of the familial patients, with increases when analysed for ileocaecal disease, fibrostenosis, and intestinal resection. Smokers were also protected from UC (OR 0.28; 0.2-0.4; p<0.001). Of 89 sibling pairs discordant for smoking at diagnosis, 23 were also discordant for disease type-in 21 of these, CD occurred in the smoker and UC in the non-smoker (OR 10.5; 2.6-92; p<0.0001). CONCLUSIONS: Smoking is a strong environmental risk factor for Crohn's disease and increases the likelihood of needing surgery. However, sib pairs who are discordant for both smoking and IBD type almost always show CD in the smoker and UC in the non-smoker, and so in some cases tobacco consumption acts on IBD genetic predisposition to shift the phenotype from UC towards CD. The explanation of part of the apparent "protective" effect of smoking on sporadic UC may be that the form of IBD that develops in a proportion of smokers is not UC but CD. PMID- 12077088 TI - Double blind, placebo controlled trial of the remission inducing and steroid sparing properties of an ICAM-1 antisense oligodeoxynucleotide, alicaforsen (ISIS 2302), in active steroid dependent Crohn's disease. AB - BACKGROUND AND AIMS: To evaluate the safety and efficacy of the intercellular adhesion molecule 1 (ICAM-1) antisense phosphorothioate oligonucleotide alicaforsen (ISIS 2302) in Crohn's disease. METHODS: Active (Crohn's disease activity index (CDAI) 200-350), steroid dependent (prednisone 10-40 mg) Crohn's patients were randomised into three treatment groups: placebo versus ISIS 2302 (2 mg/kg intravenously three times a week) for two or four weeks. Patients were treated in months 1 and 3, with steroid withdrawal attempted by week 10. The primary end point (steroid free remission) was a CDAI <150 off steroids at the end of week 14. RESULTS: A total of 299 patients were enrolled, with a mean baseline CDAI of 276 and steroid dose of 23 mg/day. Rates of steroid free remission were equivalent for the two and four week ISIS 2302 groups (20.2% and 21.2%) and the placebo group (18.8%). At week 14, steroid withdrawal was successful in more ISIS 2302 patients compared with placebo treated patients (78% v 64%; p=0.032). Steroid free remission was highly correlated with exposure (p=0.0064). Other clinical responses were correlated with exposure, with significant results versus placebo being observed in the highest area under the curve subgroup. CDAI scores decreased by 136 (112) at week 14 versus 52 (107) for placebo (p=0.027) and inflammatory bowel disease score questionnaire improved by 43 (31) versus 15 (36) for placebo (p=0.027). CONCLUSIONS: Although the primary outcomes failed to demonstrate efficacy, pharmacodynamic modelling suggests that alicaforsen (ISIS 2302) may be an effective therapy for steroid dependent Crohn's disease. PMID- 12077089 TI - Tumour necrosis factor alpha converting enzyme (TACE) activity in the colonic mucosa of patients with inflammatory bowel disease. AB - BACKGROUND: Anti-tumour necrosis factor alpha (TNF-alpha) antibodies are effective in Crohn's disease and perhaps ulcerative colitis but antigenicity and the high cost have raised interest in other strategies to block TNF-alpha. These include the TNF-alpha converting enzyme (TACE) which releases soluble TNF-alpha from transmembrane pro-TNF-alpha. AIM: To investigate whether TACE activity is present in human colonic mucosa. MATERIALS AND METHODS: Detergent extracts of cell membranes from colonic biopsies were obtained from 12 controls and 28 patients with inflammatory bowel disease. Enzyme activity was measured by hydrolysis assays using pro-TNF-alpha or oligopeptide substrates spanning the known pro-TNF-alpha cleavage site at Ala(76)-Val(77). Cleavage products were identified by western blotting, high pressure liquid chromatography, or mass spectrometry. TACE protein was localised by immunohistochemistry and identified by western blotting of detergent extracts from purified lamina propria mononuclear cells (LPMNC) or epithelial cells. RESULTS: Detergent extracts released TNF-alpha from pro-TNF-alpha and cleaved a model oligopeptide as predicted. Substrate hydrolysis was sensitive to known TACE/matrix metalloproteinase (MMP) inhibitors, but not trocade which has low activity against TACE. The median TACE level was increased in active ulcerative colitis (147 arbitrary units (AU)/mg; p<0.01) but not in Crohn's disease (81 AU/mg) compared with controls (79 AU/mg). Both the full length proform and the active form of TACE protein were expressed in LPMNC cells and epithelial cells. CONCLUSIONS: Functional TACE activity is ubiquitously expressed in the human colon and increased in ulcerative colitis, raising interest in MMP inhibitors targeting TACE. PMID- 12077090 TI - Related IgA1 and IgG producing cells in blood and diseased mucosa in ulcerative colitis. AB - BACKGROUND: Ulcerative colitis (UC) is a chronic inflammatory bowel disease in which the colonic mucosa is infiltrated with plasma cells producing IgG autoantibodies. It is not known whether this represents a local mucosal response which has switched to IgG or a peripheral response which may have been initiated by peripheral antigen which homed to the colonic mucosa. The clonal distribution of IgG secreting cells and isotype switched variants in UC is not known. AIMS: To investigate the clonal distribution of mucosal IgG in UC and to search for related IgG and IgA secreting cells in normal and diseased mucosa and blood in UC. To investigate characteristics which may discriminate between the mucosal and peripheral repertoire in the normal mucosa and in UC. PATIENTS: Blood and normal and diseased mucosa from two patients with UC were studied. METHODS: Immunoglobulin gene analysis and clone specific polymerase chain reaction were used to study the clonal distribution and characteristics of IgG and related IgA in the mucosa and blood of patients with UC. RESULTS: The IgG response in the mucosa of UC patients included widespread clones of cells that were present in both the diseased mucosa and blood but that were scarce in normal mucosa. Clonally related IgA class switch variants, all IgA1, were detected but also only in the diseased mucosa and blood. This suggests that these clones home preferentially to the diseased mucosa. We showed that J(H)1 usage was characteristic of the peripheral repertoire, and that examples of J(H)1 usage were observed in mucosal IgG in UC. CONCLUSIONS: Overall, these data are consistent with a model of UC in which a peripheral response is expressed and expanded in the colonic mucosa. PMID- 12077092 TI - Frequent loss of SMAD4/DPC4 protein in colorectal cancers. AB - BACKGROUND AND AIMS: Loss of DNA sequences from chromosome 18q21 is a major genetic change in colorectal tumorigenesis. Multiple genes have been identified in this area. One of these, DPC4 (deleted in pancreatic cancer 4, also known as SMAD4), is mutated in a minor subset of colorectal carcinomas as well as in germlines of humans predisposed to colon tumours. PATIENTS AND METHODS: The involvement of SMAD4 in sporadic colorectal neoplasia was evaluated by immunohistochemistry in 53 unselected cases and 27 cases displaying microsatellite instability. RESULTS: SMAD4 expression was absent in 20 of 53 (38%) unselected colorectal carcinomas, and reduced in another 15 (28%) cases. However, 26 of 27 cancers displaying microsatellite instability and TGF-betaIIR mutations were positive for SMAD4 immunostaining. CONCLUSIONS: Loss of SMAD4 expression may play a more prominent role in colon cancer than anticipated based on genetic evidence, but not in mutator phenotype tumours. PMID- 12077093 TI - Colon cancer in France: evidence for improvement in management and survival. AB - BACKGROUND: Cancer registries recording all cases diagnosed in a well defined population represent the only way to assess real changes in the management of colon cancer at the population level. AIMS: To determine trends over a 23 year period in treatment, stage at diagnosis, and prognosis of colon cancer in the Cote-d'Or region, France. PATIENTS: A total of 3389 patients with colon cancer diagnosed between 1976 and 1998. METHODS: Time trends in clinical presentation, surgical treatment, chemotherapy treatment, stage at diagnosis, postoperative mortality, and survival were studied. A non-conditional logistic regression was performed to obtain an odds ratio for each period adjusted for the other variables. To estimate the independent effect of the period on prognosis, a relative survival analysis was performed. RESULTS: Between 1976 and 1991, the resection rate increased from 69.3% to 91.9% and then remained stable. This increase was particularly marked in the older age group (56.4% to 90.5%). The proportion of stage III patients treated with adjuvant chemotherapy rose from 4.1% for the 1989-1990 period to 45.7% for the 1997-1998 period. Over the 23 years of the study the proportion of stage I and II patients increased from 39.6% to 56.6%, associated with a corresponding decrease in the proportion of patients with advanced stages. Postoperative mortality decreased from 19.5% to 7.3%. This led to an improvement in five year relative survival (from 33.0% for the 1976 1979 period to 55.3% for the 1992-1995 period). CONCLUSIONS: Advances in the management of colon cancer have resulted in improving the prognosis of this disease. However, progress is still possible, particularly in the older age group. PMID- 12077091 TI - Aberrant composition of gut microbiota of allergic infants: a target of bifidobacterial therapy at weaning? AB - BACKGROUND: Recent data have outlined a relationship between the composition of the intestinal microflora and allergic inflammation, and demonstrated the competence of probiotics in downregulation of such inflammation. AIMS: Our aims were to characterise the relationship between gut microbes and the extent of allergic sensitisation and to assess whether the efficacy of bifidobacterial supplementation in the treatment of allergy could relate to modulation of the intestinal microbiota. METHODS: This randomised study included 21 infants with early onset atopic eczema of whom eight were intolerant (highly sensitised group (HSG)) and 13 tolerant (sensitised group (SG)) to extensively hydrolysed whey formula (EHF). In the SG, six were weaned to EHF without (placebo group (PG)) and seven to EHF with Bifidobacterium lactis Bb-12 supplementation (bifidobacteria treated group (BbG)). The faecal microflora of infants in the HSG was analysed only before weaning whereas in the SG the faecal microflora was analysed both before and after weaning. RESULTS: Infants in the HSG had greater numbers of lactobacilli/enterococci than those in the SG. Serum total IgE concentration correlated directly with Escherichia coli counts in all infants and with bacteroides counts in the HSG, indicating that the presence of these bacteria is associated with the extent of atopic sensitisation. The effect of supplementation was characterised as a decrease in the numbers of Escherichia coli and protection against an increase in bacteroides numbers during weaning. CONCLUSIONS: These data indicate that bifidobacterial supplementation appears to modify the gut microbiota in a manner that may alleviate allergic inflammation. Further studies are needed to confirm this conclusion. PMID- 12077094 TI - Identification of objective pathological prognostic determinants and models of prognosis in Dukes' B colon cancer. AB - BACKGROUND AND AIMS: There is a need for objective easily determined pathological prognostic parameters in Dukes' B colon carcinoma to allow selection of such patients for further treatment as the role of adjuvant chemotherapy for these patients remains unclear. This study was initiated to assess the influence of pathological factors on prognosis in an unselected prospective series of Dukes' B colonic cancer. METHODS: The Gloucester Colorectal Cancer study, established in 1988, recruited more than 1000 cases. Meticulous pathological assessment of the 268 Dukes' B colonic cancer resections in this series included evaluation of all pathological factors that could influence staging and prognosis. All patients entered a comprehensive follow up system. RESULTS: Four pathologically determined factors--peritoneal involvement, venous spread (both submucosal and extramural), spread to involve a surgical margin, and perforation through the tumour-were independent prognostic factors in multivariate analysis. Combining these four factors into a simple cumulative scoring system generated clinically useful prognostic groups. CONCLUSIONS: The cumulative prognostic index allows apportionment of patients with Dukes' B colon cancer into defined prognostic groups, which in turn could allow more objective selection of patients for adjuvant therapy, especially as part of clinical trials. PMID- 12077095 TI - Cholinergic and nitrergic interneurones in the myenteric plexus of the human colon. AB - BACKGROUND: Myenteric interneurones are involved in the reflexes that control the motility of the human colon. AIMS: The distribution of choline acetyltransferase (ChAT) and nitric oxide synthase (NOS) immunoreactivity in myenteric interneurones was investigated in this study. METHODS: DiI (1,1'- didodecyl 3,3,3',3'-indocarbocyanine perchlorate) was applied to the myenteric plexus of the human colon followed by organotypic culture. Retrogradely labelled neurones, with projections longer than motor neurones (>10 mm), were studied to exclude motor neurone populations. ChAT and NOS immunoreactivity was then determined in the interneurones. RESULTS: We found that 90% of interneurones projecting orally contained ChAT and none contained NOS. Ninety five per cent of descending interneurones were labelled with ChAT and/or NOS antisera; 46% contained NOS immunoreactivity alone, 20% contained ChAT immunoreactivity alone, and 29% contained both ChAT and NOS. Anally directed interneurones had significantly longer projections than orally projecting interneurones. CONCLUSIONS: Nearly all interneurones contain either NOS or ChAT immunoreactivity. Orally projecting interneurones are of two types: 90% contain ChAT alone and the remainder contain immunoreactivity for neither ChAT nor NOS. There are three main types of anally projecting interneurones: the largest, which contains NOS but not ChAT, and the two smaller classes which contain ChAT and NOS, and CHAT alone. PMID- 12077096 TI - Absorption and metabolism of octanoate by the rat colon in vivo: concentration dependency and influence of alternative fuels. AB - BACKGROUND: Compared with short and long chain fatty acids, medium chain fatty acids (MCFAs) have been shown to provide the highest colonic absorption of substrate carbon. Moreover, colonic epithelial cells fulfil their basic energy requirements as easily from MCFAs as from short chain fatty acids. AIMS: To further characterise octanoate as a colonic luminal substrate, we determined in vivo the influence of (i) substrate concentration and (ii) alternative luminal fuels, on rat colonic transport and metabolism. METHODS: Segments of rat proximal colon (8 cm) were cannulated and perfused for 100 min with (14)C labelled octanoate. The right colic vein was also cannulated and venous blood analysed for total (14)C, (14)CO(2), and metabolites by scintillation counting and high performance liquid chromatography. RESULTS: Tracer appearance in mesenteric blood stabilised after 20-40 minutes of perfusion. Increasing luminal octanoate concentrations from 2 to 40 mM resulted in linear increases in total carbon absorption. Maximum CO(2) production was reached near 10 mM. A substantial proportion of octanoate was absorbed without being metabolised (59-94%). The luminal presence of a mixture of alternative fuels had no influence on either octanoate transport or metabolism. CONCLUSIONS: This study demonstrated substantial concentration dependent colonic absorption of octanoate, rendering this MCFA a potential and much needed high energy substrate for patients with compromised small bowel function. Moreover, octanoate meets the basic energy requirements of colonic epithelial cells in vivo as well as butyrate. This study also demonstrates the divergence of in vitro and in vivo data regarding fatty acid absorption and metabolism in the colonic epithelium. PMID- 12077097 TI - Association between severity of type A hepatitis and nucleotide variations in the 5' non-translated region of hepatitis A virus RNA: strains from fulminant hepatitis have fewer nucleotide substitutions. AB - BACKGROUND: Type A hepatitis is still a considerable problem in both underdeveloped and developed countries. Why some patients progress to fulminant type A hepatitis and others do not is unclear. AIMS: To determine if nucleotide differences in the genome of hepatitis A virus (HAV) are responsible for the range of clinical severities, we analysed the 5' non-translated region (5'NTR) of the HAV genome, which has an internal ribosomal entry site and is important for cap independent translation of the viral message. METHODS: Serum samples from 84 Japanese patients with sporadic type A hepatitis from five distant regions of Japan, comprising 12 patients with fulminant hepatitis (FH), 13 with severe acute hepatitis (AHs), and 59 with acute hepatitis (AH), were examined for HAV RNA. The fragment between nucleotides 75 and 638 of the 5'NTR was amplified by reverse transcription-polymerase chain reaction, and the nucleotide sequence was determined by direct sequencing. RESULTS: Comparison of sequences of the 5'NTR revealed relatively fewer nucleotide substitutions in FH and AHs patients compared with the considerable sequence variations found in strains of AH. This tendency was most prominent between nucleotides 200 and 500. Strains from FH and AHs cases had fewer nucleotide substitutions (p<0.001) in this region. CONCLUSIONS: Nucleotide variations in the central portion of the 5'NTR of HAV may influence the severity of type A hepatitis. PMID- 12077098 TI - Effect of weight reduction on liver histology and biochemistry in patients with chronic hepatitis C. AB - BACKGROUND: Steatosis occurs in more than 50% of patients with chronic hepatitis C and is associated with increased hepatic fibrosis. In many of these patients the pathogenesis of steatosis appears to be the same as for patients with non alcoholic fatty liver disease-that is, related to visceral adiposity and obesity. METHODS: The effect of a three month weight reduction programme on liver biochemistry and metabolic parameters was examined in 19 subjects with steatosis and chronic hepatitis C. Paired liver biopsies were performed in 10 subjects, prior to and 3-6 months following the intervention, to determine the effect of weight loss on liver histology. RESULTS: There was a mean weight loss of 5.9 (3.2) kg and a mean reduction in waist circumference of 9.0 (5.0) cm. In 16 of the 19 patients, serum alanine aminotransferase levels fell progressively with weight loss. Mean fasting insulin fell from 16 (7) to 11 (4) mmol/l (p<0.002). Nine of 10 patients with paired liver biopsies had a reduction in steatosis irrespective of viral genotype. In these subjects the median modified Knodell fibrosis score decreased from 3 to 1 (p=0.04) and activated stellate cells significantly decreased (p<0.004). CONCLUSIONS: Weight loss in patients with chronic hepatitis C may be associated with a reduction in steatosis and abnormal liver enzymes and an improvement in fibrosis, despite the persistence of the virus. Weight reduction may provide an important adjunct treatment strategy for patients with chronic hepatitis C. PMID- 12077099 TI - Acute hepatitis C virus (HCV) infection in chronic carriers of hepatitis B virus (HBV): the impact of underlying active HBV replication on persistence of HCV infection and antibody responses to HCV. AB - BACKGROUND AND AIMS: The aim of this study was to assess whether underlying chronic hepatitis B virus (HBV) infection interferes with persistence of hepatitis C virus (HCV) infection and humoral immune responses to HCV in acute HCV infection. METHODS: Serial sera from 12 patients with acute HCV infection (group A) and 12 hepatitis B surface antigen (HBsAg) carriers with acute HCV infection (seven anti-hepatitis B e antigen (anti-HBe) positive (group B1) and five hepatitis B e antigen (HBeAg) positive (group B2)) were tested for HCV RNA by polymerase chain reaction, and anti-HCV by third generation enzyme immunoassay and confirmatory assay. Serial serum samples from HBsAg carriers were also tested for HBeAg, anti-HBe, and HBV DNA by hybridisation assay. RESULTS: Persistent HCV viraemia for more than six months was significantly more frequent in groups A (83%) and B1 (86%) than in group B2 (0%). Anti-HCV was detected in 100% and 86% of group A and group B1 one month after onset while only one group B2 patient was transiently anti-HCV positive 1-2 months after onset. Of the latter, three had anti-core 1 less than two months after onset while no patient responded to other HCV antigens. Overall, of six HBsAg carriers with acute self limiting HCV infection, only one had transient anti-HCV and three had transient anti-core 1. HBV DNA became undetectable transiently in four and persistently in one group B2 patient. CONCLUSION: The presence of active HBV replication can inhibit the persistence of HCV infection and antibody responses to HCV. Acute HCV infection in HBsAg carriers with active HBV replication usually presents transient HCV viraemia with poor antibody responses to HCV. PMID- 12077100 TI - Augmentation effect of postprandial hyperinsulinaemia on growth of human hepatocellular carcinoma. AB - BACKGROUND: Cirrhotic patients with hepatocellular carcinoma (HCC) frequently have impaired glucose metabolism. AIMS: To investigate whether impaired glucose metabolism affects the growth rate of the tumour. PATIENTS AND METHODS: Tumour doubling time (DT), assessed by ultrasound imaging analysis, was measured in 60 patients with single small HCC (diameter <30 mm). DT was compared with plasma insulin and glucose concentrations following the oral glucose tolerance test (OGTT). The effect of continuous infusion of octreotide (a somatostatin analogue 200 microg/day) for three months on DT in five cases was assessed. RESULTS: The 60 patients were divided into two groups because the median DT was 140 days: rapid growth group (DT 140 days, n=30). Fasting plasma insulin concentration and area under the plasma insulin curve (AUC(ins)) of the OGTT (10.4 (6.2) microU/ml and 262 (152) microU/ml/h, respectively; mean (SD)) in the rapid growth group were significantly higher than those in the slow growth group (7.6 (4.3) and 146 (140), respectively) (p=0.041 and p=0.0006, respectively). In contrast, fasting plasma glucose concentration and area under the plasma glucose curve (AUC(gluc)) in the rapid growth group were significantly lower than those in the slow growth group (p=0.0003 and p=0.0012, respectively). Univariate and multivariate analyses of logistic regression models demonstrated that AUC(ins) was a significant factor contributing to the growth rate of HCC (p=0.001 and p=0.016, respectively). AUC(ins) significantly decreased after octreotide treatment (p<0.02) but AUC(gluc) did not significantly change. DT after treatment increased in three of the five patients and could not be calculated in the remaining two patients because of no change in the diameter of the tumour. CONCLUSIONS: These data suggest that postprandial hyperinsulinaemia is associated with accelerated HCC growth. PMID- 12077101 TI - Prospective comparative study of spiral computer tomography and magnetic resonance imaging for detection of hepatocellular carcinoma. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is often detected at a relatively late stage when tumour size prohibits curative surgery. Screening to detect HCC at an early stage is performed for patients at risk. AIM: The aim of this study was to compare prospectively the diagnostic accuracy and classification for management of the two state of the art secondline imaging techniques: triphasic spiral computer tomography (CT) and super paramagnetic iron oxide (SPIO) enhanced magnetic resonance imaging (MRI). PATIENTS: Sixty one patients were evaluated between January 1996 and January 1998. Patients underwent CT and MRI within a mean interval of 6.75 days. METHODS: CT and MRI were evaluated blindly for the presence and number of lesions, characterisation of these lesions, and classification for management. For comparison of the data on characterisation, the CT and MRI findings were compared with histopathological studies of the surgical specimens and/or follow up imaging. Data of patients not lost to follow up were available to January 2001. RESULTS: SPIO enhanced MRI detected more lesions and overall smaller lesions than triphasic spiral CT (number of lesions 189 v 124; median diameter 1.0 v 1.8 cm; Spearman rank's correlation coefficient 0.63, p<0.001). There was no significant difference in accuracy between CT and MRI for lesion characterisation. The agreement in classification for management was very good (weighted kappa 0.91, 95% CI 0.83-0.99). CONCLUSION: SPIO enhanced MRI detects more and smaller lesions, but both techniques are comparable in terms of classification for management. SPIO enhanced MRI may be preferred as there is no exposure to ionising radiation. PMID- 12077102 TI - Underdiagnosis of hereditary haemochromatosis: lack of presentation or penetration? AB - BACKGROUND: The majority of hereditary haemochromatosis (HH) patients are homozygous for the C282Y mutation in the HFE gene. We have demonstrated a homozygote frequency of 1 in 83 for the C282Y mutation in a retrospective analysis of Irish neonates. However, a fully developed phenotype is not observed at the same frequency clinically, suggesting that a large proportion of Irish HH patients may remain undiagnosed. AIMS: To determine whether underdiagnosis of HH results from the non-specific nature of early symptoms or incomplete penetrance of the C282Y mutation. METHODS: Seventy nine C282Y homozygous individuals identified from family screening for HH and 30 HH probands were investigated. Non specific symptoms (fatigue, arthropathy, and impotence) and their association with iron indices (transferrin saturation and serum ferritin) and hepatic iron deposition were analysed. RESULTS: We found that 78% of men (mean age 42 years) and 36% of women (mean age 39 years) who were identified as C282Y homozygotes following family screening had iron overload, as defined by a transferrin saturation >or=52% combined with a serum ferritin >or=300 microg/l for men and >or=200 microg/l for women. The frequency of reports of non-specific symptoms in those individuals with iron overload was not significantly different from those who did not have iron overload. CONCLUSIONS: Our findings indicate that underdiagnosis of HH may be due to the non-specific nature of early symptoms and less frequently to the incomplete penetrance of the C282Y mutation. PMID- 12077104 TI - High frequency of autoantibodies in patients with primary sclerosing cholangitis that bind biliary epithelial cells and induce expression of CD44 and production of interleukin 6. AB - AIM: Sera of patients with autoimmune liver diseases were investigated for the presence of autoantibodies binding to human biliary epithelial cells (BECs). Furthermore, their functional capacity was investigated by testing their capacity to fix complement as well as induce expression of various adhesion molecules and production of cytokines. METHODS: Sera from patients with various stages of primary sclerosing cholangitis (PSC; n=30), primary biliary cirrhosis (PBC; n=29), autoimmune hepatitis (AIH; n=25), and normal controls (n=12) were investigated for the presence of antibodies that reacted with unstimulated and cytokine stimulated BECs isolated from a normal healthy liver. To demonstrate organ specificity, lung epithelial cells (LECs) were used as control cells. Antibodies were tested for their functional capacity. RESULTS: Compared with controls (8%), significantly higher numbers of PSC patients (63%, p=0.001), but not PBC (37%, NS) or AIH (16%, NS) patients, had anti-BEC antibodies. In 90% of PSC patients, the autoantibodies reacted only with cytokine stimulated target cells. Lower numbers of PSC (6%), PBC (10%), and AIH (0%) patients had LEC antibodies. Other significant findings were that anti-BEC antibodies were found in (i) PSC patients with either the HLA-DRB1*0301 or DR2 allele compared with those without (p=0.007); and (ii) in PBC patients with end stage disease compared with those without (p=0.018). Furthermore, anti-BEC antibodies from PSC and PBC but not AIH patients induced BECs to produce high levels of the cytokine interleukin 6. IgM and IgG fractions isolated from PSC but not PBC and AIH sera induced significantly increased expression of the cell adhesion molecule CD44. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and western blot analysis of BEC membranes demonstrated a specific band of 40 kDa with PSC sera and 45, 42, 30, and 33 kDa bands with PBC sera, which were absent in control groups. CONCLUSION: Thus for the first time we have demonstrated the presence of functionally important autoantibodies to cell surface expressed antigens on the relevant target cells of destruction, namely BECs, in PSC and PBC. These finding have important implications for the pathogenesis of bile duct destruction in these patients. PMID- 12077103 TI - Hepatoprotection with tauroursodeoxycholate and beta muricholate against taurolithocholate induced cholestasis: involvement of signal transduction pathways. AB - BACKGROUND: Tauroursodeoxycholate (TUDC) provides partial protection against taurolithocholate (TLC) induced cholestasis, possibly by inducing a signalling cascade activating protein kinase C (PKC). The potential protective effects of beta muricholic acid (beta-MC), another 7-beta-hydroxylated bile salt, have not previously been studied in TLC cholestasis. AIMS: To study the effect of beta-MC on TLC induced cholestasis and also to investigate further the effects of agents affecting intracellular signalling, notably DBcAMP (a cell permeable cAMP analogue) and several protein kinase inhibitors. METHODS: Functional studies were carried out analysing the proportion of hepatocyte couplets able to accumulate the fluorescent bile acid analogue cholyl-lysyl-fluorescein (CLF) into their sealed canalicular vacuole (cVA of CLF assay). RESULTS: It was found that both beta-MC and DBcAMP were as effective as TUDC in protecting against TLC induced cholestasis. The PKC inhibitors staurosporin and H7 but not the specific protein kinase A (PKA) inhibitor KT5720 abolished the protective effects of TUDC and beta MC. BAPTA/AM, a chelator of intracellular Ca(2+), significantly decreased the protective effect of both bile salts, and that of DBcAMP. PKC and PKA inhibitors had no effect on protection with DBcAMP. CONCLUSIONS: Beta-MC was as effective as TUDC in protecting against TLC cholestasis. Mobilisation of Ca(2+) and activation of PKC, but not of PKA, are involved in the anticholestatic effect of the two 7 beta-hydroxylated bile salts. The hepatoprotective effects of DBcAMP involved Ca(2+) mobilisation, but not PKC or PKA activation. PMID- 12077105 TI - Cholangiocarcinoma arising after biliary-enteric drainage procedures for benign disease. PMID- 12077106 TI - Gastrointestinal epithelial neoplasia: Vienna revisited. AB - International consensus meetings in Padova and Vienna have attempted to rationalise the grading and classification of gastrointestinal epithelial neoplasia (GEN). With its minor adjustments, the Vienna classification of GEN seeks to be more closely in tune with patient management and it is hoped that it is not seen as fiddling around with terms but as a genuine contribution to patient care. PMID- 12077108 TI - Upper gastrointestinal bleeding and surrogate end points. PMID- 12077109 TI - Gluten exposure and risk of autoimmune disorders. PMID- 12077107 TI - Cocarcinogenic effects of alcohol in hepatocarcinogenesis. AB - Alcohol is a major aetiological factor in hepatocarcinogenesis but our understanding of its importance as a modulating factor is just beginning to emerge. In the present review, a number of possible cofactors and mechanisms are discussed by which alcohol may enhance the development of hepatoma. These include dietary or environmental carcinogens ingested along with alcoholic beverages, alcoholic cirrhosis as a precancerous condition, and the effects of alcohol metabolism. PMID- 12077110 TI - Thiopurine metabolites and the role of thiopurine methyltransferase activity in inflammatory bowel disease. PMID- 12077111 TI - Serum leptin and body mass index in children with H pylori infection. PMID- 12077112 TI - Inhibition of nucleoside transport by p38 MAPK inhibitors. AB - While investigating the ability of p38 MAPK to regulate cytarabine (Ara C) dependent differentiation of erythroleukemia K562 cells, we observed effects that indicated that the imidazoline class of p38 MAPK inhibitors prevented nucleoside transport. Incubation of K562 cells with SB203580, SB203580-iodo, or SB202474, an analogue of SB203580 that does not inhibit p38 MAPK activity, inhibited the uptake of [3H]Ara C or [3H]uridine and the differentiation of K562 cells. Consistent with the effects of these compounds on the nitrobenzylthioinosine (NBMPR)-sensitive equilibrative nucleoside transporter (ENT1), incubation with SB203580 or SB203580-iodo eliminated the binding of [3H]NBMPR to K562 cells or membranes isolated from human erythrocytes. Furthermore, using a uridine dependent cell type (G9c), we observed that SB203580 or SB203580-iodo efficiently inhibited the salvage synthesis of pyrimidine nucleotides in vivo. Thus these studies demonstrate that the NBMPR-sensitive equilibrative nucleoside transporters are novel and unexpected targets for the p38 MAPK inhibitors at concentrations typically used to inhibit protein kinases. PMID- 12077113 TI - Wnt-3A enhances bone morphogenetic protein-2-mediated chondrogenesis of murine C3H10T1/2 mesenchymal cells. AB - We have recently reported the chondrogenic effect of bone morphogenetic protein-2 (BMP-2) in high density cultures of the mouse multipotent mesenchymal C3H10T1/2 cell line and have shown the functional requirement of the cell-cell adhesion molecule N-cadherin in BMP-2-induced chondrogenesis in vitro (Denker, A. E., Nicoll, S. B., and Tuan, R. S. (1995) Differentiation 59, 25-34; Haas, A. R., and Tuan, R. S. (1999) Differentiation 64, 77-89). Furthermore, BMP-2 treatment also results in an increased protein level of beta-catenin, a known N-cadherin associated Wnt signal transducer (Fischer, L., Haas, A., and Tuan, R. S. (2001) Signal Transduction 2, 66-78), suggesting functional cross-talk between the BMP-2 and Wnt signaling pathways. We have observed previously that BMP-2 treatment up regulates expression of Wnt-3A in high density cultures of C3H10T1/2 cells. To assess the contribution of Wnt-3A to BMP-2-mediated chondrogenesis, we have generated C3H10T1/2 cell lines overexpressing Wnt-3A and various forms of glycogen synthase kinase-3beta (GSK-3beta), an immediate cytosolic component of the Wnt signaling pathway, and examined their response to BMP-2. We show that overexpression of either Wnt-3A or kinase-dead GSK-3beta enhances BMP-2-mediated chondrogenesis. Furthermore, Wnt-3A overexpression results in decreases in both N cadherin and GSK-3beta protein levels, whereas Wnt-3A as well as kinase-dead GSK 3beta overexpression increase total and nuclear levels of both beta-catenin and LEF-1. Direct cross-talk between Wnts and BMP-2 was also indicated by the up regulated interaction between beta-catenin and SMAD-4 in response to BMP-2. These results suggest that Wnt-3A acts in a manner opposite to that of other Wnts, such as Wnt-7A, which were previously identified as inhibitory to chondrogenesis, and is the first BMP-2-regulated, chondrogenesis-enhancing member of the Wnt family. PMID- 12077114 TI - Analysis of the structural basis of specificity of inhibition of the Abl kinase by STI571. AB - STI571, a selective inhibitor of Bcr-Abl, has been a successful therapeutic agent in clinical trials for chronic myelogenous leukemia. Chronic phase chronic myelogenous leukemia patients treated with STI571 have durable responses; however, most responding blast phase patients relapse despite continued therapy. Co-crystallization studies of Abl kinase and an STI571-related compound identify specific amino acid residues as critical to STI571 binding, one of which, T315, has been characterized as an acquired Thr to Ile mutation in relapsed patients. Other studies, however, suggest that mutations other than these predicted contact points are capable of conferring STI571 resistance in relapsed patients. Using a variety of models of STI571 binding to the Abl kinase, we have performed an extensive mutational analysis of sites that might alter the sensitivity of the Abl kinase to STI571. Although mutation of many of the predicted contact points between Abl and STI571 result in a kinase-inactive protein, additional mutations that render the Abl kinase less sensitive to STI571 demonstrate a broad range of possibilities for clinical resistance that are now becoming evident. PMID- 12077115 TI - Induction of protein conformational change in mouse senile amyloidosis. AB - Aggregated amyloid fibrils can induce further polymerization of precursor proteins in vitro, thus providing a possible basis for propagation or transmission in the pathogenesis of amyloidoses. Previously, we postulated that the transmission of amyloid fibrils induces conformational changes of endogenous amyloid protein in mouse senile amyloidosis (Xing, Y., Nakamura, A., Chiba, T., Kogishi, K., Matsushita, T., Fu, L., Guo Z., Hosokawa, M., Mori, M., and Higuchi, K. (2001) Lab. Invest. 81, 493-499). To further characterize this transmissibility, we injected amyloid fibrils (AApoAII(C)) of amyloidogenic C type apolipoprotein A-II (APOAIIC) intravenously into 2-month-old SAMR1 mice, which have B type apolipoprotein A-II (APOAIIB), and develop few if any amyloid deposits spontaneously. 10 months after amyloid injection, deposits were detected in the tongue, stomach, intestine, lungs, heart, liver, and kidneys. The intensity of deposition increased thereafter, whereas no amyloid was detected in distilled water-injected SAMR1 mice, even after 20 months. The deposited amyloid was composed of endogenous APOAIIB with a different amyloid fibril conformation. The injection of these amyloid fibrils of APOAIIB (AApoAII(B)) induced earlier and more severe amyloidosis in SAMR1 mice than the injection of AApoAII(C) amyloid fibrils. Thus, AApoAII(C) from amyloidogenic mice could induce a conformational change of less amyloidogenic APOAIIB to a different amyloid fibril structure, which could also induce amyloidosis in the less amyloidogenic strain. These results provide important insights into the pathogenesis of amyloid diseases. PMID- 12077116 TI - Mitochondrial targeted cyclophilin D protects cells from cell death by peptidyl prolyl isomerization. AB - Cyclophilin D (CyPD) is thought to sensitize opening of the mitochondrial permeability transition pore (mPTP) based on the findings that cyclosporin A (CsA), a pseudo-CyPD substrate, hyperpolarizes the mitochondrial membrane potential (DeltaPsi) and inhibits apoptosis. We provide evidence that contrasts with this model. Using live cell imaging and two photon microscopy, we report that overexpression of CyPD desensitizes HEK293 and rat glioma C6 cells to apoptotic stimuli. By site-directed mutagenesis of CyPD that compromises peptidyl prolyl cis-trans isomerase (PPIase) activity, we demonstrate that the mechanism involved in this protective effect requires PPIase activity. Furthermore, we show that, under resting conditions, DeltaPsi is hyperpolarized in CyPD wild type overexpressing cells but not in cells overexpressing mutant forms of CyPD that lack PPIase activity. Finally, in glutathione S-transferase (GST) pull-down assays, we demonstrate that CyPD binding to the adenine nucleotide translocator (ANT), which is considered to be the core component of the mPTP, is not affected by the loss of PPIase activity. Collectively, our data suggest that CyPD should be viewed as a cell survival-signaling molecule and indicate a protective role of CyPD against apoptosis that is mediated by one or more targets other than the ANT. PMID- 12077117 TI - 85-kDa cytosolic phospholipase A2 mediates peroxisome proliferator-activated receptor gamma activation in human lung epithelial cells. AB - The 85-kDa cytosolic phospholipase A(2) (cPLA(2)) plays an important role in the control of arachidonic acid metabolism. This study was designed to investigate the possible contributions of cPLA(2) and group IIA secretory phospholipase A(2) (sPLA(2)) in the regulation of peroxisome proliferator-activated receptor (PPAR) mediated gene transcription in human airway epithelial cells. Primary normal human bronchial epithelial cells and human lung epithelial cell lines BEAS 2B, A549, and NCI-H292 all express PPARgamma and -beta. Overexpression of cPLA(2) in BEAS 2B cells and primary bronchial epithelial cells resulted in a significant increase of PPARgamma-mediated reporter activity. In contrast, overexpression of group IIA sPLA(2) had no effect on PPARgamma activation. The PPARgamma activity in A549 cells was significantly inhibited by the cPLA(2) inhibitor arachidonyltrifluoromethyl ketone but not by the sPLA(2) inhibitor LY311727 and the iPLA(2) inhibitor HELSS. Activation of cPLA(2) by the calcium ionophore, induced a dose-dependent increase of PPAR activity in normal human bronchial epithelial cells and in the A549 cells. Electrophoretic mobility shift assays show that the binding between PPAR isolated from A549 cells and peroxisome proliferator response element (PPRE) is enhanced by but partially blocked by the cPLA(2) inhibitors arachidonyltrifluoromethyl ketone and methyl arachidonyl fluorophosphate. Finally, NS 398, a COX-2 inhibitor, partially blocked the effect on PPAR activity and binding to the PPRE suggesting involvement of COX-2 metabolites in PPRE activation. The above results demonstrate a novel function of cPLA(2) in the control of PPARgamma activation in human lung epithelial cells. PMID- 12077118 TI - Phorbol 12-myristate 13-acetate up-regulates the transcription of MUC2 intestinal mucin via Ras, ERK, and NF-kappa B. AB - MUC2 is a secretory mucin normally expressed by goblet cells of the intestinal epithelium. It is overexpressed in mucinous type colorectal cancers but down regulated in colorectal adenocarcinoma. Phorbol 12-myristate 13-acetate (PMA) treatment of colon cancer cell lines increases MUC2 expression, so we have undertaken a detailed analysis of the effects of PMA on the promoter activity of the 5'-flanking region of the MUC2 gene using stably and transiently transfected promoter reporter vectors. Protein kinase C inhibitors (bisindolylmaleimide, calphostin C) and inhibitors of mitogen-activated protein/extracellular signal regulated kinase kinase (MEK) (PD98059 and U0126) suppressed up-regulation of MUC2. Src tyrosine kinase inhibitor PP2, a protein kinase A inhibitor (KT5720), and a p38 inhibitor (SB 203580) did not affect transcription. Western blotting and reverse transcription-PCR analysis confirmed these results. In addition, co transfections with mutants of Ras, Raf, and MEK showed that the induction of MUC2 promoter activity by PMA required these three signaling proteins. Our results demonstrate that PMA activates protein kinase C, stimulating MAP kinase through a Ras- and Raf-dependent mechanism. An important role for nuclear factor kappaB (NF kappaB) was also demonstrated using the inhibitor caffeic acid phenethyl ester and electrophoretic mobility shift assays. Such identification of pathways involved in MUC2 up-regulation by PMA in the HM3 colon cancer cell line may serve as a model for the effects of cytokines and growth factors, which regulate MUC2 expression during the progression of colorectal cancer. PMID- 12077119 TI - DNA template requirements for human mismatch repair in vitro. AB - The human mismatch repair pathway is competent to correct DNA mismatches in a strand-specific manner. At present, only nicks are known to support strand discrimination, although the DNA end within the active site of replication is often proposed to serve this role. We therefore tested the competence of DNA ends or gaps to direct mismatch correction. Eight G.T templates were constructed which contained a nick or gap of 4, 28, or approximately 200 nucleotides situated approximately 330 bp away in either orientation. A competition was established in which the mismatch repair machinery had to compete with gap-filling replication and ligation activities for access to the strand discontinuity. Gaps of 4 or 28 nucleotides were the most effective strand discrimination signals for mismatch repair, whereas double strand breaks did not direct repair to either strand. To define the minimal spatial requirements for access to either the strand signal or mismatch site, the nicked templates were linearized close to either site and assayed. As few as 14 bp beyond the nick supported mismatch excision, although repair synthesis failed using 5'-nicked templates. Finally, asymmetric G.T templates with a remote nick and a nearby DNA end were repaired efficiently. PMID- 12077120 TI - Interaction of 14-3-3 protein with regulator of G protein signaling 7 is dynamically regulated by tumor necrosis factor-alpha. AB - Regulators of G protein signaling (RGS) constitute a family of proteins with a conserved RGS domain of approximately 120 amino acids that accelerate the intrinsic GTP hydrolysis of activated Galpha(i) and Galpha(q) subunits. The phosphorylation-dependent interaction of 14-3-3 proteins with a subset of RGS proteins inhibits their GTPase-accelerating activity in vitro. The inhibitory interaction between 14-3-3 and RGS7 requires phosphorylation of serine 434 of RGS7. We now show that phosphorylation of serine 434 is dynamically regulated by TNF-alpha. Cellular stimulation by TNF-alpha transiently decreased the phosphorylation of serine 434 of RGS7, abrogating the inhibitory interaction with 14-3-3. We examined the effect of 14-3-3 on RGS-mediated deactivation kinetics of G protein-coupled inwardly rectifying K(+) channels (GIRKs) in Xenopus oocytes. 14-3-3 inhibited the function of wild-type RGS7, but not that of either RSG7(P436R) or RGS4, two proteins that do not bind 14-3-3. Our findings are the first evidence that extracellular signals can modulate the activity of RGS proteins by regulating their interaction with 14-3-3. PMID- 12077121 TI - YOS9, the putative yeast homolog of a gene amplified in osteosarcomas, is involved in the endoplasmic reticulum (ER)-Golgi transport of GPI-anchored proteins. AB - The OS-9 gene maps to a region (q13-15) of chromosome 12 that is highly amplified in human osteosarcomas and encodes a protein of unknown function. Here we have characterized a homolog designated as YOS9 (YDR057w) from Saccharomyces cerevisiae. The yeast protein (Yos9) is a membrane-associated glycoprotein that localizes to the endoplasmic reticulum (ER). YOS9 interacts genetically with genes involved in ER-Golgi transport, particularly SEC34, whose temperature sensitive mutant is rescued by YOS9 overexpression. Interestingly, Yos9 appears to play a direct role in the transport of glycosylphosphatidylinositol (GPI) anchored proteins to the Golgi apparatus. Yos9 binds directly to Gas1 and Mkc7 and accelerates Gas1 transport and processing in cells overexpressing YOS9. Correspondingly, Gas1 processing is slowed in cells bearing a deletion in YOS9. No effect upon the transport and processing of non-GPI-anchored proteins (e.g. invertase and carboxypeptidase Y) was detected in cells either lacking or overexpressing Yos9. As Yos9 is not a component of the Emp24 complex, it may act as a novel escort factor for GPI-anchored proteins in ER-Golgi transport in yeast and possibly in mammals. PMID- 12077122 TI - Regulation of signaling in B cells through the phosphorylation of Syk on linker region tyrosines. A mechanism for negative signaling by the Lyn tyrosine kinase. AB - The B cell antigen receptor (BCR) is coupled to the mobilization of Ca(2+) by the protein-tyrosine kinase, Syk. Syk, recruited to the clustered BCR, becomes phosphorylated on three tyrosines (Tyr-317, Tyr-342, and Tyr-346) located within the linker region that separates the C-terminal catalytic domain from the N terminal tandem Src homology 2 domains. Phosphorylation within the linker region can be either activating or inhibitory to Ca(2+) mobilization depending on the sites that are modified. Syk that is not phosphorylated on linker region tyrosines couples the BCR to Ca(2+) mobilization through a phosphoinositide 3 kinase-dependent pathway. The phosphorylation of Tyr-342 and -346 enhances the phosphorylation and activation of phospholipase C-gamma and the early phase of Ca(2+) mobilization via a phosphoinositide 3-kinase-independent pathway. The phosphorylation of Tyr-317 strongly dampens the Ca(2+) signal. In cells that lack the Src family kinase, Lyn, the phosphorylation of the inhibitory Tyr-317 is suppressed leading to elevated production of inositol 1,4,5-trisphosphate and an amplified Ca(2+) signal. This provides a novel mechanism by which Lyn functions as an inhibitor of BCR-stimulated signaling. Thus, Syk and Lyn combine to determine the pathway through which the BCR is coupled to Ca(2+) mobilization as well as the magnitude and duration of the Ca(2+) flux. PMID- 12077123 TI - Identification and characterization of RPK118, a novel sphingosine kinase-1 binding protein. AB - Sphingosine kinase (SPHK) is a key enzyme catalyzing the formation of sphingosine 1 phosphate (SPP), a lipid messenger that is implicated in the regulation of a wide variety of important cellular events through intracellular as well as extracellular mechanisms. However, the molecular mechanism of the intracellular actions of SPP remains unclear. Here we have cloned a novel sphingosine kinase-1 (SPHK1)-binding protein, RPK118, by yeast two-hybrid screening. RPK118 contains several functional domains whose sequences are homologous to other known proteins including the phox homology domain and pseudokinase 1 and 2 domains and is shown to be a member of an evolutionarily highly conserved gene family. The pseudokinase 2 domain of RPK118 is responsible for SPHK1 binding as judged by yeast two-hybrid screening and immunoprecipitation studies. RPK118 is also shown to co-localize with SPHK1 on early endosomes in COS7 cells expressing both recombinant proteins. Furthermore, RPK118 specifically binds to phosphatidylinositol 3-phosphate. These results strongly suggest that RPK118 is a novel SPHK1-binding protein that may be involved in transmitting SPP-mediated signaling into the cell. PMID- 12077124 TI - Metabolism of 4 beta -hydroxycholesterol in humans. AB - One of the major oxysterols in the human circulation is 4 beta-hydroxycholesterol formed from cholesterol by the drug-metabolizing enzyme cytochrome P450 3A4. Deuterium-labeled 4 beta-hydroxycholesterol was injected into two healthy volunteers, and the apparent half-life was found to be 64 and 60 h, respectively. We have determined earlier the half-lives for 7 alpha-, 27-, and 24 hydroxycholesterol to be approximately 0.5, 0.75, and 14 h, respectively. Patients treated with certain antiepileptic drugs have up to 20-fold increased plasma concentrations of 4 beta-hydroxycholesterol. The apparent half-life of deuterium-labeled 4 beta-hydroxycholesterol in such a patient was found to be 52 h, suggesting that the high plasma concentration was because of increased synthesis rather than impaired clearance. 4 beta-Hydroxycholesterol was converted into acidic products at a much slower rate than 7 alpha-hydroxycholesterol in primary human hepatocytes, and 4 beta-hydroxycholesterol was 7 alpha-hydroxylated at a slower rate than cholesterol by recombinant human CYP7A1. CYP7B1 and CYP39A1 had no activity toward 4 beta-hydroxycholesterol. These results suggest that the high plasma concentration of 4 beta-hydroxycholesterol is because of its exceptionally slow elimination, probably in part because of the low rate of 7 alpha-hydroxylation of the steroid. The findings are discussed in relation to a potential role of 4 beta-hydroxycholesterol as a ligand for the nuclear receptor LXR. PMID- 12077125 TI - Structures of CUG repeats in RNA. Potential implications for human genetic diseases. AB - Triplet repeats that cause human genetic diseases have been shown to exhibit unusual compact structures in DNA, and in this paper we show that similar structures exist in shorter "normal length" CNG RNA. CUG and control RNAs were made chemically and by in vitro transcription. We find that "normal" short CUG RNAs migrate anomalously fast on non-denaturing gels, compared with control oligos of similar base composition. By contrast, longer tracts approaching clinically relevant lengths appear to form higher order structures. The CD spectrum of shorter tracts is similar to triplex and pseudoknot nucleic acid structures and different from classical hairpin spectra. A model is outlined that enables the base stacking features of poly(r(G-C))(2).poly(r(U)) or poly(d(G C))(2).poly(d(T)) triplexes to be achieved, even by a single 15-mer. PMID- 12077126 TI - Electrostatic control of the isoalloxazine environment in the two-electron reduced states of yeast glutathione reductase. AB - The resonance Raman spectra of the oxidized and two-electron reduced forms of yeast glutathione reductase are reported. The spectra of the oxidized enzyme indicate a low electron density for the isoalloxazine ring. As far as the two electron reduced species are concerned, the spectral comparison of the NADPH reduced enzyme with the glutathione- or dithiothreitol-reduced enzyme shows significant frequency differences for the flavin bands II, III, and VII. The shift of band VII was correlated with a change in steric or electronic interaction of the hydroxyl group of a conserved Tyr with the N(10)-C(10a) portion of the isoalloxazine ring. Upward shifts of bands II and III observed for the glutathione- or dithiothreitol-reduced enzyme indicate both a slight change in isoalloxazine conformation and a hydrogen bond strengthening at the N(1) and/or N(5) site(s). The formation of a mixed disulfide intermediate tends to slightly decrease the frequency of bands II, III, X, XI, and XIV. To account for the different spectral features observed for the NADPH- and glutathione-reduced species, several possibilities have been examined. In particular, we propose a hydrogen bonding modulation at the N(5) site of FAD through a variable conformation of an ammonium group of a conserved Lys residue. Changes in N(5)(flavin)-protein interaction in the two-electron reduced forms of glutathione reductase are discussed in relation to a plausible mechanism of the regulation of the enzyme activity via a variable redox potential of FAD. PMID- 12077127 TI - Fast and slow inactivation kinetics of the Ca2+ channels ECaC1 and ECaC2 (TRPV5 and TRPV6). Role of the intracellular loop located between transmembrane segments 2 and 3. AB - The Ca(2+) channels ECaC1 and ECaC2 (TRPV5 and TRPV6) share several functional properties including permeation profile and Ca(2+)-dependent inactivation. However, the kinetics of ECaC2 currents notably differ from ECaC1 currents. The initial inactivation is much faster in ECaC2 than in ECaC1, and the kinetic differences between Ca(2+) and Ba(2+) currents are more pronounced for ECaC2 than ECaC1. Here, we identify the structural determinants for these functional differences. Chimeric proteins were expressed heterologously in HEK 293 cells and studied by patch clamp analysis. Both channels retained their phenotype after exchanging the complete N termini, the C termini, or even both N and C termini, i.e. ECaC1 with the ECaC2 N or C terminus still showed the ECaC1 phenotype and vice versa. The substitution of the intracellular loop between the transmembrane domains 2 and 3 of ECaC2 with that of ECaC1 induced a delay of inactivation. Three amino acid residues (Leu-409, Val-411 and Thr-412) present in this loop determine the fast inactivation behavior. When this intracellular loop between the transmembrane domains 2 and 3 of ECaC1 was exchanged with the TM2-TM3 loop of ECaC2, the ECaC1 kinetics were analogous to ECaC2. In conclusion, the TM2-TM3 loop is a critical determinant of the inactivation in ECaC1 and ECaC2. PMID- 12077128 TI - Threonine 308 within a putative casein kinase 2 site of the cytoplasmic tail of leukotriene B(4) receptor (BLT1) is crucial for ligand-induced, G-protein-coupled receptor-specific kinase 6-mediated desensitization. AB - Desensitization of G-protein-coupled receptors may involve phosphorylation of serine and threonine residues. The leukotriene B(4) (LTB(4)) receptor (BLT1) contains 14 intracellular serines and threonines, 8 of which are part of consensus target sequences for protein kinase C (PKC) or casein kinase 2. In this study, we investigated the importance of PKC and GPCR-specific kinase (GRK) phosphorylation in BLT1 desensitization. Pretreatment of BLT1-transfected COS-7 cells with PKC activators caused a decrease of LTB(4)-induced inositol phosphate (IP) accumulation. This reduction was prevented with the PKC inhibitor, staurosporine, and not observed in cells expressing a BLT1 deletion mutant (G291stop) lacking the cytoplasmic tail. Moreover LTB(4)-induced IP accumulation was significantly inhibited by overexpression of GRK2, GRK5, and especially GRK6, in cells expressing wild type BLT1 but not in those expressing G291stop. GRK6 mediated desensitization correlated with increased phosphorylation of BLT1. The G319stop truncated BLT1 mutant displayed functional characteristics comparable with wild type BLT1 in terms of desensitization by GRK6, but not by PKC. Substitution of Thr(308) within a putative casein kinase 2 site to proline or alanine in the full-length BLT1 receptor prevented most of GRK6-mediated inhibition of LTB(4)-induced IP production but only partially affected LTB(4) induced BLT1 phosphorylation. Our findings thus suggest that Thr(308) is a major residue involved in GRK6-mediated desensitization of BLT1 signaling. PMID- 12077129 TI - Functional divergence in the glutathione transferase superfamily in plants. Identification of two classes with putative functions in redox homeostasis in Arabidopsis thaliana. AB - Searches with the human Omega glutathione transferase (GST) identified two outlying groups of the GST superfamily in Arabidopsis thaliana which differed from all other plant GSTs by containing a cysteine in place of a serine at the active site. One group consisted of four genes, three of which encoded active glutathione-dependent dehydroascorbate reductases (DHARs). Two DHARs were predicted to be cytosolic, whereas the other contained a chloroplast targeting peptide. The DHARs were also active as thiol transferases but had no glutathione conjugating activity. Unlike most other GSTs, DHARs were monomeric. The other class of GST comprised two genes termed the Lambda GSTs (GSTLs). The recombinant GSTLs were also monomeric and had glutathione-dependent thiol transferase activity. One GSTL was cytosolic, whereas the other was chloroplast-targeted. When incubated with oxidized glutathione, the putative active site cysteine of the GSTLs and cytosolic DHARs formed mixed disulfides with glutathione, whereas the plastidic DHAR formed an intramolecular disulfide. DHAR S-glutathionylation was consistent with a proposed catalytic mechanism for dehydroascorbate reduction. Roles for the cytosolic DHARs and GSTLs as antioxidant enzymes were also inferred from the induction of the respective genes following exposure to chemicals and oxidative stress. PMID- 12077130 TI - Characterization of heparanase from a rat parathyroid cell line. AB - Cell surface heparan sulfate proteoglycans undergo unique intracellular degradation pathways after they are endocytosed from the cell surface. Heparanase, an endo-beta-glucuronidase capable of cleaving heparan sulfate, has been demonstrated to contribute to the physiological degradation of heparan sulfate proteoglycans and therefore regulation of their biological functions. A rat parathyroid cell line was found to produce heparanase with an optimal activity at neutral and slightly acidic conditions suggesting that the enzyme participates in heparan sulfate proteoglycan metabolism in extralysosomal compartments. To elucidate the detailed properties of the purified enzyme, the substrate specificity against naturally occurring heparan sulfates and chemically modified heparins was studied. Cleavage sites of rat heparanase were present in heparan sulfate chains obtained from a variety of animal organs, but their occurrence was infrequent (average, 1-2 sites per chain) requiring recognition of both undersulfated and sulfated regions of heparan sulfate. On the other hand intact and chemically modified heparins were not cleaved by heparanase. The carbohydrate structure of the newly generated reducing end region of heparan sulfate cleaved by the enzyme was determined, and it represented relatively undersulfated structures. O-Sulfation of heparan sulfate chains also played important roles in substrate recognition, implying that rat parathyroid heparanase acts near the boundary of highly sulfated and undersulfated domains of heparan sulfate proteoglycans. Further elucidation of the roles of heparanase in normal physiological processes would provide an important tool for analyzing the regulation of heparan sulfate-dependent cell functions. PMID- 12077131 TI - Tumor necrosis factor induces apoptosis in hepatoma cells by increasing Ca(2+) release from the endoplasmic reticulum and suppressing Bcl-2 expression. AB - Tumor necrosis factor (TNF) plays an import role in the control of apoptosis. The most well known apoptotic pathway regulated by TNF involves the TNFR1-associated death domain protein, Fas-associated death domain protein, and caspase-8. This study examines the mechanism of TNF-induced apoptosis in FaO rat hepatoma cells. TNF treatment significantly increased the percentage of apoptotic cells. TNF did not activate caspase-8 but activated caspase-3, -10, and -12. The effect of TNF on the expression of different members of the Bcl-2 family in these cells was studied. We observed no detectable changes in the steady-state levels of Bcl X(L), Bax, and Bid, although TNF suppresses Bcl-2 expression. Dantrolene suppressed the inhibitory effect of TNF on Bcl-2 expression. TNF induced release of Ca(2+) from the endoplasmic reticulum (ER) that was blocked by dantrolene. Importantly, the expression of Bcl-2 blocked TNF-induced apoptosis and decreased TNF-induced Ca(2+) release. These results suggest that TNF induces apoptosis by a mechanism that involves increasing Ca(2+) release from the ER and suppression of Bcl-2 expression. PMID- 12077132 TI - Integrins alpha(6A)beta 1 and alpha(6B)beta 1 promote different stages of chondrogenic cell differentiation. AB - The differentiation of chondrocytes and of several other cell types is associated with a switch from the alpha(6B) to the alpha(6A) isoform of the laminin alpha(6)beta(1) integrin receptor. To define whether this event plays a functional role in cell differentiation, we used an in vitro model system that allows chick chondrogenic cells to remain undifferentiated when cultured in monolayer and to differentiate into chondrocytes when grown in suspension culture. We report that: (i) upon over-expression of the human alpha(6B), adherent chondrogenic cells differentiate to stage I chondrocytes (i.e. increased type II collagen, reduced type I collagen, fibronectin, alpha(5)beta(1) and growth rate, loss of fibroblast morphology); (ii) the expression of type II collagen requires the activation of p38 MAP kinase; (iii) the over-expression of alpha(6A) induces an incomplete differentiation to stage I chondrocytes, whereas no differentiation was observed in alpha(5) and mock-transfected control cells; (iv) a prevalence of the alpha(6A) subunit is necessary to stabilize the differentiated phenotype when cells are transferred to suspension culture. Altogether, these results indicate a functional role for the alpha(6B) to alpha(6A) switch in chondrocyte differentiation; the former promotes chondrocyte differentiation, and the latter is necessary in stabilizing the differentiated phenotype. PMID- 12077133 TI - Rad52 protein associates with replication protein A (RPA)-single-stranded DNA to accelerate Rad51-mediated displacement of RPA and presynaptic complex formation. AB - The Rad51 nucleoprotein filament mediates DNA strand exchange, a key step of homologous recombination. This activity is stimulated by replication protein A (RPA), but only when RPA is introduced after Rad51 nucleoprotein filament formation. In contrast, RPA inhibits Rad51 nucleoprotein complex formation by prior binding to single-stranded DNA (ssDNA), but Rad52 protein alleviates this inhibition. Here we show that Rad51 filament formation is simultaneous with displacement of RPA from ssDNA. This displacement is initiated by a rate-limiting nucleation of Rad51 protein onto ssDNA complex, followed by rapid elongation of the filament. Rad52 protein accelerates RPA displacement by Rad51 protein. This acceleration probably involves direct interactions with both Rad51 protein and RPA. Detection of a Rad52-RPA-ssDNA co-complex suggests that this co-complex is an intermediate in the displacement process. PMID- 12077134 TI - Activation of the p38 signaling pathway by heat shock involves the dissociation of glutathione S-transferase Mu from Ask1. AB - Despite the importance of the stress-activated protein kinase pathways in cell death and survival, it is unclear how stressful stimuli lead to their activation. In the case of heat shock, the existence of a specific mechanism of activation has been evidenced, but the molecular nature of this pathway is undefined. Here, we found that Ask1 (apoptosis signal-regulating kinase 1), an upstream activator of the stress-activated protein kinase p38 during exposure to oxidative stress and other stressful stimuli, was also activated by heat shock. Ask1 activity was required for p38 activation since overexpression of a kinase dead mutant of Ask1, Ask1(K709M), inhibited heat shock-induced p38 activation. The activation of Ask1 by oxidative stress involves the oxidation of thioredoxin, an endogenous inhibitor of Ask1. A different activation mechanism takes place during heat shock. In contrast to p38 induction by H(2)O(2), induction by heat shock was not antagonized by pretreatment with the antioxidant N-acetyl-l-cysteine or by overexpressing thioredoxin and was not accompanied by the dissociation of thioredoxin from Ask1. Instead, heat shock caused the dissociation of glutathione S-transferase Mu1-1 (GSTM1-1) from Ask1 and overexpression of GSTM1-1-inhibited induction of p38 by heat shock. We concluded that because of an alternative regulation by the two distinct repressors thioredoxin and GSTM1-1, Ask1 constitutes the converging point of the heat shock and oxidative stress-sensing pathways that lead to p38 activation. PMID- 12077135 TI - Comparative trajectories of active and S195A inactive trypsin upon binding to serpins. AB - Serpins inhibit proteinases through a complicated multistep mechanism. The precise nature of these steps and the order by which they occur are still debated. We compared the fate of active and S195A inactive rat trypsin upon binding to alpha(1)-antitrypsin and P(1)-Arg-antichymotrypsin using stopped-flow kinetics with fluorescence resonance energy transfer detection and time-resolved fluorescence resonance energy transfer. We show that inhibition of active trypsin by these serpins leads to two irreversible complexes, one being compatible with the full insertion of the serpin-reactive site loop but not the other one. Binding of inactive trypsin to serpins triggers a large multistep reversible rearrangement leading to the migration of the proteinase to an intermediate position. Binding of inactive trypsin, unlike that of active trypsin, does not perturb the rhodamine fluorescence at position 150 on the helix F of the serpin. Thus, inactive proteinases do not migrate past helix F and do not trigger full serpin loop insertion. PMID- 12077136 TI - The linker region plays an important role in the interdomain communication of the response regulator OmpR. AB - OmpR is the response regulator of a two-component regulatory system that controls the expression of the porin genes ompF and ompC in Escherichia coli. This regulator consists of two domains joined by a flexible linker region. The amino terminal domain is phosphorylated by the sensor kinase EnvZ, and the carboxyl terminal domain binds DNA via a winged helix-turn-helix motif. In vitro studies have shown that amino-terminal phosphorylation enhances the DNA binding affinity of OmpR and, conversely, that DNA binding by the carboxyl terminus increases OmpR phosphorylation. In the present work, we demonstrate that the linker region contributes to this communication between the two domains of OmpR. Changing the specific amino acid composition of the linker alters OmpR function, as does increasing or decreasing its length. Three linker mutants give rise to an OmpF(+) OmpC(-) phenotype, but the defects are not due to a shared molecular mechanism. Currently, functional homology between response regulators is predicted based on similarities in the amino and carboxyl-terminal domains. The results presented here indicate that linker length and composition should also be considered. Furthermore, classification of response regulators in the same subfamily does not necessarily imply that they share a common response mechanism. PMID- 12077137 TI - The cryptic adenine deaminase gene of Escherichia coli. Silencing by the nucleoid associated DNA-binding protein, H-NS, and activation by insertion elements. AB - In Escherichia coli there are two pathways for conversion of adenine into guanine nucleotides, both involving the intermediary formation of IMP. The major pathway involves conversion of adenine into hypoxanthine in three steps via adenosine and inosine, with subsequent phosphoribosylation of hypoxanthine to IMP. The minor pathway involves formation of ATP, which is converted via the histidine pathway to the purine intermediate 5-amino-4-imidazolecarboxamide ribonucleotide and, subsequently, to IMP. Here we describe E. coli mutants, in which a third pathway for conversion of adenine to IMP has been activated. This pathway was shown to involve direct deamination of adenine to hypoxanthine by a manganese-dependent adenine deaminase encoded by a cryptic gene, yicP, which we propose be renamed ade. Insertion elements, located from -145 to +13 bp relative to the transcription start site, activated the ade gene as did unlinked mutations in the hns gene, encoding the histone-like protein H-NS. Gene fusion analysis indicated that ade transcription is repressed more than 10-fold by H-NS and that a region of 231 bp including the ade promoter is sufficient for this regulation. The activating insertion elements essentially eliminated the H-NS-mediated silencing, and stimulated ade gene expression 2-3-fold independently of the H-NS protein. PMID- 12077138 TI - FEEL-1, a novel scavenger receptor with in vitro bacteria-binding and angiogenesis-modulating activities. AB - Employing the expression cloning technique, we cloned a novel scavenger receptor that is structurally unrelated to other scavenger receptors. The cloned receptor contained fasciclin (Fas-1), epidermal growth factor (EGF)-like, laminin-type EGF like, and link domains. Based on the domain structures, we temporarily named it FEEL-1 (fasciclin, EGF-like, laminin-type EGF-like, and link domain-containing scavenger receptor-1). A data base search suggested the presence of a paralogous gene of FEEL-1, the full-length cDNA of this gene was also cloned, and its nucleotide sequence was determined. The deduced amino acid sequence of the clone indicated that its domain organization is similar to FEEL-1, and we named this clone FEEL-2. The effect of monoclonal antibodies against FEEL-1 indicated that FEEL-1 is the major receptor for 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbo cyanine perchlorate (DiI)-labeled acetylated low density lipoprotein (DiI-Ac-LDL) in human umbilical vein endothelial cells. Reverse transcription and PCR analysis revealed that both FEEL-1 and FEEL-2 were expressed in several tissues and expressed highly in the spleen and lymph node. On the other hand, only FEEL-1 was expressed in mononuclear cells, particularly resting CD14(+) cells. The transient expression of FEEL-1 and FEEL-2 in Chinese hamster ovary cells demonstrated that both FEELs could bind to DiI-Ac-LDL. Both receptors were also found to bind to Gram-negative and Gram-positive bacteria. These results suggest that FEELs play important roles in the defense mechanisms against bacterial infection. Finally, the phenotypic effect of the inhibition of FEEL-1 on vascular remodeling was tested in vitro using the Matrigel tube formation assay, and we found a marked reduction in the degree of cell-cell interaction in anti-FEEL-1 monoclonal antibody-treated cells, suggesting the role of this receptor in angiogenesis. PMID- 12077139 TI - Balance between two transpeptidation mechanisms determines the expression of beta lactam resistance in Enterococcus faecium. AB - The d,d-transpeptidase activity of high molecular weight penicillin-binding proteins (PBPs) is essential to maintain cell wall integrity as it catalyzes the final cross-linking step of bacterial peptidoglycan synthesis. We investigated a novel beta-lactam resistance mechanism involving by-pass of the essential PBPs by l,d-transpeptidation in Enterococcus faecium. Determination of the peptidoglycan structure by reverse phase high performance liquid chromatography coupled to mass spectrometry revealed that stepwise selection for ampicillin resistance led to the gradual replacement of the usual cross-links generated by the PBPs (d-Ala(4) -> d-Asx-Lys(3)) by cross-links resulting from l,d-transpeptidation (l-Lys(3) --> d-Asx-Lys(3)). This was associated with no modification of the level of production of the PBPs or of their affinity for beta-lactams, indicating that altered PBP activity was not required for ampicillin resistance. A beta-lactam insensitive l,d-transpeptidase was detected in membrane preparations of the parental susceptible strain. Acquisition of resistance was not because of variation of this activity. Instead, selection led to production of a beta-lactam insensitive d,d-carboxypeptidase that cleaved the C-terminal d-Ala residue of pentapeptide stems in vitro and caused massive accumulation of cytoplasmic precursors containing a tetrapeptide stem in vivo. The parallel dramatic increase in the proportion of l-Lys(3) --> d-Asx-Lys(3) cross-links showed that the enzyme was activating the resistance pathway by generating the substrate for the l,d transpeptidase. PMID- 12077140 TI - Beta-catenin N- and C-terminal tails modulate the coordinated binding of adherens junction proteins to beta-catenin. AB - beta-Catenin plays a central role in the establishment and regulation of adherens junctions because it interacts with E-cadherin and, through alpha-catenin, with the actin cytoskeleton. beta-Catenin is composed of three domains: a central armadillo repeat domain and two N- and C-terminal tails. The C-tail interacts with the armadillo domain and limits its ability to bind E-cadherin and other cofactors. The two beta-catenin tails are mutually inter-regulated because the C tail is also necessary for binding of the N-tail to the armadillo domain. Moreover, the N-tail restricts the interaction of the C-tail with the central domain. Depletion of either of the two tails has consequences for the binding of factors at the other end: deletion of the C-tail increases alpha-catenin binding, whereas deletion of the N-tail blocks E-cadherin interaction to the armadillo repeats. As an effect of the interconnection of the tails, the association of alpha-catenin and E-cadherin to beta-catenin is interdependent. Thus, binding of alpha-catenin to the N-tail, through conformational changes that affect the C tail, facilitates the association of E-cadherin. These results indicate that different cofactors of beta-catenin bind coordinately to this protein and indicate how the two terminal ends of beta-catenin exquisitely modulate intermolecular binding within junctional complexes. PMID- 12077141 TI - Structure-function relationships of the RNA-dependent RNA polymerase from poliovirus (3Dpol). A surface of the primary oligomerization domain functions in capsid precursor processing and VPg uridylylation. AB - The primary oligomerization domain of poliovirus polymerase, 3Dpol, is stabilized by the interaction of the back of the thumb subdomain of one molecule with the back of the palm subdomain of a second molecule, thus permitting the head-to-tail assembly of 3Dpol monomers into long fibers. The interaction of Arg-455 and Arg 456 of the thumb with Asp-339, Ser-341, and Asp-349 of the palm is key to the stability of this interface. We show that mutations predicted to completely disrupt this interface do not produce equivalent growth phenotypes. Virus encoding a polymerase with changes of both residues of the thumb to alanine is not viable; however, virus encoding a polymerase with changes of all three residues of the palm to alanine is viable. Biochemical analysis of 3Dpol derivatives containing the thumb or palm substitutions revealed that these derivatives are both incapable of forming long fibers, suggesting that polymerase fibers are not essential for virus viability. The RNA binding activity, polymerase activity, and thermal stability of these derivatives were equivalent to that of the wild-type enzyme. The two significant differences observed for the thumb mutant were a modest reduction in the ability of the altered 3CD proteinase to process the VP0/VP3 capsid precursor and a substantial reduction in the ability of the altered 3Dpol to catalyze oriI-templated uridylylation of VPg. The defect to uridylylation was a result of the inability of 3CD to stimulate this reaction. Because 3C alone can substitute for 3CD in this reaction, we conclude that the lethal replication phenotype associated with the thumb mutant is caused, in part, by the disruption of an interaction between the back of the thumb of 3Dpol and some undefined domain of 3C. We speculate that this interaction may also be critical for assembly of other complexes required for poliovirus genome replication. PMID- 12077142 TI - Functional conservation for lipid storage droplet association among Perilipin, ADRP, and TIP47 (PAT)-related proteins in mammals, Drosophila, and Dictyostelium. AB - Intracellular neutral lipid storage droplets are essential organelles of eukaryotic cells, yet little is known about the proteins at their surfaces or about the amino acid sequences that target proteins to these storage droplets. The mammalian proteins Perilipin, ADRP, and TIP47 share extensive amino acid sequence similarity, suggesting a common function. However, while Perilipin and ADRP localize exclusively to neutral lipid storage droplets, an association of TIP47 with intracellular lipid droplets has been controversial. We now show that GFP-tagged TIP47 co-localizes with isolated intracellular lipid droplets. We have also detected a close juxtaposition of TIP47 with the surfaces of lipid storage droplets using antibodies that specifically recognize TIP47, further indicating that TIP47 associates with intracellular lipid storage droplets. Finally, we show that related proteins from species as diverse as Drosophila and Dictyostelium can also target mammalian or Drosophila lipid droplet surfaces in vivo. Thus, sequence and/or structural elements within this evolutionarily ancient protein family are necessary and sufficient to direct association to heterologous intracellular lipid droplet surfaces, strongly indicating that they have a common function for lipid deposition and/or mobilization. PMID- 12077143 TI - Effects of hydrogen bonding within a damaged base pair on the activity of wild type and DNA-intercalating mutants of human alkyladenine DNA glycosylase. AB - Human alkyladenine DNA glycosylase "flips" damaged DNA bases into its active site where excision occurs. Tyrosine 162 is inserted into the DNA helix in place of the damaged base and may assist in nucleotide flipping by "pushing" it. Mutating this DNA-intercalating Tyr to Ser reduces the DNA binding and base excision activities of alkyladenine DNA glycosylase to undetectable levels demonstrating that Tyr-162 is critical for both activities. Mutation of Tyr-162 to Phe reduces the single turnover excision rate of hypoxanthine by a factor of 4 when paired with thymine. Interestingly, when the base pairing partner for hypoxanthine is changed to difluorotoluene, which cannot hydrogen bond to hypoxanthine, single turnover excision rates increase by a factor of 2 for the wild type enzyme and about 3 to 4 for the Phe mutant. In assays with DNA substrates containing 1,N(6) ethenoadenine, which does not form hydrogen bonds with either thymine or difluorotoluene, base excision rates for both the wild type and Phe mutant were unaffected. These results are consistent with a role for Tyr-162 in pushing the damaged base to assist in nucleotide flipping and indicate that a nucleotide flipping step may be rate-limiting for excision of hypoxanthine. PMID- 12077144 TI - Regulation of the human cyclin-dependent kinase inhibitor p18INK4c by the transcription factors E2F1 and Sp1. AB - The p18(INK4c) cyclin-dependent kinase inhibitor is an important regulator of cell cycle progression and cellular differentiation. We and others found that overexpressed E2F proteins up-regulate p18 expression. To better understand this phenomenon, we performed a functional analysis of the human p18 promoter. Deletion studies revealed that the E2F-responsive elements of the promoter are located within 131 bp upstream of the transcription start site. This region contains putative Sp1- and E2F-binding sites. Mutational inactivation of these elements revealed that the Sp1 sites were important for the basal activity of the promoter but could also mediate the effects of E2F1 on the p18 promoter. Moreover, we found that E2F1 and Sp1 can synergistically enhance the activity of the proximal p18 promoter. Gel shift analyses using p18 promoter-derived probes led to the identification of several multiprotein complexes that were found to contain different combinations of E2F proteins and/or Sp1. Recombinant E2F1 was also capable of binding to the E2F-binding sites. Chromatin immunoprecipitation experiments demonstrated that E2F1 and E2F4 associate with the p18 promoter in unperturbed cells. Based on these findings, we conclude that E2F proteins and Sp1 play an important role in the control of p18 expression. PMID- 12077145 TI - Transcriptional profiling identifies two members of the ATP-binding cassette transporter superfamily required for sterol uptake in yeast. AB - In contrast to lipoprotein-mediated sterol uptake, free sterol influx by eukaryotic cells is poorly understood. To identify components of non-lipoprotein mediated sterol uptake, we utilized strains of Saccharomyces cerevisiae that accumulate exogenous sterol due to a neomorphic mutation in the transcription factor, UPC2. Two congenic upc2-1 strains, differing quantitatively in aerobic sterol uptake due to a modifying mutation in the HAP1 transcription factor, were compared using DNA microarrays. We identified 9 genes as responsive to UPC2 that were also induced under anaerobiosis, when sterol uptake is essential. Deletion mutants in these genes were assessed for sterol influx in the upc2-1 background. UPC2 itself was up-regulated under these conditions and was required for aerobic sterol influx. Deletion of the ATP-binding cassette transporters YOR011w (AUS1) or PDR11, or a putative cell wall protein encoded by DAN1, significantly reduced sterol influx. Sodium azide and vanadate inhibited sterol uptake, consistent with the participation of ATP-binding cassette transporters. We hypothesized that the physiological role of Aus1p and Pdr11p is to mediate sterol uptake when sterol biosynthesis is compromised. Accordingly, expression of AUS1 or PDR11 was required for anaerobic growth and sterol uptake. We proposed similar molecules may be important components of sterol uptake in all eukaryotes. PMID- 12077146 TI - Identification of a signal transduction switch in the chemokine receptor CXCR1. AB - Chemokine receptors belong to the superfamily of G protein-coupled receptors, which regulate the trafficking and activation of leukocytes, and operate as coreceptors in the entry of HIV-1. To investigate the early steps in the signal transmission from the chemokine-binding site to the G protein-coupling region we engineered metal ion-binding sites at putative extracellular sites in the chemokine receptor CXCR1. We introduced histidines into sites located in the second and third putative extracellular loops of CXCR1, creating single, double, and triple mutant receptors: R199H, R203H, D265H, R199H/R203H, R199H/D265H, R203H/D265H, R203H/H207Q, and R199H/R203H/D265H. Cells expressing the double mutants R199H/D265H and R203H/D265H and the triple mutant R199H/R203H/D265H failed to trigger interleukin 8-dependent calcium responses. Interestingly, calcium responses mediated by the single mutant R203H and the double mutants R199H/R203H and R203H/H207Q were blocked by Zn(II), indicating the creation of a functional metal ion-binding site. On the other hand, cells expressing all single, double, or triple histidine-substituted CXCR1 demonstrated high affinity binding to interleukin 8 in the presence and absence of metal ions. These findings indicate that occupation of the engineered metal-binding site uncouples the chemokine-binding site from the activation mechanism in CXCR1. Most importantly, we identify for the first time elements of an early signal transduction switch of chemokine receptors before the activation of cytoplasmic G proteins. PMID- 12077147 TI - Induction of MUC2 and MUC5AC mucins by factors of the epidermal growth factor (EGF) family is mediated by EGF receptor/Ras/Raf/extracellular signal-regulated kinase cascade and Sp1. AB - The 11p15 mucin genes (MUC2, MUC5AC, MUC5B and MUC6) possess a cell-specific pattern of expression in normal lung that is altered during carcinogenesis. Growth factors of the epidermal growth factor family are known to target key genes that in turn may affect the homeostasis of lung mucosae. Our aim was to study the regulation of the 11p15 mucin genes both at the promoter and protein levels to assess whether their altered expression may represent a key event during lung carcinogenesis. Studies were performed in the mucoepidermoid NCI-H292 lung cancer cell line. Cell treatment with epidermal growth factor (EGF), transforming growth factor alpha (TGF-alpha), or tumor necrosis factor alpha (TNF alpha) resulted in a dramatic increase of MUC2 and MUC5AC mRNAs levels, promoter activity, and apomucin expression, whereas those of MUC5B and MUC6 were unchanged. pGL3 deletion mutants of MUC2, MUC5AC, and MUC5B promoters were constructed and used in transient transfection assays to characterize EGF- and TGF-alpha-responsive regulatory regions within the promoters. They were located in the -2627/-2097 and -202/-1 regions of MUC2 and MUC5AC promoters, respectively. Finally, we demonstrate that transcription factor Sp1 not only binds and activates MUC2 and MUC5AC promoters but also participates to their EGF- and TGF-alpha-mediated up-regulation. We also show that Sp3 is a strong inhibitor of 11p15 mucin gene transcription. In conclusion, MUC2 and MUC5AC are two target genes of EGFR ligands in lung cancer cells, and up-regulation of these two genes goes through concomitant activation of the EGFR/Ras/Raf/Extracellular Signal regulated Kinase-signaling pathway and Sp1 binding to their promoters. PMID- 12077148 TI - Heparin/Heparan sulfate domains in binding and signaling of fibroblast growth factor 8b. AB - The role of heparin and heparan sulfate in the binding and signaling of fibroblast growth factors (FGFs) has been subject to intense investigation, but the studies have largely been confined to two species (FGF1 and FGF2) of the family with approximately 20 members. We have investigated the structural requirements for heparin/heparan sulfate in binding and activation of FGF8 (splice variant b). We present evidence that the minimal FGF8b-binding saccharide domain encompasses 5-7 monosaccharide units. The N-, 2-O-, and 6-O-sulfate substituents of heparin/heparan sulfate (HS) are all involved in the interaction, preferentially in the form of trisulfated IdoUA(2-OSO(3))-GlcNSO(3)(6-OSO(3)) disaccharide constituents. These structural characteristics resemble those described earlier for FGF1. By contrast, the saccharide structures required for the biological activity of FGF8b differed significantly from those characteristic for FGF1 and FGF2. Experiments with cells lacking active HS indicated that extended >/=14-mer heparin domains were needed to enhance cell proliferation and Erk phosphorylation by FGF8b, whereas in cells stimulated with FGF1 or FGF2 the corresponding responses were achieved by much shorter, 6-8-mer, oligosaccharides. Furthermore, still longer domains were needed to activate FGF8b in cells with "non-optimal" FGF receptor expression. Collectively, our data suggest that the heparin/HS structures enhancing the biological activity of FGFs were influenced by the FGF species involved as well as by the cellular composition of FGF receptors. PMID- 12077149 TI - (-)-Doliculide, a new macrocyclic depsipeptide enhancer of actin assembly. AB - The cytotoxic, cyclic depsipeptide (-)-doliculide was originally isolated by Ishiwata et al. (Ishiwata, H., Nemoto, T., Ojika, M., and Yamada, K. (1994) J. Org. Chem. 59, 4710-4711 and Ishiwata, H., Sone, H., Kigoshi, H., and Yamada, K. (1994) J. Org. Chem. 59, 4712-4713) from the sea hare Dolabella auricularia collected in Japanese waters, but the mechanism of action of the depsipeptide was not known. Using synthetic (-)-doliculide, we found that the compound arrests cells at the G(2)/M phase of the cell cycle by interfering with normal actin assembly. In cells, normal stress fibers disappeared and were replaced by multiple clumps of apparently aggregated F-actin. These effects of (-)-doliculide on cells were essentially identical to those obtained with jasplakinolide. Like jasplakinolide, (-)-doliculide caused the hyperassembly of purified actin into F actin as measured both fluorometrically and by centrifugation. In addition, (-) doliculide, like jasplakinolide, readily displaced a fluorescent phalloidin derivative from actin polymer. In these biochemical assays (-)-doliculide and jasplakinolide were quantitatively virtually identical in their behaviors. Similar effects have also been reported with a series of depsipeptides known as chondramides. Using recently developed, computer-driven shape descriptor analysis (Mansfield, M. L., Covell, D. G., and Jernigan, R. L. (2002) J. Chem. Inf. Comput. Sci. 42, 259-273), we compared (-)-doliculide with jasplakinolide, phalloidin, and chondramide C to gain insight into a possible pharmacophore that would explain the apparent binding of this diverse group of molecules at the same site on F-actin. We found that the segment of (-)-doliculide that best overlapped the other molecules encompassed its phenyl and isopropyl side chains and the portion of the macrocycle between these substituents. PMID- 12077150 TI - Reconstitution of enzymatic activity by the association of the cap and catalytic domains of human topoisomerase I. AB - When human topoisomerase I binds DNA, two opposing lobes in the enzyme, the cap region (amino acid, residues 175-433) and the catalytic domain (Deltacap, residues 433 to the COOH terminus) clamp tightly around the DNA helix to form the precleavage complex. Although Deltacap contains all of the residues known to be important for catalysis and binds DNA with an affinity similar to that of the intact enzyme, this fragment lacks catalytic activity. However, a mixture of Deltacap and topo31 (residues 175-433) reconstitutes enzymatic activity as measured by plasmid DNA relaxation and suicide cleavage assays. Although the formation of an active complex between topo31 and Deltacap is too unstable to be detected by pull-down experiments even in the presence of DNA, the association of topo31 with Deltacap persists and is detectable after the complex catalyzes the covalent attachment of the DNA to Deltacap by suicide cleavage. Removal of topo31 from Deltacap-DNA after suicide cleavage reveals that, unlike the cleavage reaction, religation does not require the cap region of the protein. These results suggest that activation of the catalytic domain of the enzyme for cleavage requires both DNA binding and the presence of the cap region of the protein. PMID- 12077151 TI - Profiling membrane lipids in plant stress responses. Role of phospholipase D alpha in freezing-induced lipid changes in Arabidopsis. AB - A sensitive approach based on electrospray ionization tandem mass spectrometry has been employed to profile membrane lipid molecular species in Arabidopsis undergoing cold and freezing stresses. Freezing at a sublethal temperature induced a decline in many molecular species of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylglycerol (PG) but induced an increase in phosphatidic acid (PA) and lysophospholipids. To probe the metabolic steps generating these changes, lipids of Arabidopsis deficient in the most abundant phospholipase D, PLD alpha, were analyzed. The PC content dropped only half as much, and PA levels rose only half as high in the PLD alpha-deficient plants as in wild-type plants. In contrast, neither PE nor PG levels decreased significantly more in wild-type plants than in PLD alpha-deficient plants. These data suggest that PC, rather than PE and PG, is the major in vivo substrate of PLD alpha. The action of PLD alpha during freezing is of special interest because Arabidopsis plants that are deficient in PLD alpha have improved tolerance to freezing. The greater loss of PC and increase in PA in wild-type plants as compared with PLD alpha-deficient plants may be responsible for destabilizing membrane bilayer structure, resulting in a greater propensity toward membrane fusion and cell death in wild-type plants. PMID- 12077152 TI - The roadblock light chain binds a novel region of the cytoplasmic Dynein intermediate chain. AB - Cytoplasmic dynein is the major minus-end directed microtubule-based motor in eukaryotic cells. It is composed of a number of different subunits including three light chain families: Tctex1, LC8, and roadblock. The incorporation of the roadblock light chains into the cytoplasmic dynein complex had not been determined. There are two roadblock genes in mammals, ROBL-1 and ROBL-2. We find that both members of the roadblock family bind directly to all of the intermediate chain isoforms of mammalian cytoplasmic dynein. This was determined with three complementary approaches. A yeast two-hybrid assay demonstrated that both roadblock light chains interact with intermediate chain isoforms from the IC74-1 and IC74-2 genes in vivo. This was confirmed in vitro with both a solid phase blot overlay assay and a solution-binding assay. The roadblock-binding domain on the intermediate chain was mapped to an approximately 72 residue region. The binding domain is downstream of each of the two alternative splice sites in the intermediate chains. This location is consistent with the finding that both roadblock-1 and roadblock-2 show no binding specificity for a single IC74-1 or IC74-2 intermediate chain isoform. In addition, this roadblock-binding domain is significantly downstream from both the Tctex1- and LC8-binding sites, supporting the hypothesis that multiple light chain family members can bind to the same intermediate chain. PMID- 12077153 TI - Global antibiotic resistance in Streptococcus pneumoniae. AB - The last two decades of the 20th century were marked by an increasing resistance rate among several bacteria. Threat of resistance is present in Staphylococcus spp., Enterococcus spp., Pseudomonas spp. and Enterobacteriaceae, which are the major pathogens in nosocomial infections. In the community, too, increasing resistance can be observed and is attributed mainly (but not exclusively) to Streptococcus pneumoniae and Haemophilus influenzae. To scrutinize this trend, resistance surveillance in the community was established about 10 years ago. One of the multinational, longitudinal surveillance programmes in place is the Alexander Project, which was established in 1992 to monitor the susceptibility of the major community-acquired lower respiratory tract pathogens to a range of antibacterial drugs. The Alexander Project has revealed a tendency towards increasing resistance of S. pneumoniae to penicillin and macrolide therapy. Within Europe, the prevalence of penicillin resistance among S. pneumoniae isolates is particularly high in France and Spain. Macrolide resistance in S. pneumoniae is also a growing problem in European countries such as France, Spain, Belgium and Italy, where the extent of macrolide resistance in S. pneumoniae now exceeds that of penicillin resistance. PMID- 12077154 TI - Antibacterial activity of oral antibiotics against community-acquired respiratory pathogens from three European countries. AB - Antimicrobial resistance is universally recognized as a major problem. A European resistance survey was established to monitor the activity of widely used oral antibiotics against common respiratory tract pathogens. Studies were conducted in Italy, Spain and Austria to monitor resistance patterns among respiratory Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pyogenes, Staphylococcus aureus and Klebsiella pneumoniae to amoxicillin, co-amoxiclav, penicillin, cefaclor, cefadroxil, cefalexin, cefprozil, cefuroxime, cefixime, ceftibuten, cefpodoxime, clarithromycin and azithromycin (the antibiotics tested varying slightly from country to country). Minimum inhibitory concentrations were determined using the NCCLS-recommended broth microdilution method. Among the antibiotics tested, cefpodoxime, an oral cephalosporin, was remarkably active against the major respiratory pathogens in all three countries. Cefpodoxime was more potent than cefaclor, cefixime and ceftibuten against pneumococci, especially against strains with decreased sensitivity to penicillin, and more active than cefaclor and cefuroxime against Gram-negative respiratory pathogens. Pneumococci and staphylococci displayed a very high level of in vitro macrolide resistance. These data indicate that cefpodoxime represents an appropriate choice in the treatment of community acquired respiratory tract infection in the three countries surveyed. PMID- 12077155 TI - Pharmacokinetics and pharmacodynamics of oral beta-lactam antibiotics as a two dimensional approach to their efficacy. AB - Pharmacokinetic and pharmacodynamic parameters are increasingly recognized as important determinants of the therapeutic efficacy of an antibiotic. For beta lactam antibiotics, the most important determinant of the antimicrobial efficacy, and hence predictor of therapeutic efficacy, is the length of time that serum concentrations exceed the MIC. Dosing schedules for beta-lactam antibiotics should maintain serum concentrations above the MIC for the bacterial pathogen for at least 50% of the dosing interval to achieve therapeutic efficacy and prevent the development of resistance. This is a basic criterion for the clinical efficacy of beta-lactams. A combination of microbiological activity and pharmacokinetic characteristics was applied to calculate the time that serum antibiotic concentrations exceed the MIC for the major respiratory tract pathogens such as Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pyogenes and Klebsiella pneumoniae. In contrast with some other oral beta-lactam antibiotics, cefpodoxime 200 mg bd maintains serum concentrations above the MIC for each organism for at least 50% of the dosing interval and may therefore be an attractive choice for empirical therapy of community-acquired lower respiratory tract infections. PMID- 12077156 TI - Interstitial tissue concentrations of cefpodoxime. AB - Microdialysis is a technique that allows the measurement of concentrations of free antibiotic in tissue. The free antibiotic concentration is responsible for the antibacterial effect at the target site. We used microdialysis in animal and human studies to investigate the tissue penetration of cefpodoxime. In the animal study, total plasma and free muscle and lung concentrations of cefpodoxime were measured after male Wistar rats had received either 10 mg/kg or 20 mg/kg i.v. cefpodoxime over 5 h or a continuous i.v. infusion of 260 microg/h cefpodoxime after a loading dose of 6 mg/kg. Free muscle concentrations of cefpodoxime were similar to free lung concentrations and therefore provided a surrogate measure of cefpodoxime concentrations at the pulmonary target site. In an open, randomized, two-way crossover, single-dose study in six healthy male volunteers, total plasma and free muscle concentrations were measured after a single oral dose of cefpodoxime 400 mg or cefixime 400 mg. The total plasma concentrations of each antibiotic were similar and higher than free muscle concentrations. The tissue penetration of cefpodoxime was, however, greater than that of cefixime, as shown by two-fold higher peak free muscle concentrations after dosing with cefpodoxime than with cefixime (2.1 mg/L versus 0.9 mg/L). In addition, the area under the curve for tissue (AUC(t)) of cefpodoxime (400 mg) was more than double that of cefixime (400 mg), based on free antibiotic concentrations (15.4 mg x h/L versus 7.3 mg x h/L). These findings indicate that, taking into account pharmacokinetic/pharmacodynamic considerations, cefpodoxime is likely to be more efficacious than cefixime, due to its greater tissue penetration. PMID- 12077157 TI - Clinical efficacy of cefpodoxime in respiratory tract infection. AB - Acute otitis media (AOM), sinusitis and tonsillopharyngitis are respiratory tract infections frequently encountered by primary-care physicians. Increasing bacterial resistance, particularly in Streptococcus pneumoniae, which is one of the most important respiratory tract bacteria implicated in community-acquired respiratory tract infections, has led to concern about the current options for empirical antibiotic treatment and has prompted a search for effective alternative treatments. Data from in vitro studies show that cefpodoxime has good activity against the main respiratory tract pathogens, S. pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pyogenes. Clinical studies confirm the efficacy of cefpodoxime in AOM, sinusitis and tonsillopharyngitis. As with all broad-spectrum antibiotics, there is the risk of promotion of bacterial resistance associated with overuse. However, if used with care, cefpodoxime can be considered as an alternative for empirical treatment of bacterial respiratory tract infections encountered in general practice, particularly where penicillins and macrolides have reduced efficacy against the main bacterial pathogens. PMID- 12077159 TI - Temperature-dependence of mitochondrial function and production of reactive oxygen species in the intertidal mud clam Mya arenaria. AB - Mitochondrial respiration, energetic coupling to phosphorylation and the production of reactive oxygen species (ROS) were studied in mitochondria isolated from the eurythermal bivalve Mya arenaria (Myoidea) from a low-shore intertidal population of the German Wadden Sea. Measurements were conducted both within the range of the habitat temperatures (5-15 degrees C) and when subjected to heat exposure at 20 degrees C and 25 degrees C. Experimental warming resulted in an increase in the rate of state 3 and state 4 respiration in isolated mitochondria. The highest respiratory coupling ratios (RCR) were found at 15 degrees C; at higher temperatures mitochondrial coupling decreased, and release of ROS doubled between 15 and 25 degrees C. ROS production was 2-3% of total oxygen consumption in state 3 (0.3-0.5 nmol ROS mg(-1) protein min(-1)) at the habitat temperature, reaching a maximum of 4.3 % of state 3 respiration and 7 % of oligomycin-induced state 4+ respiration under heat stress. Thus, state 4 respiration, previously interpreted exclusively as a measure of proton leakage, included a significant contribution from ROS formation in this animal, especially under conditions of heat stress. Oxygen radical formation was directly dependent on temperature controlled respiration rates in states 3 and 4 and inversely related to mitochondrial coupling (RCR+) in state 4. Mitochondrial ROS formation is therefore involved in cellular heat stress in this eurythermal marine ectotherm. PMID- 12077160 TI - Delayed depolarization of the cog-wheel valve and pulmonary-to-systemic shunting in alligators. AB - Alligators and other crocodilians have a cog-wheel valve located within the subpulmonary conus, and active closure of this valve during each heart beat can markedly and phasically increase resistance in the pulmonary outflow tract. If this increased resistance causes right ventricular pressure to rise above that in the systemic circuit, right ventricular blood can flow into the left aorta and systemic circulation, an event known as pulmonary-to-systemic shunting. To understand better how this valve is controlled, anaesthetized American alligators (Alligator mississippiensis) were used to examine the relationships between depolarization of the right ventricle, depolarization/contraction of the cog wheel valve muscle and the resultant right ventricular, pulmonary artery and systemic pressures. Depolarization swept across the right ventricle from the apex towards the base (near where the cog-wheel valve muscle is located) at a velocity of 91+/-23 cm s(-1) (mean +/- S.E.M., N=3). The cog-wheel valve electrocardiogram (ECG) (and thus contraction of the valve) trailed the right ventricular ECG by 248+/-28 ms (N=3), which was equivalent to 6-35 % of a cardiac cycle. This long interval between right ventricular and valve depolarization suggests a nodal delay at the junction between the base of the right ventricle and the cog-wheel valve. The delay before valve closure determined when the abrupt secondary rise in right ventricular pressure occurred during systole and is likely to strongly influence the amount of blood entering the pulmonary artery and thus to directly control the degree of shunting. Left vagal stimulation (10-50 Hz) reduced the conduction delay between the right ventricle and cog-wheel valve by approximately 20 % and reduced the integrated cog-wheel ECG by 10-20 %. Direct application of acetylcholine (1-2 mg) also reduced the integrated cog-wheel ECG by 10-100 %; however, its effect on the conduction delay was highly variable (-40 to +60 %). When the cog-wheel valve muscle was killed by the application of ethanol, the cog wheel ECG was absent, right ventricular and pulmonary pressures remained low and tracked one another, the secondary rise in right ventricular pressure was abolished and shunting did not occur. This study provides additional, direct evidence that phasic contraction of the cog-wheel valve muscle controls shunting, that nervous and cholinergic stimulation can alter the delay and strength of valve depolarization and that this can affect the propensity to shunt. PMID- 12077161 TI - Salt and water regulation by the leatherback sea turtle Dermochelys coriacea. AB - We measured the salt and water balance of hatchling leatherback sea turtles, Dermochelys coriacea, during their first few days of life to investigate how they maintain homeostasis under the osmoregulatory challenge of a highly desiccating terrestrial environment and then a hyperosmotic marine environment. Hatchlings desiccated rapidly when denied access to sea water, with their hematocrit increasing significantly from 30.32+/-0.54 % to 38.51+/-1.35 % and plasma Na(+) concentration increasing significantly from 138.2+/-3.3 to 166.2+/-11.2 mmol l( 1) in 12 h. When hatchlings were subsequently put into sea water, hematocrit decreased and plasma Na(+) concentration was unchanged but both were significantly elevated above pretreatment values. In other hatchlings kept in sea water for 48 h, body mass and plasma Na(+) concentration increased significantly, but hematocrit did not increase. These data show that hatchlings were able to osmoregulate effectively and gain mass by drinking sea water. We stimulated hatchlings to secrete salt from the salt glands by injecting a salt load of 27 mmol kg(-1). The time taken for secretion to begin in newly hatched turtles was longer than that in 4-day-old hatchlings, but the secretory response was identical at 4.15+/-0.40 and 4.13+/-0.59 mmol Na(+) kg(-1) h(-1) respectively. Adrenaline and methacholine were both potent inhibitors of salt gland secretion in a dose-dependent manner, although methacholine administered simultaneously with a subthreshold salt load elicited a transient secretory response. The results showed that hatchling leatherbacks are able to tolerate significant changes in internal composition and efficiently use their salt glands to establish internal ionic and water balance when in sea water. PMID- 12077162 TI - Influence of cues from the anterior medial eyes of virtual prey on Portia fimbriata, an araneophagic jumping spider. AB - Portia fimbriata from Queensland, Australia, is a jumping spider (Salticidae) that preys on other spiders, including other salticids. Cryptic stalking (palps retracted, walking very slowly and freezing when faced) is a prey-specific tactic deployed exclusively against salticid prey. Using vision alone, P. fimbriata discriminates salticid from non-salticid prey, with the prey salticid's large anterior median (AM) eyes providing critical cues. Here, using computer-rendered virtual three-dimensional lures, we clarify experimentally some of the specific optical cues from AM eyes that influence the behaviour of P. fimbriata. Control lures were based on Jacksonoides queenslandicus, the salticid upon which P. fimbriata most commonly preys in nature. Experimental lures were modified to isolate specific combinations of AM eye features. For presentation to P. fimbriata, lures were projected on a small screen positioned in front of a web covered platform. Each individual P. fimbriata was tested once with a control lure and once with a lure on which some AM eye feature had been altered. P. fimbriata was affected by the presence-versus-absence of AM eyes, by the size and shape of these eyes and by AM eye position. Horizontal position on the face of a single AM eye had no discernible influence on whether P. fimbriata initiated cryptic stalking, but reduced how often P. fimbriata froze when faced by the prey. The implications of the findings are discussed in relation to perceptual processes underlying the predatory strategy of P. fimbriata. PMID- 12077163 TI - Neurochemical fine tuning of a peripheral tissue: peptidergic and aminergic regulation of fluid secretion by Malpighian tubules in the tobacco hawkmoth M. sexta. AB - The actions of various peptides and other compounds on fluid secretion by Malpighian tubules in the tobacco hawkmoth Manduca sexta sexta are investigated in this study. Using a newly developed pharate adult Malpighian tubule bioassay, we show that three tachykinin-related peptides (TRPs), leucokinin I, serotonin (5 HT), octopamine, the cardioacceleratory peptides 1a, 1b and 2c, cGMP and cAMP each cause an increase in the rate of fluid secretion in pharate adult tubules. Whereas the possible hormonal sources of biogenic amines and some of the peptides are known, the distribution of TRPs has not been investigated previously in M. sexta. Thus we performed immunocytochemistry using an anti-TRP antiserum. We show the presence of TRP-like material in a small subset of cells in the M. sexta central nervous system (CNS). The larval brain contains approximately 60 TRP immunopositive cells and there are approximately 100 such cells in the adult brain including the optic lobes. Every ganglion of the ventral nerve cord also contains TRP-like immunoreactive cells. No TRP-containing neurosecretory cells were seen in the CNS, but endocrine cells of the midgut reacted with the antiserum. We propose the hypothesis that the control in insects of physiological systems by hormones may not always involve tissue-specific hormones that force stereotypical responses in their target systems. Instead, there may exist in the extracellular fluid a continuous broadcast of information in the form of a chemical language to which some or all parts of the body continuously respond on a moment-to-moment basis, and which ensures a more effective and efficient coordination of function than could be achieved otherwise. PMID- 12077164 TI - Effects of temperature, epinephrine and Ca(2+) on the hearts of yellowfin tuna (Thunnus albacares). AB - Tuna are endothermic fish with high metabolic rates, cardiac outputs and aerobic capacities. While tuna warm their skeletal muscle, viscera, brain and eyes, their hearts remain near ambient temperature, raising the possibility that cardiac performance may limit their thermal niches. We used an in situ perfused heart preparation to investigate the effects of acute temperature change and the effects of epinephrine and extracellular Ca(2+) on cardiac function in yellowfin tuna (Thunnus albacares). Heart rate showed a strong temperature-dependence, ranging from 20 beats min(-1) at 10 degrees C to 109 beats min(-1) at 25 degrees C. Maximal stroke volume showed an inverse temperature-dependence, ranging from 1.4 ml kg(-1) at 15 degrees C to 0.9 ml kg(-1) at 25 degrees C. Maximal cardiac outputs were 27 ml kg(-1) min(-1) at 10 degrees C and 98 ml kg(-1) min(-1) at 25 degrees C. There were no significant effects of perfusate epinephrine concentrations between 1 and 100 nmol l(-1) at 20 degrees C. Increasing extracellular Ca(2+) concentration from 1.84 to 7.36 mmol l(-1) at 20 degrees C produced significant increases in maximal stroke volume, cardiac output and myocardial power output. These data demonstrate that changes in heart rate and stroke volume are involved in maintaining cardiac output during temperature changes in tuna and support the hypothesis that cardiac performance may limit the thermal niches of yellowfin tuna. PMID- 12077165 TI - A Ca(2+)-sensing receptor modulates shark rectal gland function. AB - The elasmobranch Squalus acanthias controls plasma osmolality and extracellular fluid volume by secreting a hypertonic fluid from its rectal gland. Because we found a correlation between extracellular Ca(2+) concentration and changes in cytosolic Ca(2+) ([Ca(2+)](i)), we sought the possible presence of a calcium sensing receptor in rectal gland artery and tubules. Cytosolic Ca(2+) of both tissues responded to the addition of external Ca(2+) (0.8-5.3 mmol l(-1)) in a linear fashion. Spermine, Gd(3+) and Ni(2+), known agonists of the calcium sensing receptor, increased [Ca(2+)](i). To assess the participation of inositol triphosphate (IP(3)) generation, sarcoplasmic/endoplasmic reticulum (SR/ER) Ca(2+) depletion, and activation of store-operated Ca(2+) entry, we utilized thapsigargin and ryanodine to deplete Ca(2+) SR/ER stores and the inhibitory reagents TMB-8 and 2-APB to block IP(3) receptors. In each case, these agents inhibited the [Ca(2+)](i) response to agonist stimulation by approximately 50 %. Blockade of L-channels with nifedipine had no significant effect. Increases in ionic strength are known to inhibit the calcium-sensing receptor. We postulate that the CaSR stimulates Ca(2+)-mediated constriction of the rectal gland artery and diminishes cyclic AMP-mediated salt secretion in rectal gland tubules during non-feeding conditions. When the shark ingests sea water and fish, an increase in blood and interstitial fluid ionic strength inhibits the activity of the calcium sensing receptor, relaxing the rectal gland artery and permitting salt secretion by the rectal gland tubules. PMID- 12077166 TI - Sperm whale sound production studied with ultrasound time/depth-recording tags. AB - Delphinoids (Delphinidae, Odontoceti) produce tonal sounds and clicks by forcing pressurized air past phonic lips in the nasal complex. It has been proposed that homologous, hypertrophied nasal structures in the deep-diving sperm whale (Physeter macrocephalus) (Physeteridae, Odontoceti) are dedicated to the production of clicks. However, air volumes in diving mammals are reduced with increasing ambient pressure, which seems likely to influence pneumatic sound production at depth. To study sperm whale sound production at depth, we attached ultrasound time/depth-recording tags to sperm whales by means of a pole and suction cup. We demonstrate that sperm whale click production in terms of output and frequency content is unaffected by hydrostatic reduction in available air volume down to less than 2% of the initial air volume in the nasal complex. We present evidence suggesting that the sound-generating mechanism has a bimodal function, allowing for the production of clicks suited for biosonar and clicks more suited for communication. Shared click features suggest that sound production in sperm whales is based on the same fundamental biomechanics as in smaller odontocetes and that the nasal complexes are therefore not only anatomically but also functionally homologous in generating the initial sound pulse. PMID- 12077167 TI - Fast muscle in squid (Loligo pealei): contractile properties of a specialized muscle fibre type. AB - The contractile properties of the transverse muscle of the tentacles and the transverse muscle of the arms of the squid Loligo pealei were investigated using small muscle fibre bundle preparations. In addition, transmission electron microscopy was used to measure the length of the thick myofilaments of the two muscle fibre types. The thick filament length of the cross-striated tentacle fibres was 0.81+/-0.08 microm (mean +/- S.D, N=51) while that of the obliquely striated arm muscle fibres was 7.41+/-0.44 microm (N=58). The difference in thick filament length of the two muscle types was predicted to result in a much higher shortening velocity of the tentacle muscle compared with the arm muscle. This was tested by investigating the force/velocity relationship for isotonic shortening of the two muscle types. Fitting Hill's equation to the results gave a maximum shortening velocity (V(max), the intercept on the velocity axis) of 15.4+/-1.0 L(0) s(-1) (mean +/- S.D., N=9) for the tentacle fibres and of 1.5+/-0.2 L(0) s( 1) (N=8) for the arm fibres, where L(0) is the length at which peak isometric force was recorded. The difference in thick filament length was also predicted to result in lower peak tension in the tentacle versus the arm muscle. For the tentacle, the mean peak tetanic tension during a brief isometric tetanus (0.2s) of 131+/-56 mN mm(-2) cross-sectional area (mean +/- S.D., N=12) was observed at a stimulus frequency of 80 Hz, whereas the mean peak tetanic tension of the arm fibres during a brief isometric tetanus (0.2s) was 468+/-91 mN mm(-2) (N=5) and was observed at a stimulus frequency of 160 Hz. The length/force relationships (expressed relative to L(0)) of the two muscle types were similar. The ratio of twitch force to peak tetanic force was 0.66 in the tentacle fibres, but only 0.03 in the arm fibres. PMID- 12077168 TI - The relationship between heart rate and rate of oxygen consumption in Galapagos marine iguanas (Amblyrhynchus cristatus) at two different temperatures. AB - To enable the use of heart rate (fH) for estimating field metabolic rate (FMR) in free-ranging Galapagos marine iguanas Amblyrhynchus cristatus, we determined the relationships between fH and mass-specific rate of oxygen consumption (sVO2) in seven iguanas before and during exercise on a treadmill and during the post exercise period. The experiments were conducted at 27 and 35 degrees C, which are the temperatures that represent the lowest and highest average body temperatures of these animals in the field during summer. There were linear and significant relationships between fH and sVO2 at both temperatures (r(2)=0.86 and 0.91 at 27 degrees C and 36 degrees C, respectively). The slopes of the two regression lines did not differ, but there were significant differences in their intercepts. Thus, while heart rate can be used to predict FMR, the effects of temperature on the intercept of the regression must be taken into account when converting fH to sVO2. On the basis of our data, this can be achieved by applying the following formula: sVO2=0.0113fH-0.2983Q(10)((T(b)-27)/10). The increase in sVO2 with elevated body temperature results from an increase in fH, with no significant change in mass-specific oxygen pulse (sO(2) pulse; cardiac stroke volume times the difference in oxygen content between arterial and mixed venous blood). However, during exercise at both temperatures, increases in fH are insufficient to provide all of the additional O(2) required and there are also significant increases in the sO(2) pulses. This creates the situation whereby the same fH at the two temperatures can represent different values of sVO2. PMID- 12077169 TI - Breaking a paradigm: male-produced aggregation pheromone for the Colorado potato beetle. AB - A male-produced aggregation pheromone was identified for the Colorado potato beetle Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). While male beetles produced only minor amounts of the pheromone, its production could be enhanced by topical application of juvenile hormone III (JH III) (eightfold), by antennectomy (40-fold) or by the combined treatment of JH III and antennectomy (almost 200-fold); this enhancement enabled the identification of the compound as (S)-3,7-dimethyl-2-oxo-oct-6-ene-1,3-diol [(S)-CPB I], a unique structure for an insect pheromone. Antennal receptors of both sexes responded selectively to the (S)-enantiomer. Both male and female Colorado potato beetles were attracted to serial source loads of (S)-CPB I in laboratory bioassays; (R)-CPB I was inactive or inhibitory, as demonstrated by the inactivity of the racemate. This is the first identification of a pheromone for the Colorado potato beetle and differs from the paradigm of a female-produced pheromone for this insect. The attractant is also the first male-produced pheromone identified for the Chrysomelidae. The discovery that both JH III and antennectomy increase levels of the pheromone (S) CPB I indicates the existence of a feedback system involving antennal input, and this system may be under hormonal control. PMID- 12077170 TI - Analysis of Ca(2+) uptake into the smooth endoplasmic reticulum of permeabilised sternal epithelial cells during the moulting cycle of the terrestrial isopod Porcellio scaber. AB - In terrestrial isopods, large amounts of Ca(2+) are transported across anterior sternal epithelial cells during moult-related deposition and resorption of CaCO(3) deposits. Because of its toxicity and function as a second messenger, resting cytosolic Ca(2+) levels must be maintained below critical concentrations during epithelial Ca(2+) transport, raising the possibility that organelles play a role during Ca(2+) transit. We therefore studied the uptake of Ca(2+) into Ca(2+)-sequestering organelles by monitoring the formation of birefringent calcium oxalate crystals in permeabilised anterior and posterior sternal epithelium cells of Porcellio scaber during Ca(2+)-transporting and non transporting stages of the moulting cycle using polarised-light microscopy. The results indicate ATP-dependent uptake of Ca(2+) into organelles. Half-maximal crystal growth at a Ca(2+) activity, a(Ca), of 0.4 micromol l(-1) and blockade by cyclopiazonic acid suggest Ca(2+) uptake into the smooth endoplasmic reticulum by the smooth endoplasmic reticulum Ca(2+)-ATPase. Analytical electron microscopical techniques support this interpretation by revealing the accumulation of Ca(2+) containing crystals in smooth membranous intracellular compartments. A comparison of different moulting stages demonstrated a virtual lack of crystal formation in the early premoult stage and a significant fivefold increase between mid premoult and the Ca(2+)-transporting stages of late premoult and intramoult. These results suggest a contribution of the smooth endoplasmic reticulum as a transient Ca(2+) store during intracellular Ca(2+) transit. PMID- 12077171 TI - Electrophysiological properties of the tongue epithelium of the toad Bufo marinus. AB - The dorsal lingual epithelium from the tongue of the toad Bufo marinus was mounted in an Ussing-type chamber, and the short-circuit current (I(sc)) was measured using a low-noise voltage clamp. With NaCl Ringer bathing the mucosal and serosal surfaces of the isolated tissue, an outwardly directed (mucosa positive) I(sc) was measured that averaged -10.71+/-0.82 microA cm(-2) (mean +/- S.E.M., N=24) with a resistance of 615+/-152 Omega cm(2) (mean +/- S.E.M., N=10). Substitution of chloride with sulfate as the anion produced no significant change in I(sc). Fluctuation analysis with either NaCl or Na(2)SO(4) Ringer bathing both sides of the tissue revealed a spontaneous Lorentzian component, suggesting that the I(sc) was the result of K(+) secretion through spontaneously fluctuating channels in the apical membrane of the epithelium. This hypothesis was supported by the reversible inhibition of I(sc) by Ba(2+) added to the mucosal Ringer. Analysis of the kinetics of Ba(2+) inhibition of I(sc) indicates that there might be more than one type of K(+) channel carrying the I(sc). This hypothesis was supported by power spectra obtained with a serosa-to-mucosa K(+) gradient, which could be fitted to two Lorentzian components. At present, the K(+) secretory current cannot be localized to taste cells or other cells that might be associated with the secretion of saliva or mucus. Nonetheless, the resulting increase in [K(+)] in fluid bathing the mucosal surface of the tongue could presumably affect the sensitivity of the taste cells. These results contrast with those from the mammalian tongue, in which a mucosa-negative I(sc) results from amiloride-sensitive Na(+) transport. PMID- 12077172 TI - Hypaxial muscle activity during running and breathing in dogs. AB - The axial muscles of terrestrial vertebrates serve two potentially conflicting functions, locomotion and lung ventilation. To differentiate the locomotor and ventilatory functions of the hypaxial muscles in mammals, we examined the locomotor and ventilatory activity of the trunk muscles of trotting dogs under two conditions: when the ventilatory cycle and the locomotor cycle were coupled and when they were uncoupled. Patterns of muscle-activity entrainment with locomotor and ventilatory events revealed (i) that the internal and external abdominal oblique muscles performed primarily locomotor functions during running yet their activity was entrained to expiration when the dogs were standing, (ii) that the internal and external intercostal, external oblique thoracic and transversus abdominis muscles performed both locomotor and respiratory functions simultaneously, (iii) that the parasternal internal intercostal muscle performed a primarily respiratory function (inspiration) and (iv) that the deep pectoralis and longissimus dorsi muscles performed only locomotor functions and were not active while the dogs were standing still. We conclude that the dual function of many hypaxial muscles may produce functional conflicts during running. The redundancy and complexity of the respiratory musculature as well as the particular pattern of respiratory-locomotor coupling in quadrupedal mammals may circumvent these conflicts or minimize their impact on respiration. PMID- 12077173 TI - ATP as a putative sensory mediator: activation of intrinsic sensory neurons of the myenteric plexus via P2X receptors. AB - The mucosal terminals of sensory neurons intrinsic to the wall of the intestine are sensitive to the chemical environment within the lumen. Lumenal stimuli probably release sensory mediators from the mucosal epithelium, which then activate the nerve terminals indirectly. Here, we tested the idea that ATP activates intrinsic sensory nerve terminals in a way consistent with its being a sensory mediator. We made intracellular recordings from intrinsic sensory neurons located in the myenteric plexus [identified as AH neurons, which are neurons with a long-lasting afterhyperpolarization following the action potential (AP)], located within 1 mm of intact mucosa. Focal electrical stimulation of the mucosa was used to locate and map regions innervated by each neuron. Application of ATP (1-2 mm in the pressure pipette) to these regions elicited trains of APs that originated at the sensory terminals. ATP-gamma-S produced a similar response, but alpha,beta-methylene ATP and 2-methylthio-ATP were only weakly active. The P2 receptor antagonist pyridoxalphosphate-6-azophenyl-2',5'-disulphonic acid (PPADS) (60 microm in the bath) abolished the APs evoked by ATP and ATP-gamma-S but spared similar responses evoked by 5-hydroxytryptamine (5-HT). Another P2 receptor antagonist suramin (100 microm in the bath) did not significantly change the number of APs evoked by ATP. Either ATP or alpha,beta-methylene ATP desensitized the ATP-evoked APs; 50% recovery occurred after approximately 5 sec. The number of APs evoked by ATP was reduced, but not abolished, by the selective 5-HT3 receptor antagonist granisetron (1 microm in the bath). ATP was applied to the cell bodies of sensory neurons to investigate whether the cell bodies express the same P2X receptor as the terminals. ATP evoked a fast depolarization associated with a reduction in input resistance and a reversal potential of -11 mV. This depolarization was potentiated by suramin and blocked by PPADS. We conclude that activation of an atypical excitatory P2X receptor by ATP triggers AP generation in the mucosal processes of the sensory neurons; endogenous 5-HT release may also contribute to activation of the nerve terminals. A similar P2X receptor exists on the cell body of the sensory neuron. Together, these data are consistent with a role for ATP as a sensory mediator in gastrointestinal chemosensory transduction. PMID- 12077174 TI - Increase in the pool size of releasable synaptic vesicles by the activation of protein kinase C in goldfish retinal bipolar cells. AB - Secretion from neurons and neuroendocrine cells is enhanced by the activation of protein kinase C (PKC) in various preparations. We have already reported that transmitter (glutamate) release from Mb1 bipolar cells in the goldfish retina is potentiated by the activation of PKC. However, it is not yet settled whether the potentiation is ascribed to the increase in the pool size of releasable synaptic vesicles or in release probability. In the present study, Ca2+ influx and exocytosis were simultaneously monitored by measuring the presynaptic Ca2+ current and membrane capacitance changes, respectively, in a terminal detached from the bipolar cell. The double pulse protocol was used to estimate separately the changes in the pool size and release probability. The activation of PKC by phorbol 12-myristate 13-acetate (PMA) specifically increased the pool size but not the release probability. PKC was activated by PMA even after the Ca2+ influx was blocked by Co2+. In bipolar cells the releasable pool can be divided into two components: one is small and rapidly exhausted, and the other is large and slowly exocytosed. To identify which component is responsible for the increase in the pool size, the effects of PMA and a PKC-specific inhibitor, bisindolylmaleimide I (BIS), on each component were examined. The slow component was selectively increased by PMA and reduced by BIS. Thus, we conclude that the activation of PKC in Mb1 bipolar cells potentiates glutamate release by increasing the pool size of the slow component. PMID- 12077175 TI - Episodic ataxia type 1 mutations in the human Kv1.1 potassium channel alter hKvbeta 1-induced N-type inactivation. AB - Episodic ataxia type 1 (EA1) is an autosomal dominant neurological disorder affecting both central and peripheral nerve function, causing attacks of imbalance and uncontrolled movements. Genetic linkage studies have identified mutations in the gene encoding the voltage-gated delayed rectifier potassium channel Kv1.1 as underlying EA1. The EA1 mutations E325D and V408A, residing near the cytoplasmic ends of S5 and S6, respectively, induce an unstable open state, resulting in an approximately 10-fold increase in deactivation rates compared with wild-type (WT) channels. Coexpression of EA1 mutations with human Kvbeta1 (hKvbeta1) subunits in Xenopus oocytes yielded channels with altered rapid N-type inactivation. Compared with WT channels, inactivation was approximately twofold slower for homomeric E325D or V408A channels and 1.5-fold slower for heteromeric channels composed of two WT and two E325D or V408A subunits. Recovery from inactivation was approximately 10-fold faster for homomeric E325D or V408A channels and threefold to fourfold faster for heteromeric WT and E325D or V408A channels compared with WT channels. Currents during successive pulses 3 msec in duration given at a rate of 40 kHz decayed e-fold in approximately four pulses for homomeric E325D or V408A and approximately 2.5 pulses for heteromeric channels compared with approximately one pulse for WT channels. These results show that channels containing E325D or V408A subunits, which destabilize the open state, increase the rate of recovery from inactivation. The slower onset and more rapid recovery of hKvbeta1-induced inactivation in channels containing these EA1 subunits may affect temporal integration of action potential firing rates. PMID- 12077176 TI - Coordinate release of ATP and GABA at in vitro synapses of lateral hypothalamic neurons. AB - Autonomic and limbic information is integrated within the lateral hypothalamus (LH), and excitability of LH neurons is important in the control of feeding and behavioral arousal. Despite the prominent expression of P2X-type ATP receptors throughout the hypothalamus, the role of ATP in LH excitability is not known. Perforated-patch-clamp recordings of synaptically coupled neurons from both embryonic chick and postnatal mouse lateral hypothalamus in vitro reveal robust stimulus-evoked purinergic synaptic transmission. Suprathreshold activation elicits reliable and concurrent release of ATP with GABA. Tetrodotoxin-resistant P2X receptor-mediated events are readily observed at LH synapses from the embryonic chick, whereas GABA miniature postsynaptic currents (mPSCs) are recorded in innervated LH neurons from either embryonic chicks or postnatal mice. Two distinct mPSCs are recorded at ATP-GABA cosynapses; one has a monoexponential decay phase and is modulated by flunitrazepam, and the other has a decay phase that is best fit by a sum of two exponential functions (tau(fast) and tau(slow)), and only the tau(slow) component is affected by flunitrazepam. Bicuculline does not completely inhibit all mPSCs. The remaining bicuculline-resistant mPSCs are blocked by suramin, and their decay phase is briefer than that of GABAergic mPSCs. Furthermore, at a holding potential intermediate for the reversal potentials of GABA(A) and P2X receptors, little or no current is observed, consistent with concomitant release (and detection) of GABA and ATP. Together, our data suggest that a subset of spontaneous and evoked PSCs arise from the concurrent activation of both GABA(A) and P2X receptors. PMID- 12077177 TI - GABAergic terminals are required for postsynaptic clustering of dystrophin but not of GABA(A) receptors and gephyrin. AB - In rat hippocampal cultures, we show by multilabeling immunocytochemistry that pyramidal cells, which receive little or no GABAergic input, mistarget alpha2 GABA(A) receptors and gephyrin to glutamatergic terminals. This mismatch does not occur in neurons innervated by numerous GABAergic terminals. A similar phenomenon has been reported for isolated autaptic hippocampal neurons (Rao et al., 2000). GABAergic synapses typically form multiple release sites apposed to GABA(A) receptor and gephyrin clusters. Remarkably, dystrophin, a protein highly abundant in skeletal muscle membranes, is extensively colocalized with alpha2-GABA(A) receptors exclusively opposite GABAergic terminals. In addition, selective apposition of syntrophin and beta-dystroglycan to GABAergic presynaptic terminals suggests that the entire dystrophin-associated protein complex (DPC) clusters at GABAergic synapses. In contrast to gephyrin and GABA(A) receptors, DPC proteins are not mistargeted to glutamatergic synapses, indicating independent clustering mechanisms. This was confirmed in hippocampal neurons cultured from GABA(A) receptor gamma2 subunit-deficient mice. Clustering of GABA(A) receptor and gephyrin in these neurons was strongly impaired, whereas clustering of dystrophin and associated proteins was unaffected by the absence of the gamma2 subunit. Our results indicate that accumulation of dystrophin and DPC proteins at GABAergic synapses occurs independently of postsynaptic GABA(A) receptors and gephyrin. We suggest that selective signaling from GABAergic terminals contributes to postsynaptic clustering of dystrophin. PMID- 12077178 TI - P2X receptor trafficking in neurons is subunit specific. AB - P2X receptors within the CNS mediate excitatory synaptic transmission and also act presynaptically to modulate neurotransmitter release. We have studied the targeting and trafficking of P2X4 and P2X2 receptors heterologously expressed in cultured olfactory bulb neurons. Homomeric P2X4 receptors had a punctate distribution, and many of the puncta colocalized with early endosomes. In contrast, P2X2 receptors were primarily localized at the plasma membrane. By antibody-labeling of surface receptors in living neurons, we showed that P2X4 receptors undergo rapid constitutive internalization and subsequent reinsertion into the plasma membrane, whereas P2X2 receptors were not regulated in such a way. The internalization of P2X4 receptors was dynamin-dependent, and the binding of ATP enhanced the basal rate of retrieval in a Ca2+-independent manner. The presence of the P2X4 subunit in a P2X4/6 heteromer governed the trafficking properties of the receptor. P2X receptors acted presynaptically to enhance the release of glutamate, suggesting that the regulated cycling of P2X4-containing receptors might provide a mechanism for modulation of synaptic transmission. PMID- 12077179 TI - Accumulation of SOD1 mutants in postnatal motoneurons does not cause motoneuron pathology or motoneuron disease. AB - Transgenic mice expressing high levels of familial amyotrophic lateral sclerosis (FALS)-associated mutant superoxide dismutase 1 (SOD1) under the control of a human SOD1 minigene (hMg) accumulate mutant protein ubiquitously and develop motoneuron disease. However, restricted expression of SOD1 mutants in neurons apparently does not cause motor impairments in mice. Here, we investigated the possible pathogenic roles of mutant SOD1 accumulation in motoneurons. First, we used a Thy1 expression cassette to drive high constitutive expression of transgene in postnatal mouse neurons, including upper and lower motoneurons. Second, we expressed human (h) SOD1(G93A) and hSOD1(G85R) as transgenes (i.e., two SOD1 mutants with aggressive pathogenic properties in inducing FALS). Third, in addition to clinical signs of disease, we monitored early signs of disease onset and pathogenesis, including muscle innervation, astrogliosis in the spinal cord, and accumulation of ubiquitinated deposits in motoneurons and astrocytes. We report that high-level expression and accumulation of the mutant proteins in neurons failed to produce any detectable sign of pathology or disease in these transgenic mice. Crossing hMg-SOD1(G93A) mice (Gurney et al., 1994) with Thy1 SOD1(G93A) mice produced double-transgenic mice with spinal cord SOD1(G93A) levels that were approximately twofold higher than in the hMg-SOD1(G93A) single transgenics but did not affect the onset or progression of pathology or motoneuron disease. The accumulation of mutant SOD1 in postnatal motoneurons is thus not sufficient and probably also not critical to induce or accelerate motoneuron disease in FALS mice. The pathogenic process in FALS may involve non neuronal cells, and selective vulnerability of motoneurons to this process may lead to motoneuron pathology and disease. PMID- 12077180 TI - A novel function of monomeric amyloid beta-protein serving as an antioxidant molecule against metal-induced oxidative damage. AB - Aggregated and oligomeric amyloid beta-protein (Abeta) is known to exhibit neurotoxicity. However, the action of Abeta monomers on neurons is not fully understood. We have studied aggregation state-dependent actions of Abeta and found an oligomer-specific effect of Abeta on lipid metabolism in neurons (Michikawa et al., 2001). Here, we show a novel function of monomeric Abeta1-40, which is the major species found in physiological fluid, as a natural antioxidant molecule that prevents neuronal death caused by transition metal-induced oxidative damage. Monomeric Abeta1-40, which is demonstrated by SDS-PAGE after treatment with glutaraldehyde, protects neurons cultured in a medium containing 1.5 microm Fe(II) without antioxidant molecules. Metal ion chelators such as EDTA, CDTA (trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid), and DTPA (diethylenetriamine-N,N,N',N",N"-penta-acetic acid, an iron-binding protein, transferrin, and antioxidant scavengers such as catalase, glutathione, and vitamin E also inhibit neuronal death under the same conditions. Monomeric Abeta1 40 inhibits neuronal death caused by Cu(II), Fe(II), and Fe(III) but does not protect neurons against H2O2-induced damage. Monomeric Abeta1-40 inhibits the reduction of Fe(III) induced by vitamin C and the generation of superoxides and prevents lipid peroxidation induced by Fe(II). Abeta1-42 remaining as a monomer also exhibits antioxidant and neuroprotective effects. In contrast, oligomeric and aggregated Abeta1-40 and Abeta1-42 lose their neuroprotective activity. These results indicate that monomeric Abeta protects neurons by quenching metal inducible oxygen radical generation and thereby inhibiting neurotoxicity. Because aggregated Abeta is known to be an oxygen radical generator, our results provide a novel concept that the aggregation-dependent biological effects of Abeta are dualistic, being either an oxygen radical generator or its inhibitor. PMID- 12077181 TI - Calpain activation in Huntington's disease. AB - Huntington's disease (HD) is a neurodegenerative disorder caused by a CAG expansion that results in elongation of the polyglutamine tract at the N terminus of huntingtin (Htt). Abnormal proteolytic processing of mutant Htt has been implicated as a critical step in the initiation of HD. The protease(s) involved in this process has not been fully characterized. Here we report that activated calpain was detected in the caudate of human HD tissue but not in age-matched controls. In addition, one of the major N-terminal Htt proteolytic fragments found in human HD tissue appears to be derived from calpain cleavage. Htt fragments in HD lysates were similar in size to those produced by exposure of in vitro-translated Htt to exogenous calpain. Incubation of in vitro-translated Htt with calpain generated a cascade of cleavage events with an initial intermediate cleavage product at 72 kDa and a final cleavage product at 47 kDa. The rate of cleavage of Htt by calpain was polyglutamine-length-dependent. These results suggest that cleavage of Htt in human HD tissue is mediated in part by the Ca2+ activated neutral protease, calpain. PMID- 12077182 TI - Control and plasticity of intercellular calcium waves in astrocytes: a modeling approach. AB - Intercellular Ca2+ waves in astrocytes are thought to serve as a pathway of long range signaling. The waves can propagate by the diffusion of molecules through gap junctions and across the extracellular space. In rat striatal astrocytes, the gap-junctional route was shown to be dominant. To analyze the interplay of the processes involved in wave propagation, a mathematical model of this system has been developed. The kinetic description of Ca2+ signaling within a single cell accounts for inositol 1,4,5-trisphosphate (IP3) generation, including its activation by cytoplasmic Ca2+, IP3-induced Ca2+ liberation from intracellular stores and various other Ca2+ transports, and cytoplasmic diffusion of IP3 and Ca2+. When cells are coupled by gap junction channels in a two-dimensional array, IP3 generation in one cell triggers Ca2+ waves propagating across some tens of cells. The spatial range of wave propagation is limited, yet depends sensitively on the Ca2+-mediated regeneration of the IP3 signal. Accordingly, the term "limited regenerative signaling" is proposed. The gap-junctional permeability for IP3 is the crucial permissive factor for wave propagation, and heterogeneity of gap-junctional coupling yields preferential pathways of wave propagation. Processes involved in both signal initiation (activation of IP3 production caused by receptor agonist) and regeneration (activation of IP3 production by Ca2+, loading of the Ca2+ stores) are found to exert the main control on the wave range. The refractory period of signaling strongly depends on the refilling kinetics of the Ca2+ stores. Thus the model identifies multiple steps that may be involved in the regulation of this intercellular signaling pathway. PMID- 12077183 TI - Protein kinase modulation of dendritic K+ channels in hippocampus involves a mitogen-activated protein kinase pathway. AB - We investigated mitogen-activated protein kinase (MAPK) modulation of dendritic, A-type K+ channels in CA1 pyramidal neurons in the hippocampus. Activation of cAMP-dependent protein kinase A (PKA) and protein kinase C (PKC) leads to an increase in the amplitude of backpropagating action potentials in distal dendrites through downregulation of transient K+ channels in CA1 pyramidal neurons in the hippocampus. We show here that both of these signaling pathways converge on extracellular-regulated kinases (ERK)-specific MAPK in mediating this reduction in dendritic K+ current, which is confirmed, in parallel, by biochemical assays using phosphospecific antibodies against the ppERK and pKv4.2. Furthermore, immunostaining indicates dendritic localization of ppERK and pKv4.2. Taken together, these results demonstrate that dendritic, A-type K+ channels are dually regulated by PKA and PKC through a common downstream pathway involving MAPK, and the modulation of these K+ channels may be accounted for by the phosphorylation of Kv4.2 subunits. PMID- 12077184 TI - Cyclin-dependent kinase 5 phosphorylates disabled 1 independently of Reelin signaling. AB - Two major signaling pathways that control neuronal positioning during brain development have been uncovered as a result of genetic and biochemical studies on neurological mouse mutants. Mice deficient in Reelin, Disabled 1 (Dab1), or both the very low-density lipoprotein receptor (VLDLR) and the apolipoprotein E receptor 2 (ApoER2) exhibit identical neuroanatomic defects in laminar structures throughout the brain. These proteins function as components of the Reelin signaling pathway. Reelin is a secreted glycoprotein that binds to VLDLR and ApoER2, inducing tyrosine phosphorylation of Dab1, an intracellular adapter protein. Neuronal migration is also regulated by cyclin-dependent kinase 5 (Cdk5) and its activating subunits p35 and p39. Mice deficient in Cdk5, p35, or both p35 and p39 exhibit lamination defects that are similar but not identical to those observed in mice with a defect in the Reelin signaling pathway. Cdk5 phosphorylates proteins that maintain cytoskeletal structures and promote cell motility. To explore the possibility that Cdk5 influences the Reelin pathway, we sought to determine whether Dab1 is a substrate for Cdk5. Here we show that Cdk5 phosphorylates Dab1 on serine 491 in vitro and in vivo, independently of Reelin signaling. We also show that ectopic neurons in Cdk5-deficient mice exhibit reduced levels of Reelin signaling during later stages of cortical development, although Cdk5 is not required for Reelin-induced tyrosine phosphorylation of Dab1. Although the functional significance of Dab1 serine phosphorylation is unclear, our results suggest that there is biochemical cross-talk between two signaling pathways that control cell positioning. PMID- 12077185 TI - Light response of retinal ON bipolar cells requires a specific splice variant of Galpha(o). AB - Glutamate released onto retinal ON bipolar neurons binds to a metabotropic receptor to activate a heterotrimeric G-protein (G(o)) that ultimately closes a nonspecific cation channel. Signaling requires the alpha subunit (Galpha(o)), but its effector is unknown. Because Galpha(o) is transcribed into two splice variants (alpha(o1) and alpha(o2)) that differ in the key GTPase domain, the next step in elucidating this pathway was to determine which splice variant carries the signal. Here we show by reverse transcription-PCR and Western blots that retina expresses both splice variants. Furthermore, in situ hybridization and immunostaining on mouse retina deficient in one splice variant or the other show that both alpha(o1) and alpha(o2) are expressed by ON bipolar cells but that alpha(o1) is much more abundant. Finally, electroretinography performed on mice deficient for one splice variant or the other shows that the positive b-wave (response of ON bipolar cells to rod and cone input) requires alpha(o1) but not alpha(o2). Thus, the light response of the ON bipolar cell is probably carried by its strongly expressed splice variant, Galpha(o1). PMID- 12077186 TI - Perineuronal oligodendrocytes protect against neuronal apoptosis through the production of lipocalin-type prostaglandin D synthase in a genetic demyelinating model. AB - The genetic demyelinating mouse "twitcher" is a model of the human globoid cell leukodystrophy, caused by galactosylceramidase (GALC) deficiency. Demyelination in the twitcher brain is secondary to apoptotic death of oligodendrocytes (OLs). Lipocalin-type prostaglandin (PG) D synthase (L-PGDS), a protein expressed in mature OLs, was progressively upregulated in twitcher OLs; whereas expression of OL-associated proteins such as carbonic anhydrase II, myelin basic protein, and myelin-associated glycoprotein was downregulated during demyelination in twitcher brains. The upregulation of L-PGDS was more remarkable in perineuronal OLs than in interfascicular OLs. A larger number of L-PGDS-positive OLs was found in selected fiber tracts of twitcher brains where fewer apoptotic cells were detected. The distribution of L-PGDS-positive OLs was inversely related to the severity of demyelination, as assessed by accumulation of scavenger macrophages. Mice doubly deficient for L-PGDS and GALC disclosed a large number of apoptotic neurons, which were never seen in twitcher brains, in addition to an increased number of apoptotic OLs. A linear positive correlation was observed between the population of L-PGDS-positive OLs in the twitcher brain and the ratio of apoptotic nuclei in the double mutant versus those in the twitcher, suggesting a dose-dependent effect of L-PGDS against apoptosis. These lines of evidence suggest that L-PGDS is an anti-apoptotic molecule protecting neurons and OLs from apoptosis in the twitcher mouse. This is a novel example of OL-neuronal interaction. PMID- 12077187 TI - Polyglutamine-expanded ataxin-7 promotes non-cell-autonomous purkinje cell degeneration and displays proteolytic cleavage in ataxic transgenic mice. AB - Spinocerebellar ataxia (SCA) type 7 is an inherited neurodegenerative disorder caused by expansion of a polyglutamine tract within the ataxin-7 protein. To determine the molecular basis of polyglutamine neurotoxicity in this and other related disorders, we produced SCA7 transgenic mice that express ataxin-7 with 24 or 92 glutamines in all neurons of the CNS, except for Purkinje cells. Transgenic mice expressing ataxin-7 with 92 glutamines (92Q) developed a dramatic neurological phenotype presenting as a gait ataxia and culminating in premature death. Despite the absence of expression of polyglutamine-expanded ataxin-7 in Purkinje cells, we documented severe Purkinje cell degeneration in 92Q SCA7 transgenic mice. We also detected an N-terminal truncation fragment of ataxin-7 in transgenic mice and in SCA7 patient material with both anti-ataxin-7 and anti polyglutamine specific antibodies. The appearance of truncated ataxin-7 in nuclear aggregates correlates with the onset of a disease phenotype in the SCA7 mice, suggesting that nuclear localization and proteolytic cleavage may be important features of SCA7 pathogenesis. The non-cell-autonomous nature of the Purkinje cell degeneration in our SCA7 mouse model indicates that polyglutamine induced dysfunction in adjacent or connecting cell types contributes to the neurodegeneration. PMID- 12077188 TI - Dendritic remodeling and growth of motoneurons during metamorphosis of Drosophila melanogaster. AB - Insect motoneurons display dramatic dendritic plasticity during metamorphosis. Many larval motoneurons survive to adulthood but undergo dendritic regression and outgrowth as they are incorporated into developing circuits. This study explores the dendritic remodeling and development of Drosophila motoneurons MN1-MN5, which innervate indirect flight muscles of the adult. MN1-MN5 are persistent larval neurons exhibiting two distinct metamorphic histories. MN1-MN4 are born in the embryo, innervate larval muscles, and undergo dendritic regression and regrowth during metamorphosis. MN5, which was identified through a combination of intracellular dye injection and retrograde staining at all stages, is also born embryonically but remains developmentally arrested until the onset of metamorphosis. In the larva, MN5 lacks dendrites, and its axon stops in the mesothoracic nerve without innervating a target muscle. It is dye coupled to the peripherally synapsing interneuron, which will become part of the giant fiber escape circuit of the adult fly. During pupal development, MN5 undergoes de novo dendritic growth and extension of its axon to innervate the developing target muscle. Its unique developmental history and identifiability make MN5 well suited for the study of dendritic growth using genetic and neurophysiological approaches. PMID- 12077189 TI - The neural cell adhesion molecule L1 potentiates integrin-dependent cell migration to extracellular matrix proteins. AB - The L1 adhesion molecule regulates axon growth and is mutated in the X-linked mental retardation syndrome CRASH (acronym for corpus callosum agenesis, retardation, aphasia, spastic paraplegia, hydrocephalus). A novel role for L1 as a potentiator of neuronal cell migration to extracellular matrix proteins through beta1 integrins and intracellular signaling to mitogen-activated protein (MAP) kinase was identified. L1 potentiated haptotactic migration of B35 neuroblastoma cells toward fibronectin, vitronectin, and laminin through the signaling intermediates c-Src, phosphatidylinositol-3 kinase, and MAP kinase. L1 potentiated migration toward fibronectin through alpha5beta1 integrin in human embryonic kidney 293 cells and depended on determinants of L1 endocytosis: dynamin I, c-Src, and the AP2/clathrin binding site (Arg-Ser-Leu-Glu) in the neuronal splice form of L1. L1 clustering on the cell surface enhanced the internalization of activated beta1 integrins and L1 into distinct endocytic vesicles. L1-potentiated migration, enhancement of beta1 integrin endocytosis, and activation of MAP kinase were coordinately inhibited by mutation of an RGD sequence in the sixth immunoglobulin-like domain of L1. Moreover, three CRASH mutations in the L1 cytoplasmic domain (1194L, S1224L, Y1229H), two of which interfere with ankyrin association, inhibited L1-potentiated migration and MAP kinase activation. Function-blocking antibodies to L1 and beta1 integrin retarded the migration of 5-bromo-2'-deoxyuridine-labeled mouse cerebellar granule cells in slice cultures, underscoring the potential physiological relevance of these findings. These studies suggest that L1 functionally interacts with beta1 integrins to potentiate neuronal migration toward extracellular matrix proteins through endocytosis and MAP kinase signaling, and that impairment of this function by L1 cytoplasmic domain mutations may contribute to neurological deficits in CRASH. PMID- 12077190 TI - Growth cone collapse induced by semaphorin 3A requires 12/15-lipoxygenase. AB - Detection of a repellent factor, such as a semaphorin (Sema), causes localized collapse of the growth cone and directs the neurite away from the repellent. Growth cone collapse results from concomitant cytoskeletal rearrangements and detachment of adhesion sites from the extracellular matrix, via mostly unknown signaling mechanisms. In cultures of dorsal root ganglion neurons, we found that Sema3A treatment stimulates the synthesis of the eicosanoid, 12(S) hydroxyeicosatetraenoic acid (HETE), whereas Sema3A-induced growth cone collapse is prevented when 12(S)-HETE synthesis is blocked with an inhibitor of 12/15 lipoxygenase (LO). Exogenously applied product of 12/15-LO, 12(S)-HETE, mimics Sema3A-induced collapse. As observed by interference reflection and confocal microscopy, 12(S)-HETE causes the loss of growth cone adhesion sites. The adhesion site effect seems partially independent of the actin cytoskeleton because growth cones treated with Sema3A and 12/15-LO inhibitor remain spread despite actin cytoskeleton loss. These studies demonstrate that 12/15-LO activity is a necessary step in Sema3A collapse signaling in growth cones and suggest a mechanism for its action. PMID- 12077191 TI - Lentivirally delivered glial cell line-derived neurotrophic factor increases the number of striatal dopaminergic neurons in primate models of nigrostriatal degeneration. AB - The primate striatum contains tyrosine hydroxylase (TH)-immunoreactive (ir) neurons, the numbers of which are augmented after dopamine depletion. Glial cell line-derived neurotrophic factor (GDNF) strongly modulates the viability and phenotypic expression of dopamine ventral mesencephalic neurons. The effect of GDNF on TH-ir neurons intrinsic to the striatum has yet to be investigated. In the present study, stereological counts of TH-ir striatal neurons in aged and parkinsonian nonhuman primates revealed that GDNF delivered via a lentiviral vector (lenti-) further increased the number of these cells. Aged monkeys treated with lenti-GDNF displayed an eightfold increase in TH-ir neurons relative to lenti-beta-galactosidase-treated monkeys. Unilateral 1-methyl-4-phenyl- 1,2,3,6 tetrahydropyridine treatment alone in young monkeys resulted in a bilateral eightfold increase in TH-ir striatal cells. This effect was further magnified sevenfold on the side of lenti-GDNF treatment. These cells colocalized with the neuronal marker neuronal-specific nuclear protein. Some of these cells colocalized with GDNF-ir, indicating that an alteration in phenotype may occur by the direct actions of this trophic factor. Thus, GDNF may mediate plasticity in the dopamine-depleted primate brain, which may serve to compensate for cell loss by converting striatal neurons to a dopaminergic phenotype. PMID- 12077192 TI - Axonally transported peripheral signals regulate alpha-internexin expression in regenerating motoneurons. AB - The class IV neuronal intermediate filament (IF) family proteins includes the neurofilament (NF) triplet proteins NF-L, NF-M, and NF-H and also the more recently characterized alpha-internexin-NF66. It is well established that NF-L, M, and -H protein and mRNA are downregulated after peripheral nerve injury. We examined alpha-internexin protein expression after three facial nerve lesion paradigms: crush, transection, and resection. Alpha-internexin immunoreactivity was absent in the perikarya of uninjured facial motoneurons but increased dramatically in all three injury paradigms, with maximum immunoreactivity observed at 7 d after injury. Twenty-eight days after nerve crush or transection, there was a dramatic decrease in the number of alpha-internexin-positive cells. In contrast, alpha-internexin remained elevated 28 d after nerve resection, an injury that hinders regeneration and target reinnervation. In situ hybridization studies showed an increase in alpha-internexin mRNA expression in the facial nucleus at 7 and 14 d after injury. Retrograde transport of fluorogold from the whisker pads to the facial nucleus was seen only in motoneurons that lacked alpha internexin immunoreactivity, supporting the idea that target reinnervation and inhibitory signals from the periphery regulate the expression of alpha internexin. Blockage of axonal transport through local colchicine application induced strong immunoreactivity in motoneurons. Alpha-internexin expression was also examined after central axotomy of rubrospinal neurons, which constitutively show alpha-internexin immunoreactivity. After rubrospinal tractotomy, alpha internexin immunoreactivity transiently increased by 7 d after injury but returned to control levels by 14 d. We conclude that alpha-internexin upregulation in injured motoneurons suggests a role for this IF protein in neuronal regeneration. PMID- 12077193 TI - Regulation by protein kinase A switching of axonal pathfinding of zebrafish olfactory sensory neurons through the olfactory placode-olfactory bulb boundary. AB - Cumulative evidence suggests that neural network formation requires an ingenious regulation of the attractive and repulsive responses of growing axons to guidance cues. We examined the role of intracellular protein kinase A (PKA) signaling in the axonal pathfinding of olfactory sensory neurons in transparent zebrafish embryos. Microinjection of an olfactory marker protein gene promoter-driven double-cassette vector directed the expression of both the dominant form of PKA and green fluorescent protein fused with the microtubule-associated protein tau in the same olfactory neurons. The dominant-negative form of PKA enhanced the turning of olfactory neuron axons in the olfactory placode, whereas the disturbance effect of the constitutively active form on the axonal pathfinding was prominent in the olfactory bulb. Consistently, forskolin treatment severely inhibited the axonal extension in the olfactory bulb, but not in the olfactory placode. These results suggest that the switching of PKA signaling in developing olfactory sensory neurons is important for axonal pathfinding through the boundary between the olfactory placode and the olfactory bulb in vivo. We thus propose that the regulation of PKA signaling plays a key role in the long distance axonal pathfinding through intermediate guideposts. PMID- 12077194 TI - Expression of p73 and Reelin in the developing human cortex. AB - Cajal-Retzius (CR) cells of the developing neocortex secrete Reelin (Reln), a glycoprotein involved in neuronal migration. CR cells selectively express p73, a p53 family member implicated in cell survival and apoptosis. Immunocytochemistry in prenatal human telencephalon reveals a complex sequence of migration waves of p73- and Reln-immunoreactive (IR) neurons into the cortical marginal zone (MZ). At early preplate stages, p73/Reln-IR cells arise in distinct sectors of the telencephalon, including cortical primordium and ganglionic eminences. After the appearance of the cortical plate, further p73/Reln-IR cells originate in the medial periolfactory forebrain. In addition, p73 marks a novel cell population that appears at the choroid-cortical junction or cortical hem before the emergence of the dorsal hippocampus. A pronounced mediolateral gradient in the density of p73/Reln-IR neurons in the neocortical MZ at 8 gestational weeks suggests that a subset of CR cells migrate tangentially from cortical hem and taenia tecta into neocortical territory. This hypothesis is supported by the absence of p73-transcripts in prospective neocortex of p73-/-mice at embryonic day 12 (E12), whereas they are present in cortical hem and taenia tecta. In the p73-/- preplate, Reln is faintly expressed in a calretinin-positive cell population, not present in this form in the E12 wild-type cortex. At P2, Reln-IR CR cells are undetectable in the p73-/- cortex, whereas Reln-expression in interneurons is unchanged. Our results point to a close association between p73 and Reln in CR cells of the developing neocortex, with a partial dissociation in early preplate and basal telencephalon, and to a p73-mediated role of the cortical hem in neocortical development. PMID- 12077195 TI - Interactions between TrkB signaling and serotonin excess in the developing murine somatosensory cortex: a role in tangential and radial organization of thalamocortical axons. AB - Mice lacking monoamine oxidase A (MAOA) display high levels of brain serotonin during the first postnatal week, causing an exuberant outgrowth of thalamocortical axons (TCAs) in layer IV of the somatosensory cortex (S1). We asked whether this exuberance is attributable to abnormal TrkB signaling, because modulation of TrkB signaling during a critical period dramatically influences the segregation and the morphology of TCAs in layer IV of the visual cortex. Using in situ hybridization and ELISA immunoassays, we showed that the levels of trkB mRNA and BDNF and neurotrophin-4 (NT-4) proteins are normal in the thalamus and the cortex of mice lacking MAOA during barrel field formation. Because the release of BDNF and NT-4 could be abnormal in MAOA knock-out (KO) mice, we tested whether abnormal TrkB signaling is required for TCA exuberance in MAOA-KO mice by generating mice lacking both trkB and MAOA. Surprisingly, these mice exhibited more severe phenotypes than those found in MAOA-KO mice: a widespread tangential expansion of TCAs in layer IV of the cortex, resulting in a fusion of all sensory representations and a radial expansion of TCAs in layers II-III of the cortex. Careful examination of mice lacking trkB alone revealed subtle alterations of TCAs, with abnormal invasion of layer III. This study reveals the following: (1) expression of trkB, BDNF, and NT-4 are not modulated by an excess of serotonin during barrel formation, (2) TrkB signaling limits branching of TCAs in inappropriate supragranular cortical layers, and (3) serotonin and TrkB signaling act together to cluster thalamocortical axons in layer IV. PMID- 12077197 TI - Transient PKA activity is required for initiation but not maintenance of BDNF mediated protection from nitric oxide-induced growth-cone collapse. AB - Growing axons during development are guided to their targets by the activity of their growth cones. Growth cones integrate positive and negative guidance cues in deciding the direction in which to extend. We demonstrated previously that treatment of embryonic retinal ganglion cells with brain-derived neurotrophic factor (BDNF) protects their growth cones from collapse induced by nitric oxide (NO). BDNF stabilizes growth-cone actin filaments against NO-induced depolymerization. In the present study, we examined the signaling mechanism involved in BDNF-mediated protection. We found that BDNF causes transient activation of protein kinase A (PKA) during the first 5 min of treatment. Treatment with PKA inhibitors before or in conjunction with BDNF treatment blocked the protective effects of BDNF. The effects of BDNF, however, were not blocked when addition of PKA inhibitors was delayed as little as 15 min after BDNF treatment. When cultures raised overnight in BDNF were treated with PKA inhibitors, BDNF-mediated protection did not end, demonstrating that the maintenance of the protective effects of BDNF is independent of PKA activity. The BDNF-induced activation of PKA was required for BDNF-mediated stabilization of growth-cone actin filaments against depolymerization by cytochalasin D. Finally, the initiation and maintenance of the protective effects of BDNF required protein synthesis. Collectively, these data demonstrate that PKA signaling is required only for an early phase of BDNF-mediated protection from NO-induced growth-cone collapse. PMID- 12077198 TI - Chick PTPsigma regulates the targeting of retinal axons within the optic tectum. AB - Chick PTPsigma (cPTPsigma), also known as CRYPalpha, is a receptor-like protein tyrosine phosphatase found on axons and growth cones. Putative ligands for cPTPsigma are distributed within basement membranes and on glial end feet of the retina, optic nerve, and optic tectum, suggesting that cPTPsigma signaling is occurring along the whole retinotectal pathway. We have shown previously that cPTPsigma plays a role in supporting the retinal phase of axon outgrowth. Here we have now addressed the role of cPTPsigma within retinal axons as they undergo growth and topographic targeting in the optic tectum. With the use of retroviruses, a secretable cPTPsigma ectodomain was ectopically expressed in ovo in the developing chick optic tectum, with the aim of directly disrupting the function of endogenous cPTPsigma. In ovo, the secreted ectodomains accumulated at tectal sites in which cPTPsigma ligands are also specifically found, suggesting that they are binding to these endogenous ligands. Anterograde labeling of retinal axons entering these optic tecta revealed abnormal axonal phenotypes. These included the premature stalling and arborization of fibers, excessive pretectal arbor formation, and diffuse termination zones. Most of the defects were rostral of the predicted termination zone, indicating that cPTPsigma function is necessary for sustaining the growth of retinal axons over the optic tectum and for directing axons to their correct sites of termination. This demonstrates that regulation of cPTPsigma signaling in retinal axons is required for their topographic mapping, the first evidence of this function for a receptor like protein tyrosine phosphatase in the retinotectal projection. PMID- 12077196 TI - Alpha7 nicotinic acetylcholine receptors occur at postsynaptic densities of AMPA receptor-positive and -negative excitatory synapses in rat sensory cortex. AB - NMDA receptor (NMDAR) activation requires concurrent membrane depolarization, and glutamatergic synapses lacking AMPA receptors (AMPARs) are often considered "silent" in the absence of another source of membrane depolarization. During the second postnatal week, NMDA currents can be enhanced in rat auditory cortex through activation of the alpha7 nicotinic acetylcholine receptor (alpha7nAChR). Electrophysiological results support a mainly presynaptic role for alpha7nAChR at these synapses. However, immunocytochemical evidence that alpha7nAChR is prevalent at postsynaptic sites of glutamatergic synapses in hippocampus and neocortex, along with emerging electrophysiological evidence for postsynaptic nicotinic currents in neocortex and hippocampus, has prompted speculation that alpha7nAChR allows for activation of NMDAR postsynaptically at synapses lacking AMPAR. Here we used dual immunolabeling and electron microscopy to examine the distribution of alpha7nAChR relative to AMPAR (GluR1, GluR2, and GluR3 subunits combined) at excitatory synapses in somatosensory cortex of adult and 1-week-old rats. alpha7nAChR occurred discretely over most of the thick postsynaptic densities in all cortical layers of both age groups. AMPAR immunoreactivity was also detectable at most synapses; its distribution was independent of that of alpha7nAChR. In both age groups, approximately one-quarter of asymmetrical synapses were alpha7nAChR positive and AMPAR negative. The variability of postsynaptic alpha7nAChR labeling density was greater at postnatal day (PD) 7 than in adulthood, and PD 7 neuropil contained a subset of small AMPA receptor negative synapses with a high density of alpha7nAChR immunoreactivity. These observations support the idea that acetylcholine receptors can aid in activating glutamatergic synapses and work together with AMPA receptors to mediate postsynaptic excitation throughout life. PMID- 12077199 TI - Calcineurin plays different roles in group II metabotropic glutamate receptor- and NMDA receptor-dependent long-term depression. AB - We investigated metabotropic glutamate receptor (mGluR)-dependent long-term depression (LTD) in hippocampal CA1 pyramidal neurons of 6- to 8-d-old [postnatal days 6-8 (P6-P8)] and 21- to 25-d-old (P21-P25) rats. In P6-P8 rats, induction of LTD depended on the activity of group II mGluRs. In P21-P25 rats, however, this LTD disappeared, and instead, NMDA receptor (NMDAR)-dependent LTD appeared. A bath containing a specific calcineurin (CaN) inhibitor restored the group II mGluR-dependent LTD in the neurons of the P21-P25 rats. Although postsynaptic injection of CaN inhibitors suppressed NMDAR-dependent LTD, it did not affect induction of group II mGluR-dependent LTD. These results demonstrate that CaN plays different roles in the induction of two forms of LTD: presynaptic CaN inhibits group II mGluR-dependent LTD, whereas postsynaptic CaN facilitates NMDAR dependent LTD. These findings are the first demonstration in vitro of group II mGluR-dependent LTD that is negatively regulated by CaN via an age-dependent mechanism. PMID- 12077200 TI - D1 dopamine receptor supersensitivity in the dopamine-depleted striatum results from a switch in the regulation of ERK1/2/MAP kinase. AB - Dopamine effects in the striatum are mediated principally through the D1 and D2 dopamine receptor subtypes, which are segregated to the direct and indirect striatal projection neurons. After degeneration of the nigrostriatal dopamine system, direct pathway neurons display a supersensitive response to D1 dopamine receptor agonists, which is demonstrated by the induction of immediate early genes (IEGs), such as c-fos. Here we show, using analysis of receptor-mediated signal transduction, including protein phosphorylation and induction of IEGs, that D1 dopamine receptor supersensitivity is attributable to a switch to ERK1/2/MAP kinase (extracellular signal-regulated kinase/mitogen-activated protein kinase) in direct pathway neurons. Normally, in the dopamine-intact striatum, activation of ERK1/2/MAP kinase is shown to be restricted to indirect and not direct pathway neurons in response to stimulation of corticostriatal afferents. Moreover, in the dopamine-intact striatum, treatment with full D1 dopamine receptor agonists or stimulation of nigrostriatal dopaminergic afferents, both of which result in the induction of IEGs in direct striatal projection neurons, does not activate ERK1/2/MAP kinase. However, after degeneration of the nigrostriatal dopaminergic pathway, ERK1/2/MAP kinase is activated in direct pathway neurons in response to D1 dopamine receptor agonists either alone or when combined with stimulation of corticostriatal afferents. Inhibitors of MEK (MAP kinase kinase), which is responsible for phosphorylation of ERK1/2/MAP kinase, blocks D1 dopamine receptor agonist activation of ERK1/2/MAP kinase in the dopamine-depleted striatum, as well as the supersensitive induction of IEGs. These results demonstrate that dopamine input to the striatum maintains distinct forms of protein kinase-mediated gene regulation in the direct and indirect striatal projection neurons. PMID- 12077201 TI - The spatiotemporal dynamics of illusory contour processing: combined high-density electrical mapping, source analysis, and functional magnetic resonance imaging. AB - Because environmental information is often suboptimal, visual perception must frequently rely on the brain's reconstruction of contours absent from retinal images. Illusory contour (IC) stimuli have been used to investigate these "filling-in" processes. Intracranial recordings and neuroimaging studies show IC sensitivity in lower-tier area V2, and to a lesser extent V1. Some interpret these data as evidence for feedforward processing of IC stimuli, beginning at lower-tier visual areas. On the basis of lesion, visual evoked potentials (VEP), and neuroimaging evidence, others contend that IC sensitivity is a later, higher order process. Whether IC sensitivity seen in lower-tier areas indexes feedforward or feedback processing remains unresolved. In a series of experiments, we addressed the spatiotemporal dynamics of IC processing. Centrally presented IC stimuli resulted in early VEP modulation (88-100 msec) over lateral occipital (LOC) scalp--the IC effect. The IC effect followed visual response onset by 40 msec. Scalp current density topographic mapping, source analysis, and functional magnetic resonance imaging results all localized the IC effect to bilateral LOC areas. We propose that IC sensitivity described in V2 and V1 may reflect predominantly feedback modulation from higher-tier LOC areas, where IC sensitivity first occurs. Two additional observations further support this proposal. The latency of the IC effect shifted dramatically later (approximately 120 msec) when stimuli were laterally presented, indicating that retinotopic position alters IC processing. Immediately preceding the IC effect, the VEP modulated with inducer eccentricity--the configuration effect. We interpret this to represent contributions from global stimulus parameters to scene analysis. In contrast to the IC effect, the topography of the configuration effect was restricted to central parieto-occipital scalp. PMID- 12077202 TI - Brain regions controlling nonsynergistic versus synergistic movement of the digits: a functional magnetic resonance imaging study. AB - Human hand dexterity depends on the ability to move digits independently and to combine these movements in various coordinative patterns. It is well established that the primary motor cortex (M1) is important for skillful digit actions but less is known about the role played by the nonprimary motor centers. Here we use functional magnetic resonance imaging to examine the hypothesis that nonprimary motor areas and the posterior parietal cortex are strongly activated when healthy humans move the right digits in a skillful coordination pattern involving relatively independent digit movements. A task in which flexion of the thumb is accompanied by extension of the fingers and vice versa, i.e., a learned "nonsynergistic" coordination pattern, is contrasted with a task in which all digits flex and extend simultaneously in an innate synergistic coordination pattern (opening and closing the fist). The motor output is the same in the two conditions. Thus, the difference when contrasting the nonsynergistic and synergistic tasks represents the requirement to fractionate the movements of the thumb and fingers and to combine these movements in a learned coordinative pattern. The supplementary (and cingulate) motor area, the bilateral dorsal premotor area, the bilateral lateral cerebellum, the bilateral cortices of the postcentral sulcus, and the left intraparietal cortex showed stronger activity when the subjects made the nonsynergistic flexion-extension movements of the digits than when the synergistic movements were made. These results suggest that the human neural substrate for skillful digit movement includes a sensorimotor network of nonprimary frontoparietal areas and the cerebellum that, in conjunction with M1, control the movements of the digits. PMID- 12077203 TI - Visual and anticipatory bias in three cortical eye fields of the monkey during an adaptive decision-making task. AB - To examine the role of three cortical eye fields during internally guided decision-making processes, we recorded neuronal activities in the frontal eye field (FEF), supplementary eye field (SEF), and lateral intraparietal cortex (LIP) using a free-choice delayed saccade task with two synchronized targets. Although the monkeys must perform the task in a time-locked manner, they were free to choose either the receptive field (RF) target or the nonreceptive field (nRF) target to receive reward. In all three areas we found neurons with stronger activation during trials when the monkey was going to make a saccade to the RF target (RF trials) than to the nRF target (nRF trials). Modulation occurred not only during target presentation (visual bias) but also before target presentation (anticipatory bias). The visual bias was evident as an attenuated visual response to the RF stimulus in nRF trials. The anticipatory bias, however, was seen as an enhancement of pretarget activity in the RF trials. We analyzed the activity during the 500 msec before target presentation and found that 22.5% of FEF and 31.3% of LIP neurons and 49.1% of SEF neurons showed higher activity during the RF trials. To more accurately determine when each neuron started to show preferential activity, we used a new inverse interspike interval analysis procedure. Our results suggest that although all three cortical eye fields reflect attentional and intentional aspects of sensorimotor processing, SEF plays an earlier and perhaps more cognitive role in internally guided decision-making processes for saccades. PMID- 12077204 TI - Involvement of the arcuate nucleus of the hypothalamus in interleukin-1-induced anorexia. AB - Cytokine-mediated anorexia is a component of "sickness behavior" and presents a significant obstacle in the treatment of chronic illnesses. We hypothesized an involvement of the hypothalamic arcuate nucleus (ARH) in mediating the anorexic effects of a systemic interleukin-1 (IL-1) challenge based on its content of peptidergic neurons involved in feeding, its expression of IL-1 receptors and its sensitivity to systemic IL-1. IL-1 (6 microg/kg, i.v.) was found to induce Fos expression in both pro-opiomelanocortin- and neuropeptide Y-expressing neurons in and around the ARH. Contrary to expectations, rats that had sustained lesions of the arcuate nucleus, produced by neonatal monosodium glutamate treatment, displayed a more pronounced suppression (by 25%) of food intake than nonlesioned controls when treated with IL-1 after a 20 hr fast. To confirm and further characterize this unexpected result, a second ablation method was used in a similar paradigm. Animals bearing knife cuts designed to sever major ARH projections displayed an even more accentuated loss of appetite (by 60%, relative to controls) in response to systemic IL-1. This effect exhibited at least some degree of specificity, because the knife cuts did not alter either IL-1 effects on another centrally mediated acute phase response (fever) or the anorexia produced by an alternate agent, fenfluramine. These results fail to support the hypothesized ARH mediation of IL-1-induced anorexia and may suggest rather that the net output of this cell group may serve normally to restrain cytokine-induced reductions in food intake. PMID- 12077205 TI - mu-opioid receptor-mediated antinociceptive responses differ in men and women. AB - Sex differences in the experience of clinical and experimental pain have been reported. However, the neurobiological sources underlying the variability in pain responses between sexes have not been adequately explored, especially in humans. The endogenous opioid neurotransmitters and mu-opioid receptors are centrally implicated in responses to stress, in the suppression of pain, and in the action of opiate analgesic drugs. Here we examined sex differences in the activation of the mu-opioid system in response to an intensity-controlled sustained deep-tissue pain challenge with positron emission tomography and a mu-opioid receptor selective radiotracer. Twenty-eight young healthy volunteers (14 men and 14 women) were studied during saline control and pain conditions using a double blind, randomized, and counterbalanced design. Women were scanned during the early follicular phase of their menstrual cycles after ovulatory cycles. Significant sex differences in the regional activation of the mu-opioid system in response to sustained pain were detected compared with saline controls. Men demonstrated larger magnitudes of mu-opioid system activation than women in the anterior thalamus, ventral basal ganglia, and amygdala. Conversely, women demonstrated reductions in the basal state of activation of the mu-opioid system during pain in the nucleus accumbens, an area previously associated with hyperalgesic responses to the blockade of opioid receptors in experimental animals. These data demonstrate that at matched levels of pain intensity, men and women during their follicular phase differ in the magnitude and direction of response of the mu-opioid system in distinct brain nuclei. PMID- 12077206 TI - Postural modifications and neuronal excitability changes induced by a short-term serotonin depletion during neonatal development in the rat. AB - Serotonin (5-HT) plays an important role both in the development and in the recovery of locomotion after spinalization in vertebrates. We investigated the contribution of the serotonergic system to the maturation of the lumbar motoneurons and networks in the neonatal rat. A 5-HT synthesis inhibitor, p chlorophenylalanine (PCPA), was administered daily from the first postnatal day (P0) onward. This protocol depleted serotonin in the spinal cord within 3-4 d, as demonstrated by immunohistochemistry. PCPA-treated rats exhibited postural changes characterized by lesser flexion at the knee and ankle levels and lesser extension of the hip. Posture was asymmetric, suggesting possible deficits in the interlimb coordination. Intracellular recordings were made at P3-5 from motoneurons innervating different hindlimb muscles, using the in vitro brainstem spinal cord-nerve-attached preparation. In PCPA-treated rats, the conduction velocity of motoneurons was increased, and their excitability was decreased (because of higher rehobase and input conductance) compared with sham animals. In accordance with postural observations, changes were more pronounced in hip extensor/knee flexor than in ankle extensor motoneurons. The maturation of repetitive firing properties was stopped by PCPA treatment, although PCPA, applied in vitro, had no effect on membrane properties. The spontaneous endogenously generated activity, which is a characteristic of immature networks, was increased in PCPA-treated rats, suggesting that developing lumbar networks are sensitive to 5-HT levels. Serotonin may play a critical role during development in regulating the balance between the excitability of motoneurons and that of interneurons. Interneuronal excitability is crucial for the activity dependent development of spinal cord networks. PMID- 12077207 TI - How noise contributes to contrast invariance of orientation tuning in cat visual cortex. AB - The width of the orientation tuning curves of the spike response of neurons in V1 is invariant to contrast. This property constrains the possible mechanisms underlying orientation selectivity. It has been suggested that noise circumvents the iceberg effect that would prevent contrast invariance in the purely feedforward mechanism. Here we investigate systematically how noise contributes to the contrast invariance of orientation tuning curves in V1. We study three models of increasing complexity: a simple threshold-linear firing rate model, a leaky integrate-and-fire model, and a conductance-based model. We show that the noise transmutes the threshold nonlinearity of the input-output relationships into an approximate power law without a threshold within some firing rate range. This implies that, under certain conditions which are derived here, the tuning of the neuron output is approximately contrast invariant. In particular we show that this mechanism for contrast invariance requires that the neuron firing rate must not be too large and that increasing or lowering the contrast too much destroys this invariance. We also show that if this mechanism operates in V1, the spike response, R, and average voltage response V of the neurons in V1 should vary with the contrast, C, according to R(C)gamma proportional to V(C)gamma. The exponent gamma can be estimated from the amount by which the spike tuning curve is sharpened with respect to the voltage tuning curves of the neurons. This prediction does not depend on the specifics of the model and can be tested experimentally. PMID- 12077208 TI - Time-dependent descending facilitation from the rostral ventromedial medulla maintains, but does not initiate, neuropathic pain. AB - Although injury-induced afferent discharge declines significantly over time, experimental neuropathic pain persists unchanged for long periods. These observations suggest that processes that initiate experimental neuropathic pain may differ from those that maintain such pain. Here, the role of descending facilitation arising from developing plasticity in the rostral ventromedial medulla (RVM) in the initiation and maintenance of experimental neuropathic pain was explored. Tactile and thermal hypersensitivity were induced in rats by spinal nerve ligation (SNL). RVM lidocaine blocked SNL-induced tactile and thermal hypersensitivity on post-SNL days 6-12 but not on post-SNL day 3. Lesion of RVM cells expressing mu-opioid receptors with dermorphin-saporin did not prevent the onset of SNL-induced tactile and thermal hypersensitivity, but these signs reversed to baseline levels beginning on post-SNL day 4. Similarly, lesions of the dorsolateral funiculus (DLF) did not prevent the onset of SNL-induced tactile and thermal hypersensitivity, but these signs reversed to baseline levels beginning on post-SNL day 4. Lesions of the DLF also blocked the SNL-induced increase in spinal dynorphin content, which has been suggested to promote neuropathic pain. These data distinguish mechanisms that initiate the neuropathic state as independent of descending supraspinal influences and additional mechanism(s) that require supraspinal facilitation to maintain such pain. In addition, the data indicate that these time-dependent descending influences can underlie some of the SNL-induced plasticity at the spinal level. Such time dependent descending influences driving associated spinal changes, such as the upregulation of dynorphin, are key elements in the maintenance, but not initiation, of neuropathic states. PMID- 12077209 TI - High-frequency stimulation of the subthalamic nucleus selectively reverses dopamine denervation-induced cellular defects in the output structures of the basal ganglia in the rat. AB - High-frequency stimulation (HFS) of the subthalamic nucleus (STN) is now recognized as an effective treatment for advanced Parkinson's disease, but the molecular basis of its effects remains unknown. This study examined the effects of unilateral STN HFS (2 hr of continuous stimulation) in intact and hemiparkinsonian awake rats on STN neuron metabolic activity and on neurotransmitter-related gene expression in the basal ganglia, by means of in situ hybridization histochemistry and immunocytochemistry. In both intact and hemiparkinsonian rats, this stimulation was found to induce c-fos protein expression but to decrease cytochrome oxidase subunit I mRNA levels in STN neurons. STN HFS did not affect the dopamine lesion-mediated overexpression of enkephalin mRNA or the decrease in substance P in the ipsilateral striatum. The lesion-induced increases in intraneuronal glutamate decarboxylase 67 kDa isoform (GAD67) mRNA levels on the lesion side were reversed by STN HFS in the substantia nigra, partially antagonized in the entopeduncular nucleus but unaffected in the globus pallidus. The stimulation did not affect neuropeptide or GAD67 mRNA levels in the side contralateral to the dopamine lesion or in intact animals. These data furnish the first evidence that STN HFS decreases the metabolic activity of STN neurons and antagonizes dopamine lesion-mediated cellular defects in the basal ganglia output structures. They provide molecular substrate to the therapeutic effects of this stimulation consistent with the current hypothesis that HFS blocks STN neuron activity. However, the differential impact of STN HFS on the effects of dopamine lesion among structures receiving direct STN inputs suggests that this stimulation may not cause simply interruption of STN outflow. PMID- 12077210 TI - Parallel motion processing for the initiation of short-latency ocular following in humans. AB - With the scleral search coil technique, we recorded ocular following responses elicited by either grating or plaid pattern motions. Grating motion elicited tracking responses at short latencies ( approximately 85 msec). Type I plaid motion made by summing two orthogonal moving gratings elicited ocular following with identical short latencies. Trial-by-trial vector decomposition showed that plaid-driven responses were best predicted by a vector average of the component driven responses. Similar results were found with micropatterns made of 16 Gabor patches with drifting carriers of two different orientations. "Unikinetic" plaids were constructed by summing a moving and stationary grating, with a 45 degrees orientation difference, so that component and pattern motion directions were separated by 45 degrees. Eye movements exhibited two components. Ocular following was first initiated in the grating motion direction, at ultra-short latency. A second component was initiated approximately 20 msec later, curving the responses toward the pattern motion direction. The later component was specifically, and independently, affected by both relative spatial frequency and contrast between component gratings. The early response components showed a much steeper contrast response function than the late component. These results suggest that initial ocular following is underpinned by parallel processing of component- and pattern related velocities followed by an integrative stage that computes the two dimensional surface motion. PMID- 12077211 TI - Short-term plasticity shapes the response to simulated normal and parkinsonian input patterns in the globus pallidus. AB - Basal ganglia structures show strong activity modulation during movement and synchronous bursting in Parkinson's disease. Recent work has shown that short term synaptic plasticity (STP) can play an important role in the effect of temporal activity patterns on postsynaptic targets. To determine the role of STP in the subthalamic nucleus (STN) to globus pallidus (GP) connection, which has been suggested to underlie rhythmical bursting in Parkinson's disease, we first measured STP using trains of electrical input stimulation in vitro. We found that STN inputs to GP typically show both facilitation and depression with input frequencies of 10-100 Hz and that facilitation is dominant for the first few inputs in a train but that depression takes over subsequently. We quantified the strength and time course of facilitation and depression using a computational model of STP. Using the STP model, we constructed synaptic conductance patterns of normal and Parkinsonian STN activity and applied these conductances to GP neurons in vitro using the technique of dynamic clamping. We show that STP controls the slope and shape of the function describing the steady-state level of GP neuron firing in response to different levels of STN input. In addition, we show that STP modulates responses of GP neurons to bursts and pauses in the input pattern. These findings indicate that STP plays an important role in modulating both spike rates and temporal patterns of GP activity in the normal state, as well as in Parkinson's disease. PMID- 12077212 TI - Activation of ventral tegmental area cells by the bed nucleus of the stria terminalis: a novel excitatory amino acid input to midbrain dopamine neurons. AB - We examined the role of excitatory amino acids (EAAs) in the activation of midbrain dopaminergic (DA) neurons evoked by stimulation of the ventromedial and ventrolateral (subcommissural) bed nucleus of the stria terminalis (vBNST). Using anesthetized rats and extracellular recording techniques, we found that 84.8% of ventral tegmental area (VTA) DA neurons were activated synaptically by single pulse electrical stimulation of the vBNST. In contrast, similar stimulation did not affect the activity of presumed GABA neurons in the VTA. Three characteristic responses were observed in VTA DA neurons: short latency activation (<25 msec; 55.1% of cells), long latency activation (>65 msec; 56% of cells), and inhibition (61.8% of cells, usually followed by long latency excitation). Microinfusion of antagonists of EAA receptors (3 mm kynurenic acid, 100 microm AP-5, or 50 microm CNQX) from a micropipette adjacent to the recording electrode significantly reduced both short and long latency activations evoked in DA neurons by vBNST stimulation. Specific responses were attenuated similarly by AP-5 alone, CNQX alone, or a cocktail of AP-5+CNQX, indicating that joint activation of NMDA plus non-NMDA receptors was required. Stimulation of the vBNST by local microinfusion of glutamate increased the firing and bursting activity of VTA DA neurons. Similar microinfusion of GABA decreased bursting of VTA DA neurons without altering their firing rate. Retrograde and anterograde labeling and antidromic activation of vBNST neurons by VTA stimulation confirmed a direct projection from the vBNST to the VTA. These results reveal that inputs from the vBNST exert a strong excitatory influence on VTA DA neurons mediated by both NMDA and non-NMDA receptors. PMID- 12077213 TI - Phosphodiesterase 1B knock-out mice exhibit exaggerated locomotor hyperactivity and DARPP-32 phosphorylation in response to dopamine agonists and display impaired spatial learning. AB - Using homologous recombination, we generated mice lacking phosphodiesterase mediated (PDE1B) cyclic nucleotide-hydrolyzing activity. PDE1B(-/-) mice showed exaggerated hyperactivity after acute D-methamphetamine administration. Striatal slices from PDE1B(-/-) mice exhibited increased levels of phospho-Thr34 DARPP-32 and phospho-Ser845 GluR1 after dopamine D1 receptor agonist or forskolin stimulation. PDE1B(-/-) and PDE1B(+/-) mice demonstrated Morris maze spatial learning deficits. These results indicate that enhancement of cyclic nucleotide signaling by inactivation of PDE1B-mediated cyclic nucleotide hydrolysis plays a significant role in dopaminergic function through the DARPP-32 and related transduction pathways. PMID- 12077214 TI - Cognitive slowing in Parkinson's disease: a behavioral evaluation independent of motor slowing. AB - Parkinson's disease (PD) is attributable primarily to depletion of dopamine in the basal ganglia, but the full effects of this depletion are unknown. It is well known that PD involves motor slowing, and although it is not easy to distinguish between the motor and cognitive components of behavior, clinical observations suggest that cognitive processing may also be compromised. However, it remains unclear whether such cognitive involvement exists, and if so, to what extent. Previous studies of cognitive slowing in PD have yielded conflicting results. This may be attributable to variations in experimental procedures, because most of the experiments used reaction-time tasks, which are inevitably confounded by motor components. In the present study, we evaluated the speed of cognitive processing in patients with PD without bradykinesia as a variable. We developed a mental-operation task that required serial updating of mental representations in response to a series of visual stimuli. By changing the speed of visual presentation and evaluating performance accuracy, the speed of cognitive processing was assessed independently of motor slowing. Cognitive impairment in PD became evident when higher speeds of cognitive processing (verbal more so than spatial) were required. In addition, cognitive slowing and motor slowing were significantly correlated. The results of the present study suggest that slowing in PD is not restricted to the motor domain but can be generally observed in other domains of behavior, including cognitive mental operations. PMID- 12077215 TI - Androgens protect against apolipoprotein E4-induced cognitive deficits. AB - Compared with apolipoprotein (apo) E2 and E3, apoE4 increases the risk of Alzheimer's disease (AD), but it remains unknown how apoE4 affects neuronal function. ApoE4 interacts with female gender, further increasing the risk of AD and decreasing treatment response. Female mice are also more susceptible to apoE4 induced impairments of spatial learning and memory than male mice. To assess the role of sex steroids in this process, we studied mice deficient in mouse apoE (Apoe(-/-)) and expressing human apoE4 or apoE3 in the brain at comparable levels. Even brief periods of androgen treatment improved the memory deficits of female apoE4 mice. Female apoE3 mice had no memory deficits and did not benefit from the treatment. ApoE4 male mice, which performed normally in a water-maze test at baseline, developed prominent deficits in spatial learning and memory after blockade of androgen receptors (ARs), whereas apoE3 male mice did not. Untreated apoE4 mice had significantly lower cytosolic AR levels in the neocortex than wild-type, Apoe(-/-), and apoE3 mice. Improved memory in androgen-treated female apoE4 mice was associated with increased cytosolic AR levels. Our findings suggest that apoE4 contributes to cognitive decline by reducing AR levels in the brain, and that stimulating AR-dependent pathways can reverse apoE4-induced cognitive deficits. PMID- 12077216 TI - Dopamine modulates excitability of spiny neurons in the avian basal ganglia. AB - The neural substrate of vocal learning in songbirds is an accessible system for studying motor learning and motor control in vertebrates. In the so-called song system, the anterior forebrain pathway (AFP), which is essential for song learning, resembles the mammalian basal ganglia-thalamocortical loop in its macroscopic organization, neuronal intrinsic properties, and microcircuitry. Area X, the first station in the AFP, and the surrounding lobus parolfactorius (LPO), are both parts of the avian basal ganglia. Like their mammalian counterparts, they receive dense dopaminergic innervation from the midbrain, but the physiological functions of this projection remain unclear. In this study, we investigated the effect of dopamine (DA) on excitability of spiny neurons in area X and LPO. We recorded from neurons in brain slices of adult zebra finches and Bengalese finches, using whole-cell and perforated-patch recording techniques in current-clamp configuration. We found that DA modulates excitability in spiny neurons; activation of D1- and D2-like DA receptors enhances and reduces excitability, respectively. These effects are similar to those observed in the mammalian neostriatum, with the main difference being that D1-like DA receptor activation enhances excitability in avian spiny neurons at hyperpolarized states. Our findings also indicate that some spiny neurons express both receptor types and suggest that receptor colocalization in the entire population can account for the spectrum of DA actions. The diversity of DA actions enables the DA system to fine-tune the dynamics of the song system and allows flexible control over song learning and production. PMID- 12077217 TI - The group I metabotropic glutamate receptor mGluR5 is required for fear memory formation and long-term potentiation in the lateral amygdala. AB - The group I metabotropic glutamate receptor subtype mGluR5 has been shown to play a key role in the modulation of synaptic plasticity. The present experiments examined the function of mGluR5 in the circuitry underlying Pavlovian fear conditioning using neuroanatomical, electrophysiological, and behavioral techniques. First, we show using immunocytochemical and tract-tracing methods that mGluR5 is localized to dendritic shafts and spines in the lateral nucleus of the amygdala (LA) and is postsynaptic to auditory thalamic inputs. In electrophysiological experiments, we show that long-term potentiation at thalamic input synapses to the LA is impaired by bath application of a specific mGluR5 antagonist, 2-methyl-6-(phenyle-thynyl)-pyridine (MPEP), in vitro. Finally, we show that intra-amygdala administration of MPEP dose-dependently impairs the acquisition, but not expression or consolidation, of auditory and contextual fear conditioning. Collectively, the results of this study indicate that mGluR5 in the LA plays a crucial role in fear conditioning and in plasticity at synapses involved in fear conditioning. PMID- 12077218 TI - Fos imaging reveals that lesions of the anterior thalamic nuclei produce widespread limbic hypoactivity in rats. AB - Activity of the immediate early gene c-fos was compared in rats with neurotoxic lesions of the anterior thalamic nuclei and in surgical controls. Fos levels were measured after rats had been placed in a novel room and allowed to run up and down preselected arms of a radial maze. An additional control group showed that in normal rats, this exposure to a novel room leads to a Fos increase in a number of structures, including the anterior thalamic nuclei and hippocampus. In contrast, rats with anterior thalamic lesions were found to have significantly less Fos-positive cells in an array of sites, including the hippocampus (dorsal and ventral), retrosplenial cortex, anterior cingulate cortex, and prelimbic cortex. These results show that anterior thalamic lesions disrupt multiple limbic brain regions, producing hypoactivity in sites associated in rats with spatial memory. Because many of the same sites are implicated in memory processes in humans (e.g., the hippocampus and retrosplenial cortex), this hypoactivity might contribute to diencephalic amnesia. PMID- 12077219 TI - NMDA receptors and L-type voltage-gated calcium channels contribute to long-term potentiation and different components of fear memory formation in the lateral amygdala. AB - Long-term potentiation (LTP) at sensory input synapses to the lateral amygdala (LA) is a candidate mechanism for memory storage during fear conditioning. We evaluated the effect of L-type voltage-gated calcium channel (VGCC) and NMDA receptor (NMDAR) blockade in LA on LTP at thalamic input synapses induced by two different protocols in vitro and on fear memory in vivo. When induced in vitro by pairing weak presynaptic stimulation with strong (spike eliciting) postsynaptic depolarization, LTP was dependent on VGCCs and not on NMDARs, but, when induced by a form of tetanic stimulation that produced prolonged postsynaptic depolarization (but not spikes), LTP was dependent on NMDARs and not on VGCCs. In behavioral studies, bilateral infusions of NMDAR antagonists into the LA impaired both short-term and long-term memory of fear conditioning, whereas VGCC blockade selectively impaired long-term memory formation. Collectively, the results suggest that two pharmacologically distinct forms of LTP can be isolated in the LA in vitro and that a combination of both contribute to the formation of fear memories in vivo at the cellular level. PMID- 12077220 TI - Cutting edge: chemical dominance does not relate to immunodominance: studies of the CD4+ T cell response to a model antigen. AB - We have quantitated the major families of peptides from hen egg lysozyme (HEL) presented by MHC class II I-A(k) molecules. One striking feature is that the four epitopes are presented at levels that differ by as much as 200- to 300-fold. In these studies, we describe the CD4(+) T cell response to each epitope after immunization with several doses of hen egg lysozyme protein. Although fewer T cells were generated at lower doses, the surprising finding was the responses to all four peptides were maintained. The relative number of T cell clones to each of the four epitopes was influenced to a very limited degree by their levels of presentation at the lowest dose. In conclusion, under strong stimulatory conditions, there is not a direct relationship between levels of peptide presentation and the T cell responses. PMID- 12077222 TI - Cutting edge: role of Toll-like receptor 1 in mediating immune response to microbial lipoproteins. AB - The Toll-like receptor (TLR) family acts as pattern recognition receptors for pathogen-specific molecular patterns (PAMPs). TLR2 is essential for the signaling of a variety of PAMPs, including bacterial lipoprotein/lipopeptides, peptidoglycan, and GPI anchors. TLR6 associates with TLR2 and recognizes diacylated mycoplasmal lipopeptide along with TLR2. We report here that TLR1 associates with TLR2 and recognizes the native mycobacterial 19-kDa lipoprotein along with TLR2. Macrophages from TLR1-deficient (TLR1(-/-)) mice showed impaired proinflammatory cytokine production in response to the 19-kDa lipoprotein and a synthetic triacylated lipopeptide. In contrast, TLR1(-/-) cells responded normally to diacylated lipopeptide. TLR1 interacts with TLR2 and coexpression of TLR1 and TLR2 enhanced the NF-kappaB activation in response to a synthetic lipopeptide. Furthermore, lipoprotein analogs whose acylation was modified were preferentially recognized by TLR1. Taken together, TLR1 interacts with TLR2 to recognize the lipid configuration of the native mycobacterial lipoprotein as well as several triacylated lipopeptides. PMID- 12077221 TI - Cutting edge: molecular portrait of human autoimmune disease. AB - Autoimmune diseases affect 3-5% of the population, are mediated by the immune response to self-Ags, and are characterized by the site of tissue destruction. We compared expression levels of >4,000 genes in PBMC of control individuals before and after immunization to those of individuals with four distinct autoimmune diseases. The gene expression profile of the normal immune response exhibits coordinate changes in expression of genes with related functions over time. In contrast, each individual from all autoimmune diseases displays a similar gene expression profile unrelated to the pattern of the immunized group. To our surprise, genes with a distinct expression pattern in autoimmunity are not necessarily "immune response" genes, but are genes that encode proteins involved in apoptosis, cell cycle progression, cell differentiation, and cell migration. PMID- 12077223 TI - Extracellular adenine nucleotides inhibit the activation of human CD4+ T lymphocytes. AB - ATP has been reported to inhibit or stimulate lymphoid cell proliferation, depending on the origin of the cells. Agents that increase cAMP, such as PGE(2), inhibit human CD4(+) T cell activation. We demonstrate that several ATP derivatives increase cAMP in both freshly purified and activated human peripheral blood CD4(+) T cells. The rank order of potency of the various nucleotides was: adenosine 5'-O-(3-thiotriphosphate) (ATPgammaS) approximately 2'- and 3'-O-(4 benzoylbenzoyl) ATP (BzATP) > ATP > 2-methylthio-ATP >> dATP, 2-propylthio beta,gamma-dichloromethylene-D-ATP, UDP, UTP. This effect did not involve the activation of A(2)Rs by adenosine or the synthesis of prostaglandins. ATPgammaS had no effect on cytosolic calcium, whereas BzATP induced an influx of extracellular calcium. ATPgammaS and BzATP inhibited secretion of IL-2, IL-5, IL 10, and IFN-gamma; expression of CD25; and proliferation after activation of CD4(+) T cells by immobilized anti-CD3 and soluble anti-CD28 Abs, without increasing cell death. Taken together, our results suggest that extracellular adenine nucleotides inhibit CD4(+) T cell activation via an increase in cAMP mediated by an unidentified P2YR, which might thus constitute a new therapeutic target in immunosuppressive treatments. PMID- 12077224 TI - Ly-49s3 is a promiscuous activating rat NK cell receptor for nonclassical MHC class I-encoded target ligands. AB - Previous studies of the rapid rejection of MHC-disparate lymphocytes in rats, named allogeneic lymphocyte cytotoxicity, have indicated that rat NK cells express activating receptors for nonclassical MHC class I allodeterminants from the RT1-C/E/M region. Using an expression cloning system that identifies activating receptors associated with the transmembrane adapter molecule DAP12, we have cloned a novel rat Ly-49 receptor that we have termed Ly-49 stimulatory receptor 3 (Ly-49s3). A newly generated anti-Ly-49s3 Ab, mAb DAR13, identified subpopulations of resting and IL-2-activated NK cells, but not T or B lymphocytes. Depletion of Ly-49s3-expressing NK cells drastically reduced alloreactivity in vitro, indicating that this subpopulation is responsible for a major part of the observed NK alloreactivity. DAR13-mediated blockade of Ly-49s3 inhibited killing of MHC-congenic target cells from the av1, n, lv1, and c haplotypes, but not from the u or b haplotypes. A putative ligand was mapped to the nonclassical MHC class I region (RT1-C/E/M) using intra-MHC recombinant strains. Relative numbers of Ly-49s3(+) NK cells were reduced, and surface levels of Ly-49s3 were lower, in MHC congenic strains expressing the putative Ly-49s3 ligand(s). In conclusion, we have identified a novel Ly-49 receptor that triggers rat NK cell-mediated responses. PMID- 12077225 TI - NKT cell-derived RANTES recruits APCs and CD8+ T cells to the spleen during the generation of regulatory T cells in tolerance. AB - The induction of peripheral tolerance via immune privileged sites such as the eye requires splenic colocalization of NKT cells and CD1d(+) tolerogenic F4/80(+) APCs, both of which are needed for the generation of CD8(+)-regulatory T (Tr) cells. Whereas tolerogenic APCs secrete the chemokine macrophage-inflammatory protein-2 for the purpose of recruiting NKT cells, the signals responsible for recruiting potential Tr cells and additional APCs to the spleen are not known. Here we examined the ability of CD1d-stimulated NKT cells to produce chemokines that can recruit other cells needed for tolerance. Our results show that NKT cells stimulated by either CD1d-transfected fibroblasts in vitro or CD1d(+) tolerogenic APCs both in vivo and ex vivo produced RANTES in a CD1d-dependent manner. The requirement for RANTES in tolerance was demonstrated by studies in which RANTES blockade in vivo prevented not only APC accumulation in the spleen but also the generation of CD8(+) Tr cells that suppress Th1 immunity. Thus, CD1d restricted NKT cells provide critical signals for orchestrating the accumulation of cells needed for tolerance induction. These data expand our current knowledge of RANTES beyond its role in Th1 immune responses to show its importance in tolerance induction and add a novel aspect to our understanding of the role of NKT cells in tolerance. Understanding the precise mechanisms involved in tolerance induction may lead to more effective therapeutic strategies for autoimmunity and graft rejection. PMID- 12077226 TI - Memory functions and death proneness in three CD4+CD45RO+ human T cell subsets. AB - We propose a classification of human CD4(+)CD45RO(+) memory T cells into three new subsets based on cell surface expression levels of CD43. The first subset consists of cells whose CD43 expression is relatively high; this subset also contains the highest proportion of recall Ag-reactive precursors, and its constituent cells respond far more strongly than cells in either of the other subsets to immobilized CD3 Ab in addition to secreting substantially more IFN gamma and IL-4. Cells of the second subset express similar levels of CD43 to naive cells, and they also respond weakly to TCR-mediated stimuli as judged by either their ability to proliferate or capacity for cytokine production. The third subsets consists of cells whose CD43 expression levels are clearly down regulated; its cells appear to be anergic to TCR-mediated stimuli, and when examined ex vivo many of them appear to be undergoing either spontaneous apoptosis via a caspase-independent pathway or Fas-mediated apoptosis via a caspase-dependent pathway, even in the resting state. An analysis of telomere lengths revealed that the typical telomere of a cell in the second subset was significantly longer than the typical telomere in the first or third subset. Taken together, these results appear to indicate that CD4(+)CD45RO(+) T cells fall into three functionally differing subsets, one being a subset of cells with fully matured memory phenotype, a second being a less mature subset of cells that retain longer telomeres and whose memory functionality is marginal, and a third consisting of anergic cells that give every appearance of being death-prone and/or in the process of dying. PMID- 12077227 TI - Disruption of a single Pten allele augments the chemotactic response of B lymphocytes to stromal cell-derived factor-1. AB - The tumor suppressor, Pten, has emerged as a critical negative regulator of phosphatidylinositol-3-kinase-dependent intracellular signaling pathways responsible for phenomena such as cellular adhesion, proliferation, and apoptosis. Herein, we present evidence that Pten regulates chemokine-dependent events in B lymphocytes. Primary B cells isolated from Pten(+/-) mice demonstrated increased responsiveness to stromal cell-derived factor-1-induced chemotaxis. This was accompanied by an elevated level of protein kinase B phosphorylation on Ser(473). Our results suggest not only that Pten may be an important regulator of stromal cell-derived factor-1-directed chemotaxis, but also that Pten heterozygosity is associated with increased cellular sensitivity to this chemokine, likely via dysregulation of events lying downstream of phosphatidylinositol-3-kinase. These observations suggest a mechanism by which loss of a single Pten allele may confer a selective advantage on cells during multistep tumor progression. PMID- 12077228 TI - 2B4 is constitutively associated with linker for the activation of T cells in glycolipid-enriched microdomains: properties required for 2B4 lytic function. AB - 2B4 is a receptor belonging to the Ig superfamily and is found on all murine NK cells as well as a small subset of T cells. Previous studies have found that cross-linking of the 2B4 receptor results in both increased cytotoxicity and IFN gamma secretion. We have discovered that 2B4 from transfected NK and T cell lines, as well as from primary murine cells, coimmunoprecipitates with the phosphoprotein linker for the activation of T cells (LAT), which is essential for TCR-mediated signaling. This association is independent of both 2B4 phosphorylation and the cytoplasmic tail of 2B4. We have found that, along with LAT, 2B4 is constitutively located in glycolipid-enriched microdomains of the plasma membrane. In fact, 2B4 appears to associate with LAT only when it localizes to glycolipid-enriched microdomains. This localization of 2B4 occurs due to a CxC cysteine motif found in the transmembrane region, as determined by mutagenesis studies. 2B4-mediated cytotoxicity is defective in the absence of LAT, indicating that LAT is a required intermediate for 2B4 signal transduction. However, we have also shown that LAT association alone is not sufficient for maximal 2B4 activation. PMID- 12077229 TI - IL-12 administration leads to a transient depletion of T cells, B cells, and APCs and concomitant abrogation of the HLA-A2.1-restricted CTL response in transgenic mice. AB - The injection of a mixture of bona fide T cell epitopes can lead to the occurrence of immunodominance, meaning that the immune response is focused on the recognition of a single epitope or a small portion of the epitopes injected. We have previously demonstrated that the administration of rIL-12 can counteract immunodominance in BALB/c mice. In this study, we show that the administration of rIL-12 to HLA-A2.1 transgenic mice (A2k(b) mice) abrogates specifically the immune response against HLA-A2.1-restricted HIV epitopes in the spleen. This lack of immune response is most probably due to a transient depletion of B cells, T cells, macrophages, and dendritic cells in this organ. Therefore, our study explains the mechanism of immunosuppression by rIL-12 in vivo. PMID- 12077230 TI - Src-dependent Syk activation controls CD69-mediated signaling and function on human NK cells. AB - CD69 C-type lectin receptor represents a functional triggering molecule on activated NK cells, capable of directing their natural killing function. The receptor-proximal signaling pathways activated by CD69 cross-linking and involved in CD69-mediated cytotoxic activity are still poorly understood. Here we show that CD69 engagement leads to the rapid and selective activation of the tyrosine kinase Syk, but not of the closely related member of the same family, ZAP70, in IL-2-activated human NK cells. Our results indicate the requirement for Src family kinases in the CD69-triggered activation of Syk and suggest a role for Lck in this event. We also demonstrate that Syk and Src family tyrosine kinases control the CD69-triggered tyrosine phosphorylation and activation of phospholipase Cgamma2 and the Rho family-specific exchange factor Vav1 and are responsible for CD69-triggered cytotoxicity of activated NK cells. The same CD69 activated signaling pathways are also observed in an RBL transfectant clone, constitutively expressing the receptor. These data demonstrate for the first time that the CD69 receptor functionally couples to the activation of Src family tyrosine kinases, which, by inducing Syk activation, initiate downstream signaling pathways and regulate CD69-triggered functions on human NK cells. PMID- 12077231 TI - CD123bright plasmacytoid predendritic cells: progenitors undergoing cell fate conversion? AB - CD123(bright) plasmacytoid cells (PC) and CD1c(+) peripheral blood myeloid dendritic cells (DC) are two human DC precursors that can be expanded in vivo by Fms-like tyrosine kinase 3 ligand (FL). It has been proposed that PC and myeloid CD1c(+) DC may represent two distinct lineages of DC. However, the phylogenetic affiliation of PC and its relationship with myeloid DC remain controversial. Here we show that CD123(bright)HLA-DR(+) PC from FL-treated healthy volunteers can be divided into mutually exclusive subsets that harbor either lymphoid or myeloid features. Lymphoid-like PC represent the majority of PC and include pTalpha-, CD3epsilon-, and CD7-expressing cells. They exhibit TCR-beta gene loci in germline configuration and show low allostimulatory capacity, but produce type I IFN upon virus infection and can be differentiated in vitro into potent APC. Myeloid-like PC represent a minor fraction of the total PC population. They exhibit a striking PC/myeloid DC intermediate phenotype (CD5(+)CD11c(low)CD45RA(low)CD45RO(-)CD101(+)), produce proinflammatory cytokines, and do not require in vitro maturation to act as potent APCs. We propose that, rather than forming a lineage, PC might represent a population of lymphoid cells undergoing an in vivo cell fate conversion from a lymphoid to a myeloid cell type. PMID- 12077232 TI - Using tolerance induced via the anterior chamber of the eye to inhibit Th2 dependent pulmonary pathology. AB - Anterior chamber-associated immune deviation (ACAID), a manifestation of ocular immune privilege, prevents Th1-dependent delayed hypersensitivity from developing in response to eye-derived Ags, thereby preserving vision. Since Th2-type cells have recently been shown to mediate destructive inflammation of the cornea, we wondered whether pre-emptive induction of ACAID could inhibit Th2 responses. Using a murine model of OVA -specific, Th2-dependent pulmonary inflammation, we pretreated susceptible mice by injecting OVA alone into the anterior chamber, or by injecting OVA-pulsed, TGF-beta2-treated peritoneal exudate cells i.v. These mice were then immunized with OVA plus alum strategy that generates Th2-mediated OVA-specific pulmonary pathology. When pretreated mice were challenged intratracheally with OVA, their bronchoalveolar lavage fluids contained far fewer eosinophils and significantly less IL-4, IL-5, and IL-13 compared with that of positive, nonpretreated controls. Similarly, lung-draining lymph node cells of pretreated mice secreted significantly less IL-4, IL-5, and IL-13 when challenged in vitro with OVA. Moreover, sera from pretreated mice contained much lower titers of OVA-specific IgE Abs. We conclude that Ags injected into the anterior chamber of the eye impair both Th1 and Th2 responses. These results reduce the likelihood that ACAID regulates Th1 responses via a Th2-like mechanism. Thus, immune privilege of the eye regulates inflammation secondary to both Th1- and Th2 type immune responses. PMID- 12077233 TI - Dynamics of memory T cell proliferation under conditions of heterologous immunity and bystander stimulation. AB - By examining adoptively transferred CSFE-labeled lymphocytic choriomeningitis virus (LCMV)-immune donor T cells in Thy-1 congenic hosts inoculated with viruses or with the cytokine inducer poly(I:C), strikingly different responses of bona fide memory T cells were found in response to different stimuli. Poly(I:C) (cytokine) stimulation caused a limited synchronized division of memory CD8 T cells specific to each of five LCMV epitopes, with no increase and sometimes a loss in number, and no change in their epitope hierarchy. Homologous LCMV infection caused more than seven divisions of T cells specific for each epitope, with dramatic increases in number and minor changes in hierarchy. Infections with the heterologous viruses Pichinde and vaccinia (VV) caused more than seven divisions and increases in number of T cells specific to some putatively cross reactive but not other epitopes and resulted in substantial changes in the hierarchy of the LCMV-specific T cells. Hence, there can be memory T cell division without proliferation (i.e., increase in cell number) in the absence of Ag and division with proliferation in the presence of Ag from homologous or heterologous viruses. Heterologous protective immunity between viruses is not necessarily reciprocal, given that LCMV protects against VV but VV does not protect against LCMV. VV elicited proliferation of LCMV-induced CD8 and CD4 T cells, whereas LCMV did not elicit proliferation of VV-induced T cells. Thus, depending on the pathogen and the sequence of infection, a heterologous agent may selectively stimulate the memory pool in patterns consistent with heterologous immunity. PMID- 12077234 TI - Exogenous peptides delivered by ricin require processing by signal peptidase for transporter associated with antigen processing-independent MHC class I-restricted presentation. AB - In this study we demonstrate that a disarmed version of the cytotoxin ricin can deliver exogenous CD8(+) T cell epitopes into the MHC class I-restricted pathway by a TAP-independent, signal peptidase-dependent pathway. Defined viral peptide epitopes genetically fused to the N terminus of an attenuated ricin A subunit (RTA) that was reassociated with its partner B subunit were able to reach the early secretory pathway of sensitive cells, including TAP-deficient cells. Successful processing and presentation by MHC class I proteins was not dependent on proteasome activity or on recycling of MHC class I proteins, but rather on a functional secretory pathway. Our results demonstrated a role for signal peptidase in the generation of peptide epitopes associated at the amino terminus of RTA. We showed, first, that potential signal peptide cleavage sites located toward the N terminus of RTA can be posttranslationally cleaved by signal peptidase and, second, that mutation of one of these sites led to a loss of peptide presentation. These results identify a novel MHC class I presentation pathway that exploits the ability of toxins to reach the lumen of the endoplasmic reticulum by retrograde transport, and suggest a role for endoplasmic reticulum signal peptidase in the processing and presentation of MHC class I peptides. Because TAP-negative cells can be sensitized for CTL killing following retrograde transport of toxin-linked peptides, application of these results has direct implications for the development of novel vaccination strategies. PMID- 12077235 TI - Antigen presentation capacity and cytokine production by murine splenic dendritic cell subsets upon Salmonella encounter. AB - Salmonella typhimurium is an intracellular bacterium that replicates in the spleen and mesenteric lymph nodes (MLN) of orally infected mice. However, little is known about the Ag presentation and cytokine production capacity of dendritic cells (DC), particularly CD8alpha(+), CD8alpha(-)CD4(-), and CD8alpha(-)CD4(+) DC, from these organs in response to Salmonella. Infection of purified splenic DC with S. typhimiurium expressing green fluorescent protein (GFP) and OVA revealed that all three splenic DC subsets internalize bacteria, and splenic as well as MLN DC process Salmonella for peptide presentation. Furthermore, presentation of Salmonella Ags on MHC-I and MHC-II was evident in both CD8alpha(+) and CD8alpha( ) splenic DC subsets. Direct ex vivo analysis of splenic DC from mice infected with GFP-expressing Salmonella showed that all three subsets harbored bacteria, and splenic DC purified from mice given Salmonella-expressing OVA presented OVA derived peptides on MHC-I and MHC-II. Cytokine production analyzed by intracellular staining of splenic DC infected with GFP-expressing Salmonella revealed that TNF-alpha was produced by a large percentage of CD8alpha(-) DC, while only a minor proportion of CD8alpha(+) DC produced this cytokine following bacterial exposure. In contrast, the greatest number of IL-12p40-producing DC were among CD8alpha(+) DC. Experiments inhibiting bacterial uptake by cytochalasin D as well as use of a Transwell system revealed that bacterial contact, but not internalization, was required for cytokine production. Thus, DC in sites of Salmonella replication and T cell activation, spleen and MLN, respond to bacterial encounter by Ag presentation and produce cytokines in a subset specific fashion. PMID- 12077236 TI - Pertussis toxin modulates the immune response to neuroantigens injected in incomplete Freund's adjuvant: induction of Th1 cells and experimental autoimmune encephalomyelitis in the presence of high frequencies of Th2 cells. AB - Pertussis toxin (PT) has been widely used to facilitate the induction of experimental autoimmune encephalomyelitis (EAE) in rodents. It has been suggested that this microbial product promotes EAE by opening up the blood-brain barrier and thereby facilitates the migration of pathogenic T cells to the CNS. However, PT has other biological effects that could contribute to its activity in EAE, such as enhancing the cytokine production by T cells and induction of lymphocytosis. In this work, we investigated the effects of PT on the pathogenicity, cytokine differentiation, and clonal sizes of neuroantigen reactive T cells in EAE in mice. Our results show that PT prevented the protection from EAE conferred by injection of PLPp139-151 in IFA and induced high frequencies of peptide-specific Th1 cells and disease. Interestingly, the mice developed EAE despite the simultaneous vigorous clonal expansion of PLPp139-151 specific Th2 cells. The data indicate that the Th2 cells in this model neither were protective against EAE nor promoted the disease. Furthermore, the results suggested that the effects of the toxin on neuroantigen-reactive T cells were promoted by the PT-induced activation of APCs in lymphoid tissues and the CNS. Together, the results suggest that microbial products, such as PT, could contribute to the initiation of autoimmune disease by modulating the interaction between the innate and adaptive immune system in the response to self Ags. PMID- 12077237 TI - A ligand for the murine NK activation receptor Ly-49D: activation of tolerized NK cells from beta 2-microglobulin-deficient mice. AB - Mouse NK cells express inhibitory NK receptors that recognize target cell MHC class I molecules and activation receptors that are less well defined. The Ly-49D activation receptor on C57BL/6 NK cells recognizes Chinese hamster ovary cells and triggers natural killing. In this study, we demonstrate that a Chinese hamster classical MHC class I molecule is the ligand for Ly-49D in a reporter gene assay system as well as in NK cell killing assays. Ly-49D recognizes the Chinese hamster class I molecule better when it is expressed with Chinese hamster beta(2)-microglobulin (beta(2)m) than murine beta(2)m. However, it is still controversial that Ly-49D recognizes H-2D(d), as we were unable to demonstrate the specificity previously reported. Using this one ligand-one receptor recognition system, function of an NK activation receptor was, for the first time, investigated in NK cells that are tolerized in beta(2)m-deficient mice. Surprisingly, Ly-49D-killing activity against ligand-expressing targets was observed with beta(2)m-deficient mouse NK cells, albeit reduced, even though "tolerized" function of Ly-49D was expected. These results indicate that Ly-49D specifically recognizes the Chinese hamster MHC class I molecule associated with Chinese hamster beta(2)m, and indicate that the Ly-49D NK cell activation receptor is not tolerized in beta(2)m deficiency. PMID- 12077238 TI - Synergistic antitumor effect of bispecific CD19 x CD3 and CD19 x CD16 diabodies in a preclinical model of non-Hodgkin's lymphoma. AB - To target NK cells against non-Hodgkin's lymphoma, we constructed a bispecific diabody (BsDb) with reactivity against both human CD19 and FcgammaRIII (CD16). Bacterially produced CD19 x CD16 BsDb specifically interacted with both CD19(+) and CD16(+) cells and exhibited significantly higher apparent affinity and slower dissociation from the tumor cells than from effector cells. It was able to induce specific lysis of tumor cells in the presence of isolated human NK cells or nonfractionated PBLs. The combination of the CD19 x CD16 BsDb with a previously described CD19 x CD3 BsDb and CD28 costimulation significantly increased the lytic potential of human PBLs. Treatment of SCID mice bearing an established Burkitt's lymphoma (5 mm in diameter) with human PBLs, CD19 x CD16 BsDb, CD19 x CD3 BsDb, and anti-CD28 mAb resulted in the complete elimination of tumors in 80% of animals. In contrast, mice receiving human PBLs in combination with either diabody alone showed only partial tumor regression. These data clearly demonstrate the synergistic effect of small recombinant bispecific molecules recruiting different populations of human effector cells to the same tumor target. PMID- 12077240 TI - Absence of the CD1 molecule up-regulates antitumor activity induced by CpG oligodeoxynucleotides in mice. AB - The role of NKT cells on antitumor activity of CpG oligodeoxynucleotides (ODNs) was evaluated by peritumoral injections of CpG-ODNs in s.c. melanoma-bearing mice of strains differing in the number of NKT cells (athymic nude mice, recombination activating gene(-/-)/transgenic V(alpha)14/Vbeta8.2 mice that generate NKT cells; J(alpha)281(-/-) mice and CD1(-/-) mice, which both have a strongly reduced number of NKT cells; and C57BL/6 wild-type mice). Tumor growth was significantly inhibited in strains enriched or depleted of NKT cells. The two murine strains having a reduced number of NKT cells differed significantly in the CpG-dependent tumor growth inhibition: in J(alpha)281(-/-) mice this inhibition was superimposable to that observed in C57BL/6 mice, while in CD1(-/-) mice the inhibition was dramatic. The increased tumor inhibition in CD1(-/-) correlated with a significantly higher ratio of IFN-gamma-IL-4 production in response to CpG as compared with C57BL/6 and J(alpha)281(-/-) mice. Experiments in which preparations of APCs and lymphocytes of the three strains were mixed showed that in the presence of APCs not expressing CD1, the production of CpG-ODN-induced type 1 cytokines was higher. Phenotype analysis of IFN-gamma- and IL-4-producing cells revealed that the differences between CD1(-/-) and C57BL/6 in the production of these two cytokines were mainly due to CD3(+) T lymphocytes. These data point to a regulatory role for the CD1 molecule in antitumor activity induced by danger signals, independently of V(alpha)14 NKT cells. The identification of a CD1-dependent suppressive subpopulation(s) might have important implications for the study of tolerance in the context of cancer, autoimmunity, and transplantation. PMID- 12077239 TI - Antigen-dependent release of IFN-gamma by cytotoxic T cells up-regulates Fas on target cells and facilitates exocytosis-independent specific target cell lysis. AB - Effector cytolytic T (Tc) lymphocytes, deficient in the exocytosis-mediated pathway of target cell lysis, induce Fas on target cells and, in turn, delayed cell death and apoptosis via the Fas ligand-Fas interaction. The induction of Fas can be blocked by anti- IFN-gamma Abs. This Fas up-regulation on initially Fas negative target cells is not mediated by TCR-MHC/peptide signaling per se, but by secreted IFN-gamma from Tc cells after Ag engagement. The Fas up-regulation by Tc cells can be mimicked by treatment of target cells with rIFN-gamma. Tc cells from IFN-gamma knockout mice do not induce Fas expression on target cells. Tc cell mediated Fas expression on third party, bystander, target cells does not enhance their susceptibility to lysis by these nominal effector cells. The results are discussed as to the possible relevance of the phenomenon in efficiency and regulation of the Tc cell response to infections by viruses. PMID- 12077241 TI - Blebs and apoptotic bodies are B cell autoantigens. AB - Mounting evidence suggests that systemic lupus erythematosus autoantigens are derived from apoptotic cells. To characterize the potential interactions between apoptotic cells and B cells, the D56R/S76R variant of 3H9, a murine autoantibody that binds to DNA, chromatin, and anionic phospholipids, was compared with DNA4/1, a human anti-DNA autoantibody. Flow cytometry revealed that only D56R/S76R bound to Jurkat cells treated with either of three distinct proapoptotic stimuli, Ab binding was dependent on caspase activity, and immunoreactivity developed subsequent to annexin V binding. Confocal microscopy established a structural basis for the distinct kinetics of binding. D56R/S76R preferentially bound to membrane blebs of apoptotic cells, whereas annexin V binding did not require blebs. Inhibition of ROCK I kinase, an enzyme that stimulates nuclear fragmentation and fragment distribution into blebs, significantly reduced Ab binding. Because members of the collectin and pentraxin families of serum proteins bind to blebs on apoptotic cells and assist in the clearance of cellular remains, our results suggest that Abs to blebs could affect the recognition of apoptotic cells by cells of the innate immune system and thus modify tolerance to nuclear Ags. PMID- 12077242 TI - Human cytomegalovirus disrupts constitutive MHC class II expression. AB - CD8(+) and CD4(+) T lymphocytes are important in controlling human CMV (HCMV) infection, but the virus has evolved protean mechanisms to inhibit MHC-based Ag presentation and escape T lymphocyte immunosurveillance. Herein, the interaction of HCMV with the MHC class II Ag presentation pathway was investigated in cells stably transfected with class II transactivator. Flow cytometry experiments demonstrate that HCMV infection decreases cell-surface MHC class II expression. HCMV down-regulates MHC class II surface expression without a significant effect on class II RNA or steady-state protein levels. SDS-stability and confocal microscopy experiments demonstrate normal levels of steady-state peptide-loaded class II molecules in infected cells and that class II molecules reach late endosomal and HLA-DM positive peptide-loading compartments. However, MHC class II positive vesicles are retained in an abnormal perinuclear distribution. Finally, experiments with a mutant HCMV strain demonstrate that this novel mechanism of decreased MHC class II expression is not mediated by one of the known HCMV immunomodulatory genes. These defects in MHC class II expression combined with previously identified CMV strategies for decreasing MHC class I expression enables infected cells to evade T lymphocyte immunosurveillance. PMID- 12077243 TI - Expression and function of the Eph A receptors and their ligands ephrins A in the rat thymus. AB - Thymus development and function are dependent on the definition of different and graded microenvironments that provide the maturing T cell with the different signals that drive its maturation to a functional T lymphocyte. In these processes, cell-cell interactions, cell migration, and positioning are clues for the correct functioning of the organ. The Eph family of receptor tyrosine kinases and their ligands, the ephrins, has been implicated in all these processes by regulating cytoskeleton and adhesion functioning, but a systemic analysis of their presence and possible functional role in thymus has not yet been conducted. In this regard, the current study combines different experimental approaches for analyzing the expression of four members of the Eph A family and their ligands, ephrins A, in the embryonic and adult rat thymus. The patterns of Eph and ephrin expression in the distinct thymic regions were different but overlapping. In general, the studied Eph A were expressed on thymic epithelial cells, whereas ephrins A seem to be more restricted to thymocytes, although Eph A1 and ephrin A1 are expressed on both cell types. Furthermore, the supply of either Eph A-Fc or ephrin A-Fc fusion proteins to fetal thymus organ cultures interferes with T cell development, suggesting an important role for this family of proteins in the cell mechanisms that drive intrathymic T cell development. PMID- 12077244 TI - A defective NF-kappa B/RelB pathway in autoimmune-prone New Zealand black mice is associated with inefficient expansion of thymocyte and dendritic cells. AB - New Zeland Black (NZB) mice develop an autoimmune disease involving an abnormal B cell response to peripheral self Ags. This disease is associated with defects in other cell types and thymic stromal organization. We present evidence that NZB cells of various lineages, including thymocytes, fibroblasts, and dendritic precursor cells, show impaired proliferation and enhanced cell death in culture upon stimulation compared with non-autoimmune-prone mice such as C57BL/6. This phenotype explains the reduced efficiency of maturation of bone marrow-derived dendritic cells and the loss of TNF- or IL-1-dependent thymocyte costimulation. Upon TNF-induced activation of NZB thymocytes, nuclear translocation and DNA binding of RelA- and RelB-dependent NF-kappaB heterodimers are significantly reduced. This phenotype has a transcriptional signature, since the NZB, but not the nonobese diabetic, thymic transcriptome shows striking similarities with that of RelB-deficient thymuses. This partial NF-kappaB deficiency detected upon activation by proinflammatory cytokines could explain the disorganization of thymic microenvironments in NZB mice. These combined effects might reduce the efficiency of central tolerance and expose apoptotic debris generated during inflammatory processes to self recognition. PMID- 12077245 TI - Spatial raft coalescence represents an initial step in Fc gamma R signaling. AB - Characterization of lipid rafts as separated membrane microdomains consist of heterogeneous proteins suggesting that lateral assembly of rafts after Ag receptor cross-linking represents the earliest signal generating process. In line with the concept, cross-linked Ag receptors have been shown to associate with detergent-insoluble raft fraction without the aid of Src family kinases. However, it has not been established whether spatial raft coalescence could also precede Src family kinase activation. In this study, we showed that spatial raft coalescence after low-affinity FcgammaR cross-linking in RAW264.7 macrophages is independent of Src family kinase activity. The lateral raft assembly was found to be ascribed to the action of ligand-binding subunits, rather than to immunoreceptor tyrosine-based activation motif-bearing signal subunits, because monomeric murine FcgammaRIIb expressed in rat basophilic leukemia cells successfully induced spatial raft reorganization after cross-linking. We also showed that extracellular and transmembrane region of FcgammaRIIb is sufficient for raft stabilization. Moreover, this receptor fragment triggers rapid calcium mobilization and linker for activation of T cells phosphorylation, in a manner sensitive to Src family kinase inhibition and to cholesterol depletion. Presence of immunoreceptor tyrosine-based inhibitory motif and addition of immunoreceptor tyrosine-based activation motif to the receptor fragment abolished and enhanced the responses, respectively, but did not affect raft stabilization. These findings support the concept that ligand-binding subunit is responsible for raft coalescence, and that this event triggers initial biochemical signaling. PMID- 12077246 TI - Functional human T lymphocyte development from cord blood CD34+ cells in nonobese diabetic/Shi-scid, IL-2 receptor gamma null mice. AB - An experimental model for human T lymphocyte development from hemopoietic stem cells is necessary to study the complex processes of T cell differentiation in vivo. In this study, we report a newly developed nonobese diabetic (NOD)/Shi scid, IL-2Rgamma null (NOD/SCID/gamma(c)(null)) mouse model for human T lymphopoiesis. When these mice were transplanted with human cord blood CD34(+) cells, the mice reproductively developed human T cells in their thymus and migrated into peripheral lymphoid organs. Furthermore, these T cells bear polyclonal TCR-alphabeta, and respond not only to mitogenic stimuli, such as PHA and IL-2, but to allogenic human cells. These results indicate that functional human T lymphocytes can be reconstituted from CD34(+) cells in NOD/SCID/gamma(c)(null) mice. This newly developed mouse model is expected to become a useful tool for the analysis of human T lymphopoiesis and immune response, and an animal model for studying T lymphotropic viral infections, such as HIV. PMID- 12077247 TI - Characterization of the peptide-binding specificity of Mamu-B*17 and identification of Mamu-B*17-restricted epitopes derived from simian immunodeficiency virus proteins. AB - The SIV-infected rhesus macaque is an excellent model to examine candidate AIDS virus vaccines. These vaccines should elicit strong CD8(+) responses. Previous definition of the peptide-binding motif and optimal peptides for Mamu-A*01 has created a demand for Mamu-A*01-positive animals. We have now studied a second MHC class I molecule, Mamu-B*17, that is present in 12% of captive-bred Indian rhesus macaques. The peptide-binding specificity of the Mamu-B*17 molecule was characterized using single substitution analogs of two Mamu-B*17-binding peptides and libraries of naturally occurring sequences of viral or bacterial origin. Mamu B*17 uses position 2 and the C terminus of its peptide ligands as dominant anchor residues. The C terminus was found to have a very narrow specificity for the bulky aromatic residue W, with other aromatic residues (F and Y) being only occasionally tolerated. Position 2 is associated with a broad chemical specificity, readily accommodating basic (H and R), bulky hydrophobic (F and M), and small aliphatic (A) residues. Using this motif, we identified 50 peptides derived from SIV(mac)239 that bound Mamu-B*17 with an affinity of 500 nM or better. ELISPOT and intracellular cytokine-staining assays showed that 16 of these peptides were antigenic. We have, therefore, doubled the number of MHC class I molecules for which SIV-derived binding peptides have been characterized. This allows for the quantitation of immune responses through tetramers and analysis of CD8(+) function by intracellular cytokine-staining assays and ELISPOT. Furthermore, it is an important step toward the design of a multiepitope vaccine for SIV and HIV. PMID- 12077248 TI - NK cell receptors of the orangutan (Pongo pygmaeus): a pivotal species for tracking the coevolution of killer cell Ig-like receptors with MHC-C. AB - CD94, NKG2, Ly49, and killer cell Ig-like receptor (KIR) expressed by orangutan peripheral blood cells were examined by cloning and sequencing cDNA from a panel of individuals. Orthologs of human CD94, NKG2A, D, and F were defined. NKG2C and E are represented by one gene, Popy-NKG2CE, that is equidistant from the two human genes. Several Popy-CD94, NKG2A, and NKG2CE alleles were defined. Popy Ly49L is expressed in cultured NK cells and has a sequence consistent with it encoding a functional receptor. Orangutan KIR corresponding to the three KIR lineages expressed in humans and chimpanzees were defined. Popy-KIR2DL4 of lineage I is the only ortholog of a human or chimpanzee KIR, but in all individuals examined, the transcripts of this gene produced premature termination, either in the D2 domain or at the beginning of the cytoplasmic domain. Ten Popy-KIR3DL and one Popy-KIR3DS of lineage II are all closely related, but represent the products of at least two genes. The two Popy-KIR2DL and four Popy-KIR2DS of lineage III also represent two genes, both being more related to KIR2DS4 than to other human and chimpanzee KIR of lineage III. The Popy-KIR2D include ones predicted to be specific for the C1 epitope of MHC-C, but none specific for C2. This correlates with the observation that all orangutan MHC C allotypes examined have the C1 motif. PMID- 12077249 TI - A therapeutic CD4 monoclonal antibody inhibits TCR-zeta chain phosphorylation, zeta-associated protein of 70-kDa Tyr319 phosphorylation, and TCR internalization in primary human T cells. AB - The molecular mechanisms mediating the inhibitory effects of a humanized CD4 mAb YHB.46 on primary human CD4(+) T cells were investigated. Preincubation of T cells with soluble YHB.46 caused a general inhibition of TCR-stimulated protein tyrosine phosphorylation events, including a reduction in phosphorylation of p95(vav), linker for activation of T cells, and Src homology 2 domain-containing leukocyte protein of 76-kDa signaling molecules. A marked reduction in activation of the Ras/mitogen-activated protein kinase pathway was also observed. Examination of the earliest initiation events of TCR signal transduction showed that YHB.46 inhibited TCR-zeta chain phosphorylation together with recruitment and tyrosine phosphorylation of the zeta-associated protein of 70-kDa tyrosine kinase, particularly at Tyr(319), as well as reduced recruitment of p56(lck) to the TCR-zeta and zeta-associated protein of 70-kDa complex. These inhibitory events were associated with inhibition of TCR endocytosis. Our results show that the YHB.46 mAb is a powerful inhibitor of the early initiating events of TCR signal transduction. PMID- 12077250 TI - Genetic control of human NK cell repertoire. AB - Through differential killer cell Ig-like receptor (KIR) and CD94:NKG2 gene expression, human NK cells generate diverse repertoires, each cell having an inhibitory receptor for autologous HLA class I. Using a new method for measuring repertoire difference that integrates multiple flow cytometry parameters, we found individual repertoire stability, but population variability. Correlating repertoire differences with KIR and HLA genotype for 85 sibling pairs reveals the dominant influence of KIR genotype; HLA genotype having a subtle, modulating effect on relative KIR expression frequencies. HLA and/or KIR genotype also influences CD94:NKG2A expression. After HLA-matched stem cell transplantation, KIR repertoires either recapitulated that of the donor or were generally depressed for KIR expression. Human NK cell repertoires are defined by combinations of variable KIR and HLA class I genes and conserved CD94:NKG2 genes. PMID- 12077251 TI - Transcriptional regulation of GATA-3 by an intronic regulatory region and fetal liver zinc finger protein 1. AB - GATA-3 is a T cell-specific transcription factor and is essential for the development of the T cell lineage. The transcriptional regulation of GATA-3, however, remains elusive. In this study, we report the identification of a regulatory region located within the first intron of the murine GATA-3 gene. The intronic regulatory region contains both a positive and a negative cis-acting element but, as a whole, serves as a potent T cell-specific enhancer and is essential for the promoter activity in vitro. By using yeast one-hybrid screening, we discovered that fetal liver zinc finger protein 1 (Fliz1) could bind specifically to the negative cis-acting element, the sequence of which is conserved between the mouse and human GATA-3 genes. More importantly, overexpression of Fliz1 repressed the expression of GATA-3 in vivo and in vitro. Our data suggest that the expression of GATA-3 might be partly regulated by the intronic regulatory region and Fliz1 in a developmental stage-specific fashion. PMID- 12077252 TI - Recruitment of phosphatidylinositol 3-kinase to CD28 inhibits HIV transcription by a Tat-dependent mechanism. AB - Activation through the TCR and the costimulatory molecule CD28 influences the susceptibility of T cells to HIV-1 infection and regulates proviral gene expression. Signaling events initiated by CD28 that directly impact HIV-1 transcription have not been fully characterized. T cell lines expressing CD8alpha/28 chimeric receptors containing a mutation in tyrosine 173 to phenylalanine, which inhibits the recruitment of phosphatidylinositol 3-kinase (PI3K) to CD28, expressed higher levels of HIV-1 following T cell activation. Whereas constitutively active PI3K decreased provirus transcription, inhibiting endogenous PI3K with specific inhibitors or by overexpressing PTEN phosphatase enhanced HIV-1 expression. PI3K-dependent inhibition required the viral Tat protein and a trans activation response region element. Tat pull-down and coimmunoprecipitation experiments indicate that PI3K affects the formation of the Tat-associated kinase trans-activating complex. These studies demonstrate that PI3K negatively impacts HIV-1 transcription and that Tat activity is sensitive to T cell signaling events. PMID- 12077253 TI - Cloning and characterization of a promoter flanking the early B cell factor (EBF) gene indicates roles for E-proteins and autoregulation in the control of EBF expression. AB - The early B cell factor (EBF) is a transcription factor shown crucial for the development of B lymphocytes. The protein is expressed from the earliest stages of B cell development until the mature B cell stage, but the control elements responsible for the regulation of the gene are unknown. In this study, we report of the identification of a promoter region flanking the EBF gene. Several transcription start sites were identified by primer extension analysis in a region approximately 3.1 kb from the predicted ATG. Transient transfections revealed that this region was able to stimulate transcription of a reporter gene in B lymphoid and to a lesser extent, myeloid cells, but not in a pre-T cell line. The promoter was also able to functionally interact with E47, suggesting that the EBF gene may be a direct target for activation by E-proteins. In addition, functional binding of EBF to its own promoter was confirmed by EMSA and transfection assays indicating that the EBF protein may be involved in an autoregulatory loop. Finally, a tissue-restricted factor was able to bind an upstream regulatory region in B-lineage cells, further supporting the idea that the cloned promoter participates in the regulation of stage and lineage specific expression of the EBF gene. PMID- 12077254 TI - Unraveling of the polymorphic C lambda 2-C lambda 3 amplification and the Ke+Oz- polymorphism in the human Ig lambda locus. AB - Two polymorphisms of the human Ig(lambda) (IGL) locus have been described. The first polymorphism concerns a single, 2- or 3-fold amplification of 5.4 kb of DNA in the C(lambda)2-C(lambda)3 region. The second polymorphism is the Mcg( )Ke(+)Oz(-) isotype, which has only been defined via serological analyses in Bence-Jones proteins of multiple myeloma patients and was assumed to be encoded by a polymorphic C(lambda)2 segment because of its high homology with the Mcg( )Ke(-)Oz(-) C(lambda)2 isotype. It has been speculated that the Mcg(-)Ke(+)Oz(-) isotype might be encoded by a C(lambda) gene segment of the amplified C(lambda)2 C(lambda)3 region. We now unraveled both IGL gene polymorphisms. The amplification polymorphism appeared to result from a duplication, triplication, or quadruplication of a functional J-C(lambda)2 region and is likely to have originated from unequal crossing over of the J-C(lambda)2 and J-C(lambda)3 region via a 2.2-kb homologous repeat. The amplification polymorphism was found to result in the presence of one to five extra functional J-C(lambda)2 per genome regions, leading to decreased Ig(kappa):Ig(lambda) ratios on normal peripheral blood B cells. Via sequence analysis, we demonstrated that the Mcg(-)Ke(+)Oz(-) isotype is encoded by a polymorphic C(lambda)2 segment that differs from the normal C(lambda)2 gene segment at a single nucleotide position. This polymorphism was identified in only 1.5% (2 of 134) of individuals without J-C(lambda)2 amplification polymorphism and was not found in the J-C(lambda)2 amplification polymorphism of 44 individuals, indicating that the two IGL gene polymorphisms are not linked. PMID- 12077255 TI - Key residues of a major cytochrome P4502D6 epitope are located on the surface of the molecule. AB - Eukaryotically expressed CYP2D6 is the universal target of liver kidney microsomal Ab type 1 (LKM1) in both type 2 autoimmune hepatitis (AIH) and chronic hepatitis C virus (HCV) infection. In contrast, reactivity to prokaryotically expressed CYP2D6 protein and synthetic peptides is significantly lower in HCV infection than in AIH. The aim of the present study was to characterize LKM1 reactivity against a panel of eukaryotically expressed CYP2D6 constructs in the two conditions. LKM1-positive sera obtained from 16 patients with AIH and 16 with HCV infection were used as probes to perform a complete epitope mapping of CYP2D6. Reactivity to the full-length protein and 16 constructs thereof was determined by radioligand assay. We found that antigenicity is confined to the portion of the molecule C-terminal of aa 193, no reactivity being detectable against the aa sequence 1-193. Reactivity increases stepwise toward the C terminal in both AIH and HCV, but the frequency of reactivity in the two conditions differs significantly between aa 267-337. To further characterize this region, we introduced a five and a three amino acid swap mutation selected from the homologous regions of CYP2C9 and HCV. This maneuver resulted in a substantial loss of LKM1 binding in both conditions, suggesting that this region contains a major epitope. Molecular modeling revealed that CYP2D6(316-327) is exposed on the surface of the protein, and may represent a key target for the autoantibody. These findings provide an initial characterization of the antigenic constitution of the target of LKM1 in AIH and HCV infection. PMID- 12077256 TI - Negative feedback regulation of the tumor suppressor PTEN by phosphoinositide induced serine phosphorylation. AB - The PTEN tumor suppressor phosphatase directly counteracts the multiple functions of phosphatidylinositol 3-kinase by removing phosphate from the D3 position of inositol phospholipids. Like many lymphomas and leukemias, the Jurkat T cell line lacks PTEN protein due to frame-shift mutations in both PTEN alleles and therefore survives in long-term cell culture. We report that PTEN reintroduced into Jurkat was highly phosphorylated on serines 380 and 385 in its C terminus, particularly the former site. Phosphate was also detected at Ser(380) in PTEN in untransformed human T cells. Treatments that reduced the levels of D3 phospholipids in the cells resulted in reduced phosphorylation and accelerated degradation of PTEN. In contrast, expression of inactive PTEN-C124G or coexpression of a constitutively active protein kinase B led to increased phosphorylation and slower degradation of PTEN. These results suggest that PTEN normally is subjected to a feedback mechanism of regulation aimed at maintaining homeostatic levels of D3-phosphoinositides, which are crucial for T cell survival and activation. PMID- 12077257 TI - TCR engagement induces proline-rich tyrosine kinase-2 (Pyk2) translocation to the T cell-APC interface independently of Pyk2 activity and in an immunoreceptor tyrosine-based activation motif-mediated fashion. AB - The relocation of kinases in T lymphocytes during their cognate interaction with APCs is essential for lymphocyte activation. We found that the proline-rich tyrosine kinase-2 (Pyk2) is rapidly translocated to the T cell-APC contact area upon T cell-specific recognition of superantigen-pulsed APCs. Stimulation with anti-CD3-coated latex microspheres was sufficient for Pyk2 reorientation, and the coengagement of CD28 boosted Pyk2 redistribution. Nevertheless, Pyk2 translocation did not result in its recruitment to lipid rafts. Two results support that Pyk2 translocation was independent of its kinase activity. First, Lck activity was required for TCR-induced Pyk2 translocation, but not for TCR induced Pyk2 activation. Second, a kinase-dead Pyk2 mutant was equally translocated upon TCR triggering. In addition, Lck activity alone was insufficient to induce Pyk2 reorientation and activation, requiring the presence of at least one intact immunoreceptor tyrosine-based activation motif (ITAM). Despite the dependence on functional Lck and on phosphorylated ITAM for Pyk2 translocation, the ITAM-binding tyrosine kinase zeta-associated protein 70 (ZAP 70) was not essential. All these data suggest that, by translocating to the vicinity of the immune synapse, Pyk2 could play an essential role in T cell activation and polarized secretion of cytokines. PMID- 12077258 TI - Essential role of extrathymic T cells in protection against malaria. AB - Athymic nude mice carry neither conventional T cells nor NKT cells of thymic origin. However, NK1.1(-)TCR(int) cells are present in the liver and other immune organs of athymic mice, because these lymphocyte subsets are truly of extrathymic origin. In this study, we examined whether extrathymic T cells had the capability to protect mice from malarial infection. Although B6-nu/nu mice were more sensitive to malaria than control B6 mice, these athymic mice were able to survive malaria when a reduced number of parasitized erythrocytes (5 x 10(3) per mouse) were injected. At the fulminant stage, lymphocytosis occurred in the liver and the major expanding lymphocytes were NK1.1(-)TCR(int) cells (IL 2Rbeta(+)TCRalphabeta(+)). Unconventional CD8(+) NKT cells (V(alpha)14(-)) also appeared. Similar to the case of B6 mice, autoantibodies (IgM type) against denatured DNA appeared during malarial infection. Immune lymphocytes isolated from the liver of athymic mice which had recovered from malaria were capable of protecting irradiated euthymic and athymic mice from malaria when cell transfer experiments were conducted. In conjunction with the previous results in euthymic mice, the present results in athymic mice suggest that the major lymphocyte subsets associated with protection against malaria might be extrathymic T cells. PMID- 12077259 TI - Altered neutrophil trafficking during sepsis. AB - In sepsis, dysregulation of the inflammatory system is well known, as reflected in excessive inflammatory mediator production, complement activation, and appearance of defects in phagocytic cells. In the current study sepsis was induced in rats by cecal ligation/puncture. Early in sepsis the beta(1) and beta(2) integrin content on blood neutrophils increased in a nontranscriptional manner, and the increase in beta(2), but not beta(1), integrin content was C5a dependent. Similar changes could be induced in vitro on blood neutrophils following contact with phorbol ester or C5a. Direct injury of lungs of normal rats induced by deposition of IgG immune complexes (IgG-IC) caused 5-fold increases in the myeloperoxidase content that was beta(2), but not beta(1), dependent. In contrast, in cecal ligation/puncture lungs myeloperoxidase increased 10-fold after IgG immune complex deposition and was both beta(1) and beta(2) integrin dependent. These data suggest that sepsis causes enhanced neutrophil trafficking into the lung via mechanisms that are not engaged in the nonseptic state. PMID- 12077260 TI - Phenotype and regulation of persistent intracerebral T cells in murine Toxoplasma encephalitis. AB - Toxoplasma gondii is a parasite causing asymptomatic, persistent encephalitis. Protective CD4 and CD8 T cells are recruited to and accumulate in the brain in acute Toxoplasma encephalitis (TE), with slowly decreasing numbers in chronic TE. It is unclear how the size of the intracerebral T cell pool is regulated. Conceivably, permanent recruitment, proliferation, and apoptosis may be involved. We observed that in murine TE recruitment of T cells to the brain was terminated in chronic TE. In vivo 5-bromo-2'-deoxyuridine incorporation and in vitro T cell proliferation experiments revealed that intracerebral T cells did not proliferate, which was explained by the expression of the cell cycle inhibitors p21(Waf/cip1) and p27(Kip1) and the inhibitory activity of intracerebral F4/80(+) cells. TUNEL staining detected apoptotic T cells at low frequency corresponding to an increased expression of the anti-apoptotic molecules Bcl-2 and Bcl-x(L) and a reduced expression of the pro-apoptotic molecules Bad, Bax, and Fas ligand in CD4 and CD8 T cells. During progression from acute to chronic TE, both CD4 and CD8 T cells down-regulated CD45RB expression and expressed a differential pattern of cytokines. From these experiments it is concluded that the number of intracerebral T cells increases by recruitment of T cells during acute infection, whereas proliferation of intracerebral T cells does not play a role. In chronic TE, T cell recruitment is terminated, the phenotype of intracerebral T cells changes, and their number is gradually downsized by low level apoptosis, which, however, does not completely resolve the T cell infiltrates. PMID- 12077261 TI - Contribution of IL-18 to Th1 response and host defense against infection by Mycobacterium tuberculosis: a comparative study with IL-12p40. AB - The present study was conducted to critically determine the protective role of IL 18 in host response to Mycobacterium tuberculosis infection. IL-18-deficient (knockout (KO)) mice were slightly more prone to this infection than wild-type (WT) mice. Sensitivity of IL-12p40KO mice was lower than that of IL-12p40/IL-18 double KO mice. IFN-gamma production caused by the infection was significantly attenuated in IL-18KO mice compared with WT mice, as indicated by reduction in the levels of this cytokine in sera, spleen, lung, and liver, and its synthesis by spleen cells restimulated with purified protein derivatives. Serum IL-12p40 level postinfection and its production by peritoneal exudate cells stimulated with live bacilli were also significantly lower in IL-18KO mice than WT mice, suggesting that attenuated production of IFN-gamma was secondary to reduction of IL-12 synthesis. However, this was not likely the case, because administration of excess IL-12 did not restore the reduced IFN-gamma production in IL-18KO mice. In further studies, IL-18 transgenic mice were more resistant to the infection than control littermate mice, and serum IFN-gamma level and its production by restimulated spleen cells were increased in the former mice. Taken together, our results indicate that IL-18 plays an important role in Th1 response and host defense against M. tuberculosis infection although the contribution was not as profound as that of IL-12p40. PMID- 12077262 TI - Induction of CD1-restricted immune responses in guinea pigs by immunization with mycobacterial lipid antigens. AB - Group 1 CD1 molecules have been shown to present lipid and glycolipid Ags of mycobacteria to human T cells. However, a suitable animal model for the investigation of this component of antimycobacterial immunity has not yet been established. Previously, we found that guinea pigs express multiple isoforms of group 1 CD1 proteins that are homologous to human CD1b and CD1c. In this study, we show that CD1-restricted T cell responses can be generated in guinea pigs following immunization with lipid Ags from Mycobacterium tuberculosis. Splenic T cells from lipid Ag-immunized guinea pigs showed strong proliferative responses to total lipid Ags and partially purified glycolipid fractions from M. tuberculosis. These lipid Ag-reactive T cells were enriched in CD4-negative T cell fractions and showed cytotoxic activity against CD1-expressing guinea pig bone marrow-derived dendritic cells pulsed with M. tuberculosis lipid Ags. Using guinea pig cell lines transfected with individual CD1 isoforms as target cells in cytotoxic T cell assays, we found that guinea pig CD1b and CD1c molecules presented M. tuberculosis glycolipid Ags to T cells raised by mycobacterial lipid immunization. These results were confirmed using a T cell line derived from M. tuberculosis lipid Ag-immunized guinea pigs, which also showed CD1-restricted responses and cytolytic activity. Our results demonstrate that CD1-restricted responses against microbial glycolipid Ags can be generated in vivo by specific immunization and provide support for the use of the guinea pig as a relevant small animal model for the study of CD1-restricted immune responses to mycobacterial pathogens. PMID- 12077264 TI - Established human papillomavirus type 16-expressing tumors are effectively eradicated following vaccination with long peptides. AB - Peptide-based vaccines aimed at the induction of effective T cell responses against established cancers have so far only met with limited clinical success and clearly need to be improved. In a preclinical model of human papillomavirus (HPV)16-induced cervical cancer we show that prime-boost vaccinations with the HPV16-derived 35 amino-acid long peptide E7(43-77), containing both a CTL epitope and a Th epitope, resulted in the induction of far more robust E7-specific CD8(+) T cell responses than vaccinations with the minimal CTL epitope only. We demonstrate that two distinct mechanisms are responsible for this effect. First, vaccinations with the long peptide lead to the generation of E7-specific CD4(+) Th cells. The level of the induced E7-specific CD8(+) T cell response proved to be dependent on the interactions of these Th cells with professional APC. Second, we demonstrate that vaccination with the long peptide and dendritic cell activating agents resulted in a superior induction of E7-specific CD8(+) T cells, even when T cell help was excluded. This suggests that, due to its size, the long peptide was preferably endocytosed, processed, and presented by professional APCs. Moreover, the efficacy of this superior HPV-specific T cell induction was demonstrated in therapeutic prime-boost vaccinations in which the long peptide admixed with the dendritic cell-activating adjuvant oligodeoxynucleotide-CpG resulted in the eradication of large, established HPV16-expressing tumors. Because the vaccine types used in this study are easy to prepare under good manufacturing practice conditions and are safe to administer to humans, these data provide important information for future clinical trials. PMID- 12077263 TI - The role of proinflammatory cytokines in wasting disease during lymphocytic choriomeningitis virus infection. AB - Infection with pathogens often leads to loss of body weight, but the cause of weight loss during infection is poorly understood. We used the infection of mice with lymphocytic choriomeningitis virus (LCMV) as a model to study how pathogens induce weight loss. If LCMV is introduced into the CNS of CTL-deficient mice, the immune response against the virus leads to a severe weight loss called wasting disease. We planned to determine what components of this antiviral immune response mediate wasting disease. By adoptive transfer, we show that CD4 T cells activated by LCMV infection are sufficient to cause wasting disease. We examined the role of cytokines in LCMV-induced wasting disease using mice lacking specific cytokines or cytokine receptors. Results of adoptive transfer experiments suggest that TNF-alpha is not involved in LCMV-induced wasting disease and show that IFN gamma contributes to the disease. Consistent with a role for IFN-gamma in wasting, we find that IFN-gamma is necessary for LCMV-specific CD4 T cell responses in the CNS, most likely because it is required to induce MHC class II expression. Our data also indicate that IL-1 is required for LCMV-induced wasting and that IL-6 contributes to the wasting disease. Additionally, our results identify alpha-melanocyte-stimulating hormone as a potential mediator of the disease. Overall, this work defines the critical role of virus-primed CD4 T cells and of proinflammatory cytokines in the pathogenesis of wasting disease induced by LCMV infection. PMID- 12077265 TI - The murine cytomegalovirus immunomodulatory gene m152 prevents recognition of infected cells by M45-specific CTL but does not alter the immunodominance of the M45-specific CD8 T cell response in vivo. AB - Although in vitro studies have shown that herpesviruses, including murine CMV (MCMV), encode genes that interfere with the MHC class I pathway, their effects on the CTL response in vivo is unclear. We identified a D(b)-restricted CTL epitope from MCMV M45 by screening an MCMV genomic library using CTL clones isolated from mice infected with MCMV lacking m152. Because m152 severely inhibits CTL recognition of M45 in vitro, we questioned whether an M45-specific response would be generated in mice infected with wild-type MCMV expressing m152. Mice infected with wild-type MCMV or MCMVDelta(m)152 made similar responses to the M45 Ag. Moreover, we saw no skewing of the proportion of M45-specific CD8 T cells within the total MCMV-specific response after infection with MCMV with m152. Despite the profound effect m152 has on presentation of M45 in vitro, it does not affect the immunodominance of M45 in the CTL response in vivo. PMID- 12077266 TI - Selective expression of type I IFN genes in human dendritic cells infected with Mycobacterium tuberculosis. AB - Type I IFN regulates different aspects of the immune response, inducing a cell mediated immunity. We have recently shown that the infection of dendritic cells (DC) with Mycobacterium tuberculosis (Mtb) induces IFN-alpha. In this work we have monitored a rapid induction of IFN-beta followed by the delayed production of the IFN-alpha1 and/or -alpha13 subtypes. The Mtb infection rapidly activates the NF-kappaB complex and stimulates the phosphorylation of IFN regulatory factor (IRF)-3, events known to induce IFN-beta expression in viral infection. In turn, the autocrine production of IFN-beta induces the IFN-stimulated genes that contain binding sites for activated STATs in their promoters. Among the IFN stimulated genes induced in DC through STAT activation are IRF-1 and IRF-7. The expression of IRF-1 appears to be dependent on the sequential activation of NF kappaB and STAT-1. Once expressed, IRF-1 may further stimulate the transcription of IFN-beta. Induction of IRF-7 is also regulated at the transcriptional level through the binding of phosphorylated STAT-1 and STAT-2, forming the IFN stimulated gene factor-3 complex. In turn, the IRF-1 and IRF-7 expression appears to be required for the delayed induction of the IFN-alpha1/13 genes. Although correlative, our results strongly support the existence of a cascade of molecular events in Mtb-infected DC. Upon infection, constitutively expressed NF-kappaB and IRF-3 are activated and likely contribute to the rapid IFN-beta expression. In turn, IFN-beta-induced IRF-1 and IRF-7 may cooperate toward induction of IFN alpha1/13 if infection persists and these factors are activated. PMID- 12077268 TI - Anti-IL-10 therapeutic strategy using the immunomodulator AS101 in protecting mice from sepsis-induced death: dependence on timing of immunomodulating intervention. AB - The role of IL-10 in experimental sepsis is controversial. The nontoxic immunomodulator, ammonium trichloro(dioxoethylene-o,o')tellurate (AS101) has been previously shown to inhibit IL-10 expression at the transcriptional level. In this study, we show that in mice subjected to cecal ligation and puncture (CLP), treatment with AS101 12 h after, but not before, CLP significantly increased survival of septic mice. This was associated with a significant decrease in serum IL-10 and in IL-10 secretion by peritoneal macrophages 24-48 h after CLP. At that time, the ability of these cells to secrete TNF-alpha and IL-1beta was restored in AS101-treated mice. The increased survival of AS101-treated mice was due to the inhibition of IL-10, since cotreatment with murine rIL-10 abolished the protective activity of AS101. AS101 increased class II Ag expression on peritoneal macrophages, severely depressed in control mice, while it did not affect the expression of class I Ags. This was accompanied by a significant elevation in the level of IFN-gamma secreted by splenocytes. Moreover, AS101 ameliorated bacterial clearance in the peritoneum and blood and decreased severe multiple organ damage, as indicated by clinical chemistry. Furthermore, myeloperoxidase levels in the liver and lung of AS101-treated mice, an indirect means of determining the recruitment of neutrophils, were significantly decreased. We suggest that nontoxic agents such as AS101, with the capacity to inhibit IL-10 and stimulate macrophage functions, may have clinical potential in the treatment of sepsis, provided they are administered during the phase of sepsis characterized by immune suppression. PMID- 12077267 TI - Type I IFN as a natural adjuvant for a protective immune response: lessons from the influenza vaccine model. AB - The identification of natural adjuvants capable of selectively promoting an efficient immune response against infectious agents would represent an important advance in immunology, with direct implications for vaccine development, whose progress is generally hampered by the difficulties in defining powerful synthetic adjuvants suitable for clinical use. Here, we demonstrate that endogenous type I IFN is necessary for the Th1 type of immune response induced by typical adjuvants in mice and that IFN itself is an unexpectedly powerful adjuvant when administered with the human influenza vaccine, for inducing IgG2a and IgA production and conferring protection from virus challenge. The finding that these cytokines, currently used in patients, are necessary for full expression of adjuvant activity and are sufficient for the generation of a protective immune response opens new perspectives in understanding the basis of immunity and in vaccine development. PMID- 12077269 TI - The epistatic interrelationships of IL-1, IL-1 receptor antagonist, and the type I IL-1 receptor. AB - Mice lacking the gene for the IL-1R antagonist (IL-1ra) show abnormal development and homeostasis as well as altered responses to infectious and inflammatory stimuli. A reduction in the level of IL-1 signaling, either by deletion of the receptor or increased expression of IL-1ra, does not affect development or homeostasis, but does alter immune responses. In this study we use genetic epistasis to investigate the interdependence of selected genes in the IL-1 family in the regulation of these developmental and immunological processes. Deletion of the gene encoding the type I IL-1R (IL-1RI) is epistatic to deletion of the IL 1ra gene. Therefore, all functions of IL-1ra depend upon the presence of a functional receptor; there is no other target. Similarly, overexpression of the mRNA encoding the secreted form of IL-1ra is epistatic to deletion of the receptor antagonist, leaving the role of the intracellular splice variants of IL 1ra unknown. The abnormal development of IL-1ra-deficient mice is probably due to chronic overstimulation of the proinflammatory pathway via IL-1, but a clear single pathological defect is not apparent. These results support the model that the only essential function of IL-1ra in both health and disease is competitive inhibition of the IL-1RI. PMID- 12077270 TI - Differential glucocorticoid enhancement of the cytokine-driven transcriptional activation of the human acute phase serum amyloid A genes, SAA1 and SAA2. AB - The human acute phase serum amyloid A (A-SAA) genes, SAA1 and SAA2, have a high degree of sequence identity that extends approximately 450 bp upstream of their transcription start sites. Each promoter contains analogously positioned functional binding sites for the transcription factors NF-kappaB and NF-IL6. In human HepG2 hepatoma cells transfected with SAA promoter luciferase reporter constructs, administration of IL-1 and IL-6, singly or in combination, induced SAA1 and SAA2 transcriptional readouts that were qualitatively indistinguishable. However, under induced conditions, the SAA2 promoter had a significant quantitative transcriptional advantage over the SAA1 promoter. The application of the synthetic glucocorticoid dexamethasone in the context of cytokine stimulation enhanced the transcriptional activity of the SAA1, but not the SAA2, promoter such that readout from the former became equivalent to that from the latter. A putative glucocorticoid response element (GRE) is present (between residues -208 and -194) only in the SAA1 gene; a similar sequence in the corresponding region of the SAA2 gene is disrupted by a nine-residue insertion. The SAA1 GRE was shown to be functionally active and the SAA2 disrupted GRE was shown to be functionally inactive in experiments using reporter constructs carrying SAA1 and SAA2 promoters that had been modified by site-specific mutagenesis. Quantitative analysis of transcript-specific RT-PCR products, derived from SAA1 and SAA2 mRNAs after treatment of HepG2 cells with cytokines in the presence or absence of dexamethasone, confirmed that the endogenous SAA1 gene has a cytokine-driven transcriptional disadvantage that is superseded by a marginal transcriptional advantage when glucocorticoids are present. PMID- 12077271 TI - Cooperative effects of Th2 cytokines and allergen on normal and asthmatic bronchial epithelial cells. AB - In sensitized individuals, exposure to allergens such as Dermatophagoides pteronyssinus (Der p) causes Th2 polarization and release of cytokines, including IL-4 and IL-13. Because Der p extracts also have direct effects on epithelial cells, we hypothesized that allergen augments the effects of Th2 cytokines by promoting mediator release from the bronchial epithelium in allergic asthma. To test our hypothesis, primary bronchial epithelial cultures were grown from bronchial brushings of normal and atopic asthmatic subjects. RT-PCR showed that each culture expressed IL-4R(alpha), common gamma-chain, and IL-13R(alpha)(1), as well as IL-13R(alpha)(2), which negatively regulates IL-13 signaling; FACS analysis confirmed IL-13R(alpha)(2) protein expression. Exposure of epithelial cultures to either Der p extracts, TNF-alpha, IL-4, or IL-13 enhanced GM-CSF and IL-8 release, and this was partially suppressible by corticosteroids. Simultaneous exposure of the epithelial cultures to IL-4 or IL-13 together with Der p resulted in a further increase in cytokine release, which was at least additive. Release of TGF-alpha was also increased by TNF-alpha and combinations of IL-4, IL-13, and Der p; however, this stimulation was only significant in the asthma-derived cultures. These data suggest that, in an allergic environment, Th2 cytokines and allergen have the potential to sustain airway inflammation through a cooperative effect on cytokine release by the bronchial epithelium. Our novel finding that IL-4, IL-13, and allergen enhance release of TGF-alpha, a ligand for the epidermal growth factor receptor that stimulates fibroblast proliferation and goblet cell differentiation, provides a potential link between allergen exposure, Th2 cytokines, and airway remodelling in asthma. PMID- 12077272 TI - Modulation of Mac-1 (CD11b/CD18)-mediated adhesion by the leukocyte-specific protein 1 is key to its role in neutrophil polarization and chemotaxis. AB - Leukocyte-specific protein 1 (LSP1) is an intracellular filamentous-actin binding protein which modulates cell motility. The cellular process in which LSP1 functions to regulate motility is not yet identified. In this study, we show that LSP1 negatively regulates fMLP-induced polarization and chemotaxis of neutrophils through its function on adhesion via specific integrins. Using LSP1-deficient (Lsp1(-/-)) mice, we show increased neutrophil migration into mouse knee joints during zymosan-induced acute inflammation, an inflammatory model in which the number of resident synoviocytes are not affected by LSP1-deficiency. In vitro chemotaxis experiments performed by time-lapse videomicroscopy showed that purified Lsp1(-/-) bone-marrow neutrophils exhibit an increased migration rate toward a gradient of fMLP as compared with wild-type neutrophils. This difference was observed when cells migrated on fibrinogen, but not fibronectin, suggesting a role for LSP1 in modulating neutrophil adhesion by specific integrins. LSP1 is also a negative regulator of fMLP-induced adhesion to fibrinogen or ICAM-1, but not to ICAM-2, VCAM-1, or fibronectin. These results suggest that LSP1 regulates the function of Mac-1 (CD11b/CD18), which binds only to fibrinogen and ICAM-1 among the substrates we tested. fMLP-induced filamentous actin polarization is also increased in the absence of LSP1 when cells were layered on fibrinogen, but not on fibronectin. Our findings suggest that the increased neutrophil recruitment in Lsp1(-/-) mice during acute inflammation derives from the negative regulatory role of LSP1 on neutrophil adhesion, polarization, and migration via specific integrins, such as Mac-1, which mediate neutrophil responses to chemotactic stimuli. PMID- 12077273 TI - Differing activities of homeostatic chemokines CCL19, CCL21, and CXCL12 in lymphocyte and dendritic cell recruitment and lymphoid neogenesis. AB - Despite their widespread expression, the in vivo recruitment activities of CCL19 (EBV-induced molecule 1 ligand chemokine) and CXCL12 (stromal cell-derived factor 1) have not been established. Furthermore, although CXCL13 (B lymphocyte chemoattractant) has been shown to induce lymphoid neogenesis through induction of lymphotoxin (LT)alpha1beta2, it is unclear whether other homeostatic chemokines have this property. In this work we show that ectopic expression in pancreatic islets of CCL19 leads to small infiltrates composed of lymphocytes and dendritic cells and containing high endothelial venules and stromal cells. Ectopic CXCL12 induced small infiltrates containing few T cells but enriched in dendritic cells, B cells, and plasma cells. Comparison of CCL19 transgenic mice with mice expressing CCL21 (secondary lymphoid tissue chemokine) revealed that CCL21 induced larger and more organized infiltrates. A more significant role for CCL21 is also suggested in lymphoid tissues, as CCL21 protein was found to be present in lymph nodes and spleen at much higher concentrations than CCL19. CCL19 and CCL21 but not CXCL12 induced LTalpha1beta2 expression on naive CD4 T cells, and treatment of CCL21 transgenic mice with LTbetaR-Fc antagonized development of organized lymphoid structures. LTalpha1beta2 was also induced on naive T cells by the cytokines IL-4 and IL-7. These studies establish that CCL19 and CXCL12 are sufficient to mediate cell recruitment in vivo and they indicate that LTalpha1beta2 may function downstream of CCL21, CCL19, and IL-2 family cytokines in normal and pathological lymphoid tissue development. PMID- 12077274 TI - Impaired IFN-gamma-dependent inflammatory responses in human keratinocytes overexpressing the suppressor of cytokine signaling 1. AB - Keratinocytes contribute relevantly to the pathogenesis of inflammatory skin diseases by expressing a variety of proinflammatory molecules, with T cell derived IFN-gamma being the most potent keratinocyte activator. Suppressor of cytokine signaling (SOCS)1 and SOCS3 are negative regulators of IFN-gamma signaling and are induced in many cell types by IFN-gamma itself or by other cytokines. We show in this work that SOCS1, SOCS2, SOCS3, and cytokine-inducible SH2-containing protein mRNA were up-regulated by IFN-gamma in normal human keratinocytes, whereas only SOCS1 or SOCS1 and cytokine-inducible SH2-containing protein were induced by TNF-alpha or IL-4, respectively. SOCS1, SOCS2, and SOCS3 proteins were undetectable in healthy skin and highly expressed in the epidermis of psoriasis and allergic contact dermatitis, but were only weakly expressed in atopic dermatitis skin. In keratinocytes transiently transfected with SOCS1 or SOCS3 the IFN-gamma-induced transactivation of an IFN-gamma-responsive reporter gene was markedly inhibited. SOCS1 and SOCS3 overexpression in keratinocyte stable clones inhibited IFN-gamma-induced phosphorylation of IFN-gammaR(alpha) and activation of STAT1 and STAT3. Furthermore, SOCS1 and, to a lesser extent, SOCS3 reduced membrane expression of ICAM-1 and HLA-DR, and release of IFN-gamma inducible protein-10, monokine induced by IFN-gamma, and monocyte chemoattractant protein-1 by keratinocyte clones promoted by IFN-gamma. SOCS1-expressing keratinocytes showed constitutively higher, but not IFN-gamma-inducible, IL-8 levels compared with SOCS2 and SOCS3 clones, and were resistant to IFN-gamma mediated growth inhibition. Targeting keratinocyte SOCS1 may represent a novel therapeutic approach to IFN-gamma-dependent skin diseases. PMID- 12077276 TI - Complement deficiency ameliorates collagen-induced arthritis in mice. AB - Collagen-induced arthritis (CIA) is an experimental animal model of human rheumatoid arthritis being characterized by synovitis and progressive destruction of cartilage and bone. CIA is induced by injection of heterologous or homologous collagen type II in a susceptible murine strain. DBA/1J mice deficient of complement factors C3 (C3(-/-)) and factor B (FB(-/-)) were generated to elucidate the role of the complement system in CIA. When immunized with bovine collagen type II emulsified in CFA, control mice developed severe arthritis and high CII-specific IgG Ab titers. In contrast, the C3(-/-) and FB(-/-) were highly resistant to CIA and displayed decreased CII-specific IgG Ab response. A repeated bovine collagen type II exposure 3 wk after the initial immunization led to an increase in the Ab response in all mice and triggered arthritis also in the complement-deficient mice. Although the arthritic score of the C3(-/-) mice was low, the arthritis in FB(-/-) mice ranked intermediate with regard to C3(-/-) and control mice. We conclude that complement activation by both the classical and the alternative pathway plays a deleterious role in CIA. PMID- 12077275 TI - New IL-17 family members promote Th1 or Th2 responses in the lung: in vivo function of the novel cytokine IL-25. AB - We have biologically characterized two new members of the IL-17 cytokine family: IL-17F and IL-25. In contrast to conventional in vitro screening approaches, we have characterized the activity of these new molecules by direct in vivo analysis and have compared their function to that of other IL-17 family members. Intranasal administration of adenovirus expressing IL-17, IL-17C, or IL-17F resulted in bronchoalveolar lavage neutrophilia and inflammatory gene expression in the lung. In contrast, intranasal administration of IL-25-expressing adenovirus or IL-25 protein resulted in the production of IL-4, IL-5, IL-13, and eotaxin mRNA in the lung and marked eosinophilia in the bronchoalveolar lavage and lung tissue. Mice given intranasal IL-25 also developed epithelial cell hyperplasia, increased mucus secretion, and airway hyperreactivity. IL-25 gene expression was detected following Aspergillus and Nippostrongylus infection in the lung and gut, respectively. IL-25-induced eosinophilia required IL-5 and IL 13, but not IL-4 or T cells. Following IL-25 administration, the IL-5(+) staining cells were CD45R/B220(+), Thy-1(+/-), but were NK1.1-, Ly-6G(GR-1)-, CD4-, CD3-, and c-kit-negative. gamma-common knockout mice did not develop eosinophilia in response to IL-25, nor were IL-5(+) cells detected. These findings suggest the existence of a previously unrecognized cell population that may initiate Th2-like responses by responding to IL-25 in vivo. Further, these data demonstrate the heterogeneity of function within the IL-17 cytokine family and suggest that IL-25 may be an important mediator of allergic disease via production of IL-4, IL-5, IL 13, and eotaxin. PMID- 12077277 TI - IL-15-dependent activation-induced cell death-resistant Th1 type CD8 alpha beta+NK1.1+ T cells for the development of small intestinal inflammation. AB - To clarify the role of IL-15 at local sites, we engineered a transgenic (Tg) mouse (T3(b)-IL-15 Tg) to overexpress human IL-15 preferentially in intestinal epithelial cells by the use of T3(b)-promoter. Although IL-15 was expressed in the entire small intestine (SI) and large intestines of the Tg mice, localized inflammation developed in the proximal SI only. Histopathologic study revealed reduced villus length, marked infiltration of lymphocytes, and vacuolar degeneration of the villus epithelium, beginning at approximately 3-4 mo of age. The numbers of CD8(+) T cells, especially CD8alphabeta(+) T cells expressing NK1.1, were dramatically increased in the lamina propria of the involved SI. The severity of inflammation corresponded to increased numbers of CD8alphabeta(+)NK1.1(+) T cells and levels of production of the Th1-type cytokines IFN-gamma and TNF-alpha. Locally overexpressed IL-15 was accompanied by increased resistance of CD8alphabeta(+) NK1.1(+) T cells to activation-induced cell death. Our results suggest that chronic inflammation in the SI in this murine model is mediated by dysregulation of epithelial cell-derived IL-15. The model may contribute to understanding the role of CD8(+) T cells in human Crohn's disease involving the SI. PMID- 12077278 TI - Proinflammatory cytokine IL-1 beta promotes tumor growth of Lewis lung carcinoma by induction of angiogenic factors: in vivo analysis of tumor-stromal interaction. AB - Inflammatory conditions are associated with tumor development. IL-1beta is a multifunctional and proinflammatory cytokine that affects nearly all types of cells. To investigate the role of IL-1beta in tumor growth in vivo, we transduced the retroviral vector coding human IL-1beta gene into mouse Lewis lung carcinoma (LLC) cells and subsequently inoculated the transformant (LLC/IL-1beta) to syngeneic C57BL/6 mice. Tumors derived from LLC/IL-1beta grew faster (240%, day 18, vs null-vector control LLC/neo; p < 0.01) and showed more abundant vasculature (250%, vs LLC/neo; p < 0.05), whereas LLC/IL-1beta cells, LLC/neo cells, and wild-type LLC cells did not show any significant difference in the growth rate in vitro. As compared with LLC/neo cells, LLC/IL-1beta cells secreted 2-fold the amount of vascular endothelial growth factor and >10-fold the amount of macrophage-inflammatory protein-2 (CXCL2), one of whose main functions is angiogenesis. Although LLC/IL-1beta itself did not secrete hepatocyte growth factor (HGF), the tumor derived from LLC/IL-1beta cells also contained a >4-fold higher concentration of HGF, another angiogenic factor. In situ hybridization of HGF mRNA in LLC/IL-1beta tumor sections demonstrated that stromal fibroblasts and infiltrating cells overexpressed HGF mRNA. Moreover, when cultured in the presence of HGF in vitro, LLC/IL-1beta cells secreted even larger amounts of vascular endothelial growth factor and macrophage-inflammatory protein-2. The antiangiogenic agent TNP-470 and anti-CXCR2 Ab inhibited the tumor growth of LLC/IL-1beta cells in vivo. These results indicated that secreting IL-1beta into the tumor milieu induces several angiogenic factors from tumor and stromal cells and thus promotes tumor growth through hyperneovascularization. PMID- 12077279 TI - Neutrophil transepithelial migration: evidence for sequential, contact-dependent signaling events and enhanced paracellular permeability independent of transjunctional migration. AB - Active migration of polymorphonuclear leukocytes (PMN) through the intestinal crypt epithelium is a hallmark of inflammatory bowel disease and correlates with patient symptoms. Previous in vitro studies have shown that PMN transepithelial migration results in increased epithelial permeability. In this study, we modeled PMN transepithelial migration across T84 monolayers and demonstrated that enhanced paracellular permeability to small solutes occurred in the absence of transepithelial migration but required both PMN contact with the epithelial cell basolateral membrane and a transepithelial chemotactic gradient. Early events that occurred before PMN entering the paracellular space included increased permeability to small solutes (<500 Da), enhanced phosphorylation of regulatory myosin L chain, and other as yet undefined proteins at the level of the tight junction. No redistribution or loss of tight junction proteins was detected in these monolayers. Late events, occurring during actual PMN transepithelial migration, included redistribution of epithelial serine-phosphorylated proteins from the cytoplasm to the nucleus in cells adjacent to migrating PMN. Changes in phosphorylation of multiple proteins were observed in whole cell lysates prepared from PMN-stimulated epithelial cells. We propose that regulation of PMN transepithelial migration is mediated, in part, by sequential signaling events between migrating PMN and the epithelium. PMID- 12077280 TI - A role for oxidative stress in apoptosis: oxidation and externalization of phosphatidylserine is required for macrophage clearance of cells undergoing Fas mediated apoptosis. AB - Exposure of phosphatidylserine (PS) on the surface of apoptotic cells has been suggested to serve as an important recognition signal for macrophages. In this work we show that triggering of the death receptor Fas on Jurkat cells results in the generation of reactive oxygen species with oxidation and externalization of PS but not of the other major aminophospholipid, phosphatidylethanolamine. These cells were readily ingested by several classes of macrophages, whereas Raji cells, which are defective for Fas-induced PS exposure, remained unengulfed. However, when Raji cells were incubated with the thiol-reactive agent N ethylmaleimide to induce PS exposure in the absence of other features of apoptosis, these cells were also engulfed by macrophages. Phagocytosis of Fas triggered Jurkat cells was inhibited by superoxide dismutase and catalase, which prevent oxidation of PS while allowing PS to remain externalized on these cells. Moreover, liposomes containing oxidized PS (PS-OX) were more potent inhibitors of phagocytosis than those containing its nonoxidized counterpart. Finally, enrichment of the plasma membrane of Jurkat or Raji cells, or myeloid leukemic HL 60 cells, with exogenous PS resulted in phagocytic cell clearance, and this process was further enhanced when PS was substituted for by PS-OX. Taken together, our data suggest that the presence of PS-OX in conjunction with nonoxidized PS on the cell surface is an important signal for macrophage clearance of apoptotic cells. PMID- 12077281 TI - Skin graft rejection elicited by beta 2-microglobulin as a minor transplantation antigen involves multiple effector pathways: role of Fas-Fas ligand interactions and Th2-dependent graft eosinophil infiltrates. AB - Beta(2)-microglobulin (beta(2)m)-derived peptides are minor transplantation Ags in mice as beta(2)m-positive skin grafts (beta(2)m(+/+)) are rejected by genetically beta(2)m-deficient recipient mice (beta(2)m(-/-)). We studied the effector pathways responsible for the rejection induced by beta(2)-microglobulin derived minor transplantation Ags. The rejection of beta(2)m(+/+) skin grafts by naive beta(2)m(-/-) mice was dependent on both CD4 and CD8 T cells as shown by administration of depleting mAbs. Experiments performed with beta(2)m(-/-)CD8(-/ ) double knockout mice grafted with a beta(2)m(+/+) MHC class I-deficient skin showed that sensitized CD4 T cells directed at beta(2)m peptides-MHC class II complexes are sufficient to trigger rapid rejection. Rejection of beta(2)m(+/+) grafts was associated with the production of IL-5 in vitro, the expression of IL 4 and IL-5 mRNAs in the grafted tissue, and the presence within rejected grafts of a considerable eosinophil infiltrate. Blocking IL-4 and IL-5 in vivo and depleting eosinophils with an anti-CCR3 mAb prevented graft eosinophil infiltration and prolonged beta(2)m(+/+) skin graft survival. Lymphocytes from rejecting beta(2)m(-/-) mice also displayed an increased production of IFN-gamma after culture with beta(2)m(+/+) minor alloantigens. In vivo neutralization of IFN-gamma inhibited skin graft rejection. Finally, beta(2)m(+/+) skin grafts harvested from B6(lpr/lpr) donor mice, which lack a functional Fas molecule, survived longer than wild-type beta(2)m(+/+) skin grafts, showing that Fas-Fas ligand interactions are involved in the rejection process. We conclude that IL-4- and IL-5-dependent eosinophilic rejection, IFN-gamma-dependent mechanisms, and Fas-Fas ligand interactions are effector pathways in the acute rejection of minor transplantation Ags. PMID- 12077282 TI - Inhibition of endogenous TGF-beta during experimental osteoarthritis prevents osteophyte formation and impairs cartilage repair. AB - Osteoarthritis has as main characteristics the degradation of articular cartilage and the formation of new bone at the joint edges, so-called osteophytes. In this study enhanced expression of TGF-beta1 and -beta3 was detected in developing osteophytes and articular cartilage during murine experimental osteoarthritis. To determine the role of endogenous TGF-beta on osteophyte formation and articular cartilage, TGF-beta activity was blocked via a scavenging soluble TGF-beta-RII. Our results clearly show that inhibition of endogenous TGF-beta nearly completely prevented osteophyte formation. In contrast, treatment with recombinant soluble TGF-beta-RII markedly enhanced articular cartilage proteoglycan loss and reduced the thickness of articular cartilage. In conclusion, we show for the first time that endogenous TGF-beta is a crucial factor in the process of osteophyte formation and has an important function in protection against cartilage loss. PMID- 12077283 TI - An autoreactive gamma delta TCR derived from a polymyositis lesion. AB - To investigate the role of gammadelta T cells in human autoimmune disease we expressed and characterized a gammadelta TCR from an autoimmune tissue lesion. The TCR was first identified in a rare form of polymyositis characterized by a monoclonal infiltrate of gammadelta T cells which invaded and destroyed skeletal muscle fibers. The Vgamma1.3-Jgamma1-Cgamma1/Vdelta2-Jdelta3 TCR cDNA of the original muscle invasive gammadelta T cell clone was reconstructed from unrelated cDNA and transfected into the mouse hybridoma BW58alpha(-)beta(-). Appropriate anti-human gammadelta TCR Abs stimulated the TCR transfectants to produce IL-2, thus demonstrating that the human gammadelta TCR functionally interacted with murine signaling components. The transfected Vgamma1.3/Vdelta2 TCR recognized a cytosolic protein expressed in cultured human myoblasts and TE671 rhabdomyosarcoma cells. The Ag was recognized in the absence of presenting cells. Using a panel of control gammadelta TCR transfectants with defined exchanges in different positions of both TCR chains, we showed that the gammadelta TCR recognized its Ag in a TCR complementarity-determining region 3-dependent way. To our knowledge, this is the first example of a molecularly defined gammadelta TCR directly derived from an autoimmune tissue lesion. The strategy used in this study may be applicable to other autoimmune diseases. PMID- 12077284 TI - Analysis of the innate and adaptive phases of allograft rejection by cluster analysis of transcriptional profiles. AB - Both clinical and experimental observations suggest that allograft rejection is a complex process with multiple components that are, at least partially, functionally redundant. Studies using graft recipients deficient in various genes including chemokines, cytokines, and other immune-associated genes frequently produce a phenotype of delayed, but not indefinitely prevented, rejection. Only a small subset of genetic deletions (for example, TCR(alpha) or beta, MHC I and II, B7-1 and B7-2, and recombinase-activating gene) permit permanent graft acceptance suggesting that rejection is orchestrated by a complex network of interrelated inflammatory and immune responses. To investigate this complex process, we have used oligonucleotide microarrays to generate quantitative mRNA expression profiles following transplantation. Patterns of gene expression were confirmed with real-time PCR data. Hierarchical clustering algorithms clearly differentiated the early and late phases of rejection. Self-organizing maps identified clusters of coordinately regulated genes. Genes up-regulated during the early phase included genes with prior biological functions associated with ischemia, injury, and Ag-independent innate immunity, whereas genes up-regulated in the late phase were enriched for genes associated with adaptive immunity. PMID- 12077285 TI - Antimelanoma activity of CTL generated from peripheral blood mononuclear cells after stimulation with autologous dendritic cells pulsed with melanoma gp100 peptide G209-2M is correlated to TCR avidity. AB - Anchor residue-modified peptides derived from tumor-associated Ag have demonstrated success in engendering immune responses in clinical studies. However, tumor regression does not always correlate with immune responses. One hypothesis to explain this is that CTL resulting from such immunization approaches are variable in antitumor potency. In the present study, we evaluated this hypothesis by characterizing the activity of tumor-associated Ag-specific CTL. We chose an anchor residue-modified peptide from gp100, G209-2M, and used peptide-pulsed dendritic cells to generate CTL from PBMC of HLA-A2(+) normal donors. The specificities and avidities of the resulting CTL were evaluated. The results demonstrate that CTL generated by G209-2M can be classified into three categories: G209-2M-specific CTL which are cytotoxic only to G209-2M-pulsed targets; peptide-specific CTL which recognize both G209 and G209-2M peptides but not melanomas; and melanoma-reactive CTL which recognize peptide-pulsed targets as well as HLA-A2(+)gp100(+) melanomas. CTL that kill only peptide-pulsed targets require a higher peptide concentration to mediate target lysis, whereas CTL that lyse melanomas need a lower peptide concentration. Increasing peptide density on melanomas by loading exogenous G209 peptide enhances their sensitivity to peptide specific CTL. High avidity CTL clones also demonstrate potent antimelanoma activity in melanoma model in nude mice. Injection of G209 peptide around transplanted tumors significantly enhances the antitumor activity of low avidity CTL. These results suggest that peptide stimulation causes expansion of T cell populations with a range of avidities. Successful immunotherapy may require selective expansion of the higher-avidity CTL and intratumor injection of the peptide may enhance the effect of peptide vaccines. PMID- 12077286 TI - Identification of HLA-A*0201-restricted T cell epitopes derived from the novel overexpressed tumor antigen calcium-activated chloride channel 2. AB - Vaccination against tumor Ags may become a promising treatment modality especially in cancer types where other therapeutic approaches fail. However, diversity of tumors requires that a multitude of Ags become available. Differential expression in normal vs cancerous tissues, both at the mRNA and the protein level, may identify Ag candidates. We have previously compared transcripts from squamous cell lung cancer and normal lung tissue using differential display analysis, and found a transcript that was overexpressed in malignant cells and was identical with the calcium-activated chloride channel 2 (CLCA2) gene. We have now selected HLA-A2-restricted peptides from CLCA2, and have generated T cell lines against the CLCA2-derived KLLGNCLPTV, LLGNCLPTV, and SLQALKVTV peptides using in vitro priming. Specificity of T cells was ascertained in ELISPOT assays. The primed T cells also recognized allogeneic tumor cells in an Ag-specific and HLA-restricted fashion. Moreover, peptide LLGNCLPTV was also independently recognized by CD8(+) T cells expanded from pancreatic carcinoma/T cell cocultures. CLCA2-specific CD8(+) T cells were absent from the peripheral blood of healthy donors. These data indicate that an immune response can be induced against CLCA2, which thus may become an important Ag for anti-tumor vaccination approaches. PMID- 12077287 TI - High immunogenicity of intracellular myelin oligodendrocyte glycoprotein epitopes. AB - Multiple sclerosis (MS) is an inflammatory and demyelinating disease of the CNS with associated axonal loss. There is strong evidence for an autoimmune pathogenesis driven by myelin-specific T cells. Myelin oligodendrocyte glycoprotein (MOG) induces a type of experimental autoimmune encephalomyelitis in animals which is very MS-like since there are demyelinating CNS lesions and axonal loss. This underscores the potential role of MOG in MS pathogenesis. We performed a T cell reactivity pattern analysis of MS patients at the onset of relapse or progression of neurological deficits and controls that were stratified for the genetic risk factor HLA-DRB1*1501. For the first time, we show that there is an HLA-DR-restricted promiscuous dominant epitope for CD4(+) T cells within the transmembrane/intracellular part of MOG comprising aa 146-154 (FLCLQYRLR). Surprisingly, controls had broader T cell reactivity patterns toward MOG peptides compared with MS patients, and the transmembrane and intracellular parts of MOG were much more immunogenic compared with the extracellular part. Measurements of in vitro binding affinities revealed that HLA-DRB1*1501 molecules bound MOG 146 154 with intermediate and HLA-DRB1*0401 molecules with weak affinities. The binding of MOG 146-154 was comparable or better than myelin basic protein 85-99, which is the dominant myelin basic protein epitope in context with HLA-DRB1*1501 molecules in MS patients. This is the first study in which the data underscore the need to investigate the pathogenic or regulatory role of the transmembrane and intracellular part of MOG for MS in more detail. PMID- 12077288 TI - Identification of naturally processed CD4 T cell epitopes from the prostate specific antigen kallikrein 4 using peptide-based in vitro stimulation. AB - Kallikrein (KLK)4 is a recently described member of the tissue kallikrein gene family that is specifically expressed in normal and prostate tumor tissues. The tissue-specific expression profile of this molecule suggests that it might be useful as a vaccine candidate against prostate cancer. To examine the presence of CD4 T cells specific for KLK4 in PBMC of normal individuals, a peptide-based in vitro stimulation protocol was developed that uses overlapping KLK4-derived peptides spanning the majority of the KLK4 protein. Using this methodology, three naturally processed CD4 epitopes derived from the KLK4 sequence are identified. These epitopes are restricted by HLA-DRB1*0404, HLA-DRB1*0701, and HLA-DPB1*0401 class II alleles. CD4 T cell clones specific for these epitopes are shown to efficiently and specifically recognize both recombinant KLK4 protein and lysates from prostate tumor cell lines virally infected to express KLK4. CD4 T cells specific for these KLK4 epitopes are shown to exist in PBMC from multiple male donors that express the relevant class II alleles, indicating that a CD4 T cell repertoire specific for KLK4 is present and potentially expandable in prostate cancer patients. The demonstration that KLK4-specific CD4 T cells exist in the peripheral circulation of normal male donors and the identification of naturally processed KLK4-derived CD4 T cell epitopes support the use of KLK4 in whole gene , protein-, or peptide-based vaccine strategies against prostate cancer. Furthermore, the identification of naturally processed KLK4-derived epitopes provides valuable tools for monitoring preexisting and vaccine-induced responses to this molecule. PMID- 12077289 TI - Differential induction of mucosal and systemic antibody responses in women after nasal, rectal, or vaginal immunization: influence of the menstrual cycle. AB - A cholera vaccine containing killed vibrios and cholera toxin B subunit (CTB) was used to compare mucosal immunization routes for induction of systemic and mucosal Ab. Four groups of women were given three monthly immunizations by the rectal immunization (R(imm)) route, nasal immunization (N(imm)) route, or vaginal immunization route during either the follicular (V-FP(imm)) or luteal (V-LP(imm)) menstrual cycle phase. N(imm) was performed with 10-fold less vaccine to determine if administration of less Ag by this route can, as in rodents, produce mucosal Ab responses comparable to those induced by higher dose R(imm) or vaginal immunization. Concentrations of Ab induced in sera and secretions were measured by ELISA. None of these routes produced durable salivary Ab responses. N(imm) induced greatest levels of CTB-specific IgG in sera. R(imm) failed to generate CTB-specific IgA in genital tract secretions. N(imm), V-FP(imm), and V-LP(imm) all produced cervical CTB-specific IgA responses comparable in magnitude and frequency. However, only V-FP(imm) induced cervical IgA2-restricted Ab to the bacterial LPS vaccine component. V-FP(imm), but not V-LP(imm), also induced CTB specific IgA in rectal secretions. N(imm) was superior to V-FP(imm) for producing rectal CTB-specific IgA, but the greatest amounts of CTB-specific IgA and LPS specific IgA, IgG, and IgM Ab were found in rectal secretions of R(imm) women. These data suggest that in women, N(imm) alone could induce specific Ab in serum, the genital tract, and rectum. However, induction of genital tract and rectal Ab responses of the magnitude generated by local V-FP(imm) or R(imm) will likely require administration of comparably high nasal vaccine dosages. PMID- 12077290 TI - Generation of CTL recognizing an HLA-A*0201-restricted epitope shared by MAGE-A1, -A2, -A3, -A4, -A6, -A10, and -A12 tumor antigens: implication in a broad spectrum tumor immunotherapy. AB - MAGE-A1, -A2, -A3, -A4, -A6, -A10, and -A12 are expressed in a significant proportion of primary and metastatic tumors of various histological types and are targets of tumor Ag-specific CTL. Individual MAGE-A expression varies from one tumor type to the other but, overall, the large majority of tumors expresses at least one MAGE-A Ag. Therefore, targeting epitopes shared by all MAGE-A Ags would be of interest in immunotherapy against a broad spectrum of cancers. In the present study, we describe a heteroclitic MAGE-A peptide (p248V9) that induces CTL in vivo in HLA-A*0201 transgenic HHD mice and in vitro in healthy donors. These CTL are able to recognize two low HLA-A*0201 affinity peptides differing at their C-terminal position and derived from MAGE-A2, -A3, -A4, -A6, -A10, and -A12 (p248G9) and MAGE-A1 (p248D9). Interestingly, p248V9-specific CTL respond to endogenous MAGE-A1, -A2, -A3, -A4, -A6, -A10, and -A12 in an HLA-A*0201 restricted manner and recognize human HLA-A*0201(+)MAGE-A(+) tumor cells of various histological origin. Therefore, this heteroclitic peptide may be considered as a potent candidate for a broad-spectrum tumor vaccination. PMID- 12077291 TI - Aberrant production of IL-12 by macrophages from several autoimmune-prone mouse strains is characterized by intrinsic and unique patterns of NF-kappa B expression and binding to the IL-12 p40 promoter. AB - Intrinsic defects in macrophage (M(phi)) cytokine production characterize many autoimmune-prone mouse strains. Aberrant levels of IL-12, for example, are produced by M(phi) isolated from young mice prone to lupus (MRL and NZB/W) and diabetes (nonobese diabetic (NOD)) well before the appearance of disease signs. Evaluation of the possible mechanism(s) underlying the abnormal regulation of IL 12 in these strains revealed novel patterns of Rel family protein binding to the unique p40 NF-kappaB site in the IL-12 p40 promoter, whereas binding patterns to Ets and CCAAT enhancer binding protein/beta sites were normal. In particular, the heightened production of IL-12 by NOD M(phi) is associated with elevated levels of the trans-activating p50/c-Rel (p65) complex compared with the nonfunctional p50/p50 dimer. Conversely, the dramatically impaired production of IL-12 by both NZB/W and MRL/+ M(phi) is associated with a predominance of p50/p50 and reduced p50/c-Rel(p65) binding. Mechanistically, the unique pattern seen in the lupus strains reflects elevated p50 and reduced c-Rel nuclear protein levels. In NOD extracts, the level of c-Rel is elevated compared with that in lupus strains, but not when compared with that in normal A/J. However, the extent of c-Rel tyrosine phosphorylation noted in NOD extracts is more than double that seen in any other strain. Levels of p65 were similar in all strains tested. These findings reveal that a common mechanism, involving dysregulation of c-Rel and p50, may be used to determine the aberrant IL-12 levels that have the potential to predispose specific mouse strains to systemic or organ-specific autoimmunity. PMID- 12077292 TI - Induction of autoantibody production is limited in nonautoimmune mice. AB - Many individuals develop a single or a few brief episodes of autoimmunity from which they recover. Mechanisms that quell pathologic autoimmunity following such a breakdown of self-tolerance are not clearly understood. In this study, we show that in nonautoimmune mice, dsDNA-specific autoreactive B cells exist but remain inactive. This state of inactivation in dsDNA-specific B cells could be disrupted by autoreactive Th cells; in this case T cells that react with peptides from the V(H) region of anti-DNA Abs (hereafter called anti-V(H) T cells). Immunization with anti-DNA mAb, its gamma-chain or peptides derived from its V(H) region induced anti-V(H) Th cells, IgG anti-dsDNA Ab, and proteinuria. The breakdown of B cell tolerance in nonautoimmune mice, however, was short-lived: anti-DNA Ab and nephritis subsided despite subsequent immunizations. The recovery from autoimmunity temporally correlated with the appearance of T cells that inhibited anti-DNA Ab production. Such inhibitory T cells secreted TGFbeta; the inhibition of anti-DNA Ab production by these cells was partly abolished by anti-TGFbeta Ab. Even without immunization, nonautoimmune mice possess T cells that can inhibit autoantibody production. Thus, inhibitory T cells in nonautoimmune mice may normally inhibit T-dependent activation of autoreactive B cells and/or reverse such activation following stimulation by Th cells. The induction of such inhibitory T cells may play a role in protecting nonautoimmune mice from developing chronic autoimmunity. PMID- 12077293 TI - Efficient isolation of novel human monoclonal antibodies with neutralizing activity against HIV-1 from transgenic mice expressing human Ig loci. AB - Despite considerable interest in the isolation of mAbs with potent neutralization activity against primary HIV-1 isolates, both for identifying useful targets for vaccine development and for the development of therapeutically useful reagents against HIV-1 infection, a relatively limited number of such reagents have been isolated to date. Human mAbs (hu-mAbs) are preferable to rodent mAbs for treatment of humans, but isolation of hu-mAbs from HIV-infected subjects by standard methods of EBV transformation of B cells or phage display of Ig libraries is inefficient and limited by the inability to control or define the original immunogen. An alternative approach for the isolation of hu-mAbs has been provided by the development of transgenic mice that produce fully hu-mAbs. In this report, we show that immunizing the XenoMouse G2 strain with native recombinant gp120 derived from HIV(SF162) resulted in robust humoral Ab responses against gp120 and allowed the efficient isolation of hybridomas producing specific hu-mAbs directed against multiple regions and epitopes of gp120. hu-mAbs possessing strong neutralizing activity against the autologous HIV(SF162) strain were obtained. The epitopes recognized were located in three previously described neutralization domains, the V2-, V3- and CD4-binding domains, and in a novel neutralization domain, the highly variable C-terminal region of the V1 loop. This is the first report of neutralizing mAbs directed at targets in the V1 region. Furthermore, the V2 and V3 epitopes recognized by neutralizing hu-mAbs were distinct from those of previously described human and rodent mAbs and included an epitope requiring a full length V3 loop peptide for effective presentation. These results further our understanding of neutralization targets for primary, R5 HIV-1 viruses and demonstrate the utility of the XenoMouse system for identifying new and interesting epitopes on HIV-1. PMID- 12077294 TI - Zinc finger as distance determinant in the flexible linker of intron endonuclease I-TevI. AB - I-TevI, the phage T4 td intron-encoded endonuclease, recognizes a lengthy DNA target and initiates intron mobility by introducing a double-strand break in the homing site. The enzyme uses both sequence and distance determinants to cleave the DNA 23-25 bp upstream of the intron insertion site. I-TevI consists of an N terminal catalytic domain and a C-terminal DNA-binding domain separated by a long, flexible linker. The DNA-binding domain consists of three subdomains: a zinc finger, a minor-groove binding alpha-helix, and a helix-turn-helix. In this study, a mutational analysis was undertaken to assess the roles of these subdomains in substrate binding and cleavage. Surprisingly, the zinc finger is not required for DNA binding or catalysis. Rather, the zinc finger is a component of the linker and directs the catalytic domain to cleave the homing site at a fixed distance from the intron insertion site. When the cleavage site (CS) is shifted outside a given range, wild-type I-TevI defaults to the fixed distance, whereas zinc-finger mutants have lost the distance determinant and search out the displaced cleavage sequences. Although counterintuitive, a protein containing a 19-aa deletion of the zinc finger can extend further than can wild-type I-TevI to cleave a distant CS sequence, and a Cys-to-Ala mutant of the ligands for zinc, nominally a longer protein, can retract to cleave at a closer CS sequence. Models are presented for the novel function of the zinc finger, as a molecular constraint, whereby intramolecular protein-protein interactions position the catalytic domain by "catalytic clamp" and/or "linker-organizer" mechanisms. PMID- 12077295 TI - Inhibition of mitochondrial respiration by endogenous nitric oxide: a critical step in Fas signaling. AB - We have found that activation of human adult T cell leukemia (Jurkat) cells with anti-Fas Ab leads, in a concentration-dependent manner, to an early burst of production of nitric oxide (NO), which inhibits cell respiration. This results in mitochondrial hyperpolarization, dependent on the hydrolysis of glycolytic ATP by the F1F(o)-ATPase acting in reverse mode. During this early phase of activation, there is a transient release of superoxide anion. All these processes can be prevented by an inhibitor of NO synthase. Approximately 2 h after stimulation with anti-Fas Ab, a distinct second phase can be detected. This comprises a concentration-dependent collapse in mitochondrial membrane potential, a second wave of free radical production, and activation of caspase-8 leading to apoptosis. This second phase is abolished by an inhibitor of caspase activation. In contrast, inhibition of NO synthesis leads to an enhancement and acceleration of these latter processes, suggesting that the early NO-dependent phase represents a protective mechanism. The significance of the two phases in relation to cell survival and death remains to be studied. PMID- 12077296 TI - The etiology of white pox, a lethal disease of the Caribbean elkhorn coral, Acropora palmata. AB - Populations of the shallow-water Caribbean elkhorn coral, Acropora palmata, are being decimated by white pox disease, with losses of living cover in the Florida Keys typically in excess of 70%. The rate of tissue loss is rapid, averaging 2.5 cm2 x day(-1), and is greatest during periods of seasonally elevated temperature. In Florida, the spread of white pox fits the contagion model, with nearest neighbors most susceptible to infection. In this report, we identify a common fecal enterobacterium, Serratia marcescens, as the causal agent of white pox. This is the first time, to our knowledge, that a bacterial species associated with the human gut has been shown to be a marine invertebrate pathogen. PMID- 12077297 TI - Genome expression analysis of Anopheles gambiae: responses to injury, bacterial challenge, and malaria infection. AB - The complex gene expression responses of Anopheles gambiae to microbial and malaria challenges, injury, and oxidative stress (in the mosquito and/or a cultured cell line) were surveyed by using cDNA microarrays constructed from an EST-clone collection. The expression profiles were broadly subdivided into induced and down-regulated gene clusters. Gram+ and Gram- bacteria and microbial elicitors up-regulated a diverse set of genes, many belonging to the immunity class, and the response to malaria partially overlapped with this response. Oxidative stress activated a distinctive set of genes, mainly implicated in oxidoreductive processes. Injury up- and down-regulated gene clusters also were distinctive, prominently implicating glycolysis-related genes and citric acid cycle/oxidative phosphorylation/redox-mitochondrial functions, respectively. Cross-comparison of in vivo and in vitro responses indicated the existence of tightly coregulated gene groups that may correspond to gene pathways. PMID- 12077298 TI - Rapid natural scene categorization in the near absence of attention. AB - What can we see when we do not pay attention? It is well known that we can be "blind" even to major aspects of natural scenes when we attend elsewhere. The only tasks that do not need attention appear to be carried out in the early stages of the visual system. Contrary to this common belief, we report that subjects can rapidly detect animals or vehicles in briefly presented novel natural scenes while simultaneously performing another attentionally demanding task. By comparison, they are unable to discriminate large T's from L's, or bisected two-color disks from their mirror images under the same conditions. We conclude that some visual tasks associated with "high-level" cortical areas may proceed in the near absence of attention. PMID- 12077299 TI - Interaction of G alpha(12) with G alpha(13) and G alpha(q) signaling pathways. AB - The G(12) subfamily of heterotrimeric G-proteins consists of two members, G(12) and G(13). Gene-targeting studies have revealed a role for G(13) in blood vessel development. Mice lacking the alpha subunit of G(13) die around embryonic day 10 as the result of an angiogenic defect. On the other hand, the physiological role of G(12) is still unclear. To address this issue, we generated G alpha(12) deficient mice. In contrast to the G alpha(13)-deficient mice, G alpha(12) deficient mice are viable, fertile, and do not show apparent abnormalities. However, G alpha(12) does not seem to be entirely redundant, because in the offspring generated from G alpha(12)+/- G alpha(13)+/- intercrosses, at least one intact G alpha(12) allele is required for the survival of animals with only one G alpha(13) allele. In addition, G alpha(12) and G alpha(13) showed a difference in mediating cell migratory response to lysophosphatidic acid in embryonic fibroblast cells. Furthermore, mice lacking both G alpha(12) and G alpha(q) die in utero at about embryonic day 13. These data indicate that the G alpha(12) mediated signaling pathway functionally interacts not only with the G alpha(13)- but also with the G alpha(q/11)-mediated signaling systems. PMID- 12077300 TI - Transformation of follicular lymphoma to diffuse large-cell lymphoma: alternative patterns with increased or decreased expression of c-myc and its regulated genes. AB - The natural history of follicular lymphoma (FL) is frequently characterized by transformation to a more aggressive diffuse large B cell lymphoma (DLBCL). We compared the gene-expression profiles between transformed DLBCL and their antecedent FL. No genes were observed to increase or decrease their expression in all of the cases of histological transformation. However, two different gene expression profiles associated with the transformation process were defined, one in which c-myc and genes regulated by c-myc showed increased expression and one in which these same genes showed decreased expression. Further, there was a striking difference in gene-expression profiles between transformed DLBCL and de novo DLBCL, because the gene-expression profile of transformed DLBCL was more similar to their antecedent FL than to de novo DLBCL. This study demonstrates that transformation from FL to DLBCL can occur by alternative pathways and that transformed DLBCL and de novo DLBCL have very different gene-expression profiles that may underlie the different clinical behaviors of these two types of morphologically similar lymphomas. PMID- 12077301 TI - Dome formation in cell cultures as expression of an early stage of lactogenic differentiation of the mammary gland. AB - The study of the development of the mammary gland at the molecular level in the animal is difficult because of the complex tissue organization of the gland. We have previously developed an in vitro system for genetic analysis of mammary cell differentiation, based on the cell line LA7 clonally derived from a rat mammary adenocarcinoma. This cell line, after induction with DMSO, differentiates forming structures called domes. This process is under strict gene regulation, and we have previously identified several of the genes involved. In the present paper, we have defined the meaning of dome formation in relation to mammary development, by showing that treatment of LA7 cells with the lactogenic hormones hydrocortisone and prolactin induces dome formation; in the animal, these hormones precede and accompany milk production. Moreover, dome formation is accompanied by expression within the cells of the milk protein genes WDMN1 and beta-casein, which are differentiation markers for the gland during pregnancy and lactation. We also show that two proteins, highly expressed in the mammary gland during lactation, HSP90-beta and annexin I, are strongly expressed in DMSO induced LA7 cells. Both proteins are essential in the formation of domes because when their synthesis is blocked by antisense RNA oligonucleotides, dome formation is abolished. Thus our in vitro system is a model for lobulo-alveolar development, and the genes identified in the pathway of dome formation are likely to be involved in the early differentiation steps occurring in the rat mammary gland during pregnancy and lactation. PMID- 12077303 TI - Semidwarf (sd-1), "green revolution" rice, contains a defective gibberellin 20 oxidase gene. AB - The introduction of semidwarf rice (Oryza sativa L.) led to record yield increases throughout Asia in the 1960s. The major semidwarfing allele, sd-1, is still extensively used in modern rice cultivars. The phenotype of sd-1 is consistent with dwarfism that results from a deficiency in gibberellin (GA) plant growth hormones. We propose that the semidwarf (sd-1) phenotype is the result of a deficiency of active GAs in the elongating stem arising from a defective 20 oxidase GA biosynthetic enzyme. Sequence data from the rice genome was combined with previous mapping studies to locate a putative GA 20-oxidase gene (Os20ox2) at the predicted map location of sd-1 on chromosome 1. Two independent sd-1 alleles contained alterations within Os20ox2: a deletion of 280 bp within the coding region of Os20ox2 was predicted to encode a nonfunctional protein in an indica type semidwarf (Doongara), whereas a substitution in an amino acid residue (Leu-266) that is highly conserved among dioxygenases could explain loss of function of Os20ox2 in a japonica semidwarf (Calrose76). The quantification of GAs in elongating stems by GC-MS showed that the initial substrate of GA 20 oxidase activity (GA53) accumulated, whereas the content of the major product (GA20) and of bioactive GA1 was lower in semidwarf compared with tall lines. We propose that the Os20ox2 gene corresponds to the sd-1 locus. PMID- 12077302 TI - A possible evolutionary origin for the Mn4 cluster of the photosynthetic water oxidation complex from natural MnO2 precipitates in the early ocean. AB - The photosynthetic water oxidation complex consists of a cluster of four Mn atoms bridged by O atoms, associated with Ca2+ and Cl-, and incorporated into protein. The structure is similar in higher plants and algae, as well as in cyanobacteria of more ancient lineage, dating back more than 2.5 billion years ago on Earth. It has been proposed that the proto-enzyme derived from a component of a natural early marine manganese precipitate that contained a CaMn4O9 cluster. A variety of MnO2 minerals are found in nature. Three major classes are spinels, sheet-like layered structures, and three-dimensional networks that contain parallel tunnels. These relatively open structures readily incorporate cations (Na+, Li+, Mg2+, Ca2+, Ba2+, H+, and even Mn2+) and water. The minerals have different ratios of Mn(III) and Mn(IV) octahedrally coordinated to oxygens. Using x-ray spectroscopy we compare the chemical structures of Mn in the minerals with what is known about the arrangement in the water oxidation complex to define the parameters of a structural model for the photosynthetic catalytic site. This comparison provides for the structural model a set of candidate Mn(4) clusters-some previously proposed and considered and others entirely novel. PMID- 12077304 TI - Employing Escherichia coli to functionally express, purify, and characterize a human transporter. AB - Large-scale purification of recombinant human membrane proteins represents a rate limiting step toward the understanding of their role in health and disease. There are only four mammalian membrane proteins of known structure, and these were isolated from natural sources (see http://www.mpibp frankfurt.mpg.de/michel/public/memprotstruct.html). In addition, genetic diseases of membrane proteins are frequently caused by trafficking defects, and it is enigmatic whether these mutants are functional. Here, we report the employment of Escherichia coli for the functional expression, purification, and reconstitution of a human membrane protein, the human Na+/glucose cotransporter (hSGLT1). The use of an E. coli mutant defective in the outer membrane protease OmpT, incubation temperatures below 20 degrees C, and transcriptional regulation from the lac promoter/operator are crucial to reduce proteolytic degradation. Purification of a recombinant hSGLT1 through affinity chromatography yields about 1 mg of purified recombinant hSGLT1 per 3 liters of cultured bacterial cells. Kinetic analysis of hSGLT1 in proteoliposomes reveals that a purified recombinant transporter, which is missorted in eukaryotic cells, retains full catalytic activity. These results indicate the power of bacteria to manufacture and isolate human membrane proteins implicated in genetic diseases. PMID- 12077305 TI - On the evolution of cells. AB - A theory for the evolution of cellular organization is presented. The model is based on the (data supported) conjecture that the dynamic of horizontal gene transfer (HGT) is primarily determined by the organization of the recipient cell. Aboriginal cell designs are taken to be simple and loosely organized enough that all cellular componentry can be altered and/or displaced through HGT, making HGT the principal driving force in early cellular evolution. Primitive cells did not carry a stable organismal genealogical trace. Primitive cellular evolution is basically communal. The high level of novelty required to evolve cell designs is a product of communal invention, of the universal HGT field, not intralineage variation. It is the community as a whole, the ecosystem, which evolves. The individual cell designs that evolved in this way are nevertheless fundamentally distinct, because the initial conditions in each case are somewhat different. As a cell design becomes more complex and interconnected a critical point is reached where a more integrated cellular organization emerges, and vertically generated novelty can and does assume greater importance. This critical point is called the "Darwinian Threshold" for the reasons given. PMID- 12077306 TI - The p53MH algorithm and its application in detecting p53-responsive genes. AB - A computer algorithm, p53MH, was developed, which identifies putative p53 transcription factor DNA-binding sites on a genomewide scale with high power and versatility. With the sequences from the human and mouse genomes, putative p53 DNA-binding elements were identified in a scan of 2,583 human genes and 1,713 mouse orthologs based on the experimental data of el-Deiry et al. [el-Deiry, W. S., Kern, S. E., Pietenpol, J. A., Kinzler, K. W. & Vogelstein, B. (1992) Nat. Genet. 1, 45-49] and Funk et al. [Funk, W. D., Pak, D. T., Karas, R. H., Wright, W. E. & Shay, J. W. (1992) Mol. Cell. Biol. 12, 2866-2871] (http://linkage.rockefeller.edu/p53). The p53 DNA-binding motif consists of a 10 bp palindrome and most commonly a second related palindrome linked by a spacer region. By scanning from the 5' to 3' end of each gene with an additional 10-kb nucleotide sequence appended at each end (most regulatory DNA elements characterized in the literature are in these regions), p53MH computes the binding likelihood for each site under a discrete discriminant model and then outputs ordered scores, corresponding site positions, sequences, and related information. About 300 genes receiving scores greater than a theoretical cut-off value were identified as potential p53 targets. Semiquantitative reverse transcription-PCR experiments were performed in 2 cell lines on 16 genes that were previously unknown regarding their functional relationship to p53 and were found to have high scores in either proximal promoter or possible distal enhancer regions. Ten (approximately 63%) of these genes responded to the presence of p53. PMID- 12077307 TI - Emergence of multiple genotypes of H5N1 avian influenza viruses in Hong Kong SAR. AB - Although A/Hong Kong/156/97 (H5N1/97)-like viruses associated with the "bird flu" incident in Hong Kong SAR have not been detected since the slaughter of poultry in 1997, its putative precursors continue to persist in the region. One of these, Goose/Guangdong/1/96 (H5N1 Gs/Gd)-like viruses, reassorted with other avian viruses to generate multiple genotypes of H5N1 viruses that crossed to chickens and other terrestrial poultry from its reservoir in geese. Whereas none of these recent reassortants had acquired the gene constellation of H5N1/97, these events provide insight into how such a virus may have been generated. The recent H5N1 reassortants readily infect and kill chicken and quail after experimental infection, and some were associated with significant mortality of chickens within the poultry retail markets in Hong Kong. Some genotypes are lethal for mice after intra-nasal inoculation and spread to the brain. On this occasion, the early detection of H5N1 viruses in the retail, live poultry markets led to preemptive intervention before the occurrence of human disease, but these newly emerging, highly pathogenic H5N1 viruses provide cause for pandemic concern. PMID- 12077309 TI - Activation of Cdc2/cyclin B and inhibition of centrosome amplification in cells depleted of Plk1 by siRNA. AB - The events of the cell cycle, the stages at which the cell proliferates and divides, are facilitated and controlled by multiple signaling pathways. Among the many regulatory enzymes that contribute to these processes is the polo-like kinase (Plk). Plks have been reported to mediate multiple mitotic processes, including bipolar spindle formation, activation of Cdc25C, actin ring formation, centrosome maturation, and activation of the anaphase-promoting complex. To investigate its functions in mammalian cells further, we used the recently developed small interfering RNA technique specifically to deplete Plk1 in cultured cells. We find that Plk1 depletion results in elevated Cdc2 protein kinase activity and thus attenuates cell-cycle progression. About 45% of cells treated with Plk1 small interfering RNA show the formation of a dumbbell-like DNA organization, suggesting that sister chromatids are not completely separated. About 15% of these cells do complete anaphase but do not complete cytokinesis. Finally, Plk1 depletion significantly reduces centrosome amplification in hydroxyurea-treated U2OS cells. These data provide direct evidence that Plk is required for multiple mitotic processes in mammalian cells and their significance is discussed. PMID- 12077310 TI - Transcriptional collision between convergent genes in budding yeast. AB - Transcriptional interference between genes and the regulatory elements of simple eukaryotes such as Saccharomyces cerevisiae is an unavoidable consequence of their compressed genetic arrangement. We have shown previously that with the tandem arranged genes GAL10 and GAL7, inefficient transcriptional termination of the upstream gene inhibits initiation of transcription on the downstream gene. We now show that transcriptional interference can occur also with S. cerevisiae RNA polymerase II genes arranged convergently. We demonstrate that when the GAL10 and GAL7 genes are rearranged in a convergent orientation, transcriptional initiation occurs at full levels. However, as soon as the two transcripts begin to overlap, elongation is restricted, resulting in a severe reduction in steady-state mRNA accumulation. This effect is observed only in cis arrangement, arguing against RNA-interference effects acting on the potential generation of antisense transcripts. These data reinforce the necessity of separating adjacent RNA polymerase II transcription units by efficient termination signals. PMID- 12077311 TI - Identification of a gene encoding MGAT1, a monoacylglycerol acyltransferase. AB - Acyl-CoA:monoacylglycerol acyltransferase (MGAT) catalyzes the synthesis of diacylglycerol, the precursor of physiologically important lipids such as triacylglycerol and phospholipids. In the intestine, MGAT plays a major role in the absorption of dietary fat because resynthesis of triacylglycerol is required for the assembly of lipoproteins that transport absorbed fat to other tissues. MGAT activity has also been reported in mammalian liver and white adipose tissue. However, MGAT has never been purified to homogeneity from mammalian tissues, and its gene has not been cloned. We identified a gene that encodes an MGAT (MGAT1) in mice. This gene has sequence homology with members of a recently identified diacylglycerol acyltransferase gene family. Expression of the MGAT1 cDNA in insect cells markedly increased MGAT activity in cell membranes. In addition, MGAT activity was proportional to the level of MGAT1 protein expressed, and the amount of diacylglycerol produced depended on the concentration of either of its substrates, oleoyl-CoA or monooleoylglycerol. In mice, MGAT1 expression and MGAT activity were detected in the stomach, kidney, white and brown adipose tissue, and liver. However, MGAT1 was not expressed in the small intestine, implying the existence of a second MGAT gene. The identification of the MGAT1 gene should greatly facilitate research on the identification of the intestinal MGAT gene and on the function of MGAT enzymes in mammalian glycerolipid metabolism. PMID- 12077312 TI - Transcriptional response of Saccharomyces cerevisiae to DNA-damaging agents does not identify the genes that protect against these agents. AB - The recent completion of the deletion of all of the nonessential genes in budding yeast has provided a powerful new way of determining those genes that affect the sensitivity of this organism to cytotoxic agents. We have used this system to test the hypothesis that genes whose transcription is increased after DNA damage are important for the survival to that damage. We used a pool of 4,627 diploid strains each with homozygous deletion of a nonessential gene to identify those genes that are important for the survival of yeast to four DNA-damaging agents: ionizing radiation, UV radiation, and exposure to cisplatin or to hydrogen peroxide. In addition we measured the transcriptional response of the wild-type parental strain to the same DNA-damaging agents. We found no relationship between the genes necessary for survival to the DNA-damaging agents and those genes whose transcription is increased after exposure. These data show that few, if any, of the genes involved in repairing the DNA lesions produced in this study, including double-strand breaks, pyrimidine dimers, single-strand breaks, base damage, and DNA cross-links, are induced in response to toxic doses of the agents that produce these lesions. This finding suggests that the enzymes necessary for the repair of these lesions are at sufficient levels within the cell. The data also suggest that the nature of the lesions produced by DNA-damaging agents cannot easily be deduced from gene expression profiling. PMID- 12077313 TI - Quantitative evolutionary design of nutrient processing: glucose. AB - Quantitative evolutionary design involves the numerical relationships, evolved through natural selection, of biological capacities to each other and to natural loads. Here we study the relation of nutrient-processing capacities of the intestine and of organs beyond it (such as liver and kidneys) to each other and to natural loads of nutrients normally consumed. To control experimentally the rate of nutrient delivery to organs beyond the intestine, we administered nutrients directly into the veins of rats by the method of total parenteral nutrition (TPN). Control rats consuming the TPN solution by mouth ingested glucose at 42 mmol/day and processed it completely, as gauged by negligible appearance of glucose in urine and feces. Experimental rats receiving TPN were able to process infused glucose completely at rates up to 92 mmol/day. At higher infusion rates, they were unable to process further glucose, as gauged by rises in serum and urinary glucose levels and serum osmolality. At the highest infusion rates, they exhibited diuresis, dehydration, and both decreased weight gain and survival. These symptoms closely resemble the human diabetic condition known as nonketotic hypertonicity. Thus, a rat's body has a safety factor of 2.2 (=92/42) for glucose processing: it can process glucose at a rate 2.2 times its voluntary intake. This safety factor represents apparent excess capacity that may have evolved to process other nutrients converted into glucose, to minimize the risk of loads swamping capacities, to handle suddenly increased nutrient requirements, or to effect rapid mobilization of glucose. PMID- 12077314 TI - Microbial subversion of the immune response. PMID- 12077315 TI - Adaptive immunity cooperates with liposomal all-trans-retinoic acid (ATRA) to facilitate long-term molecular remissions in mice with acute promyelocytic leukemia. AB - We previously developed a murine model of acute promyelocytic leukemia (APL) by using human cathepsin G gene regulatory elements to direct the expression of promyelocytic leukemia (PML)/retinoic acid receptor alpha (RAR alpha) and RAR alpha/PML fusion cDNAs to the early myeloid compartment of transgenic mice. To study the efficacy of noncytotoxic therapy in this animal model, cohorts of naive immunocompetent mice were inoculated with primary murine APL cells from a frozen tumor bank. Arsenic trioxide and liposomally encapsulated all-trans-retinoic acid (Lipo ATRA), alone or in combination, were administered for 21 days by i.p. injection using doses that yielded plasma levels similar to those observed in human APL patients treated with these agents. Lipo ATRA was highly effective in inducing durable molecular remissions in immunocompetent mice [C57BL/6 x C3H F(1) (B6C3HF1)]; arsenic therapy was much less effective, and did not clearly synergize with Lipo ATRA to increase the remission rate in immunocompetent mice. The survival of Lipo ATRA-treated severe combined immunodeficient (SCID) animals (lacking functional T and B cells) was inferior to that of immunocompetent B6C3HF1 recipients (40% vs. 88% survival at 1 y, P < 0.001). These data suggest that adaptive immunity cooperates with pharmacologic therapy to induce or maintain remissions in murine APL. It also implies that immunosuppressive anti leukemia therapies could paradoxically blunt effective anti-leukemia immune responses that are important for clearing small numbers of residual tumor cells after chemotherapy-mediated cytoreduction. PMID- 12077316 TI - Pheromone induction promotes Ste11 degradation through a MAPK feedback and ubiquitin-dependent mechanism. AB - Ste11 is the mitogen-activated protein kinase (MAPK) kinase kinase in the MAPK cascades that mediate mating, high osmolarity glycerol, and filamentous growth responses in Saccharomyces cerevisiae. We show stimulation of the mating pathway by pheromone promotes an accelerated turnover of Ste11 through a MAPK feedback and ubiquitin-dependent mechanism. This degradation is pathway specific, because Ste11 is stable during activation of the high osmolarity glycerol pathway. Because the steady-state amount of Ste11 does not change significantly during pheromone induction, we infer that maintenance of MAPK activation involves repeated cycles in which naive Ste11 is activated and then targeted for degradation. This model predicts that elimination of active Ste11 would rapidly curtail MAPK activation upon attenuation of the upstream signal. This prediction is confirmed by the finding that blocking ubiquitin-dependent Ste11 degradation during pheromone induction abolishes the characteristic attenuation profile for MAPK activation. PMID- 12077317 TI - A ribozyme that ligates RNA to protein. AB - We have used a combination of in vitro selection and rational design to generate ribozymes that form a stable phosphoamide bond between the 5' terminus of an RNA and a specific polypeptide. This reaction differs from that of previously identified ribozymes, although the product is analogous to the enzyme-nucleotidyl intermediates isolated during the reactions of certain proteinaceous enzymes, such as guanyltransferase, DNA ligase, and RNA ligase. Comparative sequence analysis of the isolated ribozymes revealed that they share a compact secondary structure containing six stems arranged in a four-helix junction and branched pseudoknot. An optimized version of the ribozyme reacts with substrate-fusion proteins, allowing it to be used to attach RNA tags to proteins both in vitro and within bacterial cells, suggesting a simple way to tag a specific protein with amplifiable information. PMID- 12077318 TI - The barley stem rust-resistance gene Rpg1 is a novel disease-resistance gene with homology to receptor kinases. AB - Stem rust caused by Puccinia graminis f. sp. tritici was among the most devastating diseases of barley in the northern Great Plains of the U.S. and Canada before the deployment of the stem rust-resistance gene Rpg1 in 1942. Since then, Rpg1 has provided durable protection against stem rust losses in widely grown barley cultivars (cvs.). Extensive efforts to clone Rpg1 by synteny with rice provided excellent flanking markers but failed to yield the gene because it does not seem to exist in rice. Here we report the map-based cloning and characterization of Rpg1. A high-resolution genetic map constructed with 8,518 gametes and a 330-kb bacterial artificial chromosome contig physical map positioned the gene between two crossovers approximately 0.21 centimorgan and 110 kb apart. The region including Rpg1 was searched for potential candidate genes by sequencing low-copy probes. Two receptor kinase-like genes were identified. The candidate gene alleles were sequenced from resistant and susceptible cvs. Only one of the candidate genes showed a pattern of apparently functional gene structure in the resistant cvs. and defective gene structure in the susceptible cvs. identifying it as the Rpg1 gene. Rpg1 encodes a receptor kinase-like protein with two tandem protein kinase domains, a novel structure for a plant disease resistance gene. Thus, it may represent a new class of plant resistance genes. PMID- 12077319 TI - Overexpression, purification, and site-directed spin labeling of the Nramp metal transporter from Mycobacterium leprae. AB - It has long been recognized that the pathogenicity of a broad range of intracellular parasites depends on the availability of transition metal ions, especially iron. Nramp1 (natural resistance-associated macrophage protein 1), a proton-coupled divalent metal ion transporter, has been identified as a controlling factor in the resistance or susceptibility to infection with a diverse range of intracellular pathogens such as Toxoplasma, Salmonella, Mycobacterium, and Leishmania. The role of divalent metal ion transport is even more compelling given the existence of Nramp homologs in several intracellular parasites, such as mycobacteria. We have confirmed the functional homology of the Nramp homologue from Mycobacterium leprae by using a yeast complementation assay for divalent cation uptake. To facilitate a concerted biochemical and structural analysis of this important class of transporters, the M. leprae Nramp was expressed in Escherichia coli. Dual affinity tags were engineered at the N and C termini to allow for isolation of full-length protein at >95% purity. Site directed spin labeling of Cys-299 reveals a flexible hinge-like domain. A weak dipolar interaction is detected between the nitroxide and paramagnetic transition ions, indicating this position is approximately 19 A from the nearest high affinity binding site. PMID- 12077320 TI - Kinetic analysis of estrogen receptor/ligand interactions. AB - Surface plasmon resonance biosensor technology was used to directly measure the binding interactions of small molecules to the ligand-binding domain of human estrogen receptor. In a screening mode, specific ligands of the receptor were easily discerned from nonligands. In a high-resolution mode, the association and dissociation phase binding responses were shown to be reproducible and could be fit globally to a simple interaction model to extract reaction rate constants. On average, antagonist ligands (such as tamoxifen and nafoxidine) were observed to bind to the receptor with association rates that were 500-fold slower than agonists (such as estriol and beta-estradiol). This finding is consistent with these antagonists binding to an altered conformation of the receptor. The biosensor assay also could identify subtle differences in how the same ligand interacted with two different isoforms of the receptor (alpha and beta). The biosensor's ability to determine kinetic rate constants for small molecule/protein interactions provides unique opportunities to understand the mechanisms associated with complex formation as well as new information to drive the optimization of drug candidates. PMID- 12077321 TI - Association of a lysine-232/alanine polymorphism in a bovine gene encoding acyl CoA:diacylglycerol acyltransferase (DGAT1) with variation at a quantitative trait locus for milk fat content. AB - DGAT1 encodes diacylglycerol O-acyltransferase (EC ), a microsomal enzyme that catalyzes the final step of triglyceride synthesis. It became a functional candidate gene for lactation traits after studies indicated that mice lacking both copies of DGAT1 are completely devoid of milk secretion, most likely because of deficient triglyceride synthesis in the mammary gland. Our mapping studies placed DGAT1 close to the region of a quantitative trait locus (QTL) on bovine chromosome 14 for variation in fat content of milk. Sequencing of DGAT1 from pooled DNA revealed significant frequency shifts at several variable positions between groups of animals with high and low breeding values for milk fat content in different breeds (Holstein-Friesian, Fleckvieh, and Braunvieh). Among the variants was a nonconservative substitution of lysine by alanine (K232A), with the lysine-encoding allele being associated with higher milk fat content. Haplotype analysis indicated the lysine variant to be ancestral. Two animals that were typed heterozygous (Qq) at the QTL based on marker-assisted QTL-genotyping were heterozygous for the K232A substitution, whereas 14 animals that are most likely qq at the QTL were homozygous for the alanine-encoding allele. An independent association study in Fleckvieh animals confirmed the positive effect of the lysine variant on milk fat content. We consider the nonconservative K232A substitution to be directly responsible for the QTL variation, although our genetic studies cannot provide formal proof. PMID- 12077322 TI - CD28 plays a critical role in the segregation of PKC theta within the immunologic synapse. AB - The signaling pathways that lead to the localization of cellular protein to the area of interaction between T cell and antigen-presenting cell and the mechanism by which these molecules are further sorted to the peripheral supramolecular activation cluster or central supramolecular activation cluster regions of the immunologic synapse are poorly understood. In this study, we investigated the functional involvement of CD28 costimulation in the T cell receptor (TCR) mediated immunologic synapse formation with respect to protein kinase C (PKC)theta; localization. We showed that CD3 crosslinking alone was sufficient to induce PKC theta; capping in naive CD4(+) T cells. Studies with pharmacologic inhibitors and knockout mice showed that the TCR-derived signaling that drives PKC theta; membrane translocation requires the Src family kinase, Lck, but not Fyn. In addition, a time course study of the persistence of T cell molecules to the immunologic synapse indicated that PKC theta;, unlike TCR, persisted in the synapse for at least 4 h, a time that is sufficient for commitment of a T cell to cell division. Finally, by using TCR-transgenic T cells from either wild-type or CD28-deficient mice, we showed that CD28 expression was required for the formation of the mature immunologic synapse, because antigen stimulation of CD28( ) T cells led to a diffuse pattern of localization of PKC theta; and lymphocyte function-associated antigen-1 in the immunologic synapse, in contrast to the central supramolecular activation cluster localization of PKC theta; in CD28(+) T cells. PMID- 12077323 TI - GATA-factor dependence of the multitype zinc-finger protein FOG-1 for its essential role in megakaryopoiesis. AB - The function of GATA transcription factors in diverse developmental contexts depends in part on physical interaction with cofactors of the Friend of GATA (FOG) family. However, previous studies indicate that FOG-1 may play a GATA-1 independent role in early megakaryopoiesis, suggesting that FOG proteins might act in a GATA factor-independent manner. Here, we have generated mouse knock-in (KI) mutants harboring a critical valine-to-glycine substitution in the amino terminal zinc fingers of GATA-1 and GATA-2 to ablate FOG interaction. In contrast to male GATA-1(KI) (GATA-1 is located on the X-chromosome) or GATA-2(KI/KI) mice, compound GATA-1(KI) GATA-2(KI/KI) mutant mice display complete megakaryopoietic failure, a phenocopy of FOG-1(-/-) mice. We conclude that FOG-1 requires an interaction with either GATA-1 or -2 as part of its essential role in early megakaryopoiesis. On the basis of these and previous reports, we infer that GATA factor dependence is a critical aspect of FOG protein function. PMID- 12077324 TI - Chimeric snRNA molecules carrying antisense sequences against the splice junctions of exon 51 of the dystrophin pre-mRNA induce exon skipping and restoration of a dystrophin synthesis in Delta 48-50 DMD cells. AB - Deletions and point mutations in the dystrophin gene cause either the severe progressive myopathy Duchenne muscular dystrophy (DMD) or the milder Becker muscular dystrophy, depending on whether the translational reading frame is lost or maintained. Because internal in-frame deletions in the protein produce only mild myopathic symptoms, it should be possible, by preventing the inclusion of specific mutated exon(s) in the mature dystrophin mRNA, to restore a partially corrected phenotype. Such control has been previously accomplished by the use of synthetic oligonucleotides; nevertheless, a significant drawback to this approach is caused by the fact that oligonucleotides would require periodic administrations. To circumvent this problem, we have produced several constructs able to express in vivo, in a stable fashion, large amounts of chimeric RNAs containing antisense sequences. In this paper we show that antisense molecules against exon 51 splice junctions are able to direct skipping of this exon in the human DMD deletion 48-50 and to rescue dystrophin synthesis. We also show that the highest skipping activity was found when antisense constructs against the 5' and 3' splice sites are coexpressed in the same cell. PMID- 12077325 TI - Maternal transmission of P element transposase activity in Drosophila melanogaster depends on the last P intron. AB - Maternal transmission of RNAs or proteins through the egg cytoplasm plays an important role in eukaryotic development. We show that the transposase activity encoded by the P transposable element of Drosophila melanogaster is transmitted through the oocytes of females heterozygous for this element even when these oocytes do not carry the element itself. However, this maternal transmission is abolished when the last of three introns is removed from the P element. These facts imply that maternal transmission of transposase activity involves the RNA transcribed from the P element rather than the polypeptide it encodes, and that to be transmitted maternally, this RNA must possess the last intron. Examination of the intron's sequence reveals that it contains a motif of nine nucleotides that has been implicated in the maternal transmission of developmentally significant RNAs. This same intron limits expression of the P transposase to the germ line of Drosophila. Thus, the last P intron has two important biological functions. PMID- 12077326 TI - The SIN3/RPD3 deacetylase complex is essential for G(2) phase cell cycle progression and regulation of SMRTER corepressor levels. AB - The SIN3 corepressor and RPD3 histone deacetylase are components of the evolutionarily conserved SIN3/RPD3 transcriptional repression complex. Here we show that the SIN3/RPD3 complex and the corepressor SMRTER are required for Drosophila G(2) phase cell cycle progression. Loss of the SIN3, but not the p55, SAP18, or SAP30, component of the SIN3/RPD3 complex by RNA interference (RNAi) causes a cell cycle delay prior to initiation of mitosis. Loss of RPD3 reduces the growth rate of cells but does not cause a distinct cell cycle defect, suggesting that cells are delayed in multiple phases of the cell cycle, including G(2). Thus, the role of the SIN3/RPD3 complex in G(2) phase progression appears to be independent of p55, SAP18, and SAP30. SMRTER protein levels are reduced in SIN3 and RPD3 RNAi cells, and loss of SMRTER by RNAi is sufficient to cause a G(2) phase delay, demonstrating that regulation of SMRTER protein levels by the SIN3/RPD3 complex is a vital component of the transcriptional repression mechanism. Loss of SIN3 does not affect global acetylation of histones H3 and H4, suggesting that the G(2) phase delay is due not to global changes in genome integrity but rather to derepression of SIN3 target genes. PMID- 12077327 TI - Predisposition to arrhythmia and autonomic dysfunction in Nhlh1-deficient mice. AB - Nhlh1 is a basic helix-loop-helix transcription factor whose expression is restricted to the nervous system and which may play a role in neuronal differentiation. To directly study Nhlh1 function, we generated null mice. Homozygous mutant mice were predisposed to premature, adult-onset, unexpected death. Electrocardiograms revealed decreased total heart rate variability, stress induced arrhythmia, and impaired baroreceptor sensitivity. This predisposition to arrhythmia is a likely cause of the observed death in the mutant mice. Heterozygosity for the closely related transcription factor Nhlh2 increased the severity of the Nhlh1-null phenotype. No signs of primary cardiac structural or conduction abnormalities could be detected upon necropsy of the null mice. The pattern of altered heart rhythm observed in basal and experimental conditions (stress and pharmacologically induced) suggests that a deficient parasympathetic tone may contribute to the arrhythmia in the Nhlh1-null mouse. The expression of Nhlh1 in the developing brain stem and in the vagal nuclei in the wild-type mouse further supports this hypothesis. The Nhlh1 mutant mouse may thus provide a model to investigate the contribution of the autonomic nervous system to arrhythmogenesis. PMID- 12077328 TI - 14-3-3 antagonizes Ras-mediated Raf-1 recruitment to the plasma membrane to maintain signaling fidelity. AB - We have investigated the role that S259 phosphorylation, S621 phosphorylation, and 14-3-3 binding play in regulating Raf-1 activity. We show that 14-3-3 binding, rather than Raf-1 phosphorylation, is required for the correct regulation of kinase activity. Phosphorylation of S621 is not required for activity, but 14-3-3 binding is essential. When 14-3-3 binding to conserved region 2 (CR2) was disrupted, Raf-1 basal kinase activity was elevated and it could be further activated by (V12,G37)Ras, (V23)TC21, and (V38)R-Ras. Disruption of 14-3-3 binding at CR2 did not recover binding of Raf-1 to (V12,G37)Ras but allowed more efficient recruitment of Raf-1 to the plasma membrane and stimulated its phosphorylation on S338. Finally, (V12)Ras, but not (V12,G37)Ras, displaced 14-3-3 from full-length Raf-1 and the Raf-1 bound to Ras. GTP was still phosphorylated on S259. Our data suggest that stable association of Raf-1 with the plasma membrane requires Ras-mediated displacement of 14-3-3 from CR2. Small G proteins that cannot displace 14-3-3 fail to recruit Raf-1 to the membrane efficiently and so fail to stimulate kinase activity. PMID- 12077329 TI - CUL-4A is critical for early embryonic development. AB - Ubiquitin-mediated degradation targets cell cycle regulators for proteolysis. Much of the ubiquitin pathway's substrate specificity is conferred by E3 ubiquitin ligases, and cullins are core components of some E3s. CUL-4A encodes one of six mammalian cullins and is amplified and/or overexpressed in breast cancer, which suggests a role in regulating cell cycle progression. To examine CUL-4A's physiologic function, we generated a CUL-4A deletion mutation in mice. No viable CUL-4A(-/-) pups and no homozygous mutant embryos as early as 7.5 days postcoitum (dpc) were recovered. However, CUL-4A(-/-) blastocysts are viable, hatch, form an inner cell mass and trophectoderm, and implant (roughly 4.5 dpc), indicating that CUL-4A(-/-) embryos die between 4.5 and 7.5 dpc. Despite 87% similarity between the Cul-4A and Cul-4B cullins, the CUL-4A(-/-) lethal phenotype indicates that CUL-4A has one or more distinct function(s). Surprisingly, 44% fewer heterozygous pups were recovered than expected by Mendelian genetics, indicating that many heterozygous embryos also die during gestation due to haploinsufficiency. Taken together, our findings indicate that appropriate CUL-4A expression is critical for early embryonic development. PMID- 12077330 TI - The absence of Tssc6, a member of the tetraspanin superfamily, does not affect lymphoid development but enhances in vitro T-cell proliferative responses. AB - The tetraspanins are a family of integral membrane proteins with four transmembrane domains. These molecules form multimolecular networks on the surfaces of many different cell types. Gene-targeting studies have revealed a role for tetraspanins in B- and T-lymphocyte function. We have isolated and deleted a novel tetraspanin, Tssc6, which is expressed exclusively in hematopoietic and lymphoid organs. Using a gene-trapping strategy, we generated an embryonic stem (ES) cell line with an insertion in the Tssc6 locus. Mice were derived from these ES cells and, using RNase protection and reverse transcription PCR, we demonstrated that the insertion resulted in a null mutation of the Tssc6 allele. Mice homozygous for the gene trap insertion (Tssc6(gt/gt) mice) were viable and fertile, with normal steady-state hematopoiesis. Furthermore, responses to hemolysis and granulocyte colony-stimulating factor-induced granulopoiesis were equivalent to those of wild-type mice. Lymphoid development was normal in Tssc6(gt/gt) mice. Whereas Tssc6(gt/gt) B cells responded normally to lipopolysaccharide, anti-CD40, and anti-immunoglobulin M stimulation, Tssc6(gt/gt) T cells showed enhanced responses to concanavalin A, anti-CD3, and anti-CD28. This increased proliferation by Tssc6-deleted T lymphocytes was due to increased interleukin 2 production following T-cell receptor stimulation. These results demonstrate that Tssc6 is not required for normal development of the hematopoietic system but may play a role in the negative regulation of peripheral T-lymphocyte proliferation. PMID- 12077331 TI - The class II transactivator requires brahma-related gene 1 to activate transcription of major histocompatibility complex class II genes. AB - The class II transactivator (CIITA) is the key regulator of major histocompatibility complex (MHC) class II gene transcription. We demonstrate here that CIITA requires the ATPase subunit of an hSWI/SNF complex, brahma-related gene 1 (BRG-1), to activate transcription. When introduced into a cell line lacking BRG-1, CIITA was unable to activate cellular MHC class II genes. Reexpression of the wild-type but not an ATP-binding-deficient BRG-1 protein in this cell line restored the ability of CIITA to transactivate transcription of MHC class II genes. Interestingly, when the activity of CIITA was assayed in the BRG-1-deficient cell line by using a plasmid-based reporter assay, BRG-1 was not required for transcriptional activation, suggesting that the chromatin structure on the plasmid is such that BRG-1 is not necessary. Coimmunoprecipitation experiments were performed to determine if BRG-1 and CIITA proteins associate with each other in cells. We found that the two proteins coimmunoprecipitate and that amino acids 1 to 140 of CIITA are sufficient for binding. Taken together, these data suggest that BRG-1 and, very likely, an hSWI/SNF complex are required for transcription of MHC class II genes. The complex is likely recruited to MHC class II promoters, at least in part, by interaction with CIITA. PMID- 12077332 TI - Neither agouti-related protein nor neuropeptide Y is critically required for the regulation of energy homeostasis in mice. AB - Agouti-related protein (AgRP), a neuropeptide abundantly expressed in the arcuate nucleus of the hypothalamus, potently stimulates feeding and body weight gain in rodents. AgRP is believed to exert its effects through the blockade of signaling by alpha-melanocyte-stimulating hormone at central nervous system (CNS) melanocortin-3 receptor (Mc3r) and Mc4r. We generated AgRP-deficient (Agrp(-/-)) mice to examine the physiological role of AgRP. Agrp(-/-) mice are viable and exhibit normal locomotor activity, growth rates, body composition, and food intake. Additionally, Agrp(-/-) mice display normal responses to starvation, diet induced obesity, and the administration of exogenous leptin or neuropeptide Y (NPY). In situ hybridization failed to detect altered CNS expression levels for proopiomelanocortin, Mc3r, Mc4r, or NPY mRNAs in Agrp(-/-) mice. As AgRP and the orexigenic peptide NPY are coexpressed in neurons of the arcuate nucleus, we generated AgRP and NPY double-knockout (Agrp(-/-);Npy(-/-)) mice to determine whether NPY or AgRP plays a compensatory role in Agrp(-/-) or NPY-deficient (Npy( /-)) mice, respectively. Similarly to mice deficient in either AgRP or NPY, Agrp( /-);Npy(-/-) mice suffer no obvious feeding or body weight deficits and maintain a normal response to starvation. Our results demonstrate that neither AgRP nor NPY is a critically required orexigenic factor, suggesting that other pathways capable of regulating energy homeostasis can compensate for the loss of both AgRP and NPY. PMID- 12077334 TI - Yng1p modulates the activity of Sas3p as a component of the yeast NuA3 Hhistone acetyltransferase complex. AB - The mammalian ING1 gene encodes a tumor suppressor required for the function of p53. In this study we report a novel function for YNG1, a yeast homolog of ING1. Yng1p is a stable component of the NuA3 histone acetyltransferase complex, which contains Sas3p, the yeast homolog of the mammalian MOZ proto-oncogene product, as its catalytic subunit. Yng1p is required for NuA3 function in vivo but surprisingly is not required for the integrity of the complex. Instead, we find that Yng1p mediates the interaction of Sas3p with nucleosomes and is thus required for the ability of NuA3 to modify histone tails. These data, and the observations that other ING1 homologs are found in additional yeast complexes that posttranslationally modify histones, suggest that members of the ING1 class of proteins may have broad roles in enhancing or modifying the activities of chromatin-modifying complexes, thereby regulating their activities in transcription control. PMID- 12077333 TI - The ETS domain transcription factor Elk-1 contains a novel class of repression domain. AB - The ETS domain transcription factor Elk-1 serves as an integration point for different mitogen-activated protein (MAP) kinase pathways. Phosphorylation of Elk 1 by MAP kinases triggers its activation. However, while the activation process is well understood, its downregulation-inactivation is less well characterized. The ETS DNA-binding domain plays a role in the downregulation of Elk-dependent promoter activity following mitogenic activation by recruiting the mSin3A-HDAC complex. Here we have identified a novel evolutionarily conserved repression domain in Elk-1, termed the R motif, which serves to reduce the basal transcriptional activity of Elk-1 and dampen its response to mitogenic signals. This domain is highly potent and portable and can repress transcription in trans. The R motif is related to the CRD1 repression domain in p300 and can functionally replace this domain and confer p21(waf1/cip1) inducibility on p300. However, the R motif acts in a context-dependent manner and is not p21(waf1/cip1) responsive in Elk-1. Thus, the Elk-1 R motif and the p300 CRD1 motif represent a new class of repression domains that are regulated in a context-dependent manner. PMID- 12077335 TI - TRRAP-dependent and TRRAP-independent transcriptional activation by Myc family oncoproteins. AB - We demonstrate that transformation-transactivation domain-associated protein (TRRAP) binding and the recruitment of histone H3 and H4 acetyltransferase activities are required for the transactivation of a silent telomerase reverse transcriptase (TERT) gene in exponentially growing human fibroblasts by c-Myc or N-Myc protein. However, recruitment of TRRAP by c- or N-Myc is dispensable for the partial induction of several basally expressed genes in exponentially growing primary and immortalized fibroblasts. Furthermore, recruitment of TRRAP is required for c-Myc- or N-Myc-mediated oncogenic transformation but not for the partial restoration of the growth defect in myc-null fibroblasts. A segment of the adenovirus E1A protein fused to a transformation-defective N-Myc protein carrying a small deletion in the transactivation domain specifically restores interaction with TRRAP, activates the silent TERT gene, induces acetylation of histones H3 and H4 at the TERT promoter, and transforms primary cells. Accordingly, wild-type L-Myc is much less efficient in TRRAP binding, activation of the silent TERT gene, and transformation of primary fibroblasts. Nevertheless, L-Myc is a potent activator of several basally expressed genes and can fully restore the growth defect of myc-null cells. These results suggest a differential requirement for TRRAP for several Myc-mediated activities. PMID- 12077336 TI - Nir2, a human homolog of Drosophila melanogaster retinal degeneration B protein, is essential for cytokinesis. AB - Cytokinesis, the final stage of eukaryotic cell division, ensures the production of two daughter cells. It requires fine coordination between the plasma membrane and cytoskeletal networks, and it is known to be regulated by several intracellular proteins, including the small GTPase Rho and its effectors. In this study we provide evidence that the protein Nir2 is essential for cytokinesis. Microinjection of anti-Nir2 antibodies into interphase cells blocks cytokinesis, as it results in the production of multinucleate cells. Immunolocalization studies revealed that Nir2 is mainly localized in the Golgi apparatus in interphase cells, but it is recruited to the cleavage furrow and the midbody during cytokinesis. Nir2 colocalizes with the small GTPase RhoA in the cleavage furrow and the midbody, and it associates with RhoA in mitotic cells. Its N terminal region, which contains a phosphatidylinositol transfer domain and a novel Rho-inhibitory domain (Rid), is required for normal cytokinesis, as overexpression of an N-terminal-truncated mutant blocks cytokinesis completion. Time-lapse videomicroscopy revealed that this mutant normally initiates cytokinesis but fails to complete it, due to cleavage furrow regression, while Rid markedly affects cytokinesis due to abnormal contractility. Rid-expressing cells exhibit aberrant ingression and ectopic cleavage sites; the cells fail to segregate into daughter cells and they form a long unseparated bridge-like cytoplasmic structure. These results provide new insight into the cellular functions of Nir2 and introduce it as a novel regulator of cytokinesis. PMID- 12077337 TI - Dissection of a complex phenotype by functional genomics reveals roles for the yeast cyclin-dependent protein kinase Pho85 in stress adaptation and cell integrity. AB - Cyclin-dependent kinases (Cdks) are key regulators of the cell division cycle. Pho85 is a multifunctional Cdk in budding yeast involved in aspects of metabolism, the cell cycle, cell polarity, and gene expression. Consistent with a broad spectrum of functions, Pho85 associates with a family of 10 cyclins and deletion of PHO85 causes a pleiotropic phenotype. Discovering the physiological substrates of protein kinases is a major challenge, and we have pursued a number of genomics approaches to reveal the processes regulated by Pho85 and to understand the root cause of reduced cellular fitness in pho85Delta mutant strains. We used a functional-genomics approach called synthetic genetic array (SGA) analysis to systematically identify strain backgrounds in which PHO85 is required for viability. In parallel, we used DNA microarrays to examine the genome-wide transcriptional consequences of deleting PHO85 or members of the Pho85 cyclin family. Using this pairwise approach coupled with phenotypic tests, we uncovered clear roles for Pho85 in cell integrity and the response to adverse growth conditions. Importantly, our combined approach allowed us to ascribe new aspects of the complex pho85 phenotype to particular cyclins; our data highlight a cell integrity function for the Pcl1,2 subgroup of Pho85 Cdks that is independent of a role for the Pho80-Pho85 kinase in the response to stress. Using a modification of the SGA technique to screen for suppressors of pho85Delta strain growth defects, we found that deletion of putative vacuole protein gene VTC4 suppressed the sensitivity of the pho85Delta strain to elevated CaCl(2) and many other stress conditions. Expression of VTC4 is regulated by Pho4, a transcription factor that is inhibited by the Pho80-Pho85 kinase. Genetic tests and electron microscopy experiments suggest that VTC4 is a key target of Pho4 and that Pho80-Pho85-mediated regulation of VTC4 expression is required for proper vacuole function and for yeast cell survival under a variety of suboptimal conditions. The integration of multiple genomics approaches is likely to be a generally useful strategy for extracting functional information from pleiotropic mutant phenotypes. PMID- 12077338 TI - The Dorsal Rel homology domain plays an active role in transcriptional regulation. AB - The Dorsal morphogen directs formation of the Drosophila dorsoventral axis by both activating and repressing transcription. It contains an N-terminal Rel homology domain (RHD), which is responsible for DNA binding and regulated nuclear import, and a C-terminal domain (CTD) that contains activation and repression motifs. To determine if the RHD has a direct role in transcriptional control, we analyzed a series of RHD mutations in S2 cells and embryos. Two classes of mutations (termed class I and class II mutations) that alter activation without affecting DNA binding or nuclear import were identified. The two classes appear to define distinct protein interaction surfaces on opposite faces of the RHD. Class I mutations enhance an apparently inhibitory interaction between the RHD and the CTD and eliminate both activation and repression by Dorsal. In contrast, class II mutations result in increased activation in S2 cells but severely decreased activation in embryos and have little effect on repression. Analysis of the cuticles of class II mutant embryos suggests that, in the absence of Dorsal mediated activation, Dorsal-mediated repression is not sufficient to pattern the embryo. These results provide some of the first evidence that the RHD plays an active role in transcriptional regulation in intact multicellular organisms. PMID- 12077339 TI - Astrocyte-specific inactivation of the neurofibromatosis 1 gene (NF1) is insufficient for astrocytoma formation. AB - Individuals with the neurofibromatosis 1 (NF1) inherited tumor syndrome develop low-grade gliomas (astrocytomas) at an increased frequency, suggesting that the NF1 gene is a critical growth regulator for astrocytes. In an effort to determine the contribution of the NF1 gene product, neurofibromin, to astrocyte growth regulation and NF1-associated astrocytoma formation, we generated astrocyte specific Nf1 conditional knockout mice (Nf1(GFAP)CKO) by using Cre/LoxP technology. Transgenic mice were developed in which Cre recombinase was specifically expressed in astrocytes by embryonic day 14.5. Successive intercrossing with mice bearing a conditional Nf1 allele (Nf1flox) resulted in GFAP-Cre Nf1flox/flox (Nf1(GFAP)CKO) animals. No astrocytoma formation or neurological impairment was observed in Nf1(GFAP)CKO mice after 20 months, but increased numbers of proliferating astrocytes were observed in several brain regions. To determine the consequence of Nf1 inactivation at different developmental times, the growth properties of embryonic day 12.5 and postnatal day 2 Nf1 null astrocytes were analyzed. Nf1 null astrocytes exhibited increased proliferation but lacked tumorigenic properties in vitro and did not form tumors when injected into immunocompromised mouse brains in vivo. Collectively, our results suggest that loss of neurofibromin is not sufficient for astrocytoma formation in mice and that other genetic or environmental factors might influence NF1-associated glioma tumorigenesis. PMID- 12077340 TI - Selective cooperation between fatty acid binding proteins and peroxisome proliferator-activated receptors in regulating transcription. AB - Lipophilic compounds such as retinoic acid and long-chain fatty acids regulate gene transcription by activating nuclear receptors such as retinoic acid receptors (RARs) and peroxisome proliferator-activated receptors (PPARs). These compounds also bind in cells to members of the family of intracellular lipid binding proteins, which includes cellular retinoic acid-binding proteins (CRABPs) and fatty acid binding proteins (FABPs). We previously reported that CRABP-II enhances the transcriptional activity of RAR by directly targeting retinoic acid to the receptor. Here, potential functional cooperation between FABPs and PPARs in regulating the transcriptional activities of their common ligands was investigated. We show that adipocyte FABP and keratinocyte FABP (A-FABP and K FABP, respectively) selectively enhance the activities of PPARgamma and PPARbeta, respectively, and that these FABPs massively relocate to the nucleus in response to selective ligands for the PPAR isotype which they activate. We show further that A-FABP and K-FABP interact directly with PPARgamma and PPARbeta and that they do so in a receptor- and ligand-selective manner. Finally, the data demonstrate that the presence of high levels of K-FABP in keratinocytes is essential for PPARbeta-mediated induction of differentiation of these cells. Taken together, the data establish that A-FABP and K-FABP govern the transcriptional activities of their ligands by targeting them to cognate PPARs in the nucleus, thereby enabling PPARs to exert their biological functions. PMID- 12077341 TI - H-Ras signaling and K-Ras signaling are differentially dependent on endocytosis. AB - Endocytosis is required for efficient mitogen-activated protein kinase (MAPK) activation by activated growth factor receptors. We examined if H-Ras and K-Ras proteins, which are distributed across different plasma membrane microdomains, have equal access to the endocytic compartment and whether this access is necessary for downstream signaling. Inhibition of endocytosis by dominant interfering dynamin-K44A blocked H-Ras but not K-Ras-mediated PC12 cell differentiation and selectively inhibited H-Ras- but not K-Ras-mediated Raf-1 activation in BHK cells. H-Ras- but not K-Ras-mediated Raf-1 activation was also selectively dependent on phosphoinositide 3-kinase activity. Stimulation of endocytosis and endocytic recycling by wild-type Rab5 potentiated H-Ras-mediated Raf-1 activation. In contrast, Rab5-Q79L, which stimulates endocytosis but not endocytic recycling, redistributed activated H-Ras from the plasma membrane into enlarged endosomes and inhibited H-Ras-mediated Raf-1 activation. Rab5-Q79L expression did not cause the accumulation of wild-type H-Ras in enlarged endosomes. Expression of wild-type Rab5 or Rab5-Q79L increased the specific activity of K-Ras-activated Raf-1 but did not result in any redistribution of K Ras from the plasma membrane to endosomes. These results show that H-Ras but not K-Ras signaling though the Raf/MEK/MAPK cascade requires endocytosis and endocytic recycling. The data also suggest a mechanism for returning Raf-1 to the cytosol after plasma membrane recruitment. PMID- 12077342 TI - Mammalian PRP4 kinase copurifies and interacts with components of both the U5 snRNP and the N-CoR deacetylase complexes. AB - A growing body of evidence supports the coordination of pre-mRNA processing and transcriptional regulation. We demonstrate here that mammalian PRP4 kinase (PRP4K) is associated with complexes involved in both of these processes. PRP4K is implicated in pre-mRNA splicing as the homologue of the Schizosaccharomyces pombe pre-mRNA splicing kinase Prp4p, and it is enriched in SC35-containing nuclear splicing speckles. RNA interference of Caenorhabditis elegans PRP4K indicates that it is essential in metazoans. In support of a role for PRP4K in pre-mRNA splicing, we identified PRP6, SWAP, and pinin as interacting proteins and demonstrated that PRP4K is a U5 snRNP-associated kinase. In addition, BRG1 and N-CoR, components of nuclear hormone coactivator and corepressor complexes, also interact with PRP4K. PRP4K coimmunoprecipitates with N-CoR, BRG1, pinin, and PRP6, and we present data suggesting that PRP6 and BRG1 are substrates of this kinase. Lastly, PRP4K, BRG1, and PRP6 can be purified as components of the N-CoR 2 complex, and affinity-purified PRP4K/N-CoR complexes exhibit deacetylase activity. We suggest that PRP4K is an essential kinase that, in association with the both U5 snRNP and N-CoR deacetylase complexes, demonstrates a possible coordination of pre-mRNA splicing with chromatin remodeling events involved in transcriptional regulation. PMID- 12077344 TI - Generation of mice deficient for macrophage galactose- and N-acetylgalactosamine specific lectin: limited role in lymphoid and erythroid homeostasis and evidence for multiple lectins. AB - Macrophage receptors function in pattern recognition for the induction of innate immunity, in cellular communication to mediate the regulation of adaptive immune responses, and in the clearance of some glycosylated cells or glycoproteins from the circulation. They also function in homeostasis by initiating the engulfment of apoptotic cells. Evidence has suggested that macrophage receptors function to recognize cells that are destined for programmed cell death but not yet overtly apoptotic. We have examined the function of a macrophage receptor specific for unsialylated glycoproteins, known as the mouse macrophage galactose- and N acetylgalactosamine-specific lectin (mMGL) (Ii et al., J. Biol. Chem. 265:11295 11298, 1990; Sato et al., J. Biochem. [Tokyo] 111:331-336, 1992; Yamamoto et al., Biochemistry 33:8159-8166, 1994). With targeted disruption, we tested whether mMGL is necessary for macrophage function, controlled thymic development, the loss of activated CD8 T cells, and the turnover of red blood cells. Evidence indicates that mMGL may play a nonessential role in several of these macrophage functions. Experiments are presented that indicate the existence of another galactose- and N-acetylgalactosamine-recognizing lectin distinct from mMGL. This may explain the absence of a strong phenotype in mMGL-deficient mice. PMID- 12077343 TI - A two-stage, p16(INK4A)- and p53-dependent keratinocyte senescence mechanism that limits replicative potential independent of telomere status. AB - With increasing frequency during serial passage in culture, primary human keratinocytes express p16(INK4A) (p16) and undergo senescence arrest. Keratinocytes engineered to express hTERT maintain long telomeres but typically are not immortalized unless, by mutation or other heritable event, they avoid or greatly reduce p16 expression. We have confirmed that keratinocytes undergo p16 related senescence during growth in culture, whether in the fibroblast feeder cell system or in the specialized K-sfm medium formulation, and that this mechanism can act as a barrier to immortalization following hTERT expression. We have characterized the p16-related arrest mechanism more precisely by interfering specifically with several regulators of cell cycle control. Epidermal, oral mucosal, corneal limbal, and conjunctival keratinocytes were transduced to express a p16-insensitive mutant cdk4 (cdk4(R24C)), to abolish p16 control, and/or a dominant negative mutant p53 (p53DD), to abolish p53 function. Expression of either cdk4(R24C) or p53DD alone had little effect on life span, but expression of both permitted cells to divide 25 to 43 population doublings (PD) beyond their normal limit. Keratinocytes from a p16(+/-) individual transduced to express p53DD alone displayed a 31-PD life span extension associated with selective growth of variants that had lost the wild-type p16 allele. Cells in which both p53 and p16 were nonfunctional divided rapidly during their extended life span but experienced telomere erosion and ultimately ceased growth with very short telomeres. Expression of hTERT in these cells immortalized them. Keratinocytes engineered to express cdk4(R24C) and hTERT but not p53DD did not exhibit an extended life span. Rare immortal variants exhibiting p53 pathway defects arose from them, however, indicating that the p53-dependent component of keratinocyte senescence is telomere independent. Mutational loss of p16 and p53 has been found to be a frequent early event in the development of squamous cell carcinoma. Our results suggest that such mutations endow keratinocytes with extended replicative potential which may serve to increase the probability of neoplastic progression. PMID- 12077345 TI - Drosophila Mi-2 negatively regulates dDREF by inhibiting its DNA-binding activity. AB - Drosophila melanogaster DNA replication-related element (DRE) factor (dDREF) is a transcriptional regulatory factor required for the expression of genes carrying the 5'-TATCGATA DRE. dDREF has been reported to bind to a sequence in the chromatin boundary element, and thus, dDREF may play a part in regulating insulator activity. To generate further insights into dDREF function, we carried out a Saccharomyces cerevisiae two-hybrid screening with DREF polypeptide as bait and identified Mi-2 as a DREF-interacting protein. Biochemical analyses revealed that the C-terminal region of Drosophila Mi-2 (dMi-2) specifically binds to the DNA-binding domain of dDREF. Electrophoretic mobility shift assays showed that dMi-2 thereby inhibits the DNA-binding activity of dDREF. Ectopic expression of dDREF and dMi-2 in eye imaginal discs resulted in severe and mild rough-eye phenotypes, respectively, whereas flies simultaneously expressing both proteins exhibited almost-normal eye phenotypes. Half-dose reduction of the dMi-2 gene enhanced the DREF-induced rough-eye phenotype. Immunostaining of polytene chromosomes of salivary glands showed that dDREF and dMi-2 bind in mutually exclusive ways. These lines of evidence define a novel function of dMi-2 in the negative regulation of dDREF by its DNA-binding activity. Finally, we postulated that dDREF and dMi-2 may demonstrate reciprocal regulation of their functions. PMID- 12077346 TI - Association of DNA polymerase mu (pol mu) with Ku and ligase IV: role for pol mu in end-joining double-strand break repair. AB - Mammalian DNA polymerase mu (pol mu) is related to terminal deoxynucleotidyl transferase, but its biological role is not yet clear. We show here that after exposure of cells to ionizing radiation (IR), levels of pol mu protein increase. pol mu also forms discrete nuclear foci after IR, and these foci are largely coincident with IR-induced foci of gammaH2AX, a previously characterized marker of sites of DNA double-strand breaks. pol mu is thus part of the cellular response to DNA double-strand breaks. pol mu also associates in cell extracts with the nonhomologous end-joining repair factor Ku and requires both Ku and another end-joining factor, XRCC4-ligase IV, to form a stable complex on DNA in vitro. pol mu in turn facilitates both stable recruitment of XRCC4-ligase IV to Ku-bound DNA and ligase IV-dependent end joining. In contrast, the related mammalian DNA polymerase beta does not form a complex with Ku and XRCC4-ligase IV and is less effective than pol mu in facilitating joining mediated by these factors. Our data thus support an important role for pol mu in the end-joining pathway for repair of double-strand breaks. PMID- 12077347 TI - Novel transcription coactivator complex containing activating signal cointegrator 1. AB - Human activating signal cointegrator 1 (hASC-1) was originally isolated as a transcriptional coactivator of nuclear receptors. Here we report that ASC-1 exists as a steady-state complex associated with three polypeptides, P200, P100, and P50, in HeLa nuclei; stimulates transactivation by serum response factor (SRF), activating protein 1 (AP-1), and nuclear factor kappaB (NF-kappaB) through direct binding to SRF, c-Jun, p50, and p65; and relieves the previously described transrepression between nuclear receptors and either AP-1 or NF-kappaB. Interestingly, ectopic expression of Caenorhabditis elegans ASC-1 (ceASC-1), an ASC-1 homologue that binds P200 and P100, like hASC-1, while weakly interacting only with p65, in HeLa cells appears to replace endogenous hASC-1 from the hASC-1 complex and exerts potent dominant-negative effects on AP-1, NF-kappaB, and SRF transactivation. In addition, neutralization of endogenous P50 by single-cell microinjection of a P50 antibody inhibits AP-1 transactivation; the inhibition is relieved by coexpression of wild-type P50, but not of P50DeltaKH, a mutant form that does not interact with P200. Overall, these results suggest that the endogenous hASC-1 complex appears to play an essential role in AP-1, SRF, and NF kappaB transactivation and to mediate the transrepression between nuclear receptors and either AP-1 or NF-kappaB in vivo. PMID- 12077348 TI - Targeted deletion of both thymidine phosphorylase and uridine phosphorylase and consequent disorders in mice. AB - Thymidine phosphorylase (TP) regulates intracellular and plasma thymidine levels. TP deficiency is hypothesized to (i) increase levels of thymidine in plasma, (ii) lead to mitochondrial DNA alterations, and (iii) cause mitochondrial neurogastrointestinal encephalomyopathy (MNGIE). In order to elucidate the physiological roles of TP, we generated mice deficient in the TP gene. Although TP activity in the liver was inhibited in these mice, it was fully maintained in the small intestine. Murine uridine phosphorylase (UP), unlike human UP, cleaves thymidine, as well as uridine. We therefore generated TP-UP double-knockout (TP( /-) UP(-/-)) mice. TP activities were inhibited in TP(-/-) UP(-/-) mice, and the level of thymidine in the plasma of TP(-/-) UP(-/-) mice was higher than for TP( /-) mice. Unexpectedly, we could not observe alterations of mitochondrial DNA or pathological changes in the muscles of the TP(-/-) UP(-/-) mice, even when these mice were fed thymidine for 7 months. However, we did find hyperintense lesions on magnetic resonance T(2) maps in the brain and axonal edema by electron microscopic study of the brain in TP(-/-) UP(-/-) mice. These findings suggested that the inhibition of TP activity caused the elevation of pyrimidine levels in plasma and consequent axonal swelling in the brains of mice. Since lesions in the brain do not appear to be due to mitochondrial alterations and pathological changes in the muscle were not found, this model will provide further insights into the causes of MNGIE. PMID- 12077349 TI - PIAS proteins modulate transcription factors by functioning as SUMO-1 ligases. AB - PIAS (protein inhibitor of activated STAT) proteins interact with and modulate the activities of various transcription factors. In this work, we demonstrate that PIAS proteins xalpha, xbeta, 1, and 3 interact with the small ubiquitin related modifier SUMO-1 and its E2 conjugase, Ubc9, and that PIAS proteins themselves are covalently modified by SUMO-1 (sumoylated). PIAS proteins also tether other sumoylated proteins in a noncovalent fashion. Furthermore, recombinant PIASxalpha enhances Ubc9-mediated sumoylation of the androgen receptor and c-Jun in vitro. Importantly, PIAS proteins differ in their abilities to promote sumoylation in intact cells. The ability to stimulate protein sumoylation and the interaction with sumoylated proteins are dependent on the conserved PIAS RING finger-like domain. These functions are linked to the activity of PIASxalpha on androgen receptor-dependent transcription. Collectively, our results imply that PIAS proteins function as SUMO-1-tethering proteins and zinc finger-dependent E3 SUMO protein ligases, and these properties are likely to explain their ability to modulate the activities of various transcription factors. PMID- 12077352 TI - An invitation for a seminar-- a conspiracy theory. By Caveman. PMID- 12077350 TI - Characterization of mice deficient in the Src family nonreceptor tyrosine kinase Frk/rak. AB - Frk/rak belongs to a novel family of Src kinases with epithelial tissue-specific expression. Although developmental expression patterns and functional overexpression in vitro have associated these kinases with growth suppression and differentiation, their physiological functions remain largely unknown. We therefore generated mice carrying a null mutation in iyk, the mouse homolog of Frk/rak. We report here that frk/rak(-/-) mice are viable, show similar growth rates to wild-type animals, and are fertile. Furthermore, a 2-year study of health and survival did not identify differences in the incidence and spectrum of spontaneous tumors or provide evidence of hyperplasias in frk/rak(-/-) epithelial tissues. Histological analysis of organs failed to reveal any morphological changes in epithelial tissues that normally express high levels of Frk/rak. Ultrastructural analysis of intestinal enterocytes did not identify defects in brush border morphology or structural polarization, demonstrating that Frk/rak is dispensable for intestinal cytodifferentiation. Additionally, frk/rak-null mice do not display altered sensitivity to intestinal damage induced by ionizing radiation. cDNA microarray analysis revealed an increase in c-src expression and identified subtle changes in the expression of genes regulated by thyroid hormones. Significant decreases in the circulating levels of T3 but not T4 hormone are consistent with this observation and reminiscent of euthyroid sick syndrome, a stress-associated clinical condition. PMID- 12077351 TI - Lsm proteins are required for normal processing of pre-tRNAs and their efficient association with La-homologous protein Lhp1p. AB - Depletion of any of the five essential proteins Lsm2p to Lsm5p and Lsm8p leads to strong accumulation of all tested unspliced pre-tRNA species, as well as accumulation of 5' and 3' unprocessed species. Aberrant 3'-extended pre-tRNAs were detected, presumably due to stabilization of transcripts that fail to undergo correct transcription termination, and the accumulation of truncated tRNA fragments was also observed. Tandem affinity purification-tagged Lsm3p was associated with pre-tRNA primary transcripts and, less efficiently, with other unspliced pre-tRNA intermediates but not mature tRNAs. Association of the Saccharomyces cerevisiae La homologue Lhp1p with pre-tRNAs was reduced approximately threefold on depletion of Lsm3p or Lsm5p. The association of Lhp1p with larger RNA polymerase III transcripts, pre-RNase P RNA and the signal recognition particle RNA (scR1), was more drastically reduced. The impaired pre tRNA processing seen on Lsm depletion is not, however, due solely to reduced Lhp1p association as evidenced by analysis of lhp1-Delta strains depleted of Lsm3p or Lsm5p. These data are consistent with roles for an Lsm complex as a chaperone that facilitates the efficient association of pre-tRNA processing factors with their substrates. PMID- 12077354 TI - Vesicle tethering complexes in membrane traffic. AB - Despite the recent progress in the field of membrane traffic, the question of how the specificity of membrane fusion is achieved has yet to be resolved. It has become apparent that the SNARE proteins, although central to the process of fusion, are often not the first point of contact between a vesicle and its target. Instead, a poorly understood tethering process physically links the two before fusion occurs. Many factors that have an apparent role in tethering have been identified. Among these are several large protein complexes. Until recently, these seemed unrelated, which was a surprise since proteins involved in membrane traffic often form families, members of which function in each transport step. Recent work has shown that three of the complexes are in fact related. We refer to these as the 'quatrefoil' tethering complexes, since they appear to share a fourfold nature. Here we describe the quatrefoil complexes and other, unrelated, tethering complexes, and discuss ideas about their function. We propose that vesicle tethering may have separate kinetic and thermodynamic elements and that it may be usefully divided into events upstream and downstream of the function of Rab GTPases. Moreover, the diversity of tethering complexes in the cell suggests that not all tethering events occur through the same mechanisms. PMID- 12077353 TI - Molecular evolution of the actin family. AB - Members of the actin family have well-characterized cytoskeletal functions, but actin and actin-related proteins (ARPs) have also been implicated in nuclear activities. Previous analyses of the actin family have identified four conserved subfamilies, but many actin-related proteins (ARPs) do not fall into these groups. A new systematic phylogenetic analysis reveals that at least eight ARP subfamilies are conserved from humans to yeast, indicating that these ARPs are part of the core set of eukaryotic proteins. Members of at least three subfamilies appear to be involved in chromatin remodeling, suggesting that ARPs play ancient, fundamental roles in this nuclear process. PMID- 12077355 TI - Hyperproliferation, induction of c-Myc and 14-3-3sigma, but no cell fragility in keratin-10-null mice. AB - In the past, keratins have been established as structural proteins. Indeed, mutations in keratin 10 (K10) and other epidermal keratins lead to severe skin fragility syndromes. Here, we present adult K10-/- mice, which reveal a novel connection between the regulation of cell proliferation and K10. Unlike most keratin mutant mice, the epidermis of adult K10-/- mice showed no cytolysis but displayed hyperproliferation of basal keratinocytes and an increased cell size. BrdU labelling revealed a shortened transition time for keratinocytes migrating outwards and DAPI staining of epidermal sheets uncovered an impaired organization of epidermal proliferation units. These remarkable changes were accompanied by the induction of c-Myc, cyclin D1, 14-3-3sigma and of wound healing keratins K6 and K16. The phosphorylation of Rb remained unaltered. In line with the downregulation of K10 in squamous cell carcinomas and its absence in proliferating cells in vivo, our data suggest that the tissue-restricted expression of some members of the keratin gene family not only serves structural functions. Our results imply that the altered composition of the suprabasal cytoskeleton is able to alter the proliferation state of basal cells through the induction of c-Myc. A previous model based on transfection of K10 in immortalized human keratinocytes suggested a direct involvement of K10 in cell cycle control. While those experiments were performed in human cultured keratinocytes, our data establish, that in vivo, K10 acts by an indirect control mechanism in trans. PMID- 12077356 TI - Compartmentation of enzymes in a microbody, the glycosome, is essential in Trypanosoma brucei. AB - All kinetoplastids contain membrane-bound microbodies known as glycosomes, in which several metabolic pathways including part of glycolysis are compartmentalized. Peroxin 2 is essential for the import of many proteins into the microbodies of yeasts and mammals. The PEX2 gene of Trypanosoma brucei was identified and its expression was silenced by means of tetracycline-inducible RNA interference. Bloodstream-form trypanosomes, which rely exclusively on glycolysis for ATP generation, died rapidly upon PEX2 depletion. Insect-form (procyclic) trypanosomes do not rely solely on glycolysis for ATP synthesis. PEX2 depletion in procyclic forms resulted in relocation of most tested matrix proteins to the cytosol, and these mutants also died. Compartmentation of microbody enzymes is therefore essential for survival of bloodstream and procyclic T. brucei. In contrast, yeasts and cultured mammalian cells grow normally in the absence of peroxisomal membranes unless placed on selective media. PMID- 12077357 TI - Midkine, a heparin-binding growth factor, promotes growth and glycosaminoglycan synthesis of endothelial cells through its action on smooth muscle cells in an artificial blood vessel model. AB - To study the interactions between smooth muscle cells and endothelial cells in vitro, we developed an artificial blood vessel model, which consisted of collagen gel containing human aortic smooth muscle cells and human umbilical vein endothelial cells grown on the gel. The blood vessel model was utilized to investigate the role of midkine, a heparin-binding growth factor, in the intercellular interactions that are important in angiogenesis. In the blood vessel model, midkine induced stratification of the endothelial cells and increased their proliferation and glycosaminoglycan synthesis. However, midkine had no effect on the smooth muscle cells or endothelial cells when they were cultured separately. Increased proliferation of the endothelial cells was also attained by coculturing them with smooth muscle cells in the presence of midkine or culturing endothelial cells with the conditioned medium of the smooth muscle cells, which had been treated with midkine. These experiments indicate that the target of midkine was smooth muscle cells, which secreted factor(s) acting on the endothelial cells. We identified interleukin-8 as one such factor; the synthesis of interleukin-8 by the smooth muscle cells was increased by exposure to midkine, and anti-interleukin-8 inhibited the midkine action. Furthermore, interleukin-8 caused stratification of the endothelial cells in the blood vessel model. These results provided evidence that midkine is one of the factors involved in epithelial-mesenchymal interactions. PMID- 12077358 TI - Role of the beta1-integrin cytoplasmic tail in mediating invasin-promoted internalization of Yersinia. AB - Invasin of Yersinia pseudotuberculosis binds to beta1-integrins on host cells and triggers internalization of the bacterium. To elucidate the mechanism behind the beta1-integrin-mediated internalization of Yersinia, a beta1-integrin-deficient cell line, GD25, transfected with wild-type beta1A, beta1B or different mutants of the beta1A subunit was used. Both beta1A and beta1B bound to invasin expressing bacteria, but only beta1A was able to mediate internalization of the bacteria. The cytoplasmic region of beta1A, differing from beta1B, contains two NPXY motifs surrounding a double threonine site. Exchanging the tyrosines of the two NPXYs to phenylalanines did not inhibit the uptake, whereas a marked reduction was seen when the first tyrosine (Y783) was exchanged to alanine. A similar reduction was seen when the two nearby threonines (TT788-9) were exchanged with alanines. It was also noted that cells affected in bacterial internalization exhibited reduced spreading capability when seeded onto invasin, suggesting a correlation between the internalization of invasin-expressing bacteria and invasin-induced spreading. Likewise, integrins defective in forming peripheral focal complex structures was unable to mediate uptake of invasin expressing bacteria. PMID- 12077359 TI - Isolation of human progenitor liver epithelial cells with extensive replication capacity and differentiation into mature hepatocytes. AB - The liver can regenerate itself through the progenitor cells it harbors. Here we demonstrate isolation of epithelial progenitor/stem cells from the fetal human liver, which contains a large number of hepatoblasts. Progenitor liver cells displayed clonogenic capacity, expressed genes observed in hepatocytes, bile duct cells and oval cells, and incorporated genes transferred by adenoviral or lentiviral vectors. Under culture conditions, progenitor cells proliferated for several months, with each cell undergoing more than forty divisions, but they retained normal karyotypes. Progenitor cells differentiated into mature hepatocytes in mice with severe combined immunodeficiency, both when in an ectopic location and when in the liver itself. Cells integrated in the liver parenchyma and proliferated following liver injury. An abundance of progenitor cells in the fetal human liver is consistent with models indicating depletion of progenitor/stem cells during aging and maturation of organs. The studies indicate that isolation of progenitor cells from fetal organs will be appropriate for establishing novel systems to investigate basic mechanisms and for cell and gene therapy. PMID- 12077360 TI - Cas, Fak and Pyk2 function in diverse signaling cascades to promote Yersinia uptake. AB - The interplay between pathogen-encoded virulence factors and host cell signaling networks is critical for both the establishment and clearance of microbial infections. Yersinia uptake into host cells serves as an in vitro model for exploring how host cells respond to Yersinia adherence. In this study, we provide insight into the molecular nature and regulation of signaling networks that contribute to the uptake process. Using a reconstitution approach in Fak(-/-) fibroblasts, we have been able to specifically address the interplay between Fak, Cas and Pyk2 in this process. We show that both Fak and Cas play roles in the Yersinia uptake process and that Cas can function in a novel pathway that is independent of Fak. Fak-dependent Yersinia uptake does not appear to involve Cas Crk signaling. By contrast, Cas-mediated uptake in the absence of Fak requires Crk as well as the protein tyrosine kinases Pyk2 and Src. In spite of these differences, the requirement for Rac1 activity is a common feature of both pathways. Furthermore, blocking the function of either Fak or Cas induces similar morphological defects in Yersinia internalization, which are manifested by incomplete membrane protrusive activity that is consistent with an inhibition of Rac1 activity. Pyk2 also functions in Yersinia uptake by macrophages, which are physiologically important for clearing Yersinia infections. Taken together, these data provide new insight into the host cellular signaling networks that are initiated upon infection with Y. pseudotuberculosis. Importantly, these findings also contribute to a better understanding of other cellular processes that involve actin remodeling, including the host response to other microbial pathogens, cell adhesion and migration. PMID- 12077361 TI - Tristetraprolin and LPS-inducible CXC chemokine are rapidly induced in presumptive satellite cells in response to skeletal muscle injury. AB - Myogenic precursor cells known as satellite cells persist in adult skeletal muscle and are responsible for its ability to regenerate after injury. Quiescent satellite cells are activated by signals emanating from damaged muscle. Here we describe the rapid activation of two genes in response to muscle injury; these transcripts encode LPS-inducible CXC chemokine (LIX), a neutrophil chemoattractant, and Tristetraprolin (TTP), an RNA-binding protein implicated in the regulation of cytokine expression. Using a synchronized cell culture model we show that C2C12 myoblasts arrested in G0 exhibit some molecular attributes of satellite cells in vivo: suppression of MyoD and Myf5 expression during G0 and their reactivation in G1. Synchronization also revealed cell cycle dependent expression of CD34, M-cadherin, HGF and PEA3, genes implicated in satellite cell biology. To identify other genes induced in synchronized C2C12 myoblasts we used differential display PCR and isolated LIX and TTP cDNAs. Both LIX and TTP mRNAs are short-lived, encode molecules implicated in inflammation and are transiently induced during growth activation in vitro. Further, LIX and TTP are rapidly induced in response to muscle damage in vivo. TTP expression precedes that of MyoD and is detected 30 minutes after injury. The spatial distribution of LIX and TTP transcripts in injured muscle suggests expression by satellite cells. Our studies suggest that in addition to generating new cells for repair, activated satellite cells may be a source of signaling molecules involved in tissue remodeling during regeneration. PMID- 12077362 TI - Downregulated AP-1 activity is associated with inhibition of Protein-Kinase-C dependent CD44 and ezrin localisation and upregulation of PKC theta in A431 cells. AB - Progression to an invasive, metastatic tumour requires the coordinated expression and function of a number of gene products, as well as their regulation in the context of invasion. The transcription factor AP-1 regulates expression of many of those genes necessary for implementation of the invasion programme. Two such gene products, CD44 and ezrin, are both upregulated in fibroblasts transformed by v-fos and are commonly implicated in cell motility and invasion. Here we report that CD44 and ezrin colocalise to membrane ruffles and microvilli of A431 cells after treatment with EGF. However, A431 cells expressing dominant-negative c-Jun (TAM67), and which as a consequence fail to invade in response to EGF, also fail to correctly localise CD44 and ezrin. CD44 and ezrin are both substrates for Protein Kinase C, and we show that their EGF-dependent colocalisation requires Protein Kinase C activity. Associated with TAM67 expression and disrupted CD44 and ezrin colocalisation is the increased expression and activation of the novel PKC theta isoform. Expression of PKC theta in A431 cells results in the inhibition of cell motility and disrupted localisation of CD44 and ezrin. We propose that AP-1 regulates the integrity of Protein Kinase C signalling and identifies PKC theta as a potential suppressor of the invasion programme. PMID- 12077363 TI - NBD-labeled phosphatidylcholine enters the yeast vacuole via the pre-vacuolar compartment. AB - At low temperature, the short-chain fluorescent-labeled phospholipids, 1 myristoyl-2-[6-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) aminocaproyl] phosphatidylcholine (M-C6-NBD-PC) and its phosphatidylethanolamine analog, M-C6 NBD-PE, are internalized by flip across the plasma membrane of S. cerevisiae and show similar enrichment in intracellular membranes including the mitochondria and nuclear envelope/ER. At higher temperatures (24-37 degrees C), or if low temperature internalization is followed by warming, M-C6-NBD-PC, but not M-C6-NBD PE, is trafficked to the lumen of the vacuole. Sorting of M-C6-NBD-PC to the vacuole is blocked by energy-depletion and by null mutations in the VPS4 and VPS28 genes required for vesicular traffic from the pre-vacuolar compartment (PVC) to the vacuole. This sorting is not blocked by a temperature-sensitive mutation in SEC12, which inhibits ER to Golgi transport, a null mutation in VPS8, which inhibits Golgi to PVC transport, or temperature-sensitive and null mutations in END4, which inhibit endocytosis from the plasma membrane. Monomethylation or dimethylation of the primary amine head-group of M-C6-NBD-PE is sufficient for sorting to the yeast vacuole in both wild-type yeast and in strains defective in the phosphatidylethanolamine methylation pathway. These data indicate that methylation of M-C6-NBD-PE produces the crucial structural component required to sort these phospholipid analogues to the vacuole via the PVC. PMID- 12077364 TI - Expression of BRG1, a human SWI/SNF component, affects the organisation of actin filaments through the RhoA signalling pathway. AB - The human BRG1 (brahma-related gene 1) protein is a component of the SWI/SNF family of the ATP-dependent chromatin remodelling complexes. We show here that expression of the BRG1 protein, but not of an ATPase-deficient BRG1 protein, in BRG1-deficient SW13 cells alters the organisation of actin filaments. BRG1 expression induces the formation of thick actin filament bundles resembling stress-fibres, structures that are rarely seen in native SW13 cells. BRG1 expression does not influence the activity state of the RhoA-GTPase, which is involved in stress-fibre formation. We find that RhoA is equally activated by stimuli, such as serum, in BRG1-expressing cells, ATPase-deficient BRG1 expressing cells and native SW13 cells. However, the activation of RhoA by lysophosphatidic acid and serum does not trigger the formation of stress-fibre like structures in SW13 cells. Activation of the RhoA-GTPase in BRG1-expressing cells induces stress-fibre-like structures, indicating that the BRG1 can couple RhoA activation to stress-fibre formation. At least two downstream effectors are involved in stress-fibre formation, Rho-kinase/ROCK and Dia. BRG1 expression, but not the expression of the ATP-deficient BRG1, increases the protein level of ROCK1, one form of the Rho-kinase/ROCK. That this is of importance is supported by the findings that an increased Rho-kinase/ROCK activity in SW13 cells, obtained by overexpressing wild-type ROCK1 and ROCK2, induces stress-fibre formation. No specificity between the two Rho-kinase/ROCK forms exists. Our results suggest that the BRG1 protein affects the RhoA pathway by increasing the protein level of ROCK1, which allows stress-fibre-like structures to form. PMID- 12077365 TI - Alterations in the lens capsule contribute to cataractogenesis in SPARC-null mice. AB - The lens capsule, which is also called the lens basement membrane, is a specialized extracellular matrix produced anteriorly by the lens epithelium and posteriorly by newly differentiated fiber cells. SPARC (secreted protein, acidic and rich in cysteine) is a matricellular glycoprotein that regulates cell-cell and cell-matrix interactions, cellular proliferation and differentiation, and the expression of genes encoding extracellular matrix components. SPARC-null mice exhibit lens opacity 1 month after birth and mature cataract and capsular rupture at 5-7 months. In this study, we report disruption of the structural integrity of the lens capsule in mice lacking SPARC. The major structural protein of basement membrane, collagen type IV, in the lens capsule was substantially altered in the absence of SPARC. The lens cells immediately beneath the capsule showed aberrant morphology, with numerous protrusions into the lens basement membrane. SPARC-null lenses at 1 month of age exhibited an increased penetration of dye or radioactive tracer through the capsule, as well as a higher content of water than their wild type counterparts. Moreover, SPARC-null fibers exhibited swelling as early as 1 month of age; by 3 months, all the fiber cells appeared swollen to a marked degree. By contrast, the absence of SPARC had no apparent morphological effect on the early stages of lens formation, cell proliferation or fiber cell differentiation. Degradation of crystallins and MIP 26, or changes in the levels of these proteins, were not detected. These results underscore the importance of the capsular extracellular matrix in the maintenance of lens transparency and indicate that SPARC participates in the synthesis, assembly and/or stabilization of the lens basement membrane. PMID- 12077366 TI - Apoptotic crosstalk of TNF receptors: TNF-R2-induces depletion of TRAF2 and IAP proteins and accelerates TNF-R1-dependent activation of caspase-8. AB - We have recently shown that stimulation of TNF-R2 selectively enhances apoptosis induction by the death receptor TNF-R1. Here, we demonstrate that stimulation of CD30 or CD40 also leads to selective enhancement of TNF-R1-induced cell death. Enhancement of apoptosis was correlated with the depletion of endogenous TRAF2 within 1 to 6 hours. Selective prestimulation of TNF-R2 for several hours inhibited TNF-R2-induced activation of the anti-apoptotic NF-kappaB pathway up to 90% and dramatically enhanced apoptosis induction by this receptor. When both TNF receptors were stimulated simultaneously, TNF-R1-induced NF-kappaB activation remained unaffected but TNF-R1-induced apoptosis was still significantly enhanced. Compared with FasL-induced cell death TNF-R1-induced activation of caspase-8 was significantly weaker and delayed. Costimulation or prestimulation of TNF-R2 enhanced caspase-8 processing. Life cell imaging and confocal microscopy revealed that both TNF-R1 and TNF-R2 recruited the anti-apoptotic factor cIAP1 in a TRAF2-dependent manner. Thus, TNF-R2 may compete with TNF-R1 for the recruitment of newly synthesized TRAF2-bound anti-apoptotic factors, thereby promoting the formation of a caspase-8-activating TNF-R1 complex. Hence, TNF-R2 triggering can interfere with TNF-R1-induced apoptosis by inhibition of NF kappaB-dependent production of anti-apoptotic factors and by blocking the action of anti-apoptotic factors at the post-transcriptional level. PMID- 12077367 TI - Regulation of S33/S37 phosphorylated beta-catenin in normal and transformed cells. AB - A novel phosphorylation-specific antibody (alphapbeta-catenin) was generated against a peptide corresponding to amino acids 33-45 of human beta-catenin, which contained phosphorylated serines at positions 33 and 37. This antibody is specific to phosphorylated beta-catenin and reacts neither with the non phosphorylated protein nor with phosphorylated or non-phosphorylated plakoglobin. It weakly interacts with S33Y beta-catenin but not with the S37A mutant. pbeta catenin is hardly detectable in normal cultured cells and accumulates (up to 55% of total beta-catenin) upon overexpression of the protein or after blocking its degradation by the proteasome. Inhibition of both GSK-3beta and the proteasome resulted in a rapid (t1/2=10 minutes) and reversible reduction in pbeta-catenin levels, suggesting that the protein can undergo dephosphorylation in live cells, at a rate comparable to its phosphorylation by GSK-3beta. pbeta-catenin interacts with LEF-1, but fails to form a ternary complex with DNA, suggesting that it is transcriptionally inactive. Immunofluorescence microscopy indicated that pbeta catenin accumulates in the nuclei of MDCK and BCAP cells when overexpressed and is transiently associated with adherens junctions shortly after their formation. pbeta-catenin only weakly interacts with co-transfected N-cadherin, although it forms a complex with the ubiquitin ligase component beta-TrCP. SW480 colon cancer cells that express a truncated APC, at position 1338, contain high levels of pbeta-catenin, whereas HT29 cells, expressing APC truncated at position 1555, accumulate non-phosphorylated beta-catenin, suggesting that the 1338-1555 amino acid region of APC is involved in the differential regulation of the dephosphorylation and degradation of pbeta-catenin. PMID- 12077368 TI - Cell adhesion differentially regulates the nucleocytoplasmic distribution of active MAP kinases. AB - Cells decide whether to undergo processes, such as proliferation, differentiation and apoptosis, based upon the cues they receive from both circulating factors and integrin-mediated adhesion to the extracellular matrix. Integrins control the activation of the early signaling pathways. For example, growth factor activation of the ERK cascade is enhanced when cells are adherent. In addition, adhesion receptors oversee the cellular localization of critical signaling components. We have recently shown that ERK signaling to the nucleus is regulated by cell adhesion at the level of nucleocytoplasmic trafficking. Since the ERKs are only one class of MAP kinase, we extended these studies to include both JNK and p38 MAP kinases. We have rendered JNK and p38 activation in NIH 3T3 fibroblasts anchorage-independent either by treatment with anisomycin or by expression of upstream activators. Under conditions whereby JNK activation is anchorage independent, we show that localization of JNK to the nucleus and JNK-mediated phosphorylation of c-Jun and Elk-1 is not altered by loss of adhesion. Likewise, the ability of activated p38 to accumulate in the nucleus was similar in suspended and adherent cells. Finally, we show that expression of a form of ERK, which is activated and resistant to nuclear export, reverses the adhesion dependency of ERK phosphorylation of Elk-1. Thus, adhesion differentially regulates the nucleocytoplasmic distribution of MAP kinase members; ERK accumulation in the nucleus occurs more efficiently in adherent cells, whereas nuclear accumulation of active p38 and active JNK are unaffected by changes in adhesion. PMID- 12077370 TI - Changes in tropical clouds and radiation. PMID- 12077371 TI - Measuring success in assisted reproductive technology. PMID- 12077369 TI - Ca2+-induced changes in SNAREs and synaptotagmin I correlate with triggered exocytosis from chromaffin cells: insights gleaned into the signal transduction using trypsin and botulinum toxins. AB - Ca2+-triggered catecholamine exocytosis from chromaffin cells involves SNAP-25, synaptobrevin and syntaxin (known as SNAREs). Synaptotagmin I has been implicated as the Ca2+-sensor because it binds Ca2+, and this enhances its binding to syntaxin, SNAP-25 and phospholipids in vitro. However, most of these interactions are only mediated by [Ca2+]i two orders of magnitude higher than that needed to elicit secretion. Thus, the Ca2+ sensitivities of synaptotagmin I and the other SNAREs were quantified in situ. Secretion elicited from permeabilised cells by microM Ca2+ was accompanied, with almost identical Ca2+ dependencies, by changes in synaptotagmin I, SNAP-25, syntaxin and synaptobrevin that rendered them less susceptible to trypsin. The majority of the trypsin-resistant SNAREs were not associated with SDS-resistant complexes. None of these proteins acquired trypsin resistance in cells rendered incompetent for exocytosis by run-down. Removal of nine C-terminal residues from SNAP-25 by botulinum toxin A reduced both exocytosis and the SNAREs' acquisition of trypsin resistance but did not alter the Ca2+ sensitivity, except for synaptotagmin I. Even after synaptobrevin had been cleaved by botulinum toxin B, all the other proteins still responded to Ca2+. These data support a model whereby Ca2+ is sensed, probably by synaptotagmin I, and the signal passed to syntaxin and SNAP-25 before they interact with synaptobrevin. PMID- 12077372 TI - Biowarfare. Did bioweapons test cause a deadly smallpox outbreak? PMID- 12077373 TI - Cloning. Moratorium replaces ban as U.S. target. PMID- 12077374 TI - Radiological terrorism. New effort aims to thwart dirty bombers. PMID- 12077375 TI - Germany. Gruss takes Max Planck helm. PMID- 12077376 TI - Immunology. Plant a few cells, sprout a thymus. PMID- 12077377 TI - Developmental biology. Nerves tell arteries to make like a tree. PMID- 12077378 TI - Canada. Act seen as first step in protecting species. PMID- 12077379 TI - Canada. Amgen splits with lab, but its money lingers. PMID- 12077380 TI - Exoplanets. Jupiter's brother joins the family. PMID- 12077381 TI - Japan. New program to aid smaller universities. PMID- 12077383 TI - Stem cells. Plasticity: time for a reappraisal? PMID- 12077382 TI - Gene transcription. Demolition crew gets a hand from chaperones. PMID- 12077384 TI - Climate change. Russia can save Kyoto, if it can do the math. PMID- 12077385 TI - Neuroscience. A generation gap in brain activity. PMID- 12077386 TI - Astrophysics. Physicists prepare to catch cosmic bullets. PMID- 12077387 TI - Public health. Worldwide occurrences of arsenic in ground water. PMID- 12077388 TI - Public health. Arsenic epidemiology and drinking water standards. PMID- 12077389 TI - Neurodegeneration. A glutamine-rich trail leads to transcription factors. PMID- 12077390 TI - Planetary science. Where have all the comets gone? PMID- 12077391 TI - Brown dwarfs. Failed stars or overachieving planets? PMID- 12077392 TI - Atom optics. Continuous progress on atom lasers. PMID- 12077394 TI - Climate warming and disease risks for terrestrial and marine biota. AB - Infectious diseases can cause rapid population declines or species extinctions. Many pathogens of terrestrial and marine taxa are sensitive to temperature, rainfall, and humidity, creating synergisms that could affect biodiversity. Climate warming can increase pathogen development and survival rates, disease transmission, and host susceptibility. Although most host-parasite systems are predicted to experience more frequent or severe disease impacts with warming, a subset of pathogens might decline with warming, releasing hosts from disease. Recently, changes in El Nino-Southern Oscillation events have had a detectable influence on marine and terrestrial pathogens, including coral diseases, oyster pathogens, crop pathogens, Rift Valley fever, and human cholera. To improve our ability to predict epidemics in wild populations, it will be necessary to separate the independent and interactive effects of multiple climate drivers on disease impact. PMID- 12077393 TI - Biochemistry. Intramembrane proteases--mixing oil and water. PMID- 12077396 TI - Sorting out chromosome errors. PMID- 12077397 TI - Quirks of fetal environment felt decades later. PMID- 12077398 TI - Cells exchanged during pregnancy live on. PMID- 12077399 TI - Research on contraception still in the doldrums. PMID- 12077400 TI - Germline stem cell transplantation and transgenesis. AB - The recently developed testis cell transplantation method provides a powerful approach to studying the biology of the male germline stem cell and its microenvironment, the stem cell niche. The technique also is being used to examine spermatogenic defects, correct male infertility, and generate transgenic animals. PMID- 12077401 TI - Unique chromatin remodeling and transcriptional regulation in spermatogenesis. AB - Most of our knowledge of transcriptional regulation comes from studies in somatic cells. However, increasing evidence reveals that gene regulation mechanisms are different in haploid germ cells. A number of highly specialized strategies operate during spermatogenesis. These include a unique chromatin reorganization program and the use of distinct promoter elements and specific transcription factors. Deciphering the rules governing transcriptional control during spermatogenesis will provide valuable insights of biomedical importance. PMID- 12077402 TI - Intercellular communication in the mammalian ovary: oocytes carry the conversation. AB - The production of functional female gametes is essential for the propagation of all vertebrate species. The growth of oocytes within ovarian follicles and their development to mature eggs have fascinated biologists for centuries, and scientists have long realized the importance of the ovarian follicle's somatic cells in nurturing oogenesis and delivering the oocyte to the oviduct by ovulation. Recent studies have revealed key roles of the oocyte in folliculogenesis and established that bidirectional communication between the oocyte and companion somatic cells is essential for development of an egg competent to undergo fertilization and embryogenesis. The challenge for the future is to identify the factors that participate in this communication and their mechanisms of action. PMID- 12077403 TI - Sex matters in meiosis. AB - In mammals, fertilization typically involves the ovulation of one or a few eggs at one end of the female reproductive tract and the entry of millions of sperm at the other. Given this disparity in numbers, it might be expected that the more precious commodity-eggs-would be subject to more stringent quality-control mechanisms. However, information from engineered mutations of meiotic genes suggests just the opposite. Specifically, the available mutants demonstrate striking sexual dimorphism in response to meiotic disruption; for example, faced with adversity, male meiosis grinds to a halt, whereas female meiosis soldiers on. This female "robustness" comes with a cost, however, because aneuploidy appears to be increased in the resultant oocytes. PMID- 12077404 TI - Penetration, adhesion, and fusion in mammalian sperm-egg interaction. AB - Fertilization is the sum of the cellular mechanisms that pass the genome from one generation to the next and initiate development of a new organism. A typical, ovulated mammalian egg is enclosed by two layers: an outer layer of approximately 5000 cumulus cells and an inner, thick extracellular matrix, the zona pellucida. To reach the egg plasma membrane, sperm must penetrate both layers in steps requiring sperm motility, sperm surface enzymes, and probably sperm-secreted enzymes. Sperm also bind transiently to the egg zona pellucida and the egg plasma membrane and then fuse. Signaling in the sperm is induced by sperm adhesion to the zona pellucida, and signaling in the egg by gamete fusion. The gamete molecules and molecular interactions with essential roles in these events are gradually being discovered. PMID- 12077405 TI - Deciphering the cross-talk of implantation: advances and challenges. AB - Implantation involves a series of steps leading to an effective reciprocal signaling between the blastocyst and the uterus. Except for a restricted period when ovarian hormones induce a uterine receptive phase, the uterus is an unfavorable environment for blastocyst implantation. Because species-specific variations in implantation strategies exist, these differences preclude the formulation of a unifying theme for the molecular basis of this event. However, an increased understanding of mammalian implantation has been gained through the use of the mouse model. This review summarizes recognized signaling cascades and new research in mammalian implantation, based primarily on available genetic and molecular evidence from implantation studies in the mouse. Although the identification of new molecules associated with implantation in various species provides valuable insight, important questions remain regarding the common molecular mechanisms that govern this process. Understanding the mechanisms of implantation promises to help alleviate infertility, enhance fetal health, and improve contraceptive design. The success of any species depends on its reproductive efficiency. For sexual reproduction, an egg and sperm must overcome many obstacles to fuse and co-mingle their genetic material at fertilization. The zygote develops into a blastocyst with two cell lineages (the inner cell mass and the trophectoderm), migrates within the reproductive tract, and ultimately implants into a transiently permissive host tissue, the uterus. However, the molecular basis of the road map connecting the blastocyst with the endometrium across species is diverse (1) and not fully understood. Recent advances have identified numerous molecules involved in implantation (1-4), yet new discoveries have not yielded a unifying scheme for the mechanisms of implantation. PMID- 12077406 TI - The science of ART. AB - The methods of gamete manipulation used in assisted reproductive technology (ART) are rapidly proliferating and in some instances outpacing the underlying science. In this review, we discuss two major advances in the ART laboratory intracytoplasmic sperm injection and extended embryo culture before embryo transfer. We outline the rationale for these approaches, discuss results of experiments obtained from animal model systems and human preimplantation embryos that provide the scientific basis for these procedures, and point out potential concerns that have arisen from these studies. PMID- 12077407 TI - Amygdala response to happy faces as a function of extraversion. PMID- 12077409 TI - Coherent spin oscillations in a disordered magnet. AB - Most materials freeze when cooled to sufficiently low temperature. We find that magnetic dipoles randomly distributed in a solid matrix condense into a spin liquid with spectral properties on cooling that are the diametric opposite of those for conventional glasses. Measurements of the nonlinear magnetic dynamics in the low-temperature liquid reveal the presence of coherent spin oscillations composed of hundreds of spins with lifetimes of up to 10 seconds. These excitations can be labeled by frequency and manipulated by the magnetic fields from a loop of wire and can permit the encoding of information at multiple frequencies simultaneously. PMID- 12077408 TI - A continuous source of Bose-Einstein condensed atoms. AB - A continuous source of Bose-Einstein condensed sodium atoms was created by periodically replenishing a condensate held in an optical dipole trap with new condensates delivered using optical tweezers. The source contained more than 1 x 10(6) atoms at all times, raising the possibility of realizing a continuous atom laser. PMID- 12077410 TI - Antibody-based bio-nanotube membranes for enantiomeric drug separations. AB - Synthetic bio-nanotube membranes were developed and used to separate two enantiomers of a chiral drug. These membranes are based on alumina films that have cylindrical pores with monodisperse nanoscopic diameters (for example, 20 nanometers). Silica nanotubes were chemically synthesized within the pores of these films, and an antibody that selectively binds one of the enantiomers of the drug was attached to the inner walls of the silica nanotubes. These membranes selectively transport the enantiomer that specifically binds to the antibody, relative to the enantiomer that has lower affinity for the antibody. The solvent dimethyl sulfoxide was used to tune the antibody binding affinity. The enantiomeric selectivity coefficient increases as the inside diameter of the silica nanotubes decreases. PMID- 12077411 TI - Watching vibrational energy transfer in liquids with atomic spatial resolution. AB - Ultrafast spectroscopy was used to study vibrational energy transfer between vibrational reporter groups on different parts of a molecule in a liquid. When OH stretching vibrations of different alcohols were excited by mid-infrared laser pulses, vibrational energy was observed to move through intervening CH2 or CH groups, taking steps up and down in energy, ending up at terminal CH3 groups. For each additional CH2 group in the path between OH and CH3, the time for vibrational energy transfer increased by about 0.4 picosecond. PMID- 12077412 TI - Structure of the 8200-year cold event revealed by a speleothem trace element record. AB - Abrupt first-order shifts in strontium and phosphorus concentrations in stalagmite calcite deposited in western Ireland during the 8200-year event (the major cooling episode 8200 years before the present) are interpreted as responses to a drier climate lasting about 37 years. Both shifts are centered on 8330 +/- 80 years before the present, coinciding with a large oxygen isotope anomaly and a change in the calcite petrography. In this very high resolution (monthly) record, antipathetic second-order oscillations in phosphorus and strontium reveal decreased growth rates and increased rainfall seasonality. Growth rate variations within the event reveal a two-pronged structure consistent with recent model simulations. PMID- 12077413 TI - Insights into collisional magmatism from isotopic fingerprints of melting reactions. AB - Piston-cylinder experiments in the granite system demonstrate that a variety of isotopically distinct melts can arise from progressive melting of a single source. The relation between the isotopic composition of Sr and the stoichiometry of the observed melting reactions suggests that isotopic signatures of anatectic magmas can be used to infer melting reactions in natural systems. Our results also indicate that distinct episodes of dehydration and fluid-fluxed melting of a single, metapelitic source region may have contributed to the bimodal geochemistry of crustally derived leucogranites of the Himalayan orogen. PMID- 12077414 TI - A large paleolake basin at the head of Ma'adim Vallis, Mars. AB - At 8 to 15 kilometers wide, Ma'adim Vallis is one of the largest valleys in the martian highlands. Although a groundwater source was previously suggested, the channel originates at a spillway in the divide of a approximately 3,000,000 square-kilometer closed drainage basin. The interior morphology of this source basin, including likely shoreline features following topographic contours, suggests that Ma'adim Vallis was created through catastrophic overflow of a approximately 1,100,000-square-kilometer highland lake. The size, constant levels, and interior morphology of three regional paleolake basins require a warmer paleoclimate and a long-term, recharged, stable highland water table more than approximately 3.5 billion years ago. PMID- 12077415 TI - The mass disruption of Oort cloud comets. AB - We have calculated the number of dormant, nearly isotropic Oort cloud comets in the solar system by (i) combining orbital distribution models with statistical models of dormant comet discoveries by well-defined surveys and (ii) comparing the model results to observations of a population of dormant comets. Dynamical models that assume that comets are not destroyed predict that we should have discovered approximately 100 times more dormant nearly isotropic comets than are actually seen. Thus, as comets evolve inward from the Oort cloud, the majority of them must physically disrupt. PMID- 12077416 TI - Identification of signal peptide peptidase, a presenilin-type aspartic protease. AB - Signal peptide peptidase (SPP) catalyzes intramembrane proteolysis of some signal peptides after they have been cleaved from a preprotein. In humans, SPP activity is required to generate signal sequence-derived human lymphocyte antigen-E epitopes that are recognized by the immune system, and to process hepatitis C virus core protein. We have identified human SPP as a polytopic membrane protein with sequence motifs characteristic of the presenilin-type aspartic proteases. SPP and potential eukaryotic homologs may represent another family of aspartic proteases that promote intramembrane proteolysis to release biologically important peptides. PMID- 12077417 TI - Visualization and functional analysis of RNA-dependent RNA polymerase lattices. AB - Positive-strand RNA viruses such as poliovirus replicate their genomes on intracellular membranes of their eukaryotic hosts. Electron microscopy has revealed that purified poliovirus RNA-dependent RNA polymerase forms planar and tubular oligomeric arrays. The structural integrity of these arrays correlates with cooperative RNA binding and RNA elongation and is sensitive to mutations that disrupt intermolecular contacts predicted by the polymerase structure. Membranous vesicles isolated from poliovirus-infected cells contain structures consistent with the presence of two-dimensional polymerase arrays on their surfaces during infection. Therefore, host cytoplasmic membranes may function as physical foundations for two-dimensional polymerase arrays, conferring the advantages of surface catalysis to viral RNA replication. PMID- 12077418 TI - Pseudomonas-Candida interactions: an ecological role for virulence factors. AB - Bacterial-fungal interactions have great environmental, medical, and economic importance, yet few have been well characterized at the molecular level. Here, we describe a pathogenic interaction between Pseudomonas aeruginosa and Candida albicans, two opportunistic pathogens. P. aeruginosa forms a dense biofilm on C. albicans filaments and kills the fungus. In contrast, P. aeruginosa neither binds to nor kills yeast-form C. albicans. Several P. aeruginosa virulence factors that are important in disease are involved in the killing of C. albicans filaments. We propose that many virulence factors studied in the context of human infection may also have a role in bacterial-fungal interactions. PMID- 12077419 TI - Disassembly of transcriptional regulatory complexes by molecular chaperones. AB - Many biological processes are initiated by cooperative assembly of large multicomponent complexes; however, mechanisms for modulating or terminating the actions of these complexes are not well understood. For example, hormone-bound intracellular receptors (IRs) nucleate formation of transcriptional regulatory complexes whose actions cease promptly upon hormone withdrawal. Here, we show that the p23 molecular chaperone localizes in vivo to genomic response elements in a hormone-dependent manner, disrupting receptor-mediated transcriptional activation in vivo and in vitro; Hsp90 weakly displayed similar activities. Indeed, p23 and Hsp90 also disrupted the activities of some non-IR-containing transcriptional regulatory complexes. We suggest that molecular chaperones promote disassembly of transcriptional regulatory complexes, thus enabling regulatory machineries to detect and respond to signaling changes. PMID- 12077421 TI - Place cells and place recognition maintained by direct entorhinal-hippocampal circuitry. AB - Place cells in hippocampal area CA1 may receive positional information from the intrahippocampal associative network in area CA3 or directly from the entorhinal cortex. To determine whether direct entorhinal connections support spatial firing and spatial memory, we removed all input from areas CA3 to CA1, thus isolating the CA1 area. Pyramidal cells in the isolated CA1 area developed sharp and stable place fields. Rats with an isolated CA1 area showed normal acquisition of an associative hippocampal-dependent spatial recognition task. Spatial recall was impaired. These results suggest that the hippocampus contains two functionally separable memory circuits: The direct entorhinal-CA1 system is sufficient for recollection-based recognition memory, but recall depends on intact CA3-CA1 connectivity. PMID- 12077420 TI - Regulation of the different chromatin states of autosomes and X chromosomes in the germ line of C. elegans. AB - The Maternal-Effect Sterile (MES) proteins are essential for germline viability in Caenorhabditis elegans. Here, we report that MES-4, a SET-domain protein, binds to the autosomes but not to the X chromosomes. MES-2, MES-3, and MES-6 are required to exclude MES-4 and markers of active chromatin from the X chromosomes. These findings strengthen the emerging view that in the C. elegans germ line, the X chromosomes differ in chromatin state from the autosomes and are generally silenced. We propose that all four MES proteins participate in X-chromosome silencing, and that the role of MES-4 is to exclude repressors from the autosomes, thus enabling efficient repression of the Xs. PMID- 12077422 TI - Tech.Sight. Antibody design by man and nature. AB - Catalytic antibodies have emerged as being without peer as rationally designed biocatalysts. They have been shown to catalyze an ever-increasing array of chemical reactions with both high substrate specificity and selectivity. Probing the immune repertoire, via an expanding number of techniques, has lead to the production of proteins that can catalyze chemistry that is both difficult to perform using existing chemical methods and that is not catalyzed by endogenous enzymes. Remarkably, recent evidence has pointed to a hitherto unknown catalytic function of all antibodies that seems to be intrinsic to their immunoglobulin structure, the conversion of 1O2* into H2O2. This new catalytic potential of antibodies points to a new 'chemical arm' of the immune system and reveals that the evolution of catalytic antibodies significantly predates their rational design. PMID- 12077423 TI - Beaming light from a subwavelength aperture. AB - Light usually diffracts in all directions when it emerges from a subwavelength aperture, which puts a lower limit on the size of features that can be used in photonics. This limitation can be overcome by creating a periodic texture on the exit side of a single aperture in a metal film. The transmitted light emerges from the aperture as a beam with a small angular divergence (approximately +/-3 degrees ) whose directionality can be controlled. This finding is especially surprising, considering that the radiating region is mainly confined to an area with lateral dimensions comparable to the wavelength of the light. The device occupies no more than one cubic micrometer and, when combined with enhanced transmission, suggests that a wide range of photonic applications is possible. PMID- 12077424 TI - Generation and in vitro differentiation of a spermatogonial cell line. AB - Spermatogenesis is the process by which spermatogonial stem cells divide and differentiate to produce sperm. In vitro sperm production has been difficult to achieve because of the lack of a culture system to maintain viable spermatogonia for long periods of time. Here we report the in vitro generation of spermatocytes and spermatids from telomerase-immortalized mouse type A spermatogonial cells in the presence of stem cell factor. This differentiation can occur in the absence of supportive cells. The immortalized spermatogonial cell line may serve as a powerful tool in elucidating the molecular mechanisms of spermatogenesis. Furthermore, through genomic modification and transplantation techniques, this male germ cell line may be used to generate transgenic mice and to develop germ cell gene therapy. PMID- 12077426 TI - Ultrahigh-resolution structure of high-potential iron-sulfur protein from Thermochromatium tepidum. AB - Crystals of the high-potential iron-sulfur protein (HiPIP) from Thermochromatium tepidum diffract X-rays to 0.80 A using synchrotron radiation at 100 K. The crystal structure of this HiPIP was refined at this ultrahigh resolution with anisotropic temperature factors for all atoms to conventional crystallographic R factors of 0.092 and 0.101 for F(o) > 4sigma(F(o)) and all reflections, respectively. The present structure provides a more precise picture than the previous 1.5 A structure and allows location of the positions of most H atoms. The structure revealed a partly hydrophobic cavity near the main hydrophobic area and a much larger inter-cluster approach distance (23.454 A, the c constant of the unit cell) in the crystal packing than other types of HiPIPs. The structural features involved in the electron-transfer reaction of HiPIP are discussed. PMID- 12077425 TI - Dependence of heterochromatic histone H3 methylation patterns on the Arabidopsis gene DDM1. AB - The Arabidopsis gene DDM1 is required to maintain DNA methylation levels and is responsible for transposon and transgene silencing. However, rather than encoding a DNA methyltransferase, DDM1 has similarity to the SWI/SNF family of adenosine triphosphate-dependent chromatin remodeling genes, suggesting an indirect role in DNA methylation. Here we show that DDM1 is also required to maintain histone H3 methylation patterns. In wild-type heterochromatin, transposons and silent genes are associated with histone H3 methylated at lysine 9, whereas known genes are preferentially associated with methylated lysine 4. In ddm1 heterochromatin, DNA methylation is lost, and methylation of lysine 9 is largely replaced by methylation of lysine 4. Because DNA methylation has recently been shown to depend on histone H3 lysine 9 methylation, our results suggest that transposon methylation may be guided by histone H3 methylation in plant genomes. This would account for the epigenetic inheritance of hypomethylated DNA once histone H3 methylation patterns are altered. PMID- 12077427 TI - Generating isomorphous heavy-atom derivatives by a quick-soak method. Part I: test cases. AB - Screening for heavy-atom derivatives remains a time-consuming and cumbersome process that often results in non-isomorphous derivatives whose phases cannot be combined. Using lysozyme and FcgammaRIII receptor crystals as test cases, an improved soaking method for the generation of conventional heavy-atom derivatives has been developed. The method is based on soaking crystals in heavy-atom compounds for a very brief time at near-saturation concentrations. Compared with the current heavy-atom soaking method, which often takes days to achieve a derivatization, the quick-soak method completes a derivatization within 10 min to 2 h. The bound heavy-atom sites display higher peak heights from quick soaks than from overnight soaks in all cases tested. The quick-soak derivatives also preserved native-like diffraction resolution and data quality that was better than the prolonged-soak derivatives. Furthermore, derivatives generated by brief soaks are more isomorphous to the native than those generated by overnight soaks. Short soaks not only increase the likelihood of success in heavy-atom screening by reducing the pitfalls associated with prolonged soaks, such as lack of isomorphism and overall lattice disorder, but also have the potential to transform a time-consuming derivative screening into an 'on-the-fly' real-time derivatization process. PMID- 12077429 TI - Structure of cytochrome c6 from Arthrospira maxima: an assembly of 24 subunits in a nearly symmetric shell. AB - Cytochrome c(6) from the cyanobacterium Arthrospira maxima is present in isoforms that can be resolved by size-exclusion chromatography. One isoform crystallized in space group I4(1)32 with eight protein molecules in the asymmetric unit and a total of 384 molecules in the unit cell. Within the crystal, the molecules are arranged as clusters of 24 cytochrome c(6) molecules. Each cluster is a hollow shell with approximate octahedral (432) symmetry. Structural and biochemical studies of cytochrome c(6) isolated from other cyanobacteria and algae have led to the suggestion that cytochrome c(6) forms oligomers. The cytochrome c(6) complex described here is the largest assembly of cytochrome c(6) molecules observed thus far. PMID- 12077428 TI - Generating isomorphous heavy-atom derivatives by a quick-soak method. Part II: phasing of new structures. AB - A quick-soak method has been applied to generate de novo heavy-atom phasing to solve two new protein structures, a type II transforming growth factor beta receptor (TBRII) and a natural killer cell receptor-ligand complex, NKG2D-ULBP3. In the case of TBRII, a crystal derivatized for only 10 min in saturated HgCl(2) provided adequate phasing for structure determination. Comparison between HgCl(2) derivatives generated by 10 min soaking and by 12 h soaking revealed similar phasing statistics. The shorter soak, however, resulted in a derivative more isomorphous to the native than the longer soak as judged by changes in the unit cell parameter a upon derivatization as well as by the quality of a combined SIRAS electron-density map. In the case of the NKG2D-ULBP3 structure, all overnight soaks in heavy-atom solutions resulted in crystal lattice disorder and only the quick soaks preserved diffraction. Despite fragile lattice packing, the quick-soaked K(2)PtCl(4) derivative was isomorphous with the native crystal and the electron-density map calculated from combined SIR and MAD phases is better than that calculated from MAD phases alone. Combined with mass-spectrometry assisted solution heavy-atom derivative screening and the use of synchrotron radiation, the quick-soak derivatization has the potential to transform the time consuming conventional heavy-atom search into a real-time 'on-the-fly' derivatization process that will benefit high-throughput structural genomics. PMID- 12077430 TI - Thiol-reactive lanthanide chelates for phasing protein X-ray diffraction data. AB - Lanthanides can contribute a large anomalous component to X-ray scattering when present and ordered in a target crystal. This large anomalous signal is a useful source of phase information in X-ray crystallographic studies of biological macromolecules. Thiol-reactive lanthanide chelates were tested as a means of incorporation of lanthanides into protein crystals. Two compounds, each capable of being loaded with a lanthanide of choice, were synthesized: diethylenetriaminepentaacetic 3-(2-pyridyldithio)propionyl hydrazide (DTPA-PDPH) and 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic 3-(2 pyridyldithio)propionyl hydrazide (DOTA-PDPH). A cysteine mutant of the 34 kDa phosphate-binding protein (PBP-A197C) from Escherichia coli was used as a test case. PBP-A197C was labeled with DTPA-PDPH loaded with dysprosium. Characteristics of DTPA-PDPH enabled spectroscopic monitoring of the labeling reaction. Complete labeling of PBP-A197C was confirmed by mass spectrometry and SDS-PAGE analysis. Labeled PBP-A197C (PBP-A197C-DTPA-Dy) crystallized identically to unlabeled protein. X-ray diffraction data were collected from PBP-A197C-DTPA Dy crystals in-house with a Cu Kalpha rotating-anode source and with a tuneable synchrotron source (ALS 5.0.2). Synchrotron data were collected at energies corresponding to the Dy L(III) edge f" peak and a high-energy remote. Each data set was treated as an independent SAD experiment. A large anomalous signal was present in the data collected in-house and at the synchrotron. The Dy site was easily located in anomalous difference Patterson maps calculated from each of the data sets. In each case, SAD phasing resulted in high-quality electron-density maps, as evidenced by the success of automated model building. The generality of the method was analyzed with several other test proteins. Labeling of some of these proteins with thiol-reactive lanthanide chelates was deleterious to protein solubility or crystallization. In two of the cases the lanthanide chelate was disordered in the crystals. These results suggest that this method may not be well suited for high-throughput crystallography. However, for difficult cases requiring a large anomalous signal, thiol-reactive lanthanide chelates may prove to be a valuable tool. PMID- 12077431 TI - The 1.35 A structure of cadmium-substituted TM-3, a snake-venom metalloproteinase from Taiwan habu: elucidation of a TNFalpha-converting enzyme-like active-site structure with a distorted octahedral geometry of cadmium. AB - The crystal structure of TM-3, a small snake-venom metalloproteinase (SVMP) isolated from Taiwan habu (Trimeresurus mucrosquamatus), was determined at 1.35 A resolution with resultant R and R(free) values of 0.181 and 0.204, respectively. The overall structure of TM-3 is an oblate ellipsoid that contains three disulfide crosslinks, Cys118-Cys197, Cys159-Cys181 and Cys161-Cys164. It exhibits the typical structural features of SVMPs and is closely related to the structure of the catalytic proteinase domain of TNFalpha-converting enzyme (TACE). In the present structure, the essential catalytic zinc ion was found to be replaced by a cadmium ion during crystallization, as revealed by atomic absorption analysis and X-ray data. This cadmium ion is bound to six ligands, including three conserved histidines and three water molecules, displaying the coordination geometry of a distorted octahedron. One of the water molecules is proposed to play the role of stabilizing the tetrahedral intermediate during the catalysis of SVMPs. The putative S'(1) specificity pocket of TM-3 is relatively shallow, in contrast to the deep pockets of adamalysin II, atrolysin C and H(2)-proteinase, but is similar to those in acutolysin A and TACE. The shallow pocket is a consequence of the presence of the non-conserved disulfide bond Cys159-Cys181 and the residue Gln174 at the bottom of the S'(1) pocket. The results indicate that the active site structure of TM-3, among the know structures of SVMPs examined thus far, is most similar to that of TACE owing to their close disulfide configurations and the S'(1) specificity pocket. PMID- 12077432 TI - An enzyme with a deep trefoil knot for the active-site architecture. AB - Knots in polypeptide chains have been found in very few proteins. Only two proteins are considered to have a shallow 'trefoil' knot, which tucks a few residues at one end of the chain through a loop exposed on the protein surface. Recently, another protein was found by a mathematical algorithm to have a deep 'figure-of-eight' knot which had not been visually identified. In the present study, the crystal structure of a hypothetical RNA 2'-O-ribose methyltransferase from Thermus thermophilus (RrmA) was determined at 2.4 A resolution and a deep trefoil knot was found for the first time. The present knot is formed by the threading of a 44-residue polypeptide chain through a 41-residue loop and is better defined than the previously reported knots. Two of the three catalytic residues conserved in the 2'-O-ribose methyltransferase family are located in the knotting loop and in the knotted carboxy-terminal chain, which is the first observation that the enzyme active site is constructed right on the knot. On the other hand, the amino-terminal domain exhibits a geometrical similarity to the ribosomal proteins which recognize an internal loop of RNA. PMID- 12077433 TI - NH3-dependent NAD+ synthetase from Bacillus subtilis at 1 A resolution. AB - The final step of NAD+ biosynthesis includes an amide transfer to nicotinic acid adenine dinucleotide (NaAD) catalyzed by NAD+ synthetase. This enzyme was co crystallized in microgravity with natural substrates NaAD and ATP at pH 8.5. The crystal was exposed to ammonium ions, synchrotron diffraction data were collected and the atomic model was refined anisotropically at 1 A resolution to R = 11.63%. Both binding sites are occupied by the NAD-adenylate intermediate, pyrophosphate and two magnesium ions. The atomic resolution of the structure allows better definition of non-planar peptide groups, reveals a low mean anisotropy of protein and substrate atoms and indicates the H-atom positions of the phosphoester group of the reaction intermediate. The phosphoester group is protonated at the carbonyl O atom O7N, suggesting a carbenium-ion structure stabilized by interactions with two solvent sites presumably occupied by ammonia and a water molecule. A mechanism is proposed for the second catalytic step, which includes a nucleophilic attack by the ammonia molecule on the intermediate. PMID- 12077434 TI - Ab initio crystallographic structure determination of insulin from protein to electron density without crystal handling. AB - Insulin crystals suitable for cryogenic data collection and structure determination by single-wavelength anomalous scattering (SAS) were obtained by a self-optimization screening process in a single capillary tube without manipulation of the crystals at any time. Using the counter-diffusion crystallization technique, screening for optimal conditions for crystal growth, incorporation of a strong anomalous scattering halide and cryogenic solution took place simultaneously in a single capillary tube. The crystals in the capillaries can be placed directly in the cryostream for data collection using a conventional home-laboratory X-ray source. High-redundancy data were used to obtain a Patterson solution from the anomalous signal of iodine. As a result, the anomalous scattering-atom position was determined and the phase calculated, giving rise to an initial electron-density map at 2.4 A resolution. This entire procedure from crystal growth to the determination of an initial structure was performed within four weeks. PMID- 12077435 TI - Structures of human oxy- and deoxyhaemoglobin at different levels of humidity: variability in the T state. AB - High-salt crystals of human oxy- and deoxyhaemoglobin have been studied at different levels of environmental humidity and solvent content. The structure of the oxy form remains relatively unchanged at all levels. The deoxy form, however, undergoes a water-mediated transformation when the relative humidity around the crystals is reduced below 93%. The space group is maintained during the transformation, but the unit-cell volume nearly doubles, with two tetrameric molecules in the asymmetric unit of the low-humidity form compared with one in the native crystals. Interestingly, the haem geometry in the low-humidity form is closer to that in the oxy form than to that in the native deoxy form. The quaternary structure of one of the tetramers moves slightly towards that in the oxy form, while that in the other is more different from the oxy form than that in the high-salt native deoxy form. Thus, it would appear that, as in the case of the liganded form, the deoxy form of haemoglobin can also access an ensemble of related T states. PMID- 12077436 TI - Effect of stabilizing additives on the structure and hydration of proteins: a study involving monoclinic lysozyme. AB - In pursuance of a long-range programme on the hydration, mobility and action of proteins, the structural basis of the stabilizing effect of sugars and polyols is being investigated. With two crystallographically independent molecules with slightly different packing environments in the crystal, monoclinic lysozyme constitutes an ideal system for exploring the problem. The differences in the structure and hydration of the two molecules provide a framework for examining the changes caused by stabilizing additives. Monoclinic crystals were grown under native conditions and also in the presence of 10% sucrose, 15% trehalose, 10% trehalose, 10% sorbitol and 5% glycerol. The crystal structures were refined at resolutions ranging from 1.8 to 2.1 A. The average B values, and hence the mobility of the structure, are lower in the presence of additives than in the native crystals. However, a comparison of the structures indicates that the effect of the additives on the structure and the hydration shell around the protein molecule is considerably less than that caused by differences in packing. It is also less than that caused by the replacement of NaNO(3) by NaCl as the precipitant in the crystallization experiments. This result is not in conformity with the commonly held belief that additives exert their stabilizing effect through the reorganization of the hydration shell, at least as far as the ordered water molecules are concerned. PMID- 12077437 TI - Alternate conformations observed in catalytic serine of Bacillus subtilis lipase determined at 1.3 A resolution. AB - Bacillus subtilis extracellular lipase (BsL) has an exceptionally low molecular weight (19.4 kDa) for a member of the lipase family. A crystallographic study was performed on BsL in order to design and produce mutant BsL that will be more suitable for industrial uses based on analysis of the three-dimensional structure. Recently, the crystal structure of BsL has been determined at 1.5 A resolution [van Pouderoyen et al. (2001). J. Mol. Biol. 309, 215-226]. In the present study, a new crystal form of BsL which provides diffraction data to higher resolution was obtained and its structure was determined at 1.3 A using the MAD method. It was found that the active-site residue Ser77 has alternate side-chain conformations. The O(gamma) atom of the first conformer forms a hydrogen bond to the N(epsilon) atom of His155, a member of the catalytic triad. In contrast, the second conformer is constructed with a hydrogen bond to the side chain atom of the adjacent His76. These two conformers presumably correspond to the active and inactive states, respectively. Similar alternate conformations in the catalytic serine residue have been observed in Fusarium solani cutinase determined at 1.0 A resolution and Penicillium purpurogenum acetylxylan esterase at 0.9 A resolution. In addition, a glycerol molecule, which was used as a cryoprotectant, is found to be located in the active site. On the basis of these results, a model for substrate binding in the reaction-intermediate state of BsL is proposed. PMID- 12077438 TI - Structure of the functional domain of the major grass-pollen allergen Phlp 5b. AB - The major allergen Phlp 5b from timothy grass pollen induces allergic rhinitis and bronchial asthma in millions of allergic patients worldwide. As an important step towards understanding the interactions between the pollen protein and components of the human immune system, the structure of the C-terminal key domain of Phlp 5b has been determined at 2.0 A resolution and refined to an R value of 19.7%. This is the first known allergen composed entirely of alpha-helices. The protein forms a dimer stabilized by one intermolecular disulfide bridge. Sequence homology suggests that at least all group V and group VI grass-pollen allergens belong to this new class of 'four-helix-bundle allergens'. PMID- 12077439 TI - Structure of the C-terminally truncated human ProMMP9, a gelatin-binding matrix metalloproteinase. AB - The X-ray crystal structure of the proform of human matrix metalloproteinase MMP9 has been solved to 2.5 A resolution. The construct includes the prodomain, the catalytic domain and three FnII (fibronectin type II) domains. The prodomain is inserted into the active-site cleft, blocking access to the catalytic zinc. Comparison with the crystal structure of the most closely related MMP, MMP2, indicates that the conformations of residues in the active-site cleft and in the cysteine-switch peptide of the prodomain are highly conserved and that design of MMP9-specific inhibitors will be challenging. In common with MMP2, the MMP9 S1' inhibitor-binding pocket is large compared with that of other MMPs. One small point of difference in the S1' binding pockets of MMP9 and MMP2 may provide an opportunity to explore the design of specific inhibitors. The side chain of Arg424 in MMP9 is angled slightly away from the S1' pocket when compared with the corresponding residue in MMP2, Thr424. The secondary structure of the FnII domains is conserved between the two closely related MMPs, although the second FnII domain makes no contact with the catalytic domain in MMP9, while the same domain in MMP2 has a substantial area of interaction with the catalytic domain. PMID- 12077440 TI - Crystallization and preliminary X-ray crystallographic studies on recombinant human carnitine acetyltransferase. AB - In this paper, the purification, crystallization and preliminary X-ray crystallographic studies of human carnitine acetyltransferase are reported. Recombinant human carnitine acetyltransferase crystals were grown by the hanging drop vapor-diffusion method and belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 137.65, b = 84.76, c = 57.65 A and one molecule per asymmetric unit. The intensity data were collected from a cryocooled crystal to 1.6 A resolution using a conventional X-ray source. PMID- 12077441 TI - Crystallization and preliminary X-ray crystallographic studies of HLA-A*1101 complexed with an HIV-1 decapeptide. AB - A major goal of vaccine research for the prevention of AIDS is to determine the immune correlates of protection against HIV-1 infection. In this context, it is of interest to understand how HLA-A*1101, a significantly more prevalent class I allele in a cohort of highly HIV-1-exposed persistently seronegative individuals, functions in relation to protective immunity to HIV-1. Towards this goal, a soluble recombinant HLA-A*1101 molecule has been expressed and used to assemble a complex with beta2-microglobulin and a Nef decapeptide. The HLA-A*1101/beta2m/Nef complex was crystallized by the hanging-drop vapor-diffusion method. The crystal formed in the monoclinic space group P2(1), with unit-cell parameters a = 77.2, b = 88.5, c = 64.8 A, beta = 90.1 degrees, and contains two molecules in the asymmetric unit. A data set to 2.2 A resolution was collected and structure determination by molecular replacement is currently in progress. Understanding the three-dimensional structure of the HLA-A*1101/beta2m/Nef complex may provide insight into the functional role of this class I allele in relation to protective immunity to HIV-1. PMID- 12077442 TI - Crystallization of a stringent response factor from Aquifex aeolicus. AB - The crystallization of a key enzyme from Aquifex aeolicus with suggested bifunctional activity, acting as an exopolyphosphatase and a guanosine pentaphosphate phosphohydrolase, is reported. Native data were collected to below 2 A resolution from an orthorhombic crystal with unit-cell parameters a = 50.8, b = 70.3, c = 90.9 A. Methionine residues were introduced by mutation and deliberate oxidation of the protein allowed us to produce additional crystal forms with reproducible diffraction ability and increased phasing potential. This is the first report on the crystallization of a member of the Ppx/GppA phosphatase family. PMID- 12077443 TI - Purification and initial crystallization studies of a DnaB intein from Synechocystis sp. PCC 6803. AB - A 154-residue mini-intein from the dnaB gene of Synechocystis sp. PCC 6803 (Ssp DnaB intein) has been purified and crystallized using PEG 4000 as a precipitant. The crystal belongs to space group P3(1)21 or P3(2)21, with unit-cell parameters a = b = 58.2, c = 70.3 A. It has one molecule per asymmetric unit and diffracts to beyond 2.0 A under cryoconditions (100 K) using a rotating copper anode X-ray generator. PMID- 12077444 TI - Crystallographic study of a naturally occurring trans-splicing intein from Synechocystis sp. PCC 6803. AB - A naturally occurring split intein from the dnaE gene of Synechocystis sp. PCC6803 (Ssp DnaE intein) has been purified and crystallized using PEG 8K as precipitant. The crystal belongs to space group P3(1)21 or P3(2)21, with unit cell parameters a = b = 58.5, c = 70.2 A. It has one molecule per asymmetric unit and diffracts to beyond 2.9 A under cryoconditions (110 K) using a Cu rotating anode X-ray generator in-house. PMID- 12077445 TI - Crystallization and preliminary X-ray analysis of the Escherichia coli adaptor protein ClpS, free and in complex with the N-terminal domain of ClpA. AB - Protein degradation in Escherichia coli is accomplished by a handful of large oligomeric complexes. In most cases, these proteolytic machines are comprised of a chaperone (e.g. ClpA) that is required to prepare the substrate for degradation by the peptidase (e.g. ClpP). Recently, it was shown that the substrate recognition of the chaperone ClpA could be modified by the adaptor protein ClpS. To investigate the structural implications of this change in substrate specificity, ClpS was crystallized alone and in complex with the N-terminal domain of ClpA (ClpA(N)). Crystals of ClpS diffract to 2.9 A resolution and belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 82.63, b = 145.67, c = 152.31 A. Two different crystal forms of the ClpA(N)-ClpS complex were characterized. Crystal form I (CFI) belongs to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 91.63, b = 112.47, c = 38.47 A; data to 1.92 A resolution were collected. Crystals of form II (CFII) belong to space group P4(1/3)2(1)2, with unit-cell parameters a = b = 93.57, c = 78.77 A, and diffract to 1.85 A resolution. Data sets collected from heavy-atom derivatives of CFI indicated the incorporation of Pt and Hg atoms. Structure solution using MIR and MAD methods is currently under way. PMID- 12077446 TI - Expression, purification, crystallization and preliminary X-ray analysis of rat ecto-ADP-ribosyltransferase 2 (ART2.2). AB - ADP-ribosyltransferases catalyze the transfer of the ADP-ribose moiety from NAD(+) onto proteins and other targets. These enzymes have been found in prokaryotes and in vertebrates; a eukaryotic enzyme structure is not yet known. The enzyme from Rattus norvegicus was expressed in the Escherichia coli periplasm at a level of about 0.2 mg per litre of culture, purified and crystallized. Native data sets were collected to 2.0 A resolution. A self-rotation function revealed a local twofold axis in crystal form A and a Patterson function showed a translational relationship in form B. Form C contains only one molecule in the asymmetric unit. PMID- 12077447 TI - Crystallization and preliminary crystallographic studies of human TGF-beta type II receptor ligand-binding domain. AB - Three constructs (residues 15-136, 22-136 and 27-136) of the truncated extracellular domain of human transforming growth factor beta type II receptor (TBRII) were overexpressed in Escherichia coli. The constructs are referred to as TBRII(15-136), TBRII(22-136) and TBRII(27-136). The refolded receptors were purified using a combination of ion-exchange and size-exclusion chromatography. The purified receptors have an apparent molecular weight of 14 kDa as judged by size-exclusion chromatography. In the crystallization trials, TBRII(15-136) and TBRII(22-136) formed mostly crystal-like spheres but failed to produce data quality crystals. TBRII(27-136) yielded large single crystals from hanging drops using the vapor-diffusion procedure with PEG 2000 or 4000 at pH 5.0. The crystals diffracted to 1.05 A [using the X9B beamline operated at lambda = 1.0092 A of the National Synchrotron Light Source (NSLS) at the Brookhaven National Laboratory] and belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 35.5, b = 40.7, c = 76.2 A. There was one molecule in the asymmetric unit, which corresponds to a solvent content of 42.1%. PMID- 12077448 TI - Crystallization and preliminary X-ray crystallographic analysis of a yedU gene product from Escherichia coli. AB - A yedU gene product with a molecular mass of 31 kDa is a hypothetical protein with no known function. The protein was purified and crystallized at 296 K. X-ray diffraction data have been collected to 2.3 A using synchrotron radiation. The crystals belong to the primitive orthorhombic system, with unit-cell parameters a = 50.56, b = 63.45, c = 168.02 A. The asymmetric unit contains two monomers of the protein, with a corresponding V(M) of 2.25 A(3) Da(-1) and a solvent content of 44.84%. PMID- 12077449 TI - Crystallization of a nickel-containing superoxide dismutase and preliminary phase determination by MAD at the Ni K edge. AB - Superoxide dismutases are metalloenzymes which catalyse the disproportion of superoxide radicals and thus play an important role in the protection of biomolecules from oxidative damage. Redox-active metal ions known to act as the catalytic centre of these enzymes are Cu, Mn or Fe. Recently, enzymes containing Ni have been found in various Streptomyces species, introducing a fourth type of metal ion to the superoxide dismutase family. NiSOD has been crystallized for the purpose of structure determination by X-ray crystallography using Ni as an anomalous scatterer in multiple-wavelength anomalous dispersion (MAD) experiments. Two crystal forms belonging to space group P2(1)2(1)2(1) and one belonging to space group R3 were obtained using ammonium sulfate as a precipitant. Patterson maps of one of the orthorhombic forms revealed the presence of pseudo-translation, which could be removed for the other orthorhombic form by using 10% glycerol in its crystallization conditions. In addition, this reduced the unit cell by half. Phase information which led to interpretable electron-density maps was derived from MAD data to 2.0 A resolution after density modification applying solvent flattening, histogram matching and NCS averaging. Phases were extended to 1.68 A resolution with a data set collected at a wavelength of 1 A. Model building based on the resulting electron-density maps is in progress. PMID- 12077450 TI - Preliminary crystallographic analysis of the cysteine desulfurase IscS from Escherichia coli. AB - IscS is a widely distributed cysteine desulfurase that catalyzes the pyridoxal phosphate dependent beta-elimination of sulfur from L-cysteine and plays a central role in the delivery of sulfur to a variety of metabolic pathways. Crystals of Escherichia coli IscS have been obtained by the hanging-drop vapor diffusion method using polyethylene glycol (PEG) as a precipitant. Initial seed crystals were obtained using PEG 6000 and sodium acetate, and diffraction-quality crystals were grown using a mixture of PEG 2000 and PEG 10 000 in the presence of sodium citrate. A complete native X-ray diffraction data set was collected from a single crystal at 103 K to a resolution of 2.1 A. The crystals belong to space group P2(1)2(1)2(1) and have unit-cell parameters a = 73.7086, b = 101.9741, c = 108.617 A (alpha = beta = gamma = 90 degrees ). Analysis of the Matthews equation and self-rotation function suggest two molecules per asymmetric unit, consistent with the presence of a single dimeric molecule. PMID- 12077451 TI - Purification, crystallization and preliminary structural studies of dTDP-4-keto-6 deoxy-glucose-5-epimerase (EvaD) from Amycolatopsis orientalis, the fourth enzyme in the dTDP-L-epivancosamine biosynthetic pathway. AB - The vancomycin class of antibiotics is regarded as the last line of defence against Gram-positive bacteria. The compounds used clinically are very complex organic molecules and are made by fermentation. The biosynthesis of these is complex and fascinating. Its study holds out the prospect of utilizing genetic engineering of the enzymes in the pathway in order to produce novel vancomycin analogues. In part, this requires detailed structural insight into substrate specificity as well as the enzyme mechanism. The crystallization of one of the enzymes in the chloroeremomycin biosynthetic pathway (a member of the vancomycin family), dTDP-3-amino-4-keto 2,3,6-trideoxy-3-C-methyl-glucose-5-epimerase (EvaD) from Amycolatopsis orientalis, is reported here. The protein is fourth in the pathway which makes a carbohydrate essential for the activity of chloroeremomycin. The crystals of EvaD diffract to 1.5 A and have unit-cell parameters a = 98.6, b = 72.0, c = 57.1 A with space group P2(1)2(1)2. Data to this resolution were collected at the European Synchrotron Radiation Facility. PMID- 12077452 TI - Crystallization and preliminary X-ray diffraction studies of the water-soluble state of the pore-forming toxin sticholysin II from the sea anemone Stichodactyla helianthus. AB - Sticholysin II (StnII) is a potent cytolytic protein produced by the sea anemone Stichodactyla helianthus. StnII belongs to the actinoporin family, a group of proteins which are characterized by their ability to spontaneously interact with biological membranes. The cytolytic character of these proteins is currently explained in terms of a molecular mechanism involving the formation of transmembrane pores. StnII has been crystallized using the hanging-drop vapour diffusion method at 291 K. Diffraction-quality crystals have unit-cell parameters a = 32.30, b = 119.73, c = 43.42 A, beta = 90.04 degrees and belong to the monoclinic space group P2(1). Diffraction data to a resolution of 1.71 A were collected at synchrotron facilities. PMID- 12077453 TI - Crystallization and preliminary X-ray crystallographic studies of monoacylglycerol lipase of the moderately thermophilic Bacillus sp. H-257. AB - Thermostable monoacylglycerol lipase (MGLP; EC 3.1.1.23) from the moderately thermophilic Bacillus sp. H-257 has a unique substrate specificity. It hydrolyzes monoacylglycerols but does not hydrolyze di- or triacylglycerols. Crystals of the enzyme were obtained by the hanging-drop vapour-diffusion method using ammonium sulfate as a precipitant and benzamidine as an additive. The orthorhombic crystals belong to the space group P2(1)2(1)2(1), with unit-cell parameters a = 43.53, b = 100.82, c = 108.17 A. The crystals diffract to at least 2.3 A resolution and a native data set has been collected to 2.6 A resolution on a CCD detector using synchrotron radiation. PMID- 12077454 TI - Expression, purification, crystallization and preliminary diffraction analysis of RNase P protein from Thermotoga maritima. AB - Ribonuclease P (RNase P), the ubiquitous endonuclease that catalyzes maturation of the 5'-end of tRNA in bacteria, is a ribonucleoprotein particle composed of one large RNA and one small protein. Two major structural types of bacterial RNase P RNA have been identified by phylogenetic comparative analysis: the A (ancestral) and B (Bacillus) types. The RNase P protein from Thermotoga maritima, a hyperthermophilic bacterium with an A-type RNase P RNA, has been expressed in Escherichia coli. A purification strategy was developed to obtain a protein preparation suitable for crystallization. Protein crystals suitable for diffraction studies were obtained and characterized. PMID- 12077455 TI - Crystallization of the FAD-independent acetolactate synthase of Klebsiella pneumoniae. AB - Leucine and valine are formed in a common pathway from pyruvate in which the first intermediate is 2-acetolactate. In some bacteria, this compound also has a catabolic fate as the starting point for the butanediol fermentation. The enzyme (EC 4.1.3.18) that forms 2-acetolactate is known as either acetohydroxyacid synthase (AHAS) or acetolactate synthase (ALS), with the latter name preferred for the catabolic enzyme. A significant difference between AHAS and ALS is that the former requires FAD for catalytic activity, although the reason for this requirement is not well understood. Both enzymes require the cofactor thiamine diphosphate. Here, the crystallization and preliminary X-ray diffraction analysis of the Klebsiella pneumoniae ALS is reported. Data to 2.6 A resolution have been collected at 100 K using a rotating-anode generator and an R-AXIS IV++ detector. Crystals have unit-cell parameters a = 137.4, b = 143.9, c = 134.4 A, alpha = 90, beta = 108.4, gamma = 90 degrees and belong to space group C2. Preliminary analysis indicates that there are four monomers located in each asymmetric unit. PMID- 12077456 TI - Expression, purification, crystallization and preliminary X-ray analysis of a DNA binding protein from Methanococcus jannaschii. AB - A small DNA-binding protein of 87 amino-acid residues from the hyperthermophilic archaeon Methanococcus jannaschii (Mja10b) was cloned and overexpressed in Escherichia coli. The protein was crystallized and the crystals belong to the space group P6(1)22/P6(5)22, with unit-cell parameters a = b = 50.85, c = 124.02 A, alpha = beta = 90, gamma = 120 degrees. The crystals diffracted to a maximum resolution of 2.2 A at 100 K using Cu Kalpha radiation. The presence of one molecule per asymmetric unit gives a crystal volume per protein mass (V(M)) of 2.4 A(3) Da(-1) and a solvent content of 49% by volume. A full set of X-ray diffraction data was collected to 2.2 A from the native crystal. PMID- 12077457 TI - Crystallization and preliminary analysis of Escherichia coli YodA. AB - The Escherichia coli protein YodA was overexpressed, purified and crystallized in several crystal forms. The function of this protein is not known, although it has been identified under conditions of bacterial stress. Three of the four crystal forms were obtained in the presence of divalent cations (zinc, nickel and cadmium), suggesting that YodA may be a metal-binding protein. PMID- 12077458 TI - Crystallization and preliminary structural analysis of an antibody complex formed with PfMSP1-19, a malaria vaccine candidate. AB - The 11 kDa C-terminal fragment of the proteolyticly matured surface antigen, PfMSP1, from Plasmodium falciparum is a promising malaria vaccine candidate. The soluble recombinant form of this naturally occurring fragment has been crystallized as a complex with the Fab of a specific murine monoclonal antibody. The crystals belong to the space group P2(1), with unit-cell parameters a = 51.8, b = 213.5,c = 60.0 A, beta =101.0 degrees, and with Z = 4. Diffraction data have been measured to 2.9 A resolution and a preliminary model of the complex has been determined by molecular replacement. The epitope recognised by G17.12 is located on the N-terminal EGF-like domain of the antigen. PMID- 12077459 TI - Characterization and crystallization of DivK, an essential response regulator for cell division and differentiation in Caulobacter crescentus. AB - DivK is an essential response regulator involved in the complex signal transduction network required for cell division and cell differentiation in Caulobacter crescentus. Small-angle X-ray scattering analysis was valuable for obtaining single crystals of the DivK recombinant protein. These crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 37.2, b = 40.5, c = 67.1 A and diffract beyond 1.6 A on a synchrotron beamline. PMID- 12077461 TI - The Social Physique Anxiety Scale: an example of the potential consequence of negatively worded items in factorial validity studies. AB - Social physique anxiety (SPA) based on Hart, Leary, and Rejeski's (1989) Social Physique Anxiety Scale (SPAS) was originally conceptualized to be a unidimensional construct. Empirical evidence on the factorial validity of the SPAS has been contradictory, yielding both one- and two-factor models. The two factor model, which consists of separate factors associated with positively and negatively worded items, has stimulated an ongoing debate about the dimensionality and content of the SPAS. The present study employed confirmatory factor analysis (CFA) to examine whether the two-factor solution to the 12-item SPAS was substantively meaningful or a methodological artifact. Results of the CFAs, which were performed on responses from four different samples (Eklund, Kelley, and Wilson, 1997; Eklund, Mack, and Hart, 1996), supported the existence of a single substantive SPA factor underlying responses to the 12-item SPAS. There were, in addition, method effects associated with the negatively worded items that could be modeled to achieve good fit. Therefore, it was concluded that a single substantive factor and a non-substantive method effect primarily related to the negatively worded items best represented the 12-item SPAS. PMID- 12077460 TI - Anisotropic behaviour of the C-terminal Kunitz-type domain of the alpha3 chain of human type VI collagen at atomic resolution (0.9 A). AB - The C-terminal Kunitz-type domain from the alpha3 chain of human type VI collagen (C5), a single amino-acid residue chain with three disulfide bridges, was refined at 0.9 A resolution in a monoclinic form, space group P2(1) with one molecule per asymmetric unit, using data collected at cryogenic temperature (110 K). The average protein factor decreases from 21 A(2) at room temperature (RT) to 12 A(2) at cryotemperature (100 K, CT). The spatially close N- and C-termini remain highly disordered. The different structural motifs of C5 were analyzed in terms of rigid-body displacement (TLS analyses) and show dominant libration motion for the secondary structure. PMID- 12077462 TI - Moral and evaluative reasoning across the life-span. AB - In a longitudinal/cross sectional study of moral and evaluative reasoning, Armon interviewed 23 females and 19 males, ages ranging from 5 at the first test time (1977) to 86 at the 4th (1989) test-time. Rasch analysis of Armon's data demonstrated that Armon's and Kohlberg's measures tap a single underlying dimension of reasoning; that individual stages across five items measure the same levels of reasoning, and that development on all items progresses at about the same rate. Participants found it easier to apply already available reasoning structures to new areas than to reason at a new stage, implying that stage transition is step-like. PMID- 12077463 TI - Methodological issues in using the Rasch model to select cross culturally equivalent items in order to develop a Quality of Life index: the analysis of four WHOQOL-100 data sets (Argentina, France, Hong Kong, United Kingdom). PMID- 12077464 TI - Simultaneous measurement of reading growth, gender, and relative-age effects: many-faceted Rasch applied to CBM reading scores. AB - Reading growth, gender effects, relative-age effects, and reading probe difficulty for reading were simultaneously assessed on one linear scale. The reading measure chosen was the Curriculum Based Measurement (CBM), words-read correctly. A sample of 1619 students in Grades Two through Seven was employed. There is growth in reading within all grades but decreasing growth with increasing grade level. There are consistent gender differences favoring girls but the differences are equivalent to only one month's growth. There is no consistent evidence of a relative-age effect across grades. Two strategies for linking data across grades were compared and found to produce results consistent with each other and individual grade results. PMID- 12077465 TI - Equating and item banking with the Rasch model. AB - This article describes Rasch measurement procedures for equating multiple test forms or calibrating an item bank. The procedures entail (a) selecting an appropriate data collection design, (b) estimating parameters, (c) transforming the parameters from multiple forms to a common scale, and (d) evaluating the quality of the linkage between these forms. Data collection designs include (a) anchor tests, (b) single group, (c) single data set, and (d) equivalent groups. Estimation procedures may involve (a) separate or (b) simultaneous calibration of data from multiple forms. Transformation is typically accomplished using (a) estimation scaling, but may involve (b) parameter anchoring or (c) computing equating constants. Link quality is evaluated using four fit indices: (a) item within-link, (b) item-between-link, (c) link-within-bank, and (d) form-within bank. These procedures are illustrated using an anchor test design. PMID- 12077466 TI - Treatment of cutaneous T cell lymphoma. AB - Primary cutaneous T cell lymphomas encompass a wide variety of lymphomas that are characterized by a distinct clinical presentation. Advanced biological techniques have allowed a more precise classification in recent years. Stage-adapted therapy is at present the best approach to treat cutaneous T cell lymphoma. PMID- 12077467 TI - Dermal toxicity due to industrial chemicals. AB - The dermal toxicity due to industrial chemicals is well recognised. They may be the cause of skin irritation and skin sensitization. Some of the chemicals may result in dermal reactions such as changes in skinfold thickness, cellular degeneration and necrosis. The absorption through the skin may also cause penetration of chemicals into the organism causing systemic effects. PMID- 12077468 TI - Local skin lesions in the rat after subcutaneous deposition of capsaicin. AB - Capsaicin is used to investigate the role of peripheral sensory nerve fibers. In previous studies of rats treated by injection of capsaicin into the skin of the neck, 'spontaneous' lesions in the head and neck region were observed. In this study, the course of development over time, the regional distribution and the innervation of capsaicin-induced dermal lesions were assessed in young male Sprague-Dawley rats. In one experiment, capsaicin was administered subcutaneously by injection in the skin of the neck. In a second experiment, capsaicin was injected in the back by a long needle that tunneled under the skin and allowed the capsaicin to be deposited in the subcutaneous fat of the neck. The density and the distribution of dermal nerve fibers were investigated by immunohistochemistry, using antisera against a panneuronal marker, protein gene product 9.5 (PGP), and calcitonin gene-related peptide (CGRP). In the first experiment, rats developed lesions in the neck area 11 days after injection. In the second experiment, lesions appeared in the skin of the back and occasionally in the neck area 10 days after injection. Development of lesions in the afflicted areas was paralleled by local reduction in the density of CGRP-immunoreactive (IR) nerve fibers, 80% in the first experiment and 72% in the second. The number of PGP-IR fibers was likewise reduced, by 39 and 41%, respectively. The density of the CGRP-IR fibers in the wound area was the same as in the adjacent, nonlesioned skin. The healing of the capsaicin-induced lesions was slow compared with surgical wounds in control animals. The wounds healed with hypertrophic scars. The healing process in the skin of the back was associated with the proliferation of CGRP-IR fibers. The study shows cutaneous lesions to appear in the region of the subcutaneous deposition of capsaicin. A uniform depletion of capsaicin-sensitive nerve fibers in the area of deposition suggests that an additional factor is needed to induce lesions. Possibly, impaired nociception in the afflicted area results in more vigorous grooming behavior and this, in turn, in a local skin damage. PMID- 12077469 TI - Urea and sodium chloride in moisturisers for skin of the elderly--a comparative, double-blind, randomised study. AB - Urea has long been used to treat dry skin. In the present report, we compared two identical creams, with the exception that one contained both urea and sodium chloride and the other urea alone, in 23 healthy elderly subjects. Following a 2 week wash-out, their clinically non-eczematous, rough- or normal-appearing skin on the anterior proximal part of the lower legs was treated twice daily in a double-blind and randomised manner. We examined the treated areas by measuring transepidermal water loss, capacitance and electrical impedance. Our findings suggest that both moisturisers seem equally effective, at least concerning the ability to reverse impedance indices towards normal, an effect ascribed to changes in hydration of the stratum corneum. However, the relevance of the impedance parameters to the clinical picture is disputable and further studies of moisturisers in elderly subjects are needed. PMID- 12077470 TI - Genistein and daidzein stimulate hyaluronic acid production in transformed human keratinocyte culture and hairless mouse skin. AB - We examined the effects of the soy isoflavones genistein (Gen) and daidzein (Dai) on the production of hyaluronic acid (HA) in a transformed human keratinocyte culture and in hairless mouse skin following topical application for 2 weeks. Gen and Dai, but not the glycosides thereof, significantly enhanced the production of HA in vitro and in vivo. Histochemistry using an HA-binding protein revealed that topical Gen and estradiol raised both the density and intensity of HA staining, which was abundant in the murine dermis. It is suggested that Gen and Dai are not released from their respective glycosides in culture or murine skin. Moreover, topical Gen and Dai may prevent and improve the cutaneous alterations caused by the loss of HA in skin. PMID- 12077472 TI - Systemic organophosphate poisoning following the percutaneous injection of insecticide. Case report. AB - Organophosphates are the most common group of chemicals in the southern part of Turkey. Although organophosphate poisoning (OPP) may occur due to skin exposure or inhalation, severe poisoning is usually the result of ingestion to attempt suicide. Despite the fact that there have been a lot of experimental studies using intravenous or percutaneous injection of organophosphates, reports of human poisoning due to percutaneous injection are rare. The systemic signs of OPP have not been described in these reported patients. We report 2 cases having systemic signs of OPP due to percutaneous injection. In our first case, we noticed a 17 day muscle weakness and a 12-day muscarinic syndrome, which required prolonged atropinization. In the second patient, atropine infusion had to be continued for 2 days. Both cases also had severe swelling of the affected limb and wound infection. In conclusion, in cases of percutaneous injection of organophosphates systemic toxicity may develop in addition to local findings such as necrosis and abscesses. Close observation for evidence of systemic involvement is required, and the patient should be carefully monitored for secondary abscess formation and any delayed impairment of neurologic function. PMID- 12077471 TI - Percutaneous absorption studies of octamethylcyclotetrasiloxane using the human skin/nude mouse model. AB - Octamethylcyclotetrasiloxane (D(4)) has been used for more than 40 years in industrial applications and consumer products, including the personal care industry. D(4) possesses many properties suitable for personal care products, such as low surface tension, water repellency, and thermal and chemical stability. The skin is a major route of exposure to D(4 )for humans. The main objective of this study was to evaluate the percutaneous absorption of neat D(4) in human skin using the human skin/nude mouse model. This information is needed to aid in assessing potential risks associated with the intended use of D(4). To determine whether D(4) accumulates in adipose tissue of the skin, the distribution of D(4) in human skin layers following application of neat D(4) was also evaluated. In this study, a mean of 1.09 +/- 0.46% of the applied dose was absorbed by the animal under semioccluded conditions. Only about 0.02% of the applied dose remained in the skin after 24 h of exposure (or 72 h after application). The majority (94.59 +/- 12.28%) of the dose evaporated from the site. Excretion in the volatile trap (or expired volatiles) accounted for 42% of the radioactivity that was absorbed, while 49% were excreted in the urine and feces. Despite the lipophilic properties of D(4), a significant accumulation of D(4) in adipose tissue of the skin was not observed 24 h following application to the surface of the skin. The small amount of D(4) detected in the skin was distributed mainly in the epidermis (61%), with lower amounts in the dermis (29%) and subcutaneous adipose tissue (10%). Dermal absorption studies using human skin transplanted onto nude mice showed that this model could be successfully applied for in vivo percutaneous absorption studies of D(4), and presumably of other cyclic siloxanes. The fraction of the percutaneous dose of D(4) absorbed in this model was found to be consistent with results reported by others using different experimental approaches. PMID- 12077473 TI - Effects of oral acitretin on contrast sensitivity and tear film function: a prospective study. AB - The purpose of this prospective study was to investigate the ocular side effects of short-term therapy with oral acitretin (1 mg/kg/day) in 24 patients with severe and recalcitrant dermatoses. Apart from the routine ophthalmological examination, the following tests were performed: break-up time of tear film for the determination of its stability, Schirmer test for the assessment of lacrimal gland function, rose bengal staining for the detection of possible ocular surface damage and contrast sensitivity test for the evaluation of visual function. No statistically significant differences could be found between the pretreatment values of the assessed parameters and those obtained after 1 and 2 months of therapy. It seems reasonable, therefore, to suggest that ocular surface integrity and tear film and visual function are not affected by short-term oral acitretin administration. PMID- 12077475 TI - Differences in quantitative characteristics of intracranial pressure in hydrocephalic children treated surgically or conservatively. AB - This study reports the results of quantitative analysis of continuous intracranial pressure (ICP) recordings in 33 hydrocephalic children. The aim of the study was to compare the exact numbers of increases in ICP during sleep or the awake state in hydrocephalic children who were treated either surgically or conservatively. At the time of ICP monitoring, the ICP curves were assessed by the calculation of mean ICP and visual inspection for the detection of plateau waves. Quantitative analysis was performed with the software Sensometrics Pressure Analyser, which presented the ICP curve as a matrix of numbers of ICP elevations of different levels (20-40 mm Hg) and durations (0.5-20 min). In each case, the numbers of ICP elevations were standardized to 10 h of recording time, providing the opportunity for comparisons of ICP curves between individuals. Compared to the surgery group, there was a rather high number of ICP elevations of 20 mm Hg of various durations in the nonsurgery group, e.g. ICP elevations of 20 mm Hg lasting 10 min occurred in 13 of 19 children (68%) in the nonsurgery group. There was no apparent relationship between ICP and age or between the size of the cerebral ventricles and ICP. In children with hydrocephalus, the presentation of the ICP data as a matrix of ICP elevations of different levels and durations may enhance the informative value of continuous ICP monitoring, as compared to the calculation of mean ICP and visual detection of plateau waves. PMID- 12077474 TI - A model of pulsations in communicating hydrocephalus. AB - The traditional theory of communicating hydrocephalus has implicated the bulk flow component of CSF motion; that is, hydrocephalus is generally understood as an imbalance between CSF formation and absorption. The theory that the cause of communicating hydrocephalus is malabsorption of CSF at the arachnoid villi is not substantiated by experimental evidence or by physical reasoning. Flow-sensitive MRI has shown that nearly all CSF motion is pulsatile, and there is substantial evidence that hyperdynamic choroid plexus pulsations are necessary and sufficient for ventricular dilation in communicating hydrocephalus. We have developed a model of intracranial pulsations based on the analogy between the pulsatile motion of electrons in an electrical circuit and the pulsatile motion of blood and CSF in the cranium. Increased impedance to the flow of CSF pulsations in the subarachnoid space redistributes the flow of pulsations into the ventricular CSF and into the capillary and venous circulation. The salient features of communicating hydrocephalus, such as ventricular dilation, intracranial pressure waves, narrowing of the CSF-venous pressure gradient, diminished cerebral blood flow, elevated resistive index and malabsorption of CSF, emerge naturally from the model. We propose that communicating hydrocephalus is the result of a redistribution of CSF pulsations in the cranium. PMID- 12077476 TI - Discernment of adipose versus nervous tissue: a novel adjunct solution in lipomyelomeningocele surgery. AB - OBJECTIVE: To determine a solution capable of discerning adipose versus nervous tissue, to aid in surgical separation of the adipose tissue which appears to be visually indistinguishable from nervous tissue in lipomyelomeningoceles (LMMs). METHODS: The following solutes (in normal saline) were investigated, both at 25 and 37 degrees C: beta-carotene, vitamin D, vitamin E, lecithin, hydrogen peroxide, lipase, protease, hyaluronidase, partially purified collagenase, purified collagenase, trypsin, trypsin plus purified collagenase and non-solute containing saline (control). Each solution was applied to a pediatric lipoma to determine gross effects over a period of approximately 30 min. If a solution appeared to affect the adipose tissue grossly, studies of functional in vivo sensory evoked and spontaneous potentials using that particular solution were conducted upon sheep spinal cord, nerve roots, dura and peripheral nerve. Additionally, histological studies were conducted to determine the effect of that solution upon adipose tissue, spinal cord, myelin, dura and nerve roots. RESULTS: Of all solutions investigated, partially purified collagenase type 1 (T1C; Lot M0M4322, Code CLS-1, Worthington Biochemical Corporation, Lakewood, N.J., USA) at 37 degrees C was the most successful in grossly altering the consistency and appearance of adipose tissue. This change was more apparent over 20-30 min following application of the solution to the adipose tissue. Solutions not containing T1C did not show appreciable results; purified collagenase plus trypsin did not appear comparable or superior to T1C. No significant histological or functional change was noted when comparing the spinal cord, nerve rootlets, myelin, dura or peripheral nerve from the T1C-treated group versus normal (untreated) control groups. CONCLUSION: T1C appears to be a potentially effective solution for application during LMM surgery in the acute setting, and such use of an adjunct solution may significantly aid in the safe surgical resection of LMMs. Pending further research, this technique may be applied for other indications which require discernment or alteration of adipose versus nervous tissue. PMID- 12077477 TI - Neuronal vacuolation in the basal nucleus of meynert caused by fetal hydrocephalus. AB - The basal nucleus of Meynert, which provides the major source of cholinergic input to the cerebral cortex, was investigated in fetal hydrocephalic brains (n = 5) in comparison to controls (n = 5). To detect alterations, the neurons of the basal nucleus were immunostained with an antibody against the calcium-binding protein calbindin. In the basal nucleus of hydrocephalic brains, a moderate number of vacuolated neurons were observed. Such vacuolation was not seen in control brains. Vacuolation is interpreted as a degenerative change after injury of basal nucleus axons in cases of fetal hydrocephalus. The neuronal alterations in the basal nucleus may account at least in part for residual deficits observed after shunt placement. PMID- 12077478 TI - Congenital arachnoid cyst mimicking meningocele. AB - The subject of this report is a rare case of a 5-year-old girl who developed an arachnoid cyst with a bony and dural defect in the parietal convexity. She had no history of head trauma or infection. Surgical exploration revealed the bulging lesion to consist of cerebrospinal fluid-containing spongy subcutaneous tissue and to extend into the bony and dural defect. The arachnoid cyst cavity was found beneath the subcutaneous lesion and was not connected to the adjacent subarachnoid space. Histologically, the subcutaneous tissue contained a complex of sinusoidal channels formed by an abundance of migrating arachnoidal cells, thus mimicking meningocele. PMID- 12077479 TI - Kinetics of binding of [(3)H]glycine to transport proteins in channel catfish brain. AB - [(3)H]Glycine was observed to bind to channel catfish brain particles in a manner displaying saturation kinetics. The dissociation constant was calculated to be 7.38 +/- 2.11 microM. Though some binding occurred in the absence of Na(+) ions, the presence of such ions stimulated binding in a concentration-dependent manner. Similarly, chloride ions had a stimulatory effect on [(3)H]glycine binding. Several inhibitors of binding were identified, the most effective being beta alanine, pipecolic acid, 2,3-pyrazine dicarboxylic acid and 3,5-pyrazole dicarboxylic acid. Each is a structural analogue of glycine. Harmaline, a known inhibitor of Na(+) binding, also inhibited glycine binding. A previous study had shown the presence of a sodium-dependent, active uptake system for glycine in synaptosomes derived from catfish brain. The present results suggest that the binding of [(3)H]glycine was to a glycine transporter and that the amino acid functions as a neurotransmitter in this species. PMID- 12077480 TI - Entrainment of circadian activity rhythms in rats to melatonin administered at T cycles different from 24 hours. AB - Entrainment of running wheel activity in DD was studied in adult male Long Evans rats exposed to cycles of a constant dose of melatonin (MEL; 100 microg/h) infused subcutaneously. The period (T) of the MEL cycle was initially kept at 24 h until stable entrainment was established; T was then changed in a stepwise manner, and each new T value was maintained for at least 20 cycles. Entrainment by phase advance occurred near circadian time 12 (activity onset), and the range of entrainment was between 30 and 35 min. The negative phase angle difference between activity onset and MEL onset increased as T values approached the entrainment limit, whereas no change in the duration of daily activity periods was found. No difference was observed between pre- and posttreatment values of the endogenous circadian period; hence, no aftereffects were found for any T value. These results indicate that the functional properties of entrainment to MEL are similar to those of entrainment to light, suggesting that both zeitgebers share a common timing mechanism. PMID- 12077481 TI - Age-related changes in 24-hour rhythms of norepinephrine content and serotonin turnover in rat pineal gland: effect of melatonin treatment. AB - The 24-hour rhythms of pineal norepinephrine (NE) content and serotonin (5-HT) turnover [estimated from the ratio of 5-hydroxyindoleacetic acid (5-HIAA) to 5 HT] were studied in young (2 months) and aged (18-20 months) Wistar rats killed at 6 different time points throughout a 24-hour cycle. In the first study, significant changes dependent on the time of day were identified, with acrophases in the first half of the activity span for both parameters. Old rats showed significantly smaller mesor and amplitude of the 24-hour rhythm of pineal NE content. They also showed decreased amplitude of the pineal 5-HT turnover rhythm, in the absence of changes in mesor. In old rats, pineal 5-HT and 5-HIAA concentrations were 41-47% of those found in young rats. In a second study, young and old rats received daily intraperitoneal injections of melatonin (30 microg) or vehicle for 11 days at 19.00 h (i.e. 11 h after light on). Analyzed as a main factor in a factorial analysis of variance, both pineal NE content and 5-HT turnover decreased in old rats while pineal 5-HT turnover increased after melatonin treatment. Melatonin treatment augmented the amplitude of the 24-hour rhythm of pineal NE content by 120 and 52% in young and old rats, respectively. The amplitude of the 24-hour rhythm of pineal 5-HT turnover almost doubled after melatonin treatment in young rats and did not change in old rats. Melatonin injection did not modify the rhythm's acrophase. The results indicate that old rats had lower amplitude and lower mesor values of 24-hour variations in pineal NE content and 5-HT turnover. Melatonin treatment only partly restored pineal NE content and was devoid of activity on pineal 5-HT turnover and 5-HT and 5-HIAA concentration in old rats. Impairment of pineal melatonin synthesizing capacity and intrapineal responses to melatonin may underlie pineal aging in rats. PMID- 12077482 TI - Co-existence of protein kinase C gamma and calcium-binding proteins in neurons of the medullary dorsal horn of the rat. AB - Protein kinase C gamma isoform (PKCgamma) is present at high levels in the spinal and medullary dorsal horns and is thought to play a role in the sensitization of dorsal horn neurons in certain pain states. Calbindin-D28k (CB), calretinin (CR) and parvalbumin (PV) are the most commonly expressed calcium-binding proteins and are located abundantly in the medullary dorsal horn (also called the caudal subnucleus of the spinal trigeminal nucleus). In the present study, immunofluorescence histochemical double staining for PKCgamma and CB, CR or PV was performed in the rat medullary dorsal horn. Most of the PKCgamma-, CB-, CR- and PV-immunoreactive neurons were observed in lamina II; some were also encountered in lamina I and lamina III of the medullary dorsal horn. Neurons co expressing CB/PKCgamma, CR/PKCgamma and PV/PKCgamma were also mainly found in lamina II, while in lamina I and lamina III, only a few neurons co-expressing CB/PKCgamma, CR/PKCgamma and PV/PKCgamma were encountered. The percentages of neurons co-expressing CB/PKCgamma in the total numbers of CB- and PKCgamma immunoreactive neurons were 6.7 and 5.9%, respectively. Of the total numbers of CR- and PKCgamma-immunoreactive neurons, 5.0 and 5.6%, respectively, showed both CR and PKCgamma immunoreactivities. The percentages of neurons co-expressing PV/PKCgamma in the total numbers of PV- and PKCgamma-immunoreactive neurons were 25.7 and 4.1%, respectively. Most of these neurons co-expressing CB/PKCgamma, CR/PKCgamma and PV/PKCgamma were small (/=36 microm) multipolar neurons were infrequently seen. The present results indicate that there are some neurons co-expressing CB/PKCgamma, CR/PKCgamma and PV/PKCgamma in the medullary dorsal horn. These neurons might play important roles in the nociceptive modulation from the oro-facial region. PMID- 12077483 TI - Expression of a kinase anchoring protein 79 and synaptophysin in the developing human red nucleus. AB - Our previous study showed that in the human fetal and neonatal brain, the magnocellular and parvocellular parts of the red nucleus can be well delineated by calcium-binding proteins. To study the development of rubral afferents, the expression of A kinase anchoring protein 79 (AKAP79) and synaptophysin (SYN) was examined in the human fetal red nucleus. It was found that during prenatal development both AKAP79 and SYN expression increased gradually although a major alteration in the distribution of the proteins within the two compartments of the red nucleus was not observed. In AKAP79 immunopreparations, the magnocellular part became well demarcated from 23 weeks of gestation onwards and both parts showed punctate immunolabelling with moderate to high packing densities of immunoreactive cells. SYN immunoreactivity with a punctate appearance was, however, mainly located in the parvocellular part. It was evenly distributed throughout the compartment at 14-22 weeks of gestation, and then from 23 weeks to the time of birth, there was a pericellular arrangement of SYN. Our observations are mainly in line with connectivity data regarding the red nucleus. PMID- 12077484 TI - Regulation of period 1 expression in cultured rat pineal. AB - The aim of the present study was to investigate the in vitro expression of Period 1 (Per1), Period 2 (Per2) and arylalkylamine N-acetyltransferase (AA-NAT) genes in the rat pineal gland to understand the mechanism(s) regulating the expression of these genes in this organ. Pineals, when maintained in vitro for 5 days, did not show circadian rhythmicity in the expression of any of the three genes monitored. Norepinephrine (NE) induced AA-NAT and Per1, whereas its effect on Per2 was negligible. Contrary to what was observed in other systems, NE stimulation did not induce circadian expression of Per1. The effect of NE on Per1 level was dose- and receptor subtype-dependent, and both cAMP and cGMP induced Per1. Per1 was not induced by repeated NE - or forskolin - stimulation. Protein synthesis was not necessary for NE-induced Per1, but it was for reduction of Per1 following NE stimulation. Per1 transcription in pinealocytes was activated by BMAL1/CLOCK. Our results indicate that important differences are present in the regulation of these genes in the mammalian pineal. PMID- 12077486 TI - Hyperosmolality induced by betaine or urea stimulates endothelin synthesis by differential activation of ERK and p38 MAP kinase in MDCK cells. AB - We have previously shown that cultured porcine inner medullary collecting duct cells produce endothelin (ET) which suppressed arginine vasopressin (AVP)-induced cyclic adenosine monophosphate (cAMP) generation in an autocrine/paracrine feedback-like fashion. Moreover, hyperosmolality, e.g. induced by sodium chloride and urea, stimulated ET synthesis. Since others showed that hyperosmolality also activates mitogen-activated protein (MAP) kinases and that p38 MAP kinase facilitates cellular influx of betaine to protect the cell from high extracellular solute (urea) concentrations, we were tempted to investigate a potential interaction of MAP kinases with ET production in cultured MDCK cells in response to extracellular hyperosmolality induced by betaine and urea, respectively. Increased extracellular tonicity (602 +/- 8 vs. control of 323 +/- 3 mosmol/kg H(2)O) induced by betaine stimulated ERK and, more strongly, p38 kinase activity at 0.5-2 h of incubation with a rise in ET-1 synthesis to 1,713 +/- 68 vs. 378 +/- 51 fmol/mg protein/24 h under control conditions (p < 0.01). The p38 MAP kinase inhibitor SB203580 suppressed the rise in betaine-induced ET-1 synthesis by 91% to 494 +/- 38 fmol/mg protein/24 h, whereas the MEK/ERK inhibitor U0126 suppressed it moderately by 34%. Hypertonicity induced by urea moderately stimulated ERK but not p38 MAP kinase activity at 0.5-2 h and at 24-48 h and resulted in a modest rise in ET-1 synthesis to 681 +/- 61 fmol/mg protein/24 h (p < 0.05) which was significantly suppressed by U0126 to 484 +/- 16 fmol/mg protein/24 h. These results suggest that a functional interaction between the MAP kinases ERK and p38 MAP kinase and ET-1 synthesis is involved in betaine's protection of MDCK cells in vitro which may represent an in vivo mechanism of protection from hyperosmotic stress induced by high extracellular solute concentrations. PMID- 12077485 TI - Co-expressions of different opioid receptor types differentially modulate their signaling via G(16). AB - Combinations of two different types of opioid receptors - delta-, kappa-, mu opioid receptors (DOR, KOR, and MOR) and opioid receptor-like receptor 1 (ORL(1)) - were co-expressed with the alpha subunit of G(16) in COS-7 cells, and the ability of various selective agonists to induce activation of phospholipase Cbeta was examined. Nociceptin/orphanin FQ-induced response was enhanced when ORL(1) was co-expressed with MOR or KOR but not DOR. The kappa-agonist U50,488H induced a modest inositol phosphate formation when KOR was expressed alone or with MOR, but the response was attenuated when co-expressing with either DOR or ORL(1). It is suggested that the co-expressions of two different opioid receptor types indeed modify their downstream signaling events. PMID- 12077487 TI - Renoprotective mechanisms of angiotensin II antagonism in experimental chronic renal failure. AB - AIMS: We investigated angiotensin II and nitric oxide-cGMP pathway in the development of hypertension and renal damage in chronic experimental nephritis. METHODS: Rats with autoimmune nephritis were treated for 12 weeks with AT1 receptor antagonist L-158,809 and/or ACE inhibitor captopril given in drinking water. Blood pressure, urinary albumin, and urinary excretion of cGMP were measured. Renal density of ACE, AT1 and AT2 receptors was determined by quantitative in vitro autoradiography. RESULTS: L-158,809, captopril, and their combination decreased blood pressure and normalised urinary albumin excretion rate in rats with nephritis. In L-158,809-treated rats, cGMP excretion was increased compared to the vehicle-treated nephritic group suggesting that the dysfunctional nitric oxide system may be activated by angiotensin antagonism. In nephritic rats, AT1 and AT2 receptor binding densities in renal medulla were decreased, cortical AT receptor expression remained unchanged. Following L 158,809 treatment, both AT1 and AT2 receptor binding was suppressed. CONCLUSION: Long-term blockade of AT1 receptors in chronic nephritis has beneficial effects both on albuminuria and blood pressure being as effective as ACE inhibition or their combination. The stimulatory effect of AT1 receptor antagonism on cGMP production was not mediated by AT2 receptor-dependent mechanisms suggesting that AT1 receptor blockade per se favours activation of humoral pathways that stimulate cGMP production and potentially contribute to renal protection in chronic nephritis. PMID- 12077488 TI - Results from the TIP (Tritace in Proteinuria) intensified monitoring project. AB - Albuminuria has been shown to identify patients with an increased cardiovascular risk, and in clinical studies ACE inhibitors reduce the urinary protein excretion. It was the primary aim of this intensified monitoring project to determine whether these results can be reproduced in a clinical practice setting. Micro- (2.7-22.6 mg albumin/mmol creatinine) or macroalbuminuria (>22.6 mg/mmol) was confirmed by a central laboratory in 598 out of 773 patients with hypertension who had albuminuria >50 mg/l on a Micral Test II performed by 147 general practitioners. Coronary heart disease (prevalence rates 15% in patients with normalbuminuria, 33% in patients with microalbuminuria, and 40% in patients with macroalbuminuria), heart failure (prevalence rates 19, 29, and 32%, respectively), left ventricular hypertrophy (prevalence rates 30, 42, and 38%, respectively), and peripheral vascular disease (prevalence rates 7, 15, and 20%, respectively) were significantly more common in patients with elevated urinary albumin excretion. 230 patients with microalbuminuria and 202 subjects with macroalbuminuria were treated with the angiotensin-converting enzyme inhibitor ramipril for 6 months. The treatment significantly lowered mean arterial blood pressure (from a median value of 120 mm Hg, quartiles 113-125 mm Hg, to 103 mm Hg, quartiles 100-109 mm Hg) as well as urinary albumin excretion (from a median value of 18 mg/mmol creatinine, quartiles 7.2-54.6 mg/mmol creatinine, to 6.5 mg/mmol creatinine, quartiles 1.6-23.1 mg/mmol creatinine). The treatment efficacy was unaffected by age, body mass index, and smoking status. Patients with diabetes mellitus type II and heart failure also had a significant, although less pronounced reduction of albuminuria. In summary, we conclude that ramipril is able to reduce the urinary albumin excretion in a clinical practice setting, as has been shown in clinical studies. However, the treatment response is not completely uniform, as special patient populations seem to be more resistant to therapy. PMID- 12077489 TI - Endothelial nitric oxide synthase affects the progression of autosomal dominant polycystic kidney disease. AB - BACKGROUND/AIM: The phenotypic variability of autosomal dominant polycystic kidney disease (ADPKD) cannot be explained only by various mutations of two known genes (PKD1 and PKD2), but the influence of other unknown factors should also be considered. Impairment of endothelial function has been observed in ADPKD patients. Our study tried to verify the hypothesis that endothelial nitric oxide synthase (ecNOS) gene polymorphisms in intron 4 could have some relevance to the progression of ADPKD. METHODS: We examined 155 Czech patients with ADPKD (80 males, 75 females; mean age 43.4 +/- 14.7 years) and 100 genetically unrelated healthy subjects (50 men, 50 women; mean age 51.2 +/- 8.2 years). The genomic DNA was amplified by polymerase chain reaction and the products were separated on 1.5% agarose gel and visualised by ultraviolet transillumination. We compared subjects homozygous for the ecNOSb allele with subjects homozygous and heterozygous for the ecNOSa allele. RESULTS: The frequencies of ecNOSb/b, ecNOSa/b and ecNOSa/a were 81% (81/100), 17% (17/100) and 2% (2/100), respectively, in the control group, 60% (30/50), 32% (16/50) and 8% (4/50), respectively, in ADPKD patients with end-stage renal failure (ESRF), and 68.5% (72/105), 28.6% (30/105) and 2.9% (3/105), respectively, in ADPKD patients with normal renal function. The two-tailed t test revealed that the frequency of the ecNOSa allele (ecNOSa/b and ecNOSa/a) in dialysed patients was significantly higher than in ADPKD patients with normal renal function (p < 0.05). CONCLUSION: The a allele of the ecNOS gene polymorphism showed a significantly higher incidence among patients with ESRF caused by ADPKD. The a allele might have a negative influence on the progression of ADPKD. PMID- 12077490 TI - Effect of endovascular treatment on nitric oxide and renal function in Takayasu's arteritis with renovascular hypertension. AB - BACKGROUND: Renal involvement in Takayasu's arteritis (TA) effects the disease outcome and endovascular treatment is an effective treatment of choice. We investigated nitric oxide (NO) levels and the effect of endovascular treatment in renovascular hypertensive TA patients. METHODS: In five hypertensive patients with renal artery stenosis due to TA, serum creatinine, nitrite, nitrate; urinary microalbumin, nitrite, nitrate measurements and blood pressures were recorded at entry and after 24 h and 6 weeks of endovascular treatment. RESULTS: Serum NO levels were higher in patients than controls (p = 0.008). Serum and urine NO levels increased 24 h after the treatment and decreased after 6 weeks (p = 0.015; p = 0.01, respectively). After the treatment blood pressures decreased. Urinary microalbumin excretions increased after the intervention (p = 0.02) and returned to normal in patients 1 and 4, and decreased in the others. There were no significant differences in estimated glomerular filtration rate (EGFR), serum creatinine, urinary sodium and potassium levels. CONCLUSION: Increased NO secretion in these patients may contribute to improve the prognosis of renal function through its vasodilator and antiproliferative activities possibly by counterbalancing the excessive vasoconstrictor actions. Endovascular treatment causes a dilatation-induced shear stress that may be responsible for the increased NO release, which in turn leads to the rapid hypotensive response. PMID- 12077491 TI - Comparison of dialysis and clinical characteristics of patients with frequent and occasional hemodialysis-associated hypotension. AB - BACKGROUND: Symptomatic dialysis hypotension (DH) continues to be a common problem. By comparing patients prone and resistant to DH, several dialysis session and patient related characteristics have been identified that confer susceptibility to DH. Less is known, however, about the comparison of patients with frequent and only occasional DH. The aim of the study was to compare clinical and dialysis-session- (complicated by hypotension) related data between those with frequent (fDH) and those with occasional dialysis hypotension (oDH). METHODS: Nine hundred and fifty-eight patients at 11 dialysis units were followed for 10 months and characteristics of patients with fDH (> or = 10 hypotensive events necessitating medical intervention) (n = 96) were compared to that of patients with oDH (1 or 2 events/10 months) (n = 130). Significant and independent predictors of fDH were obtained by multivariate logistic regression. RESULTS: Significant differences between fDH vs. oDH patients were older age (64.4 vs. 56.9 years, p < 0.001), more females (66 vs. 46%, p < 0.005) in fDH. More fDH patients had diabetes (27 vs. 15%, p < 0.05) and less had glomerulonephritis (15 vs. 35%, p < 0.001) as the cause for ESRD. Coronary artery disease (68 vs. 50%, p < 0.01) and long-acting nitrate treatment (51 vs. 30%, p < 0.001) was more frequent while treatment with ACEI (33 vs. 48%, p < 0.05) or Ca channel blockers (40 vs. 53%, p < 0.05) were less frequent in patients with fDH. Patients with fDH had higher serum phosphorus levels (1.99 vs. 1.79 mmol, p < 0.005). Dialysis session related data were similar but the hypotensive episode occurred earlier during dialysis in fDH (136 vs. 156 min, p < 0.01). In multivariate analysis, significant independent predictors of fDH were older age (OR = 1.04 [1.02-1.07]), lack of glomerulonephritis as renal diagnosis (2.63 [1.18-5.87]), high phosphorus levels (5.0 [2.45-10.0]), lack of use of Ca-channel blockers (2.09 [1.12-3.91]), and the use of nitrates (2.38 [1.24-4.55]). CONCLUSION: Features of the dialysis sessions complicated by DH seem to be similar between patients with fDH and oDH, while patient characteristics such as older age, renal diagnosis other than glomerulonephritis, higher serum phosphorus levels, use of nitrates, and lack of use of calcium channel blockers are significantly and independently associated with fDH. PMID- 12077492 TI - Mean platelet volume and coronary heart disease in hemodialysis patients. AB - BACKGROUND: Mean platelet volume (MPV) has been shown to be an independent risk factor for myocardial infarction in the general population. METHODS: In the present cross-sectional study it was examined whether MPV may be associated with coronary heart disease (CHD) in patients in chronic hemodialysis treatment (n = 518). CHD was diagnosed if there was a history of myocardial infarction, coronary artery bypass grafting, or critical stenosis on coronary angiography. RESULTS: Mean platelet volume in hemodialysis patients was 8.22 +/- 0.06 fl. As compared with hemodialysis patients with a MPV <7.168 fl (the lower quintile limit), significantly more hemodialysis patients with a MPV >9.172 fl (the upper quintile limit) had CHD (odds ratio, 1.29; 95% confidence interval, 1.02-1.64). In a stepwise multiple logistic regression analysis MPV appeared to be an independent factor of CHD together with age, smoking, predialysis systolic blood pressure and serum triglyceride concentration. In a stepwise multiple linear regression model, MPV significantly depended on platelet count, volume of ultrafiltrate removed in one hemodialysis session, white blood cell count, treatment with beta blockers, heart rate, serum sodium and serum potassium concentrations. Additional measurements in 30 patients showed that MPV after a single hemodialysis session was not significantly different compared to predialysis values. CONCLUSION: From the present study it is concluded that MPV may be associated with CHD in hemodialysis patients. PMID- 12077493 TI - Outcome of dialysis access-related septicemia among diabetics following optimized AV-fistula placement. AB - BACKGROUND: AV fistula (AVF) is the safest of vascular accesses with lowest infection rates; yet only 23% patients used AVF during 1997 in USA. The lower prevalence of AVF among diabetics on hemodialysis (HD) places them at a higher risk of vascular-access-related septicemia (VRS) and ensuing mortality. In this study we assessed the outcome of VRS after maximizing the frequency of native AVF in this largest growing population on HD. METHODS: Study included 218 patients, 63 diabetics and 155 nondiabetics on HD, through July 1996 to July 2000 when National Kidney Foundation-Dialysis Outcome and Quality Initiative (NKF-DOQI) set goal was accomplished with overall 72% of functioning AVF (57.2% diabetics and 78.1% nondiabetics) through joint efforts of nephrologists and vascular surgeons. RESULTS: Overall, 10.6% patients per year developed VRS through 125 episodes, over 10,464 patient-months, recording 1.19 episodes per 100 patient-months. In the diabetic group, 13.87% patients per year had VRS during 44 episodes with 1.45 episodes per 100 patient-months while 1.08 episodes per 100 patient-months were recorded in nondiabetics with 9.35% per year having VRS during 81 episodes. Collectively, catheters recorded 1.5 folds higher VRS episodes in diabetic than in nondiabetic group. Mortality of 9.28% per year in diabetic group as compared to that of 6.45% per year in nondiabetic group [RR-1.436, 95% CI (0.778-2.651)] was observed, while overall mortality of 7.5% per year recorded is a good deal lower than 12-22% reported. CONCLUSION: The NKF-DOQI set aim of dialyzing over 50% patients through AVF is attainable in diabetics as well. Optimizing AVF is a viable approach to lessen VRS related mortality in diabetics on HD. Our continued dependence on vascular catheters is largely responsible for higher mortality in diabetics than nondiabetics on HD due to lack of cagily established pre-ESRD program for diabetics. PMID- 12077495 TI - Serum insulin-like growth factor 1 level and nutritional assessment in nondialytic patients with chronic renal failure. AB - AIM: In order to understand the nutritional status of nondialytic patients with chronic renal failure (CRF), nutritional assessment was made in 20 nondialytic patients (15 males and 5 females; mean age 43.7 +/- 15.1 years). METHODS: Twenty CRF inpatients were selected for nutritional assessment, and 20 normal subjects served as controls. The serum insulin-like growth factor 1 (IGF-1) concentration was measured by ELISA. Serum albumin, prealbumin, and transferrin levels were also determined. RESULTS: The mean IGF-I and transferrin levels in the CRF patients were significantly lower than those in normal subjects (IGF-1: 176.2 +/- 92.5 microg/l vs. 266.7 +/- 101.7 microg/l, p < 0.01; transferrin: 2.57 +/- 0.58 g/l vs. 3.18 +/- 0.27 g/l, p < 0.05). The IGF-1 levels in 7 patients with a serum albumin concentration <40.0 g/l were significantly lower than those in 13 patients with a serum albumin concentration >40.0 g/l (95.6 +/- 42.4 microg/l vs. 219.6 +/- 82.7 microg/l, p < 0.01). The IGF-1 levels in cases treated with alpha ketoacid were higher than in those without alpha-ketoacid treatment. The IGF-1 levels were positively correlated with creatinine clearance (r = 0.7066, p < 0.01) and serum transferrin concentration (r = 0.5347, p < 0.05). CONCLUSIONS: The fact that serum IGF-1 was correlated with serum transferrin and creatinine clearance suggests that IGF-1 may be a good indicator for assessing the nutritional status of CRF patients. The serum IGF-1 level in CRF patients is probably lower than that in normal subjects and could be improved by nutritional therapy. PMID- 12077496 TI - Role of complement in tubulointerstitial injury from proteinuria. AB - The complement system is being increasingly implicated in the pathogenesis of progressive renal disease resulting from persistent proteinuria. It has recently been established that renal tubular cells can produce complement, activate complement, and respond to complement activation. Complement proteins that pass through the glomerular barrier along with other serum proteins in the proteinuric state may become activated at the tubular epithelial brush border, and lead to a cascade of events culminating in cell injury. Furthermore, nephrotic components in the urinary space may cause direct activation of the tubular cells to overexpress complement and contribute to local tissue injury. In this review, we will discuss the current evidence supporting a role for complement in the pathogenesis of progressive tubulointerstitial damage in the proteinuric state. The possibility of complement inhibition intervening in progressive tubulointerstitial injury due to proteinuric glomerular disease will also be considered. PMID- 12077497 TI - [Prenatal medicine--claims and reality]. PMID- 12077498 TI - [What is the risk? Perspectives of prenatal diagnosis]. AB - The term 'risk' may be seen under multiple aspects. First of all, there is the risk of giving birth to a disabled child. Secondly, prenatal diagnosis itself constitutes a risk. Last but not least, there is a risk for society when presuming that everything is feasible. However 'risk' does not only mean danger, it also implies the notion of venture and responsibility. Responsibility towards society consists in opposing wrong developments. Responsibility towards the disabled child is often a pretext for not being able to bear misery. Life has to be lived; each other concept is contrary to nature. The only justification for not running the risk of having a disabled child lies in the responsibility towards one's own family. But often it is only in view of the task that we are assigned that we come to know our fortitude. PMID- 12077499 TI - [Aporia of prenatal medicine]. AB - Prenatal medicine touches upon elementary questions of the quality of life - of the mother as well as of the unborn child and of the physician. In essence, it is always about the determination of human worth by human beings. Each prenatal care examination is a measure of prenatal medicine. Prenatal medicine has gained today, and rightly so, a high and positive position in obstetrics and within our society. Without a doubt however, the advancements of prenatal medicine have also awakened demands and desires. We are increasingly faced with the 'demand for a healthy child'. In the ethical evaluation, intrauterine diagnostic measures are questionable as long as no intrauterine therapy is possible and only the killing of the child follows as 'therapy'. The contents of the 'indication', especially with respect to what can reasonably be demanded from the mother or the parents, are discussed with respect to liability law - 'the child as damage'. The reform of 218 StGB (German criminal law code) has provoked, among other issues, the pre- and postnatal problem of iatrogenic preterm birth. The ever present question is becoming more transparent with the liveborn sick child, which should have actually died, i.e. why instead the life of this child suddenly assumes the full legal protection so that active handling is replaced by passiveness. The two facedness of prenatal medicine appears even more clearly when picturing the current political discussions concerning what is newly feasible in prenatal medicine, namely preimplantation diagnostics. PMID- 12077500 TI - [Obstetrical ultrasonography]. AB - BACKGROUND: As with all new diagnostic options in medicine, great hope was placed in the introduction of high-resolution prenatal sonography. Progress tends to induce higher expectations. Patients and doctors alike may initially overestimate the possibilities of medical advances. QUESTION: The question at hand is whether we can validate a positive influence of prenatal ultrasound on the fetal and/or maternal pregnancy outcome, and how a sonographic routine screening should be structured. MATERIAL AND METHODS: This is an attempt to survey the heterogeneous pool of internationally published data with regard to these critical questions. RESULTS AND CONCLUSION: Different studies lead to a positive conclusion on the benefits of ultrasound monitoring in pregnant women. Ultrasound screening has a positive effect on medical as well as economic issues. Nevertheless litigation concerning prenatal diagnosis remains a growing problem. PMID- 12077501 TI - [Non-invasive prenatal diagnosis--already a reality?]. AB - The genetic analysis of fetal cells enriched from the blood of pregnant women has been a long-standing goal of those researchers and clinicians who want to use this technology for the development of a non-invasive risk-free method for prenatal diagnosis. Recent large-scale studies, such as the NIH 'NIFTY Study', have reiterated that promise of this approach, but have reinforced also the consensus amongst researchers that current technology is not yet sufficiently advanced to permit routine use. The new observation of cell-free fetal DNA in maternal plasma has been eagerly seized upon by several research groups, who have shown in independent large-scale studies that this technology can be used for the extremely reliable identification of facile fetal genetic traits such as the fetal RhD status. Consequently, this approach has been the first to make the transition from the laboratory 'bench' to the 'bed'. PMID- 12077503 TI - The influence of family history of stuttering on the onset of stuttering in young children. AB - Content analysis was undertaken of the case records of 61 children who stuttered who were attending a specialist centre for children who stutter. The subjects were divided into two groups, on the basis of family history of stuttering. Positive family histories of stuttering were reported for 44 children and 17 had negative family histories. The two groups were compared in terms of gender ratios, the age of onset and the type of onset (gradual vs. sudden) of stuttering. Those with positive family histories began stuttering earlier than those with no reported family history of stuttering, though this difference was not statistically significant. The type of onset of stuttering was not related to the presence or absence of a family history of stuttering. The findings are discussed in terms of the practicability of this method of data collection. PMID- 12077504 TI - Acoustic characteristics [correction of characeristics] of adolescent actors' and non-actors' voices. AB - The aim of the present study was to investigate differences in the acoustic characteristics of the voice between adolescent actors and non-actors. The experimental sample consisted of 10 actresses and 10 actors while the control sample included 13 girls and 14 boys. Phonation of the vowel /a/, spontaneous speaking and oral reading provided a set of acoustic variables (fundamental frequency, jitter, shimmer, speaking and reading ranges); t test showed statistically significant differences between actresses and non-actresses in speaking range, reading F(0) maximum, and reading range, whereas between actors and non-actors the difference was found in reading range only. The results showed that drama in education without systematic voice training had no effects in terms of acoustic characteristics of the voice. PMID- 12077505 TI - Differences in glottal opening pattern between Tokyo and Osaka dialect speakers: factors contributing to vowel devoicing. AB - In Japanese, high vowels /i/ and /u/ are often devoiced when they are preceded and followed by voiceless consonants. The phenomenon is called vowel devoicing. The frequency of its occurrence is high in dialects of eastern Japan including standard (Tokyo) Japanese and low in dialects of western Japan including Osaka dialect. It has been claimed that consonants, but not vowels, are pronounced more carefully in eastern dialects compared to western dialects, which results in higher frequency of vowel devoicing. This paper examines whether the glottal opening gesture of voiceless consonants differs between those Tokyo speakers who consistently show vowel devoicing and Osaka speakers who do not show vowel devoicing. The results indicated that the duration of glottal opening for /k/ in voiced CV syllables was significantly longer for Tokyo speakers than for Osaka speakers. For Tokyo speakers, duration and maximum size of the glottal opening in devoiced /kite/ are significantly larger than /k/ in voiced syllables. This difference cannot be explained by the simple addition of the glottal gestures of [k] and [t]. Furthermore, the difference in vowel length between /i/ and /e/ was related, at least in part, to the difference in the amount of the airflow passing through the vocal tract about the time the stop closure was released. PMID- 12077506 TI - Dynamic extremes of voice in the light of time domain parameters extracted from the amplitude features of glottal flow and its derivative. AB - In the first part of the study, the subjects (4 females, 4 males) produced sustained breathy, normal and pressed types of phonation. The maximum AC flow (f(ac)) and the negative peak amplitude (d(min)) of the first derivative of the glottal flow were extracted from glottal volume velocity waveforms estimated by inverse filtering. The ratio between f(ac) and d(min) (amplitude quotient, AQ), which gives an estimate of the closing time of the glottal flow, and its normalised version (NAQ) were computed. The NAQ values correlated strongly with the closing quotient of glottal flow. In the second part of the study, the aim was to investigate the dynamic extremes of the human voice in terms of F(0), SPL, f(ac), d(min), AQ and NAQ as well as the intraoral pressure (p) used for subglottal pressure estimation. Eleven normal subjects (5 females, 6 males) participated in this experiment. The subjects started with the production of soft phonation (approximately 55 dB at 40 cm) and increased their vocal intensity at 5 dB steps up to the level of 105 dB. At the analysis stage, the voice samples were divided into three categories: soft (<70 dB), normal (70-90 dB) and loud (>90 dB). In soft and normal phonation, intensity change was correlated with changes in the shape of the glottal flow waveform, whereas the loud voice samples showed a large increase in F(0) and p values. At the loudest extreme, d(min) approached its maximum and AQ its minimum values and NAQ started to rise. In terms of the NAQ values of stressed syllables, the type of phonation was breathy in the soft intensity category, whereas in the normal and loud intensity categories it was within the range of the pressed type of phonation of sustained phonation samples. PMID- 12077507 TI - Effect of cleft type on overall speech intelligibility and resonance. AB - The main purpose of this study was to measure and compare the effect of an unilateral cleft lip and palate (UCLP) and a bilateral cleft lip and palate (BCLP) on overall speech intelligibility, nasalance and nasality. An additional objective was to compare the nasalance and nasality patterns of cleft palate children with the available normative data for normal children. The subjects were 37 children with an UCLP (19/37) or a BCLP (18/37) with normal cognitive and motor functions and normal hearing levels. All subjects had a non-syndromic cleft, followed the same surgical protocol and did not undergo secondary pharyngeal surgery. To measure and compare the effect of cleft type, objective and subjective assessment techniques were used. For the objective assessment of nasal resonance the nasometer and the mirror-fogging test were used. Nasalance scores were obtained, while patients produced sounds, read three standard passages or repeated sentences focusing on specific consonants. A perceptual evaluation of speech, the Gutzmann (1913) test and the tests of Bzoch (1989) were used as subjective assessment techniques. The perceptual assessment of the overall speech intelligibility was based on the spontaneous speech and reading or repeating of the nasometric sentences. These samples were judged by three speech pathologists using a 4-category nominal scale. A subjective test (Morley, 1945) was used to evaluate the velopharyngeal mechanism. The findings of the present study show no statistically significant differences for nasalance values, nasality and overall speech intelligibility between the UCLP and BCLP children. Significant differences were found between the data of the normal group and the cleft palate children. These differences included nasalance values as well as nasality data and overall speech intelligibility. PMID- 12077508 TI - Comparison of regional blood flow values measured by radioactive and fluorescent microspheres. AB - Fluorescent microspheres (FM) have become an attractive alternative to radioactive microspheres (RM) for the measurement of regional blood flow (RBF). The aim of the present study was to investigate the comparability of both methods by measuring RBF with FM and RM. Eight anaesthetised pigs received simultaneous, left atrial injections of FM and RM with a diameter of 15 microm at six different time points. Blood reference samples were collected from the descending aorta. RBF was determined in tissue samples of the myocardium, spleen and kidneys of all 8 animals. After radioactivity of the tissue samples was determined, the samples were processed automatically for measuring fluorescence using a recently developed filter device (SPU). RBF was calculated with both the isotope and spectrometric data of both methods for each sample resulting in a total of 10,512 blood flow values. The comparison of the RBF values yielded high linear correlation (mean r(2) = 0.95 +/- 0.03 to 0.97 +/- 0.02) and excellent agreement (bias 5.4-6.7%, precision 9.9- 16.5%) of both methods. Our results indicate the validity of MS and of the automated tissue processing technique by means of the SPU. PMID- 12077509 TI - The influence of node-fibril morphology on healing of high-porosity expanded polytetrafluoroethylene grafts. AB - PURPOSE: To manufacture high-porosity expanded polytetrafluoroethylene (ePTFE) vascular grafts with the same internodal distance but different node-fibril morphology, and to evaluate their biologic behaviors in a canine carotid artery implantation model. MATERIALS AND METHODS: Several types of high-porosity ePTFE vascular grafts with the same inside diameter (4 mm) and wall thickness (650 microm) were manufactured under different heating, stretching conditions. The luminal surface and cross section of the grafts were photographed by scanning electron microscopy and the node-fibril structure was examined. Two typical types of high-porosity ePTFE vascular grafts were then selected and proceeded to an animal study. The test grafts were explanted after an interval of 12 weeks and subjected to histomorphometric analyses. RESULTS: The following two types of high porosity ePTFE vascular grafts were selected; one had a through-pore structure extending from the outer to the inner surface and the other had a random-node architecture with tortuous path channels extending from the outer to the inner surface. The histomorphometric analyses of thrombus-free surface, thickness of pseudointima, cellular ingrowth, capillary ingrowth, and cellular proliferation revealed no significant differences between the grafts. CONCLUSION: In high porosity ePTFE vascular grafts, graft healing enhanced by transmural tissue ingrowth may be not largely dependent on node-fibril morphology. This knowledge will be helpful to design a new type of high-porosity ePTFE vascular grafts available for clinical use. PMID- 12077510 TI - Connective tissue growth factor in indomethacin-induced rat gastric ulcer. AB - The healing of gastric ulcers requires not only the complete epithelial covering but also the restitution of connective tissue. Transforming growth factor-beta (TGF-beta) and its downstream mediator, connective tissue growth factor (CTGF), are potent stimulators for connective tissue formation during wound healing. The expression of TGF-beta, CTGF and type III collagen mRNA in indomethacin-induced gastric ulcers in rat, was investigated by Northern blot analysis. We also examined the localization of CTGF producing cells by in situ hybridization. Northern blot analysis showed expression of TGF-beta mRNA on days 1 and 3 after indomethacin administration, expression of CTGF mRNA on days 1, 3 and 7 and type III collagen mRNA expression on days 1, 3, 7 and 12, respectively. Control animals showed no expression of TGF-beta, CTGF or type III collagen mRNA. In situ hybridization showed CTGF mRNA positive cells on days 1, 3 and 7 after ulcer induction in fibroblast-like cells and in some of the blood vessels. Thus our findings indicate that growth factor CTGF, together with TGF-beta, participates in gastric ulcer healing by regulating connective tissue formation and angiogenesis. These results are compatible with the role of CTGF as a downstream mediator of TGF-beta effects. PMID- 12077511 TI - Prevention of intraabdominal adhesions produced by polypropylene mesh. AB - Polypropylene mesh (PPM) is widely used in ventral hernia repair, however is also associated with visceral adhesions when the mesh is exposed to intraabdominal organs. In this study, a composite mesh [ePTFE (expanded polytetrafluoroehylene) + PPM] and sodium hyaluronate/carboxymethyl cellulose (NaHA/CMC) membrane laid under PPM are evaluated in terms of adhesion formation in a rat model of ventral hernia repair. In this experimental study, a 2 x 1 cm of peritoneum and underlying muscle defect was created at the right side of the anterior abdominal wall of 37 male Wistar albino rats. These defects were repaired with 2.5 x 2 cm PPM in group 1 (n = 13), with composite mesh in group 2 (n = 12) and with PPM+NaHA/CMC in group 3 (n = 12). Rats were sacrificed after 14 days and the prosthetic materials were examined for the calculation of surface area percentage covered by adhesions, for organ involvement in the adhesions and for histological evaluation. There was a reduction in the adhesion-covered area in group 2 and group 3. Organ involvement was predominantly seen in group 1. Neoperitoneum was perfect in group 2. Fibrosis and inflammation were higher in group 1. All groups showed adhesiogenic potential to some extent. This potential was maximum with PPM. Both ePTFE and NaHA/CMC displayed similar effects in preventing adhesion formation. PMID- 12077512 TI - Na(+)/H(+) exchanger inhibitor HOE642 offers myoprotection in senescent myocardium independent of ischemic preconditioning mechanisms. AB - BACKGROUND: Inhibition of Na(+)/H(+) exchange has been shown to provide functional protection during ischemia and reperfusion in mature heart. This study was undertaken to elucidate the effect of Na(+)/H(+) exchange inhibitor HOE642 in the aged rabbit heart. METHODS: Isolated rabbit hearts were subjected to 1 h of left descending coronary artery (LAD) ischemia and 1 h of reperfusion. To determine the effects of HOE642 on ischemia/reperfusion injury, seven aged or mature hearts received the Na(+)/H(+) exchange inhibitor HOE642 (1 microM) for 15 min before the ischemia and for 30 min after reperfusion. Seven aged (more than 135 weeks) or mature (15-20 weeks) rabbit hearts served as a control (untreated) with no interventions. Left ventricular pressures, monophasic action potentials and coronary flows were measured throughout the experiment and infarct size was detected at the end of experiment. RESULTS: (1) In the mature hearts, HOE642 improved postischemic functional recovery (63.1 +/- 5.0% vs. 84.4 +/- 5.4%, mature untreated vs. mature HOE, p < 0.05) and reduced infarct size as compared to untreated hearts (42.0 +/- 2.5% vs. 24.8 +/- 2.3%, mature untreated vs. mature HOE, p < 0.05). (2) Although infarct size in aged untreated hearts was significantly decreased as compared to mature untreated hearts (42.0 +/- 2.5% vs. 19.3 +/- 1.6%, mature untreated vs. aged untreated, p < 0.05), there are no significant differences regarding postischemic functional recovery between mature and aged untreated hearts (63.1 +/- 5.0% vs. 59.5 +/- 5.9%, mature untreated vs. aged untreated, p = n.s.). (3) In the aged hearts, HOE642 improved postischemic functional recovery as compared to untreated hearts (59.5 +/- 5.9% vs. 85.9 +/- 8.1%, aged untreated vs. aged HOE, p < 0.05). CONCLUSION: Na(+)/H(+) exchange inhibitor HOE642 is effective against ischemia-reperfusion injury in senescent as well as mature hearts. PMID- 12077513 TI - Kupffer cell targeting by intraportal injection of the HVJ cationic liposome. AB - The aim of this study was to target Kupffer cells (KCs) selectively and efficiently by the intraportal injection of fusigenic cationic liposomes with hemagglutinating virus of Japan components (HVJ cationic liposomes). Phosphorothioate FITC-oligodeoxynucleotides (FITC-ODNs) encapsulated in either HVJ cationic liposomes, HVJ anionic liposomes or conventional cationic liposomes without HVJ were transferred to the rat. FITC-ODNs in HVJ cationic liposomes administered via portal vein were selectively transfected to KCs for up to 24 h with no apparent cytotoxicity at higher transfection efficiency than FITC-ODNs in conventional cationic liposomes without HVJ administered via portal vein or tail vein. On the other hand, FITC-ODNs in HVJ anionic liposomes were observed mainly in hepatocytes, not KCs. This new method will be useful for the modulation of KCs activity in both basic research and clinical applications. PMID- 12077514 TI - Prolonged suppression of humoral immune response after organ transplantation. AB - Rats receiving either kidney or liver grafts were vaccinated with recombinant hepatitis B virus (HBV) vaccine either 1 or 10 weeks post-transplantation to explore the nonspecific immunosuppressive effect of transplantation surgery on antibody development. Response rate and geometric mean titer of the anti-HBs antibody were significantly lower in organ-transplanted animals undergoing early vaccination compared to unoperated control animals. Both parameters were affected when vaccination was performed early after kidney transplantation, but also in case of late vaccination in liver graft recipients. The nonspecific suppressive effect of transplantation surgery on the humoral immune response was related to the time point of vaccination after surgery and the extent of the surgical intervention and lasted longer than previously reported. PMID- 12077515 TI - Reimplanted ureter as an alternative to the catheterizable Mitrofanoff tube. AB - OBJECTIVE: We performed this study in dogs to investigate whether a ureter can be used as an alternative to the appendix without disrupting the uniformity of the gastrointestinal system. MATERIALS AND METHODS: This study comprised 10 adult healthy female mongrel dogs. The procedure was done in 2 stages: in the first stage, we performed a left-end cutaneous ureterostomy at the lower left quadrant just above the inguinal ligament. A second operation was performed 6 weeks later. The left ureter was divided and using Paquin's technique, the proximal end of the distal third of the left ureter was reimplanted in the anterolateral surface near the dome of the bladder. A high transureteroureterostomy with the remaining left proximal ureter to the opposite ureter completed the reconstruction. The bladder neck was ligated in order to create an experimental bladder outlet obstruction. At the time of creating the distal ureteral stoma, a feeding tube was left in the bladder, passing through the ureterostomy stoma and sutured to the skin. This catheter remained in place for 20 days, preventing the bladder form distending. Then in the following 6 months, intermittent catheterization was applied through the stoma at 3- to 4-hour intervals. Exploration was performed at the end of the 6th month. RESULTS: No animal died. The dogs were still continent. Intravenous urography, urea and creatinin values were normal. The ureteral stoma was viable in all 10 cases. Stenosis of the ureteral stoma developed in 1 dog and required surgical revision. Possible traumatic effects were investigated by histological sections, but no harmful effects were found at the ureteral endothelium. CONCLUSION: This method may be an alternative to the Mitrofanoff method, which uses the appendix. PMID- 12077516 TI - Telemetric small intestinal motility recording in awake rats: a novel approach. AB - INTRODUCTION: Ideally, gastrointestinal motility recording should be done in freely moving, stress-free animals. However, no such method is currently available for rats. METHODS: Two NiCr electrodes were sutured to the jejunum and connected to an implantable electromyographic (EMG) transmitter in rats. EMG signals were radio-transmitted to a receiver placed at the bottom of the rats' home cages. RESULTS: Fasting and postprandial jejunal EMG signals could be detected by telemetry. Phase III contractions of the MMC were easy to identify visually and occurred at a rate of about 4.8 per hour. Feeding disrupted the phasic contraction pattern 15 min after the start of food intake and lasted for 2 h. The motility index (MI, area under the curve) was calculated and increased postprandially. CONCLUSION: Telemetric transmission of rat gastrointestinal EMG signals is feasible and results are comparable to those given in the literature. PMID- 12077517 TI - Is subungual melanoma related to trauma? AB - Subungual melanomas represent approximately 20% in dark-skinned and oriental populations compared to about 2% of cutaneous melanomas in white populations. UV exposure seems to be an important risk factor for cutaneous melanoma. However, UV radiation is unlikely to penetrate the nail plate. Another pathogenetic factor of subungual melanoma will be discussed. 406 subungual melanomas of the hands (n = 240) and feet (n = 166) of 74 patients from the melanoma registry of the Department of Dermatology, University of Tubingen, and of 332 patients from the literature were evaluated. The hypothesis of a uniform distribution of the occurrence of subungual melanoma on the fingers and toes had to be rejected (p < 0.001 using the chi(2)(4,0.95) test). There was a considerable predominance of subungual melanoma localized on the thumb (58% of all fingers) and the hallux (86% of all toes). Many patients report direct trauma related to the onset of subungual melanoma. This might be explained by coincidence, increased attention to a dark area under the nail, traumatic bleeding of a subclinical subungual melanoma or mutation of melanocytes during trauma-induced proliferation. Squamous cell carcinoma is known to occur in sites of chronic trauma. Trauma could be an etiologic factor in subungual melanoma as well. PMID- 12077518 TI - Genotype association of C(-735)T polymorphism in matrix metalloproteinase 2 gene with G(8002)A endothelin 1 gene with plaque psoriasis. AB - BACKGROUND: Excessive angiogenesis is one of the characteristic features of psoriasis. OBJECTIVE: To determine the possible genetic background of neo angiogenesis in plaque psoriasis, frequent polymorphisms in matrix metalloproteinase 2 (MMP-2) and endothelin 1 (ET-1) genes were studied. METHODS: The case group (n = 119) included patients with plaque psoriasis, aged 44 +/- 15 years. The age of onset of psoriasis was 27 +/- 11 years. The control group (n = 184) consisted of healthy subjects without any individual history of psoriasis, aged 37 +/- 15 years. C(-735)T MMP-2 and G(8002)A ET-1 polymorphisms were determined by PCR reaction with subsequent restriction analyses. RESULTS: A significant difference in genotype distribution of C(-735)T MMP-2 between psoriatic and control patients was found (p(corr) = 0.008). Two associated genotypes (CCGG and CTGG) of the two polymorphisms were significantly less frequent in psoriatic patients (p(corr) = 0.03 and p(corr) = 0.008, respectively). CONCLUSION: The results seem to reflect a different susceptibility of MMP-2 as well as of some associated MMP-2 and ET-1 genotypes to psoriasis. PMID- 12077519 TI - The spectrum of contact allergy in elderly patients with and without lower leg dermatitis. AB - BACKGROUND: There is a strong influence of lower leg dermatitis on the spectrum of allergic contact sensitivity in elderly patients. OBJECTIVE: To quantitatively assess the impact of this factor which confounds the age-related pattern of allergic contact sensitivity. METHODS: Stratified analysis of allergic contact sensitivity data collected between 1996 and 1999 in the 32 centres of the Information Network of Departments of Dermatology (three age groups: < or =60, 61 75, > or =76 years), both as crude and as directly standardised (for sex and leg dermatitis) proportions. RESULTS: Among the 36,070 patients tested with the German standard series, allergic contact sensitivity to neomycin, lanolin, cetostearylic alcohols, colophony, fragrance mix and balsam of Peru, among others, was (much) more common in the elderly, with proportions more or less affected by standardisation. CONCLUSION: The comparison of crude and standardised proportions provides evidence that allergic contact sensitivity to several allergens strongly depends not only on age itself, but on age-related and thus confounding co-morbidity. PMID- 12077520 TI - Exanthems in hospitalized pediatric patients: concordance between pediatric and dermatological diagnoses. AB - BACKGROUND: Exanthems in children often represent a diagnostic challenge. PURPOSE: To determine the concordance between pediatric and dermatological diagnoses of exanthems. DESIGN: Prospective study. PROCEDURE: Exanthems that appeared in hospitalized pediatric patients were diagnosed by pediatricians. Pediatric dermatologists, by consensus, either confirmed or modified the diagnoses. Whenever possible, laboratory tests were used to confirm the final clinical diagnoses. Age and evolution were compared with the Kruskal-Wallis test; the kappa coefficient was used to determine concordance. SETTING: Institutional tertiary referral pediatric care center. RESULTS: Concordance between pediatric and dermatological diagnoses was found in only 44 patients. When pediatric and dermatological diagnoses of exanthems were classified, the concordance between both diagnoses was very low (kappa = 0.165). CONCLUSIONS: Pediatric specialists, other than dermatologists, failed to diagnose common skin diseases. This may be a consequence of insufficient training in dermatology by medical schools and pediatric residency. PMID- 12077521 TI - Effects of Roxithromycin on the production of lipase and neutrophil chemotactic factor by Propionibacterium acnes. AB - BACKGROUND: The macrolide antibiotic roxithromycin is effective against acne associated with inflammation, but the mechanism by which this is achieved has not been clarified. OBJECTIVE: We studied the effects of roxithromycin on the production of lipase and neutrophil chemotactic factor by Propionibacterium acnes in vitro. RESULTS: Roxithromycin significantly inhibited the production of lipase and neutrophil chemotactic factor by P. acnes at a concentration one eighth of the MIC, at which the growth curve of P. acnes is not affected. CONCLUSION: One mechanism of the effectiveness of roxithromycin in acne therapy is thought to be the inhibition of bacterial lipase and neutrophil chemotactic factor production by P. acnes. PMID- 12077522 TI - Assessment of topical hypopigmenting agents on solar lentigines of Asian women. AB - BACKGROUND: So-called darkened age spots encompass distinct pathological processes. The efficacy of topical depigmenting agents is difficult to objectivate. OBJECTIVE: To assess the hypopigmenting effect of three cosmetic formulations using objective biometrological methods. METHODS: 50 women of South East Asian ancestry were enrolled in this pilot study. They had solar lentigines according to dermoscopic criteria. The lesions were treated by topical hypopigmenting formulations. Products were applied twice daily for 2 or 3 months. Assessments at 1-month intervals were made using narrow-band reflectance spectrophotometry, image analysis of video-recorded ultraviolet light reflection and photodensitometry- and image-analysis-assisted corneomelametry. RESULTS: A 20% azelaic acid formulation and another one containing 5% ascorbyl glucosamine, 1% kojic acid and alpha-hydroxyacid esters appeared inefficacious on solar lentigines. A stabilized soy extract showed a better although modest lightening effect when assessed by corneomelametry. The subclinical or faint mottled skin revealed by ultraviolet light examination better responded (p < 0.05) to treatments. CONCLUSION: Focal epidermal hyperpigmentation is better controlled by topical whitening agents when the increase in melanin content reflects a modest functional hyperactivity of melanocytes. PMID- 12077523 TI - Verrucae vulgares in children: successful simulated X-ray treatment (a suggestion based therapy). AB - BACKGROUND: Treatment of verrucae vulgares is sometimes difficult. Invasive methods should not be used for young children. OBJECTIVE: Evaluation of a special suggestion therapy for treatment of verrucae vulgares in children. METHODS: Nine children with warts on the hands and/or feet and in the face were treated with a simulated X-ray treatment. RESULTS: Five children showed a complete remission of warts, 3 children a partial remission. For 1 child, there was no response. On average, 3 treatment sessions were needed for children showing a complete remission. CONCLUSION: This therapy offers an easy-to-perform, alternative treatment option. It is noninvasive and does not depend on special psychological techniques for which training is required. PMID- 12077524 TI - Enhancement of classic Kaposi's sarcoma growth after intralesional injections of desferrioxamine. AB - We have previously shown that iron may be involved in the pathogenesis of Kaposi's sarcoma (KS) and that the iron chelator desferrioxamine (DFO) inhibits the growth and induces the apoptosis of KS cells in vitro. We treated an 85-year old man with classic KS with 5 weekly intralesional injections of DFO and observed the opposite effect in vivo. The DFO-treated lesion was characterised by the development of numerous KS papules within the drug diffusion area, whereas no change was noted in untreated or control saline-treated lesions. This suggests that intralesional iron chelators are not indicated in patients with KS. PMID- 12077525 TI - Multiple congenital symmetric skin dimples. AB - Skin dimples are seldom observed in sites other than the face. We report on a male premature child who was seen at the age of 2 months for the evaluation of cutaneous depressions symmetrically located on the shoulders, elbows and in the sacral region. Skin dimples have sometimes been considered a benign autosomal dominant trait. However, several authors have reported these cutaneous defects in a variety of conditions like congenital syndromes, infections, inborn errors of metabolism and mechanical trauma. In our case, the aetiology is unknown, even though maternal drug or infective exposure can reasonably be excluded as well as traumatic events. At a 3-year follow-up, the patient shows a normal psychophysical development. This appears to be the first case of a child presenting congenital, symmetric dimples in three different areas. PMID- 12077527 TI - Partial lipodystrophy in a patient with systemic lupus erythematosus. AB - A 54-year-old woman developed partial lipodystrophy on the left side of her face. She had been suffering from systemic lupus erythematosus (SLE) since 1985 when she was 45 years old, and she had been treated with 30 mg/day of oral prednisolone as an initial dose. Partial lipodystrophy appeared on her left lower jaw in 1994 when the SLE was inactive, and the dose of prednisolone was reduced to 5 mg/ 3 days. Gradually, the lipodystrophy spread toward her left cheek and her left forehead without any preceding skin symptoms. Histological examination showed a loss of fat tissue and mild lymphocytic infiltrations mainly around cutaneous appendages and vessels in the dermis and subcutaneous tissue. The dose of prednisolone was increased to 10 mg/day and the lesions stopped spreading. Such partial lipodystrophy is distinct from lipoatrophy of lupus profundus. PMID- 12077526 TI - Segmental neurofibromatosis. AB - Segmental neurofibromatosis is characterised by a limited, segmental distribution of cutaneous neurofibromatosis type 1 (NF1) lesions. It has been suggested that segmental NF results from a postzygotic NF1 gene mutation, and, recently, this hypothesis has been proven in a patient with regionally distributed cafe-au-lait (CAL) spots and freckles by demonstrating an NF1 microdeletion restricted to fibroblasts cultured from CAL spots. We describe here a patient with segmental NF in which we could not demonstrate any NF1 gene mutation in fibroblasts cultured from neurofibromas by use of the protein truncation test, enzymatic mutation detection and fluorescence in situ hybridisation. These data are in line with the concept that NF1 mutations in Schwann cells, but not in fibroblasts, correlate with neurofibroma formation. PMID- 12077528 TI - Levofloxacin is effective for inflammatory acne and achieves high levels in the lesions: an open study. PMID- 12077529 TI - How to remove ticks? PMID- 12077530 TI - Erythema multiforme due to rofecoxib. PMID- 12077531 TI - Cutaneous vasculitis related to celecoxib. PMID- 12077532 TI - The biobag - a new device for the application of medicinal maggots. PMID- 12077533 TI - Bullous eruption during chronic renal allograft rejection. PMID- 12077534 TI - Alopecia universalis associated with Zumbusch-type generalized pustular psoriasis. PMID- 12077535 TI - POEMS syndrome revealed by multiple glomeruloid angiomas. AB - POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, skin disorders) is a rare multisystemic disease associated with plasma cell dyscrasia. A 68-year-old woman with chronic renal insufficiency and arterial hypertension included in her medical history was admitted to the hospital with confusion, somnolence and asthenia. She presented ascites, hepatosplenomegaly, leg oedema, distal dysesthesias, leuconychia and multiple nodular purple red angiomas on the trunk, upper limbs and fingers. Hypothyroidism was revealed in the laboratory investigations and monoclonal IgG peak in immunoelectrophoresis. Electromyography showed both demyelinisating and axonal degenerative neuropathy. The diagnosis of POEMS syndrome was based on the dermatopathological examination of a cutaneous angioma; histology revealed features of glomeruloid angioma, a specific marker of this syndrome. PMID- 12077536 TI - Invasive tricholemmal carcinoma of the nose. AB - Tricholemmal carcinoma is a rare entity. It must be distinguished from the benign proliferating variant of the tricholemmal cyst. We report a case of tricholemmal carcinoma massively invading the nose. The neoplasm was excised surgically with a satisfactory outcome. PMID- 12077537 TI - Proteus syndrome of the hand. AB - Proteus syndrome exhibits an expanded phenotype. We report a case with restricted macrodactyly and cerebriform hypertrophy of one palm. The differential diagnosis with macrodactyly and other hamartomatous overgrowth syndromes is discussed. PMID- 12077538 TI - Tacalcitol ointment for long-term control of chronic plaque psoriasis in dermatological practice. AB - BACKGROUND: Various studies have shown the benefit of topical vitamin D(3) analogues in the treatment of mild to moderate plaque psoriasis. OBJECTIVE: Assessment of the efficacy, tolerability and safety of tacalcitol ointment in the long-term treatment of chronic plaque psoriasis in daily dermatological practice. DESIGN AND METHODS: In this open, multicentre, clinical phase IV study with a pre/post comparison design, 157 patients with chronic plaque psoriasis were included. Patients showing chronic plaque psoriasis, covering 7-20% of their body area, were treated with tacalcitol ointment (4 microg/g, Curatoderm) once daily and were assessed at baseline and monthly during the 6-month treatment period. The efficacy parameter psoriasis area and severity index (PASI) and total body surface involvement were assessed at each visit. Laboratory parameters were assessed at the beginning and at the end of the study. Adverse events were recorded at each visit. RESULTS: The mean PASI score decreased by 67%, and a marked reduction in sum scores of erythema, infiltration and desquamation was detected. The body area affected declined by 33% from 13.3 to 8.8%. There were no changes in laboratory parameters, and no case of hypercalcaemia was observed. No serious adverse events occurred during the study period. The recorded local side effects were usually transient and mainly mild. CONCLUSION: Tacalcitol ointment is safe, well tolerated and provides a further option for patients with psoriasis up to 20% body surface affected. Tacalcitol treatment can be recommended as effective therapy for long-term control of chronic plaque psoriasis in dermatological practice. PMID- 12077539 TI - Skin tensile properties in patients treated for acromegaly. AB - BACKGROUND: Somatotropic effects are described in the skin. Indeed, acromegaly is in part clinically recognized by cutaneous coarsening. The actual changes in tensile properties associated with the cutaneous manifestations are largely unknown. OBJECTIVES: To study the relationships between the skin tensile properties and the severity of acromegaly as assessed by serum levels of growth hormone (GH) and insulin-like growth factor 1 (IGF-1). PATIENTS AND METHOD: Assessments were made in 13 patients with acromegaly treated by somatostatin agonists combined or not with surgery. A total of 39 age- and sex-matched healthy subjects served as controls. Skin tensile properties were measured on the forearm and nape of the neck using a computerized suction device. RESULTS: Significant differences were yielded between the skin tensile properties in patients and normal subjects. The highest IGF-1 values in the patients' medical records were positively correlated with both skin distensibility and biologic elasticity. The most recent IGF-1 serum levels were negatively correlated with the visco-elastic ratio. No correlations were yielded between any of the biomechanical parameters and GH levels, disease duration and treatment dosages, respectively. CONCLUSION: The skin in acromegaly appears to be functionally more redundant and elastic than normal skin. The biomechanical changes appear quite different from those observed in other diseases with collagen deposition such as diabetes mellitus and scleroderma. PMID- 12077540 TI - Treatment of Kaposi's sarcoma with human chorionic gonadotropin. AB - Clinical-grade preparations of human chorionic gonadotropin (hCG) have been shown to be toxic to Kaposi's sarcoma (KS) cells. However, the mechanism of the anti-KS activity achieved with these preparations remains unclear. The results of clinical studies using commercial hCG preparations in human KS are also highly contradictory. The apparent controversy between different studies may be due to the fact that pro- and anti-KS components are present in varying proportions in different hCG preparations. As certain hCG preparations could not only lack the ability to control KS but also contain some contaminant KS growth factor(s), we suggest a cautious use of crude hCG for the treatment of KS. PMID- 12077541 TI - Allergic contact dermatitis from cosmetics. Retrospective analysis of 819 patch tested patients. AB - BACKGROUND: Cosmetics have been used since the oldest known civilisations, and nowadays almost everybody resorts to beauty products. OBJECTIVE: Considering the increasing incidence of contact dermatitis, the aim of the study is to determine the impact of allergy to cosmetics. METHODS: From January 1998 to December 1999, 819 patients were subjected to epicutaneous tests for suspicion of allergic contact dermatitis (ACD). The results were analysed retrospectively in the total population and in that of children under 16: the interest has been focused on cosmetics. RESULTS: 297 patients (36.3%) with ACD to 1 or more cosmetic ingredients have been detected. 34 (48.6%) out of 70 children patch tested showed an ACD: cosmetics represented the first cause. CONCLUSION: For two decades, the incidence of ACD has been rising; two main reasons can be put forward: a rising product consumption and a more exhaustive allergen research in patch testing. PMID- 12077542 TI - A case of early congenital syphilis. AB - A case of early congenital syphilis is reported in order to emphasize that syphilis is still present nowadays and that detection of syphilis in pregnant women is primordial for the prevention of this disease. PMID- 12077543 TI - Cutaneous Mycobacterium chelonae infection extending distally in a hemodialysed patient. AB - We report a case of a cutaneous Mycobacterium chelonae infection on the dorsum of the forearm of a patient undergoing chronic hemodialysis. The infection showed a linear and distal extension. This unusual distal dissemination was apparently secondary to a venous reflux, a circulatory repercussion of the patient's arteriovenous fistula. PMID- 12077545 TI - Bowen's disease of the nail. AB - We present a case of Bowen's disease of the nail bed in a 60-year-old male. This case should remind us to biopsy any chronic lesion of the nail in order to reduce the delay in diagnosis. Indeed Bowen's disease of the nail unit often presents as a clinically misleading 'benign' lesion. Early diagnosis is important as this precancerous lesion appears to have a more aggressive behavior when it affects the nail unit. Bowen's disease of the nail is therefore considered as a squamous cell carcinoma, and both entities are grouped under the term 'epidermoid carcinoma'. Surgical excision is the first treatment option for those tumors without bone involvement. Our patient was treated with Mohs' micrographic surgery which is considered the ideal surgical treatment as it ensures a complete eradication of the tumor in over 90% of the cases but also maximally preserves normal tissue and function. This case illustrates the challenge of Mohs' surgery on the nail unit due to its particular anatomy and histology. PMID- 12077544 TI - A double-blind placebo-controlled study of ketoconazole + desonide gel combination in the treatment of facial seborrheic dermatitis. AB - BACKGROUND: The pathobiology of seborrheic dermatitis is rooted in a peculiar inflammatory reaction to Malassezia spp. Both topical corticosteroids and antifungals are routinely used with success, either singly or in combination. OBJECTIVE: To assess the effect of a new combination therapy in this condition. METHODS: Eighteen patients with facial seborrheic dermatitis were treated in a pilot double-blind trial with an anhydrous gel containing the combination of 2% ketoconazole and 0.05% desonide (n = 9) or the unmedicated gel (n = 9). The products were applied once daily for 3 consecutive weeks. Disease severity was assessed at the start and weekly during treatment, as well as 2, 4 and 8 weeks after the end of treatment or at the time of recurrence. The overall clinical assessments were performed using a visual analogue scale. Objective measurements included squamometry X after harvesting the stratum corneum and the erythema index yielded by narrow-band reflectance spectrophotometry. Data were evaluated in an intent-to-treat analysis. RESULTS: The combination therapy yielded reductions in the overall clinical symptom severity of 49, 84 and 92% after 1, 2 and 3 weeks of treatment, respectively. The corresponding improvements with the placebo gel were significantly smaller (p < 0.01), reaching 23, 29 and 42% at successive 1-week intervals. At the end of the 3-week treatment, the clinical signs had completely subsided in 6/9 patients treated with the medicated gel and in only 1/9 patients using the placebo gel. Thus, the clinical response rate was significantly faster (p < 0.01) with the combination therapy than with the vehicle. Differences between the treatment groups using objective noninvasive assessments were also significantly (p < 0.01) in favor of the combination therapy. The squamometry X measurements showed 47, 67 and 74% improvements with the combination gel after 1, 2 and 3 weeks, respectively. The corresponding values with the placebo gel were 34, 46 and 53%, respectively. The erythema index was almost completely reduced to normal at week 2 of treatment in contrast with about 50% reduction with the unmedicated vehicle. None of the patients reported any side effects. CONCLUSION: The combination of ketoconazole with desonide in an anhydrous gel is well tolerated and proves to be significantly more effective than the vehicle for treating facial seborrheic dermatitis. The fast response to treatment and the limited additional improvement between weeks 2 and 3 of the treatment suggest that this combination product may be efficacious with once daily applications for 2 weeks. These data call for expanded studies comparing each single compound, the vehicle and the combination therapy. PMID- 12077546 TI - Diffuse plane normolipaemic xanthomatosis in a patient with chronic lymphatic leukaemia and monoclonal gammopathy. AB - Diffuse plane normolipaemic xanthomatosis is a well-defined uncommon condition characterized by yellow-orange plaques involving the eyelids, neck, upper trunk, buttocks and flexural folds. Over half of the reported cases are associated with lymphoproliferative disorders. The condition may arise as a result of perivascular deposition of lipoprotein-immunoglobulin complexes. We present a female patient with clear clinical features of diffuse plane xanthomatosis associated with chronic lymphatic leukaemia and monoclonal gammopathy. In this patient, we found evidence of complement consumption supporting the hypothesis that complexes of lipoprotein and paraprotein are formed. Lipid electrophoresis of fasting serum revealed the presence of a faint precipitate at the application slot, also indicative of the presence of protein-lipid complexes. PMID- 12077547 TI - Gamma-probe-directed lymphatic mapping and sentinel lymphadenectomy in primary cutaneous melanoma. AB - BACKGROUND: Radiotracer and blue-dye lymphatic mapping is a recommended combined method to guide sentinel lymphadenectomy and full regional lymph node dissection in selected patients with cutaneous melanoma. OBJECTIVE: To evaluate the diagnostic accuracy and the prognostic value of gamma-probe-directed lymphatic mapping in cutaneous melanomas. METHODS: Sixty-five stage I and II melanoma patients underwent gamma-probe-directed lymphatic mapping. Sentinel lymph nodes were studied by both conventional and immunohistochemical stainings. The median follow-up was 11 months. RESULTS: Sensitivities of preoperative and intraoperative sentinel lymph node detection were 100 and 98%, respectively. Only 1 failure of detection and 1 missed same-basin metastasis were experienced in the axillary and cervical areas, respectively. Eleven patients (16.9%) had sentinel node metastases leading to adjuvant therapy. CONCLUSION: Gamma-probe-directed lymphatic mapping is useful for staging melanoma. However, in the expectation of a more specific identification of the sentinel lymph node, the standard protocol remains recommended for exploring the axillary and cervical areas. The histological examination supported in some cases by immunohistochemistry remains mandatory in all cases. PMID- 12077548 TI - Multiple HPV-positive basal cell carcinomas on the abdomen in a young pregnant woman. AB - Basal cell carcinoma (BCC) is the most common malignant skin tumour. In the last few years, the incidence of multiple BCC has also increased in young patients. We describe the clinical case of a young 29-year-old woman who developed 7 BCC on her abdomen during her first pregnancy and 4 other similar tumours 2 years later during her second pregnancy, all located on the abdomen. Polymerase chain reaction revealed the presence of numerous human papillomavirus DNA sequences. To our knowledge, such a clinical presentation has not been previously reported. Different physiopathological considerations are discussed. PMID- 12077549 TI - Thalidomide-induced morbilliform rash: diagnosis and continuation of therapy, premedicated with methylprednisolone. AB - In the past few years, thalidomide is experiencing a revival in many different fields as a last therapeutic option, because of the potential severe adverse drug reactions (ADR) induced by this drug (teratogenicity, peripheral neuropathy and sedation). Taking into consideration the increased use, we should focus on the prevalence of these and other ADR as well on their management. We describe a patient with multiple myeloma who presented with a morbilliform rash induced by thalidomide. Reintroduction of the drug confirmed the diagnosis. Nevertheless we continued the thalidomide treatment, although combined with methylprednisolone 64 mg. There was no recurrence of the cutaneous drug reaction (CDR). To the best of our knowledge, this is the first case showing that systemic steroids can help to 'treat through' a thalidomide CDR. PMID- 12077550 TI - Differential role of bFGF and VEGF for vasculogenesis. AB - Primary vascular plexus originate from angioblasts through a process called vasculogenesis. The precise role of basic fibroblast growth factor (bFGF) and the vascular endothelial growth factor (VEGF) are both suggested as key regulators in vasculogenesis is still unclear. This crucial aspect was investigated by using time lapse observation of in vitro generated embryonic stem (ES) cell-derived endothelial structures which were recognizable by using the platelet cell adhesion molecule (PECAM-1) driven endothelial-specific expression of the live reporter gene enhanced green fluorescent protein (EGFP). In serum free conditions VEGF led to improved survival of angioblasts and to the formation of primitive endothelial tubes whereas bFGF alone increased their survival. Our study suggests that the complex process of vasculogenesis can be driven by VEGF alone but not by bFGF. PMID- 12077551 TI - Cyclooxygenases in the collecting duct of neonatal rabbit kidney. AB - COX 1 and 2 expression during the terminal phase of kidney development is poorly understood. To obtain information about this process we followed the primary appearance of cyclooxygenases in collecting duct (CD) epithelia of the neonatal rabbit kidney with immunohistochemical and two dimensional electrophoretical methods. In the fully embryonic cortical zone immunohistochemical expression of COX 1 is seen in all cells of the CD ampulla, while COX 2 is lacking within the nephron inducer. Within the matured cortical collecting duct (CCD) COX 1 and 2 immunoreactivity could not be detected. In contrast, a heterogeneous expression profile for COX 1 and 2 is found in the outer medullary CD (OMCD), since not all cells showed immunohistochemical labeling. Within the inner medullary CD (IMCD) nearly all cells express both cyclooxygenases. As revealed by western blot experiments generated embryonic CD epithelia in perfusion culture demonstrate high COX 1 presence at the begin of culture, while COX 2 is found to a minor degree. From day 3 until day 14 continuous levels of COX 1 and 2 expression are detected. Administration of 1 x 10(-7) mmol/l aldosterone does not influence COX expression, while application of 100 mmol/l NaCl increases COX 2 fourfold. The upregulation of COX 2 by a chronic NaCl load in embryonic epithelia suggests in part a constitutive and in part a facultative expression during CD cell differentiation. PMID- 12077552 TI - Ontogeny of purinergic receptor-regulated Ca2+ signaling in mouse cortical collecting duct epithelium. AB - Changes in ATP-induced increase in [Ca2+](i) during collecting duct ontogeny were studied in primary monolayer cultures of mouse ureteric bud (UB) and cortical collecting duct (CCD) cells by Fura-PE3 fluorescence ratio imaging. In UB (embryonic day E14 and postnatal day P1) the ATP-stimulated increase (EC(50) approximately 1 microM) in fluorescence ratio (DeltaR(ATP)) was independent of extracellular Ca2+ and insensitive to the P2 purinoceptor-antagonist suramin (1 mM). From day P7 onward when CCD morphogenesis had been completed DeltaR(ATP) increased and became dependent on extracellular Ca2+. This ATP-stimulated Ca2+ entry into CCD cells was non-capacitative and suramin (1 mM)-insensitive, but sensitive to nifedipine (30 microM) and enhanced by Bay K8644 (15 microM), a blocker and an agonist of L-type Ca2+ channels, respectively. Quantitative RT-PCR demonstrated similar mRNA expression of L-type Ca2+ channel alpha1-subunit, P2Y(1), P2Y(2), and P2X(4b) purinoceptors in UB and CCD monolayers while the abundance of P2X(4) mRNA increased with CCD morphogenesis. In conclusion, both embryonic and postnatal cells express probably P2Y(2)-stimulated Ca2+ release from intracellular stores. With development, the CCD epithelium acquires ATP stimulated Ca2+ entry via L-type Ca2+ channels. This pathway might by mediated by the increasing expression of P2X(4)-receptors resulting in an increasing ATP dependent membrane depolarization and activation of L-type Ca2+ channels. PMID- 12077554 TI - Characterisation of the calcium paradox in the isolated perfused pigeon heart: protection by hypothermia, acidosis and alkalosis. AB - The aim of the present investigation was to examine the conditions inducing a calcium paradox in the isolated perfused pigeon heart. Loss of mechanical and electrical activity, creatine phosphokinase and total protein release were used to define cell damage. Perfusion was performed at 36, 38, 40 and 42 degrees C and calcium deprivation lasted 5, 10, 20 or 40 min. At low temperatures even prolonged calcium depletion failed to induce a calcium paradox. After a 40 min calcium depletion at normal body temperature (42 degrees C) ventricular activity ceased and a major contraction occurred followed by an increase in resting tension. During the 20-min reperfusion period the release of creatine phosphokinase was 267.18 +/- 0.8 IU/g of dry wt and the total amount of protein loss was 109.3 +/- 1.0 mg/g of dry wt, while lower temperatures resulted in a decreased loss of protein and creatine phosphokinase. Using two different Tyrode's perfusion buffers instead of normal bicarbonate ones, a protection of the pigeon heart against the induction of this phenomenon was observed. Furthermore, acidosis as well as alkalosis protected the heart as estimated by the significant recovery of electromechanical activity, and the quite low total protein and creatine phosphokinase losses. The results of this study suggest that the basic mechanisms and damaging effects of calcium overloading are common in mammalian and pigeon hearts. PMID- 12077553 TI - Purinergic receptors have different effects in rat exocrine pancreas. Calcium signals monitored by fura-2 using confocal microscopy. AB - Pancreatic ducts have several types of purinergic P2 receptors, however, nothing is known about P2 receptors in acini. The aim was to establish whether acini express functional P2 receptors coupled to intracellular Ca2+ signals and to measure the signals ratiometrically in a confocal laser scanning microscope. Rat acini and ducts were loaded with various Ca2+ sensing fluorophores (Fluo-4, Fura Red, Calcium Green-1, Indo-1 and Fura-2). Only Fura-2 loaded evenly into acinar clusters and ducts and UV laser excitation at 351 and 364 nm gave signals showing opposite sensitivity to Ca2+ concentration changes. About 15% of acini or single cells within acini responded to ATP or UTP, yet all responded to carbachol. In contrast, all ducts responded to ATP/UTP. ATP stimulated uptake of a fluorescent anion, luciferin, but again only in a few acinar cells. RT-PCR analysis on acini samples collected from whole pancreas suspensions revealed transcripts for P2Y(2), P2Y(4) and P2X(1), P2X(4) receptors. The low number of functional P2 receptors in acini might be related to the fact that they release ATP. Thereby acini would avoid autocrine stimulation and initiation of autodigestive processes, such as occurs in pancreatitis. PMID- 12077555 TI - Effects of moderate and deep hypothermia on Ca2+ signaling in rat ventricular myocytes. AB - BACKGROUND/AIMS: We investigated whether the degree of hypothermia determines the impairment in cardiac muscle function upon rewarming and whether the sarcoplasmic reticulum Ca2+ release channel, RyR(2), contributes to hypothermia-induced changes in myoplasmic [Ca2+]. METHODS: Tension measurements using rat papillary muscle and calcium transients (Fluorescent Ca2+ indicator Fura 2-AM) in rat ventricular myocytes were compared during deep (10 degrees C-16 degrees C) and moderate hypothermic (28 degrees C) myocardial temperatures. In a second experiment, myocytes were pretreated with dantrolene, an RyR(2) antagonist; calcium transients were determined at control temperatures (32 degrees C), 16 degrees C, and upon rewarming (32 degrees C). RESULTS: Papillary muscle contractility and myocyte calcium transients were significantly reduced during and after rewarming from 16 degrees C. At 28 degrees C, papillary muscle isometric tension was potentiated and calcium transients were unaffected. After rewarming from 28 degrees C, excitation-contraction coupling was maintained as isometric tension returned to 90% of control values. After rewarming from 16 degrees C, myocytes pretreated with dantrolene had return of calcium transients to 89% of control values while myocytes not treated with dantrolene recovered to only 50% of their control values. CONCLUSION: We conclude that deep hypothermia, as opposed to moderate hypothermia of the myocardium, disrupts excitation contraction coupling at cellular and tissue levels. Our finding of preserved calcium transients in dantrolene-pretreated myocytes exposed to deep hypothermia suggests a potential role for the RyR(2) channel in post-hypothermia reductions in cardiac function. PMID- 12077556 TI - Phosphorylation of c-Jun N-terminal kinase in human hepatoblastoma cells is transiently increased by cold exposure and further enhanced by subsequent warm incubation of the cells. AB - During a cold preservation and reperfusion process of organs, cells are exposed to two major stresses, i.e. changes in oxygen concentration and temperature. c Jun N-terminal kinase (JNK) /stress-activated protein kinase is activated by various stresses through its phosphorylation. Although hypoxia and subsequent reoxygenation is known to activate JNK, little is known about effects of hypothermia and subsequent rewarming on JNK activation. Thus, we investigated the activation of JNK in human hepatoblastoma (HepG2) cells exposed to a temperature of 5 degrees C and in those rewarmed at 37 degrees C. Western blot analysis using an anti-phospho-JNK antibody revealed that p54 JNK was transiently phosphorylated in cold-stressed cells. In addition, the phosphorylation of p54 JNK was further increased by rewarming of the cells. Since translational and transcriptional abilities were markedly reduced in the cold-stressed cells, effects of translation and transcription inhibitors on the phosphorylation of p54 JNK were determined. Cycloheximide, but not actinomycin D, increased the phosphorylation of p54 JNK in HepG2 cells. These results suggest that hypothermia alone transiently increases the p54 JNK phosphorylation possibly through reduction of protein synthesis and that rewarming after hypothermia stimulates the phosphorylation of p54 JNK. PMID- 12077557 TI - Effect of zinc ions on caffeine-induced contracture in vascular smooth muscle and skeletal muscle of rat. AB - The effect of zinc ions on caffeine-induced contracture in vascular smooth muscle and skeletal muscle of rat was studied. In aortic strips, caffeine contracture was depressed by Zn2+ in a dose-dependent manner. Moreover, the extent of the depression of caffeine contracture in the Zn2+ loaded smooth muscle increased with repetitive caffeine exposures. For instance, in the preparations perfused with a medium containing 100 microM [Zn2+] for 15 or 30 min, the first caffeine contractures were similarly depressed to about 60% of the control. However, the subsequent caffeine exposure at 15 min interval could not evoke any contracture. In this study this feature is referred to as activation dependence of the Zn2+ effect. In small bundles of soleus muscle 2n100 microM [Zn2+] similarly caused a depression of caffeine contracture, and the activation dependence also was evident. However, an evident potentiation of caffeine contracture was seen in the preparations loaded with lower [Zn2+] such as 0.5 microM, indicating that the effect of Zn2+ on caffeine contracture of skeletal muscle was somewhat different from that seen in smooth muscle. By observing how the depression effect depends on the intervals between caffeine exposures as well as on caffeine concentrations, it is indicated that the activation dependence of the Zn2+ effect, at least in skeletal muscle, may not be explained by depletion of intracellular Ca2+ stores alone. The possible mechanism for that caffeine contracture of smooth muscle and skeletal muscle was differentially affected by Zn2+ ions is discussed. PMID- 12077558 TI - DNA interacts with Bacillus subtilis mechano-sensitive channels in native membrane patches. AB - Much recent evidence indicates that systems devoted to the transmembrane transport of proteins and/or genetic material in bacteria comprise proteins capable of forming large pores as a key element. In several cases these pores have been observed in electrophysiological experiments after purification and reconstitution of the proteins in artificial bilayers. A comparison of their properties with those of large mechanosensitive channels observed by patch clamping bacterial proto- or spheroplasts suggests that the latter may be formed by such transport machines. In support of this hypothesis, this paper reports that the properties of high-conductance channels in the membrane of Bacillus subtilis are altered by DNA through specific interactions. Thus, the previously demonstrated interaction between DNA and the same channels reconstituted in planar bilayers, which in that system results in the transmembrane translocation of the genetic material, takes place also in the native membrane. PMID- 12077559 TI - Excessive transcription of the human serum and glucocorticoid dependent kinase hSGK1 in lung fibrosis. AB - The excessive matrix deposition in lung fibrosis is thought to be due to enhanced formation and activity of TGFbeta1, which stimulates synthesis and inhibits degradation of matrix proteins. The cellular mechanisms triggered by TGFbeta1 are still incompletely understood. Recently, a novel transcriptional target of TGFbeta1 has been identified, i.e. the human serum and glucocorticoid dependent kinase hSGK1. The present study has been performed to explore whether TGFbeta1 stimulates hSGK1 transcription in lung fibroblasts and whether lung fibrosis is associated with enhanced hSGK1 expression. As evident from Northern Blotting, TGFbeta1 strongly upregulates hSGK1 in human lung fibroblasts, an effect partially reversed by p38-kinase inhibitor SB203580. In situ hybridization experiments reveal that in intact lung tissue hSGK1 is expressed in single type II alveolar pneumocytes and macrophages. In contrast, in fibrotic lung tissue a dramatic upregulation of hSGK1 mRNA as well as a strong expression of hSGK1 protein is observed in epithelial cells and interstitial cells comprising macrophages and fibroblasts. In conclusion, in lung fibrosis, the serine/threonine kinase hSGK1 is upregulated, an effect at least partially accounted for by TGFbeta1. The full effect of TGFbeta1 requires the activation of p38 kinase. PMID- 12077560 TI - Fatty acid and triacylglycerol utilisation by perfused rat spleen: differential metabolic fate and the effect of endotoxin. AB - BACKGROUND/AIMS: The spleen uses significant amounts of non-esterified fatty acid (NEFA) and chylomicron (CM)-derived triacylglycerol (TAG); however, its utilisation of very-low-density lipoprotein triacylglycerol (VLDL-TAG), and the effect of endotoxin on lipid assimilation and fate are unknown. This study aims to define spleen utilisation of esterified and non-esterified fatty acid in quiescent and endotoxin-stimulated states. METHODS: Rat spleens were perfused for 2h with glucose (11mM) and either NEFA (oleate; 0.4mM fatty acid), rat CM (0.4mM TAG), or rat VLDL (0.4mM TAG). Lipid oxidation and tissue lipid deposition were measured. RESULTS: Total utilisation of oleate and VLDL-TAG were not significantly different. Utilisation of CM was 2-3 fold greater; however oxidation of CM-TAG (5.5nmols FA/min/gram spleen) was not significantly different from either NEFA or VLDL-TAG. More TAG (CM and VLDL) was deposited as tissue lipid compared to oxidation than NEFA; the pattern of intracellular lipid accumulation of TAG and NEFA also differed. Endotoxin (LPS) did not affect the proportion of lipid oxidised and accumulated or on TAG tissue lipid distribution but it increased the proportion of oleate recovered as tissue TAG and cholesterol ester. Perfusion with CM-TAG significantly decreased heparin-releasable lipoprotein lipase (LPL) activity compared with oleate or VLDL-perfused spleens. LPS increased LPL activity 5-fold in oleate-perfused spleens, but had no effect on VLDL-perfused spleens. CONCLUSION: VLDL-TAG is utilised by spleen to a similar extent as NEFA, but its pattern of metabolic fate resembles that of CM-TAG. Endotoxin affects the pattern of NEFA deposition in tissue lipids but has no significant effect on VLDL utilisation by the spleen. PMID- 12077561 TI - LLC-PK(1) cells maintained in a new perfusion cell culture system exhibit an improved oxidative metabolism. AB - Cultured renal proximal tubule cells dedifferentiate from an oxidative metabolism to high rates of glycolysis over time. There are many reasons why cells in culture dedifferentiate, not least being a lack of homogenous nutrient supply and poor oxygenation. To this end we have developed a new cell culture device (EpiFlow), which combines continuous perfusion of medium with continuous oxygenation of cells grown on microporous supports. LLC-PK(1) cells cultured under EpiFlow conditions were compared with the same cells grown under conventional static conditions. EpiFlow maintained cells exhibited an improved oxidative metabolism as evidenced by 1) a decreased activity of glycolytic enzymes, 2) an increase in the activity of mitochondrial phosphate-dependent glutaminase, 3) an increase in cellular ATP content, and 4) an improved morphology (increased cell height, mitochondrial density and an increased number and height of microvilli). In addition, LLC-PK(1) cells maintained under perfusion conditions exhibited an increased sensitivity to the respiratory chain blocker antimycin A as assayed by mitochondrial membrane potential (JC-1). We conclude that LLC-PK(1) cells maintained under EpiFlow conditions develop an improved oxidative metabolism that is more comparable to the in vivo situation. PMID- 12077562 TI - Lipid-lowering drugs: are adverse effects predictable and reversible? AB - Most of the currently available statins and fibric acid derivatives have been implicated in causing complications either in monotherapy or in combination therapy. Adverse events occur more often with the statins that are metabolized via the CYP enzyme system and its 3A4, 2C9 or 2C19 paths. All compounds interfering with the same cytochrome system may either impair or enhance the elimination of statins. Other factors predisposing to adverse effects are age, female sex, renal insufficiency, electrolyte disturbances, infections and trauma. Complications chiefly concern the hepatic function, skeletal muscles and peripheral nerves. The major adverse effect is myopathy, up to rhabdomyolysis with ensuing acute renal insufficiency. Fibrates bind to peroxisome proliferator activated nuclear receptors alpha, with subsequent stimulation of fatty acid oxidation and reduction in the rate of hepatic lipid generation. Fibrates are associated with a number of adverse effects, including liver enzyme elevations, gastrointestinal side effects and rhabdomyolysis. The combination of statins with fibrates may cause serious complications and should be avoided when possible. In order to prevent or minimize adverse clinical outcomes, patients should be closely monitored and informed of the most common symptoms. PMID- 12077563 TI - Association between electrocardiographic pattern and level of C-reactive protein in patients with first anterior wall acute myocardial infarction. AB - BACKGROUND: Studies of prognostic markers after first anterior wall acute myocardial infarction (AMI) have reported an association between persistent ST segment elevation on the electrocardiogram (ECG) and increased levels of C reactive protein (CRP) and worse outcome. The aim of the present study was to determine the nature of the relationship between these two factors. METHODS: Twenty-three consecutive patients admitted with AMI underwent 12-lead ECG on days 1, 2, 3 and 7 and were classified into two groups according to the findings on each day's recording: pattern A - isoelectric ST segment, or pattern B - ST segment elevation with or without a negative T wave. Serum CRP levels measured on the same days were also compared between the groups. RESULTS: The number of patients classified as pattern A increased over time, from 8 on day 1 after AMI to 11 on day 2, 12 on day 3 and 14 on day 7, with a corresponding decrease in pattern B patients. Most of the patients with pattern B on day 1 showed a persistent pattern on day 7. For each day studied, the pattern A and pattern B patients showed a statistically significant difference in serum CRP levels on the second and third days after AMI, respectively. Values for the day 7 groups were: 3.1 +/- 3.4 versus 9.6 +/- 3.8 mg% (p = 0.001) (day 2) and 3.1 +/- 1.2 versus 7.9 +/- 3.6 mg% (p = 0.0005) (day 3). CONCLUSION: There was a significant association between high CRP levels on the second and third day following first AMI and ECG pattern B (ST elevation). This conclusion strengthens our earlier findings that pattern B is predictive of poor outcome in this patient population. PMID- 12077564 TI - Sexual activity of women and men one year before a first acute myocardial infarction. AB - A decrease in sexual activity after an acute myocardial infarction (AMI) for both women and men has been reported with one study suggesting that this reduction in sexual activity may precede rather than proceed the AMI. The present study compared the sexual activity of women and men before AMI to the sexual activity of a normative community sample of women and men, to examine whether the above reduction in sexual activity is especially characteristic of women and men who later incur an AMI. This study also investigated the association between selected medical and sociodemographic variables and sexual inactivity of women and men before an AMI. During an interview before discharge,138 women and 760 men who were hospitalized due to a first AMI responded to a question regarding the frequency of their sexual activity 1 year before the AMI. Their sociodemographic and medical background was obtained from the interview and the medical charts. When compared to a normative sample, only women reported significantly less sexual activity during the year before their AMI. These women were also found to be at a disadvantage when compared to men on many of the sociodemographic and medical variables shown to contribute to sexual inactivity for both men and women. However, the higher percent of sexual inactivity for women during the year before AMI may not only be due to women's higher morbidity and lower sociodemographic status. Other variables not included in this study, but associated with gender, could account for this result. PMID- 12077565 TI - Predictors of clinical recurrence after successful electrical cardioversion of chronic persistent atrial fibrillation: clinical and electrophysiological observations. AB - Recurrence of atrial fibrillation (AF) after electrical cardioversion of chronic AF is not uncommon. However, it remains unclear which parameter(s) predict clinical recurrence. To assess the potential predictors of clinical recurrence after successful electrical cardioversion, we analyzed clinical, echocardiographic and electrophysiologic parameters in 36 patients (age 63 +/- 11 years; 26 males, 10 females) with chronic persistent AF lasting more than 3 months. The dimensions of the left atrium and left ventricular end diastole and end systole were measured by echocardiography. The P wave characteristics from the surface 12-lead electrocardiogram (ECG) were studied in sinus rhythm. Atrial local activations were studied by biatrial basket electrode mapping in AF. With a mean of 7 +/- 2 months of follow-up, 17 (47%) patients had AF recurrence despite multiple antiarrhythmic drug therapy. None of the clinical or echocardiographic parameters were relevant to the recurrence. However, among the surface ECG and intraatrial electrophysiological parameters, the mean P wave duration was the only independent predictor of the risk of clinical recurrence after successful electrical cardioversion. The sensitivity and predictive accuracy of recurrence were 82 and 70%, respectively, when the mean P wave duration was more than 125 ms. PMID- 12077566 TI - Interactions of esmolol and adenosine in atrioventricular nodal-dependent supraventricular tachycardia: implication for the cellular mechanisms of adenosine. AB - INTRODUCTION: Cellular mechanisms of adenosine include a direct effect on the activation of the adenosine-sensitive potassium current (I(K,Ado)) and an indirect effect on antagonism of catecholamine-stimulated adenylate cyclase activity. However, previous studies evaluating the influence of catecholamine activity on the electrophysiologic effects of adenosine have yielded conflicting results. We tested the hypotheses that if adenosine exerts its atrioventricular (AV) nodal blocking effects directly by activating the I(K,Ado) potassium current, rather than indirectly by reversing the catecholamine effects, then pretreatment with beta-adrenergic blockade would not potentiate the effects of adenosine in terminating AV nodal-dependent supraventricular tachycardia (SVT). METHODS AND RESULTS: During sustained AV nodal reentrant tachycardia (AVNRT) or AV reentrant tachycardia (AVRT) in 28 patients, adenosine was rapidly injected in incremental doses of 1.5, 3, 6, 9, 12 and 18 mg to determine the lowest effective dose required for tachycardia termination before and immediately after the end of esmolol infusion. Esmolol infusion was started with loading doses of 500 /kg/min for 1 min and 150 microg/kg/min for 4 min, followed by a maintenance infusion of 50-100 microg/kg/min. Esmolol infusion was continued until the tachycardia was terminated or the maximal dose of 100 mg was reached. Adenosine was effective in terminating SVT in all 28 patients with a mean lowest effective dose of 96 +/- 54 microg/kg before esmolol. During esmolol infusion, tachycardia was reproducibly terminated in 8 patients (6 with AVNRT, 2 with AVRT) with a mean dose of 67 +/- 23 mg. In the other 20 patients with persistent tachycardia after 100 mg of esmolol infusion, the lowest effective dose of adenosine could be determined in 19 patients. In the remaining patient with AVRT, the maximal dose of adenosine (18 mg) was unable to terminate the tachycardia immediately after the end of esmolol infusion. In these 19 patients, esmolol infusion caused significant lengthening of the tachycardia cycle length from 338 +/- 36 to 372 +/- 51 ms (p < 0.0001) and reduction of the mean arterial blood pressure from 96 +/- 15 to 88 +/ 18 mm Hg (p = 0.034). Compared to the dosage that was determined before esmolol infusion, the lowest effective dose of adenosine remained the same in 13 patients after the end of esmolol infusion, whereas the dose increased in 5 and decreased in 1 patient. The mean lowest effective dose of adenosine was not significantly different before (98 +/- 54 microg/kg) and immediately after (115 +/- 56 microg/kg) the end of esmolol infusion (p = 0.054). CONCLUSIONS: Intravenous esmolol infusion (up to 100 mg total dose) usually fails to terminate AV nodal dependent SVT. In the esmolol-resistant tachycardia, esmolol pretreatment does not produce a positive synergistic effect on the efficacy of adenosine-induced termination of SVT. Therefore, in this tachycardia adenosine may exert its effects on AV nodal conduction directly by activation of the I(K,Ado) potassium current, rather than by antagonizing the beta-adrenergic system. PMID- 12077567 TI - Differential effects of adenosine on antegrade fast pathway, antegrade slow pathway, and retrograde fast pathway in atrioventricular nodal reentry. AB - Adenosine has a potent negative dromotropic effect. However, comparative effects of adenosine on the three pathways of atrioventricular (AV) nodal reentry remain unclear. In this study, we sought to determine the effects of adenosine on the antegrade fast, antegrade slow, and retrograde fast pathway conduction in patients with AV nodal reentrant tachycardia (AVNRT). Twenty patients with common slow-fast AVNRT (mean cycle length 360 +/- 49 ms) were studied. The effects of adenosine on the antegrade slow pathway and on the retrograde fast pathway conduction were determined during sustained AVNRT and constant right ventricular pacing at identical cycle lengths (mean 360 +/- 49 ms), respectively. Incremental doses of adenosine were rapidly administered: initial dose of 0.5 mg, followed by stepwise increases of 0.5 or 1.0 mg given at 5-min intervals until termination of AVNRT or second-degree ventriculoatrial block occurred. After the antegrade slow pathway conduction was selectively and completely ablated by radiofrequency catheter ablation, the effect of adenosine on the antegrade fast pathway conduction was evaluated. The dose-response curve of adenosine and the dose of adenosine required to produce AV or ventriculoatrial block among the representative three conduction pathways were compared. The dose-response curve for the effect of adenosine on the antegrade fast pathway lies to the left and upward to that of the effect of adenosine on the antegrade slow pathway which in turn lies to the left and upward to that of the retrograde fast pathway. The mean dose of adenosine required to produce conduction block at antegrade fast, antegrade slow, and retrograde fast pathways were 1.4 +/- 0.5, 4.2 +/- 1.6, and 8.5 +/- 2.6 mg, respectively (p < 0.01). Adenosine has a differential potency to depress antegrade fast, antegrade slow, and retrograde fast pathway conduction in patients with AVNRT. The depressant effect of adenosine on the antegrade fast pathway is more potent than that on the antegrade slow pathway which in turn is more potent than that on the retrograde fast pathway conduction. PMID- 12077568 TI - Effect of balloon mitral valvoplasty during pregnancy on childhood development. AB - In order to study the long-term effects of balloon mitral valvoplasty (BMV) on childhood development, we followed up closely 20 mothers with severe mitral stenosis who were treated with BMV during pregnancy. Their offspring aged 63 +/- 39 (range 6-98) months were also regularly assessed using standard developmental charts and relevant laboratory tests. They were all found to have maintained normal growth, development, and speech for their appropriate ages. Developmental milestones were normal for their ages and comparable with those of their siblings. The laboratory tests revealed no abnormalities. Two losses occurred during the early course of follow-up, at the ages of 4 and 37 weeks, respectively. One of the pregnant patients suffered a stillbirth. PMID- 12077569 TI - Prehospital delay in individuals with acute coronary disease: concordance of medical records and follow-up phone interviews. AB - Patient-associated delay in seeking medical care in persons with acute coronary disease is receiving increasing importance given the time-dependent benefits associated with myocardial reperfusion therapies. We examined the extent of concordance between self-reported information about prehospital delay provided by patients to hospital staff at the time of hospitalization for coronary disease compared with information obtained from a telephone interview approximately 2 months following hospital discharge. The sample included 316 patients with acute myocardial infarction or unstable angina at 43 hospitals who had delay time information available from both data sources. The extent of agreement between the medical record and telephone accounts of delay was 47% in the total study sample, 53% in patients with acute myocardial infarction, and 40% in patients with unstable angina. These results suggest that a telephone interview carried out several months following hospitalization for acute coronary disease may not provide sufficiently reliable information about prehospital delay. PMID- 12077570 TI - Grade III ischemia on presentation with acute myocardial infarction predicts rapid progression of necrosis and less myocardial salvage with thrombolysis. AB - We assessed the relation between baseline electrocardiographic ischemia grades and initial myocardial area at risk (AR) and final infarct size (IS) in 49 patients who had undergone (99m)Tc sestamibi single-photon emission computed tomography before and 6 +/- 1 days after thrombolysis. Patients were classed as having grade III ischemia (ST segment elevation with terminal QRS distortion, n = 19) or grade II ischemia (ST elevation but no terminal QRS distortion, n = 30). We compared AR and IS by baseline ischemia grade and treatment (adenosine vs. placebo) and assessed relations of infarction index (IS/AR ratio x100) to time to thrombolysis, baseline ischemia grade, and adenosine therapy. Time to thrombolysis was similar for grade II and grade III. For placebo- treated patients, the median AR did not differ significantly between grade II (38%) and grade III patients (46%, p = 0.47), nor did median IS (16 vs. 40%, p = 0.096), but the median infarction index was 66 vs. 90% (p = 0.006). For adenosine-treated patients, median AR (21 vs. 26%, p = 0.44), median IS (5 vs. 17%, p = 0.15), and their ratio (31 vs. 67%, p = 0.23) did not differ significantly between grade II and grade III patients. The infarction index independently related to grade III ischemia (p = 0.0121) and adenosine therapy (p = 0.045). Infarct size related to baseline ischemia grade and was reduced by adenosine treatment. Necrosis progressed slowlier with baseline grade II versus III ischemia, which could offer more time for myocardial salvage with reperfusion. PMID- 12077572 TI - Bony injuries of the shoulder. AB - The complex anatomy and functioning of the shoulder may challenge the radiologist. In this review the patterns of injury and radiographic findings for bony injuries of the shoulder are illustrated. The radiographic and cross sectional imaging findings in fractures of the clavicle, scapula, and proximal humerus and in dislocations of the sternoclavicular, acromioclavicular, and glenohumeral articulations are discussed. PMID- 12077573 TI - Dual-energy digital subtraction chest radiography: technical considerations. AB - In the evaluation of asbestos-related pulmonary and pleural abnormalities, conventional chest radiography has been shown to have a low sensitivity for the detection of lung nodules and subtle interstitial disease. Pleural plaques may simulate pulmonary nodules, and interstitial processes can be masked by adjacent pleural abnormalities. Dual-energy digital subtraction chest radiography may enable investigators to characterize asbestos-related pulmonary and pleural abnormalities with greater accuracy. "Soft-tissue" images, designed to remove pleural calcifications, may allow for better evaluation of the lung parenchyma. "Bone" images, designed to remove soft-tissue structures, may enhance the detection of pleural calcifications. In this pictorial essay we illustrate the methods, technical considerations, and limitations of dual-energy digital subtraction chest radiography performed with global subtraction weighting factors. PMID- 12077574 TI - Pulmonary interstitial emphysema. PMID- 12077575 TI - Skin cancer in organ transplant recipients: Epidemiology, pathogenesis, and management. AB - In the United States more than 100,000 people are living with solid organ transplants. The intense immunosuppressive regimens necessary for prolonged survival of allografts significantly increase the rates of both internal and cutaneous malignancies in recipients of solid organ transplants. Skin cancer is the most common cancer in patients after transplantation. Because of the early onset and high tumor burden in transplant recipients, dermatologists have significant challenges in managing the treatment of these patients. This article describes the epidemiology and clinical presentation of skin cancer during posttransplantation immunosuppression, discusses pathogenic cofactors, and reviews the optimal management for mild and severe skin cancer in transplant recipients. PMID- 12077577 TI - Prostaglandin E2 induces vasodilation and pruritus, but no protein extravasation in atopic dermatitis and controls. AB - BACKGROUND: Prostaglandin E(2) (PGE(2)) has well-established vasodilatory effects, whereas its effects on protein extravasation and its sensory effects are less clear. OBJECTIVE: Vasoactive effects of PGE(2) were correlated to its ability to evoke pain or itch in healthy volunteers and patients with atopic dermatitis (AD). METHODS: Intradermal microdialysis was used to apply PGE(2) (10( 8)-10(-4) M) via microdialysis capillaries in 8 patients and 8 controls. Large pore size membranes (3000 kd) enabled simultaneous analysis of protein extravasation. Itch and pain sensations were measured psychophysically, and superficial blood flow was measured by laser Doppler imaging. RESULTS: PGE(2) dose dependently provoked intense local vasodilation, weak pruritus, and pain, but no protein extravasation. No differences were found between patients with AD and controls for any parameter. CONCLUSION: We conclude that PGE(2) is a potent vasodilator and a weak pruritic agent in normal skin and in patients with AD, but does not provoke increased protein extravasation. PMID- 12077576 TI - Circumscribed palmar or plantar hypokeratosis: a distinctive epidermal malformation of the palms or soles. AB - BACKGROUND: Epidermal malformations of the skin include a group of heterogeneous developmental defects that result from errors in morphogenesis of the epidermis during intrauterine life. OBJECTIVE: The purpose of this study was to report the clinical and histopathologic features of a distinctive epidermal malformation involving the skin of the palms or soles. METHODS: Ten patients were included in this study. All of them showed the same clinical features that consisted of a solitary circumscribed and circular area of erythematous depressed skin on the palm or on the sole. Diagnosis was confirmed by histopathologic study. RESULTS: All patients were middle aged or elderly. Nine patients were women and one was a man. The lesions showed predilection for the skin of the thenar and hypothenar regions of the palm or the medial side of the sole. Histopathologic study demonstrated a depression of the epidermis, with a sharp stair between normal and involved skin. The epidermis covering the depression showed markedly thinner horny layer and a slightly diminished granular cell layer when compared with adjacent noninvolved skin. Keratinocytes of the squamous cell layer, granular cells, and corneocytes showed, otherwise, a normal appearance. Serial sections failed to demonstrate cornoid lamellation. CONCLUSION: On the basis of the clinical and histopathologic findings in these 10 patients, we have named this malformation circumscribed palmar or plantar hypokeratosis. This lesion seems to be a distinctive entity that has not been previously described. PMID- 12077578 TI - The persistent risk of genital tumors among men treated with psoralen plus ultraviolet A (PUVA) for psoriasis. AB - BACKGROUND: In the general population, squamous cell carcinomas (SCCs) of the male genitalia are rare. Ten years ago, we documented a significant dose dependent increase in the risk of malignant genital neoplasms among men treated with psoralen plus ultraviolet A (PUVA). Since that time, fewer cohort patients have used PUVA, and genital protection among PUVA users is likely to be frequent. OBJECTIVE: Our aim was to determine the incidence and risk factors for genital neoplasms since 1989 and risk factors for these neoplasms among patients treated with PUVA. METHODS: We conducted a prospective cohort study of 892 men first treated with PUVA in 1975-1976. RESULTS: Twenty-four men (2.7%) had 51 genital neoplasms, including 10 patients with a first tumor after May 1, 1989 (the ending date for our 1990 report). Since May 1, 1989, the incidence of invasive penile and scrotal SCCs was elevated 52.6-fold (95% confidence interval, 19.3-114.6) compared with that expected for the general white population. Multivariate models revealed the highest genital tumor risk among men with high-dose exposure to both PUVA and topical tar/ultraviolet B, with an incidence rate ratio of 4.5 (95% confidence interval, 1.3-16.1) compared with the low-dose exposure group. CONCLUSION: Although use of PUVA has decreased and genital shielding in our cohort has increased, the dose-dependent increase in the risk of genital tumors in men treated with PUVA has persisted. Particularly high risks occur among those with high-dose exposures to both PUVA and topical tar/ultraviolet B. PMID- 12077580 TI - Paraneoplastic acral vascular syndrome: epidemiologic features, clinical manifestations, and disease sequelae. AB - BACKGROUND: Acral vascular syndromes associated with malignancy have rarely been reported. OBJECTIVE: Our purpose was to assess the clinical and evolving features of paraneoplastic acral vascular syndromes. PATIENTS AND METHODS: Two cases of paraneoplastic gangrene are described and analyzed together with previously reported cases identified by a MEDLINE search. RESULTS: Among the 68 patients identified, 40 had gangrene, 16 had acrocyanosis, and 12 had Raynaud's phenomenon. The male to female ratio was 0.89; median age was 59 years. Fingers were affected in 94%. Adenocarcinomas were the predominant associated malignancies (41%), and metastases were observed in 41%. The acral vascular syndromes in 48% of the patients definitively regressed after tumor treatment. Forty-four percent of the patients died within 2 years. A favorable cutaneous outcome was obtained with prostacyclin infusions in 6 patients. CONCLUSION: A neoplastic origin of acral vascular syndrome should be considered in elderly patients, especially men, in the absence of usual causative conditions. PMID- 12077579 TI - Prospective evaluation of risk factors of cutaneous drug reactions to sulfonamides in patients with AIDS. AB - BACKGROUND: Persons with HIV infection have increased rates of drug eruptions. OBJECTIVE: Our aim was to evaluate the risk factors of drug eruptions in response to sulfonamides in patients with AIDS, using a case-control analysis. METHODS: One hundred thirty-six patients who were hospitalized for pneumocystosis or toxoplasmosis were evaluated at the onset of treatment for various risk factors, which were then compared among patients with (48, 36%) and without (88, 64%) a drug eruption. RESULTS: In multivariate analysis, high CD8(+) cell count and age less than 36 years indicated a risk of drug eruption (respective odds ratios: 3.5 [95% CI 1.6-7.8], P =.002, and 2.1 [95% CI 1-4.6], P =.06). Markers of viral replication for HIV, Epstein-Barr virus, cytomegalovirus, human herpesvirus 6, and parvovirus B19, slow acetylation phenotype or genotype, and glutathione level were not associated with a risk. Administration of corticosteroids had no preventive effect. CONCLUSIONS: Our results challenge several current concepts regarding drug eruptions by discarding a strong association with glutathione deficiency, slow acetylation, or active viral infections and by showing no preventive effect of corticosteroids. PMID- 12077581 TI - Three-dimensional analysis of a calciphylaxis plaque: clues to pathogenesis. AB - BACKGROUND: Calciphylaxis is a rare, life-threatening disorder associated with chronic renal failure, presenting with ulcerating plaques leading to death by sepsis in 60% of patients. Calcification of subcutaneous arterioles, thromboses, and extravascular calcification have been demonstrated in incisional biopsy specimens. However, the sequence of these pathologic events is unknown. OBJECTIVE: We examined a calciphylaxis plaque to document the wave of pathologic change from its center to its periphery. METHODS: A calciphylaxis plaque was excised postmortem from a female patient. It was examined histologically along 12 radii from the center of the lesion to its periphery. RESULTS: Calcification of small subcutaneous vessels was present in all histologically abnormal sections and extended further peripherally than extravascular calcification by up to 3.0 cm and further than subcutaneous thrombosis by up to 1.5 cm. CONCLUSION: Vascular mural calcification is an early and essential process in the development of a calciphylaxis plaque. PMID- 12077582 TI - Increased interleukin 10, tumor necrosis factor alpha, and interleukin 6 levels in blister fluid of toxic epidermal necrolysis. AB - BACKGROUND: Toxic epidermal necrolysis is a severe, usually drug-induced disease that shares clinical, histologic, and immunologic similarities with the severe forms of cutaneous acute graft-versus-host disease. OBJECTIVE: Our purpose was to further characterize common immune-inflammatory pathways in these skin disorders by measurement of different cytokines. METHODS: Evaluation of serum levels of interleukin 10 (IL-10), tumor necrosis factor alpha, IL-6, and soluble IL-6 receptor in the early phase of both diseases and in blister fluid of toxic epidermal necrolysis. RESULTS: Serum levels of IL-10 and IL-6 were significantly higher in patients with toxic epidermal necrolysis (P =.0001) and acute graft versus-host disease (P =.001) compared with those of blood donors. We found an increase in IL-6 levels in blister fluid and significantly higher levels of IL-10 (P =.018) and tumor necrosis factor alpha (P =.028) in blister fluid compared with serum in patients with toxic epidermal necrolysis. CONCLUSION: A similar serum cytokine profile of toxic epidermal necrolysis and acute graft-versus-host disease further emphasizes common immunologic mechanisms. The presence of inflammatory cytokines, IL-6 and tumor necrosis factor alpha, in the blister fluid of patients with toxic epidermal necrolysis is associated with significantly higher levels of IL-10, which through its down-regulatory role, may be involved in limitation of the disease extension. PMID- 12077583 TI - Effect of insulin-dependent diabetes mellitus on response to extracorporeal photopheresis in patients with Sezary syndrome. AB - BACKGROUND: Extracorporeal photopheresis (ECP) has become a primary therapy for selected forms of cutaneous T-cell lymphoma, especially Sezary syndrome. Variability in response of patients with Sezary syndrome to ECP has been reported. OBJECTIVE: Our purpose was to determine whether underlying medical conditions influence the efficacy of ECP in patients with Sezary syndrome. METHODS: We retrospectively reviewed the medical records of 55 patients with Sezary syndrome who received ECP between 1987 and 2000. Efficacy criteria included decrease in Sezary cell count, erythroderma, lymphadenopathy, organomegaly, and pruritus. RESULTS: Thirty-four patients responded well and 10 patients responded partially to ECP; 11 patients had no response. Nine patients with no response to ECP had insulin-dependent diabetes mellitus (IDDM). IDDM was documented in only 2 patients with a good response and in no patients with a partial response to ECP. CONCLUSION: Patients with Sezary syndrome and IDDM typically respond poorly to the standard ECP treatment regimen. PMID- 12077584 TI - Patient and referring provider satisfaction with teledermatology. AB - BACKGROUND: Teledermatology has become more widely used, but its impact has not been clearly elucidated. We developed a teledermatology program in response to clinical need, based on the store-and-forward approach. OBJECTIVE AND METHODS: Our purpose was to evaluate the satisfaction of patient and referring physician by using telephone interview. RESULTS: Both patients and providers considered this a useful program and would recommend it to a friend or colleague. The patients were split in their overall rating of the program and its ability to treat their skin conditions. Their greatest concern was their lack of direct contact with their dermatologist. Other common concerns were waiting time and follow-up. Privacy concerns were not commonly mentioned. Providers expressed greater satisfaction than their patients. Their greatest concerns involved the inability of the program to handle the demand. CONCLUSION: The program provided a service that was valued by both patients and providers. The most common concern was the impact on the physician-patient relationship. PMID- 12077585 TI - Topical steroids versus systemic antifungals in the treatment of chronic paronychia: an open, randomized double-blind and double dummy study. AB - BACKGROUND: The involvement of Candida in the pathogenesis of chronic paronychia (CP) has never been proven, even though this condition is commonly considered a type of Candida onychomycosis. OBJECTIVE: The purpose of this study was to compare the efficacy of systemic antifungals (itraconazole or terbinafine) with a topical corticosteroid (methylprednisolone aceponate) in the treatment of patients with CP. METHODS: The study involved 45 adult patients with CP. Medication was given in a randomized, double-blind and double dummy manner over 3 weeks. Patients were then followed for 6 weeks. Clinical and mycologic evaluations were performed at baseline, and at weeks 3 and 9. The efficacy measures included clinical and photographic evaluation. RESULTS: Of 48 nails treated with methylprednisolone aceponate, 41 were improved or cured at the end of the follow-up period. The statistical analysis showed a significant difference between the number of nails improved or cured by methylprednisolone aceponate and that of nails improved or cured with terbinafine (30 out of 57) or itraconazole (29 out of 64). Presence of Candida was not strictly linked to disease activity, and Candida eradication was associated with clinical cure in only 2 of the 18 patients who carried Candida at the beginning of the study. CONCLUSION: This study shows that topical steroids are more effective than systemic antifungals in the treatment of CP, and supports the view that CP is not a type of onychomycosis but a variety of hand dermatitis caused by environmental exposure. PMID- 12077586 TI - Congenital and infantile melanoma: review of the literature and report of an uncommon variant, pigment-synthesizing melanoma. AB - Congenital and infantile types of melanoma are uncommon conditions for which there are limited epidemiologic data. The number of reported cases is small with several ascribed etiologies. We review the literature and report the first documented case, to our knowledge, of pigment-synthesizing melanoma in an infant. Reported cases of congenital and infantile melanoma were identified and categorized on the basis of disease origin. Dermatopathologic specimens from an infant given a diagnosis of pigment-synthesizing melanoma are described. Disease arising from medium and large/giant congenital nevi was most common, whereas reports of de novo and transplacental disease were infrequent. Death of approximately 40% of patients was noted within 18 months of diagnosis. Male infants accounted for approximately 74% of cases. The most commonly affected anatomic sites were the head and neck. The prognosis for congenital and infantile melanoma is poor. The high incidence of head-and-neck involvement and male predominance for disease suggest dispositions for both anatomic disease localization and sex. PMID- 12077588 TI - Liposuction. PMID- 12077589 TI - Acne: 1991-2001. PMID- 12077587 TI - Progress in heritable skin diseases: molecular bases and clinical implications. PMID- 12077590 TI - Skin cancer awareness in suburban employees: a Hispanic perspective. AB - Skin cancer is the most common form of cancer in the United States and accounts for an estimated 1 million new cases per year. In the white population, the rate of new melanomas diagnosed yearly has more than doubled from 5/100,000 in 1973 to 12/100,000 in 1990. The current lifetime risks for white persons are estimated to be 28% to 33% for basal cell carcinoma and 7% to 11% for squamous cell carcinoma. Skin cancer is more common in sun-sensitive, fair-skinned individuals but can occur in any ethnic group, especially with exposure in tropical or sunny climates. Little is known about awareness of skin cancer, risk perception, and performance of skin self-examination (SSE) by people with skin that rarely burns. Our purpose was to evaluate skin cancer awareness, perceptions of skin cancer risk, and performance of SSE in a Hispanic versus non-Hispanic white population with similar access to health care and promotion. One hundred forty people employed by a suburban city voluntarily attended a free skin cancer screening and completed a self-administered survey. Hispanic individuals reported decreased skin sensitivity and tendency to burn. Non-Hispanic individuals believed they were at greater than average risk for skin cancer, and most Hispanics believed they were at average or below average risk. None of the 27 Hispanics reported ever being taught SSE. More non-Hispanics (32%) had performed SSE within the last year than Hispanics (15%). Regular SSE was associated with a history of skin cancer. Multimedia messages usually link skin cancer warning signs and early detection strategies with having sun-sensitive skin. People without sun sensitivity did not perceive themselves as being at risk, did not learn the warning signs of skin cancer, and did not perform SSE. Awareness of melanoma and non-melanoma skin cancer and perception of risk among Hispanics were less than among non-Hispanics, which may contribute to presentation for care at an advanced stage. PMID- 12077591 TI - Lymphedema: an immunologically vulnerable site for development of neoplasms. AB - Lymphedema is the result of accumulation of protein-rich interstitial fluid (lymph stasis) caused by a failure of lymph drainage in the face of a normal capillary filtration. Whether the origin is congenital or acquired from infection, radiation, trauma, or surgery, chronic lymph stasis impairs local immune surveillance by disrupting trafficking of the immunocompetent cells in the lymphedematous district and stimulates vicarious angiogenesis by promoting development of a collateral lymphatic and hematic network in the lymphedematous district. When the local mechanisms of immune surveillance begin to fail, the lymphedematous region becomes an immunologically vulnerable area, predisposed to malignancy, chiefly vascular tumors such as Stewart-Treves syndrome and Kaposi's sarcoma, because of the continual angiogenic stimulus. PMID- 12077592 TI - A footnote on the origin of fluorosteroids. PMID- 12077593 TI - Tulle or veil fabric: a versatile option for dressings. PMID- 12077594 TI - Subacute prurigo variant of bullous pemphigoid: autoantibodies show the same specificity compared with classic bullous pemphigoid. AB - We describe a 76-year-old white woman with a 6-month history of intensive pruritus and excoriated papules resembling subacute prurigo. Histopathology showed signs of chronic dermatitis, whereas findings by direct and indirect immunofluorescence microscopy were compatible with bullous pemphigoid (BP). The patient's serum contained IgG autoantibodies that recognized epitopes on both BP180 and BP230 by Western blot analysis of epidermal extracts. In addition, we found strong reactivity with recombinant NC16A, an immunodominant region of BP180 targeted in the majority of BP sera, whereas no antibodies against the keratinocyte-derived soluble BP180 ectodomain (LAD-1) or the recombinant intracellular domain of BP180 were detected. The patient's disease responded well to oral methylprednisolone and mycophenolate mofetil. Disease activity correlated with enzyme-linked immunosorbent assay reactivity of antibodies to BP180 but not with titers of antibodies to the dermoepidermal junction as determined by indirect immunofluorescence on salt-split skin. Our findings suggest that the subacute prurigo form of BP is a true variant of BP. PMID- 12077595 TI - Morphologic changes of a pigmented Spitz nevus assessed by dermoscopy. AB - Pigmented Spitz nevus may simulate cutaneous melanoma clinically and histopathologically. In an effort to characterize Spitz nevi using dermoscopy, we documented the dermoscopic features of a single pigmented Spitz nevus over a 6 month period. A 3-year-old boy had a brownish black papule, 3 mm in diameter, on the dorsum of the first finger of his left hand, clinically diagnosed as a Reed nevus. Two follow-up examinations were performed after 3 and 6 months, when the lesion finally was excised for histopathologic examination. Dermoscopically, a globular pattern was recognized during the initial examination, whereas a starburst pattern was identified 3 months later. After 6 months, a variation of the starburst pattern was still detectable. Based on our observation, the globular and the starburst patterns might be considered different morphologic expressions corresponding to the evolutionary phases of pigmented Spitz nevi. PMID- 12077596 TI - Induction of complete remission of advanced stage mycosis fungoides by allogeneic hematopoietic stem cell transplantation. AB - Advanced mycosis fungoides (MF) is incurable with conventional treatments. High dose chemoradiotherapy with autologous bone marrow transplantation has induced remissions in a small number of patients with MF, but this modality is limited by a high relapse rate. We report induction of complete remission in a 37-year-old woman with rapidly progressive stage IV MF with allogeneic stem cell transplantation (Allo SCT). She remains in continuous complete remission 2 years after transplant. Allo SCT for MF is theoretically attractive, because there is no contamination of the graft by malignant cells, and because of the possibility of graft-versus-tumor effect. Although the results in this patient are encouraging, more patients and longer follow-up are needed to define the usefulness of Allo SCT in the treatment of MF. PMID- 12077597 TI - Nail pigmentation caused by hydroxyurea: report of 9 cases. AB - We report a series of 9 patients, 6 men and 3 women, who presented nail hyperpigmentation arising between 6 and 24 months from the start of hydroxyurea therapy. The most commonly observed clinical pattern was that of longitudinal melanonychia. In only 1 patient, who was affected in all 20 nails, we observed longitudinal melanonychia, diffuse melanonychia, and hyperpigmentation of the skin. PMID- 12077598 TI - Prevalence of 52-kd and 60-kd Ro/SS-A autoantibodies in Japanese patients with polymyositis/dermatomyositis. AB - To determine the prevalence of 52-kd and 60-kd Ro/SS-A antibodies in Japanese patients with polymyositis/dermatomyositis, we examined serum samples from 61 patients with PM/DM, 10 patients with primary Sjogren's syndrome, and 25 healthy control subjects. Six serum samples possessed anti-Ro/SS-A antibodies and were positive for anti-Ro52, anti-Ro60, or both. Two reacted with both Ro52 and Ro60, and 4 reacted with Ro52 alone. The results suggest that Ro52 is the main antigen of anti-Ro/SS-A antibodies in patients with polymyositis/dermatomyositis and that its coexistence with other defined antibodies suggests the existence of a subgroup of patients with various serologic abnormalities. PMID- 12077599 TI - Psoriasiform mycosis fungoides with fatal outcome after treatment with cyclosporine. PMID- 12077600 TI - About the pathophysiology of venous leg ulceration. PMID- 12077601 TI - Topical anesthesia: clarification. PMID- 12077602 TI - Mycosis fungoides and serology for human T-cell lymphotropic virus, type 1. PMID- 12077603 TI - Pluripotency of mesenchymal stem cells derived from adult marrow. AB - We report here that cells co-purifying with mesenchymal stem cells--termed here multipotent adult progenitor cells or MAPCs--differentiate, at the single cell level, not only into mesenchymal cells, but also cells with visceral mesoderm, neuroectoderm and endoderm characteristics in vitro. When injected into an early blastocyst, single MAPCs contribute to most, if not all, somatic cell types. On transplantation into a non-irradiated host, MAPCs engraft and differentiate to the haematopoietic lineage, in addition to the epithelium of liver, lung and gut. Engraftment in the haematopoietic system as well as the gastrointestinal tract is increased when MAPCs are transplanted in a minimally irradiated host. As MAPCs proliferate extensively without obvious senescence or loss of differentiation potential, they may be an ideal cell source for therapy of inherited or degenerative diseases. PMID- 12077604 TI - TRPV3 is a calcium-permeable temperature-sensitive cation channel. AB - Transient receptor potential (TRP) proteins are cation-selective channels that function in processes as diverse as sensation and vasoregulation. Mammalian TRP channels that are gated by heat and capsaicin (>43 degrees C; TRPV1 (ref. 1)), noxious heat (>52 degrees C; TRPV2 (ref. 2)), and cooling (< 22 degrees C; TRPM8 (refs 3, 4)) have been cloned; however, little is known about the molecular determinants of temperature sensing in the range between approximately 22 degrees C and 40 degrees C. Here we have identified a member of the vanilloid channel family, human TRPV3 (hTRPV3) that is expressed in skin, tongue, dorsal root ganglion, trigeminal ganglion, spinal cord and brain. Increasing temperature from 22 degrees C to 40 degrees C in mammalian cells transfected with hTRPV3 elevated intracellular calcium by activating a nonselective cationic conductance. As in published recordings from sensory neurons, the current was steeply dependent on temperature, sensitized with repeated heating, and displayed a marked hysteresis on heating and cooling. On the basis of these properties, we propose that hTRPV3 is thermosensitive in the physiological range of temperatures between TRPM8 and TRPV1. PMID- 12077605 TI - Ubiquitination of histone H2B regulates H3 methylation and gene silencing in yeast. AB - In eukaryotes, the DNA of the genome is packaged with histone proteins to form nucleosomal filaments, which are, in turn, folded into a series of less well understood chromatin structures. Post-translational modifications of histone tail domains modulate chromatin structure and gene expression. Of these, histone ubiquitination is poorly understood. Here we show that the ubiquitin-conjugating enzyme Rad6 (Ubc2) mediates methylation of histone H3 at lysine 4 (Lys 4) through ubiquitination of H2B at Lys 123 in yeast (Saccharomyces cerevisiae). Moreover, H3 (Lys 4) methylation is abolished in the H2B-K123R mutant, whereas H3-K4R retains H2B (Lys 123) ubiquitination. These data indicate a unidirectional regulatory pathway in which ubiquitination of H2B (Lys 123) is a prerequisite for H3 (Lys 4) methylation. We also show that an H2B-K123R mutation perturbs silencing at the telomere, providing functional links between Rad6-mediated H2B (Lys 123) ubiquitination, Set1-mediated H3 (Lys 4) methylation, and transcriptional silencing. Thus, these data reveal a pathway leading to gene regulation through concerted histone modifications on distinct histone tails. We refer to this as 'trans-tail' regulation of histone modification, a stated prediction of the histone code hypothesis. PMID- 12077606 TI - TRPV3 is a temperature-sensitive vanilloid receptor-like protein. AB - Vanilloid receptor-1 (VR1, also known as TRPV1) is a thermosensitive, nonselective cation channel that is expressed by capsaicin-sensitive sensory afferents and is activated by noxious heat, acidic pH and the alkaloid irritant capsaicin. Although VR1 gene disruption results in a loss of capsaicin responses, it has minimal effects on thermal nociception. This and other experiments--such as those showing the existence of capsaicin-insensitive heat sensors in sensory neurons--suggest the existence of thermosensitive receptors distinct from VR1. Here we identify a member of the vanilloid receptor/TRP gene family, vanilloid receptor-like protein 3 (VRL3, also known as TRPV3), which is heat-sensitive but capsaicin-insensitive. VRL3 is coded for by a 2,370-base-pair open reading frame, transcribed from a gene adjacent to VR1, and is structurally homologous to VR1. VRL3 responds to noxious heat with a threshold of about 39 degrees C and is co expressed in dorsal root ganglion neurons with VR1. Furthermore, when heterologously expressed, VRL3 is able to associate with VR1 and may modulate its responses. Hence, not only is VRL3 a thermosensitive ion channel but it may represent an additional vanilloid receptor subunit involved in the formation of heteromeric vanilloid receptor channels. PMID- 12077607 TI - Dopamine neurons derived from embryonic stem cells function in an animal model of Parkinson's disease. AB - Parkinson's disease is a widespread condition caused by the loss of midbrain neurons that synthesize the neurotransmitter dopamine. Cells derived from the fetal midbrain can modify the course of the disease, but they are an inadequate source of dopamine-synthesizing neurons because their ability to generate these neurons is unstable. In contrast, embryonic stem (ES) cells proliferate extensively and can generate dopamine neurons. If ES cells are to become the basis for cell therapies, we must develop methods of enriching for the cell of interest and demonstrate that these cells show functions that will assist in treating the disease. Here we show that a highly enriched population of midbrain neural stem cells can be derived from mouse ES cells. The dopamine neurons generated by these stem cells show electrophysiological and behavioural properties expected of neurons from the midbrain. Our results encourage the use of ES cells in cell-replacement therapy for Parkinson's disease. PMID- 12077609 TI - Tobacco use in Tunisia: behaviour and awareness. AB - OBJECTIVE: To assess tobacco use and the awareness of and attitudes towards tobacco and its control in the adult population of Tunisia. METHODS: A cross sectional study was conducted in 1996 of a representative national sample of 5696 subjects aged 25 and over. Data were collected by means of a questionnaire in Arabic. FINDINGS: Tobacco use was reported by 30.4% of the respondents, of whom 24.6% smoked cigarettes and 5.8% consumed traditional tobacco, i.e. snuff, chewing tobacco and/or water pipe tobacco. Whereas 55.6% of men used tobacco, only 5.2% of women did so. Among men the proportion of tobacco users diminished with age as the rate of cessation increased. Among women, smoking peaked in the 35-54 age group. The proportion of men consuming traditional tobacco alone increased from 2.4% in the 25-34 age group to 20.4% in the 55+ age group; the corresponding values for women were 0.1% and 14.3%. Tobacco use was more widespread in rural than in urban areas and was relatively high among poorly educated men from economically deprived backgrounds. The use of tobacco was believed to be harmful to health by 98.6% of the respondents. Over 90% of the interviewees were aware that tobacco played a part in the development of heart disease. However, there were some gaps in awareness. A fear of cancer was expressed by 85% of the respondents, whereas only 5.6% were fearful of accidents. CONCLUSIONS: Informational and educational campaigns relating to tobacco control should be directed at individuals and communities, taking into account the gaps in awareness of the effects of tobacco on health. PMID- 12077608 TI - Early detection and response to meningococcal disease epidemics in sub-Saharan Africa: appraisal of the WHO strategy. AB - OBJECTIVE: To assess the sensitivity, specificity and predictive value positive of the WHO threshold strategy for detecting meningococcal disease epidemics in sub-Saharan Africa and to estimate the impact of the strategy on an epidemic at district level. METHODS: Data on meningitis cases at the district level were collected weekly from health ministries, WHO country and regional offices, and nongovernmental organizations in countries where there were epidemics of meningococcal disease in 1997. An epidemic was defined as a cumulative district attack rate of at least 100 cases per 100,000 population from January to May, the period of epidemic risk. The sensitivity, specificity and predictive value positive of the WHO threshold rate were calculated, and curves of sensitivity against (1 - specificity) were compared with alternatively defined threshold rates and epidemic sizes. The impact of the WHO strategy on a district epidemic was estimated by comparing the numbers of epidemic cases with cases estimated to have been prevented by vaccination. FINDINGS: An analysis was made of 48 198 cases reported in 174 districts in Benin, Burkina Faso, the Gambia, Ghana, Mali, Niger, and Togo. These cases were 80.3% of those reported from Africa to WHO during the 1997 epidemic period. District populations ranged from 10,298 to 573,908. The threshold rate was crossed during two consecutive weeks in 69 districts (39.7%) and there were epidemics in 66 districts (37.9%). Overall, the sensitivity of the threshold rate for predicting epidemics was 97%, the specificity was 95%, and the predictive value positive was 93%. Taken together, these values were equivalent or better than the sensitivity, specificity and predictive value positive of alternatively defined threshold rates and epidemics, and remained high regardless of district size. The estimated number of potential epidemic cases decreased by nearly 60% in the age group targeted for vaccination in one district where the guidelines were followed in a timely manner. CONCLUSION: The use of the WHO strategy was sensitive and specific for the early detection of meningococcal disease epidemics in countries of sub-Saharan Africa during 1997 and had a substantial impact on a district epidemic. Nevertheless, the burden of meningococcal disease in these countries remains formidable and additional control measures are needed. PMID- 12077610 TI - Using mortuary statistics in the development of an injury surveillance system in Ghana. AB - OBJECTIVE: To develop, in a mortuary setting, a pilot programme for improving the accuracy of records of deaths caused by injury. METHODS: The recording of injury related deaths was upgraded at the mortuary of the Komfo Anokye Teaching Hospital, Kumasi, Ghana, in 1996 through the creation of a prospectively gathered database. FINDINGS: There was an increase in the number of deaths reported annually as attributable to injury from 72 before 1995 to 633 in 1996-99. Injuries accounted for 8.6% of all deaths recorded in the mortuary and for 12% of deaths in the age range 15-59 years; 80% of deaths caused by injury occurred outside the hospital and thus would not have been indicated in hospital statistics; 88% of injury-related deaths were associated with transport, and 50% of these involved injuries to pedestrians. CONCLUSIONS: Injury was a significant cause of mortality in this urban African setting, especially among adults of working age. The reporting of injury-related deaths in a mortuary was made more complete and accurate by means of simple inexpensive methods. This source of data could make a significant contribution to an injury surveillance system, along with hospital records and police accident reports. PMID- 12077611 TI - Ectopic pregnancy in Conakry, Guinea. AB - OBJECTIVE: To assess the incidence of ectopic pregnancy (EP) in hospitals in Conakry, the capital of Guinea, West Africa. Data on EP incidence in developing countries are rare and often out of date, particularly in Africa. METHODS: A retrospective study was carried out, examining all cases of EP registered in the medical files of two referral maternity units at the Donka and Ignace Deen university hospitals between 1995 and 1999. FINDINGS: The EP incidence at the two maternity units increased from 0.41% to 1.5% of annual deliveries over this period. Haemoperitoneum was observed in most women, with tubal rupture in 93%; only 6 women received conservative treatment. CONCLUSION: The results suggest that the hospital-based incidence of EP per delivery has increased over the last decade in this West African capital, and that health professionals and public health officials in developing countries, especially those in Africa, should consider EP as a major obstetric problem for maternal morbidity. PMID- 12077612 TI - Education and acute coronary syndromes: results from the CARDIO2000 epidemiological study. AB - OBJECTIVE: As a measure of socioeconomic status, low educational level is positively associated with the risk of developing adverse health events. The aim of this study was to investigate the relationship between the level of education and the risk of developing non- fatal acute coronary syndromes. METHODS: During 2000 and 2001, 1619 randomly selected subjects from several regions of Greece were entered into a case-control study. Of these, 750 were patients with their first acute coronary heart syndrome event, and 869 were hospitalized controls with no cardiovascular disease in their medical history. Trends in cardiovascular risk factors were then examined across patient and control educational level by years of schooling. FINDINGS: In both patients and controls, education status was related to economic and occupation status, smoking habits, physical inactivity, alcohol consumption and non-compliance to treatment. After adjusting for these and other conventional risk factors, as well as for the effects of age and sex, we found that coronary risk increases by 82% (odds ratio (OR) = 1.82, P <0.05) for individuals with a lower level of education, and by 65% (OR = 1.65, P <0.05) for individuals with an average education, compared to those with an academic education. CONCLUSIONS: Although the least-educated subjects adopted a more adverse lifestyle than the more-educated subjects, the inverse association between education and coronary risk was independent from such factors. The inverse association may be due to psychosocial differences, and prospective cohort studies are needed to confirm or refute these results. PMID- 12077613 TI - The treatment gap and primary health care for people with epilepsy in rural Gambia. AB - OBJECTIVE: To study primary-level management for people with epilepsy in rural Gambia by means of community surveys. METHODS: After population screening was carried out, visits were made by a physician who described the epidemiology of epilepsy and its management. Gaps between required management and applied management were investigated by conducting interviews and discussions with people with epilepsy and their communities. FINDINGS: The lifetime prevalence of epilepsy was 4.9/1000 and the continuous treatment rate was less than 10%. The choice of treatment was shaped by beliefs in an external spiritual cause of epilepsy and was commonly expected to be curative but not preventive. Treatment rarely led to the control of seizures, although when control was achieved, the level of community acceptance of people with epilepsy increased. Every person with epilepsy had sought traditional treatment. Of the 69 people with active epilepsy, 42 (61%) said they would like to receive preventive biomedical treatment if it were available in their local community. Key programme factors included the local provision of effective treatment and community information with, in parallel, clarification of the use of preventive treatment and genuine integration with current traditional sources of treatment and advice. CONCLUSION: Primary-level management of epilepsy could be integrated into a chronic disease programme covering hypertension, diabetes, asthma and mental health. Initial diagnosis and prescribing could take place away from the periphery but recurrent dispensing would be conducted locally. Probable epilepsy etiologies suggest that there is scope for primary prevention through the strengthening of maternal and child health services. PMID- 12077614 TI - Can ivermectin mass treatments eliminate onchocerciasis in Africa? AB - OBJECTIVE: To elucidate the conditions in which mass treatment with ivermectin reduces the transmission of Onchocerca volvulus sufficiently to eliminate infection from an African community. METHODS: ONCHOSIM, a microsimulation model for onchocerciasis transmission, was used to explore the implications of different treatment intervals, coverage levels and precontrol endemicities for the likelihood of elimination. FINDINGS: Simulations suggested that control strategies based exclusively on ivermectin mass treatments could eliminate onchocerciasis. The duration of treatment required to eliminate infection depended heavily on the treatment programme and precontrol endemicity. In areas with medium to high levels of infection, annual mass treatments with 65% coverage for at least 25 years were necessary. Model predictions suggested that durations exceeding 35 years would be required if there were much heterogeneity in exposure to vector bites and, consequently, wide individual variation in microfilaria counts. If the treatment interval were reduced from 12 to 6 months the time for completion of the programme could be more than halved and elimination could be accomplished in areas of hyperendemicity, provided that the effects of each treatment would be the same as with annual treatments. However, it was doubtful whether high coverage levels could be sustained long enough to achieve worldwide eradication. CONCLUSION: Elimination of onchocerciasis from most endemic foci in Africa appears to be possible. However, the requirements in terms of duration, coverage, and frequency of treatment may be prohibitive in highly endemic areas. PMID- 12077615 TI - Trends and spatial distribution of deaths of children aged 12-60 months in Sao Paulo, Brazil, 1980-98. AB - OBJECTIVE: To describe trends in the mortality of children aged 12-60 months and to perform spatial data analysis of its distribution at the inner city district level in Sao Paulo from 1980 to 1998. METHODS: Official mortality data were analysed in relation to the underlying causes of death. The population of children aged 12-60 months, disaggregated by sex and age, was estimated for each year. Educational levels, income, employment status, and other socioeconomic indices were also assessed. Statistical Package for Social Sciences software was used for the statistical processing of time series. The Cochrane-Orcutt procedure of generalized least squares regression analysis was used to estimate the regression parameters with control of first-order autocorrelation. Spatial data analysis employed the discrimination of death rates and socioeconomic indices at the inner city district level. For classifying area-level death rates the method of K-means cluster analysis was used. Spatial correlation between variables was analysed by the simultaneous autoregressive regression method. FINDINGS: There was a steady decline in death rates during the 1980s at an average rate of 3.08% per year, followed by a levelling off. Infectious diseases remained the major cause of mortality, accounting for 43.1% of deaths during the last three years of the study. Injuries accounted for 16.5% of deaths. Mortality rates at the area level clearly demonstrated inequity in the city's health profile: there was an increasing difference between the rich and the underprivileged social strata in this respect. CONCLUSION: The overall mortality rate among children aged 12-60 months dropped by almost 30% during the study period. Most of the decline happened during the 1980s. Many people still live in a state of deprivation in underserved areas. Time-series and spatial data analysis provided indications of potential value in the planning of social policies promoting well-being, through the identification of factors affecting child survival and the regions with the worst health profiles, to which programmes and resources should be preferentially directed. PMID- 12077616 TI - Evaluation of two years of mass chemotherapy against ascariasis in Hamadan, Islamic Republic of Iran. AB - OBJECTIVE: To evaluate the mass treatment of ascariasis in rural areas of Hamadan Province, Islamic Republic of Iran. METHODS: A control programme in rural areas of Hamadan Province, which began in November 1997, involved giving all persons a single dose of 400 mg albendazole at intervals of three months. The efficacy of the treatment was evaluated by the formalin-ether concentration technique for stool examination and by the Stoll quantitative method. FINDINGS: The average rate of infection with Ascaris before treatment was 53.3%, ranging from 40% in Hamadan district to 75% in Toysercan. Two areas, Malayer and Nahavand, were excluded from the programme because the infection rates were only 13% and 4%, respectively. After two years of mass treatment the infection rate had decreased to 6%. The proportion of positive cases excreting only unfertilized eggs increased to 32%. No side-effects of mass treatment were observed. CONCLUSION: Systematic mass treatment giving high coverage proved to be very effective in the control of ascariasis, notwithstanding a lack of other preventive measures. PMID- 12077617 TI - Air travel and venous thromboembolism. AB - There has recently been increased publicity on the risk of venous thrombosis after long-haul flights. This paper reviews the evidence base related to the association between air travel and venous thromboembolism. The evidence consists only of case reports, clinical case-control studies and observational studies involving the use of intermediate end-points, or expert opinion. Some studies have suggested that there is no clear association, whereas others have indicated a strong relationship. On the whole it appears that there is probably a link between air travel and venous thrombosis. However, the link is likely to be weak, mainly affecting passengers with additional risk factors for venous thromboembolism. The available evidence is not adequate to allow quantification of the risk. There are insufficient scientific data on which to base specific recommendations for prevention, other than that leg exercise should be taken during travel. Further studies are urgently needed in order to identify prospectively the incidence of the condition and those at risk. PMID- 12077618 TI - Definition and measurement of reproductive health. AB - An internationally agreed conceptual definition of reproductive health is applied to the development and testing of practical indicators for use in the community. Basic criteria are proposed for an interview-based tool to measure reproductive health -- as opposed to morbidity or mortality -- adapting methods from the health status measurement field. Proposed domains and indicators linked to the definition of reproductive health adopted at the International Conference on Population and Development (ICPD) should be comparable across and within diverse populations. Two sets of domains that describe reproductive health are recommended for further development and testing, seven domains that focus directly on health and six others that assess related areas of well-being. PMID- 12077619 TI - Eliminating iodine deficiency disorders--the role of the International Council in the global partnership. AB - Iodine deficiency is the most common preventable cause of brain damage. WHO estimates that some 2.2 billion people are at risk from iodine deficiency in 130 countries. A programme of universal salt iodization was established in 1994 with the aim of eliminating the problem by 2000. This paper reports progress in this field, with particular reference to the primarily scientific role of the International Council for Control of Iodine Deficiency Disorders, a nongovernmental organization founded in 1986. It is now a multidisciplinary network of 600 professionals in 100 countries. PMID- 12077622 TI - Diabetic retinopathy: time for action. No complacency please! PMID- 12077624 TI - Global response to antimicrobial resistance. PMID- 12077623 TI - Societal variables are central to effective HIV intervention models. PMID- 12077625 TI - Crisis in East European health systems--"Europe's best kept secret". PMID- 12077626 TI - HIV does cause AIDS but it's hard to prescribe the drugs, says South Africa's ANC. PMID- 12077627 TI - AIDS growth in India can be stopped in five years, claims government. PMID- 12077628 TI - Nepal deworming programme ready to go worldwide. PMID- 12077631 TI - Access to HIV drugs: reliable products listed. PMID- 12077633 TI - Comparison between midline incision and limited right skin crease incision for right-sided colonic cancers. AB - We compared the postoperative recovery parameters between patients undergoing curative surgery for right-sided colonic carcinoma using a limited skin crease incision and a traditional midline incision. A retrospective study was carried out analyzing clinical records and histopathological reports for all patients operated in one colorectal surgical unit for cancer of the right colon over a 2 year period. Palliative procedures were excluded. We analyzed demographic details, operative data (length of incision and time of operation), recovery parameters (time to parenteral analgesia, time to first oral fluid intake, time to first solid meal, time to discharge) and oncological parameters (lymph node harvest and resection margins). A total of 123 patients were analyzed, 61 with a midline incision and 62 with a skin crease incision. Demographic and tumour data (number of lymph nodes resected and resection margins) as well as postoperative complications were similar between the two groups. Wound length was significantly longer in the midline incision group (median, 20 cm vs. 10 cm; p<0.0005), as was the duration of surgery (median, 60 min vs. 45 min; p0.0005). With regard to postoperative recovery, the skin crease incision group had a significantly quicker return of bowel function ( p<0.0005), shorter time to oral fluid ( p<0.001) and solid food ( p<0.0005) intake, and shorter hospital stay ( p<0.0005) than the midline incision group. There was no statistically significant difference between the two groups concerning postoperative narcotic requirements. In conclusion, the limited skin crease approach for right colon cancer resection is technically feasible and safe. It can achieve the same standards of tumour resection and clearance as the vertical midline approach while reducing postoperative recovery. PMID- 12077634 TI - Experience in management of Fournier's gangrene. AB - We report our experience and results in the management of Fournier's gangrene. Fournier's gangrene is a synergistic infective necrotizing fasciitis, which involves perianal, perineal and genital regions, originated mostly from colorectal and genitourinary sources. Charts and records from 28 patients with Fournier's gangrene diagnosed between 1993 and 1997 were reviewed. The mean patients age was 57.8 years (range, 22-82 years); mean hospital stay was 19 days. Eighteen patients (64.3%) were diabetic. The most common source of gangrene was ischiorectal abscess in 22 patients (78.6%). Colostomy was performed on 14 patients (50%) and cystostomy on 7 patients (25%). Ten patients (35.7%) died because of sepsis. In conclusion, medical and surgical treatment should be aggressive. Colostomy should only be performed if sphincter complex is damaged. Multidisciplinary management is mandatory, because of high morbidity and mortality. PMID- 12077635 TI - Effect of oral diazepam on anal continence after low anterior resection: a preliminary study. AB - The aim of the present study was to investigate the effect of oral diazepam on anal incontinence after low anterior resection for rectal cancer. Five patients with persistent incontinence after low anterior resection for rectal cancer (median level of anastomosis was 4.0 cm from the anal verge) were treated with oral diazepam (2 mg/day) 9-90 months after surgery. Grade and frequency of anal incontinence, the need for a protective pad and changes of lifestyle were recorded to the Cleveland Clinic's continence grading scale; anorectal manometry was performed before and after 3 months of treatment. All patients improved on treatment although occasional minor soiling persisted in two patients. Continence score improved from 14 (median, range 9-16) to 0 (range 0-12) after taking diazepam. Improvement occurred within a week after administration of diazepam. Although the patients improved symptomatically, anorectal manometry failed to demonstrate any significant changes. In conclusion, oral administration of diazepam may be worthwhile in the attempt to improve anal continence after low anterior resection. PMID- 12077636 TI - Management of hemorrhoidal disease in patients with chronic spinal cord injury. AB - Hemorrhoidal disease is a common pathology in patients with chronic spinal cord injury (SCI). We describe our experience with the primary approach to this problem at the Proctology Division of the Sheba Medical Center. We treated 29 patients (26 men) with paraplegia due to SCI between 1995 and 1999. The mean age was 49 years (range, 22-74 years). All patients had hemorrhoids in stages ranging between II and IV. Main complaints were rectal bleeding (83%), difficulties in evacuation (38%) and discomfort or pain (28%). Eleven patients (38%) were treated conservatively (e. g. diet, hygiene and laxatives), while 18 patients (62%) underwent either banding or sclerotherapy of hemorrhoids or both. No major complication were observed. In 28 of 29 patients (96%), there was a significant reduction or cessation of bleeding and/or relief of symptoms; one patient (3%) required hemorrhoidectomy. Of the 28 successful treatments, 16 (57%) had partial reduction of bleeding or relief of symptoms, while in 12 (43%) response was complete. Of those who were treated conservatively, 9 (82%) had partial and 2 (18%) had complete relief of symptoms. Of those who had banding/sclerotherapy, 7 (41%) had partial and 10 (59%) had complete relief. We also examined the effect of perianal sensation on the treatment outcome. Of 16 patients with complete anesthesia, 11 (69%) had partial and 5 (31%) had complete relief, whereas of the 12 patients with preserved sensation, 5 (42%) had partial and 7 (58%) had complete relief. In conclusion, the approach of banding or sclerotherapy of hemorrhoids in SCI patients is safe and effective. When sensation of the perianal region is preserved, the outcome seems to be better. The cause of SCI has no impact on the treatment results. There was no difference in the outcome of treatment between patients with stage II and stage III hemorrhoids; patients with stage IV hemorrhoids seem to do worse than those with stages II and III. PMID- 12077637 TI - Subtotal colectomy with antiperistaltic cecorectal anastomosis. AB - We assessed the functional results achieved with an antiperistaltic end-to-end cecorectal anastomosis (CRA) after subtotal colectomy. A total of 34 patients with colonic cancer, inflammatory bowel disease or chronic constipation were treated with subtotal colectomy and cecorectal anastomosis. The postoperative mortality was nil; no major postoperative complication was registered. At a mean 60-month follow-up (range, 12-92), 32 patients were alive, showed no diarrhea nor incontinence, were completely relieved from previous symptoms and abdominal discomfort, and had a normal diet. In conclusion, subtotal colectomy with end-to end antiperistaltic CRA seems to have a role in selected cases for the treatment of inflammatory bowel diseases, colon tumors and slow-transit constipation. PMID- 12077638 TI - Sacrococcygeal pilonidal sinus disease with Limberg flap repair. AB - Sacrococcygeal pilonidal sinus disease affects younger persons, resulting in long term loss of productive power. It also has a high rate of morbidity because there is no standard treatment approved by all surgeons. In this study, early and late results of 85 cases treated with excision and Limberg flap in our clinic were investigated. The patients were followed for 9-120 months. The complication rate was 4.7% and the long-term relapse rate was 3.5: Insufficient personal hygiene and inadequate excision were the main causes for recurrence. On the basis of the literature and our previous experiences, we conclude that the Limberg flap is the ideal treatment for sacrococcygeal pilonidal sinus disease with low morbidity, mortality and recurrence rates. PMID- 12077639 TI - Minimally invasive transanal surgery for localized rectal carcinoid tumors. AB - Local excision is often fully justified for rectal carcinoid tumors. However insufficient surgical field and difficult access to proximal tumors have been drawbacks in performing pre-existing local excision procedures. A novel local excisional technique called minimally invasive transanal surgery (MITAS) has been experimented for local removal of carcinoid tumors in the rectum. A specially designed anal retractor connected to the Octopus retractor holder was used and an ENDO-stapler allowed the simultaneous excision and anastomosis to be performed. Eight patients with carcinoid tumors in the rectum (4 tumors in the upper rectum) underwent MITAS. Median distance from anal verge to proximal tumor was 6.5 cm (range, 5-12 cm). The median diameter of the tumor was 9 mm. Median operative time was 18.5 minutes and blood loss was minimal. No analgesics were needed postoperatively, and there was no morbidity or mortality. Full-thickness excision of the rectum was accomplished and the tumors confined in the submucosa were demonstrated histologically to be with free surgical margins. No recurrences have been observed with a median follow-up period of 39 months. The technique facilitates total excisional biopsy for rectal carcinoid tumors and reduces operative time, blood loss and complications. PMID- 12077640 TI - The outcome of transanal advancement flap repair of rectovaginal fistulas is not improved by an additional labial fat flap transposition. AB - Transanal advancement flap repair (TAFR) has been advocated as the treatment of choice for patients with low rectovaginal fistulas. Recently, several studies have reported a significantly lower healing rate. We also encountered low healing rates after TAFR. In an attempt to improve our results, we added labial fat flap transposition (LFFT) to the TAFR of rectovaginal fistulas. The aim of the present study was to evaluate the outcome after TAFR and to investigate the impact of an additional LFFT. Between 1991 and 1997, 21 consecutive patients of median age 33 years underwent TAFR. The etiology of the fistulas was: obstetric injury (n=9), cryptoglandular abscess (n=8) and wound infection after anterior anal repair (n=4). The first 9 patients underwent TAFT with (n=3) or without (n=6) anterior anal repair. In the following 12 patients, LFFT was added to the advancement flap. In 4 of these a concomitant anterior anal repair was performed. The median follow-up was 15 months. The overall healing rate was 48%. In the first 9 patients, in whom no additional LFFT was performed, the rectovaginal fistula healed in 4 cases (44%). In the following 12 patients in whom an additional LFFT was performed, a similar healing rate was observed (50%). In conclusion, the outcome of transanal advancement flap repair of rectovaginal fistulas is poor. Addition of a labial fat flap transposition does not improve this outcome. PMID- 12077641 TI - The management of presacral masses in the adult. AB - Presacral (retrorectal) tumors are particularly rare in the adult. There is difficulty in the performance of diagnostic biopsy, and specialized imaging is required to plan surgical extirpation. This review assesses their incidence and classification as well as the principles involved in their diagnosis and surgical management. PMID- 12077642 TI - Cauterization-plication operation in the treatment of complete rectal prolapse. AB - The current communication presents a simple technique for treatment of complete rectal prolapse (CRP). The study included 28 patients presenting with CRP (mean age, 36.4 years; 4 children 2-12 years; 17 female and 11 males). Fourteen patients had fecal incontinence. With the patient under general anesthesia in lithotomy position, the prolapsed rectum was pulled outside the anal canal, the mucosa was cauterized in vertical lines and the exposed muscle layer was plicated by 2/0 coated Vicryl sutures. Posterior levatorplasty was done in 14 adult patients in whom the length of prolapsed segment was more than 10 cm and who were incontinent due to a wide levator hiatus. The postoperative follow up was 31.6+/ 14.8 months (mean+/-SD). Five had postoperative mucosal prolapse and one had recurrence 3 months of operation. Mucosal plication was performed for the five patients and the operation was redone for the recurrent patient. Fecal impaction, stricture and fistula formation were not encountered. The technique is simple, easy and with minimal complications. PMID- 12077643 TI - Duplication of appendix vermiformis: a case in a child. AB - Appendiceal anomalies are extremely rare malformations that are usually found in adult population as an incidental finding during laparotomy due to another reason. When appendiceal duplications are detected in childhood, almost all the patients have serious associated intestinal, genito-urinary or vertebral malformations. Presented herein are a case of a huge (15 x 13 cm mass) appendiceal duplication in a child causing acute abdomen without an accompanying pathology, its differentiation from appendiceal and caecal diverticulum, and classification of appendiceal duplications with the review of literature. PMID- 12077644 TI - Dynamic imaging of pelvic floor with transperineal sonography. PMID- 12077645 TI - Risks, costs, and compliance limit colorectal adenoma surveillance: lessons from a randomised trial. PMID- 12077648 TI - Combined seton-double flap procedure for complex high anal fistula. PMID- 12077649 TI - Current orthopedic problems in Korea. PMID- 12077650 TI - An interdisciplinary and comprehensive approach for treatment of musculoskeletal tumors in the bone and joint decade. PMID- 12077651 TI - Prevalence of abnormal findings in magnetic resonance images of asymptomatic knees. AB - One hundred fifteen asymptomatic Japanese volunteers aged from 13 to 76 years were examined by magnetic resonance imaging (MRI) to evaluate age-related meniscal degeneration and to determine the prevalence of discoid menisci. Each meniscus was graded in the anterior and posterior portions according to intrameniscal MRI signals. Discoid meniscus was diagnosed if a bow-tie configuration was noted on three or more contiguous sagittal sections. Meniscal abnormalities on MRI became more prevalent with age in both men and women. The posterior horn of the medial meniscus showed a significantly higher prevalence of degeneration than other parts of the meniscus. Discoid menisci were noted in 15 subjects (13% prevalence), always representing the lateral meniscus. Subchondral changes were observed in 13 subjects more than 40 years old, mostly women, and were located in the medial compartment. These abnormalities were not correlated with severity of degeneration in the posterior portion of the medial meniscus. This study demonstrates considerable prevalence of meniscal abnormalities in asymptomatic Japanese subjects. PMID- 12077652 TI - Estimation of forces at the interface between an artificial limb and an implant directly fixed into the femur in above-knee amputees. AB - This article describes the method used for estimating the forces and moments, acting during locomotion, at the interface between an artificial leg and an implant directly fixed into the femur, in above-knee amputees. Twelve transfemoral amputees completed a predefined gait assessment during which kinetic (ground reaction loads and torque) and kinematic (limb orientation) gait data were recorded. A developed mathematical model enabled the ground reaction forces to be translated to the level of amputation. It is assumed that the loads calculated at the stump-socket interface would approximate those experienced by the proposed implant. The longitudinal force and the moment in the sagittal plane were the two most significant loads at the stump. These data were essential to obtain to facilitate the analysis of stress arising at the implant-bone interface that has been subsequently undertaken. PMID- 12077653 TI - The relationship between distraction length and treatment indices during distraction osteogenesis. AB - To evaluate the indices used to compare the results of limb lengthening with distraction osteogenesis, we analyzed the relationship between the lengthening and external fixation period in a series of 28 patients (30 limbs). The average lengthening was 4.6 cm (range, 2.0-8.9). The average age at operation was 16.6 years (range, 4-47). Limbs treated included 15 femurs and 15 tibias of 19 male and 9 female patients. The external fixation period and the maturation period had a direct linear relationship with length gain. There was a negative hyperbolic relationship between the external fixation index, maturation index, and length gain. Segments that were lengthened 0-3.0 cm had a maturation index of 38.3 days/cm and external fixation index of 57.6 days/cm, both of which were significantly higher than for those lengthened more than 3.0 cm. When comparing the maturation and external fixation indices between different patients or studies, the length of the distraction must be taken into consideration. PMID- 12077654 TI - Clinical significance of thallium-201 scintigraphy in bone and soft tissue tumors. AB - We evaluated sequential thallium scans on both early images (EI) and delayed images (DI) for 62 patients who had bone and soft tissue lesions. The purpose was to determine whether this technique could be used to ascertain accurately whether lesions were malignant or benign and to predict the response to chemotherapy. The thallium-201 chloride (201Tl) accumulation in malignant tumors and benign lesions was statistically different. Sensitivity, specificity, and accuracy for 201Tl scans in detecting malignant tumors was 94%, 65%, and 82%, respectively, for EI, and 94%, 85%, and 90%, respectively, for DI. On multivariate analysis, significant independent factors for 201Tl uptake were malignant lesions on EI and DI and high cellularity on EI. Thirteen patients with malignant bone and soft tissue tumors underwent 201Tl scans before and after preoperative chemotherapy. There was a good correlation between percentage of tumor necrosis and percentage change of accumulation in lesion-to-normal tissue ration, and the correlation coefficient was higher on EI ( r = 0.801) than on DI ( r = 0.664). These results support the notion that 201Tl scintigraphy, although showing some false-positive and false-negative findings, is a useful tool in the evaluation of either malignant tumors or benign lesions. Furthermore, 201Tl scans on EI provide benefit concerning the evaluation of chemotherapeutic response in patients with malignant bone and soft tissue tumors. PMID- 12077655 TI - A modified technique for anterior multilevel cervical fusion. AB - Anterior cervical fusion with interbody bone graft and anterior plating is commonly performed. Unfortunately, the plate has been reported to shield the graft from loading, thus reducing fusion rates. Interbody fusion cages have been effective in the lumbar spine and have gained acceptance in the cervical spine. Twenty-five patients underwent anterior cervical fusion with this modified technique. All patients received anterior diskectomy and corpectomy, placement of an interbody fusion cage packed with corpectomy bone, and application of an anterior cervical plate. Fusion was defined by radiographic evidence of trabecular bone bridging through the cage. No external bracing was used except soft collars as needed. Pre- and postoperative pain scales were completed and statistically analyzed using paired t tests. There were no cases of pseudoarthrosis or major neurological, vascular, or wound complications. There was one case of mild dysphagia that remained unresolved. Mean operative time was comparable to standard instrumented multilevel cervical fusion surgeries. Visual analogue pain scales were significantly improved following surgery. The advantages of using interbody cages with anterior plating include immediate stability and support, elimination of donor site pain from iliac crest bone autograft, and a decrease in pseudoarthrosis by halving the number of fusion surfaces. PMID- 12077656 TI - Clinical results of posterolateral fusion for degenerative lumbar spinal diseases: a follow-up study of more than 10 years. AB - The objective of this retrospective study was to evaluate the long-term clinical outcomes and the effects on unfused motion segments of posterolateral fusion. This study involved 35 cases (37 intervertebral levels) of posterolateral fusion performed to treat degenerative lumbar spinal diseases. There were 20 male and 15 female patients ranging in age from 30 to 67 years, with a mean age of 49 years. The postoperative period ranged from 10 years to 17 years and 8 months, with a mean period of 13 years. The clinical outcomes were evaluated using the Japanese Orthopaedic Association (JOA) score for assessment of treatment for low back pain. The effects on unfused motion segments were investigated with radiographic and magnetic resonance (MR) imaging. Postoperative satisfactory improvement (mean recovery rate, 66.9%) reached a plateau at 1 year and was maintained at final follow-up. Radiographically, the union rate was 86.5%. There were few cases of induced instability of unfused motion segments. On MR imaging, increased signal intensity in both T(1)- and T(2)-weighted images was seen in the paravertebral muscles in 15 of 20 cases (75.0%). Posterolateral fusion is a useful technique for the treatment of degenerative lumbar spinal diseases. Clinical outcomes were stable throughout follow-up. Instability of unfused motion segments rarely occurred. PMID- 12077657 TI - Bone mineral density after the removal of intramedullary nails: a cross-sectional and longitudinal study. AB - We measured bone mineral density (BMD) using dual X-ray absorptiometry (DXA) at several sites in both fractured and nonfractured limbs in eight patients with femoral shaft fracture and six with tibial shaft fracture at the time of the intramedullary (IM) nail removal. Seven patients were followed up for an average of 13 months. The BMD at the proximal part of the femur and tibia was from 3% to 11% lower in the fractured side as compared to the nonfractured side. The greatest bone loss (13%-21%) was found in the operated distal tibia of the patients with tibial shaft fractures. At the fracture site of the femur, BMD was 10.5% ( P < 0.05) higher, possibly owing to fracture callus formation, whereas tibial shaft BMD was not increased. However, a calculated apparent volumetric BMDvol at the fracture site was 15%-16% decreased. Although BMDs of the fractured side almost reached the baseline level of the nonfractured side (96.9%-102.1%) by the final follow-up (>12 months), the absolute deficit was still 3%-9%. Surprisingly, significant BMD increases (5%-6%) were also detected in all proximal femoral measurement sites of the contralateral limb, which indirectly suggests that the uninjured limb may also suffer from bone loss after lower extremity trauma. We conclude that clinically important bone loss exists in the proximal femur and proximal and distal tibia of the fractured limb at the time of IM nail removal. Although areal BMD was higher at the femoral fracture site, the lower apparent volumetric BMDvol suggests decreased mineralization and reduced strength of the fracture site. Although the present results do not suggest special recommendations for restricted weight bearing after the removal of IM nails, the relationship between decreased bone density and increased risk of fractures should be borne in mind. PMID- 12077658 TI - Bone mineralization gradient at the callotasis site. AB - Thirteen patients (18 lengthenings; mean age at operation, 11.0 years) underwent dual-energy X-ray absorptiometry (DEXA) scanning weekly during the distraction phase, at 2-week intervals until removal of the fixator, and at the time of each outpatient visit after removal of the apparatus for a median of 353 days. The three transverse regions remained significantly different in the 7 achondroplastic patients throughout the study, but the difference among these regions became nonsignificant by fixator removal in the 11 limb length discrepancy (LLD) patients. The most proximal region was significantly more mineralized throughout the study in the achondroplastic patients. The central region became the region of highest mineralization in the LLD patients by week 16 after removal of the fixator. The three longitudinal regions showed significantly different mineralization at all time points except at fixator removal. The central and medial regions always showed the highest mineralization. The mechanical characteristics of the fixator and the biomechanical features of the lengthening site may account for the mineralization gradient reported in this study and should probably be taken into account when planning removal of the fixator and subsequent weight-bearing. PMID- 12077660 TI - Coordination of the anterior and posterior cruciate ligaments in constraining the varus-valgus and internal-external rotatory instability of the knee. AB - Tension along both cruciate ligaments was measured simultaneously under various loading conditions, and the interaction of these ligaments as constraints on knee instability was analyzed. Six fresh cadaveric knees were used. The attachments for both cruciate ligaments were detached from the femur and reattached to their original positions using metal plates equipped with 12 strain gauges. Each knee was moved under various loading conditions, and changes in tension along the cruciate ligaments were recorded simultaneously using the output of the strain gauges. Under varus torque, tension along the anterior cruciate ligament increased near full extension whereas that along the posterior cruciate ligament increased near 90 degrees of flexion. Similar results were obtained under valgus torque. Under internal rotatory torque, a pattern similar to that under varus torque was also observed. Under external rotatory torque, no remarkable changes in tension were observed along either cruciate ligament. Thus, we conclude that both the anterior cruciate ligament and the posterior cruciate ligament cooperate to control varus-valgus and internal rotatory instabilities of the knee, and that the constraining function is transferred from the anterior cruciate ligament to the posterior cruciate ligament as the knee joint is flexed. PMID- 12077659 TI - Characteristics of anterior tibial translation with active and isokinetic knee extension exercise before and after ACL reconstruction. AB - The aim of this study was to investigate the biomechanical characteristics of anterior tibial translation (ATT) in anterior cruciate ligament (ACL)-deficient or -reconstructed knees with active and isokinetic knee extension exercise. Forty nine patients with unilateral isolated ACL-deficient knees were enrolled. Follow up examinations were carried out at a mean of 24 months postoperatively. An electrogoniometer system was applied to compare the amount of ATT in ACL deficient and -reconstructed knees. For both active and isokinetic knee extension, the mean ATT of ACL-deficient knees was considerably greater than that for the normal side, within a range of flexion 0 degrees -70 degrees and 0 degrees -60 degrees, respectively. In contrast, no mean ATT differences were seen during both active and isokinetic exercise from 90 degrees to 0 degrees at follow up. Within a range of flexion between 50 degrees and 70 degrees, the side-to-side difference in ATT with active knee extension was significantly greater than that with isokinetic extension in ACL-reconstructed knees. These results suggest that the amount of ATT is significantly improved with both active and isokinetic exercise, postoperatively. However, postoperative ATT with isokinetic extension is smaller than that with active knee extension from 50 degrees to 70 degrees. PMID- 12077661 TI - Effect of hyperthermia by magnetite cement on tumor-induced bone destruction. AB - Our study focuses on the antineoplasmic action of hyperthermia. In this study, use of a heat-generating cement was exploited for the management of pathological and impending fractures in malignancies. The cement contains magnetic powder in ceramic particles and generates well-regulated heat when a magnetic field is applied externally. Eighteen rabbits were inoculated with blocks of VX2 tumor into one of their tibia. One week after the procedure, 9 rabbits were exposed to a magnetic field (HT group) while the remaining 9 rabbits were not (non-HT group). In rabbits killed between 20 and 42 days after the VX2 inoculation, the circumference of the operated leg was 7.9 +/- 0.3 cm and 10.2 +/- 1.0 cm (mean +/ SE) in HT and non-HT groups, respectively ( P < 0.05). Histological findings revealed the regressive change in tumor tissue of the HT group. By radiographs, pathological fractures and cortical bone destruction were seen in 5 and 8 rabbits in the non-HT group, respectively, but in the HT group these effects were absent in all the rabbits except 1 in which a definite diagnosis has not been made. Our findings demonstrate that controlled hyperthermia therapy using a newly developed bone cement suppresses tumor growth and prevents local bone destruction caused by VX2 tumors. PMID- 12077662 TI - Effect of decreased physical activity on bone mass in exercise-trained young rats. AB - The aim of this study was to determine whether decreased physical activity in exercise-trained young rats would result in a lower rate of bone gain or a reversal of the benefits of exercise. Thirty-five female Wistar rats, 6 weeks of age, were randomized into seven groups: 7 weeks of exercise (7EX), 7 weeks of sedentary control (7CN), 11 weeks of exercise (11EX), 7 weeks of exercise followed by 4 weeks of exercise cessation (7EX4C), 7 weeks of exercise followed by 4 weeks of decreased exercise frequency (7EX4F), 7 weeks of exercise followed by 4 weeks of decreased exercise intensity (7EX4I), and 11 weeks of sedentary control (11CN). The running intensity (speed) and duration were 25 m/min for 60 min/day at a frequency of 5 days/week. During the last 4 weeks, exercise frequency was reduced to 1 day/week in the 11EX4F group, and exercise intensity (speed) was reduced to 12 m/min in the 7EX4I group. After each period of exercise, the bone mineral content (BMC) of the proximal, middle, and distal tibiae, determined by dual-energy X-ray absorptiometry (DXA), was significantly greater in the 7EX and 11EX groups than in the 7CN and 11CN groups, respectively, but it was significantly lower in the 7EX4C group than in the 11EX group and did not differ significantly from the values of the 11CN group. Although the BMC of the proximal and middle tibiae did not differ significantly among the 7EX4F, 7EX4I, 7EX4C, and 11CN groups, the BMC of the distal tibia was significantly greater in the 7EX4F and 7EX4I groups than in the 11CN group and tended to be greater than in the 7EX4C group. The results of this study suggest that the effect of decreased exercise intensity and frequency on bone mass appears to be site specific in the tibia of the exercise-trained young rats. This study shows that exercise-trained young rats lose the benefits gained from exercise when exercise is completely ceased, resulting in the reduction of bone mass to levels that do not differ significantly from those of sedentary controls. At least, continuous exercise appears to be necessary for the maintenance of high bone mass. PMID- 12077663 TI - Effects of vitamin K2, vitamin D, and calcium on the bone metabolism of rats in the growth phase. AB - Using 168 female Sprague-Dawley rats, we determined whether the peak bone mass could be increased, and which drugs would be effective in suppressing the rate of decrease in bone mass by continuous administration from childhood. At the age of 3 months, these 168 rats were divided into five groups depending on the type of diet that they were fed (control, regular; group A, vitamin K2; group B, vitamin D; group C, high calcium; group D, vitamins D and K2 and high calcium) and kept to the age of 16 months. Dual-energy X-ray absorptiometry (DXA) was used to measure the bone mineral density of the femoral epiphysis and microcomputed tomography (CT) to analyze its fine structure. The average bone mass increased rapidly with age and reached a peak at the age of 8 months. Peak bone mass for the four drug administration groups was higher than that for the control group. Among these four groups, the peak bone mass was the highest in group C and the rate of decrease the smallest in group D. The results of the present animal study suggest that the peak bone mass of humans can be raised by consuming sufficient amounts of vitamins K2 and D and calcium continuously from childhood, and that this diet will suppress the rate of decrease in bone mass, thus ultimately preventing bone fractures caused by osteoporosis. PMID- 12077664 TI - Mechanical properties of the femur after injection of calcium phosphate cement containing arbekacin sulfate and polylactic acid in a rat model of experimental osteoporosis. AB - We developed a calcium phosphate cement containing arbekacin sulfate and polylactic acid (CPC95AP) and examined its effects on bone strength by injecting the CPC95AP into the femoral medullary cavity in model rats with osteoporosis. Favorable cortical bone strength was maintained for more than 6 months when CPC95AP was injected into the femoral cavities. This phenomenon was more significant in the weakened femur of osteoporotic rats. The reasons for the increased bone strength are thought to be that the CPC95AP filling increased the geometric moment of inertia and that the binding of CPC95AP with the bones increased Young's modulus. The most important factor is believed to be the increased Young's modulus. The binding of CPC95AP with the bones after filling the femur cavities compensates for the weak extensional strength of CPC95AP, and provides an opportunity to utilize the compressive strength, which is the advantage of CPC95. CPC95AP has antibiotic properties and biocompatibility, and it increases the mechanical strength of the weakened bone; therefore, it is considered useful in bone filling and stiffening. PMID- 12077665 TI - Comparative effects of intermittent administration of human parathyroid hormone (1-34) on cancellous and cortical bone loss in tail-suspended and sciatic neurectomized young rats. AB - We compared the effects of intermittent administration of human parathyroid hormone (PTH) (1-34) on tibial cancellous and cortical bone loss in tail suspended and sciatic neurectomized young rats. Forty-eight 6-week-old male Wistar rats were randomly divided into six groups with eight animals each: age matched controls (AMC), tail suspension (TS), sciatic neurectomy (NX), AMC + PTH, TS + PTH, and NX + PTH. Fifteen days after the start of the experiment, the proximal tibia and tibial shaft were processed for cancellous and cortical bone histomorphometric analyses, respectively. The reduction of cancellous bone volume (BV/TV) was significantly greater in the TS group than in the NX group, whereas the reduction of percent cortical area (Ct Ar) did not differ significantly between the TS and NX groups. Administration of human PTH to rats in the TS and NX groups increased BV/TV to a level significantly higher than that of the AMC group. Administration of human PTH to rats in the NX group significantly increased percent Ct Ar, but percent Ct Ar of the NX group was still significantly lower than that of the AMC group. Administration of human PTH to rats in the TS group did not significantly affect percent Ct Ar. These findings suggest that intermittent administration of human PTH (1-34) at the dose we used may completely prevent cancellous bone loss both in TS and NX young rats, and that it may not affect cortical bone loss in TS young rats but only attenuate it in NX young rats. PMID- 12077666 TI - Low-velocity gunshot fractures of the tibia. AB - A gunshot-related fracture is a unique type of open fracture. Bullet-induced fractures of the tibia may be encountered in any urban or rural setting as a result of hunting or cleaning mishaps or, perhaps more frequently, of criminal activity. For surgeons with limited experience, there is a bewildering range of apparently contradictory advice concerning their management. This article reports a case of a 53-year-old man with a highly comminuted bullet-induced shaft fracture of the tibia received during hunting activity. The patient was treated with a unilateral external fixator and vascularized flap for skin reconstruction. Six months after surgery, the external fixator was removed at clinical and radiological evidence of healing. At 1 year follow-up, the patient obtained complete healing and returned to normal activity. PMID- 12077667 TI - Computer-assisted anterior spinal surgery for a case of recurrent giant cell tumor. AB - A computer-assisted image guidance system has recently been used for posterior spinal surgery. We applied this system to anterior revision surgery of the cervicothoracic junction for a patient with recurrent thoracic spinal giant cell tumor. Anterior computer-assisted spinal surgery was achieved by attaching reference markers to threaded pins inserted into a vertebral body. The locations of anatomic structures in the surgical field of this patient were difficult to identify because of previous surgery. Both accurate resection of the tumor and anterior fusion with iliac bone autograft between C6 and T3 were successfully performed using a computer-assisted image guidance system. This system is useful for anterior spinal surgery because it enables a surgeon to perform safe and accurate surgery. PMID- 12077668 TI - Decreased interposition after closed reduction in the hip joint: a case of developmental dislocation of the hip observed by MRI. AB - A 4-month-old Japanese girl with a developmental dislocation of the right hip was treated with manual reduction under general anesthesia followed by cast immobilization. Serial magnetic resonance imaging (MRI) revealed that a large interposition of fat tissue in the acetabulum, observed immediately after cast immobilization, became remarkably small in size as the congruity improved after 3 weeks. These MRI findings demonstrate the mechanism of a "squeezing phenomenon" after reduction of the hip. PMID- 12077669 TI - Sciatic nerve palsy after cementless total hip arthroplasty. Treatment by modular neck and calcar shortening: a case report. AB - This is a case report of sciatic nerve palsy after total hip arthroplasty. Although the patient's symptoms became worse postoperatively, full recovery occurred after shortening of the calcar and femoral neck length. For acute sciatic nerve palsy patients with worsening of symptoms in the postoperative course in spite of hip and knee flexion, reexposure for early recognition of the sciatic nerve condition and reoperation by shortening the femoral neck may be an option. PMID- 12077670 TI - Slipped capital femoral epiphysis following contralateral infantile Blount's disease. AB - Slipped capital femoral epiphysis (SCFE) and Blount's disease are reported to have a common etiology, but there is only one report describing two cases in which adolescent Blount's disease coexisted with SCFE. In this article, we describe a case of SCFE following contralateral infantile Blount's disease in an 11-year-old boy. This report is the first known case of SCFE associated with infantile Blount's disease. In this patient, pelvic tilt caused by leg length discrepancy associated with infantile Blount's disease and possible general weakness of the growth plate may be related to the occurrence of SCFE. PMID- 12077671 TI - Tackler's bony spur in sumo wrestlers: a report of two cases. AB - We examined two sumo wrestlers who had tackler's bony spur. In football players, these spurs are secondary to subperiosteal hematoma formation from repeated direct blows by an opponent or from ill-fitting shoulder pads. The initial charge of sumo wrestling is similar to a tackle in American football. Our two patients had had repeated direct blows by opponents on the initial wrestling charge. The tackler's bony spurs of our two patients were located on the lateral aspect of the humeral shaft just distal to the deltoid insertion, and there was no involvement of the anterior mass of the brachialis muscles or elbow. We consider that tackler's bony spurs are sometimes, but not often, seen in sumo wrestlers. PMID- 12077672 TI - Rapidly growing fibro-osseous pseudotumor of the digits mimicking extraskeletal osteosarcoma. AB - Fibro-osseous pseudotumor of the digits is a rare benign lesion characterized histopathologically by a fibroblastic proliferation with foci of osseous differentiation. We report a case of fibro-osseous pseudotumor of the digits with a rapidly growing lesion in a 30-year-old woman. Because of its rapid growth and aggressive histopathological appearance, this lesion can be mistaken for a malignant neoplasm such as extraskeletal or parosteal osteosarcoma, despite its benign clinical behavior. For a soft tissue mass in the digit with a clinicopathological feature of pseudomalignancy, the clinician should consider fibro-osseous pseudotumor of the digits as a possible diagnosis, thereby avoiding inappropriately aggressive surgery. PMID- 12077673 TI - Traumatic axial dislocation of the carpus: a case report of transscaphoid pericapitate transhamate axial dislocation. AB - Traumatic axial dislocation of the carpus in a 20-year-old man is described. This injury was accompanied by a crushing injury to the hand. The disruption pattern was different from those of previously reported cases. Despite the restoration of painless wrist motion postoperatively, grip strength remained below normal. Early accurate reduction, fixation, and range of motion (ROM) exercise are the treatment of choice in such complex injuries. PMID- 12077674 TI - Unusual double clavicle fracture complicated by ipsilateral scapular neck fracture. AB - An unusual case of fractures of the mid- and distal clavicle complicated by a scapular neck fracture and second rib fracture on the ipsilateral side is reported. Unstable shoulder girdle or floating shoulder is an unusual injury. The relevant medical literature and treatment of this disorder are discussed. PMID- 12077675 TI - Low-intensity electromagnetic and mechanical modulation of bone growth and repair: are they equivalent? AB - Signals from both electromagnetic fields (EMF) and ultrasound (US) have a clinically significant effect upon bone repair. Both modalities are now a common part of the orthopedist's armamentarium for the care of delayed union, nonunion, and fresh fractures. Dynamization or controlled weight bearing also enhances bone repair. Consideration of the dosimetry of both EMF and US modalities suggests a possible unifying mechanism for the bioeffects from EMF, US, and strain-generated potentials (SGP) signals based on the time-varying electric field, E(t), associated with each type of stimulus. The E(t) field is directly induced with EMF devices and indirectly induced via the streaming potentials associated with the mechanical movement of ionic fluids within bone caniculi or directly past cell surfaces from US and SGP signals. It is shown that both electrically and mechanically induced E(t) have common waveform characteristics at the treatment site and thus can deliver similar doses of electrical stimulation. It is proposed that the time-varying endogenous electric field, E(t), from a time-varying change in the mechanical environment of healing or remodeling bone, can act as a dose dependent growth stimulus. Thus, the primary messenger affecting cellular activity is E(t), suggesting that bone repair or remodeling may be interchangeably modulated using mechanical (including US) or electromagnetic signals. PMID- 12077676 TI - [Problems recognizing pain in geriatrics]. AB - OBJECTIVE: Elderly people often present substantial problems in pain assessment. The aim of the study was to quantify and discuss the problem of recognizing pain in hospitalized geriatric patients. METHODS: 124 geriatric patients and their treating doctors were questioned about pain, pain intensity and influence of pain on daily living in a cross-sectional-study. The score of the Barthel index (activity of daily living, ADL) at admittance and discharge, the cognitive status (monumental status of Folstein, MMS), the use of analgetic drugs and the medical diagnoses were assessed. The answers of the question "Did you/your patient have pain in the past seven days?" from patients and doctors were used to make seven groups. RESULTS: 20 (16,1%) patients could not answer the question about pain. In comparison to the group of patients who were able to answer, they were restricted in ADL and MMS. However, the answers of doctors about pain did not differ. 25 (20,3%) answers of treating doctors were wrong compared to the answers of the patients. In these cases, intensity and influence of pain on daily living were estimated low by the patients. In 14 (11,3%) patients, doctors could not state about pain. These patients were least restricted in ADL at admission and discharge. Altogether, the pain of 13/46 (28,3%) patients was treated insufficiently at the day of the interview. CONCLUSIONS: In half of the geriatric patients, pain was not well or not at all recognized by the treating doctors. Pain of not communicative patients was assessed similarly by the doctor as that of patients who were able to give information. In this geriatric hospital, little support at admittance and a superior improvement in the activities of daily living during the in-hospital treatment lead to less attention to pain by the doctor. The importance of pain therapy in geriatrics is illustrated by the fact that almost every third patient with pain is treated insufficiently at the day of the interview. PMID- 12077677 TI - [Primary headache in childhood and adolescence. The association between changes in family interaction patterns and therapy success--a pilot study]. AB - INTRODUCTION: Family interaction patterns are often involved in diseases and disorders in childhood and adolescence in complex ways (e.g in their development, maintenance and cure). The present study deals with the role of family factors in success in a pediatric headache therapy consisting of group hypnotherapy and systemic family consultation. METHODS: A sample of 12 outpatients, aged 9-15 years and balanced in sex, is investigated. Patients were diagnosed by IHS criteria. Global symptom strain was measured by numeric rating scale (NRS) at pre appointment and at 9-months follow-up appointment. Also family interaction patterns associated with the occurrence of headache symptoms were measured by content analysis. RESULTS: We found an association between changes in two independently assessed variables: global symptom strain and family interaction patterns. (1) When patients assessed global symptom strain as unchanged, family interaction patterns associated with headache were also assessed as unchanged by observers; (2) when patients assessed their global symptom strain as positively changed, family interaction pattern associated with headache were also assessed as positively changed by observers. CONCLUSION: These data provide empirical evidence about when to include family in treatment of pediatric headache: when rigid family interaction patterns associated with headache complicate a symptom change. PMID- 12077678 TI - [Morphine and alternative opioids in cancer pain: the EAPC recommendations]. AB - An expert working group of the European Association for Palliative Care (EAPC) has revised and updated its guidelines on the use of morphine in the management of cancer pain. The revised recommendations presented here give guidance on the use of morphine and the alternative strong opioid analgesics which have been introduced in many parts of the world in recent years. Practical strategies for dealing with difficult situations are described presenting a consensus view where supporting evidence is lacking. The strength of the evidence on which each recommendation is based is indicated. PMID- 12077679 TI - [Guidelines in pain treatment--methodical quality of guidelines for treatment of pain patients]. AB - The committee for quality assurance of the German IASP chapter (DGSS) evaluated all relevant guidelines concerning pain treatment. Quality of guidelines was analysed according to the checklist "Methodical quality of guidelines" by Ollenschlager and the user manual released by the German Medical Centre for Quality Assurance. The guideline for the treatment of back pain released by the German Medical Association was examined as well as the one released by the German Association for physical therapy and rehabilitation, the guideline on cervical and lumbal nerve root compression syndrome of the German Association of Neurosurgeons, the guideline for cancer pain of the Drug Committee of the German Medical Association was compared with the one of the German Interdisciplinary Association for Pain Treatment. The guideline for the treatment of chronic headache and facial pain of the Medical Association was evaluated and the guideline for the treatment of trigeminal neuralgia of the German Association of Neurosurgery and Neurology, also the guideline for the treatment of acute postoperative and posttraumatic pain. All guidelines show deficiencies in different aspects and of different severity. At first there are deficiencies in interdisciplinary formulation of the guidelines and identification and interpretation of evidence of multimodal pain treatment options. The most prominent deficiency is the lack of implementation and application trials or impulses by all author associations. This way all expenditure on releasing guidelines is given away without improving quality of pain treatment. The authors' recommendation is to adjust to guidelines and, if they are working or not, tell the authors and improve interdisciplinary in pain treatment guidelines. PMID- 12077680 TI - [Chronic pain management under daily clinical conditions]. AB - BACKGROUND: For lack of specialized pain centers patients with chronic pain are referred to medical, neurological or geriatric departments. One third of these patients are dissatisfied with pain management during stay in hospital (patients interviews 1998). To improve this situation, a pain management system was developed and tested under daily clinical conditions. METHODS: If the history taking resulted chronic pain (longer than 3 months), the first step was staging the chronicity of pain by the Mainz Pain Staging System. In case of advanced chronic pain (stages II or III), a comprehensive history-taking of the patient's pain including psychometric tests (ADS, SES, PDI) followed. For each patient we worked out an individual and interdisciplinary therapy. To verify the results, each patient was interviewed on the day before dismissal. RESULTS: 27,8% of our patients in half a year period (n = 381) suffered from chronic pain. By pain management activities we were able to rise significantly the satisfaction of the patients with pain therapy. DISCUSSION: It is necessary to search consequently for chronic pain. Especially elder people do not mention their suffer, often believing, that pain is an unavoidable sign of old age. Concentrating on medical reports alone is not adequate for chronic pain and even can aggravate the condition to a significant extent. Chronic pain can neither be explained nor treated without integrating psychological and social factors. A routine diagnostic- and therapy-practice facilitates an interdisciplinary pain therapy, raises the therapeutic capability of the staff and increases the therapeutic outcome. PMID- 12077681 TI - [Deficits in the psychological care of low back pain patients--Comments on the expertise of the expert committee for the health care system regarding low back pain]. AB - A committee of health specialists examined the demand characteristics and efficiency of German health care services and published an expertise on the misallocation of services that among other subjects also covered the area of low back pain. Although we agree with their principal statements, we like to underscore that the objectives mentioned in the report cannot be achieved without the participation of psychologists. The process of chronicity is determined in a significant way by psychological mechanisms. Psychologists investigated these mechanisms, developed interventions for the prevention and treatment of low back pain on the basis of the results, and tested their efficacy in trials with a high methodological standard. Incorporation of psychological expertise does not only optimise prevention and treatment of low back pain and helps to prevent unnecessary suffering of the afflicted, in addition, it offers a contribution a cost containment. PMID- 12077682 TI - [Prophylactic drug management of migraine]. AB - Migraine prophylaxis with drugs is still an essential part of migraine therapy. This is especially true for those patients with frequent migraines who are in danger of developing drug-induced headaches. Migraine prophylaxis should be taken in consideration in patients who suffer from 7 or more migraine days per months in spite of all non-pharmacological efforts. When choosing a prophylactic drug not only efficacy but tolerability and safety for long-term intake should be considered. Prophylactic drugs used to be classified as drugs of first, second and third choice. According to this step care model treatment was started with a drug of first choice and only in case of lack of efficacy or adverse events a drug of lower choice was selected. Today, in contrast to the traditional step care a stratified care is favored. Treatment is individualized based on an assessment of the patients' medical needs, on comorbidity, the migraine phenotype and most important the individual situation of the patient in life. The paper gives an overview of the efficacy and tolerability of drugs used in migraine prophylaxis. PMID- 12077683 TI - Impact of atrio-biventricular pacing to poor left-ventricular function after CABG. AB - AIMS: The relation between acute postoperative management of epicardial pacing and haemodynamic status in patients with poor left-ventricular function after coronary artery bypass grafting (CABG) demonstrates the importance of synchronous ventricular activation and contraction during the vulnerable early postoperative period. METHODS: in 22 patients (mean age - 69.3 +/- 5.4 years) with poor left ventricular function (ejection fraction 29.8 +/- 4.8), we compared the postoperative haemodynamic parameters between atrio-biventricular, atrio monoventricular and atrial pacing 3 - 24 hours after elective coronary artery revascularisation. Temporary epicardial pacing electrodes were placed on the right atrium and the paraseptal region of the left and right ventricle. The ventricular pacing modus was confirmed by surface electrocardiogram (EGG). We used overdrive rate pacing. RESULTS: In patients with left bundle branch block, atrio-left-ventricular and atrio-biventricular pacing increased cardiac index and decreased wedge pressure. Atrial pacing and atrio-right-ventricular pacing decreased cardiac index. In contrast, atrio-right-ventricular and atrio biventricular pacing increased cardiac index in patients with right bundle brunch block. CONCLUSION: Atrio-biventricular pacing increased cardiac index and decreased wedge pressure compared with AAI pacing. In patients with wall-motion abnormalities and impaired cardiac conduction, a site-specific pacing therapy can help to optimize postoperative haemodynamics and reduce the application of inotropic substances. PMID- 12077684 TI - Elastase release following myocardial ischemia during extracorporeal circulation (ECC) -- marker of ongoing systemic inflammation? AB - BACKGROUND: 'Post-Perfusion Syndrome' (PPS) after cardiopulmonary bypass (CPB) is known to be evoked by inflammatory reactions. The hypothesis of a pathogenetic role for the neutrophil granulocytes in this inflammation would be strengthened if elevated concentrations of a neutrophil product such as elastase could be demonstrated, particularly in case of a PPS or a systemic inflammatory response syndrome (SIRS). METHODS: In a randomized prospective double-blind study, 40 patients undergoing aortocoronary bypass grafting (CABG) were divided into 4 groups of 10 patients each. One group served as the control group, one received prostacyclin (PGl 2 ), the third group was substituted with high-dosed aprotinin and the last group was treated with a combination of PGl 2 and aprotinin. 6 blood samples were taken from every patient perioperatively, and plasma elastase (PE), procalcitonin (PCT), C 1 -esterase inhibitor (CEI) and parameters of coagulation and fibrinolysis were determined. RESULTS: Levels of elastase increased significantly in all intra- and postoperative blood samples compared to the preoperative baseline values (< 30 microg/l, p < 0.05). The elastase release was even more pronounced in the control and aprotinin group (170 +/- 23 microg/l; 175 +/- 14 microg/l during ECC) compared to patients who received prostacyclin (142 = 21 microg/l, p < 0.05). Duration of myocardial ischemia could be directly correlated to elastase levels at the end of CPB. 10 of the 40 patients suffered postoperatively from a PPS or a SIRS; in these patients, elastase levels at the end of CPB were significantly higher (188 +/- 26 microg/l vs. 138 +/- 22 microg/l, p < 0.05). Immediately after the operation, these 10 patients also showed significant changes in the cascades of coagulation and fibrinolysis resulting in a hypercoagulatory state. Levels of PCT and CEI did not change significantly during and after ECC. CONCLUSIONS: Our results indicate that CPB initiates an elastase release that can be suppressed by prostacyclin. Increased intraoperative elastase levels in patients with PPS show that elastase may be an indicator of ongoing systemic inflammation, possibly causing complications due to a hypercoagulatory state. Myocardial ischemia seems to be one reason for this elastase release. It can be speculated that early PGl2-infusion could be a therapeutic option in inflammatory diseases caused by ECC. PMID- 12077685 TI - Circulating microemboli after composite replacement or valve-sparing aortic root surgery. AB - BACKGROUND: The purpose of this study was to determine whether microembolic signals (MES) occur after valve-sparing operations on the aortic root. One of the advantages of these procedures relates to the freedom of macroemboli without anticoagulation. Whether this holds true for circulating microemboli has not yet been verified. METHODS: For comparison, 8 male patients (mean age: 51.8 +/- 12.8 years) were investigated 20.5 +/- 8.4 months after implantation of a mechanical composite graft (group I) and 9 female and 7 male patients (mean age 55.0 +/- 13.4 years) 23.5 +/- 20.0 months after valve-sparing replacement of the aortic root (group II). The middle cerebral artery was insonated for 2 periods of 30 min, breathing room air or O 2 at 9 l/min. RESULT: Breathing room air, the amount of MES was considerably smaller in group II (0.94 +/- 1.95 vs. 56.1 +/- 58.9 per 30 min, p = 0.006). The difference was less pronounced (0.5 +/- 1.3 vs. 28.9 +/- 42.6 per 30 min, p = 0.009) breathing oxygen. Breathing oxygen reduced MES significantly in group I (p < 0.05) but not in group II (p > 0.05). CONCLUSIONS: Aortic valve-sparing operations induce MES at a significantly lower rate than composite aortic valve replacement using a mechanical valve. PMID- 12077686 TI - Influence of intensified medical treatment and organ allocation on outcome of transplant candidates. AB - BACKGROUND: To test how the organization of a pre-transplant clinic and changes in organ allocation modus influence the survival of potential transplant candidates, the survival of patients referred for transplant evaluation between 4/93 and 4/96 (group A) was compared to that of patients referred from 5/96 to 7/00(group B). METHODS: After screening for transplant indication, group A was followed by the referring physician up to transplantation or 3-month reevaluation. Group B was closely followed by a specialized heart-failure clinic. Group A was transplanted according to Eurotransplant criteria, with waiting time being the strongest priority criterion. Due to an allocation partnership off our transplant centers, group B could be transplanted according to medical urgency regardless of waiting time. RESULTS: Overall one-year survival after referral was 69.8% for group A vs. 91 %for group B (p <0.0001). Transplantation within 1 year was required in more group A than group B patients (34% vs. 23%)with worse one year post-transplant survival in group A (82%vs. 93%). CONCLUSIONS: Intensified treatment by a specialized heart failure program and an allocation system that allows for preferred transplantation of the 'sickest' patient improved over-all survival of transplant candidates and reduced the percentage of patients requiring transplantation. PMID- 12077687 TI - Influence of ACE inhibition on myocardial damage, the Kallikrein-Kinin system and hemostasis during cardiopulmonary bypass surgery. AB - BACKGROUND: ACE inhibitors may have a cardioprotective effect by enhancing bradykinin levels during cardiopulmonary bypass (CPB). However, ACE inhibition could lead to unwelcome effects on the kallikrein contact phase during CPB (since reduction of kallikrein activity by aprotinin has been shown to be beneficial) and may alter the hemostasis. We examined the effects of ACE inhibitors on intraoperative myocardial damage, kallikrein contact phase and hemostasis in patients undergoing CPB. METHODS: 47 patients randomly received either 20 mg/d enalapril or placebo. Creatine kinase (CK and CK-MB), lactate dehydrogenase (LDH), troponin T (TnT), thrombin-antithrombin III complex (TAT), fibrinogen and kallikrein-like activity were measured before surgery, during and immediately after CPB, at the end of surgery and 1, 3 and 5 days after surgery. RESULTS: No significant differences between enalapril- and placebo- treated patients concerning CK (318 +/- 38.6 U/l vs. 316 +/- 16.8 U/l), CK-MB, LDH, TnT (1.81 +/- 0.45 ng/ml vs. 1.52 +/- 0.34 ng/ml), TAT, fibrinogen and kallikrein-like-activity could be found during study period. CONCLUSIONS: Reduction of ischemic injury during CPB is not achieved with ACE inhibitors. However, treatment of patients with ACE inhibitors before and during CPB is fully feasible without side effects affecting the kallikrein contact phase or significant influence on hemostasis. PMID- 12077688 TI - Left atrial radiofrequency compartmentalization for chronic atrial fibrillation during heart surgery. AB - BACKGROUND: The excellent results of the Maze III operation have demonstrated that a surgical cure of atrial fibrillation (AF) is possible. A simplified Maze procedure with radiofrequency (RF)ablation of the atrial tissue added to open heart surgery may help to cure chronic AF with low risk. METHODS: From May 1998 to March 2001, an RF left atrial compartmentalization concomitant to a cardiac surgical intervention was performed on 40 of 1,258 patients (3.2 %). Thirty-seven patients had mitral valve disease, two had aortic valve disease and one had coronary artery disease. All patients presented with chronic refractory AF for a mean time of 40.4 months ranging from 4 months to 18 years. RESULTS: Mitral valve replacement (MVR) was performed in 19 patients, mitral valve repair in 12, combined aortic and mitral valve procedures were performed in 6, aortic valve replacement(AVR) in 2 and CABG in 1 patient. Cardiopulmonary bypass time for the complete procedure was 138 +/- 32 minutes with an ablation time of 19 +/- 5 minutes. No complication related to RF application was noted. Sinus rhythm was present in 68% after 12.5 +/- 8.5 months. CONCLUSIONS: RF left atrial compartmentalization combined with cardiac surgical interventions proved to be easy and safe to perform. The long-term results still have to be confirmed with further regular patient follow-up examinations. PMID- 12077689 TI - Endoscopic saphenous vein harvesting for CABG -- a randomized, prospective trial. AB - BACKGROUND: The saphenous vein is an established conduit for coronary revascularization. Disadvantages of traditional harvest technique are significant pain and morbidity. We compared the endoscopic harvest technique with the traditional method. METHOD: 140 coronary artery bypass graft (CABG) patients were randomized into 2 groups: endoscopic vein harvesting (EVH; n = 80) and traditional open vein harvesting (OVH; n = 60). Analysis included preoperative risk factors for wound complication, harvesting time, graft injury, and intraoperative and postoperative complications. Patient follow-up lasted 3 months. RESULTS: The preoperative risk profiles of the groups were comparable. In the EVH group, 5 patients (7.1 %) had to be switched to the open technique. EVH time was 45 +/- 6.2 min vs. 31.1 +/- 6.5 min. Two patients (2.5 %) had to be revised because of bleeding complication vs. 6 (10 %) in the OVH group. No local infections or wound complications were observed in the EVH group vs. 11 (18 %) cases in the OVH group. Two OVH cases (3.6 %) were readmitted for wound debridement. All EVH patients reported less pain and were completely satisfied by the cosmetic results. CONCLUSION: EVH is a safe and efficient technique for CABG. Morbidity was significantly lower, with reduced pain and better cosmetic results. EVH time was significantly longer compared to the traditional harvesting technique. PMID- 12077690 TI - Beta-blockade in 200 coronary bypass grafting procedures. AB - BACKGROUND: Continuous perfusion of the coronary arteries with high-dose beta blocker (esmolol)-enriched blood has been shown to represent an alternative for myocardial protection during coronary bypass grafting (CABG). Here, we will report on our experience in 200 unselected consecutive cases where this technique was used. METHODS: Eighty percent of the patients (age: 63.3 +/- 0.6 years, ejection fraction: 60 +/- 1.2 %, emergency cases: 11 %) had 3-vessel disease, 34 % had a history of myocardial infarction within less than 90 days preoperatively. The Euro score amounted to 6 +/- 0.3. During 52 +/- 1.2 min of aortic cross-clamp time on normothermic cardiopulmonary bypass, 2.9 +/- 0.1 distal anastomoses were performed on a slow hypocontractile beating heart induced by continuous infusion of 788 +/- 20 mg esmolol per operation. All data: mean +/- SEM. RESULTS: Postoperatively, patients were ventilated for 25 +/- 5.1 hours and stayed on ICU for 2.3 +/- 0.3 days. The postoperative myocardial infarction rate was 4%. Patients left the hospital after 11.2 +/- 0.4 days. Thirty-day mortality was 2.5 %, and 3-month mortality was 3 %. CONCLUSIONS: High-dose beta-blockade is a safe and effective technique in CABG procedures. It may be especially advantageous in high-risk patients. PMID- 12077691 TI - Management of iatrogenous esophagus perforation. AB - OBJECTIVES: The management of patients with iatrogenous, instrumentally caused esophageal disruptions is still a controversially discussed challenge for any physician. We reviewed indications, morbidity, and mortality of esophagectomy compared to less aggressive treatment schemes in the light of our single-unit experience. PATIENTS AND METHODS: We reviewed the cases of 38 consecutive patients suffering from iatrogenous esophageal disruption treated within the last 10 years at the University Medical School Graz. RESULTS: Severe sepsis was diagnosed in seventeen patients; fourteen of the seventeen patients were treated by esophagectomy followed by primary or secondary reconstruction at a mortality rate of 28.6 % (four of fourteen patients). All fourteen patients thus treated had major preexisting esophageal pathologies. In three of the total seventeen cases reviewed, conservative treatment modalities were used. The mortality rate in the non-septic group treated by direct suturing (n = 21) was 4.8 % (one of twenty-one patients). Major preexisting esophageal pathology was present in thirteen. The overall mortality was 13.2 % (five of thirty-eight patients). CONCLUSION: The strategy of primary repair for iatrogenous esophageal injury should only be adopted in patients with minor or without intrinsic esophageal disease, and in the absence of severe sepsis suggesting mediastinitis. Severe sepsis following iatrogenous esophageal trauma should prompt the decision for esophagectomy where anatomically and/or oncologically possible. PMID- 12077692 TI - The value of mediastinoscopy in preoperative evaluation of mediastinal involvement in non-small-cell lung cancer patients with clinical NO disease. AB - BACKGROUND: The efficacy of mediastinal lymph node examination using cervical mediastinoscopy in operable non-small cell lung cancer patients without radiological nodal involvement on computerized tomography (CT) has been elusive. METHODS: The value of mediastinoscopy as a staging modality for assessing the mediastinal lymph node status was evaluated in 79 patients with presumed resectable non-small-cell lung cancer (NSCLC) with mediastinal nodes smaller than 1 cm (NO) form the CT scan. Sixty-one patients who did not have nodal involvement at mediastinoscopy and had complete medical records underwent complete resection. RESULTS: Negative predictive value (NPV) of the CT scan according to mediastinoscopy was 92.4 %. Histopathological examination of the surgical specimen showed the NPV of mediastinoscopy to be 93.4 %. Only 4 patients (3 patients with N2, 1 patient with N3 disease) were not correctly staged using CT scanning and mediastinoscopy. According to the pathological examination, the NPV of CT was found to be lower (76.5 %) in patients with adenocarcinoma, but the difference was not statistically significant (p > 0.128) CONCLUSION: Although the likelihood of surgical-pathological N2 is slightly higher in patients with adenocarcinoma, radiological examination of patients with cNO NSCLC disease can be as accurate as mediastinoscopy in appropriately staging mediastinal lymph node involvement. PMID- 12077693 TI - A particular case with long-term follow-up of rare malignant hemangiopericytoma of the lung with metachronous diaphragmatic metastasis. AB - The authors report the case of a woman with a primary hemangiopericytoma of the lung and successive metachronous diaphragmatic metastasis treated with surgery only according to the patient's decision. The patient was first submitted to left lower lobectomy with resection of diaphragm and three ribs en-bloc for pulmonary sarcoma. One year later, a chest computed tomography (CT) scan showed a small lesion above the diaphragm on the left side. She refused the proposed operation and returned fifteen months later with a new thorax CT, and told us that she had in the meantime become pregnant and given birth. A rethoracotomy to remove the metastatic diaphragmatic tumor was performed. Chemotherapy with anthracycline and iphosphamide was proposed, but the patient again refused therapy for fear of side effects impairing another pregnancy. Currently, the patient is alive and disease free, 68 months after the first treatment. The authors emphasize the long-term follow-up obtained with surgery alone in this kind of rare sarcoma. PMID- 12077694 TI - Delayed type A dissection after primary successful interventional treatment with endoluminal stent graft of the descending thoracic aorta. PMID- 12077695 TI - Modified pericardial closure to protect internal mammary artery grafts in coronary artery bypass. AB - Redo operations after coronary artery bypass surgery have been on the rise, and myocardial and graft injury during resternotomy a catastrophe. Closure of pericardium after a traditional midline incision may lead to graft distortion. In this report, we will describe a technique of pericardial closure and thymus coverage to protect bilateral internal mammary artery grafts from damage. PMID- 12077696 TI - Tissue engineering of heart valves -- current aspects. AB - Tissue engineering of heart valves is an evolving research field. Driven by the shortcomings of the heart valve substitutes currently available, such as need for anticoagulation, susceptibility to infections, inability to grow and autorepair, the multidisciplinary approach for designing and growing viable heart valves identical to the native heart valves has begun. The following will give an update of the recent developments, current limitations and potential future applications of tissue-engineered heart valves. PMID- 12077699 TI - Considerations in metatarsalgia and midfoot pain: an MR imaging perspective. AB - There are many conditions that have the potential to produce the symptom of pain in the metatarsal and midfoot regions. Pathologic processes that cause pain include post-traumatic disorders, circulatory disorders, arthritides, neuroarthropathy, and conditions that result in a biomechanical imbalance in the foot. Magnetic resonance (MR) imaging is a powerful imaging tool since it provides a noninvasive method for diagnosis of different foot disorders. In this article, we will discuss the MR manifestation of a number of pathologic entities that elicit foot pain as a primary symptom. PMID- 12077700 TI - MR imaging of abnormalities of the plantar fascia. AB - Foot pain and disability may curtail the performance of simple routine tasks, work-related duties, and athletic activities. Disorders of the plantar fascia are commonly encountered in the diagnostic evaluation of subcalcaneal heel pain. The approach to imaging the ankle and foot has undergone significant amendment with the introduction of magnetic resonance (MR) imaging. MR imaging allows direct visualization of the plantar fascia and permits a comprehensive evaluation of a wide spectrum of disorders involving the fascia and perifascial structures, including bones, ligaments, musculotendinous units, and intrinsic muscles of the foot. The objectives of our cadaveric and clinical study are to describe the normal MR imaging appearance of the plantar fascia and analyze the signal intensity changes in the fascia and perifascial tissue with conventional MR imaging in symptomatic patients. We present a brief overview of common disease processes involving the plantar fascia that may prove practical for routine imaging evaluation of patients with suspected pathology. PMID- 12077701 TI - MR imaging of the tendons of the foot and ankle. AB - Hindfoot pain from tendon pathology is common and seen in a wide range of patients from young athletes to older sedentary individuals. Magnetic resonance (MR) is an excellent technique for imaging tendons and for identifying injuries that can be treated with surgery. MR also demonstrates the presence of bone marrow edema, which is a cause of pain and often a marker for adjacent tendon injury. Finally, MR can reveal other etiologies that cause similar hindfoot pain, such as osteochondral injuries and impingement. In this article, we review the normal MR appearance of the ankle tendons and tendon sheath fluid. MR protocols for imaging the various ankle tendon groups are presented. We discuss the MR appearance of tendon pathology, including tendonosis/ tendinopathy, tenosynovitis and peritendonosis, partial and complete tears, subluxation and dislocation, and entrapment. The Achilles, medial, lateral, and anterior tendon groups are discussed separately so as to focus on the MR appearance specific to each region of the ankle. PMID- 12077703 TI - MR imaging of soft-tissue masses of the foot. AB - As with other parts of the musculoskeletal system, the soft tissues of the foot can be affected by a wide variety of pathologic entities including trauma, congenital abnormalities, infections, and neoplastic disorders. While plain radiographs are usually the initial examination for evaluation of pathology, magnetic resonance imaging (MRI) is critical to evaluate for abnormalities within the ligaments, tendons, and other nonosseous structures within the foot. The constellation of clinical and MRI findings often allows a relatively specific diagnosis to be rendered. This article discusses both benign and malignant processes within the soft tissues of the foot and presents their characteristic imaging findings with MRI. PMID- 12077702 TI - Ankle impingement syndromes. AB - The term "ankle impingement" encompasses a broad range of conditions that are typically post-traumatic and often chronic. Various forms of mechanical impingement can result from synovial proliferation, bone spur formation, or ligamentous scarring and hypertrophy. Since symptoms and physical findings can mimic a variety of disorders, accurate diagnosis may remain elusive, and proper effective therapy may be delayed. The objective of this article is to define and elucidate the etiology of the various forms of ankle impingement, clarify the range of associated osseous and soft-tissue pathology, and describe the imaging features and therapeutic options. PMID- 12077704 TI - Painful accessory bones of the foot. AB - Sesamoid bones and secondary centers of ossification account for the accessory bones of the foot, and there is a wide range of frequency with which these different ossicles appear. They are usually incidental asymptomatic findings on imaging studies, and their clinical importance lies in not mistaking them for fractures. Occasionally these ossicles can become painful due to fracture, degenerative changes, avascular necrosis, and irritation or impingement of adjacent soft tissue. PMID- 12077705 TI - Ultrasound-guided interventions in the foot and ankle. AB - Both chronic as well as acute foot and ankle pain are common indications for visits to general practitioners, rheumatologists, and orthopedic surgeons. The causes for foot and ankle pain are varied and can include osseous pathology (fractures), yet the far greater majority are secondary to soft-tissue injuries and inflammation. Regional corticosteroid injections, traditionally performed using anatomic landmarks, can be inaccurate and miss intended targets. The use of ultrasound for guidance for interventional radiologic procedures is well known, including guidance for vascular as well as visceral interventions. Using sonography to guide for interventions in the musculoskeletal system, specifically the foot and ankle, yields accurate placement of the needle tip and subsequent anesthetic/steroid injection as well as diagnostic aspiration of tendon sheaths, joint spaces, and bursae. PMID- 12077706 TI - Mutations in the cone photoreceptor G-protein alpha-subunit gene GNAT2 in patients with achromatopsia. AB - Achromatopsia is an autosomal recessively inherited visual disorder that is present from birth and that features the absence of color discrimination. We here report the identification of five independent families with achromatopsia that segregate protein-truncation mutations in the GNAT2 gene, located on chromosome 1p13. GNAT2 encodes the cone photoreceptor-specific alpha-subunit of transducin, a G-protein of the phototransduction cascade, which couples to the visual pigment(s). Our results demonstrate that GNAT2 is the third gene implicated in achromatopsia. PMID- 12077707 TI - Influence of human leukocyte antigen-DRB1 on the susceptibility and severity of rheumatoid arthritis. AB - BACKGROUND AND OBJECTIVES: All human leukocyte antigen (HLA)-DRB1 alleles associated with rheumatoid arthritis (RA) encode a conserved amino acid sequence (QKRAA, QRRAA, or RRRAA) at position 70-74 in the third hypervariable region (HVR3) of the DRbeta(1) chain, which is commonly called the shared epitope (SE). Several studies, however, have associated the HLA-DRB1 gene in RA severity and progression rather than with susceptibility. Moreover, the association with disease severity and presence of the SE varies among different ethnic populations. HLA-DRB1 alleles also influence the disease onset. In this manuscript, the role of the HLA genes in RA was examined. METHODS: A retrospective review of the literature was conducted to analyze the influence of the HLA-class II genes on the susceptibility, severity and protection against RA. RESULTS: The HLA-DRB1*0401/*0404 genotype was associated with a higher risk for early disease onset in more severe forms in patients from the United Kingdom (UK). In northwest Spain, RA onset under 40 years is strongly associated with HLA DRB1*0401 and *0404. In contrast, RA onset above 60 years is associated with HLA DRB1*01. The protection against RA linked to some HLA-DRB1 alleles encoding a DERAA sequence of amino acids at position 70-74 in the HVR3 of the DRbeta1 chain, and specifically aspartic acid (D) at position 70 of this chain, recently was confirmed in both UK and northwest Spanish populations. Besides HLA-class II, other genes may be implicated in RA. Polymorphism in the tumor necrosis factor (TNF) region seems to be associated with RA, even in patients without the HLA DRB1 SE. However, other genes such as interleukin-1 (IL-1) and corticotropin releasing hormone may play a role in susceptibility to RA. CONCLUSIONS: The additive effect of various genes may account for the development of RA and its clinical severity. PMID- 12077708 TI - Shared epitopes and rheumatoid arthritis: disease associations in Greece and meta analysis of Mediterranean European populations. AB - OBJECTIVES: To assess the strength of the associations between HLA shared epitopes (SE) and rheumatoid arthritis (RA) susceptibility, articular disease severity, and extra-articular features in Mediterranean European populations. METHODS: One hundred and seventy-four Greek RA patients and 103 controls were evaluated. Data were then included in a meta-analysis of 9 studies of Mediterranean European populations (959 RA patients and 1,405 controls). RESULTS: In our study population, SE alleles were significantly more common in RA patients than in controls (odds ratio [OR], 2.5; 95% confidence interval [CI], 1.4-4.3). Larsen radiologic score was predicted by SE and disease duration. SE did not increase the risk of any extra-articular manifestation. The meta-analysis showed a pooled OR of 3.7 (95% CI, 2.6-5.2) for susceptibility to RA conferred by SE (OR, 3.4 v 3.9 in Greek v non-Greek populations). CONCLUSIONS: SE determine articular destruction without increasing the risk of extra-articular manifestations. The immunogenetic associations of RA susceptibility are consistent, but their strength may depend on the SE prevalence in different ethnic groups. PMID- 12077709 TI - Genotype/phenotype correlations in Arab patients with familial Mediterranean fever. AB - OBJECTIVES: To study the phenotype/genotype correlations in Arab patients with familial Mediterranean fever (FMF). PATIENTS AND METHODS: The study was performed in a 3-year period (February 1998-February 2001). Patients were seen in the pediatric FMF clinic of Jordan University Hospital, and the diagnosis of FMF was made according to published criteria. Screening for mutations was carried out by direct sequencing of the entire coding sequence of exon 10 and its donor splice site and by restriction endonuclease testing for mutations in exon 2. A total of 278 patients with clinically positive FMF were screened. RESULTS: Of the 278 patients, 50 (18%) had 2 mutations identified, and 76 (27%) other patients had only 1 mutation identified. The 50 patients with 2 mutations are the subject of this report. The M694V/M694V and the M694V/V726A and M694I/M694I genotypes were the most common (30%, 16%, and 14%, respectively). Three homozygous genotypes (M694V/M694V, V726A/V726A, and M694I/M694I) and 2 compound heterozygous genotypes (M694V/V726A and V726A/M680I) accounted for 78% of mutations. The difference in the mean severity score (14 +/- 2) of the M694V/M694V group and the V726A/V726A (mean severity score, 10 +/- 3) and M694I/M6941 (mean severity score, 6 +/- 1) groups was statistically significant (P =.003 and.0, respectively). The difference between the M649V/M694V group and the M694V/V726A (mean severity score, 15 +/- 2) was not statistically significant (P = 0.31). CONCLUSIONS: The genotypes M694V/M694V and M694V/V726A have a severe clinical course in Arab patients with FMF, whereas the M694I/M694I is associated with mild disease. PMID- 12077710 TI - Polyarteritis nodosa presenting as polymyositis. AB - BACKGROUND: Skeletal muscle involvement has been well documented in patients with polyarteritis nodosa (PAN), and symptoms referable to skeletal muscle are not uncommon. However, polymyositis as a mode of presentation of PAN is uncommon. This unusual presentation of PAN has been reported only once previously in the English literature. OBJECTIVE: This study describes a patient who had diffuse weakness, myalgias, and markedly elevated serum creatinine phosphokinase, mimicking polymyositis. The literature dealing with the clinical aspects of muscle involvement in PAN is reviewed. RESULTS: A 24-year-old man was admitted to the hospital with a 1-month history of fever, myalgia, and muscle weakness. Necrotizing vasculitis was shown on subsequent muscle biopsy, consistent with PAN. Literature review indicated that muscle involvement is common in PAN, as has been shown by the frequency of muscular symptoms and by histologic evidence obtained from both clinical and autopsy studies. Nineteen percent of patients with PAN had documented myopathy, and autopsy series have shown skeletal muscle involvement in 30% to 48% of cases. However, polymyositis as a mode of presentation of PAN is rare. We found only 1 other patient with PAN who had elevated creatinine phosphokinase and diffuse myopathy suggestive of polymyositis. CONCLUSIONS: PAN should be suspected in cases of focal or diffuse myopathy, especially in the context of a systemic disease. Biopsy of symptomatic muscles or EMG-directed biopsies can be helpful in establishing a diagnosis of PAN to allow the physician to provide early treatment. PMID- 12077711 TI - Relapsing polychondritis: a clinical review. AB - OBJECTIVE: This study comprehensively reviews the literature related to relapsing polychondritis (RP). METHODS: A detailed search via MEDLINE (PubMed) was performed using relapsing polychondritis as the key term. Relevant articles were analyzed with a focus on history, epidemiology, etiology, pathogenesis, clinical manifestations, diagnosis, treatment, and prognosis of RP. RESULTS: RP is a rare episodic and progressive inflammatory disease of presumed autoimmune etiology first described in 1923. RP affects cartilage in multiple organs, such as the ear, nose, larynx, trachea, bronchi, and joints. In addition, it can affect proteoglycan-rich tissues, such as the eyes, aorta, heart, and skin. The diagnosis of RP is based on the presence of clinical criteria. A standardized therapeutic protocol for RP has not been established. Nonsteroidal anti inflammatory drugs, dapsone and/or colchicine, may control disease activity in some patients. In other patients, immunosuppressive drugs and prednisone have been effective. RP is a potentially lethal disease; pulmonary infection, systemic vasculitis, airway collapse, and renal failure are the most common causes of death. Earlier studies indicate survival rates between 70% at 4 years and 55% at 10 years. In a recent study, a survival rate of 94% at 8 years may be due to improved medical and surgical management. CONCLUSIONS: RP is a rare, multisystemic, and potentially fatal disease. The pathogenesis and optimal therapeutic approach to patients with RP is poorly understood. PMID- 12077712 TI - TAP, HLA-DQB1, and HLA-DRB1 polymorphism in Colombian patients with primary Sjogren's syndrome. AB - OBJECTIVE: Although primary Sjogren's syndrome (pSS) has a worldwide distribution, little data is available on pSS immunogenetics in non-white populations. Thus, we investigated the influence of transporters associated with antigen processing (TAP), human leukocyte antigen (HLA)-DQB1, and HLA-DRB1 gene polymorphism in mestizo Colombian patients with pSS. METHODS: In this cross sectional and controlled study, all patients met the European criteria for classification of pSS. TAP and HLA typing was performed by polymerase chain reaction techniques. Genetic data analysis was performed to detect deviations from the expected Hardy-Weinberg (H-W) proportions and to determine the presence of population stratification or subdivision and the existence of linkage disequilibrium between pairs of loci. RESULTS: Seventy-three Colombian patients with pSS (95% women) and 76 healthy controls were studied. Although significant associations were not observed between TAP or HLA polymorphism and disease, strong linkage disequilibrium among the loci TAP2 and DQB1 was found in patients. Deviations from the H-W expected value were found in the DQB1 locus of patients (P =.02). HLA-DRB1*0301-DQB1*0201 haplotype was associated with more severe histopathologic disease (odds ratio [OR], 15.5; 95% confidence interval [CI], 1.9 129; P =.001) and the presence of anti-Ro (OR, 3.8; 95% CI, 1-15; P =.04) and anti-La antibodies (OR, 4.3; 95% CI, 1.3-14; P =.01). CONCLUSION: The data show genetic evidence suggesting that, in Colombians, a region immersed or in the vicinity in the HLA class II system is strongly associated with a predisposition to acquire pSS, which is probably located between the TAP2 and HLA-DQB1 locus. Our results confirm that the HLA-DRB1*0301-DQB1*0201 haplotype participates in the pathogenesis of pSS. PMID- 12077713 TI - Abdominal scintigraphy using technetium Tc 99m hexylmethylpropylene amine oxime labeled leukocytes in patients with seronegative spondyloarthropathies. AB - OBJECTIVE: To assess the potential benefits of technetium Tc 99m hexylmethylpropylene amine oxime (Tc 99m HMPAO)-labeled leukocyte scintigraphy in a group of patients with spondyloarthropathies (SpAs), overt gastrointestinal symptoms, and negative extensive endoscopic/radiologic test results. PATIENTS AND METHODS: Ten patients with SpAs and overt gastrointestinal symptoms were included in this study. All patients underwent colonoscopy and small bowel barium studies, and results were negative. Abdominal scintigraphy with Tc 99m HMPAO-labeled leukocytes was performed in all the patients. Clinical and laboratory data and response to treatment was recorded. RESULTS: The Tc 99m HMPAO-labeled leukocyte scintigraphy was positive in 5 of 10 patients, demonstrating uptake at the terminal ileum which is very suggestive of Crohn disease. The 5 scintigraphically positive patients were treated with sulfasalazine (SSZ). Four patients responded to SSZ with significant improvement of both gastrointestinal and joint symptoms. CONCLUSIONS: In 5 of 10 patients with SpA and suspected inflammatory bowel disease on clinical grounds, evidence of inflammatory bowel disease was shown by scintigraphic studies in which conventional invasive procedures failed. Tc 99m HMPAO-labeled leukocyte scintigraphy should be considered in the evaluation of patients with SpA. PMID- 12077714 TI - Psoriatic spondyloarthropathy: a comparative study between HLA-B27 positive and HLA-B27 negative disease. AB - OBJECTIVES: To evaluate the relative contribution of the human leukocyte antigen (HLA)-B27 to psoriatic spondyloarthropathy (PsSpA) susceptibility and to analyze whether this antigen contributes to disease expression. METHODS: This cross sectional study included 70 patients (mean age 48 +/- 14.5 years; 44 men and 26 women). PsSpA was defined according to radiological findings (grade 2 or more sacroiliitis), and patients were classified into 3 main subtypes: isolated axial disease (n = 16), axial plus oligoarthritis (n = 29) and axial plus polyarthritis (n = 25). All patients were studied following a standard protocol that included the collection of demographic and epidemiological data, clinical history, radiographs, complementary tests, physical examination, and HLA-B27 testing (serological method). For functional evaluation, the Health Assessment Questionnaire-Specific for spondyloarthropathy (HAQ-S) was used. Patients with and without HLA-B27 antigen were compared on the basis of the data. RESULTS: Twenty-four patients (34%) carried the HLA-B27 antigen (RR 6.4, P <.0004). Fifty six percent of those patients with the isolated axial pattern had this antigen, compared with 24% in the poly-arthritis axial pattern and 31% of those in the oligo-arthritis axial group (P =.016). Univariate analysis demonstrated correlations between HLA-B27 and an earlier age of onset for both psoriasis (P =.028) and arthritis (P =.006), male gender (P =.002), bilateral sacroiliitis (P =.002), and uveitis (P =.026). HLA-B27 negative patients developed more peripheral erosions than HLA-B27 positive patients (P =.05). No correlation was found between B27 and clinical symptoms of back involvement, syndesmophytes, or functional impairment. CONCLUSIONS: The HLA-B27 antigen is not only important for PsSpA susceptibility, but also determines some clinical features. This antigen was associated with earlier age of psoriasis and arthritis onset, bilateral sacroiliitis, and male gender. However, it was not associated with either the severity or extension of the spondylitic process or with functional impairment. PMID- 12077715 TI - Dyslipidemia and the vulnerable plaque. AB - In the last decade, an increasingly sophisticated understanding of the pathogenesis of atherosclerosis and its cardiovascular consequences has emerged. The characteristics of the unstable atherosclerotic plaque, the substrate for the majority of acute coronary events, have been well defined: mild-to-moderate stenosis, a lipid-rich pool, few smooth muscle cells, a friable fibrous cap, and macrophage infiltration. Lipid modification, an important cardiovascular risk reduction strategy, induces a number of effects at the vascular level that may contribute to the clinical benefits seen in large-scale, prospective prevention trials. New developments in imaging technologies may afford improved opportunities to visualize the at-risk plaque and may provide new insights into the optimal management of the unstable plaque. PMID- 12077717 TI - Accelerated atherosclerosis in saphenous vein bypass grafts: a spectrum of diffuse plaque instability. AB - Our understanding of plaque instability may be extended to vein graft atherosclerosis, which appears to represent the end of a continuum of plaque instability. Compared with plaque in native coronary arteries, vein graft atheroma is more diffuse and vulnerable to rupture, and the consequences of plaque rupture in vein grafts seem to be associated with almost certain thrombotic occlusion within 7 to 12 years after surgery. PMID- 12077716 TI - Plaque instability in peripheral vessels. AB - Vulnerable atherosclerotic plaques, those that are prone to rupture, are soft and lipid rich. Other factors that contribute to destabilizing these plaques are inflammation and vasoconstriction. Although less information is available regarding noncoronary vulnerable plaques than the more frequently studied coronary plaques, it appears that the amount of soft lipid "gruel" as well as in situ inflammation plays a crucial role in destabilizing peripheral vascular atherosclerotic plaque, eventually causing its rupture. As in the coronary circulation, it is likely that the majority of acute plaque rupture events are clinically silent. Nevertheless, preventing plaque rupture in the noncoronary circulation is of the utmost importance because this is the presumed mechanism of progression of peripheral vascular atherosclerotic lesions. PMID- 12077718 TI - Multifocal coronary plaque instability. AB - Recent observations document that many patients with acute coronary syndromes harbor multiple complex plaques by angiography, which correlate with multiple plaque ruptures and clots at necropsy. Multifocal plaque instability is evident not only in coronary vessels but also in peripheral vessels where peripheral and coronary plaque instability may exist concomitantly. These observations support the concept that plaque instability is not merely a local vascular accident but instead reflects more systemic pathophysiological processes with potential to destabilize atherosclerotic plaques throughout the cardiovascular system. PMID- 12077720 TI - Systemic and local inflammation in patients with unstable atherosclerotic plaques. AB - The response to injury in the vasculature and the heart is inflammation. Atherosclerosis is often the result of injury followed by inflammation and atherosclerosis. Vascular and myocardial infections from various pathogens, including viruses, bacteria, chlamydia, and other infections result in vascular inflammation and almost certainly play a role in the development of atherosclerosis and acute coronary heart disease syndromes in at least some patients. Current evidence favors prior exposure to multiple pathogens as most likely playing a role in initiating inflammation and contributing to atherosclerosis. Genetic predisposition is almost certainly an important factor in the development of inflammation, impaired endothelial vascular repair, vascular infection, thrombosis, and atherosclerosis. The aging process itself is most likely associated with altered vascular and myocardial defense mechanisms predisposing to inflammation. The oxidation of cholesterol and low-density lipoprotein (LDL) leads to the production of oxidized radicals that promote vascular inflammation. Interventional injury, including angioplasty and stenting, causes endothelial inflammation, thrombosis, and fibroproliferation. Systemic evidence of inflammation identifies patients at high risk of future coronary events, including those who appear to be healthy initially as well as those with stable and unstable coronary heart disease syndromes. Increases in serum C reactive protein (CRP) identify individuals at risk for future vascular events, including unstable angina, acute myocardial infarction, acute cerebrovascular accident, and sudden death. Similarly, systemic elevations in serum troponin 1, serum amyloid-like protein, fibrinogen, and interleukins-1, 2, 6, 8, and 18 identify patients with unstable angina and non-Q-wave myocardial infarction at increased risk for future coronary events. The presence of vascular inflammation may be detected by identifying temperature heterogeneity within plaques that demonstrate inflammation. In the future, the local evaluation of atherosclerotic plaques to detect the presence of inflammation coupled to measurements of systemic markers of inflammation, such as C-reactive protein, may help identify patients at increased risk and allow both local and systemic therapies that reduce their risk and prevent the development of acute coronary syndromes in at least some patients. PMID- 12077719 TI - Interventional therapy of the acute coronary syndromes. AB - The acute coronary syndromes (ACS) have in common rupture of a vulnerable plaque, leading to exposure of the subendothelial surface and plaque core. The resultant thrombosis leads to a variable degree of flow occlusion, the extent of which differentiates the three syndromes and their treatment by percutaneous coronary intervention (PCI). The guiding principle in the decision when to use PCI in the ACS is that the more time critical and high risk the clinical situation, the more likely it is that PCI will improve ultimate outcome. The use of risk stratification by clinical variables can lead to better triage of patients with non-ST-elevation myocardial infarction (MI) and unstable angina between PCI and medical management. Patients presenting with symptoms suggestive of prolonged ischemia should have an electrocardiogram searching for ST changes, a targeted physical, and blood drawn for rapid assay of cardiac enzymes. In the event that ST elevations suggest infarction, while medical therapy is initiated, emergency cardiac catheterization can be organized. PCI in ACS requires adjunctive antiplatelet and antithrombin therapy, and, in general, coronary stenting is advisable. Among patients with non-ST-elevation MI or unstable angina who can be medically stabilized, the presence of high clinical risk scores would favor early coronary angiography. In their absence, medical therapy can be pursued, unless recurrent ischemia occurs. When the patient's condition is stable, evaluation by stress testing can be used to guide further decisions. PMID- 12077721 TI - Pharmacogenomics in cardiovascular diseases. AB - Considerable heterogeneity exists in the way individuals respond to medications, in terms of both efficacy and safety. Inherited differences in the absorption, metabolism, excretion, and target for drug therapy have important effects on drug efficacy and safety. Pharmacogenomics aims to discover new therapeutic targets and understand genetic polymorphisms that determine the safety and efficacy of medications. The goal of pharmaco-genomics is customization of drug therapy with administration of a medication in an optimum dose that will be safe and effective with reduction in morbidity and mortality. PMID- 12077723 TI - Correlation of urine type I collagen-cross-linked N telopeptide levels with bone scintigraphic results in prostate cancer patients. AB - The diagnostic potential of a new bone resorption marker, type I collagen-cross linked N telopeptide (NTx), for bone metastasis of prostate cancer was evaluated. Ninty-one prostate cancer patients underwent bone scintigraphy, and urine NTx/creatinine (NTx/Cr) was measured. Urine NTx/Cr levels were compared with bone scintigraphic results. Urine NTx/Cr levels in the bone metastasis-positive group (n = 47) were 92.9 +/- 105.1 nmol/L of bone collagen, which is equivalent to per millimole of urinary creatinine (nmol/L BCE/mmol/L Cr), significantly higher than the level of the bone metastasis-negative group (n = 44) (59.0 +/- 41.6 nmol/L BCE/mmol/L Cr). When patients were classified by the extent of disease grade (EOD grade) nomenclature, the urine NTx/Cr level of the EOD (4+) group was 209.5 +/- 186.5 nmol/L BCE/mmol/L Cr. This level was significantly higher than those of the EOD (-) group (59.0 +/- 41.6 nmol/L BCE/mmol/L Cr), EOD (1+) group (59.0 +/- 47.8 nmol/L BCE/mmol/L Cr), and EOD (2+) group (81.1 +/- 41.3 nmol/L BCE/mmol/L Cr). However, no significant difference was observed between the EOD (-) and EOD (1+) groups. The mean change in urine NTx/Cr level 3 to 17 months after the first bone scintigraphy and urine NTx/Cr examination in the bone metastasis-progression group (n = 8) was 11.0 +/- 31.2 nmol/L BCE/mmol/L Cr, significantly higher than that in the bone metastasis-regression group (n = 15) (-26.8 +/- 40.7 nmol/L BCE/mmol/L Cr). In conclusion, urine NTx /Cr can be measured noninvasively and reflects the state of bone metastasis. However, the sensitivity of urine NTx/Cr is not as high as that of bone scintigraphy. Therefore, it may provide an auxiliary diagnostic index for bone scintigraphy. PMID- 12077722 TI - Effects of streptozotocin-induced diabetes on markers of skeletal muscle metabolism and monocarboxylate transporter 1 to monocarboxylate transporter 4 transporters. AB - Diabetes is known to alter both oxidative and glycolytic pathways in a fiber type dependent manner. In various skeletal muscles of normal rats, monocarboxylate transporter 1 (MCT1) has been found to be highly correlated to lactate uptake, as well as to oxidative capacity, whereas the distribution and characteristics of MCT4 make it a good candidate for the extrusion of lactic acid from glycolytic muscle cells. Since a previous study found decreased sarcolemmal lactate uptake in streptozotocin (STZ)-diabetic rats, we investigated the presence of MCT1 in relation to enzymatic markers of both oxidative and glycolytic pathways, as well as MCT4 content, in STZ-diabetic rats. Soleus (SOL), red tibialis anterior (RTA), extensor digitorus longus (EDL), heart, and preparations of purified sarcolemmal vesicles (SV) from control and STZ-diabetic rats were harvested for MCT1 and MCT4 content, citrate synthase activity (CS), and lactate dehydrogenase (LDH) isozymes. Basal blood lactate concentration was increased by 38% in the diabetic rats (close to 1.91 mmol/L). However, no change was found in either MCT1 or MCT4 content in these rats. The diabetic rats presented fiber type-specific decrease in CS activity. We noted a redistribution in LDH isozymes in diabetic muscles with a general increase in type H-LDH. Regression analyses indicated (1) a strong relationship between LDH-4 and LDH-5 and (2) MCT1 was still correlated with CS activity in diabetic muscles. These results suggest that diabetes-induced hyperlactatemia is not associated with changes in MCT1 or MCT4 expression, but with alterations of oxidative and glycolytic enzymes. PMID- 12077725 TI - Splanchnic galactose extraction is regulated by coingestion of glucose in humans. AB - When compared with galactose alone, coingestion of glucose with galactose decreases plasma galactose. The objective of this study was to determine if this was due to increased peripheral clearance or increased first pass clearance of galactose. Five adult volunteers were studied on 2 occasions during infusion of [6,6-(2)H(2)]glucose and [1-(13)C]galactose and ingestion of galactose alone at 11, 22, and 33 micromol x kg(-1) x min(-1) or galactose plus glucose at 11, 22, and 33 micromol x kg(-1) x min(-1) of each sugar. At 33 micromol x kg(-1) x min( 1) of galactose alone (1) plasma galactose increased to 2.3 +/- 0.3 mmol/L and galactose rates of appearance (Ra) to 18.3 +/- 1.6 micromol x kg(-1) x min(-1); (2) plasma glucose and glucose Ra were unaffected; (3) splanchnic extraction of galactose plateaued at approximately 15 micromol x kg(-1) x min(-1); and (4) galactose became the primary source of glucose Ra (75% +/- 9%). Coingestion of glucose and galactose at 33 micromol x kg(-1) x min(-1) each resulted in (1) decreased plasma galactose (0.3 +/- 0.1 mmol/L) and galactose Ra (6.4 +/- 1.8 micromol x kg(-1) x min(-1)); (2) increased plasma glucose and insulin; (3) doubling of splanchnic extraction of galactose; and (4) decreased contribution of galactose to glucose Ra (11% +/- 4%). We conclude that coingestion of glucose with galactose increases the splanchnic extraction, but decreases the conversion of galactose to glucose. PMID- 12077724 TI - Four-week low-glycemic index breakfast with a modest amount of soluble fibers in type 2 diabetic men. AB - Low-glycemic index diets are associated with a wide range of benefits when followed on a chronic basis. The chronic effects, however, of the substitution of 1 meal per day are not well known in diabetic subjects. Therefore, we aimed to evaluate whether the chronic use of a low-glycemic index breakfast (low-GIB) rich in low-GI carbohydrates and a modest amount of soluble fibers could have an effect on lipemia at a subsequent lunch, and improve glucose and lipid metabolism in men with type 2 diabetes. A total of 13 men with type 2 diabetes were randomly allocated in a double-blind cross-over design to a 4-week daily intake of a low GI versus a high-GI breakfast separated by a 15-day washout interval. The low-GI breakfast was composed of whole grain bread and muesli containing 3 g beta-glucan from oats. Low-GIB induced lower postprandial plasma glucose peaks than the high GIB at the beginning (baseline, P <.001) and after the 4-week intake (P <.001). The incremental area under the plasma glucose curve was also lower (P <.001, P <.01, baseline, and 4 weeks, respectively). There was no effect on fasting plasma glucose, insulin, fructosamine, or glycosylated hemoglobin (HbA(1c)). Fasting plasma cholesterol, as well as the incremental area under the cholesterol curve, were lower (P <.03, P <.02) after the 4-week low-GIB period than after the high GIB period. Apolipoprotein B (apo B) was also decreased by the 4-week low-GIB. There was no effect of the low-GI breakfast on triacylglycerol excursions or glucose and insulin responses at the second meal. The high-GIB, however, tended to decrease the amount of mRNA of leptin in abdominal adipose tissue, but had no effect on peroxisome proliferator-activated receptor gamma (PPARgamma) and cholesterylester transfer protein (CETP) mRNA amounts. In conclusion, the intake of a low-GI breakfast containing a modest amount (3 g) of beta-glucan for 4 weeks allowed good glycemic control and induced low plasma cholesterol levels in men with type 2 diabetes. The decrease in plasma cholesterol associated with low-GI breakfast intake may reduce the risk of developing cardiovascular complications in subjects with type 2 diabetes. PMID- 12077726 TI - Significance of beta2-adrenergic receptor gene polymorphism in obesity and type 2 diabetes mellitus in Korean subjects. AB - Catecholamines play a central role in the regulation of energy expenditure, in part stimulating lipid mobilization through lipolysis in fat cells. The beta(2) adrenergic receptor (ADRB2) is a major lipolytic receptor in human fat cells, and a recent study has shown that common polymorphisms occurring in codons 16 and 27 of the ADRB2 gene are significantly associated with obesity and lipolytic ADRB2 function in adipose tissue. We investigated whether previously described human ADRB2 gene polymorphisms are associated with obesity and diabetes in Korean subjects. According to the World Health Organization (WHO) criteria for oral glucose tolerance testing, 57 subjects had normal glucose tolerance (NGT), 32 had impaired glucose tolerance (IGT), and 106 had diabetes mellitus. The nondiabetic group (including NGT and IGT) consisted of 46 obese (defined as those with body mass index [BMI] of >or= 27 kg/m(2)) and 43 nonobese subjects (BMI < 27 kg/m(2)). The subjects with diabetes consisted of 62 obese and 44 nonobese subjects. There was no significant difference between nonobese and obese subjects in the allele frequency of ADRB2 gene polymorphisms at codons 16 and 27. There were no significant differences in BMI, percentage body fat, waist-to-hip ratio (WHR), systolic blood pressure, diastolic blood pressure and concentrations of fasting plasma glucose, fasting serum insulin, serum low-density lipoprotein (LDL) cholesterol, serum high-density lipoprotein (HDL)-cholesterol, serum triglyceride, and serum free fatty acids, according to ADRB2 gene polymorphisms at codons 16 and 27. The frequency of the Glu27 homozygote was 1.1%. These findings suggest that genetic variability in the ADRB2 gene may not be a major determinant for the development of obesity and diabetes in Koreans. PMID- 12077727 TI - Elevated serum ceruloplasmin levels in subjects with metabolic syndrome: a population-based study. AB - Serum ceruloplasmin was reported to be an independent risk factor for cardiovascular disease. We investigated whether serum ceruloplasmin level is elevated in subjects with metabolic syndrome (MS, insulin resistance syndrome) in a community-based population. A total 883 subjects over 40 years of age were studied among a population of the Chongup district, a rural area of South Korea. Serum ceruloplasmin levels were measured, and oral glucose tolerance tests were performed. Known cardiovascular risk factors, such as serum lipids, fasting insulin level, and urinary albumin excretion rate (UAER), were also measured. Serum ceruloplasmin levels in the subjects with MS (n = 167, 325 +/- 141 mg/L) were significantly higher than in those without MS (278 +/- 93 mg/L, P <.001). The mean ceruloplasmin level also increased as the glucose tolerance worsened (278 +/- 95 mg/L in normal glucose tolerance [NGT], 303 +/- 108 mg/L in impaired glucose regulation, and 328 +/- 148 mg/L in diabetes; P <.001). Serum ceruloplasmin level was positively correlated with age, fasting glucose, postload 2-hour glucose, total cholesterol, triglyceride, systolic blood pressure, diastolic blood pressure, and UAER and negatively with high-density lipoprotein (HDL)-cholesterol. In multiple regression analysis, serum ceruloplasmin level was independently associated with age, fasting glucose, triglyceride, HDL cholesterol, and UAER. In conclusion, serum ceruloplasmin level is elevated in the subjects with MS, as well as in subjects with impaired glucose regulation or diabetes mellitus. In addition, serum ceruloplasmin level is associated with various cardiovascular risk factors. These results suggest that elevated serum ceruloplasmin level can be a marker for metabolic stresses associated with MS. PMID- 12077728 TI - Metformin treatment lowers asymmetric dimethylarginine concentrations in patients with type 2 diabetes. AB - This study was initiated to see if plasma asymmetric dimethylarginine (ADMA) concentrations decreased in hyperglycemic patients with type 2 diabetes following metformin treatment, either as monotherapy or following its addition to sulfonylurea-treated patients. Fasting plasma glucose, dimethylarginine, and L arginine concentrations were measured before and 3 months after the administration of a maximally effective dose of metformin to 31 patients with type 2 diabetes in poor glycemic control (fasting plasma concentrations > 9.7 mmol/L), while being treated with either diet (n = 16) or a maximal amount of a sulfonylurea compound (n = 15). Fasting plasma glucose concentration (mean +/- SEM) decreased to a similar degree (P <.01) in patients treated with either metformin alone (12.4 +/- 0.5 to 9.5 +/- 0.5 mmol/L) or when it was added to a sulfonylurea compound (14.1 +/- 0.5 to 10.6 +/- 0.9 mmol/L). The improvement in glycemic control was associated with similar decreases (P <.01) in ADMA concentrations in metformin (1.65 +/- 0.21 to 1.18 +/- 0.13 micromol/L) and sulfonylurea + metformin-treated patients (1.75 +/- 0.13 to 1.19 +/- 0.08 micromol/L). Plasma L-arginine concentrations were similar in the 2 groups at baseline and did not change in response to metformin. Thus, metformin treatment was associated with a favorable increase in the plasma L-arginine/ADMA ratio. These results provide the first evidence that plasma ADMA concentrations decrease in association with improved glycemic control in patients with type 2 diabetes and demonstrate that the magnitude of the change in metformin-treated patients was similar, irrespective of whether it was used as monotherapy or in combination with sulfonylurea treatment. PMID- 12077729 TI - Intestinal rather than hepatic microsomal triglyceride transfer protein as a cause of postprandial dyslipidemia in diabetes. AB - Postprandial dyslipidemia may be a major cause of atherosclerosis in diabetes. Microsomal triglyceride transfer protein (MTP) is essential for the synthesis of the chylomicron particle in the intestine and very low-density lipoprotein (VLDL) in the liver. The purpose of the present study was to examine the effect of diabetes on MTP mRNA expression in a rabbit model of diabetes, which develops atherosclerosis. Male New Zealand white rabbits were fed a 0.5% cholesterol diet. Diabetes was induced with alloxan monohydrate. The lymphatic duct was cannulated and lymph collected for isolation of chylomicrons by ultracentrifugation. Apolipoprotein B48 (apo B48) and apo B100 were separated by polyacrylamide gradient gel electrophoresis and quantified by densitometry. MTP mRNA was determined in liver and intestine by RNase protection analysis, and MTP activity was measured. Diabetic animals had significantly increased plasma triglyceride and decreased high-density lipoprotein (HDL) cholesterol (P <.05). They also secreted more lymph chylomicron apo B48 and apo B100 (P <.05) and more lymph chylomicron total and esterified cholesterol/h (P <.05). Lymph chylomicron particles in the diabetic animals contained significantly less lipid/apo B (P <.05). Intestinal MTP activity and mRNA were significantly higher in diabetic compared with control rabbits (0.07 +/- 0.01 v 0.04 +/- 0.015 fluorescent units/microg microsomal protein and 66 +/- 21 v 37 +/- 11 amol MTP mRNA/microg total RNA (P <.005). There was no difference in MTP activity or mRNA expression in the liver. This study suggests that MTP may play an important role in the postprandial dyslipidemia of diabetes. PMID- 12077730 TI - Effects of apolipoprotein E genotype on dietary-induced changes in high-density lipoprotein cholesterol in obese postmenopausal women. AB - Lipid responses to a dietary intervention are highly variable between individuals. Part of this variation may be accounted for by individual differences in lipid-regulating genes that interact with diet to induce changes in lipoprotein metabolism. This study determined whether apolipoprotein E (APOE) genotype affects lipid responses to a low-fat, low-cholesterol diet in obese, postmenopausal women. Body weight and lipoprotein lipid responses to a 10-week, dietary intervention (American Heart Association [AHA] Step I) were compared in 61 women with the APOE 2/3 and APOE 3/3 genotype (APOE4-) and 18 women with the APOE 3/4 genotype (APOE4+) of a similar age, body composition, and maximal aerobic capacity. Body weight decreased by 2% in both groups, but changes in body weight correlated only with changes in low-density lipoprotein-cholesterol (LDL C) (r =.27, P <.05). The dietary intervention decreased total cholesterol and LDL C to a similar degree in both genotype groups. However, APOE4- women decreased high-density lipoprotein-cholesterol (HDL-C) by 17% +/- 11% and increased triglycerides by 20% +/- 41% in response to the diet, while APOE4+ women had a smaller decrease in HDL-C (-8% +/- 12%) and no change in plasma triglyceride. These group differences were significant for HDL-C (P <.01) and approached significance for triglycerides (P =.08). Moreover, APOE4- women decreased HDL(2) C by 32% +/- 45%, while APOE4+ women increased HDL(2)-C by 12% +/- 62% (P <.01 between groups). It may be prudent to genotype older women before initiating low fat diet therapy, as those with the APOE4 allele benefit the most, while the lipid profile could worsen in women without the APOE4 allele. PMID- 12077731 TI - Chronic leptin treatment normalizes basal glucose transport in a fiber type specific manner in high-fat-fed rats. AB - The aim of this investigation was to determine if a high-fat diet impaired and subsequent leptin administration improved non-insulin-stimulated (basal) glucose transport in rodents. Twenty-four male Sprague Dawley rats were divided into 1 of 2 groups: (1) normal diet (control [CON], n = 8) or (2) high-fat diet (n = 16) and received standard rat chow or a high-fat diet, respectively, for 12 weeks. The high-fat diet animals were then further subdivided into high fat (HF) (n = 8) or high-fat-leptin (HF-LEP) (n = 8) groups. The HF-LEP animals were injected with leptin (10 mg leptin/kg/d), while the CON and HF animals received vehicle over a 12-day treatment period. Following the 12-day treatment period, all animals were subjected to hind limb perfusion to assess rates of basal skeletal muscle 3-O methyl-D-glucose (3-MG) transport. Compared with the CON group, rates of 3-MG transport were reduced in the soleus (sol) and plantaris (plant) of the HF, but not the HF-LEP animals. Differences in skeletal muscle 3-MG transport could not be accounted for by an altered GLUT1 protein concentration. In contrast, a high fat diet reduced and chronic leptin treatment normalized the skeletal muscle GLUT4 protein concentration. The results indicate that a high-fat diet reduces and subsequent leptin treatment improves basal skeletal muscle glucose transport in a fiber-type-specific manner, but these changes do not appear to be due to alterations in the GLUT1 protein concentration. PMID- 12077732 TI - Body composition and hormonal responses to a carbohydrate-restricted diet. AB - The few studies that have examined body composition after a carbohydrate restricted diet have reported enhanced fat loss and preservation of lean body mass in obese individuals. The role of hormones in mediating this response is unclear. We examined the effects of a 6-week carbohydrate-restricted diet on total and regional body composition and the relationships with fasting hormone concentrations. Twelve healthy normal-weight men switched from their habitual diet (48% carbohydrate) to a carbohydrate-restricted diet (8% carbohydrate) for 6 weeks and 8 men served as controls, consuming their normal diet. Subjects were encouraged to consume adequate dietary energy to maintain body mass during the intervention. Total and regional body composition and fasting blood samples were assessed at weeks 0, 3, and 6 of the experimental period. Fat mass was significantly (P .2). The present study clearly demonstrates that an injection of intracellular Ca(2+) chelator in the HFD rats reverses insulin resistance, as well as normalizes elevated [Ca(2+)](i) in the insulin target cells. The results strongly support that sustained high levels of [Ca(2+)](i) in the insulin target cells may play an important role in insulin resistance, at least in the HFD rats. PMID- 12077742 TI - Effects of high- and low-isoflavone (phytoestrogen) soy foods on inflammatory biomarkers and proinflammatory cytokines in middle-aged men and women. AB - This study sought to determine effects of high- and low-isoflavone soy protein foods on acute-phase proteins and proinflammatory cytokines and whether isoflavone phytoestrogens might act as estrogens, which enhance the immune response. Forty-one hypercholesterolemic men and postmenopausal women underwent three 1-month diets consisting of a low-fat dairy food control phase and high- and low-isoflavone soy food test phases (50 g/d and 52g/d soy protein, respectively, and 73 mg/d and 10 mg/d isoflavone, respectively). Diets were low in saturated fat (<5% of energy) and cholesterol (<50 mg/d). Fasting blood analytes and blood pressure were measured at the start and end of each phase. For the entire group of subjects, no treatment differences were observed for acute phase proteins or proinflammatory cytokines. However, a significant interaction was noted between diet and sex. Assessing the results of men and women separately, women showed significantly higher interleukin-6 (IL-6) values after the high-isoflavone soy diet (P =.013) compared to control values. For women, the difference between the high- and low-isoflavone IL-6 values was significant using the unadjusted data (P =.048) but not after adjustment. No significant effects were seen for men or women in C-reactive protein (CRP), serum amyloid A (SAA), or tumor necrosis factor-alpha (TNF-alpha). Thus, high levels of isoflavone intake appear to increase serum concentrations of IL-6 in women. This finding may indicate an estrogenic effect of soy isoflavones in enhancing the immune response and provide a possible explanation through enhanced immune surveillance for lower incidence of certain cancers in soy-eating parts of the world. PMID- 12077743 TI - Immobilization stress alters intermediate metabolism and circulating lipoproteins in the rat. AB - In humans, stress can increase the risk of cardiovascular disease by altering lipoprotein metabolism. Scarce experimental and clinical data are available on this effect. Therefore, we studied the metabolic response to acute and chronic stress following a model of immobilization (IMO) in rats and we evaluated the resulting circulating lipoprotein levels. Repeated IMO treatment (2 hours daily, always between 9:00 AM and 11:00 AM, for 2 periods of 5 and 4 consecutive days, separated by 2 days of rest) daily decreased body weight gain and food intake, increased adrenal weight, and slightly reduced liver glycogen and plasma insulin (without considerable variations of blood glucose), which is characteristic of chronic stress. A single IMO application (30 minutes of an unexpected IMO starting at 2:00 PM immediately before the animals were killed) significantly increased the circulating levels of corticosterone, glucose, insulin, glycerol, and ketone bodies, which is the typical response to acute stress. Both acute and chronic stress decreased the plasmatic triacylglycerol (TAG) concentration, as reflected by the reduction in the number of very-low-density lipoprotein (VLDL) particles. This may be due to an increase in the metabolization of TAG, as suggested by the slightly higher amounts of circulating LDLs. Chronic stress, but not acute stress, significantly increased both the number and the estimated size of circulating high-density lipoprotein (HDLs), as shown by the plasma cholesterol concentration. Acute stress did not have an additive effect over chronic stress on the lipoprotein parameters studied. The metabolic effects of these IMO-induced alterations on lipoprotein profiles are discussed, and future studies in lipidic metabolism are suggested. PMID- 12077744 TI - High serum concentrations of soluble E-selectin correlate with obesity but not fat distribution in patients with type 2 diabetes mellitus. AB - Serum concentrations of soluble adhesion molecules, eg, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin are elevated in patients with type 2 diabetes. However, little is known about the role of obesity or abnormal fat distribution in inducing upregulation of adhesion molecules. To investigate this issue, soluble ICAM-1, VCAM-1, and E-selectin levels were evaluated in 40 obese and 30 nonobese patients with type 2 diabetes. Both groups were matched for age, sex, and glycosylated hemoglobin (HbA(1c)) levels. Computed tomography (CT) was used to measure the abdominal subcutaneous and visceral fat areas. Soluble ICAM-1 and VCAM-1 levels did not differ significantly between obese and nonobese patients. However, serum concentrations of soluble E-selectin were significantly higher in obese than in nonobese patients (90 +/- 7 v 56 +/- 4 ng/mL, P <.01). Soluble E-selectin levels significantly correlated with body mass index, subcutaneous fat area, and visceral fat area (Rho = 0.48, 0.37, and 0.30, respectively). Stepwise multiple regression analysis showed that body mass index (F = 16.7), but not subcutaneous and visceral fat areas (F = 0.29 and 0.01, respectively), significantly and independently correlated with soluble E-selectin levels. Our results suggest that obesity may induce endothelial activation or increased shedding of cell surface E selectin that leads to subsequent increase in soluble E-selectin levels. The high serum concentrations of E-selectin closely correlated with increased total fat volume, but not with regional fat distribution. PMID- 12077745 TI - Decline in muscle mass with age in women: a longitudinal study using an indirect measure. AB - Muscle mass is known to decline with age, but only limited longitudinal data exist to quantify the rate of loss. Using 24-hour urine creatinine, corrected for the contribution of dietary meat intake, we assessed the change with age prospectively in 107 women who provided a minimum of 3 sets of measurements spanning an average interval of 11.9 years, centered around age 55 years. The rate of change in 24-hour urine creatinine at that age averaged -0.94%/yr (95% confidence interval [CI], -1.24% to -0.64%/yr; P <.001). Change in creatinine excretion was directly correlated with change in weight (P <.01), with those gaining weight tending to gain both lean and fat mass. The rate of change in creatinine excretion with age in our subjects is similar to that described in published cross-sectional studies on age-related change in total body potassium and in longitudinal studies using (40)K and dual-energy xray absorptiometry. Our study is the largest longitudinal study reported to date and provides, perhaps, a more secure basis for estimating muscle mass change with age than has been available heretofore. PMID- 12077747 TI - Stephen L. Gans overseas lecture. Mass screening for neuroblastoma in Japan: lessons learned and future directions. AB - BACKGROUND/PURPOSE: Since 1985, a nationwide mass screening program (MS) for neuroblastoma has been conducted for 6-month-old infants throughout Japan, resulting in the detection of more than 1,900 cases of neuroblastoma. The outcome of these patients has been excellent: more than 97% of them are alive. Yet, several reports suggest that the number of advanced-stage neuroblastoma patients over 1 year of age has not changed substantially. The current report focuses on the 15-year experience with MS of the Kyushu Pediatric Oncology Study Group. METHODS: The clinical and biological features of neuroblastoms detected (n = 320) and not detected by MS (n = 245) were compared. Regional and national statistics for neuroblastoma before and after 1985 were analyzed using standard epidemiologic measures for the occurrence of disease. RESULTS: The majority of the MS-positive cases were biologically favorable and had an excellent outcome. In contrast, the majority of non-MS patients in whom neuroblastoma later developed had advanced-stage, unfavorable-prognosis tumors. The overall mortality rate of neuroblastoma in the Kyushu area was not improved by MS. CONCLUSIONS: The optimal time for screening is the point at which neuroblastomas regressing spontaneously can no longer be detected, but more aggressive disease can be found. A birth cohort study could determine the optimal timing for a second screening. Identification of other new prognostic factors may be required. PMID- 12077748 TI - Vaginal reconstruction for ambiguous genitalia and congenital absence of the vagina: a 27-year experience. AB - BACKGROUND/PURPOSE: Gender assignment to a neonate with ambiguous genitalia is crucial. Patients with an absent vagina require the construction of an artificial vagina. In an effort to improve care, the authors have categorized their experience with this group of children. METHODS: Since 1974, we cared for 114 patients with anomalies of the genitalia. There were 53 genotypic girls with congenital adrenal hyperplasia (CAH), 16 genotypic boys with testicular feminization syndrome (TFS), 13 with mixed gonadal dysgenesis (MGD), 9 with Mayer Rokitansky syndrome, and 4 true hermaphrodites. The remaining 19 had other genital abnormalities. RESULTS: After 1980, patients with CAH underwent clitoral recession and vaginoplasty. All patients with TFS were raised as girls and underwent orchidectomy. Eleven of the MGD patients were given a female sex assignment and underwent gonadectomy. Twenty-eight patients underwent intestinal vaginoplasty including 8 of the TFS patients, 9 with Mayer-Rokitansky syndrome, 8 patients with cloacal anomalies, 2 patients for rhabdomyosarcoma, and 1 of the MGD patients. CONCLUSIONS: (1) This review emphasizes the range of diagnoses the surgeon must be prepared to address in patients with ambiguous genitalia. (2) Colovaginoplasty is an excellent procedure for replacement of a completely absent vagina. (3) Continued evaluation of this group will delineate appropriate timing and choice of procedure. PMID- 12077749 TI - Rectovaginal fistula: a common diagnostic error with significant consequences in girls with anorectal malformations. AB - BACKGROUND/PURPOSE: Rectovaginal fistula is a term that continues to be used frequently to describe girls with anorectal malformations. This study attempts to evaluate the true frequency of this anomaly and the consequences of its misdiagnosis. METHODS: A retrospective review of all girls with anorectal malformations treated by the senior author from 1980 through September 2000 was performed, and the pertinent literature was reviewed. RESULTS: Of the 617 patients identified, only 6 were found to have a true rectovaginal fistula, an incidence of 1%. A total of 139 of the 617 patients were referred after a previous repair. Of these, 42 had a diagnosis of recto-vaginal fistula originally. The diagnosis was incorrect in all 42. Twelve patients had a rectovestibular fistula, and 30 had a cloaca. The 30 cloaca patients, because of the misconception that they had a rectovaginal fistula, underwent an initial repair of only the rectal component of the malformation, leaving all patients with a urogenital sinus necessitating a second complete repair. CONCLUSIONS: The use of the term rectovaginal fistula is common, despite a true incidence of 1% in girls with anorectal malformations. The majority of girls will have either a rectovestibular fistula or a cloaca. Such diagnostic errors may lead to the use of inappropriate surgical techniques, incomplete repair, and unnecessary morbidity. PMID- 12077750 TI - Routine MRI evaluation of low imperforate anus reveals unexpected high incidence of tethered spinal cord. AB - BACKGROUND/PURPOSE: Previous clinical practice has included evaluation for the presence of tethered cord in those children who have imperforate anus with a high lesion. To define the incidence in children with low lesions, the authors reviewed their experience with a protocol employing routine magnetic resonance imaging (MRI), regardless of the level of the lesion, to determine the presence of a tethered cord in all children with imperforate anus. METHODS: A retrospective review of children with imperforate anus was conducted over the last 13 years at our institution. Lesions were categorized as high versus low based on the supralevator or infralevator position of the fistula. RESULTS: Sixty three patients completed evaluation for a tethered cord. Twenty-two (34.9%) of these 63 patients had a tethered cord: 11 of 41 (26.8%) patients with high lesions and 11 of 22 (50.0%) of those with low lesions. Of those children with a low lesion, 83% of the boys had a tethered cord, whereas 38% of the girls had a tethered cord. Forty-five percent of the patients with low lesions and a tethered cord did not have any other lumbosacral anomalies. All 22 children with a tethered cord underwent surgical release. CONCLUSIONS: The incidence of tethered cord in children with low lesions of imperforate anus is not lower than those with high lesions. The authors advocate early evaluation of all children with imperforate anus for a tethered cord. PMID- 12077751 TI - Successful thoracoscopic lung biopsy in children utilizing preoperative CT-guided localization. AB - BACKGROUND/PURPOSE: Although thoracoscopy in children is a well-accepted technique, there are limitations to this approach. Small lung nodules or those not located on the pleural surface often require an open thoracotomy. The purpose of this report is to evaluate the utility of computed tomography (CT) guided needle localization of lung nodules in children followed by thoracoscopic resection. METHODS: From 1997 to 2000, 13 thoracoscopic procedures were performed on 12 children who presented with small pulmonary nodules (<1 cm) or nodules deep to the pleural surface. Preoperative needle localization was performed by injecting methylene blue under CT guidance. Patients then were taken directly to the operating room for thoracoscopic resection. RESULTS: All 13 procedures were completed successfully thoracoscopically. Twelve yielded diagnostic pathologic findings as well as therapeutic resections in 7. All patients underwent extubation in the operating room, and chest tubes were removed within 36 hours. No thoracotomies were performed, and there were no complications. CONCLUSIONS: Preoperative localization utilizing CT guidance is safe and beneficial in children with pulmonary nodules less than 1 cm in size and those located deep to the pleural surface. This technique allows the surgeon to perform thoracoscopy for diagnosis and therapeutic resection of these lesions rather than submitting patients to the morbidity of a thoracotomy. PMID- 12077752 TI - Thoracoscopic placement of phrenic nerve electrodes for diaphragmatic pacing in children. AB - BACKGROUND/PURPOSE: Diaphragmatic pacing can provide chronic ventilatory support for children who suffer from congenital central hypoventilation syndrome (CCHS) or cervical spinal cord injury. The authors report a new thoracoscopic approach for establishing diaphragm pacing. METHODS: Between 1997 and 2000, 9 children ranging in age from 5 to 15 years and suffering from these disorders underwent thoracoscopic placement of bilateral phrenic nerve electrodes. A 3- or 4-trocar technique was used to dissect the phrenic nerve in the midchest and suture a phrenic nerve electrode (Avery Laboratories I-110A, Commack, NY) into place. The electrode was tunneled to a subcutaneous pocket in the upper abdomen and attached to an implanted pacing unit. RESULTS: Bilateral electrodes were placed successfully into all patients. The average procedure time was 3.3 hours (range, 2.5 to 4.6), and average hospital stay was 4.2 days (range, 3 to 5). Four patients experienced postoperative complications (pneumonia, atelectasis, bradycardia, and pneumothorax). Average follow-up has been 30 months (range, 15 to 49). Eight patients have reached their long-term pacing goals. CONCLUSIONS: Phrenic nerve electrodes can be implanted thoracoscopically and allow the successful use of diaphragmatic pacing therapy. Avoidance of thoracotomy with its associated perioperative morbidity and scarring may encourage wider utilization of diaphragmatic pacing in children. PMID- 12077753 TI - Experience with laparoscopic adrenalectomy in pediatric patients. AB - BACKGROUND: Laparoscopic adrenalectomy (LA) is now being recognized as the standard in the management of benign adrenal pathology in adult patients. Few reports have described the use of this technique in pediatric patients. This study combines experience from 2 institutions with lateral transperitoneal LA in children to analyze our results and the clinical and biochemical response to laparoscopic adrenalectomy in patients with hormonally active adrenal tumors. METHODS: A bi-institutional retrospective review of all patients undergoing LA between January 1997 and January 2001 was performed. Clinical and biochemical data were obtained during routine follow-up. RESULTS: Seventeen laparoscopic adrenalectomies were performed during this period. The average operating time was 120 minutes, mean estimated blood loss was 25 mL, the mean size of the adrenal lesion was 4.8 cm, and the mean length of hospitalization was 35 hours. Resolution of clinical and biochemical parameters of adrenal hyperfunction was accomplished in all patients with adrenocortical hyperplasia and pheochromocytoma in postoperative follow-up. CONCLUSIONS: Laparoscopic adrenalectomy can be performed safely and effectively with a short hospital stay, minimal blood loss, and excellent functional outcome in this age group. The authors believe laparoscopic adrenalectomy is an excellent approach for the management of benign pediatric adrenal pathology. PMID- 12077754 TI - Robotic-assisted esophagoesophagostomy. AB - BACKGROUND/PURPOSE: Minimally invasive repair of esophageal atresia has been described but remains technically challenging. Robotic surgical systems address many of these technical challenges. The purpose of this study was to develop the procedure for and evaluate the technical feasibility of performing a robotic assisted esophagoesophagostomy using the Zeus Robotic Surgical System. METHODS: Esophagoesophagostomy was performed in 10 piglets using thoracoscopic (control, n = 5) and robotic-assisted (Zeus, experimental, n = 5) approaches. An interrupted esophageal anastomosis using intracorporeal knot tying techniques was performed and evaluated for leak, narrowing, caliber, and mucosal approximation. Anesthesia, operative, anastomotic, and robotic set-up times were recorded as was the number of stitches used. RESULTS: All 10 anastomoses were patent with no narrowing and with excellent mucosal approximation. One anastomosis in the control group had a small leak. There was no statistically significant difference between the groups for the parameters measured. Weight (kg): control (C), 6.4 +/- 0.8; experimental (E), 6.3 +/- 1.0, P =.08. Times (min): anesthesia, C-124 +/- 25, E-151 +/- 20, P =.09; operative, C-97 +/- 21, E-131 +/- 27, P =.06; anastomotic, C-89 +/- 20, E-125 +/- 34, P =.08; robotic set-up, C-6.4 +/- 9.3, E 15.6 +/- 20, P = 0.13. Stitches (No.): C-11.8 +/- 0.8, E-12.0 +/- 1.2, P =.7. Caliber (French):C-18F-5; E-18F-4, 14F-1. CONCLUSION: Robotic-assisted esophagoesophagostomy is technically feasible and offers an alternative approach to thoracoscopic repair of esophageal atresia. PMID- 12077755 TI - Central hepatic resection for pediatric tumors. AB - BACKGROUND/PURPOSE: Resection of hepatic malignancies in childhood has been facilitated greatly by an understanding of hepatic segmental anatomy. This report documents the indications and technique of central hepatic resection (mesohepatectomy) in children with hepatic malignancies. METHODS: This is a retrospective study, and data are obtained from patient charts and operative reports. Three patients underwent central hepatic resection on or between June 27, 1997 and October 1, 1999. RESULTS: All patients were boys, and their ages at diagnosis were 0.3, 0.8, and 3.8 years. Two had hepatoblastoma and one central hepatic metastases from a poorly defined epithelial malignancy. All 3 received neoadjuvant chemotherapy before hepatic resection. Segments IV, V, and VIII were removed in 2, and IV and V in the third. All patients survived, and histopathologic margins were clear in each. One patient had a postoperative bile collection treated with percutaneous catheter drainage. At follow-up, all patients remain alive without evidence of disease, and all have normal hepatic function. CONCLUSION: Central hepatic resection of malignant tumors involving segments IV, V, and VIII is feasible and effective in childhood. PMID- 12077756 TI - Development of an internet-based protocol to facilitate randomized clinical trials in pediatric surgery. AB - BACKGROUND/PURPOSE: Randomized clinical trials (RCTs) perhaps are the best method available for the comparison of therapeutic interventions, yet these studies remain scarce in the pediatric surgery literature. The infrequency of surgical conditions, parents' refusal to participate, and difficulty with standardizing operative technique are potential obstacles to conducting multicenter randomized trials. Novel applications of information technology can potentially overcome these obstacles and facilitate randomized trials in pediatric surgery. METHODS: Through collaboration with our Biostatistics Department, an Internet-based protocol was developed to facilitate our own multicenter trial for the surgical treatment of necrotizing enterocolitis. A secure, on-line data entry system was developed to manage all aspects of patient care data. Data are validated during the entry phase and directly uploaded into a clinical database for safety monitoring and analysis. Multimedia resources available online were developed to encourage standardization of surgical and nonoperative therapy. Comprehensive informed consent, educational and other support resources are available on line for the parents and family members of patients. These include an 8-minute video detailing pathophysiology, treatment options, and important informed consent issues surrounding the trial. The website also provides administrative support to study centers and periodic updates on trial progress to the medical community at large. CONCLUSIONS: Internet-based protocols may facilitate trials in pediatric surgery by reducing administrative costs, improving standardization of surgical technique, and potentially increasing enrollment success rates. Future investigation will examine the impact of this protocol on the education, knowledge base, and attitudes of parents, as well as determine the efficiency and overall cost effectiveness of this data management strategy. PMID- 12077757 TI - The placenta as a cell source in fetal tissue engineering. AB - PURPOSE: This study was aimed at determining whether fetal tissue constructs can be engineered from cells derived from the placenta. METHODS: A subpopulation of morphologically distinct cells was isolated mechanically from specimens of human placenta (n = 6) and selectively expanded. The lineage of these cells was determined by immunofluorescent staining against multiple intermediate filaments and surface antigens. Cell proliferation rates were determined by oxidation assays and compared with those of immunocytochemically identical cells derived from human amniotic fluid samples (n = 6). Statistical analysis was by analysis of variance (ANOVA). After expansion, the cells were seeded onto a polyglycolic acid polymer/poly-4-hydroxybutyrate scaffold. The resulting construct was analyzed by both optical and scanning electron microscopy. RESULTS: The immunocytochemical profile of expanded placental cells was consistent with a nontrophoblastic, mesenchymal origin. Their proliferation rate in culture was not significantly different when compared with mesenchymal fetal cells isolated from human amniotic fluid; however, it was greater than previously reported rates for similar cells obtained from postnatal or adult tissues. Construct analysis showed dense layers of cells firmly attached to the scaffold without evidence of cell death. CONCLUSIONS: Subpopulations of nontrophoblastic, mesenchymal cells can be isolated consistently from the human placenta. These cells proliferate as rapidly as fetal mesenchymal amniocytes in vitro and attach firmly to polyglycolic acid scaffolds. The placenta can be a valuable and practical source of cells for the engineering of select fetal tissue constructs. PMID- 12077758 TI - Fetal tissue engineering: in utero tracheal augmentation in an ovine model. AB - BACKGROUND/PURPOSE: This study was aimed at comparing fetal tissue engineering with autologous free grafting in an ovine model of in utero tracheal repair. METHODS: Chondrocytes were isolated from both elastic and hyaline cartilage specimens harvested from fetal lambs and expanded in vitro. Cells were seeded dynamically onto biodegradable scaffolds, which then were maintained in a rotating bioreactor for 6 to 8 weeks. Constructs subsequently were implanted into fetal tracheas (n = 15), in a heterologous fashion (group I). In group II, fetuses (n = 5) received autologous free grafts of elastic cartilage harvested from the ear as tracheal implants. In vivo specimens were harvested for histologic analysis at different time-points postimplantation. RESULTS: In the 12 of 15 surviving fetuses of group I, all constructs were found to resemble normal hyaline cartilage, engraft well despite their heterologous origin, and display time-dependent epithelialization derived from the native trachea. All autologous free grafts were engrafted and epithelialized at birth, retaining histologic characteristics of elastic cartilage, but were more deformed than engineered constructs. Of the lambs allowed to reach term, 5 of 5 in the engineered group and 4 of 5 in the free graft group could breathe spontaneously. CONCLUSIONS: (1) Tissue-engineered cartilage, as well as autologous free grafts, can be implanted successfully into the fetal trachea, resulting in engraftment and function. (2) Engineered cartilage provides enhanced structural support after implantation into the fetal trachea when compared with free grafts. Prenatal tracheoplasty may prove useful for the treatment of severe congenital tracheal malformations. PMID- 12077759 TI - Positive intrapulmonary oncotic pressure enhances short-term lung growth acceleration after fetal tracheal occlusion. AB - PURPOSE: This study was aimed at determining whether positive oncotic pressure induced in the fetal lung liquid could safely maximize accelerated lung growth after tracheal occlusion. METHODS: Fetal lambs (n = 21) were divided into 4 groups: group I (n = 5) consisted of sham-operated controls; group II (n = 5) underwent simple tracheal occlusion (TO); group III (n = 5) received TO and 60 mL of saline injected into the trachea; and group IV (n = 6) underwent TO and intratracheal infusion of 60 mL of iso-osmotic, 6% Dextran 70. All fetuses were delivered near term, at a mean of 15.9 +/- 1 days postoperatively. Their lungs were studied by standard morphometric techniques, and the basic chemical profile of the lung liquid was analyzed. Statistical comparisons were by 1-way analysis of variance (ANOVA) and post-hoc analyses by the Bonferroni correction for multiple comparisons, with P values less than.05 considered significant. RESULTS: The lung volume-to-body-weight ratio (LV:BW) was significantly different among groups. Pairwise comparisons of LV:BW showed that it was higher in group IV than in all other groups, but there was no difference between groups II and III. Airspace fraction was not significantly different among groups, and histologic appearance was normal in all lung samples. There were no differences in lung liquid osmolarity, pH level, and electrolyte concentrations. CONCLUSION: Positive intrapulmonary oncotic pressure by an isosmotic agent boosts short-term lung growth acceleration after fetal tracheal occlusion with no evidence of cell damage. PMID- 12077760 TI - Enteral feeding tubes are a reservoir for nosocomial antibiotic-resistant pathogens. AB - BACKGROUND: Patients and their surroundings are known reservoirs for nosocomial pathogens. Enteral feeding tubes and formula are not thought of as reservoirs for nosocomial organisms. METHODS: A prospective observation study was conducted comparing methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant Enterococcus (VRE) cultured from nosocomial infections and MRSA/VRE cultured from enteral feeding tubes used in the same neonatal intensive care unit during the same time period but in different babies. DNA fingerprinting then was used to compare MRSA and VRE cultured from feeding tubes with MRSA/VRE isolates cultured from clinical infections. RESULTS: There were 23 S aureus isolates; 12 of 23 were methicillin resistant (MRSA). There were 4 MRSA infections in patients without feeding tubes. DNA fingerprinting showed that the MRSA species causing each of the clinical infections also was found in the feeding tubes of other babies. There were no vancomycin-resistant Enterococcus infections during the study period. CONCLUSION: Feeding tubes are a reservoir for antibioticresistant pathogens that can be transmitted to other infants. PMID- 12077761 TI - Principles for the surgical management of patients with Proteus syndrome and patients with overgrowth not meeting Proteus criteria. AB - BACKGROUND: Proteus syndrome is a rare, sporadic disorder consisting of disproportionate overgrowth of multiple tissues, vascular malformations, and connective tissue or epidermal nevi. Patients with Proteus syndrome present with diverse and variable phenotypes because of the syndrome's mosaic pattern of distribution. METHODS: Eighty patients with Proteus syndrome, satisfying published diagnostic criteria, and 51 patients with overgrowth not meeting Proteus criteria were identified from the literature. Three additional patients, one patient with Proteus syndrome and 2 patients with overgrowth, were treated at the author's institutions and are discussed in detail. All nonorthopedic and noncutaneous surgical interventions were reviewed. RESULTS: Fourteen genitourinary, 9 gastrointestinal, and 5 otolaryngologic operations were performed on patients with Proteus syndrome. Six genitourinary, 5 gastrointestinal, and 2 otolaryngologic operations were performed on patients with overgrowth not meeting Proteus criteria. Eight patients with Proteus syndrome and 4 patients with overgrowth experienced thoracic manifestations, generally diffuse cystic pulmonary lesions, but only 1 of 12 underwent surgical treatment. CONCLUSIONS: Patients with visceral manifestations of either Proteus syndrome or overgrowth not meeting Proteus criteria should be treated in a similar manner. Lesions involving the ovaries and testes, because of the high incidence of neoplasm, should be managed aggressively. Gastrointestinal and renal lesions may be managed conservatively with frequent follow-up to minimize abdominal explorations. All patients undergoing surgery should have a thorough preoperative assessment of their airway and pulmonary reserve because of the relatively high frequency of tonsillar hypertrophy and pulmonary cystic involvement. PMID- 12077762 TI - Laparoscopic Nissen fundoplication for reactive airway disease. AB - BACKGROUND: Surgical management for gastroesophageal reflux disease (GERD) induced reactive airway disease in children has been shown to be superior to medical therapy. Laparoscopic Nissen fundoplication is a safe and effective procedure in children. METHODS: The authors performed a retrospective review of 24 patients who underwent a laparoscopic Nissen fundoplication for documented GERD and reactive airway disease. RESULTS: Persistent cough was the primary symptom in 22 of 24 patients, and all but one had lipid laden macrophages on bronchoscopy. The mean length of hospital stay was 2.7 days. There were no major postoperative complications. Eighteen of 24 patients are symptom free and off all medications an average of 17 months postoperatively. The average medication burden of the 6 remaining patients was reduced from 6.8 to 2.3 medications. CONCLUSIONS: Children with reactive airway disease who do not respond to medical therapy should undergo a workup for GERD. These preliminary results suggest that laparoscopic Nissen fundoplication is a potentially effective treatment for pulmonary manifestations of GERD. PMID- 12077763 TI - Management of congenital esophageal stenosis. AB - BACKGROUND/PURPOSE: The authors report the incidence, diagnosis, and treatment methods of congenital esophageal stenosis (CES) at their institution. METHODS: A retrospective analysis of 123 patients with tracheoesophageal anomalies in a pediatric hospital between 1980 and 1999 was performed. Charts were reviewed for patient demographics, presence of true CES, associated congenital anomalies, method of diagnosis, methods of treatment, and histopathology. RESULTS: Six of the 123 patients (4.9%) had true CES (one patient had 2 separate stenoses). Two patients had isolated CES, one patient had CES with isolated tracheoesophageal fistula (TEF), one patient had CES with isolated esophageal atresia (EA), and 2 patients had CES with EA/TEF. Diagnoses were made with an upper gastrointestinal (GI) contrast study in 5 patients, and one patient had one of 2 stenoses diagnosed by prenatal ultrasound and the other diagnosed intraoperatively. Four of the 7 stenoses were treated with surgical resection, and the remainder was treated with esophageal dilatation. Histopathology from the 4 resected stenoses showed tracheobronchial remnants in 3 specimens and submucosal thickening in 1 specimen. CONCLUSIONS: Although isolated CES is rare (2 of 123 = 1.6%), CES associated with other tracheoesophageal anomalies has a higher incidence (4 of 123 = 3.25%). Patients with this lesion should be treated first with dilatation. If ineffective, resection is required. PMID- 12077764 TI - Surgical management of open versus laparoscopic adrenalectomy: outcome analysis. AB - PURPOSE: The authors sought to compare the outcome of children undergoing open versus laparoscopic adrenalectomy for an adrenal tumor. METHODS: Medical records of children that underwent an adrenalectomy from 1990 through 1999 were reviewed. Sixty-four adrenalectomies were performed: 27 pheochromocytomas, 36 neuroblastomas, and 1 virilizing tumor. Sixty adrenalectomies were performed open and 4 laparoscopically. The patient's age, surgical length of stay, operative charge, hospital cost, operating time, blood loss, and outcome were examined. RESULTS: Mean age for an open procedure was 8.9 +/- 0.9 years and 14 +/- 1.1 for laparoscopic (P =.019). Surgical length of stay for open was 5.4 +/-.38 days and 2.7 +/-.62 days for laparoscopic (P =.006). Patient operative charges were $12,941 +/- 676 for laparoscopic and $4,714 +/- 411 for open (P <.001). When total estimated patient cost, including hospital stay, were compared between groups there was no significant difference. Similar mean operating times and blood loss were noted. There were no deaths or complications in children with a pheochromocytoma. The mortality rate in children with neuroblastoma was 28%. CONCLUSIONS: Adrenalectomy for benign tumors can be performed safely. In selected children a laparoscopic procedure can be expected to decrease the surgical length of stay without increasing operating time or complications. PMID- 12077765 TI - Clinical evaluation of serum levels of matrix metalloproteinases and tissue inhibitors of metalloproteinases as predictors of progressive fibrosis in postoperative biliary atresia patients. AB - BACKGROUND/PURPOSE: The authors analyzed serum Metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in postoperative biliary atresia (POBA) patients to evaluate their usefulness for assessing progressive fibrosis. METHODS: The authors measured serum MMPs (-1,2) and TIMPs(-1,2) using respective one-step sandwich enzyme immunoassays in 36 POBA patients (mean age, 12.3 +/- 6.5 years) and 13 age-matched controls. POBA patients were classified into 3 groups; group I: n = 12; normal liver function test (LFT); group II: n = 15; elevated LFT, anicteric; and group III: n = 9; elevated LFT, icteric. RESULTS: Serum MMP-2 levels were higher in all BA groups(especially in group II and III) compared with controls (P <.01). However, MMP-1 in all POBA patients groups (especially in group II and III) was significantly decreased compared with controls (P <.01). There was no statistical difference between mean values for serum TIMP-1 and TIMP 2 in all POBA patients compared with controls. CONCLUSIONS: Serum MMPs levels might be useful for predicting the progress of fibrosis and the functional reserve in POBA patients with documented deterioration in liver function caused by fibrosis. Our finding that TIMPs do not rise as cirrhosis progresses and this imbalance may be a causal factor for the excessive accumulation of extracellular matrix, which, in turn, may contribute to the progression of fibrosis in BA. PMID- 12077766 TI - Engineering a composite neotrachea with surgical adhesives. AB - BACKGROUND/PURPOSE: Reconstructive surgery often is limited by the availability of normal tissue. Tissue engineering provides promise in the development of "artificial tissues." The purpose of this study was to test the efficacy and viability of the use of a biologic surgical adhesive TISSEEL in combining engineered bronchial epithelium with engineered cartilage. METHODS: Using isolated human cells, bronchial epithelium and mature cartilage were engineered. Using a contact adhesive technique, TISSEEL was used to biologically fuse the bronchial epithelium and the cartilage. The fused composite then was supported for 5 days in tissue culture. The mechanical properties of the adhesion were tested, and the construct was studied morphologically to assess viability of the cartilage and the bronchial epithelium. The bronchial epithelium showed a normal cell size (337.2 microm2) and epithelial thickness (46.47 microm). RESULTS: TISSEEL was effective in fusing the epithelium to the cartilage. The construct remained viable for 5 days in culture. There was no difference in the dimensions of the bronchial epithelium or the epithelial cells. Mechanical adhesion was achieved. CONCLUSIONS: Biologically compatible fibrin glue is an effective surgical adhesive that allows the tissue types to be fused while remaining viable and morphologically accurate. Surgical adhesives may show promise in the development of composite tissue development in the field of bioengineering. PMID- 12077767 TI - Levels of circulating antiinflammatory cytokine interleukin-1 receptor antagonist and proinflammatory cytokines at different stages of biliary atresia. AB - BACKGROUND/PURPOSE: Antiinflammatory cytokine (interleukin [IL]-1ra) and proinflammatory cytokines (tumor necrosis factor [TNF]- alpha and IL-6) appear to play important roles in the pathophysiology of liver disease. In this study, the authors attempt to identify the roles of IL-1ra, IL-6, and TNF-alpha at different stages in postoperative biliary atresia (POBA) patients. METHODS: Serum concentrations of IL-1ra, IL-6, and TNF-alpha were measured in 34 POBA patients and 25 healthy controls using specific enzyme-linked immunosorbent assays (ELISA). POBA patients were classified into 5 groups: group I (n = 21), less than 3 years old; Ia, stable; Ib, mild liver dysfunction; Ic, severe liver dysfunction. Group II (n = 13), (more than 3 years old); IIa, stable; IIb, variable liver dysfunction. RESULTS: For IL-1ra differences between controls and groups Ib, Ic, and IIa were statistically significant (P <.01), and for IL-6, differences between controls and groups Ia, Ib, Ic, and IIb were statistically significant (P <.01). Groups Ib and Ic (less than 3 years old subjects) had the highest levels in this series. Serum TNF-alpha was not detected in any groups. CONCLUSION: The relationship between serum levels of IL-6 and IL-1ra may be predictive of prognosis with respect to the progression of fibrosis in POBA patients. PMID- 12077768 TI - Altered neutrophil function in the neonate protects against sepsis-induced lung injury. AB - BACKGROUND/PURPOSE: Neutrophils (PMNs) are well known effectors of lung injury after sepsis. The accumulation of PMNs into the lung is dependent on a complex cascade of events that includes the local production of chemokines. Interestingly, neonates are protected from lung injury after zymosan-induced sepsis. The authors hypothesized that this protection was caused by either altered PMN function or diminished lung chemokine production compared with the adult. METHODS: Sepsis was induced in neonatal and adult rats by an intraperitoneal injection of zymosan. Animals were killed 24 hours later and lungs examined for PMN accumulation and function, chemokine production, and lung injury. RESULTS: Septic neonates (SN) were protected from pulmonary edema when compared with septic adults (SA). Lung PMN number and chemokine (MIP-2) production increased in both septic neonates and adults when compared with vehicle (V) treated animals. Conversely, PMN function was decreased significantly in neonates when compared with adults. CONCLUSIONS: Despite equivalent lung PMN accumulation and chemotactic protein production, PMN function and lung injury in septic neonates was diminished when compared with that of adults. These findings suggest that neonates may be relatively protected from sepsis-induced lung injury caused by immature PMN function. PMID- 12077769 TI - Long-term follow-up of patients with necrotizing enterocolitis. AB - BACKGROUND/PURPOSE: Despite the rise in the incidence of necrotizing enterocolitis (NEC), there is a paucity of data regarding long-term patient outcome. The authors examined functional outcome of infants with NEC (n = 103) treated at our institution between 1991 and 1995. METHODS: The authors reviewed the medical records of infants who were treated both operatively and nonoperatively, n = 103. Variables examined included gestational age, birth weight, Bell stage (I through III), operations performed, and mortality rate. Telephone interviews assessed school enrollment, developmental delay, bowel function, and nutritional status. RESULTS: Children treated operatively had a lower gestational age than those in the nonoperative group. Likewise, birth weight in the operative group was significantly lower. Sixty-three percent of patients had stage III, and the remainder had stage II disease. The telephone response rate was 61%. Mean age at follow-up was 7.5 +/- 2.5 years. All children ate by mouth. Nearly all children were toilet trained. All children were less than the 50th percentile for height and weight, and the majority (83%) were enrolled in school full time. CONCLUSION: Infants with stage II and III NEC who are treated operatively or nonoperatively have a favorable long-term outcome. PMID- 12077770 TI - Adeno-associated virus (AAV)-mediated gene transfer in respiratory epithelium and submucosal gland cells in human fetal tracheal organ culture. AB - BACKGROUND/PURPOSE: Since the discovery of the cystic fibrosis transmembrane regulator (CFTR) gene, cystic fibrosis has been an attractive target for gene therapy. Postnatal gene transfer in the respiratory epithelium has been difficult and particularly inefficient in the submucosal gland cells, the target cells for CFTR gene transfer. The authors hypothesized that during development, there is a favorable environment for fetal gene therapy with fewer physical barriers to efficient gene transfer and more accessible epithelial and submucosal gland precursor cells. The authors tested this hypothesis in a novel human fetal tracheal organ culture system using a serotype 2 recombinant AAV that contains an enhanced green fluorescent protein (eGFP) reporter gene (AAV-CMV-eGFP). METHODS: Human fetal tracheas at between 16 and 20 weeks' gestation age were used in this study. The distal end of each trachea was ligated and secured in an upright position with the open proximal end facing up. Only the ante-lumenal surface was exposed to culture media. 5 x 10(9) particles of AAV-CMV-eGFP were administered intratracheally through the open end. Fetal tracheas were maintained in tracheal organ culture media and harvested at either 2 weeks (n = 3) or 4 weeks (n = 7) after injection. The fetal tracheas were processed for routine H&E, standard electron microscopy (EM), and fluorescence microscopy for analysis of eGFP transgene expression. RESULTS: Histology confirmed the preservation of structural integrity out to 4 weeks of fetal tracheal organ culture. EM showed intact tight junctions of the apical respiratory epithelium. At 2 weeks after AAV-CMV-eGFP injection, there was minimal transgene expression. However, at 4 weeks there was extensive transgene expression in not only the respiratory surface epithelium but also the submucosal gland cells of the human fetal tracheal organ culture. Transgene expression was seen in nearly all cells in the submucosal glands. CONCLUSIONS: AAV-mediated gene transfer in human fetal tracheal organ culture was highly efficient with excellent transgene expression at 4 weeks in both respiratory surface epithelium and submucosal gland cells. This highly efficient gene transfer may result from fewer physical barriers and more accessible target precursor cells. These results are consistent with more efficient gene transfer in fetal tracheobronchial epithelium and show the potential for fetal gene therapy using AAV for the treatment of congenital airway disease such as cystic fibrosis. PMID- 12077771 TI - Engraftment of bone marrow and fetal liver cells after in utero transplantation in MDX mice. AB - BACKGROUND/PURPOSE: In utero hematopoietic stem cell transplantation (IUHSCTx) has been experimentally or clinically effective only in circumstances in which there is a survival advantage for donor cells. A survival advantage exists for normal muscle cells in muscular dystrophy. Because hematopoietic and mesenchymal stem cells may have the capacity to differentiate into muscle cells, the authors hypothesized that in utero bone marrow (BM) or fetal liver (FL) stem cell transplantation may be used to treat muscular dystrophy. METHODS: Time-dated 14 day-gestation fetal muscular dystrophy mice (mdx) were injected intraperitoneally with 1 to 5 x 10(6) BM or FL cells per fetus from Rosa26 donor mice (transgenic for lacZ). Four weeks after birth, peripheral blood from the pups was analyzed for hematopoietic chimerism by using fluorescence-activated cell sorting (FACS) for the Ly-9.1 marker. Chimeric mice (6 BM and 2 FL recipients) were sacrificed at 12 to 14 months of age, muscles were stained with X-gal, and analyzed by 1- to 2-microm plastic sections. Polymerase chain reaction (PCR) for lacZ was performed in other organs to determine systemic engraftment. RESULTS: At the time of death, all animals that were chimeric at 4 weeks continued to show hematopoietic chimerism of 0.2% to 9% by FACS. Engrafted donor cells were found in multiple sections from hindlimb skeletal muscles, diaphragms, and hearts from both BM and FL recipients. These cells had incorporated into the host muscles, and their morphology was consistent with myogenic differentiation. PCR of BM, liver, spleen, thymus, kidney, and lung for lacZ was positive in multiple animals. CONCLUSIONS: IUHSCTx leads to widespread engraftment of donor cells in multiple muscle compartments of hematopoietic chimeras. The advantage for normal myocytes offered in the mdx model allows engraftment and myogenic differentiation of transplanted BM or FL cells by morphology at a relatively higher frequency in muscle relative to other tissues, without the need for host conditioning. Because muscular dystrophy now can be detected early in gestation, such a strategy may offer a future alternative in the clinical treatment of this disease. PMID- 12077773 TI - A comparison of laparoscopic and open pyloromyotomy at a teaching hospital. AB - BACKGROUND/PURPOSE: An increasing number of pediatric surgeons are using the laparoscopic approach to treat pyloric stenosis. The advantage of laparoscopic pyloromyotomy is uncertain and has not been evaluated in the setting of a pediatric surgery fellowship program. METHODS: The authors retrospectively reviewed the medical records of all patients who underwent pyloromyotomy for congenital hypertrophic pyloric stenosis at their institution from January 1, 1997 through December 31, 2000 (n = 117). Information obtained included age, sex, weight, admission laboratory values, attending surgeon, resident surgeon and their level of training, operating time, intraoperative and postoperative complications, time to full feedings, incidence of postoperative emesis, duration of postoperative emesis, length of stay, and total hospital charges. These variables then were compared between the open (OPEN) and laparoscopic (LAP) groups. RESULTS: From January 1, 1997 through December 31, 2000, 65 LAP and 52 OPEN pyloromyotomies were performed. Characteristics of patients in the OPEN and LAP groups were similar. The mean operating time was 33 +/- 2 minutes for OPEN versus 38 +/- 2 minutes for LAP (P =.07). The incidence of postoperative emesis (LAP, 68%, OPEN, 65%), duration of postoperative emesis (LAP, 7.3 +/- 1.2 hours; OPEN, 8.1 +/- 1.8 hours), and time to full feedings (LAP, 19.5 +/- 1.6 hours; OPEN, 19.5 +/- 1.3 hours) did not differ significantly between groups (P >.05). Mean postoperative length of stay in both groups was similar (LAP, 31 +/- 5; OPEN, 28 +/- 2 hours; P =.64). Mucosal perforation occurred in 5 patients (8%) in the Lap and 2 patients (4%) in the OPEN group (P =.39). Postoperative complications occurred in 12 LAP (18%) and 6 OPEN patients (12%, P =.31). Five LAP cases were converted to OPEN. In the LAP group there was one unrecognized mucosal perforation and one incomplete pyloromyotomy both of which required reoperation. As the laparoscopic approach was adopted, general surgery resident participation as operating surgeon in these cases decreased from 81% in 1997 to 19% in 2000. Hospital charges were higher in the LAP group, but not significantly (LAP, $6,676 +/- 1,005; OPEN, $5,292 +/- 306; P = 27). CONCLUSIONS: Laparoscopic pyloromyotomy has progressively become the dominant surgical approach to pyloromyotomy at our institution. The LAP and OPEN approaches have similar outcomes. However, the Lap approach may be associated with increased complication rates, a reduction in general surgery resident operative experience, and higher hospital charges. PMID- 12077772 TI - Thyroid transcription factor-1 expression in the human neonatal tracheoesophageal fistula. AB - BACKGROUND/PURPOSE: Esophageal atresia with tracheoesophageal fistula (EA/TEF) is a relatively common congenital anomaly, but its pathogenesis remains poorly understood. Previous studies using the experimental Adriamycin-induced rat model of EA/TEF suggest that the fistula tract, or "distal esophagus," is derived from respiratory epithelium and expresses the respiratory-specific transcription factor TTF-1. To better correlate the experimental rat model of EA/TEF with the human anomaly, we looked for evidence of a respiratory-derived origin in the neonatal TEF. METHODS: After IRB approval, 2 human TEF samples were removed at the time of surgery. The tissue from the fistula tract was trimmed in accordance with what the surgeons deemed to be appropriate for the preparation for a primary anastomosis. The tissues then were processed for reverse transcription polymerase chain reaction (RT-PCR), histology, and immunohistochemistry. Normal embryonic lung cDNA was used for positive controls. RESULTS: Histologic examination of tissue specimens showed many epithelial tubules within loose connective tissue and a disorganized muscular coat. Thyroid transcription factor one (TTF-1) and hepatocyte nuclear factor 3beta (HNF-3beta) were shown to be present in the tissue specimen by RT-PCR and immunohistochemistry. In addition, FGF-10, a strong morphogen present in the developing lung, and FGF-1 were both present by RT-PCR. The PCR band sizes for both FGF-1 and -10 were appropriate, using human embryonic cDNA as a control, and the bands were confirmed as nongenomic by either placing the PCR primers across a known intron sequence (TTF-1) or by absence of a band in a negative RT control (HNF-3beta, FGF-10, FGF-1). CONCLUSIONS: The presence of TTF-1 in the neonatal TEF shows, for the first time, that parallels may be drawn between the experimental rat model of TEF and the human anomaly at the molecular level. Moreover, these results suggest that the fistula tract in the human neonate is derived from a respiratory cell lineage. This respiratory origin of the human TEF may explain the poor esophageal motility (and subsequent serious respiratory complications) of the distal segment after standard EA/TEF repair. PMID- 12077774 TI - Streamlining the care of patients with hypertrophic pyloric stenosis: application of a clinical pathway. AB - PURPOSE: The purpose of this study was to evaluate the impact of a clinical pathway on infants admitted to a pediatric tertiary care center with the diagnosis of hypertrophic pyloric stenosis (HPS). METHODS: The records of 132 HPS patients were evaluated before and after implementation of a clinical pathway. Infants were excluded for prematurity, admission to nonsurgical services, or multiple diagnoses requiring prolonged hospitalization, resulting in 83 patients for analysis. Group I (prepathway, n = 40) and group II (postpathway, n = 43) infants were analyzed for time from admission to operation, operation to first feeding, operation to discharge, total length of stay, hospital charges, metabolic status at time of admission, and postoperative complications. The Mann Whitney test was performed (statistical significance at P <.05). RESULTS: There was no significant difference between group I and group II patients in the length of preoperative hospitalization or metabolic status at the time of hospital admission. In comparison with group I patients, there was a significant reduction in time to resumption of oral feedings (4.6 +/- 1.9 hours v 7.5 +/- 3.2 hours; P <.001) for group II infants and a significantly earlier discharge (26.7 +/- 6.8 hours v 38.0 +/- 11.7 hours; P <.001). This resulted in a shortened length of stay (41.8 +/- 9.7 hours v 57.8 +/- 14.3 hours; P <.001) with an associated decrease in hospital charges ($4,555 +/- $464 v $5,400 +/- $1,017; P <.001). CONCLUSIONS: Elimination of practice variability by the use of a clinical pathway for HPS resulted in significant reduction of hospital stay and related charges. The impact of the pathway occurred in the postoperative period and is a consequence of a rapid and systematic return to oral feedings. PMID- 12077775 TI - Transfection of the sodium/glucose cotransporter into colon mucosa: a novel treatment for short bowel syndrome. AB - BACKGROUND/PURPOSE: Short bowel syndrome results from small intestine loss but frequently is associated with survival of the colon. This study was designed to determine if colonic mucosa could be induced to absorb galactose by tranfection of the sodium glucose cotransporter, SGLT-1 into a colonic segment. METHODS: Using 10 rats, a 7-cm segment of colon was infused for 1 hour with a solution containing 50 microg/mL of a plasmid with or without an SGLT-1 insert. An 80% small bowel resection was performed, and the segment was interposed into the small bowel. On the third day [14C] galactose absorption was measured. Mucosal RNA was extracted, and relative band intensities were measured using primers for SGLT-1. Statistical analysis was performed using the Student's t test and expressed as mean +/- SEM. RESULTS: Rats transfected with the SGLT-1 plasmid showed a significant increase (194%) in galactose absorption compared with controls. Transfected animals also showed high levels of of SGLT-1 transcription when compared with controls (792% increase). CONCLUSIONS: These data show that in vivo exposure of colon mucosa to a plasmid containing SGLT-1 allows transfer of that gene into enterocytes. Expression of SGLT-1 can create an absorptive segment that may in part alleviate the malabsorption associated with short bowel syndrome. PMID- 12077776 TI - Heparin-binding EGF-like growth factor preserves crypt cell proliferation and decreases bacterial translocation after intestinal ischemia/reperfusion injury. AB - BACKGROUND/PURPOSE: Heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF), a known mitogenic, chemotactic, and cytoprotective growth factor for epithelial cells, was examined to see whether it could protect intestinal barrier function and decrease bacterial translocation (BT) after ischemia/reperfusion (I/R) injury. METHODS: In vitro, tight junctional integrity of intestinal epithelial cells (IEC-6) cells was evaluated by measuring transepithelial electric resistance (TEER), and monolayer permeability was evaluated by translocation of Escherichia coli C25. In vivo, crypt cell proliferation was assessed by 5-bromodeoxyuridine incorporation with calculation of a proliferative index (PI), and BT was evaluated by culture of mesenteric lymph nodes. RESULTS: In vitro, anoxia damaged tight junctional integrity and increased permeability of IEC-6 cell monolayers, events that were reversed completely by treatment of the cells with HB-EGF. Twenty-four hours after I/R injury in vivo, crypt cell proliferation index (PI) decreased significantly from 35.6 +/- 4.5 to 17.8 +/- 3.4. Administration of HB-EGF preserved crypt cell activity with PI of 34.9 +/- 4.1, similar to that of normal ileum. None of the normal or sham-operated animals showed BT, whereas BT occurred in 87.5% of I/R injured rats. In animals exposed to I/R but treated with HB-EGF, BT was decreased significantly to 12.5%. CONCLUSION: HB-EGF preserves proliferation of crypt cells, maintains integrity of epithelial cells, and subsequently decreases enteric BT after I/R injury. PMID- 12077777 TI - Laparoscopic duodenoduodenostomy for duodenal obstruction in infants and children. AB - BACKGROUND/PURPOSE: Duodenal obstruction, such as that resulting from atresia or web, routinely has been corrected by laparotomy and duodenoduodenostomy. Until recently, no one has reported on the use of minimally invasive techniques to correct this congenital anomaly. Over the last 6 months we have approached 4 patients, 3 with atresia and one with a web, laparoscopically. Three were newborns, and one was 8 months old. METHODS: All procedures were performed with 3 mm instruments and scopes. RESULTS: Operating time in all cases was less then 90 minutes. Visualization was excellent, and there were no intraoperative complications. Feedings were started on postoperative day 5 in all 3 neonates and day 3 in the infant. All 4 were on full feedings after 3 days. Follow-up upper gastrointestinal tests show no evidence of stricture or obstruction. CONCLUSION: Laparoscopy provides an excellent way to evaluate and treat congenital duodenal obstruction. PMID- 12077778 TI - Minimally invasive bar repair for 'redo' correction of pectus excavatum. AB - BACKGROUND/PURPOSE: A small percentage of patients who have undergone traditional, "Ravitch-type" pectus excavatum repair present with unsatisfactory results and require a second procedure for correction. Reoperative open surgery for pectus excavatum has been associated with extensive dissection and substantial blood loss. The minimally invasive (MIS) bar repair for the correction of pectus excavatum has been gaining acceptance. This study evaluates the authors results with patients who have undergone the MIS bar repair for redo correction of their pectus excavatum. METHODS: A retrospective chart review of all patients undergoing MIS bar repair between December 1997 and August 2001 was performed. Information about demographics, deformity, operative course, complications, and early outcome was recorded. RESULTS: Ninety-two patients underwent MIS repair during this period. Ten patients had redo MIS bar repair for unsatisfactory prior open correction. Operating time was 52 minutes for standard patients and 70 minutes for the redo patients (P <.001). Blood loss and postoperative hospitalization were similar between groups. CONCLUSION: The minimally invasive pectus repair can be performed safely with minimal blood loss and short operating time in patients who have undergone prior unsatisfactory open repair of pectus excavatum and can be an alternative approach to reoperative open repair in these patients. PMID- 12077779 TI - Hepatocyte growth factor pretreatment reduces apoptosis and mucosal damage after intestinal ischemia-reperfusion. AB - BACKGROUND/PURPOSE: Ischemia-reperfusion (IR) injury to the intestine can result in mucosal damage and cellular death. This study was designed to evaluate the protective effects of pretreatment with hepatocyte growth factor (HGF) on intestine after moderate IR injury. METHODS: Control animals (n = 7) received 48 hours of intravenous saline, and treatment animals (n = 7) received HGF (150 microg/kg/d). After 35 minutes of mesenteric artery occlusion and 120 minutes of reperfusion, serum and jejunal mucosa samples were obtained. Fluorometric assays were performed for hexosaminidase A (HEX A) and beta-glucuronidase (GLUC), enzyme markers of enterocyte necrosis. Apoptosis was quantified by the TUNEL method. Transcription of tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) was assessed by multiplex reverse transcription polymerase chain reaction (RT-PCR) Statistical analysis was performed using the Student's t test. RESULTS: After HGF pretreatment, HEX A and GLUC activities were reduced from 543 +/- 28 to 343 +/- 35 nmole/h/mL (P <.01) and 183 +/- 29 to 119 +/- 22 nmole/h/mL (P <.01), respectively. Pretreated animals had a reduced number of apoptotic cells per 10 crypts (33 +/- 11) compared with untreated rats (225 +/- 24) after IR injury (P <.01). Mean IFN-gamma band intensity was lower in HGF-pretreated animals (0.05 +/- 0.02) compared with controls (0.31 +/- 0.09; P <.05). CONCLUSIONS: Pretreatment with HGF reduces the severe crypt apoptosis and cellular necrosis after IR injury to the intestine. These data suggest that HGF may be beneficial in attenuating IR damage and thus may have significant clinical application. PMID- 12077780 TI - Development of a model for prediction of survival in pediatric trauma patients: comparison of artificial neural networks and logistic regression. AB - BACKGROUND/PURPOSE: There is a paucity of outcome prediction models for injured children. Using the National Pediatric Trauma Registry (NPTR), the authors developed an artificial neural network (ANN) to predict pediatric trauma death and compared it with logistic regression (LR). METHODS: Patients in the NPTR from 1996 through 1999 were included. Models were generated using LR and ANN. A data search engine was used to generate the ANN with the best fit for the data. Input variables included anatomic and physiologic characteristics. There was a single output variable: probability of death. Assessment of the models was for both discrimination (ROC area under the curve) and calibration (Lemeshow-Hosmer C Statistic). RESULTS: There were 35,385 patients. The average age was 8.1 +/- 5.1 years, and there were 1,047 deaths (3.0%). Both modeling systems gave excellent discrimination (ROC A(z): LR = 0.964, ANN = 0.961). However, LR had only fair calibration, whereas the ANN model had excellent calibration (L/H C stat: LR = 36, ANN = 10.5). CONCLUSIONS: The authors were able to develop an ANN model for the prediction of pediatric trauma death, which yielded excellent discrimination and calibration exceeding that of logistic regression. This model can be used by trauma centers to benchmark their performance in treating the pediatric trauma population. PMID- 12077781 TI - Posterior urethral valves. PMID- 12077782 TI - Meconium dependence of bowel damage in gastroschisis. PMID- 12077783 TI - Bowel plication in addition to primary anastomosis in patients with jejunal atresia. PMID- 12077784 TI - Infants undergoing hypospadias repair the stent must be kept with the proximal end in the bladder. PMID- 12077785 TI - RV dialysis: a dream realized and a dream threatened. Part II. PMID- 12077786 TI - [E-mail--a blessing or a curse?]. PMID- 12077787 TI - [White and red. A story of a different pro-Finnish man]. PMID- 12077788 TI - [The question of paternity throughout the times]. PMID- 12077789 TI - [Surgery in candle light: the illumination of operating rooms in the 19th Century]. PMID- 12077790 TI - [Ciguatera--a tropical fish poisoning. 400 hundred years of history and the present situation]. PMID- 12077791 TI - [Peaceful Christmas, Santa Claus]. PMID- 12077792 TI - [The old people's truth and that of the scientific society's--about fish tapeworm infection and a little bit about its therapy]. PMID- 12077793 TI - [What is expensive and what is cheap?]. PMID- 12077794 TI - [A religious experience or a psychiatric symptom?]. PMID- 12077795 TI - [Is a big head an academic burden--observations on the head circumference of honorary doctors at Helsinki University]. PMID- 12077796 TI - [Does the choice of specialty have influence on how many children the doctor will have in his/her own family?]. PMID- 12077797 TI - [Lady doctors who specialize in gynecology drink from paper cups]. PMID- 12077798 TI - [Does the drinking of mineral water help to control the weight?]. PMID- 12077799 TI - Developing your creativity. PMID- 12077800 TI - The Internet and electronic mail. PMID- 12077801 TI - Cutaneous manifestations of HIV and HCV. AB - Human immunodefiency virus (HIV) and hepatitis C virus (HCV) infections have had major effects on the field of medicine over the past 2 decades. Dermatologic manifestations are common and may be the presenting complaint in either disease. Clinicians should be aware of these cutaneous conditions so that the underlying disease may be properly diagnosed. PMID- 12077802 TI - Dermatological manifestations of hepatitis C. AB - Hepatitis C (HCV) infection is a leading cause of chronic liver disease in the United States. Certain dermatologic conditions are associated with HCV infection. Awareness of these conditions will facilitate better evaluation and management of patients by health care providers. Nurses can also increase awareness and understanding of HCV infection through patient education. These efforts may encourage earlier assessment and intervention. PMID- 12077803 TI - What's your assessment? Neonatal acne. PMID- 12077804 TI - The rising incidence of melanoma in children and adolescents. AB - Although melanoma is a rare occurrence in childhood and adolescence, recent reports indicate a rise in its incidence. This concerning trend is largely attributed to increasing amounts of sun exposure. Health care providers can intervene by the early identification and treatment of melanomas and educating children about sun-protective behaviors. PMID- 12077805 TI - Wound assessment and evaluation. PMID- 12077806 TI - Latex allergy: creating a safe environment. AB - Latex allergy is a serious problem for health care providers and the general public. Understanding the problem and following several simple suggestions can safeguard patients and workers in health care settings. PMID- 12077807 TI - Factors influencing consumers' selection of a primary care physician. AB - There is a growing body of literature regarding patient choice of health care plans, patient satisfaction, and patient evaluation of health care quality, but there is little information concerning the factors that influence the initial selection of a primary care physician (PCP). This exploratory study identifies and conceptualizes the physician selection dimensions which include: physician reputation/manner, physician record, physician search, consumer self-awareness, physician location, physician qualifications, physician demographics, office atmospherics, house calls/insurance, and valuing patient opinion. The study also develops and tests a scale for PCP selection using factor analysis which is demonstrated to be valid, and determines significant differences of variables, which include education level, gender, and age, using a summated scale. The study is of use to physicians in their targeting and communication strategies, and to researchers seeking to refine the scale. PMID- 12077808 TI - Understanding the role consumer involvement plays in the effectiveness of hospital advertising. AB - Both intensified competition and greater consumer participation in the choice process for healthcare has increased the importance of advertising for health care providers and seriously challenged many of the preconceptions regarding advertising. This study investigates the effectiveness of advertising under conditions of high and low involvement using the Elaboration Likelihood Model to develop hypotheses that are tested in a 2 x 2 x 2 experimental design. The study findings provide insights into the influence of message content and message source on consumers categorized as high or low involvement. It was found that consumers classified as high-involvement are more influenced by a core service relevant message than those consumers classified as low-involvement. Moreover, a non-physician spokesperson was found to have as much or more influence as a physician spokesperson regardless of the consumers' involvement level. PMID- 12077809 TI - The U.S. infant formula industry: is direct-to-consumer advertising unethical or inevitable? AB - Throughout their history, U.S. based infant formula companies have promoted their products as though they required a prescription. This form of marketing is called "ethical" promotion, which focuses on gaining a physician to parent recommendation for a brand of infant formula. Until Nestle's entry into the U.S. infant formula market in 1988, there was little direct-to-consumer promotion of infant formula. This article provides a background on the history of infant formula practices in the United States and then focuses on a descriptive model to explain how mothers' make their infant formula selection. Finally, we offer suggestions for the "ethical" marketers of infant formula. PMID- 12077810 TI - Selling open access health care delivery to patients and administrators: what's the hook? AB - A new concept in health care delivery involves the use of Open Access Scheduling for patients. In an attempt to manage spiraling medical costs and patient care demands many medical practices and managed care organizations are looking for alternative delivery models for health care. Open Access Scheduling has been garnering many advocates and converts from past traditional medical service delivery models. Unfortunately, due to its limited penetration into the medical community, little of Open Access' essential characteristics are generally understood. This paper looks at Open Access from the perspectives of: patients, medical administrators, office staff and providers. We discuss the tenets of Open Access, the benefits from its use, its challenges, and the steps necessary to initiate this type of service delivery. PMID- 12077811 TI - Essay: a cancer patient's perspective on how and why care providers should pursue healing partnerships. AB - This essay is based on the author's experiences as a cancer patient who underwent the grueling treatment known as a bone marrow transplant. An unintended consequence of the author's war with cancer is that he now "enjoys" more empathy for and insights into the challenging service encounters experienced by acutely ill individuals. This essay discusses why cancer patients and health service providers should each benefit when healing partnerships are developed. It also describes how such healing partnerships should be pursued. The content delivered in this essay is intended to help health care professionals better manage the means by which they deliver health services to acutely ill patients. PMID- 12077813 TI - [A new method of radiofrequency needle ablation of the prostate in the treatment of benign hyperplasia]. AB - The authors present the results of a new transvesical approach for radiofrequency needle ablation of the prostate in 29 patients with benign prostatic hyperplasia (BPH). The ablation has been done using a new radiofrequency generator ESHFG-440 40-1 made in Russia. Upon 12-month follow-up urination normalized in 23 patients. This result is better than that after transurethral needle ablation. The transvesical needle ablation has no contradictions and can be used in a wide range of patients with BPH both in hospital and outpatient settings. PMID- 12077812 TI - [Results of the effectiveness of long-term treatment of patients with benign prostatic hyperplasia]. AB - A long-term proskar (finasterid, MSD) treatment was given to 428 patients with benign prostatic hyperplasia (BPH). The response was achieved in 93.8-95.8% patients. The duration of the course should be at least 12 months. Side effects were rare: libido, erection, ejaculate volume diminished in 5.1, 5.6 and 4.2% patients, respectively. 20 patients were operated on after proskar therapy. Such pretreatment reduced glandular prostatic tissue. PMID- 12077815 TI - [Immunophenotyping of peripheral blood lymphocytes in patients with various forms of urethritis]. PMID- 12077814 TI - [Short-term outcomes of radical surgery for bladder cancer]. AB - A total of 102 radical cystectomies have been conducted for 1996-2001 (94 males, 8 females, age 37-78 years, mean age 57.5 years). Most of the patients (81.4%) were diagnosed to have transitional cell carcinoma. Supravesical urine derivation was made by means of ureterocutaneostomy and transureteroureteronephrostomy in 7(6.9) patients, ureterosigmoanastomosis--in 41(40.2%), Sigma-rectumpouch--in 1(0.9%) patients, ileocystoplasty, largely by Hautmann and Studer was carried out in 52(51%) patients. Postoperative complications developed in 25(24.5%) patients who often had adhesive ileus. Four patients died: 2 of pulmonary artery thromboembolism, 1 of acute cardiac failure and 1 of sepsis. Methods of continence urine derivation were preferred, such as ureterosigmoanastomosis by Mainz-Pouch II and creation of orthotopic bladder of the ileum. PMID- 12077816 TI - [Informative value of immunologic analysis of blood and ejaculate in diagnosing chronic prostatitis]. AB - Ejaculate and blood serum from 40 patients with chronic prostatitis were studied immunologically, IgG, IgA, sIgA, IgM concentrations in ejaculate may be greatly influenced by multicomponent composition of ejaculate. This means that it is impossible to diagnose chronic prostatitis only by changes of immunoglobulin concentrations in ejaculate. Data on IgA, sIgA concentrations plus on leukocyte count in ejaculate give more information for precise diagnosis of chronic prostatitis. PMID- 12077817 TI - [Use of sparfloxacin (Sparflo) in treating complicated urologic infections]. AB - The authors review the effectiveness of a novel fluoroquinolone drug sparfloxacine (Sparflo, Dr. Reddy's Laboratories) in the treatment of patients with complicated forms of pyelonephritis and prostatitis. Both gramnegative and grampositive agents causing complicated urologic infections were highly sensitive to sparfloxacine: Enterobacter spp.--40.8%, Pseudomonas aeruginosa--38.5%, Proteus spp.--42.6%, E. coli--91.4%, Staphylococcus spp.--80.0%, Enterococcus faecalis--21.4%. Sparfloxacine was used in the treatment of 43 patients with complicated pyelonephritis and prostatitis. The complicating factors were the following: urolithiasis, renal anomalies, hydronephrotic transformation, nephroptosis, benign prostatic hyperplasia. Clinical response reached 83.7%; microbiological--78.7%. Thus, sparfloxacin has a wide spectrum of antibacterial activity and can be effectively used in patients with complicated infections of the urinary tract. PMID- 12077818 TI - [A "small" kidney tumor]. AB - The authors think it necessary to distinguish small tumors of the kidney--a neoplasm 4 cm in size maximum. The number of such small tumors account for 1/3 of all renal newgrowths. An important role in detection of small tumors belongs to ultrasonography. Ultrasonic finding of the tumor should be followed by spiral computer tomography with 3-dimensional reconstruction of the image of the tumor, calyceal-pelvic system and renal vessels. Small tumors are asymptomatic in 71.4 cases, in many cases benign, in renal cell carcinoma their cells are well differentiated, they do not invade paranephral fat. Morphological verification is effective and safe in puncture biopsy of the tumor under ultrasound control. Adequate treatment depends on correct diagnosis of the tumor nature. Surgery of choice in small tumor of the kidney is resection. PMID- 12077819 TI - [Diagnosis and treatment of upper urinary tract infection in women as a postoperative complication for treating genital endometriosis]. AB - 156 women operated on for genital endometriosis received treatment for infection of the upper urinary tracts diagnosed by ultrasound. The treatment was conducted only in case of normal urodynamics achieved by drainage (on demand) and comprised antibacterial therapy, efferent hemocorrection, intravascular laser irradiation of blood, hyperbaric oxigenation. A response was obtained in 98% patients. PMID- 12077820 TI - [Effect of sodium hypochlorite on neutrophilic granulocytes during treatment for purulent-inflammatory diseases of retroperitoneal organs]. AB - The effect of systemic immunostimulation with autoblood oxidized with sodium hypochlorite is demonstrated in 79 patients with pyoinflammatory diseases of the retroperitoneal organs. Mechanism of phagocytic reaction to introduction of oxidized autoblood, which consists in extracorporeal activation of lymphocytes and monocytes from peripheral blood with their return to the donor, is outlined. The changes in phagocytic activity of neutrophilic granulocytes indicated that combined treatment of the patients provokes significant shifts in phagocytic activity of the cells: stage I phagocytosis accelerates and becomes less intensive, stage II is characterized by a high activity and in the majority of cases phagocytosis stage III is registered. PMID- 12077821 TI - [Polychemotherapy combined with low-intensity laser therapy in treating patients with renal tuberculosis]. AB - A prospective study of 150 patients with verified nephrotuberculosis has demonstrated feasibility of essential expansion of the therapeutic limit and of 1/3 reduction in the number of operations due to adjuvant radiation with low intensity laser beam used in a complex of etiopathogenetic treatment of the urinary system. PMID- 12077822 TI - [Role of physico-chemical and biochemical composition of urine in the genesis of combined nephrolithiasis and cholelithiasis]. AB - Enzymuria, principal fractions of phospholipids, crystal-inhibiting urine activity were measured in 120 patients with nephrolithiasis combined with cholelithiasis. The patients were divided into two groups: group 1-80 patients with symptoms of nephrolithiasis and latent cholelithiasis; group 2-40 patients with symptoms of cholelithiasis and latent nephrolithiasis. It is shown that hyperenzymuria/microproteinuria, high excretion of phospholipids and cholesterol with urine in patients with nephrolithiasis and its complications not only affect physico-chemical properties of urine promoting atypical salt crystallization with stone formation but also determine characteristics of phasic urine conditions. High urine concentration of phospholipid (acyl derivatives) degradation products and cholesterol in patients with combination of nephrolithiasis with cholelithiasis changes solubilizing and micello-forming ability of urine and can directly stimulate crystal formation and development of aseptic inflammation in renal parenchyma. Low total crystal-inhibiting urine activity, altered surface free energy due to surface-active substances, high calcium ionization and low urine magnesium ionization are not only pathognomonic criteria of nephrolithiasis but also parameters reflecting high risk of oxalate-calcium formation in patients with nephrolithiasis combination with cholelithiasis. PMID- 12077823 TI - [Extravesicle ureterocystoanastomosis]. AB - Extravesical ureterocystoanastomosis without wide opening of the urinary bladder was practised since 1986. A total of 79 patients were operated: 71 children at the age from 1 to 14 years and 8 adults aged 14 to 58 years. Surgical intervention was performed on 112 ureters for vesicoureteral reflux and supravesical ureteral obstruction. 6 of 79(7.6%) patients on postoperative day 11 15 after removal of intubating drains had an attack of acute pyelonephritis. 3 of these 6 patients were reoperated with a good effect, in 2 patients urine passage recovered 2-3 months after removal of transcutaneous puncture nephrostoma. In one child this technique was used for the first time at the third operation on vesicoureteral anastomosis. This prevented subsequent removal of the nephrostomic drain because of vesicoureteral reflux in the presence of severe irreversible retention changes of both ureters. The advantages of the above technique are reduced operative injury, no contraindications in primary operations and reoperations, feasibility in various forms of vesicoureteral anastomosis failure. This makes the technique of extravesical ureterocystoanastomosis rather promising. PMID- 12077824 TI - [Ultrasonic methods in diagnosing renal growths]. PMID- 12077825 TI - [Specific antigens in screening and monitoring patients with prostate cancer]. PMID- 12077826 TI - [Liver resection in locally-spread and metastatic kidney cancer]. AB - The postoperative outcome and survival were studied in patients operated for renal cancer with involvement of the liver. 9 patients have undergone radical nephrectomy and 12 patients--hepatic resections for direct hepatic involvement (2), synchronous (2) and metachronous (8) metastases of renal cell carcinoma. Right hemihepatectomy was performed in 2 and wedge resection in 10 cases. A complete resection was performed in 8 of 9 patients while one patient with direct hepatic invasion was found to have positive surgical margins. Postoperative lethality was absent but complications occurred in 6 patients: pancreatitis (1), pneumonia (3), hepatic abscess (1), hepatic and renal failure followed by GI bleeding (1). At follow-up, two patients died of progressive disease 4 and 68 months after the surgery and one was lost for follow-up. One patient with positive surgical margins is alive with pulmonary and liver metastases 16 months after surgery. Five patients are alive with no evidence of relapse 6, 10, 12, 19 and 56 months after the operation. Thus, the aggressive surgical approach is justified and should be considered in patients with renal cancer and hepatic involvement. PMID- 12077827 TI - [Correction of asthenovegetative disorders in the postoperative period in pulmonary tuberculosis patients]. AB - 58 patients with asthenia were observed during postoperative period after surgery treatment of tuberculosis. All of the patients were divided into two groups: 34 patients taking anti tuberculosis drugs and Stimol, and 24 patients taking only anti tuberculosis drugs. Following method where used to appreciate states of the patients: scales of Symptoms Check List--90, acid-base and gas state of blood, EKG, ECHOKG, KIG, Contactles Spirometry. RESULTS: Treatment with using of Stimol leads to quicker regress of asthenia to improvement in both cardiovascular and respiratory systems and normalization of gas state of blood. Effect of this medicine can be connected with positive influence on cardiac and respiratory functions of the body due to normalization vegetative prevision and improvement of metabolism in cells of drain and cardiomyocyties. PMID- 12077829 TI - [Minimally invasive valvuloplasty of the femoral vein in varicose disease]. AB - Length of incisions in varicose surgery is one of the main factor of surgery quality. Method and results of extravasal correction of femoral valves with own fascial tunics from mini-approach are presented. Endosurgical manipulations are performed within operative space created by hook-levator with endolight. It permitted to shorten incision of skin to 2-2.5 cm. Mini-invasive valvuloplasty was performed in 43 patients. The developed method meets esthetic criteria of modern phlebology. PMID- 12077828 TI - [Diagnosis and surgical treatment of bilateral pulmonary echinococcosis]. AB - Results of diagnosis and treatment of 205 patients with bilateral pulmonary echinococcosis were analyzed. 141 patients underwent two-sided surgeries: 77- staged operations with interval from 2 weeks to 3 months, 64--one-stage operations on both lungs. One-stage operations through thoracotomies were performed in 49 patients, through medial sternotomy--in 15. Mean hospital stay was 20 bed-days. Long-term results from 6 weeks to 15 years were studied in 60 patients. Stable curative effect was achieved in all the patients. PMID- 12077830 TI - [Treatment of patients with venous diseases of the lower extremities]. AB - Results of treatment of 355 patients with chronic venous insufficiency of the lower limbs were analyzed. In 169 patients with postthrombophlebitic disease, in 38 patients with postthrombophlebitic trophic ulcers and in 43 patients with varicose trophic ulcers the lymphotropic therapy was used (the study group). Control group consisted of 105 patients treated with traditional methods. Limphotropic therapy improved microcirculatory and metabolic processes in tissues that promotes faster cure of inflammation and edema and epithelization of trophic ulcers in patients with chronic venous insufficiency. PMID- 12077831 TI - [Comparative characteristics of efficacy of operations in ulcerous pyloroduodenal stenosis]. AB - 1107 patients with ulcerous pyloroduodenal stenosis (UPDS) were operated since 1968 to 2000. Duodenal ulcers caused stenosis in 1053 (95%) patients, gastric ulcers of types II and III--in 54 (5%). Various stomach resections were performed in 287 (92%) patients organ-saving operations--in 795 (71.8%) patients Longterm outcomes were studied in 797 (71.9%) patients, 491 (61.6%) of them were examined in hospital and 306 (38.4%) responded to questionnaires. Mean follow-up was 17 +/ 8.4 years. Early postoperative complications were in 138 (12.5%) patients, after resections--in 20.5%, after organ-saving operations--in 9.3%. General lethality after UPDS surgery was 0.6%, after organ-saving operations--0.2%, after resections--1.3%. Pathologic syndromes were seen in 31% patients. Recurrence of ulcer after organ-saving operations was revealed in 11% in remote terms. Demping syndrome was in 22.9% patients. Most often it was revealed after 2/3 stomach resection by Bilrot-II in Gofmeister--Finsterer (G--F) modification, less often- after SPV with duodenoplasty. The only operation preventing reflux-gastritis was economic stomach resection with Roux anastomosis and trunk vagotomy (TV + Er Roux). Good and excellent results by Visick scale were achieved in 85% patients with UPDS. Evaluation by D. Johnston scale demonstrated best results after SPV in duodenoplasty (238 points) and TV + ER Roux (272 points), the worst results- after B-1 (773 points) and B-II G--F (1124 points). Optimal results like healthy quality of life was seen after TV + ER Roux and SPV with duodenoplasty. PMID- 12077832 TI - [Surgical treatment of malignant tumors in the biliopancreatoduodenal zone]. AB - Results of surgical treatment of 381 patients with cancer of biliopancreatoduodenal zone associated with obstructive jaundice were analyzed. Mean level of bilirubinemia was 182 +/- 12 mcmol/l. Cholecystoanastomosis was created in the majority of cases (51.4%). Radical surgery was carried out in 31 patients. Postoperative complications were seen in 155 (41%) patients. Renal hepatic failure (28.1%) and purulent-septic complications (25.2%) were dominant. Lethality after radical operations was 12.9%, after palliative--15.7%. Inhibiting effect of autoplasma on phagocytosis and decrease of phagocytosis indexes 1.5-2 times are the risk factors of postoperative purulent-septic complications development. Discrete plasmapheresis reduces of postoperative purulent-septic complications rate. PMID- 12077833 TI - [Surgical tactics in penetrating wounds of the heart]. AB - Results of treatment of 43 patients with heart wounds are analyzed. Surgery must be performed at the same time as antishock and reanimation therapy. Success of operation depends on time of patient's transport to hospital, quality of performed surgery, postoperative treatment and severity of patient's state. Severity of patient's state is determined by location of the wound, its sizes and injuries of other organs. PMID- 12077834 TI - [Diagnosis and treatment of gunshot wounds to the chest]. AB - Experience of treatment of 86 patients with chest wounds are summarized. Diagnostic and curative algorithm of these patients are presented. Fenestration of the pericardium is proposed for diagnosis of heart wounds. This algorithm permitted to reduce early pleural complications rate from 55.1 to 22.1% and diagnostic mistakes--from 34.2 to 11.2%. PMID- 12077835 TI - [Long-term results of surgical treatment of patients with asymptomatic carotid artery stenosis]. AB - Results of treatment of 148 patients with asymptomatic carotid stenosis were analyzed in terms from 5 to 180 months. 75 (51%) patients were operated, 73 (49%) -were not operated. Non-operated asymptomatic patients with more than 90% stenosis and patients with prolonged (more than 18 mm) stenosis have the highest risk of cerebral events (4% annually and more). In long-term period (from the 70th month of follow-up) patients with the 70-90% stenosis are the group of high risk of cerebro-vascular symptoms. PMID- 12077836 TI - [Surgical tactics in gunshot wounds of the colon]. AB - Results of treatment of 76 patients with gunshot wounds of the colon received during local military conflicts and peacetime are analyzed. The patients were hospitalized within 30-120 min since the injury moment. Clinical examination, primary surgical treatment and laparoscopy were the best diagnostic methods. This order of diagnostic methods permitted to reduce time of examination from 60-80 to 20-40 min. Resection of injured part of the colon over the limits of contusion zone with "extraabdominal partial anastomosis" creation, Hartman's or Lachey's operation were the optimal surgery. In injury of contramesenteric part of the colon it makes sense to create loop colostoma by original method. The number of intraabdominal inflammatory complications decreased from 57 to 6.9%, lethal outcomes--from 17.6 to 6.9%. PMID- 12077837 TI - [Diagnosis and treatment of abdominal trauma in children]. AB - Treatment of 79 children with injuries of abdominal and retroabdominal organs was analyzed. Results of diagnosis and treatment were evaluated in 3 groups: group 1- 29 children treated conservatively; group 2--20 patients in whom total hemostasis was achieved during laparoscopy; group 3--30 children who have undergone open operations. Non-invasive and minimally invasive diagnostic methods permitted to avoid surgery in 46.4% patients. Efficacy of polymeric knitted netted material and Surgicell) for hemostasis in parenchymatous organs was demonstrated experimentally in rats. PMID- 12077838 TI - [Quality of life in patients after coloesophagoplasty, performed in childhood]. AB - Based on questionnaire, the authors analyze long-term results of coloesophagoplasty in 32 patients operated from 1990 to 2000. At the moment of operation age of the children ranged from 3 months to 14 years (5 years, on the average). 1 (3.1%) patient died at the age of 15 years, 9 years after esophageal plasty. Long-term results were evaluated in 31 respondents. Most respondents (81.3%) evaluate their state late after coloesophagoplasty as good and satisfactory. All the respondents take meal without assistance 74% patients don't keep a diet. Respondents with severe concomitant diseases (VACNERL--syndrome, bullous epidermolisis, CNS disorders) evaluate their state as unsatisfactory. In children after coloesophagoplasty falling behind growth (48%) and weight (62%) was revealed due to disease and low living standard. 89.5% respondents evaluate their quality of life as satisfactory. PMID- 12077839 TI - [Surgical treatment of hemorrhoids]. AB - Since 1994 in addition to standard operations for chronic and acute hemorrhoids in Krasnodar military hospital the device for suturing in removal of internal hemorrhoids has been applied. From 1994 to 2000 examination and treatment of 240 patients with hemorrhoids were carried out. In the study group (128 patients) hemorrhoidectomy was performed by the developed method, in control (112 patients) -by standard techniques. In early postoperative period significantly smaller quantity of complications were seen in the study group (8.4%) compared with control group (29.8%). Decrease of hospital stay and out-patient treatment was also seen. Relapses of the disease were not revealed in terms from one to six years after surgery. The device may be employed in simultaneous operations when concomitant anorectal diseases are present. The above results justify one-stage operations in combination of chronic hemorrhoids with anorectal diseases. The proposed device makes this surgery easier. PMID- 12077840 TI - [Problems and prospects anesthesiology]. AB - Anesthesiology is one of the most progressive medical science. Today's anesthesiologist is not only specialist in anesthetic management, he is also involved in pre- and postoperative treatment. The majority of urgent situations in hospital requires anesthesiologic assistance. But absence of prestige and low salary lead to low receipt of new specialists. The article also reviews the problems of training specialists, management, future prospects of modern anesthesiology. PMID- 12077841 TI - [Use of low frequency ultrasonic technology in preventing and treating purulent wounds of the lung and pleura]. PMID- 12077842 TI - [Senior surgeons at the front during World War II]. PMID- 12077843 TI - [Ultrasonic method of study and puncture biopsy in diagnosing thyroid nodules]. AB - Results of diagnosis of thyroid nodes in 404 patients are analyzed. Ultrasonic examination and US-assisted puncture biopsy were carried out before operations. Their results were compared with ones of morphologic study performed after surgery. Solitary nodes were diagnosed in 171 (42.3%) patients, multiple--in 233 (57.7%). The most specific ultrasonic symptoms of different forms of thyroid nodes were revealed. Thyroid cancer is imaged as hypoechogenic node with clear and unclear contour, heterogeneous structure, multiple zones of hyperechogenicity and calcification. Adenomas are presented as isoechogenic homogenic nodes with clear border and cystic cavities. Colloid nodes picture is the same. Rathen often different morphologic forms of nodes are similar, thus ultrasonic examination permits to detect morphologic structure of nodes hypothetically only. Sensitivity of puncture biopsy for goiter, adenomas and cancer was 87.1, 92.9 and 69.7% respectively. Cancer was not diagnosed before surgery in 30.3% cases when colloid goiter or adenomas were suspected. In these cases small cancer was located amidst colloid nodes and didn't differ by ultrasonic symptoms. Diagnosis of malignant tumor in nodular goiter was highly precise. Morphologic form of cancer was verified cytologically in majority of cases. PMID- 12077844 TI - [Steroids and other low molecular weight bioregulators]. PMID- 12077846 TI - [Novel triterpene glycosides from the sponge Ulosa sp]. AB - New triterpene glycosides, ulososides C, (20S,22S,23R,24S)-3 beta,22, 23 trihydroxy-3-O-(beta-D-glucopyranosyl)-32-nor-24-methyllanost- 8(9)-ene-30-oic acid, D, (20S,22S,23R,24S)-3 beta,22, 23-trihydroxy-3-O-(beta-D-N-acetyl glucosaminopyranosyl)-32-nor- 24-methyllanost-8(9)-ene-30-oic acid, and E, (20S,22S,23R,24S)-3 beta,22, 23-trihydroxy-3-O-(beta-D-glucuronopyranosyl-(1-->2) alpha-D- arabinopyranosyl-32-nor-24-methyllanost-8(9)-ene-30-oic acid, were isolated from an Ulosa sp. sponge. Their structures were determined by spectral methods and chemical transformations. Specific features of their structures are discussed. PMID- 12077845 TI - [Bromo- and iodo-containing alkaloids from marine microorganisms and sponges]. AB - The taxonomic distribution, structure, and biological activity of halogenated alkaloids isolated from marine microorganisms and sponges are reviewed. The structures of nearly 140 natural bromo- and iodo-containing alkaloids are shown. PMID- 12077848 TI - [NMR spectroscopy for studying structure and intramolecular dynamics of modified analogs of steroid hormones]. AB - Special features of the use of homo- and heteronuclear correlation methods of NMR in one and two dimensions for studying the spatial structure and intramolecular dynamics of modified analogues of steroid hormones (MASH) are considered. The application of these methods to the assignment of resonances in the high-field 1H NMR region and to the determination of the most stereospecifically important parameters, such as the vicinal constants of spin-spin coupling (3JH-H) and nuclear Overhauser effects (NOE), are discussed using the example of NMR studies of some estrogens and androgens at 300 MHz and on the basis of literature data. The most efficient combination of the methods and the necessary modification of each of them may be chosen considering the spectral and relaxation parameters of MASH in liquid medium, including the anisotropy of the overall diffusive motion. The characteristics of MASH are the wide use of correlations through long-range couplings (COSY-45 and DQF-COSY), the application of the 4,5JH-H constants for the determination of spatial structure, and the advantage of heteronuclear HSQC methods with and without 13C decoupling over the corresponding HMQC methods in both resolution and sensitivity. In the conformationally rigid MASH molecules, the anisotropy of the MASH diffusive motion in liquid adversely affects the determination of interproton distances by the calibrating processing method for the NOE difference and NOESY spectra: it results in both overestimated and underestimated distance values depending on the polar angle ratios of the reference and the determined distances. Under certain conditions, conformationally mobile MASH demonstrate the additional contribution of the scalar relaxation mechanism between the indirectly (scalarly) bound protons. This mechanism is responsible for the underestimated values of NOE and the corresponding errors in the distance determination. PMID- 12077847 TI - [Steroid compounds from the Far Eastern starfish Diplopteraster multipes]. AB - Sodium salt of (20R)-3 alpha,4 beta-dihydroxycholest-5-ene-21-yl sulfate and disodium salts of (20R)-4 beta-hydoxycholest-5-ene-3 alpha,21-diyl disulfate, (20R)-24-methylcholest-5,24(28)-diene-3 alpha,21-diyl disulfate, (20R)-24-methyl 5 alpha-cholest-24(28)-ene-3 alpha,21-diyl disulfate, (20R)-cholest-5-ene-3 alpha, 21-diyl disulfate, (20R)-5 alpha-cholestane-3 alpha,21-diyl disulfate, and (20R)-3 alpha-hydroxycholest-5-ene-2 beta,21-diyl disulfate were isolated from the far eastern starfish Diplopteraster multipes and characterized. These compounds differ structurally from sulfated polyhydroxysteroids in other starfish species. At the same time, they are typical secondary metabolites of Ophiuroidea and have some structural features characteristic of the ophiuroid-isolated steroids, namely the 3 alpha-hydroxy (or 3 alpha-sulfoxy) and 21-sulfoxy groups. These data support the opinion of some taxonomists that starfishes and ophiuroids are phylogeneteically related classes and are closer to each other than to other classes of the Echinodermata phylum. PMID- 12077849 TI - [Molecular model of binding of estradiol and 8-isoestradiol with estrogen alpha receptor]. AB - The complexes of the estrogen alpha-receptor with estradiol and 8-isoestradiol were comparatively analyzed. The computations of ligand-receptor complexes, carried out using the FLEXX program, allowed us to propose a model for the binding of the analogues of 8-isoestradiol. It was found that rings C and D of estradiol and 8-isoestradiol are similarly arranged in the ligand-binding pocket and coincide upon the superposition of the corresponding ligand-receptor complexes, whereas rings A and B do not coincide. The oxygen functions in position 17 of the estradiol analogues of both series coincide upon superposition, whereas the phenol 3-hydroxyl groups are 0.05 A apart. A comparison of the predicted biological properties of modified estradiol analogues of the natural and 8-isoseries with the available experimental data revealed their similarity. Synthetic 2-acetyl analogues of 8-isoestrogens were found to have no uterotropic activity, which is also consistent with the proposed model. PMID- 12077850 TI - [Synthesis and study of B-nor-8-isoanalogs of steroid estrogens]. AB - The study of the binding of estradiol B-nor-8-isonalogues to estrogen receptors from the rat uterus helped create the proposed model of the corresponding ligand receptor complexes. The use of this model ensured the choice of such micromodifications in this steroid group that sharply decreased their hormonal activity. By the example of 16,16-dimethyl-D-homo-B-nor-8-isoestrone, we demonstrated the possibility of the synthesis of the estrogen analogues devoid of uterotropic activity but retaining immunosuppressive activity. PMID- 12077851 TI - [Species and tissue distribution specificity of proteins binding 16alpha,17alpha cycloalkane derivatives of progesterone]. AB - The binding of [3H]progesterone and [3H] 16 alpha,17 alpha cycloalkanoprogesterones to proteins from rat, rabbit, and human uteri and other organs was studied. We found that 16 alpha,17 alpha-cycloalkanoprogesterone derivatives display affinities for the uterine progesterone receptors comparable with that of the natural hormone and no substantial species differences in the affinity. Rabbit uterus was found to have no proteins distinct from the progesterone receptor that specifically bind [3H] 16 alpha,17 alpha cycloalkanoprogesterones. At the same time, in the human uterus, we found another protein that binds some of these progesterone derivatives; it turned out to be similar to the protein from rat uterus. A similar protein with the same selectivity and affinity for steroids was also found in rat and human kidneys. Blood serum, liver, lung, and a number of other tissues were found to contain a protein of the third type that binds the same 16 alpha,17 alpha cycloalkanoprogesterones and exhibits submicromolar Kd values for these steroids and a very low affinity for progesterone. We speculated that the introduction of a bulky substituent adjacently to the 17 beta-side chain of progesterone could result in a change in the general biodynamics of the derivative including its transport, uptake, and accumulation in tissues, which may determine the selectivity of its effect. PMID- 12077852 TI - [Synthesis of O-substituted 6-oximes of 16alpha,17alpha-cyclohexanopregn-4-en 3,6,20-triones containing a tritium label at the 1,2-position]. AB - 6-O-(3-Methoxycarbonylpropyl)- and 6-O-(3-carboxypropyl)oximes of 16 alpha,17 alpha-cyclohexanopregn-4-ene-3,6,20-trione labeled by tritium in position 1,2 were synthesized. When using homogenous catalysts, the molar radioactivity of the resulting preparations was 1.5-1.7 PBq/mol. PMID- 12077853 TI - [Morphological changes in the rat uterine tissues exposed to pregna-D'-pentaran derivatives of progestins and anti-progestins]. AB - Changes in the uterus morphology of mature female rats were studied on the model of pseudopregnancy after treatment with the progestin 16 alpha,17 alpha cyclohexanoprogesterone (PR) and the antirpogestins 5 alpha(H)-16 alpha,17 alpha cyclohexano-4,5-dihydroprogesterone (APR1) and 5 beta(H)-16 alpha,17 alpha cyclohexano-4,5-dihydroprogesterone (APR2). The rats were preliminarily estrogenized with 17 beta-estradiol at a dose of 1 microgram/(animal day) for 4 days and then treated with PR at a dose of 0.2 mg/(animal day) for 14 days. The first group was then left without any treatment, whereas APR1 and APR2 were injected at the dose of 0.2 mg/(animal day) for 4 days to the animals of the second and the third groups, respectively. Light and electron microscopy of the uterus preparations demonstrated that the PR action provoked a complete pseudopregnancy picture characterized by the endometrium functionalization and the myometrium hypertrophy. Subsequent treatment with APR1 and APR2 caused the hypertrophy to cease, which had a more pronounced effect in the case of APR1. At the same time, some indications of the endometrium functionalization remained observable after treatment with APR1 and APR2. The specific binding sites for 3H labeled APR1 and APR2 were absent from the uterus cytosol for the rats gestagenized with PR. PMID- 12077854 TI - [Effect of mineralocorticoid analogs on Na(+),K(+)-dependent ATPase]. AB - The effect of 42 steroids of the 20-ketopregnane series with heterocycles fused in positions 16 alpha and 17 alpha on the activity of Na+,K(+)-activated ATPase from pig kidney was studied. It was shown that the studied compounds could be divided into two groups. The compounds from the first group stimulate the ATPase at low concentrations (1 x 10(-8)-1 x 10(-7) M) and inhibit it at high concentrations (1 x 10(-4) M). The second group of compounds stimulated the sodium pump at either concentration. This is explained by the cooperative action of the ATPase tetramer: after the reaction of its first binding site with the ligand, the tetramer changes the conformation and specificity of its other binding sites. Computer analysis of this series of compounds was carried out and a mathematical model of the dependence of their activities on the structure of their substituents was obtained with a high correlation coefficient and a satisfactory predictive power. This confirmed the structural similarity of the studied compounds with respect to their interaction with the ATPase binding sites. The method of descriptor analysis that was applied in this study is a new variant of approximation; it is based on the use of symbol variables as descriptors. PMID- 12077855 TI - [Novel synthesis of zymosterol]. AB - A modified scheme for the synthesis of zymosterol, one of the biosynthetically important yeast sterols, starting from from 3-benzoyloxyergosta-8(14),22-dien-15 one has been suggested. PMID- 12077856 TI - [Synthesis of C24-functionalized hydroxysterols]. AB - The syntheses of (24S)-24,25-epoxycholesterol, (24S)-hydroxycholesterol, and 24 ketocholesterol are described. The compounds belong to oxysterols, which can be considered to be the modulators of cholesterol metabolism. The asymmetric hydroxylation of desmosterol acetate according to Sharpless was used as the key reaction in the stereoselective introduction of functionality in position 24. PMID- 12077857 TI - Achieving physician buy-in through incentives. PMID- 12077858 TI - CMS clarifies when EMTALA applies. PMID- 12077859 TI - Computerized pathways should match written policy. PMID- 12077860 TI - Group closing GAP (Guidelines Applied in Practice) in heart attack care. PMID- 12077861 TI - Neonatal home program saves money, helps babies. PMID- 12077862 TI - DP, CM skills may stem bed-capacity problems. PMID- 12077863 TI - Health care seeks causes for discharge delays. PMID- 12077864 TI - Designing case management: the top 10 issues to consider. PMID- 12077865 TI - Managing international recruits. PMID- 12077867 TI - Facing the future. Interview by Steven Black. PMID- 12077866 TI - International recruitment. Quick fix or long-term investment. PMID- 12077868 TI - Organisational perspective. A pressure ulcer risk assessment and prevention audit: an RCN pilot. PMID- 12077869 TI - A catastrophe waiting to happen. Beating negligence in the independent sector. PMID- 12077870 TI - Developing your practice. PMID- 12077871 TI - How to add value in shared learning international alliances. PMID- 12077872 TI - [New markers improve the diagnostics of myocardial infarction]. PMID- 12077873 TI - [Why is the genome sequencing of meningococcus useful for us?]. PMID- 12077874 TI - [Nobel prize in medicine to transmitter researchers]. PMID- 12077875 TI - [Kondrad Reijo Waara Prize to Kari Eskola]. PMID- 12077876 TI - [Duodecim Information Award to Jari Sinkkonen]. PMID- 12077877 TI - [Diverticulosis of colon]. PMID- 12077878 TI - [Loop electrosurgical excision and follow-up of cervical dysplasia]. PMID- 12077879 TI - [Current treatment of psoriasis]. PMID- 12077880 TI - [A difficult impetigo or an autoimmune bullous dermatosis in a child?]. PMID- 12077881 TI - [Hemoptysis in a previously healthy woman in late pregnancy]. PMID- 12077882 TI - [Brief period of unconsciousness in a 3-year-old child]. PMID- 12077883 TI - [New antibacterial agents]. PMID- 12077884 TI - [What kind of new drugs will we get in the future and why?]. PMID- 12077885 TI - [Does collaboration inside the European Union improve Finnish drug safety?]. PMID- 12077886 TI - [Pharmaceutical industry and clinical drug research]. PMID- 12077887 TI - [Experiences of the drug approval processes in the European Union]. PMID- 12077888 TI - [Antihypertensive agents--principles of combination drug therapy]. PMID- 12077889 TI - [Statins--new mechanisms of action and new indications]. PMID- 12077890 TI - [New oncological drugs]. PMID- 12077891 TI - [Drug therapy of anxiety]. PMID- 12077892 TI - [Drug therapy of Alzheimer's disease]. PMID- 12077893 TI - [Diagnostics of myocardial infarction]. PMID- 12077894 TI - The biology of neurotropic viruses. PMID- 12077895 TI - Efferent system degeneration in vestibular ganglionitis. PMID- 12077897 TI - Antiviral therapy of vestibular ganglionitis. PMID- 12077896 TI - Neuroanatomy of the nerves in the temporal bone. PMID- 12077898 TI - Meatal ganglionitis: a pathologic correlate in idiopathic facial paralysis. PMID- 12077899 TI - Vestibular neuronitis: a viral neuropathy. PMID- 12077900 TI - Meniere's disease: a form of vestibular ganglionitis. PMID- 12077901 TI - The pathology of benign paroxysmal positional vertigo. PMID- 12077902 TI - A classification of recurrent vestibulopathy. PMID- 12077903 TI - Viral neuropathies in the temporal bone. Introduction. PMID- 12077904 TI - [Positron emission tomography and the Hungarian PET program]. AB - After a short introduction detailing the most basic features of the positron emission tomography (PET) the history of the preparation for the start of the Hungarian PET program is reviewed along with the results of the eight years activity of the Debrecen PET Centre. PMID- 12077905 TI - [Synthesis of radiopharmaceuticals for PET investigations]. AB - The PET radiopharmaceuticals are prepared on the spot in most cases due to the short lifetime of the isotopes used. The first step of this process is the isotope production by small cyclotrons. The synthons made from the isotopes react with the precursor of the given radiopharmaceutical. The target compound selected from the reaction mixture is ready for injection after purification, formulation and sterile filtration. In addition to [18F]-FDG, [11C]-methionine and [15O] butanol routinely used for diagnostic purposes in the PET Centre of the University of Debrecen, a number of other radiopharmaceuticals are synthesized for use in research from time to time. PMID- 12077906 TI - [Standard PET examination protocols]. AB - The authors briefly review the most important components, concepts and procedures of the PET method. They present the basic examination protocols and the special features of each type of investigation. A short summary of issues relating to the daily routine, the image processing and visualization as well as the data archiving is also included. PMID- 12077907 TI - [Spatial standardization of [15O]-butanol PET images using T1-weighted MRI data]. AB - Population averaged brain activation studies require the spatial standardization of the individual perfusion PET images. This procedure is usually supported by T1 weighted MRI images. The authors developed a segmentation technique to improve the automatized transformation of the individual MRI images into Talairach space. It was for the first time in Hungary that population averages of standardized T1 weighted MRI and [15O]-butanol PET images of the brain were created after validation of the procedure using data from 23 healthy volunteers. The newly developed method offers a solution for the automatized processing of primary data from brain activation experiments prior to statistical analysis. PMID- 12077908 TI - [Comparative analysis of quantitative evaluation methods for FDG-PET scans]. AB - The comparative analysis of two kinetic methods most frequently applied for the quantization of the results of FDG-PET brain scans was performed applying PET investigations and computer simulations. The analysis of the measured data and computer simulation demonstrated that the applicability of the results of SUV methods to estimate glucose metabolic rates (GMR) as calculated by the Phelps method (the most advanced method ) depended on the selected cerebral region. The distorted estimation was explained by the fact that the simpler models neglect dephosphorylation of the FDG-6P, and they also disregard the contribution of the intravascular activity to the tissue radioactivity. The simulations revealed the lack of a general and close positive correlation between SUV and GMR data; under special circumstances they might even change in an inverse manner. PMID- 12077909 TI - [Cost-effective PET scans in oncology]. AB - The authors have reviewed the financial considerations of oncological FDG PET examinations by the guidelines of the Health Care Financing Administration (USA). By critical assessment of large number of clinical investigations, the cost effectiveness of FDG PET scans has been confirmed in the following cases: differential diagnosis of solitary pulmonary nodule, diagnosis, staging and restaging of non-small cell lung cancer, colorectal cancer, malignant lymphomas, melanoma malignum, esophageal neoplasms and cancers of the head and neck. The role of this method in breast cancer is currently under intensive investigation. Due to the correct staging, PET examinations in these indications enable the clinicians to choose the optimal treatment ensuring the maximum probability of recovery and being cost-effective as unnecessary medical interventions become avoidable. PMID- 12077910 TI - [PET scan and double-independent pathologic investigations effectively support the detection of occult primary tumors]. AB - Following the failure of conventional diagnostic procedures, whole-body FDG-PET investigations were carried out in 42 metastatic cancer patients to localize occult primary carcinomas. During the clinical follow-up, the presence of malignant tumor was ruled out in 3 cases, and 2 patients originally believed to have carcinoma were confirmed to be suffering from a malignant hematological disease. These false diagnoses were associated with the use of imaging methods only (2 cases) or cytology only (1 case), lack of double, independent pathological investigations (2 cases) or immunophenotyping (2 cases) and the occurrence of an unrecognized rare tumor in a hospital with a small patient turnover (1 case). The discovered 11 occult primaries (4 lung, 3 breast, 2 hypopharynx and 1 base of the tongue carcinomas and 1 non-Hodgkin lymphoma) led to a 28% diagnostic efficacy of PET (11/39 malignant pathological reports). This efficacy is doubled (60%) if PET reveals < or = 5 malignant lesions and the locations of the pathological foci are tumor-specific. We suggest PET investigations in the search for occult primaries following a controlled pathological diagnosis and the failure of conventional diagnostic procedures. PMID- 12077911 TI - [Diagnostic possibilities of positron emission tomography in oncologic pulmonology]. AB - The authors present the possibilities of applying positron emission tomography (PET) in oncopulmonology. In addition to reviewing the literature, they share their own experience obtained during the diagnostic work-up and follow-up of twenty-three patients. The basic indications and the relevant properties of the most frequently used radiopharmaceuticals are discussed. Finally, the authors raise the question of cost-effectiveness of PET-investigations vs. conventional techniques, and suggest possible algorithms to include this non-invasive imaging method in the diagnostic work-up of patients with malignant or benign pulmonary diseases. PMID- 12077912 TI - [The role of PET scan in the investigation of the lymphatic spreading of Hodgkin's disease]. AB - The authors investigated the role of PET, as a non-invasive diagnostic method, in the analysis of lymphatic spreading of Hodgkin's disease (HD). Whole-body FDG scans were carried out in 71 patients along with [11C]-methionine examinations, if necessitated by inconclusive FDG results. Based on these findings involvement frequencies were calculated for each lymphatic region. The three most frequently involved lymphatic regions were the mediastinum (83.1%), the left cervical and left supraclavicular regions (78.9%) and the right cervical and right supraclavicular regions (76.1%). These data support the hypothesis that HD originates from the cervical or supraclavicular regions and reaches the distant sites by basically retrograde spreading in a non-random manner. The appropriate values of site involvement-rate were compared with those obtained by other authors based on pathologic staging and a good correlation was found. The high level of correspondence between these involvement-frequencies supported the general validity (i.e. valid for both treated and untreated cases) of the principles governing lymphatic spreading of HD. PMID- 12077913 TI - [The role of PET scan in the diagnosis of malignant melanoma]. AB - Comprehensive, accurate staging has a critical role in planning rational treatment strategies for patients with malignant melanoma (MM). In the present study the authors investigate the value of FDG PET in staging and restaging based on the investigation of 37 high-risk MM patients and compare the results with the one obtained by conventional imaging techniques (X-ray, US, CT, MR and bone scan). Thirty-nine whole body PET scans were carried out. The authors concluded that FDG PET had the highest sensitivity among the imaging methods in detecting distant metastases of MM. PMID- 12077915 TI - [Detection of recurrent head and neck cancer with positron emission tomography]. AB - Differentiation between recurrence and post-therapeutic lesions in patients with previously treated head and neck cancer can be a real diagnostic problem. The authors discuss the role of positron emission tomography in restaging of this disease by a retrospective analysis based on 20 scans of 17 patients. PET findings were correct in identifying tumors in 85% of patients (11/13) in the group of positive PET results. All negative PET findings (4/4) were also correct during the follow-up. PET results were validated in 88% (15/17) of the patients. In one case silent distant metastases were also detected. PET is a reliable method in restaging previously treated head and neck cancer patients as it can differentiate post-therapeutic changes from tumor relapse. PMID- 12077914 TI - [PET scan in patients with epipharyngeal tumors]. AB - Whole-body FDG PET examinations in 10 cases of epipharyngeal tumour (8 males, 2 females, mean age: 48 years) have been performed since January 1999. The PET examinations were aimed at the accurate staging, follow-up of the patients after the treatment, identification of recurrencies and localization of the unknown primary tumor. Functional imaging resulted in "upstaging" in 3 patients as compared to staging by the conventional diagnostic tools. Four additional patients with hyperplastic epipharyngeal tissue were investigated for occult primary cancer after negative results of multiple excisions, resulting in one case of primary epipharyngeal cancer. Correct staging, early detection of recurrencies, localization of occult primary tumor and the better post therapeutic assessment of epipharyngeal masses all facilitate a more reasonable therapeutic approach, which may improve survival results. PMID- 12077916 TI - [FDG PET scan of metastases in recurrent medullary carcinoma of the thyroid gland]. AB - Searching for metastases of medullary thyroid cancer (MTC), FDG PET was applied. PET results were compared with those of conventional diagnostic imaging procedures. After primary treatment, 52 MTC patients with elevated serum tumor marker levels and/or general symptoms (diarrhoea/flush) underwent radiological (CT/MRI), 131-iodine labeled metaiodo-benzylguanidine (MIBG) whole-body scintigraphy and FDG PET investigations. At least one pathological lesion was detected in 49 patients by PET, 35 by CT, 32 by MRI and 3 by MIBG. FDG PET was far superior to the other methods in identifying metastases in the supradiaphragmatic lymphatic regions. In the follow-up of MTC patients, FDG PET was more sensitive in localizing tumorous LN involvement, especially in the cervical, supraclavicular and mediastinal lymphatic regions, compared to other imaging modalities. PMID- 12077917 TI - [The role of FDG-PET scan in the diagnosis of pancreatic carcinoma]. AB - In 22 patients with focal pancreatic lesions, the values of different diagnostic modalities were analysed for the evaluation of malignancy. The CA 19-9 level was elevated in 4/5 malignant cases (sensitivity: 80%) and in 4/15 of benign cases (specificity: 73%). Hypodensity/hypoechogeneity was considered to be the characteristic sign of malignancy during CT/ultrasound (sensitivity: 100%, specificity: 50%/47%). ERCP was found to be specific but not sensitive enough and the rate of unsuccessful investigations was relatively high (4/22). The diagnostic value of FDG-PET was found to be superior to other diagnostic modalities (sensitivity 100%, specificity 88%). In cases with focal pancreatic hypoechogenic/hypodense lesions detected by CT or US in suspicion of malignancy, the FDG-PET should be the next step in the diagnostic strategy. PMID- 12077918 TI - [Positron emission tomography in the investigation of malignant testicular tumors]. AB - Thirty-three [18F]-FDG and ten [11C]-methionine (altogether 43) PET studies were performed in 37 (24 non-seminoma and 13 seminoma) patients. All results were assessed on the basis of histology (or cytology) or clinical follow-up. PET scan identified metastatic disease in 13 cases while 30 investigations resulted in a negative medical report. There were 3 false-positive cases and no false-negative results were obtained. The false-positive results were likely to occur due to FDG accumulation in benign lesions. There were no false-positive findings with the use of [11C]-methionine. Sensitivity, specificity and accuracy were 100%, 91% and 93%, respectively, using both tracers. PMID- 12077919 TI - [The PET scan in neuro-oncology--indications, differential diagnosis and clinical application]. AB - Results of 157 PET (positron emission tomography) studies in 102 patients with known or presumed intracranial (ic.) tumors are reviewed. In 106 studies [18F]-2 fluoro-2-deoxy-D-glucose (FDG) and in 51 cases [11C]-methionine (MET) was used as the PET tracer. CT and MRI imaging accompanied the PET studies in each case. Results of pre- and/or postoperative studies are compared. PET was concluded to have a fundamental role in the precise differential diagnosis in case of ic. space-occupying pathologies since it is capable of providing information that is not obtainable or may be equivocal based solely on CT and/or MRI investigations. However, it is also mandatory to be aware of the limits of PET. PMID- 12077920 TI - [Clinical positron emission tomography: brain imaging]. AB - PET produces images of physiological functions at molecular level which can be used to measure different vital processes, including blood flow and perfusion, glucose metabolism, oxygen utilization and receptor-ligand binding rates. Unlike anatomical imaging modalities, such as CT and MR, PET allows for the assessment of biochemical and physiological changes related to any function. Recently, the clinical indications for PET have increased dramatically. PET can reduce the overall cost of medical care, because it increases the certainty of diagnosis and clinical stage of disease, and therefore eliminates the expense of unnecessary testing or treatment. PMID- 12077921 TI - [The role of PET scan in the investigation of epileptic functional disorders]. AB - Systematic research is needed to evaluate the exact role of pathological factors in the determination of the extension of the hypometabolic area in partial epileptic patients. Together with structural damage, previous seizures, deafferentation and inhibitory mechanisms may contribute to the functional disorders. Benzodiazepine receptor studies showed that the density and binding ability of these receptors decreased in the area of epileptic functional disorder. Circumscribed hypermetabolism may appear during epileptic seizures or even more electrical discharges. The authors' PET studies aimed at presurgical evaluation showed that bilateral temporal hypometabolism occurred more frequently with right-sided seizure start. FDG-PET supported the localization of the pacemaker area both in temporal lobe epilepsies and in extratemporal epilepsies. This method proved instrumental in delineating the extension of the background pathology, too. The authors also demonstrated the strength of PET brain activation in mapping the hemispheric distribution of speech functions required in the planning of surgical interventions. The role of hippocampal sclerosis in temporal lobe hypometabolism was investigated and a short account is given of the observations relating to the relationship of hypermetabolism due to subclinical epileptic discharges and cognitive deficit symptoms. PMID- 12077923 TI - [Cerebral glucose metabolism in Down syndrome using positron emission tomography]. AB - Resting cerebral glucose metabolism was assessed by PET using FDG in eleven Down syndrome (DS) patients. Standardized uptake values (SUV) were determined on a pixel by pixel basis from the measured tissue FDG-activity data. The results showed that the mean value of the global FDG-uptake in the DS patients was not significantly different from the one obtained in the control group consisting of children and young adults. The standard deviation of SUV, however, was much higher in the DS group in almost all regions. The statistical parametric mapping (SPM) method was applied to compare the relative cerebral FDG accumulation pattern of the populations. Six regions (clusters) were found in which the glucose-uptake was higher in the DS patients than in the control group. The anatomical localization of these clusters was based on MRI investigations and brain-atlas technique. PMID- 12077922 TI - [PET and SPECT scans in autistic children]. AB - The authors have analyzed and compared the results of the 99mTc-ECD-SPECT and FDG PET examinations, performed in alert state, of 12 children suffering from infantile (9 subjects) or atypical (3 subjects) autism. In addition to frontally increased FDG metabolism, a decreased blood flow with left-sided dominance was found bifrontally and bitemporally in the infantile form (perfusion-metabolism mismatch). The regional differences in cortical FDG uptake were not significant in atypical autism, although both the blood flow and the metabolism of the thalami were decreased. Based on the results, the authors suggest that, beyond the usually inconclusive structural (CT or MR) examinations, cerebral blood flow SPECT and metabolic FDG-PET investigations may be useful in classifying the disease. PMID- 12077924 TI - [FDG-PET-scan in subarachnoid hemorrhage]. AB - Ruptured aneurysms cause subarachnoidal hemorrhage (SAH) that may or may not result in neurological sequelae. Acute SAH has been shown to reduce cerebral blood flow and metabolism. An important question is whether or not the SAH induced vasospasm without clinical symptoms leads to cerebral ischemia? Using postoperative [18F]-fluorodeoxyglucose positron emission tomography in patients with vasospasm diagnosed by transcranial Doppler without clinical changes, hypermetabolism was detected as an indicator of diffuse compensated ischemia in 6 cases. In 2 patients with neurological deficit reduced glucose metabolism was observed in the frontal and temporal-basal cortex. PMID- 12077925 TI - [Use of positron emission tomography in psychiatry]. AB - For the past years, substantial amount of experimental data has been published on the use of positron emission tomography in different psychiatric disorders. The different PET methods, which evaluate the whole-brain or regional metabolism, tissue perfusion or receptor density may help to identify the disorder specific changes in brain function and also better understand the underlying pathophysiology. In the clinical practice, PET plays an important role in the diagnosis and differential diagnosis of dementias and, presumably, in the near future the PET technique will be also extensively used in the clinical examination of other psychiatric disorders. PMID- 12077926 TI - [The role of PET scan among cardiologic imaging methods]. AB - The PET technique has been available for myocardial viability studies in Hungary since 1994. The early comparative results showed that FDG-PET identified more potentially reversible myocardial damages than [201Tl]-scintigraphy, defining the viability either by preserved metabolic activity of 50% or by metabolism perfusion mismatch. According to the follow up of revascularized patients in our cases, FDG-PET had a positive predictive value for functional recovery at 80%. In our opinion, it is the detection of glucose metabolism that predicts with the greatest accuracy segments of hibernating myocardial tissue that will recover after revascularization. Unlike dobutamine echocardiography, PET can detect viable ventricular segments in the myocardium of lost contractile material often seen in the histological studies of chronically ischemic myocardium. The FDG-PET method can be regarded as the gold standard of myocardial viability. PMID- 12077928 TI - [The [18F]-FNECA serves as a suitable radioligand for PET investigation of purinergic receptor expression]. AB - The well known and widely used P1 adenosine agonist, 5'-N-ethyl carboxamidoadenosine (NECA), was labelled with 18F isotope for the in vivo PET investigation of A1, A2 and A3 adenosine receptor expression. The precursor 2 [18F]fluoroethylamine was reacted with 2',3'-O-isopropylideneadenosine-5'-uronic acid. Specific activity of the [18F]-FNECA was (2.3 +/- 1.1) TBq/mmol (60 Ci/mmol). Dynamic PET measurements were carried out in rabbits to study the in vivo kinetics of the receptor saturation with the labelled ligand. The time dependent accumulation was followed up in the heart, lungs, liver, brain and testis. The radiotracer uptake was rapid and reached its maximum in less than two minutes in the heart and testes after v. injection of the radiopharmaceutical, while it took about 6 minutes in the brain, lungs and liver. High [18F]-FNECA accumulation was detected in the intestines, too. The specific binding of the [18F]-FNECA was tested in competition experiments in brain and heart sections using autoradiographic technique. The outlined synthesis provided sufficient amounts of [18F]-FNECA to map adenosine receptor expression under physiological conditions. PMID- 12077927 TI - [In vivo investigation of the A2A adenosine receptor distribution using the [11C] CSC radioligand]. AB - A 11C labeled selective adenosine A2A antagonist, (E)-8-(3-chlorostyryl)-1,3 dimethyl-7-[11C]-methylxanthine [(11C)-CSC] was prepared reacting (E)-8-(3 chlorostyryl)-1,3,-dimethylxanthine and [11C]-methyl iodide. A primary evaluation of [11C]-CSC as a potential tracer for mapping adenosine A2A receptors by positron emission tomography (PET) was also presented. Autoradiographic studies were carried out on Swiss mice. A high level accumulation of radioactivity was observed in the striatum and medulla oblongata in accordance with previous findings on the specific spatial distribution of A2A adenosine receptors. Dynamic PET studies on rabbits showed a fast brain uptake of CSC, reaching a maximum in less than 2 minutes. Competition experiments with the unlabeled ligand proved [11C]-CSC to bind specifically to the appropriate receptor. PMID- 12077929 TI - [Auditory event related potential and PET scan: a possibility for the comprehensive evaluation of cognition]. AB - The authors studied the parallel use of event related potential (ERP) and brain activation PET in nine healthy volunteers. The presence of P300 wave in the EEG recording was accompanied by a parallel increase in the blood perfusion of the anterior cingulate gyrus. Furthermore, the functional connectivity between cortical and subcortical structures was also present during the task. This method combines the fine temporal-resolution of ERP and spatial-resolution of PET, which allows for the complex examination of fast cognitive processes. This approach may be useful in the investigation of psychiatric disorders. PMID- 12077930 TI - [Movement-activated [15O]-butanol PET scan in patients with Parkinson disease]. AB - The regional cerebral perfusion has been studied by [15O]-butanol PET in bradykinetic patients with Parkinson's disease. In comparison with the rest condition significantly higher perfusion was detected in the primary sensorimotor cortex, parietal cortex, premotor cortex and supplementary motor area. A decrease of activity was observed in the lateral motor cortex, medial frontal cortex, dorsolateral prefrontal cortex and bilaterally in the head of the caudate nuclei. The method is capable of demonstrating the reorganisation of extrapyramidal movement regulation and the impact of medical and surgical treatment modalities. Based on the results conclusions can be drawn for the modes of action of the different treatment modalities. PMID- 12077931 TI - [Effect of flumazenil on the regional cerebral blood flow measured by [15O] butanol PET scan and transcranial Doppler ultrasonography]. AB - The authors investigated the role of the cBZD/GABA-A receptor-complex in the regulation of rCBF by simultaneously applying [15O]-butanol-PET and TCD in therapy-resistant TLE patients. It is known that the density of the cBZD/GABA-A receptors is reduced in the epileptogenic area of the temporal lobe, but there are no data available about the impact of this phenomenon on rCBF. FMZ is a well known BZD-receptor antagonist which can be used to inhibit the effect mediated by cBZD/GABA-A-receptors, offering a way to study the contribution of these receptors in the determination of rCBF. The authors examined the possible correlation between the rCBF values obtained by PET and the blood flow speed readings measured by TCD in MCAs. The results show that FMZ does not cause any change in rCBF on the side of the epileptogenic focus but it significantly raises rCBF (p < 0.001) in the intact brain tissue on the other side. The TCD measurements showed no significant differences between the blood flow in the MCAs supplying the affected and the contralateral lobe. PMID- 12077932 TI - [Effect of pathologic and induced peripheral vestibular balance disturbance on the central nervous system]. AB - The authors investigated the central projection of excitement in acute stage vestibular neuronitis using positron emission tomography. The changes in the pattern of regional cerebral blood flow caused by the disease were compared with the effect of cold caloric stimulation known to provoke similar signs. It was concluded that the involved brain regions overlapped each other only partially. The mismatch could be explained by the compensatory processes developing during the disease. These processes do not normally develop during the caloric vestibular stimulation because of its short duration. PMID- 12077933 TI - [Right prefrontal cerebral hemispheric activation by symmetrical carotid sinus baroreceptor stimulation]. AB - This study was performed to test the hypothesis of greater right hemispheric involvement in the processing of signals related to baroreceptor stimuli. Carotis sinus baroreceptors were stimulated by rhythmically decreasing air pressure in a neck chamber, and as a control the thorax was stimulated in a similar manner. Changes in regional cerebral blood flow (rCBF) were measured by positron emission tomography (PET). Baroreceptor stimulation resulted in rCBF increase in the right anterior-inferior prefrontal cortex (Brodmann areas [BA] 10/44/47) and bilaterally in BA 6/8. The authors conclude that, at least in some stages of baroreceptor information processing, the right hemisphere plays a greater role than the left one. PMID- 12077935 TI - [Sparfloxacin (Sparflo) in the treatment of urological infections]. AB - Sparfloxacin efficacy evaluation for the treatment of urogenital tract infections is presented. The trial was performed on 43 patients with chronic complicated infections of urogenital tract (pyelonephritis, prostatitis). Sparfloxacin (once daily for 7-14 days) was highly effective in the infection management according to clinical and bacteriological results (83.7 per cent and 78.1 per cent subsequently). PMID- 12077934 TI - [Comparison of linezolid and vancomycin in nosocomial pneumonia: results of the multicenter double-blind study]. AB - The results of multicenter, randomized, double-blind comparative study of linezolid and vancomycin efficacy, safety and tolerability in the treatment of nosocomial pneumonia are presented. The trial was performed on 69 patients. Clinical efficacy of linezolid was 83 per cent, of vancomycin--79 per cent. Bacteriological effect (pathogen eradication) was 83 per cent for linezolid group and 86 per cent for vancomycin group. During the study good clinical tolerability of linezolid was demonstrated along with lower side effects incidence and shortened recovery period when compared to vancomycin. PMID- 12077936 TI - [Antibiotic prophylaxis of postoperative complications in patients with burns]. PMID- 12077937 TI - [Sparfloxacin (a long-acting difluoroquinolone)--an antibacterial preparation with the broad spectrum activity]. PMID- 12077938 TI - [Micromycetes metabolites--inhibitors of growth and sterol biosynthesis in yeasts]. AB - Antifungal activity of micelial fungus metabolites (of genera Aspergillus, Penicillium, Fusarium, Stachybotris, Cladosporium, Alternaria, Gliocladium, Paecilomyces, Trichoderma etc.) was determined. It was shown that antifungal activity of some micromycetes is due to the formation of substances inhibiting sterols biosynthesis in eucaryote cells. Inhibitors of enzymes of sterols biosynthesis were isolated and their activity was investigated. It was shown, that isolated fungus inhibitors of sterols biosynthesis inhibited the growth of test-organism Rhodotorula rubra and decreased ergosterin level in yeast cells. The qualitative content of yeast cell sterols was not changed in the presence of fungus inhibitors. PMID- 12077939 TI - [Perspectives of use of moxifloxacin for treatment of urogenital infections]. PMID- 12077941 TI - Growth factors. PMID- 12077940 TI - [Immunotropic properties of exoglycan obtained from the marine micro-organism Vibrio alginolyticus]. AB - Immune system modulating activity of exoglycane isolated from culture media of Vibrio alginolyticus (strain 945-80) was investigated. The substance demonstrated stimulating activity on the humoral and cell immune system, potentiated phagocyte activity of macrophage and neutrophils, increased survival index of the animals infected by Gram-positive and Gram-negative bacteria. PMID- 12077942 TI - Muscle bound. PMID- 12077943 TI - Shock to the system. PMID- 12077945 TI - On the night shift in A&E. PMID- 12077944 TI - The danger zone. PMID- 12077946 TI - Bright spark. Interview by Julie Foxton. PMID- 12077947 TI - Developing safe i.v. habits. PMID- 12077948 TI - 'Keep your eye on the ball! is aimed at ensuring football players and supporters are aware of testicular cancer'. PMID- 12077949 TI - Patient in formation. 6. Summer skin care. PMID- 12077950 TI - Time is no excuse for having dirty hands. PMID- 12077951 TI - Basic first aid is not too much to ask. PMID- 12077952 TI - All aspects of sharps injuries need to be addressed. PMID- 12077953 TI - Religious beliefs and practices in acute mental health patients. AB - BACKGROUND: Holistic practice involves caring for the physical, psychological, social and spiritual needs of patients. Spiritual assessment is an important part of the patient's overall assessment. Religion is a component of spirituality and is supported by a recognisable formal system of beliefs. Religious beliefs might help patients in their search for meaning in life, especially in difficult circumstances such as during an episode of illness. A small, retrospective anonymous audit of patients' notes (n = 23) was undertaken to assess the recording of patients' religious affiliations, beliefs and practices in an acute mental health unit for older people. CONCLUSION: All individuals had a religious affiliation documented in their notes, but there was a lack of recorded information about their beliefs and practices. The findings showed that religious beliefs were only discussed with patients who had psychotic symptoms that had a religious content. Therefore, it would seem that patients on this unit are not having their spiritual and religious needs met. Staff training on religious awareness might help to address these issues. PMID- 12077954 TI - Pre-registration education: learning communities. AB - Changes in nurse education in the UK and the introduction of a new pre registration nursing programme have led to developments in education methods. This article describes the creation of learning communities at Thames Valley University as a means of adapting to the new curriculum. PMID- 12077955 TI - Prevention of infection: central venous catheters. AB - This article demonstrates how with set protocols, training and meticulous attention to detail, sepsis can be almost eradicated in central venous catheters. PMID- 12077956 TI - Diagnosis and management of urinary tract infection in children. AB - Urinary tract infection (UTI) causes significant illness, particularly in the first two years of life, and has the potential to cause permanent renal damage. Diagnosis of UTI in children is dependent on the collection of an uncontaminated freshly voided urine sample, which is key to the future management and follow-up of these patients. Prevention of possible long-term renal damage remains a priority. PMID- 12077958 TI - A question about leadership. PMID- 12077957 TI - Births: preliminary data for 2001. AB - OBJECTIVES: This report presents preliminary data for 2001 on births in the United States. U.S. data on births are shown by age, race, and Hispanic origin of mother. Data on marital status, prenatal care, cesarean delivery, and low birthweight are also presented. METHODS: Data in this report are based on more than 96 percent of births for 2001. The records are weighted to independent control counts of all births received in State vital statistics offices in 2001. Comparisons are made with 2000 final data. RESULTS: The number of births, the crude birth rate, and the fertility rate all declined slightly between 2000 and 2001. The number of births was down by less than 1 percent, the crude birth rate declined 1 percent to 14.5 per 1,000 population, and the fertility rate was down slightly to 67.2 births per 1,000 women aged 15-44 years. Teenagers were less likely to give birth in 2001; the teen birth rate continued to fall, dropping 5 percent between 2000 and 2001 to 45.9 births per 1,000 females aged 15-19 years, another record low. The teen birth rate has fallen 26 percent since 1991. The birth rate for teenagers 15-17 years fell 8 percent, and the rate for teenagers 18-19 years was down 4 percent for 2000-2001. Since 1991 rates have fallen 35 percent for teenagers 15-17 years, and 20 percent for teenagers 18-19 years. Birth rates for women aged 20-24 declined by 2 percent, whereas rates for women 25-44 years increased. Childbearing among women aged 40-54 years was stable. The birth rate for unmarried women decreased modestly to 44.9 births per 1,000 unmarried women 15-44 years in 2001, still remaining below the peak reached in 1994. The number of births to unmarried women was up very slightly, but births to unmarried teens were down. The proportion of women who began prenatal care in the first trimester of pregnancy improved slightly to 83.4 percent, but the rate of low birthweight held at 7.6 percent. The total cesarean delivery rate jumped 7 percent between 2000 and 2001 to 24.4 percent of all births, the highest level ever reported from this data source; the primary rate of cesarean deliveries rose 5 percent, and the rate of vaginal births after previous cesarean delivery tumbled 20 percent. PMID- 12077959 TI - The role of advanced practice nurses in community settings. PMID- 12077960 TI - From bedside to board room: interviews with nurse COOs and CEOs (Part 1). Interview by Alison P. Smith. PMID- 12077961 TI - Registered nurse pre-licensure education in California. PMID- 12077962 TI - Legal issues in performing patient safety work. PMID- 12077964 TI - Leading the leaders: the challenge of leading an empowered organization. PMID- 12077963 TI - A special treatment program for patients with sickle cell crisis. PMID- 12077965 TI - Implementing problem-based learning in ambulatory care. PMID- 12077966 TI - Health services research: a threatened foundation for the work of nurse executives. PMID- 12077967 TI - Low health literacy: what do your patients really understand? PMID- 12077968 TI - Satisfaction guaranteed? A sampling of strategies to keep experienced nurses on the job. PMID- 12077969 TI - The new threat to antibiotic resistance: why we need a health policy. PMID- 12077970 TI - Five nurses to be inducted into ANA's Hall of Fame. PMID- 12077971 TI - Georgia nurses develop network for disaster relief, emergency response. PMID- 12077972 TI - [Topical problems of trans-local pollution of areas]. AB - Translocal pollution is that resulted from the travel of effluents, discharges, and waste from one administrative area to another of the same country. Approaches to estimating interregional, intraregional, and intradistrict translocal pollution on the territory of the Russian Federation have not been developed so far. By using the calculation values, actual measurements with the background values borne in mind, the author assess intraregional pollution in the Volgograd Region, substantiated its criteria, and proposed a classification of translocal pollution. The proposed nomenclature, criteria, and classification of translocal pollution may be used to assess the transfer and impact of pollutions at the interregional, intraregional, and intradistrict levels on the territory of the Russian Federation. PMID- 12077974 TI - [Atmospheric pollution as a health risk factor in children and adolescence]. AB - Atmospheric pollution is an essential health risk factor. The present-day considerable rise in the incidence and prevalence of diseases among children and adolescents suggest that there is an excess of ecological pressure above the body's functional capacities of their adaptation. Children of the first year of life are the most vulnerable group. They are at the highest risk of death and disease with high technogenic loads (2.15-6.33 units). PMID- 12077973 TI - [Determination of critical parameters of environmental air pollution by the incidence of medical services utilization]. AB - The atmospheric environment in the town of Solikamsk was estimated by the daily measurements of 9 compounds at 6 stations of the Russian Committee of Hydrometereology during a year. The health status of the children under 14 years was evaluated by the rate of referrals for emergency health care. The results of stimulation of the impacts of short-term variations in the magnitude of ambient air pollution on the rate of referrals suggest that there is a rather clear correlation of the rate of referrals with gradient of the air concentrations of pollutants rather than with the absolute values. The indicators of direct toxic effects on respiratory organs are H2S, Cl, SO2, HCl and those on the skin are formaldehyde, SO2, HCl. PMID- 12077975 TI - [Methodological aspects of hygienic evaluation of small-sized plants safety for biological purification of household waste waters]. AB - The paper deals with the studies of the efficiency of four small-sized plants for biological purification of household waste waters (WW), which were conducted by using a complex of sanitary criteria, such as organoleptic, sanitary-and-chemical and sanitary-and-microbiological parameters of water quality of water objects, such as purified WW reservoirs, and presents a comparative assessment of their efficiency. The used methodological scheme is shown to provide an objective evaluation of the sanitary efficiency of the equipment under test with an compulsory set of regulated and additional integral parameters that should be regarded in combination. It is recommended that preference should be given to WW purification technologies that are effective in removing the environmentally hazardous chemical substances used in household chemistry. PMID- 12077976 TI - [Methods for purifying emissions in the boilers of thermoelectric plants from benz(a)pyrene]. AB - The researchers of the Republican Cancer Research Center of the Ministry of Health of the Republic of Uzbekistan have been monitoring the levels of the carcinogenic agent benz(a)pyrene (B(a)P) in the emissions by thermoelectric plants. Measuring has indicated that the effluent levels of B(a)P are related to the type of fuel put in the boilers and to the parameters of their use. The burning of gas in the furnaces results in air emissions of B(a)P at concentrations of as high as 50 ng/m3 and that of coal does as high as 300 ng/m3. Addition of fuel oil to coal or gas burning elevates the levels of B(a)P up to 450 and 180 ng/m3, respectively. Greater oxygen feed in the furnaces of boilers and their use at the maximum capacities (loads) cause a decrease in B(a)P in the waste gases. Recommendations on the optimum regimens of exploitation factors are given to lower environmental B(a)P levels, which must in turn reduce the incidence of cancer diseases. PMID- 12077977 TI - [Beta-carotene prevention of immune disorders in workers engaged in the fire copper refining]. AB - The paper deals with the studies of the human status and immunological responsiveness of workers engaged in copper-smelting industry. The examinees were found to more frequently develop lower environmentally induced immunological responsiveness followed by leuko- and lymphopenia, higher lymphocytic CD4+/CD8+ ratios, phagocytic dysfunction in peripheral blood. Preventive use of beta carotene prevented negative changes in immunological parameters. PMID- 12077978 TI - [Sanitary-and-epidemiological human well-being strategy under the conditions of socioecological development of the Russian Federation for the period until 2010]. PMID- 12077979 TI - [The specific features of microbiocenosis and fraction-dispersion composition of bacterial aerosol particles in industrial premises]. PMID- 12077980 TI - [Biological value of the average daily diets of elderly and old individuals in Uzbekistan]. AB - Nutrition was studied in the elderly (60-74 years), old individuals (75-90 years), and long livers (above 90 years) of Tashkent and the Kashkadarya Region by estimating the biological values of their diets and the biochemical parameters of vitamin C metabolism. Low biological values of actual diets, low vitamin C supply were found in the elderly and old individuals. Alimentary correction of the biochemical parameters of vitamin C metabolism was ascertained to be possible by enhancing the biological value of their diets by taking into account the national peculiarities of nutrition and relevant dietary technologies. PMID- 12077981 TI - [Hygienic evaluation of schoolchildren's nutrition in Riazan' (according to questionnaire data)]. PMID- 12077982 TI - [Correction of schoolchildren's diets with biologically active additives]. AB - The actual nutrition of schoolchildren in the Nizni-Novgorod district is imbalanced due to the deficiency of protein and vitamins and to the high contents of fats and carbohydrates. The authors provide evidence for a combined preparation to correct the children's diets, which incorporates animal protein, biologically active plant additives, and egg-shell calcium. The use of the preparation in combination with liquid bifidumbacterin contributes to increases in morphofunctional parameters in adolescents. PMID- 12077983 TI - [Vegetable levels of heavy metals in the Tomsk Region]. PMID- 12077985 TI - [Age-specific changes in anthropometric values of Krasnoiarsk children]. AB - The physical development of Krasnoyarsk children was studied. The anthropometric values of children aged 7-16 years were shown to become stable by the mid-1980s. The specific features of today's children versus those of the 1970s is an increase in the nonuniformity of physical development in puberty. PMID- 12077986 TI - [Factors influencing morbidity rates in preschool children in the city of Orsk]. AB - The comparative contribution of environmental factors to morbidity among the preschoolchildren of the town of Orsk was studied by applying currently available methods. Great morbidity is favoured by toxicosis during maternal second-half pregnancy, the poor health status of neonatal babies at birth and by the end of the first year of life, going to the nurseries where vegetable and calorie diets are not observed, the location of kindergartens in the areas with high traffic, high ambient air pollution with sulfuric anhydride and acid, and with substandard drinking water quality. Lower morbidity in children is in turn promoted by greater quantities of vegetables and fruits in the meals both at home and in the nurseries and kindergartens, over 2-month breast feeding, no adverse working conditions in their parents, as well as implementation of conditioning procedures in children. PMID- 12077984 TI - [Evaluation of immunological responsiveness in female students during an academic year]. AB - Examining 17 untrained female students and 110 trained ones indicated that moderate exercises as aerobic ones did not exert a great impact on most parameters of cellular and humoral immunity. Exclusions were higher absolute counts of B lymphocytes in the trained students than those in the untrained ones at the end of an academic year (in spring). Despite their activity, all the female students were found to have seasonal variations in some immunological parameters. Among other things, the content of immunoglobulins (primarily classes G and A) dropped in winter and particularly in spring as compared to autumn; on the contrary, the parameters of phagocytosis significantly increased in winter versus autumn, then fell in spring months as compared to winter; vive versa, the count of circulating immune complexes substantially reduced in winter as compared with autumn, then showing a trend for an increase in spring, at the end of an academic year. PMID- 12077987 TI - [Analysis of causes of allergic diseases in the children of the Republic of Tatarstan]. AB - The incidence of allergic diseases in children from poor ecological areas in the towns Nizhnekamsk, Kazan, and Almetyevsk, and in the rural settlement of Kukmor was studied. Allergic skin diseases in the children were shown to prevail in the structure of allergies despite the place of their residence. In the rural area (Kukmor), the incidence was 5-10 times lower than that in the urban areas. A correlation analysis has shown that there is a definite association of the incidence of allergic diseases and their prevalence with the characteristics of ambient air pollution. The findings suggest that measures reducing allergic diseases and environment pollution in these towns should be introduced. PMID- 12077988 TI - [Criteria for choosing priority parameters of chemical water contamination of socio-occupational monitoring]. PMID- 12077989 TI - [Methods of interlaboratory comparative trials in the epidemiological surveillance center of the Russian Federation]. PMID- 12077990 TI - [Assessment of the efficiency of informational activities in primary prevention of HIV infection]. PMID- 12077991 TI - [Virological study of different waters in the Moscow Region]. PMID- 12077992 TI - [Problems in monitoring the air levels of naphthalene]. PMID- 12077993 TI - [Methods for the determination of the levels of glutathione and the activity of glutathione peroxidase in red blood cells]. PMID- 12077994 TI - [Determination of free-radical methods for the evaluation of the effects of polychlorinated dioxins and furans on human health]. PMID- 12077995 TI - [Tests of milk for Mycobacterium tuberculosis by using the polymerase chain reaction]. PMID- 12077996 TI - [High-performance liquid chromatographic determination of aldehydes in biological media]. PMID- 12077997 TI - [Determination of renal asbestos elimination]. PMID- 12077998 TI - Nitrogen in aquatic ecosystems. AB - Aquatic ecosystems respond variably to nutrient enrichment and altered nutrient ratios, along a continuum from fresh water through estuarine, coastal, and marine systems. Although phosphorus is considered the limiting nutrient for phytoplankton production in freshwater systems, the effects of atmospheric nitrogen and its contribution to acidification of fresh waters can be detrimental. Within the estuarine to coastal continuum, multiple nutrient limitations occur among nitrogen, phosphorus, and silicon along the salinity gradient and by season, but nitrogen is generally considered the primary limiting nutrient for phytoplankton biomass accumulation. There are well-established, but nonlinear, positive relationships among nitrogen and phosphorus flux, phytoplankton primary production, and fisheries yield. There are thresholds, however, where the load of nutrients to estuarine, coastal and marine systems exceeds the capacity for assimilation of nutrient-enhanced production, and water quality degradation occurs. Impacts can include noxious and toxic algal blooms, increased turbidity with a subsequent loss of submerged aquatic vegetation, oxygen deficiency, disruption of ecosystem functioning, loss of habitat, loss of biodiversity, shifts in food webs, and loss of harvestable fisheries. PMID- 12077999 TI - The globalization of nitrogen deposition: consequences for terrestrial ecosystems. AB - The sources and distribution of anthropogenic nitrogen (N), including N fertilization and N fixed during fossil-fuel combustion, are rapidly becoming globally distributed. Responses of terrestrial ecosystems to anthropogenic N inputs are likely to vary geographically. In the temperate zone, long-term N inputs can lead to increases in plant growth and also can result in over enrichment with N, eventually leading to increased losses of N via solution leaching and trace-gas emissions, and in some cases, to changes in species composition and to ecosystem decline. However, not all ecosystems respond to N deposition similarly; their response depends on factors such as successional state, ecosystem type, N demand or retention capacity, land-use history, soils, topography, climate, and the rate, timing, and type of N deposition. We point to some of the conditions under which anthropogenic impacts can be significant, some of the factors that control variations in response, and some areas where uncertainty is large due to limited information. PMID- 12078000 TI - Reactive nitrogen and human health: acute and long-term implications. AB - Reactive-nitrogen (Nr) has a wide variety of beneficial and detrimental effects on human health. The most important of the beneficial effects are increasing global and regional food supplies and increased nutritional quality of available foods. However, lack of adequate dietary intake of amino acids and proteins is a serious cause of malnutrition when food supplies are inadequate because of poverty, drought, floods, wars, and displacements of people as refugees. There is sufficient, though limited, quantitative data indicating that increased circulation of Nr in the environment is responsible for significant human health effects via other exposure pathways. Nr can lead to harmful health effects from airborne occupational exposures and population-wide indoor and outdoor air pollution exposures to nitrogen dioxide and ozone. Nr can also affect health via water pollution problems, including methemoglobinemia from contaminated ground water, eutrophication causing fish kills and algal blooms that can be toxic to humans, and via global warming. The environmental pollutants stemming from reactive nitrogen are ubiquitous, making it difficult to identify the extent to which Nr exerts a specific health effect. As all populations are susceptible, continued interdisciplinary investigations are needed to determine the extent and nature of the beneficial and harmful effects on human health of nitrogen-related pollutants and their derivatives. PMID- 12078001 TI - Nitrogen and food production: proteins for human diets. AB - Nitrogen was the most commonly yield-limiting nutrient in all pre-industrial agricultures. Only the Haber-Bosch synthesis of ammonia broke this barrier. The rising dependence on nitrogenous fertilizers, which represents the largest human interference in the biospheric N cycle, has two different roles. In affluent nations it helps to produce excess of food in general, and of animal foods in particular, and it boosts agricultural exports. But for at least a third of humanity in the world's most populous countries the use of N fertilizers makes the difference between malnutrition and adequate diet. Our understanding of human N (protein) needs has undergone many revisions and although some uncertainties still remain it is clear that average protein intakes are excessive in rich countries and inadequate for hundreds of millions of people in Asia, Africa, and Latin America. More dietary protein will be needed to eliminate these disparities but the future global use of N fertilizers can be moderated not just by better agronomic practices but also by higher feeding efficiencies and by gradual changes of prevailing diets. As a result, it could be possible to supply adequate nutrition to the world's growing population without any massive increases of N inputs. PMID- 12078002 TI - Agroecosystems, nitrogen-use efficiency, and nitrogen management. AB - The global challenge of meeting increased food demand and protecting environmental quality will be won or lost in cropping systems that produce maize, rice, and wheat. Achieving synchrony between N supply and crop demand without excess or deficiency is the key to optimizing trade-offs amongst yield, profit, and environmental protection in both large-scale systems in developed countries and small-scale systems in developing countries. Setting the research agenda and developing effective policies to meet this challenge requires quantitative understanding of current levels of N-use efficiency and losses in these systems, the biophysical controls on these factors, and the economic returns from adoption of improved management practices. Although advances in basic biology, ecology, and biogeochemistry can provide answers, the magnitude of the scientific challenge should not be underestimated because it becomes increasingly difficult to control the fate of N in cropping systems that must sustain yield increases on the world's limited supply of productive farm land. PMID- 12078003 TI - Reducing global NOx emissions: developing advanced energy and transportation technologies. AB - Globally, energy demand is projected to continue to increase well into the future. As a result, global NOx emissions are projected to continue on an upward trend for the foreseeable future as developing countries increase their standards of living. While the US has experienced improvements in reducing NOx emissions from stationary and mobile sources to reduce ozone, further progress is needed to reduce the health and ecosystem impacts associated with NOx emissions. In other parts of the world, (in developing countries in particular) NOx emissions have been increasing steadily with the growth in demand for electricity and transportation. Advancements in energy and transportation technologies may help avoid this increase in emissions if appropriate policies are implemented. This paper evaluates commercially available power generation and transportation technologies that produce fewer NOx emissions than conventional technologies, and advanced technologies that are on the 10-year commercialization horizon. Various policy approaches will be evaluated which can be implemented on the regional, national and international levels to promote these advanced technologies and ultimately reduce NOx emissions. The concept of the technology leap is offered as a possibility for the developing world to avoid the projected increases in NOx emissions. PMID- 12078004 TI - Reactive nitrogen and public policies for environmental protection. AB - A core goal of both US and European pollution control policies has been to establish rules and regulations pertaining to the movement of reactive-nitrogen (Nr) through the environment. This is manifest in US federal legislation such as the Clean Air Act Amendments and the Clean Water Act Amendments and in various protocols of the United Nations Economic Commission for Europe (UNECE) and its Convention on Long-Range Transboundary Air Pollution (CLRTAP). In this paper, we begin by reviewing the two US laws and their effectiveness and make some comparisons with the approaches used mainly in Europe by the UNECE and CLRTAP. Next we use the Mississippi drainage/Gulf of Mexico hypoxia case study to highlight the importance of applying a "systems approach" to address the reactive nitrogen problem at the regional scale. After briefly posing a number of unanswered questions related to nitrogen control policies, we conclude by sketching a blueprint for future actions related to the development of improved policies to regulate reactive nitrogen. PMID- 12078005 TI - Nutrient management in food production: achieving agronomic and environmental targets. AB - The notion of management has undergone many changes during the past century. Nowadays, management is perceived as "specialized activity to achieve targets." Skill in management is the single most important factor determining the economic and environmental performance of agroecosystems. Nutrient management is "management of nutrients to achieve agronomic and environmental targets;" it requires proper understanding of nutrient cycling, site- and farm-specific guidelines and technology, and often direct coaching. These activities are diverse and complicated, especially in mixed farming systems that involve both crop and animal production. To be effective, economic and environmental targets must be coherent, flexible, and controllable. They also must be defined and implemented quantitatively at strategic, tactical, and operational levels. Data from farms in Poland and The Netherlands are used to show how economic incentives, provided through governmental policies and measures in both countries, can improve nutrient-use efficiency by a factor of 2 on many intensively managed mixed farming systems. PMID- 12078006 TI - Nitrogen fertilizers: meeting contemporary challenges. AB - At 81.7 million tonnes (Mt), commercial fertilizer nitrogen (N) accounts for approximately half of all N reaching global croplands today and supplies basic food needs for at least 40% of the population. The challenge is to continue to help meet that need while minimizing the risk of negative environmental impacts through improved N-use efficiency. Fertilizer-N efficiency on corn in the US has increased more than 30% over the last 20 years, but additional progress can be made for corn and other crops. Current N efficiency and productivity are generally lower in most of Asia than in North America, but they are improving. The fertilizer industry recognizes its crucial role in meeting basic human needs, now and in the future. It stands ready to meet the challenge of adopting new practices and technologies that will allow it to do so with greater efficiency and in a way that not only sustains life, but also sustains the quality of life. PMID- 12078007 TI - Decreasing reliance on mineral nitrogen--yet more food. AB - Higher crop production normally demands higher nutrient application rates and consequently increased mineral nitrogen use. With food demand for 2030 estimated around 2800 mill. tonnes (t) yr-1, the corresponding mineral N consumption figure is 96 mill. t (78 mill. t yr-1 in 1995/1997). Global-level mineral N losses to the environment from mineral fertilizer use are currently 36 mill. t yr-1, worth USD 11,700 mill. and with adverse environmental impacts. However, innovative fertilizer-use efficiency (FUE) technologies enable increased production with a less than a proportionate increase in mineral-N use. Moreover, nitrogen-nutrient supplies can be augmented through improvements in agricultural production systems and in the exploitation of alternative sources such as biological nitrogen fixation (BNF). By 2030, with adequate policy, technology transfer, research and investment support, the on-farm adoption of BNF and FUE technologies could generate savings of 10 mill. t yr-1 of mineral N, worth USD 3300 mill. PMID- 12078008 TI - Energy, industry and nitrogen: strategies for decreasing reactive nitrogen emissions. AB - Nitrogen oxides are released during atmospheric combustion of fossil fuels and biomass, and during the production of certain chemicals and products. They can react with natural or man-made volatile organic compounds to produce smog, or else can be further oxidized to produce particulate haze, or acid rain that can eutrophy land and water. The reactive nitrogen that begins in the energy sector thus cascades through the atmosphere, the hydrosphere and soils before being eventually partially denitrifed to the global warming and stratospheric ozone depleting gas nitrous oxide or molecular nitrogen. This paper will suggest how an economic analysis of the nitrogen cycle can identify the most cost-effective places to intervene. Nitrogen oxides released during fossil-fuel combustion in vehicles, power plants and heating boilers can either be controlled by add-on emission control technology, or can be eliminated by many of the same technical options that lead to carbon dioxide reduction. These integrated strategies also address sustainability, economic development and national security issues. Similarly in industrial production, it is more effective to focus on redesigning industrial processes rather than on nitrogen oxide pollution elimination from the current system. This paper will suggest which strategies might be utilized to address multiple benefits rather than focusing on single pollutants. PMID- 12078009 TI - NitroGenius: a nitrogen decision support system. A game to develop the optimal policy to solve the Dutch nitrogen pollution problem. AB - A nitrogen decision support system in the form of a game (NitroGenius) was developed for the Second International Nitrogen Conference. The aims were to: i) improve understanding among scientists and policy makers about the complexity of nitrogen pollution problems in an area of intensive agricultural, industrial, and transportation activity (The Netherlands); and ii) search for optimal policy solutions to prevent pollution effects at lowest economic and social costs. NitroGenius includes a model of nitrogen flows at relevant spatial and temporal scales including emissions of ammonia and nitrogen oxides and contamination of surface- and groundwaters. NitroGenius also includes an economic model describing relationships for important sectors and impacts of different nitrogen control measures on Gross Domestic Product (GDP), unemployment, energy use, and environmental costs. About 50 teams played NitroGenius during the Second International Nitrogen Conference. The results show that careful planning and selection of abatement options can solve Dutch nitrogen problems at reasonable cost. PMID- 12078010 TI - Reactive nitrogen: too much of a good thing? PMID- 12078012 TI - The European nitrogen case. AB - The N budget for Europe (excluding the former Soviet Union) indicates that the 3 principal driving forces of the acceleration of the European N cycle are fertilizer production (14 Mt (mill. tonnes) N yr-1), fossil fuel combustion and other industry (3.3 Mt N yr-1) and import of N in various products (7.6 Mt N yr 1). The various leaks of reactive N species from European food, energy and industrial production systems are estimated and their effects on human health and terrestrial and aquatic ecosystems are assessed. Future European environmental policy measures to close the N cycle and to reduce leaks of reactive N can best focus on the three major driving forces, taking into consideration the possible consequences in the N cascade. Critical loads may be useful tools in determining N-emission ceilings and developing integrated policies for regulating N flows such as fertilizer use and imports and N levels. PMID- 12078011 TI - Reactive nitrogen and the world: 200 years of change. AB - This paper examines the impact of food and energy production on the global N cycle by contrasting N flows in the late-19th century with those of the late-20th century. We have a good understanding of the amounts of reactive N created by humans, and the primary points of loss to the environment. However, we have a poor understanding of nitrogen's rate of accumulation in environmental reservoirs, which is problematic because of the cascading effects of accumulated N in the environment. The substantial regional variability in reactive nitrogen creation, its degree of distribution, and the likelihood of increased rates of reactive-N formation (especially in Asia) in the future creates a situation that calls for the development of a Total Reactive Nitrogen Approach that will optimize food and energy production and protect environmental systems. PMID- 12078013 TI - The Asian nitrogen cycle case study. AB - We analyzed nitrogen budgets at national and regional levels on a timeline from 1961-2030 using a model, IAP-N 1.0. The model was designed based upon the Inter governmental Panel on Climate Change (IPCC) methods using Asia-specific parameters and a Food and Agriculture Organization of the United Nations (FAO) database. In this paper we discuss new reactive-nitrogen and its various fates, and environmental nitrogen enrichment and its driving forces. The anthropogenic reactive nitrogen of Asia dramatically increased from approximately 14.4 Tg N yr 1 in 1961 to approximately 67.7 Tg N yr-1 in 2000 and is likely to be 105.3 Tg N yr-1 by 2030. Most of the anthropogenic reactive-nitrogen has accumulated in the environment. We found that an increasing demand for food and energy supplies and the lack of effective measures to improve the efficiency of fertilizer nitrogen use, as well as effective measures for the prevention of NOx emissions from fossil-fuel combustion, are the principal drivers behind the environmental nitrogen-enrichment problem. This problem may be finally solved by substituting synthetic nitrogen fertilizers with new high-efficiency nitrogen sources, but solutions are dependent on advances in biological technology. PMID- 12078015 TI - Nitrogen and nature. AB - Anthropogenic changes to the global N cycle are important in part because added N alters the composition, productivity, and other properties of many natural ecosystems substantially. Why does added N have such a large impact? Why is N in short supply in so many natural ecosystems? Processes that slow the cycling of N relative to other elements and processes that control ecosystem-level inputs and outputs of N could cause N supply to limit the dynamics of ecosystems. We discuss stoichiometric differences between terrestrial plants and other organisms, the abundance of protein-precipitating plant defenses, and the nature of the C-N bond in soil organic matter as factors that can slow N cycling. For inputs, the energetic costs of N fixation and their consequences, the supply of nutrients other than N, and preferential grazing on N-fixers all could constrain the abundance and/or activity of biological N-fixers. Together these processes drive and sustain N limitation in many natural terrestrial ecosystems. PMID- 12078014 TI - Nitrogen use in the United States from 1961-2000 and potential future trends. AB - Nitrogen inputs to the US from human activity doubled between 1961 and 1997, with most of the increase in the 1960s and 1970s. The largest increase was in use of inorganic N fertilizer, but emissions of NOx from fossil-fuel combustion also increased substantially. In 1961, N fixation in agricultural systems was the largest single source of reactive N in the US. By 1997, even though N fixation had increased, fertilizer use and NOx emissions had increased more rapidly and were both larger inputs. In both 1961 and 1997, two thirds of reactive N inputs were denitrified or stored in soils and biota, while one third was exported. The largest export was in riverine flux to coastal oceans, followed by export in food and feeds, and atmospheric advection to the oceans. The consumption of meat protein is a major driver behind N use in agriculture in the US Without change in diet or agricultural practices, fertilizer use will increase over next 30 years, and fluxes to coastal oceans may increase by another 30%. However, substantial reductions are possible. PMID- 12078016 TI - [Alzheimer dementia: positive diagnosis and prevention]. PMID- 12078018 TI - [Chronic neurotoxic damage in ecstasy (MDMA) users. Review of the current state of research]. AB - The popular dance drug ecstasy (3,4-methylenedioxymethamphetamine, or MDMA, and some analogues) causes selective and persistent neurotoxic damage of the central serotonergic system in laboratory animals. Serotonin plays a role in numerous functional systems in the CNS. Consequently, various abnormalities including psychiatric, vegetative, neuroendocrine, and cognitive disorders might be expected in humans following damage of the central serotonergic system. In recent years, the questions of possible functional disorders following ecstasy-induced neurotoxicity were addressed in several cross-sectional studies with drug users. In this review we summarize and evaluate the quality of design of these studies. Despite large methodological problems, evidence accumulates in favor of persisting brain damage in ecstasy users resulting in subtle cognitive deterioration. Findings of relatively low memory performance associated with heavy ecstasy use are highly consistent across different studies and user populations. In addition, low performance in tests of higher executive function were reported in some but not all studies. The important questions about progression, persistence, or reversibility of damage after long periods of abstinence have to be addressed in future studies with longitudinal design. PMID- 12078017 TI - [Vascular dementia]. AB - The term vascular dementia refers to dementia syndromes which are caused by hypoxic-ischaemic brain lesions. Lesions found in vascular dementia such as complete and incomplete infarctions, selective necroses, and others are nonspecific. The characteristics and severity of the clinical syndrome are determined by the size and topography of the ischaemic lesions. Among others, age and pre-existing brain atrophy are risk factors for the development of dementia based on vascular lesions. There is a high comorbidity of Alzheimer's disease and vascular dementia. It can be presumed that ischaemic lesions and Alzheimer-like pathological changes exert additive effects in the manifestation of the clinical dementia syndrome. The diagnostic process follows three steps: 1. presence of a dementia syndrome, 2. presence of cerebrovascular disease, 3. evidence for a relationship between 1 and 2. Present diagnostic criteria, such as "International Classification of Diseases" (ICD-10), "National Institute of Neurological Disorders," "Stroke-Associated Internationale pour la Reserche et l'Enseignement en Neurosciences" (NINDS-AIREN), and "Alzheimer's Disease Diagnostic and Treatment Centers" (ADDTC) describe differing constellations and show little congruence. Estimates of the prevalence depend highly on the set of criteria used. Hence, they differ considerably. PMID- 12078019 TI - [Etiologic and pathogenetic heterogeneity of Alzheimer disease]. AB - The present review discusses whether the clinically and pathologically uniform phenotype of Alzheimer's disease (AD) consists of different subtypes with respect to etiopathogenesis. In a minority of patients with AD, the disease is determined by mutations of genes on chromosomes 1, 14, and 21. The amyloid cascade hypothesis provides a conclusive model for etiopathogenesis of these cases. The majority of patients, however, suffer from the sporadic subtype of AD, the etiopathogenesis of which is presently unknown. This subtype's pathogenesis is determined by a cascade of events, among which age, a disturbance of neuronal insulin receptor function leading to a deficit in acetylcholine and available energy, and membrane instability are the most important factors. The effects of these factors are modulated by the presence or absence of apolipoprotein E4. Thus, the development of this form of AD is characterized by dynamic cellular biological changes in the brain which are independent of the metabolism of the amyloid precursor protein and genetic factors. Therefore, the sporadic subtype of AD may be regarded as an age-associated metabolic disease of the brain. PMID- 12078020 TI - [Altered perception of temporal relations in schizophrenic psychoses]. AB - Basic perceptual skills involving the central nervous system require the orderly temporal integration of internal as well as external information. Current research in schizophrenia increasingly centers on the accompanying neurocognitive deficits. In association with schizophrenic psychoses, there have been frequent reports of altered temporal processes, but explicit research on the perception of temporal relationship is still rare. Using concrete operationalized neuropsychological procedures, the present pilot study addressed the question whether schizophrenic patients (n = 27) differ from a healthy control group (n = 18) concerning their ability to judge correctly the temporal order of visual stimuli. We found a significant impairment in basal temporal perception among patients. Moderating variables such as antipsychotic medication, attention deficits, or motivation effects did not appear to be essential explanatory factors for this finding. Thus, our findings indicate a fundamental disturbance in the temporal coordination of neuronal network functions in association with schizophrenic psychoses and are in line with neurophysiological, neuroanatomical, and neuropsychological overlappings of schizophrenia and temporal perception. PMID- 12078021 TI - [Screening for neuropsychological deficits--a screening test also for dementias?]. AB - To assess further the psychometric features of neuropsychological deficit screening (NDS), three groups of patients (41 demented, 23 not demented but cognitively impaired, and 50 controls) were examined and the results compared with two older tests, the Mattis dementia rating scale (MDRS) and the mini mental status test (MMST). As a result, the discriminating ability of NDS (modified for this study) was comparable to both other tests without surpassing them in either demented or cognitively impaired groups. An abbreviated version allowed for an improvement in economy without sacrificing discriminating properties. The advantage of NDS thus lies in the diversity of functions assessed, while its screening power is comparable to the two other tests used for comparison. PMID- 12078022 TI - [Questioning or automatic recall of memorized fragments?]. AB - Considering memory loss and its interpretation using structurally dynamic methods, objections to spoken psychological memory research techniques are raised: mnesic fragments are not recalled but can be naturally called forth with a combination of practice and deactualisation. PMID- 12078023 TI - [Recall and pseudo-memory. On the yearning to be a trauma victim]. AB - Memories are not called up from "storage" but instead are constructed anew in each case. Although many experiments have proven that memories are visual and inaccurate, many psychotherapists still assume that memories which surface during therapy are realistic representations of facts. They do not take into account that reminiscences (pseudomemories) of events can be planted in the memory by the imagination or through behavioral pressure. In light of this, the question arises as to why some patients during therapy tend to invent in particular memories of traumatic early childhood experiences. The authors assume that certain suggestive elements come to bear with victimization. The advantage gained is of great importance and has many facets. The case of Wilkomirski proves that mystification of the ego via identification with victims is not only provoked in psychotherapeutic treatment but also is a means of gaining public attention and support. As concerns the therapeutic handling of actual emotional traumatization (whose pathogenic significance and long-term effects used to be underestimated), suggestive and autosuggestive processes play a large role. In this respect, modern trauma research and psychotherapy are faced with special challenges. PMID- 12078025 TI - [Cerebral toxocariasis: a rare cause of cognitive disorders. A contribution to differential dementia diagnosis]. AB - We report on a 65-year-old woman with depressive symptoms and cognitive deficits confirmed by psychometric tests. Routine blood tests, serology, EEG, and cranial computed tomography (CCT) being normal, the CSF revealed an eosinophilic reaction and a positive antibody titre against Toxocara canis. After treatment with two oral courses of albendazole, the eosinophils had disappeared, whereas the antibody titre had increased. One year later, the patient's cognitive symptoms had improved, and new antibodies against toxocara were seen in the peripheral blood. This increase in antibodies represents an expected immunological reaction to the increased exposition to toxocara antigen under effective therapy. The infection might go back to the patient's youth, when she was a shepherd for several years and in close contact to dogs. A review of the literature did not yield other reports of toxocara infections leading to cognitive or other psychiatric symptoms. Taking the toxocara infection as the cause of this patient's cognitive defects, we propose that lumbar punction becomes part of the diagnostic standard in differential diagnosis of dementia. PMID- 12078024 TI - [Microdeletion 22q11.2. A too rarely diagnosed genetic change in psychiatric illnesses]. AB - Microdeletion 22q11.2 with an estimated incidence of 1:4000 is known to cause the DiGeorge syndrome (DGS) or the velocardiofacial syndrome (VCFS), both usually being diagnosed in the newborn period or childhood. Recent studies have shown that children suffering from VCFS frequently develop psychiatric disorders in late adolescence or adulthood. Here we report the case of a 30-year-old man presenting with slight facial dysmorphisms, hypoparathyreoidism, minor cardiac anomalies, and slight cognitive impairments who had developed a severe personality disorder which eventually led to the diagnosis of microdeletion 22q11.2 with maternal inheritance. Psychiatric patients should be thoroughly examined for typical signs associated with this chromosomal anomaly. Genetic diagnosis is necessary because of the 50% probability of inheritance with possibly severe congenital anomalies. In view of a prevalence of 2% in an unselected group of patients with schizophrenic psychosis, microdeletion 22q11.2 is likely to be underdiagnosed. PMID- 12078026 TI - [Interferon-induced paranoid psychosis. Review of the literature and case report]. AB - Adverse neuropsychiatric effects under interferon treatment have often been reported. However, the pathogenetic mechanisms have not yet been fully explored. Most frequent are affective disorders, but psychotic decompensations can also occur, as reported in the following case. Thus, taking a detailed psychiatric history is required before starting a therapy with interferon, since past and current psychiatric disorders might imply an increased risk for psychiatric adverse effects. PMID- 12078027 TI - [Delusional depression as differential dementia of the Alzheimer type diagnosis]. AB - For several years, a 68-year-old patient had been treated for a dementia of Alzheimer's type and finally admitted to a closed gerontopsychiatric nursing home. According to information from his relatives, he first developed psychotic symptoms 3 years prior to admission in our psychiatric department. Several months later, he developed a lack of drive, lack of interests, and reduced emotional reaction. After a standard diagnostic work-up (psychiatric, neurological, and general medical examination, CSF examination, laboratory analyses, cranial computerized tomography), we began electroconvulsive therapy (ECT) with a working hypothesis of major depression with psychotic symptoms. The ECT was able to ameliorate rapidly the psychiatric status of the patient. After 10 weeks of inpatient treatment, he could be discharged from the hospital and was able to take up his usual professional activities on a voluntary basis. This article describes the need for an early differential diagnosis of dementias. Standardized and differential diagnoses of dementias are necessary for an antidementive therapy as well as for the detection of potentially curable diseases. PMID- 12078028 TI - [Charles Bonnet syndrome in an elderly patient with bilateral vision loss, hyperthyroidism and relative digitalis overdose]. AB - Charles Bonnet syndrome (CBS) is characterized by the presence of visual hallucinations in elderly, mentally healthy people. We report a visually impaired 90-year-old woman suddenly complaining of visual hallucinations, suffering from hyperthyroidism and a relative digitalis overdose. The diagnosis of CBS could be made after the exclusion of an intoxication and other neurological and psychiatric syndromes. In this case, visual hallucinations ceased without specific psychopharmacological therapy. A brief review of this organic hallucinosis, differential diagnosis, especially hyperthyroidism-induced psychosis, and digitoxin-induced psychosis is given and current therapeutic strategies are suggested. PMID- 12078029 TI - [Erotomania and Fregoli syndrome. A case report]. AB - A case report of a female schizophrenic patient is presented who showed a coincidence of erotomania and Fregoli syndrome with some interesting symptom constellations only rarely mentioned in the literature. Organic and psychodynamic factors were found in the etiopathogenesis of this disorder. A combination of psychotherapy and neuroleptic therapy was partly successful. PMID- 12078030 TI - [Mental incapacity and incapacity to make a decision and their status for determining legal guardianship]. AB - The juridical guidelines for legal capacity demand the distinction between continuous and temporary mental disorders. A case report illustrates the problematic nature of this distinction. In addition the parallels to legal requirements for guardianship are discussed. PMID- 12078031 TI - [Pharmacological support of exposure therapy in anxiety disorders. Animal experiment studies]. AB - The fear-potentiated startle paradigm (i.e., the enhancement of the startle response in the presence of an aversive conditioned stimulus) is a valuable cross species model for the study of the neuronal mechanisms underlying fear and anxiety as well as of the cognitive control of those aversive states. Walker et al. present data from a study in rats indicating that extinction of fear in the fear-potentiated startle paradigm is enhanced by the partial agonist of the N methyl-D-aspartate receptor D-cycloserine (DCS). The authors therefore suggest tests of DCS in clinical trials as a potential pharmacotherapeutic adjuvant for exposition therapies in fear and anxiety disorders. PMID- 12078032 TI - [Psychiatric disorders in persons with mental handicap guideline]. PMID- 12078033 TI - [Transcultural psychiatry guideline: goals and activities]. PMID- 12078034 TI - [Franz Joseph Gall (1758-1828). Comments on the title page picture]. PMID- 12078036 TI - Reddish-brown papules on the leg. Lesions are associated with edema, but not pruritus, and may have a systemic cause. PMID- 12078035 TI - Methicillin-resistant Staphylococcus aureus. Management of asymptomatic colonization and outbreaks of infection in long-term care. AB - Asymptomatic colonization with methicillin-resistant Staphylococcus aureus (MRSA) is common in long-term care facilities, but the burden of symptomatic infection appears to be low. Patients known to be MRSA carriers should not be refused admission, and routine cultures to identify carriers are not warranted. In the absence of symptomatic MRSA, no measures beyond routine infection control and standard precautions are necessary. Increased rates of infection should prompt investigation of a potential outbreak and initiation of more drastic infection control measures. During an outbreak, both infected and colonized residents should be isolated until transmission has been halted. Using short-term nasal application of mupirocin ointment for MRSA-colonized residents and staff implicated in the outbreak may help break the chain of transmission. PMID- 12078037 TI - Delirium. Pragmatic guidance for managing a common, confounding, and sometimes lethal condition. AB - Virtually any medical illness, intoxication, or medication can precipitate delirium, an acute confusional state common among older persons. Delirium is associated with a high risk of morbidity and mortality, thus management requires thorough assessment and swift but careful action. A range of nonpharmacologic interventions can aid management of delirium, but in general, emergent, empiric pharmacotherapy is indicated for acute cases. Key to assessment and diagnosis is ruling out dementia and depression, determining the presence of delirium, and establishing an underlying cause. Several screening tools are available to aid this effort. Vigilance can help reduce the high number of patients discharged with unresolved symptoms. PMID- 12078038 TI - The Charles Bonnet syndrome. Visual hallucinations caused by vision impairment. PMID- 12078039 TI - Delirium. More common than stroke; it befuddles both clinicians and seniors. PMID- 12078040 TI - [Reviews for Gram stain]. AB - The Gram stain is a rapid examination method providing useful information for diagnosis and treatment of infection. However, a molecular biology inspection and rapid immunological method have been developed, and gram stains have been disregarded recently. Therefore, I explain the usefulness of gram staining for rapid diagnosis, rapid treatment, and the economy. Moreover, I will explain gram stain method "Bartholomew & Mittwer(B&M)" which replaces the Hucker method. PMID- 12078041 TI - [Advances of laboratory testing for the rapid diagnosis of tuberculosis]. AB - Rapid diagnosis and treatment are extensively important for preventing transmission of Mycobacterium tuberculosis: In 1994, The Centers for Disease Control and Prevention, CDC, has published recommendations for the rapid diagnosis in the laboratories, in which smear result should be reported within 24 h, detection and identification within 10 to 14 days, and susceptibility within 15 to 30 days. New technologies, therefore, should be implemented in the laboratory. There have been significant advances in the practice of microbiology. Nucleic acid amplification, rapid culture system, antigen detection, and ATP assay system have provided new approaches to the rapid diagnosis of tuberculosis. In the present paper, we review studies which evaluated the reliability, rapidity, and requisite diagnostic capability for each method. Furthermore, we propose an appropriate test algorithm in the clinical laboratories. PMID- 12078042 TI - [Innovated diagnostic method for bacteremia]. PMID- 12078043 TI - [Rapid diagnosis of infectious diseases at the site of outpatient clinics]. AB - It is important to diagnose the pathogen as quickly as possible to accelerate the start of appropriate treatment of patients with infectious diseases. Recently, rapid diagnostic tests improve the accuracy of diagnosis of the pathogen. Furthermore, it is also available to do the test with very simplified method by using portable kits. Many PCR and antigen detection techniques have been developed to improve the diagnosis of specific pathogens, such as influenza virus, Legionella, Chlamydia, Mycoplasma, Helicobacter pylori and Mycobacterium tuberculosis. Especially, urinary antigen detection kits are useful at the site of outpatient clinics. PMID- 12078044 TI - [Laboratory tests for infectious diseases and medical reimbursement]. AB - Emerging and reemerging infectious diseases, drug-resistant bacteria and nosocomial infection are becoming increasingly serious social problems. To control infectious diseases, it is important to develop effective therapies and to achieve complete prevention of pathogen transmission. For these purposes, rapid detection of pathogenic microorganisms is essential. Recent development and technical innovation in laboratory tests and diagnosis for infectious diseases are remarkable. New techniques such as antigen detection and genetic diagnosis, neither of which require conventional culture, have been intensively developed and introduced into clinical practice. These methods are now widely accepted as effective diagnostic tools because of their high specificity, high sensitivity and rapidity. In this article, we describe the burden of medical expenses under the law on novel infectious diseases and medical reimbursement for diagnostic tests for these diseases. PMID- 12078045 TI - [The newly certified 105 Japanese medical technologists in clinical microbiology]. AB - Interest in quality assurance(QA) in clinical laboratories in Japan has increased over the past 30 years. We have however been lagging behind countries such as the USA, Canada and the UK in QA of clinical microbiology. The main problem of QA in Japan is human resources. There are only about 400 laboratory physicians certified by the Japanese Society of Laboratory Medicine(JSLM). Almost no academics in microbiology are interested in QA and they mostly lack clinical competence. There is a small number of faculty positions, and promotions are mostly based on research productivity while medical graduates are increasingly drawn to bench work for basic, short-term research. The Japanese Society for Clinical Microbiology (JSCM) was established in 1990 in order to promote the development of clinical microbiology and its relevant fields in Japan. And 2001 was a milestone in sustained efforts of the JSLM to initiate qualifying examinations of medical technologists(MT) in clinical microbiology. 105 MT in clinical microbiology were newly certified by the Joint Committee of JSCM, JSLM, Japanese Association of Medical Technologists (JAMT) and College of Clinical Pathology of Japan(CCPJ). The certified MTs have appropriate educational background and are well motivated. With good on-the-job training, they are expected to perform effectively various tasks, including laboratory management. Recent radical changes in the health care delivery system have also had serious implications on laboratory services and QA of microbiological tests. The primary goal of the clinical microbiology laboratory is to provide accurate diagnostic testing and high-quality service at a low cost for its customers. It is believed that the Joint Committee and the newly certified MTs will contribute to narrowing the gap between Japan and other countries in clinical microbiology. PMID- 12078046 TI - [A go-between the laboratory medicine and clinical chemistry]. AB - The Joint Annual Meeting of the Japanese Society of Laboratory Medicine and the Japanese Society of Clinical Chemistry is held in Yokohama at the first time in the history of both Societies. A chance of this time, we should consider the subjects to cooperate each Society. At first, standardization of the clinical tests at clinical laboratory field including clinical chemistry should be promoted by the both Societies. At second, EBLM(evidence based laboratory medicine) making the core of EBM (evidence based medicine) is very important tool practicing science based medicine by the reason of the getting out from experience-medicine. The hard effort which find out evidence on the clinical laboratory tests from enormous literature must be carried out on many clinical test items. In the Japanese Society of Laboratory Medicine, EBLM Committee was formed at last year and the work has been started. At third, if serum and other body fluids as well as tissues which are got from patients are used outside the purpose, ethics should be asked. Ethics Committee in both Society will make a guideline for the ethics for the handling the materials from patients including genetic subjects. At forth, clinical laboratory is said to be necessary the method of quality management(QM). The accreditation process is needed under unified national control survey, if we promote QM. At fifth, medical science rapidly advances at twenty-first century. Especially, there will be advanced at the fields of gene therapy, organ substitution, prevention and complete therapy of arteriosclerosis diseases and diabetes mellitus, terminal care, infectious diseases control and so on. The effective way of activating clinical laboratory is to related to those medical fields. PMID- 12078047 TI - [DNA analysis for the post genome-sequencing era]. AB - With the completion of the human genome sequencing, the new post genome sequencing era has started. The major subjects are clarifying the function of genes to apply this information to medical as well as various industrial fields. Various DNA analysis methods and instruments for gene expression profiling as well as genetic diversity including SNPs typing are required and have been developed. Here, the history and technologies related to DNA analysis including the Wada project in the early 1980's, and the Human genome project from 1990 are described. Various new technologies have developed in this decade. They include a capillary gel array DNA sequencer, DNA chips, bead probe arrays, a new DNA sequencing method using pyrosequencing and an efficient SNP typing method by BAMPER. PMID- 12078048 TI - [Implication of single nucleotide polymorphisms in association study: mitochondrial variations as another genetic markers for hypertension]. AB - Although multiple nuclear gene polymorphisms have been identified as potential risk factors for hypertension, the linkage between extranuclear DNA variations and hypertension has been uncertain. We investigated whether mitochondrial DNA(mtDNA) polymorphisms are implicated in Japanese hypertension. We used direct sequencing methods to search for single nucleotide polymorphisms(SNPs) in a hypervariable segment of the mitochondrial control region in each blood sample from 20 hypertensives and 20 normotensives. Then, we determined the distribution of two SNPs, T16223C and C16362T, in 183 hypertensives and 193 healthy subjects in the Aomori population in the northern area of Honshu island of Japan. The relationship between the gene polymorphism and hypertension was evaluated using chi-square test. Seventy SNPs were found there and the number of SNPs in each individual was significantly greater(p = 0.0111) in hypertensives than in normotensives. The C16223 genotype was more frequent in hypertensives than in normotensives(p = 0.0018). There was no significant difference in C16362T variant frequency between the groups. From these results, we conclude that mtDNA SNPs were enriched in Japanese hypertension and that the mtDNA C16223 genotype may be one of the genetic susceptibility factors for hypertension. PMID- 12078049 TI - [Increase of soluble Fc gamma RIIIa derived from macrophages in plasma from patients with atherosclerosis]. AB - Macrophages play a major role in the development of vascular lesions in atherogenesis. The cells express Fc gamma RIIIa(CD16) identical to that in NK cells, but with a cell type-specific glycosylation. In contrast, neutrophils express Fc gamma RIIIb in two allotypes, NA1- and NA2- Fc gamma RIIIb. These Fc gamma RIIIs are released from the cell surface on activation, and these soluble forms(sFc gamma RIII) are present in plasma. In the present study, we measured sFc gamma RIIIaM phi in plasma with Immuno-PCR with newly-developed anti-Fc gamma RIII mAb, MKGR14(mIgM), which recognizes Fc gamma RIIIaM phi specifically. In healthy donors, the level of sFc gamma RIIIaM phi increased with age. In contrast, the sFc gamma RIIIa level correlated with the number of NK cells in peripheral blood, and the level of total sFc gamma RIII(sFc gamma RIIIa plus sFc gamma RIIIb) correlated with the number of neutrophils. There was no correlation among the levels of three sFc gamma RIIIs was observed in healthy donors. The levels of these sFc gamma RIII were significantly increased in patients with coronary artery disease (CAD) compared with age-matched healthy donors. The sFc gamma RIIIaM phi level was related to the number of significant coronary artery stenoses, and correlated with the sFc gamma RIIIa level, total cholesterol, HDL cholesterol(negatively), LDL to HDL ratio, triglycerides and body mass index. These findings may show that the macrophages are activated during the incipient stage of atherosclerosis, and that sFc gamma RIIIaM phi may serve as predictive marker for atherosclerosis. PMID- 12078050 TI - [Role of newly developed technology in blood coagulation disorders]. AB - Clarification of blood coagulation disorders has been made progress based on newly developed technologies. ANTITHROMBIN III: In 1979, studies were carried out on antithrombin III(AT III) as a blood coagulation inhibitor from the first Japanese case of congenital AT III deficiency found in the laboratory sample. An alternative anticoagulant to maintain fluidity in extracorporeal circuit in an AT III deficient patient requiring hemodialysis was needed. In 1989, it was firstly introduced argatroban, a synthetic thrombin inhibitor as an alternative anticoagulant instead of human antithrombin III concentrate plus heparin regimen during hemodialysis. In 1992, success of good delivery in pregnant woman with an AT III deficiency was obtained to maintain normal level of AT III by the concentrates. DYSFIBRINOGENEMIA: Two families of congenital dysfibrinogenemia in clue to abnormally high levels of serum fibrinogen degradation products(FDP) were found. Polymerization detect due to dysfibrinogenemia induced high level of serum FDP and normal level of fibrin d-dimer products. HEPARIN COFACTOR II DEFICIENCY: A patient with congenital heparin cofactor II(HCII) deficiency was found in clue to four episodes of repeated restenosis under heparin-anticoagulated coronary angioplasty. For preventing the restenosis, argatroban as an alternative of heparin was used during coronary angioplasty. No restenosis after the angioplasty was appeared in anticoagulation with argatroban. Genetic analysis of the HCII was described that gene protein is secreted normally, but rapidly degraded in the circulation. FACTOR VII: In Japanese elderly, increase of Factor VII clotting activity was found to relate to high risk of cardiovascular disease. Elevation of activated FVII(FVIIa) as an activation marker of FVII in cardiovascular disease indicates to be an independent risk factor for cardiovascular disease. HEPARIN INDUCED THROMBOCYTOPENIA: HIT: HIT is believed to be less frequent in Japanese because of lack of recognition due to poor understanding of HIT's paradox. A survey in Japanese indicated to be no less frequent compared with that of the West. A causativity for HIT has been identified as antibodies against PF4/heparin complexes. As detection of the antibodies can easily be done by ELISA test, the test is desirable to become popular. Treatment for HIT is recommended to stop heparin and start a thrombin inhibitor. In Japan, argatroban and nafamostat mesilate are selected as an alternative agent for HIT, but both drugs have no approval for Japanese HIT patients. PMID- 12078051 TI - [Novel renal function marker, ATP--establishing the normal range, cases of anti tumor drags administration for urinary-track tumor, diabetic diseases and a newborn baby]. AB - Urinary free ATP assay by the firefly luciferin-luciferase method is a rapid and simple method for determining renal function, especially uriniferous tubule function. Normal range of urinary free ATP concentration, daily ATP excretion in urine, urinary ATP/creatinine value and ATP decomposition activity in urine is 1.1 x 10(-9)-3.4 x 10(-8) M, 4.0 x 10(-9)-4.1 x 10(-8) mole, 5.0 x 10(-13)-5.9 x 10(-11) mol/mgCr and 100-77% express for the remaining rate of additional ATP, respectively. A significant correlation was found between free ATP concentration and daily ATP excretion in urine with a correlation coefficient of 0.84. In cases of anti-tumor drug(cisplatin = cis-diamminedichloroplatinum II) administration for urinary-track tumor, abnormal urinary free ATP concentration and ATP decomposition activity in urine were clearly demonstrated after a few days of cisplatin administration. The appearance of a tendency toward abnormal relative ATP values were similar to changes in beta 2-MG and NAG values. Diabetic patients often demonstrate unusually high values of urinary free ATP concentration. In asphyxia of the newborn, urinary ATP/creatinine value were significantly higher than those in healthy newborn, but urinary NAG values did not differ. PMID- 12078052 TI - [Plasma endotoxin assay by supersensitive reagent for the diagnosis of gram negative bacteremia]. AB - With a novel supersensitive reagent, we evaluated the utility of measuring plasma endotoxin level for the rapid and sensitive diagnosis of gram-negative bacteremia. Subjects were 112 febrile(more than 38 degrees C) patients suspected of having bacterial infection and 170 samples were collected. Venous blood was obtained aseptically before administering antibiotics. Blood culture and endotoxin assays were performed simultaneously with these materials. Plasma endotoxin levels were positive in 64 samples when the cut off index was postulated at 0.35 pg/ml, while only 5 samples when the cut off index was postulated at 5 pg/ml. When the cut off index was at 0.35 pg/ml, sensitivity was 86%, while it was 14% when the cut off index was 5 pg/ml. Gram-negative rods(GNR) were detected by blood culture in 14 cases and the average period to detect GNR was 14.8 hours. The presence of circulating viable bacteria is diagnosed by blood culture, but because of the serious consequence of bacterial sepsis, treatment is initiated even in the absence of an identifiable organism. Since plasma endotoxin level can be assayed in about 2 hours, it will be more practical if we adjust the cut off index according to the clinical situation. PMID- 12078053 TI - [Serum leptin levels in patients with rheumatoid arthritis are correlated with body mass index]. AB - Leptin, a 16-kD protein of the ob gene product, is produced by adipose tissue and acts on the hypothalamus to suppress neuropeptide Y secretion which reduces the appetite. It has been demonstrated that serum leptin levels in healthy subjects are correlated with body mass index(BMI). Leptin is also produced by stimulation of inflammatory cytokines such as tumor necrosis factor(TNF)-alpha and interleukin(IL)-1. Rheumatoid arthritis(RA) is one of the chronic inflammatory diseases in which high serum cytokine levels are noted. In this study, we measured serum leptin levels(s-leptin) by RIA kit in 20 healthy subjects (10 males and 10 females) and 49 RA patients(14 males and 35 females) and the markers for joint inflammation such as ESR and CRP. There was no difference in s-leptin between controls(male: mean +/- SD = 5.6 +/- 3.0 ng/ml; female: 7.8 +/- 4.5 ng/ml) and RA patients(male: 4.9 +/- 3.2 ng/ml; female: 9.1 +/- 5.7 ng/ml). S leptin was correlated with BMI in both healthy subjects and RA patients. However, there was no correlation between s-leptin and the values of ESR or CRP, disease stages in RA patients. In conclusion, s-leptin in RA patients reflects BMI but not joint inflammation. PMID- 12078054 TI - [Development of the detection system for Mycobacterium tuberculosis DNA by using the isothermal DNA amplification method ICAN]. AB - The isothermal and chimeric primer-initiated amplification of nucleic acids(ICAN) is a new isothermal DNA amplification method composed of exo- Bca DNA polymerase, RNaseH and DNA-RNA chimeric primers. We developed the detection system, combined the ICAN with luminescence detection by a probe hybridization, for Mycobacterium tuberculosis DNA targeting the IS6110 insertion element. We examined performance tests of the system. This system was able to detect one copy of Mycobacterium tuberculosis DNA for only 3.5 hours, and performance of the system was equivalent or better to the Roche PCR system. We also examined a detection system by using magnetic beads, which system could shorten detection time for 2.5 hours. It was shown that the ICAN system was an efficient and sensitive detection system for Mycobacterium tuberculosis DNA from mass samples. PMID- 12078055 TI - Type-specific antibody for hepatitis C virus detected by use of NS-4 peptide and hepatitis C virus genome in Korea. AB - Hepatitis C virus, often possessing mutant genes, have the features that allow them to avoid host's immunologic response and further cause chronic progressive infections. Therefore, it is essential for those patients infected of HCV to receive improved diagnostic procedures. And it is equally important to investigate the course of disease progression and the response to treatment. The goal of this study is to review the efficacy of the third generation immunoblot assay and standardized RT-PCR-Hybridization assay, and in contrast with genotype identification(genotyping), the followings were briefly evaluated for the efficacy of serotype identification(serotyping) by using NS-4 peptide in the observation of the course and in the treatment of patients with HCV hepatitis. 1. The true positive rate in 132 cases showing repeated positives with 3rd generation anti-HCV EIA are 81.8% by immunoblot assay and 75.8% by RT-PCR Hybridization assay. 2. The 79.5% concordance of immunoblot and RT-PCR Hybridization assay is shown. The negative results from immunoblot assay are also negative in RT-PCR-Hybridization assay. 3. Among 95 patients with HCV hepatitis patients in 95 cases, the serotype 1, 2 and 4 were 53.2%, 45.2%, and 1.6%, respectively. In 29 cases, the genotypes of patients with HCV showed 1b in 15 cases, 2a/2c in 8 cases, 2b in 2 cases and mixed type in 4 cases. 4. In comparison between serotype and genotype, they showed 75.9% concordance. But serotyping showed higher efficacy in experimental procedures and sampling conditions, with more convenience. Based on above evaluation and reference review, it is reasonable to check with 3rd generation immunoblot assay the samples producing repeated positive results from anti-HCV EIA. For more definitive diagnosis of HCV infection, it is appropriate to confirm and double check with standardized RT-PCR-Hybridization assay. Lastly, it is strongly suggested that for observation of progression and for choice of interferon treatment, serotype identification(serotyping) is more useful in practice than genotype identification (genotyping). PMID- 12078056 TI - Gender-specific medicine the partnership for women's health at Columbia University: what's in a name? PMID- 12078057 TI - Balancing acts: support for workers to provide family care. PMID- 12078058 TI - Gender dichotomy in reactivity to the vasoactive oxidant hydrogen peroxide in spontaneously hypertensive rats. AB - OBJECTIVES: To determine: 1) whether ovariectomy increases low-density lipoprotein (LDL) cholesterol; 2) if increased LDL cholesterol correlates with increased blood pressure; and 3) whether arterial muscle reactivity to the vasoactive oxidant hydrogen peroxide (H2O2) is gender-dependent in spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY). METHODS: SHR and WKY females were ovariectomized at four weeks of age. Blood pressures were monitored weekly. Blood was collected at the time of sacrifice at 16-18 weeks, and caudal arteries were excised. LDL cholesterol was measured. Caudal arterial muscle was contracted with high-calcium egtazic acid (EGTA)/ionomycin (calcium ionophore) and relaxed with zero-calcium EGTA/ionomycin. H2O2-induced contractions were elicited in zero calcium. H2O2 reactivity was compared for SHR and WKY, male and female arterial muscle. RESULTS: Ovariectomy increased LDL cholesterol in both SHR and WKY but blood pressure increased only in SHR. Male SHR and WKY arterial muscle was more reactive to H2O2 compared with female preparations. CONCLUSION: Gender dichotomy in the development of hypertension does not appear to be due directly to the LDL-lowering effects of estrogen but is likely due, at least in part, to lower arterial reactivity to vasoactive oxidants, such as H2O2, with a consequent reduction of oxidant-induced vasoconstriction in female SHR. PMID- 12078059 TI - Characteristics and mortality of acute stroke patients: are there any gender differences? AB - OBJECTIVE: To investigate gender-related differences in the characteristics and mortality of stroke patients admitted to the hospital. METHODS: Prospective study of 296 consecutive patients admitted with a stroke and studied for epidemiologic features, risk factors, prior morbidity, neurologic details, acute-phase characteristics, acute mortality, and 90-day mortality. Patients were divided into male and female groups for analysis. RESULTS: Male-to-female ratio was 163 (55%) males to 133 (45%) females. There were no differences in mean age (P = .10), risk factors, stroke syndromes, infarct size, acute-phase characteristics, or complications. Females were found to be less frequently on prior aspirin (P = .02) and had fewer lacunar strokes (P = .005). A trend was observed for higher acute-phase mortality in females (P = .065); mortality at 90 days was similar (P = .71). CONCLUSION: There are no gender-related differences in patients admitted with acute strokes. A minor trend for higher acute-phase mortality in females may be due to their less frequent lacunar strokes and less frequent use of aspirin prior to stroke. Men and women with acute strokes should have access to similar services. PMID- 12078060 TI - Cognitive flexibility, reaction time, and attention in women with premenstrual dysphoric disorder. AB - OBJECTIVE: To evaluate cognitive functioning throughout the menstrual cycle in women who suffer from premenstrual dysphoric disorder (PDD) and controls. RESEARCH HYPOTHESIS: Measures of cognitive function will discriminate between women with prospectively documented PDD and women without PDD during the late luteal phase of the menstrual cycle. PARTICIPANTS: The participants consisted of 37 women who met operationalized, prospectively documented criteria of premenstrual dysphoric disorder and 32 women without PDD who served as controls. DESIGN: A series of neurocognitive tasks were used to assess selective, sustained attention, reaction time, and cognitive flexibility in women with PDD and controls. Tests were administered on two occasions: once during the follicular phase of the menstrual cycle, and once during the late-luteal phase. RESULTS: Repeated measures and discriminant analyses failed to demonstrate significant group differences between women with PDD and controls in neurocognitive functioning. CONCLUSIONS: The findings indicated that cognitive functioning in the late-luteal phase was not compromised in either group, in spite of subjective reports from the PDD group. The findings suggest that complaints of cognitive dysfunction may be mediated by altered perceptions and sociocultural expectations rather than by identifiable cognitive deficits. PMID- 12078062 TI - Women at greater risk for lung cancer. PMID- 12078061 TI - Gender differences in irritable bowel syndrome. AB - Irritable bowel syndrome (IBS) is a functional gastrointestinal disorder characterized by abdominal discomfort or pain associated with changes in stool frequency and/or stool form. It is the most common gastrointestinal disorder in both primary care and gastroenterology clinics. IBS is associated with high health care costs, disability, work absenteeism, and significant morbidity. In this article, we review the gender differences in epidemiology, diagnostic criteria, physiology, psychological features, and responses to therapy of IBS. PMID- 12078063 TI - Better understanding gender differences in CAD. PMID- 12078064 TI - Facility profile. Renovation brings chemotherapy to the patients. PMID- 12078066 TI - In sync. Benefits accrue through reliability-centered maintenance. AB - Reliability-centered maintenance (RCM) programs establish best practices to determine the condition of equipment as well as find root causes that signal imminent system or product failures. The reliability-centered approach monitors the conditions of systems, limiting the frequency of inspections. The reduced labor can often lead to significant bottom-line cost savings. PMID- 12078065 TI - Knowledge is power. Certification programs offer health facilities professionals the opportunity to demonstrate expertise. PMID- 12078067 TI - A star is born. Benchmarking tool helps hospitals achieve energy efficiency. PMID- 12078068 TI - Rent control. Selecting the right textile rental service. PMID- 12078069 TI - The Army goes rolling along.... PMID- 12078071 TI - When culture and medicine collide. PMID- 12078072 TI - The ADAA Foundation is ten years old. An interview with Dan Parrilli. PMID- 12078073 TI - Success with pit and fissure sealants. PMID- 12078074 TI - [Present situation of food-poisoning]. PMID- 12078076 TI - [Genetic diversity of enterohemorrhagic Escherichia coli]. AB - Enterohemorrhagic Escherichia coli(EHEC) causes diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome. Genomic comparison of an EHEC O157: H7 strain isolated from the Sakai outbreak and a benign laboratory strain K-12 revealed that acquisition of a large amount of foreign DNA has promoted the genetic diversification of E. coli strains. In the emergence of O157: H7, bacteriophages, in particular, played an important role. EHEC are a group of strains with several serotypes, each belonging to different E. coli lineages. Even in an O157 lineage, significant phenotypic and genetic heterogeneities are observed. Recent knowledge on the significance and the generating mechanism of such heterogeneity in EHEC strains are summarized. PMID- 12078075 TI - [PulseNet Japan--network system for the utilization of epidemiological information and the results of pulsed-field gel electrophoresis]. AB - Since 1996, we have been analyzing DNA pattern of enterohemorrhagic Escherichia coli(EHEC) O157: H7 isolates in Japan by the use of pulsed-field gel electrophoresis. The method, capable of discriminating genotypical difference of the isolates, enabled us to find the contaminated food such as salmon roe which was the causative agent for the multiprefectual outbreaks in Japan. These outbreaks which we are referring as diffuse outbreaks seem to be increasing in number, because it reflects that some of widely distributed or mass-produced food products are being contaminated by pathogens such as EHEC. In order to find a diffuse outbreak promptly and prevent it becoming large, we are constructing a network, called PulseNet Japan, for sharing the results of pulsed-field gel electrophoresis and epidemiological information among municipal public health institutes and National Institute of Infectious Diseases. PMID- 12078077 TI - [Development of vaccine for enterohemorrhagic Escherichia coli infection]. AB - Efforts on the development of vaccines against enterohemorrhagic Escherichia coli (EHEC) infection has been described in this review. Two kinds of vaccines were developed and these have been targeted for in humans and cattle. One vaccine candidate is toxoid, which uses an inactive form of Shiga toxin(Stx). A part of B subunit, each B or A subunit or one or two amino acid mutated holotoxin were developed as a toxoid vaccine candidate. The other candidate was bacterial surface antigen such as a live attenuated EHEC and hybrid between non-toxic LPS and toxoid. A live attenuated vaccine against EHEC O26: H11, O157: H7, O139: H1 were developed. Further a live attenuated vaccine candidate of Vibrio cholerae O1 expressing Stx1-B, Shigella flexneri expressing S. dysenteriae O-antigen and Stx1 B, or Salmonella Typhimurium expressing O111 antigen were developed. Hybrid type vaccine candidates were also developed with O111 LPS and tetanus toxoid, O157 LPS and exotoxin, and O157 LPS and Stx1-B. PMID- 12078078 TI - [Effect of cytokines on the expression of Shiga toxin toxicity]. AB - Shiga toxins(Stxs), which are produced by enterohemorrhagic Escherichia coli and Shigella dysenteriae serotype I, induce proinflammatory cytokines including tumor necrosis factor-alpha, interleukin(IL)-1 beta, IL-6, interferon-gamma, and chemokines such as IL-8 in intestinal epithelial cells, vascular endothelial cells, and monocytes/macrophages in vitro and in kidneys and spleen in vivo. Cytokines induced by Stxs and lipopolysaccharide enhance the toxicity of Stxs via up-regulation of the expression of Gb3, a Stx receptor, and infiltration of neutrophils. Stxs bind to neutrophils and transmigrate across intestinal mucosa and are transported to the target organs through bloodstreams. Stxs induce cytokines in vascular endothelial cells and peripheral blood monocytes and may injure organ tissues, finally resulting in hemolytic uremic syndrome and encephalopathia. PMID- 12078079 TI - [Retrograde transport of Shiga toxin 1 in human renal tubular epithelial cells]. AB - To displays its cytotoxicity by initiating RNA cleavage, Stx1 is required to be transported from endosomes to the endoplasmic reticulum, where the translocation of the A-subunit to the cytosol occurs. In the case of established cell lines, it is reported that a large proportion of the internalized Stx is transported to lysosomes for degradation and only a fraction of the toxin molecules can reach to the cytosol. By using primary culture cells, however, we observed that Stx1 is effectively delivered to the cytosol in normal human renal tubular epithelial cells. Investigation of retrograde transport mechanism of Stx should provide insights into the analysis of Stx-mediated cell damage and lead to improvements in therapeutic approaches for diseases caused by Stx. PMID- 12078080 TI - [Mechanism of A/E lesion formation produced by enterohemorrhagic Escherichia coli: O157--role of EspB, Tir and cortactin]. AB - Enterohemorrhagic Escherichia coli(EHEC) belongs to a family of pathogens which cause attaching and effacing(A/E) lesion on target cells. The following event is the alteration of the host intestinal cell cytoskeleton to form a pedestal-like structure. As a result of some recent breakthrough discoveries, EHEC injects effector proteins, EspB and Tir, into the host cells by type III secretion machinery and they modify cellular function and lead the signaling events by its direct binding with cytoskeletal proteins, alpha-catenin and talin. Cortactin is also accumulated to adherent site and involved in EHEC-induced A/E lesion. As a result of those interaction between bacterial effector proteins and host cytoskeletal proteins, EHEC can trigger the rearrangement of the host cell's actin cytoskeleton and induce the A/E lesion. PMID- 12078081 TI - [Indicators for early diagnosis of enterohemorrhagic Escherichia coli infection and methods for final diagnosis]. AB - Bloody diarrhea, abdominal cramp, history of barbecued beef for dinner at a restaurant and evidence of outbreak are important signs of Enterohemorrhagic Escherichia coli (EHEC) infections. Although early antibiotics therapy, such as fosfomycin and fluoroquinolones, is considered to be effective in Japan, stool cultures before antibiotics are essential for final diagnosis. In cases of hemorrhagic colitis or hemolytic uremic syndrome, when cultures are negative, detection of anti O-type specific lipopolysaccharides IgM antibodies is useful for estimate of causative EHEC. In our hospital, non-O157 EHECs increase in number as the causes of pediatric HUS since 1999. PMID- 12078082 TI - [Rapid detection methods of enterohemorrhagic E. coli O157 and interpretations of those results]. PMID- 12078083 TI - [Treatment in initial stage of VTEC infection]. AB - Verocytotoxin producing Escherichia coli(VTEC) causes gastrointestinal infections worldwide. In at most 8% of the children in Japan who are infected with VTEC, hemolytic-uremic syndrome(HUS) develops soon after the onset of diarrhea. Treatment with antibiotics does not ameliorate VTEC infections, and in some studies from western countries, it has been associated with worse clinical outcomes. It was recently indicated in Japan that early administration of fosfomycin to the patients with VTEC infection can decrease the risk of HUS. Moreover, early administration of Synsorb-Pk with high affinity to verocytotoxin to the patients with gastrointestinal VTEC infection was demonstrated to decrease the incidence of very mild and mild HUS, but it did not decrease the risk of moderate and severe HUS. In the developing stage of HUS, intravenous administration of fluid and electrolytes should be determined cautiously to prevent hyponatremia and systemic congestion. PMID- 12078085 TI - [New drugs that prevent cytotoxicity of Shiga toxins]. AB - Shiga toxin(Stx) produced by enterohemorrhagic E. coli is the virulence factor that causes not only enterohemorrhagic colitis but also fatal complications, such as hemolytic uremic syndrome. To prevent the complications, new strategies targeted to Stx have been tested, mostly using mimics of the trisaccharide structure of neutral lipid Gb3, the receptor for Stx. One group of such new drugs are agents that can bind to Stx in gastrointestinal tract and prevent its spread to extraintestinal sites, and the other group are water-soluble neutralizers that suppress Stx cytotoxicity in the circulation. Although most of these are now under the laboratory investigations, one of these drugs may hopefully be utilized clinically to prevent hemolytic uremic syndrome in future. PMID- 12078084 TI - [Therapy for children with Shiga toxin-E. coli-associated hemolytic uremic syndrome]. AB - Hemolytic uremic syndrome(HUS) is characterized by microangiopathic hemolytic anemia, thrombocytopenia, and acute nephropathy. Clinical features and outcome of children with HUS initiated by infections with Shiga toxin(Stx)-producing strains of Escherichia coli(E. coli) infection are different from those of patients with the other forms of HUS or thrombotic thrombocytopenic purpura(TTP). Childhood Stx E. coli-associated HUS usually recovers spontaneously and dose not require specific treatments including plasma therapy. In contrast, a general consensus has been achieved that plasma exchange or infusion should always be tried in adult HUS/TTP to minimize the risk of death or long-term sequelae. In this paper, we briefly reviewed therapy for patients with Stx-E. coli-associated HUS. PMID- 12078086 TI - [Norwalk virus: recent findings]. AB - Acute gastroenteritis due to Norwalk virus(NV), a member of the family Caliciviridae, is a common illness worldwide. NV is a human pathogen causing food borne and water-borne diseases that occur in various epidemiological settings. The disease is mild and self-limiting and the symptomatic phase lasts 24 to 72 hours. Genetic analyses of the polymerase and the capsid protein indicate that NV is grouped into two clusters, genogroup I and genogroup II. The development of recombinant virus-like particles made a great impact on the development of diagnostic assays such as antigen ELISA and antibody ELISA. Strong control measures are needed to prevent oyster-related outbreaks. PMID- 12078087 TI - [Molecular epidemiology of Norwalk virus]. AB - Norwalk virus is one of the most important diarrheal viruses. Although the virus was discovered in 1970, the genetic analysis has started from 1990. The virus appears in sporadic cases but also in outbreaks in children and adults. The symptoms are vomiting and diarrhea, but rarely are convulsion and others. The molecular epidemiological method is useful to detect the pathogen in environments and hosts. As the results of molecular epidemiology, it was found that the common virus with the identical sequence spreads in the world and the virus with unique sequence spreads in a limited area. In Japan, Norwalk virus infection occurs in early winter in children, while, rotavirus infection in late winter. In addition to food borne transmission, air borne transmission is strongly suggested. Norwalk virus infection is recognized to be higher prevalence following rotavirus infection. PMID- 12078088 TI - [Epidemiology on Norwalk virus-related gastroenteritis outbreaks among elderly persons living in nursing homes]. AB - Person-to-person transmission of Norwalk virus(NV) is important factor for outbreaks in nursing homes in addition to foodborne and waterbone transmission. NV spread easily among residents and staffs through the care process and highly infectious contamination generated by vomitus and feces remains on the environmental surface. As a result, the outbreak continues for long period(a few weeks) compared with foodborne gastroenteritis outbreak in restaurant. For prevention and control of NV outbreak in nursing homes, it is necessary suitable efforts to break off the transmission routes; frequent handwashing, adequate measures against infectious materials such as stool and vomitus, cleaning with an appropriate germicidal product, and to minimize any mingling of residents and visitors until termination of the outbreak. PMID- 12078090 TI - [Phylogenetic analysis of 14 strains of Norwalk-like viruses: identification of the region in the genome for genotyping]. AB - 'Norwalk-like viruses(NLV)', a member of the family Caliciviridae, are the major causative agent of acute gastroenteritis and genetically divided into two groups, geno-group I(GI) and genogroup II(GII). We have determined complete nucleotide sequences, of 9 new NLV strains. Using this information together with 5 known NLV sequences, we investigated the criteria to further classify genotypes of NLV. Validation of the topological error based on the bootstrap value and the branch length(distance) identified two potential subgenomic regions suitable for genotyping. They were the RNA dependent RNA polymerase(RdRp) and the capsid N terminal/Shell domains(capsid N/S). When the distance distribution analysis was performed, the RdRp-based classification did not separate the strains into internal clusters within genogroup. Furthermore, a diversity plot analysis of the complete nucleotide sequences of the GII Saitama U1 strain indicated that the genotype was different between the RdRp and the capsid N/S, suggesting that these strains are the genetic recombinants. Therefore, RdRp is not suitable for genotyping. On the other hand, the clustering based on the capsid N/S successfully distinguished the NLV equally to the cluster according to the antigenicity determined by both antigen and antibody ELISAs with recombinant virus-like particles. We would like to recommend to use the capsid N/S for the genotyping. PMID- 12078089 TI - [Genomic organization of Norwalk-like viruses and functions of viral gene products]. AB - Norwalk-like viruses(NLVs) that is one genus of the family Caliciviridae are major causative agents of nonbacterial acute gastroenteritis in human. NLVs have not yet been propagated in cell cultures and model animals, which restricts the fundamental studies. However, cDNA from several NLVs can be expressed in the cell free system, bacterial cells, insect cells and mammalian cells. Studies on polyprotein processing by virus-encoded protease, enzymatic properties of RNA helicase, protease and RNA polymerase, capsid assembly, interaction between viral proteins or genomes and cellular proteins, the molecular mechanism of translation and transcription, and the crystal structure of the capsid protein and other viral proteins are in progress. Results will be useful for development of drugs for diarrheal therapy. PMID- 12078091 TI - [Identification and genotyping of Norwalk virus by stringent microplate hybridization of PCR products]. AB - There are many genotypes of Norwalk virus(NV), and the microplate hybridization method for identification and genotyping of NV requires preparation of many probes. However, the method is simple and the detection sensitivity is high at 10 ng/ml DNA, and many samples can be simultaneously examined in an identification test and genotyping of NV. Increasing the hybridization temperature increases precision and the homology rate between a probe and DNA fragment can be estimated. Mixed genotypes of NV are often detected in a same sample of NV infected patients and oysters. In such cases, genotypes cannot be determined by direct sequencing. However, this method is applicable to genotyping of cases infected with mixed genotypes. Genotypes can be determined without sequencing by preparing many probes. PMID- 12078092 TI - [Rapid and efficient detection method of Norwalk virus]. AB - A rapid and efficient RT-PCR with fluorogenic probe(TaqMan-PCR) was developed for detection of Norwalk virus(NV) genomes in clinical specimens. We designed NV genogroup specific primers and fluorogenic probes in the junction of open reading frame (ORF)1 and ORF2. Eighty specimens from patients of gastroenteritis, in which NV-like particles were detected by electron microscopy, were examined by TaqMan-PCR and RT-PCR using primer sets previously reported; two sets in RdRp region of NV and one set in capsid region. TaqMan-PCR detected NV genome from 79 of 80 specimens(98.8%); this method showed the highest sensitivity of all other RT-PCR tested. Moreover, TaqMan-PCR was considered to be useful to recognize genogroup of NV correctly. PMID- 12078093 TI - [Newly developed Norwalk-like viruses detection ELISA by combination with monoclonal antibodies and hyperimmune rabbit sera]. AB - Norwalk-like viruses(NLVs) antigen detection ELISA was newly developed. It is constructed with monoclonal antibodies(MAbs) as a capture antibodies and hyperimmune rabbit sera as a detector antibodies. The two MAbs reacted broadly and specifically with genogroup I and genogroup II NLVs, respectively. Total detection time is less than 3 hours and at least 90 stool samples could be tested at one time. The concordance rate with ELISA and RT-PCR was 69% and it could be raised to 71% if capsid primer sets were used. Epidemiological study revealed that among 664 normal persons one sample(0.2%) was confirmed positive by both ELISA and RT-PCR, suggested that NLV carrier may exist in the normal person. The ELISA kit has advantages such as rapid, simple and multi samples detection, and useful for the first screening test. Also it is big tool for epidemiological surveillance of NLVs invasions. PMID- 12078094 TI - [Antibody-based diagnostic assays with recombinant virus-like particles]. AB - Virus-like particles were produced with 4 genogroup I Norwalk-like viruses(NLVs) and 7 genogroup II NLVs by a baculovirus expression system, and used to detect the antibody to NLVs by enzyme-linked immunosorbent assays(ELISA). Very little cross reactivity was observed between the genogroup I NLVs and the genogroup II NLVs when the antibody ELISA was done with hyperimmune sera. Infections by several genotypes of the NLVs were thought to occur in oyster-associated acute gastroenteritis, while an infection by a single genotype of NLVs was found in the illness occurred in a hospital. A high prevalence of antibody to 11 NLVs was observed in samples collected from healthy adults in Japan. PMID- 12078095 TI - [Characteristic of Norwalk gastroenteritis and its prevention]. AB - The application of recently developed molecular technique revealed the important role of the Norwalk virus(NV). The major role of the NV as agents of food and water-related outbreaks of gastroenteritis has now been recognized in the world. Moreover, a part of NV is also documented as important agents of gastroenteritis in childhood and infancy. A high rate of food-related viral gastroenteritis outbreaks was observed following ingestion of shellfish in Japan. Specific methods are not available for the prevention or control of NV gastroenteritis. Hand washing will be the most effective to decrease transmission within a family and a hospital. To control food-borne diseases a different strategy is necessary with virus and bacteria. PMID- 12078096 TI - [The relationship between epidemic gastroenteritis caused by Norwalk viruses (NVs) and acute gastroenteritis in children--based on the history of NVs in Japan]. AB - Norwalk viruses(NVs) are well known pathogens that can cause acute nonbacterial epidemic gastroenteritis in adults as well as acute gastroenteritis in children. In food poisoning cases due to raw oysters, NVs is considered as an important pathogen and the molecular epidemiological studies of NVs are the most suitable method to trace these routes of infections in conjunction with oyster supply mechanism. Furthermore, the prevalence of genotypes of NVs which are detected from cases of acute gastroenteritis in children may provide us with useful information for the epidemiological study of acute nonbacterial epidemic gastroenteritis, because investigations of acute nonbacterial epidemic gastroenteritis might be revealed to be only a piece of infiltration of NVs in nature. PMID- 12078097 TI - [Contamination of oyster sea farm with the Norwalk virus: mechanisms and control]. AB - The Norwalk virus(NV) is widely known as a cause of nonbacterial food poisoning, infant diarrhea, and acute gastroenteritis in the winter months between November and March. While it is strongly suspected that NV that is excreted by humans flows into coastal seawaters via rivers and wastewater treatment facilities to contaminate oysters that are grown in farms in the area, light has yet to be shed on the behavior of this virus in the natural environment. We therefore conducted a polymerase chain reaction (PCR) survey of NV levels in the aquatic environment of the oyster bed area of the Shima region in Mie Prefecture, whereupon the NV was detected in marine sediment, oysters, and mule clams even during the summer months, when food poisoning is infrequent. In order to assess their similarity to human-derived strains, the detected viruses and their human-derived counterparts were subjected to genetic analysis, whereupon some of the detected viruses were found to be remarkably similar to those that were previously detected in humans infected with NV. In the interests of examining methods for decontaminating NV contaminated oysters, we also conducted an assessment on a system of virus decontamination that focuses on seawater temperature and oyster metabolism, using Poliovirus Sabin strain. The decontamination system mentioned above was a closed loop, water circulating system, built on the same principles as those actually in use at oyster farms. Our experiment indicated that at seawater temperatures of both 10 degrees C and 20 degrees C, virus placed into the water tank was rapidly incorporated into the midgut glands of the oysters. Thereafter, when seawater irradiated with UV was circulated, the virus count in the oysters fell from 1/1,000 to 1/10,000 within 6 hours. These results indicated the utility of this system for virus decontamination, suggesting the possibility of significantly alleviating the risk of NV infection in humans by using this system to maintain the seawater temperature within the decontamination tank above a certain temperature, and to perform decontamination with an adequate water flow. PMID- 12078099 TI - [Enteropathogenicity of Providencia alcalifaciens, a newly recognized pathogen causing food-poisoning]. AB - Providencia alcalifaciens, a member of the family Enterobacteriaceae, has been thought to be a normal flora or an opportunistic pathogen. In 1996, a large outbreak (including 270 patients) of acute gastroenteritis occurred in Fukui. No known enteropathogens were detected in fecal samples of the patients, but P. alcalifaciens was detected from 7 of the 18 samples. The isolates were a clonal by pulsed-field gel electrophoresis. The specific antibody against the isolated strain was elevated in the patients' sera. The isolates showed invasiveness into Caco-2 cells and fluid-accumulation in rabbit ileal loops. These results indicate that P. alcalifaciens can be one of the causative agents of gastroenteritis or food-poisoning. PMID- 12078098 TI - [Vaccine development for Norwalk virus]. AB - The rationale for developing a vaccine generally is made on the basis of the clinical and socioeconomical significance of a specific viral disease. The disease burden for rotavirus gastroenteritis in infants is well evaluated and several candidate vaccines for rotavirus has now been developing. An increasing recognition of the clinical significance of Norwalk virus causing acute gastroenteritis in humans of all ages, foodborne outbreaks in adults, and relatively severe gastroenteritis in children indicates that an effective vaccine would be useful. This article reviews the immunity for Norwalk virus infection and the development of a candidate vaccine for Norwalk virus. PMID- 12078100 TI - Schizophrenia: the big questions. PMID- 12078101 TI - Ethics, culture and psychiatry. PMID- 12078102 TI - [Psychiatric care in Canada]. PMID- 12078103 TI - [Psychiatric care in Japan compared to that in France]. PMID- 12078104 TI - [Japanese psychiatric care viewed from overseas]. PMID- 12078105 TI - [Cross-cultural adaptation and mental disorders]. AB - Recently, it has not been rare for psychiatrists to treatment disorders and provide consultation on mental health problems for people from different cultures. The following is a brief outline of mental disorders and mental health problems of the people who immigrated in culturally different societies. In reviewing the literature, most of the reports demonstrate that the admission rate for schizophrenia is higher among immigrants than among people remaining in their original countries or among native-born people of the host countries. Furthermore the prevalence of mood disorders in community surveys is reported to be higher among immigrants than among native-born people of the host countries. In regard to gender difference of vulnerability to cross-cultural immigration, the prevalence rate for mental disorders is reported to be higher in female immigrants than male immigrants. Psychological adaptation of Japanese war orphans returned from Mainland China four decades after WWII revealed that psychological problems became exacerbated three months after resettlement in Japan and took three years to recover. Three major psychological symptoms that manifested in resettlement were obsession, somatization and depression. It is hypothesized that the acculturation process can be divided into four layers: the behavioral level, intellectual level, representative level and emotional level. The study of the acculturation process of Japanese war orphans demonstrated that it gradually proceeded from a surface level to deeper levels, namely from the behavioral level ->intellectual level-->representative level-->emotional level. The relationship of psychological adaptation and acculturation process is reported to be complicated. The acculturation process does not necessarily accompany psychological adaptation and vice versa. Impact of immigration on psychological manifestation revealed the following vicissitude: the asymptomatic phase for two months after resettlement-->hypochondriacal phase-->depressive phase-->paranoid phase. The vicissitude of psychological manifestations of the impact of immigration may be a reflection of the four layers of adaptive mechanisms (G. Vaillant). PMID- 12078106 TI - [Actual situation and social prognosis of women seeking psychiatric care at the emergency hostel of Tokyo Metropolitan Women's Counseling Center]. AB - The emergency hostel of Tokyo Metropolitan Women's Counseling Center, established in 1957, provides protection and care for about 600 Japanese or foreign women per year. The women housed there need social support for various reasons such as prostitution, poverty, somatic or mental diseases, or domestic violence (DV). We investigated the sociodemographic characteristics, psychiatric diagnoses and social prognoses of 2667 women who consulted the psychiatric clinic in the emergency hostel between 1961 and 1997. Seventy-four women consulted the psychiatric clinic per year, on average. Most were aged between 20 and 49. During the study period, there was a gradual decline in the number referred through the prostitution prevention law. Psychiatric diagnoses at the first visit varied widely. Annual comparison showed a gradual decrease in schizophrenia and manic depressive illness, but an increase in substance abuse, psychogenic reaction, and personality disorder. Sociodemographically, most subjects appeared to have been children from underprivileged backgrounds. The social prognoses of 930 cases judged in March 1998 were good in 25%, moderate in 23% and poor in 48%. The poor prognosis group contained significantly more women with schizophrenia and personality disorder than the other two groups. The poor prognosis group tended to include more cases of substance abuse, while the good prognosis group contained more cases of depression and mental retardation. Women in the DV group tended to have more children than those in the non-DV group, and to have a higher prevalence of psychogenic reaction and a lower prevalence of schizophrenia. The DV group also tended to include more subjects with a moderate social prognosis and fewer subjects with a poor social prognosis. Specialized treatment should be provided for women after emergency admission to the hostel and this treatment needs to be aimed at improving social adaptation of the hostel residents, especially those with schizophrenia, personality disorders and substance abuse. Victims of DV should receive more conscious attention. In particular, prevention of mental disorders should be aimed not only at the residents but also their children. PMID- 12078107 TI - [Investigation of recent problems in an acute care psychiatric ward considering the number psychiatrists working in the ward]. AB - In July 2001, psychiatric wards for acute treatments (PWAT) were investigated in Japan using a questionnaire to clarify current and recent problems in 79 PWAT. The questionnaires were sent to wards, patients and psychiatrists and were returned by 72.2% overall. The number of admissions per ward was calculated as 21 patients per one month, and comprised half of all admissions to the hospital. 50% were schizophrenia, 17% were affective disordes and 16% involved drug abuse. Seventeen patients were discharged from PWAT per one month, and comprised 43% of all patients discharged from the hospital. These results indicate that both 21 patients admitted and 17 patients discharged per month and needs to maintain the essential standard for PWAT and the standard should be come more flexible as admission from the other unit of ward than PWAT. As rate of re-admission within 3 months after discharge was around 10% of the total number of patients in the ward, 3 months was considered suitable length of acute treatment in the field of psychiatry in Japan. There was one psychiatrist working in PWAT, and specialized psychiatrists had 17.4 patients, the most number of patients among types of psychiatrist. Simulations of one psychiatrist to 16 and to 32 patients in PWAT were performed to determine how many psychiatrists were needed for a ward. When the ratio was 32 patients to 1 psychiatrist, it was necessary to increase the number of psychiatrists to a ward by 1, and in the case of 16 patients, 1-3 psychiatrists were needed. These indicate the standard number of psychiatrists for PWAT should be at most one psychiatrist for the ward or all of the psychiatrists working in PWAT should be allowed to work simultaneously in other wards. Preparing wards to treat acute phase psychiatric patients is a very important role of each psychiatric hospital, the standard for PWAT should include not only a high level of medical staff, but also preparing easy criteria for each hospital. PMID- 12078108 TI - Mistakes in the air: do the rules of the cockpit transfer to the ER? PMID- 12078109 TI - Air-quality standards recommended for feeding operations. PMID- 12078110 TI - Case # 1. Maxillary first molar retreatment. PMID- 12078112 TI - Case # 3. Why is my tooth still sensitive to cold? PMID- 12078111 TI - Case # 2. Mandibular incisor retreatment of silver points. PMID- 12078113 TI - Case # 4. Why is my tooth still sensitive to hot? PMID- 12078114 TI - Case # 5. Maxillary premolar retreatment. PMID- 12078115 TI - Case # 6. Mandibular molar retreatment. PMID- 12078116 TI - Case # 7. Mandibular molar retreatment. PMID- 12078117 TI - Case # 8. Post perforation repair. PMID- 12078118 TI - Case # 9. Focal sclerosing osteomyelitis. PMID- 12078119 TI - Case # 10. Perforating resorption. PMID- 12078120 TI - Case # 11. Pulpal pain vs. periapical pain. PMID- 12078121 TI - Implant treatment planning and rehabilitation of the anterior maxilla, Part 2: The role of autogenous grafts. AB - Implant-supported restorations have become and continue to be a very popular clinical option for patients who are either partially or completely edentulous. This article describes the management of a resorbed anterior maxilla using autogenous bone grafts harvested from the patient's mandibular symphysis region. Following an adequate healing period, the patient's treatment was completed by an implant-supported restoration. The case is used to illustrate surgical options available in the restoration of the anterior partially edentulous maxilla. PMID- 12078122 TI - Clinical decision-making for successful root canal treatment. PMID- 12078123 TI - [Clinical trial of YU BHK Rabivak vaccine against rabies in volunteers]. AB - The BHK (baby hamster kidney) continual cell line has been used for years in the production of rabies vaccine for animals. Becouse of the rich harvesting of rabies virus from this cell substrate, there is no need of the an additional virus concentration, so that the vaccine production can be organised in small local Pasteur Institutes as well. Although its long-range use in the veterinary medicine has proved that the BHK line is safe and that the vaccine is immunogenic, its use in the human medicine has been only recently studied. According to the latest recommendations of the Expert Committee for the Biological Standardisation of the WHO from 1998, residual cellular DNA in the vaccine is not a potential risk because the inactivation of the vaccine by the beta-propiolactone completely inactivates its possible biologic activity. In the work, the authors report the results of the third phase of the clinical trail of the YU BHK Rabivak vaccine on volunteers. The rabies vaccine studied, named YU BHK Rabivak, produced by Novi Sad Pasteur Institute, caused the generation of protective titre of virus-neutralising antibodies in all examinees: in 96.4% after 2 doses, in 99% after 3 doses, and in 100% after 4 and 5 doses of the vaccine. The registered side effects were expected, mild, did not need medical treatment, and were not more frequent than in the case of other vaccines from cell culture. PMID- 12078124 TI - Rabies: doubtful and discordant results in fluorescent antibody test. AB - By a fluorescent antibody test animal brain samples were examined for the rabies antigen. Two observers read the results. Doubtful, suspicious or weak results in the test were reanalyzed in a virus isolation test on the murine neuroblastoma cell line. From 37 samples estimated doubtful by the fluorescent antibody test, 17 were positive in the virus isolation test. PMID- 12078126 TI - [Treatment of bite wounds and cat-scratch disease]. AB - Although rabies is the most serious consequence of animal bite injuries, in urban rabies-free countries the risk of rabies is far lower than the risk of bacterial infections of the wound. The most frequent etiologic cause of the wound infections after dog bites is Pasteurella multocida, as well as in the case of bites by cats, after which cat-scratch disease may also develop, its main cause being Bartonella henselae. All bite injuries must be carefully cleaned and disinfected; it is necessary to estimate the need of antirabies and antitetanus protection, and to introduce antibiotic treatment for prevention, particularly in the case of deep sting wounds caused mostly by cats. PMID- 12078125 TI - [10 years' of production and use of human rabies immunoglobulin in Yugoslavia]. AB - Application of the rabies immunoglobuline is a compulsory part of the prophylaxis of rabies in all severe, transdermal lesions caused by rabies infected animals. Sylvatic rabies has spread in the past few years throughout the whole Yugoslavia, and human cases of rabies have also been reported in other East European countries. In order to achieve the highest level of rabies prophylaxis, apart from postinfective rabies vaccination, it is necessary to provide passive immunization using specific antibodies against rabies. After successful immunization of the young, healthy volunteers in 1990, National Blood Transfusion Institute, in cooperation with the Pasteur Institute from Novi Sad, prepared the first quantities of immunized plasma by plasmapheresis procedure and human rabies immunoglobuline. Without national production, sufficient quantities of human rabies immunoglobuline could not be provided, since the price on the world market is rather high (over $1000 per patient). PMID- 12078127 TI - Rabies surveillance and control measures in Slovenia. AB - The article briefly describes the sylvatic rabies situation in Slovenia in the period from 1980 to 2000, the reasons for the replacement of manual distribution of baits by aircraft distribution. An overview of the control measures, surveillance system and control programs is given. PMID- 12078128 TI - Clinical case of rabies in Bulgaria. AB - The history of a six-year-old girl, hospitalised in the Clinic of Infectious Diseases of the Medical University of Varna with diagnosed meningoencephalitis, obs. rabies, and epidemiological data of dog bite without adequate prophylaxis is presented. The clinical course was unusual: the symptoms of aero- and hydrophobia were not clearly demonstrated, choreic hyperkynesias, torsion seizures and high initial pleocytosis in cerebrospinal fluid were present. The pathological picture is described. Ethiological diagnosis was proven by immunofluorescence test of brain tissue. PMID- 12078129 TI - [Manifestations of visceral and ocular symptoms of toxocariasis in a 6-year-old boy]. AB - In the past 10 years we have examined 137 cases of toxocariasis, predominantly in children. Three cases were with unilateral ocular involvement. The article reviews a 6-year-old boy with left side strabismus and granulomatous chorioretinitis. Laboratory examinations revealed blood eosinophylia 24% and IgG against Toxocara canis larvae in titer 1:320 by indirect immunofluorescent assay. In epidemiologic anamnesis we concluded that about 4, 5-year-long geophagia was the source of infection. PMID- 12078130 TI - [Human rabies--Andrijevica in 1947]. AB - The highest incidence of human rabies in Yugoslavia was recorded in the post-war years, between 1946 and 1949. Medical literature also quotes the epidemic of rabies mediated by a wolf in Andrijevica in 1947. Rabies was not confirmed in the wolf by laboratory findings, but the clinical picture of wolf rabies and the clinical findings in the infected and dead people indisputably confirmed the diagnosis. The parts and organs of the animal were not adequately preserved and transported. The wolf's head reached the reference laboratory, 12 days after the rabid animal had been killed. Laboratory tests on the material from the infected or dead people were not done. The data are supplemented by new facts. Epidemiological approach was used, and interviews with the participants and the members of the injured and deceased, as well as the citizens who were thought to be able to assist in the reconstruction of the events in 1947. The protocols of Pasteur Institute and the Public Records Office of Montenegro were consulted. The author points out that the patients showed "hydrophobia, laboured breathing..." which is an undisputed symptom of rabies, a clinical confirmation of the conclusion made based on the behaviour of the infected wolf. The barking of the household dogs and actions of the herds when they were protecting their animals are probably additional reasons of high exposure to the rabid wolf. Among several available data, the author points out that there were 28 injured and 9 infected persons. This gives a death rate of 32.1%. The described rabies epidemic has had the highest number of injured and dead in Montenegro and post-war Yugoslavia. PMID- 12078131 TI - [The Pasteur Institute complex in Novi Sad as a cultural monument]. PMID- 12078132 TI - Eighty years of Pasteur Institute in Novi Sad and a century of scientific rabies prophylaxis in Serbia. PMID- 12078133 TI - [Should our daily D-vitamin intake be increased?]. PMID- 12078134 TI - [Why is the amount of amputations not reduced and the rehabilitation is halting?]. PMID- 12078135 TI - [Commitment-based out-patient care would reduce the risk of violence in schizophrenic patients]. PMID- 12078136 TI - [Neoplasms under control with the help of vascular treatment?]. PMID- 12078137 TI - [What is known about genetic factors behind chronic inflammatory bowel diseases?]. PMID- 12078138 TI - [Stem cell transplantations as a treatment of severe autoimmune diseases]. PMID- 12078139 TI - [Loss of hair]. PMID- 12078140 TI - [Prevalence of erectile dysfunctions and coital frequency]. PMID- 12078141 TI - [Sleep-disordered breathing in children with meningomyelocele]. PMID- 12078142 TI - [Diagnostics and treatment of thiamine deficiency]. PMID- 12078143 TI - [Recurrent perforations of small intestines, gastroduodenal ulcers and diarrhea in a middle-aged woman]. PMID- 12078144 TI - [Where did the pulmonary coin lesion vanish from the x-ray picture?]. PMID- 12078145 TI - [Urinary tract infections]. PMID- 12078146 TI - [RP-HPLC determination of adenosine in fermented Cordyceps]. AB - OBJECTIVE: A method for determination of adenosine in four fermented Cordyceps products by RP-HPLC is described. METHOD: C18 column was used with methanol-0.06 mol/L potassium phosphate monobasic-tetrahydrofuran (10:150:1.5) as mobile phase. The flow rate was 1 ml/min and detected at 260 nm. RESULTS: The average recovery was 98.7% and relative standard deviation was 1.10% (n = 5). CONCLUSION: This method is simple, rapid and sensitive. The paper introduces the feasibility of supersonic extraction method taking the place of reflux method. PMID- 12078147 TI - [Determination of triptophenolide in Chinese herb leigongteng by TLCS]. AB - OBJECTIVE: To establish a method for the determination of triptophenolide in Chinese herb Leigongteng (Tripterygium wilfordii Hook. f.). METHOD: TLCS was used. The sample was purified with a column and developed upwardly with CHCl3 EtOEt(2:1), sprayed with Keddle testing solution and scanned. The content was calculated with external two-point method. RESULT: The linear range was 2.1 micrograms-10.5 micrograms, r = 0.998. CONCLUSION: The method is proper for the quality control of Leigongteng. PMID- 12078148 TI - [Isozyme electrophoresis of Fritillaria puqiensis during bulb organogenesis in vitro]. AB - OBJECTIVE: Inquiring into the mechanism of bulb organogenesis of Fritillaria puqiensis. METHOD: Time course studies of the soluble protein, peroxidase and esterase were performed during the new bulb organogenesis by electrophoresis. RESULT: The atlas of electrophoresis changed greatly during the phase of differentiation and the formation of the new bulb. CONCLUSION: Different enzymes are associated with organogenesis and express activity at different times. PMID- 12078149 TI - [A study on inhibitory effect of plant growth retardants on earlier bolting of Chinese Angelica]. AB - OBJECTIVE: Seeking chemical ways of controlling the earlier bolting of Chinese Angelica (CA). METHOD: PP333, B9 and MH in different concentrations and combinations were sprayed into the CA leaves in different periods. RESULTS: In leaf proliferating period the spray could inkibit the earlier bolting, shortening the length of the stem foot, increasing the stem diameter and enhancing the leaf chlorophy 11 contents. The quality of CA production was increased as a result of greater root weight and main root volume. In leaf luxuriatino period the spray showed no marked effect. The growth of the plant would be inhibited by any spray in excessive concentrations. CONCLUSION: Leaf spray could control the earlier bolting of CA. PMID- 12078150 TI - [The comparison of component contents and pharmacological actions between two kinds of processed products of Pinellia rhizoma prepared by ginger juice and alum]. AB - OBJECTIVE: To compare the component contents in Pinellia Rhizoma processed with ginger separately on orthogonal technology and pharmaceutical technology stipulated in pharmacopoeia, and study the action of the two differently processed products on animals. METHOD: Using gravimetric method and acid dye colorimetry to determine the total alkaloids, et al. RESULT AND CONCLUSION: The experiment has shown that the contents of total alkaloids of product of Pinellia Rhizoma processed by orthogonal technology are higher than those processed by pharmaceutical technology, but the contents of digged substances of the latter are higher than those of the former, and the toxicity of decoction of the latter is also higher than that of the former. Compared with raw Pinellia Rhizoma, the two products of Pinellia Rhizoma prepared by different technologies all have shown promoting action on PGE2 contents and activity of gastric proteinase in rat gastric juice. The experimental results also have shown that on the above mentioned items. There are no obvious differences between the product of Pinellia Rhizoma processed by orthogonal technology and the product processed by pharmaceutical technology. However, the component contents of raw Pinellia Rhizoma are lower than those of the above mentioned two processed products and help inhibit PGE2 contents and activity of gastric proteinase in rat gastric juice. PMID- 12078151 TI - [Quality evaluation of EGb761, standard extracts of Gingko biloba leaves, and other Gingko biloba leave products (to be continued)]. PMID- 12078152 TI - [Chemical constituents of fruits of Gardenia sootepensis Hutch]. AB - OBJECTIVE: To demonstrate the chemical constituents of the fruits of Gardenia sootepensis. METHOD: Column chromatographic technology was employed to separate the water soluble part, and the compounds obtained were elucidated by spectral and chemical analysis. RESULT: Ten compounds have been isolated, seven of which elucidated respectively as D-mannitol, beta-sitosterol, deacetylasperulosidic acid methyl ester, geniposidic acid, geniposide, scandoside methyl ester and dimeric molecule of quinide. CONCLUSION: All the compounds were obtained from this plant for the first time. PMID- 12078153 TI - [Iridoid glycosides from Pedicula dicora Franch]. AB - OBJECTIVE: To study the components in the root of Pedicula dicora. METHODS: Column chromatography with silica gel and polyamide was employed for the isolation and purification of ingredients. The structures were elucidated by spectral method. RESULT: Three iridoid glycosides were obtained and elucidated as mussaenoside, shanzhiside methyl ester and lamalbid. CONCLUSION: All the three compounds were separated from P. dicora for the first time. PMID- 12078154 TI - [Determination of total scopoletin in Caulis Erycibes by RP-HPLC]. AB - OBJECTIVE: To accumulate data for quality control of Caulis Erycibes. METHOD: The total-scopoletin in Caulis Erycibes was determined by RP-HPLC, using CLC-ODS1 column, 0.5% glacial acetic acid in methanol-water(26:55) as mobile phase and UV detector at 310 nm. RESULT: The average recovery is 99.37% and RSD 0.39%. CONCLUSION: Determination data have been obtained for fourteen batches of three species, and the contents of scopoletin should be no less than 30 mg/100 g. PMID- 12078155 TI - [Determination of three flavonoids and ferulic acid in Pleurospermum hookeri C. B. Clarke var. thomsonii C. B. Clarke by HPLC]. AB - OBJECTIVE: To establish a reversed phase HPLC method for separation and determination of licochalcone A, ferulic acid, isoliquiritigenin and nobiletin in Pleurospermum hookeri var. thomsonii. METHOD: Using ODS column with methanol water(containing 1% acetic acid)(73:27, 28:72, 59:41) and methanol-water(65:35) as the mobile phase, the flow rate was 1 ml/min, the column temperature was 35 degrees C and UV detection at 345, 320, 345, 332 nm, different extraction methods for the sample were compared. RESULT: The average recoveries were 99.45%, 98.13%, 97.81% and 96.58%, the relative standard deviations were 0.83%, 0.99%, 0.73% and 1.55%, and the contents were 0.728, 0.124, 0.0830 and 0.0237 mg/g. CONCLUSION: This study provides a scientific basis for quality evaluation, quality control and resources development and utilization of P. hookeri var. thomsonii. PMID- 12078156 TI - [Experimental studies on antioxidative effect of tongcao and xiao-tongcao polysaccharides]. AB - OBJECTIVE: To observe the antioxidative effect of four kinds of Tongcao and Xiao Tongcao polysaccharides. METHOD: Aged mice (nine months) were abdominal administrated with four kinds of polysaccharides (Tongtuomu, Ximashanjingjiehua, Xinanxiuqiu, Ditanghua) at dosages of 160 mg/kg and 80 mg/kg for 45 days. RESULT: Four kinds of polysaccharides could obviously decrease the content of serum and liver lipoperoxide(LPO) as well as the content of lipofuscin(LF) in brain and cardiac muscle of aged mice, and also increase the activity of superoxide dismutase(SOD) of red cells in aged mice. CONCLUSION: The polysaccharide components of Tongtuomu and three kinds of Xiao-Tongcao have antioxidative effect, suggesting that these substances have antiageing function. PMID- 12078157 TI - [Effects of Squama manitis on cardiac systolic function and platelet aggregation]. AB - OBJECTIVE: To study the effect of Squama Manitis(S. M.) on cardiac function and platelet aggregation in vitro. METHOD: Isolated working rat hearts were prepared, and cardiac performance was measured before and after adding S. M. in the Krebs Hensleit(K-H) perfusion buffer. Controls with K-H buffer were also investigated. Rich platelet plasma and Poor platelet plasma were centrifuged from the blood of 15 patients with coronary heart disease. Each sample was divided into three parts for use by three groups: control, group of S. M. and group of aspirin. Platelet aggregation was stimulated by adenosine diphosphate. RESULT: LVSP, +dP/dtmax and CO of isolated working rat hearts were significantly increased after treating with S. M. liquor. S. M. had no obvious effect on -dP/dtmax and LVEDP. These parameters did not change significantly in the control. Percentages of platelet aggregation were significantly lower in the group of S. M. and the group of aspirin as compared with the control. CONCLUSION: S. M. increases cardiac systolic function, has positive inotropic effect on myocardium and inhibits platelet aggregation. PMID- 12078158 TI - [Advances in the study of glucomannan from Amorphophallus]. PMID- 12078159 TI - [Identification on crude drugs from genus Leonurus: II. Leaf and flower surface characters]. AB - OBJECTIVE: To identify the crude drugs originated from 9 species and 1 variety of genus Leonurus (called Yimucao) through leaf and flower surface characters. METHOD: Routine slide processing and the characters under microscope were observed and compared. RESULTS: Obvious characteristics for identification were revealed. CONCLUSION: The shape and length of non-glandular hairs are useful in identifying this sort of crude drugs. Leaf constants and cluster crystals are also useful as reference. However, pollen grains and secretory tissues are worthless. PMID- 12078160 TI - [Study on EEG signals data compression and spikes recognition with wavelet neural network]. AB - A novel method of EEG signals compression representation and epileptiform spikes recognition based on wavelet neural network and its algorithm is presented in this paper. Wavelet network not only can compress data effectively but also can recover original signal. In addition the characteristics of the spikes and the spike-slow rhythm are detected automatically from the time-frequency isoline of EEG signal. This method can be generalized in the field of the electrophysiological signal processing and time-frequency analyzing. PMID- 12078161 TI - [Computer assisted ENT surgery system]. AB - This paper describes a novel computer assisted ENT surgery system which is based on high-performanced 3D optical localizer and the combinational registration method for matching between pre-operative and intra-operative information. Preliminary results on cadaver specimen show the system is suited for clinical ENT surgery. PMID- 12078162 TI - [Development of new ambulatory monitor for multi-channel gastroesophageal pH combining with cardiogram]. AB - A new ambulatory monitor for multi-channel gastroesophageal pH combining with cardiogram is introduced in this paper. Key techniques of the monitor, design of hardware and software, clinical significance are described. Clinical application shows that the monitor advantages the diagnoses of functional diseases of upper digestive tract, especially is significant to distinguish the chest pain of cardiac origin and esophageal origin. PMID- 12078163 TI - [Study on the design and application of the contact Nd:YAG laser scalper]. AB - A method of designing the contact laser scalper is presented. The calculations using this method is in good agreement with the measured results in laboratory. Some operations on rabbits using the newly developed contact Nd:YAG laser scalper are performed and shown that the contact Nd:YAG laser is a promising medical equipment. PMID- 12078164 TI - [Investigation and manufacture of intellectualized tonometer bassed on the principle of linearly measuring the applanation area diameter of ocular cornea]. AB - The design principle of intellectualized tonometer based theory of linearly measuring the applanation area diameter of ocular cornea has been mainly introduced, which inclueds optics and construction design principle, principle of hardware and software design of intellectualized instrument and the calculation method of ocular pressure etc. The method and result of calibration of the tonometer are also included. PMID- 12078165 TI - [Rotating gamma stereotactic radiosurgery system]. AB - A new rotating gamma system and its clinical applications are introduced, and we compare it with stationary Gamma Knife. The results demonstrate that the former has more advantages than stationary Gamma Knife on radiation dose, processing and cost, etc. PMID- 12078166 TI - [The development of medical infusion monitoring and controlling network system]. AB - The paper provided a medical infusion monitoring and controlling network system, in which the mainframe and its extensions can realize communication with each other by means of asynchronous serial port. At the same time, every extension can test infusion speed and adjust it to the presetting infusion speed gradually. The extensions also can alarm and stop infusion automatically when infusion will be over or there is a infusion malfunction. PMID- 12078167 TI - [The development of a myocardial infarction monitoring system]. AB - In this paper, the author will discuss the system design of one portable myocardial infarction monitoring system which possesses the functions of the general arrhythmia monitoring and the alarm & emergency of acute myocardial infarction (AMI). This system also can provide accurate real time ECG signal parameters for clinic diagnosis. PMID- 12078168 TI - [Successful media exchange and network communication of CT digital image]. AB - The digital image standard in medical is discussed. Based on the DICOM standard by using the media exchange and establishing the network communication between PC and CT system, we successfully acquired the CT original digital image and convert it to DICOM standard image. The CT image can easitly be read and adjusted on the PC platform. PMID- 12078169 TI - [Microwave CT techniques]. PMID- 12078170 TI - [Expression of P27 protein in odontogenic tumors]. AB - The expression of P27 protein was investigated in 40 cases of odontogenic tumors by immunohistochemical S-P assay. The results showed that P27 expression varied in different types of odontogenic tumors. Positive signals were present in 17/20 (85.0%) of the malignant odontogenic tumors, but in 6/20 (30.3%) of the benign ones (P < 0.005). It was suggested that P27 protein had certain practical value in the differentiation diagnosis of odontogenic tumors. PMID- 12078171 TI - [Separation and purification of rat immature enamel proteins]. AB - Sprague Dawley rats were chosen to prepare immature and soft enamel tissue. The authors used a sequential dissociative extraction scheme in which the enamel matrix proteins were extracted first in guanidine HCl and then in guanidine HCl EDTA, just as the method described by Termine, et al. The first step completely extracted amelogenins and the second step got enamelin. These two groups of protein were further purified by Sephadex-G200 gel filtration chromatography and analysed by polyacrylamide gel electrophoresis. The results showed: amelogenins were resolved into three fractions eluted from the column, and expressed as A1, A2, A3, respectively. The main fraction A1, which had a molecular weight range from 18 kD to 29 kD, had a main protein band of 23 kD when estimated by SDS-PAGE. Enamelin was eluted into two fractions called E1 and E2. The main fraction E1 had a single band of 44 kD on the SDS-PAGE gel. PAGE showed that amelogenins had many subunits. The authors also observed that the amelogenins would aggregate and precipitate reversibly when pH value, ionic strength or protein concentration changed. PMID- 12078172 TI - [The degeneration and regeneration of skeletal muscle after tenotomy]. AB - In order to investigate the degeneration and regeneration of skeletal muscle fiber after tenotomy, the tenotomy were carried out on gastrocnemius muscles in 15 adult BALB/C mice. The tenotomized muscles were removed on 1, 2 and 4 weeks postoperatively. Specimens were processed for histological study with light microscope and TEM, and skeletal muscle specific protein expression analysis with myoactin (MA) and myoglobin (MG) immunohistochemical staining. The results showed that the regenerating muscle cells and the degenerating muscle fibre coexisted in tenotomized muscle. The internal structure and histochemical pattern of muscle fiber changed after tenotomy. The regenerating muscle cells fused with degenerating fibres. PMID- 12078173 TI - [A stereological study of the remaining parotid gland of rat after duct-preserved partial parotidectomy]. AB - The remaining parotid glands of rats after duct-preserved partial parotidectomy were sectioned and examined randomly by stereological methods which determine acinararea and proportional volumes (PV) of the component tissues. The volume of the remaining glands were measured at the same time. Results showed that: 1. there were no difference of the above quota 1-3 days after the operation (P > 0.05); 2. 1-2 weeks after the operation, the gland volume decreased (P < 0.05) with the PV of acini decreasing (P < 0.05); 3. 4-7 weeks after the operation, the acinar area increased (P < 0.05), the PV of acini and gland volume regained (P > 0.05). All the results indicate, with the duct preserved, the remaining parotid glands after partial parotidectomy do have the regenerating ability and significant function. PMID- 12078174 TI - [Expression of P21 and P185 in benign and malignant epithelia of cheek mucosa]. AB - To analyze the expression of P21 and P185 in normal epithelia, chronic non specific inflammatory epithelia, squamous cell carcinoma and epithelia immediately adjacent to carcinoma of cheek mucosa, immunohistochemistry technique and image analysis technique were used. The results showed: there was excellently higher expression of P21 and P185 in squamous cell carcinoma and epithelia immediately adjacent to carcinoma than in normal and inflammatory epithelia of cheek mucosa, which were compactly correlative. PMID- 12078175 TI - [Relationship between the mRNA transcriptions of bFGF and KGF in the carcinogenesis of oral mucosa]. AB - The aim of the study was to determine the expressions and effects of KGF and bFGF of FGF superfamily in oral mucosa. The mRNA transcriptional levels of bFGF and KGF in dysplasia (DYS) and squamous cell carcinoma (SCC) of buccal mucosa were observed and evaluated by in situ hybridization method. The results showed that bFGF and KGF mRNA were expressed by the mesenchymal cells in both DYS and SCC groups, but not in any of the epithelial cells. The positive reactions were located in fibroblasts, endothelial cells and some of the chronic inflammatory cells. There is an increasing mRNA transcriptional level in SCC than in DYS. The results indicated that bFGF and KGF may not only enhance the neovascularization and formation of stroma by autocrine and paracrine action, but also involved in the growth of neoplastic cells. PMID- 12078176 TI - [In vitro sensitivity of oral gram-negative bacteria to the bactericidal activity of defensins]. AB - Oral gram-negative bacteria, including Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Capnocytophaga sputigena, Provotella intermedius, Provotella melaninogenicus, have been associated with destructive periodontal infection. An ultrasensitive antibacterial assay was applied to determine the sensitivity of a battery of oral gram-negative bacteria to the bactericidal effects of the isolated human neutrophil defensins (HNP) and two kinds of rabbit defensins NP1 and NP2. All species tested were killed by HNP, NP1 and NP2 except that porphyromonas gingivalis was resistant to HNP. However, there was strain-to strain variation in sensitivity. It was concluded that the oral gram-negative bacteria were sensitive to the cidal mechanism involved in defensinmediated bacterial killing. PMID- 12078177 TI - [Microvasculature of the dog's periodontal ligament and its immediate response to orthodontic forces]. AB - The purpose of this study was to examine the microvascular architecture of dog's periodontal ligament (PDL) and its immediate response to horizontal orthodontic force. A force of 200 g was applied mesially to the mandibular premolar of a dog for 2 hours, the distal side served as the contrast side. Formalin ink was perfused into the carotid arteries bilaterally through the cannulas. After perfusion the mandible was decalcified, celloidin embedded, serially sectioned and stained with eosin. The sections were then observed by light microscope and analyzed by image analyzer. Vascular corrosion casts were prepared by the similar way with the formalin ink perfusion and examined by scanning electron microscope. The results showed: there were extensive interconnections among the PDL, dental pulp, gingiva, alveolus and the neighbour tooth PDL, which would benefit the maintenance of health and functions of the dental pulp and the periodontal tissues. Arterioles and venulae could be confirmed by the presence of characteristic differences of nuclear impressions in endothelial cell under the scanning electron microscope. The quantitative analysis showed that the tensioned PDL sites in the experimental group had a higher microvascular volume density than the corresponding sites in the contrast group, while the pressed PDL sites in the experimental group had a lower microvascular volume density than the corresponding sites in the contrast group. This should result from the hydraulic mechanism in the PDL. PMID- 12078178 TI - [Relationship between dental severe attrition and the temporomandibular glenoid morphology]. AB - This study used the indices of the temporomandibular glenoid and posterior slope of the eminence by tomography to determine the effect of severe dental attrition on temporomandibular glenoid morphology. The results showed: the temporomandibular glenoid indexes in the group with dental severe attrition and in the control group were respectively 91.49 +/- 8.20 and 92.54 +/- 8.38 (P > 0.05), and the indices of posterior slope of the eminence were 49.79 +/- 5.84 and 50.38 +/- 4.58 (P > 0.05), the difference was not statistical. It was concluded that severe dental attrition would not certainly cause abnormal temporomandibular glenoid. PMID- 12078179 TI - [The study of precedent subgingival flora colonization on four kinds of crown and bridge restorative materials]. AB - The purpose of this in vitro study was to investigate the effects of microbial colonization on different crown or fixed bridge-used restorative materials and discuss the standards for evaluating those materials microscopically. Four kinds of restorative materials such as SDA-II type medium alloy, ceramic-fused-to-metal material, heat cure composite resin for crown restoration, Plat castable ceramics and four species of precedent subgingival flora (A. viscosus, F. nucleterm, C. ochracea, S. sanguis) were used. The material specimen had been continuously anaerobic cultured with the experimental bacterial fluid for 3, 7 and 14 days respectively, then the OD values of the specimenwashing fluid which could stand for the biomass of colonized bacteria on the surfaces of materials were measured. The results indicated that the quantity of precedent subgingival flora colonization on the surfaces of different restorative materials varied among different species of bacteria, which might concern with the surface structure, composition, property of anticorrosion and antisoluability of those materials. PMID- 12078180 TI - [Maxillary expansion with magnetic force: an animal experimental study]. AB - Two adult rhesus monkeys were chosen for the experimental study of maxillary expansion with magnetic force. The duration of experiment was 70 days. The following methods were applied: 1. measuring the width between the two canines and the width between the two first molars on the casts, 2. taking occlusal radiographs for the observation of premaxillary and midpalatal sutures. 3. observing the histological sections under the microscope. The results showed the width of 3 to 3 and the width of 6 to 6 significantly increased, associating with the adaptation of periodontal tissues to the action of magnetic force. PMID- 12078181 TI - [Influence of smear layer and adhesive resin on the retentive force of amalgam restoration in vitro study]. AB - The purposes of this in vitro study were to determine whether removing of smear layer and using of self-cured adhesive can increase retentive strength of amalgam restoration and to understand the adhesive mechanism between amalgam and adhesive. Class I cavities without mechanical retention were prepared in the occlusal surfaces of 45 extracted molars which were randomly divided into 3 groups. Group 1: 10%EDTA(pH 7.4) was applied to cavity walls for 1 minute prior to amalgam condensation. Group 2: 10%EDTA was applied in the same manner as in group 1, then the walls were water-sprayed and dried. Miradapt adhesive was applied, and amalgam must be packed into cavity before the adhesive cures. CONTROL GROUP: no special treatment before amalgam condensation. The tensile strength of the restorations were measured. The adhesive interface of two samples from group 2 were evaluated by SEM and EDS. As a result, removing smear layer had little effect on retentive force of amalgam (P > 0.05), while removing smear layer and applying adhesive improved the retentive strength of amalgam fillings significantly (P < 0.05), and there were micromechanical interlock between amalgam and adhesive according to SEM and EDS observation. PMID- 12078182 TI - [In situ hybridization with digoxigenin-labelling oligonucleotide: an practical approach to skeletal tissues]. AB - The purpose of this study was to investigate the in situ hybridization technique of paraffin-embedded skeletal tissues by non-radioactive labelling oligonucleotide, Five six-week-old male SD rats were selected and the proximal epiphyseal growth plates of bilateral tibiae were removed to be decalcified in 0.5 mol/L EDTA. The tissues were embedded by paraffin and the sections were treated and hybridized with RNase-free reagents. Control groups were set to testified the sensitivity and the specificity of the experiment. The results showed that this method preserved the integrate structures and effectively detected the mRNA of the tissues. These indicated that it was an efficient and practical approach to the biomolecular research of skeletal tissue. PMID- 12078183 TI - [The use of orthognathic surgery in the treatment of maxillofacial fractures]. AB - 35 cases with bony malunion following maxillofacial fractures had been treated successfully by orthognathic surgery technique. Mini-titanium plate had been used in the rigid internal fixation. The article introduced the surgical method and discussed some primary principals which should be paid attention during operation. PMID- 12078184 TI - [Distraction osteogenesis for the correction of severe unilateral posterior lockbite: preliminary report]. AB - The aim of this study was to promote an effective approach, which was surgically assisted buccal mandibular corticotomy in the lockbite side to rapidly expand mandibular arch to correct unilateral posterior lockbite using the principle of the distraction osteogenesis. The typical subject was a 16-year-old patient with unilateral posterior lockbite. The orthodontic treatment had been done before the surgery. After the surgery, reciprocal elastic traction between upper and lower arch were used immediately in the lockbite side. The results demonstrated that unilateral posterior lockbite could be effectively corrected with unilateral buccal mandibular corticotomy and rapid mandibular expansion, and the stability appeared satisfactory. This article also discussed the indications of the approach and processes of treatment. PMID- 12078185 TI - [Experimental study of radiation effects on the mandibular microvasculature of the guinea-pigs]. AB - In the experiment, both 180 kV X-ray and cobalt-60 radiation were delivered to right mandibles of 128 guinea-pigs with different dose (1000 cGy x 4, 800 cGy x 4, 600 cGy x 4, 400 cGy x 4 within 1 week). Dental extractions were conducted 1, 3, 5 months after radiation and the irradiated animals were examined by using microvascular corrosion castings of mandibles. It was found that the number of capillaries reduced and microvascular networks broke off after radiation with small veins shrinking subsequently while the alterations of arteries were slight. This result indicated that capillary was the most vulnerable part to the damage of radiation in the vascular system. In addition the defects of vascular networks around the non-healing tooth extraction sites were also discovered with irregularly capillary proliferation in the periphery of the lesion. The phenomena suggested that microcirculation failure induced by trauma-repairing process lead to occurRence of osteoradionecrosis of jaws. PMID- 12078186 TI - [Clinical treatment observation of tea pigment for oral submucous fibrosis]. AB - 39 patients with oral submucous fibrosis (OSF) were divided into control and experimental groups. The control group included 22 OSF patients who were treated by oral administration of VitAD, VitBco and VitE, and the experimental group included 17 OSF patients who were treated with vitamins and tea pigment after their examinations of hemorheology. The result showed that 7 of 12 patients in the experimental group with abnormal hemorheology had average 7.9 mm improvement on the open degree (58.3%), and the open degree of other five patients (5/17) whose hemorheology was normal only increased 2 mm (20%). The therapeutic results of the experimental group (58.3%) were significantly better than that of the control group (13.6%) (P < 0.005). The results of this study indicated that tea pigment may possibly become a better therapy for the OSF patients with abnormal hemorheology. PMID- 12078187 TI - [The influence of porcelain-fused-to-metal crown margin to the aesthetics of gingiva]. AB - The gingival tissue of 76 anterior teeth which were hurt by margin of porcelain fused-to-metal (PFM) crowns were analyzed and the reasons of the gingival hurt and the precaution methods were put forward. The unsuccessful PFM crowns were replaced with new crowns and 88.16% of them are better than before. As the result, the gingival margin health of anterior teeth is the important factor which keep the esthetics of teeth, and the right clinical process is crucial to maintain the natural aesthetics of gingival margin. PMID- 12078188 TI - [An epidemiologic retrospective study of 1693 maxillofacial injuries]. AB - A 10-year review of 1693 maxillofacial injuries was presented with respect to age, sex, cause of injury, pattern of fracture, and associated systemic injuries. The majority of fractures in maxillofacial region were found in males, the most prevalent age ranged from 21 to 40 years. The leading cause of facial trauma was traffic accidents. Mandible fracture outranked zygomatic and maxillary fracture. Multiple fractures were more common than single fracture. The average number of fractures for each patient was 1.7. Accompanied injuries to brain and skull happened in almost half patients, which were more often than other part of the body. There was a higher incidence that midfacial fractures coincided with brain and skull damage. PMID- 12078189 TI - [Quantitative analysis on platelets cytosolic free calcium concentration and ulcerous tissue calcium in patients with recurrent aphthous ulceration]. AB - To study the relationship between the abnormal behavior of calcium (Ca2+) and the pathogenesis of recurrent aphthous ulceration (RAU) by measuring quantitatively the concentration of platelets' cytosolic free calcium ions ([Ca2+]i) and tissue calcium (Wca2+), the patients with RAU (n = 50) and the healthy volunteers (n = 30) were chosen randomly. Then platelets [Ca2+]i was determined in calcium fluorescent indicator Fura-2/AM loaded washed platelets by spectrofluorophotometer and tissue Wca2+ was measured quantitatively by OCPC autoanalyzer method. The results showed that the concentration levels of platelets [Ca2+]i in patients with RAU in attack stage and attack-free stage were significantly higher than that of the control group (P < 0.01, P < 0.05), and the concentration level of the ulcerous tissue Wca2+ was also obviously higher than that of the normal control group (P < 0.001). There was a significantly positive correlation between the platelets [Ca2+]i and the ulcerous tissue Wca2+ (P < 0.01). The results suggested that the influx disorder and overload of Ca2+ might play an important role in the pathogenesis of RAU. PMID- 12078190 TI - [Factors analysis on ceramic crack of metal-ceramic restorations]. AB - This article enumerated many possible causes of ceramic crack of metal-ceramic restorations and analysed the possible causes of ceramic crack of metal-ceramic restorations. And authors also discussed the effective ways to eliminate the improper factors to keep the good bond strength of ceramics to metal alloy and to avoid ceramic crack of metal-ceramic restorations. PMID- 12078191 TI - [The effects of TGF-beta on the proliferation and differentiation of human osteoblast-like cells]. AB - Osteoblast-like cells were isolated from embryonic human calvarium with modified enzyme digestion method. TGF-beta which is believed having biological effects on bone cells was added into the culture medium with separate concentrations. The incorporation of 3H-TdR, 3H-Proline of osteoblast-like cells and alkaline phosphatase(ALP) content of the cells were tested to study the effects of the TGF beta on the proliferation of osteoblasts. The results showed that TGF-beta increased ALP activity and the synthesis of collagen, and the cell proliferation was inhibited. It affected the proliferation and differentiation of osteoblast like cells with the best 1 ng/ml among four concentration of 0.01-10 ng/ml. These findings suggested that function of TGF-beta might be modified by other local growth factors. PMID- 12078192 TI - [The effects of IGF-II on the proliferation and differentiation of human osteoblast-like cells]. AB - Biological regulatory mechanism which supports bone cell development within the context of proliferation/differentiation relationship serves as a viable basis for addressing skeletal disorders and provides experiment and theoretic basis for selection and utilization of growth factors to reconstruct bone defect. Osteoblast-like cells isolated from human calvarium were used in this study. IGF II were investigated by adding in the culture medium with different concentration. The results showed that IGF-II stimulated DNA, collagen synthesis and ALP activity of human osteoblast-like cells. Its effects were strong for proliferation and differentiation of osteoblast cells, when 10 ng/ml of four different concentrations (0.1, 1, 10 and 100 ng/ml) were used. IGF-II was proved to be important factor in bone regeneration. PMID- 12078193 TI - [Study on the adhesion of an endemic strain of Streptococcus mutans serotype C to acquired pellicle. I. A comparison of the methods to extract adhesins from Streptococcus mutans]. AB - The adhesins of S. mutans serotype c endemic strain WD9463A were extracted from culture superant by cold ethol precipitation, or were extracted from cell surface by 0.5 mol/L phosphate buffer, 6 mol/L urea or 2% SDS solution. The composition and adhesive activities of the extracts were compared by SDS-PAGE and bacterial adhesion inhibition experiments. The results showed that the protein composition of the four extracts were similar, and all extracts inhibited the adhesion of the endemic strain dramatically. It indicates that four methods can be used to extract adhesins of S. mutans, but extracting adhesins from culture superant is better than the others. PMID- 12078194 TI - [Study on the adhesion of an endemic strain of Streptococcus mutans serotype C to acquired pellicle. II. Isolation and extraction of adhesins of an endemic strain of Streptococcus mutans serotype C]. AB - Surface proteins of S. mutans WD9463 A(c) were separated by DEAE-Sephadex A25 Chromatography and Sepharose C1-6B Chromatography. The adhesins were distinguished by bacterial adhesin inhibition experiment and were identified by PAGE, SDS-PAGE, IEP-PAGE, immunol diffusion test, GTF activity test and annalization of the sorts of amino acid. The results showed P1, two proteins with molecular weight of 117 kD and 127 kD and two proteins with GTF activity could inhibit the adhesion effectively. On the other hand, another kind of GTF could improve the adhesion of the strain. These indicated that S. mutans may be many kinds of adhesins. PMID- 12078195 TI - [An experimental study of expediting the early attachment of human gingival epithelial cells to titanium with laminin]. AB - Using in vitro culture model, the attachment of human gingival epithelial cells to laminin-coated pure titanium was studied. The average numbers of attached cells on different surface of titanium with or without laminin were calculated under catoptric fluorescent microscope and were statistically analyzed with factorial design. The results showed that Laminin could expedite the early attachment of human gingival epithelial cells to titanium. PMID- 12078196 TI - [Histologic study for effects of ox collagen in forming of new bone with quantitative method in animal models]. AB - In this study, collagen was implanted into mandibular defects of 10 dogs. After operations, the samples were examined with microscope and quantitative method at 10 periods. Results showed that collagen did not affect bone growth and new bone grew into collagen directly. There was no fibrous capsule between collagen and new bone. Eventually, majority of collagen was resorbed and replaced by bone tissue. It was concluded that collagen has a good histocompatibility. PMID- 12078197 TI - [Cellular compatibility study of ox collagen with fibroblast cells]. AB - The cell culture method was used to evaluate the cytotoxicity of ox collagen for clinical use. Direct method was chosen to examine the shape and growth of cells. Results showed that the cell shape was normal and the growth was well. The cytotoxicity of ox collagen conformed to the standards made by International Organization for Standardization (ISO). This study demonstrated that ox collagen doesn't restrain the growth of cells and has good biocompatibility. PMID- 12078198 TI - [The mechanisms of antioxidant against inflammatory lesion in animal oral mucosa. II. A study of glutathione peroxidase]. AB - In order to understand the mechanisms of superoxide dismutase (SOD) against inflammatory lesion in hamster cheek pouch, glutathione peroxidase (GPx) activity in the cheek pouch mucosa was assayed in different time. The efficacy of GPx scavenging lipid peroxide was also measured. In the SOD group GPx activity decreased significantly in the early stage (8 h). But the level of lipid peroxide in the SOD group not increase at the same time. The results of the model group were contrary. It was implied that exigenous SOD scavenged lipid peroxide in the oral mucosa sufficiently in stead of GPx. PMID- 12078199 TI - [Field trials on the efficacy of albendazole composite against intestinal nematodiasis]. AB - AIM: To study the anthelmintic effect of albendazole composite (containing 67 mg of albendazole and 83.3 mg base of pyrantel pamoate per tablet). METHODS: A randomized controlled study was carried out to compare the efficacy of a single dose of 3 or 2 tablets of albendazole composite versus a single dose of 400 mg of albendazole or 10 mg base/kg of pyrantel pamoate for treatment of intestinal nematodiasis including 1,864 cases infected with hookworm, 1,568 cases infected with Ascaris, 1,785 cases infected with Tricuris trichiura and 373 children infected with Enterobius vermicularis. RESULTS: In adults, the egg negative conversion rate of a single dose of 3 or 2 tablets of albendazole composite reached 65.0% and 52.7% for hookworm infection (P < 0.01), 100% and 100% for Ascaris infection, and 26.5% and 19.2% for Trichuris infection (P < 0.01), respectively. There were significantly better effect against hookworm with 3 tablets of albendazole composite than that with single albendazole or pyrantel pamoate (65.0% vs. 47.6% and 38.5%, P < 0.01). The effect of 2 tablets of albendazole composite against hookworm was also higher than that of single pyrantel pamoate (P < 0.01) and equal to single albendazole but the anthelmintic effect against Trichuris infection was lower than that of single albendazole (19.2% vs. 26.5%). In 2-6-year-old children, the effect of 1.5 tablets of albendazole composite against Enterobius vermicularis infection showed an egg negative conversion rate of 100% which was higher than that of single pyrantel pamoate (100% vs. 83.0%, P < 0.01). The worm collection data showed that the worm expelling action of albendazole composite was much more rapid than that of albendazole. There were no adverse effects of albendazole composite on blood picture, liver or renal function and ECG. The side effect of both 3 and 2 tablets of albendazole composite was mild and transcient. CONCLUSION: Albendazole composite exhibits a synergistic effect of both albendazole and pyrantel pamoate. PMID- 12078200 TI - [Study on detection of malaria parasite DNA by PCR-ELISA]. AB - AIM: To present a new malaria diagnostic method based on detection of malaria parasite DNA by PCR-ELISA. METHODS: According to the conserved sequence of Plasmodium SSUrRNA genes reported, a pair of primers in which one primer was biotinylated and another was unbiotinylated, suitable for DNA amplification of both falciparum and vivax malaria parasites were designed and synthesized. After denaturation and washing, the incorporated biotinylated product with avidin coated on plates previously was hybridized with the fluorescein-labelled oligonucleotide probes specific for Plasmodium falciparum or Plasmodium vivax. The color developed after adding POD conjugated with antibody to fluorescein and substrate can be semi-quantitated spectrophotometrically. RESULTS: The thresholds of parasite density for the detection of Plasmodium falciparum and Plasmodium vivax by this test were shown to be as low as 4 and 10 parasites per microliter of blood, respectively and no cross reaction was seen in the detection of falciparum and vivax malaria parasites. CONCLUSION: With promising sensitivity and specificity, this test can be used in malaria survey. PMID- 12078201 TI - [Establishment of Hymenolepis diminuta-animal model and morphology of cysticercoid]. AB - AIM: To observe the morphological changes in the process of the development of Hymenolepis diminuta. METHODS: The life cycle of Hymenolepis diminuta was established between Rattus domesticus albus and Triboliun castaneum. The morphology of cysticercoid were observed microscopically, and the ultrastructure of the body surface of cysticercoid were observed by scanning electron microscopy. RESULTS: Three phases including the mature stage, the blister stage and the protective outer membrane-forming stage during the growing course of cysticercoid were observed microscopically. Under scanning electron microscope, lots of sieve-like micropores on the surface of mature cysticercoid were seen in the second week after infection. The blister phase was found in the third week and the outer membrane measuring about 45 microns in thickness were found surrounding the cysticercoid and vesicular surface, forming a smooth cyst wall in the fourth week. CONCLUSION: The life cycle of Hymenolepis diminuta has been established in the animal-model. The finding of the three phases during the growing course of cysticercoid is reported for the first time. PMID- 12078202 TI - [Genetic analysis of deltamethrin resistance in Culex pipiens pallens]. AB - AIM: To analyse the genetic character of deltamethrin resistant laboratory strain (resistant factor being 518) of Cx. pipiens pallens. METHODS: Genetic studies were made of deltamethrin-susceptible and-resistant strains of Cx. pipiens pallens with LC-P curve constructed from the response of fourth instar larvae to deltamethrin treatment, and the resistance mechanisms were analysed with synergetic tests in PB, DEF and TPP. RESULTS: The resistant factors of F1 generation from cross and reciprocal cross are respectively 154.8 and 159.2; the LC-P curve of F1 generation gets near to that of resistant parents, D = 0.625; there exist no obvious platform in 50% mortality of BC generation and 25% and 75% mortality of F2 generation, and there are larger differences between the determining curve and theoretical curve in both BC and F2 generations, P < 0.01; the synergetic ratios of PB, DEF and TPP are 27.13, 1.22 and 1.05. CONCLUSION: Deltamethrin resistance of Cx. pipiens pallens is controlled by two or more autosomal genes, and the main gene involved is incompletely dominant; multi functional oxidase is mainly responsible for the resistance. PMID- 12078204 TI - [Estimation of vector prevalence by using pool sampling]. AB - AIM: To investigate theoretically the feasibility of the pool sampling method in estimating the vector prevalence. METHODS: The probability distribution of infected individuals in a given pool was analyzed using binomial distribution, based upon the upper limit of predicted vector infectivity. The maximal pool size was determined in case that the probability of only one infected individual within this positive pool is larger than a predetermined significance level, in this way infection rate is estimated by the number of positive pools and the judgment of predicted prevalence was addressed by the threshold of positive pool ratio. RESULTS: In comparison with the pool sampling based on the positive pool ratio the method suggested in the present paper poses a variance and precision of estimates independent on pool sizes although it requires a predicted prevalence to determine the maximal pool size. In situations of limited samples and a high prevalence this pooling method has higher estimation efficiency. CONCLUSION: Theoretically the estimation of vector prevalence using pooling methods is feasible. The estimation based on positive pool ratio is suitable for situations of lacking epidemiological data and rough precision estimation, whereas the method based on the maximum pool size provides more accurate estimates and needs a prior predicted infection rate. PMID- 12078203 TI - Effect of artemether on hexokinase, glucose phosphate isomerase and phosphofructokinase of Schistosoma japonicum harbored in mice. AB - AIM: To study the effect of artemether (Art) on hexokinase (HK), glucose phosphate isomerase (GPI) and phosphofructokinase (PFK) of Schistosoma japonicum. METHODS: Mice infected with schistosome cercariae for 4-5 wk were treated ig with Art 100 or 300 mg.kg-1 and killed 24 h or 48 h after medication for collection of schistosomes. The activities of HK, GPI and PFK of the worms were determined by measuring the formation of NADPH or consumption of NADH. RESULTS: Worms from the infected mice treated ig with Art at a single dose of 300 mg.kg-1 the inhibition rates of HK activity of female and male worms were 33.7% and 13.7%, respectively 24 h after administration. Similar results were seen in GPI activity, but 48 h after medication, the inhibition rate of GPI increased to 46.2% (female) and 32.9% (male), respectively. Worms from mice treated with Art 100 or 300 mg.kg-1, the inhibitory effect on PFK in female worms was found much higher than that of male worms the inhibition rates of PFK were 64.9%-71.0% in female worms and 16.3% 54.2% in male worms, respectively at 24 h and 48 h after treatment. CONCLUSION: The results suggest that in the glycolytic pathway of schistosome PFK might be one of the targets attacked by Art. PMID- 12078205 TI - [Comparative observation on the efficacy of several test/probe systems for detecting schistosome circulating antigen]. AB - AIM: To compare the efficacy of 5 test/probe systems for the detecting of schistosomal circulation antigen (SCA) in sera from infected rabbits. METHODS: Nine rabbits were infected with monosexual cercariae (either male or female), 7 rabbits were infected with bisexual cercariae as controls. Blood samples were collected periodically post infection (p.i). 15 rabbits were infected each with 250 bisexual cercaria and divided into 3 groups, 2 groups were treated with allyl thiourea (295-590 mg/kg) from 19 d or 46 d respectively, to inhibit their egg formation. One untreated group was used as control. Blood samples were collected weekly until 8 wk post infection. SCA detecting methods include (1) dot ELISA/McAb anti-surface membrane antigen of adult worms (8SE4), (2) dot ELISA/McAb anti-CCA (3D10), (3) Sandwich dot-ELISA/McAb anti-egg antigen (MG2), (4) Sandwich-ELISA/McAb anti-egg antigen (2H10), (5) Sandwich-ELISA/McAb anti-CAA (1B10). RESULTS: Using method (1) all of the 18 rabbits infected with monosexual cercariae were negative. Using method (2) only 1 rabbit harboured 133 male worms showed positive. Using method (5) no SCA were detected in sera from female cercariae-infected rabbits but 3 out of 9 male cercariae-infected rabbits showed positive reaction. SCA detecting results from rabbits treated with allyl thiourea: both method (1) and (3) showed negative in rabbits treated from 19 d (p.i.) but all gave positive reaction in 6-7 wk in rabbits treated from 46 d (p.i.). There were all negative detected by using method (4) and all positive at 6 wk (p.i.) detected by method (5) in rabbits of 3 groups whether treated or non treated. CONCLUSION: Various test/probe systems have different efficacy in detecting SCA but (5) of the methods (1)(2) tests/probe systems could not detect SCA in rabbits with single female cercariae infection. From the egg formation inhibition study, the SCA detected by McAb 8SE4 dot-ELISA might be related to egg maturation. Whether the surface membrane antigen of adult worm has common antigenic component with egg antigen remains to be studied. PMID- 12078206 TI - [Observation on the factors influencing infection rate of Meriones unguiculatus with Brugia malayi]. AB - AIM: To explore the factors influencing the infection rate of M. unguiculatus with B. malayi. METHODS: The survival rate, infection rate and infection intensity were observed in M. unguiculatus infected with the infective larvae of B. malayi contained in normal saline, antibiotic, glucose or culture medium RPMI1640, respectively. RESULTS: The survival rate, infection rate and rate of infection intensity in normal saline group, antibiotic group, glucose group and RPMI1640 group were 80.9%, 60.5%, and 43.4%; 98.8%, 52.9% and 30.6%; 98.0%, 68.8% and 50.0%; 91.2%, 67.7% and 51.6%, respectively. The rate and the infection intensity were higher in the jirds infected with the 6th generation of Zhejiang strain than that with the 31st generation of Guizhou strain. CONCLUSION: The antibiotic was found to raise the living rate of the infected jirds, but reduce the infection rate and infection degree intensity; whereas glucose and culture medium RPMI1640 could raise the positive rate and the infectino. The jird was more susceptible to the 6th generation of Zhejiang strain than the 31st generation of Guizhou strain of B. malayi. PMID- 12078207 TI - [Preparation and characterization of monoclonal antibodies against Clonorchis sinensis]. AB - AIM: To develop and identify monoclonal antibody (McAb) against adult Clonorchis sinensis. METHODS: BALB/c mice were immunized with soluble antigen of adult Clonorchis sinensis, the spleen cells of immunized mice were fused with myeloma cell line SP2/0. Then the hybridoma cell lines secreting high titer of McAbs were screened. The Ig subtypes and titer of the McAbs were assayed with ELISA. The antigens recognized by the McAbs were examined with IFAT and identified by immunoblot analysis. The cross reactions between the McAbs and antigens of Schistosoma japonicum, Paragonimus westermani and Cysticercus cellulosae were detected. RESULTS: 5 hybridoma cell lines secreting high titer of McAbs against adult Clonorchis sinensis were established. All five clones of McAbs belonged to IgM subtype. The antigens recognized by the McAbs were located at the intestinal tract wall of adult Clonorchis sinensis. The McAbs had no cross reactions with antigens of Schistosoma japonicum, Paragonimus and Cysticercus cellulosae. CONCLUSION: 5 hybridoma cell lines against adult Clonorchis sinensis secreting high titer of McAbs with high specificity were established. PMID- 12078208 TI - A new species of Anopheles hyrcanus group from Nei Mongol Autonomous Region (Diptera:Culicidae). AB - Morphology of Anopheles (Anopheles) hailarensis, a new species belonging to the Anopheles hyrcanus group, collected from Hailaer, Nei Mongol Autonomous Region, P. R. of China, is described in adult, pupal, larval and egg stages. It is closely related to Anopheles sinensis Wiedemann, 1828 and Anopheles heiheensis Ma, 1981. Comparisons on adult morphology among above three species and egg morphology between the new species and Anopheles sinensis are made. PMID- 12078209 TI - [Studies on extensive application of bromoacetamide in various types of schistosomiasis endemic area]. AB - AIM: To provide scientific basis for the popularization and application of molluscide bromoacetamide. METHODS: Large scale application of bromoacetamide in twenty eight counties of four provinces was investigated and its molluscicidal effect in marshland and influence on marshland grass (Cyperus) and fishes were observed. RESULTS: High molluscicidal effect of bromoacetamide was found against both snails and snail eggs in various environments. According to the results observed in different fields, the applicable dosage of bromoacetamide is 1.5-2 g/m2 for spraying method, 1.5-2 g/m3 for immersing method and 1.5-2 g/m for immersing in combination with shovelling the turf along the bank. When it was applied in fish-raising ponds or rivers at an applicable dosage, no toxic effect on fishes was found. The spraying dosage of bromoacetamide did not affect the rice seedling but it did some damage to the leaf-tips of marshland grass, however, the grass did turn green and grow. CONCLUSION: Bromoacetamide has proved to be a safe and effective mulluscicde against Oncomelania under various field conditions. PMID- 12078210 TI - [Experimental study on detection of Schistosoma japonicum 5D gene by using PCR]. AB - AIM: To search for an effective method to diagnose the schistosomiasis and evaluate the effect of treatment. METHODS: A PCR protocol was designed to detect the Schistosoma japonicum 5D gene encoding an immunogenic miracidial antigen in cercariae, eggs and adult worms. RESULTS: Specific amplification of a 262 bp DNA fragment was achieved by using PCR and a 165 bp DNA fragment by using nest-PCR in cercariae, adult worms and eggs of Schistosoma japonicum. After 40 cycles by PCR, the product could be detected by agarose gel electrophoresis. This PCR protocol was able to detect as few as a single cercariae, a single egg or a pinpoint-sized tissue of adult S. japonicum. Nest-PCR and oligonucleotide probe hybridization could further increase the sensitivity. No specific PCR product was obtained with DNA from human leucocyte, Escherichia coli, Bacillus proteus vulgaris, Streptococcus faecalis and Salmonella typhi. CONCLUSION: The sensitivity and specificity of this PCR protocol are satisfactory. PMID- 12078212 TI - Phenol red method for measuring the pH of the gut contents in Lutzomyia longipalpis (Psychodidae:Diptera). AB - AIM: To develop a method to estimate the pH of the gut contents of female sandflies using a microcapillary feeding technique. METHODS: Female Lutzomyia longipalpis were fed with a small quantity of phenol red solution (indicator range pH 6.8-8.4) before and after a bloodmeal. The colour patterns of the gut contents were recorded by video microscopy immediately after the alimentary canal was dissected out of the sandfly body, and used to determine the pH level. RESULTS: In unfed flies the thoracic mid-gut (TMG) is normally neutral, with the pH ranging between 7.0 to 7.3; and the abdominal mid-gut(AMG) is mildly alkaline from pH 7.1 to 8.4 with the maximum pH observed at the junction with the hind gut. The presence of sugar in the crop reduced the pH of the TMG to 6.8, and the presence of a recently ingested bloodmeal raised the pH of the TMG to 7.4. However, as bloodmeal digestion proceeded the pH of the TMG was reduced to acidic levels, pH 6.8 or below. CONCLUSION: The new method could be integrated with the investigation of metacyclogenesis of Leishmania parasites in vivo. PMID- 12078211 TI - [Researches on immunogenicity identification of recombinant Sj22.6(rSj22.6) kDa antigen gene of schistosoma japonicum]. AB - AIM: To identify the immunogenicity and the potentiality of rSj22.6 as a candidate vaccine antigen for schistosomiasis japonica. METHODS: rSj22.6 was tested by Western blot with specific antibodies to identify its immunogenicity. The rSj22.6 protein was isolated and purified by SDS-PAGE and injected twice into rabbits to raise antirSj22.6 serum that was tested by Western blot to recognize the native protein of adult S. japonicum and SEA. C57 mice were immunized with rSj22.6 and challenged with S. japonicum cercariae to identify the protective immunity raised by rSj22.6. RESULTS: rSj22.6 could be recognized by specific antibodies and could stimulate the rabbits to produce high level of anti-rSj22.6 antibodies. Anti-rSj22.6 serum could recognize an adult native protein with the same molecular weight as rSj22.6. The preliminary immune-challenge test resulted in significant reduction in adult worm burden. CONCLUSION: rSj22.6 possesses immunogenicity in stimulating rabbits to produce high level of specific antibody and has protective role as a candidate vaccine antigen against S. japonicum infection. PMID- 12078214 TI - [Experimental infection of Schistosoma japonicum and selection of animals]. PMID- 12078213 TI - [Chemotherapy of parasitic diseases: current status and new advance]. PMID- 12078215 TI - [Studies on cathepsins in Schistosoma japonicum]. AB - AIM: To identify the cathepsins in adult worms of Schistosoma japonicum and their proteolytic activities. METHODS: The vomitus from adult worms of S. japonicum was collected. The proteolytic activity of cathepsins in the vomitus was identified and measured using synthetic substrates with arginyl bonds. RESULTS: Specific synthetic subtracts Z-Arg-Arg-AMC (for cathepsin B) and Z-Phe-Arg-AMC (for cathepsin B and L) were degraded by the vomitus. An inhibitor, Z-Phe-PheCHN2 for cathepsin L, inhibited partially the activity of the vomitus to degrade Z-Phe-Arg AMC. The optimal pH for cathepsin B/L is 5.0-5.5. CONCLUSION: The vomitus of S. japonicum has cathepsin B- and L-like activity. PMID- 12078216 TI - [Researches on the nucleotide sequencing of the recombinant Sj 22.6 kDa antigen gene of Schistosoma japonicum]. AB - AIM: To sequence the cloned gene in pGSj24 and to identify the encoded protein. METHODS: The cloned gene in pGSj24 was digested from the recombinant plasmid by EcoRI, ligated into the M13mp19 vector and sequenced by automatic sequencer. The sequence was analyzed by Goldkey DNA and Protein Analytical Program and DNASIS Program. RESULTS: The pGSj24 cloned gene was demonstrated to be 840 bp containing one opened reading frame (ORF) with an initiation codon ATG at position 23 nt and a termination codon TAA at position 596 nt, encoding a protein with a molecular weight of 22.6 kDa. At the upstream and downstream of the ORF there were termination codons, so the encoded protein was unable to be larger. However, there was a termination codon TAA at position 11 nt, suggesting why the 22.6 kDa protein expressed separately. The nucleotide sequence of the pGSj24 cloned gene shared 95% identity with that of the corresponding part of S. japonicum 22.6 kDa antigen gene, and 99.7% identity in the encoding part. The deduced amino acid sequence analysis showed a sequence motif known as EF-Hand calcium binding domain, several endoplasmic reticulum targeting sequences and microbodies C terminal targeting signals. The possible antigen determinants were predicted within the amino acid fragments of 29-32aa, 63-68aa and 87-101aa. CONCLUSIONS: The cloned gene in pGSj24 is the gene that encodes Sj22.6 kDa antigen. PMID- 12078218 TI - [Histological changes of Cysticercus cellulosae under the action of proteinase of Omphalia lapidescens in vitro]. AB - AIM: For the purpose of finding an anthelmintic with high efficacy, low toxicity and low cost, especially from the Chinese traditional medicines. METHODS: Forty Cysticercus cellulosae were separately incubated with proteinase from artificially fermented Omphalia lapidescens for 2 h, 4 h and 8 h, and then examined for histological changes by light microscopy and compared with those of proteinase from natural dry Omphalia lapidescens after similar treatment. RESULTS: Cysticercus cellulosae were morphologically and structurally impaired after exposure to the action of proteinase of Omphalia lapidescens, and the degree of impairment was proportionally paralleled to the duration of treatment. CONCLUSION: The anti-cysticercus effect of proteinase of Omphalia lapidescens and the homogeneity of proteinases from artificially fermented and natural Omphalia lapidescens were confirmed. PMID- 12078217 TI - [Determination of polyamines in pyronaridine-sensitive and -resistant Plasmodium berghei-infected erythrocytes]. AB - AIM: To understand the relationship between polyamine metabolism and pyronaridine resistance of malaria parasites. METHODS: Kunming strain mice weighing 18 g-22 g were inoculated intraperitoneally with Plasmodium berghei ANKA strain (pyronaridine-sensitive, PS), and P. berghei ANKA strain low and high pyronaridine resistant lines (PRA and PRB), respectively. Seven days after inoculation, blood samples were collected and WBC in the blood were removed by the method of chromatography using SE-cellulose and Sephadex G25 (1:2). After thin layer chromatography, the contents of three polyamines, i.e. putrescine(PTC), spermidine (SPD), and spermine(SPM) in normal and infected erythrocytes were determined with fluorescence spectrometer. The changes of polyamine level in PS and PRB infected erythrocytes of mice treated by a single oral dose of PND 5 and 10 mg/kg respectively were also observed. RESULTS: The contents of PTC, SPD and SPM in PS infected erythrocytes were 47.5 +/- 28.6, 97.7 +/- 34.0 and 81.7 +/- 23.8 nmol/10(9) RBC (n = 9), respectively, being 1.5, 3.5, and 6.6 times of those appeared in the normal RBC (32.0 +/- 9.9, 28.2 +/- 12.3, and 12.4 +/- 15.0 nmol/10(9) RBC, n = 9), respectively. The differences between two groups were significant. Interestingly, when mice infected with pyronaridine resistant line PRA and PRB, the contents of PTC, SPD and SPM in the infected erythrocytes increased to 187.7 +/- 139.9, 258.3 +/- 137.9 and 142.8 +/- 143.8 nmol/10(9) RBC (n = 18), and 345.5 +/- 223.0, 435.8 +/- 265.4 and 189.4 +/- 171.7 noml/10(9) RBC, respectively. Except for SPM, PTC and SPD in the PRA and PRB infected erythrocytes were significantly higher than those in the PS and PRA infected erythrocytes, respectively, being 3.9, 2.6 and 1.8, 1.7 times of those appeared in PS and PRA infected erythrocytes. In PND treated group, SPD and SPM contents of PS infected erythrocytes were decreased by 31.6% and 47.3% compared with non-treated group, but the polyamine level of PRB infected erythrocytes did not significantly change after PND administration. CONCLUSION: The resistance of P. berghei to PND is related to the alteration of polyamine metabolism in the malarial parasites. PMID- 12078219 TI - [Observation on the susceptibility of Anopheles sinensis to periodic Brugia malayi microfilariae]. AB - AIM: To explore the susceptibility of Anopheles sinensis to Brugia malayi microfilariae of different densities. METHODS: The survival rate of mosquitoes, the rate of microfilariae penetrating through midgut, the mature rate of larvae into infective larvae (L3) and the mean number of L3 in the survived mosquitoes were observed. RESULTS: With the microfilariae densities of 32.5 mf/microliter and 141.5 mf/microliter, the rate of larvae matured into L3 was 36.2% and 8.7%, respectively, the mean number of L3 recovered from infected mosquitoes was 8.24 and 0.30, respectively. CONCLUSION: An apparently higher susceptibility of An. sinensis to B. malayi was found by using B. malayi microfilariae density of 32.5 66.4 mf/microliter. PMID- 12078220 TI - [Chronobiological studies on effect of cyclosporin A (CsA) against adult male worms of Schistosoma mansoni in vitro]. AB - AIM: To study the action mode of cyclosporin A (CsA) against Schistosoma mansoni in vitro. METHODS: MF1 mice were infected with Schistosoma mansoni cercariae for 6 weeks when the adult worms were recovered by portal perfusion. The male worms of S. mansoni recovered were exposed to varying concentrations of CsA at 8, 16, and 24 h in vitro. Drug induced damage to the male worm surface was chrono biologically observed throughout these experiments by SEM. RESULTS: After the male worms of S. mansoni were incubated with 1 microgram/ml CsA for 8-24 h, the tegument showed swelling of ridges with appearance of holes on their surface and detachment of a part of spines. The above damage of the tegument became more evident in male worms after incubation with 10, 15, 20 micrograms/ml CsA for 8-24 h. Moreover, incubation of male worms with 25 micrograms/ml CsA for 8-24 h resulted in significant deformation and disruption of tegument, rupture of ridges and detachment of spines. The tegumental damage of male worms of S. mansoni was dose- and time-dependent. CONCLUSION: The antischistosomal action of CsA is direct, the schistosome tegument appears to be the main site for CsA attack. PMID- 12078221 TI - [Morphological observation on Blastocystis hominis]. AB - AIM: To observe the morphology of Blastocystis hominis. METHODS: The morphology of Blastocystis hominis found in the fecal specimens from school children and diarrheal cases was examined by using normal saline smear, iodine stain and iron hematoxylin staining method in Ali, Xizang Autonomous Region and Xinjiang Autonomous Region. RESULTS: The morphology of Blastocystis hominis was found to include four types, namely vacuolized, granular, amoeboid and binary fission. CONCLUSION: The vacuole type was the most often seen and accounted for 84% of all 4 types. Blastocystis hominis infection was often found to be associated with Entamoeba coli and other intestinal protozoa, leading to diarrhea in these patients. PMID- 12078222 TI - [Experimental studies on development of Paragonimus heterotremus in rats]. AB - AIM: To study the polymorphism of the development of Paragonimus heterotremus in rats and to ascertain the host nature of the rat. METHODS: SD and Wistar rats were each infected orally with 100 metacercariae of P. heterotremus. The distribution and development of the worms recovered from rats 14-156 days after infection were observed. RESULTS: Thirty days after infection, reproductive organs appeared in the worms recovered from the body cavities. Seventy days after infection, worm-cysts were found in the lungs, livers and pleural wall, the worms in the cysts matured but the worms detected in the muscles were still stunted on the 156 th day after infection. CONCLUSION: Rat is the definitive host and paratenic host of P. heterotremus. The development of P. heterotremus is polymorphic. PMID- 12078223 TI - [Comparative observation on experimental infection with Echinococcus multilocularis in Cricetulus Migratorius and Meriones meridianus]. AB - AIM: To search for an animal model with higher susceptibility to Echinococcus multilocularis and shorter experimental period. METHODS: Cricetulus migratorius, Meriones meridianus and NIH mice were experimentally infected with protoscolices of E. multilocularis and were autopsed at different times after their infection. The development of E. multilocularis in these three species of rodents was observed and compared. RESULTS: The average wet cyst weight and the ratio of cyst weight to body weight on the 91st day after infection were 15.0 +/- 2.1 g and 32.3% for C. migratorius, 3.3 +/- 1.1 g and 6.9% for M. meridianus, 0.6 +/- 0.6 g and 2.4% for NIH mice; and the corresponding figures on the 188th day after infection were 11.8 +/- 2.7 g and 21.7% for C. migratorius, 8.1 +/- 5.1 g and 15.4% for M. meridianus and 8.4 +/- 8.3 g and 26.7% for NIH mice, respectively. On the 91st day, matured protoscolices were found in C. migratorius and M. meridianus, whereas NIH mice had matured protoscolices on the 188th day after infection. CONCLUSION: C. migratorius has higher sensitivity to E. multilocularis infection than the other two species of animals as manifested by high infection rate, well-developed vesicles and rapid growth speed. C. migratorius proved to be an ideal experimental animal for E. multilocularis. PMID- 12078224 TI - [DNA immunization and its application in the field of parasitology]. PMID- 12078225 TI - [Polymorphism of circumsporozoite protein and merozoite surface antigen of Plasmodium vivax]. PMID- 12078226 TI - [Progress in the research on the detection of serum circulating immunologic substances in schistosomiasis]. PMID- 12078228 TI - An introduction of a nationwide sampling survey on schistosomiasis in China. AB - AIM: In order to know the present status of the endemicity of schistosomiasis and to evaluate the control effect during 1990-1995, a second nationwide sampling survey on schistosomiasis japonica was undertaken during September and November of 1995. METHODS: A stratifying and clustering random sampling technique was used in the survey. Serological test, circumoval precipitin test or indirect hemagglutination assay were used for preliminary screening. Stool examination using hatching technique and Kato-Katz technique was applied for identification of the infected persons and intensity of infection and hatching test only for survey of domestic animals. RESULTS: In 195 sampled villages from endemic areas, a total of 224,819 people were examined. The infection rate of residents was 4.89% and the infection rate of farm cattle was 9.06%. In the 169 sampled villages from endemic areas under control, only 17 out of 18,302 children were identified as infected by stool examination from Hubei and Sichuan Provinces. From the results of the survey, the number of infected persons, and the number of infected farm cattle and water buffaloes were estimated to be 865,084 and 100,251, respectively, in the whole country. CONCLUSION: As compared with the data in 1989, a reduction of 47.2% in infected persons and about 50% in infected farm cattle and water buffaloes, respectively, was shown. PMID- 12078227 TI - [Longitudinal surveillance of sensitivity of Plasmodium falciparum to pyronaridine in south Yunnan]. AB - AIM: To understand the changes in the sensitivity of Plasmodium falciparum to pyronaridine in the south of Yunnan Province. METHODS: WHO standard in vivo and in vitro microtest was used. RESULTS: In 1984-1985 and 1995, 36 and 39 cases of acute falciparum malaria were observed, the average defervescence time being 32.7 +/- 16.0 h and 56.2 +/- 27.4 h, the mean asexual parasite clearance time, 64.2 +/ 22.9 h and 55.3 +/- 11.8 h, and the recrudescence rate of patients followed up for 28 days, 15.2%(5/33) and 37.5%(9/24), respectively. The resistance rates of resistant isolates by in vitro microtest were 0(0/24), 8.8%(3/34), 13.8%(4/29) and 36.4%(4/11) in 1988, 1990, 1992 and 1995, respectively, while the corresponding ID50 were 13.0, 20.0, 32.0 and 40.0 nmol/L, ID95 were 48.0, 136.0, 325.0 and 330.0 nmol/L, the mean concentrations for complete inhibition of schizont formation were 32.6, 99.5, 148.2 and 190.1 nmol/L, respectively. CONCLUSION: The sensitivity of Plasmodium falciparum to pyronaridine was found to have a tendency of continuous dropping in south Yunnan. PMID- 12078229 TI - [Comparison of the morphology of wing spot between Anopheles minimus and Anopheles fluviatilis]. AB - AIM: To compare the morphology of wing spot between Anophelesminimus Theobald, 1901 and Anopheles fluviatilis James, 1902 and find out the reliable diagnostic characteristics. METHODS: Anopheles minimus were collected from Jinuo Village, Jinghong County, Yunnan Province. Anopheles fluviatilis were collected from Lingyun County, Guangxi Zhuang Autonomous Region. Measurements of the wing-length and the length of pale and dark spots on the costa and V1 were made with a microscope fitted with an ocular micrometer. RESULTS: A total of 52 females and 60 males of Anopheles minimus and 40 females and 60 males of Anopheles fluviatilis were examined. It was found that the presence of presecutor pale spot and the ratio of sector pale spot to sector dark spot were the important diagnostic morphological characteristics between the above mentioned two species. CONCLUSION: Obvious differentiations exist in the morphology of wing spots between Anopheles minimus and Anopheles fluviatilis. PMID- 12078231 TI - [A new method for evaluating malaria surveillance effectiveness--establishment and application of an index system]. AB - AIM: To establish a comprehensive index system which is suitable for evaluating the effectiveness of malaria surveillance schemes. METHODS: Delphi method was used to screen the existed effectiveness evaluation indices by experts to determine key indices and weighted coefficients. RESULTS: Three indices, i.e. autochthonous incidence, ratio of cases detected (ROCD) and ratio of introduced/imported cases were selected. Their weighted coefficients were 0.36, 0.31 and 0.33, respectively. An index system named the malaria surveillance effectiveness index (MSEI) was built up and formulated as [formula: see text] which was applied to study cost-effectiveness of the two malaria surveillance schemes (A and B). According to the formula, the MSEIs in the study areas using the schemes A and B were 48.56 and 45.93, respectively. However, the cost for increasing a score of MSEI by implementing the scheme A was four times as high as that by implementing the scheme B. CONCLUSION: The MSEI is practicable for evaluating the overall effectiveness of malaria surveillance schemes, especially in the cost-effectiveness analysis and health-economic evaluation. PMID- 12078230 TI - [Diagnosis of Falciparum malaria by immunochromatographic test]. AB - AIM: To evaluate the applicability of rapid immunochromatographic test(ICT) for diagnosing falciparum malaria in outpatient clinics in endemic area. METHODS: With thick blood smear method as control, ICT was used for the detection of P. falciparum. RESULTS: The sensitivity and specificity of ICT in detecting P. falciparum was 94.7% and 90.3%, respectively. No cross-reaction with P. vivax was found (P > 0.05). CONCLUSION: ICT is much more rapid and simple than thick blood smear method for the diagnosis of falciparum malaria, and can be applied in the outpatient clinics in endemic area. PMID- 12078232 TI - [Malaria situation in the People's Republic of China in 1997]. AB - According to the case reporting system, the number of malaria cases in China numbered 34,645 (Taiwan Province and Hong Kong not included) in 1997, with a mean incidence of 2.91 per a hundred thousand and lethal cases of 46. However, the estimated number of actual malaria cases based on investigations at provincial level was 220,000-250,000, showing a slight diminution as compared with that in 1996. Yunnan and Hainan were continued to be the major provinces endemic for malaria, where the number of malaria cases amounted to 46% of the total in the whole country. In Yunnan Province, an increase of outbreak spots and resurgence of malaria incidence in border region were reported, while the incidence was stable or slightly declined in most other provinces, but was fluctuated in provinces distributed with Anopheles anthropophagus. Blood examination on febrile patients for 9,209,102 man/time in 1,294 counties of 18 provinces/municipalities/autonomous regions (P/M/A) with a population of 569,703,000 detected 26,816 cases positive for malaria parasite, showing a mean positive rate of 0.29%; of which the number of falciparum malaria cases were 4,114 (including mixed infections with vivax malaria, the same below), accounting for 15.3% of the total parasite positives; of the three quartan malaria cases detected, one was infected via blood transfusion. Totally 568,783 target population and migratory people were surveyed, 2,861 were positive for malaria parasite, with a mean parasite rate of 0.50%; and 586 falciparum malaria cases were among the positives. As a whole, the endemicity of falciparum malaria was still confined to 44 counties of Hainan and Yunnan Provinces. Imported falciparum cases were reported in 74 counties located in 17 P/M/A, however, no introduced falciparum malaria cases were recorded. PMID- 12078233 TI - [Sensitivity and specificity of nested polymerase chain reaction in detecting Plasmodium vivax in infected mosquitoes]. AB - AIM: To study the sensitivity and specificity of nested polymerase chain reaction for detection of Plasmodium vivax in infected mosquitoes. METHODS: Two pairs of primers specific to small subunit ribosomal DNA of P. vivax were used to amplify the specific SSUrDNA 121 bp fragment of P. vivax for detecting P. vivax-infected mosquitoes with nested PCR. RESULTS: Nested PCR could detect as few as 3 sporozoites in one mosquito or 1 infected mosquito mixed with a group of 99 normal ones. In contrast, no such specific 121 bp DNA band was detected in P. falciparum, P. cynomolgi, P. yoelii yoelii infected samples, nor in normal mosquito. CONCLUSION: The nested PCR technique we established showed high sensitivity and specificity. PMID- 12078235 TI - [Differentiation of cryptic species A and D of Anopheles dirus complex by polymerase chain reaction]. AB - AIM: To distinguish cryptic species A and D of Anopheles dirus complex using polymerase chain reaction (PCR). METHODS: A diagnostic PCR assay of species was developed by use of three primers, one derived from highly conservative 5.8 S coding sequences and two from different interspecies sequence in the second internal transcribed spacer (ITS2) of ribosomal DNA. RESULTS: Using the PCR method, specific fragments were amplified in both species, the size of fragments is 374 bp for species A and 663 bp for species D. Thirty samples of species A from AFRIMS and HN laboratory colony and seven samples of the species D from Yunnan Province were correctly identified by PCR. Satisfactory results were obtained from the amount of DNA as little as 1/1,600 of extracted DNA of a single mosquito or 1/5 of DNA derived from one leg of a mosquito triturated in water. A total of 148 field-collected specimens of Anopheles dirus from Heping(HP), Baisha(BS), Loukui(LK), and Maoyang (MY) in Hainan Province revealed fragment characteristic of species A, while 30 specimens from Mengla (ML) in Yunnan Province showed the specific fragment of species D. CONCLUSION: A simple and reliable method was developed to identify cryptic species A and D of Anopheles dirus complex and it was further verified that Anopheles dirus from Hainan and Yunnan Provinces is the species A and the species D, respectively. PMID- 12078234 TI - [Application of immunochromatographic test for diagnosis and surveillance of bancroftian filariasis]. AB - AIM: To evaluate the usefulness of immunochromatographic test (ICT) for rapid diagnosis and surveillance of Wuchereria bancrofti infection. METHODS: The monoclonal antibody based-ICT assay was used to detect filarial antigens in the sera from bancroftian filariasis patients. RESULTS: 111 out of 116 bancroftian microfilaria cases showed positive ICT reaction, giving a positive rate of 95.7%. Serum samples from 12 malayian microfilaremia cases, 33 ascariasis cases, 20 schistosomiasis japonica cases and 6 trichinellosis cases were all found negative in ICT, demonstrating a specificity of 100%. In 73 chronic filariasis cases with elephantiasis, hydrocoele, and chyluria cases, 18 showed ICT positive, among whom 16 revealed microfilaremia, suggesting that the ICT positive cases were with active infections. Of the 30 microfilaremia cases before single dose ivermectin treatment, 29 were ICT positive (96.7%). A negative conversion of microfilaremia was observed in all cases 8-14 days post-treatment. Upon following up to 6 and 12 months, microfilaremia reappeared in 5 and 7 cases, respectively. 5 out of 13 ICT positive cases were found microfilaremic at 6 month post-treatment, while among the rest of 8, 5 were found microfilaremic at 12 month post-treatment. Parallel use of ICT with routine blood film examination in Houda village, Zhecheng County, Henan Province for a post-control surveillance survey in 132 local individuals revealed 9 microfilaremia cases were also ICT positive. Of the 123 persons with negative blood examination, only 1 was ICT positive. CONCLUSION: ICT is a rapid and simple method with high sensitivity and specificity, and might be used for the diagnosis, efficacy evaluation as well as a tool for post-control surveillance of bancroftian filarasis. PMID- 12078236 TI - Immunoscreening of Schistosoma japonicum egg cDNA library. AB - AIM: To screen and identify genes encoding vaccine candidates and molecules with diagnostic potential from S. japonicum (S. j) egg cDNA library. METHODS: S. j egg cDNA library was screened by sera from SEA-immunized rabbits. Positive clones were identified after three rounds of screening and were excised automatically in vivo with helper phage. Products of PCR were used to detect insert sizes by agarose gel electrophoresis. DNA samples were obtained by purification using mini prep kit. Automatic sequencing was carried out and the sequence data were analysed using GCG software. RESULTS: Twelve clones were identified as positive and the inserts were around 1.2 kb in size. Compared with DNA sequences in GenBank, eight clones were homologous to a 70 kDa S. j heatshock protein(S. j HsP70). C18 exhibited 97.8% homology with S. j23. C22 and C23 shared 82.4% and 89.5% homology with S. j calreticulin, respectively. C12 was homologous to S. mansoni(S. m.) immunophilin. CONCLUSION: This paper first reported the results of immunoscreening S. j egg cDNA library by anti-SEA sera. Twelve positive clones were found to be related to protective immunity and/or immunodiagnosis of schistosomiasis. PMID- 12078237 TI - [Analysis of molecular karyotype of Leishmania isolates from hilly foci and plain foci of China]. AB - AIM: To analyse the molecular karyotypes of Leishmania isolates from hilly and plain foci of China. METHODS: Pulsed field gel electrophoresis(PFGE) was performed in 1% agarose at 6 V/cm in 0.5 x TBE at 14 degrees C with a pulse time of 60 s for 15 h and a pulse time 90 s for 9 h. RESULTS: Each isolate was resolved into 15 different EtBr-staining bands ranging in size from 200 to 2,200 kb, in which the karyotypes of 6 isolates from hilly foci were similar (including karyotypes of canine isolates similar with those of human isolates), and they were partly similar with those of L. infantum. The karyotypes of the two isolates from plain foci were similar, but the isolates from the two topographically different foci were different from each other. CONCLUSION: Homology exists between L. d. isolates from hilly foci and plain foci respectively, and exists in part between L. d. isolates from hilly foci and L. infantum; while heterogeneity exists between L. d. isolates from hilly foci and plain foci. The reservoir host, domestic dog, plays an important role as the infection source of leishmaniasis in the hilly foci of China. PMID- 12078238 TI - [Study on invasibility and multiplication of Toxoplasma strains with different virulence toward Vero-cells]. AB - AIM: To investigate the pathogenic mechanisms of Toxoplasma strains with different virulence. METHODS: Tachyzoites from 3 strains, Viz. RH, B36 and Fukaya strains, were challenged to in vitro cultivated Vero-cells. Systematic examinations on the earliest invasion time, the invasion rate and intracellular multiplication were performed under different cultivation conditions. RESULTS: The tachyzoites of all the 3 strains invaded the host cells within a short period after inoculation. Invasion rates were all increased along with the prolonged duration of infection. The intracellular multiplication was found to be most active in RH strain, moderate in B36, and comparatively slow in Fukaya strains. Using purified tachyzoites freed from host debris and proteins and adding sufficient FCS in the medium may facilitate the invasion and subsequent multiplication of the parasite. CONCLUSION: Strain differences in pathogenicity to the host may be correlated to the genetically predetermined multiplication capabilities of the parasites after being invaded to the host cell, and that environmental factors may give certain impact on the invasibility of the parasite. PMID- 12078240 TI - [A survey of Echinococcus granulosus infection in Camelus bactrianus in north Xinjiang]. AB - AIM: To determine the situation and characteristics of Echinococcus granulosus infection in camels (Camelus bacterianus) in our country. METHODS: The camels slaughtered in abattoir were examined for visceral infection of larval cestode of Echinococcus. The number and diameter of cysts were recorded and the content of cysts were examined microscopically to detect the protoscoleces. RESULTS: 185 out of 375 camels were positive, the infection rate was 49.3%. The metacestodes parasitized mainly in the liver and lung, the ratio between liver and lung involved was 1:0.64. The fertile cyst rate was 39.2%, the fertile cyst-carrying rate of camels was 34.8%, the cyst index was 7.53. The cyst wall was relatively thin and the cysts located singly on the surface of liver. There was no daughter cyst seen in fertile cysts. The average diameter of fertile cysts was 5.6 +/- 2.65 cm in liver and 4.83 +/- 2.03 cm in lung. The percentage of animals harboring calcified cysts was 64.3%. CONCLUSION: The infection rate of metacestodes of Echinococcus granulosus in Bactrian camel was very high in north Xinjiang and the characteristics of infection differ in some aspects from the local sheep and cattle and from dromedaries in Africa as well. Further study is needed to elucidate the significance of the infection of E. granulosus in Bactrian camels. PMID- 12078239 TI - [Accumulation and effluxion of chloroquine in chloroquine-sensitive and chloroquine-resistant Plasmodium berghei]. AB - AIM: To explore whether the difference in the amount of chloroquine (CQ) accumulated in CQ-resistant (CR) and in CQ-sensitive (CS) P. berghei is involved in the infected erythrocytes (RBC) or in the parasites. METHODS: The concentration of CQ in infected RBC and parasites following ig administration of CQ to CS and CR infected mice was determined by HPLC method (extra standard). RESULTS: 3 h after 4.06 mg/kg and 400 mg/kg administration, the accumulation of CQ in both infected RBC was similar, but in CR was 54.0% and 42.1% less than in CS (P < 0.001) respectively. The CQ accumulation in CS 3 h and 7 h after ig administration of CQ 400 mg/kg was similar (0.780 +/- 0.128 nmol/protein and 0.760 +/- 0.180 nmol/mg protein), while in CR the CQ-accumulation was higher at 7 h than that at 3 h after CQ administration (0.452 +/- 0.079 nmol/mg protein and 0.559 +/- 0.124 nmol/mg protein P < 0.02). CONCLUSION: The low CQ accumulation in CR is attributable to the parasite rather than to the infected RBC, and CR was not found to excrete CQ more rapidly than CS. PMID- 12078241 TI - [The morbidity investigation of residents in a highly endemic village of schistosomiasis in Poyang Lake region]. AB - AIM: To investigate the morbidity of residents in a highly endemic village of schistosomiasis, and to evaluate the usefulness of ultrasound in morbidity study. METHODS: Morbidity induced by Schistosoma japonicum was evaluated and compared in target population by using parasitological examination, ultrasound investigation and medical history questionnaire. RESULTS: The schistosome infection rate was 22.9% in the study population and the prevalence of abnormal ultrasound findings in the liver and spleen was high. The abnormal ultrasound findings correlated with sex significantly, the abnormality of liver parenchyma correlated with age, sex, infection status, occupation and the number of treatment after multiple logistic regression analysis. CONCLUSION: The ultrasound examination could find out the direct pathological changes resulting from schistosome infection. PMID- 12078242 TI - [Application of improved western blot method in diagnosis of cysticercosis]. AB - AIM: To evaluate the diagnostic value of four specific antigens from cDNA of Cysticercus cellulosae. cC1, cC2, and cP1 and cH1 (28 kDa, 18 kDa, 14 kDa and 34 kDa), mixing in equal proportions for the diagnosis of cysticercosis. METHODS: Taking the FP (fusion proteins) as antigen to make IWB (improved Western blot) analysis basing on the detection of antibody responses against FP, and making ELISA/IHA-crude antigen(CA) analysis. They were evaluated comparatively while using 107 infected sera of cysticercosis cases, 40 infected sera of clonorchiasis cases, 24 infected sera of echinococcosis cases and 34 sera of healthy persons. The FP are encoded by cDNAs of beta-galactosidase-specific antigens of Cysticercus cellulosae isolated from the cDNA library. RESULTS: 94(87.9%) sera from 107 cysticercosis cases recognized FP in IWB and could not cross-react with the sera of echinococcosis cases, clonorchiasis cases and healthy persons, the specific rates were 100%, whereas ELISA, IHA using CA were 84.1% and 74.8%, respectively and could cross-react with the sera of echinococcosis cases, the false positive rates were 2.5% and 12.5% respectively; CA-ELISA/IHA could cross react with the sera of clonorchiasis patients, the false positive rates were 8.3% and 16.7%, respectively; and they could also cross-react with the sera of healthy persons, the false positive rates were 8.8% and 11.8%, respectively. CONCLUSION: The recombinant FP used in the immunodiagnosis of cysticercosis is specific and sensitive. PMID- 12078243 TI - [Characteristics of development and morphology of Echinococcus granulosus of camel origin in north Xinjiang]. AB - AIM: To study the characteristics of development and morphology of E. granulosus of camel origin in north Xinjiang. METHODS: Six dogs were infected with protoscolices and then purged with arecoline at days 35 and 45 p.i. RESULTS: Six dogs were heavily infected, the worm burden were 14,780 to 135,900 at days 35 and 45 p.i. At day 35 p.i. 77% of the had developed three segments, 10% contained mature eggs with fully developed "thickshelled" embryophores. Morphological studies revealed the following characteristics for 35-day-old worms; the mean length of the terminal segment accounted for 56% of the total worm length; the position of the sexually mature segment was always terminal (77%); the female reproductive system possessed an enlongated ovary with compact lobules; the Mehlis gland was covered by the vitelline gland and the testes were distributed throughout the segment arranging in a row posterior to the vitelline gland. CONCLUSION: The E. granulosus of camel isolate in north Xinjiang can readily be distinguished from the horse, sheep, cattle and pig strains, but is similar to the camel (Camelus dromedarius) strain distributed in North Africa. PMID- 12078244 TI - Ultrastructural observation on spermatocytogenesis in Taeniid cestodes. AB - AIM: To study the spermatocytogenesis of taeniid cestodes at the ultrastructural leaves. METHODS: Transmission electron microscopy. RESULTS: The ultrastructural observation on spermatocytogenesis in Taenia solium, T. saginata and T. pisiformis were made by TEM. Two types of spermatogonia; type A and B, as well as the supporting cells surrounding the peripheral of spermatogonia are recognized. The type A spermatogonia are stem cells and the type B are mother cells which produce 16 primary spermatocytes by mitosis for 4 times with the cells unseparated. The primary spermatocytes are characterized by the ribosome masses in the cytoplasm. 32 secondary spermatocytes arranged in roselike were produced by reductive division of primary spermatocytes. The secondary spermatocytes become the spermatid quickly by short time development. CONCLUSION: The dividing mode of spermatogonia in Taeniid cestodes is mitosis with cells unseparated. PMID- 12078245 TI - [Effect of RNA against hepatic fibrosis in rabbits infected with Schistosoma japonicum]. AB - AIM: To study the effect of ribonucleic acid (RNA) against schistosomal hepatic fibrosis in rabbits. METHODS: 54 rabbits were randomly and equally divided into normal group, infected group, and RNA group. On the 40, 70, 100th day after RNA i.m. injection, 6 rabbits from each of the above 3 groups were killed and their plasma and liver were examined by way of biochemistry and pathology. RESULTS: Compared with the infected group the collagen fiber of liver in RNA group was less produced, the hepatic cellular nucleolus was enlarged, the number of rough endoplasmic reticulum (RER) and the granules on the RER were increased, the collagen fiber around fat-storing cell was decreased, the content of hyaluronic acid in plasma, the hydroxyproline level in liver tissue, the percentage of collagen fiber distribution and the collagen fiber level in liver tissue of RNA group were significantly reduced. CONCLUSION: RNA might play a role in the prevention of schistosomal hepatic fibrosis. PMID- 12078246 TI - [Establishment of a method for detecting transforming growth factor beta 1 mRNA]. AB - AIM: To establish a method to measure the TGF-beta 1 mRNA level for studying the mechanism of fibrogenesis caused by schistosomiasis japonica. METHODS: Reverse transcription polymerase chain reaction and dot blot analysis were used. A plasmid of TGF-beta 1 was constructed for standardization, and beta-actin was used as control. Eight different concentrations of the plasmid and 11 double-tube of TGF-beta 1 mRNA in the peripheral blood mononuclear cell (PBMC) of different persons were measured. RESULTS: Quantitative results of 8 different dilutions of TGF-beta 1 plasmid had positive with the logarithm of the original concentration. The results of the 11 double-tube were 1.71 +/- 0.90 and 1.54 +/- 0.88. CONCLUSION: The duplicability and stability of the method showed it can be used to analyse the TGF-beta 1 mRNA level of the peripheral blood mononuclear cells. PMID- 12078247 TI - [Studies on DNA vaccines of parasites]. PMID- 12078248 TI - [Application of isoenzyme pattern of allelic gene of isoenzymes in the taxonomic study of trematodes and their snail hosts]. PMID- 12078249 TI - [Current status of studies on biosystematic and malaria transmission of Anopheles dirus in China]. PMID- 12078250 TI - [DNA vaccine of Plasmodium falciparum: immune response of BALB/c mice induced by injection of the recombinant plasmid DNA]. AB - AIM: To observe the level of immune responses of BALB/c mice induced by direct injection of the recombinant plasmid DNA and to provide evidence for the application of DNA vaccine of Plasmodium falciparum (P. f.) in animals and humans. METHODS: The recombinant plasmid pcDNA3-Pf8 encoding the hybrid antigens of PfCMR-MSP1(19) was constructed. The presence of Pf8 plasmid was detected in the muscle, liver, kidney, heart, spleen and lung by PCR. ELISA, T lymphocyte transformation test, inhibitory assay of the growth and development of P. f. in vitro were used to determine the level of humoral and Th-cell responses. RESULTS: pcDNA3-Pf8 was amplified by PCR technique in the different tissues from BALB/c mice which were injected with pcDNA3-Pf8. The immune serum was specifically recognized by the soluble antigens of P. falciparum (titer: 1:2,560 by ELISA). The specific proliferative response of spleen T cells from immunized mice was remarkably higher than that of control group, the transformation rate being up to 32.82 +/- 3.64%. The immune serum could also inhibit the growth and development of the erythrocytic stages of P. falciparum in vitro. CONCLUSION: Direct injection with the recombinant plasmid pcDNA3-Pf8 specifically elicits BALB/c mice to generate humoral and cell-mediated immune responses, the resulting immune serum being inhibitory to the growth and development of P. f. in vitro. PMID- 12078251 TI - [Detection of Cryptosporidium parvum in fecal samples using polymerase chain reaction combined with non-radioactive labeled DNA probe hybridization]. AB - AIM: To develop a non-radioactive labeled probe hybridization technique for the detection of the PCR products of Cryptosporidium parvum DNA in fecal samples and evaluate its specificity and sensitivity. METHODS: DNA of C. parvum was prepared directly from C. parvum infected patients' fecal samples by the direct lysis method, and was used as the template for PCR. A pair of oligonucleotide primers was synthesized and used to prime the amplification of a 452 bp target fragment specific for C. parvum. The PCR products were directly spotted or transferred on to NC membrane. Then, the PCR products were hybridized with the biotin-labeled oligonucleotide probe and coloured by a substrate BCIP (5-bromo-4-chloro-3 indolyl-phosphate). RESULTS: Positive hybridized signals were only showed in the PCR products of DNA extracted from the fecal samples of C. parvum infected patients, but not in the other DNA of P. yoelii, L. donovani, G. lamblia, C. albicans, E. coli and human leucocytes. The lowest detectable amount of both dot or Southern blotting hybridization was 0.1 pg of C. parvum DNA. CONCLUSION: A combination of PCR and non-radioactive biotin-labeled probe hybridization assay is highly sensitive, specific, and relatively simple and safe for the detection of the PCR products of C. parvum in fecal samples. PMID- 12078252 TI - Effect of testosterone on Leishmania donovani infection levels of murine bone marrow derived-macrophages. AB - AIM: To investigate the effect of the male sex hormone, testosterone (Te), on Leishmania donovani infection levels of bone marrow derived macrophages(BMMs) from female mice of strain C57BL/6J. METHODS: After three weeks of Te-treatment, the BMMs were isolated, challenged with L. donovani at a ratio of 10 to 1 promastigotes per macrophage, and the infection levels of different time points were monitored by Giemsa staining. RESULTS: BMMs from Te-treated mice had a significantly increased initial uptake(3 h post infection, P < 0.05) of promastigotes and carried heavier infection levels at all time points(24 h, 48 h, 72 h post infection, P < 0.01), compared with those from oil treated controls. CONCLUSION: Te can increase L. donovani infection levels of BMMs, being possibly related to Te-induced immunosuppression. PMID- 12078253 TI - [Sequencing and homology analysis of cDNAs encoding FhGST and AsGST]. AB - AIM: To study the homologies between FhGST, AsGST and Sj26GST. METHODS: Total RNAs were extracted from Fasciola hepatica (Fh) and Ascaris suum (As) by guanidine hydrochloride/CsCl ultra-centrifugation. Two pairs of primers were designed according to FhGST and AsGST sequences. RT-PCR was carried out using total RNA as template. PCR products were directly sequenced. Then the homology analysis was made using DNASIS software on nucleotide level. RESULTS: A FhGST403 bp fragment and a AsGST411 bp fragment were obtained and they both located in the open reading frame. CONCLUSION: On nucleotide level, the homologies are 59% and 54% between the two cloned fragments and Schistosoma japonicum 26 kDa GST (Sj26GST) respectively. PMID- 12078254 TI - [Studies in immunological diagnosis of Pneumocystis carinii infection]. AB - AIM: To assess the diagnostic value of Pneumocystis carinii(PC) antibodies and circulating antigens detection in alveolar lavage fluid (ALF) and serum. METHODS: The immunosuppressive rat model was used in this study. The PC antigens in both ALF and sera were detected by double sandwich ELISA, and the specific IgG antibody levels in sera were detected by IFA at different times after immunosuppression treatment. The detection results were compared with the corresponding results of PC finding in lung impression smear. RESULTS: The PC antigens in ALF were positive in all infected rats 6-8 weeks after immunosuppression while in control rats were all negative. By comparison, the PC antigens detected from sera were negative in most infected rats. During immunosuppression treatment with steroid, antibody titers in the infected rats rose slightly in some rats. After steroid was withdrawn, PC antigens in ALF became negative gradually but antibody titers rose progressively with time in all rats. CONCLUSION: Detecting PC antigens in ALF may be useful for diagnosing PC infection but these antigens were difficult to be detected in serum. The rise in serum IgG antibody does not necessarily indicate an existing infection. PMID- 12078255 TI - [Ultrastructure of Schistosoma japonicum sperm]. AB - AIM: To understand the ultrastructure of Schistosoma japonicum sperm. METHODS: Testis and oviduct were cut from male and female of S. japonicum respectively, location was assured by observing with microscope. Specimens preparation for transmission electron microscope were made by traditional way. RESULTS: The sperm of S. japonicum includes two parts: a head and a single tail. The elongated ovoid head measures by 6.2 x 1.4 microns, lacking an acrosome, rounding anteriorly and tapering posteriorly. A prominent anterior mass of mitochondria lies beneath the plasma membrane. A layer of microtubes under the plasma membrane are parallel with long axis of the head. A coil of longitudinal microtubes were present beneath the plasma membrane. The nucleus is largely electron-opaque with some electron-lucent patches or less electrondense material. The tail is a single flagellum with a unique axoneme, which originated from a centriole. The axoneme in main part of the flagellum (including middle and posterior parts of the tail) consists of nine doublets and a single diffuse central structure, forming 9 x 2 + <<1>> type. But the axoneme in the transition area (anterior part of the tail) lacks central material, forming 9 x 2 + 0 type. CONCLUSION: The sperm ultrastructure in the schistosomes is homogeneous, but differs distinctly from those trematodes of other Digenea. PMID- 12078256 TI - Ultrastructural observation on the transformation of the spermatozoon in spermatogenesis of Taeniid cestodes. AB - AIM: To study the transformation of taeniid spermatozoon during spermatogenesis on the ultrastructure level. METHODS: Transmission electronmicroscopy. RESULTS: This paper was the second part of the ultrastructural observations on the spermatogenesis in Taeniid cestodes: The transformation of the spermatozoon. The 64-spermatid-plasmodium was generated from the secondary meiosis of the 32 rose like secondary spermatocytes. The transformation of the spermatozoon was a complex process. Firstly, both the cytoplasm and nucleus of the spermatid elongated while the cytoplasm increased and connected to the cytoplasmic mass (cytophore). Secondly, the chromatins of nucleus polymerized to a thread-bundle like structure. Finally, it detached from the cytoplasm mass and became a mature spermatozoon. The mature spermatozoon was a thread-like structure, about 16.2 18.6 microns in length and 0.35-0.45 micron in diameter. There were two structurally distinct parts: the part with the nucleus was the head and the part without the nucleus was the tail. The head measured about 5-6 microns in length, 1/3 of the total body length and contained an elongated nucleus which twined around the axoneme without mitochondria. The tail was about 11.2-16.6 microns in length. Throughout the core of the tail was an axoneme with typical "9 + 1" structure. The anterior of the tail, just behind the head, were some mitochondria, about 1.6-1.7 microns in total length which twined around the axoneme. The posterior of the tail contained only an axoneme. In cross-section of the spermatozoon, about 46 microtubules were beneath the plasma membrane. CONCLUSION: The transformation of the spermatozoon is a very complex process. PMID- 12078257 TI - [Detection of Brugia malayi larva in mosquito vector by PCR and PCR-ELISA]. AB - AIM: To develop sensitive, specific, simple assays for the detection of Brugia malayi larva in mosquito vectors. METHODS: The optimum conditions for PCR and PCR ELISA were determined. With dissected larvae, detection limits and as well as specificities of the two assays were worked out. The optimized assays were then tested with infected and non-infected mosquito vectors, Anopheles sinensis. RESULTS: Both PCR and PCR-ELISA detected DNA equivalent to a single L1-stage larva(L1). While the actual detecting limits of the two, as estimated and titrated respectively, reached to 1/10 and 1/100 of a L1. When the two assays were used to detect the larvae isolated from 113 infected A. sinensis, all gave specific bands and positive reactions. However, trials on direct amplifications with crude extracts of infected mosquito vectors consistently failed to give specific bands upon electrophoresis and negative results in PCR-ELISA as well. CONCLUSION: Both PCR and PCR-ELISA were preliminarily established for the detection of isolated B. malayi larva in mosquito vector, which proved to be sensitive, specific and easy to manipulate. PMID- 12078258 TI - [Preparation and characterization of monoclonal antibodies against Cryptosporidium parvum]. AB - AIM: To develop and characterize the monoclonal antibodies(McAbs) against Cryptosporidium parvum (C. parvum). METHODS: BALB/c mice were immunized with purified oocyst antigens of C. parvum from human isolates and McAbs against C. parvum were prepared by using hybridoma technique. The characterization of McAbs was studied by means of indirect immunofluorescent assay (IFA) and Western blot analysis. RESULTS: Four clones of hybridoma cell lines secreting McAbs against oocyst antigens of C. parvum were obtained. The immunoglobulin subclasses of these four McAbs were identified as IgM (Z4C8,Z3D7) and IgG1(Z2B6,Z3D2) and their light chains were kappa chain. IFA results showed that three McAb (Z4C8, Z3D7 and Z3D2) conjugates produced a diffuse fluorescence in the Cryptosporidium sporozoites (CSP) and one (Z2B6) produced local fluorescence in the oocyst wall of C. parvum. Four McAbs could recognize 4 of 42 polypeptide antigen bands of CSP with molecular weights of 20.5 kDa, 33 kDa, 60.5 kDa and 95 kDa, among which both Z4C8 and Z3D7 could recognize 20.5 kDa band that was defined as a principal antigenic protein of CSP. CONCLUSION: These four McAbs possess their own characteristics of recognizing antigenic epitopes on CSP, but react specifically against antigens of C. parvum. PMID- 12078259 TI - [Experimental studies on dynamic changes of circulating antigens of Schistosoma japonicum in infected rabbits]. AB - AIM: To observe the dynamic changes and disappearance of three kinds of schistosome circulating antigens(CAgs) in infected rabbits before and after praziquantel treatment. METHODS: Dot-ELISA assays were developed by using 3 McAbs to determine the dynamic changes of the gut-associated antigen (GAA), membrane associated antigen (MAA) and soluble egg antigen (SEA) respectively in sera of rabbits infected with S. japonicum. RESULTS: By the 4th week after infection 39% of the rabbits infected with S. japonicum were GAA positive and only 8.6% were MAA or SEA positive. All the infected rabbits becOme positive with GAA, MAA and SEA by the 8th week after infection. The titers of the GAA, MAA and SEA in sera of the infected rabbits reached their peaks by the 10th week after infection. Titers from rabbits of the untreated group rabbits remained high level until the 27th week after infection. SEA titers were found negative in 10% of the infected rabbits in the treatment group by the 6th week after treatment. The titers of GAA of, MAA' and SEA were found negative in 20%, 80% and 70% of the rabbits in the treatment group, respectively the 8th after treatment. GAA, MAA and SEA in sera except one of the treated rabbits were all negative by the 14th week after treatment. CONCLUSION: The determination of GAA is useful for the early diagnosis of schistosomiasis, while the determination of, MAA and SEA might be used for the efficacy evaluation. PMID- 12078260 TI - [The role of village doctors and their influence factors in malaria control]. AB - AIM: To consolidate the acquired achievements of malaria control and to work out an effective programme for training the village doctors in Yibin District. METHODS: A stratified sampling was done at random with sampling 27 village medical stations and 67 village doctors from 9 townships of 3 counties. A questionnaire concerning malaria control knowledge of the village doctors and the quality of their carrying out antimalaria measures was used in this study. RESULTS: The accuracy rate of answering the questionnaire among the village doctors was 79.3%, being higher in the village doctors receiving antimalaria training than in those not receiving antimalaria training. 82.5% of the doctors could correctly complete the tasks of controlling malaria. In addition, the quality of malaria control implementing in areas where the doctors and their stations had timely got investment funds, preferential policy, system of reward and subsidy from local governments, was superior to those areas where the doctors and their stations had not received those on time. 91.9% of the trained doctors completed the tasks of controlling malaria better than untrained ones. CONCLUSION: Greater efforts should be made to give a systematic training to village doctors on the knowledge and technique of malaria control, and to strengthen management and support in order to help these doctors accomplishing the tasks of malaria control better. PMID- 12078261 TI - [SurveilLance on filariasis after its basic elimination in Hunan Province]. AB - AIM: To explore the fluctuation pattern of prevalence and to detect the residual infection in Hunan Province where filariasis had been basically eliminated since 1986. METHODS: Longitudinal surveillance and cross-sectional surveillance were extensively carried out by using parasitological, entomological and serological methods in areas previously endemic for filariasis. RESULTS: In 55 counties, cross-sectional surveillance on filariasis had been executed for 11-19 years. The results showed that microfilaria cases had not been found for 4 years, and filaria larvae had failed to be detected from mosquito vectors for 12 years. Serological surveillance in the population revealed that the mean positive rate of IFAT had dropped from 13.15% in 1987 to 1.06% in 1996, the latter rate being similar to that of the nonendemic area. 6 longitudinal surveillance spots in 5 counties (cities) had been observed for 10-17 years. Annual quantitative blood examination of 11 microfilariemia cases showed that 10 cases became negative in 1 10 years, and the remaining 1 case remained positive at the 14th year. A. sinensis were dissected with no filaria larvae found in 2 surveillance spots of malayan filariasis. Culex quinquefasciatus were dissected with filaria larvae found only in the first to third year, the natural infection rates were 0.38% 1.98% in 4 surveillance spots of bancroftian filariasis. CONCLUSION: Since the basic elimination of filariasis in Hunan Province in 1986, the number of residual microfilariemia cases decreased year after year, suggesting that the transmission of filariasis has been interrupted. PMID- 12078262 TI - [Electrophoretic analysis of soluble antigens of a pathogenic isolate of Trichomonas vaginalis]. AB - AIM: To analyze the parasite soluble antigens of a pathogenic isolate of Trichomonas vaginalis. METHODS: The soluble antigens of a pathogenic isolate of T. vaginalis isolated from a patient suffered from trichomonad vaginitis was studied by means of SDS-PAGE followed by densitometric analysis and immunoblotting. RESULTS: A total of 14 distinct protein bands was demonstrated using a 10% resolution gel concentration. Densitometrogram indicated that the soluble antigens with MWs of equal to and below 72 kDa accounted for 89.5% of the total volume with the dominant fractions of 48-51 kDa which accounted for 19.2%. Having MWs over 100 kDa accounted only for 7.4%, while the lowest M. W. fraction demonstrated was found to be at the site of 13 kDa. By immunoblotting, the specific anti-T. vaginalis antibodies raised in mice recognized a broad spectrum of molecular size range of these antigenic fraction. However, no immunological reaction was seen at the 86 kDa site. CONCLUSION: The proteins of MWs 48-51 kDa possess a larger protein content and a higher immunological reactivity. PMID- 12078263 TI - [Ingestion of marine fish and anisakiasis]. PMID- 12078264 TI - [Advances in the study of diagnostic antigens in clonorchiasis]. PMID- 12078265 TI - [Studies on malaria transmission among blood donors]. PMID- 12078266 TI - [Current status of diagnostic reagents for schistosomiasis in China--an analysis based on results from parallel examinations and check-up of methods/reagents]. PMID- 12078267 TI - [The role of L-arginine and L-citrulline in activated macrophage against Toxoplasma gondii infection in vitro]. AB - AIM: To determine whether both the production of NO and the toxoplasmastatic or toxoplasmacidal activity of activated macrophages cultured in physiologic levels of arginine can be enhanced by increasing the availability of arginine, or citrulline. METHODS: After the activated M phi were infected with the RH strain of T. gondii, the levels of NO production, the infection rate of M phi and the number of tachyzoites in parasitophorous vacuoles were determined after the M phi were cultured in medium added with various concentrations of L-arginine or L citrulline for 18 h. RESULTS: (1) The multiplication of intracellular toxoplasmic tachyzoites could be inhibited by activated macrophages, depending on the production of NO in the presence of physiologic levels of arginine. (2) Increased exogenous arginine or citrulline resulted in a significant elevation of NO production induced by activated macrophages and further reduction of infection rate of macrophages as well as inhibition of multiplication of intracellular tachyzoites. (3) Citrulline could fully substitute for arginine in enhancing NO production and toxoplasmastatic or toxopasmacidal activity. CONCLUSION: The physiological levels of arginine were able to induce sufficient intrinsic NO production to inhibit intracellular multiplication of toxoplasmatic tachyzoite but unable to protect cell from infection and that increasing the substrate levels for NO biosynthesis may enhance in vitro toxoplasmastatic or toxoplasmacidal activity of activated macrophages. PMID- 12078268 TI - [Genotyping of Plasmodium falciparum isolates by amplification of glutamate-rich protein gene using polymerase chain reaction]. AB - AIM: To develop a genotyping method based on amplifying glutamate-rich protein (GLURP) gene for the diagnosis and identification of Plasmodium falciparum. METHODS: Two pairs of primers specific for GLURP gene of P. falciparum were designed and synthesized. R2 polymorphic domain of GLURP gene was amplified by nested PCR, which was applied to genotyping of P. falciparum isolates obtained from patients attending the malaria clinic at the village of Borai, Thailand. RESULTS: Conspicuous polymorphism of GLURP alleles in natural populations of P. falciparum was found. 290 GLURP alleles were detected in 154 P. falciparum infections. Among the above-mentioned alleles, 12 different GLURP genotypes were distinguished according to different DNA sizes. Of them, the most frequently found allele was a variant of 770 bp, the least allele was that of 1,100 bp. More than 43% of the patients were found to be infected with mixed alleles. No apparent change for frequencies of the 12 different alleles was found in the 9 month longitudinal study. CONCLUSION: A genotyping method is developed for the research of strain taxonomy and pathogenesis of malaria parasites. PMID- 12078269 TI - In vitro and in vivo effect of levopraziquantel, dextropraziquantel versus racemic praziquantel on different developmental stages of Schistosoma japonicum. AB - AIM: To compare the antischistosomal effect of racemic praziquantel (Pra) and its enantiomers, levopraziquantel (L-Pra) and dextropraziquantel (D-Pra), on different developmental stages of Schistosoma japonicum. METHODS: The in vitro effects of the drugs were determined in different stages of schistosomes maintained in RPMI 1640 supplemented with 20% calf serum. In vivo study mice infected with schistosome cercariae were treated intragastrically (ig) with Pra, L-Pra or D-Pra at different intervals after infection. The efficacy of the drugs was evaluated by residual mean worm number. RESULTS: Based on the degree of tegument damage induced by L-Pra, d28 and d35 schistosomes were most susceptible to L-Pra, while d14 schistosomules being least susceptible. At comparable concentrations of 0.1-1 g/ml, L-Pra was more active than Pra even when the concentration of L-Pra was reduced to one-half of the minimum effective concentration of Pra. At above-mentioned concentrations D-Pra exhibited no apparent in vitro effect on different stages of schistosomes. When infected mice were treated ig with L-Pra, Pra or D-Pra at a single dose of 300 mg/kg or 500 mg/kg, only the former two drugs showed apparent effect on d0, d21, d28 and d35 schistosomes and less or much less effect on d3, d7 and d14 schistosomules. D-Pra only exhibited a negligible effect on d35 adult schistosomes as compared with L Pra and Pra. When mice infected with d35 adult schistosmes were treated ig with L Pra 150 mg/kg, the efficacy was similar to that of mice treated with Pra 300 mg/kg. CONCLUSION: L-Pra is the principal active component against schistosomes in racemic Pra. PMID- 12078270 TI - [Cloning and sequence analysis of the gene encoding the partial region CS protein of a Plasmodium falciparum isolate from Yunnan]. AB - AIM: Determining nucleotide sequence of the circumsporozoite protein partial gene of the Plasmodium falciparum PFD-3/YN (Yunnan of China) and finding out the differences of the CS gene sequence between Chinese Plasmodium falciparum isolate and other isolates. METHODS: The circumsporozoite protein gene fragment was amplified by polymerase chain reaction and cloned into M13 bacteriophage. M13-CSP single strand DNAs of the three positive clones were extracted respectively. Then, the nucleotide sequence of the CS gene fragment was determined by the dideoxy chain termination method. PCGENE software was used to compare and analyze the CS gene sequence of the six isolates. RESULTS: Different degrees of diversity of the CS gene sequences were found among P. falciparum PFD-3/YN and other isolates(T4, Wellcome, NF54, 3D7 and 7G8). A non-silent substitution at the nucleotide level being found in the P. f Th/Tc antigenic epitope region. CONCLUSION: There were differences in the CS gene sequence among P. falciparum PFD-3/YN and those of other isolates. PMID- 12078271 TI - [Studies on the life cycle of Paragonimus heterotremus]. AB - AIM: In order to clarify the biological features of the lung fluke, Paragonimus heterotremus, it is necessary to replicate its complete life cycle in the laboratory. METHODS: Tricula wumingensis infected with the miracidiae of P. heterotremus were regularly dissected after infection for larvae observation. The cercariae of P. heterotremus from positive snails were used for experimental infection of the crab, Samanniathelphusa nanningensis. The crabs were then examined for the metacercariae at regular intervals. The infection of mammals (cats and rats) was performed with the metacercariae obtained from experimentally infected crabs. RESULTS: The miracidia were hatched by incubating the eggs in water (room temperature 20 degrees C to 31 degrees C) for 16 days to 21 days. The epidermal cells of the miracidia were arranged in 4 transverse rows, 6 in the first row, 7 in the second, 3 in the third, and a single terminal cell at the posterior end of the body. The cells of the first row have indentation in their bases. Mature sporocysts, first generation rediae and second generation rediae(containing dissociative cercariae), were found on the 26th day, 41st day and 58th day after infection of the snails respectively(room temperature 21 degrees C-30 degrees C). The sporocyst had a birth pore at its anterior tip. The length ratio of the gut to the body was 1:3 in the first and 1:6 in the second generation rediae. The mature second generation rediae contains 5-21 mature cercariae which emerge from its pharynx. The cercaria has general characteristics of Paragonimus. Its flame cell pattern was formulated as 2[(2 + 2 + 2) + (2 + 2 + 2)] = 24. After the infection of crabs with cercariae, the mature metacercariae were found in the crabs on the 60th day. The crabs could be successfully infected by either: giving mature cercariae orally to them or raising them in water with positive snails. After the metacercariae obtained from experimental crabs were given to cats and rats, P. heterotremus adult worms were recovered from the worm cyst in their lungs. The morphology of various larval stages and adult worms were described. CONCLUSION: The life cycle of P. heterotremus was completed for the first time in the laboratory. The whole life cycle from egg to adult worm has been described in the present study. PMID- 12078272 TI - [Establishment of germinal cell line of Echinococcus granulosus]. AB - AIM: To establish a cell line of Echinococcus granulosus. METHODS: The proliferating membranes striped from liver cysts in a naturally infected sheep from Urumq were released to monodispersed germinal cells by grinding. The germinal cells were cultivated in the RPMI 1640 medium supplemented with 10%-20% calf serum on collagen-coated 24-well culture plate alternately from passage 1 to passage 14, and then continued. The morphological feature and growth situation were observed by light microscopy. The cultured cells were inoculated into BALB/c mice to identify the infectivity. ELISA was used to determine the immunogenicity of the cells. RESULTS: The germinal cells have been cultivated continuously up to passage 75. The subcultured cells were circular in shape with smooth surface and had the tendency to form syncytia and tissue-like masses. The cells from this cell line could be stored for at least 15 days in refrigerator at 4 degrees C and 10 months in liquid nitrogen. No cyst materials were detected in the mice inoculated with cells. The antigens from cell line could react with positive sera from mice infected with protoscoleces and sera against secreted antigens of cyst membrane, soluble antigens from cyst membrane, soluble antigens from protoscoleces and SHF. CONCLUSION: A germinal cell line of Echinococcus granulosus was successfully established. PMID- 12078274 TI - [Immunodiagnosis of cysticercosis by simplified western blotting using recombinant fusion protein as antigen]. AB - AIM: To provide simple and useful method for the immunodiagnosis of cysticercosis. METHODS: Fusion protein of beta-galactosidase-Cysticercus cellulosae cDNA was analysed with simplified Western blot. RESULTS: The positive rate was 93.5% when detecting 124 cases of human cysticercosis sera with four clones (cC1, cC2, cP1 and cH1) and 100% when detecting 38 cases of porcine cysticercosis sera with three clones (cC1, cC2 and cP1), being higher than those using monoclone fusion protein alone. Moreover, the fusion proteins did not react with other parasitosis sera. CONCLUSION: Simplified Western blot analysis using recombinant fusion protein as antigen was highly sensitive and specific and easy to be operated. PMID- 12078273 TI - [Studies on dynamic changes of SOD and MDA in cultured cells from Schistosoma japonicum schistosomula]. AB - AIM: To observe the dynamic changes of superoxide dismutase(SOD) and maldondialdehyde(MDA) in cultured cells from Schistosoma japonicum schistosomula and to explore the degeneration process of cultured cells. METHODS: Pyrogallol method and thiobarbituric acid method were used to determinate the levels of SOD and MDA, respectively, in the cells cultured for 0-54 days. RESULTS: SOD in the cells reached a peak level on d12 of the early stage of culture and then decreased. The MDA levels in the cultures cells ascended gradually, and had two peaks on d18 and d48, during the period of culture. CONCLUSION: The cultured cells from S. japonicum schistosomula degenerated progressively. PMID- 12078275 TI - Ultrastructural observations on the formation and metabolism of calcareous corpuscles in Cysticercus cellulosae. AB - AIM: To study the formation and metabolism of calcareous corpuscles from Cysticercus cellulosae at the ultrastructure level. METHODS: Transmission electron microscopy. RESULTS: The developmental processes of calcareous corpuscles could be divided into two stages: the intracellular formation stage and the extracellular metabolic stage. The calcareous corpuscles were formed in a cell which we named calcareous corpuscle forming cell. At the early stage of the formation, the corpuscles appeared to be secretory granules in the cells. With the development of the corpuscles, they became bead-shaped and lamellae-like, then the calcareous corpuscle forming cell enlarged and the organellae degenerated. Finally the corpuscles gathered to form particle substances with black dense background, while the nucleus and organellae of the forming cell all disappeared. There were 1-3 or 10-20 calcareous corpuscles in a mature forming cell. Then, the corpuscles were released to the parenchymal tissues and gradually appeared to be concentric lamella or an empty cavity during the metabolic process. CONCLUSION: The calcareous corpuscles were formed in calcareous corpuscle forming cell and consumed in metabolic process in the parenchymal layer of Cysticercus cellulosae. PMID- 12078276 TI - [Observation on the infectivity of different densities of Plasmodium vivax to Anopheles sinensis]. AB - AIM: To study the infectivity of different densities of P. vivax to An. sinensis. METHODS: Venous blood samples from 38 volunteers artificially infected with P. vivax of South-Yunnan collected at the time of initial parasitaemia, primary attack and relapse, were used to infect Anopheles sinensis by feeding on parasitaemia blood in vitro. The finding of oocysts in the mid-gut of the mosquito by microscope was taken as criteria of positive infection. RESULTS: P. vivax in the initial parasitaemia did not infect the mosquito. In the cases of primary attack, when the density of parasite was more than 100/microliter blood, the infectivity and intensity rose with the prolongation of the course of the disease and the increase in the density of the parasite. The infectivity of the P. vivax in the relapse cases was higher than in the primary attack cases, the mosquitoes could be infected at a parasite density of 1/microliter blood. The mosquitoes acquired a higher infection rate, a higher positive rate of mosquito mid-gut and a higher oocyst index in the primary attack cases whose parasite density were higher than 1,000/microliter blood and in the relapse cases whose parasite density were higher than 100/microliter blood. CONCLUSION: The P. vivax in the period of clinical attack was one of the most dangerous period of the spread of malaria. The cases of relapse were the more dangerous infective sources in malaria transmission. PMID- 12078277 TI - [Screening and characterization of Schistosoma japonicum reinfection-related cDNA clones]. AB - AIM: To obtain cDNA clones coding for S. japonicum (Sj) reinfection-related specific immunologic molecules for vaccination or diagnosis. METHODS: An adult S. japonicum cDNA library, of which the cloning efficiency is 3.13 x 10(6), with directional inserts in the vector lambda gt11, was screened with pooled serum from 43 reinfected human cases in a schistosomiasis-endemic area with high level of Sj specific-lgG4 antibodies. Primers located several bases away from the insert position on the lambda gt11 arm were used to amplify the insert fragments by standard PCR. SILVER SEQUENCE (PCR-silver) DNA sequencing system was then used to sequence selected positive clones with relatively large inserts and homology comparison with GenBank database was also carried out. RESULTS AND CONCLUSION: Eleven phage plaques were defined as positive out of the 8.6 x 10(4) phage plaques immunoscreened by human specific lgG4 antibody. GenBank database retrieval showed that 4 new Sj gene clones including Sj reinfection-related specific protein-encoding gene and Sj mitochondrial protein-coding gene have been obtained. PMID- 12078278 TI - [Continuous cultivation of a large number of Plasmodium falciparum gametocytes in carbon dioxide incubator]. AB - AIM: To establish a method for continuously cultivating a large number of P. falciparum gametocytes in vitro using carbon dioxide incubator. METHODS: The number of gametocytes produced in experimental and control groups was compared after the addition of various concentrations of NaHCO3 to the culture medium. RESULTS: The gametocytes began to rise on day 5 of cultivation and reached a peak on day 11-13. The peak gametocyte loads were 1.9% +/- 0.6% and 1.3% +/- 0.4% (P < 0.05) in experimental and control group, respectively, indicating that the complete medium with 30 mmol/L sodium bicarbonate was beneficial to gametogenesis. The numbers of stages I-V gametocytes rose to a peak on d5, d7, d11, d13 and d15, respectively. On day 15 the percentage of stage V gametocytes was 7.1%-52.6% with an average of 24.3%. The ratio of macrogametocyte to microgametocyte was 12.8:1. The parasites were able to produce high gametocytaemia up to 24th subculture after thawing. Laboratory reared Anopheles stephensi fed through membrane on blood infected with P. falciparum were dissected, no oocyst was found in the midgut. CONCLUSION: A culture system which could consistently and stably produce a large number of gametocytes of P. falciparum was established. PMID- 12078279 TI - [Detection of Toxoplasma gondii DNA in human lymph node tissue by in situ hybridization]. AB - AIM: To detect the presence of Toxoplasma gondii in lymph node tissue in patients with Toxoplasma infection. METHODS: T. gondii (RH strain) specific DNA fragment clones were obtained by using PCR and gene recombination technique. The DNA fragments used as hybridization probes were labelled with digoxigenin by random primer method. The technique of in situ hybridization (ISH) was used to detect T. g DNA in the lymph node sections. RESULTS: Four out of 120 samples T. g DNA were found positive, one with Hodgkin's disease (HD) (1/32), one with non-Hodgkin's lymphoma (NHL) (1/41) and 2 with chronic lymphadenitis (CL) (2/47). The total positive rate was 3.3%. It was demonstrated that this highly specific probe could detect 10 pg of the total RH strain T. g DNA. CONCLUSION: ISH was applicable in detecting pathogens in the lymph node tissues of individuals with Toxoplasma infection. PMID- 12078280 TI - [An epidemiological survey of schistosome cercarial dermatitis among the residents living along the banks of the Huaihe River system]. AB - AIM: To make an epidemiological survey of schistosome cercarial dermatitis among the residents living along the banks of the Huaihe River. METHODS: Making inquiries about signs and symptoms of schistosome cercarial dermatitis, and physical examinations of manifestation in the residents (fishermen and peasants). Cercariae shed from the Radix auricularia collected in the field were used to infect ducklings. The eggs and miracidia separated from the feces of the full grown ducks bred by fishermen were used to infect Radix auricularia. RESULTS: Schistosome dermatitis locally known as "duck itch" (meaning duck fecesdermatitis) was discovered among the residents living along the Huaihe River bank. The main signs and symptoms of the dermatitis included diffuse erythema or red papular eruption with areorae, urticae, urticant itch or titillation around the affected areas (including thoracic part, abdomen and lower limbs etc). The cercariae with eye spot were found from Radix auricularia collected from Huaihe riverside and were identified to be the species of Trichobilharzia can both hatch miracidium, which laboratory-reared Radix auricularia were infected with miracidia hatched from the two types of eggs, rhomboid eggs and crescent eggs, found in the feces of full grown ducks bred by fishermen and shed the same cereariae with eye spot. Besides, adult worms of Trichobiharzia were obtained by dissecting both experimentally infected ducklings bred in the laboratory and naturally infected ducks bred in the Huaihe river-side area. The above-mentioned eggs, miracidia, cercariae and adult worms were all identified to belong to the life-phase of Trichobilharzia. CONCLUSION: The epidemic disease known as schistosoma cercarial dermatitis harmful to the health of the residents living along the banks of Huaihe River is caused by the cercariae of Trichobilharzia. PMID- 12078281 TI - [Survey of human parasite species in China (continued)]. PMID- 12078282 TI - [Immunization of mice with native tropomyosins from Schistosoma japonicum and Oncomelania hupensis]. AB - AIM: To study the protection against cercariae challenge in mice immunized with native tropomyosin (TM) antigens from Schistosoma japonicum (Chinese strain) and Oncomelania hupensis. METHODS: The native TM antigens were extracted from adult worms of S. japonicum (SjcTM) and head-pad of O. hupensis (OhTM), the purity and the molecular weight of these antigens were analysed by SDS-PAGE. Each C57BL/6 female mouse was immunized subcutaneously by injection of 5 micrograms SjcTM or 30 micrograms OhTM emulsified in an equal volume of Freuds adjuvant for three times. Control mice were injected only with adjuvant. All the mice were artificially challenged with cercariae of S. japonicum at five days after final injection. Six weeks after challenge, the mice were perfused, the worms and the eggs collected from the livers and the intestines of mice were counted. RESULTS: The prepared SjcTM and OhTM antigens were shown to be pure, the molecular weight being around 40 kDa. Compared with the control group, the worm reduction rates of SjcTM group and OhTM group were 21.1% and 28.2%, respectively; the egg reduction rates per worm pair in the tissues were 20.1% and 52.2% (liver), 32.8% and 28.1% (intestine), respectively; the matured egg reduction rates in the tissues were 25.9% and 46.8% (liver), 39.7% and 66.8%, respectively. CONCLUSION: SjcTM and OhTM antigens can be served as new candidates of vaccine against schistosomiasis japonica. PMID- 12078283 TI - [Cloning and sequencing of the gene coding the sexual stage antigen Pfs48/45 of Plasmodium falciparum]. AB - AIM: To express the antigen Pfs48/45 in vitro and provide an antigen for the development of the transmission-blocking vaccine. METHODS: According to the published nucleotide sequence of Pfs48/45 of Plasmodium falciparum isolate NF54, a pair of oligonucleotides was designed and used as primers(P1, P2). The gene encoding the gametocyte/gamete-specific membrane protein Pfs48/45 of P. falciparum isolate FCC1/HN has been amplified by using polymerase chain reaction (PCR) technique. The PCR product was purified and directly sequenced by the dideoxynucleotide terminator method with 5'-end primer P1. At the same time, the purified PCR product was digested with BamHI and EcoRI and cloned into the plasmid pcDNA3, then the recombinant clones were transformed into E. coli strain TG1. The recombinant plasmid pcDNA3-Pfs48/45 was screened and identified by PCR amplification and restriction analysis. RESULTS: 1. The gene fragment Pfs48/45 was specifically amplified from the genomic DNA of Plasmodium falciparum isolate FCC1/HN; 2. The sequence demonstrated that the that the 5'-end nucleotide and predicted aminoacid sequence of Pfs48/45 from FCC1/HN isolate was basically identical with that from NF54 isolate. We found that the sequence of the Pfs48/45 gene from the isolate FCC1/HN differs from the published sequence(isolate NF54) only at positions 307 and 372(T-->C). The substitution of T-->C at the position of 372 generates a new restriction site Taq I. The PCR product digested by Taq I generates DNA fragment of 984 bp and 379 bp, suggesting that the PCR product is the gene encoding the transmission blocking antigen Pfs48/45; 3. The gene fragment of Pfs48/45 was directly inserted into the BamHI and EcoRI site of plasmid pcDNA3. CONCLUSION: The nucleotide sequence of Pfs48/45 of Plasmodium falciparum isolate FCC1/HN from south China was similar to that of isolate NF54. The recombinant plasmid pcDNA3-Pfs48/45 was successfully constructed, providing a means to evaluate the role and biological function of this sexual-stage-specific protein of Pfs48/45. PMID- 12078284 TI - [Cross-protection against Schistosoma japonicum infection in mice immunized with Trichinella spiralis muscle larva antigen]. AB - AIM: To explore the cross-protection against S. japonicum cercaria challenge in mice immunized with T. spiralis larva antigen. METHODS: Groups of BALB/c mice were immunized with 4 preparations of T. spiralis larva antigen, respectively. Vaccinated mice were then challenged with 30 and 100 S. japonicum cercariae, respectively. Forty-five days later, all mice were sacrificed and examined for adult worm burden, liver and stool egg count. RESULTS: All groups of mice immunized with 4 preparations of T. spiralis larvae antigen without adjuvant showed significant protection against S. japonicum. Among these antigen preparations, the soluble antigen (TsLSA) presented highest protection with reductions in worm burden and liver and stool egg counts of 21.3%, 48.0% and 58.5%, respectively. When mice were immunized with TsLSA plus Freunds complete adjuvant (FCA) or with double-dose of the antigen, the worm reduction rate was increased to 29.3% and 39.6%, respectively. CONCLUSION: Different preparations of T. spiralis muscle larva antigen could induce protection against S. japonicum in mice. PMID- 12078285 TI - [Cloning and expression of Schistosoma japonicum eggshell protein gene]. AB - AIM: To construct an eukaryotic expression system pcDNA3/ESG-HeLa cell and explore the possibility of whether the eggshell protein might be used as potential targets for vaccine development. METHODS: The eggshell protein gene (ESG) was amplified using PCR technique and inserted into vector pcDNA3 to construct recombinant vector pcDNA3/ESG. The expression of the eggshell protein was performed in HeLa cell line. SDS-PAGE, dot-ELISA and Western blotting were used to identify the antigenicity of the expressed product. RESULTS: The amplification of ESG (618) bp was achieved and high level expression of the eggshell protein was obtained in HeLa cells and the amount of total expressed products was up to 24.4%. Dot-ELISA and Western blot analysis demonstrated that the expressed protein could be recognized by sera from rabbits infected with S. japonicum and from female worm- or egg-immunized rabbits. In addition, the expressed protein could specifically stimulate proliferative response of the immunized BALB/c mouse spleen cells, the transformation rate being up to 44.57%. CONCLUSION: The eukaryotic expression system pcDNA3/ESG was successfully established and high level expression of the eggshell protein of S. japonicum was achieved in HeLa cell line. The expressed recombinant protein exhibited immunological activities. PMID- 12078286 TI - [Effect of dihydroquinghaosu on the development of Plasmodium yoelii yoelii in Anopheles stephensi]. AB - AIM: To observe the effect of dihydro qinghaosu (DQHS) on the development of Plasmodium yoelii yoelii in Anopheles stephensi and to explore the possibility of whether DQHS has preventive effect against malaria. METHODS: Plasmodium yoelii yoelii-infected mice that had been administered with a single dose of different dosages (60 mg/kg, 120 mg/kg, 180 mg/kg and 240 mg/kg) of DQHS were exposed to mosquitoes to suck blood. The development of malaria parasites in the mosquitoes of the control group and the treated group were observed by light microscopy (LM) and transmission electron microscopy (TEM). RESULTS: DQHS exhibited certain inhibitory effect on the infectivity of gametocyte. The degree of inhibition was related to the developmental stages of gametocyte and the drug dosage. Immature gametocytes were more sensitive to DQHS than the mature gametocytes. With the increases in dosage, the positive rate and the density of oocyst and sporozoite decreased. However, the difference in the density of sporozoite between 180 mg/kg and 240 mg/kg treated groups was insignificant. TEM result showed damage of the membrane system and vacuolation in the cytoplasm of the oocysts (12-13 d) on mosquito midgut of 60 mg/kg 16 h treated group. The oocysts in the mosquitoes continued to develop after treated with 120 mg/kg of DQHS for 16 h on 3-day instar. There was no significant difference (P > 0.05) in the density of oocyst and sporozoite between the control group and the treated group. CONCLUSION: DQHS can affect the infectivity of gametocyte and decrease the malaria transmission but cannot inhibit the sorogonic stage directly. PMID- 12078287 TI - Karyotype analysis of Leishmania isolates in China. PMID- 12078288 TI - [Analysis of nuclear DNA gene types of Leishmania isolates from hilly and plain foci of China]. AB - AIM: To analyse the nuclear DNA (nDNA) polymorphism of Leishmania isolates from hilly and plain foci of China. METHODS: nDNA were analysed by endonuclease digestion, Southern blotting and chromosomal localization. Probes were labeled with digoxigenin. RESULTS: Using gp63 gene probe, similar hybridization bands were found to be existed between nDNAs of Leishmania donovani Jiangsu human isolate and L. d. Jeddah, also between nDNAs of L. d. Sichuan human isolate and L. infantum. Using beta-tubulin gene probe, there were two similar hybridization bands existed between nDNAs of L. d. Jiangsu human isolate and L. d. Jeddah, and three similar hybridization bands existed between nDNAs of L. d. Sichuan canine isolate and L. d. Gansu canine isolate, and two similar hybridization bands existed between nDNA of L. d. Sichuan canine isolate and L. d. Wenchuan human isolate. CONCLUSION: Homology exists between L. d. Jiangsu human isolate and L. d. Jeddah from plain foci, between L. d. Sichuan human isolate from hilly foci and L. infantum, between L. d. Sichuan canine isolate and L. d. Gansu canine isolate from hilly foci. Homology as well as differences exist between L. d. Sichuan canine isolate and L. d. Wenchuan human isolate. Heterogeneity exists between isolates from hilly foci and plain foci. PMID- 12078289 TI - [Correlation between the dose and the anti-Toxoplasma effect of activated mouse macrophages induced by IFN-gamma and the synergism between IFN-gamma and TNF alpha]. AB - AIM: To find out the dose dependence of the anti-Toxoplasma effect induced by IFN gamma and to determine the possible synergistic activity between TNF-alpha and IFN-gamma. METHODS: The in vitro effect of cultivated mouse peritoneal macrophages activated by IFN-gamma alone or IFN-gamma combined with different doses of TNF-alpha on the intracellular tachyzoites of RH strains and the nitric oxide (NO) level in the culture medium supernatant were simultaneously determined. RESULTS: With the increase in the dose of IFN-gamma, the anti Toxoplasma effect was augmented and the NO level was enhanced. At 24 hours after tachyzoite invasion, a significant reversed correlation was demonstrated between the NO level and the number of intracellular parasites. CONCLUSION: The anti Toxoplasma effect of macrophages activated by IFN-gamma appears to be dose dependent and TNF-alpha acts synergitically with IFN-gamma in the activation of macrophages. The production of reactive NO could be an important effector in the IFN-gamma primed anti-Toxoplasma action. PMID- 12078290 TI - [Detection of HFRSV in Eulaelaps shanghaiensis and Ornithonyssus bacoti by using in situ hybridization]. AB - AIM: To provide molecular biological evidence of transmission of hemorrhogic fever with renal syndrome virus (HFRSV) by gamasid mites, Eulaelaps shanghaiensis and Ornithonyssus bacoti. METHODS: Frozen sections of the gamasid mites 10 days and more than 100 days after biting suckling mice inoculated with HFRSV were in situ hybridized with dig-labelled HFRSV cDNA probes. RESULTS: RNA was detected in frozen sections of Eulaelaps shanghaiensis, after biting suckling mice inoculated with Hantaan (76-118) and Seoul (UR) virus, respectively. Most of the fine granules of the virus RNA were located in the nuclei, cytoplasm and nuclear membrane of cells of brain cortex, caeca and genitalia of the mites. In situ hybridization results showed that 17 of 31 mites in Hantaan group and 10 of 23 mites in Seoul group were positive. The virus RNA was still detected in tissues of the mites on d132 for Hantaan group and on d102 for Seoul group after infection, respectively. Among 20 Ornithonyssus bacoti detected 12 were positive on d10 after biting suckling mice inoculated with Hantaan virus, and the virus RNA was mainly found in the cells of genitalia (Figs. 1-13). CONCLUSION: Both Eulaelaps shanghaiensis and Ornithonyssus bacoti could be infected with HFRSV by biting HFRSV-positive mice. E. shanghaiensis could be infected with both Hantaan and Seoul virus, and the two types of HFRSV were found to be maintained in the mites for 132 days and 102 days, respectively. These confirmed that E. shanghaiensis and O. bacoti are suitable vectors and reservoirs of both Hantaan and Seoul virus and might play an important role in the cross transmission of the two types of HFRSV. PMID- 12078291 TI - [Studies on the experimental transmission of Rattus-borne Hantavirus by Ornithonyssus bacoti]. AB - AIM: To demonstrate the role of rat mite (Ornithonyssus bacoti) in the transmission of Rattus-borne hemorrhagic fever with renal syndrome (HFRS). METHODS: In the transmission experiments, about 100 O. bacoti per pool were isolated and placed in a jar, unfed for 4 d at 23 +/- 1 degrees C. Suckling Wistar rats inoculated with Hantavirus strain Z45 or Seoul virus strain UR were placed in each jar for free attack by the mites for 12 hours. After 14 d the normal suckling Wistar rats were bitten by the mites. Fifteen days later, the lung tissues and sera of the infected rats were collected and detected for Hantaviral antigen by indirect fluorescent antibody technique (IFAT). For demonstration of the infection of O. bacti with Rattus-borne Hantavirus PCR technique was applied to detect Rattus-borne Hantaviral RNA. RESULTS: Sukling Wistar rats inoculted Hantavirus strain Z45 or Seoul virus strain UR were bitten by O. bacoti and then these mites were fed on 4 and 5 of normal suckling rats in each jar, respectively. The antigens of Hantavirus strain Z45 were positive in all the lungs of the normal rats bitten by the mites, the sera titers of the rats were from 1:10 to 1:40. The antigens were positive in 3 of the 4 rats, the sera titers were from 1:20 to 1:40. Both of the viruses could be maintained in O. bacoti for 22 days. The blocking test showed when 1:30 Hantavirus immunosera were exposed to the lung samples and then reacted with the sera from the patients with HFRS, all the specific fluorescence reactions of the samples were blocked, whereas the control group including the normal rat lung tissues and sera were all negative (Fig. 1). CONCLUSION: O. bacoti might play a role as the vector of HFRS and a reservoir host as well. PMID- 12078292 TI - [Evaluation of the effect of causes of death of advanced schistosomiasis patients on the life span of population by using potential years of life lost analysis]. AB - AIM: To evaluate the effect of the causes of death of advanced schistosomiasis patients on the life span of population in an area where schistosomiasis had been under control by applying the indicators of potential years of life lost (PYLL) and rate of PYLL (PYLLR). METHODS: Using the formula of calculation of PYLL and PYLLR, the causes of death of 487 patients of advanced schistosomiasis in 3 previously endemic townships of Shanghai suburb. RESULTS: Of 487 cases who died between 1955 and 1995, the PYLLR of all causes of death was 256.3@1000. The main causes of death were hepatic failure (100.3@1000), liver carcinoma (43.4@1000) and cancer of other organs (24.8@1000). In the 1960s, the PYLLR of advanced schistosomiasis complicated with hepatic failure and upper digestive tract hemorrhage was 222.2@1000 and 34.5@1000, the PYLLR of the complications of schistosomal liver cirrhosis, hepatic failure and hemorrhage accounted for 67.5% of the total PYLLR, being apparently higher in the non-splenectomy group than in the splenctomty group. After the schistosomiasis was under control, both PYLL and PYLLR decreased linearly. As compared with the 1960s, the PYLLR of all causes of death and the causes of death of the complications of schistosomal liver cirrhosis in the 1990s has declined by 62.9% and 83.2%, respectively, the PYLLR of upper digestive tract hemorrhage was found only 0.3@1000 in the last 5 years. CONCLUSION: PYLL and PYLLR analysis could be used to quantify the effect of complications of advanced schistosomiasis and other causes of death on the life span of population. PMID- 12078293 TI - [Application of dot immunogold filtration assay for diagnosis of Schistosomiasis japonica]. AB - AIM: To quest for a simple and convenient assay for the diagnosis of schistosomiasis. METHODS: Three monoclonal antibodies against different epitopes of antigens of S. japonicum were used to develop a dot immunogold filtration assay for detection of schistosomal antigen-antibody reaction on the membrane during the filtration process. The results could be read with the naked eye within a few minutes. RESULTS: The sensitivity of the SEA assay was 16 ng/ml. In 69 serum samples from patients with chronic schistosomiasis, the positive rate and specificity of this assay were 60.8% and 95.2%, respectively. Dot-ELISA was used in detecting 137 serum samples from patients with chronic schistosomiasis, the positive rate and specificity were 54.7% and 94.6%, respectively. When sandwich-ELISA was used in detecting 118 serum samples from patients with chronic schistosomiasis, the positive rate and specificity were 61.9% and 95.7%, respectively. CONCLUSION: The sensitivity and specificity of DIFA were similar to those of ELISA, but DIFA required less time and no special equipment. PMID- 12078294 TI - [Exploration on statistical method for calculating intensity of the infection in residents in nationwide sampling survey of schistosomiasis]. AB - AIM: To set up statistical methods for calculation of intensity of schistosome infection in residents in nationwide sampling survey of schistosomiasis. METHODS: Based on the epidemiological survey of schistosomiasis and according to the medical statistics for multiple stratifying and clustering random sampling, the calculation method on intensity of schistosome infection was inferred. RESULTS AND CONCLUSION: Nine formulas were worked out for the calculation of the intensity of schistosome infection in different endemic provinces with different ecotypes. These formulas are of practical value. PMID- 12078295 TI - [In vitro sensitivity of Plasmodium falciparum to chloroquine, piperaquine, pyronaridine and artesunate in Yuxi prefecture of Yunnan province]. AB - AIM: To assess the sensitivity of Plasmodium falciparum to four antimalarials. METHODS: WHO standard in vitro microtest was used. RESULTS: The resistance rates of the malaria parasite to chloroquine, piperaquine pyronaridine and artesunate were 85.7%, 66.7%, 38.1% and 5.0%, respectively. CONCLUSION: Among the antimalarials tested in Yuxi Prefecture, Yunnan Province, high resistance of P. falciparum to chloroquine and piperaquine was found. The sensitivity of P. falciparum in part of the cases to pyronaridine decreased. However, most of the cases were relatively sensitive to artesunate. PMID- 12078296 TI - [Current status of research on chloroquine-resistant falciparum malaria]. PMID- 12078297 TI - [Discussion on two ecological aspects of Paragonimus westermani cercaria]. PMID- 12078298 TI - [Study on early repair of destructive hand injury and reconstruction of hand function]. AB - OBJECTIVE: To investigate a clinical method in repairing destructive hand injury and reconstructing hand function in early stage. METHODS: From January 1990 to June 1999, composite tissue transplantation was used to repair destructive hand damage and reconstruct hand function with vascular anastomosis in emergency (33 cases) or subemergency (126 cases). For the radial damage, combined flaps with toes was used to treat cicatricial contracture of thumb-web space. Opposing function of thumb was reconstructed by combined transplantation of short extensor muscle of great toe, short extensor muscle of toes or short abductor muscle of great toe in the same time of freeing the second toe. For no or deficient anastomosing vessels in donor site, "Y"--shaped reversal vein, anterograde bridging, or branches bridging of host vessels were applied. RESULTS: All of tissue transplantation were survived, including combined tissue transplantation in 51 cases, composite tissue transplantation in 4 cases, tissue transplantation after replantation of severed wrist and finger in 4 cases, there were no infection or necrosis in all cases, and hand function recovered well. CONCLUSION: It is an ideal procedure to repair destructive hand injury and reconstruct hand function by combined or composite tissue transplantation with vascular anastomosis in emergency or subemergency. PMID- 12078300 TI - [Importance of the posterior and lateral arterial network of elbow on the super regional and mutual-pedicled axial flap]. AB - OBJECTIVE: To explore the importance of the posterior and lateral arterial network of elbow in the application of the super-regional and mutual-pedicled axial flap. METHODS: Twenty-seven upper extremities of adult cadavers were prepared as casts of Acrylomintril Batradiene Styrene(ABS) resin and corroded in a strong solution of NaOH according to natural layers of human tissue. The source, site and structure of the posterior and lateral arterial network of elbow were observed, the number and total sectional area of anastomosing branches crossing the line between two humeral epicondyles were measured and compared with the medial and anterior region. RESULTS: There are 8.64 +/- 2.74(36.42%) and 8.30 +/- 1.19(35.0%) anastomosing branches crossing the posterior and lateral regions, and total section areas are (0.48 +/- 0.11) mm2 and (0.37 +/- 0.03) mm2 respectively. So there is very rich arterial network around the elbow. CONCLUSION: The enough number of anastomosing branches and their section areas of the posterior and lateral region of the elbow make it possible to connect super regional and mutual-pedicled axial flaps crossing the elbow. PMID- 12078299 TI - [Experience of application of skin flap in repairing 112 patients with severe thermo pressure injury of hand]. AB - OBJECTIVE: To investigate the clinical effect of skin flaps repairing severe thermopressure injury of hand. METHODS: From January 1989 to December 1998, 112 patients with severe thermopressure injury of hand were repaired by various skin flaps transfer, the size of skin flaps was 6 cm x 8 cm to 12 cm x 18 cm. Postoperative patients were treated by combined rehabilitation in early stage. RESULTS: All the flaps were survived with satisfactory effect. Sixty-six patients were followed up 6 to 12 months, skin flaps all showed better colour and texture, and function of the hand was satisfactory. CONCLUSION: Different skin flaps are adopted to repair severe thermopressure injury of the hand according to different skin defects of the hand, combined early rehabilitation treatment, to achieve good recovery of function and appearance of the hand to the greatest extent. PMID- 12078301 TI - [Transplantation of free scapular flap to repairing injury of faciocervical region]. AB - OBJECTIVE: To sum up transplantation of free scapular flap to repairing injury of faciocervical region, 103 cases in past 16 years were reviewed. METHODS: The scapular flap contained the ascending, descending, and transverse branches of the circumflex scapular artery. The range of the flap was as following: across the scapular spine to the acromion in upper edge, to the level of anterior iliac spine in lower edge, to the bilateral middle axillary line in both sides. RESULTS: The clinical result was good. Fifty eight cases were followed up for 3 months to 10 years, the function of the faciocervical region was recovered with satisfied contour. In part of the patients, a second-stage operation was performed to make the flap thinner. CONCLUSION: The maximal range of the flap can be 45 cm x 16 cm. For its sufficient blood supply, it should be of no influence on survival of the flap if the window is open in suitable site. Because of its large range of donating region and sufficient blood supply, it is fit for children. PMID- 12078302 TI - [Repair of severe radiation-induced ulcer]. AB - OBJECTIVE: To explore a method to repair severe radiation-induced ulcer and evaluate its clinical effects. METHODS: From August 1988 to December 1998, 12 cases of severe radiation-induced ulcer were treated by muscular or musculocutaneous flap transfer, which included 1 case of pectoralis major muscular flap, 3 cases of three-lobed transverse rectus abdominis musculocutaneous flap, 2 cases of latissimus dorsi musculocutaneous flap, 1 case of tensor fascia lata musculocutaneous flap, 1 case of longitudinal rectus abdominis musculocutaneous and tensor fascia lata musculocutaneous flap, 1 case of gastrocnemius muscular flap, 1 case of gluteus major musculocutaneous flap, huge filleted musculocutaneous flap from lower limbs in 1 case and from upper limbs in 1 cases. RESULTS: All the flaps survived. Primary healing occurred in 9 cases and secondary healing in 3 cases. The extensive multiple ulcers in 2 cases were repaired with filleted limb musculocutaneous flap, and patients were saved. And in other 3 cases, the chest wall defect as the result of radionecrosis were repaired with simultaneous breast reconstruction. CONCLUSION: Muscular or musculocutaneous flaps have abundant blood supply, they are ideal tissue to fill the cavity and repair the defect, especially in repairing severe radiation induced ulcers. PMID- 12078303 TI - [Modified breast reconstruction by latissimus dorsi musculocutaneous flap]. AB - OBJECTIVE: To investigate the effect of breast reconstruction with latissimus dorsi musculocutaneous flap. METHODS: Since 1994, 60 cases were performed breast reconstruction with latissimus dorsi musculocutaneous flap with fat tissue nourished by thoracodorsal artery according to the shape and volume of the normal breast on the other side. All of cases were followed up for 3 months to 5 years. RESULTS: Among the 60 cases, excellent effect was obtained in 41 cases (68.3%), good effect in 16 cases (26.7%), unsatisfactory in 3 cases (5.0%). CONCLUSION: Modified latissimus dorsi musculocutaneous flap to reconstruct breast overcome the shortcoming of volume deficiency of traditional latissimus dorsi in breast reconstruction, and it is a safe and easy-manipulated surgical operation. PMID- 12078304 TI - [Color-flow duplex Doppler scanning study in the tram flap perforators: a report of 94 consecutive patients]. AB - OBJECTIVE: To investigate the location of transverse rectus abdominis musculocutaneous (TRAM) flap perforators on abdomen skin and the peak systolic flow velocities of the perforators. METHODS: A series of 94 consecutive patient's TRAM flap perforators were detected by color-flow duplex Doppler scanning peroperatively. Perforator locations were tabulated on the abdomen skin according to their vertical position relative to the umbilicus and their lateral location relative to the abdominal midline. RESULTS: The perforators on both left and right side of TRAM flap were quite not symmetry. The peri-umbilical region contained 81.1% of the perforators. All perforators were detected with peak systolic flow velocities ranging from 5 to 81 cm/s. CONCLUSION: The preoperative color-flow Doppler localization of TRAM flap perforators improves the surgeon's ability to design the flap in order to capture the dominant vessels, and select single- or double-pedicle and free TRAM flaps based on each patient's individual characteristics. PMID- 12078305 TI - [Anatomical study and clinical applications of flaps based on cutaneous branches arising from posterior tibial vessels]. AB - OBJECTIVE: To provide an anatomical basis for the clinical applications of the medial fascinocutaneous flap of calf and to verify its clinical value. METHODS: In 20 lower limb specimens of adult human cadavers, the number, course, size, position and distribution of septocutaneous branches of the posterior tibial vessels are marked by means of red latex and black ink irrigations through femur artery and septocutaneous branches of the posterior tibial artery respectively. RESULTS: The posterior tibial artery gives off several septocutaneous branches at the upper, middle and lower one-third of the leg respectively. Each septocutaneous artery has one or sometimes two concomitant veins. Based on this result, anterograde or reverse pedicled fasciocutaneous flap can be performed for the purpose of repairing soft tissue defects of leg and foot. The flap was clinically applied to treat leg and foot soft tissue defects in 12 cases with satisfactory results. CONCLUSION: The flap is easy to dissect, the posterior tibial artery can be preserved with high successful rate. Therefore, it offers an useful alternative in the repairing and reconstruction of nonextensive soft tissue defects in the leg and foot. PMID- 12078306 TI - [Clinical application of distal base sural island flap]. AB - OBJECTIVE: To sum up the experience of clinical application of distal base sural island flap. METHODS: From January 1997 to April 1999, the posterior island flap of leg pedicled with sural nerve and its nutrient vessels was applied to repair heel or dorsum of foot in 6 cases, chronic ulcer of heel in 2 cases, chronic osteomyelitis in 2 cases, scar contracture of bone defect accompanying fistulation in 1 case. The range of flap was 5 cm x 8 cm to 8 cm x 12 cm. RESULTS: All the flaps survived completely without vascular crisis. All the wounds healed by first intention. Followed up 3 to 12 months, no ulcer, osteomyelitis, fistulation were recurrence, and the sensation of flap was recovered slightly. CONCLUSION: The flap do not damage critical blood vessels and nerves, the donating region is hidden. The manual of operation is simple and blood supply of flap is sufficient. It can repair the defect of soft tissue on heel and dorsum with high survival rate. PMID- 12078307 TI - [Clinical application of sural nerve island flap pedicled with collateral vessels]. AB - OBJECTIVE: To sum up the application experience of the sural nerve island flap pedicled with the collateral vessels. METHODS: From 1997, the retrograde-flow sural nerve island flaps pedicled with collateral vessels were performed to repair the soft tissues defects of the shank in 3 cases, ankle in 3 cases and foot in 8 cases. RESULTS: Twelve flaps were survived, one flap was partially necrosed and one flap was necrosed. Among them, 10 wounds healed by first intention, 3 cases were healed after changing dressing and the one necrosed flap was repaired by free flap transplantation. Nine cases were followed up for 3 to 21 months and had fine appearance and function. The flap texture was similar to normal skin, the sensation of flap partially recovered after 6 months. CONCLUSION: The flap has more reliable blood supply and great rotation arc, it is easy to resect with little injury. It is excellent for repairing the soft tissues defect in the anterior leg, ankle and proximal half of foot. It is more significant while the main blood vessels are damaged. PMID- 12078308 TI - [Extraction and purification of Schwann cells cytoplasmic neurotrophic proteins with high pressure liquid chromatography and study on its neurobiological activities]. AB - OBJECTIVE: To purify and study Schwann cells cytoplasmic neurotrophic protein. METHODS: The dissociated SC taken from 300 newborn rats sciatic nerves were cultured, collected, ultrasonicated and ultraspeed centrifuged. The supernates were ultrafiltrated and concentrated by using ultrafiltration units with PM10, PM30, PM50 ultrafiltration membranes. The ultrafiltrated-concentrated solution with the protein molecular weight 10-30 ku, 30-50 ku and > 50 ku were collected respectively. The dissociated spinal cord motoneurons of 14 days embryonic rats were cultured with serum-free conditional medium and the additional SC cytoplasmic proteins were added into the medium. The results showed that the 10 30 ku and > 50 ku SC cytoplasmic proteins were able to maintain the survival of motoneurons for 24 hours. Then the 26 ku and 58 ku proteins were further extracted and purified from SC cytoplasm by high pressure liquid chromatography, and their neurobiological activities were studied. RESULTS: The 26 ku and 58 ku Schwann cell's cytoplasmic proteins were able to maintain the survival of motoneurons cultured in the serum-free medium for 48 hours. The highest biological activity concentration is 20 ng per well. CONCLUSION: Schwann cells cytoplasm contains motoneuron neurotrophic proteins with molecular weight 26 ku and 58 ku. PMID- 12078309 TI - [Primary surgical operation in the treatment of varicosis of lower limb accompanied by chronic leg ulcer]. AB - OBJECTIVE: Retrospective clinical analysis of primary surgical operation in the treatment of lower limb accompanied by chronic leg ulcer were adopted in this study. METHODS: From September 1990 to June 1998, there were 31 males and 20 females, aged 68 years in average, the area of ulcer varied from 5 cm x 3 cm to 22 cm x 11 cm. The ligation and strip of saphenous vein, debridement and free skin flap grafting were finished in primary operation. RESULTS: The skin flaps were survived completely in 50 cases, only 1 case was necrosis partially and healed after changing dressing. Forty-two cases were followed up for 16 months to 9 years (66 months in average), the varicosis and ulcer were healed in 39 cases and only 3 relapsed in ulceration. CONCLUSION: Primary surgical operation in the treatment of varicosis of lower limb accompanied by chronic ulcer is practicable in clinic. The curative efficacy is satisfactory and the operative manipulation is simple. PMID- 12078310 TI - [Surgical intervention in the treatment of skin defect of hell]. AB - OBJECTIVE: To investigate the efficacy of different flaps in the treatment of skin defect of hell. METHODS: Forty-six patients with skin defect of hell were adopted in this study. There were 39 males and 7 females, 29 years old in average. Six different flaps were applied in the reconstructive operation, 14 plantaris medialis flaps, 4 flexor digitorum brevis muscle flaps, 3 abductor hallucis flaps, 7 latissimus dorsi flaps, 16 distal medialis flaps of leg pedicled with the cutaneous branch of posterior fibial artery, 2 foot dorsum flaps. RESULTS: All the flaps survived, primary healing of the wound in 45 cases and secondary healing in 1 case. Followed up for 3 months to 4 years, 43 patients obtained good flap sensation, the function of weight bearing were satisfied in 43 patients. CONCLUSION: The six different flaps should be applied according to patient's condition individually. The sensation of flap is very important to the function of weight bearing. PMID- 12078311 TI - [Expression of type I collagen and its receptor system in osteoblasts]. AB - OBJECTIVE: To study the expression of type I collagen and its receptor system integrin alpha 2 beta 1 in different passages of osteoblasts. METHODS: The expression of type I collagen and integrin alpha 2 beta 1 in the primary, sixth and fifteenth passage of osteoblasts were detected by S-P immunohistological staining technique, and their mRNA expression by quantity RT-PCR technique. RESULTS: Type I collagen and integrin alpha 2 beta 1 were expressed in different passages of osteoblasts and there was no significant difference among three passages by immunohistological technique. Their mRNA expression was gradually decreased with subculture. CONCLUSION: Type I collagen promotes the adhesion and phenotype expression of osteoblasts through its receptor-integrin alpha 2 beta 1. The reductive expression of type I collagen-receptor system will decline the phenotype of osteoblasts. PMID- 12078312 TI - [Short tandem repeat loci examination after repair of coracoclavicular ligament injury by tissue engineered tendon]. AB - OBJECTIVE: To evaluate the result of clinical application in the repair of coracoclavicular ligament injury by tissue engineered tendon using the technique of short tandem repeat loci examination. METHODS: In september 1999, human embryonic tendon cells and artificial materials were co-cultured in vitro to construct tissue engineered tendon, which repaired coracoclavicular ligament injury. After 6 months of operation, micro-tissue were sampled during the operation of removal of internal fixation, and morphological characteristics were examined by HE staining, DNA of tissues were extracted to examine D3S1754 and Cyar04 gene loci. RESULTS: The shoulder function of the patient was recovered well after operation, and no local or systemic immunological rejection were occurred. The electrophoresis typing showed 13/14 at D3S1754 and 8/9 at Cyar04 in the tissue of tissue engineered tendon, while the autogenous ligament were 13/13 and 8/8 at D3S1754 and Cyar04 loci respectively, which suggested that the tissue engineered tendon was survived in vivo. CONCLUSION: The examination of short tandem repeat loci is a better index to evaluate the survival situation of tissue engineered tissue after transplantation in clinical application. PMID- 12078313 TI - [Tissue engineering of skin]. AB - OBJECTIVE: To review the recent progresses on tissue engineering of skin. METHODS: Recent original articles about tissue engineering of skin were extensively reviewed, which focused on the progresses and major problems concerning the epidermal substitutes, dermal substitutes, cultured-epidermal composite skin graft. RESULTS: Most investigators had come to conclusion that the optimal skin substitute should provide for immediate reconstruction of both the lost epidermis and dermis. The research was mainly focused on how to transplant epidermal cells immediately, preserve their activity and function, and develop the extracellular matrix which could effectively accelerate the function of transplanted cells, induce vascular growth from the wound bed, could be biodegradable, no toxicity and no danger of carrying pathogen. CONCLUSION: The major research trends of tissue engineering of skin should be focused on the study of immediate transplantation of epidermal cells, accelerate wound healing and developing extracellular matrix of dermis. PMID- 12078314 TI - [Clinical evaluation of frozen-phalanx-joint-tendon-sheath composite tissue allograft]. AB - OBJECTIVE: To evaluate clinical result of reconstructed thumb and finger with a free hallux nail flap(HNF) and frozen-phalanx-joint-tendon-sheath composite tissue allograft in 270 cases. METHODS: The patients were followed up with reexamination in the ambulant clinic, communication, X-ray photography, lab examination, isotope 99mTc MDP and reoperation. The data were analyzed by statistics or proved by clinical observation, which were followed up for five years in average (ranging from five months to sixteen years). RESULTS: Enveloping the allogeneic finger composite tissue with self-HNF and pieces of phalanx of great toe, it could reconstruct a thumb or finger with good contour and nutrition. The excellent rate of opposition function of the reconstructed thumbs was 71.91%. The sense of the fingers recovered after 3 months to 8 months of operation. Two-point discrimination was 3 mm to 15 mm. The junction between implanted allo-phalanges and auto-phalanges could be hastened by implanted with vascularized autogenous phalanx pieces in the HNF. The isotope 99mTc MDP was used to take X-ray photography in 24 cases for four months to 9 years and seven months, which showed that the blood vessels grew into the allo-phalanges. However, the Charcot's arthropathy of allogeneic joints and bony absorption still could be seen in some cases. That might be concerned with chronic abrasion of joint or chronic rejection of host to graft. CONCLUSION: The operation is fit for repairing the defect of thumb or finger in any degree. The implanted vascularized self-phalanx pieces can promote bone union, but it can not prevent the allogeneic joints from arthropathy or bone absorption. PMID- 12078315 TI - [Development of prefabricated flaps]. AB - OBJECTIVE: To sum up the experimental development and clinical application of prefabricated flap. METHODS: The reported experimental results and clinical application of prefabricated flap extensively reviewed. RESULTS: Previous studies had proved that the revascularization of prefabricated flap mainly through anastomoses of implanted vessels and the original vessels of the flap, the implanted vessels slowly formed a new and complete blood vessel network, which could dominate the whole flap, three to four weeks later, the new vessels were mature and the flap could be transferred. Clinically, the superficial temporal vessels, gastroepiploic vessels, circumflex femoral vessels and thoracodorsalis vessels could be harvested for prefabricated flap with satisfactory results. CONCLUSION: Prefabricated flap provides a new method for the treatment of complicated defects. PMID- 12078316 TI - Lifelong learning. PMID- 12078317 TI - Annual report details health care fraud and abuse enforcement. PMID- 12078318 TI - Developmental considerations for resuscitation of the VLBW infant. AB - The survival rates for very low birth weight (VLBW) infants continue to increase while the morbidity rates show little improvement. Negative outcomes have been associated with hypoxia, hyperoxia, hypocarbia, barotrauma, intraventricular hemoRrhages, metabolic acidosis, cold stress, and overwhelming stimuli. Protection from iatrogenic harm must always be provided for the extremely fragile infant. Standards for resuscitation of the VLBW infant in the delivery room need to be expanded to reflect the physiologic and developmental care requirements. In this article, guidelines are proposed to address these needs. PMID- 12078319 TI - Life-sustaining support: ethical, cultural, and spiritual conflicts. Part II: Staff support--a neonatal case study. AB - As medical knowledge and technology continue to increase, so will the ability to provide life-sustaining support to patients who otherwise would not survive. Along with these advances comes the responsibility of not only meeting the clinical needs of our patients, but also of understanding how the family's culture and spirituality will affect their perception of the situation and their decision-making process. As the U.S. continues to become a more culturally diverse society, health care professionals will need to make changes in their practice to meet the psychosocial needs of their patients and respect their treatment decisions. Part I of this series (April 2002) discussed how the cultural and spiritual belief systems of Baby S's family affected their decision making processes and also their ability to cope with the impending death of their infant. The development of a culturally competent health care team can help bridge the gap between culturally diverse individuals. This article addresses the following questions: 1. What legal alternatives are available to the staff to protect the patient from suffering associated with the continuation of futile life-sustaining support? 2. What conflicts might the staff experience as a result of the continuation of futile life-sustaining support? 3. What efforts can be made to support members of the staff? 4. What can be done to prepare others in the health care professions to deal more effectively with ethical/cultural issues? PMID- 12078320 TI - A neonatal end-of-life palliative protocol--an evolving new standard of care? PMID- 12078321 TI - Direct hyperbilirubinemia: a case study of parenteral nutrition-induced cholestatic jaundice. PMID- 12078322 TI - Palliative care in the neonatal population. PMID- 12078323 TI - Neonatal glucose metabolism: differential diagnoses, evaluation, and treatment of hypoglycemia. AB - Hypoglycemia is one of the most common clinical care issues facing the neonatal practitioner. Increasing evidence indicates that neonatal hypoglycemia may have long-term neurologic effects. Care is complicated by the lack of a clearly defined threshold for hypoglycemia in term and preterm infants, however, and by highly variable clinical signs and symptoms. Furthermore, many infants with low blood glucose measurements do not exhibit obvious signs of impairment. The complexity of neonatal glucose metabolism is illustrated by the variety of conditions producing or associated with both neonatal hypo- and hyperglycemia. Maintenance of euglycemia is especially challenging in the sick or low birth weight neonate. This article defines euglycemia by its range and reviews the differential diagnoses and etiology of hypoglycemia--as well as the principles of its management--in the neonatal period. PMID- 12078324 TI - From human resource planning to human resource impact assessment: changing trends in health workforce strategies. PMID- 12078325 TI - The health manpower situation in Pakistan. PMID- 12078326 TI - Conflict, competition and cooperation: past and future developments of the professions allied to medicine in Australia. PMID- 12078327 TI - Issues when assessing future supply and requirements of medical workforce. PMID- 12078328 TI - The required number of physicians: is it an optimal figure? AB - Apart from the developing countries in which the number of physicians is extremely low, the number of physicians in the different countries shows a wide range even if it is comparing with the GDP or even with the number of nurses. According WHO data there are more than 500 physicians/100,000 inhabitants in Cuba and in Italy; more than 400 in Belarus and Norway, or in Spain and Georgia; more than 300 in 22 countries among them Belgium, France, Germany and Greece, Sweden, Switzerland, or Bulgaria, Czech, Hungary, Slovakia, Lithuania, etc. The number of doctors is below 300/100,000 inhabitants for example in such rich countries as the Netherlands, and the USA, and below 200 in the United Kingdom or in Japan. There is no correlation between the GDP and the number of doctors, as well as between the average life expectancy (or GBD, DALY, DALE) and the number of doctors. (On the other hand there is good correlation between GDP and life expectancy.) There is no correlation between GP-s and specialists, but it may be a weak correlation between the number of hospital beds and the number of doctors. It is difficult to find an optimal doctor/nurse ratio among the different countries; nevertheless the definition of "nurses" has a different meaning in the different countries. The burden of diseases, or the "patients" load" is also not an objective reference point, because the more the doctors, the more the patients. In the different countries there are different norms of the physicians' performances mainly decided by the health insurance funds. The different reimbursement systems, as the "per capita" at the GP-s, or the DRG at the hospital care, or the point-based remuneration at the specialist care are the real rate limiting factors for the number of physicians, which are not necessarily adequate to the professional requirements. PMID- 12078329 TI - [Human resources of the health system: status and evolution in industrialized countries]. AB - The physician-population ratio is high in the Mediterranean countries: nearly 4 doctors per 1000 population in Spain, more than 4 in Greece, nearly 6 in Italy. In Northern Europe, the ratio is lower. On the other side, as concerns nurses and other paramedical personnels, their ratio is much higher in the Scandinavian countries than in the South of Europe. Since 1945 onwards, all countries in Western Europe and Northern America have steadily increased the number of their health workers. The trend will no doubt continue during the decades ahead. The question is to what extent the current cost containment policies can decelerate this upward trend. As concerns physicians, the future panorama will be different. The "graduate-boom" cohorts of the 70's and 80's will reach their retirement age between 2010 and 2025 and will not be totally replaced. In other words, the (currently) plethoric medical profession will decrease after 2010 in most Western European countries, precisely at a time of growing population aging. PMID- 12078330 TI - Oral cytology in cannabis smokers. AB - The effects of cannabis/methaqualone/tobacco smoking on the epithelial cells of the tongue, buccal mucosa and floor of the mouth were examined. Oral mucosal smears for detection of cellular changes were taken from 4 sites in 16 patients. The tongue blade scraping technique was used. The sites sampled included the buccal mucosa (left and right sides), the posterior dorsum of the tongue and the anterior floor of the mouth. Tobacco smoking and non-smoking controls were also examined. The only significant difference between cannabis users and controls was the greater prevalence of bacterial cells in the smears taken from cannabis users. However, there were also greater numbers of degenerate and atypical squamous cells in cannabis smokers than in cigarette-smoking and non-smoking controls. Epithelial cells in smears taken from cannabis users and tobacco smoking controls also showed koilocytic changes, which were not seen in smears taken from non-smoking controls. Koilocytosis is indicative of human papilloma virus infection, although no apparent lesions were seen in the patients from whom smears had been taken. It would appear that there is a greater tendency towards damaged and immature surface epithelial cells in cannabis smokers. PMID- 12078331 TI - Follow-up study of apicectomised anterior teeth. AB - A retrospective study was carried out on anterior teeth which had undergone apicectomy between 1995 and 1997. More maxillary teeth were treated (94.7%) than mandibular teeth (5.3%). Radicular cyst was the most frequently diagnosed reason for the treatment (47.3%), followed by chronic apical infection/granuloma (44.0%). The overall success rate was 71.9%, while treatment outcome was doubtful in 14.0% of the apicectomised teeth. The presence of pre- or postoperative signs and symptoms did not have any statistically significant effect on the treatment outcome. A higher percentage of roots with retrograde amalgam fillings healed successfully than roots with orthograde fillings, but the difference was not statistically significant (P > 0.05). More lesions classified histopathologically as radicular cysts healed successfully than those classified as granulomas (Fisher exact test, 2-tailed P value = 1.0). PMID- 12078332 TI - Posterior composite resin restorations: Part I. Isolation. PMID- 12078333 TI - Some useful restorative materials and techniques in paedodontics. PMID- 12078334 TI - Dental record taking--what for (ensic)? Forensic communique. PMID- 12078335 TI - A novel slow-release formulation of lithium carbonate (Carbolithium Once-A-Day) vs. standard Carbolithium: a comparative pharmacokinetic study. AB - PURPOSE: The purpose of the study was to establish if the administration of a new slow-release formula of lithium carbonate (Carbothium Once-A-Day, 600 mg) administered once-daily could deliver plasma lithium levels during the first 24 hours similar to those obtained with two standard release Carbolithium 300 mg capsules administered 12 hours apart. PATIENTS AND METHODS: Eighteen healthy subjects of both sexes aged 18 to 55 were randomized to administration of either: [a] a single capsule of Carbolithium Once-A-Day (600 mg), or [b] standard Carbolithium 300 mg b.i.d. Subjects were crossed over following a 15-day washout period. Blood samples were taken 1, 2 4, 6, 8, 10, 12, 16, 20, 24, 36, 48, 72, and 96 hours post-drug administration and analyzed with spectrometry and atomic absorption to detect Li+ plasma concentration. RESULTS: Data for individual subjects are reported as disjointed numerical values and as individual graphs in the paper. Mean AUC values were 3.01 mM-hours for standard Carbolithium vs. 3.53 mM-hrs for Carbolithium Once-A-Day, and respective mean levels across 96 hours were 0.214 +/- 0.107 vs. 0.252 +/- 0.097 mM. For the first 24 hours, mean AUC values were 2.64 for Carbolithium vs. 3.03 mM-hours for Carbolithium Once-A-Day, and corresponding means were 0.264 vs. 0.303 mM. CONCLUSION: Carbolithium Once-A Day was associated with marked reduction of the peak/trough ratio compared to standard release Carbolithium. Given the low therapeutic index of lithium, the maintenance of constant therapeutic concentrations under toxic limits is an essential characteristic of any clinically useful formulation. Furthermore, from the data obtained in the present study it is predicted that, for the majority of patients, a single dose of Carbolithium Once-A-Day will be sufficient to provide therapeutic concentrations of lithium for 24-hour periods. Even the few subjects who will require a double dosage with the Once-A-Day formulation will certainly experience less variations of Li+ plasma concentration throughout the day than would patients receiving rapid release formulations of lithium. PMID- 12078336 TI - [Women and the mystery of autoimmune diseases]. PMID- 12078337 TI - [Clinical investigation of hyaluronidase and nerve growth factor (NGF) at the ocular level: detailed review of the international literature]. AB - In this paper the authors analyse the international bibliography about clinic tests on hyaluronic acid and Nerve Growth Factor at ocular level. This study was performed on request of a private oculistic clinic in the centre of Rome about the clinic test of the drugs mentioned above. Therefore, this paper is a joint work of Public Health physicians and oculists. PMID- 12078338 TI - [Prevention and treatment of mucositis from cytotoxic chemotherapy]. AB - The administration of several chemotherapeutic regimens could be conditioned by the onset of mucositis. The characteristic lesions of the mucositis affect whole buccal mucosa. That derives from rapid turnover of the oropharyngeal epithelial surfaces. The mucosa can suffer from direct damage of antiblastic drugs or be susceptible of microbic infections. Moreover, other factors correlated to the patients as age, nutritional status, tumor type, oral hygiene and neutrophil count. Up to date, there is not a standard therapy for the cure or mucositis prevention. Some formalities can be employed in order to reduce chemo-induced damage: 1) altering the distribution and the excretion of drugs on the mucosa; 2) stimulating the basal cells of the mucosa; 3) trying to modify the infectious or inflammatory risks. The effective oral care, dietary changes and the use of protective topical and the careful use of topical and systemic anesthetic drugs are the cornerstones of mucositis care. PMID- 12078340 TI - [25 years old woman with nocturia]. AB - A 25 years old lady presented to our outpatient clinic complaining nicturia, polyuria and polydipsia. On the basis of clinical assessment and the results of a simple test, a diagnosis of Nephrogenic Diabetes Insipidus (NDI) was made. The diagnosis of NDI is often missed, and appropriate treatment therefore delayed, with severe consequences especially in elderly and bed restricted people. PMID- 12078339 TI - [Treatment of ovarian cancer]. AB - PURPOSE: To present an approach to the treatment of epithelial ovarian cancer. DESIGN: A large number of studies in ovarian cancer have been reviewed. RESULTS: Ovarian cancer is the most common fatal malignancy of the female genital tract. The high mortality is due to the fact that the most part of patients are diagnosed with advanced disease. Cytoreductive surgery followed by platinum-based chemotherapy is the standard therapeutic strategy for patients with advanced ovarian cancer. Postsurgical residual disease is the most important prognostic factor. The combination of Paclitaxel and Cisplatin or Carboplatin is currently accepted as the standard chemotherapy for advanced ovarian cancer. Second- look laparatomy should be reserved to patients enrolled in clinical trial. Resistant disease remain the major cause of relapse and death. Several drugs have been employed in second-line therapy; however, the responses are generally short and survival is poor. CONCLUSIONS: Future research should include new drugs or new combination regimens to improve clinical outcomes and overall survival. PMID- 12078341 TI - [To the new Family Medicine Rubric]. PMID- 12078342 TI - [Urinary tract infections]. PMID- 12078343 TI - [Non-traumatic rhabdomyolysis: clinical and laboratory diagnosis, assessment of renal complications]. PMID- 12078344 TI - [27th National Oncology Congress of the Italian Tumor Society. Part 2]. PMID- 12078345 TI - [Survey of five years psychiatric consultations in a university general hospital]. AB - A survey of 5 year period activity (1995-99) of a university consultation-liaison service is reported, concerning 1556 psychiatric consultations. The majority of referrals (68%) came from internal medicine units; the reason of referral was prevalently due (approx. 50%) to the presence of psychiatric symptoms; moreover, the majority of patients (approx. 51%) for which the consultation has been requested were already under psychiatric treatment and 32% of them took psychoactive drugs. In 75% of cases a psychiatric diagnosis according to ICD-10 was formulated after consultation: "Affective syndromes" were diagnosed in 25% of cases and "Neurotic, somatoformand stress-related syndromes" in 36.6% of cases. Approx. 60% of patients were prescribed psychoactive drugs; 50% received psychological support. PMID- 12078346 TI - Age-related trend in circadian rhythm of electrocardiographic sinusal R-R intervals in clinically healthy subjects. AB - It is a clinical observation that human HR shows a tendency to bradycardia with advancing age. In addition, there is experimental evidence that heart rate (HR) shows a circadian rhythm (CR) in healthy humans. Because of this background, the present study is devoted to investigate how the CR of the electrocardiographic sinusal R-R intervals (SRRI) changes its properties as a function of chronological age in healthy humans. The study was performed on 170 clinically healthy Japanese, ranging in age from 12 to 83 years (mean age +/- SD = 43 +/- 15 years), who were monitored in their electrocardiogram over the day-night span. An opposite age-related trend was detected for two properties of SRRI CR, i.e., mesor (positive trend) and amplitude (negative trend). Accordingly, the SRRI CR can be classified as an "amphiclinous rhythm", i.e., a rhythm in which the cardiac interbeat duration shows a progressive increase of its daily mean value along with a progressive deamplification of its oscillatory extent. Furthermore, the rhythm is characterized by a progressive anticipation in time of its oscillatory maximum of duration with advancing age. The findings suggest that the biological clocks regulating the HR CR in human beings undergo a resetting of their mechanisms of tonic, amplitude and phasic modulation as a function of age. The clock-mediated readjustment of the HR CR may be regarded as one of the mechanisms for explaining the bradycardia that is associated with the aging process in human beings. PMID- 12078348 TI - Diabetes. Complementary and alternative care. PMID- 12078347 TI - [Technology assessment and operating procedures for the technological resources in the new Community Health Service]. AB - INTRODUCTION: The main purpose of this contribution is to set today's situation of technology assessment and point to some organizing and integrative ways with the evolution concern ASL (Local Health Firm). PATIENTS AND METHODS: We outline main critical processes that fix correct management of technological resource thought data available in national and international literature. We single out more common methods to quantify, evaluate, control and manage biomedical technologies. RESULTS: We expound decisional process that lead Viterbo ASL (Local Health Firm) to a progressive controll of tecnological resource and its management after aggregation of the five USL (Local Health Units) that constituted Viterbo ASL and describe procedures assumed. CONCLUSIONS: We underline the importance of full and complete information in business and decisional process. Inside the complexity of italian view, we outline an organizative solution with a low economic impact and with a high technical content. PMID- 12078349 TI - Recommendations for herbal supplements and the perioperative patient. PMID- 12078350 TI - Reflexology. PMID- 12078351 TI - Comparing apples with apples in clinical populations: applications of the adjusted Clinical Group System in British Columbia. AB - This article reviews the Adjusted Clinical Group Case-Mix System and describes how it is being applied in the management of physician services in British Columbia. Developed in the United States for management and research, adjusted clinical groups are used to measure the illness burden and health service needs of individuals and, when aggregated, of populations, by grouping the range of conditions coded on physician claims and hospital care records over a defined time period, typically one year. In Canadian and United States settings, adjusted clinical groups are up to five times more predictive of ambulatory resource use than are age and sex groups alone. The article describes how adjusted clinical groups are being applied to adjust capitation payments for physician groups in British Columbia's Primary Care Demonstration Project and profiles of physician practice activity. PMID- 12078352 TI - Organizational learning and continuous quality improvement: examining the impact on nursing home performance. AB - Interest is growing in learning more about the ability of total quality management and continuous quality improvement (TQM/CQI) initiatives to contribute to the performance of healthcare organizations. A major factor in the successful implementation of TQM/CQI is the seminal contribution of an organization's culture. Many implementation efforts have not succeeded because of a corporate culture that failed to stress broader organizational learning. This may help to explain why some TQM/CQI programs have been unsuccessful in improving healthcare organization performance. Organizational performance variables and organizational learning orientation were assessed in a sample of 181 Canadian long-term care organizations that had implemented a formal TQM/CQI program. Categorical regression analysis shows that, in the absence of a strong corporAte culture that stresses organizational learning and employee development, few performance enhancements are reported. The results of the assessment suggest that a TQM/CQI program without the backing of a strong organizational learning culture may be insufficient to achieve augmented organizational performance. PMID- 12078353 TI - Toward an accountability framework for Canadian healthcare. AB - State-funded healthcare systems increasingly recognize accountability as an important public policy issue. This article explores significant aspects of current theory and practice in order to describe an accountability framework for the Canadian health system. Stakeholders include governments, institutions, providers and patients. Their relationships may be framed in constitutional, political, financial, managerial, clinical or ethical terms. The specific processes and instruments to operationalize accountability depend on the terms by which it is framed. PMID- 12078354 TI - What can the American electronic health record (EHR) pioneers tell us about what it takes to be successful? PMID- 12078355 TI - Be a life-long learner. PMID- 12078356 TI - 10 steps to a successful governance transfer. AB - The governance of several provincial psychiatric hospitals was transferred to the public hospitals across Ontario, with London and St. Thomas being two of them There were many learnings gained from the London/St. Thomas experience that can be taken forward to other transfers such as this. The keys to success included having a central negotiating table across all receiving hospitals, developing principles or values for a common vision across all parties, effective communication and commitment to the process. These learnings will be brought forward into Tier 2 and 3 of the transfers, when beds and programs will decentralize across Southwestern Ontario, and a reinvestment in community mental health will support the de-institutionalization of mental healthcare. PMID- 12078357 TI - Connecting for change: networks as a vehicle for regional health reform the early experiences of the Child Health Network for the Greater Toronto Area. AB - The Child Health Network (CHN) for the Greater Toronto Area (GTA) is a partnership of hospital, rehabilitation and community providers committed to developing a regional system to deliver high quality, accessible, family-centred care for mothers, newborns, children and youth. This article reviews the history and model of the CHN, assesses its achievements, and provides insights into the challenges and lessons learned by the network. Stemming from the CHN's commitment to quality, accessibility and efficiency, regionalization of maternal, newborn and children's services is emerging as a success story. PMID- 12078358 TI - Measuring and managing waiting times: what's to be done? AB - On September 11, 2000, the First Ministers of Canada issued a communique pledging to develop and report on waiting times for a number of diagnostic and treatment services. Reporting is to begin by September 2002. Given this commitment, what are the ideal characteristics of such a data collection system? This article defines and evaluates methods of measuring waiting times, and recommends a prioritized waiting-time information system to permit both measurement and management. PMID- 12078359 TI - Quality and safety in health care: a time of transition. PMID- 12078360 TI - Ernest Amory Codman MD. PMID- 12078361 TI - Getting into the red: a strategic step for safety. PMID- 12078362 TI - Barriers to incident reporting in a healthcare system. AB - BACKGROUND: Learning from mistakes is key to maintaining and improving the quality of care in the NHS. This study investigates the willingness of healthcare professionals to report the mistakes of others. METHODS: The questionnaire used in this research included nine short scenarios describing either a violation of a protocol, compliance with a protocol, or improvisation (where no protocol exists). By developing different versions of the questionnaire, each scenario was presented with a good, poor, or bad outcome for the patient. The participants (n = 315) were doctors, nurses, and midwives from three English NHS trusts who volunteered to take part in the study and represented 53% of those originally contacted. Participants were asked to indicate how likely they were to report the incident described in each scenario to a senior member of staff. RESULTS: The findings of this study suggest that healthcare professionals, particularly doctors, are reluctant to report adverse events to a superior. The results show that healthcare professionals, as might be expected, are most likely to report an incident to a colleague when things go wrong (F(2,520) = 82.01, p < 0.001). The reporting of incidents to a senior member of staff is also more likely, irrespective of outcome for the patient, when the incident involves the violation of a protocol (F(2,520) = 198.77, p < 0.001. It appears that, although the reporting of an incident to a senior member of staff is generally not very likely, particularly among doctors, it is most likely when the incident represents the violation of a protocol with a bad outcome. CONCLUSIONS: An alternative means of organisational learning that relies on the identification of system (latent) failures before, rather than after, an adverse event is proposed. PMID- 12078364 TI - Shame: the elephant in the room. PMID- 12078363 TI - Implementing a new drug record system: a qualitative study of difficulties perceived by physicians and nurses. AB - OBJECTIVES: To identify organisational difficulties faced by physicians and nurses when using drug prescribing sheets for recording both drug prescriptions and drug administration. DESIGN: Qualitative interview study. SETTING: Two general internal medicine wards. PARTICIPANTS: Seven physicians and eight nurses. MAIN OUTCOME MEASURES: Difficulties explicitly identified by the participants during the interviews. RESULTS: The implementation of procedures conflicted with existing structure, culture, and routines. Insufficient competence within the system to use the drug prescribing sheets created resistance and made people down the line create their own interpretations and solutions to the problems they faced. A total of nine problems were identified: (1) insufficient knowledge and uncertainty about procedures, (2) ignorance of sources of error, (3) unclear responsibilities, (4) low community spirit, (5) insufficient communication, (6) clinician autonomy and low acceptance of change, (7) strong professional identity, (8) low priority task, and (9) logistical problems. CONCLUSIONS: Unawareness of procedures, insufficient dissemination of knowledge, and insufficient cooperation and skepticism among those who put drug handling into practice is likely to have an impact on the quality of health care. The identification of these obstacles may help managers to improve the quality of the drug handling process on internal medicine wards and make it possible to select a framework for changing the clinical behaviour of doctors and nurses. PMID- 12078366 TI - The end of the beginning: the strategic approach to patient safety research. PMID- 12078365 TI - Development and description of a decision analysis based decision support tool for stroke prevention in atrial fibrillation. AB - BACKGROUND: There is an increasing move towards clinical decision making that engages the patient, which has led to the development and use of decision aids to support better decisions. The treatment of patients in atrial fibrillation (AF) with warfarin to prevent stroke is a decision that is sensitive to patient preferences as shown by a previous decision analysis. AIM: To develop a computerised decision support tool, building upon a previous decision analysis, which would engage individual patient preferences in reaching a shared decision on whether to take warfarin to prevent stroke. METHODS: The development process had two main phases: (1) the development phase which employed focus groups and repeated interviews with GPs/practice nurses and patients alongside an iterative development of a computerised tool; (2) the training and testing phase in which GPs and practice nurses underwent training in the use of the tool, including the use of simulated patients. The tool was then used in a feasibility study in a small number of patients with AF to inform the design of a subsequent randomised controlled trial. RESULTS: The prototype tool had three components: (1) derivation of an individual patient's values for relevant health states using a standard gamble; (2) presentation/discussion of a patient's risks of stroke using the Framingham equation and the benefits/risks of warfarin from a systematic literature review; and (3) decision making component incorporating the outcome of a Markov decision analysis model. Older patients could be taken through the decision analysis based computerised tool, and patients and clinicians welcomed information on risks and benefits of treatments. The tool required time and training to use. Patients' decisions in the feasibility phase did not necessarily coincide with the output of the decision analysis model, but decision conflict appeared to be reduced and both patients and GPs were satisfied with the process. CONCLUSIONS: It is feasible to develop a decision analysis based computer software package that is acceptable to elderly patients and clinicians, but it requires time and expertise to use. It is most likely that a tool of this type will best be used by a small number of clinicians who have developed experience of its use and can maintain their skills. PMID- 12078367 TI - Development and preliminary evaluation of a clinical guidance programme for the decision about prophylactic oophorectomy in women undergoing a hysterectomy. AB - OBJECTIVES: To develop a decision analysis based and computerised clinical guidance programme (CGP) that provides patient specific guidance on the decision whether or not to undergo a prophylactic oophorectomy to reduce the risk of subsequent ovarian cancer and to undertake a preliminary pilot and evaluation. SUBJECTS: Women who had already agreed to have a hysterectomy who otherwise had no ovarian pathology. SETTING: Oophorectomy decision consultation at the outpatient or pre-admission clinic. METHODS: A CGP was developed with advice from gynaecologists and patient groups, incorporating a set of Markov models within a decision analytical framework to evaluate the benefits of undergoing a prophylactic oophorectomy or not on the basis of quality adjusted life expectancy, life expectancy, and for varying durations of hormone replacement therapy. Sensitivity analysis and preliminary testing of the CGP were undertaken to compare its overall performance with established guidelines and practice. A small convenience sample of women invited to use the CGP were interviewed, the interviews were taped and transcribed, and a thematic analysis was undertaken. RESULTS: The run time of the programme was 20 minutes, depending on the use of opt outs to default values. The CGP functioned well in preliminary testing. Women were able to use the programme and expressed overall satisfaction with it. Some had reservations about the computerised formal and some were surprised at the specificity of the guidance given. CONCLUSIONS: A CGP can be developed for a complex healthcare decision. It can give evidence-based health guidance which can be adjusted to account for individual risk factors and reflects a patient's own values and preferences concerning health outcomes. Future decision aids and support systems need to be developed and evaluated in a way which takes account of the variation in patients' preferences for inclusion in the decision making process. PMID- 12078368 TI - Combating omission errors through task analysis and good reminders. AB - Leaving out necessary task steps is the single most common human error type. Certain task steps possess characteristics that are more likely to provoke omissions than others, and can be identified in advance. The paper reports two studies. The first, involving a simple photocopier, established that failing to remove the last page of the original is the commonest omission. This step possesses four distinct error-provoking features that combine their effects in an additive fashion. The second study examined the degree to which everyday memory aids satisfy five features of a good reminder: conspicuity, contiguity, content, context, and countability. A close correspondence was found between the percentage use of strategies and the degree to which they satisfied these five criteria. A three stage omission management programme was outlined: task analysis (identifying discrete task steps) of some safety critical activity; assessing the omission likelihood of each step; and the choice and application of a suitable reminder. Such a programme is applicable to a variety of healthcare procedures. PMID- 12078369 TI - Improving safety on the front lines: the role of clinical microsystems. AB - The clinical microsystem puts medical error and harm reduction into the broader context of safety and quality of care by providing a framework to assess and evaluate the structure, process, and outcomes of care. Eight characteristics of clinical microsystems emerged from a qualitative analysis of interviews with representatives from 43 microsystems across North America. These characteristics were used to develop a tool for assessing the function of microsystems. Further research is needed to assess microsystem performance, outcomes, and safety, and how to replicate "best practices" in other settings. PMID- 12078370 TI - Leading organisational learning in health care. AB - As healthcare organisations seek to enhance safety and quality in a changing environment, organisational learning practices can help to improve existing skills and knowledge and provide opportunities to discover better ways of working together. Leadership at executive, middle management, and local levels is needed to create a sense of shared purpose. This shared vision should help to build effective relationships, facilitate connections between action and reflection, and strengthen the desirable elements of the healthcare culture while modifying outdated assumptions, procedures, and structures. PMID- 12078371 TI - Pushing the profession: how the news media turned patient safety into a priority. AB - The problem of patient safety has been repeatedly identified in the medical literature since the mid 1950s, but regular revelations about patient deaths and injuries resulting from treatment have had almost no effect on the actual practice of medicine. Only very recently has the medical profession made a systematic effort to reduce or eliminate the many preventable deaths and injuries that occur in hospitals each year. This review traces the diffusion of innovation in medical error reduction to the public shaming of the profession that occurred as a result of stories that appeared in the news media. The focus is on the USA, but news stories about patient safety are sparking a similar process throughout the western world. PMID- 12078372 TI - Why don't physicians enthusiastically support quality improvement programmes? PMID- 12078373 TI - A system of medical error disclosure. AB - External mandates for medical error disclosure are often justified by potential cost savings, the belief in individual moral obligations in health care, and the concept that patients have rights and providers have responsibilities. Such an approach does not recognise the systems nature of error and outcomes and the important quality role disclosure can play in a system of medical error disclosure. Systems concepts, the patient-provider partnership, and overall quality of care can be enhanced using a system of disclosure that provides for education about the systems nature of error, fulfills the delivery system philosophy of mutual respect, and integrates the patient and his/her family as a partner in the error reduction enterprise. Such a system can result using clear disclosure policies and procedures sensitive to patient and family needs, open communications with concerned, committed, and compassionate system representatives, and use of mediation methods that foster communication, allow for venting, and are flexible in their approach to resolving conflict, including using apology. Although a system may also result in conflict resolution costs, more importantly it may foster and solidify a team approach to reducing errors and promoting patient safety. PMID- 12078375 TI - Telling patients the truth: a systems approach to disclosing adverse events. PMID- 12078376 TI - Patient safety: what about the patient? AB - Plans for improving safety in medical care often ignore the patient's perspective. The active role of patients in their care should be recognised and encouraged. Patients have a key role to play in helping to reach an accurate diagnosis, in deciding about appropriate treatment, in choosing an experienced and safe provider, in ensuring that treatment is appropriately administered, monitored and adhered to, and in identifying adverse events and taking appropriate action. They may experience considerable psychological trauma both as a result of an adverse outcome and through the way the incident is managed. If a medical injury occurs it is important to listen to the patient and/or the family, acknowledge the damage, give an honest and open explanation and an apology, ask about emotional trauma and anxieties about future treatment, and provide practical and financial help quickly. PMID- 12078374 TI - Decision support and safety of clinical environments. AB - Safety in the clinical environment is based on structures that reduce the probability of harm, on evidence that enhances the likelihood of actions that increase favourable outcomes, and on explicit directions that lead to decisions to implement the actions dictated by this evidence. A clinical decision error rate of only 1% threatens patient safety at a distressing frequency. Explicit computerised decision support tools standardise clinical decision making and lead different clinicians to the same set of diagnostic or therapeutic instructions. They have favourable impacts on patient outcome. Simple computerised algorithms that generate reminders, alerts, or other information, and protocols that incorporate more complex rules reduce the clinical decision error rate. Decision support tools are not new; it is the new attributes of explicit computerised decision support tools that deserve identification. When explicit computerised protocols are driven by patient data, the protocol output (instructions) is patient specific, thus preserving individualized treatment while standardising clinical decisions. The expected decrease in variation and increase in compliance with evidence-based recommendations should decrease the error rate and enhance patient safety. PMID- 12078377 TI - Should we consider non-compliance a medical error? AB - Non-compliance is an extensive intractable problem. This paper argues that we can gain significant insight into non-compliance if we apply theories developed to explain human error in organisations. The resultant framework encompasses intentional and unintentional non-compliance, shifts blame from the patient, and recognises the influence of other factors, including organisational ones. There are also consequences for the measurement of compliance and new strategies to improve it. Terminology will need to be addressed, particularly whether intentional non-compliance by a patient should be considered an error. If empirical research supports the arguments in this paper then, with some further theory development, the application of human error theory will offer a useful new approach to understanding and reducing undesired non-compliance. PMID- 12078378 TI - Effectiveness of quality improvement: learning from evaluations. AB - The effectiveness of many quality improvement interventions has been studied, and research suggests that most have highly variable effects which depend heavily on the context in which they are used and the way they are implemented. This has three important implications. Firstly, it means that the approach to quality improvement used in an organisation probably matters less than how and by whom it is used. Rather than taking up, trying, and then discarding a succession of different quality improvement techniques, organisations should probably choose one carefully and then persevere to make it work. Secondly, future research into quality improvement interventions should be directed more at understanding how and why they work--the determinants of effectiveness--rather than measuring whether they work. Thirdly, some element of evaluation should be incorporated into every quality improvement programme so that its effectiveness can be monitored and the information can be used to improve the systems for improvement. PMID- 12078379 TI - Quality in health care: a role for the law? AB - This paper considers the way in which English law can and does play a part in ensuring quality standards in healthcare practice. It examines the enhanced scope for judicial scrutiny of standards of care and the role played by law in the form of statutory regulation. There is an increasing degree of legal involvement in healthcare decision making. The law may provide some effective checks and balances to quality but, at the same time, the limits of the law as an effective regulator of quality have to be recognised because of the multifaceted nature of clinical judgment. PMID- 12078380 TI - Implementing clinical governance in English primary care groups/trusts: reconciling quality improvement and quality assurance. AB - OBJECTIVES: To investigate the concept of clinical governance being advocated by primary care groups/trusts (PCG/Ts), approaches being used to implement clinical governance, and potential barriers to its successful implementation in primary care. DESIGN: Qualitative case studies using semi-structured interviews and documentation review. SETTING: Twelve purposively sampled PCG/Ts in England. PARTICIPANTS: Fifty senior staff including chief executives, clinical governance leads, mental health leads, and lay board members. MAIN OUTCOME MEASURES: Participants' perceptions of the role of clinical governance in PCG/Ts. RESULTS: PCG/Ts recognise that the successful implementation of clinical governance in general practice will require cultural as well as organisational changes, and the support of practices. They are focusing their energies on supporting practices and getting them involved in quality improvement activities. These activities include, but move beyond, conventional approaches to quality assessment (audit, incentives) to incorporate approaches which emphasise corporate and shared learning. PCG/Ts are also engaged in setting up systems for monitoring quality and for dealing with poor performance. Barriers include structural barriers (weak contractual levers to influence general practices), resource barriers (perceived lack of staff or money), and cultural barriers (suspicion by practice staff or problems overcoming the perceived blame culture associated with quality assessment). CONCLUSION: PCG/Ts are focusing on setting up systems for implementing clinical governance which seek to emphasise developmental and supportive approaches which will engage health professionals. Progress is intentionally incremental but formidable challenges lie ahead, not least reconciling the dual role of supporting practices while monitoring (and dealing with poor) performance. PMID- 12078382 TI - Births, marriages, divorces, and deaths: provisional data for January-March 2001. PMID- 12078383 TI - Dracunculiasis eradication. Khartoum declaration on guinea-worm eradication, Sudan, 2002. PMID- 12078384 TI - Natural or raw almonds and an outbreak of a rare phage type of Salmonella enteritidis infection. PMID- 12078385 TI - Revision of the International Health Regulations--progress report, May 2002. PMID- 12078381 TI - Acupuncture. PMID- 12078386 TI - [Effect of vitamin C on platelet aggregation in smokers and nonsmokers]. AB - BACKGROUND: Epidemiologic studies suggest a protective effect of regular intake of vitamin C and vitamin E as antioxidant in the manifestation of coronary heart disease. Cigarette smoke contains a large amount of radicals and reactive oxygen derived substances enhancing aggregation of platelets. We investigated the effect of vitamin C as an important antioxidant in human plasma on the aggregation of human platelets in smokers and nonsmokers. TEST PERSONS AND METHOD: Overall 40 persons (mean age: 28 +/- 9 years) were randomized. The groups of chronic smokers (21 +/- 9 "packyears") and nonsmokers consisted of 20 persons, respectively. In each group ten persons were treated with intravenous infusion of 3 g vitamin C or 100 ml 0.9% saline solution (placebo). The maximal aggregation was measured with an aggregometer after 0, 3, 6, and 24 hours with collagen concentrations of 0.5 microgram/ml and 1.0 microgram/ml, respectively. RESULTS: In smokers with vitamin C application the group comparison by Wilcoxon's rank test demonstrated a significant decrease of platelet aggregation after 6 hours for both collagen concentrations (0.5 microgram/ml and 1.0 microgram/ml) compared to the placebo group (p < or = 0.05), whereas nonsmokers with vitamin C application revealed a significant decrease of platelet aggregation after 3 and 6 hours for both collagen concentrations (0.5 microgram/ml and 1.0 microgram/ml) compared to the placebo group (p < or = 0.03). The comparison between smokers and nonsmokers regarding the effect of vitamin C on platelet aggregation for both collagen concentrations demonstrated no significant difference (3 hours: p = 0.84 and p = 0.97; 6 hours: p = 0.81 and p = 0.59; and 24 hours p = 0.57 and p = 0.06, not significant, respectively). CONCLUSION: These findings suggest that vitamin C exerts an unknown inhibitory effect on collagen-induced platelets aggregation. These observations may represent a further protective effect of vitamin C in the development of coronary heart disease. PMID- 12078387 TI - [Deficits in management of patients with colorectal carcinoma in Germany. Results of multicenter documentation of therapy algorithms]. AB - BACKGROUND: Adjuvant chemotherapy for patients with UICC III (Dukes C) colorectal cancer (consensus statements NIH 1990, German Cancer Society 1994) and palliative chemotherapy for metastatic disease have long been recognized to provide a survival benefit in colorectal cancer. However, it remains unclear if these concepts have made inroads into clinical practice. PATIENTS AND METHODS: Therefore, we asked 74 institutions treating colorectal cancer patients in Germany to document the treatment algorithms of all patients with colorectal cancer seen in the third quarter of 1998. Clinical careers of 1,001 patients (m/f 465/536; median age 62.9 years [28-93]; colon 596, rectum 405; UICC I 117, II 206, III 407, IV 218) were documented. RESULTS: Only 63.4% of patients with UICC III colorectal cancer received adjuvant therapy with a significant difference between hospitals with (67.1%) and without (42.6%) oncological departments (p < 0.01). Higher age appeared to be the most important factor for withholding treatment since 196 of 286 (68.5%) patients < 70 years, but only 57 of 121 (47.1%) > 70 years underwent adjuvant therapy. 78.4% of patients with UICC IV colorectal cancer (91.8% university hospitals, 76.8% hospital with, 50% without oncological departments, 66.7% rehabilitation clinics, 82.4% private practices) received palliative chemotherapy (first line: 5-FU/FA bolus 57%, 5-FU/FA infusion 20%, 5-FU mono 15%). CONCLUSION: Considering an annual incidence of colorectal cancer in Germany of 52,000 with 30% UICC III, discounting patients > 80 years or ECOG status > 2, and estimating a survival benefit of 10% after adjuvant chemotherapy, approximately 530 lifes are lost annually in Germany due to insufficient treatment of UICC III colorectal cancer based on our survey. In addition, substantial financial demand is generated by the subsequent palliative treatment of potentially curable patients.--In conclusion, survey-based analysis of treatment algorithms can provide valuable insights into clinical practice in oncology and can disclose deficits in patient care as demonstrated here in colorectal cancer. PMID- 12078388 TI - [Genetic risk factors in pancreatic diseases--significance for general practice]. AB - GENETIC RISK FACTORS: Recently, several genetic risk factors for chronic pancreatic diseases were found. In patients with chronic pancreatitis several mutations of the cationic trypsinogen were identified. In the majority of these subjects an autosomal dominant disease was observed. Mutations in the pancreatic secretory trypsin inhibitor (SPINK 1) were found in 20% of subjects with idiopathic, in 5% of those with alcoholic chronic and in 50% of those with tropical pancreatitis. Further variants were identified in CFTR (Cystic Fibrosis Transmembrane Conductance Regulator), a chlorid transporter initially identified a disease-causing molecule in cystic fibrosis. In approximately 20-25% of the patients with chronic pancreatitis a mutation of one of these genes can be found. INDICATION FOR EXAMINATION: A genetic investigation is useful when there is evidence for a family history of chronic pancreatitis and, at absence of typical risk factors, in patients with onset of disease earlier than 35-40 years of age. RECOMMENDATIONS: In addition to alcohol abstinence the patients should be advised not to smoke. National and international registers as well as self-aid groups exist. PMID- 12078389 TI - [Arterial vascular screening with whole body MR angiography]. AB - Atherosclerosis represents a frequent health problem in developed countries. Although a systemic disease, the diagnostic approach to atherosclerosis has remained segmental. Largely, this approach has been a reflection of limitations inherent to the imaging techniques used. Recently, whole-body MR angiography has become possible allowing the display of the arterial vasculature from the supraaortic arteries to the distal run-off vessels in merely 72 seconds. The technique provides images quality comparable to catheter angiography. The advantage of this approach relates to its potential in detecting vascular pathologies in regions which would have remained unexamined with conventional diagnostic management. Thus, the outlined strategy of whole-body MR angiography mirrors the systemic nature of atherosclerosis and can be used as a noninvasive comprehensive screen for arterial disease screening. PMID- 12078390 TI - [Biological therapies for treatment of chronic inflammatory bowel diseases. Status update]. AB - BACKGROUND: For the treatment of inflammatory bowel diseases in recent years several antibodies, cytokines and antisense oligonucleotides have been developed using recombinant technology and were tested as so-called "biological therapeutics". EFFECTIVENESS OF BIOLOGICAL THERAPEUTICS: Infliximab, a chimeric TNF-alpha antibody, is the only biological remedy approved for the treatment of refractory and fistulizing Crohn's disease. Because of inherent risks for severe side effects, such as sepsis, the indication should be restricted to truly refractory patients and treated patients must be followed very carefully. In clinical trials other anti-TNF-alpha-effective substances (CDP 571, etanercept, thalidomide), interleukin-10, interleukin-11, ICAM-1-antisense-oligonucleotides and antibodies against alpha 4-integrin were evaluated for the treatment of Crohn's disease. In summary, for all a slight effect was noted but their place in the therapeutical repertoire is not yet defined. None of these substances is approved for patients with inflammatory bowel disease. Future developments are eagerly awaited. PMID- 12078391 TI - [Acute symptomatic hyponatremia in a 70-year-old patient. Insufficient ADH secretion syndrome]. AB - CASE REPORT: A 70-year-old male was admitted to the hospital with anorexia, nausea, headache, and apathy. Serologically remarkable were a severe hyponatremia, serum hypoosmolality and urine hyperosmolality with normal renal function. After excluding adrenal insufficiency, hypothyroidism and other possible causes of hypoosmolar hyponatremia, the diagnosis of a syndrome of inadequate ADH secretion (SIADH) was confirmed through a nonsuppressed ADH level and a rise in of serum sodium after fluid restriction. Magnetic resonance imaging showed a small, chronic, traumatic subdural hematoma which was responsible for the SIADH in our patient. CONCLUSION: This etiologic correlation was reported in literature by many authors. However, these SIADHs were associated with a big subdural hematoma which needed neurosurgical intervention in most of the cases. Our patient is unique in that his symptoms were caused by a small subdural hematoma, which did not need surgical intervention. The patient was treated with cautious saline infusion, administration of a loop diuretic and fluid restriction with improvement of both his clinical and metabolic state. As large water loads may result in severe hyponatremia in these patients, moderate fluid restriction appears to be effective in long-term management even after stabilization/normalization of serum sodium. There are no sufficient data about the incidence of asymptomatic hyponatremia due to SIADH resulting from minor head injury, because most of these patients are asymptomatic and therefore they do not receive clinical attention. PMID- 12078392 TI - [Pulmonary alveolar microlithiasis--a rare cause of bilateral extensive pulmonary infiltrates]. AB - BACKGROUND: Pulmonary alveolar microlithiasis is a rare disease, which is characterized by pulmonary deposition of calcium phosphate microliths. The radiographic features can be pathognomonic with a "sandstorm"-like opacification throughout the lungs. CASE REPORT: A 29-year-old Moroccan presented for the first time 1988 with atypical chest pain. His history was characterized by nephrocalcinosis and pulmonary disease. Physical examination and laboratory values were unremarkable. Chest X-ray revealed extensive bilateral pulmonary infiltrates with "sandstorm"-like opacifications. Sonography showed medullary nephrocalcinosis. The diagnosis of pulmonary alveolar microlithiasis was confirmed by transbronchial biopsy. Over the next 13 years there was only a moderate expansion of the pulmonary infiltrates and a beginning restrictive pattern, whereas the patient was almost symptom-free. CONCLUSION: Pulmonary alveolar microlithiasis is a rare cause of bilateral pulmonary infiltrates and should be considered in the differential diagnosis. In some cases there is also an association with calcifications of extrapulmonary organs. PMID- 12078393 TI - [Absolute arrhythmia and pneumatosis cystoides intestinalis]. PMID- 12078395 TI - Merger creates new staff blend. PMID- 12078394 TI - [Organization development of the public health system]. AB - BACKGROUND: Changes in the German health care system require changes in health care institutions. Organizational development (OD) techniques can help them to cope successfully with their changing environment. CONCEPT: OD is defined as a collective process of learning aiming to induce intended organizational change. OD is based on social science methods and conducted by process-oriented consultants. In contrast to techniques of organizational design, OD is characterized by employee participation. One of the most important elements of OD is the so-called "survey-feedback-technique". EXAMPLES: Five examples illustrate how the survey-feedback-technique can be used to facilitate organisational learning. CONCLUSIONS: OD technique supports necessary change in health care organizations. It should be used more frequently. PMID- 12078396 TI - The CFO's role in software selection. PMID- 12078397 TI - Managed care applications evolve. PMID- 12078398 TI - Electronic medical records: a 'workflow' in progress. PMID- 12078399 TI - Not your father's benefits package. PMID- 12078400 TI - Reader's perspectives. Personal digital assistants (PDAs) will greatly improve data collection... . PMID- 12078401 TI - Front-line leadership. Chapter 1: Blonde ambition. PMID- 12078402 TI - The Golden Hour: fact or fiction? PMID- 12078403 TI - An EMS system for India. PMID- 12078406 TI - The silent worker. AB - The patient in this case was diagnosed as having a myocardial infarction, exacerbated over time. It has been reported in medical literature that women in such cases may have atypical symptoms that mimic those of other medical conditions and deceive providers into misdiagnosis. A few women present with the classical chest pain, a symptom most people relate to a feeling of a heart attack. This symptom is more typical in males. EMS providers should have a high suspicion of heart disease in any patient who describes their symptoms as common or relative to acute coronary syndromes. Patients at risk include those with high blood pressure, coronary artery disease, increased cholesterol, obesity and diabetes, among others. Given that this patient was resting comfortably and complaining of abdominal pain, the providers could have interpreted her symptoms as mere gastrointestinal discomfort. In this instance, that could have led to an error in diagnosis and serious complications. The receiving ED could have had a cardiac arrest to manage rather than an early, silent acute coronary event. In this instance, contacting medical command, obtaining a focused history and providing general treatment contributed to a positive outcome for the patient. PMID- 12078405 TI - Street drug roulette continues. PMID- 12078407 TI - "We have no choice". EMS in Israel. PMID- 12078408 TI - The Israeli EMS system. PMID- 12078409 TI - Ambulance safety: what's new, what's needed. PMID- 12078410 TI - Lights & sirens. PMID- 12078411 TI - Functional assessment of the elderly patient. PMID- 12078412 TI - Geriatric suicide: helping the hopeless. PMID- 12078414 TI - Higher profile, higher returns? The fundraising arena post-September 11. PMID- 12078413 TI - Seeing your proposal through the reviewer's eyes. PMID- 12078415 TI - Turning customers into clients. PMID- 12078416 TI - [Report of the activities of the National Aerobiological Surveillance Network, 2001]. PMID- 12078417 TI - [Effect of climatic changes on the phenology of plants and the presence of pollen in the air in Switzerland]. AB - The phenologic observations carried out in Switzerland since 1951 show a clear tendency to the earlier appearance of the spring phases of development of the vegetation during last decades. As these phases are strongly influenced by the temperature, these results reinforce the hypothesis according to which the climatic reheating in progress strongly influences the development of the vegetation. The flowering and the release of the pollen of the plants allergens are significant phenologic phases for human health. Modifications in their dates of manifestation lead to a displacement of the periods of allergic risk. The analysis of the aerobiologic data of the Swiss stations over one duration of more than 20 years showed that the species which flower in winter and in spring were subject to the influence of the soft winters of the years 1990 to 2000. In certain areas the pollen of birch, for example, is nowadays waited on average 3 weeks earlier than 20 years ago, whereas the pollen of ash appears with one month in advance. The phenologic and aeropalynologic studies thus prove sensitive witnesses of the climatic change in progress. This last intervenes like an indirect but significant component of human health, in particular when the field of the allergies is considered. PMID- 12078418 TI - [The Poaceae pollen season in France in 2001]. AB - The aim of this study was to determine the main characteristics of the Poaceae pollen season in France (seasonal and daily quantities) in 2001, and to compare them to those of the previous years. The data came from seven volumetric Hirst traps, located in the cities of Amiens, Paris, Lyon, Nimes, Bordeaux, Montlucon and Strasbourg. The study period range from the beginning of March to the end of August. The pollen quantities of 2001 were particularly high in three sites (Montlucon, Bordeaux et Lyon); they were higher than those of 2000 in Amiens and Strasbourg, but in those sites the peak of the whole series was situated in 1994; lastly the pollen quantities of 2001 were little different or lower than those of 2000 in Nimes and Paris. This was probably not without effect on allergic people. PMID- 12078419 TI - [Poaceae pollens. Comparison of Poaceae pollens in 1999 among Marseille, Gap, and Briancon 2000 meters]. AB - Pollens of Poaceae remain the major concern of all those which are concerned with the allergies. The grasses in question are ubiquitaires, but the variations due to climate and the relief influence much on the statements. Our country, from its geographical richness and by the number of sites of collecting, allows an interesting synthesis and a better comprehension of the metabolism of the plant. We are interesting here in the sites of Nimes, Aixen-Provence, Gap and Briancon, included in the National Network. Moreover, one sensor placed temporarily at 2,000 meters of altitude, meadows of Briancon, gives one further information. PMID- 12078420 TI - [Ambrosia pollination: Lyon--Dauphine--Isere, 2001]. AB - Within the framework of the decrees taken on the departments of Isere and Rhone concerning the protection of the population with respect to ambrosia, the object of this study is to relate the analysis of the daily pollen counts in the of air along the Lyon-Grenoble axis. With this intention, three sensors of the type HIRST were positioned according to standards of the European networks in Lyon Gerland, Bourgoin-Jallieu and Grenoble. The results presented show the curves of the daily pollen counts on the three sites and by indicating the allergic thresholds of risks associated defined by the RNSA. The number of days with high risk, for each of the three sites, can thus be compared. Experiment must be prolonged to follow, these comparative results and moreover, possible effectiveness of the obligations of pulling up of the plant on atmospheric pollination. PMID- 12078422 TI - [Ambrosia and the study of its dissemination]. PMID- 12078421 TI - [Ambrosia pollen in Switzerland--produced locally or transported?]. AB - The ambrosia (Ambrosia artemisiifolia L.) has been announced in Switzerland for more than 100 years. During the last decade, signs of an increased presence of the ambrosia and its pollen appear in the south-west of Switzerland, in particular in Geneva. Aerobiologic measurements make it possible to highlight the importance of the transport of pollen by the winds since the area of Lyon, where the ambrosia is abundant. It also bring elements making it possible to detect a local production of pollen in the Geneva area, where several invaded fields were discovered with the autumn 2001. A multidisciplinary group was created in Geneva and it attempts to prevent as far as possible probable invasion of the ambrosia. PMID- 12078424 TI - [Digestive endoscopies: who does what?]. PMID- 12078423 TI - Food allergy and IgE sensitization caused by spices: CICBAA data (based on 589 cases of food allergy). AB - BACKGROUND: Spices originate in various botanical families: Apiaceae, Lamiaceae, Lauraceae, Leguminosae, Liliaceae, Myristicaceae, Myrtaceae, Piperaceae, Solanaceae, Zingiberaceae.... METHODOLOGY: Prick-tests to native spices have been carried out in patients suspected of food allergies to spices. The CICBAA data bank includes 589 cases of food allergies, a part of which has benefited from investigations for spices. Data about the rate of sensitization and food allergy are available. RESULTS: Frequent sensitization to Apiaceae is observed: coriander, caraway, fennel, celery: 32% of prick-tests in children, 23% of prick tests in adults. Sensitization to Liliaceae: garlic, onion, chive, is observed in 4.6% of prick-tests in children, 7.7% of prick-tests in adults. Rare cases of sensitization to paprika and saffron are recorded. Prick-tests to nutmeg, ginger and clove are currently negative. 10 food allergies related to the mugwort-celery spices syndrome are reported: coriander: 1, caraway: 2, fennel: 3, garlic: 3, onion: 1. Food allergy to spices is unfrequent: 2% of the totality of food allergies. However, only adults are allergic to spices and allergy to spices accounts for 6.4% of food allergies in adults. Tiny amount of proteins are usually ingested. Patients at risk of spice allergy are young adults sensitized to mugwort and birch allergens, sharing cross-sensitization with various food vegetal allergens. The clinical suspicion raises from frequent post-prandial systemic reactions. Other allergens of vegetal origin have to be cleared. Diagnosis can be established by DBPCFC using powdered spices in capsules. PMID- 12078425 TI - [Value of cervical plain radiography in unconscious trauma patient]. AB - INTRODUCTION: The aim of this study was to evaluate the diagnostic value of cervical spine standard radiographs, performed in emergency, and compared with entire cervical helical CT with multiplanar reconstructions as reference. STUDY DESIGN: Open prospective study. PATIENTS AND METHODS: We conducted a six months prospective study including all patients over 15 years of age and unconscious (Glasgow Coma Scale < or = 12). Each patient underwent standard radiographs as well as helical CT of the entire cervical spine. Three senior surgeons and one senior radiologist evaluated the standard radiographs quality. The interpretation was performed by 7 different groups of judges. Two radiologists interpreted the helical CT. For each group, the sensibility, the specificity and the count of correct diagnosis for the standard radiographs were evaluated. The results of the correct diagnosis of each group were then compared to determine the most performant group. RESULTS: Fifty-one patients were included. Helical CT diagnosed spine injuries in 11 patients. The quality of standard radiographs was poor with less than 10% judged correct and 90% of the C7-D1 junction not visible. In the best group (radiologist), the sensibility was 50%, the specificity was 85% and the count of correct diagnosis was 78%. For the correct diagnosis, senior radiologist was significantly better than anaesthetist students, radiologist students and emergency physicians. CONCLUSION: The diagnostic value of standard radiographs was weak whatever the physician. Therefore, helical CT of the entire cervical spine is absolutely necessary and must be performed during the initial evaluation, if the haemodynamic conditions are required. PMID- 12078426 TI - [Potential advantages a new syringe pump system]. AB - OBJECTIVES: To evaluate what could be the benefit in terms of alarm suppression, of a new syringe pumps system, which allows automatic relay without alarms. PATIENTS AND METHODS: We have prospectively recorded the nature and the mode of relay of all syringe pumps for administration of drugs to patients in two ICU. The perfusion regimen was studied in 61 patients over the complete duration of their stay in two ICU. Alarms were also recorded in one ICU room, over 13 days consecutive. The records were processed off-line with an automatic detection recognition system in order to assess the origin of each alarm. Accordingly, the amount of alarm corresponding to monitor, ventilator and syringe pumps was computed. RESULTS: We found that syringe pumps, which would provide automatic relays without alarm, would suppress 17% of alarms in ICU and automatize 65% of the relay procedures, which are usually performed often in emergency, on response to the end of perfusion alarm. CONCLUSION: New syringe pumps allow both reduction of alarms in ICU and a better management of nurse workload. Indeed syringe relay can be planned in advance. PMID- 12078427 TI - [Severe malaria in expatriates in an intensive care unit in Abidjan (Ivory Coast)]. AB - OBJECTIVE: To analyse clinical and prognosis aspects of severe malaria in expatriates hospitalized between 1990 and 1999 in the intensive care unit in Abidjan. STUDY DESIGN: Retrospective survey. METHODS: According to the World Health Organization's criteria, the retrospective study of severe cases of malaria who received treatment and care at the intensive care unit. Epidemiological, clinical manifestations and evolution were analysed on each patients. RESULTS: 66 upon 927 expatriates hospitalised in the period of the study, had severe malaria with falciparum Plasmodium (7.12%). The average age was 42 years. Eleven patients took prophylactic treatment (17%). The clinical aspects were neurological (83%) followed by renal failure (48%), haemoglobinuria (48%) and hyperparasitemia (59%). During the hospitalisation we recorded 12 deaths (18%). The criteria that were associated with mortality in pejorative order were: coma (RR = 8.04), respiratory distress (RR = 5.06), metabolic acidosis (RR = 5.06), shock (RR = 3.67) and convulsions (RR = 2.86). CONCLUSION: Severe malaria was frequent and associated with high mortality in expatriates who are living in Africa. This study reinsists the necessity of prophylactic treatment to be reinforced in informing the travellers. This study showed frequency and mortality rate of survey of malaria and the criteria associated with high mortality rate. PMID- 12078428 TI - [Community-acquired peritonitis. Therapeutic follow-up in the case of secondary aggravation: the importance of microbiological results and medical imaging]. PMID- 12078429 TI - [The epidemiology of toxemic infections caused by Staphylococcus aureus in an intensive care setting]. AB - The mission of the National Reference Centre for Staphylococcal Toxemia (CNRTS) is to participate in the epidemiological surveillance of staphylococcal toxemia in France. As these syndromes do not have to be declared, the data collected remain incomplete. Comparison of the clinical data with the results of molecular analysis of the causal strains has nonetheless enabled advances in our understanding of the present-day epidemiology of these syndromes, a clearer knowledge of their pathophysiology and isolation of a hitherto unknown entity, staphylococcal necrotizing pneumonia. PMID- 12078430 TI - [Staphylococci resistant to antibiotic therapy]. AB - Increasing frequency in antibiotic resistance in Staphylococcus aureus is worrying. In this review, we briefly describe the mechanisms of resistance to antibiotics in staphylococci and we report the current incidence of antibiotic resistance in France. Methicillin-resistance is mostly confined to hospitals. However, spread of multiply resistant strains of staphylococci to the community is a putative threat. Strains with diminished susceptibility to glycopeptides are rare but possibly underestimated because of the in vitro difficulties to detect glycopeptide resistance. A continuous resistance survey is necessary. The problem of multiresistance is different for hospital and community staphylococci. The later strains still remain susceptible to many antibiotics, apart from some exceptions. By contrast, incidence of multiply resistance in hospitals remains high. After a recent decrease, incidence of oxacillin resistance seems to be stabilized. A new and recent feature is the great variety of resistance phenotypes which can be found in methicillin-resistant strains, in particular with gentamicin susceptibility. PMID- 12078431 TI - [The importance of detecting methicillin-resistant Staphylococcus aureus in an intensive care setting]. AB - In recent years, the fight against nosocomial infections has constituted a major public health challenge. Measures aimed at preventing the spread of multiresistant bacteria (MRB) through carriage combine the more or less systematic detection of carriers and precautions concerning isolation. In the light of published data, elements for the evaluation of a cost-efficiency ratio for measures concerning detection and isolation in intensive care are presented. The high levels of methicillin-resistant Staphylococcus aureus (MRSA) observed in hospitals constitute the principal target. Although isolation precautions appear to be effective, the value of detecting MRSA remains a subject of debate. Such detection has several objectives: the identification of carriers, assistance in the implementation of isolation precautions, the eradication of carriage and the evaluation of strategy. These different objectives are discussed. PMID- 12078432 TI - [Therapeutic principles of staphylococcal infections--Role and limitations of standard compounds]. AB - Antibiotic therapy plays an important (but not exclusive) role in the treatment of staphylococcal infections. Measures aimed at reducing the bacterial inoculum through local procedures must be envisaged as often as possible. The removal of any foreign, infected materials is essential to success. In this article, we review the different, active antibiotics available, their advantages and disadvantages and their indications. In the light of these data, we propose a therapeutic approach to severe bacterial infection caused by a cluster of Gram positive cocci. Staphylococcal infections pose daily therapeutic problems, whether in open-care practice or intensive care units. The specificity of staphylococcal infections encountered in an intensive care setting require a therapeutic approach which takes account of the context, and particularly of the incidence of resistant staphylococcal infections. PMID- 12078433 TI - [Antibiotic combinations in Staphylococcus aureus infections: arguments against]. AB - In a case of severe infection due to Staphylococcus aureus, generally accepted practice involves the use of a combination of antibiotics. This article examines the type and quality of the experimental and clinical data which has been collected in this context. Published findings suggest that the arguments in favour of combinations are usually theoretical, and that a demonstration of the superiority of combinations has never been made by comparative, randomised studies. PMID- 12078434 TI - [Antibiotic combinations in Staphylococcus aureus infections: arguments in favour]. AB - More than 40% of Staphylococcus aureus strains isolated in hospitals are methicillin-resistant, thus posing daily problems regarding therapy. The treatment of these infections is principally based on glycopeptides. The need or not to combine them, first-line or second-line, with another antibiotic, remains a subject for debate. In this article, we have endeavoured to demonstrate the validity of arguments which plead in favour of using glycopeptides in combination, and not as single-drug therapy. PMID- 12078435 TI - [Profile of patients facing therapeutic problems in nosocomial infections due to methicillin-resistant staphylococci--A survey of practices]. AB - Methicillin-resistant staphylococcal infections (MRSI) are still common in an intensive care setting. Their management is mainly based on glycopeptides, combined with other antibiotics when this is possible, and also on treatment of the portal of entry (removal of foreign bodies, surgery...). Implementation of this antibiotic therapy may meet with difficulties linked to the micro-organism (existence of strains with diminished sensitivity to glycopeptides), to the toxicity of glycopeptides or to the unfavourable course of the infection. A survey of practices was performed on a representative sample of 240 intensive care units in France. Glycopeptides, and particularly vancomycin, were the most frequently employed and prescribed in combination with other compounds. Therapeutic problems were considered as occasional, their incidence ranging from 0 to 50%. The problems reported were mainly linked to adverse effects: most frequently renal toxicity and, to a lesser extent, immunological and allergic complications. Diminished sensitivity to glycopeptides was only reported by a third of physicians, and this sporadically. Such a survey of practices is an essential preliminary to an epidemiological study of the incidence of MRSI and related therapeutic problems. PMID- 12078436 TI - [The use of glycopeptides in intensive care and anaesthesia]. AB - With the objective of clarifying the modes of using glycopeptides in intensive care, a survey with a declared intention was performed in June 2001, in the form of a postal questionnaire; it was possible to exploit 742 answers. Analysis of the results showed that 15% of the doctors completing the questionnaire had not employed glycopeptides within the past six months. Preference was given to vancomycin, and 65% of practitioners prescribed this drug only, while 1% of them only prescribed teicoplanin. For vancomycin, a central route is used in 9 out of 10 cases, with a preference for continuous infusion (69% versus 48%). A loading dose is prescribed in 70% of continuous infusions (> or = 15 mg.kg-1 in 69% of cases), and in 19% of intermittent infusions (> or = 15 mg.kg-1 in 90% of cases). The mean vancomycin dosage is 30 mg.kg-1.d-1; in the case of intermittent administration this involves two (51%), three (17%) or four (27%) injections per day. The target concentrations are residual serum vancomycin levels of between 13.5 and 23.6 mg.L-1 with intermittent administration or plateau levels of 18.3 to 29.4 mg.L-1 with continuous infusion. Teicoplanin is administered intravenously (92%) and/or via the i.m. route (34%). The mean dosage is < 10 mg.kg-1.d-1 in seven out of ten cases. A loading dose is administered every 12 hours at the same dosage in 77% of cases (< or = 3 injections for 65% of prescribers). The residual teicoplanin concentrations sought are between 14.3 and 25 mg.L-1. Monitoring of serum levels is ensured at least once a week in 97% of cases with vancomycin and 66% of cases with teicoplanin. In conclusion, is seems that the methods of using glycopeptides vary considerably. The great heterogeneity of practices suggests a lack of compliance by prescribing practitioners with current recommendations. It also seems that a precise definition of the target plasma levels to be achieved in different indications is necessary. PMID- 12078437 TI - [The management of treatment failures for staphylococcal infections]. AB - Therapeutic failure is a situation to be feared in severe infections due to staphylococci. In this article, we consider the difficulties faced is achieving a positive diagnosis in the context of such failures, and an approach to analysing their causes. The possible reasons for therapeutic failure must be considered systematically. The two principle causes are unquestionably the persistence in the patient of an infected, foreign body (intravenous catheter or prosthesis) and endovascular infections (thrombophlebitis or endocarditis). The rules for the prescription of antibiotics are followed, target concentrations are achieved and drug failures are a rarity today. This observation may be called into question by the emergence of strains with diminished sensitivity to glycopeptides. These therapeutic failures remain exceptional cases today. PMID- 12078438 TI - [Novel compounds active on staphylococci]. AB - Research efforts to discover new compounds active against staphylococci are more than ever justified today. The incidence of methicillin-resistant staphylococci remains very high in hospitals, and the solution provided by glycopeptides is far from being satisfactory. These compounds exhibit mediocre pharmacokinetic and pharmacodynamic properties. Their ease and safety of use are poor. Finally, strains with diminished sensitivity to these antibiotics are beginning to appear. This article examines the opportunities offered by two new anti-staphylococcal agents: quinupristine-dalfopristine (Synercid) and linezolide (not marketed in France). PMID- 12078439 TI - [Amniotic fluid embolism: successful evolution course after uterine arteries embolization]. AB - A 28-year-old woman, G3P3, who was otherwise healthy and had taken no medication and had no known allergy, was admitted to our hospital for delivery after a normal pregnancy. An epidural catheter was inserted for analgesia and labour was induced with oxytocin. Two hours later, she suffered a sudden cardiac arrest. She was immediately treated and, since a normal cardiac rhythm and a blood pressure of 90 mmHg has been obtained 30 minutes later, a 3750 g child was delivered by caesarean section. Soon after delivery, a life-threatening uterine haemorrhage appeared, due to DIC. Evolution was favourable, after bilateral uterine arteries embolization had been performed. The diagnosis of amniotic fluid embolism was established by the clinical course, the absence of local cause and the presence of a large number of amniotic cells in the mother's peripheral blood. Tryptase blood concentration was normal in the mother's blood. PMID- 12078440 TI - [Hemoperitoneum and pregnancy in a patient with von Willebrand's disease type 3]. AB - A 24-year-old woman with type 3 (characterised by a blood level of von Willebrand factor less than 1%) was admitted to the surgical intensive care unit with an haemorrhagic shock. She was pregnant (Beta HCG = 115 Ul.L-1), the echography of the abdomen, soon after admission, revealed a haemoperitoeum. She underwent abdominal surgery after substitution treatment. The surgeon found an ovarian haemorrhagic cyst. Later the course of pregnancy was normal and she was delivered by caesarean section. We describe the therapeutic strategy of that care. PMID- 12078444 TI - [Migration of desilet during pose of central venous way]. PMID- 12078443 TI - [Hypnosis as treatment for nausea and vomiting: it works!]. PMID- 12078441 TI - [Coronary thrombosis on patient with the factor V Leiden mutation]. AB - We report a documented observation of coronary thrombosis occurring in a 25-year old patient with no risk factor, presenting a hereditary thrombophilia (facteur V Leiden) diagnosed a few months earlier in a context of venous thrombosis. This patient had a spread out anterior myocardial infarction with cardiac arrest due to a ventricular fibrillation; although he was quickly rescued by the mobile intensive Care Unit, the patient died 48 hours later, after cerebral anoxia. The mutation called factor V Leiden is a widely spread hereditary family thrombophilia (5 to 6% of the population) and is characterized by a resistance to activated C protein provoking a hypercoagulable state. The unexpected arterial thrombosis, very rare in that case, can be extremely serious and raises the question of a preventive medication such as antiplatelet agent or low-molecular weight heparin as soon as the genetic abnormally has been proved to be symptomatic. PMID- 12078445 TI - [Weakness during diving: search for monoxide poisoning]. PMID- 12078442 TI - [Fusarium pleural effusion after a ventricular assist device]. AB - We report the case of a 36-year-old man with a pleural effusion that complicates the postoperative period after the implantation of a ventricular assist device (VAD). The epidemiological, etiologic and therapeutic features of Fusarium infections were reviewed. Complete recovery of the infection was obtained after a treatment by liposomal amphotericine B (AmBisome) and 5 fluorocytosine. PMID- 12078446 TI - [Laryngeal mask for adenoidectomy: some useful cautions]. PMID- 12078447 TI - [Guidelines for prehospital care of patients in grave condition]. PMID- 12078448 TI - Communication links between service providers is essential. PMID- 12078449 TI - Use of laryngeal mask airway in pre-hospital environments. PMID- 12078450 TI - Developing a nursing observation chart. PMID- 12078451 TI - Hair dye and henna tattoo exposure. PMID- 12078452 TI - Improving the efficiency of patient handover. PMID- 12078453 TI - Common cardiac arrhythmias. AB - The interpretation of cardiac arrhythmias plays an important role in the diagnosis of cardiac disorders. Helen Hand examines the physiology of the heart, the electrical conduction mechanism, cardiac rhythm disorders and cardiac monitoring. PMID- 12078454 TI - [Hypertensive inferior esophageal sphincter. Prevalence and clinico-manometric features]. AB - BACKGROUND: The hypertensive lower esophageal sphincter (HLES) is an unusual primary motor disorder of the esophagus. The lack of a precise definition has made it difficult to establish its prevalence and its clinical and manometric characteristics. OBJECTIVES: 1) To determine the prevalence of HLES using precise diagnostic criteria, 2) To identify the clinical manifestations of this disease, and 2) To compare manometric findings in patients with HLES against normal subjects. METHODS: A total of 705 consecutive esophageal manometries were performed over 52 months. HLES was defined as basal pressure 45 > or = mmHg, normal relaxation in response to swallowing, and normal peristalsis of the esophageal body. Clinical records were reviewed to identify the main symptoms that prompted the manometry. The manometric data of patients with HLES was compared to that of 16 healthy control subjects. RESULTS: Seven HLES patients were identified establishing a prevalence of 1%. Four HLES cases were referred due to dysphagia, two due to symptoms of gastroesophageal reflux disease, and one due to thoracic pain. Upon comparing manometric data of patients with HLES and healthy control subjects the motor function of the esophageal body, the total length of the lower esophageal sphincter, and its position in respect to the diaphragm were similar in both groups. CONCLUSIONS: Prevalence of HLES was 1%. Dysphagia was the main symptom. Total length of the sphincter, as well as length of the thoracic and abdominal segments do not appear to contribute to high basal pressure. PMID- 12078455 TI - [Evaluation of 20 years of experience and quality of life in patients surgically treated for liver cystic disease]. AB - Surgical treatment of liver cystic disease is reserved for symptomatic patients. The surgical approach is chosen according to the size and distribution of the cysts. In patients with massive hepatomegaly secondary to polycystic liver disease, liver transplantation is indicated with excellent results and quality of life. OBJECTIVE: To evaluate over 20-year period, the results in terms of clinical outcome of three groups of patients with cystic liver disease (EQ) who received surgical treatment and to determine postoperative quality of life. MATERIALS AND METHODS: In a 20-year period, 44 patients were operated on; 24 had simple liver cyst, 13 had polycystic liver disease, and seven cystadenomas. Using the SF36 questionnaire, self-perception of quality of life was evaluated using eight scales in two major categories: Physical component summary (PCS) and mental component summary (MCS). RESULTS: Upper abdominal pain was the main clinical symptom. Fenestration was the most frequent procedure performed. No differences in quality of life were observed in all, while good quality of life was recorded in all groups at a median follow-up of 39 months. CONCLUSIONS: Surgical treatment of cystic liver disease is reserved for symptomatic patients or complications such as rupture, infection, and hemorrhage. Therapeutic alternatives should been chosen on an individual basis. Good quality of life is obtained after surgery in these patients. PMID- 12078456 TI - [Surgical treatment of colorectal cancer. 10-year experience at the INCMNSZ]. AB - OBJECTIVE: To analyze the short- and long-term outcomes of the surgical treatment for colorectal cancer at the Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran (INCMNSZ) in Mexico City. BACKGROUND: Colorectal cancer is the second most frequent malignant tumor of the digestive tract in Mexico. Its incidence is rising, and its identification in early stages is important to offer a curative surgical treatment. METHODS: Retrospective, descriptive observational trial. We studied 165 patients who underwent colorectal surgery for treatment of cancer between 1985 and 1994 at the INCMNSZ. We compared the variables of patients and the surgical procedures and their relationship with morbidity and mortality with 5-year survival. RESULTS: Survival was greater in curative attempting procedures of the colon than of the rectum (p < 0.05). Survival rate was also greater in earlier stages of the disease at the moment of diagnosis. Surgical morbidity occurred in 30% due to wound infections in most cases. Mortality rate was 3.6%, while the main cause was sepsis. Patients > 65 years of age had a higher mortality rate. Rectal surgery had higher morbidity than colonic surgery. CONCLUSIONS: Detecting patients at earlier stages of the disease is required to offer them a curative-attempting surgery procedure. Stage at moment of diagnosis correlates with survival rates. Age was a risk factor for surgical mortality, and rectal surgery, a risk factor for morbidity. PMID- 12078458 TI - [Hamartoma of the bile duct. Report of a case]. AB - BACKGROUND: Gallstones and neoplasms generally cause obstructions in the biliary duct. There are some infrequent problems that may lead to obstructions, such as hamartomas of the biliary duct. CASE REPORT: A 34-year-old diabetic female patient begins with abdominal pain and jaundice. The diagnostic approach initiated with hepatobiliary ultrasound that reported cholelithiasis and enlargement of the biliary ducts. Cholangiopancreatography was subsequently performed and a biliary tree lesion was discovered; the patient underwent cholecystectomy with exploration of the biliary ducts. Clinical evolution was satisfactory. Pathologic findings indicated hamartoma of the biliary duct. PMID- 12078457 TI - [Etiology of intestinal occlusion]. AB - BACKGROUND: Intestinal obstruction is one of the leading causes of admission to emergency wards around the world, and its etiology has changed over the past century. AIM: The goal of this study was to ascertain the causes of intestinal obstruction at our Institution and compare the results with other reports of Mexican, U.S., and Canadian hospitals. METHODS: Retrospective review of a cohort of patients with intestinal obstruction operated on between 1985 and 2000. Demographic data and operative findings were obtained. RESULTS: Our cohort included 452 patients, 55.3% were women; mean age for the entire group was 54 years. The obstruction was located in the small bowel in 86.9% of cases and the leading causes were adhesions (58.6%), hernia (16.1%) and neoplasia (13.9%). Other etiologic factors had a low incidence that varied between 4.4 and 0.22%. CONCLUSIONS: The most common causes of intestinal obstruction are similar to those reported in the U.S., British and Canadian medical literature. Some tertiary-level Mexican institutions showed the same incidence of etiology, but some large general hospitals in Mexico City showed etiologic factors reported 100 years ago by the current so-called developed countries, i.e., that the same socioeconomic conditions existed in both population groups nearly a century apart. PMID- 12078459 TI - [Retrograde intussusception posterior to gastrojejunal anastomosis. Report of a case]. AB - OBJECTIVE: We report a case of retrograde jejunogastric intussusception in a patient with previous gastric surgery who required surgical treatment for its resolution. BACKGROUND: While intussusception is relatively common in children, it is infrequently seen in adults. Five percent of all cases occur in adults and in 90% of these, there is a lead point, a well-definable pathologic abnormality. Retrograde jejunogastric intussusception is a rare complication after gastrojejunostomy. The most common symptoms are pain, nausea, vomiting and blood per rectum. Diagnosis of adult intussusception is often difficult and is base in clinical suspicion and noninvasive techniques. METHOD: A case of a retrograde intussusception in a patient after gastric surgery who presented with abdominal pain and gastrointestinal bleeding. RESULTS: The patient underwent surgical treatment with reduction of the intussusception. The postoperative evolution was satisfactory. CONCLUSIONS: Management of intussuception in adults is usually laparotomy and in 90% of cases, a pathologic lesion will be identified. Retrograde jejunogastric intussusception is also managed surgically and simple reduction is the accepted form of treatment in most cases. PMID- 12078460 TI - [Esophageal perforation caused by balloon dilatations in a patient with achalasia]. AB - OBJECTIVE: To present a case of esophagic perforation due to a pneumatic dilatations of the cardias in a patient with achalasia. BACKGROUND: Achalasia of the esophagus is a primary motor alteration with an unknown cause. The principal characteristics are the lower or absent peristaltic waves in the esophagic body and the impared lower esophagic sphincter relaxation during deglution. The management is palliative with pneumatic dilatations, pharmacologic therapy or with surgery. When a esophagic perforation occurs, it must be diagnosed and treated as soon as possible because a delay in diagnosis and management affect directly the patient prognosis. METHOD: To present a case of esophagic perforation due to pneumatic dilatations of the cardias in a patient with achalasia. CONCLUSIONS: An useful palliative management for achalasia is the pneumatic forceful dilatation, however it carries some risk of perforation; in every patient who was treated with dilatations is mandatory to think in perforation when the clinical condition is different after the procedure. The success of surgical management of the perforated esophagus and the control of sepsis and mediastinitis depends directly of the clinical condition of the patient and the time within perforation and surgical therapy. PMID- 12078461 TI - [Historical features of surgical anatomy of bile ducts and cholecystectomy]. PMID- 12078462 TI - A follow-up survey of the use of complementary and alternative medicines by surgical patients. PMID- 12078463 TI - Nurse anesthesia students and liability. PMID- 12078464 TI - Monitoring nurse anesthesia educational programs. PMID- 12078465 TI - CRNA-conducted research: is it being done? AB - An emphasis on research was established with the 1998 Council on Accreditation of Nurse Anesthesia Educational Programs (COA) requirement that all nurse anesthetists graduate with a master's level of education. Nursing research is essential to expand the profession's knowledge base and to establish a foundation for evidence-based practice. However, to date there are no existing data indicating whether practicing CRNAs are conducting research or to what extent. The purpose of this study was to determine whether research is conducted by Certified Registered Nurse Anesthetists (CRNAs) and what factors influence whether CRNAs engage in research. A quantitative descriptive design was used. Random sampling from the total AANA membership roster was done. A questionnaire developed by the researchers and evaluated by content experts was used to obtain data after the questionnaire was validated with a pilot study. Of 370 questionnaires mailed, 173 (46.8%) were returned. Results revealed a small percentage of CRNAs are involved in the research process at some level. The majority of CRNAs conducting research did not receive preparation in the research process from their educational curriculum. There was a significant relationship between working in a teaching hospital and whether CRNAs conducted research. Many barriers were identified. PMID- 12078466 TI - Continuous spinal anesthesia for cesarean section for a morbidly obese parturient patient: a case report. AB - This case report details the successful anesthetic management of a morbidly obese parturient patient who presented for a repeat, elective cesarean section. The preanesthetic evaluation and the indications for choosing a continuous spinal anesthetic are discussed. Evaluation of the anesthetic plan is also discussed through a review of the postoperative scenario. PMID- 12078467 TI - A synthesis of the Australian Patient Safety Foundation Anesthesia Incident Monitoring Study, the American Society of Anesthesiologists Closed Claims Project, and the American Association of Nurse Anesthetists Closed Claims Study. AB - The study of anesthesia risk is fraught with methodological challenges and the epidemiological uncertainty peculiar to anesthesia: the true frequency of anesthetic mortality is unknown. If anesthesia mortality is as rare as 1 in 200,000 cases, the sample needed to study this phenomenon would be enormous. Existing studies provide insights to the genesis of damaging events and adverse outcomes in anesthesia. We examined research design, methodology, and findings to date in 3 studies of anesthesia risk. Limitations include self-report by providers in the Australian study and the retrospective nature of closed claims research in American studies. Respiratory events were the largest class of injury in all 3 studies; substandard care frequently was involved. Australian investigators noted a high rate of human error and equipment issues. American physician investigators found that death or brain damage occurred in 85% of respiratory cases, 72% of which were deemed preventable. Nurse anesthetist investigators had similar findings for respiratory claims. Patient acuity and procedure complexity may be less significant contributory factors to anesthesia risk than are provider vigilance and clinical decision making. Prospective multicenter studies conducted politically may be the only type of research that definitively addresses the myriad issues in anesthesia risk research. PMID- 12078468 TI - Is there postoperative evidence of implicit learning following aural stimuli at moderate hypnotic BIS levels during general anesthesia? AB - Patients may develop behavioral changes after the administration of a general anesthetic without a triggering stimulus. This phenomenon, referred to as implicit learning, continues to be debated. Some researchers have denied the phenomenon takes place, while others have demonstrated behavioral changes not related to recall. In previous studies it was difficult to determine the effects of taped messages or suggestions, as anesthesia techniques were not standardized, and true depth of anesthesia was impossible to determine. The development of bispectral index (BIS) monitoring allows practitioners to determine the depth of anesthesia based on neuronal activity. In the present study, we compared 2 randomized groups of patients from a convenience sample of patients scheduled to undergo elective spinal surgery at a Department of Veterans Affairs Medical Center. All patients were screened for normal hearing acuity. The control group listened to tapes of white noise, while the experimental group listened to a word list and short story selection. Both groups listened to the tapes at similar BIS levels. A posttest was administered to determine explicit recall, as well as implicit memory formation. No patients had explicit recall. There was no difference in the implicit memory scores between patients in the experimental group and the control group. PMID- 12078469 TI - Acute fatty liver of pregnancy: a case report. AB - The third trimester of pregnancy has several unique disease entities that challenge the anesthetist. Acute fatty liver of pregnancy (AFLP) is an increasingly recognized metabolic disorder found late in pregnancy. Current research estimates the incidence of AFLP at 1 per 6,659 births. Improved maternal morbidity and mortality is credited to early recognition and termination of the pregnancy. The decline in hepatic cellular activity is evident by the deterioration in metabolic, synthetic, and excretory functions of the liver. This obstetrical emergency can lead to death of both mother and child if not diagnosed in time to prevent coagulopathic complications. PMID- 12078470 TI - Nurse anesthesia students' perceptions of the anesthesia patient simulator: a qualitative study. AB - The purpose of this study was to describe the perceptions of nurse anesthesia students (NAS) who used a MedSim simulator (MedSim USA, Inc, Ft Lauderdale, Fla) as part of their educational training. A convenience sample of 12 NAS in their first year of clinical training was researched. The researcher analyzed data qualitatively from observations made during 4 different sessions. Session 1 introduced the students to the simulator. Session 2 involved each student performing an anesthetic induction. A minor incident such as hypotension, hypertension, bradycardia or tachycardia occurred in session 3, and a major incident such as cardiac ischemia, anaphylaxis, bronchospasm, or malignant hyperthermia occurred in session 4. Data collection involved observation by the primary investigator, journal entries by the anesthesia students, and focus group interviews with the students. Results of the study indicate that, although students experience feelings of apprehension, uneasiness, or fear during the sessions, they felt it was very educational. Disadvantages include the lack of reality, lack of knowledge on handling crisis events, possibility of fixation errors, and the presence of anxiety. Advantages include improved critical thinking and decision-making skills, increased confidence, and improved clinical preparation. Results can be used to assist instructors in improving the students' learning experiences and to teach more effectively. PMID- 12078471 TI - AANA journal course. Update for nurse anesthetists. A common sense approach to hypothermia. AB - Hypothermia frequently is considered inadvertent in the perioperative setting. The preservation of vital body heat has been an issue since the 1800s. This article provides a select review of the causes of hypothermia and the methods for prevention of hypothermia during the perioperative period. Providing patients with an environment designed to foster normothermia can preclude the costs of longer hospital stays, prevent morbid conditions associated with hypothermia, and provide patients with a more comfortable perioperative experience. Our goal as perioperative healthcare providers is a normothermic perioperative experience for all patients. PMID- 12078472 TI - Nitric oxide production by Leishmania-infected macrophages and modulation by cytokines and prostaglandins. AB - Nitric oxide (NO) produced by an inducible nitric oxide synthase (iNOS or NOS2) plays a major microbicidal role in murine macrophages and its importance is now emerging also in the dog and human models. In dogs we demonstrated that macrophages in vitro infected with Leishmania infantum produced NO, after stimulation with cytokine-enriched peripheral blood mononuclear cell supernatants. In addition, parasite killing was reduced by the NOS inhibitor L-NG monomethylarginine. On the contrary, canine blood monocytes before macrophage differentiation did not release NO, and their leishmanicidal activity was instead correlated with superoxide anion and interferon (IFN)-gamma production. In human macrophage cultures, after infection with Leishmania infantum, we showed both iNOS expression by immunofluorescence and western blotting and NO release by the Griess reaction for nitrites. Various cytokines and prostaglandins can differently modulate NO synthesis. In our experiments, stimulation by recombinant human IFN-gamma and bacterial lipopolysaccharide greatly enhanced iNOS expression and NO production in human macrophages. In addition, the prostaglandin E2 increased NO release in activated, Leishmania-infected human macrophages. These results are interesting in the light of a possible immunological or pharmacological regulation of NO synthesis and microbicidal functions of macrophages. PMID- 12078473 TI - Study on immunobiology in endoparasites of public health interest: echinococcosis hydatidosis. AB - Cystic echinococcosis caused by Echinococcus granulosus is one of the most widespread zoonoses in the Mediterranean, endemic in some regions such as Sardinia. Some aspects of the research conducted in this area are briefly reviewed for an integrated analysis of both epidemiological and immunobiological knowledge, gained from field observations and experimental studies. Data on epidemiology in intermediate hosts, immunological assessment of exposure in humans, immune response and Th1/Th2 polarization in secondary experimental hydatidosis, kinetics of response in definitive host and cytokine production in experimental models are briefly reported. They confirm the usefulness of an immunobiological approach both in intermediate and definitive hosts and their potential in prevention, monitoring and control of E/H. PMID- 12078474 TI - Allozyme and PCR-RFLP markers in anisakid nematodes, aethiological agents of human anisakidosis. AB - In the framework of the researches granted by MURST COFIN97, studies on anisakid nematodes, aethiological agents of human anisakidosis, were carried out. The project was aimed to implement the knowledge on the systematics, genetics, ecology and epizootiology of species of the genera Anisakis and Pseudoterranova by applying genetic markers obtained from multilocus allozyme electrophoresis and from PCR-based techniques. The results obtained by allozyme studies allowed to extend the geographic distribution and to detect new definitive and intermediate/paratenic hosts of two sibling species of the A. simplex complex, i.e. A. simplex s.s. and A. pegreffii and to characterise the species A. simplex C, a new sibling species within the A. simplex complex as well as a new species belonging to the genus Anisakis, A. ziphidarum. Combined allozyme and morphological analyses provided markers for the identification of the four sibling species of the Pseudoterranova decipiens complex and their nomenclatural designations. New markers based on PCR-RFLP analysis were used for the identification of sibling species of the Anisakis simplex complex and of another four species of the genus. Genetic markers based on three diagnostic restriction enzymes allowed the identification as A. pegreffii of a larva obtained by endoscopy in a case of human anisakidosis in Southern Italy. PMID- 12078475 TI - Study on immunobiology in ectoparasites of public health interest: Rhipicephalus sanguineus. AB - The recently defined range of Mediterranean Spotted Fever (MSF) in Sardinia coincides with that of the widely studied Rickettsia conorii vector, the brown dog tick Rhipicephalus sanguineus. This tick is major vector of disease in humans, domestic and wild animals. Characteristic features of the vector with respect to their seasonal activity, abundance as well as incidence of human-tick contact and how these factors influence the incidence of MSF were studied. This paper also looks at host, pathogen and tick vector interactions, re-examining some of the immunobiological parameters involved. However, the primary objective of this research is to develop a strategy that will allow better and safer tick control. PMID- 12078476 TI - [Pathogenic protozoans in man: differential characterization of Entamoeba histolytica/Entamoeba dispar complex, Acanthamoeba spp., Microsporidia]. AB - The review summarizes the results in the main parasitological topics of our Lab: amoebic infections due to Entamoeba histolytica/Entamoeba dispar complex and to Acanthamoeba spp. respectively, and human infections caused by microsporidia. Different rapid and advanced techniques have been included in the standardized diagnostic protocols for each topic, and a critical comparison among them was made, in order to define the gold standard diagnostic method: a) E. histolytica/E. dispar: in vitro culture, zymodeme typization, biomolecular identification (PCR), immunoenzymatic assay (ELISA) for direct detection in stools of specific surface antigenic lectins; b) Acanthamoeba spp.: in vitro culture, light and ultrastructural characterization, species identification by immunofluorescence method with monoclonal antibodies, in vitro pharmacological studies; c) Microsporidia: ultrastructural (TEM), biomolecular (PCR), biochemical and immunological (SDS-PAGE, Immunoblotting) studies for species identification, use of advanced ultrastructural techniques ("freeze-etching", "deep-etching") in order to deepen the spore wall structure, to study the cytoskeletal function of actin and to define the mode of infection, in vitro pharmacological assays on some inhibitors of chitin-synthases. PMID- 12078477 TI - Ultrastructural characteristics, biological activity and pharmacological relevance of synthetic malaria pigment (beta-haematin). AB - Malaria pigment (haemozoin, HZ) is the detoxification product of haemoglobin derived haem of intraerythrocytic malaria parasites. At schizont rupture, haemozoin accumulates inside host phagocytic cells. The chemical structure and the spectroscopic characteristics of haemozoin are identical to those of beta haematin (BH), a synthetic pigment obtained from Ferriprotoporphyrin IX (Fe (III) PPIX) in acidic conditions. The process of BH formation is the target of quinoline antimalarials. Here, we summarise the results of our studies on the ultrastructural characteristics, biological and pharmacological relevance of synthetic vs. native haemozoin. 1) By electron microscopy, native HZ and synthetic BH appear as dark brown crystals, morphologically indistinguishable and are internalised by phagocytes at the same extent. 2) Both HZ and BH modulate the production of cytokines (TNF and NO) and increase the susceptibility to lipid peroxidation of mouse or human phagocytes. The antioxidant status of the phagocytes regulates the susceptibility to BH/HZ-mediated effects. 3) The process of BH formation from Fe(III)PPIX, hence haem detoxification, can be inhibited by electrochemically-reduced, Fe(II)PPIX molecules. Maintaining iron in the reduced state can thus be considered a new pharmacological target. This was confirmed by the observation that thiol-reducing agents (NAC, cystein) were able to inhibit BH formation and were toxic to parasites in vitro. PMID- 12078479 TI - Protein degradation and the generation of MHC class I-presented peptides. AB - Over the past decade there has been considerable progress in understanding how MHC class I-presented peptides are generated. The emerging theme is that the immune system has not evolved its own specialized proteolytic mechanisms but instead utilizes the phylogenetically ancient catabolic pathways that continually turnover proteins in all cells. Three distinct proteolytic steps have now been defined in MHC class I antigen presentation. The first step is the degradation of proteins by the ubiquitin-proteasome pathway into oligopeptides that either are of the correct size for presentation or are extended on their amino-termini. In the second step, aminopeptidases trim N-extended precursors into peptides of the correct length to be presented on class I molecules. The third step involves the destruction of peptides by endo- and exopeptidases, which limits antigen presentation, but is important for preventing the accumulation of peptides and recycling them back to amino acids for protein synthesis or production of energy. The immune system has evolved several components that modify the activity of these ancient pathways in ways that enhance the generation of class I-presented peptides. These include catalytically active subunits of the proteasome, the PA28 proteasome activator, and leucine aminopeptidase, all of which are upregulated by interferon-gamma. In addition to these pathways that operate in all cells, dendritic cells and macrophages can also generate class I-presented peptides from proteins internalized from the extracellular fluids by degrading them in endocytic compartments or transferring them to the cyotosol for degradation by proteasomes. PMID- 12078478 TI - Insecticide treated curtains and allelic polymorphism of Plasmodium falciparum genes in a rural area of Burkina Faso (west Africa). AB - To assess the possible impact of insecticide treated curtains (ITC) on the composition of a Plasmodium falciparum population in a rural area of Burkina Faso, blood samples were collected during the rainy season of 1997 from 226 children aged 3-6 years, from 4 villages equipped with ITC and 2 control villages without ITC. The analysis of fragment lengths of 3 highly polymorphic P. falciparum genes (msp-1, msp-2 and glurp) revealed a maximum number of 3 alleles per infected person for each gene. The mean number of clones per infected person was similar in villages with and without ITC. PMID- 12078480 TI - Cytokine memory of T helper lymphocytes. AB - Memory is one of the key features of the adaptive immune system. Specific T and B lymphocytes are primed for a particular antigen and upon challenge with it will react faster than naive lymphocytes. They also memorize the expression of key effector molecules, in particular cytokines, which determine the type and scale of an immune reaction. While in primary activations differential expression of cytokine genes is dependent on antigen-receptor signaling and differentiation signals, in later activations the expression is triggered by antigen-receptor signaling and dependent on the cytokine memory. The molecular basis of the cytokine memory implies differential expression of transcription factors and epigenetic modifications of cytokine genes and gene loci. GATA-3 for Th2 and T bet for Th1 cells expressing interleukin-4 or interferon-gamma, respectively, are prime candidates for key transcription factors of cytokine memory. The essential role of epigenetic modifications is suggested by the requirement of DNA synthesis for the establishment of a cytokine memory in Th lymphocytes. At present the molecular link between transcription factors and epigenetic modifications of cytokine genes in the establishment and maintenance of cytokine memory is not clear. The initial cytokine memory is not stable against adverse differentiation signals, while in repeatedly stimulated lymphocytes it is stabilized by a variety of mechanisms. PMID- 12078481 TI - Ig gene hypermutation: a mechanism is due. PMID- 12078482 TI - Generalization of single immunological experiences by idiotypically mediated clonal connections. AB - Clonal interactions of B cells by idiotope-specific mutual recognition of their antigen receptors with the participation of T cells were assumed to form a web of unknown density, referred to as the idiotypic network. Although these clonal connections were proposed to fulfill important internal regulatory functions, their biological significance, especially in relation to antigen-induced immune responses, remained a mystery. In view of this, we postulate that the basic function of the idiotypic internal connection between B and T cell antigen receptors is to transform antigen-induced cellular activations, by idiotypic crossreactivity, into the regulation of cell clones with different antigen specificities. This process leads not only to the suppression of major clones but also to the activation of minor ones. The latter activating property may allow the generalization of single antigenic experiences, so that the immune system in its entirety benefits in its battle against environmental microbes. Such idiotypic clonal interactions are particularly effective in early ontogeny. During a short neonatal imprinting period, maternal immunological knowledge in the form of somatically mutated, high-affinity IgG antibodies, acquired through a continuous encounter with external antigens, guides the initial ontogenetic development of the immune system and so exerts long-lasting transgenerational advantageous effects in the offspring. PMID- 12078483 TI - The aging of the immune system. PMID- 12078484 TI - Proteolysis and antigen presentation by MHC class II molecules. PMID- 12078485 TI - Predicting redox state of cysteines in proteins. PMID- 12078486 TI - Detection and affinity purification of oxidant-sensitive proteins using biotinylated glutathione ethyl ester. PMID- 12078487 TI - Redox role for tetrahydrobiopterin in nitric oxide synthase catalysis: low temperature optical absorption spectral detection. PMID- 12078488 TI - Lysozyme-osmotic shock methods for localization of periplasmic redox proteins in bacteria. PMID- 12078489 TI - Alterations in membrane cholesterol that affect structure and function of caveolae. AB - Most of the available methods for modifying caveolae structure and function depend on altering the cholesterol content of caveolae. The most important aspect of each method is to ensure the reagents are working in the cells that are being studied. The idiosyncrasies of each method are such that they cannot be universally applied without carefully optimizing the conditions. When used correctly, these methods are accepted as a specific way to perturb the structure and function of caveolae. PMID- 12078490 TI - Contribution of neelaredoxin to oxygen tolerance by Treponema pallidum. PMID- 12078491 TI - Redox control of integrin adhesion receptors. PMID- 12078493 TI - Redox properties of vanillyl-alcohol oxidase. PMID- 12078492 TI - Heme oxygenase 1 in regulation of inflammation and oxidative damage. PMID- 12078494 TI - Anaerobic oxidations of myoglobin and hemoglobin by spectroelectrochemistry. PMID- 12078495 TI - Cysteine-nitric oxide interaction and olfactory function. PMID- 12078496 TI - Enzyme-linked immunospot assay for detection of thioredoxin and thioredoxin reductase secretion from cells. AB - Oxidative stress response was determined in this study by enzyme-linked immunospot (ELISpot) assays for thioredoxin (Trx) and Trx reductase (TrxR). On exposure to oxidative stress, cells can launch a variety of defense mechanisms, including release of antioxidant proteins. The Trx system, consisting of Trx, TrxR, and NADPH, constitutes one of these cellular defense systems for maintenance of a healthy reduction-oxidation (redox) balance. Trx and TrxR are rapidly upregulated and released from monocytes, lymphocytes, and other normal and neoplastic cells on exposure. Secreted Trx and TrxR have proved to be eminent indicators of oxidative stress. Trx is a small, 12-kDa protein released through a leaderless pathway, whereas TrxR, which is a 116-kDa selenoprotein and required for regeneration of Trx, is secreted through the Golgi pathway. In this chapter we present a detailed laboratory bench protocol for enumeration of single cells secreting redox-active Trx and TrxR after oxidative stress exposure. Physiological stimuli (such as interferon gamma, lipopolysaccharide, interleukin 1, and CD23 ligation; and phorbol 12-myristate 13-acetate and ionophore) as well as UV light and hydrogen peroxide were used to generate oxidative stress, and some are presented in detail. The protocol includes a description of cell isolation, preparation, handling, and development of ELISpot plates, troubleshooting notes, presentation of results, statistical evaluation, and comments on alternative sources of materials and manufacturer Web addresses. We concluded that the ELISpot assay is a useful method for detection of single cells secreting the redox-active proteins Trx and TrxR after oxidative stress exposure. PMID- 12078497 TI - Functional evaluation of nonphagocytic NAD(P)H oxidases. PMID- 12078498 TI - Purification and assessment of proteins associated with nitric oxide synthase. PMID- 12078499 TI - Detection of redox sensor of ryanodine receptor complexes. PMID- 12078501 TI - Measuring reactive oxygen species inhibition of endothelin-converting enzyme. PMID- 12078500 TI - Redox control of 20S proteasome. PMID- 12078502 TI - Redox sensor function of metallothioneins. AB - In summary, the redox conversions of MT cysteines are likely to be the principal mechanisms for regulation of metal binding and release by this protein. Oxidative and/or nitrosative challenges can serve to promote metal ion release from MT to render their delivery to specific target proteins. It is tempting to consider the potential roles of MTs as redox sensors because of their high sensitivity to cysteine modification, as well as their potential to amplify signals by releasing multiple metal ions. In other words, MTs may act early in a biological signaling cascade that triggers metal-dependent biochemical and cellular responses. Alternatively, uncontrolled release of metals by excessive oxidative stress may contribute to metal toxicity. Because oxidative and nitrosative signaling is ubiquitous within cells, the physiological function of MT demands that efficient recycling of modified cysteines be operative. Little is known regarding the potential mechanisms for the regeneration of MT after oxidative/nitrosative modification, but they may involve endogenous dithiols, such as thioredoxin, and pharmacologically relevant dithiols, such as dihydrolipoate. PMID- 12078503 TI - SIR2 family of NAD(+)-dependent protein deacetylases. PMID- 12078504 TI - Mammalian two-hybrid assay showing redox control of HIF-like factor. PMID- 12078505 TI - Defining redox state of X-ray crystal structures by single-crystal ultraviolet visible microspectrophotometry. PMID- 12078506 TI - Model system for developing gene therapy approaches for myocardial ischemia reperfusion injury. PMID- 12078507 TI - Cloning and characterization of soluble decoy receptors. PMID- 12078508 TI - Using genetically engineered mice to study myocardial ischemia-reperfusion injury. PMID- 12078509 TI - Determination of redox properties of protein disulfide bonds by radiolytic methods. PMID- 12078510 TI - Transgenic model for the study of oxidative damage in Huntington's disease. PMID- 12078512 TI - Copper/zinc superoxide dismutase transgenic brain in neonatal hypoxia-ischemia. PMID- 12078511 TI - Heme oxygenase 1 transgenic mice as a model to study neuroprotection. AB - Bile pigments and CO are formed in the course of heme degradation by the isozymes and are biologically active moieties. In the course of heme degradation the chelated iron is also released. Heme and iron are prooxidants, whereas bile pigments are antioxidants. In addition, CO functions as a signal molecule and HO 2 may serve as an intracellular "sink" for NO. In the balance, the published data suggest that the HO system functions in cellular defense mechanisms. The methods described in this chapter can be used to assess the tissue/cell toxicity of chemicals in general, and as pertains to the defense activity of HO-1, specifically. PMID- 12078513 TI - Manganese superoxide dismutase transgenic mice: characteristics and implications. PMID- 12078514 TI - Tissue-specific knockout model for study of mitochondrial DNA mutation disorders. PMID- 12078515 TI - Antisense oligodeoxyribonucleotides: a better way to inhibit monocyte superoxide anion production? PMID- 12078516 TI - Transgenic shuttle vector assays for determining genetic differences in oxidative B cell mutagenesis in vivo. PMID- 12078517 TI - Crystal structures of oxidized and reduced forms of NADH peroxidase. AB - X-ray structural characterization of cysteine-sulfenic acid-containing proteins is one of the most defining approaches to characterizing this rapidly growing class of protein functional groups. Although outside the scope of this chapter, these structural analyses can lead to kinetic measurements in the crystal that allow intermediate states to be trapped, visualized, and studied. An understanding of the biochemistry of these reactive groups can be more fully gained by studying the localized protein environment in which these groups function. Increased perception of how elements of a protein can stabilize and contribute to modulation of function in these systems will allow novel means of enhancing or inhibiting function in important classes of protein molecules, including transcription factors and redox-regulated enzymes. PMID- 12078518 TI - Redox control of cell cycle-coupled topoisomerase II alpha gene expression. PMID- 12078519 TI - Chemokine expression in transgenic mice overproducing human glutathione peroxidases. PMID- 12078520 TI - Analysis of promoter methylation and its role in silencing metallothionein I gene expression in tumor cells. PMID- 12078522 TI - Reporter transgenes for study of oxidant stress in Caenorhabditis elegans. PMID- 12078521 TI - Functional genomics: high-density oligonucleotide arrays. PMID- 12078523 TI - Detection of DNA base mismatches using DNA intercalators. PMID- 12078524 TI - Deoxyguanosine adducts of tert-4-hydroxy-2-nonenal as markers of endogenous DNA lesions. PMID- 12078525 TI - Transcription-coupled repair of 8-oxoguanine in human cells. PMID- 12078527 TI - Nucleobase and 5'-terminal probes for DNA redox chemistry. PMID- 12078526 TI - Redox control of zinc finger proteins. PMID- 12078528 TI - Expressed sequence tag database screening for identification of human genes. PMID- 12078529 TI - Quantification of intracellular calcineurin activity and H2O2-induced oxidative stress. PMID- 12078530 TI - High-performance affinity beads for identifying anti-NF-kappa B drug receptors. PMID- 12078533 TI - The WHO Global Advisory Group on Nursing and Midwifery. PMID- 12078531 TI - Regulation of protein kinase C isozyme activity by S-glutathiolation. PMID- 12078532 TI - World Health Organization and global health policy. PMID- 12078534 TI - The systematic development of nursing interventions. AB - PURPOSE: To propose guidelines for systematic and progressive testing of nursing interventions. ORGANIZING FRAMEWORK: The National Institutes of Health has indicated four phases of clinical trials to guide research on treatments and interventions in the health sciences. Several important issues specific to nursing interventions need further clarification to enhance the interpretation of nursing clinical trials: (a) linking problem identification and nursing interventions, (b) articulating the content of interventions, (c) determining the strength and timing of interventions, (d) differentiating between efficacy and effectiveness research, and (e) incorporating multiple perspectives in intervention development. CONCLUSIONS: Systematic and progressive nursing intervention programs of research are needed to address these issues and to enhance the science and practice of nursing. PMID- 12078535 TI - Barriers to managing chronic pain of older adults with arthritis. AB - PURPOSE: To explore barriers to pain management experienced by older adults with arthritis, identify themes, and develop a theoretical model of relationships among the themes. DESIGN AND METHODS: Focus groups were used to collect data with 57 older-adult participants who had self-reported arthritis. Coding techniques consistent with grounded theory methodology were used for data analysis. FINDINGS: Nine themes were identified to describe barriers to chronic pain management experienced by older people with arthritis. The resulting model shows personal decision-making regarding the use of pain management methods. CONCLUSIONS: Findings provide direction for enhancing independent older people's ability to manage chronic pain. The nine categories of barriers show direction for education of both clients and health care providers, as well as for planning other types of needed support. The theoretical model shows the complexity of barriers experienced by older adults with arthritis. PMID- 12078536 TI - A concept analysis of uncertainty in illness. AB - PURPOSE: To examine the concept of uncertainty in illness and to propose an alternate model of uncertainty in the illness experience. ORGANIZING CONSTRUCT AND METHODS: Following a review of the literature, Morse's description of concept analysis by critically appraising the literature was used as a guideline in examining the concept of uncertainty. FINDINGS: Characteristics of the illness situation--ambiguity, vagueness, unpredictability, unfamiliarity, inconsistency, and lack of information--underlie the process of uncertainty. Three attributes of the concept of uncertainty were identified as probability, temporality, and perception. Loss of personal control is often erroneously equated with uncertainty. CONCLUSIONS: Uncertainty is a multidimensional concept that in its purest form is a neutral cognitive state and should not be mistaken for its emotional outcomes. To clarify the concept of uncertainty, further research is needed to determine the relationship of uncertainty to loss of control and psychosocial outcomes. PMID- 12078537 TI - College students' perceptions and practices of sexual activities in sexual encounters. AB - PURPOSE: To examine college students' understanding of safer sexual encounters, including expectation for sexual activities, planning for sexual encounters, and ideas about what constitutes "safe" sex. DESIGN: Exploratory design of college students' perceptions of and actual sexual activities that occurred in their last or most memorable sexual encounter. METHODS: Participants in one U.S. university completed three open- and closed-ended questions about their activities, as part of a larger questionnaire concerning contextual factors in their sexual encounters. A content analysis was conducted on data from open-ended questions. Encounters were categorized by risk for further analysis. FINDINGS: No significant differences were found in expectations regarding sexual activities between participants who had risky and safer encounters, except that participants who had safer encounters expected touching through clothing significantly less than did those who had risky encounters. Frequencies of sexual activities varied greatly between the two risk groups. No differences were found between the two groups concerning whether the encounter was planned or in expectation of sexual activities by planning of the encounter. The majority erroneously thought they had "safer" sex. The most reported reasons for believing a risky encounter was safer included the use of a hormone-based birth control method, use of a condom during vaginal sex only, and believing no sex had occurred in the case of oral sex. CONCLUSIONS: For these participants, planning of the encounter was not related to safer sexual activities and safer sexual activities were generally misunderstood. PMID- 12078538 TI - Sexual health practices of homeless youth. AB - PURPOSE: To describe the sexual health practices of homeless adolescents, examine relationships among variables in a conceptual model of sexual health practices, and determine direct and indirect effects of population characteristics, cognitive-perceptual factors, and behavioral factors on sexual health practices among homeless adolescents. DESIGN AND METHODS: Descriptive exploratory design. A survey was administered to a convenience sample of 414 homeless young men (244) and young women (170) aged 16-20 years, the majority of whom were Anglo American. FINDINGS: Thirty-five percent reported homosexual or bisexual orientation, and sexual orientation was reported as a reason for leaving home. Over half reported a history of sexual abuse and nearly one in four had been treated for gonorrhea. Safe-sex behaviors were related to age, time away from home, assertive communication, social support, future time perspective, connectedness, perceived health status, intentions to use condoms, and condom self-efficacy. A parsimonious model with good fit indicated that the only direct paths to safe-sex behaviors were future time perspective, intentions to use condoms, and self efficacy to use condoms, and the direct paths to sexual self-care behaviors were from assertive communication, social support, and self-efficacy to use condoms. CONCLUSIONS: As in other studies of homeless youth, respondents reported a high incidence of sexual abuse and homosexual and bisexual orientation. Their safe-sex behaviors were surprisingly similar to those of university students, were modestly related to cognitive-perceptual variables in the sexual health model, and might be amenable to brief culturally relevant interventions. PMID- 12078539 TI - Rational suicide in the terminally ill. AB - PURPOSE: To examine the current debate over the right to die specific to the rational suicide of terminally ill patients. METHODS: Literature was reviewed for information concerning historical end-of-life practices and the past acceptance of suicide. Another review showed philosophical opinions and perspectives that spanned from ancient Greece and the Roman Empire to modern philosophical discourse. A case study of a terminally ill woman who chose suicide is presented to apply the history and philosophy to nursing care today. CONCLUSIONS: As more nursing care is delivered in patients' homes, nurses will face this situation with increasing frequency. A call is made for organizations to provide guidelines for nursing practice. PMID- 12078540 TI - Differences in evidence-based care in midwifery practice and education. AB - PURPOSE: To examine site-specific differences in managing labor and to describe variations in intrapartum practice in the context of clinical midwifery education. DESIGN: Descriptive design based on secondary analysis of an existing data set collected to evaluate the intrapartum clinical experiences of nurse midwifery students. The data set included 498 records collected by midwifery students in 23 sites from 1995-1998. The unit of analysis was the intrapartum record. METHODS: Students used The American College of Nurse-Midwives Clinical data Set for Intrapartum Care to collect data during clinical experiences, including patient demographic data, risk factors, and specific care processes and interventions. FINDINGS: Significant variations were found among settings despite use of the same clinical guidelines. Discrepancies between theoretical preparation of students for clinical practice and the realities of clinical practice were noted. CONCLUSIONS: Despite a stated commitment to evidence-based practice, practice patterns varied significantly. Clinicians and educators need to find common ground for combining evidence-based theory with evidence-based practice. PMID- 12078541 TI - Institutional research responsibilities and needed infrastructure. AB - PURPOSE: To describe research grantee institutions' responsibilities to the funding agencies, research participants, and investigators, and the infrastructure and resources necessary for successful completion of funded research projects. ORGANIZING FRAMEWORK: Grants management includes investigator access to institutional and school infrastructure and resources, oversight and monitoring of the implementation of the research, and strategies to maximize research yield while stretching research dollars. CONCLUSIONS: Grantee institutions are responsible for overseeing the conduct of research, protecting rights of research participants, safety of study personnel, monitoring conflict of interest, fiscal management, access to data, and disseminating study findings. Central and school departments that provide needed services are important infrastructure supports. In addition to research space and investigator time, senior research mentors and research administrators are the most valuable resources for research success. PMID- 12078542 TI - Leadership research in business and health care. AB - PURPOSE: To summarize research on leadership in the health care and business literature and to identify the outcomes of leadership on individuals, groups, and organizations. METHODS: A computerized search and review of research studies was conducted in the health care and business literature from 1970-1999. Studies were categorized and analyzed according to participants, design, primary topic area, and effects or outcomes of leadership. FINDINGS: Most of the health care and business literature on leadership consisted of anecdotal or theoretical discussion. Only 4.4% (n = 290) of 6,628 articles reviewed were data-based. Further, the largest proportion of the research (120/290, 41.4%) was purely descriptive of the demographic characteristics or personality traits of leaders. Other studies showed the influence of leadership on subordinates (27.9%). Only 15 (5.2%) of 290 research articles include correlations of qualities or styles of leadership with measurable outcomes on the recipients of services or positive changes in organizations. CONCLUSIONS: Research on leadership in the health care and business literature to date has been primarily descriptive. Although work in the social sciences indicates that leadership styles can have a major influence on performance and outcomes, minimal transfer of this work to the health care system is evident. Limited research on leadership and health care outcomes exists, such as changes in patient care or improvements in organizational outputs. In this era of evidence-based practice, such research, although difficult to conduct, is urgently needed. PMID- 12078543 TI - Patients, nurses, and physicians have differing views of quality of critical care. AB - PURPOSE: To compare patient, nurse, and physician assessments of quality of care and patient satisfaction in selected critical care units. DESIGN: As part of a study of patient outcomes from critical care, data were collected between December 1991 and March 1993 from 489 patients, 518 nurses, and 515 physicians in 25 critical care units located in 14 hospitals in the U.S. Pacific Northwest. METHODS: Views of patient satisfaction and quality of care were measured using standardized instruments. All data were aggregated to the unit level (N = 25) and were normalized to a common scale for analysis. FINDINGS: Physicians rated quality of care higher than did either patients or nurses within the same critical care unit, and nurses had the lowest perceptions of quality. Nurses and patients had similar views of patient satisfaction, but physicians tended to overestimate patients' satisfaction. However, physicians', nurses', and patients' scores varied considerably within and between units. Physicians' and nurses' views of quality and patient satisfaction were strongly related to processes such as MD-RN collaboration and outcomes such as nurses' job satisfaction. CONCLUSIONS: Patients, nurses, and physicians viewed quality of care and patient satisfaction differently. Nurses' and physicians' perspectives were more related to their views of the work environment than to organizational factors, patient characteristics, or commonly used outcome measures. PMID- 12078544 TI - How nurses learn advocacy. AB - PURPOSE: To describe how nurses develop the skill of advocating for patients. DESIGN & METHODS: Hermeneutic phenomenological research methods. Both reserve and active U.S. Army nurses who cared for patients associated with the military operation in Bosnia were individually interviewed to gain an understanding of their experiences of advocating for patients and how they developed their advocating practices. FINDINGS: The constitutive pattern identified was Developing Advocating Practices. The themes comprising this constitutive pattern were Who I Am, Watching Other Nurses Interact with Patients, and Gaining Confidence. CONCLUSIONS: For the nurses in this study, developing advocating practices was more haphazard and situationally dependent than methodically taught in their nursing education programs. Nurses would have a stronger foundation in advocacy in nurse educators consciously teach advocacy and if nurse administrators support new graduates' advocacy in the work environment. PMID- 12078545 TI - Early risk indicators of substance abuse among nurses. AB - PURPOSE: To investigate early risk factors that led to substance-related disorders and to predict group differences between substance-impaired (SI) and nonimpaired (NI) registered nurses. ORGANIZING CONSTRUCT: Donovan's multifactorial model of impairment, and Rogers' Science of Unitary Human Beings. METHODS: Data were gathered from 100 previously SI and 100 NI nurses located through use of the Internet. Three questionnaires were used: the Zuckerman Sensation Seeking Scale (ZSSS), the Efinger Alcohol Risk Survey (EARS), and the Children of Alcoholics Screening Test (CAST). FINDINGS: Independent t-test scores showed the two groups differed significantly on all three instruments' total scores. Discriminate analysis indicated a correct prediction of 87% for SI and 95% for NI nurses, with an overall rate of 91%. EARS scores were the best predictor of nurses with substance-related disorders (.99), followed by ZSSS (.44) and CAST (.42) scores. CONCLUSIONS: The three variables indicate early risk factors for substance-abuse impairment. Identification of nurses at risk for impairment will allow for earlier intervention and possible prevention. Methods to reduce the number of modifiable risk factors are recommended. PMID- 12078546 TI - [Diaphragmatic hernia from 1961 to 2000--incidence, prenatal diagnosis and prevalence according to maternal age]. AB - OBJECTIVE: Presentation of diaphragmatic hernia in the CR in 1961-2000. Analysis of prevalence of these defects in different groups by maternal age. DESIGN: Retrospective demographic-epidemiological study. SETTING: Institute for the Care of Mother and Child, Prague. METHODS: The authors used data from nationwide registers of birth defects in neonates kept in the Institute of Health Information and Statistics of the CR during 1995-2000. Epidemiological analysis of the incidence of diaphragmatic hernia in the CR during 1961-2000 in newborn infants. Mathematical statistical analysis of prevalence of these defects by maternal age using the method of 95% confidence interval of probability. RESULTS: In the CR during 1961-2000 1321 cases of diaphragmatic hernia were recorded. The mean incidence of this defect during the entire period was 2.32 per 10,000 live born infants incl. prenatally diagnosed cases and terminated pregnancies. Including those it was 2.37 per 10,000 live born infants. CONCLUSION: The authors elaborated the mean incidence during the investigation period and in individual years in born children as well as prenatally diagnosed cases. Also an analysis of the prevalence of these defects by maternal age was made using the method of 95% confidence interval. The authors found a significantly higher risk of development of this defect in foetuses of women aged 38, 42 years or older as compared with the population average. However an unequiwocally increased risk associated with more advanced age was neither confirmed nor ruled out. PMID- 12078547 TI - [Congenital renal defects from 1961 to 2000--incidence, prenatal diagnosis and prevalence according to maternal age]. AB - OBJECTIVE: Presentation of agenesis/hypoplasia of kidneys and cystic kidneys in the Czech Republic during 1961-2000. Analysis of prevalence of these defects in different groups by maternal age. DESIGN: Retrospective demographic epidemiological study. SETTING: Institute for the Care of Mother and Child, Prague. METHOD: The author used data from the nationwide register of birth defects in neonates, kept by the Institute of Health Information and Statistics in the CR during 1990-2000. Epidemiological analysis of the incidence of agenesis/hypoplasia of the kidneys and cystic kidneys in the CR during 1961 to 2000 in cases diagnosed prenatally and in born children. The authors made a mathematical-statistical analysis of the prevalence of these defects by maternal age, using the method of calculation of the 95% confidence interval of probability. RESULTS: In the CR there were during the period from 1961-2000 1093 registered cases of agenesis/hypoplasia of the kidneys and 1370 cases of cystic kidneys. The mean incidence during the entire period was 1.81 in agenesis/hypoplasia of the kidneys and 2.26 in cystic kidneys per 10,000 live born infants. After adding prenatally diagnosed cases and pregnancies terminated on that account it was 2.05 a 2.58 per 10,000 live born infants. CONCLUSION: The investigation of these inborn defects did not confirm nor rule out statistically a higher risk of women older than the population average. PMID- 12078548 TI - [Integration of prenatal screening for Down syndrome in the first and third trimester (proposal for a screening protocol]. AB - OBJECTIVE: Proposal of antenatal Down syndrome screening protocol using integration of first and second trimester results. DESIGN: Review. SETTING: Antenatal screening centre GENNET, Prague. METHODS: Search of relevant articles, software analysis and pilot implementation. RESULTS: Screening of Down syndrome (DS) is divided into two phases: at first phase combined DS risk is calculated using pregnancy plasma protein-A (PAPP-A) at 9th-11th gestational week and ultrasound fetal nuchal translucency (NT) at 11th-13th week with risk cut-off 1/100 (about 2 per cent of screened). Positive mothers are counselled immediately after NT measurement: First phase negative mothers continue to second phase where PAPP-A and NT values are integrated with second trimester screening results. Protocol is supported with special software (Obstetrician's Screening Guide-OSG). CONCLUSION: At proposed protocol the information on significant first trimester DS risk is not withheld to second trimester and mother can choose early prenatal diagnosis. Expected detection rate is near 90 per cent at 5 per cent false positive rate. PMID- 12078549 TI - [Fructose and glucose in follicular fluid and serum of women undergoing stimulation in an in vitro fertilization program]. AB - OBJECTIVE: To find out if there is some amount of fructose in the follicular fluid of IVF stimulated patients. If so then to compare fructose and glucose levels with IVF outcome, type of stimulation and infertility factors. DESIGN: Retrospective study. SETTING: Department of Gynaecology and Obstetrics, Charles University, the Czech Republic. METHODS: 202 women undergoing IVF treatment since 11/1999 till 02/2001 were evaluated. We measured a level of fructose and glucose in a sample of serum and follicular fluid (FF) obtained at ovum pick-up. Then the results were compared with the IVF outcome, the length of stimulation and infertility factors. The control group consists of 23 women with a pure andrologic factor of infertility. RESULTS: At the day of pick-up the serum concentration of glucose and fructose taken on an empty stomach is on average 5.43 +/- 1.35 mmol/l and 25.05 +/- 9.26 micrograms/ml compared to 5.26 +/- 1.00 mmol/l and 23.69 +/- micrograms/ml in control group. The concentration of both glucose and fructose in FF correlates with their serum level. The FF concentration of glucose is on average 3.75 +/- 1.32 mmol/l (70% of serum concentration) and fructose 11.34 +/- 5.29 micrograms/ml (48% of serum concentration) by comparison with 3.78 +/- 0.91 mmol/l (73%) and 12.37 +/- 7.06 micrograms/ml (53%) in control group. CONCLUSION: There is a possible influence of follicular fluid saccharides like glucose and fructose in process of fertilization. We derived this hypothesis from studies evaluating the role of saccharides in endometrial secretion, cervical mucous and seminal plasma. We assumed that patients with a successful IVF cycle have a lower fructose concentration in FF. The clinical pregnancy is really mildly lower but this difference is not statistically significant. We confirmed FF fructose is in close correlation with its serum concentration. But we did not confirm the FF glucose and FF fructose concentration is related to the IVF outcome. PMID- 12078550 TI - [Hydrogen expandable implants in the treatment of stress incontinence in women--a pilot study]. AB - OBJECTIVE: The aim of the study was to evaluate the use of new-type implants for the treatment of female urinary stress incontinence. The patients with intrinsic sphincter deficiency were also included, the other were after previous surgery for GSI. DESIGN: Original article. SETTING: Departments of Obstetrics and Gynaecology, Faculty Hospital Na Bulovce and General Faculty Hospital, 1. Faculty of Medicine, Charles University, Prague, Institute of Macromolecular Chemistry, Czech Academy of Science, Prague. METHODS: We operated 9 patients, two of them with type III (McGuire) urinary stress incontinence the rest after operations for GSI in general anesthesia. Six to nine hydrogel swelling implants were introduced into the submucosa of proximal urethra under the transrectal ultrasound guidance. We evaluated the effect 6 to 9 months after the operation. RESULTS: Two patient were dry (negative stress-test and pad-weight test), one patient had substantial improvement, 4 had mild improvement and 2 did not notice any change. We found de novo urge symptoms in four patients, two of them were of the motor type. CONCLUSION: The new type of implants reached the success rate of other currently used materials. The improvement of stress incontinence was on the cost of de novo urge symptoms in some patients. The urgency was successfully managed with spasmolytic drugs. PMID- 12078551 TI - [Diverticula of the female urinary tract--still a problem in urogynecology]. AB - OBJECTIVE: An importance of knowledge of etiology, pathogenesis, diagnosis, differential diagnosis and treatment of female urethral diverticulum in urogynaecological practise. DESIGN: A review article. SETTING: Obstetrics and Gynaecology Department and Urology Department, Charles University 2nd Medical School and Teaching Hospital Motol, Prague. SUBJECT: Often unrecognized or incorrectly diagnosed, urethral diverticulum may be completely asymptomatic or may produce significant symptomathology, such as recurrent urinary tract infections and irritative or obstructive complaints. A full history and careful physical examination are the first steps in screening. Diagnosis is usually made by voiding cystourethrography, positive pressure urethrography, urethroscopy or introital ultrasonography. Complete excision through the anterior vaginal wall is the most successful treatment modality with minimum postoperative complications. Concomitant bladder neck suspension can be performed in women with diverticulum and documented stress urinary incontinence. CONCLUSION: Urethral diverticula may mimic other pelvic floor disorders and result in diagnostic delay. It is partly due to a lack of awareness among clinicians and partly because the condition overlaps the traditional territories of gynaecologists and urologists. The collaboration of both specialists in the diagnostics and treatment of female urethral diverticula is recommended. PMID- 12078552 TI - [Arterial diseases in women using combined hormonal contraceptives]. AB - OBJECTIVE: To determine the association between myocardial infarction and cerebral stroke and use of combined oral contraceptives. DESIGN: Review of literature. SETTING: Department of Obstetrics and Gynaecology, Hospital of Merciful Brothers, Brno, Czech Republic. METHODS: Identification of methodologically sound studies able to address the topic. Studies were identified by Medline database search. RESULTS: Five the most recent and important studies were the main source of information (Oxford Family Planning Association Contraception Study 1984, Royal College of General Practitioners' Study 1994, WHO Collaborative Study of Cardiovascular Disease and Steroid Hormone Contraception 1996, 1997, Transnational Study on Oral Contraceptives and the Health of Young Women 1997, Myocardial Infarction and Oral Contraceptives Study, 1997). Generally there was no evidence of a significantly increased risk of arterial wall disease in healthy non smoked users younger than 35 years. No difference between second and third generation oral contraceptives on risk of arterial wall disease were found. In the most of cases of myocardial infarction or stroke one or more risk factor were identified. Two of the most relevant risk factors are smoking and the absence of blood pressure control. CONCLUSION: The risk of arterial cardiovascular system diseases seems not to be increased at no-risk users (healthy non-smoker younger than 35 years). PMID- 12078553 TI - [An unusual tumor of the oral cavity in a fetus and prenatal ultrasonography- case report]. AB - OBJECTIVE: The tumours and pseudotumours of the foetal face are very rare diseases. Basic method for their prenatal diagnosis is ultrasound. The authors describe the case of diagnosed orofacial pseudotumour--sublingual cyst. DESIGN: Case report introduces some rules for ultrasound investigation of foetal face. SETTING: Department of Clinical Genetics and Foetal Medicine, University Hospital Olomouc. SUBJECT AND METHOD: The cyst of oral cavity was diagnosed by ultrasound in second trimester of gestation. It was repeatedly punctured during pregnancy and the content was cytologically examined. Exstirpation was performed after delivery. Histological etiology--dysontogenetic cyst--was established. CONCLUSION: Prenatal ultrasonographical diagnosis of the oral pseudotumour was completed by puncture procedures. This invasive way verifies anatomical conditions, enables histological investigation and warrants the swallowing act which is the prevention of polyhydramnion. PMID- 12078554 TI - [Recommendations for diagnosis and therapy of menstrual cycle disorders]. PMID- 12078555 TI - [Sexual life of women with hyperphenylalaninemia]. AB - OBJECTIVE: Characteristics of sexual life in women with hyperphenylalaninemia. DESIGN: Anamnestic study. SETTING: Department of Obstetrics and Gynaecology and Institute of Inherited Metabolic Disorders of General Faculty Hospital and 1st Medical Faculty of Charles University in Prague, Department of Clinical Biochemistry, Haemathology and Immunology of Hospital Na Homolce in Prague. METHODS: Characteristics of sexual life were studied in a group of 16 women with hyperphenylalaninemia. RESULTS: One fourth of the women lives without sex. In the rest of them sexual function is normal. CONCLUSION: One fourth of the women stays out of reproduction. PMID- 12078556 TI - [Legal and ethical problems in assisted reproduction--case report]. AB - OBJECTIVE: The objective of the study is to evaluate legal and ethical aspects of the assisted reproduction in the Czech Republic. These aspects are described in the context of the concrete case. DESIGN: Case report. SETTING: Department of Social Medicine, Medical and Health Care Administration Faculty of the Masaryk University, Brno and Centre of the Reproductive Medicine Zlin. SUBJECT AND METHOD: The description of the concrete case that happened in the Czech Republic. A widow asked for the artificial insemination using the sperm of her deceased husband. CONCLUSION: It was concluded that there is absolutely insufficient legal regulation of the assisted reproduction techniques, it does not respond to the development of the medical knowledge and it responds insufficiently to the ethical dilemmas concerning the assisted reproduction. PMID- 12078557 TI - Is a double cheeseburger really public health enemy #1? PMID- 12078558 TI - NP roles in women's health management. PMID- 12078559 TI - Reshaping recruiting. Strategies for increasing diversity in academic nursing programs. PMID- 12078560 TI - Pre-diabetes warnings going out. Diabetes epidemic on the uprise; pre-diabetes going undiagnosed. PMID- 12078561 TI - Undersize infants score higher IQ if breastfed exclusively. PMID- 12078562 TI - Gynecologic oncologists making strides in women's cancer care. PMID- 12078563 TI - Infectious childhood heart disease exposed. PMID- 12078564 TI - More Americans facing blindness. PMID- 12078565 TI - Lycopene may contribute to heart health. PMID- 12078566 TI - Bioterrorism & pregnant women. Promoting access to care in times of crisis. PMID- 12078568 TI - Spinal cord injuries. Nurses can help affected women & their families achieve pregnancy birth. PMID- 12078567 TI - Confronting cervical cancer. Screening is the key to stopping this killer. PMID- 12078569 TI - Infant security education. A multidisciplinary approach. PMID- 12078570 TI - Breast reduction surgery. What every woman needs to know. PMID- 12078571 TI - Managing difficult conversations. Novice managers face fast learning curve. PMID- 12078572 TI - Culturally relevant genetic counseling. Nurses play a critical role in helping women & families. PMID- 12078573 TI - Surviving thyroid cancer. This silent cancer affects both young & old women. PMID- 12078574 TI - A defining time for dental hygiene. PMID- 12078575 TI - Information-seeking behaviors of U.S. practicing dental hygienists and full-time dental hygiene educators. AB - PURPOSE: This study compared the frequency and variety of information-seeking behaviors used by dental hygiene practitioners and dental hygiene educators to answer practice-related questions. Personal demographic characteristics also were compared between the two groups to determine the characteristics of those with a high level of computer-based information-seeking behaviors. METHODS: A self administered questionnaire was sent to a national computer generated random sample of 300 practicing dental hygienists and 300 dental hygiene educators. Responses were received from 63% of the practicing dental hygienists and 78% of the educators. RESULTS: Dental hygiene practitioners and dental hygiene educators both used traditional methods of seeking information. However, dental hygiene educators used a higher frequency and wider variety of sources than dental hygiene practitioners, including innovative sources such as the Internet, computer databases and online discussion groups. Dental hygiene educators almost universally reported some use of the Internet and computer databases--97% and 91%, respectively. Meanwhile, 50% of dental hygiene practitioners used the Internet and 25% used computer databases at least once for practice information. Online discussion groups were used by 50% of dental hygiene educators but by only 13% of dental hygiene practitioners. Level of education was the demographic characteristic that was most associated with use of computers to answer practice related questions. CONCLUSIONS: Since use of computer sources for information were defined as innovative, more dental hygiene educators were classified in early adopter categories, while greater numbers of dental hygiene practitioners were classified in later adopter categories. Group demographics were examined to determine variables that predicted adoption. Level of education predicted the majority of variation and as a whole, showed the strongest statistical differentiation between adopter categories. PMID- 12078577 TI - Oral health issues among migrant farmworkers. AB - PURPOSE: The purpose of the study was to determine utilization patterns of dental services, unmet dental needs, access to care barriers, and oral health behaviors as perceived by migrant farmworkers at a rural southern Illinois farmworker health clinic. METHODS: Two bilingual dental hygiene students and one member of the local Hispanic community verbally administered a 26-item survey questionnaire to 119 migrant farmworker clients at a health center as they waited to receive care. RESULTS: Utilization results showed that 51% of those surveyed had not sought oral health care in the previous year, citing absence of pain or discomfort as the primary reason. Forty-one percent reported seeking oral health care on a yearly basis, while 42% only sought care when in pain. Primary services received were examinations, prophylaxes, and restorations. Having received brushing instructions was reported by 58%, while 45% had received instructions on flossing. Barriers to care were reported as limited clinic hours (57%), high fees (33%), and lack of transportation (17%). Most respondents reported regular brushing habits, but only 11% used floss daily, 38% occasionally, and 52% didn't use it at all. Only 7% reported smoking. Meanwhile, bleeding gingiva was reported by 50%, swollen or tender gingiva by 37%, and tooth loss by 49%. CONCLUSIONS: The majority of migrant farmworkers in a southern Illinois community reported access to care barriers, and having never or episodically received dental services. Nearly half reported signs of periodontal disease. PMID- 12078576 TI - Prevalence of carpal tunnel syndrome and upper extremity tendinitis among dental hygienists. AB - PURPOSE: This study was undertaken to determine the prevalence of carpal tunnel syndrome (CTS) and upper extremity (UE) tendinitis among dental hygienists. METHODS: In a cross-sectional study dental hygienists (n = 305) were screened using sensory nerve conduction, a focused physical examination of the UE, and a symptom questionnaire. CTS was diagnosed if the subject had slowing of the median nerve at the wrist and symptoms of numbness, tingling of pain in the median distribution. Localized tendinitis of the UE was diagnosed if the subject had focal symptoms and associated findings on physical examination. RESULTS: Three percent of the participating dental hygienists were diagnosed with CTS. Thirteen were diagnosed with shoulder tendinitis, while 6% had a tendinitis of the elbow and 7% had tendinitis of the hand or wrist. Twenty-eight percent had a diagnosis of some UE tendinitis or CTS. CONCLUSIONS: The prevalence of hand and finger symptoms in the dominant hand among dental hygienists in this study was high, but the prevalence for CTS was nearly the same as the general population. There was a high rate of UE tendinitis noted within this population. PMID- 12078578 TI - A comparison of dental caries and tooth loss for Iowa prisoners with other prison populations and dentate U.S. adults. AB - PURPOSE: When one compares National Health and Nutrition Examination Survey III (NHANES III) data to previous national surveys it appears that oral health in the United States has improved. The purpose of this study was to determine if this trend holds for prisoners in Iowa, specifically in regard to number of decayed teeth, number of decayed surfaces, and number of missing teeth. METHODS: This original research is a report of cross-sectional data regarding untreated decay and missing teeth. A representative sample of inmates newly admitted to the Iowa Medical Classification Center (IMCC) between June and December 1998 was selected. Oral health data were collected from records obtained as the result of dental treatment screening examinations conducted by the IMCC staff dental hygienist. Demographic data were obtained from questionnaires. The oral health status of this Iowa inmate population was compared with the status of previously studied samples of inmates throughout the nation using descriptive and parametric statistics and with dentate, noninstitutionalized U.S. adults using descriptive, non-parametric, and parametric statistics. RESULTS: Of the 174 study participants, 149 were male and 25 were female. On average, male inmates had 7.09 decayed teeth, 15.3 surfaces of decay, and 4.07 missing teeth. The female inmates averaged 5.56 teeth with untreated decay, 14.4 surfaces of decay, and 5.12 missing teeth. Findings showed that male Iowa inmates have more untreated decay than previously studied samples of inmates throughout the nation. IMCC Caucasian male inmates had 1.6 to 7.8 times as many untreated decayed teeth as previous inmates, while African American inmates showed 1.2 to 3.4 times as many untreated decayed teeth. The Iowa male inmates had fewer missing teeth than most of the inmate comparison groups. Compared with dentate, noninstitutionalized U.S. adults, the IMCC inmates, male and female combined, had 8.4 times the amount of untreated decay but similar numbers of missing teeth. CONCLUSIONS: Newly admitted male and female inmates were disparate from the general dentate U.S. adult population in terms of untreated decay. This also was found for IMCC male inmates compared to previous studies of male inmates. Inmates of both genders at IMCC did not appear to be disparate with the comparison population with regard to number of missing teeth. PMID- 12078579 TI - Hemolytic bacteria in water from the dental air-water syringe. AB - PURPOSE: The purpose of this pilot study was to determine whether aquatic bacteria expressing hemolysis, a classical virulence factor of pathogenic bacteria, could be found in water flowing from the dental air-water syringe (AWS). METHODS: Water samples were collected from the AWS in each of five dental operatories, and a control sample was collected from the cold-water tap in the sink adjacent to one of the dental operatories. Water samples were plated on NWRI agar to enumerate aquatic heterotrophic bacteria and on Tryptic Soy Blood (TSB) agar to enumerate hemolytic bacteria. After incubation, plates were counted to determine total colony-forming units per mL of AWS water. In addition, TSB plates were examined for hemolytic colonies, which were counted if present. RESULTS: AWS water samples contained an average of 9.4 x 10(4) colony-forming units cfu/mL on NWRI agar, and 1.3 x 10(4) cfu/mL on TSB agar. The control sample from the cold water tap contained 2.5 x 102 and 50 cfu/mL on NWRI and TSB agars, respectively. Four out of five AWS water samples contained hemolytic bacteria, with an average hemolytic count of 3.2 x 10(3) hemolytic cfu/mL on TSB agar. No hemolytic bacteria were detected from tap water. Most hemolytic organisms exhibited alpha hemolysis although beta-hemolytic colonies were seen. CONCLUSIONS: AWS water can contain hemolytic bacteria--such bacteria express a virulence factor and therefore may be regarded as potential pathogens. According to a Medline search conducted April 23, 2002, this is the first report of hemolytic bacteria from dental unit water. PMID- 12078580 TI - Prioritization of the National Dental Hygiene Research Agenda. AB - The profession of dental hygiene has made considerable progress over the past 30 years toward developing a unique body of knowledge for guiding education, practice, and research. The 1993-1994 American Dental Hygienists' Association Council on Research published the first national dental hygiene research agenda in 1994. The 1994 research agenda focused dental hygienists' research efforts; however, publication of two national reports--the Surgeon General's Report on Oral Health, and Healthy People 2010--have made it necessary to revisit the research agenda. After considering input from participants in the Fourth National Dental Hygiene Research Conference and evaluating the Surgeon General's Report, the 2000-2001 Council on Research has established recommendations for the prioritization of the 1993-1994 research agenda. This report outlines for readers the rationale for the proposed recommendations. PMID- 12078581 TI - [Genetic syndromology. Introduction to a series]. AB - Syndromology belongs to diagnostic methods based on the analysis of phenotypic (clinical or anatomical--dysmorphics) features, which occur very often together and have a common etiology (e.g. teratogenic embryopathy, numerical and structural chromosomal aberrations or gene mutations). In the phenotype analysis important appear so-called signal features, which enable to narrow the range of possible disorders for the differential diagnosis of the assumptive diseases. PMID- 12078582 TI - [Hyperbaric oxygen therapy in athletic injuries]. AB - Hyperbaric oxygen therapy (HBO2) may play an important role in management of sport injuries: in football and hockey players, and after other sport activities. When HBO2 is applied together with physical therapy and rehabilitation procedures, it can accelerate the healing and thus to decrease the costs of treatment. PMID- 12078583 TI - [Ethical, psycho-social, legislative and economic aspects of surgical treatment of the hypoplastic left heart syndrome]. AB - Hypoplastic left heart syndrome (HLHS) represents a lethal congenital heart disease, which used to be considered inoperable. The improvement of results of cardiac surgery in neonates improved also the prognosis of patients with this heart lesion. HLHS, however, continues to be an unresolved problem from ethical, psychological, social and economical points of view. Opinions regarding the care for these patients depend on economical, political, cultural and religious factors. Based on recent experience it is evident, that HLHS is an operable disease in a child, who without surgery would otherwise die. Approximately 40% of patients with HLHS can live 15 years after surgery. The long-term prognosis remains, however, unclear. Because of the need of demanding care for a patient with HLHS, it is necessary to see also the quality of life and hear the opinion of parents. Psychomotor development and the quality of life may be altered by complications. An individualized approach in fetuses and neonates with HLHS on the basis of prenatal diagnosis, analysis of risk factors, provision of detailed informations to parents and indications for surgery in neonates having chance for an acceptable result is considered to be the most acceptable solution. In the future it will be necessary to concentrate the research on prenatal diagnostics, treatment and postoperative follow-up including psychomotor development and quality of life. In the preparation of legislative, which would reflect more sensitively the contemporary possibilities of medicine and at the same time regard the problems of health care organization, as well as social and economic aspects, narrow interdisciplinary cooperation will be required. PMID- 12078584 TI - [Predictive features of positron emission tomography after two cycles of induction therapy in malignant lymphoma]. AB - BACKGROUND: Positron emission tomography (PET) is a modern functional imaging method, recently introduced to clinical oncology. The aim of our study was the evaluation of prognostic value of PET performed in malignant lymphoma patients after two cycles of chemotherapy. METHODS AND RESULTS: From 9/99 to 11/00 PET was performed in 37 patients with malignant lymphoma (9x m. Hodgkin, 21x HG + IG-NHL, 7x LG-NHL; 26x new diagnosis, 11x relaps of disease). Freedom from progression interval (FFP) and overall survival (OS) were evaluated. Attenuation corrected PET imaging was done by dedicated ECAT EXACT PET scanner from base of the skull to the upper thighs 1 hour after intravenous administration of 18-FDG (7.6 +/- 1.3 MBq/kg). Statistical analysis was done using Kaplan-Meier method. Significance of differences between groups was determined by log-rank test on the level of 5%. After the induction therapy, 30 patients were in complete remission, 3 patients in partial remission and in 4 cases progression of disease were observed. Progression of disease was seen in 4 patients. Median follow up of living patients was 7 months (1-13 months) from the end of therapy. Progression, resp. relapse of disease occurred in 13 patients during this period, two patients died. PET performed after the second course of therapy was positive in 18 patients and negative in 19 patients. Two progressions, resp. relapses of disease were documented in PET-negative group and 11 in PET-positive group. FFP was significantly different in PET positive and PET negative groups (p < 0.05). The negative and positive predictive values of PET for malignant lymphoma relapse or progression were estimated 89%, 63% respectively. CONCLUSION: Regardless the short follow-up period, our preliminary results reflect very good prognostic value of PET performed after the second course of chemotherapy in malignant lymphoma patients. PMID- 12078585 TI - [Effect of amifostine on reducing acute toxicity of megachemotherapy of tumors in children]. AB - BACKGROUND: Amifostine (WR-2721, Ethyol) is a chemoprotective agent. There is little experiences with amifostine application in megachemotherapy in children. We evaluated amifostine effect on the reduction of the acute toxicity. METHODS AND RESULTS: Retrospective comparison of patients who received amifostine with the control group (72 vs. 72). Amifostine 750 mg/m2 was given 15 minute before cytostatic dose and regularly each eight hours if we administered cytostatics continuously. Megachemotherapy schedule included melfalan, carboplatin, cyklophosphamid, vepesid, busulfan, thiotepa and karmustin. Type of graft: peripheral stem cells 56 vs. 29, bone marrow 8 vs. 30, combination 8 vs. 13. Nonhematological toxicity: mucositis p = 0.047, hepatotoxicity p < 0.001, nephrotoxicity p = 0.005. Hematological toxicity: engraftment D + 12 vs. D + 15 (p < 0.001), amount of erythrocyte transfusions 3 vs. 6 (p < 0.001), platelet transfusions 7 vs. 9 (p = 0.06), days when number of platelets reaches 20,000 without substitution D + 15 vs. D + 22 (p < 0.001). The only statistically difference was in the in total amount of platelets (p = 0.032), when we calculated patients, who received peripheral stem cells. Number of hospitalization days 14 vs. 18 (p = 0.016), days with antibiotics 14 vs. 18 (p = 0.016), number of febrile days 6 vs. 7 (p = 0.51). CONCLUSIONS: Amifostine reduces mucosal, liver and kidney damage. The graft type could affect hematological results. PMID- 12078587 TI - [IKEM (The Institute for Clinical and Experimental Medicine)--undergraduate and postgraduate education]. PMID- 12078586 TI - [A system for monitoring quality, outcomes and health care costs]. AB - Specialised form of database named DataCube for assessment of quality, outcomes and costs of the healthcare in cardiac surgery was built at the Institute of Clinical and Experimental Medicine (IKEM). Risk stratification of patients was done using Bayes predictive model. Such prediction well correlated with calculated costs. The results achieved promise the system to become an important decision making tool, which may improve identification of high risk patients. PMID- 12078588 TI - Hyphenated dentistry. PMID- 12078589 TI - Hyphenated dentistry. PMID- 12078590 TI - Implant dentistry. PMID- 12078591 TI - The Japanese market for dental implants: executive summary. PMID- 12078592 TI - Implant restorations and their place in the dental practice. PMID- 12078593 TI - Immediate restoration of multiple tooth implants for aesthetic implant restorations. AB - The immediate restoration or loading of dental implants has been an intense area of clinical trial and research in the field of dental implantology over the last several years. The ability to temporize implants that are placed and prevent the use of a removable transitional appliance is appealing to not only patients, but dentists as well. Additionally, the placement of implants into immediate extraction sockets, and the immediate restoration of those implants, is showing excellent success rates clinically. Enhancement of the healing phase using platelet rich plasma seems to be a key contributor to the healing and maturation rate of both the hard and soft tissues. This article will review the immediate restoration of implants placed into multiple extraction sockets and the use of platelet rich plasma to enhance the healing phase. PMID- 12078594 TI - Immediate loading of osseotite implants: two-year results. AB - The high success rate of dental implants has changed the quality of life for many patients. Immediate loading finds its application in some clinical cases and certainly adds another modality of treatment for the implant patient. Starting with a couple of implants placed in the mandible and immediately loaded within 72 hours after surgical placement with an overdenture, this concept evolved to immediately loading multiple implants in both the maxilla and the mandible. In this investigation, 11 consecutive patients were treated with 87 screw-shaped endosteal acid-etched, Osseotite implants (3i, Implant Innovations, Inc., Palm Beach Gardens, FL) between June 1998 and June 1999. Two mandibular and two maxillary cases received screw-retained provisional prostheses the day of surgery. Three mandibular and four maxillary cases were loaded 48 hours after surgery with the final screw-retained, porcelain-fused-to-metal prostheses. All implants were followed for 2 to 3 years. Follow-up consisted of clinical as well as radiographic examination. All implants were successful. There was no implant mobility or periimplant radiolucency. The bone level was measured at the 12th and 24th months. The average radiographic bone level from the implant platform to the first bone-to-implant contact was 0.654 mm at the 12th month and 0.946 mm at the 24th month. We conclude that a high success rate can be achieved when implants with a hybrid surface, machined/acid-etched, are immediately loaded within 48 hours after surgical placement in the maxilla and the mandible. PMID- 12078596 TI - SwissPlus Implant System, Part 1: Surgical aspects and intersystem comparisons. AB - Implant survival in poor-quality bone continues to pose a significant clinical challenge to dentists. The SwissPlus System comprises one-piece, straight and tapered implant designs with self-tapping, apical threads, and a microtextured surface on the intraosseous portion of the implant bodies. Although both designs are indicated for all ranges of bone density, Tapered SwissPlus features double lead threads and a soft-bone surgical protocol designed to enhance initial mechanical stability at the time of placement. This paper presents an overview of the SwissPlus System with emphasis on the surgical aspects. Presented test data also illustrate intersystem compatibility and differences between the straight SwissPlus and ITI synOcta implants. PMID- 12078595 TI - Fracture of dental implants: literature review and report of a case. AB - Fracture of dental implants is a rare phenomenon with severe clinical results. In this article, the literature is reviewed and various causative factors that may lead to fracture are presented. Galvanic activity has not been mentioned before as a possible cause for implant fracture, yet, it can occur at the level of contact with the superstructure. This is illustrated by the case of a titanium implant restored with a non-precious porcelain-fused-to-metal cemented crown that fractured 4 years after loading. The radiographs show alveolar bone resorption around the fixture. Metallurgical analysis of the implant indicated that the fracture was caused by metal fatigue and that the crown metal, a nickel-chromium molybdenum alloy, exhibited corrosion. These findings suggest a new explanation for implant fractures; cytotoxic nickel ions, leaching from the base metal alloy may cause bone resorption. This in turn leads to increased mobility, facilitating washout of the luting cement. Contact of the base metal with titanium in the presence of oral fluids produces galvanic currents that hasten corrosion and leaching out of nickel ions, thus leading to further bone resorption. Loss of bone support allows lateral bending moments that cause metal fatigue, eventually leading to fracture. Therefore, good treatment planning and appropriate case selection might have prevented this fracture. Furthermore, the use of nonprecious metal alloy for the crown's infrastructure had further contribution to the chain of events that led to the implant's fracture. PMID- 12078597 TI - Rehabilitation of the edentulous maxilla and mandible with fixed implant supported restorations applying immediate functional loading: a treatment concept. AB - The rehabilitation of the complete edentulous arch with fixed implant-supported bridges can be a particular challenge for the practitioner. In the treatment concept presented, each jaw is provided with sufficient implants to realize fixed restorations under immediate (2-3 days after surgery) functional loading. After a healing time of 40 to 60 days, the metal/resin transitionals are exchanged for definitive metal/porcelain bridgework under gnathological aspects. Fixed implant supported bridges correspond to the arches of natural teeth and can reveal better results in function, phonation, and aesthetics than removable prostheses. PMID- 12078598 TI - One-step implant index verification and maxillomandibular relationship. AB - Before definitive fabrication of an implant-supported prosthesis, the accurate positioning of the fixtures or abutment replicas on the working master cast must be verified. In addition, the implant master cast must be accurately mounted against the opposing cast. The purpose of this paper is to demonstrate a technique that combines the verification index and a maxillomandibular relationship into one step, with an accurate technique to record the relationship. PMID- 12078600 TI - Growth of bioactive surfaces on dental implants. AB - Some metallic materials, such as pure titanium, Ti-6Al-4V, are used for dental and orthopedic implants under load-bearing conditions. However, they do not form a chemical bond with bone, which would achieve a good implant-bone fixation in service. In recent works, it has been demonstrated that an in vitro, chemically deposited, bone-like apatite layer with bone-bonding ability could be induced on a titanium surface. By reproducing that chemical procedure, in this work, a dense bone-like apatite layer was formed on the surface of the titanium in simulated body fluid. In addition, the different steps and kinetics of the layer-formation have been studied, because the observation of the samples in the wet state by means of the environmental scanning electron microscope has allowed the observation in situ of the apatite deposition process over a number of days. One of the most important features of the present study is that it can be carried out on a single titanium sample and the process is not interrupted at any stage. One of the main drawbacks of this chemical method is that the samples covered with apatite are susceptible to contamination by bacteria. The behavior of different types of antibiotics used to avoid this contamination has also been studied using the environmental scanning electron microscope. Finally, osteoblast cells have been cultured on the apatite-coated titanium samples to assess their biocompatibility. PMID- 12078601 TI - Implant education: beyond show-and-tell. PMID- 12078599 TI - Crestal bone loss around submerged and exposed unloaded dental implants: a radiographic and microbiological descriptive study. AB - The successful maintenance of crestal bone surrounding dental implants is imperative for long-term implant success. Crestal bone loss is reportedly related to stress. However, early perforation and partial exposure of the implant's covering device are a focus for plaque accumulation, which, if left untreated, may result in inflammation. The objective of this study was to evaluate the crestal bone levels adjacent to submerged and exposed unloaded dental implants during the initial healing phase. In addition, the microbiota around exposed implants were studied. Bilateral implants were placed in the mandible of 10 patients. In one quadrant, the implants were covered by the flap. In the other quadrant, the flap was sutured, leaving the cover screws completely exposed. Standardized periapical radiographs were obtained at implant placement and 4 months later. Radiographs were digitalized, aligned, and analyzed with a computer assisted method. Cultures were obtained from exposed implant sites. All patients showed more crestal bone loss around exposed dental implants compared to submerged implants. Prevotella sp., Streptococcus beta-hemoliticus, and Fusobacterium sp. were the microorganisms identified in most of the sites. The exposure of the implant covering device created foci for bacterial plaque accumulation, which may have facilitated periimplant crestal bone loss. The initial healing phase follow-up may be critical for implant success. PMID- 12078604 TI - [The nurse and night work: living life backwards]. PMID- 12078605 TI - ["My nights are much nicer than your days" Interview by Pierre Luton]. PMID- 12078606 TI - [What is infusion therapy?]. PMID- 12078607 TI - [Infusions. Equipment]. PMID- 12078608 TI - [Preparation, management and monitoring of an infusion]. PMID- 12078610 TI - [It would be better to prevent these meningitides]. PMID- 12078609 TI - [Infusions. Traceability]. PMID- 12078611 TI - [Partners against rheumatoid polyarthritis]. PMID- 12078612 TI - [Children and death]. PMID- 12078613 TI - [Sedentary life style, modern scourge, risk factor]. PMID- 12078614 TI - [Indemnisation for medical errors, (Law no. 2002-303 of March 4, 2002. Title 4- Reparation for consequences of health care risks]. PMID- 12078615 TI - [Bupropion. An antidepressant used to help smoking cessation]. PMID- 12078616 TI - [Insulin injection. Technique]. PMID- 12078617 TI - [Towards a first year of communal education for health professions]. PMID- 12078618 TI - [Home infusion therapy]. PMID- 12078619 TI - [Back pain]. PMID- 12078620 TI - [The public health nurse. A misunderstood profession with multiple missions]. PMID- 12078621 TI - [What do you know about Hirschsprung disease?]. PMID- 12078624 TI - Patterns in midwives' and expectant/new parents' ways of relating to each other in ante- and postnatal consultations. AB - Patterns in midwives' and expectant/new parents' ways of relating to each other in ante- and postnatal midwifery consultations were described by means of qualitative content analysis of 58 video-recorded consultations in a Swedish primary care setting. The midwives appeared to steer the consultations by adopting a basic pattern and occasionally including other patterns. Of the five patterns disclosed, three were revealed as basic and named 'the respectful gardener and her developing plants', 'the propagandist teacher and her ignorant pupils' and 'the steering inspector and the representatives of the population at disposal'. The midwives related in individually oriented ways in the first pattern while the latter two indicated generalized ways of relating. The expectant/new mothers were more active in the two additional patterns 'the mediating counsellor and the discreet seekers of guidance' and 'the personal women-friends'. The expectant/new fathers mostly had a minor role, only appearingly actively involved within the 'gardener' pattern. Midwives need to reflect over their ways of relating as these influence how expectant/new parents deal with the information given and their satisfaction with the care. PMID- 12078625 TI - Caring dynamics as perceived by staff supporting daily occupations for developmentally disabled adults. AB - This study addresses caring staff experiences of hindrances and help in the support of daily occupations among people with developmental disabilities. Data were collected by means of a questionnaire consisting of open-ended questions about the staff perceptions of their work experiences. The respondents (n = 81), corresponding to 94.1% of all care staff employed in a geographically defined care area in southern Sweden, worked in various day activity units supporting the daily occupations of their clients. A constant comparative method of data analysis was used. Staff expressions were classified in two main categories of caring dynamics: an operational level and a managerial level. Four areas were identified at the operational level: encountering realities of practice, attitudes to the clients and work demand, using the potential of knowledge and strategies and applying helping actions to the client. The managerial level included two areas, generalized work strategy and individualized work strategy. It is suggested that to develop the quality of interventions for supporting daily activities among persons with developmental disabilities, efforts should be made to identify caring dynamics as experienced by the caring staff. PMID- 12078626 TI - Auditing nursing content in patient records. AB - The objective of this paper is twofold: (a) to explore different approaches in reviewing records based on a literature review of studies of audits of patient records and (b) to apply these approaches on a sample of records to illuminate consequences of their application. The method used was a literature review of papers on recording of nursing care (n = 56). Based on our findings, an audit of a stratified sample of records (n = 298) from Swedish community health care and nursing homes was performed, applying the different approaches for auditing previously described in the literature. The review showed that audits of patient records were performed using four different approaches with varying aims. The focus of the four approaches can be described as formal structure, process comprehensiveness, knowledge-based and concordance with actual care. The results of this study suggest that audits of patient records should not be solely limited to encompass the formal structure of recording. To avoid a superficial picture or a false sense of high quality and to obtain a more complete and reliable portrait of the quality of recording, we suggest the process comprehensiveness approach in combination with a critical review of the knowledge base for the assessment, diagnosis and interventions of patient records. PMID- 12078627 TI - Interaction between adult patients' family members and nursing staff on a hospital ward. AB - The purpose of this study was to generate knowledge of the interaction between an adult patient's family members and nursing staff from the staff's perspective. Data were collected from nursing staff (n = 155) working on the wards and out patient departments for pulmonary, rheumatic, neurological and gastroenterological diseases at a university hospital by using a new questionnaire based on earlier research and the literature. The questions explored the staff's views of interaction with the adult patient's family members. In this study, interaction is seen as an umbrella concept which encompasses giving information to relatives, discussion, contacts between staff and significant others and working together. The instrument included questions about personal and telephone discussions, the provision of written instructions and factors facilitating and complicating interaction. The response rate was 55%. The data were analysed using SPSS software and examined using frequency and percentage distributions and cross-tabulation. The open-ended questions were analysed using qualitative content analysis by reducing, grouping and abstracting the data inductively. Discussions with relatives while they visited the patient in hospital were the commonest form of interaction. The majority of respondents perceived the interaction with the patient and knowing his or her family members as important. Less than one-fourth of the respondents started discussion with family members, while the majority expected family members to initiate interaction. The majority of respondents perceived the patient's presence in discussion as important, but sometimes they thought it was necessary to discuss with family members without the patient. The staff discussed with family members mainly in the ward office or in patient rooms, which were, however, not perceived as peaceful. Discussions primarily pertained to the patient's condition, discharge from hospital and planning of continued treatment. The majority of respondents reported that the opportunity for discussion offered by staff, the patient's positive attitude towards discussion with the family and family members' own interest facilitated interaction. The staff's haste and shift-work, family members' shyness of approaching the staff and the absence of a peaceful place for discussion serve to complicate interaction. Family members are primarily seen as informants and recipients of information. The staff's ability to recognize the importance of family members to successful patient care is relatively adequate. However, insufficient attention is paid to family members and their need for support. PMID- 12078628 TI - Conditions for rehabilitation of older patients with dementia and hip fracture- the perspective of their next of kin. AB - The aim of this study was to describe the conditions for rehabilitation of older patients with dementia and hip fracture from the perspective of their next of kin. Twenty patients at orthopaedic wards were examined postfracture using a short cognitive test. The same number of next of kin answered four open-ended questions about rehabilitation as well as about the patient's physical function. Qualitative content analysis was used to identify categories. The findings indicate that the conditions for rehabilitation of older patients with dementia and hip fracture are related to the patients' competence, specific needs of support in the light of competence, environmental factors and classification of the rehabilitation activities. The study confirms that the conditions for rehabilitation are related to symptoms of dementia disease and arise from a decline in competence making the patient unable to cope with the environmental pressure and to perform rehabilitation activities. Because of difficulties in assessing competence, patients with dementia are being judged as incapable of managing rehabilitation. A supportive strategy is necessary to encourage the recovery process. PMID- 12078629 TI - The relationship between coping strategies and family burden among relatives of admitted psychiatric patients. AB - Coping strategies as assessed by a projective coping instrument, showed only a weak relationship to perceived family burden and participation in care among 78 close relatives of voluntarily and compulsorily admitted psychiatric patients in this study conducted in two psychiatric services in Sweden in 1997. The results rather indicate that problem solving coping strategies are used when the relatives are in situations amenable to change and that emotion-focused coping strategies are used in situations that are chronic and unchangeable. Furthermore, no differences in coping strategies were found between the relatives and a Swedish normative sample. No relationships were found between coping strategies and when the relative thought that the patient's mental health had led to mental problems in the relative or if the relative experienced that the relationship with the patient had been negatively affected by the mental illness. The usefulness of coping instruments based on personality components in research on family burden is called into question, and a reconsideration of qualitative research methods is recommended. PMID- 12078630 TI - The use of music to decrease agitated behaviour of the demented elderly: the state of the science. AB - This paper reviews the state of the science of interventions using music to decrease the agitated behaviour of the demented elderly person. Seven research articles were located through computerized databases. The review of the literature suggested that music therapy is a useful intervention to help patients deal with a range of behaviour problems. However, overall weakness and limitations of studies are considerable. More rigorous research designs are required to evaluate the immediate and sustained physiological, psychological and sociological effects of music therapy on agitation behaviours of demented elderly. Some recommendations for future research are provided. PMID- 12078631 TI - Congruence between health professionals' and patients' pain ratings: a review of the literature. AB - The literature suggests that health professionals have a tendency to underestimate pain when performing clinical assessments. In addition, it appears that the more clinical experience one has, the greater will be the underestimation of pain. Pain assessment is difficult because of the complex interaction between environment, patient and practitioner variables. Although there is a need for further research in this area, there are clinical implications worthy of consideration. Health professionals need to use a variety of valid and reliable measures, use measures of pain behaviour and disability to complement self-report measures and examine how their own biases and values may influence pain judgements. PMID- 12078632 TI - Physical and psychological health and social relations in patients with prolonged musculoskeletal disorders. AB - The purpose of this study was to describe and analyse the relationship between physical and psychological health and social relations in patients with prolonged musculoskeletal disorders in primary care. Seventy-eight patients--64 women and 14 men--were included in the study. Data from a semi-structured interview called addiction severity index (ASI), frequently used in psychiatric and drug abuse research and a visual analogue scale (VAS) symptom list as well were analysed using principal component analysis (PCA). The analysis gave a two-dimensional model with four poles of related variables characterizing the whole group. The first dimension consisted of one pole characterized by present psychiatric symptoms and many somatic symptoms. The second pole on this dimension was characterized by well-being, having work and feeling satisfaction with life. One pole on the second dimension was characterized by severe physical problems, related to sick leave and disability pension. The opposite pole on this dimension was characterized by severe psychiatric symptoms earlier in life and present relationship problems with family and other people. The ASI-instrument was considered to be relevant for describing important problem areas in patients with prolonged musculoskeletal disorders. PMID- 12078633 TI - The applicability of the Antonovsky Sense of Coherence Scale to a group of Pentecostalists. AB - A methodological issue concerning the Antonovsky of Sense of Coherence (SOC) scale was raised in an earlier study questioning the applicability of the scale in different populations, and among these the Pentecostalists. The question was raised when a group of Pentecostalists had had difficulties in filling in another scale intended to measure the SOC. Therefore, the aim of this study was to test the applicability of the 29-item Antonovsky SOC scale in a group of people (n = 37) belonging to a church of the Pentecostal Movement and to compare the results with those of a randomized group (n = 145) from a general population. The Pentecostalists filled in the scale with no obvious difficulties. No significant differences were found between the Pentecostal group and the group of a general population regarding the total SOC scale scores (mean 152, SD 16 and mean 151, SD 18, respectively). The applicability difficulties found in the earlier study, however using an other SOC scale, could not be confirmed in the present study. To conclude, the Antonovsky SOC scale has so far not shown to have applicability weakness. PMID- 12078634 TI - Towards resolving the interordinal relationships of placental mammals. PMID- 12078635 TI - The fossil record of North American mammals: evidence for a Paleocene evolutionary radiation. AB - Paleontologists long have argued that the most important evolutionary radiation of mammals occurred during the early Cenozoic, if not that all eutherians originated from a single common post-Cretaceous ancestor. Nonetheless, several recent molecular analyses claim to show that because several interordinal splits occurred during the Cretaceous, a major therian radiation was then underway. This claim conflicts with statistical evidence from the well-sampled latest Cretaceous and Cenozoic North American fossil record. Paleofaunal data confirm that there were fewer mammalian species during the latest Cretaceous than during any interval of the Cenozoic, and that a massive diversification took place during the early Paleocene, immediately after a mass extinction. Measurement data show that Cretaceous mammals were on average small and occupied a narrow range of body sizes; after the Cretaceous-Tertiary mass extinction, there was a rapid and permanent shift in the mean. The fact that there was an early Cenozoic mammalian radiation is entirely compatible with the existence of a few Cretaceous splits among modern mammal lineages. PMID- 12078636 TI - Assessing the Cretaceous superordinal divergence times within birds and placental mammals by using whole mitochondrial protein sequences and an extended statistical framework. AB - Using the set of all vertebrate mtDNA protein sequences published as of May 1998, plus unpublished examples for elephant and birds, we examined divergence times in Placentalia and Aves. Using a parsimony-based test, we identified a subset of slower evolutionary rate placental sequences that do not appear to violate the clock assumption. Analyzing just these sequences decreases support for Marsupionta and the carnivore + perissodactyl group but increases support for armadillo diverging earlier than rabbit (which may represent the whole Glires group). A major theme of the paper is to use more comprehensive estimates of divergence time standard error (SE). From the well-studied horse/rhino split, estimated to be 55 million years before present (mybp), the splitting time within carnivores is confidently shown to be older than 50 million years. Some of our estimates of divergence times within placentals are relatively old, at up to 169 million years, but are within 2 SE of other published estimates. The whale/cow split at 65 mybp may be older than commonly assumed. All the sampled splits between the main groups of fereuungulates (the clade of carnivores, cetartiodactyls, perissodactyls, and pholidotes) seem to be distinctly before the Cretaceous/Tertiary boundary. Analyses suggest a close relationship between elephants (representing Afrotheria) and armadillos (Xenarthra), and our timing of this splitting is coincident with the opening of the South Atlantic, a major vicariant event. Recalibrating with this event (at 100 mybp), we obtain younger estimates for the earliest splits among placentals. Divergence times within birds are also assessed by using previously unpublished sequences. We fail to reject a clock for all bird taxa available. Unfortunately, available deep calibration points for birds are questionable, so a new calibration based on the age of the Anseriform stem lineage is estimated. The divergence time of rhea and ostrich may be much more recent than commonly assumed, while that of passerines may be older. Our major concern is the rooting point of the bird subtree, as the nearest outgroup (alligator) is very distant. PMID- 12078637 TI - Interordinal relationships of birds and other reptiles based on whole mitochondrial genomes. AB - Several different groups of birds have been proposed as being the oldest or earliest diverging extant lineage within the avian phylogenetic tree, particularly ratites (Struthioniformes), waterfowl (Anseriformes), and shorebirds (Charadriiformes). Difficulty in resolving this issue stems from several factors, including the relatively rapid radiation of primary (ordinal) bird lineages and the lack of characters from an extant outgroup for birds that is closely related to them by measure of time. To help resolve this question, we have sequenced entire mitochondrial genomes for five birds (a rhea, a duck, a falcon, and two perching birds), one crocodilian, and one turtle. Maximum parsimony and maximum likelihood analyses of these new sequences together with published sequences (18 taxa total) yield the same optimal tree topology, in which a perching bird (Passeriformes) is sister to all the other bird taxa. A basal position for waterfowl among the bird study taxa is rejected by maximum likelihood analyses. However, neither the conventional view, in which ratites (including rhea) are basal to other birds, nor tree topologies with falcon or chicken basal among birds could be rejected in the same manner. In likelihood analyses of a subset of seven birds, alligator, and turtle (9 taxa total), we find that increasing the number of parameters in the model shifts the optimal topology from one with a perching bird basal among birds to the conventional view with ratites diverging basally; moreover, likelihood scores for the two trees are not significantly different. Thus, although our largest set of taxa and characters supports a tree with perching birds diverging basally among birds, the position of this earliest divergence among birds appears unstable. Our analyses indicate a sister relationship between a waterfowl/chicken clade and ratites, relative to perching birds and falcon. We find support for a sister relationship between turtles and a bird/crocodilian clade, and for rejecting both the Haemothermia hypothesis (birds and mammals as sister taxa) and the placement of turtles as basal within the phylogenetic tree for amniote animals. PMID- 12078638 TI - Misinformative characters and phylogeny shape. AB - The discrepancy between theoretical and observed distributions of tree shapes in recent surveys of phylogeny estimates has lead to investigations of possible biological and methodological causes. I investigated whether the phylogenetic quality of characters is related to the tree shape on which they evolve. Simulated evolution revealed shape-related tendencies for characters to indicate correct cladistic relationships; these differences were measured by examining the characters directly, without deriving any phylogeny estimates. Tree stemminess indices correlated strongly with character quality when characters evolved either speciationally or phyletically. Tree balance was a significant correlate of character quality under speciational evolution but not under phyletic evolution. These results help explain the findings of other simulation studies. With additional study of the behavior of evolving characters and their interaction with phylogenetic methods, we might be able to increase the accuracy of tree estimation and compensate for potential biases related to shape. These results give further reason for caution in trusting phylogeny estimates. PMID- 12078639 TI - Power of the concentrated changes test for correlated evolution. AB - The concentrated changes test (CCT) calculates the probability that changes in a binary character are distributed randomly on the branches of a cladogram. This test is used to examine hypotheses of correlated evolution, especially cases where changes in the state of one character influence changes in the state of another character. The test may be sensitive to the addition of branches that lack either trait of interest (white branches). To examine the effects of the proportion of white branches and of tree topology on the CCT probability, we conducted a simulation analysis using a series of randomly generated 100-taxon trees, in addition to a nearly perfectly balanced (symmetrical) and a completely imbalanced (asymmetrical) 100-taxon tree. Using two models of evolution (gains only, or gains and losses), we evolved character pairs randomly onto these trees to simulate cases where (1) characters evolve independently (i.e., no correlation among the traits) or (2) all changes in the dependent character occur on branches containing the independent trait (i.e., a strong correlation among the traits). This allowed us to evaluate the sensitivity of the CCT to type I and type II errors, respectively. In the simulations, the CCT did not appear to be overly sensitive to the inclusion of white branches (low likelihood of type I error with both CCT probabilities < 0.05 and < 0.01). However, the CCT was susceptible to type II error when the proportion of white branches was < 20%. The test was also sensitive to tree shape and was positively correlated to Colless's tree imbalance statistic I. Finally, the CCT responded differently for simulations where only gains were allowed and those where both gains and losses were permitted. These results indicate that the CCT is unlikely to detect a correlation between characters when no such correlation exists. However, when a trait can be gained but not lost, the CCT is conservative and may fail to detect true correlations among traits (increased type II error). Determination of the sampling universe (the taxa included in the comparative analysis) can strongly influence the probability of making such type II errors. We suggest guidelines to circumvent these limitations. PMID- 12078640 TI - Multivariate regression models and geometric morphometrics: the search for causal factors in the analysis of shape. PMID- 12078641 TI - Delimitation of phylogenetic species with DNA sequences: a critique of Davis and Nixon's population aggregation analysis. PMID- 12078642 TI - Ankle morphology of the earliest Cetaceans and its implications for the phylogenetic relations among ungulates. AB - Recent molecular studies are inconsistent with ungulate phylogenetic trees that are based on morphological traits. These inconsistencies especially relate to the position of cetaceans and perissodactyls. Evaluation of the close phylogenetic ties between artiodactyls and cetaceans has been hampered by the absence of tarsal bones of primitive cetaceans, as artiodactyls are often diagnosed on the basis of their tarsus. We here describe newly discovered tarsal bones that are the oldest cetacean tarsals known. We present a character analysis for primitive ungulate tarsals and evaluate their impact on the ungulate phylogenetic tree. Tarsal data are consistent with some molecular studies in suggesting that the extant sister group of Cetacea is Artiodactyla or that Cetacea should be included within the latter order. Tarsal data do not support Cete (Mesonychia plus Cetacea) and are consistent with the exclusion of perissodactyls from paenungulates as suggested by some molecular studies. PMID- 12078643 TI - Using novel phylogenetic methods to evaluate mammalian mtDNA, including amino acid-invariant sites-LogDet plus site stripping, to detect internal conflicts in the data, with special reference to the positions of hedgehog, armadillo, and elephant. AB - We look at the higher-order phylogeny of mammals, analyzing in detail the complete mtDNA sequences of more than 40 species. We test the support for several proposed superordinal relationships. To this end, we apply a number of recently programmed methods and approaches, plus better-established methods. New pairwise tests show highly significant evidence that amino acid frequencies are changing among nearly all the genomes studied when unvaried sites are ignored. LogDet amino acid distances, with modifications to take into account invariant sites, are combined with bootstrapping and the Neighbor Joining algorithm to account for these violations of standard models. To weight the more slowly evolving sites, we exclude the more rapidly evolving sites from the data by using "site stripping". This leads to changing optimal trees with nearly all methods. The bootstrap support for many hypotheses varies widely between methods, and few hypotheses can claim unanimous support from these data. Rather, we uncover good evidence that many of the earlier branching patterns in the placental subtree could be incorrect, including the placement of the root. The tRNA genes, for example, favor a split between the group hedgehog, rodents, and primates versus all other sequenced placentals. Such a grouping is not ruled out by the amino acid sequence data. A grouping of all rodents plus rabbit, the old Glires hypothesis, is also feasible with stripped amino acid data, and rodent monophyly is also common. The elephant sequence allows confident rejection of the older taxon Ferungulata (Simpson, 1945). In its place, the new taxa Scrotifera and Fereuungulata are defined. A new likelihood ratio test is used to detect differences between the optimal tree for tRNA versus that for amino acids. While not clearly significant as made, some results indicate the test is tending towards significance with more general models of evolution. Individual placement tests suggest alternative positions for hedgehog and elephant. Congruence arguments to support elephant and armadillo together are striking, suggesting a superordinal group composed of Xenarthra and African endemic mammals, which in turn may be near the root of the placental subtree. Thus, while casting doubt on some recent conclusions, the analyses are also unveiling some interesting new possibilities. PMID- 12078644 TI - Phylogenetic assessment of molecular and morphological data for eutherian mammals. AB - The interordinal relationships of eutherian (placental) mammals were evaluated by a phylogenetic analysis of four published data sets (three sequences and one morphological). The nature and degree of support and conflict for particular groups were assessed by separate bootstrap and homogeneity tests, which were followed by combined analyses of the sequence and morphological data. Between orders, strong support (i.e., > or = 95% bootstrap scores) was found for a paraphyletic Artiodactyla (relative to Cetacea) and a monophyletic Cetartiodactyla (Artiodactyla and Cetacea) and Paenungulata (Hyracoidea, Proboscidea, and Sirenia). In turn, some reasonable to strong evidence (> or = 85%) was obtained for Hyracoidea with Sirenia, Dermoptera with Scandentia, Glires (Lagomorpha with Rodentia), and Afrotheria (Amblysomus, Macroscelidea, Paenungulata, and Tubulidentata). Otherwise, no other interordinal clades were supported at these reasonable to strong levels. This overall lack of resolution for eutherian interordinal clusters agrees with other studies that suggest further progress will continue to be slow and difficult. Further resolution will require the integration of more recently published data, the continued sampling of taxa and characters, and the use of more powerful methods of data analysis. PMID- 12078645 TI - Stability of cladistic relationships between Cetacea and higher-level artiodactyl taxa. AB - Over the past 10 years, the phylogenetic relationships among higher-level artiodactyl taxa have been examined with multiple data sets. Many of these data sets suggest that Artiodactyla (even-toed ungulates) is paraphyletic and that Cetacea (whales) represents a highly derived "artiodactyl" subgroup. In this report, phylogenetic relationships between Cetacea and artiodactyls are tested with a combination of 15 published data sets plus new DNA sequence data from two nuclear loci, interphotoreceptor retinoid-binding protein (IRBP) and von Willebrand factor (vWF). The addition of the IRBP and vWF character sets disrupts none of the relationships supported by recent cladistic analyses of the other 15 data sets. Simultaneous analyses support three critical clades: (Cetacea + Hippopotamidae), (Cetacea + Hippopotamidae + Ruminantia), and (Cetacea + Hippopotamidae + Ruminantia + Suina). Perturbations of the combined matrix show that the above clades are stable to a variety of disturbances. A chronicle of phylogenetic results over the past 3 years suggests that cladistic relationships between Cetacea and artiodactyls have been stable to increased taxonomic sampling and to the addition of more than 1,400 informative characters from 15 data sets. PMID- 12078646 TI - Additional support for Afrotheria and Paenungulata, the performance of mitochondrial versus nuclear genes, and the impact of data partitions with heterogeneous base composition. AB - We concatenated sequences for four mitochondrial genes (12S rRNA, tRNA valine, 16S rRNA, cytochrome b) and four nuclear genes [aquaporin, alpha 2B adrenergic receptor (A2AB), interphotoreceptor retinoid-binding protein (IRBP), von Willebrand factor (vWF)] into a multigene data set representing 11 eutherian orders (Artiodactyla, Hyracoidea, Insectivora, Lagomorpha, Macroscelidea, Perissodactyla, Primates, Proboscidea, Rodentia, Sirenia, Tubulidentata). Within this data set, we recognized nine mitochondrial partitions (both stems and loops, for each of 12S rRNA, tRNA valine, and 16S rRNA; and first, second, and third codon positions of cytochrome b) and 12 nuclear partitions (first, second, and third codon positions, respectively, of each of the four nuclear genes). Four of the 21 partitions (third positions of cytochrome b, A2AB, IRBP, and vWF) showed significant heterogeneity in base composition across taxa. Phylogenetic analyses (parsimony, minimum evolution, maximum likelihood) based on sequences for all 21 partitions provide 99-100% bootstrap support for Afrotheria and Paenungulata. With the elimination of the four partitions exhibiting heterogeneity in base composition, there is also high bootstrap support (89-100%) for cow + horse. Statistical tests reject Altungulata, Anagalida, and Ungulata. Data set heterogeneity between mitochondrial and nuclear genes is most evident when all partitions are included in the phylogenetic analyses. Mitochondrial-gene trees associate cow with horse, whereas nuclear-gene trees associate cow with hedgehog and these two with horse. However, after eliminating third positions of A2AB, IRBP, and vWF, nuclear data agree with mitochondrial data in supporting cow + horse. Nuclear genes provide stronger support for both Afrotheria and Paenungulata. Removal of third positions of cytochrome b results in improved performance for the mitochondrial genes in recovering these clades. PMID- 12078647 TI - Mammalian evolution: timing and implications from using the LogDeterminant transform for proteins of differing amino acid composition. AB - We explore the tree of mammalian mtDNA sequences, using particularly the LogDet transform on amino acid sequences, the distance Hadamard transform, and the Closest Tree selection criterion. The amino acid composition of different species show significant differences, even within mammals. After compensating for these differences, nearest-neighbor bootstrap results suggest that the tree is locally stable, though a few groups show slightly greater rearrangements when a large proportion of the constant sites are removed. Many parts of the trees we obtain agree with those on published protein ML trees. Interesting results include a preference for rodent monophyly. The detection of a few alternative signals to those on the optimal tree were obtained using the distance Hadamard transform (with results expressed as a Lento plot). One rearrangement suggested was the interchange of the position of primates and rodents on the optimal tree. The basic stability of the tree, combined with two calibration points (whale/cow and horse/rhinoceros), together with a distant secondary calibration from the mammal/bird divergence, allows inferences of the times of divergence of putative clades. Allowing for sampling variances due to finite sequence length, most major divergences amongst lineages leading to modern orders, appear to occur well before the Cretaceous/Tertiary (K/T) boundary. Implications arising from these early divergences are discussed, particularly the possibility of competition between the small dinosaurs and the new mammal clades. PMID- 12078648 TI - The virtues of gaps: xenarthran (Edentate) monophyly supported by a unique deletion in alpha A-crystallin. AB - Shared insertions or deletions (indels) in protein-coding DNA can be strong indicators of the monophyly of a taxon. A three-amino acid deletion had previously been noted in the eye lens protein alpha A-crystallin of two species of sloths and two species of anteaters, which represent the Pilosa, one of the two infraorders of Xenarthra (Edentata). This deletion has not been observed in 55 species from 16 other eutherian orders, or in 2 species of marsupials, or in 34 nonmammalian vertebrates, from birds to shark. At the genomic level, we have now detected this deletion in two species of armadillos of the second xenarthran infraorder, Cingulata, as well as in an additional species of anteater. Phylogenetic trees were constructed from a 145-bp sequence of the alpha A crystallin gene of 39 tetrapod species, supporting xenarthran monophyly with values from 76% to 90%. To quantify the additional support for xenarthran monophyly, as given by the three-residue deletion, we computed the probabilities for the occurrence of this deletion per evolutionary time unit for alternative hypothetical tree topologies. In the estimates obtained, the six trees in which the xenarthran subgroups are unresolved or paraphyletic give an increasingly lower likelihood than do the two trees that assume xenarthran monophyly. For the monophyletic trees, the probability that the deletion observed in the xenarthrans is due to a single event is > 0.99. Thus, this deletion in alpha A-crystallin gives strong molecular support for the monophyly of this old and diverse order. PMID- 12078649 TI - [Treatment of attention deficit disorder with hyperactivity]. PMID- 12078650 TI - Identification of HLA-A2 restricted CD8(+) T-lymphocyte responses to Plasmodium vivax circumsporozoite protein in individuals naturally exposed to malaria. AB - Specific CD8(-) T-lymphocyte (CTL) activity against Plasmodium pre-erythrocytic stages (P-ES) derived antigens is considered one of the most important mechanisms for malaria protection. Plasmodium vivax is the second most prevalent human malaria parasite species distributed worldwide. Although several CTL epitopes have been identified in Plasmodium falciparum P-ES derived antigens, none has been described for P. vivax to date. In this study, we analysed HLA-A*0201 specific CD8(-) T-lymphocyte responses to the P. vivax circumsporozoite (CS) protein in both malaria exposed and non-exposed populations from the Colombian Pacific Coast. First, we analysed the prevalence of HLA-A2 allele in the study populations and found that approximately 38 of the individuals expressed this molecule and that 50 of them were HLA-A*0201. We then selected, on the P. vivax CS, five peptide sequences containing the HLA-A*0201 binding motifs and used the corresponding synthetic peptides to evaluate the CD8(-) T-lymphocyte interferon (IFN)-gamma response. Peripheral blood mononuclear cells from the HLA-A*0201 donors were in vitro stimulated with these peptides and IFN-gamma production was determined by an ELISPOT assay. Specific CD8(-) T-lymphocyte responses were detected for three peptides located in the C-terminal region of the protein. Specific responses to these peptides were also detected in several individuals expressing different HLA-A*02 subtypes. The potential of these peptides to induce specific cytolysis and that of long synthetic peptides comprising these epitopes as P. vivax malaria vaccine subunits are being studied. PMID- 12078651 TI - Management of thoracic duct injuries after oesophagectomy (Br J Surg 2001: 88: 1442-8). Letter 2. PMID- 12078652 TI - [Basic concept of management of adult community-acquired pneumonia]. PMID- 12078653 TI - Is the gastrointestinal tract the sole source of organisms in ventilator associated pneumonia? PMID- 12078654 TI - AARC Clinical Practice Guideline: selection of an oxygen delivery device for neonatal and pediatric patients--2002 revision & update. PMID- 12078655 TI - AARC Clinical Practice Guideline: oxygen therapy for adults in the acute care facility--2002 revision & update. PMID- 12078656 TI - AARC Clinical Practice Guideline: in-hospital transport of the mechanically ventilated patient--2002 revision & update. PMID- 12078657 TI - Proceeding of the consensus conference of the role of sentinel lymph node biopsy in carcinoma or the breast April 19-22, 2001, Philadelphia, PA, USA. AB - A consensus conference on the role of sentinel node biopsy in breast cancer was held in Philadelphia in april, 2001; the participants included many highly respected American and European investigators in this area. This report summarizes the deliberations of the group and promotes its current guidelines for the integration of this new technique into contemporary clinical practice. PMID- 12078658 TI - Blue baby syndrome. PMID- 12078660 TI - Bibliography. Current World Literature. Neoplasms of the lungs. PMID- 12078659 TI - Enteral nutritional supplementation and the maintenance of permission in Crohn's disease. PMID- 12078661 TI - Bibliography. Current World Literature. Diseases of the pleura. PMID- 12078662 TI - AGA program at DDW--a success. PMID- 12078663 TI - Gastroenterology research in Melbourne, Australia. PMID- 12078664 TI - The evolution of modern academic medicine continues. PMID- 12078665 TI - Hemobiliary from arterial pseudoaneurysm. PMID- 12078666 TI - The winner after 7 rounds--fecal occult blood testing! PMID- 12078667 TI - Forecast for using metabolite measurements in the dosing of azathioprine or 6 mercaptopurine for IBD patients: "partly cloudy". PMID- 12078668 TI - Persistence and pathogenesis: a great defense facilitates a good offense. PMID- 12078669 TI - Celiac sprue as a possible cause of symptoms in presumed irritable bowel syndrome. PMID- 12078670 TI - Involuntary movements in infantile cobalamin deficiency appearing after treatment. PMID- 12078671 TI - Involuntary movements in infantile cobalamin deficiency appearing after treatment. PMID- 12078672 TI - Multipass reconfiguration of the HELEN Nd:glass laser at the Atomic Weapons Establishment. AB - The HELEN high-power Nd:glass laser has been rebuilt in a new multipass configuration that requires fewer components to maintain existing performance. This is expected to lead to greater system availability and reduced running costs. We describe the new design, discuss some of the key issues that had to be addressed, and present operational results. PMID- 12078673 TI - Micro-electro-mechanical system-based digitally controlled optical beam profiler. AB - An optical beam profiler is introduced that uses a two-dimensional (2-D) small tilt micromirror device. Its key features include fast speed, digital control, low polarization sensitivity, and wavelength independence. The use of this 2-D multipixel device opens up the important possibility of realizing several beam profile measurement concepts, such as a moving knife edge, a scanning slit, a moving pinhole, a variable aperture, and a 2-D photodiode array. The experimental proof of the optical beam profiler concept using a 2-D digital micromirror device to simulate the 2-D moving knife edge indicates a small measurement error of 0.19% compared with the expected number based on a Gaussian beam-propagation analysis. Other 2-D pixel arrays such as a liquid-crystal-based 90 degrees polarization rotator sandwiched between crossed polarizers can also be exploited for the optical beam whose polarization direction is known. PMID- 12078674 TI - Suppression of self-pulsing behavior in erbium-doped fiber lasers with a semiconductor optical amplifier. AB - We experimentally demonstrate that the stability of cw and mode-locked erbium doped fiber ring lasers can be improved significantly with a semiconductor optical amplifier (SOA) inside the cavity. The fast saturable gain of the SOA suppresses significantly the self-pulsing that is due to ion pairs in the erbium doped fiber, which acts as a saturable absorber. A linear stabilization analysis of the laser system agrees with our experimental results. PMID- 12078675 TI - Method of spectral compensation and its application in the ultraviolet for sulfur dioxide control with a gas-filter correlation instrument. AB - Changes in structureless spectra that are due to the presence of interfering absorbers or light source instabilities, changes in the spectral transfer function of the optics, and changes in the detector's spectral responsivity degrade measurement accuracy. A method of compensating for changes in structureless spectra is developed for a gas-filter correlation instrument. It is shown that there are points in the spectrum where the effect of the interfering component's having a structureless spectrum on the measurement can be drastically reduced. PMID- 12078676 TI - Bandpass-resampling effects for the retrieval of surface emissivity. AB - The retrieval of surface emissivity in the 8-14-microm region from remotely sensed thermal imagery requires channel-averaged values of atmospheric transmittance, path radiance, and downwelling sky flux. Band-pass resampling introduces inherent retrieval errors that depend on atmospheric conditions, spectral region, bandwidth, flight altitude, and surface temperature. This simulation study is performed for clear sky conditions and moderate atmospheric water vapor contents. It shows that relative emissivity retrieval errors can reach as much as 3% for broadband sensors (1-2-microm bandwidth) and 0.8% for narrowband instruments (0.15 microm), even for constant surface emissivity. For spectrally varying surface emissivities the relative retrieval error increases for the broadband instrument by approximately 2% in channels with strong emissivity changes of 0.05-0.1. The corresponding retrieval errors for narrowband sensors increase by approximately 3-4%. The channels in the atmospheric window regions with lower transmittance, i.e., 8-8.5 and 12.5-14 microm, are most sensitive to retrieval errors. PMID- 12078677 TI - Radiometric calibration of an airborne CO2 pulsed Doppler lidar with a natural earth surface. AB - Radiometric calibration of an airborne CO2 pulsed Doppler lidar has been accomplished with surface retroreflection signals from the White Sands National Monument, New Mexico. Two circular passes were made at altitudes of 6.3 and 9.3 km. The computed calibration factors for both altitudes are in excellent agreement with the value derived from standard ground-based measurements involving a fixed sandpaper target of known reflectance. This finding corroborates a previous study that successfully calibrated an airborne cw Doppler lidar with a variety of natural Earth surfaces. The present results indicate that relatively uniform Earth surface targets can be used for in-flight calibration of CO2 pulsed airborne and, in principal, other infrared lidars. PMID- 12078678 TI - Concentration measurement of gas embedded in scattering media by employing absorption and time-resolved laser spectroscopy. AB - Diode-laser-based absorption spectroscopy for the evaluation of embedded gas concentrations in porous materials is demonstrated in measurements of molecular oxygen dispersed throughout scattering polystyrene foam, used here as a generic test material. The mean path length of light scattered in the material is determined with the temporal characteristics of the radiation transmitted through the sample. This combined with sensitive gas-absorption measurements employing wavelength-modulation spectroscopy yields an oxygen concentration in polystyrene foam of 20.4% corresponding to a foam porosity of 98%, which is consistent with manufacturing specifications. This feasibility study opens many possibilities for quantitative measurements by using the method of gas-in-scattering-media absorption spectroscopy. PMID- 12078679 TI - Mie scattering by a spherical particle in an absorbing medium. AB - We show that the large-size parameter limit of the scattering efficiency of a spherical particle of relative refractive index m(r) embedded in an absorbing medium is equal to [m(r) - 1[2/]m(r) + 1]2 and not to zero as has been claimed in a recent article [J. Appl. Opt. 40, 1354-1361 (2001)]. PMID- 12078680 TI - Strategies for laser-induced fluorescence detection of nitric oxide in high pressure flames. I. A-X(0,0) excitation. AB - Three different high-pressure flame measurement strategies for NO laser-induced fluorescence (LIF) with A-X(0,0) excitation have been studied previously with computational simulations and experiments in flames up to 15 bars. Interference from O2 LIF is a significant problem in lean flames for NO LIF measurements, and pressure broadening and quenching lead to increased interference with increased pressure. We investigate the NO LIF signal strength, interference by hot molecular oxygen, and temperature dependence of the three previous schemes and for two newly chosen excitation schemes with wavelength-resolved LIF measurements in premixed methane and air flames at pressures between 1 and 60 bars and a range of fuel/air ratios. In slightly lean flames with an equivalence ratio of 0.83 at 60 bars, the contribution of O2 LIF to the NO LIF signal varies between 8% and 29% for the previous schemes. The O2 interference is best suppressed with excitation at 226.03 nm. PMID- 12078681 TI - Diffractive optical isolator made of high-efficiency dielectric gratings only. AB - The working principle of an optical isolator made of two corrugated dielectric gratings is introduced. One grating acts as a polarizer, and the other acts as a quarter-wave plate used in conical incidence converting linearly polarized light into circularly polarized light. Global maxima of diffraction efficiency for surface-corrugated gratings with binary, sinusoidal, and pyramidal ridge shapes with dependence on the material index are identified. Regarding technological feasibility for use in the visible wavelength range, high-frequency gratings with a binary shape were realized. With these gratings, an extinction ratio of more than 40 dB for the polarizer is theoretically possible, and more than 20 dB was experimentally achieved. A good correlation between theoretically calculated efficiencies and birefringences based on rigorous methods and the experimental results is demonstrated. PMID- 12078682 TI - Cavity-ring-down principle for fiber-optic resonators: experimental realization of bending loss and evanescent-field sensing. AB - A novel measurement principle for fiber-optic sensing is presented. Use of a cavity-ring-down scheme enables measurements of minute optical losses in high finesse fiber-optic cavities. The loss may be induced by evanescent-field absorption, fiber bending, fiber degradation, Bragg gratings, or any other effect that might change the fiber transmission or cavity reflector properties. The principle is proved to be rather insensitive to ambient perturbations such as temperature changes. A high-sensitivity measurement of loss due to bending is presented as a proof-of-principle. With a cavity finesse of 627 a sensitivity for induced loss of 108 ppm (4.68 x 10(-4) dB) is achieved. Preliminary measurements of evanescent-field absorption are also discussed. PMID- 12078683 TI - Threshold of stimulated Brillouin scattering by use of a solar pumped laser. AB - What is to our knowledge the first stimulated Brillouin scattering experiment using a high-power low-gain solar pumped laser is presented. A threshold reflectivity of 0.23% was reached when a peak power of 20.7 kW was used at 7.6 GHz. A cw solar pumped laser was Q-switched with an acousto-optic modulator, and the bandwidth was narrowed with an intracavity etalon. A high polarization ratio (>99.4%) was achieved by use of an intracavity configuration. Higher reflectivity values were limited because of the lack of availability of optical switches. PMID- 12078684 TI - Absolute solar transmittance interferometer for ground-based measurements. AB - The absolute solar transmittance interferometer measures absolute solar radiance at the Earth's surface. The instrument is based on a Fourier-transform spectrometer that utilizes a liquid-nitrogen-cooled InSb detector and appropriate optical bandpass filters. The recorded solar spectra are calibrated against National Institute of Standards and Technology traceable lamps and a blackbody source. The spectral range addressed by this instrument is from 1950 to 10100 cm( 1) at a resolution of 2 cm(-1). The optical design of the instrument and the experimental methods are discussed. A discussion of the uncertainties involving the instrument and the calibration sources is presented. Initial measurements from several sites are compared with atmospheric model calculations. PMID- 12078685 TI - Superimposed noninterfering probes to extend the capabilities of phase Doppler anemometry. AB - We propose using multiple superimposed noninterfering probes (SNIPs) of the same wavelength but different beam angles to extend the capabilities of phase Doppler anemometry. When a particle is moving in a SNIP the Doppler signals that are produced exhibit multiple Doppler frequencies and phase shifts. The resolution of the measurements of particle size (i.e., by fringe spacing and Doppler frequency) increases with beam angle. Then, with the solution proposed, even with only two detectors several measurements of size can be obtained for the same particle with increasing resolution if we consider higher frequencies in the signal. Several optical solutions to produce SNIPs as well as a signal-processing algorithm to treat the multiple-frequency Doppler signals are proposed. Experimental validations of the sizing of spherical and cylindrical particles demonstrate the applicability of this technique for particle measurement. We believe that this new technique can be of great interest when high resolution of size, velocity, and even refractive index is required. PMID- 12078686 TI - Thermo-optic design for microwave and millimeter-wave electromagnetic power microsensors. AB - Rendina et al. recently proposed the original configuration of an electromagnetic power sensor for microwaves and millimeter waves that is based on an optically interrogated all-silicon chip [Electron. Lett. 35, 1748 (1999)]. Here we theoretically analyze and discuss in detail the performances of such a new class of nonperturbing and wideband probe in terms of sensitivity, resolution, intrinsic detectivity, linearity, and response time. Good agreement between theory and experiments is demonstrated. In particular, minimum resolutions of approximately 1 mW/cm2 are obtained at frequencies beyond 10 GHz. The dependence of response on the geometrical and electromagnetic parameters of the sensing element is analyzed, and on this basis the possibility of achieving optimized configurations is discussed. PMID- 12078687 TI - Tunable diode laser absorption spectroscopy in coiled hollow optical waveguides. AB - We demonstrate tunable diode laser absorption spectroscopy of CO2 and NH3 near 1.5 microm using a distributed feedback diode laser in conjunction with hollow optical waveguides as long-path sample cells. The waveguides are coiled to reduce the physical extent of the system. The small volume of the waveguide provides rapid instrument response to changes in gas concentration. To reduce the pressure drop associated with long lengths and high flow rates, we perforate the waveguides in a novel geometry providing parallel pneumatic paths while maintaining optical path length. A minimum detectable absorbance of 3.5 x 10(-5) in a 3-m section of waveguide is demonstrated. PMID- 12078688 TI - Mid-infrared diffused planar waveguides made of silver halide chloro-bromide. AB - We have developed waveguides for the mid-IR spectrum using silver halide crystals. Diffused planar waveguides constructed from silver chlorobromide (AgClBr) with different diffusion thicknesses were designed and characterized. The waveguides were based on our changing the refractive index of the guiding layer by a diffusion of Br- ions into crystalline AgCl substrates. The waveguides were analyzed and investigated with a 10.6-microm CO2 laser, and the optically measured thicknesses of the waveguides were found to vary between 65 and 600 microm. The propagation losses were in the range of 4-16 dB/cm, and the maximal entrance angle was 62 degrees. The output beam distribution was measured and was in good correlation with a numerical analysis simulation based on a ray-tracing model, by use of the eikonal equation. PMID- 12078689 TI - Investigation of processes leading to damage growth in optical materials for large-aperture lasers. AB - Damage growth in optical materials used in large-aperture laser systems is an issue of great importance to determine component lifetime and therefore cost of operation. Small size damage sites tend to grow when exposed to subsequent high power laser irradiation at 355 nm. An understanding of the photophysical processes associated with damage growth is important to devise mitigation techniques. We examine the role of laser-modified material and cracks formed in the crater of damage pits in the damage growth process using fused-silica and deuterated KDP samples. Experimental results indicate that both of the above mentioned features can initiate plasma formation at fluences as low as 2 J/cm2. The intensity of the recorded plasma emission remains low for fluences up to approximately 5 J/cm2 but rapidly increases thereafter, accompanied by an increase of the size of the damage crater. PMID- 12078690 TI - Laser operation with two orthogonally polarized transverse modes. AB - Laser resonator configurations, which enable laser operation with two orthogonally polarized transverse modes, are presented. The intensity distributions of these two modes can be chosen to be complementary, so the gain medium can be exploited more efficiently than with a single mode, leading to improved output power. Moreover, the two modes can be combined and efficiently transformed into a single high-quality beam. Basic principles and experimental results with Nd:YAG lasers are presented. PMID- 12078691 TI - Bistatic receiver model for airborne lidar returns incident on an imaging array from underwater objects. AB - We develop a bistatic model for airborne lidar returns collected by an imaging array from underwater objects, incorporating additional returns from the surrounding water medium and ocean bottom. Our results provide a generalization of the monostatic model by Walker and McLean. In the bistatic scheme the transmitter and receiver are spatially separated or are not coaligned. This generality is necessary for a precise description of an imaging array such as a CCD, which may be viewed as a collection of receiver elements, with each transmitter-element pair forming a bistatic configuration. More generally, the receiver may consist of photomultiplier tubes, photodiodes, or any of a variety of optical receivers, and the imaging array can range in size from a CCD array to a multiple-platform airborne lidar system involving multiple aircraft. The majority of this research is devoted to a derivation of the bistatic lidar equations, which account for multiple scattering and absorption in the water column. We then describe the application of these equations to the modeling and simulation of an imaging array. We show an example of a simulated lidar return and compare it with a real ocean lidar return, obtained by a CCD array. PMID- 12078692 TI - Determination of the droplet effective size and optical depth of cloudy media from polarimetric measurements: theory. AB - We present the development of a semi-analytical algorithm for optical particle sizing in disperse media. The algorithm is applied to the specific case of water clouds. However, it can be extended with minor modifications to other types of light-scattering medium. It is assumed that the optical thickness tau of the medium is large and the probability of photon absorption beta is small. Thus the optical particle-sizing problem is studied in the regime of highly developed multiple light scattering. It was found that the degree of polarization in visible and near-infrared channels provides us with information both on the effective size of droplets and on the optical thickness tau. PMID- 12078693 TI - Sum-frequency-generation system for differential absorption lidar measurement of atmospheric nitrogen dioxide. AB - A sum-frequency-generation system for differential absorption lidar measurement of atmospheric nitrogen dioxide in the lower troposphere was developed. The system uses a combination of a pair of KD*P crystals and a tunable dye laser with LDS 765 dye pumped by the second harmonic of a Nd:YAG laser to generate lambdaon and lambdaoff alternatively. Compared with the conventional system that uses Coumarin 445 dye pumped by the third harmonic, the output energy and long-term stability were improved. By use of this system, atmospheric NO2 concentrations of approximately 10-50 ppb were measured, with an instrumental error of approximately 7 ppb. PMID- 12078694 TI - Microwindow selection for high-spectral-resolution sounders. AB - The recent development of satellite instruments that obtain spectrally resolved measurements of the atmosphere has highlighted the problem of how to determine the best subsets, or microwindows, of such spectra for retrievals of temperature and composition. A technique is described that maximizes the information content (or some other figure of merit) based on the modeling of the propagation of systematic as well as random error terms through the retrieval process. Apart from selecting microwindows, this technique can also prioritize existing microwindows for different circumstances and provides a full error analysis of the retrieval. A practical application is demonstrated for the Michelson Interferometer for Passive Atmospheric Sounding limb-viewing interferometer, but the technique is equally applicable to nadir-viewing instruments. PMID- 12078695 TI - Evaluation of the performance of a shearing interferometer in strong scintillation in the absence of additive measurement noise. AB - A simulation study is presented that evaluates the ability of a unit-shear, shearing interferometer to estimate a complex field resulting from propagation through extended turbulence. Performance is defined in terms of the Strehl ratio achieved when the estimate of the complex field obtained from reconstruction is used to correct the distorted wave front presented to the wave-front sensor. A series of evaluations is performed to identify the strengths and weaknesses of the shearing interferometer in the two-dimensional space of the Fried parameter r0 and the Rytov number. The performance of the shearing interferometer is compared with that of a Hartmann sensor in the Fried and Hutchin geometries. Although the effects of additive measurement noise (such as read noise, shot noise, amplifier noise) are neglected, the fundamental characteristics of the measurement process are shown to distinguish the performance of the various wave front sensors. It is found that the performance of a shearing interferometer is superior to that of a Hartmann sensor when the Rytov number exceeds 0.2. PMID- 12078696 TI - Inversion with regularization for the retrieval of tropospheric aerosol parameters from multiwavelength lidar sounding. AB - We present an inversion algorithm for the retrieval of particle size distribution parameters, i.e., mean (effective) radius, number, surface area, and volume concentration, and complex refractive index from multiwavelength lidar data. In contrast to the classical Tikhonov method, which accepts only that solution for which the discrepancy reaches its global minimum, in our algorithm we perform the averaging of solutions in the vicinity of this minimum. This averaging stabilizes the underlying ill-posed inverse problem, particularly with respect to the retrieval of number concentration. Results show that, for typical tropospheric particles and 10% error in the optical data, the mean radius could be retrieved to better than 20% from a lidar on the basis of a Nd:YAG laser, which provides a combination of backscatter coefficients at 355, 532, and 1064 nm and extinction coefficients at 355 and 532 nm. The accuracy is improved if the lidar is also equipped with a hydrogen Raman shifter. In this case two additional backscatter coefficients at 416 and 683 nm are available. The combination of two extinction coefficients and five backscatter coefficients then allows one to retrieve not only averaged aerosol parameters but also the size distribution function. There was acceptable agreement between physical particle properties obtained from the evaluation of multiwavelength lidar data taken during the Lindenberg Aerosol Characterization Experiment in 1998 (LACE 98) and in situ data, which were taken aboard aircraft. PMID- 12078697 TI - Heating of three-layer solid aerosol particles by laser radiation. AB - The heating of lidar-irradiated multilayer particles is analyzed theoretically and numerically by solution of the heat conduction equation. The internal intensity and temperature distributions are presented for particles composed of air, quartz, and carbon. It is shown that the heating times of such particles substantially depend on particle radii, layer position, and shell thickness. In particular, the decrease in thickness of the surface carbon layer can result in a reduction of the heating time of multilayer particles. PMID- 12078698 TI - Low-altitude infrared propagation in a coastal zone: refraction and scattering. AB - Midwave and long-wave infrared propagation were measured in the marine atmosphere close to the surface of the ocean. Data were collected near San Diego Bay for two weeks in November 1996 over a 15-km horizontal path. The data are interpreted in terms of effects expected from molecules, aerosol particles, and refraction. Aerosol particles are a dominant influence in this coastal zone. They induce a diurnal variation in transmission as their character changes with regular changes in wind direction. A refractive propagation factor calculation is introduced, and it is systematically applied to the model and to the data analysis. It is shown that this refractive propagation factor is a necessary component of a complete near-sea-surface infrared transmission model. PMID- 12078699 TI - Cloud physics lidar: instrument description and initial measurement results. AB - The new Cloud Physics Lidar (CPL) has been built for use on the NASA ER-2 high altitude aircraft. The purpose of the CPL is to provide multiwavelength measurements of cirrus, subvisual cirrus, and aerosols with high temporal and spatial resolution. The CPL utilizes state-of-the-art technology with a high repetition rate, a low-pulse-energy laser, and photon-counting detection. The first deployment for the CPL was the Southern African Regional Science Initiative's 2000 field campaign during August and September 2000. We provide here an overview of the instrument and initial data results to illustrate the measurement capability of the CPL. PMID- 12078700 TI - Generation and characterization of ultrafast white-light continuum in condensed media. AB - The generation of white-light continuum by femtosecond laser pulses in transparent condensed media is investigated comprehensively with 262-, 393-, and 785-nm pump wavelengths. We find that the ratio of the medium's bandgap energy to the photon energy of the incident wavelength determines the amount of anti-Stokes broadening, independently of the pump wavelength and the medium's bandgap. It is also shown that, although the amount of anti-Stokes broadening is greater for the longer pump wavelength, the shorter pump wavelength is more advantageous for generating shorter-wavelength continua in the UV region. In addition, a self induced change in polarization of the white-light continuum that is generated is observed in an isotropic material with a cubic crystal structure, such as CaF2 and LiF. After the investigation of polarization, the frequency chirp of the continuum is characterized by the Kerr-gate method with 70-fs temporal and 10-nm wavelength resolution. PMID- 12078701 TI - Optical Kerr switching technique for the production of a picosecond, multiwavelength CO2 laser pulse. AB - A wavelength-independent method for optical gating, based on the optical Kerr effect, has been demonstrated. Using this method, we produced 100-ps, 10-kW, two wavelength pulses (10.3 and 10.6 microm) with a signal-to-background ratio contrast of 10(5) by slicing a long CO2 pulse. The capability of gating consecutive pulses separated on a picosecond time scale with this method is also shown. PMID- 12078702 TI - Interferometric correlation of infrared femtosecond pulses with two-photon conductivity in a silicon CCD. AB - We present an experimental setup capable of performing a single-shot interferometric correlation of femtosecond pulses using two-photon conductivity in a standard silicon CCD camera. The method is demonstrated with 100-fs pulses at 1.4 microm. PMID- 12078703 TI - Gas temperature measurements in weakly ionized glow discharges with filtered Rayleigh scattering. AB - We report the first gas temperature measurements in plasmas to our knowledge obtained by filtered Rayleigh scattering (FRS). A narrow-linewidth Ti:sapphire laser is used as the illumination source, and a mercury filter provides strong suppression of elastic background. We perform measurements in weakly ionized glow discharges in pure argon and in an argon-plus-1%-nitrogen mixture. Where possible, we verify the FRS technique by comparing filtered measurements with unfiltered measurements. We present point measurements of axial temperature with uncertainties of less than 5%. We use a planar scheme to obtain radial temperature profiles with uncertainties of 10%. PMID- 12078704 TI - Homocysteine and dementia. PMID- 12078705 TI - Leptin-replacement therapy in lipodystrophy. PMID- 12078706 TI - Leptin-replacement therapy in lipodystrophy. PMID- 12078707 TI - Direct-to-consumer marketing. PMID- 12078708 TI - Direct-to-consumer marketing. PMID- 12078709 TI - Direct-to-consumer marketing. PMID- 12078710 TI - Direct-to-consumer marketing. PMID- 12078711 TI - Debt repayment for trainees. PMID- 12078712 TI - Debt repayment for trainees. PMID- 12078713 TI - All you need is cognitive behaviour therapy? Integrated biopsychosocial approach will be treatment of choice. PMID- 12078714 TI - All you need is cognitive behaviour therapy? Political range of psychotherapies has emerged. PMID- 12078715 TI - The limits of psychiatry. People with ADHD are not lazy, unmotivated, and irresponsible. PMID- 12078716 TI - Common pitfalls in bioinformatics-based analyses: look before you leap. PMID- 12078717 TI - Toward an ethical consultation in intensive care? PMID- 12078718 TI - Effects of roughage source and concentration on intake and performance by finishing heifers. AB - Three experiments were conducted to evaluate the effects of roughage source and concentration on intake and performance by finishing heifers. In Exp. 1, 12 medium-framed beef heifers (average BW = 389 kg) were used in three simultaneous 4 x 4 Latin square intake trials to evaluate the effects of dietary NDF supply from alfalfa hay, sudan hay, wheat straw, or cottonseed hulls fed in each Latin square at 5, 10, or 15% of dietary DM. Within each roughage concentration, roughage NDF accounted for the majority of variation in NEg intake/kg of BW0.75 among the roughage sources. Averaged across roughage concentrations, NEg intake/kg of BW0.75 tended to be greater (P < 0.10) when heifers were fed cottonseed hulls, sudan hay, or wheat straw than when they were fed alfalfa. In Exp. 2, six medium-framed beef heifers (average BW = 273 kg) were used in a 3 x 3 Latin square design to determine whether diets containing 10% (DM basis) alfalfa, cottonseed hulls, or sudan silage differed in eating rate. Average DM eating rates did not differ (P > 0.10) among roughage sources, which we interpreted to suggest that 90% concentrate diets containing alfalfa, cottonseed hulls, and sudan silage do not differ in the amount of chewing required during eating. In Exp. 3, 105 medium-framed beef heifers (average BW = 275 kg) were used in a 140-d finishing trial to evaluate three methods of dietary roughage exchange. Alfalfa at 12.5% of the dietary DM (ALF12.5) was used as a standard, and cottonseed hulls and sudan silage were each fed at three different levels: exchanged with ALF12.5 on an equal percentage DM basis, an equal NDF basis, or an equal NDF basis, where only NDF from particles larger than 2.36 mm (retained NDF) were considered to contribute to the NDF. No differences (P > 0.10) in ADG, DMI, gain:feed ratio, or NEg intake/kg of BW0.75 were detected between alfalfa and cottonseed hulls exchanged on an equal NDF basis. For sudan silage, exchanging with ALF12.5 on an equal retained NDF basis resulted in no differences (P > 0.10) in ADG, DMI, or NEg intake/kg of BW0.75. These data provide a preliminary indication that depending on the roughage sources evaluated, roughage NDF content and(or) roughage NDF from particles larger than 2.36 mm might provide a useful index of roughage value in high-concentrate finishing diets. PMID- 12078719 TI - Efficacy of intramuscular treatment of beef cows with oxytetracycline to reduce mastitis and to increase calf growth. AB - Spring-calving multiparous Angus x Hereford cows were used to determine the efficacy of intramuscular treatment with oxytetracycline to reduce the incidence of mastitis-causing bacteria, decrease milk somatic cell counts (SCC), and increase calf growth. During 2 yr, milk samples were collected from each quarter from a total of 319 cows at 8 to 14 d after calving and at weaning, to determine the presence of bacteria and SCC. A California mastitis test (CMT) was performed on milk from each quarter of each cow at the initial sample collection. Cows with a CMT score of 1, 2, or 3 in at least one quarter, were randomly assigned to receive either an intramuscular injection of oxytetracycline (n = 63) or the control vehicle (n = 60), and cows with a CMT score of 0 or trace in all four quarters were not treated (n = 196). Calf weights were determined at birth, early lactation, and weaning. The number of somatic cells in milk and the percentage of quarters that were infected increased as CMT score increased (P < 0.01). The presence of mastitis-causing bacteria at calving increased (P < 0.05) the incidence of infection at weaning. The presence of mastitis-causing bacteria at weaning was associated with increased SCC for quarters and average SCC for cows (P < 0.01). Average SCC per cow at weaning increased (P < 0.05) as the number of infected quarters per cow increased. Treatment did not alter (P > 0.10) the percentage of cows or quarters infected with mastitis-causing bacteria or SCC of cows or quarters at weaning. Average SCC per cow was negatively correlated (P < 0.05) with calf weights at early lactation, but not with weaning weights of calves. Treatment did not influence (P > 0.10) calf weights at early lactation or at weaning. Cows with one or more dry quarters after calving had calves that weighed less at early lactation and weaning than cows with four functional quarters (P < 0.01). Intramuscular oxytetracycline treatment of beef cows that had CMT scores of 1 or greater after calving did not reduce intramammary infection rates or increase calf weights at weaning. PMID- 12078720 TI - Effect of creep feed consumption on individual feed intake characteristics and performance of group-housed weanling pigs. AB - To assess the effects of creep feed consumption on individual feed intake characteristics and performance of group-housed weaned pigs, 16 litters (149 piglets) were fed a commercial creep feed (3,040 kcal NE/kg, 15.2 g lysine/kg) supplemented with 1% chromic oxide. Another five litters (48 piglets) were not given access to creep feed (no-feed). Piglets were weaned at 28 d after birth. On d 18, 22, and 27 of age, fecal samples from all the piglets were taken using fecal loops. A green color of the feces indicated that the piglet had eaten creep feed. Piglets that had green-colored feces three times were considered as eaters. Piglets that never showed green-colored feces were considered as non-eaters. At weaning 22 piglets of each type (no-feed, non-eaters, and eaters) were selected based on BW, litter origin, and sex. These 66 pigs were assigned to six pens equipped with computerized feeding stations. Eaters, non-eaters, and no-feed pigs were equally divided over all six pens. After weaning a prestarter (d 0 to 13) and a starter diet (d 14 to 34) were offered for ad libitum consumption. The individual feed intake characteristics of latency time (interval between weaning and first feed intake) and initial feed intake (intake during the first 24 h following first feed intake) and performance traits were determined for all piglets. The pigs that were designated as eaters needed less time between weaning and first feed intake than the pigs that were designated as non-eaters and no feed pigs (P = 0.04 and P = 0.06, respectively). Initial feed intake was not affected (P > 0.1) by feed intake prior to weaning. However, during d 0 to 8 the eaters had more visits per day during which feed was consumed than both the non eaters and no-feed pigs. Averaged over the first 8 d after weaning, the ADFI and ADG of the eaters were higher than that of the non-eaters and no-feed pigs (P < 0.05). Averaged over the total 34-d period the effect of creep feed intake on postweaning ADFI was much less pronounced (P = 0.20), whereas ADG of the eaters was the highest (P < 0.05). Creep feed intake during the sucking period stimulates early postweaning feed intake as well as postweaning performance. PMID- 12078721 TI - Two on-farm data collection methods to determine dynamics of swine compositional growth and estimates of dietary lysine requirements. AB - An experiment was conducted to evaluate the use of two real-time ultrasound data collection methods to develop a dynamic assessment of live weight growth, protein and lipid accretions, and lysine requirement curves on two commercial swine operations. For the first method, pigs (40 barrows and 40 gilts) were weighed (ranging from 18 to 121 kg) and scanned ultrasonically to collect backfat depth and longissimus muscle area measurements every 3 wk in the finishing facility on two farms (serial method). For the second method, pigs (200 gilts and 200 barrows) of similar corresponding ages on the same two farms were weighed and scanned on 1 d (mass scan) at three different times (February, April, and May). Thirty-two pigs/sex were measured at approximately the same ages as with the serial scans. Pigs on farm 1 grew faster and had smaller backfat depths and larger longissimus muscle areas (P < 0.01) than those on farm 2, irrespective of method. These measurements were used to predict empty-body protein and lipid contents using nonlinear functions, which then were converted to accretion rates and lysine requirements at each BW. Protein accretion (g/d) and daily lysine requirements increased and then decreased for each sex on each farm and were higher on farm 1 than on farm 2. Data from the individual mass scans had larger standard errors for modeled live weight growth than data from the serial scans. Combining data from the three mass scans yielded growth curves with standard errors similar to those for the curves from the serial scans. For the protein accretion curves, the standard errors of the combined mass scans were approximately 20% lower than the standard errors of the serial scans. The standard errors for the modeled lysine:calorie ratio requirement from the serial scans were approximately 1% of the requirement at each BW. These results indicated that either the serial or mass scan data-collection method is a practical means of determining on-farm growth and daily protein and lipid accretion rates, which can be used to determine the farm-specific lysine requirements of growing-finishing pigs. PMID- 12078722 TI - Effect of zeranol or melengestrol acetate (MGA) on testicular and antler development and aggression in farmed fallow bucks. AB - Fifteen yearling fallow bucks were randomly assigned by BW to one of three treatment groups: control (C; n = 5), melengestrol acetate (MGA; n = 5), and zeranol (Z; n = 5), to evaluate effects on testicular development, aggressive behavior, antler growth, sexual activity, ADG, and BW. Zeranol-treated bucks received zeranol ear implants (36 mg) at 90-d intervals, and MGA-treated bucks received MGA in the ration (100 microg x animal(-1) x d(-1)). Bucks grazed ryegrass/Coastal bermudagrass pasture and were supplemented with 3:1 corn/soybean meal at 0.45 kg x animal(-1) x d(-1). Body weights, body condition scores (BCS), blood samples, and testis measurements were obtained at d 0 and at 14-d intervals for 229 d. As bucks reached hard antler (7/15 to 8/25), antlers were harvested and weighed, and ejaculates were collected at 14-d intervals. Aggression was evaluated using 10-min video sessions scoring body blows, avoidance, head pushes, and head bunts. Scrotal circumference (SC) and paired testis volume were affected by a day x treatment interaction (P < 0.01); testes of zeranol-treated bucks were smaller than those of control or MGA-treated bucks. First sperm in the ejaculate tended to be delayed (P < 0.10) in zeranol-treated bucks compared with control and MGA-treated bucks. Melengestrol acetate-treated bucks had a maximum sperm concentration in the ejaculate that was three times (P < 0.05) that of control bucks and nine times (P < 0.05) that of zeranol-treated bucks. Antler weight was the least (P < 0.01) for bucks receiving zeranol and greatest (P < 0.10) for MGA treated bucks; intermediate values were recorded for the control bucks. Aggressive behavior was delayed (P < 0.05) for zeranol-treated bucks until treatment effects were overcome. Melengestrol acetate-treated bucks had decreased (P < 0.01) aggressive behavior compared with control bucks. Melengestrol acetate treated bucks had increased (P < 0.05) serum testosterone concentrations compared with control and zeranol-treated bucks. Human chorionic gonadotropin-stimulated peak serum testosterone concentrations for zeranol-treated bucks were delayed (P < 0.01) compared with control and MGA-treated bucks. Although zeranol-treated bucks overcame treatment effects, they were never able to reach testicular measurements or sperm concentrations equal to those of the control or MGA-treated bucks. Zeranol and MGA treatments may have both positive and negative effects that can be utilized when producing slaughter bucks. PMID- 12078723 TI - Effects of double stocking and weighing frequency on pig performance in wean-to finish production systems. AB - Two studies were carried out in different wean-to-finish barns to determine the effects of double stocking on pig growth performance. In Study 1, pigs (n = 1,560) were used in a randomized complete block design with a 2 x 2 factorial arrangement of treatments: initial stocking treatment (Single [52 pigs/pen] vs Double [104 pigs/pen] stocked for 10 wk after weaning) and weighing frequency (High [12 times during the study] vs Low [3 times]) on pig performance from weaning (5.9+/-0.01 kg BW; 17 d of age) to harvest (114+/-0.67 kg BW). Floor and feeder space per pig were 0.650 m2 and 4 cm and 0.325 m2 and 2 cm for the single- and double-stocked treatments, respectively. In Study 2, pigs (n = 1,458) were used in a randomized complete block design to evaluate two initial stocking treatments (Single [27 pigs] vs Double [54 pigs] stocked for 10 wk after weaning) on pig performance from weaning (4.8+/-0.01 kg BW; 15 d of age) to harvest (24 wk after weaning). Floor and feeder space per pig were 0.640 m2 and 3.4 cm and 0.320 m2 and 1.7 cm for single- and double-stocked pens, respectively. In both studies, double-stocked pigs were split at the end of wk 10 into two equal-sized groups of similar mean BW and CV of BW, and one group was moved to a different pen in the same building. In Study 1, performance was not affected (P > 0.10) by frequency of weighing. For the first 10 wk after weaning, the Double compared to the Single treatment had lower ADG (7.7 and 7.9%, for Studies 1 and 2, respectively; P < 0.001) and lighter pigs at wk 10 (6.8 and 7.3%, respectively; P < 0.001). During the first 10 wk in Study 1, Double compared to the Single pigs had lower ADFI (7%; P < 0.001) but similar gain:feed (P > 0.10). From wk 11 to harvest, pigs on Double and Single treatments had similar (P > 0.10) ADG in both studies and, in Study 1, ADFI was unaffected by initial stocking treatment, but double-stocked pigs had greater gain:feed (4%, P < 0.01). Double-stocked pigs required an additional 2 d to reach a fixed harvest BW (P < 0.05) in Study 1 and were lighter (4%; P < 0.05) at 24 wk after weaning in Study 2. Carcass measures were similar (P > 0.10) for double- and single-stocked pigs. Double-stocked pigs that were moved at the end of 10 wk had growth performance similar (P > 0.10) to those that remained in the original pen. In summary, double stocking reduced growth rate to 10 wk after weaning but subsequently had no effect on growth rate and improved feed efficiency. PMID- 12078724 TI - Effects of stocking rate and crude protein intake during gestation on ground cover, soil-nitrate concentration, and sow and litter performance in an outdoor swine production system. AB - Pregnant gilts (n = 126) were assigned randomly to 12 0.4-ha old world-spar bluestem (Bothriochloa ischaemum) pastures in an outdoor swine (Sus scrofa) production system to examine effects of stocking rates (17.5 or 35 gilts/ha; 7 or 14 gilts per pasture) and dietary N on percentage of ground cover, soil nitrate (NO3-) concentration, and reproductive performance. Treatments were arranged factorially with two stocking rates and two diets equivalent in dietary lysine but different in CP (control = 14.7% CP vs experimental = 12.6% CP) with three pastures per treatment. The experiment was repeated during a second parity with the same animals on the same treatments. Each triangular gestation pasture was subdivided into three regions: 1) near the point or radial center; 2) the middle region that contained a hut and a wallow area; and 3) the outer section where gilts were fed each day. Soil samples (15 cm deep) were taken at the beginning and end of the 306-d study, and soil nitrate-N concentrations were determined. Percentage of ground cover was visually estimated initially and every 30 d thereafter through d 306. Before farrowing, gilts were moved to identical pastures for farrowing and were fed a common 16% CP sorghum (Sorghum bicolor) based lactation diet beginning at the time of movement to the farrowing pasture. Pregnant gilts were weighed at the time of assignment to treatments in the gestation pastures, when they were moved to farrowing pastures, and at weaning. Production data included total number of pigs born per sow, number of pigs born alive or dead, average birth weight, number of pigs weaned, average weaning weight, and mortality. No differences (P > 0.05) were observed between treatments in soil NO3- concentrations. Percentage of ground cover was decreased (P < 0.01) by the higher stocking rate when grazing was initiated in March/April but recovered rapidly after removal of pigs. More (P < 0.01) pigs were weaned per sow (8.4 vs 7.1+/-0.34) from higher gestation-stocking rate groups. Pig mortality in farrowing was greater (P < 0.05) for lower gestation-stocking rates (25.7% vs. 18.1+/-1.9%). A stocking rate of 35 sows/ha might have increased production potential but was associated with a rapid loss of ground cover during spring. PMID- 12078725 TI - Effects of breeding date, weaning date, and grazing season length on profitability of cow-calf production systems in southeastern Montana. AB - Production data from 11 southeastern Montana ranches were used to parameterize a bio-economic computer model of cow/calf range production. Effects of changes in breeding date, weaning date, and range removal date on system performance for a ranch with a fixed forage resource base (3,060 animal unit months of range forage and 744 t of hay) were simulated. Input costs were locally established in 1994. Cattle prices were determined by week from 13-yr averages. For the base scenario, breeding season was 66 d with breeding starting on June 9. Weaning, range removal, and calf sales occurred on November 3. Cows were fed stored forages from November 3 until turnout to grazing (May 1). Five replications were simulated for combinations of breeding, weaning, and range removal dates in a factorial design. Each factor was deviated from the base scenario by +/- 14 and 28 d. Production efficiency was measured by break-even steer price. Gross margin (gross revenue - variable costs) was used as a measure of profitability. Increasing calf age (and weight) at sale time, by decreasing breeding date and(or) increasing weaning date, improved ranch efficiency and profitability. Increasing range removal date improved system performance even though extending the grazing season led to decreased herd size. Compared to the base system, the best system increased gross margin by approximately 17%. Responses for gross margin reflect the dynamics of herd size, purchased feed expense, and production efficiency. Results suggest that for range-based cow-calf enterprises in the northern Great Plains, production efficiency and profitability may be improved by increasing calf weaning age and extending the grazing season, even if herd size must be reduced. PMID- 12078727 TI - Heterozygosity for genes influencing a quantitative trait. AB - Choosing families to sample for a quantitative trait locus mapping experiment is a critical component of experimental design because only heterozygous families contribute information to the analysis. Additive genetic variance of a paternal half-sib family can be partitioned into two parts: a variance component of maternal source that is constant across different families and a variance component of paternal source that is defined as an index of heterozygosity of a sire. This index is shown to be an upper limit of variance among marker genotypes of a half-sib family and can be used to identify highly heterozygous sires, thus improving the power of detecting QTL in detection studies. Simulated progeny phenotypic data were used to estimate sire's heterozygosity index via an ANOVA method, and accuracy of the estimation was evaluated with the correlation coefficient between the true and estimated index summarized both as the correlation and by the correct ranking of results as measured by the ratio of the true average heterozygosity index of experimentally selected parents to average heterozygosity of all sires. Positive but small correlation can be achieved in the estimation of a sire's heterozygosity when based on the daughters' phenotypic data, and accuracy was improved when progeny-tested sons were used to estimate their grandsire's heterozygosity index, depending on the genetic model of a trait and the size and structure of families. PMID- 12078726 TI - Maternal effects on traits measured during postweaning performance test of swine from four breeds. AB - The objective of this study was to investigate the importance of maternal genetic effects on postweaning performance traits of Yorkshire, Landrace, Duroc, and Hampshire breeds of swine. Data consisted of performance test records collected in a commercial swine operation from 1992 to 1999. Boars from 60% of the litters were culled at weaning based on a combination of maternal and performance indexes that differed by breed. Remaining boars and all females were grown to 100 d of age. At this time all pigs were weighed (WT100) and selected for testing using recalculated breed-specific indexes (n = 15,594, 55,497, 12,267, and 9,782 for Landrace, Yorkshire, Duroc, and Hampshire, respectively). All pigs were weighed at the end of the 77-d test, and backfat (BF) and loin eye area (LEA) were measured over the 12th rib by ultrasound. Average daily feed intake was calculated for boars, and ADG was calculated for all animals. Genetic parameters were estimated for each breed and trait using multiple-trait DFREML procedures. Fixed effects were contemporary groups and either initial or final test age as a covariate. Four models were examined. Model 1 included only the additive genetic effect of the animal. Model 2 added the common litter environmental effect; Model 3 added the maternal genetic value assumed to be uncorrelated with additive genetic effects. Model 4 was the same as Model 3 with additive and maternal genetic effects assumed to be correlated. All models were two-trait models with WT100 as the second trait. Ratios of likelihoods were used to compare models. Maternal effects were important (P < 0.05) for WT100, ADG, ADFI, LEA, and BF in Landrace; for WT100, ADG, LEA, and BF in Yorkshire; for WT100 and ADG in Duroc, and for WT100 in Hampshire. Estimates of heritabilities for direct additive effects using the appropriate model for ADG, ADFI, LEA, and BF were 0.28, 0.34, 0.48, and 0.63 for Landrace; 0.26, 0.31, 0.39, and 0.65 for Yorkshire; 0.14, 0.20, 0.26, and 0.35 for Duroc; and 0.17, 0.23, 0.25, and 0.31 for Hampshire, respectively. Heritability estimates for maternal genetic effects for ADG, ADFI, LEA, and BF were 0.02, 0.05, 0.06, and 0.07 for Landrace and 0.02, 0, 0.04, and 0.06 for Yorkshire, respectively. They were zero for all traits except ADG (0.03) in Duroc and for all traits in Hampshire. Maternal effects may need to be considered in genetic evaluation of performance traits in some breeds of swine. PMID- 12078728 TI - Genetic and environmental parameters for steer ultrasound and carcass traits. AB - Carcass measurements for weight, longissimus muscle area, 12-13th-rib fat thickness, and marbling score, as well as for live animal measurements of weight at the time of ultrasound, ultrasound longissimus muscle area, ultrasound 12-13th rib fat thickness, and ultrasound-predicted percentage ether extract were taken on 2,855 Angus steers. The average ages for steers at the time of ultrasound and at slaughter were 391 and 443 d, respectively. Genetic and environmental parameters were estimated for all eight traits in a multivariate animal model. In addition to a random animal effect, the model included a fixed effect for contemporary group and a covariate for measurement age. Heritabilities for carcass weight, carcass longissimus muscle area, carcass fat thickness, carcass marbling score, ultrasound weight, ultrasound longissimus muscle area, ultrasound fat thickness, and ultrasound-predicted percentage ether extract were 0.48, 0.45, 0.35, 0.42, 0.55, 0.29, 0.39, and 0.51, respectively. Genetic correlations between carcass and ultrasound longissimus muscle area, carcass and ultrasound fat thickness, carcass marbling score and ultrasound-predicted percentage ether extract, and carcass and ultrasound weight were 0.69, 0.82, 0.90, and 0.96, respectively. Additional estimates were derived from a six-trait multivariate animal model, which included all traits except those pertaining to weight. This model included a random animal effect, a fixed effect for contemporary group, as well as covariates for both measurement age and weight. Heritabilities for carcass longissimus muscle area, carcass fat thickness, carcass marbling score, ultrasound longissimus muscle area, ultrasound fat thickness, and ultrasound predicted percentage ether extract were 0.36, 0.39, 0.40, 0.17, 0.38, and 0.49, respectively. Genetic correlations between carcass and ultrasound longissimus muscle area, carcass and ultrasound fat thickness, and carcass marbling and ultrasound-predicted percentage ether extract were 0.58, 0.86, and 0.94, respectively. The high, positive genetic correlations between carcass and the corresponding real-time ultrasound traits indicate that real-time ultrasound imaging is an alternative to carcass data collection in carcass progeny testing programs. PMID- 12078729 TI - Two-step and random regression analyses of weight gain of station-tested beef bulls. AB - Our objectives were to compare a two-step model and a joint procedure via random regression model for evaluating weight gain of beef bulls, weighed every 28 d on 140-d test, and to estimate genetic, environmental, and phenotypic parameters. Two-step analysis consisted of fitting fixed linear regressions to weights of each bull to determine weight gain on test. In the second step, gain on test was analyzed by a mixed model that included fixed effects of breed, test group, and starting age and random effects of weaning herd-year group and animal (additive genetic). The random regression model included the same effects as the two-step mixed-model analysis with an additional random animal permanent environment effect. Fourth-order Legendre polynomials of days on test were fitted for all fixed and random effects in the random regression model, except for breed. Breed effects and residual variances varied for each measurement period. Variance components and EBV for gain were obtained from the covariance function and estimates of random regression coefficients for weight, respectively. Random regression heritability estimates for gain on test increased over time, being maximum at end of test (0.38) and equal to two-step estimate. Permanent environment variance ratio estimates also increased over time and were greater than heritability estimates. Estimate of weaning herd-year variance ratio was approximately constant over time, being equal to 0.07 at end of test and similar to two-step estimate. Genetic correlations between gain through different periods on test given by random regression model were high (from 0.81, between 28 and 140 d gain on test, to 0.99, between 112 and 140-d gain on test). Genetic correlations between gain on discrete 28-d intervals were moderate to high (e.g., 0.49 and 0.99 between the last 28 d on test and the first and fourth 28 d, respectively). Rank correlations between EBV for 140-d gain by the two procedures were 0.98, 0.84, and 0.73 for all bulls and the 5% and 1% of bulls with highest random regression EBV, respectively. Results indicated that the two procedures rank top bulls quite differently for 140-d gain on test. Random regression model accounted for changes over time of genetic and environmental effects on the test weight gain curve of the bulls. Use of 112-d instead of a 140-d test provided similar ranking of bulls on the basis of EBV for gain on test. PMID- 12078730 TI - Heritability and repeatability of sexual performance scores of rams. AB - Sexual performance has been subjectively measured with a libido test during screening of rams before public sale and breeding at the U.S. Sheep Experiment Station from 1990 to 2000. The objective of this study was to determine whether sexual performance was genetically influenced. Sexual performance scores ranged from 1 to 6 with scores increasing from sexually inactive to highly sexually active in the presence of estrous ewes. The overall average score was 3.5+/-0.02. Records from four breeds (Columbia, n = 807; Polypay, n = 1,668; Rambouillet, n = 1,208; and Targhee, n = 1,002) were combined into one analysis because breeds had similar phenotypic variances. Total number of records was 4,685, which included a second sexual performance test on 1,212 rams in the following year. Variance components were estimated using a REML algorithm. Fixed effects were breed of ram, selection line within breed, and year by breed. A permanent environmental effect for ram was included to account for repeated observations on individual animals. Age and weight of the rams at time of the libido test were linear covariates and were breed-specific. Adjusted means for sexual performance scores did not differ among breeds (P > 0.05). Age was a significant effect (P < 0.01), with sexual performance score increasing 0.05 units for each month of age. The additive genetic variance was estimated as 0.54. The estimate of variance due to ram permanent environmental effects was 1.19. The residual variance was estimated to be 0.67. The heritability estimate was moderate (0.22+/-0.04) and repeatability was high (0.72). These results imply that one screening for sexual performance provides a reliable measure of sexual performance and that favorable response to selection for ram serving capacity may be expected. PMID- 12078731 TI - Effects of dietary ingredients on manure characteristics and odorous emissions from swine. AB - Two feeding studies were conducted to examine the impact of dietary inclusion of specific feed ingredients on manure characteristics and manure odor. In one study, 72 finishing pigs were used to evaluate the effects of distillers dried grains with solubles (DDGS) on pig performance, manure characteristics, and odorous emissions. Three diets containing 0, 5, and 10% DDGS were fed during six 4-wk feeding periods. Week 1 served as a dietary adjustment period. Animals were housed in two feeding rooms (six pigs/room) with one treatment/room. A new group of animals (average initial BW = 85.8 kg) was used for each feeding period. Diets were replicated four times. Rooms were equipped with individual shallow manure storage pits that were cleaned once weekly (d 7). On d 4 and 7 of each week, manure pit samples, for chemical analyses, and air samples, for olfactometry analysis, were collected from each room. Odor dilution threshold was greater on d 7 than on d 4 of manure storage across all treatments (P < 0.01). No treatment differences in manure composition were noted. In the second study, weaned pigs (approximately 5 wk old) were fed isonitrogenous diets containing 0, 1.5, or 3% bloodmeal. Pigs were housed by diet (three pigs/diet) in one of four individual feeding rooms. A new group of pigs was used for each of the two, 4-wk feeding periods. During period 1, the 3% bloodmeal diet was fed in two of the four rooms; the 0% bloodmeal diet was fed in two rooms during period 2. Manure samples, for chemical analyses, and air samples, for olfactometry analysis, were collected 2 d per week (d 4 and d 6) from each room during wk 2 through 4. No significant treatment differences were observed for odor dilution threshold (P = 0.30). Longer manure storage time, 6 d vs 4 d, resulted in a larger odor dilution ratio (P < 0.01). Manure composition was unaltered by storage time. Results suggest that odor intensifies during storage. PMID- 12078732 TI - Management of the female stimulus during semen collection and its association with libido re-establishment and semen characteristics of goats. AB - The effect of management of the female stimulus during semen collection and its association with libido re-establishment and the semen characteristics of sexually mature goats was evaluated. Eight 2-yr-old bucks were individually subjected to 12 weekly trials in which one of the following three treatments was applied alternately. In treatment one, each male performed two ejaculations on an estrus-induced, restrained doe and semen was collected with an artificial vagina (AV). In treatment two, males were collected as in the former treatment, but a different doe served as the stimulus female for the second ejaculation. In treatment three, two restrained does were present while collecting each buck, allowing males to choose which female to mount and serve in each of the two collections. Number of mounts before first ejaculation and reaction times (period from introduction of the ram to the test arena to his first ejaculation) were similar among treatments (P > 0.05). A significant (P < 0.05) decrease was found for number of mounts performed before achieving the second ejaculation and for latency between ejaculations in the third treatment. No differences (P > 0.05) were found for semen volume (0.8+/-0.03, 1.0+/-0.03, and 0.9+/-0.02 mL) or sperm concentration (4.4+/-0.15, 4.6+/-0.13, and 4.4+/-0.86 spermatozoa/mL) among the three treatments or between the first and second collection. However, a trend to decrease volume and concentration was observed. It was concluded that the simultaneous presence of two females while bucks are collected reduced the number of mounts performed before the second ejaculation and the latency between ejaculations. PMID- 12078733 TI - Technical note: fourier transform infrared (FTIR) spectroscopy as an optical nose for predicting odor sensation. AB - Quantifying odor is important for objectively assessing the impact of animal production systems on surrounding areas. A possible method that has received little attention is Fourier transform (mid) infrared spectroscopy (FTIR). Gases that contribute to odor have unique infrared spectra, and the advantage of FTIR over electronic nose technology or gas chromatography is that theoretically all these gases can be analyzed instantaneously. To determine the feasibility of FTIR for predicting odor, 71 air samples analyzed by olfactometry were scanned in a spectrometer using an 84-m path-length gas cell. Scans were obtained over a period of about 1 min and from 4,000 to 740 cm(-1) with a resolution of 0.5 cm( 1). Calibrations for predicting odor were developed using partial least squares regression with full cross-validation. Air samples were obtained from experiments with pigs fed diets formulated to alter odor emission or from stored manure. Odor threshold dilution ratios averaged 676+/-491 units, with a range from 120 to 2,161. Using these samples, a prediction error for odor sensation of 344 units (R2 = 0.51) was obtained. Log transformation of the odor data improved the R2 to 0.61. Based on the olfactometry data, it is estimated that the measurement error of olfactometry is 250 units, which limits the R2 of any method to approximately 0.74. Thus, this calibration is very encouraging. In conclusion, FTIR shows promise as a practical means for objectively assessing swine odor. PMID- 12078734 TI - Effect of nutrient intake on the development of the somatotropic axis and its responsiveness to GH in Holstein bull calves. AB - We investigated the effect of increasing nutrient intake on the responsiveness of the GH/IGF-I system in calves fed a high-protein milk replacer. Fifty-four Holstein bull calves were fed one of three levels (low, medium, and high; n = 18 per treatment) of a 30% crude protein, 20% fat milk replacer to achieve target rates of gain of 0.50, 0.95, or 1.40 kg/d, respectively, for low, medium, and high. Six calves per treatment were slaughtered at approximately 65, 85, and 105 kg BW. Additionally, six calves were slaughtered at 1 d of age to provide baseline data. Plasma aliquots from blood samples collected weekly were analyzed for IGF-I, insulin, glucose, NEFA, and plasma urea nitrogen (PUN). Plasma IGF-I and insulin, measured weekly, increased (P < 0.001) with greater nutrient intake from wk 2 of life to slaughter. Plasma glucose and NEFA also increased (P < 0.05) with nutrient intake. In addition, each calf underwent a GH challenge beginning 4 d before the scheduled slaughter. Plasma from blood collected before the first GH injection and 14 and 24 h after the third injection was analyzed for IGF-I and PUN. Response to challenge, calculated as the absolute difference between the prechallenge and 14-h postchallenge plasma IGF-I concentrations, was significant in calves on all three treatments. Plasma urea nitrogen was not different among treatments as measured weekly but decreased (P < 0.001) following GH challenge in all calves. Results of ribonuclease protection assays showed increased expression of hepatic mRNA for GH receptor 1A and IGF-I with increased intake. The amounts of GH receptor and IGF-I mRNA in muscle and adipose, however, were not affected by intake. In summary, plasma IGF-I was elevated in calves with increased nutrient intake, and the elevations in plasma IGF-I following short-term administration of GH were significant in all calves by 65 kg BW. Data demonstrate that in well-managed milk-fed calves the somatotropic (GH/IGF-I) axis is functionally coordinated and sensitive to nutrient intake and GH. PMID- 12078736 TI - Growth performance and metabolic and endocrine traits in calves pair-fed by bucket or by automate starting in the neonatal period. AB - Effects of different feeding frequencies on growth performance, preprandial metabolic parameters, and endocrine traits were studied in calves up to 28 d after birth and respective postprandial changes were investigated on d 3 and 7. Calves were fed with a computer-programmed automate that allowed frequent daily intakes (GrA; n = 7) or were fed twice daily by bucket (GrB; n = 7) the same daily amounts (pair-feeding) of colostrum and milk. Weight gains did not differ significantly between groups. Plasma protein was higher (P < 0.01) from d 14 to 28 in GrB than in GrA. Plasma glucose increased postprandially in GrB on d 3 and 7 but in GrA only on d 3, and there was a significant treatment x time interaction on d 3 and 7 after feed intake. For plasma triglycerides there was a significant postprandial treatment x time interaction on d 3, and triglycerides were higher (P < 0.05) in GrA than in GrB before feed intake on d 7. For insulin concentrations on d 7 the treatment x time interaction was significant and concentrations 8 h after feed intake were higher (P < 0.05) in GrA than GrB. For concentrations of growth hormone on d 7 during an 8-h period the treatment x time interaction was significant and concentrations from 320 to 340 min after feed intake were higher (P < 0.05) in GrB than in GrA. Plasma IGF-I concentrations were higher (P < 0.05) in GrA than in GrB on d 7. The hematocrit and concentrations of immunoglobulin G, albumin, urea, creatinine, L-lactate, nonesterified fatty acids, cholesterol, glucagon, and thyroxine did not differ between groups. In conclusion, feed intake at high frequency by an automate transiently changed some metabolic and endocrine traits (glucose, triglyceride, insulin, and IGF-I) but had no significant effects on growth performance during the first 4 wk of life. PMID- 12078735 TI - Developmental changes in ketogenic enzyme gene expression during sheep rumen development. AB - Ketogenesis is the conversion of acetyl-CoA to the ketone bodies acetoacetate and beta-hydroxybutyrate (BHBA). In hepatic ketogenesis, which occurs during fasting in both nonruminant and ruminant animals, the source of acetyl-CoA is the mitochondrial oxidation of predominantly long-chain fatty acids. In the mature, fed ruminant animal, the ruminal epithelium is also capable of producing ketone bodies. In this case, the source of acetyl-CoA is the mitochondrial oxidation of butyrate produced by the microbial fermentation of feed. The purposes of this study were to determine ontogenic and dietary effects on ketogenic enzyme gene expression in developing lamb ruminal epithelium. Twenty-seven conventionally reared lambs and twenty-seven milk-fed lambs were slaughtered between 1 and 84 d of age. Six additional milk-fed lambs were weaned (the fed group) or maintained on milk replacer with a volatile fatty acid gavage (the VFA group) until 84 d of age. At slaughter, total RNA was extracted from samples of ruminal epithelium. The expression of the genes encoding acetoacetyl-CoA thiolase, the first enzyme in the ketogenic pathway, and 3-hydroxy-3-methylglutaryl-CoA synthase, the rate limiting enzyme in the ketogenic pathway in nonruminant liver, were examined. Acetoacetyl-CoA thiolase and 3-hydroxy-3-methylglutaryl-CoA synthase mRNA concentrations increased with age independent of diet. 3-Hydroxy-3-methylglutaryl CoA synthase mRNA levels in ruminal epithelium obtained from milk-fed lambs were low before 42 d of age, but a marked increase occurred by 42 d of age. At 84 d of age, there were no differences in acetoacetyl-CoA thiolase and 3-hydroxy-3 methylglutaryl-CoA synthase expression due to diet. The pattern of the expression of these genes, in particular, 3-hydroxy-3-methylglutaryl-CoA synthase, parallels the rate of production of BHBA by rumen epithelial cells isolated from the same lambs, which increased to conventionally reared adult levels at 42 d of age and did not differ with diet. In conclusion, development of the ketogenic capacity of the ruminal epithelium occurs as the animal ages, regardless of dietary treatment. Thus, the expression of the genes encoding the ketogenic enzymes are not affected by the presence of VFA in the ruminal lumen. PMID- 12078737 TI - Grass silage versus maize silage effects on retail packaged beef quality. AB - The effects of three preslaughter diets on heifer beef quality were investigated. Heifers (n = 45) were divided into three groups and fed for ad libitum consumption either maize silage, grass silage, or a 50:50 mixture of maize silage and grass silage. Meat quality was determined by measuring color, lipid oxidation, alpha-tocopherol levels, and fatty acid composition. Beef from the maize silage group had poorest color stability (P < 0.05), whereas beef from the grass silage diet had best (P < 0.05) color stability. The visual panel least preferred the maize silage group after 2 or more days of display, and lipid oxidation was significantly (P < 0.001) higher in this group compared to the 50:50 maize:grass silage and grass silage groups. There was a significant (P < 0.001) difference in the alpha-tocopherol levels detected in the meat from the three dietary groups. Alpha-tocopherol levels increased in the order: maize silage < 50:50 maize:grass silage < grass silage, at levels of 2.08, 2.95, and 3.84 microg/g meat, respectively. Fatty acid analysis indicated 18:3 was significantly (P < 0.001) lower in the maize silage-fed group than in the maize:grass silage and grass silage groups. However, 18:3 was significantly (P < 0.001) higher in the grass silage group than in the other two groups. There were no significant differences in all other fatty acids among the three dietary groups. It was concluded that beef from grass silage-fed animals had better overall quality in terms of color, lipid oxidation, and alpha-tocopherol levels than beef from maize silage fed animals. PMID- 12078738 TI - Effect of dietary energy supply and fat source on the fatty acid pattern of adipose and lean tissues and lipogenesis in the pig. AB - Forty Large White barrows were used to determine whether the effects of dietary fat source (tallow or soy oil at 5% of the diet) on lipogenesis and fatty acid profile of porcine adipose and lean tissue were dependent on dietary digestible energy density (8.8 vs 14.0 MJ DE/kg). Barrows were allocated to one of four groups and offered a fixed amount of feed (170 g x BW0.569/d) from 27 to 105 kg BW. The fatty acid composition of the backfat layers (BF), omental fat (OF), and i.m. adipose tissue of longissimus muscle as well as the activity of lipogenic enzymes of the adipose tissues were determined. Growth performance and carcass characteristics were affected by the dietary energy level (P < 0.01) but not by fat source. In accordance with the lower carcass fat deposition, the activity of lipogenic enzymes were decreased in the low-energy groups (P < 0.01). Within dietary energy level, inclusion of soy oil resulted in increased proportion of PUFA that was compensated by decreased saturated (SFA) and monounsaturated fatty acid (MUFA) proportions (P < 0.01). The SFA changes accounted for 23 (BF) and 24% (OF) of the PUFA changes in the high-energy and 31 (BF) and 39% (OF) in the low energy diets. The differences in the fatty acid proportions between the soy oil and tallow group were more pronounced in the low-energy groups (fat source x energy density interactions: P < 0.01). Pigs fed the soy oil, low-energy diet had decreased SFA (BF: 28%; OF: 30%) and MUFA (BF: 13%; OF: 19%) concentration, whereas PUFA concentration was increased (BF: 59%; OF: 88%) compared with pigs fed the soy oil, high-energy diet. However, in the tallow groups, pigs fed the low-energy diets had slightly decreased SFA (BF: 14%; OF: 12%) and relatively constant MUFA (BF: 3%; OF: 1%), whereas PUFA concentration increased (BF: 39%; OF: 62%) relative to pigs fed the tallow high-energy diet. Lipid content of the i.m. adipose tissue was decreased in the low-energy groups (P < 0.05). Contrary to what was observed in the adipose tissues, increased PUFA concentration in the neutral and polar lipid fractions of the longissimus muscle was predominantly compensated by reduced MUFA deposition. In the polar lipid fraction, the proportions of both SFA and MUFA were decreased by the low-energy diet. Thus, the extent to which tissue concentration of fatty acids are altered from dietary fats differing in the degree of unsaturation depends on the dietary energy level. PMID- 12078739 TI - Effects of modified tall oil and vitamin E on growth performance, carcass characteristics, and meat quality of growing-finishing pigs. AB - Crossbred barrows (n = 72) were used to evaluate effects of diet supplementation with modified tall oil (MTO; 0.0 or 0.50%) and vitamin E (0, 22, or 110 IU/kg) on growth performance, carcass traits, and longissimus muscle (LM) quality traits of finishing pigs. Pigs were blocked by ancestry and initial BW and allotted randomly to treatments in a 2 x 3 factorial. Corn-soybean meal-based diets were fed in two phases: 45.5 to 81.6 (1.00% lysine) and 81.6 to 114.6 (0.75% lysine) kg BW with no added fat. From 45.5 to 81.6 kg, pigs fed MTO had greater ADG (P = 0.03) regardless of added vitamin E; otherwise, treatment did not affect growth performance. Carcasses from pigs fed MTO had reduced (P < 0.05) average backfat (2.76 vs 2.92 cm) and firmer bellies compared to those fed no MTO. Boneless loins were cut into 2.54-cm chops at 7 d postmortem and evaluated for display color, thiobarbituric acid-reactive substance (TBARS), Warner-Bratzler shear force (WBSF), and sensory panel ratings. Visual color was similar (P > 0.05) among treatments at 0 and 1 d of display. At 4 and 6 d of display chops from pigs fed MTO with 110 IU vitamin E/kg had less deterioration (P < 0.05) than chops from pigs fed MTO with 0 IU vitamin E/kg and 0.0% MTO with 22 or 110 IU vitamin E/kg. The CIE L*, a*, b* and spectral values also suggested a delay in color deterioration for chops from pigs fed MTO with 110 IU vitamin E/kg. At 6 and 8 d of display, chops from pigs fed 110 IU vitamin E/kg had lower (P < 0.05) L* values than those from pigs fed 0 or 22 IU vitamin E/kg, and higher (P < 0.05) a* values than those from pigs fed 0 IU vitamin E/kg feed. A higher (P < 0.05) %R630/%R580 (indicator of more oxymyoglobin) was observed for chops from pigs fed MTO with 110 IU vitamin E/kg than those from pigs fed 0.0% MTO with 22 or 110 IU vitamin E/kg and MTO with 0 IU vitamin E/kg. Chops from pigs fed MTO with 110 IU vitamin E/kg had lower (P < 0.05) TBARS values than those from pigs fed MTO with 0 IU vitamin E/kg. No differences (P > 0.05) were detected among treatments for WBSF or sensory evaluations. The addition of MTO in swine diets improved belly firmness and reduced backfat, and feeding MTO with high levels of vitamin E extended display life without affecting palatability of LM chops. PMID- 12078740 TI - The impact of longissimus glycolytic potential and short-term feeding of magnesium sulfate heptahydrate prior to slaughter on carcass characteristics and pork quality. AB - The objective of this study was to compare the effects of longissimus glycolytic potential (GP) and of time of feeding of supplemental magnesium sulfate heptahydrate on carcass and pork quality traits. The study was carried out in a 2 x 2 x 4 factorial arrangement; the treatments were sex (castrate vs gilt), GP (Low [normal] vs High), and time of feeding of magnesium sulfate-fortified diets (0 [control] vs 2 vs 3 vs 5 d prior to slaughter). Glycolytic potential was determined on a biopsy sample of longissimus from the live animal prior to the start of the study. A total of 144 pigs were allotted to the feeding-time treatments on the basis of sex (castrate and gilt), weight, and GP. Pigs were placed in individual pens and had free access to water. Prior to the start of the study, pigs were given ad libitum access to a standard finisher diet. During the study, animals were fed at a fixed level of 2.75 kg of a standard finisher diet/day; the fortified diet contained 3.2 g/d of additional magnesium. At the end of the feeding period, animals were transported to a commercial packing facility and slaughtered within 15 min of arrival. Fresh meat quality was measured on the longissimus. There were no treatment interactions. Carcass traits were similar across time of feeding treatments. Backfat thickness at the last lumbar vertebra and 10th rib were lower (P < 0 .05) for High than for Low GP pigs. High GP pigs had lower ultimate pH (P < 0.001) and higher drip (P < 0.05) and purge loss (P < 0.01) than Low GP pigs. Drip loss was reduced (P < 0.05) for pigs fed the magnesium-fortified diet for 5 and 2 but not for 3 d compared to controls (8.98, 7.29, 7.89, and 7.41 for the 0-, 2-, 3-, and 5-d treatments, respectively, SEM 0.447). Purge loss was similar for all of the time of feeding treatments. Longissimus L* values were lower (P < 0.05) for the 2-d treatment than for the controls. Results from this study suggest an inconsistent effect of short-term feeding of magnesium sulfate on muscle color and drip loss in pigs with both Low (normal) and High GP. PMID- 12078741 TI - Determination of true ileal amino acid digestibility and the endogenous amino acid outputs associated with barley samples for growing-finishing pigs by the regression analysis technique. AB - This study was conducted to determine true ileal AA digestibility coefficients and the endogenous AA outputs associated with barley samples for growing finishing pigs using the regression analysis technique with dual digestibility markers. Six barrows, with 30.5 and 58.6 kg average initial and final BW, were fitted with a simple T-cannula at the distal ileum and fed six barley-based diets at close to ad libitum feed intake according to a 6 x 6 Latin square design. The six diets contained 97% of six barley samples varying from low to high in CP and AA contents (8.5, 9.2, 9.8, 11.5, 12.6, and 15.6% CP, respectively, on DM basis). The dietary NDF content ranged from 16.8 to 23.8% on DM basis. Chromic oxide (Cr2O3) and acid-insoluble ash (AIA) were used as digestibility markers. Each experimental period lasted 7 d. Ileal digesta were collected, at 2-h intervals, for a total of 24 h during d 6 and 7. There were linear relationships (P < 0.01) between dietary contents of apparent ileally digestible and total CP and AA as determined by using either Cr2O3 or AIA as a digestibility marker. The use of Cr2O3 vs AIA affected (P < 0.01) the determination of true ileal AA digestibility coefficients and the endogenous CP and AA outputs. However, there were no differences (P > 0.01) in the true ileal AA digestibility coefficients in barley samples between this study and the average values reported in the literature. The endogenous CP and AA outputs determined in this study were higher (P < 0.01) than reported values (35.1+/-3.0 vs 14.7+/-1.1 g CP/kg DMI). It is concluded that dual digestibility markers should be used to measure true ileal AA digestibility coefficients and endogenous AA outputs when dietary fiber content is high and the ileal digesta is collected through a simple T-cannula in the pig. True rather than apparent ileal AA digestibility coefficients determined in barley samples should be used in diet formulation for swine. The gastrointestinal endogenous AA secretion, recycling, and output losses are important in whole-body AA utilization and homeostasis, especially when fiber-enriched diets are fed to growing-finishing pigs. PMID- 12078742 TI - The effects of dietary fat sources, levels, and feeding intervals on pork fatty acid composition. AB - Two experiments investigated the quantitative relationship between dietary fat and fatty acid composition of pork. Experiment 1 was designed to establish the rate of decline for linoleic acid and iodine value of pork fat during the late fattening phase following a dietary reduction. Gilts (n = 288) were fed diets varying in linoleic acid content from 4.11 to 1.56% for 4, 6, or 8 wk prior to slaughter. The maximum rate of decline was 2% 18:2 per week and 2.5 iodine value units per week. Experiment 2 evaluated the effects of dietary fat source and level on carcass fatty acid composition and on pork quality characteristics. Barrows (n = 147) and gilts (n = 147) were allocated to seven dietary treatments for the last 6 wk of the finishing phase. Diets contained 0, 2.5, or 5% dietary fat comprised of 100, 50, or 0% beef tallow. The balance was provided by animal vegetable blended fat. As the level of tallow increased there was a linear decrease (P < 0.05) in 18:2 content and iodine value of carcass fat. Conversely, 16:1 and 18:1 increased linearly (P < 0.05) as tallow increased. However, 16:1 decreased linearly (P < 0.05) as level of fat increased. As the level of tallow was increased a greater reduction in 18:2 and iodine value was observed in diets with 5% dietary fat compared to diets with 2.5% fat (P < 0.05). These results indicate that reduction of dietary PUFA content had the desired effect of lowering 18:2 content and iodine value of pork fat and that significant alterations could be elicited in as little as 6 to 8 wk of feeding. PMID- 12078743 TI - Technical note: epimerization of ergopeptine alkaloids in organic and aqueous solvents. AB - Purified ergopeptine alkaloids are often used in studies related to tall fescue toxicosis without regard to epimerization that occurs when ergopeptines are solvated. The objectives of this study were to measure the rates of alpha ergocryptine epimerization to alpha-ergocryptinine at room temperature and at -40 degrees C, and to measure the rate of ergovaline epimerization to ergovalinine at 37 degrees C. Alpha-ergocryptine tartrate was stable (< 0.5% epimerization) in protic or aprotic solvents when stored at -40 degrees C for 20 to 52 d. At room temperature, alpha-ergocryptine epimerization in chloroform did not occur; epimerization was modest in acetone and acetonitrile (< 5%) but was substantial in methanol (78% by 38 d) and in a 70:30 water methanol mix (47% by 42 d). Ergovaline epimerization to ergovalinine occurred at 37 degrees C in 0.1 M phosphate buffers (pH 3, 7.5, and 9) in 9% aqueous solutions of fetal bovine serum (FBS), and in water, methanol, and acetonitrile. The degree of epimerization at 37 degrees C was solvent-dependent. Epimerization rates with respect to time were roughly linear in phosphate buffer (pH 3 only), water, methanol, and acetonitrile; epimerization rates resembled first-order kinetics in phosphate buffers (pH 7.5 and 9) and in the presence of FBS (pH 3, 7.5 and in Dulbecco's culture media). Epimerization equilibria (48 to 63% ergovaline) were reached within approximately 1 to 19 h. Results from this study indicate that researchers conducting studies with purified ergopeptines should carefully control the storage conditions of solvated ergopeptines and measure isomeric composition under the actual experimental conditions used in experiments. PMID- 12078744 TI - Effects of N-methyl-D,L-aspartate on LH, GH, and testosterone secretion in goat bucks maintained under long or short photoperiods. AB - Photoperiod modulates reproduction in goats. We tested the hypothesis that the excitatory glutamatergic tone is reduced in the photoinhibited goat. The objectives of this study were to determine the effect of photoperiod and glutamatergic stimulation on LH, GH, and testosterone (T) secretion in goat bucks. Eight mature, intact bucks were used in two simultaneous 4 x 4 Latin square designs. Variables were two photoperiod regimens (short day; SD, 10 h light:14 h dark, n = 4; vs long day; LD, 16 h light:8 h dark, n = 4) and four doses of N-methyl-D-L-aspartate (NMA; 0, 1, 2 and 4 mg/kg BW, i.v.). Venous blood was obtained for 2 h before and after NMA injection, followed by GnRH injection and then a final 1 h of sampling. Injection of NMA increased (P < 0.002) LH secretion within 20 min. This increase was sustained for 120 min, but the response was most pronounced in LD goats. The increase in mean LH was associated with a concomitant dose-dependent increase in pulse frequency (P < 0.006). However, NMA treatment had no effect (P > 0.10) on LH pulse amplitude. The release of LH after injection of GnRH was not affected by photoperiod. Exposure of bucks to LD reduced T secretion relative to that of SD bucks (P < 0.01). However, GH secretion was enhanced in LD bucks (P< 0.001). The response of GH to NMA was dependent on photoperiod history. A highly significant immediate and sustained increase (P < 0.001) was observed in LD but not in SD bucks within 10 min. Overall, a dose-dependent increase (P < 0.01) in T secretion was stimulated by NMA in both LD and SD bucks. These results indicate that NMA receptors may be involved in the regulation of LH, GH, and testosterone secretion in the goat. Furthermore, length of day influences GH secretion in the goat and NMA receptor activation had divergent effects on the secretion of this hormone. PMID- 12078745 TI - Influence of rumen protein degradability and supplementation frequency on performance and nitrogen use in ruminants consuming low-quality forage: cow performance and efficiency of nitrogen use in wethers. AB - Two studies were conducted to determine the influence of CP degradability and supplementation frequency (SF) on ruminant performance and N efficiency. Treatments included an unsupplemented control (CON) and degradable intake protein (DIP; 82% of CP) or undegradable intake protein (UIP; 60% of CP) provided daily, every 3 d, or every 6 d. Seven wethers (36+/-1 kg BW) were used in the digestibility study with DIP and UIP treatments formulated to meet CP requirements. Eighty-four Angus x Hereford cows (512+/-42 kg BW) in the last third of gestation were used for the performance study. The DIP treatments were calculated to provide 100% of the DIP requirement and UIP treatments were provided on an isonitrogenous basis compared with DIP. Basal diets consisted of low-quality (5% CP) meadow hay. Forage DMI and N intake by lambs decreased (P < 0.05) linearly as SF decreased. Additionally, DMI, OM intake, N retention, N digestibility, and digested N retained were greater (P < 0.01) for supplemented wethers than for controls with no difference due to crude protein degradability. Nitrogen balance, DMI, and OM intake decreased linearly (P < 0.05) as SF decreased. Plasma urea (PU; mM) was measured over a 6-d period and supplemented lambs had increased (P < 0.01) PU compared with CON. Plasma urea linearly decreased (P < 0.01) as SF decreased. Pre- and postcalving (within 14 d and 24 h of calving, respectively) cow weight and body condition score change were more positive (P < 0.05) for supplemented groups than for controls. Results suggest CP supplements consisting of 20 to 60% UIP can be effectively used by ruminants consuming low-quality forage without adversely affecting N efficiency and animal performance, even when provided as infrequently as once every 6 d. PMID- 12078746 TI - Factors affecting animal performance during the grazing season in a mountain cattle production system. AB - The factors influencing weight changes during the grazing season of Brown Swiss autumn-calving cows and Brown Swiss and Pirenaica spring-calving cows and their calves were studied over an 8-yr period in Spanish mountain conditions. The data set comprised 552 annual production cycles of cows that calved in two consecutive years. The animals grazed on alpine ranges during the summer and on forest pastures in the spring and autumn. They were housed during the winter and fed at different feeding levels (83 to 117% of their energy requirements) throughout the years of study. Weights were recorded every 3 mo and corrected to account for changes of digestive content and fetal growth, using theoretical relationships. Cow weight gains both on forest pastures and high mountain ranges were higher in autumn- than in spring-calving Brown Swiss cows, and therefore also during the whole grazing season (52.1 vs 7.7 kg, respectively, P < 0.001). Therefore, weight at calving and thereafter was significantly higher in autumn- than in spring calving cows, which was associated with better reproductive performance (35.5 vs 49.1 d from calving to first ovulation, P < 0.01). In the spring-calving herd, Pirenaica cows had slightly higher gains than Brown Swiss cows during the grazing period (18.5 vs 7.7 kg, P < 0.001), mainly due to their higher gains on forest pastures, but their reproductive performance was similar (44.5 vs 49.1 d from calving to first ovulation, respectively, not statistically significant). Gains were higher in multiparous than in primiparous cows (31.1 vs 14.1 kg, respectively, P < 0.001), especially in the case of Brown Swiss cows, which were younger at first calving. Gains were affected by year of study (P < 0.001) and previous weight changes during the housing period (r = -0.35 and r = -0.21 in autumn- and spring-calving cows respectively, P < 0.001). In the case of autumn calving cows, performance on pasture was also affected by the stage of pregnancy at housing (r = -0.51, P < 0.001). Growth rates through lactation were higher in autumn- than in spring-born calves (P < 0.001), although the shorter lactation period resulted in lower weight at weaning of the former (P < 0.001). Breed was a significant source of variation in the performance of spring-born calves, weights and gains being higher in Brown Swiss than in Pirenaica calves (P < 0.001). PMID- 12078747 TI - Effect of increasing proportion of supplemental N from urea in prepartum supplements on range beef cow performance and on forage intake and digestibility by steers fed low-quality forage. AB - Four experiments were conducted to evaluate the influence of changing the proportion of supplemental degradable intake protein (DIP) from urea on forage intake, digestion, and performance by beef cattle consuming either low-quality, tallgrass prairie forage (Exp. 1, 2, and 4) or forage sorghum hay (Exp. 3). Experiments 1, 2, and 3 were intended to have four levels of supplemental DIP from urea: 0, 20, 40, and 60%. However, refusal to consume the 60% supplement by cows grazing tallgrass prairie resulted in elimination of this treatment from Exp. 1 and 2. Levels of supplemental DIP from urea in Exp. 4 were 0, 15, 30, and 45%. Supplements contained approximately 30% CP, provided sufficient DIP to maximize digestible OM intake (DOMI) of low-quality forage diets, and were fed to cows during the prepartum period. In Exp. 1, 12 Angus x Hereford steers (average initial BW = 379) were assigned to the 0, 20, and 40% treatments. Forage OM intake, DOMI, OM, and NDF digestion were not affected by urea level. In Exp. 2, 90 pregnant, Angus x Hereford cows (average initial BW = 504 kg and body condition [BC] = 5.0) were assigned to the 0, 20, and 40% treatments. Treatment had little effect on cow BW and BC changes and calf birth weight, ADG, or weaning weight. However, pregnancy rate tended to be lowest (P = 0.13) for the greatest level of urea. In Exp. 3, 120 pregnant, crossbred beef cows (average initial BW = 498 kg and BC = 4.6) were assigned to the 0, 20, 40, and 60% treatments. Prepartum BC change tended (P = 0.08) to be quadratic (least increase for 60% treatment), although BW change was not statistically significant. Treatment effect on calf birth weight was inconsistent (cubic; P = 0.03), but calf ADG and weaning weight were not affected by treatment. Pregnancy rate was not affected by prepartum treatment. In Exp. 4, 132 pregnant, Angus x Hereford cows (average initial BW = 533 and BC = 5.3) were assigned to the 0, 15, 30, and 45% treatments. Prepartum BC loss was greatest (quadratic; P = 0.04) for the high urea (45%) treatment, although BW loss during this period declined linearly (P < 0.01). Prepartum treatment did not affect pregnancy rate, calf birth weight, or ADG. In conclusion, when sufficient DIP was offered to prepartum cows to maximize low-quality forage DOMI, urea could replace between 20 and 40% of the DIP in a high-protein (30%) supplement without significantly altering supplement palatability or cow and calf performance. PMID- 12078748 TI - Restricted suckling of tropical dairy cows by their own calf or other cows' calves. AB - The objective of this study was to compare restricted suckling of tropical cows by their own or another cow's calf with artificial rearing of the calves and no suckling. In Exp. 1, cows were mechanically milked twice daily, after which for 15 min they were either suckled by their own calf (Treatment O) or multiple suckled by other cows' calves (Treatment M) or unsuckled, with the calves reared artificially (Treatment A). Machine milk yield was similar for the three treatments, but in the two suckling treatments the additional milk consumed by the calf increased (P = 0.02) total production (2,682, 2,634, and 2,336 kg/lactation for Treatments O, M, and A, respectively). Machine milk fat concentration was reduced (P = 0.05) by suckling (2.90, 3.07, and 3.20% for Treatments 0, M, and A, respectively), but the milk sampled just before suckling (to represent that taken by the calves) had a high fat concentration (mean 7.9%). Machine milk somatic cell count was also reduced (P = 0.05) by suckling, from 106,000/mL (Treatment A) to 85,000/mL (Treatment M) and 95,000 (Treatment O). Cows suckling their own calf lost more weight and body condition than cows whose calves were reared artificially, with multiple-suckled calves intermediate. Cows suckling their own calf had postpartum interval to first estrus increased (P = 0.01) by 31 d and conception rates to first service of 44% compared to 77% for the other two treatments (P = 0.01). The growth of the suckled calves was compared with that of the artificially reared calves, which were given recommended milk allowances. The artificially reared calves consumed more milk and concentrates, which were available ad libitum to all calves, and gained (P = 0.03) 0.07 kg/d more weight than suckled calves. A second experiment determined that suckling once daily did not reduce reproductive performance compared to artificial rearing. We conclude that suckling cows twice daily increases total milk production but reduces body weight in early lactation. Cows suckling their own calves have reduced reproductive performance compared to those suckling other calves or reared artificially. PMID- 12078749 TI - Energy requirement for maintenance and growth of Nellore bulls and steers fed high-forage diets. AB - Data from three comparative slaughter experiments with individually fed Nellore bulls (n = 31) and steers (n = 66) were utilized to determine their NEm and NEg requirements when fed high-forage diets. The experimental design provided ranges in ME intake, BW, and ADG for the development of regression equations to predict NEm and NEg requirements. The Nellore bulls (Trial 1) were divided into two intake levels (ad libitum and 65% of the ad libitum). The steers (Trials 2 and 3) were allocated to three intake levels (ad libitum and 55 and 70% of the ad libitum). In both trials, there were three slaughter groups within each intake level. The three end points for the bulls were different days on treatment (100, 150, and 190 d and 130, 180, and 200 d, respectively, for older and younger animal subgroups). The steers were slaughtered when animals of the ad libitum treatment reached 400, 440, and 480 kg shrunk BW (SBW) on average for the first, second, and third group, respectively. For all body composition determinations, whole empty body components were weighed, ground, and subsampled for chemical analysis. In each of the trials, initial body composition was determined with equations developed from a baseline slaughter group, using SBW and empty BW (EBW), fat (EBF), and protein (EBP) as variables. The NEm was similar for bulls and steers; NEm averaged 77.2 kcal/ kg0.75 EBW. However, the efficiency of conversion of ME to net energy for maintenance was greater for steers than for bulls (68.8 and 65.6%, respectively), indicating that bulls had a greater ME requirement for maintenance than steers (5.4%; P < 0.05). Our analyses do not support the NRC (2000) conclusion that Nellore, a Bos indicus breed, has a lower net energy requirement for maintenance than Bos taurus breeds. An equation developed with the pooled data to predict retained energy (RE) was similar to the NRC (2000) equation. A second equation was developed to predict RE adjusted for degree of maturity (u): RE = (6.45 - 2.58/u) x EWG x e(0.469) x u), where u = current EBW/final EBW in which final EBW was 365 kg for steers and younger bulls and 456 kg for older bulls at 22% EBF, respectively. PMID- 12078750 TI - Sodium chlorate supplementation reduces E. coli O157:H7 populations in cattle. AB - Cattle are a natural reservoir of the food-borne pathogen Escherichia coli O157:H7. Therefore, strategies that reduce E. coli O157:H7 prior to slaughter will reduce human exposures to this virulent pathogen. When bacteria that can anaerobically respire on nitrate (e.g., E. coli) are exposed to chlorate, they die because the intracellular enzyme nitrate reductase converts nitrate to nitrite, but also co-metabolically reduces chlorate to cytotoxic chlorite. Because chlorate is bactericidal only against nitrate reductase-positive bacteria, it has been suggested that chlorate supplementation be used as a strategy to reduce E. coli O157:H7 populations in cattle prior to harvest. Cattle (n = 8) were fed a feedlot-style high-grain diet experimentally infected with three strains of E. coli O157:H7. Cattle were given access to drinking water supplemented with 2.5 mM KNO3 and 100 mM NaCl (controls; n = 4) or 2.5 mM KNO3 and 100 mM NaClO3 (chlorate-treated; n = 4). Sodium chlorate treatment for 24 h reduced the population of all E. coli O157:H7 strains approximately two logs (10(4) to 10(2)) in the rumen and three logs (10(6) to 10(3)) in the feces. Chlorate treatment reduced total coliforms and generic E. coli from 106 to 10(4) in the rumen and by two logs throughout the rest of the gastrointestinal tract (ileum, cecum, colon, and rectum). Chlorate treatment reduced E. coli O157:H7 counts throughout the intestinal tract but did not alter total culturable anaerobic bacterial counts or the ruminal fermentation pattern. Therefore, it appears that chlorate supplementation is a viable potential strategy to reduce E. coli O157:H7 populations in cattle prior to harvest. PMID- 12078751 TI - Phosphorus requirement of finishing feedlot calves. AB - Dietary P supplied to feedlot cattle is important because an inadequate supply will compromise performance, whereas excess P may harm the environment. However, P requirements of feedlot cattle are not well documented. Therefore, 45 steer calves (265.2+/-16.6 kg) were individually fed to determine the P required for gain and bone integrity over a 204-d finishing period. The basal diet consisted of 33.5% high-moisture corn, 30% brewers grits, 20% corn bran, 7.5% cottonseed hulls, 3% tallow, and 6% supplement. Treatments consisted of 0.16 (no supplemental inorganic P), 0.22, 0.28, 0.34, and 0.40% P (DM basis). Supplemental P was provided by monosodium phosphate top-dressed to the daily feed allotment. Blood was sampled every 56 d to assess P status. At slaughter, phalanx and metacarpal bones were collected from the front leg to determine bone ash and assess P resorption from bone. Dry matter intake and ADG did not change linearly (P > 0.86) or quadratically (P > 0.28) due to P treatment. Feed efficiency was not influenced (P > 0.30) by P treatment and averaged 0.169. Plasma inorganic P averaged across d 56 to 204 responded quadratically, with calves fed 0.16% P having the lowest concentration of plasma inorganic P. However, plasma inorganic P concentration (5.7 mg/dL) for steers fed 0.16% P is generally considered adequate. Total bone ash weight was not influenced by dietary P for phalanx (P = 0.19) or metacarpal bones (P = 0.37). Total P intake ranged from 14.2 to 35.5 g/d. The NRC (1996) recommendation for these calves was 18.7 g/d, assuming 68% absorption. Based on performance results, P requirements for finishing calves is < 0.16% of diet DM or 14.2 g/d. Based on these observations, we suggest that typical grain-based feedlot cattle diets do not require supplementation of inorganic mineral P to meet P requirements. PMID- 12078752 TI - Rapid communication: nucleotide sequence of the bovine elongation factor 1 alpha cDNA1. PMID- 12078753 TI - Rapid communication: nucleotide sequence and physical mapping of the porcine cyclin-dependent kinase inhibitor 3 (CDKN3) gene. PMID- 12078754 TI - Rapid communication: physical and linkage mapping of the porcine calcitonin (CALC) gene. PMID- 12078755 TI - High-grade esophageal dysplasia: long-term survival and quality of life after esophagectomy. AB - BACKGROUND: Esophagectomy for high-grade dysplasia in Barrett's esophagus has been advocated. Although long-term survival data exist, little is known about functional outcome and quality of life in this particular subset of patients. METHODS: The records of all patients who underwent esophageal resection for high grade dysplasia from June 1991 through July 1997 were reviewed. Long-term functional outcome and quality of life were assessed using a two-part written survey. RESULTS: There were 54 patients (48 men, 6 women). Median age was 64 years (range, 36 to 83 years). Ivor Lewis esophagogastrectomy was performed in 34 patients (63%), transhiatal esophagectomy in 10 (18%), extended esophagectomy in 8 (15%), and other in 2 (4%). Invasive carcinoma was found in 19 patients (35%). Five patients (9%) were stage 0, 7 (13%) stage I, 3 (6%) stage IIA, 1 (2%) stage IIB, and 3 patients (6%) stage III. There was one operative death (1.8%). Complications occurred in 31 patients (57%). Median hospitalization was 13 days (range, 11 to 44 days). Follow-up was complete in all patients and ranged from 6 months to 9 years (median, 63 months). Overall 5-year survival was 86% and did not differ significantly from a population matched for age and gender. Five-year survival for patients with only high-grade dysplasia was 96% and 68% for patients with cancer (p = 0.017). Quality of life was measured by the Medical Outcomes Study 36-Item Short-Form Health Survey. For patients with only high-grade dysplasia, the role-physical and role-emotional scores were better than for the control population (p < 0.03). For patients with cancer, the health perception score was worse than for the control population (p < 0.03). Scores measuring physical-function, social function, mental health, bodily pain, and energy/fatigue were similar. CONCLUSIONS: Although perioperative morbidity is significant, surgical resection of high-grade dysplasia in Barrett's esophagus provides excellent long-term survival with acceptable function and quality of life. PMID- 12078756 TI - The influence of perioperative blood transfusion on survival after esophageal resection for carcinoma. AB - BACKGROUND: There is evidence that perioperative blood transfusion may lead to immunosuppression. Our aim was to determine whether blood transfusion influenced survival after esophagectomy for carcinoma. METHODS: The study group comprised 234 consecutive patients (175 men and 59 women) with a mean age of 66 years who underwent esophagectomy for carcinoma by one surgeon between 1988 and 1998. The impact of 41 variables on survival was determined by means of univariate and multivariate analysis. Follow-up was complete (mean follow-up, 19.2 months; standard deviation, 16 months; range, 0 to 129 months). RESULTS: The operative mortality rate was 5.6% (13 deaths). Median operative blood loss was 700 mL (range, 150 to 7,000 mL). One hundred sixty-one patients (68.8%) received a blood transfusion postoperatively (mean transfusion, 2.6 units; range, 0 to 12 units). Overall actuarial 1-year, 3-year, and 5-year survival rates inclusive of operative mortality were 58.1%, 28.5%, and 16.1%, respectively. On univariate analysis, positive lymph nodes, pathological TNM stage, transfusion of more than 3 units of blood, incomplete resection, poor tumor cell differentiation, longer tumor, greater weight loss, male sex, and adenocarcinoma were significant (p < 0.05) negative factors for survival. On Cox proportional hazards regression analysis, after excluding operative mortality, lymph node involvement (p = 0.001), incomplete resection (p = 0.0001), poor tumor cell differentiation (p = 0.04), and transfusion of more than 3 units of blood (p = 0.04) were independent adverse predictors of late survival. CONCLUSIONS: In addition to reaffirming the importance of completeness of resection and nodal involvement, this study demonstrates that blood transfusion (more than 3 units) may have a significant adverse effect on late survival after esophageal resection for carcinoma. Every effort should be made to limit the amount of transfused blood to the absolutely essential requirements. PMID- 12078757 TI - Thoracoscopic and laparoscopic lymph node staging in esophageal cancer: do clinicopathological factors affect the outcome? AB - BACKGROUND: This study was performed to evaluate the pattern of lymphatic metastases found by combined thoracoscopic (TS) and laparoscopic (LS) lymph node staging in esophageal cancer, and ascertain whether clinicopathologic factors may be used to guide the clinical practice of combined TS and LS staging. METHODS: A retrospective study was performed in a series of 76 esophageal cancer patients who had undergone both TS and LS staging before treatment. The correlation of TS and LS lymph node metastases with clinicopathologic factors was analyzed, including the clinical T stage, clinical N stage, tumor location, and histology. RESULTS: Thirty-one patients (40.8%) were found to have lymphatic metastasis by TS and LS staging. Among them, 22 patients had abdominal lymph node metastases, 7 patients had mediastinal lymph node metastases, and 2 patients had both. Patients with advanced T stage (T3 to T4) or adenocarcinoma had a higher frequency of abdominal lymphatic metastases than patients with early T stage (T1 to T2) (39% vs 16%; p = 0.04) or squamous cell carcinoma (39% vs 20%; p = 0.079), respectively. Patients with clinical abdominal N1 stage had a higher incidence of positive laparoscopic finding than patients with clinical abdominal N0 stage (67% vs 23%; p = 0.001). There was no significant correlation between lymphatic metastases and the location of the primary tumor. CONCLUSIONS: Clinicopathologic factors, including the histologic type, the clinical T stage, and abdominal N stage, may affect the outcome of TS and LS lymph node staging in esophageal cancer patients. This clinicopathologic impact may play a role for the selection of candidates for TS and LS staging, and also allows surgeons to focus their attention on the most likely high-yield biopsy targets. PMID- 12078758 TI - Chylothorax complicating pulmonary resection. AB - BACKGROUND: Chylothorax complicating pulmonary resection (CCPR) is infrequent and surgical treatment is for the most part avoided. The purpose of this study is to analyze the clinical and therapeutic characteristics of this complication. METHODS: From March 1981 to June 2001, 26 cases of CCPR (24 men and 2 women; mean age 57 years) were treated in two departments of thoracic surgery. Twenty-five cases complicated lung resection for lung cancer (lobectomy n = 14, bilobectomy n = 3, pneumonectomy n = 8) and 1 case followed lobectomy for a benign lesion. Medical history, location, and characteristics of the chylothorax, lymphography, and clinical evolution after medical or surgical therapy were studied. RESULTS: Medical history was never predictive of CCPR. Chylothorax was right sided in 18 cases and left sided in 8 cases. The total amount of chyle ranged from 1.9 L to 27.9 L per patient with a mean of 7.9 L (pneumonectomy 12.3 L and lobectomy 6.3 L). In 15 patients (pneumonectomy n = 2 and lobectomy n = 13) mean quantity of daily chyle was 0.3 L. All these patients recovered with conservative therapy except for 2 patients who underwent drainage and talc slurry (n = 1) and video assisted lysis of adhesions (n = 1). In the remaining 11 patients (pneumonectomy n = 6 and lobectomy n = 5) mean quantity of daily chyle was 1 L. The chylous leak was seen at lymphography (n = 4), during reoperation (n = 2), or at lymphography and reoperation (n = 3). The location was clearly identified at the level of thoracic duct tributaries in all cases. In 4 postlobectomy cases (4 of 7), surgery was not performed because of the therapeutic usefulness of lymphography. Reoperation was necessary in 6 cases (postpneumonectomy n = 5, postlobectomy n = 1) and consisted of duct ligation (n = 2), leak/suture (n = 3), and fibrin glue (n = 1). CONCLUSIONS: CCPR is rare and appears to respond well to medical treatment owing to the fact that the thoracic duct is generally patent as the leak is due to injury of its tributaries. When surgery is considered, lymphography may help to select cases in which conservative medical therapy should be continued. However, in a small number of cases, usually after pneumonectomy, surgery remains mandatory. PMID- 12078759 TI - Chest wall resections and reconstruction: a 25-year experience. AB - BACKGROUND: Chest wall defects continue to present a complicated treatment scenario for thoracic and reconstructive surgeons. The purpose of this study is to report our 25-year experience with chest wall resections and reconstructions. METHODS: A retrospective review of 200 patients who had chest wall resections from 1975 to 2000 was performed. RESULTS: Patient demographics included tobacco abuse, hypertension, diabetes mellitus, alcohol abuse, coronary artery disease, chronic obstructive pulmonary disease, and human immunodeficiency virus. Surgical indications included lung cancer, breast cancer, chest wall tumors, and severe pectus deformities. Twenty-nine patients had radiation necrosis and 31 patients had lung or chest wall infections. The mean number of ribs resected was 4 +/- 2 ribs. Fifty-six patients underwent sternal resections. In addition 14 patients underwent forequarter amputations. Immediate closure was performed in 195 patients whereas delayed closure was performed in 5 patients. Primary repair without the use of reconstructive techniques was possible in 43 patients. Synthetic chest wall reconstruction was performed using Prolene mesh, Marlex mesh, methyl methacrylate sandwich, Vicryl mesh, and polytetrafluoroethylene. Flaps utilized for soft tissue coverage were free flap (17 patients) and pedicled flap (96 patients). Mean postoperative length of stay was 14 +/- 14 days. Mean intensive care unit stay was 5 +/- 9 days. In-hospital and 30-day survival was 93%. CONCLUSIONS: Chest wall resection with reconstruction utilizing synthetic mesh or local muscle flaps can be performed as a safe, effective one-stage surgical procedure for a variety of major chest wall defects. PMID- 12078760 TI - Predictors and treatment of persistent air leaks. AB - BACKGROUND: Air leaks prolong hospital stay. METHODS: A prospective algorithm was applied to patients. If patients were ready for discharge but still had an air leak, a Heimlich valve was placed and they were discharged. If the leak was still present after 2 weeks, the tube was clamped for a day and removed. RESULTS: There were 669 patients. Factors that predicted a persistent air leak were FEV1% of less than 79% (p = 0.006), history of steroid use (p = 0.002), male gender (p = 0.05), and having a lobectomy (p = 0.01). Types of air leaks on day 1 that eventually required a Heimlich valve were expiratory leaks (p = 0.02), leaks that were an expiratory 4 or more (p < 0.0001), and the presence of a pneumothorax concomitant with an air leak (p < 0.0001). Thirty-three patients were placed on a Heimlich valve, and 6 patients had a pneumothorax or subcutaneous emphysema develop; all patients had an expiratory 5 leak or larger (p < 0.0001). Thirty three patients went home on a valve. Seventeen patients had leaks that resolved by 1 week, 6 by 2 weeks, and the remaining 9 had their tubes removed without problems. CONCLUSIONS: Steroid use, male gender, a large leak, a leak with a pneumothorax, and having a lobectomy are all risk factors for a persistent leak. Discharge on a Heimlich valve is safe and effective for patients with a persistent leak unless the leak is an expiratory 5 or more. Once home on a valve, most air leaks will seal in 2 weeks; if not, chest tubes can be safely removed regardless of the size of the leak or the presence of a pneumothorax. PMID- 12078761 TI - Prognostic significance of neuroendocrine differentiation in adenocarcinoma of the lung. AB - BACKGROUND: The relative incidence of adenocarcinoma of the lung is increasing and some patients with lung carcinoma, detected at an early stage, still develop recurrent disease despite complete resection of the tumor. Recently, neuroendocrine differentiation in large cell carcinoma of the lung has been reported to be of prognostic significance. Therefore, we have evaluated the prognostic significance of neuroendocrine differentiation in adenocarcinoma of the lung. METHODS: A total of 90 resected specimens of adenocarcinoma of the lung measuring 3 cm or less (T1 N0 M0 or T2 N0 M0) were reviewed histologically and immunohistochemical staining was performed to determine the degree of neuroendocrine differentiation. RESULTS: Seven adenocarcinomas exhibited neuroendocrine differentiation in 10% or more of tumor cells. The disease-free survival rate for these patients was significantly lower than that of patients with tumors exhibiting neuroendocrine differentiation in less than 10% of tumor cells or with absent neuroendocrine differentiation (p < 0.0005). Other conventional pathologic factors such as vascular invasion (p < 0.0005), lymphatic invasion (p < 0.05), and pleural involvement (p < 0.05) were also of prognostic significance. In multivariate analysis, the presence of 10% or more neuroendocrine marker-positive tumor cells, vascular invasion, and lymphatic invasion were found to be significantly adverse prognostic factors (p = 0.0162, p = 0.0111, and p = 0.0173, respectively). CONCLUSIONS: Neuroendocrine differentiation of tumor cells is a prognostic factor in lung adenocarcinoma. It is suggested that the identification of neuroendocrine differentiation as well as vascular invasion by tumor in small peripheral adenocarcinoma of the lung may predict the prognosis of these patients. PMID- 12078762 TI - Personal experience in lung cancer sleeve lobectomy and sleeve pneumonectomy. AB - BACKGROUND: Sleeve lobectomy (SL) and tracheal sleeve pneumonectomy (TSP) represent valuable alternative techniques to standard resections in the treatment of benign and malignant conditions of the airway and allow preservation of lung parenchyma. METHODS: Eighty-three sleeve lobectomies and 27 tracheal sleeve pneumonectomies have been performed for nonsmall cell lung cancer in the thoracic department of the University of Milan from 1979 to 1999. There were 46 upper right lobectomies, 11 upper and middle lobectomies, 18 upper left lobectomies, 8 lower left lobectomies, and 27 right pneumonectomies. RESULTS: Mortality rate was 3.6% in SL and 7.4% in TSP. Complications were 10.8% of all SLs and 15% of all TSPs. The overall 5-year survival rate was 43% for SL and 20% for TSP; the 10 year survival rate was 34% and 14%, respectively. There was a highly significant difference in survival between patients with N0 and N1-N2 disease. CONCLUSIONS: Sleeve lobectomy is an appropriate surgical procedure and an alternative to pneumonectomy in patients with limited respiratory reserve whenever the situation permits. Trachael sleeve pneumonectomy is associated with more complications and poor survival. PMID- 12078763 TI - Adenoviral herpes simplex virus thymidine kinase gene therapy in an orthotopic lung cancer model. AB - BACKGROUND: Although adenovirus-mediated herpes simplex virus thymidine kinase (HSVtk) gene therapy is a potential candidate for a novel effective therapy for lung cancer, previous studies have been performed only in a subcutaneous tumor model employing nude mice. We studied therapeutic potentials in correlation with accurate adenoviral gene transduction efficiency in a more clinically relevant orthotopic lung cancer model employing immunocompetent mice. METHODS: To analyze the cytotoxicity of adenoviral HSVtk gene transduction and ganciclovir, a cell proliferation assay was performed in vitro. A survival study was carried out in immunocompetent mice with orthotopic lung cancer, which was generated by intrapulmonary inoculation with syngeneic murine lung cancer cells that had been infected beforehand with each adenoviral vector at the predetermined gene transduction efficiencies. RESULTS: Tumor cells were efficiently killed by infection with adenovirus carrying the HSVtk gene with the addition of ganciclovir in vitro. In the in vivo experiment all control mice died of rapid growth of the primary lung cancer and of metastases to mediastinal lymph nodes within 26 days after tumor inoculation. In contrast 50% and 100% of mice survived more than 40 days after inoculation with adenovirally HSVtk-transfected tumor cells that moderately and highly expressed HSVtk, respectively, when followed by ganciclovir administration. Gene transduction efficiencies were 67%. CONCLUSIONS: Adenovirus-mediated HSVtk gene therapy may be therapeutic for lung cancer when gene transduction efficiencies and sufficient expression levels of HSVtk can be achieved. Moreover, the present findings underscore the importance of the mouse orthotopic lung cancer model for studies of gene therapy. PMID- 12078764 TI - Survival analysis of rats implanted with porous titanium tracheal prosthesis. AB - BACKGROUND: Surgical treatment of a malignancy in the trachea may lead to a long resection that has to be reconstructed with an artificial prosthesis. However, most of the available prostheses encounter inflammatory rejection and mechanical constraint problems. To improve tracheal rehabilitation a porous titanium prosthesis was developed. The aim of this study was to test the biocompatibility of this novel material. METHODS: Seventeen rats had a partial tracheal prosthesis made of porous titanium inserted in the cervical trachea. The histologic analysis of the tissue surrounding the prosthesis was performed in 11 surviving animals after a period of 15 to 41 days. RESULTS: Fibroblast colonization of titanium pores and a ciliary cylindrical epithelial layer developed on the endoluminal side of the prosthesis and the inflammatory reaction was minimal. CONCLUSIONS: The results of this short-term study validate, from surgical and histologic standpoints, the usefulness of a porous titanium tracheal prosthesis. PMID- 12078765 TI - Current incidence of acute neurologic complications after open-heart operations in children. AB - BACKGROUND: Previously, neurologic dysfunction was estimated to complicate 25% or less of pediatric open-heart operations. We sought to determine the current incidence and spectrum of early postoperative neurologic complications. METHODS: We undertook a retrospective review of all patients undergoing open heart operations in 1 year at our institution with the goal to identify all neurologic complications occurring in the early postoperative period. RESULTS: Open-heart operations were performed in 706 children. Sixteen children (2.3%) had neurologic complications develop, including 9 (1.3%) with definite clinical seizures, 1 with suspected seizures and bilateral subdural hemorrhage, 2 with coma after cardiac arrest, 2 with transient mild choreoathetosis, 1 with facial palsy, and 1 with persistent irritability. Causes of seizure were cyclosporin A toxicity posttransplant (4), cerebral ischemia post cardiac arrest (3), and unknown (2). In infants less than 1 year of age, the incidence of seizures was 1.2%. CONCLUSIONS: This review suggests a decrease in acute neurologic morbidity after pediatric open heart operation. Clinical seizures remain the most common complication. Posttransplant, cyclosporin-associated seizures have emerged as an important etiologic category, coincident with an increase in cardiac transplantation in children. PMID- 12078766 TI - Comparable systemic ventricular function in healthy adults and patients with unoperated congenitally corrected transposition using MRI dobutamine stress testing. AB - BACKGROUND: Failure of the systemic right ventricle (RV) often complicates adult survival in unoperated or physiologically repaired congenitally corrected transposition of the great arteries (CCTGA). Healthy controls schematically represent an optimal outcome of anatomic repair, which is increasingly performed to treat CCTGA. Magnetic resonance imaging dobutamine stress testing measures cardiac reserve, and sets to compare the left ventricle of controls with the systemic RV of unoperated and physiologically repaired patients with CCTGA. METHODS: Baseline and stress magnetic resonance imaging (maximum dobutamine dose, 15 microg/kg/min) assessed systemic RV function in 13 minimally or asymptomatic adult patients with CCTGA (unoperated, n = 7; physiologically repaired, n = 6). The left ventricles of 11 healthy age-matched adults served as controls. RESULTS: Baseline and stress end-diastolic volumes similar between the systemic RV of unoperated patients and the left ventricle of controls, as well as base end systolic volumes. Stress ejection fraction was lower in unoperated and physiologically repaired patients (70 +/- 6% and 60 +/- 5%, respectively, vs healthy controls (84 +/- 8%). However, comparable with healthy controls, both subsets of CCTGA patients responded appropriately to dobutamine stress, as illustrated by similar RV stroke volume, heart rate, mean blood pressure, and cardiac index. CONCLUSIONS: Compared with the left ventricles of healthy controls, both patient groups had larger systemic RV volumes, diminished ejection fraction, but an appropriate response to dobutamine stress. Values of unoperated patients are closer to normal than physiologically repaired patients. Magnetic resonance imaging dobutamine may help to define the subgroups of CCTGA patients with favorable anatomy, whereby asymptomatic adult survival could be anticipated without the need for an operation. PMID- 12078767 TI - Change of serum growth factors in infants with isolated ventricular defect undergoing surgical repair. AB - BACKGROUND: Despite increasing clinical use and recent evidence that insulin-like growth factor-1 (IGF-1), insulin-like growth factor binding protein-3 (IGFBP-3), and human growth hormone (hGH) target the heart, the clinical manifestations following the change in the serum growth factors in infants with isolated ventricular septal defect (VSD) undergoing surgical repair have not been clearly defined. METHODS: Twenty normal infants (group I) and 44 consecutive infants with echocardiography established isolated VSD (aged from 3 months to 1 year; body weight from 6.0 +/- 1.8 kg to 8.2 +/- 1.6 kg) were investigated. Among 44 infants with VSD, 20 with shunt fraction, Qp/Qs < or = 1.5 were free of symptoms of congestive heart failure (group II); 24 with shunt fraction, Qp/Qs > or = 2.0 were in congestive heart failure (group IIIa); and 20 of these 24 infants had undergone VSD repair 6 months before their second study (group IIIb). Serum IGF 1, IGFBP-3, and hGH factors were determined by enzyme-linked immunosorbent assay using a monoclonal antibody. RESULTS: The serum levels of IGF-1, IGFBP-3, and hGH factors were 111.9 +/- 2.3 ng/mL, 22.0 +/- 2.3 ng/mL, and 3.6 +/- 0.7 microIU/mL for group I; 63.8 +/- 8.2 ng/mL, 17.1 +/- 1.6 ng/mL, and 4.1 +/- 1.2 microIU/mL for group II; 24.0 +/- 2.6 ng/mL, 9.4 +/- 0.7 ng/mL, and 14.7 +/- 3.5 microIU/mL for group IIIa; 79.4 +/- 12 ng/mL, 20.3 +/- 1.3 ng/mL, and 4.3 +/- 0.7 microIU/mL for group IIIb. In comparison to group I, the decrease in serum levels of IGF-1 and IGFBP-3 in groups II and IIIa were statistically significant (in group II 43% and 32%, p < 0.05; in group IIIa 79% and 37%, p < 0.01). Also the increase in serum level of hGH concentration in group IIIa was significant (increased threefold, p < 0.01). Interestingly, the change in serum levels of IGF-1, IGFBP-3 (decrease), and hGH (increase), returned to the normal range of serum levels after VSD repair in group IIIb. All congestive heart failure symptoms subsided in group IIIb during follow-up. CONCLUSIONS: Improvement in serum levels of IGF-1, IGFBP-3, and hGH were identified in infants with VSD after surgical repair. PMID- 12078768 TI - Coagulation factor abnormalities in patients with single-ventricle physiology immediately prior to the Fontan procedure. AB - BACKGROUND: Coagulation abnormalities have been reported following the Fontan operation and have been attributed to various aspects of Fontan-associated physiology. Using age-matched controls, this study evaluated coagulation abnormalities in children who had undergone a bidirectional Glenn procedure to test the hypothesis that coagulation abnormalities are present before the Fontan operation. METHODS: Coagulation factors were assayed in 38 children (mean age 34.4 +/- 15 months) immediately before the Fontan operation; 37 healthy children (mean age 33 +/- 17 months) were assayed as controls. Concentration of factors II, V, VII, VIII, IX, and X and of antithrombin III, plasminogen, proteins C and S, fibrinogen, serum albumin, and liver enzymes were measured. Normal reference intervals based on the control patients were determined using 95% confidence limits. Patient demographic data, hemodynamic variables, and elapsed time after the Glenn procedure were evaluated as possible predictors of coagulation abnormalities. RESULTS: Concentrations of protein C; factors II, V, VII, and X; plasminogen; and antithrombin III were significantly lower before the Fontan operation compared with age-matched controls (p < 0.01); no specific hemodynamic variables were predictive of a pro- or anticoagulant deficiency. There were significant positive correlations between patients who had abnormally low factor VII, protein S, and protein C levels and a longer interval between the bidirectional Glenn procedure and the Fontan operation (p < 0.001). CONCLUSIONS: Coagulation abnormalities that could predispose patients to increased risk for clotting or bleeding are evident early in the course of staged single-ventricle repair. PMID- 12078769 TI - Valved homograft replacement of aneurysmal pulmonary arteries for severely symptomatic absent pulmonary valve syndrome. AB - BACKGROUND: Patients with absent pulmonary valve syndrome (APVS) with respiratory distress (RD) have previously had a high mortality. In 1990 we adopted a strategy of primary repair including total replacement of the aneurysmal central pulmonary arteries (PAs) for patients with RD. METHODS: Retrospective review was made of 54 consecutive patients with APVS between 1960 and 1998. Median age and weight were 4 months and 4.8 kg. RD was present in 23 patients (10 neonates, 16 required ventilation). Fifteen patients had repair with homograft replacement of the PAs and VSD closure (group 1). Twenty-seven patients had transannular patch with VSD closure with PA-plasty (group 2, n = 21) or without PA plasty (group 3, n = 6). Twelve had miscellaneous procedures (group 4); in 6 the VSD was left open. RESULTS: Operative, 1-, 5-, and 10-year survivals were 83%, 80%, 78%, and 78%, respectively. Risk factors for operative mortality in multivariate analysis were RD (p = 0.04), neonates (p = 0.02), weight less than 3 kg (p = 0.02), open VSD (p = 0.02) and surgery before 1990 (p = 0.04). Since 1990 operative mortality has decreased to 11% (p = 0.04). RD was the only time-related predictor of survival in multivariate analysis (p = 0.004). In patients with RD, survival with homograft was 73% versus 41% with other techniques (p = 0.2). Mean follow-up was 72 +/- 50 months. There were no significant differences in freedom from reintervention rates among the surgical groups (p = 0.08). CONCLUSIONS: Aggressive homograft replacement of the central pulmonary arteries has been associated with improved survival in patients with APVS especially in neonates with severe RD. PMID- 12078770 TI - Repair of Ebstein's anomaly in the symptomatic neonate: an evolution of technique with 7-year follow-up. AB - BACKGROUND: Ebstein's anomaly in the severely symptomatic neonate is usually fatal. Until recently, successful repair has not been reported and various palliative operations have been associated with prohibitive mortality. Recently, we published our initial results with biventricular repair in 3 severely symptomatic neonates. We now update our experience with emphasis on the evolution of our surgical technique and the medium-term follow-up of these patients. METHODS: Since 1994, 8 severely symptomatic neonates and young infants underwent biventricular repair by one surgeon. Six had Ebstein's anomaly and 2 had physiologically similar pathology with severe tricuspid valve dysplasia, cyanosis, and gross cardiomegaly. One Ebstein patient (2 months old) had undergone a Starnes operation elsewhere. Weight of the patients at operation ranged from 2.1 to 6.4 kg (mean 2.7 kg). Five patients had either anatomical (n = 3) or functional (n = 2) pulmonary atresia. Severe (4/4) tricuspid regurgitation was present in all except 1 (Starnes operation), and cardiothoracic ratio exceeded 0.85 in all patients. Echocardiography severity scores were >1.5 in 6 (grade 4/4) and 1.3 in 1 (grade 3/4). Repair consisted of tricuspid valve repair, reduction atrioplasty, relief of right ventricular outflow tract obstruction, partial closure of atrial septal defect, and correction of all associated cardiac defects. Technique of tricuspid valve repair evolved over time: 3 had Danielson type repairs, 3 had DeVega-type repairs, and 2 had complex repairs. RESULTS: One patient died in hospital: a 2.1 kg patient with tricuspid dysplasia, anatomical pulmonary atresia, and hypoplastic pulmonary arteries. The other 7 patients are all in functional class I and in sinus rhythm. Although 3 patients had symptomatic tachyarrhythmias before surgery, no child has experienced SVT after discharge. At recent echocardiography 4 patients had mild tricuspid regurgitation, and 2 had mild-moderate (2/4) tricuspid regurgitation. Three patients are now 7 years old, 2 are almost 2 years old, and the remaining 2 patients are 1 year old. CONCLUSIONS: Surgical repair of the severely symptomatic neonate with Ebstein's anomaly is feasible and safe. The repair appears durable and with good medium-term outcome. PMID- 12078771 TI - Right ventricular dysfunction and pulmonary valve replacement after correction of tetralogy of Fallot. AB - BACKGROUND: Correction of tetralogy of Fallot often leads to pulmonary regurgitation, sometimes warranting pulmonary valve replacement, for which indications and timing to achieve optimal results are not yet clear. This retrospective study describes follow-up and reinterventions in our tetralogy of Fallot population. METHODS: Review of all consecutive patients operated on for tetralogy of Fallot between 1977 and 2000 was conducted. Included are date and type of repair, Doppler echocardiography (two-dimensional echocardiography), electrocardiographs, reoperations, and physical condition. RESULTS: Total repair was performed in 171 patients at a mean age 1.9 +/- 2.5 years, follow-up time counted 9.6 +/- 7.0 years. Right ventriculotomy was used in 92%, and transatrial ventricular septal defect closure was used in 8%; 74% received a transannular outflow patch. Twenty-year survival was 91%. Last follow-up electrocardiographs showed right bundle branch block in 67% and serious arrhythmias in 11%. Two dimensional echocardiography demonstrated severe pulmonary insufficiency and dilated right ventricle in 31% and 38%, respectively, increasing with postrepair age (p < 0.001). Poor clinical condition (New York Heart Association class II+) and echocardiographic proof of right atrial dilatation (p = 0.012) and arrhythmias (p = 0.03) were significantly associated. Furthermore, the influence of residual hemodynamic lesions, such as a remaining ventricular septal defect or pulmonary stenosis, or right ventricular dilatation was important (p = 0.04). Reintervention was necessary in 32 patients (19%; 10-year freedom, 83%), including angioplasty for residual stenosis and pulmonary valve replacement. At a mean age of 9.2 years after correction, 14 patients received a homograft, and 2 patients received a heterograft. In 7 patients the right ventricle returned to normal dimensions and symptoms disappeared. The incidence of right ventricular dilatation was considerably higher (p = 0.020) in patients with a transannular patch; the transatrial approach showed the opposite (p = 0.03), and patients presented with lower QRS duration (p = 0.007), although no difference could be found between survival after both surgical techniques. Effects of early timing (correction < 6 months) on right ventricular dysfunction could not be established. CONCLUSIONS: Severe right ventricular dilatation and pulmonary regurgitation secondary to outflow tract repair in tetralogy of Fallot are frequently occurring sequelae developing slowly over time. Indications for pulmonary valve replacement remain controversial because echocardiographic findings or arrhythmias are not always accompanied by deterioration of clinical condition. However, right atrial dilatation and additional hemodynamic lesions demand increased vigilance. Transatrial repair is associated with a favorable outcome. PMID- 12078772 TI - One hundred pulmonary valve replacements in children after relief of right ventricular outflow tract obstruction. AB - BACKGROUND: Surgical repair of obstructive lesions of the right ventricular outflow tract (RVOT) in children commonly creates pulmonary valve incompetence that may eventually require pulmonary valve replacement (PVR). We reviewed our experience with PVR late after RVOT reconstruction. METHODS: We performed 100 PVRs in 93 children 1.1 months to 22.4 years (median 8) after RVOT reconstruction. Children with right ventricular to pulmonary artery conduits and primary PVRs were excluded. Age at PVR was 4.5 months to 27.9 years (median 9.5 years). Initial diagnosis was tetralogy of Fallot and variants, 62; critical pulmonary stenosis, 15; pulmonary atresia with intact ventricular septum, 7; and others, 9. Eleven patients had a redo PVR. A total of 62 PVRs were homografts; 38 were porcine valves. RESULTS: There was one early death. On follow-up of 5 months to 12.4 years (mean 4.9 years) there were no late deaths although 1 child underwent cardiac transplantation. Actuarial freedom from redo PVR at 8 years was 100% for porcine valves but 70% for homograft valves (p = 0.17). For children younger than 3 years at PVR, freedom from reoperation was 76% at 1 year and 39% at 8 years compared with freedom from redo PVR at 8 years of 100% for children older than 3 years. On latest echocardiogram 97% of porcine valves had mild or no pulmonary regurgitation compared with 72% of homograft valves. CONCLUSIONS: PVR after RVOT reconstruction can be performed with low risk. Porcine valves may be superior to homograft valves although this advantage may be due to older age at time of PVR. PMID- 12078773 TI - Partial replacement of the tricuspid valve by mitral homografts in acute endocarditis. AB - BACKGROUND: Seven patients with acute tricuspid endocarditis underwent partial replacement of the tricuspid valve using mitral homograft tissue. Valve function was evaluated at midterm. METHODS: Operative indications were uncontrolled sepsis in all cases associated with heart failure symptoms in 3 patients and septic pulmonary emboli in 2 patients. These patients were referred to our institution after a course of antibiotic treatment ranging from 7 to 12 weeks. Lesions found at the level of the anterior leaflet of the tricuspid valve were vegetations and rupture of more than half of the marginal cords in all patients. Vegetations were also found on the posterior leaflet in 5 patients. In all instances the septal leaflet was free of lesions. The aortic valve was involved in 4 patients and the pulmonary valve in 1 patient. All patients underwent resection of the anterior and posterior leaflets of the tricuspid valve with their corresponding papillary muscles leaving the septal leaflet in place. Replacement of the tricuspid valve was performed through a right longitudinal atrial access, using the anterior leaflet of a mitral homograft alone in 3 patients and the anterior leaflet with part of posterior leaflet in 4 patients. Associated procedures included aortic valve replacement by a homograft (n = 4) and pulmonary valve reconstruction (n = 1). RESULTS: No hospital deaths are reported. One late death, at 16 months, is reported after reoperation due to recurrent aortic valve endocarditis. At midterm (mean follow-up, 30 months) patients had excellent functional status and normal valvular function during echocardiographic studies. CONCLUSIONS: We conclude that when the degree of tricuspid valve destruction prevents repair, partial homograft replacement can be used as an extension of the already existing reconstructive techniques, with excellent functional results. PMID- 12078775 TI - Eight-year results of aortic root replacement with the freestyle stentless porcine aortic root bioprosthesis. AB - BACKGROUND: Stentless porcine aortic valves offer several advantages over traditional valves. Among these are superior hemodynamics, laminar flow patterns, lack of need for anticoagulation, and perhaps improved durability. METHODS: One hundred four patients were operated on from September 17, 1992, to October 31, 1997, as part of a multicenter worldwide investigation of the Medtronic Freestyle stentless porcine bioprosthesis. All patients received a total aortic root replacement. The patients were evaluated postoperatively at discharge, at 3 to 6 months, and yearly by clinical examination and color flow Doppler echocardiography. RESULTS: Operative mortality was 3.9%. No patient experienced structural valve deterioration, nonstructural deterioration, perivalvular leak, or unacceptable hemodynamic performance. At 8 years, survival was 59.8%. Freedom from thromboembolic complications was 83.3%. Freedom from postoperative endocarditis was 96.9%. Freedom from reoperation was 100%. Mean systolic gradients did not change over the time period studied. They were 6.4 +/- 3.8 mm Hg at 1 year and 6.7 +/- 2.6 mm Hg at 8 years. Correspondingly, effective orifice area was 1.9 +/- 0.7 cm2 at 1 year and 1.8 +/- 0.8 cm2 at 8 years. The incidence of any aortic insufficiency also did not change over the length of follow-up. At 1 year, 98% of patients had no or trivial aortic insufficiency and 2% had mild aortic insufficiency. At 8 years, 100% of patients evaluated were free of any aortic insufficiency. CONCLUSIONS: The Medtronic Freestyle aortic root bioprosthesis can be used safely to replace the aortic root or aortic valve for aortic valve and aortic root pathology. Total root replacement allows optimal hemodynamic performance with no significant aortic regurgitation. Results up to 8 years show excellent survival and no signs of degeneration. Further follow-up is still needed to determine valve durability. PMID- 12078774 TI - Endocarditis after mitral valve repair. AB - BACKGROUND: Native valve endocarditis is frequently managed with antibiotics alone, but prosthetic valve endocarditis usually requires an early operation. What is the best treatment of endocarditis after mitral valve repair? METHODS: From 1986 to 2000, 22 patients were treated for endocarditis affecting a previously repaired mitral valve. Causes of mitral valve dysfunction that led to repair were degenerative (11 patients), ischemic (5 patients), endocarditic (3 patients), rheumatic (2 patients), and functional (1 patient). Endocarditis was active in 21 patients and healed in 1. Interval from initial mitral valve repair to onset of endocarditis ranged from 1 week to 10.3 years (median, 6 months). Pathology included leaflet vegetation (15), annuloplasty vegetation (4), leaflet perforation (5), and abscess (3). Mean follow-up was 3.9 +/- 3.3 years. RESULTS: Fifteen patients underwent repeat mitral valve operations with freedom from mitral valve reoperation of 65%, 41%, and 26% at 30 days, 1 year, and 5 years after onset of endocarditis. After a high early hazard, risk of reoperation fell to 10.8% per year. Seven patients, all with a leaflet vegetation, were treated with antibiotics alone. Antibiotics eradicated infection in all; however all had mitral regurgitation 2+ to 4+. Survival was 96%, 74%, and 68% at 30 days, 1 year, and 5 years. Endocarditis recurred in 1 patient (92% free of event). CONCLUSIONS: Most patients that have endocarditis develop after mitral valve repair require reoperation. However if infection is limited to a leaflet, early reoperation may be unnecessary because antibiotics alone can eradicate infection. PMID- 12078776 TI - Patient prosthesis mismatch is rare after aortic valve replacement: valve size may be irrelevant. AB - BACKGROUND: Although small valve size and patient-prosthesis mismatch are both considered to decrease long-term survival, little direct evidence exists to support this hypothesis. METHODS: To assess the prevalence of patient-prosthesis mismatch and the influence of small valve size on survival, we prospectively studied 1,129 consecutive patients undergoing aortic valve replacement between 1990 and 2000. Mean and peak gradients and indexed effective orifice area were measured by transthoracic echocardiography postoperatively (3 months to 10 years). Abnormal postoperative gradients were defined as those patients with mean or peak gradient above the 90th percentile (mean gradient > or = 21 or peak gradient > or = 38 mm Hg). Patient-prosthesis mismatch was defined as those patients with indexed effective orifice area below the 10th percentile (< 0.60 cm2/m2). RESULTS: A multivariable analysis identified internal diameter of the implanted valve as the only independent predictor of abnormal gradients postoperatively. However, there was no significant difference in actuarial survival between normal and abnormal gradient groups (7 years: 91.2% +/- 1.5% versus 95.0% +/- 2.2%; p = 0.48). Freedom from New York Heart Association class III or IV (7 years: 74.5% +/- 3.1% versus 74.6% +/- 6.2%; p = 0.66) and left ventricular mass index were not different between normal and abnormal gradient groups. Patients with and without patient-prosthesis mismatch were similar with respect to postoperative left ventricular mass index, 7-year survival (95.1% +/- 1.3% versus 94.7% +/- 3.0%; p = 0.54), and 7-year freedom from New York Heart Association class III or IV (79.3% +/- 6.6% versus 74.5% +/- 2.5%; p = 0.40). In patients with patient-prosthesis mismatch and abnormal gradients, the majority had prosthesis dysfunction owing to degeneration. CONCLUSIONS: Severe patient prosthesis mismatch is rare after aortic valve replacement. Patient-prosthesis mismatch, abnormal gradient, and the size of valve implanted do not influence left ventricular mass index or intermediate-term survival. PMID- 12078777 TI - Preliminary experience with the St. Jude Medical Regent mechanical heart valve in the aortic position: early in vivo hemodynamic results. AB - BACKGROUND: The St. Jude Medical Regent is a new generation mechanical aortic valve. METHODS: Between March 2000 and July 2001, this valve was implanted in the aortic position in 40 patients (21 men; mean age 59.1 +/- 9.0 years). Preoperatively, 24 patients (60%) were in New York Heart Association functional class III or IV. Eighteen patients (45%) underwent associated procedures. Mean valve size was 21.4 +/- 2.4 mm. The mean duration of follow-up was 8.5 +/- 4.5 months (range, 1 to 16 months). RESULTS: There were no operative deaths. Early complications included one reoperation for bleeding and one transient low output syndrome. Valve replacement was followed by a significant reduction in mean and peak transaortic gradients over time (p < 0.001) and analysis of variance failed to demonstrate statistical differences between valve size over time (p = not significant). A significant reduction in left ventricular hypertrophy occurred over time (p = 0.01) in all valve sizes (p = not significant between groups): baseline left ventricular mass index was 194 g/cm2; it reduced by 22 g/cm2 (p = 0.006) at discharge. Left ventricular mass index decreased from 172 +/- 55 g/cm2 to 156 +/- 44 g/cm2 (p = 0.03) from discharge to 2 months. Further reductions were not significant. Relative wall thickness decreased from 0.57 +/- 0.13 preoperatively to 0.42 +/- 0.06 at discharge (p = 0.001), and again at 2 months ( 0.2; p = not significant), and at 1 year (-0.02; p = not significant). CONCLUSIONS: The early experience with the St. Jude Medical Regent valve has been satisfactory. PMID- 12078778 TI - Aortic arch repair with right brachial artery perfusion. AB - BACKGROUND: To determine the effectiveness of unilateral selective cerebral perfusion for aortic arch repair and to discuss possible modifications to enhance technical simplicity. METHODS: In the period between January 1996 and April 2001, 104 patients underwent aortic arch repair with the use of right brachial artery low flow (8 to 10 mL/kg per minute) antegrade selective cerebral perfusion under moderate hypothermia (26 degrees C). Mean patient age was 52 +/- 12 years. Sixty four patients presented with Stanford type A aortic dissection, including 12 with acute dissection; 38 patients had aneurysmal dilatation of the ascending aorta and aortic arch; and 2 patients had isolated arch aneurysm. Ascending and partial arch replacement was performed in 50 patients; ascending and total arch replacement in 33 patients; ascending and descending arch replacement in 19 patients; and isolated arch replacement in 2 patients. RESULTS: Mean antegrade cerebral perfusion time was 39 +/- 22 minutes. One patient with acute proximal dissection died because of cerebral complications. One other patient developed right hemiparesis, which resolved during the second postoperative month without sequela. Other than these 2 cases (1.9%), no other neurologic event was observed. CONCLUSIONS: The technique of low flow antegrade selective cerebral perfusion through the right brachial artery may be used for a vast majority of aortic aneurysms and dissections requiring arch repair. This technique does not necessitate deep hypothermia, requires shorter cardiopulmonary bypass and operation times, has the advantage of simplicity, provides optimal vascular repair without time restraints and, in terms of clinical results, is as safe as other techniques for cerebral protection. PMID- 12078779 TI - Surgical treatment of acute type A dissection: is rupture a risk factor? AB - BACKGROUND: The purpose of this study was to evaluate the significance of aortic rupture on clinical outcome in patients after aortic repair for acute type A dissection. METHODS: One hundred and twenty patients underwent aortic operations with resection of the intimal tear and open distal anastomosis. Median age was 60 years (range 16 to 87); 78 were male. Thirty-six patients had only ascending aortic replacement, 82 had hemiarch repair, and 2 had the entire arch replaced. Retrograde cerebral perfusion was utilized in 66 patients (53%). Rupture defined as free blood in the pericardial space was present in 60 patients (50%). Univariate and multivariate analyses were performed to assess the risk factors for mortality and neurologic dysfunction. RESULTS: Overall hospital mortality rate was 24.2% +/- 4.0% (+/- 70% confidence level) but did not differ between patients with aortic rupture or without (p = 0.83). The incidence of permanent neurologic dysfunction was 9.4% overall, 10.5% with rupture and 8.3% without rupture (p = 0.75). Multivariate analysis revealed absence of retrograde cerebral perfusion and any postoperative complication as statistically significant indicators for in-hospital mortality (p < 0.05). Overall 1- and 5-year survival was 85.3% and 33.7%; among discharged patients, survival in the nonruptured group was 89% and 37%, versus 81% and 31% in the ruptured group (p = 0.01). CONCLUSIONS: Aortic rupture at the time of surgery does not increase the risk of hospital mortality or permanent neurologic complications in patients with acute type A dissections. However, aortic rupture at the time of surgery does influence long-term survival. PMID- 12078780 TI - Coronary hybrid revascularization from January 1997 to January 2001: a clinical follow-up. AB - BACKGROUND: Hybrid revascularization (HyR), combining minimally invasive left internal mammary artery (LIMA) bypass grafting to the left anterior descending coronary artery (LAD) and catheter interventional treatment of the remaining coronary lesions, avoids the disadvantages associated with cardiopulmonary bypass (CPB). We investigated the clinical follow-up of 57 patients with multivessel disease undergoing this procedure in the last 4 years. METHODS: Between January 1997 and January 2001, 57 consecutive patients (41 men and 16 women, aged 65.7 +/ 7.9 years) with coronary artery disease (two-vessel, n = 34; three-vessel, n = 23) were treated with off-pump LIMA-to-LAD bypass combined with balloon angioplasty and stenting of the remaining significantly obstructed (> 50%) coronary vessels. Clinical follow-up data included a early postoperative and a 6 month control angiography and a patient interview in January 2001. RESULTS: All patients underwent LIMA-to-LAD bypass-grafting and balloon angioplasty in 72 coronary lesions without procedural-related complications. However, one early LIMA bypass occlusion was documented during coronary angiography. Postoperatively no deterioration of preexistent organ dysfunction was observed in any patient. The mean follow-up was 100.7 +/- 37.9 weeks in 55 of 57 patients (97%). Control angiography 6 months after HyR (n = 34) revealed a patent LIMA bypass in 33 patients and 8 in-stent restenoses (> 50%) in the coronary arteries that were treated interventionally by re-PTCA (n = 6) or by conventional CABG (n = 1). In 1 patient medical treatment resulted in significant reduction of angina so no further intervention was considered necessary. After HyR 1 patient died 18 months later of an intracerebral hemorrhage. All other patients are alive and doing well. CONCLUSIONS: Our results indicate that in selected patients with multivessel disease including left main stem stenosis HyR is an effective and secure procedure with excellent early and good midterm results. Especially elderly patients with severe concomitant diseases appear to benefit from this approach by avoiding CPB. PMID- 12078781 TI - Right internal mammary artery and radial artery composite in situ pedicle graft in coronary artery bypass grafting. AB - BACKGROUND: The use of two internal mammary artery grafts in coronary artery bypass grafting has been associated with decreased risks of death, reoperation, and angioplasty. However, bilateral internal mammary artery takedown is associated with higher incidence of sternal wound infection, particularly in people with diabetes and in elderly and obese patients. This study was conducted to explore the feasibility of using right internal mammary artery (RIMA) and radial artery (RA) as a composite graft while preserving the distal two thirds of the RIMA to leave the sternal blood supply intact. METHODS: Eighteen patients underwent coronary artery bypass grafting using proximal RIMA and RA composite graft as one of the bypass conduits. The distal two thirds of the RIMA was left intact to preserve sternal blood supply. The graft-free flows of the RIMA and RA composite graft and of the left internal mammary artery graft and the length of the composite graft had been measured. The graft patency and the flow in the distal part of the unharvested RIMA was evaluated postoperatively 2 weeks after the procedure. In 6 of these patients the graft patency was evaluated by selective angiography. RESULTS: There was no hospital mortality or incidence of perioperative myocardial infarction. None of the patients needed intraaortic balloon pump support postoperatively. There was no sternal wound infection. The vessels grafted were distal right coronary artery (n = 7), posterior descending artery (n = 8), obtuse marginal branches (n = 3), and posterolateral ventricular branch (n = 1); 1 patient received the composite graft as a sequential graft to the posterior descending artery and posterolateral left ventricular branches. The mean graft-free flow of the RIMA and RA composite graft was 98.06 +/- 16.93 mL/min compared to left internal mammary artery flows of 55.80 +/- 8.99 mL/min. All 16 patients who had a good echo window showed patent grafts when evaluated by two-dimensional echocardiography and color Doppler echocardiography. All of the 6 patients in whom the angiogram was repeated postoperatively showed patent RIMA and RA grafts. CONCLUSIONS: Myocardial revascularization using proximal RIMA and RA in situ pedicle graft was safe in patients with diabetes and in obese and chronic obstructive pulmonary disease patients. This graft was useful to revascularize posterior descending artery, posterolateral ventricular branches of right coronary artery, and obtuse marginal branches where a left internal mammary artery and RA composite graft cannot be used because of technical reasons. Its usage was not associated with sternal wound infection. PMID- 12078782 TI - Effects of cerivastatin on vascular function of human radial and left internal thoracic arteries. AB - BACKGROUND: Statins may enhance vascular function independently of effects on cholesterol. This study investigated the ability of statins to modulate the vascular recovery of arteries used as coronary bypass grafts. METHODS: Specimens of radial artery and left internal thoracic artery were obtained during coronary artery bypass grafting. The specimens were divided into vascular rings, which were incubated in the absence or presence of cerivastatin (10(-6) mol/L) for either 2 or 24 hours. Using an organ bath technique, endothelial function was examined using acetylcholine (10(-9) to 10(-5) mol/L) after contraction by 3x10( 8) mol/L of endothelin-1. RESULTS: Time-related endothelial dysfunction was shown in the control group of radial artery but not in the cerivastatin group: maximal endothelium-dependent vasodilation in the control and cerivastatin groups were 56.8% +/- 10.2% and 65.9% +/- 10.1% at 2 hours and 39.4% +/- 4.7% and 68.4% +/- 5.0% (p < 0.01, vs control) at 24 hours, respectively. On the other hand, in the left internal thoracic artery, those in the control and cerivastatin groups were 38.3% +/- 8.2% and 45.0% +/- 5.5% at 2 hours and 38.1% +/- 8.2% and 56.5% +/- 8.8% at 24 hours, respectively (NS). CONCLUSIONS: In radial artery, cerivastatin significantly preserved endothelium-dependent vasodilation, which diminished with time in the control group. This could have very important implications in the clinical practice of coronary artery bypass grafting. PMID- 12078783 TI - Evaluation of the effectiveness of off-pump coronary artery bypass grafting in high-risk patients: an observational study. AB - BACKGROUND: Coronary artery bypass grafting in high-risk patients carries substantial morbidity. We compared the effectiveness of off-pump revascularization with that of conventional coronary artery bypass grafting using cardiopulmonary bypass and cardioplegic arrest in consecutive high-risk patients. METHODS: From April 1996 to December 2000, clinical data for consecutive patients undergoing coronary artery revascularization were prospectively entered into a database. Data were extracted for all patients considered to be high risk, defined as the presence of one or more of ten adverse prognostic factors. Hospital mortality and early morbidity were compared between two groups of patients, the on-pump and off-pump groups. RESULTS: The study group comprised 1,570 consecutive high-risk patients, 332 (21.1%) of whom underwent an off-pump operation. Patients in the on-pump group had fewer high-risk factors and lower Parsonnet scores and were less likely to be 75 years of age or older, to have peripheral vascular disease or hypercholesterolemia, or to have sustained a previous transient ischemic attack. However, they were more likely to be assigned to a higher Canadian Cardiovascular Society class and had more extensive coronary artery disease and were more likely to have unstable angina, to require urgent or emergency operations, and to receive more grafts than those undergoing off-pump procedures. Unadjusted odds ratios for intensive care unit or high-dependency unit stay, total length of stay, blood loss of more than 1,000 mL, postoperative hemoglobin and transfusion requirement all showed a highly significant benefit for the off-pump group (p < or = 0.005; odds ratios, 0.33 to 0.65). After adjustment for prognostic variables, odds ratios remained essentially unaltered (adjusted odds ratio estimates 0.36 to p < 0.05) except for blood loss of more than 1,000 mL (adjusted odds ratio estimate, 0.82; p = 0.22). Sensitivity analyses confirmed the robustness of these findings. CONCLUSIONS: Off-pump coronary artery bypass grafting is safe, effective, and associated with reduced morbidity in high-risk patients compared with conventional coronary artery revascularization. PMID- 12078784 TI - Hemodynamic collapse during off-pump coronary artery bypass grafting. AB - BACKGROUND: The causes of hemodynamic collapse during off-pump coronary artery bypass (OPCAB) remain scarcely defined. We present an analysis of 23 cases of sustained hemodynamic collapse during elective off-pump CABG. METHODS: During a 54-month period, we performed 1420 elective OPCAB procedures through a sternotomy, constituting 71.2% of the total CABG procedures performed. Twenty three patients (1.6%) experienced hemodynamic collapse intra-operatively requiring immediate cardiopulmonary bypass. Preoperative characteristics, intraoperative data, and postoperative outcome were retrospectively reviewed in each patient. RESULTS: In all cases, improvements in intra-operative technique and/or judgment could be made retrospectively. Twenty (20/23) of these patients had an uneventful postoperative course and three (3/23) patients had an unstable course with two deaths. CONCLUSIONS: The causes of hemodynamic collapse during elective OPCAB were ischemic, mechanical, or a combination of both. A detailed review of our five and a half year experience has revealed a number of suggestions for improving the conduct of the operation. PMID- 12078785 TI - Skeletonized radial artery grafting: improved angiographic results. AB - BACKGROUND: The radial artery has been used for coronary artery bypass grafting (CABG) but its early angiographic results were relatively inferior to that of the internal mammary artery, most likely due to spasm of the graft. To avoid vasospasm we harvested the radial artery using a skeletonized technique and spasm was completely reversed before use. The graft patency of the skeletonized radial artery was compared with the radial artery graft harvested as a pedicle. METHODS: A total of 112 patients underwent isolated CABG using a pedicled radial artery between September 1, 1999, and August 31, 2000 (group P), and a total of 131 patients with a skeletonized radial artery between September 1, 2000, and August 31, 2001 (group S). An ultrasonic scalpel (Harmonic Scalpel; Ethicon EndoSurgery, Cincinnati, OH) was used for skeletonization and removing satellite veins and surrounding tissue. CABG was performed by the standard technique. Perioperative results were prospectively collected and compared between the two groups. Early angiographic results performed within 3 months were also compared. RESULTS: There were two hospital deaths in group S. Major complications were observed in 11 (8.4%) in group S and 3 (2.7%) in group P (p = not significant [NS]). None were related to the radial artery graft. Angiography was obtained in 96 patients of group S and 76 patients in group P and revealed that the stenosis free graft patency rate of group S (138 of 143, 96.5%) was superior to that of group P (73 of 86, 84.9%) with p < 0.005. CONCLUSIONS: Skeletonization of the radial artery with the ultrasonic scalpel is safe and contributes to reducing the incidence of early graft stenosis. PMID- 12078786 TI - Factors influencing results and outcome after transmyocardial laser revascularization. AB - BACKGROUND: Transmyocardial laser revascularization (TMR) has been increasingly used during the recent past for patients with diffuse coronary artery disease. Because this operation is associated with significant morbidity and mortality, it is important to select patients for TMR who are likely to benefit from the procedure. METHODS: We performed an univariate logistic regression analysis of 20 factors on the benefits and outcomes of 134 patients who underwent isolated TMR at our institution between November 1994 and May 2000. RESULTS: Responders and nonresponders differed significantly with regard to the incidence of diabetes mellitus. For diabetic patients the chance of profiting from the TMR operation was only 43% of that of nondiabetic patients (odds ratio = 0.43 [0.20 to 0.92]). Furthermore, patients with a preoperative body mass index of less than 25 had a threefold increase in the probability of death during the first year after TMR as compared to patients whose body mass index was 25 or more (odds ratio = 2.97 [1.05 to 8.40]). The incidence of diabetes was also slightly but not significantly different between 1-year survivors and nonsurvivors. CONCLUSIONS: In diabetic patients we recommend caution in selecting therapeutic TMR because outcomes are less satisfactory than in nondiabetics. Furthermore, patients with a body mass index below 25 have a significantly higher risk for death during the first postoperative year. Studies based on larger patient populations should follow. PMID- 12078787 TI - Laser-assist during extraction of chronically implanted pacemaker and defibrillator leads. AB - BACKGROUND: Extraction of chronically implanted pacing and defibrillator leads has historically been difficult, occasionally requiring open surgical procedures. The purpose of this study was to evaluate the efficacy, safety, and potential need for percutaneous laser-assisted sheath techniques for extraction of chronically implanted leads. METHODS: From January 1999 to August 2001, 128 consecutive patients underwent extraction of 229 leads (138 pacing, 91 defibrillator) in the operating room 61 +/- 44 (mean +/- standard deviation) months after implantation. Common indications included erosion or pocket infection (41%), lead dysfunction (30%), and sepsis (13%). RESULTS: Laser techniques were used for 56% +/- 4% (104 of 186) of long-term (implanted for more than 1 year) leads, compared with only 16% +/- 6% (7 of 43) of short-term (implanted for less than 1 year) leads (p < 0.001). For infected leads, laser was used in 53% +/- 5% (49 of 92) with erosion or pocket infections, compared with only 3% +/- 4% (1 of 29) with sepsis (p < 0.001). Extraction was complete in 88%, near complete (retained tip) in 10%, and incomplete in 2%. Two patients required a later percutaneous femoral venous approach to remove mobile retained segments, but no patients required cardiac surgery for extraction. Complications included sternotomy for subclavian vein injury (1), chest tube for caval perforation (1), innominate vein thrombosis (1), and partial clavicle removal for subclavian vein repair (2). There were no procedure-related deaths. CONCLUSIONS: Laser-assisted lead extraction is safe, but it is best performed in the operating room; it should be available for long-term leads, except when they are grossly infected, producing sepsis. Laser techniques have essentially eliminated the need for open surgical removal of retained leads. PMID- 12078788 TI - Aprotinin in coronary operation with cardiopulmonary bypass: does "low-dose" aprotinin inhibit the inflammatory response? AB - BACKGROUND: Cardiopulmonary bypass induces a systemic inflammatory response. Aprotinin, a nonspecific proteinase inhibitor is known to improve postoperative hemostasis and may modify the inflammatory reaction. This study evaluates the effects of low-dose aprotinin on inflammatory markers in patients scheduled for elective coronary artery bypass grafting. METHODS: Patients were prospectively randomized into two groups: the control group (C) (n = 14) and the low-dose aprotinin group (A) (n = 15) with (2 x 10(6) KIU = 280 mg) aprotinin added to the pump prime. Cytokine response (interleukin-6, soluble TNF II receptor), terminal complement production (SC5b-9), and neutrophil activation (lactoferrin) were assessed up to 6 hours postoperatively. Clinical data and hemostatic factors including fibrinopeptide A, thrombin-antithrombin complex, D-dimer, and plasmin/alpha2-antiplasmin were investigated. RESULTS: In both study groups, a significant increase of all inflammatory markers was seen (IL-6, sTNF-IIR, SC5b 9, lactoferrin), p less than 0.001. Peak levels of complement production occurred after protamine administration, whereas cytokine increases were more pronounced postoperatively with marked elevation up to 6 hours. The markers did not differ significantly between groups throughout the study period (p > 0.05 at each time of determination). However, after protamine administration reduced fibrinolysis (D-dimer, plasmin/alpha2-antiplasmin) was detected in group A. Measurements for coagulation (fibrinopeptide A, thrombin-antithrombin complex) were not significantly influenced by aprotinin. The total amount of blood loss during the first 24 hours was significantly reduced in group A (p < 0.02). CONCLUSIONS: Low dose aprotinin added to the pump prime does not inhibit the inflammatory response caused by cardiopulmonary bypass, but improves postoperative hemostasis. A potential effect of high-dose aprotinin on inflammatory markers remains to be elucidated. PMID- 12078789 TI - Lactic acidosis after cardiac surgery is associated with polymorphisms in tumor necrosis factor and interleukin 10 genes. AB - BACKGROUND: Lactic acidosis after cardiac surgery is a manifestation of excess cytokine production. Cytokine-related genetic polymorphisms account for variability in cytokine response and may predispose to the development of lactic acidosis after cardiac surgery. METHODS: Routine postoperative cardiac surgery patients were studied. Lactic acid levels were greater than 4 mmol/L in study patients and less than 4 mmol/L in controls. Polymerase chain reaction-based techniques were used to examine carriage of tumor necrosis factor beta (TNF beta), TNF G-308A, and interleukin 10 (IL-10) G-1082A alleles. RESULTS: Demographic characteristics and details of surgery were similar for 30 control and 21 study patients. Lactic acid levels after intensive care admission changed over time and were related to both TNF-beta and IL-10 G-1082A polymorphisms. All 4 study patients homozygous for TNF-beta1 and carrying an IL-10-1082A allele developed lactic acidosis (p = 0.02). There was no relation between the rate of epinephrine infusion or duration of cardiopulmonary bypass and lactic acid levels. CONCLUSIONS: Genetic factors have a role in the development of lactic acidosis after cardiac surgery. PMID- 12078790 TI - Retrograde autologous priming of the cardiopulmonary bypass circuit reduces blood transfusion after coronary artery surgery. AB - BACKGROUND: Hemodilution occurring with cardiopulmonary bypass imposes a risk for blood transfusion. Autologous priming of the cardiopulmonary bypass circuit at the initiation of bypass partially replaces the priming solution with autologous blood. We examined the efficacy of autologous priming of the circuit in reducing blood transfusion. METHODS: One hundred and four patients were entered into a prospective, randomized, controlled study. Initiation of cardiopulmonary bypass was with or without autologous priming. RESULTS: With autologous priming, a mean volume of 808.8 +/- 159.3 mL of priming solution was replaced with autologous blood. This allowed a higher hematocrit value on admission to the intensive care unit and at discharge from hospital. In all, 49% of the control group required a blood transfusion compared with 17% from the autologous priming group (p = 0.0007). The mean volume of blood transfused was 277.6 +/- 363.8 mL in the control group compared with 70.1 +/- 173.5 mL in the autologous priming group (p = 0.0005). CONCLUSIONS: Retrograde autologous priming of the bypass circuit reduces homologous blood transfusion owing to the reduction in bypass circuit priming volume. PMID- 12078791 TI - Mesenchymal stem cell implantation in a swine myocardial infarct model: engraftment and functional effects. AB - BACKGROUND: A novel therapeutic option for the treatment of acute myocardial infarction involves the use of mesenchymal stem cells (MSCs). The purpose of this study was to investigate whether implantation of autologous MSCs results in sustained engraftment, myogenic differentiation, and improved cardiac function in a swine myocardial infarct model. METHODS: MSCs were isolated and expanded from bone marrow aspirates of 14 domestic swine. A 60-minute left anterior descending artery occlusion was used to produce anterior wall infarction. Piezoelectric crystals were placed within the ischemic region for measurement of regional wall thickness and contractile function. Two weeks later animals autologous, Di-I labeled MSCs (6 x 10(7)) were implanted into the infarct by direct injection. Hemodynamic and functional measurements were obtained weekly until the time of sacrifice. Immunohistochemistry was used to assess MSC engraftment and myogenic differentiation. RESULTS: Microscopic analysis showed robust engraftment of MSCs in all treated animals. Expression of muscle-specific proteins was seen as early as 2 weeks and could be identified in all animals at sacrifice. The degree of contractile dysfunction was significantly attenuated at 4 weeks in animals implanted with MSCs (5.4% +/- 2.2% versus -3.37% +/- 2.7% in control). In addition, the extent of wall thinning after myocardial infarction was markedly reduced in treated animals. CONCLUSIONS: Mesenchymal stem cells are capable of engraftment in host myocardium, demonstrate expression of muscle specific proteins, and may attenuate contractile dysfunction and pathologic thinning in this model of left ventricular wall infarction. MSC cardiomyoplasty may have significant clinical potential in attenuating the pathology associated with myocardial infarction. PMID- 12078792 TI - Avoiding ischemia in latissimus dorsi muscle grafts: electrical prestimulation versus vascular delay. AB - BACKGROUND: Surgical mobilization of the latissimus dorsi muscle produces regional ischemic damage that may compromise its function in clinical applications such as cardiomyoplasty. We compared the effectiveness of two procedures designed to maintain blood flow throughout the mobilized muscle. METHODS: Adult pigs were assigned to two experimental groups: an electrically prestimulated group (n = 10) and a vascular delay group (n = 10). In the prestimulated group the left latissimus dorsi muscle was activated in situ at 2 Hz for 24 h/d. In the vascular delay group, the intercostal perforating arteries to the left latissimus dorsi muscle were divided. Two weeks later, hyperemic blood flow was measured by means of fluorescent microspheres immediately before and after mobilizing the latissimus dorsi muscle and again after recovery for a further 2 days. RESULTS: In the prestimulated group, blood flow was not significantly depressed in any region of the muscle immediately after mobilization, and blood flow increased significantly in proximal (p = 0.01), middle (p = 0.02), and distal (p = 0.007) regions following recovery. In muscles subjected to vascular delay the proximal and middle regions showed no significant changes in blood flow after mobilization or recovery, but flow in the distal region was 50% lower after mobilization (p = 0.003), and it remained significantly depressed even after recovery (p = 0.008). CONCLUSIONS: Prestimulation was significantly more effective than vascular delay in preserving distal blood flow. Because it is also less invasive and initiates metabolic transformation before mobilization, this technique should allow cardiac assistance to be introduced at an earlier postoperative stage without compromising the viability of the grafted muscle. PMID- 12078793 TI - Platelet activation, aggregation, and life span in calves implanted with axial flow ventricular assist devices. AB - BACKGROUND: A variety of rotary blood pumps are under development worldwide to serve as chronic ventricular assist devices (VADs). Historically VADs have been associated with thrombotic and thromboembolic complications, yet the ability to evaluate the thrombotic process in preclinical device testing has been limited. METHODS: We have developed and applied flow cytometric assays for activated platelets, platelet microaggregates, and platelet life span and consumption to calves implanted with an axial flow VAD and calves undergoing a sham surgical procedure. RESULTS: Surgical sham calves had significant increases in circulating activated platelets (p < 0.05) that resolved within 17 days, and no increases in circulating platelet microaggregates. Calves with uneventful VAD implant periods had early transient elevations in platelet microaggregates and prolonged elevations in activated platelets that did not recover to preoperative values during the study. Daily platelet consumption in VAD implanted calves was increased by 20% +/- 3%. Calves with thrombotic deposition within the VAD and elevated thromboembolism observed at autopsy experienced increases in circulating activated platelets and microaggregates at the end of the implant period when VAD flow decreased. CONCLUSIONS: This study demonstrates the ability of flow cytometry-based platelet assays to differentiate VAD implant operations from VAD support, and suggests differences that exist between uneventful VAD support and support with complications. These techniques should have value in evaluating other cardiovascular devices undergoing preclinical testing and provide insight into the temporal impact of these devices on the hemostatic system. PMID- 12078794 TI - Global cardiac-specific transgene expression using cardiopulmonary bypass with cardiac isolation. AB - BACKGROUND: The available techniques for intravascular gene delivery to the heart are inefficient and not organ-specific. Yet, effective treatment of heart failure will likely require transgene expression by the majority of cardiac myocytes. To address this problem, we developed a novel cannulation technique that achieves efficient isolation of the heart in situ using separate cardiopulmonary bypass (CPB) circuits for the heart and body in dogs. METHODS: The arterial inflow and venous effluent from the two circuits were physically isolated. The efficiency of separation was 98% to 99% in three preliminary experiments using Evans Blue dye labeled albumin. In 6 dogs, the cardiac circuit was perfused with oxygenated crystalloid cardioplegia at 37 degrees C containing approximately 4 x 10(11) particles of an adenovirus encoding LacZ (AdCMVLacZ) with a perfusion pressure of 170 to 200 mm Hg for 15 minutes allowing virus to recirculate through the heart approximately 15 times. Cross-clamp time was 26 +/- 2 minutes and CPB time was 90 +/- 3 minutes. RESULTS: Five animals survived and were euthanized at 7 days. Beta galactosidase activities measured using a chemiluminescent assay were three orders of magnitude higher in all areas of the heart than in the liver. Histological analyses revealed heterogeneous X-Gal staining of myocytes in all areas of the myocardium. CONCLUSIONS: Despite using a constitutive promoter, this technique yields relatively cardiac-specific transgene expression and is potentially translatable to clinical applications. Future studies will allow for further optimization of the conditions necessary for vector-mediated gene delivery to the heart. PMID- 12078795 TI - Graft-versus-host disease-type colitis: an unusual association of malignant thymoma. AB - We present the case of a patient with a malignant thymoma associated with a graft versus-host disease-type colitis, of which there has only been a solitary case report in the world literature. Complete surgical resection of the thymoma was achieved; however, the gastrointestinal symptoms persisted. PMID- 12078796 TI - Bronchial transection resulting from trivial blunt chest trauma. AB - A 3-year-old boy with a trivial blunt chest injury presented with a massive bronchopleural leak without any hilar vascular injury. On emergency exploration he had a complete transection of the right main bronchus. An end-to-end anastomosis of the transected bronchial ends was performed. At 1-month clinical and radiologic follow-up, the anastomosis had healed well. PMID- 12078797 TI - Seat belt-related chondrosternal disruption with lung herniation. AB - A case of blunt chest trauma resulting in anterior chondrosternal separation with right lung herniation and hemothorax is presented. The injury is related to the use of a seat belt restraint. The patient underwent surgical repair with polytetrafluoroethylene chest wall reconstruction. Postoperative recovery was complicated by respiratory insufficiency due to underlying pulmonary contusion and multiple rib fractures. PMID- 12078798 TI - Incidental finding of myeloproliferative disorders during sternotomy. AB - We report 2 cases of myeloproliferative disorders discovered incidentally at the time of routine coronary bypass surgery. Suspicion of abnormal bone marrow tissue upon performing sternotomy and subsequent sampling for frozen section made the diagnosis. The surgical plan was changed, and partial revascularization without cardiopulmonary bypass was performed. PMID- 12078799 TI - Giant mediastinal chordoma. AB - A chordoma is a slow-growing tumor representing about 5% of all malignant bone tumors. Mediastinal chordoma is very rare. We report a giant thoracic chordoma in a 32-year-old woman who presented with chest pain, progressive dyspnea, and cough. Open biopsy confirmed a definitive preoperative diagnosis, and complete surgical excision of the tumor was accomplished. PMID- 12078800 TI - Empyema necessitatis into the retroperitoneal space. AB - Empyema necessitatis is a rare complication of tuberculous empyema. We present a very rare case of empyema necessitatis into the retroperitoneal space through the diaphragm. The fistula between the thoracic empyema cavity and the retroperitoneal abscess was clearly identified by magnetic resonance imaging. PMID- 12078801 TI - Chylothorax associated with massive osteolysis (Gorham's syndrome). AB - We report a 15-year-old boy with bilateral chylothorax complicating Gorham's syndrome. Thoracic duct ligation failed to prevent fluid reaccumulation. The patient died of lymphocytopenia. Autopsy revealed vascular proliferation in the parietal and visceral pleura as well as in the ribs, which seemed to cause persistent chylothorax responsible for the poor prognosis of this patient. We reviewed treatments provided to 22 patients reported in the literature with Gorham's syndrome and chylothorax. PMID- 12078802 TI - Large cell neuroendocrine carcinoma: an unusual presentation. AB - Primary lung cancer presenting as a pulmonary artery mass is unusual. We describe such a presentation in a patient with a large cell neuroendocrine carcinoma of the lung, its evaluation, and its treatment. PMID- 12078803 TI - Primary mesenchymal chondrosarcoma of the lung. AB - Mesenchymal chondrosarcoma has been well documented in the somatic soft tissue and bone. It is a rare subtype of chondrosarcoma characterized by the presence of islands of chondroid or by less osteoid tissue enmeshed within dense sheets of primitive small blue mesenchymal cells with hemangiopericytoma-like vessels, or by both. The vast majority of previously published pulmonary mesenchymal chondrosarcoma was metastatic. To the best of our knowledge, only one case of primary pulmonary mesenchymal chondrosarcoma has been described in the literature. Herein, we report the second case of primary mesenchymal chondrosarcoma of the lung and emphasize that biopsy may yield only nonspecific small blue cells, whereas a detailed evaluation of the resected specimen allows definite diagnosis of this rare lung tumor. PMID- 12078804 TI - Pulmonary artery stenting for recurrent lung cancer after left pneumonectomy. AB - We present a case of a patient with stenosis of the pulmonary artery which was successfully treated by implantation of a vascular endoprosthesis. A 50-year-old man underwent left pneumonectomy for lung cancer. Eleven months later, a computed tomographic scan revealed a soft tissue mass in the mediastinum and there was severe stenosis of the remaining right main pulmonary artery. A self-expandable vascular endoprosthesis was implanted in the stenotic portion. We used percutaneous cardiopulmonary support (PCPS) during the procedure. We recommend the technique of pulmonary artery stenting using PCPS as efficacious and safe. PMID- 12078805 TI - Pulmonary infarct cytologically mimicking adenocarcinoma of the lung. AB - We describe a patient who underwent a right lower lobectomy for a lung lesion with a preoperative diagnosis of adenocarcinoma made on sputum cytology. Pathologic assessment of the resected specimen revealed a pulmonary infarct with no evidence of malignancy. All thoracic surgeons should be aware of this diagnostic pitfall. A positive sputum specimen should not always be regarded as definitive evidence of malignancy. Instead a full assessment of all available information should be made if unnecessary surgery is to be avoided. PMID- 12078806 TI - Glucocorticoids and hippocampal atrophy after heart transplantation. AB - The glucocorticoid cascade hypothesis proposes that hippocampal atrophy may result from excessive steroid exposure. Although demonstrated in animal models and some human hypercortisolemic states, hippocampal damage as a possible consequence of posttransplant steroid immunosuppression has not been investigated in human heart transplant recipients. We report a case of a 37-year-old female heart transplant recipient who had the clinical, neuropsychiatric, and neuroimaging findings consistent with hippocampal atrophy after 5 years of steroid exposure. PMID- 12078807 TI - Spontaneous disappearance of a false aneurysm after iatrogenic ruptured ventricle. AB - We report the case of a 64-year-old woman who had an atypical subendocardial aneurysm, a space between the internal patch, which was used for the repair of a left ventricular rupture after mitral valve replacement, and myocardium, which filled with blood during the diastole phase. During the follow-up period, the aneurysm spontaneously disappeared. This case endorses combining internal and external approaches to repair a left ventricular rupture after mitral valve replacement. PMID- 12078808 TI - Autograft aortic arch extension and sleeve resection for bronchial compression after interrupted aortic arch repair. AB - Successful correction of bronchial compression and severe bronchomalacia complicating repair of interrupted aortic arch was achieved using transverse aortic arch extension with a pulmonary artery autograft and left bronchial sleeve resection. This procedure increased space within the aortic arch and eliminated bronchial narrowing with excellent results. PMID- 12078809 TI - Anterior spinal artery syndrome after elective coronary artery bypass grafting. AB - A 65-year-old patient with ischemic heart disease and hypertensive nephropathy had paraplegia develop after elective coronary artery revascularization caused by anterior spinal infarction. Spinal complications are rare after coronary artery bypass grafting. The possible mechanisms of spinal cord injury are discussed, and the relevant literature is reviewed. PMID- 12078810 TI - The Ross procedure for endocarditis in a 4-month-old infant. AB - Streptococcal endocarditis in an infant is rare. We report a case of acute aortic valve endocarditis with abscess and aorta-to-right atrial fistula formation. This 4-month-old infant with a structurally normal heart had been previously well. The child was successfully treated with the Ross procedure and remains well 13 months postoperatively. PMID- 12078811 TI - Huge sinus of Valsalva aneurysm causing mitral valve incompetence. AB - We describe a case of a large sinus of Valsalva aneurysm originating from the noncoronary sinus. The aneurysm compressed the roof of the left atrium rendering the annulus and the anterior leaflet of the mitral valve severely distorted and, as a result, incompetent. The neck of the aneurysm was closed with a patch from the side of the aortic sinus, but we had to replace the mitral valve as the distorted structure did not resume its original shape even though we opened the aneurysm and debrided all thrombotic material inside the aneurysm. PMID- 12078812 TI - Pulmonary thromboendarterectomy in a patient with giant cell arteritis. AB - This report describes the case of a young woman presenting with signs and symptoms of chronic thromboembolic pulmonary hypertension who underwent pulmonary thromboendarterectomy (PTE) with concomitant coronary artery bypass. She died in the intensive care unit 1 day postoperatively. At autopsy the patient was found to have giant cell arteritis of the pulmonary arteries and ascending aorta. It is important to differentiate this disease from chronic thromboembolic pulmonary hypertension because its management and that of systemic vasculitis differs considerably. PMID- 12078813 TI - Stenosed kinking of the left main coronary artery combined with an aneurysm of the ascending aorta. AB - Stenosed kinking of the left main coronary artery is described in a 60-year-old man with an aneurysm of the ascending aorta and severe aortic valve insufficiency. An elective complete ascending aorta replacement using a valved composite graft was performed along with a shortening and direct implantation of the left main coronary artery. The right coronary artery was reimplanted as a Carrel-button. PMID- 12078814 TI - Aneurysm of the left atrial appendage. AB - We present the case of a 33-year-old woman with atrial tachyarrhythmias and chest pain. The transthoracic echocardiography demonstrated an intrapericardial liquid mass confirmed as an aneurysm of the left atrial appendage by a nuclear magnetic imaging study. Aneurysmectomy was performed with the assistance of cardiopulmonary bypass with a bilateral submammary skin incision and subsequent median sternotomy. The patient had an uneventful postoperative course. We suggest aneurysmectomy aided by cardiopulmonary bypass as a safer method of treatment for this rare cardiac anomaly. PMID- 12078815 TI - Postoperative transient internal thoracic artery pseudoaneurysm. PMID- 12078816 TI - A prominent Chiari network. PMID- 12078817 TI - Double valve replacement through an aorto-annulo-atriotomy using an aortic-valved graft in a mitral position. AB - We present a modified technique for the reconstruction of the intervalvular fibrous body in double-valve replacement through an aorto-annulo-atriotomy. This technique allows the surgeon to enlarge and reconstruct both annuli by using a tailored aortic-valved conduit in a mitral position. PMID- 12078818 TI - Double-switch Ross procedure. AB - Aortic root replacement with pulmonary autograft (Ross procedure) is a valuable technique. However, the best material for right ventricular outflow tract reconstruction remains controversial. We report on the experience with use of an aortic autograft with reimplantation of the diseased aortic valve for right ventricular outflow tract reconstruction in 3 patients with satisfactory result. PMID- 12078819 TI - A new pattern for using both thoracic arteries to revascularize the entire heart: the pi-graft. AB - We present a complex graft for total arterial revascularization based on bilateral skeletonized internal thoracic arteries (ITA). The lower two-thirds of the free right ITA is anastomosed to the proximal segment of the left in situ ITA using the T-graft technique (Tector-Barner-Calafiore). The free, transected distal part of the left ITA is then anastomosed end-to-side on free right ITA (T on-T anastomosis). In addition, the technique may use another graft extending the proximal third of the in situ right ITA with the free radial artery for right sided revascularization. The entire operation can be performed off-pump to avoid any procedure on the ascending aorta. PMID- 12078820 TI - Angiographic results after the use of a sutureless aortic connector for proximal vein graft anastomoses. AB - The sutureless aortic connector system was recently developed to create anastomoses between the aorta and saphenous vein grafts for coronary artery bypass grafting. Ten consecutive patients scheduled for coronary artery bypass grafting were studied; in each patient one proximal anastomosis was performed using the aortic connector and another was sutured in the conventional manner to serve as the patient's own control. One graft in the sutured group had a significant stenosis and one of the grafts in which the aortic connector was used, was occluded. In this limited study the use of the sutureless connector system for proximal anastomosis shows satisfactory early graft patency (90%). PMID- 12078821 TI - Tracheal replacement: a critical review. AB - This review discusses the need for tracheal replacement, distinct from resection with primary anastomosis, the requirements for replacement, and the many efforts over the past century to accomplish this goal experimentally and clinically. Approaches have included use of foreign materials, nonviable tissue, autogenous tissue, tissue engineering, and transplantation. Biological problems in each category are noted. PMID- 12078822 TI - Determining the extent of cognitive change after coronary surgery: a review of statistical procedures. AB - Currently, cognitive decline after coronary surgery is said to be significant if the individual's postoperative test score is at least 1 standard deviation (SD) worse than their preoperative score. This "1-SD" technique fails to account for factors that may confound interpretation of serially acquired cognitive test scores, including regression to the mean, measurement error caused by poor test retest reliability, and practice effects. We review the many alternative and potentially superior statistical techniques that have been described in the neuropsychologic and psychiatric literature for differentiating "true" changes in cognitive test score from changes caused by these factors. PMID- 12078823 TI - Photodynamic therapy in thoracic surgery. PMID- 12078824 TI - Thoracic surgery workforce: snapshot at the end of the twentieth century and implications for the new millennium. AB - BACKGROUND: An appropriately sized physician workforce is an essential component for healthcare planning. Contemporary economic forces and the impact of managed care have stimulated renewed interest in understanding and monitoring workforce issues. METHODS: Between August and December of 1999, the Workforce Committee of the American Association for Thoracic Surgery and The Society of Thoracic Surgeons conducted a survey detailing demographic, geographic, and socioeconomic practice characteristics of the membership. RESULTS: The 2,515 returned surveys represented a 62.6% return rate (+/- 1.6% margin of error), providing a highly statistically valid sample. Active thoracic surgeons are a mean of 50 years old, 98% men, with a high level of career satisfaction. Length of training and educational debt has been escalating. Adult cardiac surgical case volumes have increased (mean, 225 cases) and 52% of adult cardiac surgeons perform general thoracic procedures. Surgeons work 67 hours/week and 47 weeks/year. Most are organized in single specialty groups (50%) in urban areas (59%) as for-profit corporations (77%). Half of the respondents will be retired a mean of 13 years from now corresponding to a median calendar year of 2011. CONCLUSIONS: This survey highlights significant workforce retirement during the next 10 to 15 years. Currently, the workforce in thoracic surgery appears "right sized." These valuable data provide a profile of the specialty. The information enhances our ability to strategically plan. PMID- 12078826 TI - Reduction in tracheostomy-associated risk of mediastinitis by routine use of percutaneous tracheostomy. PMID- 12078825 TI - Acute amiodarone-induced pulmonary toxicity after lung operation. PMID- 12078827 TI - Blood hemoglobin as an independent prognostic factor in surgically resected stages I and II non-small cell lung cancer patients. PMID- 12078828 TI - Combined aortic and mitral valve operations with transection of the ascending aorta. PMID- 12078829 TI - Is off-pump bypass better? Adverse effects of on-pump conversion. PMID- 12078830 TI - Vegetectomy in Brucella endocarditis. PMID- 12078831 TI - Retrograde cerebral perfusion. PMID- 12078832 TI - Hormone replacement therapy and the prevention of cardiovascular disease. AB - Cardiovascular disease (CVD) is the primary killer of both men and women in Western societies. The implementation of preventive strategies has led to a fall in the rate of CVD, but there is still much to be achieved. Proven interventional strategies are largely under-utilized, and the search continues for further promising interventions. HRT appears to reduce CVD in post-menopausal women, based on observational data supported by plethora of evidence for the beneficial cardiovascular effects of estrogen. However, a recent controlled trial in post menopausal women with established CVD has shown that a specific combined oral HRT regimen did not reduce, and may even contribute to, an early increase in cardiovascular events, suggesting that HRT is inappropriate in secondary prevention. HRT may be useful in the primary prevention of CVD, yet observational data that suggested cardiovascular benefit with HRT also suggests that 80% of CVD in women could be eliminated by lifestyle modification, without the attendant risks of HRT including thrombosis and (potentially) breast cancer. At present, it is arguable that the evidence is inadequate to recommend HRT solely for the purpose of CVD prevention, and that the challenge for the health professional should be appropriate utilization of established preventative therapies, with further research into the potential role of HRT and estrogen-receptor modulators. PMID- 12078833 TI - Transmission of male infertility to future generations: lessons from the Y chromosome. AB - The introduction of ICSI and testicular sperm extraction (TESE) has allowed many infertile men to father children. The biggest concern about the wide use of these techniques is the health of the resulting offspring, in particular their fertility status. If the spermatogenic defect is genetic in origin, there is potential risk of transmitting this defect to future offspring. The most frequently documented genetic cause of male infertility is a Y chromosome deletion. The Y chromosome has acquired a large number of testis-specific genes during recent evolution, and deletions causing infertility take out a number of these genes. These deletions have been shown to be transmitted to 100% of male offspring. Also, absence of an aberration on the Y chromosome does not rule out a genetic cause of the infertility phenotype, as there are many other genes involved in spermatogenesis elsewhere in the genome, and current mapping techniques--especially on the Y chromosome--can miss many aberrations. More detailed studies of these spermatogenesis genes, which are now possible because of more precise sequence-based mapping, will lead to improved understanding of the genetic basis of male infertility and enable proper counselling of patients undergoing ICSI in the future. PMID- 12078834 TI - Long-term health consequences of PCOS. AB - The prevalence of cardiovascular risk factors, insulin resistance/diabetes and/or uterine pathology appears to be increased in women with polycystic ovarian syndrome (PCOS), although more outcome studies are necessary to determine incidence. Data pertaining to some of the potential long-term health consequences associated with PCOS are summarized. Medline, Current Contents and PubMed were searched for studies from the time of our original interest in this issue in 1980 to the present. The review is limited to published human data. The current literature indicate that women with this syndrome cluster risk factors for premature morbidity and mortality. Large multi-site co-operative studies are necessary to evaluate the long-term health outcomes. PMID- 12078835 TI - Follicle culture in reproductive toxicology: a tool for in-vitro testing of ovarian function? AB - Public opinion and official bodies place great emphasis on the reduction, refinement and replacement of the use of laboratory animals in testing protocols. In the field of toxicology, major efforts are being made to commit to this goal. The testing of reproductive function is currently still performed by in-vivo tests, mainly in rodents. In the past, follicle culture models were developed for the in-vitro production of mature oocytes and used to study the process of folliculogenesis and oogenesis in vitro. These culture systems might be able to acquire a place in fertility testing, replacing in-vivo studies for ovarian function and female gamete quality testing. The proficiency data from a well characterized follicle culture system suggest that this bioassay might be of potential use for in-vitro screening of xenobiotic substances affecting ovarian function and fertility. PMID- 12078836 TI - New insights into the mechanisms underlying hydrosalpinx fluid formation and its adverse effect on IVF outcome. AB - The adverse effects of hydrosalpinx on the outcome of IVF have been well documented; however, the causes for impaired implantation in patients with hydrosalpinx are poorly understood. Hydrosalpinx fluid has been shown to be toxic to mouse embryos but not human embryos, and this has become a topic of intense debate. An understanding of the mechanisms underlying hydrosalpinx formation following pelvic inflammatory disease appears to be essential in elucidating the causes for reduced implantation in hydrosalpinx patients and providing more rational treatments. This review discusses the mechanisms underlying hydrosalpinx formation and its adverse effect on IVF outcome, with new insights into possible involvement of Fallopian tube epithelial transporters and ion channels, particularly the cystic fibrosis transmembrane conductance regulator (CFTR). Possible links between Chlamydia trachomatis in pelvic inflammatory disease and the subsequent CFTR-mediated events in hydrosalpinx formation leading to infertility in hydrosalpinx are proposed. The causes of reduced implantation, particularly in patients with visible hydrosalpinges shown on ultrasound scanning, are re-examined in light of these possible mechanisms. PMID- 12078837 TI - An international survey of the health economics of IVF and ICSI. AB - The health economics of IVF and ICSI involve assessments of utilization, cost, cost-effectiveness and ability to pay. In 48 countries, utilization averaged 289 IVF/ICSI cycles per million of population per annum, ranging from two in Kazachstan, to 1657 in Israel. Higher national utilization of IVF/ICSI was associated with higher quality of health services, as indicated by lower infant mortality rates. IVF and ICSI are scientifically demanding and personnel intensive, and are therefore expensive procedures. The average cost per IVF/ICSI cycle in 2002 would be US$9547 in the USA, and US$3518 in 25 other countries. Price elasticity estimates suggest that a 10% decrease in IVF/ICSI cost would generate a 30% increase in utilization. The average cost-effectiveness ratios in 2002 would be US$58,394 per live birth in the USA, and US$22,048 in other countries. In three randomized controlled trials, incremental costs per additional live birth with IVF compared with conventional therapy were US$ 26,586, $79,472 and $47,749. The national costs of IVF/ICSI treatment would be US$1.00 per capita in one current model, but the costs to individual couples range from 10% of annual household expenditures in European countries to 25% in Canada and the USA. PMID- 12078838 TI - The use of GnRH antagonists in ovarian stimulation. AB - GnRH antagonists induce a rapid decrease in LH and FSH, preventing and interrupting LH surges. Their properties do not require a desensitization period, and this allows their use in the late follicular phase. GnRH antagonists could replace GnRH agonists in controlled ovarian stimulation without their side effects and their long desensitization period. Two protocols for assisted reproduction technology (ART) cycles were designed: the single-dose protocol allies simplicity and efficacy, while the multiple-dose protocol is efficient and could reduce monitoring of the cycle, though compliance is mandatory. A review of the available literature on GnRH antagonists in ART cycles is presented, focusing on phase III controlled trials and ART results. Both protocols using GnRH antagonists were associated with the need for a smaller dose of gonadotrophin, a shorter stimulation period and a lower incidence of ovarian hyperstimulation syndrome (OHSS), albeit with statistically comparable pregnancy rates. A trend is observed in all studies showing a lower pregnancy rates in GnRH antagonist cycles as compared with GnRH agonist cycles. The role of the lower number of embryos, and the potential adverse effects of GnRH antagonists on endometrium or follicle must be studied. More cycles using GnRH antagonists are necessary to confirm their equivalent pregnancy rates. There is room for improvement in both protocols with regard to scheduling, antagonist dose level and the timing of its administration. Until further studies have been conducted, luteal support seems to remain mandatory. Perinatal outcome appears similar to that with other stimulation regimens. Triggering of ovulation can be obtained with GnRH agonist for patients at risk of OHSS. With regard to GnRH antagonists, questions remain regarding pregnancy rates, the indications of their use in patients with polycystic ovary syndrome or poor responders, and in ovarian stimulation outside IVF. PMID- 12078839 TI - A qualitative systematic review of coasting, a procedure to avoid ovarian hyperstimulation syndrome in IVF patients. AB - 'Coasting', a method which consists of stopping exogenous gonadotrophins and postponing HCG administration until the patient's serum estradiol (E2) level decreases, is often used to prevent ovarian hyperstimulation syndrome (OHSS). We conducted a systematic review to analyse whether there is sufficient evidence to justify the general acceptance of coasting. The studies, which involved 493 patients in 12 studies, are very heterogeneous in the characteristics and number of patients in the ovulation stimulation schemes. The study designs, control groups, selection criteria for coasting and the OHSS classifications were variable. In most studies a threshold value of E2 was used (often 3000 pg/ml) and/or the number of follicles were considered. The fertilization rates (36.7 71%) and the pregnancy rates (20-57%) were acceptable in terms of IVF results in comparison with those of other large IVF databanks. In 16% of the cycles, ascites was described and 2.5% of the patients required hospitalization. In conclusion, while coasting does not avoid totally the risk of OHSS, it decreases its incidence in high-risk patients. Many questions remain unanswered about how coasting should be managed, and we suggest that a randomized prospective multicentre study is required. PMID- 12078840 TI - A case of elastosis perforans serpiginosa. AB - A 17-year-old white boy with no underlying connective tissue disorders presented with flat-topped annular plaques, with slight central atrophy on the bilateral neck. Results from histopathology revealed changes consistent with elastosis perforans serpiginosa (EPS). The idiopathic form of EPS occurs rarely in children. We report a patient with this rare pediatric diagnosis and review the literature. PMID- 12078842 TI - Familial ulcerative pyoderma gangrenosum: a report of 2 kindred. AB - Pyoderma gangrenosum is a rare, chronic ulcerative skin disease. It is a diagnosis of exclusion, after ruling out other causes of cutaneous ulceration. The etiology of pyoderma gangrenosum is poorly understood but is likely multifactorial. We describe 2 families affected by ulcerative pyoderma gangrenosum. This familial clustering suggests a possible genetic role in the development of pyoderma gangrenosum in some cases. PMID- 12078843 TI - What's eating you? Ixodes ticks. PMID- 12078841 TI - Photo quiz. Linear Scleroderma. PMID- 12078844 TI - Defining skin of color. AB - In the past few years, increasing attention has been given to skin disease in individuals with skin of color (also termed ethnic skin, racial skin, black skin, or pigmented skin). We will attempt to identify those individuals with skin of color. We use this article as an introduction to a recurring series that will address cutaneous disease affecting these individuals. PMID- 12078845 TI - Ant sting sporotrichosis. AB - Cutaneous sporotrichosis is an uncommon infection, usually reported as sporadic cases resulting from inoculation with sharp environmental vegetative matter. We report such a case of multiple primary inoculations acquired from Solenopsis (fire ant) stings in a 54-year-old white man. The patient was treated effectively with itraconazole 200 mg twice a day for 4 months. PMID- 12078846 TI - Lyme disease--part II: clinical features and treatment. AB - Lyme disease (LD) is the most common vector-borne disease in the United States. Generally presenting with a characteristic rash, myalgia, and fatigue, LD can progress to chronic arthritis, central nervous system manifestations, and cardiac abnormalities, if left untreated. The number of cases continues to rise each year. Early diagnosis and proper therapy are required to halt disease progression to late chronic stages. By adhering to simple guidelines, many potential cases of LD can be prevented. In this article, the second in a 2-part series on LD, we discuss clinical features and treatment. PMID- 12078847 TI - Grover disease (transient acantholytic dermatosis) and piebaldism. AB - A 35-year-old white man with lifelong stable white lesions on the anterior trunk and extremities presented with a pruritic papular eruption limited to the white patches. Results of a histologic examination led to a diagnosis of Grover disease (transient acantholytic dermatosis). To our knowledge, this article is the first to report an association between Grover disease and piebaldism or other depigmented disorders. We review the literature and speculate on the association between these conditions. PMID- 12078848 TI - Spiradenocarcinoma of the scalp. AB - Spiradenocarcinoma is an exceedingly rare malignant neoplasm with ductal differentiation. Many consider it to be an eccrine neoplasm, and others favor apocrine differentiation. In nearly all cases, spiradenocarcinoma is associated with a benign spiradenoma, with most lesions appearing on the trunk and extremities. We describe a patient who presented with a long-standing and previously asymptomatic scalp nodule that became tender and enlarged. After diagnosis of spiradenocarcinoma, the patient was referred for lymphoscintigraphy, sentinal lymph node biopsy, and Mohs micrographic surgery. PMID- 12078850 TI - Chronic acquired dyskeratotic papulosis of the face. AB - A Vietnam veteran who had been exposed to Agent Orange presented with a 28-year history of a papulonodular disease mainly affecting the face. Over those years, pin-sized papules had developed into dark nodules. Multiple biopsies of both fresh and mature papulonodules had been performed for routine histopathology and electron microscopy. Results had been positive for spongiosis with exocytosis, acanthosis, dyskeratotic keratinocytes, and marked incontinence of pigment, which together are suggestive of a previously unreported clinical entity that we term chronic acquired dyskeratotic papulosis. PMID- 12078849 TI - Patient and physician perspectives vary on atopic dermatitis. AB - We compared the written responses of physicians (n=303) and patients (n=961) from a nationwide US survey concerning atopic dermatitis (AD). Physicians, primarily dermatologists, responded to 32 questions, and patients responded to 44 questions about AD and its management. Most physicians and patients were in agreement regarding disease severity, the relative lack of effectiveness of over-the counter products, concerns about drug adverse effects, and the need for more patient support groups. However, physicians were more concerned about long-term adverse effects than were patients. Additionally, 91% of physicians versus 46% of patients rated prescription medications as "moderately" or "very" effective. Patients were generally pleased with their overall AD-related medical care: 42% were "a lot" or "very" satisfied, while only 8% were dissatisfied. Although physician and patient perceptions sometimes differed, both groups preferred treatments offering greater effectiveness, fewer adverse effects, and greater applicability to the pediatric population. PMID- 12078851 TI - Effect of topical benzoyl peroxide/clindamycin versus topical clindamycin and vehicle in the reduction of Propionibacterium acnes. AB - Propionibacterium acnes is one of the primary factors involved in the pathogenesis of acne vulgaris; proliferation of this bacteria is present in all patients with inflammatory lesions. Combination topical therapy with agents that have different but complementary antimicrobial mechanisms of action has the potential to increase efficacy and to prevent the emergence of resistant organisms. The onset of action and effectiveness of 3 topical preparations (benzoyl peroxide 5%/clindamycin phosphate 1% gel, clindamycin phosphate 1% solution, and vehicle gel) in reducing P acnes were compared in a randomized, open-label, evaluator-blinded, comparative trial involving 60 healthy volunteers who were free of acne but had high levels of facial P acnes. Treatment with benzoyl peroxide 5%/clindamycin phosphate 1% gel significantly (P<.001) reduced P acnes levels by >1 log10/cm2 from baseline (91% inhibition) 24 hours after application. Progressive declines were observed throughout the 2-week study period, with a 3 log10/cm2 reduction (99.9% inhibition) from baseline in P acnes at the end of the 2-week treatment period. In contrast, significant (P<.05) reductions from baseline in P acnes levels following treatment with clindamycin phosphate 1% solution were only observed at the last assessment period (2 weeks), with an average reduction of 0.64 log10/cm2 (77% inhibition). Patients receiving vehicle gel had no measurable reductions in P acnes from baseline. These results demonstrate that topical benzoyl peroxide 5%/clindamycin phosphate 1% gel produces rapid and clinically relevant reductions in P acnes greater than those produced by single-agent therapy. This activity is likely responsible for the quick onset of clinical efficacy produced by this combination regimen. PMID- 12078853 TI - Cross-priming of cytotoxic T cells promoted by apoptosis-inducing tumor cell reactive antibodies? AB - Humanizing xenogenic monoclonal antibodies (MAbs) by genetic engineering has greatly improved their therapeutic utility and efficacy. The chimeric CD20 MAb C2B8 (Rituximab) is a prominent representative of this new generation of therapeutic MAbs and has been proposed as a treatment of choice for recurrent follicular non-Hodgkin's lymphomas. Treatment of CD20+ B cells with MAb C2B8 triggers several cell-damaging actions including complement-mediated lysis (CDL), antibody-dependent cellular cytotoxicity (ADCC), and MAb-induced induction of apoptosis. We provide an overview of the most prominent mechanisms underlying the efficacy of antibody treatment. We introduce our concept of cross-priming of cytotoxic T-cell responses promoted by apoptosis incucing antibodies. Treatment of tumor cells with antibodies that are capable of inducing a proapoptotic signal via their cell surface target structure may not only contribute to their direct killing but also may induce cellular responses against the tumor, which may have a long-lasting protective effect. We report, using the example of C2B8 anti-CD20 treatment of lymphoma cells, that MAb C2B8-induced apoptosis of lymphoma cells not only kills these cells but also promotes uptake and cross-presentation of lymphoma cell-derived peptides by antigen-presenting dendritic cells (DC), induces maturation of DC, and allows the generation of specific CTL. PMID- 12078854 TI - Studies of ex vivo activated and expanded CD8+ NK-T cells in humans and mice. AB - Adoptive cellular therapy holds promise for improving the outcome of hematopoietic cell transplantation (HCT). At present, donor lymphocyte infusion post-HCT is efficacious for only a limited number of diseases, yet can induce significant graft versus host disease (GVHD). To improve the outcome of this approach, it would be beneficial to identify populations of T cells that retain graft versus tumor (GVT) effects with reduced propensity for GVHD. Here we describe studies of both human and murine expanded CIK cells or CD8+ NK-T cells. These related populations of cells are ex vivo activated and expanded T cells that express both T and NK markers. They can be generated from patients with malignancies and mediate cytotoxicity against autologous hematopoietic malignancies. Recent work in murine models show that these cells mediate cytotoxicity by using a perforin-granzyme and not through Fas ligand. In allogeneic stem cell transplantation experiments, large numbers of expanded CD8+ NK-T cells could be transplanted across major histocompatibility barriers without causing severe GVHD and GVT effects were retained. We conclude that expanded CD8+ NK-T cells are a promising form of cellular therapy in the allogeneic setting. PMID- 12078852 TI - Decryption of the retinoid death code in leukemia. AB - The recent elucidation of several molecular paradigms by which retinoids regulate growth, differentiation, and apoptosis highlights their promise as drugs for cancer therapy and prevention. Several novel signaling pathways by which retinoids induce cell death have been identified recently. They comprise (a) the induction by RARalpha-selective retinoids of the tumor-selective death ligand TRAIL that kills acute promyelocytic leukemia (APL) cells in a paracrine mode of action, which is the cause of retinoic acid-induced apoptosis after maturation: (b) a novel RARalpha-independent rexinoid-PKA cross-talk that induces maturation of both ATRA-sensitive and ATRA-resistant APL cells and does not invoke ligand induced alteration of PML-RARalpha signaling, stability, or compartmentalization; and (c) a novel rexinoid signaling pathway that triggers apoptosis of immature APL cells and may correspond to a default death pathway that is operative in the absence of "survival" factors. This rexinoid apoptosis is inhibited by RXR but not RAR antagonists and is distinct from that triggered by RAR agonists, which control cell maturation and postmaturation apoptosis. Here we discuss the promise of retinoids for cancer treatment and prevention with an emphasis on the recently identified mechanisms by which they control (cancer) cell proliferation. PMID- 12078856 TI - Induction of MCP-1 expression in airway epithelial cells: role of CCR2 receptor in airway epithelial injury. AB - The repair of an injured bronchial epithelial cell (BEC) monolayer requires proliferation and migration of BECs into the injured area. We hypothesized that BEC monolayer injury results in monocyte chemoattractant protein-1 (MCP-1) production, which initiates the repair process. BECs (BEAS-2B from ATCC) were utilized in this study. MCP-1 interacts with CCR2B receptor (CCR2B), resulting in cell proliferation, haptotaxis, and healing of the monolayer. Reverse transcriptase-polymerase chain reaction (RT-PCR) was employed to verify the presence of CCR2B. CCR2B was not merely present but also inducible by interleukin 2 (IL-2) and lipopolysaccharide (LPS). We demonstrated by immunohistochemistry that BECs express MCP-1 after injury and that receptor expression can be regulated by exposure to IL-2 and LPS. Haptotactic migration of cells was enhanced in the presence of MCP-1 and reduced in the presence of CCR2B antibody. This enhanced or depressed ability of the BECs to perform haptotactic migration was shown to be statistically significant (P < 0.05) when compared to controls. Finally, BECs proliferate in response to MCP-1 as proven by electric cell substrate impedance sensing (ECIS) technology. MCP-1-specific antibodies were shown to neutralize the MCP-1-mediated BEC proliferation. This cascade of events following injury to the bronchial epithelium may provide insight into the mechanism of the repair process. PMID- 12078855 TI - Study of autoimmunity in Klinefelter's syndrome and idiopathic hypogonadotropic hypogonadism. AB - Sex hormones play an important role in determining the progression and severity of autoimmune diseases, but the in vivo mechanisms underlying this relation are poorly understood. The main objective of current study has been to compare the changes in neuroendocrine immune features and autoantibody profile in male patients with hypogonadotropic and hypergonadotropic hypogonadism, and to determine the relationships between sex hormones and immunologic parameters. Thirty-seven male patients with Klinefelter's syndrome and 35 men with idiopathic hypogonadotropic hypogonadism who had no history of previous hormonal therapy and 30 healthy men were recruited in the study. Serum autoantibody profile, sex hormones, and immunologic parameters were studied. In conclusion, our findings suggest that both humoral and cellular immunity is enhanced in male hypogonadism. Klinefelter's syndrome patients also had increased frequency of antiextractable nuclear antibodies and anticardiolipin antibodies positivity compared to idiopathic hypogonadotropic hypogonadism patients. It is possible that testosterone deficiency and increased levels of estradiol are primary responsible factors for this enhanced autoantibody production in Klinefelter's syndrome patients. PMID- 12078857 TI - Interaction of disease-related antigen-reactive T-cell lines from multiple sclerosis patients with type IV collagen: role of integrin VLA-1 and effects of irradiation. AB - Multiple sclerosis (MS), a chronic demyelinating disease, is thought to be initiated by pathogenic T cells that transmigrate the vascular endothelium and enter the brain through vascular and parenchymal basement membranes (BM). Vaccination with T-cell lines reactive with myelin basic protein (MBP) and myelin oligodendrocytic glycoprotein (MOG) epitopes, expanded with interleukin-2 (IL-2), and attenuated by ionizing radiation is currently being evaluated as a therapeutic modality for this disease. We examined mechanisms potentially involved in pathogenic cell migration into the central nervous system (CNS) and the influence of irradiation on these processes. Seven of 7 autoantigen responsive T-cell lines from MS patients adhered to collagen IV, the major collagenous constituent of BMs. This adhesion was inhibited almost completely by monoclonal antibody (MAb) to very late antigen (VLA)-1 and partially by anti-VLA 2. T-cell lines from healthy donors adhered more variably to collagen IV. Furthermore, patient derived T cells actively transmigrated through a collagen IV gel toward medium containing TNF-a, in a process that was inhibited by MAbs to VLA-1. Ionizing radiation at the dose used in vaccine preparation, inhibited morphological polarization associated with migratory capability, induced integrin clustering on the cell membrane, and abrogated adhesion to collagen IV. These findings may have important implications for understanding the pathogenesis of MS and how irradiation of potentially pathogenic T cells produces a reagent with possible therapeutic effects in T-cell vaccination (TCV). PMID- 12078859 TI - Selective depletion of CD56(dim) NK cell subsets and maintenance of CD56(bright) NK cells in treatment-naive HIV-1-seropositive individuals. AB - Human immunodeficiency virus-I (HIV-1) infected patients show a gradual loss of natural killer (NK) cells that correlates with disease progression. However, the effect of HIV-1 infection on different NK cell subsets has not been fully characterized. In healthy individuals most NK cells are CD3-CD56+ and two different subpopulations, CD56(dim) and CD56(bright), can be distinguished by the mean fluorescence intensity. Although it was originally suggested that CD56(bright) NK cells represent the precursors of the CD56dim subpopulation, recent cumulative data indicate that CD56(bright) and CD56(dim) NK cells are phenotypically, functionally, and developmentally different NK cell subsets. In this study, the analysis of CD56(bright) and CD56(dim) NK subsets showed that neither the number nor the phenotype of CD56(bright) NK cells were significantly altered in treatment-naive HIV-1-infected individuals, whereas the number of CD56(dim) NK cells was decreased. We also have studied NK cell subsets defined by the expression of CD56 in combination with CD16, CD161, or CD94 molecules. Our results demonstrated a preferential decrease of CD3-CD56+ NK cells coexpressing CD16 and CD161 but lacking CD94 molecules. On the contrary an increased percentage of NK cells that do not express CD56 molecules but express CD16, CD161, or CD94 was also found in HIV-1-infected individuals. As it has been proposed that these CD56-negative NK cells expressing other NK cell receptors represent immature NK cells with low cytolytic capacity, our results support that a defective differentiation from immature CD56 negative NK cells to mature CD56(dim) NK cells occurs in HIV-1 infection. PMID- 12078860 TI - Case-control versus cohort studies in the investigation of childhood cancer. PMID- 12078858 TI - Cationic liposome-mediated enhanced generation of human HLA-restricted RSV specific CD8+ CTL+. AB - Generation of human CD8+ cytotoxic T-lymphocyte (CTL) activity against respiratory syncytial virus (RSV) using peripheral blood leukocytes (PBL) in vitro is inefficient. Lipofectamine, a polycationic liposome, previously shown to enhance the transfection efficiency of DNA in cells, was evaluated for enhancing RSV CTL activity. Stimulator cells were prepared by infecting human PBL with RSV with or without Lipofectamine for 3 hr and then transferred to responder cells. After 8 days of incubation, CTL lysis of autologous target cells infected with RSV (also treated with Lipofectamine) was determined in a 4-hr 5'chromium release assay. Lipofectamine treatment significantly enhanced HLA-restricted RSV-specific CD8+ CTL activity (up to sevenfold, P < 0.05-0.001). Lipofectamine treatment also enhanced cell surface RSV antigen expression and increased the frequencies of HLA A,B,C+/RSV+ and HLA-DR+/RSV+ leukocytes as demonstrated by flow cytometry. These results demonstrate the usefulness of cationic liposomes in augmenting cell surface antigen expression and increasing the efficiency of generation of human RSV-specific CD8+ CTL activity. PMID- 12078861 TI - Radiotherapeutic management of intracranial ependymoma. AB - Ependymomas comprise about 10% of all pediatric brain tumors. The most consistent prognostic factor for cure has been the extent of surgical resection. Radiation therapy is considered the standard adjuvant treatment, although there has been no randomized trial comparing surgery alone to surgery and postoperative radiotherapy. Craniospinal irradiation has been used in the past to treat these tumors; however, current data indicate that the most common pattern of failure is an isolated local relapse. Furthermore, prophylactic spinal irradiation has not been shown to prevent spinal dissemination. For this reason, most radiation oncologists currently employ localized radiotherapy fields. Available data indicate that doses greater than 45-50 Gy are needed and associated with better local control. Preliminary data using hyperfractionated radiotherapy doses of greater than 65 Gy indicate an improvement in progression-free survival for subtotally respected ependymoma. Chemotherapy can be used to delay institution of radiotherapy in children less than 3 years of age. The role of hemotherapy in older children needs to be further defined. PMID- 12078862 TI - Cutaneous melanoma in childhood and adolescence. AB - Using population-based data from the Surveillance Epidemiology and End Results Program of the National Cancer Institute, melanoma occurring during childhood was evaluated. Compared to adult cases of melanoma, childhood cases had a higher proportion of females (61%) and non-Caucasians (6.5%). The incidence of melanoma increased 85% among 15- to 19-year-olds from 1973 to 1996. Incidence for 15- to 19-year-olds was higher in southern (23.9/million) than northern registries (14.5/million). Non-Caucasians had 3-30% of the cases expected compared to Caucasians. Overall survival of children/adolescents with melanoma was 89% and 79% at 5 and 20 years postdiagnosis, respectively. The majority of deaths were directly attributed to melanoma (72%). PMID- 12078863 TI - Dextromethorphan is effective in the treatment of subacute methotrexate neurotoxicity. AB - Methotrexate-induced neurotoxicity (MTX-Ntox) is a frequent complication of methotrexate (MTX) therapy for patients with both malignant and inflammatory diseases. MTX-Ntox can occur after intrathecal MTX or after low-, intermediate-, or high-dose systemic administration. Symptoms can present in the acute, subacute, or late setting form, and can range from affective disorders, malaise, and headaches, to somnolence, focal neurologic deficits, and seizures. While the pathogenesis of MTX-Ntox is likely multifactorial, one potential biochemical pathway leading from MTX to neurotoxicity involves the folate dependent remethylation of homocysteine (Hcy). MTX therapy is known to cause elevations of both plasma and CSF Hcy. Hcy is directly toxic to vascular endothelium and it and its metabolites are excitatory agonists of the N-methyl-D-aspartate (NMDA) receptor. Competitive or noncompetitive antagonists might afford protection from or reversal of MTX-Ntox. Using high-performance liquid chromatography (HPLC) with coulometric electrochemical detection, the authors measured CSF Hcy in sequential patients with severe subacute MTX-Ntox. CSF Hcy was higher in these patients (n = 9, median = 0.93 microM) than in asymptomatic patients (n = 11, median 0.2 microM, p < .01). Five patients with severe subacute MTX-Ntox (most with dysarthria and/or hemiplegia) were treated with 1-2 mg/kg oral dextromethorphan (DM), a noncompetitive antagonist of the N-methyl-1-aspartate (NMDA) receptor. All five had resolution of symptoms. These data provide additional clinical support for elevated CSF Hcy in the induction of MTX-Ntox through activation of the NMDA-receptor. These data provide support for a placebo-controlled clinical trial to examine the ability of DM to prevent or alleviate MTX-Ntox. PMID- 12078864 TI - Pulsed high-dose dexamethasone therapy in children with chronic idiopathic thrombocytopenic purpura. AB - The effectiveness of pulsed high-dose oral dexamethasone therapy in children with refractory chronic idiopathic thrombocytopenic purpura (ITP) is evaluated. Thirteen children with severe chronic ITP were enrolled in the study from an outpatient pediatric hematology clinic (ages 2-14 years), 5 boys and 7 girls. They did not maintain a response to other forms of therapy (IVIg, Anti-D, conventional steroids, danazol) and one girl relapsed after splenectomy. Dexamethasone was administered orally at a dosage of 40 mg/M2/day (maximum 40 mg/day) for 4 consecutive days. The cycle was repeated once a month for 6 months. The immediate response to therapy was excellent as the mean platelet count at day 1 was 15 x 10(9)/L, while mean platelet count at day 4 was 158 x 10(9)/L. At the end of 6 cycles 3 patients maintained a platelet count of >150 x 10(9)/L and 4 patients showed partial response. At the end of the first year and second year (12 and 24 months after onset of treatment) 3 patients still had complete response, 3 patients had partial response, and 7 patients were failures. Six of the failures underwent splenectomy and one was shifted to dapsone, had no response, and refused splenectomy. Side effects were tolerable. They included bloating, nausea, vomiting, insomnia, anxiety, and depression, and transient glucosuria; however, they were not severe enough to discontinue the cycles. Mean duration of illness prior to start of dexamethasone was not significantly different in between responders and nonresponders. Dexamethasone given orally in high doses is an effective drug in achieving short-term platelet responses. Long term remission is obtained in nearly half the patients with well-established chronic ITP. Its effectiveness in almost half the patients, minimal side effects, and low cost indicate that this treatment should be considered in patients with chronic ITP who do not tolerate the disease well before considering splenectomy. PMID- 12078865 TI - Glutathione S-transferase and P-glycoprotein expressions in neuroblastoma. AB - This study was planned to evaluate the prognostic role of glutathione S transferase pi (GST-pi) and P-glycoprotein (P-gp) expressions in children with neuroblastoma. Sections from formalin-fixed paraffin-embedded tumor blocks from 52 neuroblastoma cases (17 with localized, 35 with advanced disease) were subjected to immunohistochemistry for P-gp and GST-pi expressions. The overall number of tumors positive for P-gp and GST-pi were 19 (36.5%) and 21 (40.4%), respectively. Twenty-two tumors were negative for both GST-pi and P-gp expressions, whereas 10 expressed both proteins. The distribution of staining status of samples in the groups of both proteins showed no significant difference. No relation between the expressions of both proteins and the clinical characteristics of the patients was demonstrable. The differenes between the survival rates of patients with positive and negative staining for P-gp expression were not statistically significant. Although 2 common mechanisms of multiple drug resistance, P-gp and GST-pi, might be responsible for drug resistance in neuroblastoma, this complex mechanism has no direct significant impact on prognosis. Multiple mechanisms at cellular levels are responsible for the resistance against antineoplastic therapies in neuroblastoma. PMID- 12078866 TI - Intracranial manifestation of osteosarcoma. AB - Involvement of the central nervous system in osteosarcoma is uncommon. These neoplasms are most often located at the metaphyses of tubular bones and rarely in flat bones of vertebra, ribs, pelvis, facial bones, or skull. Tumors of the latter bones may obviously spread into the cerebrum. Osteosarcomas primarily metastasize hematogenously to the lungs. Bone, lymph node, or brain metastases are mostly seen following or concomitantly with pulmonary metastatic disease. However, there are single cases of primary osteosarcoma of the brain parenchyma without bone association or tumor manifestation at other locations. Three illustrative cases highlight the diversity of the clinical presentation of cerebral osteosarcoma: a 22-year-old man with multiple brain metastases following late pulmonary relapse of an osteosarcoma of the tibia, a 31-year-old woman with an osteosarcoma of the left anterior cranial fossa arising from the skull base, and a 78-year-old man presenting with primary osteogenic sarcoma of the left frontal cerebral hemisphere. According to the current literature, 10-15% of all osteosarcoma patients experiencing relapse may beat risk for central nervous system metastases. To the authors' best knowledge, there are 11 cases of primary intracerebral or meningeal osteogenic sarcoma, including this case report, without any skeletal attachment. PMID- 12078867 TI - Differential chemosensitivity in a child with congenital relapsing acute lymphoblastic leukemia. AB - Described is a case of a boy with congenital acute lymphoblastic leukemia (ALL) with pre-pre-B-ALL immunophenotype, presenting as diarrhea, organomegaly, hyperleucocytosis of 1434 G/L, and tumor lysis syndrome. The lymphoblasts showed low proliferative activity and high in vitro drug sensitivity measured by the MTT assay. An excellent response to therapy was observed, but relapse ocurred 3 months later. On relapse, blasts showed extremely high drug resistance, high expression of P-glycoprotein, and high proliferative activity. The response to therapy was again positive, but a second relapse occurred in 1 month. The MTT assay indicated increasing drug resistance to all drugs. Cytogenetic analysis revealed deletion in 11q23 locus. This unfavorable case shows complex biology and differential drug resistance in congenital leukemia. PMID- 12078868 TI - A rare case of adenoviral fulminant hepatic necrosis after chemotherapy. AB - The authors report a rare case of fulminant adenoviral hepatic necrosis occurring after chemotherapy in a patient with a second relapse of acute myeloid leukemia. The literature is reviewed and the role of rapid viral diagnosis in the clinical management of this complication is discussed. A 10-year-old girl with relapsed acute myeloid leukemia after allogeneic bone marrow transplant underwent re induction chemotherapy with high-dose cytosine arabinoside and amsacrine. During induction she developed diarrhea and a marked coagulopathy, followed by fulminant hepatic failure and acute pre-renal failure. She rapidly deteriorated and died. A limited autopsy was performed. Adenovirus type 5 was cultured from ante mortem clinical samples and detected by polymerase chain reaction in postmortem samples of heart blood, lung, trachea, spleen, and liver. At autopsy, the liver demonstrated massive hepatic necrosis with positive immunofluorescence for adenovirus. Electron microscopy demonstrated intranuclear inclusions, typical of adenovirus. There was no evidence of pneumonia. Adenovirus can cause fulminant hepatic necrosis following chemotherapy in a nontransplant setting. If adenoviral disease is suspected, appropriate rapid viral studies should be undertaken, because early intervention with ribavirin or cidofovir may prevent rapid fulminant progression. Further studies on the role of antiviral therapy in this setting are warranted. PMID- 12078869 TI - Treatment of Kasabach-Merritt syndrome by megadose methylprednisolone. PMID- 12078870 TI - Piriformis muscle syndrome in 19 patients treated by tenotomy--a 1- to 16-year follow-up study. AB - We treated 19 patients with piriformis muscle syndrome. All patients complained of pain in the buttocks and the posterior thigh. Clinically the buttock was tender and passive stretching increased the pain. In ten patients skin sensation was reduced and three had a limp. All patients were treated with tenotomy. At follow-up after an average of 8 (1-16) years, eight patients had pain relief. Two thirds of the patients evaluated their clinical state as being better. PMID- 12078871 TI - The importance of spinopelvic parameters in patients with lumbar disc lesions. AB - Using a special software program we measured spinopelvic parameters on digitised radiographs of the entire spine and the pelvis of 50 patients with lumbar disc herniation and compared these with the same measurements on 30 healthy subjects. In the disc hernia group the patients had a relatively straight spine in the sagittal plane. The sacrum was more vertical, and the value of the lumbar lordosis was lower, as was the amplitude of the spinal curvatures, when compared with those of the healthy group. This results in a higher gravitational compressive force which may, in turn, lead to progressive degeneration of the discs. The anterior shift of the line of gravity may cause spinopelvic instability, and contraction of the posterior spinal muscles in trying to balance this disturbed spatial relationship may produce back pain. PMID- 12078872 TI - Beta-tricalcium phosphate ceramic as a bone substitute in orthopaedic surgery. AB - Synthetic beta-tricalcium phosphate ceramic (Biosorb, S.B.M., Lourdes, France) was used as a bone substitute in 110 patients whose mean age was 48 (14-83) years. There was a wide spectrum of indications, and the material was used alone in 86, combined with a corticocancellous bone autograft in 22, and combined with a corticocancellous allograft in two. The patients were assessed clinically and radiographically. Incorporation of the ceramic was excellent in 30, good in 51, and fair in 26. There were no poor results. In three patients the implanted material was removed because of infection. Incorporation was good or excellent in about 75% of all 110 patients. We consider beta-tricalcium phosphate ceramic to be the substitute of choice for medium-sized bone defects. PMID- 12078873 TI - Highly viscous sodium hyaluronate and joint lubrication. AB - We studied the natural lubrication mechanism of synovial joints. We determined the effect of sodium hyaluronate (HA) on lubricating joints without the normal lubrication mechanism. The coefficient of friction (CF) of fresh pig hip joints was measured with the cartilage intact, washed, scoured with gauze and finally with sandpaper, to model cartilage degradation. Three formulas of HA (8 x 10(5) daltons 1%, 20 x 10(5) daltons 1%, 20 x 10(5) daltons 1.5%) and physiologic saline were used as lubricants. We observed the cartilage using light microscopy (LM) and scanning electron microscopy (SEM). The latter showed that the most superficial layer observed in the washed joint was disrupted after gauze scouring. Compared with intact cartilage the CF did not increase with washing. CF increased more after scouring with sandpaper than with gauze. Each formula of HA decreased the CF of joints scoured with gauze, but only the two more viscous HA formulas decreased the CF of sandpaper-scoured joints. A negative correlation was found between the CF of the sandpaper-scoured joints and the logHA viscosity (r = -0.733, P = 0.0001), suggesting that HA with higher viscosity was more effective in lubricating the joints. PMID- 12078874 TI - Complications in the first 1,043 operations where self-reinforced poly-L-lactide implants were used solely for tissue fixation in orthopaedics and traumatology. AB - This is an analysis from one hospital of the first 1043 operations where pure self-reinforced poly-L-lactide (SR-PLLA) implants have been used alone. The operations were performed between 1988 and 1999 and included 407 orthopaedic patients and 636 trauma patients. There was a total of 107 complications. There were 21 infections but no sinus formation. Failure of fixation was seen in 46 patients. In 936 operations the healing was uneventful. PMID- 12078875 TI - Osteosarcoma occurring in osteogenesis imperfecta tarda. AB - The combination of osteogenesis imperfecta (OI) and osteosarcoma is rare, as reflected by the fact that there are only eight proven cases in the published English literature. We present here another case, one that was treated by modern oncological techniques and major limb salvage surgery in the form of distal femur endoprosthesis initially and later a total femur replacement. The patient is the only one of the series to have had major limb salvage and is the only--and longest--surviving patient (more than 6 years), thus disproving the probability that this combination of disorders is particularly lethal. We present the case history, review the literature, and look at the cases presented in the past. We contend that adequate awareness, early diagnosis, and prompt oncological management results in better prognosis for osteosarcoma occurring in OI. PMID- 12078876 TI - Patellar tendon avulsion after reconstruction of the anterior cruciate ligament. AB - We report a case of rupture of a reconstructed anterior cruciate ligament (ACL) combined with avulsion of the inferior patellar ligament. The two lesions were repaired during one operation using the quadriceps tendon as a graft for the damaged ACL. PMID- 12078877 TI - An assessment of cost-effectiveness studies in joint replacement. PMID- 12078878 TI - Total hip replacement in patients with Parkinson's disease. AB - From 1970 to 1994, 107 total hip arthroplasties (THAs) were performed in 98 patients with Parkinson's disease. The average age of the patients was 72 years. Preoperative diagnoses were osteoarthrosis in 58 hips, failed endoprosthesis in 19, aseptic loosening in ten, femoral neck fracture in 18, and other diagnoses in two. Milder neurological stages I-III were assigned to 96 patients, and tendon release for contracture was performed in eight patients. Of the 38 complications eight were urinary tract infections and six dislocations. Of these 15 occurred in the 58 primary THAs and 23 in the 49 nonprimary THAs. In patients with primary THAs there were no dislocations; however, one of the four postoperative deaths occurred following primary THA. We followed 75 hips for 7 (2-21) years; 51 patients had died by the time of the study. Neurological status deteriorated over time with 57% of patients progressing to functional stages IV or V, although consistent improvement was noted for pain relief. Function was directly related to the stage of the neurological disease. PMID- 12078879 TI - Total hip replacement with second generation cementing technique and the monobloc ScanHip: a 10-year follow-up. AB - In 230 consecutive patients 244 hips were operated with a 32-mm cemented prosthesis. The mean age at surgery was 77 (46-96) years. At the 10-year follow up 89 patients (95 hips) were dead and 33 patients (35 hips) did not attend because of old age or medical problems. Thirteen hips had been revised for aseptic loosening, two for infection, one for recurrent dislocations, and one for fracture. Four hips were found to be loose and seven had suspected radiographic loosening zones but only minor clinical symptoms. No or little limp was experienced by 52% of patients and 62% reported no or little pain. Eighty-seven percent were satisfied with the early postoperative result and 77% were still content at the time of follow-up. On the 10-year radiographs wear of the cups was, on average, 2.2 mm in loose hips and 1.5 mm in intact hips (P=0.02). There was no correlation between dissatisfaction and loosening visible by X-ray at follow-up. PMID- 12078880 TI - Press-fit stability of uncemented hemispheric acetabular components: a comparison of three porous coating systems. AB - Cementless acetabular components require good initial fixation to allow bony in growth. The initial press-fit stability, important for designs that do not rely on supplemental fixation, was examined for three designs with different porous coating systems: beads, fiber mesh, and plasma spray. The ability to withstand tangential loads was determined (maximum rim loads: 122-1730 N). The plasma sprayed acetabular cups withstood the greatest tangential load within the specified range of motion of 150 microm (P<0.01). Differences in surface preparation of titanium acetabular cups may significantly affect the initial stability of the implants in rim loading. PMID- 12078881 TI - Internal hemipelvectomy and reconstruction with a megaprosthesis. AB - We treated nine consecutive patients by internal hemipelvectomy and reconstruction with custom-made megaprosthesis between 1990 and 1997. Four had a primary malignant bone or soft tissue tumour, one a recurrent benign giant cell tumour, three solitary metastatic bony lesions, and one a pelvic defect secondary to multiple revision procedures after total hip replacement. After a mean follow up period of 62 (40-102) months five patients were still alive, including four of the eight patients with tumours. Three patients died as a result of the malignant disease, and one died of an infection related to the surgical treatment after 10 41 months. In two patients removal of the prosthesis was required due to infection, and six patients suffered various complications. One patient had an excellent outcome. PMID- 12078882 TI - Exchange nailing for aseptic nonunion of the femoral shaft. AB - Thirty-six nonunions of the femoral shaft were treated by closed removal of previous nails, reaming of marrow cavity as widely as possible, and reinsertion of stable intramedullary nail. Indications included femoral shaft aseptic nonunions with inserted intramedullary nail, nonunions with a radiolucent line, without segmental bony defects, and less than 1 cm shortening. They were followed up for a median of 2.9 (1-6) years. Thirty-three nonunions healed uneventfully within a median period of 4 (3-8) months. Except for three persistent nonunions, no other significant complications occurred. PMID- 12078884 TI - Total knee arthroplasty in patients with severe haemophilia. AB - We reviewed the perioperative records of haemophiliac patients who underwent total knee replacement (TKR) over the period 1990-1999 and followed up the surviving patients. Nine knees were replaced in eight patients. Seven of the nine patients did not require blood transfusion. Mean perioperative haemoglobin drop was 2.9 g/dl. There were no significant complications. All surviving patients (eight TKRs) were reviewed clinically using a standard scoring system. The mean follow-up period was 51 months. All patients scored excellent (>90/100) in the functional category. The frequency of haemarthroses fell significantly from the rate experienced prior to TKR, and pain scores were zero. PMID- 12078883 TI - Second-generation intramedullary supracondylar nail for distal femoral fractures. AB - The objective of this study was to review the use of intramedullary supracondylar (IMSC) nails for distal femoral fractures. We reviewed 24 fractures treated with second-generation IMSC nails. The fractures consisted of 18 type A1, one type A2, two type C1, one type C2, and two type C3 fractures. The relationships between clinical results and fracture type, approaches, and patient age were retrospectively reviewed. All fractures healed clinically and radiographically. Twenty-one patients maintained gait performance equivalent to that before injury. Average operating time was 108 min +/- 43 min. ROM in the knee of all patients was -5 degrees +/- 6 degrees in extension and 102 degrees +/- 38 degrees in flexion. Extension lag was influenced by surgical approach. The final knee arc was inversely correlated to patient age (R: 0.49, P<0.05). There were three varus/valgus deformities, two cases with loosening, and two with breakage of the distal locking screws, but no failure of the nail itself. Second-generation IMSC nailing for distal femur fractures was satisfactory in patients younger than 60 years of age. PMID- 12078885 TI - Dual Articular Knee in demanding primary and revision replacements in patients with rheumatic diseases. AB - We report the results of 25 total knee replacements in 24 patients with rheumatoid arthritis (RA) using the Dual Articular Knee prosthesis. There were four primary and 21 revision procedures. The main indication was severe joint instability. In four infected arthroplasties a two-stage revision procedure was used. Four patellar tendon avulsions and one deep infection were encountered. Results were excellent in 18 patients. Dual Articular Knee proved to be favourable in both demanding primary as well as revision arthroplasties in patients with RA. PMID- 12078886 TI - Long-term results of arthroscopic synovectomy for seropositive rheumatoid arthritis: 6-16 year review. AB - We prospectively studied a consecutive series of 25 knees (21 patients) treated with arthroscopic synovectomy for seropositive rheumatoid arthritis. All patients had pain and swelling and were in the early stages of the disease process (Larsen grade 2 or less). Three patients were lost to follow-up. At a mean of 8 years from operation two knees underwent total knee replacement with another two knees required a further arthroscopic synovectomy. One patient continued to experience intermittent mild synovitis. The range of movement was maintained or improved by surgery in 73% of cases but radiological evidence of degenerative change was seen in all knees. We discuss the technical difficulties associated with arthroscopic synovectomy that were associated with a small complication rate. In appropriately selected patients unresponsive to medical therapy, arthroscopic synovectomy can give safe and reliable results. PMID- 12078887 TI - An overview of the Fifth International Symposium on Hodgkin's lymphoma: recent advances in basic and clinical research. PMID- 12078888 TI - ABVD versus stanford V versus MEC in unfavourable Hodgkin's lymphoma: results of a randomised trial. AB - BACKGROUND: Between January 1996 and April 2000, 355 patients with advanced Hodgkin's disease (HD) (stage II bulky disease, III and IV) were enrolled in a prospective, multicentre, randomised trial aimed at comparing the efficacy of two new promising regimens: Stanford V and MEC hybrid. ABVD was chosen as the control arm. Radiotherapy was planned at the end of induction therapy on residual masses or on sites of previous bulky lesions. One hundred and seventeen, 123 and 115 patients were treated with Stanford V, MEC and ABVD, respectively. The records of 275 enrolled patients (89 Stanford V, 88 MEC, 98 ABVD) have been reviewed and are the subject of this report. RESULTS: After induction therapy a complete response (CR) was observed in 93, 89 and 74% of patients treated with MEC, ABVD and Stanford V, respectively, with a statistically significant difference (P = 0.013) between the arms. After a median follow-up of 24 months, 16 relapses have been recorded among 196 patients who achieved a CR. Relapse rates are 16, 6 and 4% for Stanford V, ABVD and MEC, respectively (P = 0.042). The 3-year survival was 93%, without any significant difference among the arms. However, a significant difference emerged in terms of failure free survival (FFS). Patients treated with Stanford V did the worst compared with those treated with ABVD or MEC (P = 0.001). Toxicity was comparable in the three treatment arms. CONCLUSION: For this randomised study, both ABVD and MEC gave superior results to Stanford V in terms of response and FFS; MEC seems to be the best regimen in terms of relapse-free survival, even if a significant difference has not yet been achieved. Notwithstanding the short follow-up, these results seem to be very impressive in defining the best standard treatment for HD for this subset of patients. PMID- 12078889 TI - Feasibility of upfront dose-intensive chemotherapy in children with advanced stage Hodgkin's lymphoma: preliminary results from the Children's Cancer Group Study CCG-59704. AB - BACKGROUND: Treatment strategies involving dose intensification have recently demonstrated improvements in cure compared with older trials. However, dose intensive therapy is associated with increased acute and long-term toxicities, particularly in pediatric patients. The Children's Cancer Group initiated this pilot study to assess the feasibility and toxicity of a moderate dose-intensive regimen, BEACOPP (bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine and prednisone), in children and adolescents with advanced-stage Hodgkin's lymphoma (HL). PATIENTS AND METHODS: Children with stage IIB or IIIB with bulk disease, or stage IV were eligible. Induction consisted of four cycles of escalated dose BEACOPP. The rapidity of response, defined as >70% reduction in disease burden, was assessed after two and four cycles. Rapid responders then received consolidation therapy as per gender-specific guidelines to reduce the risk of gender-specific long-term toxicities of therapy, i.e. females received four cycles of COPP/ABV (cyclophosphamide, vincristine, procarbazine, prednisone, doxorubicin, bleomycin and vinblastine) without radiation therapy and males received two cycles of ABVD (doxurubicin, bleomycin, vinblastine and dacarbazine) with involved field radiation therapy (IFRT). Slow responders received four cycles of BEACOPP and IFRT. RESULTS: Ninety-nine patients were enrolled. Myelosuppression was frequent. Non-hematological grade 4 toxicities included allergic reaction (two patients), hypotension (one), mucositis (four), infection (three), seizure (one) and elevated transaminases (one). Typhlitis developed in four patients; three recovered and completed dose modified chemotherapy, while one died of sepsis associated with grade 4 neutropenia. A rapid response was achieved by 45 and 72% of patients after two and four cycles, respectively. There are no disease progressions or secondary malignancies to date. There is only one reported relapse to date. Median follow up for the cohort is 6 months. CONCLUSIONS: BEACOPP chemotherapy is feasible and generally well tolerated in children with advanced-stage HL. The absence of reported progressive disease and only one relapse to date is encouraging. PMID- 12078891 TI - Prognostic factors and treatment outcome in patients with primary progressive and relapsed Hodgkin's disease. PMID- 12078890 TI - Biology of Hodgkin's lymphoma. AB - Significant progress has been made in recent years in our understanding of the cellular origin of Hodgkin and Reed-Sternberg (HRS) cells in Hodgkin's lymphoma (HL). It is now clear that in most instances HRS cells represent clonal populations of transformed germinal centre (GC) B cells. While the tumour cells in the lymphocyte predominant type of the disease resemble mutating and antigen selected GC B cells, there is evidence that HRS cells in classical HL originate from pre-apoptotic GC B cells. HRS cells of the recently defined novel subtype lymphocyte-rich classical HL moleculary resemble HRS cells of the other types of classical HL, but there appear to be phenotypic differences. In rare cases, HRS cells derive from T cells. In contrast to previous speculations, cell fusion apparently does not play a role in the generation of the tumour clone. By gene expression profiling of HL cell lines, it became evident that HRS cells have lost most of the B cell-typical gene expression program, which may explain why these cells can persist without B cell receptor expression and which suggests that at least one of the transforming events involved in HL pathogenesis affects a master regulator of cell lineage identity. PMID- 12078893 TI - Autologous stem cell transplantation in Hodgkin's disease. PMID- 12078892 TI - New concepts for relapsed Hodgkin's disease. PMID- 12078894 TI - Role of allogeneic stem cell transplantation in relapsed or refractory Hodgkin's disease. AB - Little information is available regarding allogeneic stem cell transplantation (alloSCT) and Hodgkin's disease (HD). Autologous stem cell transplantation (autoSCT) is usually preferred to alloSCT due to its widespread availability, lack of the immunological problems intrinsic to the development of graft-versus host disease (GvHD), and the infrequent bone marrow involvement present in HD patients undergoing high-dose chemotherapy/radiotherapy. AlloSCT has been associated with a high transplant-related mortality (TRM) in patients with HD due to a high incidence of GvHD and of fatal infectious events after transplantation. The poor outcome of these patients after alloSCT may reflect in part the advanced status of the disease at transplantation and the poor performance status of the patient population allografted. Furthermore, the high TRM present in t h e conventional alloSCT setting hasnever allowed a proper evaluation of a possible graft-versus-Hodgkin's effect. In an effort to reduce the TRM associated with alloSCT, low-intensity regimens have been developed; the curative potential of these protocols would rely on the graft-versus-leukemia effect of the allogeneic infusion more than in the conditioning regimen per se. Although the number of HD patients allografted with reduced-intensity protocols is low and the follow-up still short, TRM seems lower than in the conventional allograft setting despite a similar incidence of acute GvHD (aGvHD). Overall and progression-free survival seem promising, and patients developing aGvHD after transplantation or donor lymphocyte infusions seem to be at a lower risk of relapse than those not presenting this complication. PMID- 12078895 TI - Hodgkin's disease in the elderly: current status and future directions. AB - In general, it was agreed that high rates of toxicities during treatment occur in the elderly and that there is a frequent occurrence of early relapse. It is clear that different combinations of effective therapies with lower toxicity are required. It was felt, however, that certainly in the 60-70 year age group, approaches should be vigorous to and the same diagnostic and staging procedures as in younger individuals, but with much closer monitoring of toxicity and response to treatment. It was felt that as part of the approach, liberal support with haemopoeitic growth factors (G-CSF) was necessary to reduce prolonged neutropenia. It is important to understand that age in general is not a contrary indication for aggressive treatment and that biologically younger patients under the age of 65 years, in good physical and mental condition, often should be given with stage-adapted treatment, analogous to conventional treatment protocols for the <60 years age group. It was also considered that, in patients who clearly could not accept conventional treatment, study groups could begin to define the best palliative care for patients with pre-existing organ impairment, and that in all situations of assessment, whether in trial or not, there should be a detailed prospective assessment of quality of life parameters. PMID- 12078896 TI - Fertility after treatment for Hodgkin's disease. AB - BACKGROUND: The investigational endeavors of ovarian cryopreservation await the clinical experience of auto- or xenotransplantation, in vitro maturation of thawed primordial follicles, their in vitro fertilization and embryo transfer. Although promising, this experience is not yet available. Moreover, the risk of possible reimplantation of malignant stem cells with the thawed cryoperserved ovary has been raised following experimental animal observations. Therefore, until these innovative endeavors prove successful, we have attempted to minimize the gonadotoxic effect of chemotherapy by the co-treatment with a gonadotropin releasing hormone agonistic analog (GnRH-a) to induce a temporary prepubertal milieu. The immunoreactive inhibin-A and -B in these patients was measured before, during and following the gonadotoxic chemotherapy. METHODS: A prospective clinical protocol was undertaken in 60 women aged 15-40 years with lymphoma, 10 with leukemia and 10 undergoing chemotherapeutic treatments for non-malignant diseases such as systemic lupus erythematosus or other autoimmune diseases. A monthly injection of depot D-TRP(6)-GnRH-a was administered from before starting the chemotherapy until its conclusion, up to a maximum of 6 months. Hormonal profile [follicle-stimulating hormone (FSH), luteinizing hormone (LH), E2, T, P4, insulin-like growth factor (IGF)-1, IGF-BP3 and prolactin) was taken before starting the GnRH-a/chemotherapy co-treatment, and monthly thereafter until resumtion of spontaneous ovulation. This group was compared with a control group of 60 women who have been treated with similar chemotherapy. RESULTS: Whereas all but three (40, 36 and 34 year old) of the surviving patients within the GnRH-a/ chemotherapy co-treatment group resumed spontaneous ovulation and menses within 12 months, less than half of the patients in the 'control' group (chemotherapy without GnRH-a co-treatment) resumed ovarian function and regular cyclic activity (P <0.05). The remaining 55% experienced premature ovarian failure (POF). Temporarily increased FSH concentrations were experienced by about one-third of the patients resuming cyclic ovarian function, suggesting reversible ovarian damage in a larger proportion of women than those experiencing POF. Inhibin-A and -B decreased during the GnRH-a/ chemotherapy co-treatment but increased to normal levels in patients who resumed regular ovarian cyclicity, and/or spontaneously conceived, as compared with low levels in those who developed POF. CONCLUSIONS: If these preliminary data are consisent in a larger group of patients, GnRH-a co treatment should be considered in every woman of reproductive age receiving chemotherapy, in addition to assisted reproductive technologies and the investigation into ovarian cryopreservation for future in vitro maturation, autotransplantation or xenotransplantation. PMID- 12078897 TI - Genetic instability in Hodgkin's lymphoma. AB - Genetic instability is a characteristic feature of the malignant Hodgkin and Reed Sternberg (HRS) cells in classical Hodgkin's lymphoma and the lymphocytic and histiocytic (L&H) cells in lymphocyte predominant Hodgkin's lymphoma. Genetic instability can be classified into four major categories: distinct DNA mutations (microsatellite instability); numerical aberrations (chromosomal instability); structural aberrations (translocation instability); and gains and losses of chromosomal regions. In Hodgkin's lymphoma (HL), HRS cells and L&H cells show somatically mutated clonally rearranged immunoglobulin genes, thus characterizing these cells genetically as germinal center B cells. These cells furthermore show mutations of oncogenes and tumor suppressor genes in some cases (p53, IkappaBalpha, CD95/Fas). They do not, however, display microsatellite instability, as they have a proficient mismatch repair machinery. In contrast, HRS and L&H cells frequently harbor recurrent but not specific numerical and structural aberrations as detected by classical cytogenetics and fluorescence in situ hybridization analysis. Results from molecular genetic studies using comparative genomic hybridization and allelotyping (LOH) indicate typical genetic patterns in HL with gains and losses of distinct chromosomal regions. In some instances, candidate genes possibly involved in the malignant transformation of HRS cells and L&H cells have been characterized (JAK2, c-REL, MDM2). In summary, using molecular genetics it might be possible in the near future to elucidate some of the complex genetic instabilities observed in HL. PMID- 12078898 TI - Viruses and Hodgkin's lymphoma. AB - Hodgkin's lymphoma (HL) is unusual among human malignancies in that the epidemiology suggests an infectious aetiology. The Epstein-Barr virus (EBV) is associated with a proportion of cases and this association is believed to be causal. In these cases the Hodgkin and Reed-Sternberg (HRS) cells express the EBV encoded proteins LMP1 and LMP2, which can mimic CD40 and the B cell receptor, respectively, and therefore may play a critical role in facilitating the survival of HRS cells. EBV-associated and non-EBV-associated HL cases have different epidemiological features and recent data suggest that delayed exposure to EBV is a risk factor for the development of EBV-associated HL in young adults. We suggest that HL can be divided into four entities on the basis of EBV status and age at presentation, with three groups of EBV-associated cases and a single group of EBV-negative cases. The aetiology of the latter cases is obscure although involvement of an infectious agent(s) is suspected. PMID- 12078899 TI - Genetic susceptibility to Hodgkin's lymphoma and to secondary cancer: workshop report. AB - Although the occurrence of familial Hodgkin's lymphoma (HL) is a rare event, genetic susceptibility as a cause of HL and its influence on treatment outcome may not be rare. However, results obtained from the analysis of HL families will probably have broad implications with regard to understanding common pathogenic factors leading to the development of the disease. The description of anticipation among the affected offspring of HL patients further strengthens the view that heritable factors contribute to development of HL. Moreover, the finding that particular human leukocyte antigen (HLA) alleles are associated with susceptibility to HL may be regarded as a hint to the presence of an as yet undefined infectious agent, leading to the growth of a malignant lymphoma cell clone in those patients that are more susceptible to this agent due to their HLA genotype. In addition, since an intrinsic genomic instability was observed in a proportion of HL patients, it is plausible that these patients are not only susceptible to the causation of HL, but are also at a higher risk of developing therapy-related (TR) secondary cancers following treatment. Estimation of sister chromatid exchange was established as a tool to identify patients at higher risk of TR cancer. In this context the use of therapeutic agents known to increase genomic instability should be carefully considered prior to determing the best treatment. The future identification of heritable factors contributing to HL will be of importance both with regard to diagnosis as well as treatment of HL patients. PMID- 12078900 TI - Polymorphisms of drug metabolizing enzymes and markers of genotoxicity to identify patients with Hodgkin's lymphoma at risk of treatment-related complications. AB - Survivors of childhood Hodgkin's lymphoma (HL) have an increased risk of developing treatment-related complications, especially second malignant neoplasms, as a result of treatment regimens incorporating chemotherapy and radiation therapy. Second cancers include leukemias that generally occur in the first two decades after therapy, and adult-type solid tumors that generally exhibit continued increasing incidence throughout subsequent follow-up. Identified clinical risk factors for second cancers include age at the time of treatment and intensity and type of therapy, with particularly strong associations between the use of radiotherapy and subsequent breast cancer, and alkylator chemotherapy dose-intensity and risk of secondary leukemia. However, second cancers affect a minority of patients, and there is probably great variability in individual susceptibility for this complication. Common genetic polymorphisms in drug-metabolizing enzymes that result in impaired detoxification of chemotherapy or inefficient repair of drug- or radiation-induced genetic damage may lead to increased risk of a second cancer. Studies of the potential role of polymorphisms in the genes encoding the glutathione S-transferases, cytochrome P450 3A4, NAD(P)H:quinone oxidoreductase and myeloperoxidase in the etiology of treatment-related complications are reviewed. Biological markers of drug- and radiation-induced genetic damage may also identify patients at higher risk of immediate and delayed side effects of therapy. The Children's Oncology Group (COG) is examining the roles of polymorphisms in drug metabolizing enzymes and biological markers of genotoxicity in predicting the treatment-related outcomes of patients with HL. These investigations may ultimately allow the use of pharmacogenetically guided therapy to improve the outcome of HL therapy and reduce the risk of therapy-related complications, especially secondary malignancies. PMID- 12078901 TI - Genetic susceptibility to Hodgkin's disease and secondary neoplasias: FISH analysis reveals patients at high risk of developing secondary neoplasia. AB - BACKGROUND: Cytotoxic drugs administered before high-dose therapy (HDT) represent a significant factor in the development of leukemic complications in patients with lymphoid malignancies. This retrospective study was used to detect evidence of abnormal therapy-related myelodysplasia/secondary acute myeloid leukaemia (tMDS/sAML) clones before HDT in a subset of patients who subsequently developed secondary neoplasia. PATIENTS AND METHODS: 230 patients with non-Hodgkin's lymphoma (NHL) underwent HDT comprising cyclophosphamide and total body irradiation (TBI) with autologous hematopoietic progenitor-cell support. Thirty three patients have developed tMDS/sAML and 20 of these were screened for the presence of emerging therapy-related abnormalities before HDT. A further 24 patients without evidence of secondary neoplasia were screened using fluorescence in situ hybridisation (FISH). RESULTS: Significant levels of abnormal cells were identified in 20/20 patients screened who have developed secondary neoplasia compared with only three of 24 patients in the HDT control group who have not. The latter three patients have since died. CONCLUSIONS: The triple FISH assay was developed to detect loss of chromosomal material from 5q31, 7q22 and 13q14. It can potentially identify those patients at risk of alkylating agent-induced leukaemia before they proceed to HDT. Used in a prospective manner, the triple FISH assay could permit more informed clinical management. PMID- 12078903 TI - Late mortality in Hodgkin's disease: can we reduce it? PMID- 12078902 TI - Workshop on the relationship between nodular lymphocyte predominant Hodgkin's lymphoma and T cell/histiocyte-rich B cell lymphoma. PMID- 12078904 TI - Chemokines, cytokines and their receptors in Hodgkin's lymphoma cell lines and tissues. AB - BACKGROUND: Hodgkin's lymphoma (HL) is characterized by a minority of neoplastic cells, the so-called Reed-Sternberg (RS) cells, and an admixture of reactive cells including lymphocytes, plasma cells, eosinophils and histiocytes. Cytokines produced in HL, either by RS or infiltrating cells, might explain the presence and maintenance of an impaired immune response. Chemokines (cytokines with chemoattractant properties) produced by RS cells play a major role in leukocyte trafficking. These molecules with specific receptor affinities contribute, for example by attracting TH2-like T cells, to the maintenance of a favorable environment for survival of RS cells. RESULTS AND DISCUSSION: Cross-talk between RS cells and reactive elements involves several cytokine/chemokines that process proliferative [interleukin (IL)-13 and IL-17], immunosuppressive (IL-10 and transforming growth factor-beta) and background formation (IL-5, TARC, MDC, IP 10, RANTES, Mig and others) messages. These autocrine and paracrine interactions lead to an environment where RS cells are able to proliferate, escape from apoptosis and survive host anti-tumor defense. CONCLUSIONS: The proper modulation of these complex pathways may allow the development of new strategies in HL therapy. PMID- 12078905 TI - Current strategies of antibody-based treatment in Hodgkin's disease. AB - Many new approaches involving antibody-based agents have given promising results in experimental Hodgkin's disease (HD) models. Clinical trials with monoclonal antibodies, immunotoxins, bispecific constructs and radioimmunoconjugates have demonstrated some clinical efficacy in patients with advanced refractory HD. Although it seems unlikely that resistant patients with larger tumor masses will be cured by either of these approaches, it might be feasible to treat bulky disease by conventional therapy and then administer biological agents to kill residual Hodgkin and Reed-Sternberg cells. Future phase III trials will have to prove a possible superior effect of this combined immunochemotherapy. Currently, the evaluation of the most promising approaches continues. PMID- 12078906 TI - Prognostic factors in Hodgkin's lymphoma. AB - Risk-adapted treatment strategies have constituted a major issue since the beginning of clinical research into Hodgkin's disease (HD). Various prognostic factors have been identified and several of those considered for staging procedures, resulting in strictly stage-dependent treatment recommendations for patients suffering from HD. These factors may be subdivided in host-related (e.g. age, sex) and tumour-related (e.g. number of tumour cells, growth characteristics, spread of tumour cells, resistance to apoptosis) factors. Owing to the striking improvement of the overall prognosis in HD patients it may be difficult to identify novel prognostic factors analysing the minority of patients with a fatal outcome. However, especially in advanced-stage disease, improved treatment results were achieved by the introduction of more aggressive treatment regimens, resulting in an increased toxicity rate. Thus, partially in contrast to earlier work in this field, future prognostic factors are needed for identification of those patients that have a good prognosis and might be susceptible to overtreatment. During the Fifth International Symposium on Hodgkin's Lymphoma, promising results on several new prognostic markers were presented. Furthermore, a joint effort to design new studies on large, well characterised patient groups has been initiated. PMID- 12078907 TI - The disappearance of prognostic factors in Hodgkin's disease. AB - The purpose of this conceptual (and somewhat provocative) article is to analyse the reasons for the disappearance of prognostic factors in Hodgkin's disease (HD) and to explore the consequences for further research from a biometrical point of view. The very concept of prognostic factors is about to resolve in HD. Prognostic factors gradually loose their predictive power as treatment is successfully adapted to the disease burden. Freedom from treatment failure and overall survival curves of patients in early, intermediate and advanced stages of HD are superimposable with the best current treatment protocols of the German Hodgkin's Lymphoma Study Group. This disappearance of prognostic factors in HD necessitates a certain conceptual remodelling. It's time to think quantitatively and bivariately, and we need to (i) synthesise existing 'prognostic' factors into a quantitative measure of disease burden or severity; (ii) develop a quantitative measure of treatment strength and (iii) relate these two quantities in nomogram curves indicating how much treatment a patient with a given disease burden requires to have, say, a 85%, 90% and perhaps 95% expected chance of cure. PMID- 12078908 TI - The involved field is back: issues in delineating the radiation field in Hodgkin's disease. AB - During the last century, the role of radiation therapy in the treatment of Hodgkin's disease (HD) has changed drastically. From a palliative treatment reserved for bulky lymph nodes of an incurable disease at the beginning of the century, to an exciting primary treatment used alone to cure most stages in the 1960s and 1970s, to the present more limited role as consolidation treatment after chemotherapy. Interestingly, the radiation field size has always influenced the evolution of treatment principles of HD. Over several decades, large or extended field radiotherapy has become synonymous with the successful treatment of HD. But the critical transformation from a single-modality to a combined modality therapy, together with improvement in imaging and radiation planning techniques, mandates a reassessment of the delineation of appropriate radiation fields in HD. In this manuscript we review the comeback of the involved field, address design questions and offer field borders for common disease sites. PMID- 12078909 TI - Treatment of early and intermediate stage Hodgkin's lymphoma in the German Hodgkin's Lymphoma Study Group. PMID- 12078911 TI - Current clinical trials for advanced Hodgkin's lymphoma in North America: history, design and rationale. PMID- 12078910 TI - Early response to chemotherapy: a surrogate for final outcome of Hodgkin's disease patients that should influence initial treatment length and intensity? AB - BACKGROUND: Early adjustment of treatment may benefit the patient. In order to guide treatment adjustment, use of early response (ER) or early complete response (ECR), judged after the few initial cycles of chemotherapy, is common in pediatric and also adult Hodgkin's and non-Hodgkin's studies. Paradoxically, almost no data support this strategy. PATIENTS AND METHODS: The influence of ECR on outcome was evaluated in three series of advanced Hodgkin's disease (HD), leading to a series of questions. RESULTS: The 1982 EORTC study assessed prospectively the time frame needed to reach an apparent complete response (CR) through repeated tumor measurements. In patients assessed at mid-treatment before the fifth cycle, both 15 year freedom from progression (FFP) and overall survival (OS) were superior in ECR patients compared with other patients continued on the same treatment (61% versus 37%; P < 0.001). A series of questions arise from these observations. Question 1: is the shortening of treatment detrimental? In a randomized Swedish trial, in one arm treatment was shortened in patients evaluated from the fifth cycle as ECR as compared with the standard eight cycles arm, 10 year cause-specific-survival (CSS) was 53 versus 69% [not significant (ns)]; 10 year OS 49% versus 58% (ns). Conversely, in the EORTC 20884 study, ECR patients given only six cycles did as well as patients entering CR later and, for this reason, given eight cycles (identical 6 year event-free survival 75%). Question 2: is early treatment adaptation in patients who failed to reach ER beneficial? In the French MDH 90 trial, 15% of children failed to reach ECR after four cycles; in these children only, anthracyclines plus alkylating agents were given and the dose of radiotherapy increased, improving the results observed in the previous trial. In the EORTC 20884 study, patients who failed to reach an ECR were switched earlier to involved field RT: their results matched those of ECR patients, at the difference of the previous trial. Question 3: is ER a predicting factor that can be used with any type of treatment? Probably not, based on the German Hodgkin's Lymphoma Study Group trial HD 9: ECR is highly dependent on specific interval from treatment start and on treatment intensity. DISCUSSION: More general questions stem from these results. Question 4: is the definition of ER secured? With conventional imaging, the different methods for response assessment at end treatment also lead to different response rates; the assessment in the middle of treatment itself and the use of newer imaging techniques may further increase the variation. Indeed, question 5 is: is ER a concept based on any biology? Correlation to markers, 99mTc uptake, PET and hematological tolerance might help to pinpoint how and why ER represents a surrogate for final outcome. CONCLUSION: ER is a surrogate for final outcome, reflecting both tumor burden and activity. This predictability may, and possibly should, impact on treatment. PMID- 12078912 TI - Current clinical trials for the treatment of adult advanced-stage Hodgkin's disease: GELA experiences. Groupe d'Etudes des Lymphomes de l'Adulte. AB - BACKGROUND: The optimal treatment for patients with advanced Hodgkin's disease (HD) responding to initial chemotherapy (CT) and an intensive salvage therapy for those who fail to respond completely after initial treatment were evaluated prospectively. PATIENTS AND METHODS: The Groupe d'etudes des Lymphomes de l'Adulte H89 trial compared two cycles of CT with (sub)total nodal irradiation (RT) as consolidation treatments for patients with stage IIIB/IV HD with a complete response (CR) or good partial response (PR) after six cycles of CT. Early salvage therapy, including intensified cytoreductive CT and high-dose CT with autologous stem-cell transplantation, was integrated into the trial for patients who had failed to respond completely or relapsed after initial treatment. RESULTS: The study does not demonstrate any advantage of RT over CT as consolidation treatment at the time of CT-induced CR or good PR. Early intensive therapy improves the outcomes of patients with PR and those who relapsed with unfavourable factors. This strategy remains unsatisfactory for patients with primary refractory disease and chemoresistant disease. CONCLUSION: Based on first intensification of conventional-dose CT, in the next trial (EORTC-GELA Intergroup Study), four escalated bleomycin-etoposide-doxorubicin-cyclophosphamide procarbazine-prednisone (BEACOPP) followed by four baseline BEACOPP are compared with the eight doxorubicin-bleomycin-vinblastine-dacarbazine standard with no RT for patients who achieve CR/CR-uncertain after initial CT. PMID- 12078913 TI - Current clinical trials for the treatment of advanced-stage Hodgkin's disease: BEACOPP. AB - BACKGROUND: The bleomycin-etoposide-doxorubicin-cyclophosphamide-vincristine procarbazine-prednisone (BEACOPP) regimen was developed to investigate the potential of moderate dose escalation of conventional polychemotherapy to improve the unsatisfactory treatment results in advanced-stage Hodgkin's lymphoma (HL). Following pilot studies, the randomised trial HD 9 demonstrated that BEACOPP (baseline dose) attained superior failure-free survival to COPP/ABVD, and that dose escalation made a further marked improvement. Toxicity was severe but manageable. PATIENTS AND METHODS: The current GHSG multicentre randomised trial HD 12 has a 2 x 2 factorial design in order to make two comparisons: (i) eight cycles of escalated BEACOPP (as in HD9) are compared with four escalated cycles followed by four at baseline dose; (ii) the use of additional local radiotherapy to initial bulky disease and residual disease after chemotherapy is compared with chemotherapy alone, except where radiotherapy was prescribed by a central diagnostic panel. Eligible are patients aged 16-65 years with newly diagnosed HL of stage IIB with risk factors or stage III/IV. The EORTC multicentre trial 20012 randomises patients with HL stage III/IV to either eight cycles of ABVD or eight cycles of BEACOPP (four escalated + four baseline). RESULTS: The first interim analysis (January 2001) of HD 12 with 221 evaluable patients indicated continuation of recruitment. Recruitment will end in 2002 and the final data analysis will appear in 2006. CONCLUSIONS: The BEACOPP regimen is highly effective, and moderate dose escalation makes a further worthwhile improvement in tumour control. Current trials will measure BEACOPP against the international standard and show whether the amount of chemotherapy and/or radiotherapy can be reduced. PMID- 12078914 TI - Carbohydrate and satiety. AB - This review focuses on what is known about the effects of carbohydrate on food intake, the potential mechanisms mediating these effects, and the impact of different monosaccharides in humans. The inhibition of subsequent food intake associated with ingestion of carbohydrate appears to result primarily from gastrointestinal signals, including those generated by orosensory stimulation, gastric distension, and perhaps most importantly the interaction of nutrients with receptors in the small intestine. The latter is associated with the release of putative satiety hormones, including glucagon-like peptide-1 and amylin, and slowing of both gastric emptying and small intestinal transit (thereby prolonging gastric distension and increasing the time available for nutrient absorption). The effects of carbohydrate on food intake are dependent on the route of administration (i.e., oral, intragastric, or intraduodenal). Changes in blood glucose and insulin concentrations per se probably do not play a major role in the induction of satiety. Studies relating to the comparative effects of different monosaccharides/carbohydrates have yielded inconclusive results, probably in part owing to substantial differences in methodological approaches. PMID- 12078915 TI - Olive oil as a functional food: epidemiology and nutritional approaches. AB - Olive oil is an integral ingredient of the Mediterranean diet and accumulating evidence suggests that it may have health benefits that include reduction of risk factors of coronary heart disease, prevention of several varieties of cancers, and modification of immune and inflammatory responses. Olive oil appears to be an example of a functional food, with varied components that may contribute to its overall therapeutic characteristics. Olive oil is known for its high levels of monounsaturated fatty acids and is also a good source of phytochemicals including polyphenolic compounds, squalene, and alpha-tocopherol. PMID- 12078916 TI - Understanding copper uptake at the molecular level. AB - Despite significant advances in our understanding of copper efflux mechanisms since the discovery and characterization of Cu-ATPase genes mutated in Menkes and Wilson diseases, little is known about how cells acquire this essential micronutrient. Recent studies on Ctr1 have illuminated how copper may be transported into mammalian cells. PMID- 12078917 TI - Nature's perfect food revisited: recent insights on milk consumption and chronic disease risk. AB - In Swedish volunteers, consuming spaghetti along with milk raises the insulinemic effect of the meal. In Finnish citizens followed for 19 to 24 years for cancer incidence, different consumption patterns of dairy items had different relationships with colon and rectal carcinogenesis. More definitive and conclusive research into how dairy products might influence chronic disease would seem to be warranted. PMID- 12078918 TI - Is IL12B a potential genetic target for the prevention of type 1 diabetes through dietary intervention? AB - Type 1 diabetes mellitus (T1D) results from decreased insulin production. Its clinical expression is the result of an array of genetic and environmental factors. The IL12B gene could be one of the loci implicated in the genetic susceptibility of T1D. Thus, this genetic predisposition could be modulated by dietary factors. However, there is no consistency among populations in relation to relevance of this locus as a candidate gene for T1D. PMID- 12078919 TI - FDA panel conditionally approves DNA pap test without ACOG support. PMID- 12078920 TI - Disparities in hospital use, mortality rates for people with AIDS. PMID- 12078921 TI - Higher prevalence of stroke in Afro-Americans. PMID- 12078922 TI - Black-white infant mortality disparity persists. PMID- 12078923 TI - EMTALA basics: what medical professionals need to know. Emergency Medical Treatment and Active Labor Act. PMID- 12078924 TI - Addressing medical coding and billing part II: a strategy for achieving compliance. A risk management approach for reducing coding and billing errors. AB - Medical practice today, more than ever before, places greater demands on physicians to see more patients, provide more complex medical services and adhere to stricter regulatory rules, leaving little time for coding and billing. Yet, the need to adequately document medical records, appropriately apply billing codes and accurately charge insurers for medical services is essential to the medical practice's financial condition. Many physicians rely on office staff and billing companies to process their medical bills without ever reviewing the bills before they are submitted for payment. Some physicians may not be receiving the payment they deserve when they do not sufficiently oversee the medical practice's coding and billing patterns. This article emphasizes the importance of monitoring and auditing medical record documentation and coding application as a strategy for achieving compliance and reducing billing errors. When medical bills are submitted with missing and incorrect information, they may result in unpaid claims and loss of revenue to physicians. Addressing Medical Audits, Part I--A Strategy for Achieving Compliance--CMS, JCAHO, NCQA, published January 2002 in the Journal of the National Medical Association, stressed the importance of preparing the medical practice for audits. The article highlighted steps the medical practice can take to prepare for audits and presented examples of guidelines used by regulatory agencies to conduct both medical and financial audits. The Medicare Integrity Program was cited as an example of guidelines used by regulators to identify coding errors during an audit and deny payment to providers when improper billing occurs. For each denied claim, payments owed to the medical practice are are also denied. Health care is, no doubt, a costly endeavor for health care providers, consumers and insurers. The potential risk to physicians for improper billing may include loss of revenue, fraud investigations, financial sanction, disciplinary action and exclusion from participation in government programs. Part II of this article recommends an approach for assessing potential risk, preventing improper billing, and improving financial management of the medical practice. PMID- 12078925 TI - Myocardial infarction in sickle cell disease. AB - Vasculo-occlusive crisis with organ infarctions occur in sickle cell disease (SCD). However, heart infarction is not commonly reported. We reviewed 19 cases of documented myocardial infarction (MI) in SCD patients. The true incidence may be higher because the diagnosis was often made at autopsy and was overshadowed during life by other musculoskeletal symptoms. Electrocardiography is frequently unhelpful. Skeletal muscle enzymes confound serum cardiac enzyme interpretation. The mechanism of MI in SCD is not exactly known, as coronary angiography is usually normal. MI frequently occurs in association with hypoxia, cor pulmonale, anemia, sepsis, acidosis, and renal failure. The aim of this article is to increase awareness for this complication and to prompt prospective studies to look at treatment strategies for myocardial infarction in SCD. PMID- 12078927 TI - Analysis of the Black Racial Identity Attitude scale in a substance-abusing population. AB - Racial attitude characteristics of African Americans may have important implications for patient-therapist matching in substance abuse treatment. The Black Racial Identity Attitude Scale (BRIAS) is a questionnaire used to measure racial identity attitudes. This study tested the stability and internal consistency of the BRIAS in an African-American substance-abusing population. This is the first known test of the BRIAS in a clinical population. African American veterans (n = 53) were administered the BRIAS to test for stability over time and internal consistency. Initial analysis of the instrument revealed that a majority of items were not stable over time. Using the initial results, we removed 26 problematic items. Three modified scales remained, each having marginal test-retest reliability. Two of the modified scales had moderately adequate internal consistency, and the third was minimally adequate. We found that the BRIAS did not demonstrate sufficient internal consistency or stability in this population to adequately identify the constructs of the Nigrescence Racial Identity Development theory of William Cross, Jr. There is a growing recognition of the need to explore the extent of racial, ethnic, and cultural factors in substance abuse behavior and treatment; therefore, we recommend that further attention be directed toward developing an instrument to reliably and accurately measure identity constructs among African-American clinical subjects. PMID- 12078926 TI - Breast conservation is an effective option in Black, medically indigent patients. AB - The purpose of the study was to evaluate the efficacy of lumpectomy and postoperative radiotherapy in an African-American, medically indigent population. From 1980 through 1996, a retrospective chart review was undertaken of the patients treated with radiotherapy after lumpectomy at an inner city hospital, whose patients are primarily African American and uninsured. One hundred and one patients were treated with breast conservation during this time. Of those, 72 were African American and with invasive carcinoma. The data were analyzed using JMP IN (SAS Institute). The study found that African-American patients with invasive carcinoma had 95.2% local control at 5 years and 87.9% at 10 years. The disease-free survival was 84.6% at 5 years and 65.3% at 10 years. Patients that received less than 50 Gray to the tumor bed had inferior local control, disease free survival, and overall survival (p < 0.0001 for all three). The 5-year and 10 year local control for DCIS, in a limited number of patients, was 95.2%. We conclude that lumpectomy followed by radiotherapy is an effective treatment strategy in the high-risk population of African-American, medically indigent patients. The local control and disease-free survival compare favorably to published controls in this traditionally high-risk patient population. PMID- 12078928 TI - Urban poverty and infant-health disparities among African Americans and whites in Milwaukee. AB - This study examined neighborhood and infant health disparities between African American and white mothers in Milwaukee, Wisconsin. Census-block data were used for 1990 and Vital Statistics data were used for 1992 through 1994. African American mothers lived in less desirable, more segregated neighborhoods than white mothers did in 1990. African-American infant and neonatal mortality rates were twice those of whites (2.3 and 2.0, respectively), while African-American postneonatal mortality rates were three times that of whites (3.0). African American low and very low birth weight rates were more than twice those of whites (2.5 and 2.6, respectively). All African-American mothers were nearly eight times as likely as all white mothers to have inadequate prenatal care, whereas poor African-American mothers were three times as likely to have inadequate prenatal care as were poor white mothers. Public health experts and practitioners may want to consider the communities of minority patients to devise interventions suitable for addressing health disparities. PMID- 12078929 TI - Retrospective analysis of hiccups in patients at a community hospital from 1995 2000. AB - Hiccups are a physiologic phenomenon noted in animals and humans. There is little understanding of what makes hiccups occur and whether or not they have any productive purpose. A retrospective analysis of all patients seen in a community hospital over a 5 year period was conducted to see who is affected by hiccups, evaluate laboratory findings in people with hiccups, and to see what the currently accepted treatment is for hiccups. The vast majority of patients were male, older than 50 years of age, and with co-morbid conditions. Laboratory values appeared to be of little value in determining whether treatment interventions would be effective. Gastroenterology was the service most consulted and EGD the most common procedural intervention conducted, but with little success. No treatments showed a statistically significant effect. PMID- 12078930 TI - Domestic violence screening in the emergency department of an urban hospital. AB - Victims of domestic violence appeal to the health care system through emergency room visits for injuries related to violent episodes or for proxy care for other complaints. Screening for persons who are at high risk for violence or who are victims of violence has not been performed in emergency rooms when patients present for care, nor have all health care professionals been educated in the ways to ask the questions and assess the patients. The questions for identifying domestic violence victims have not been routinely asked on admission to the emergency department, and documentation of this information is not consistent. The purpose of this survey study was to identify the numbers and characteristics of adult victims of domestic violence who present to the emergency department of an urban community medical center during a 1 0-day period to estimate the extent of the domestic violence in the community served by the acute care facility. Findings demonstrated that emergency department staff had difficulty asking the questions, and the responsibility for the screening was relegated to the triage nurse. Questions were not asked of each adult presenting to the emergency department, and health care staff identified various reasons for their resistance. Although only 12% of persons were screened and only during the hours of 0700 through 1900, positive screens for physical abuse were found in 24.6% (n = 20) of the 81 women screened. Routine screening of all patients and sensitivity to the needs of those who have experienced domestic violence are integral to prevention and safety of those who are victimized. Injury prevention programs can then be instituted in the community with the collaborative efforts of local citizen groups and the health care facility. PMID- 12078931 TI - Professional motivation and career plan differences between African-American and Caucasian dental students: implications for improving workforce diversity. AB - Vast disparities in oral health status coupled with projected decreases in African Americans enrolling in and graduating from dental school have heightened concern about the underrepresentation of African Americans in the dental profession. The purpose of this study was to explore differences between African American and white American students regarding demographics, professional motivations, and career plans. African-American (n = 104) and white American (n = 226) dental students completed a biographical data survey instrument, which included information about family background and professional motivations and plans, and rated descriptions of three practice arrangements. African-American students were more motivated to become a dentist to serve the public, plan to specialize, work in an urban area, and work part-time. White American students were more motivated to become a dentist based on factors related to family commitments. Race was a significant predictor for student ratings for both solo and employee practice. Study results have implications for health professions educators, administrators, and policy makers in their efforts to improve the recruitment and retention of African-American students, shape dental curricula to meet diverse student needs, and implement loan forgiveness programs to enhance minority student recruitment. PMID- 12078932 TI - Skin tumors in aging Long Evans rats. AB - We report 25 cases of skin neoplasm observed among 30 Long Evans rats serving as controls in a psychosocial behavioral study conducted in the Vivarium at Charles R. Drew University, Los Angeles, CA. The animals were 10 weeks old at the beginning of the study. All the skin tumors developed at 18 to 26 months of age and slowly enlarged over a period of 9 months. Multiple nodules occurred in 8 males and 6 females. None of the tumors regressed. The tumors were located around the hind leg and dorso-medial area and measured 1 to 2 cm. Physical examination revealed firm well demarcated dermal masses. Most of the tumor nodules were intradermal, and some had a central ulcerated or keratin-filled core. Microscopic examination performed on some of the tumors showed findings of classic Keratoacanthoma, whereas others showed histologic features suggestive of squamous cell carcinoma. These findings indicate a high rate (83%) of spontaneous skin neoplasms among aging Long Evans rats. To our knowledge, such a high rate of skin neoplasms in aged rodents has not been described in the literature. Furthermore, further studies should be undertaken to confirm these findings and to assess whether these rodents might serve as a model for studying the alterations in the immune system with aging. PMID- 12078934 TI - Can I share my germs with you? PMID- 12078933 TI - Leap of faith. PMID- 12078935 TI - Private funding in clinical pharmacology. PMID- 12078936 TI - Predicting the risk of sporadic elevated bilirubin levels and diagnosing Gilbert's syndrome by genotyping UGT1A1*28 promoter polymorphism. AB - Elevated fluctuating levels of bilirubin are a common problem in clinical studies. Differentiation between a drug-related adverse event and the diagnostic symptom for Gilbert's syndrome (GS), an idiopathic unconjugated hyperbilirubinemia, is more or less impracticable since the diagnosis of GS is by exclusion. The aim of this investigation was to evaluate the correlation of unspecific elevated bilirubin levels and the occurrence of GS with a described polymorphism in the uridine diphosphat glucuronosyltransferase 1A1 (UGT1A1) in a predominately Caucasian population. 304 volunteers (152 male, 152 female) were genotyped for the UGT1A1 promoter polymorphism by PCR amplification and polyacrylamide gel electrophoresis. Serum bilirubin levels and liver enzymes were determined and GS was diagnosed using clinico-chemical criteria. 23/13 subjects displayed the homocygote variant, 73/66 the heterozygote variant and 56/72 wildtype (male/female, respectively). 23 male and 3 female volunteers fulfilled the clinical criteria for GS (15.1, respectively 2.0%). Men exhibited higher serum bilirubin levels than women with a mean (SD) of 14.37 (8.92) micromol/l compared to 10.17 (5.37) micromol/l, respectively (p < 0.001). The homocygote mutant promoter length correlated well with serum bilirubin levels and with the clinical diagnosis of GS (p < 0.001 each). Genotyping of the UGT1A1 promoter polymorphism is a cheap and unequivocal method for predicting elevated and fluctuating bilirubin levels. It is better suited to this purpose than the clinical diagnosis which is based on exclusion. The genotyping of UGT1A1 promoter polymorphism can help to improve safety and the reliable assessment of adverse events in clinical studies. Our data additionally support the demand to refine the bilirubin reference values. PMID- 12078938 TI - Hubble-bubble (water pipe) smoking: levels of nicotine and cotinine in plasma, saliva and urine. AB - OBJECTIVES: The purpose of the present study was to assess the levels of nicotine and cotinine in biological fluids (plasma, saliva, and urine) following hubble bubble (HB) smoking. METHODS: Fourteen healthy male volunteers, aged 28 +/- 8 years, body weight of 82.7 +/- 13.53 kg, participated in the study. All volunteers were habitual HB smokers for 3.29 +/- 1.90 years who smoked at least 3 runs per week with an average of 20 g Mua'sel per run. Volunteers were requested to avoid smoking, at least 84 hours prior to the time of the study. After baseline samples were taken, volunteers started smoking 20 g of Mua'sel for a period of 45 minutes. Heparinized blood samples (5 or 10 ml each) were drawn for nicotine and cotinine analysis before, during and after the smoking period. Saliva samples were collected just before smoking (time 0) and at the end of smoking (45 min). Urine also was collected at time 0 and 24-hour urine collection was also taken to measure nicotine and cotinine excretion. Nicotine and cotinine were extracted from samples and assayed by gas chromatography. All data are presented as mean +/- SEM throughout the text, Tables and Figures unless indicated otherwise. RESULTS: Plasma nicotine levels rose from 1.11 +/- 0.62 ng/ml at baseline to a maximum of 60.31 +/- 7.58 ng/ml (p < 0.001) at the end of smoking (45 min). Plasma cotinine levels increased from 0.79 +/- 0.79 ng/ml at baseline to its highest concentration of 51.95 +/- 13.58 ng/ml (p < 0.001) 3 hours following the end of smoking. Saliva nicotine levels significantly rose from 1.05 +/- 0.72 to 624.74 +/- 149.3 ng/ml and also saliva cotinine levels significantly increased from 0.79 +/- 0.79 ng/ml to 283.49 +/- 75.04 ng/ml. Mean amounts of nicotine and cotinine excreted in urine during the 24-hour urine collection following smoking were equal to 73.59 +/- 18.28 and 249 +/- 54.78 microg, respectively. CONCLUSION: Following a single run of HB smoking, plasma, saliva and urinary nicotine and cotinine concentration increased to high values. This observation suggests that HB may not be an innocent habit, as people believe. PMID- 12078937 TI - A post-marketing observational study to assess the safety of mibefradil in the community in England. AB - OBJECTIVES: To conduct a post-marketing observational cohort study to assess the safety of mibefradil in the community, using Prescription-Event Monitoring (PEM). METHOD: Data were collected and analyzed on patients prescribed mibefradil by 1,996 General Practitioners (GPs) throughout England. Incidence densities were calculated for all reported events and selected events were followed-up by means of further questionnaires. RESULTS: The study was terminated early due to the voluntary withdrawal of mibefradil from the market because of potential drug interactions. A cohort of 3,085 patients was recruited, with a mean age of 64.5 years. The major indication for use was hypertension (55% of the cohort), the indication was not specified in 33% of patients. 80% of GPs expressing an opinion rated mibefradil as effective. The major reason for stopping was withdrawal from the market (2,342 patients). The commonest reported adverse events and reasons for stopping were malaise/lassitude, dizziness, edema and headache. Seven clinically serious reports of bradycardia/collapse were considered to be possible adverse drug reactions (ADRs) to mibefradil. All were in the elderly (> 65 years), 6 were considered to be a result of possible drug interactions. In total, 11 possible drug interactions occurred. Nine (8 reports of bradycardia and 1 of syncope) involved beta-blockers. Another, a report of collapse and severe bradycardia, occurred in a patient who had started a dihydropyridine calcium channel blocker within 24 hours of stopping mibefradil and the other was a report of palpitations and dyspnea in a patient on concomitant digoxin and sotalol. None of the 53 deaths occurring during the study was attributed to mibefradil. CONCLUSION: Mibefradil was only available on the UK market for 6 months before it was withdrawn from the market because of potential drug interactions. With respect to the reasons leading to its withdrawal, in this cohort of 3,085 patients, 11 possible drug interactions were detected (6 clinically significant) involving beta-blockers, a dihydropyridine calcium channel blocker and digoxin and/or sotalol. PEM can contribute to the understanding of ADRs caused by drug interactions occurring in real-life settings. PMID- 12078939 TI - A comparison of non-tapering vs. tapering prednisolone in acute exacerbation of asthma involving use of the low-dose ACTH test. AB - OBJECTIVE: To determine if there is a difference in early relapse rates and adrenal suppression between patients receiving an 8-day course of 40 mg/day prednisolone and those receiving an 8-day tapering course of prednisolone. METHODS: This was a prospective, randomized, open clinical trial conducted in a tertiary care center. All asthmatic patients with exacerbation who were judged well enough for discharge home from the emergency department were eligible for participation. Patients with a history of chronic obstructive pulmonary disease, congestive heart failure, pneumonia, pneumothorax, or other pulmonary process and asthmatics already using inhaled or oral steroids within 2 weeks of admission to the emergency department were excluded. Patients on discharge were administered either on 8-day course of 40 mg/day prednisolone or an 8-day tapering course of prednisolone (tapering from 40 mg to 0 mg). Patients were asked to return on Day 12 for cosyntropin stimulated test and pulmonary function testing and on Day 21 for pulmonary function testing only. RESULTS: A group of 13 patients treated with non-tapering course (40 mg/day) of prednisolone for 8 days were compared to a group of 13 patients treated with a tapering course (40 mg taper by 5 mg/day) for 8 days. There were no differences in the FEV1 percent predicted (Days 12 and 21), the incidence of relapse, or the incidence of adrenal suppression between the 2 groups. CONCLUSION: In this small study, we found no significant difference in relapse rate or adrenal suppression between asthmatics receiving an 8-day tapering dose of prednisolone and those receiving 40 mg/day prednisolone upon discharge from the emergency department. PMID- 12078940 TI - Comparison of benazepril-amlodipine and captopril-thiazide combinations in the management of mild-to-moderate hypertension. AB - OBJECTIVE: To compare the efficacy and tolerability of benazepril 10 mg + amlodipine 5 mg combination (BZ+AM) versus captopril 50 mg + hydrochlorothiazide 25 mg (CP+HT) combination. MATERIAL: 405 outpatients with mild-to-moderate arterial hypertension not adequately controlled by a monotherapy with ACE inhibitors or calcium channel blockers or diuretics entered this multicenter, double-blind, randomized, parallel-group study. METHOD: After a 2-week placebo run-in, 397 patients with sitting diastolic (D) blood pressure (BP) > 95 mmHg and/or sitting systolic (S) BP > 160 mmHg were randomized to receive either BZ+AM (201 patients) or CP+HT (196 patients) once daily for 12 weeks. Main outcome measure was sitting DBP and SBP values at the end of active treatment. The response rate was defined as the proportion of patients with either a final sitting DBP < 90 mmHg or decreased by at least 10 mmHg or a sitting SBP < 150 mmHg or decreased by at least 20 mmHg from baseline. RESULTS: The DBP and SBP values obtained with BZ+AM were, respectively, 2.7 and 3.7 mmHg lower than those obtained with CP+HT (both p < 0.001 vs. CP+HT). The response rate in the BZ+AM group (94.8%) was better than that observed in the CP+HT group (86.0%, p = 0.004). The incidence of adverse events was similar with the 2 treatment regimens (17.9% for both). CONCLUSIONS: These data suggest a higher antihypertensive efficacy of the fixed combination BZ 10 mg+AM 5 mg as compared with CP 50 mg+HT 25 mg. PMID- 12078941 TI - Pharmacokinetic modelling of total and unbound plasma carboplatin--a population study in 75 children. AB - AIMS: A compartmental open model was developed to describe the relationship between plasma unbound (C.) and bound (CT) carboplatin concentrations. A population pharmacokinetic study was then undertaken to investigate the effect of demographic covariates on unbound and bound carboplatin clearance and volume parameters. METHODS: Carboplatin and demographic data were collected from 75 children (1-17 years old, 10 children with unilateral nephrectomy) treated using 1-hour daily infusions for various malignancies. Concentration-time data, C(U) and C(T), from children with rich data were used to develop the model. The data from all children were then simultaneously analyzed using a population approach. RESULTS: The average population values for total unbound carboplatin clearance, CL(U), and distribution volume of unbound carboplatin, VI, were 3.87 l/h and 6.26 l/h, respectively. The clearance of plasma-bound carboplatin was comparatively low, 0.11 l/h. CL(U) was dependent on weight, nephrectomy status and serum creatinine. A constant fraction of CL(U), 0.17 l/h, included the disappearance of unbound compound due to irreversible plasma binding. V1 was dependent on body weight. The unbound plasma carboplatin fraction (fu) was simulated and rapidly decreased with post-infusion time. CONCLUSIONS: The body weight was a better predictor for unbound carboplatin clearance than body surface area, and UNP and SCr caused a reduction in clearance of unbound carboplatin, as previously reported. The rate ofcarboplatin plasma binding was low and not dependent on demographic patient characteristics. The f(U) of plasma carboplatin could be predicted as a function of time, infusion rate and covariates affecting CL(U), weight, UNP and SCr. PMID- 12078942 TI - Hypertension in renal transplantation: donor and recipient risk factors. AB - AIMS: To determine the respective roles of donor and recipient factors in the subsequent development of hypertension after renal transplantation. PATIENTS AND METHODS: All the patients transplanted between January 1990 and December 1999 who still had a functioning graft 1 year post-transplant (n = 321) were retrospectively studied. Blood pressure was assessed at 1 year post-transplant. Hypertension was defined as a systolic BP > or equal 140 mmHg or diastolic BP > or equal 90 mmHg, or use of antihypertensive medication. Relevant donor and recipient characteristics were recorded. RESULTS: Two-hundred-and-sixty-three patients (82%) were hypertensive. In multivariate analysis, pretransplant hypertension (RR, 1.74, 95% CI, 1.07 to 2.87), anticalcineurin use (RR, 2.59, 95% CI, 1.13 to 5.92), urinary protein excretion (RR, 1.84, 95% CI, 1.06 to 3.18), BMI (RR, 1.08, 95% CI, 1.01 to 1.16), donor age (RR, 1.28,95% CI, 1.05 to 1.59, for each 10-year increase in donor age) and donor aortorenal atheroma (OR, 2.34; 95% CI, 1.24 to 4.46) were associated with hypertension. Among patients under calcineurin inhibitors, those receiving cyclosporine were more prone to have hypertension than those receiving tacrolimus (88.7% vs 78%; p = 0.04). CONCLUSION: Both recipient and donor factors contribute to hypertension in RTR. PMID- 12078943 TI - The LDH ratio as a marker for response to plasma exchange in HUS/TTP of the adult. AB - BACKGROUND: Plasma exchange improved the outcome of thrombotic thrombocytopenic purpura (TTP)/hemolytic uremic syndrome (HUS) of the adult markedly, but a high number of non-responders remain. Identifying these patients at an early stage would help to optimize therapy. AIM: determine the value of serologic measures in predicting the response to plasma exchange. MATERIAL AND METHODS: We performed a retrospective chart review of 30 patients with HUS/TTP of the adult treated with plasma exchange. According to the treatment protocol, a mean of 42 +/- 8.2 ml plasma per kilogram of body weight was exchanged daily for 3 days and continued every second day thereafter. Prior to each session, clinical status and serologic markers for hemolysis and kidney function were obtained. To assess the early individual response to plasma exchange, the decline of LDH from the first to the third cycle was calculated as: LDH concentration before the third session/LDH concentration before the first session (LDH ratio). RESULTS: During the observation period (median 195, range 6-1500 days), 80% of the patients responded to therapy with plasmapheresis. None of the serologic measures or clinical signs obtained before initiation of plasma exchange showed a significant correlation with the outcome. After 2 sessions of plasma exchange, only LDH and platelet level had improved markedly in responding patients. The LDH ratio was the best predictive marker for the individual response. An LDH ratio < 0.6 predicted a favorable outcome with a sensitivity of 0.96 and a specificity of 0.83. CONCLUSION: The LDH ratio might be a useful marker for separating patients responding to plasma exchange from those not responding at an early stage. PMID- 12078944 TI - Tolerance of mycophenolate mofetil in end-stage renal disease patients with ANCA associated vasculitis. AB - BACKGROUND: Mycophenolate mofetil (MMF) has been used successfully in patients with ANCA-associated vasculitis as maintenance therapy. Only transient and moderate side effects have been reported with a daily dose of 2 g. Since all the treated patients who have been reported so far had no or only moderate renal insufficiency when MMF was initiated, no data are available regarding side effects in patients with end-stage renal disease (ESRD). METHODS: Five ESRD patients with ANCA-associated vasculitis and a relapsing course of their disease were treated. All patients had pretreatment with cyclophosphamide for at least 17 months. MMF was initiated as a remission maintenance therapy, and started with a dose of 1 g/d. The aim was to increase the MMF dose to 2 g/d. Blood counts, liver enzymes and gastrointestinal side effects were monitored. RESULTS: Four patients developed severe anemia, 2 requiring blood transfusion with permanent or temporary cessation of MMF treatment. One patient developed leukopenia. Gastrointestinal symptoms led to a dose reduction to 1 g/d (n = 2) or cessation of treatment (n = 1). Three patients remained on longer MMF treatment; however, their daily dose did not exceed 1 g. CONCLUSION: MMF, a promising drug regarding maintenance therapy in ANCA-associated vasculitis, seems to have more side effects in ESRD patients, leading to dose reduction or even cessation of treatment. Therefore, in this patient group a lower dose and closer monitoring for side effects seems to be required compared to patients with no or moderate renal insufficiency. PMID- 12078945 TI - The effects of prolonged physical exercise on renal function, electrolyte balance and muscle cell breakdown. AB - BACKGROUND: Postexercise proteinuria, hematuria and changes in serum electrolyte balance as well as increased levels of plasma indicators for muscle leakage are believed to be transient and of benign character. METHODS: A group of 51 healthy athletes took part in a 100 km race over 14.25 hours. All of them had to reach the finish together. Urine and blood samples were collected before (a) and immediately after running (b) as well as 6 hours after the race (c). RESULTS: The serum concentrations of potassium (4.8 +/- 0.5 (a) vs. 4.0 +/- 0.3 (c) mmol/l), protein (73.1 +/- 5.2 (a) vs. 71.1 +/- 3.9 (c) g/l) and albumin (44.0 +/- 2.85 (a) vs. 42.9 +/- 2.8 (c) g/l) decreased significantly (p < 0.0001, p < 0.05, p < 0.05, respectively) but remained within physiological ranges. The serum sodium concentration decreased immediately after the race (136.9 +/- 4.5 (a) vs. 131.1 +/- 2.4 (b) micromol/l, p < 0.0001). The fractional sodium excretion decreased 6 hours, but not immediately after the race (0.78 +/- 0.59 (a) vs. 0.48 +/- 0.82 (c), p < 0.05). Myoglobin (31.8 +/- 6.9 (a), 291.5 +/- 197.2 (b) and 182.2 +/- 135.3 (c) microg/l, p < 0.0001) and creatine kinase (1.13 +/- 0.45 (a), 10.76 +/- 6.9 (b) and 9.46 +/- 15.5 (c) pmol/l, p < 0.0001) increased dramatically. Troponin I was also significantly increased at finish (0.0186 +/- 0.0121 (a) vs. 0.0213 +/- 0.0165 (b) ng/ml, p < 0.05) and positively correlated with myoglobin and creatine kinase, but remained far below the pathologic range. Serum creatinine and urea remained almost unchanged. Glucosuria and hematuria occurred 6 hours after the run in 9.1% and 6.8%, respectively. The erythrocytes examined by phase-contrast microscopy were not damaged in terms of dysmorphic cells. Glomerular-type proteinuria was found in 11.4% of the participants 6 hours after the race. CONCLUSIONS: We conclude that long lasting, mild exertion is harmless for renal function, electrolyte balance and skeletal muscle as well as myocardial metabolism in healthy persons. PMID- 12078946 TI - Perinatal rupture of the uropoietic system. AB - AIMS: Ruptures of the uropoietic system resulting in either urinary ascites or urinoma are rare complications in the neonate. Although ruptures without clear predisposing factors are described, in most cases they are associated with obstructive uropathy. The diagnosis is often delayed and the prognosis is related to the degree of renal damage. There is discussion about possible protective mechanisms of the rupture for renal function in patients with obstructive uropathy. METHODS: We retrospectively analyzed the clinical presentation, predisposing factors and the renal function before and after treatment of 10 neonates with a rupture of the pyelum or urinary bladder in our hospital. RESULTS: The group consisted of 9 boys and 1 girl. The average birth weight was 3,880 g. The patients presented with distended abdomen (n = 10), abdominal mass (n = 2), ascites (n = 5), oligohydramnion (n = 2), hypertension (n = 1) and anuria (n = 1). Underlying diagnosis included obstruction of the ureteropelvic junction (UPJ obstruction) in 3 children and posterior urethral valves in 7 children. Five children presented with urinoma, 3 children had a urinoma in combination with ascites and 2 children had isolated ascites. All children had reduced renal function at the time of diagnosis. In all 10 cases, the serum creatinine decreased after treatment. Scintigraphic investigation with mercapto acetyltriglycerine (MAG III) demonstrated diminished function and perfusion of all 3 kidneys with UPJ obstruction and isolated urinoma even after treatment. Children with posterior urethral valves and urinoma revealed better function of the ruptured kidney and diminished function of the kidney which was not ruptured. One child with a rupture of the urinary bladder and urinary ascites showed good function and perfusion of both kidneys. CONCLUSION: Perinatal ruptures of the uropoietic system are rare. The clinical presentation is aspecific. One should consider a rupture of the urinary bladder or pyelum in a neonate with a distended abdomen, hydronephrosis and ascites. The long-term prognosis depends on the underlying diagnosis and the location of the rupture. Probably, a UPJ obstruction with an isolated urinoma is associated with irreversible renal damage of the ruptured kidney. A rupture resulting in urinary ascites apparently provides better decompression with better function of the ruptured kidney. Scintigraphic investigation is necessary for a separate evaluation of the single kidney function. PMID- 12078947 TI - The emergency department care of hemodialysis patients. AB - AIMS: To describe the emergency department (ED) presentation, evaluation and disposition of maintenance hemodialysis (HD) patients. MATERIALS AND METHODS: A retrospective review of adult HD patients seen 1/1-12/31/97. The following was collected: demographics, mode of arrival, chief complaint, etiology of renal failure, evaluation, treatment, disposition, length of stay and facility charges. During the study period, this tertiary care ED had an annual adult census of 45,000. No clinical pathways were in place. RESULTS: 143 patients made 355 visits: 351 charts were available. Mean patient age was 51 (range 20-86), 62% were male, 51% were white. 70% presented from home, 26% from dialysis. EMS transported 32%. Medicare insured 78%. Etiologies of renal failure included hypertension (33%), diabetes (27%), HIV (7%) and glomerulonephritis (8%). Complaints were related to infection (18%), dyspnea (17%), vascular access (16%). chest pain or dysrhythmia (15%) and gastrointestinal complaints (12%). ED evaluation included CBC (79%), electrolytes (75%), CXR (57%) and EKG (48%). Antibiotics were administered to 21%. HD was performed earlier than scheduled in 14%. Two hundred and eighteen patients (62%) were admitted (ICU 11%, telemetry 22%), 19 (5%) refused admission and 2 expired in the ED. The average hospital length of stay was 7.8 days (range 1-59), with 29% hospitalized more than 1 week, compared to 6.54 days for non-HD patients. The mean facility charge for admitted subjects was $14,758, while the average cost for non-HD admissions was $7,152. Of the 133 patients (38%) who were discharged directly from the ED, the mean length stay was 223 minutes (range 30 to 750) and the mean charge was $658. The mean length of stay for non-HD patients was 124 minutes. CONCLUSION: The ED evaluation of adult HD patients involves multiple diagnostic modalities, and patients are usually admitted. The admit rate, ED length of stay for discharged patients and hospital charges for care were substantially higher in the HD patients than in the general population. Further research in the ED care of these complex patients should be undertaken. PMID- 12078948 TI - The mature form of adrenomedullin correlates with brain natriuretic peptide in plasma of chronic hemodialysis patients. AB - AIM: Adrenomedullin (AM), a hypotensive and natriuretic peptide, consists of an amidated mature form (mAM) and an intermediate form in human plasma, of which only mAM exerts biological activity. Like atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP), plasma levels of mAM are reported to be significantly elevated in hemodialysis (HD) patients, suggesting that mAM may be stimulated partly by increased body fluid volume in a manner similar to the natriuretic peptides. Here, we examined the relationship between mAM levels and ANP or BNP levels and the effect of HD on plasma mAM in HD patients. PATIENTS AND METHODS: We measured plasma levels of mAM, total AM (tAM), ANP and BNP before and after HD in patients on long-term HD (n = 22, mean age 56.3 +/- 3.2 years) using radioimmunoassay. RESULTS: Baseline mAM (2.7 +/- 0.3 fmol/ml) and tAM (23.6 +/- 2.0 fmol/ml) were significantly higher in HD patients than in healthy subjects (1.1 +/- 0.2 fmol/ml, 9.0 +/- 2.1 fmol/ml, respectively). HD significantly reduced the levels to 1.2 +/- 0.2 fmol/ml and 13.8 +/- 1.4 fmol/ml, respectively, although tAM levels were still elevated compared to healthy subjects. Similar plasma ANP and BNP levels were obtained in HD patients. There were significant correlations between mAM and tAM levels before and after HD and between HD induced changes in mAM and tAM levels. In the pre-HD state, levels of both mAM and tAM correlated significantly with BNP levels, but the correlation of BNP with mAM was closer than that with tAM. In contrast, no correlations were observed between the 2 forms of AM and ANP. Changes in mAM levels during HD also correlated significantly with BNP but not ANP levels, although the changes in tAM did not correlate with those of the 2 natriuretic peptides. CONCLUSION: Our results suggest that the secretion/metabolism of mAM may be regulated in a manner similar to that of BNP in HD patients. PMID- 12078949 TI - Serial ferritin concentrations in hemodialysis patients receiving intravenous iron. AB - BACKGROUND: Treatment of the anemia of chronic renal failure with intravenous iron and erythropoietin is highly effective, but frequently leads to ferritin levels which are much higher than those seen in the general population. High ferritin concentrations raise concern about the potential toxicity of increased body iron stores. PATIENTS AND METHODS: We retrospectively evaluated parameters of iron metabolism over a 4-year period among all our chronic hemodialysis patients who had been receiving intravenous iron and erythropoietin. Initially, patients received intermittent infusions of 300 mg intravenous iron x 3 doses for a low ferritin or low percent saturation of total iron binding capacity (TIBC), but this protocol was subsequently changed to weekly or biweekly infusions of 50 100 mg. RESULTS: We observed an improvement in average hemoglobin values, modest increases in serum iron and saturation of iron binding capacity, and a 125% increase in ferritin levels over 4 years. TIBC decreased. Overall, ferritin values increased 79 microg/l for each 1% increase in TIBC saturation. Ten patients with ferritin concentration greater than 1,000 pg/l received a three month course of vitamin C with no decline in the ferritin concentration. CONCLUSION: Current protocols for iron delivery may result in progressive increases in ferritin levels. Concern about the risks of iron overload should temper the quantity of iron used in dialysis programs. PMID- 12078950 TI - Intravenous iron administration does not significantly increase the risk of bacteremia in chronic hemodialysis patients. AB - BACKGROUND: Correction of iron deficiency is critical in chronic hemodialysis patients, and intravenous administration is superior to the oral route in this goal. Recently, concern was raised that intravenous iron administration might promote infection in dialysis patients. METHODS: We reviewed the data from a recent prospective study of 985 patients in which no link between iron therapy and bacteremia had been found. We tested the potential role of the administration route of the iron (intravenous vs. oral), the weekly amount of iron administered and the administration rate on the risk for bacteremia in these patients. RESULTS: were 4-fold: in multivariate analysis, neither intravenous iron administration in the whole population nor the weekly amount of iron in the subgroup of i.v. iron-treated patients were significant risk factors for bacteremia; iron was not given more frequently intravenously in bacteremic than in non-bacteremic patients; among patients treated with intravenous iron, the frequency and the amount of iron administered were significantly higher in those who developed bacteremia than in those who did not; and in patients receiving i.v. iron, there was an increased risk of bacteremia associated with concurrent administration of erythropoietin, which was not observed in patients receiving iron orally. CONCLUSION: This study failed to demonstrate a significant association between intravenous iron administration and the risk of bacteremia in dialysis patients. However, there might be a slightly increased risk of bacteremia in patients given high-frequency, high-dose intravenous iron. PMID- 12078951 TI - Simultaneous non-surgical removal and insertion of peritoneal dialysis catheters for defective drainage: a bedside, day case technique. AB - BACKGROUND: Outflow failure of peritoneal dialysis catheters is a commonly encountered problem. It may be possible to reposition the catheter by a variety of means, but this can be problematical and has variable long-term success. Commonly surgical catheter exchange is utilized, entailing inconvenience, expense and often, a reliance on temporary hemodialysis. We describe a technique allowing exchange of poorly functioning catheters with a relatively simple outpatient/day case percutaneous technique, allowing the continuation of peritoneal dialysis. METHODS: We report percutaneous exchange of 25 peritoneal dialysis catheters in 21 patients. The exchanges were performed under local anesthesia with a degree of sedation (if required). It involved the dissection down the distal cuff of the catheter and mobilization of the catheter below it. This was followed by division of the catheter, allowing passage of a guide wire into the peritoneal cavity and insertion of a further peel away sheath and insertion of a new catheter. The new catheter was tunneled out of the existing exit site after removal of the extraperitoneal portion of the old catheter by traction. RESULTS: Outflow failure was associated with fecal loading and malposition of the catheter in 14 out of the 21 patients. Exchange of catheter was successful in all the patients with good pelvic positioning of the replacement catheter in all but 1 of the cases. The mean period until the reinstitution of peritoneal dialysis was 5.1 days (range 0-14 days). Temporary hemodialysis was not required for any of the patients. One patient exhibited a small leak of peritoneal dialysis fluid after insertion, but this had spontaneously resolved within 6 days. Protracted satisfactory function of the peritoneal dialysis catheters was obtained in all but 1 of the patients (mean follow-up 51 weeks, range 11-73 weeks). CONCLUSIONS: We conclude that exchange of peritoneal dialysis catheters for problems with dialysate drainage, utilizing a non-invasive percutaneous technique is both effective and safe. PMID- 12078952 TI - Cerebral, myocardial and cutaneous ischemic necrosis associated with calcific emboli from aortic and mitral valve calcification in a patient with end-stage renal disease. AB - We report the case of a 57-year-old diabetic male with chronic renal failure who developed secondary hyperparathyroidism and calcification of mitral and aortic valves and interatrial septum. Multiple ischemic lesions developed in the skin of hands, feet and penis, and in the brain, and these were presumed to be due to septic emboli from cardiac valvular infective endocarditis. Multiple blood cultures were negative, however, and despite antibiotic therapy the patient expired. Autopsy (limited to trunk) demonstrated multiple calcific emboli in the heart and spleen, apparently derived from the prominent calcific deformities in the aortic and mitral valves. These were associated with acute and organizing myocardial infarcts and acute splenic infarcts, suggesting that the multiple ischemic lesions in the brain were also due to calcific emboli. A possible contributory component of infective endocarditis, however, was indicated by postmortem cultures of aortic and mitral valves positive for Enterococcus faecium. Calcific embolism is a rarely recognized but potentially lethal complication of end-stage renal disease, and the clinical diagnosis and the preventive therapeutic options for the control of the product of calcium and phosphate and/or parathyroidectomy should be considered. PMID- 12078953 TI - Systemic amyloidosis involving the diaphragm and acute massive hydrothorax during peritoneal dialysis. AB - Hydrothorax secondary to trans-diaphragmatic fluid leakage through a peritoneo pleural communication is an occasional, potentially serious complication of peritoneal dialysis. The etiology of this condition is not clear, being thought to be due either to congenital or acquired diaphragmatic fenestrations or acquired scarcity of muscle fibers in the tendinous part of the diaphragm which are compounded by increased intra-abdominal pressure during the dwell period of peritoneal dialysis. We report a 54-year-old woman who developed irreversible acute renal failure from adjuvant chemotherapy for ovarian cancer previously resected surgically. Three days after the onset of continuous ambulatory peritoneal dialysis, she developed acute respiratory distress associated with a massive right hydrothorax secondary to a peritoneo-pleural communication demonstrated by scintigraphy. At autopsy 2 weeks later, systemic amyloidosis was surprisingly found and histologic examination of the right hemidiaphragm showed the presence of amyloid, among sparse muscle fibers. This is the first case report of a distinct pathological process, i.e. amyloidosis, involving the diaphragm associated with a peritoneo-pleural communication causing massive hydrothorax at the onset of peritoneal dialysis. PMID- 12078954 TI - A case of mixed membranous nephropathy and purpura nephritis. AB - We report the case of a 71-year-old man with mixed glomerular lesions, membranous and necrotizing changes. The patient had abdominal pain and purpurat on the extremities and trunk, followed by melena, and after admission to hospital, proteinuria and occult blood were noted. Laboratory findings were negative for autoimmune disease and viral hepatitis. Renal biopsy showed segmental necrotizing changes and mesangial proliferation with spike formation. Immunofluorescence revealed a granular deposition of IgA predominantly in the mesangial area in contrast to the granular IgG deposition along the glomerular capillary loops. Moreover, electron-microscopically, mesangial as well as subepithelial electron dense deposits were observed. These data suggest that the patient had 2 distinct types of glomerulonephritis simultaneously: idiopathic membranous nephropathy and purpura nephritis. PMID- 12078955 TI - High-flux hemodialysis--an effective alternative to hemoperfusion in the treatment of carbamazepine intoxication. AB - Carbamazepine intoxication is associated with seizures, coma, arrhythmias and death. In acute intoxication, charcoal hemoperfusion is employed for removal of the drug. This can be associated with thrombocytopenia, coagulopathy, hypothermia and hypocalcemia. Alternatively, we used high-efficiency hemodialysis with a batch dialysis system (Genius), lowering not only serum levels of carbamazepine but removing a considerable amount of the drug as measured in the dialysate. This treatment regimen was compared to treatment by hemoperfusion. A 3.5-hour high flux hemodialysis was as effective as a 2-hour hemoperfusion. We conclude that high-efficiency hemodialysis is a safe and effective alternative for treating life-threatening carbamazepine intoxication. PMID- 12078956 TI - Improvement of anemia in hemodialysis patients after pulse oral 1-alpha-D3 treatment. PMID- 12078957 TI - HIV and hepatitis B surveillance in First Nations alcohol and drug treatment centres in British Columbia, Canada, 1992-2000. AB - We provide ongoing HIV and Hepatitis B surveillance in residential First Nations alcohol and drug treatment centres in British Columbia, Canada. All clients entering the centres are offered confidential viral testing as part of an education program on sexually transmitted diseases. Participation is voluntary and approximately two thirds of clients choose to be tested. Information about risk factors for communicable disease and immunization status is not recorded. The testing program began in January 1992. As of September 2000, 2,345 people have been tested for HIV. Nine tested positive, giving a prevalence of 3.8 per 1,000 (95% confidence interval: 1.3 to 6.3 per 1,000), lower than among all British Columbians who choose to be tested (8.5 per 1,000). Also, 2,166 people were tested for hepatitis B surface antibody, 23% of these were positive, 10% were positive for hepatitis B core antibody (indicating prior infection with hepatitis B). Seven clients (3.2 per 1,000; 95% Cl: 0.8 to 5.6 per 1,000) were positive for hepatitis B surface antigen and are therefore presumed to be chronically infected. The prevalence of hepatitis B markers was intermediate between what has previously been found in high risk groups and that found in the general population. PMID- 12078958 TI - Violence, sexual abuse and health in Greenland. AB - The purposes of the study were to analyse the lifetime prevalence of violence and sexual abuse among the Inuit in Greenland and to study the associations between health and having been the victim of violence or sexual abuse. Associations were studied with specific attention to possible differences between women and men. Further, response rates were analysed specifically in order to understand consequences of including questions on violence and sexual abuse in the questionnaire survey. The analyses were based on material from a cross-sectional health interview survey conducted during 1993-94 with participation from a random sample of the Inuit population in Greenland (N = 1393). The prevalence of ever having been a victim of violence was 47% among women and 48% among men. Women had more often than men been sexually abused (25% and 6%) (p < 0,001) and had more often been sexually abused in childhood (8% and 3%) (p = 0.001). Having been the victim of violence or sexual abuse was significantly associated with a number of health problems: chronic disease, recent illness, poor self-rated health, and mental health problems. The associations between having been the victim of violence or sexual abuse and health was stronger for women than for men. It is possible to secure a reasonably high response rate in a general health survey that includes questions on violence and sexual abuse. PMID- 12078959 TI - Neurotic psychopathology and alexithymia among winter swimmers and controls--a prospective study. AB - Random samples of 25 voluntary Finnish winter swimmers (7 males, 18 females) and 11 controls (3 males, 8 females were followed prospectively during the winter season from October 1999 to May 2000 to (determine whether winter swimming is beneficial for mental well-being, as many of its practitioners claim. The Crown Crisp Experimental Index (CCEI) was used for measuring free-floating anxiety, phobic anxiety, obsessionality, depression, somatic anxiety and hysteria, and the 20-item version of the Toronto Alexithymia Scale (TAS-20) for measuring alexithymia. Self-reported somatic and mental health and the reasons for and the frequency of winter-swimming were asked, too. As resealed by open questions, the winter swimmers reported positive effects of winter swimming. Several of the swimmers also told that they had started winter swimming to improve their physical and mental health. Their experience was that the swimming had relieved physical symptoms and made their mood more positive. However, we found no major differences between winter swimmers and controls in any CCEI or TAS variables. The structured questionnaires do not necessarily, however, reach subjective feelings and experiences. PMID- 12078960 TI - Regional and seasonal variation in the length of hospital stay for chronic obstructive pulmonary disease in Finland. AB - Notable regional and seasonal variation has been reported in the rate of hospital admissions for chronic obstructive pulmonary disease (COPE). The aim of this study was to assess the variation in the length of hospital stay for COPD in Finland on the north south axis and by season. For this purpose, the patient records of subjects aged over 45 hospitalised altogether 153,401 times with COPD as their primary diagnosis during 1987 - 1998 were retrieved from the Finnish Hospital Discharge Register maintained by the National Research and Development Centre for Welfare and Health. During this period, the average length of hospital episodes was 9.8 (SD) 47.8) days in northern Finland and 11.9 (SD 54.5) days in southern Finland (p = 0.001). Throughout Finland, the mean duration of hospital stay was longest in the winter, 12.3 (SD) 62.3) days, and shortest in the summer, 11.0 (SD) 42.3) days (p = 0.001). The mean duration of hospital stay in northern Finland was also longest in the winter, 10.6 (SD 56.7) days, and shortest in the summer, 8.8. (SD) 26.7) days (p = 0.015). Hospital episodes for COPD vary in duration in Finland, probably mainly due to regional differences in health care resources and treatment routines. The light and warm northern summer may also speculatively serve to shorten hospital episodes. PMID- 12078961 TI - Project for developing a cold risk assessment and management strategy for workplaces in the Barents region. PMID- 12078963 TI - Neuromuscular performance limitations in cold. AB - This review will focus on the effects of cooling on muscular performance and its variables, functional properties of the muscles and some neural aspects of muscle function. The changes are described in terms of different cold exposures with varying intensity, therefore also looking at the dose dependent relationship between cooling and performance decrement. In addition, relationship between rewarming exercise and performance enhancement is described. Future research needs are addressed. PMID- 12078962 TI - Usability of isothermal standards for cold risk assessment in the workplace. AB - In this study the usability of ISO thermal standards suitable for cold risk assessment in the workplace was evaluated. Persons who were not earlier familiar with the methods were trained to be able to perform the assessments. The assessments were conducted at two different stages according to method complexity, duration and need for expertise. The methods tested at stage 1 included observation and simple measurements (ISO CD 15265, ISO 8996, ISOTR 11079) and were assumed to be relatively simple to use. The methods tested at stage 2 included measurements and quantitative analyses (ISO 7726, ISO 8996, ISO 10551, ISO 9920, ISO 9886, ISO TR 11079). Training to use the methods was provided to the observers and measur ers. The assessments at stage 1 were performed by 5 observers repeated on three different occasions with 5 different workers. The measurements (stage 2) were conducted by four persons and repeated four times. A questionnaire concerning the usability of the method tested was filled in after each assessment. The results show that the methods of stage 1 were considered easy to adopt, the duration of the assessment was short (average 30 min), the results were easy to analyse and the amount of training was considered to be adequate. The methods of stage 2 were considered more laborious to perform and the total duration of the assessment was on average 4 h 40 min. Furthermore, the training provided was not considered to be entirely sufficient and additional instructions were frequently asked for, especially when analysing the results. Usable elements were recognised from the ISO thermal standards, which could be used at the more advanced stages of cold risk assessment. PMID- 12078964 TI - Gross and fine neuromuscular performance at cold shivering. PMID- 12078966 TI - The Arctic Council and health issues. PMID- 12078965 TI - Cold exposure and musculoskeletal disorders and diseases. A review. AB - The aim of this study was to investigate how cold exposure may be associated with musculoskeletal problems either on symptomatic or disease level based on available relevant scientific literature. The data collection resulted in ten scientific articles, from which nine were epidemiological and one a case report consisting of three cases. The results indicated that musculoskeletal symptoms are more frequent in cold store work and in related conditions than in normal temperature work and symptoms seems to be increased when the working time in cold environment increases. In cold store work low back pain and knee pain are more frequent problems than in normal temperature working environment. The association between cold exposure and shoulder pain is still unclear and has been poorly studied. Meteorological conditions, principally low ambient temperature may have an effect of reported low back pain in a remarkable part of population. In the etiology of tenosynovitis cold exposure has a specific role either as a causative or a contributing factor. The risk for carpal tunnel disease is 2.2-fold in repetitive wrist movements compared with no repetition and 9.4-fold when cold exposure added. Cold exposure in work seems to be is associated with a little increased risk (Odds ratio 2.2) for degenerative discopathies of lower back in men. However, this risk is small and seems to be associated with other physical factors in the background. In conclusion, the associations between cold exposure and musculoskeletal complaints or diseases are shown in some studies and the need of further research is apparent. PMID- 12078967 TI - A vital statistics system for determining births and mortality in the First Nations population of British Columbia, Canada. AB - We describe a unique method for producing province-wide and selected regional birth and death statistics for the First Nations population, We identified births and deaths of persons with Indian Status in the province of British Columbia, Canada, using three databases: the Vital Statistics Agency's database of births and deaths, the Department of Indian Affairs' Indian Status Verification File and the provincial health insurance plan's Status Indian Entitlement file. A birth or death was considered Status Indian if the person was so identified in any of the three sources. From 1991 through 1998, 24,159 live births were identified as Status Indian: 54% by all three sources, 29% by two sources and 17% by one source. In the same period, 5,680 Status Indian deaths were identified: 28% by all three sources, 38% by two sources and 34% by one source. Deaths were grouped by Underlying Cause and rates in each category were age-standardized for comparison to the general population. This project underscores the importance of using more than one database to ensure complete counting. Otherwise, birth and death rates will be underestimated. This has implications for national reporting if each province does not have a comparable system. PMID- 12078968 TI - Human T-lymphotropic virus type I and II infections in First Nations alcohol and drug treatment centres in British Columbia, Canada, 1992-2000. AB - Since 1992, prevalence data on HTLV-I and II have been collected as part of an ongoing viral seroprevalence study in clients of six First Nations alcohol and drug treatment centres in British Columbia, Canada. Prior studies indicate that the lifetime risk of clinical disease (neurologic or hematologic) resulting from HTLV-I infection is low (less than 5%) and HTLV-II to date has not been clearly associated with clinical disease. In 1993, the first cases of HTLV-I-associated myelopathy or tropical spastic paraparesis (HAM/TSP) were reported in four Aboriginal residents of British Columbia; these were the first reports of HTLV-I linked disease among Aboriginal persons in Canada. All clients of the treatment centres involved in this study were offered confidential, voluntary testing following pre-test counseling, and the results are given to participants before the residential session is complete. 1953 men and women were tested; 11 were positive for HTLV-1 (0.56%) and 33 were positive for HTLV-2 (1.8%). PMID- 12078969 TI - Benign mucinous cystadenoma with stromal luteinization during pregnancy: a hormonally responsive tumor and a rare cause of fetal intrauterine growth restriction. AB - A 32-year-old woman, gravida 4, para 3, had a benign right ovarian mucinous cystadenoma. It was diagnosed 1 month before conception, measured 9 x 7cm, and reached 33 x 24 x 20cm at 38 weeks gestation. Cesarean delivery of a 2250 g normal female infant took place, and the removal of a 6300 g right ovarian cystic mass was also performed. Microscopic and immunohistochemical studies of the cyst wall revealed stromal luteinization and a strong reaction with antiserum to testosterone, estrogen and progesterone. PMID- 12078970 TI - Pregnancy complicated with lactate dehydrogenase M-subunit deficiency: the first case report. AB - A woman with lactate dehydrogenase M-subunit deficiency underwent two cesarean sections because of the risk of dystocia due to decreased adenosine triphosphate production in anaerobic glycolysis including uterine muscles. Frequent pains with increased serum pyruvate levels were observed during the third trimester of her pregnancies. PMID- 12078971 TI - Alcoholization: the choice of intrauterine treatment for chorioangioma. AB - Chorioangioma is a vascular tumor of the placenta. Most are small and asymptomatic, whereas the large tumors are clinically significant and often associated with polyhydramnios and fetal heart failure. To prevent fetal loss from these complications, many interventions have been proposed, including intrauterine transfusion in anemic cases and fetoscopic surgery to ablate the feeding vessels. The case presented herein had large chorioangiomas, 8 and 4cm in diameter, associated with polyhydramnios and early signs of hydrops fetalis, diagnosed at 27 weeks gestation. After extensive counseling, we performed alcohol ablation of the feeding vessel of the larger tumor. Signs of fetal heart failure and hydrops fetalis disappeared dramatically. The pregnancy was extended for 2 weeks, followed by premature rupture of the membranes and spontaneous labor at 32 weeks gestation and a surviving female baby, weighing 1360g, was delivered uneventfully. This preliminary experience suggests that alcoholization may be one of the best choices for this condition due to its high efficacy, simplicity, safety and very low cost. To our knowledge, this is the first report using alcoholization for the treatment of hydrops fetalis secondary to chorioangioma. PMID- 12078972 TI - Intrauterine treatment for an acardiac twin with alcohol injection into the umbilical artery. AB - An acardiac twin is a unique complication of monochorionic twinning, in which a normal pump twin perfuses the acardiac twin. The mortality rate of the pump twin is greater than 50%. Herein we present the successful treatment of an acardiac twin with alcoholization as follows. An acardiac twin was diagnosed at 24 weeks gestation. Circulation interruption of the acardiac twin was successfully achieved by injection of absolute alcohol (5 mL) into the intra-abdominal umbilical artery. Serial ultrasound after the procedure revealed normal growth of the pump twin, whereas the acardiac twin was shrinking. Spontaneous premature delivery at 34 weeks gestation resulted in a normal surviving female baby weighing 2410g and an acardiac female fetus weighing 300g. This experience suggests that this simple technique may be an alternative treatment for an acardiac twin. PMID- 12078973 TI - Role of new sono-imaging technique 'sonocolpography' in the diagnosis and treatment of the complete transverse vaginal septum and other allied conditions. AB - OBJECTIVE: To evaluate the value of a newly suggested ultrasonographic technique 'Sonocolpography' in diagnosing the complete transverse vaginal septum and other allied conditions and planning their treatment. SUBJECTS AND METHODS: Ten cases of complete vaginal septum, eight cases of atresia and four cases of vaginal aplasia seen at Kasr El Aini School of Medicine, Cairo University were selected for this study. Sonocolpography is done using a vaginal balloon and transabdominal ultrasonography. The efficacy of the latter is assessed in comparison to the conventional transabdominal technique. The different sonographic criteria in the conventional technique and in sonocolpography were determined for the studied cases. A cross-over test was also suggested for anticipating the needed surgery. This is done by pushing the balloon maximally inward and determining the ability of the echoes of the vaginal pouch to accommodate the balloon to stretch and to cross over the defect, in turn determining the needed surgery. RESULTS: Improved ultrasonographic images were successfully obtained for all cases studied. The diagnostic accuracy for these cases was 95.5% for septa, 86.4% for atresia and 90.9% for aplasia. The corresponding figures for the conventional techniques were 54.5, 68.2 and 77.2%, respectively. Sonocolpography was more sensitive and more specific for all cases studied compared to the conventional technique. Planning of the necessary surgery was successfully determined in 16 cases having positive cross-over tests, where excision and direct anastomosis were performed. Surgery planning was also successful in two cases having negative tests and four cases of invalid tests, where more extensive surgery was needed. CONCLUSION: Sonocolpography is a new achievement that is essential for imaging cases of the complete vaginal septum and other allied conditions, and also in planning the associated treatment. PMID- 12078974 TI - Mucocolpos due to complete transverse septum in middle third of vagina in a 17 year-old girl. AB - A 17-year-old girl had a large abdominopelvic mass (12 x 8cm) extending from vagina, due to the presence of a complete transverse vaginal septum in its middle third which on resection facilitated the drainage and disappearance of the mass finally diagnosed as a case of mucocolpos. PMID- 12078975 TI - Value of humerus length shortening for prenatal detection of Down syndrome in a Thai population. AB - OBJECTIVE: To assess the value of humerus length shortening for prenatal detection of Down syndrome in a Thai population. METHODS: A prospective study was performed on 3053 women undergoing second-trimester amniocentesis, between 16 and 24 weeks gestation, for the indications of advanced maternal age and a past history of chromosomal abnormality. Biparietal diameter (BPD) and humerus length measurements were obtained before the procedures. Regression equations relating BPD to humerus length were used to calculate observed humerus length/expected humerus length ratio in chromosomally normal and Down syndrome fetuses. Sensitivity, specificity, false-positive rate and likelihood ratio of a positive test result at various observed humerus length/expected humerus length ratios for detection of Down syndrome were calculated. A receiver-operator characteristic curve was used to determine the threshold screening ratio. RESULTS: There were 3003 chromosomally normal pregnancies and 24 fetuses with Down syndrome. The relationship between humerus length and BPD was: expected humerus length = 0.7403BPD - 5.1057, R2= 0.77, P < 0.001. Humerus length in Down syndrome fetuses was significantly shorter than in normal fetuses (P < 0.001). A ratio of 0.91 for observed humerus length/expected humerus length yielded a sensitivity of 41.7%, specificity of 88.3%, a false-positive rate of 11.7% and likelihood ratio of a positive test result of 3.63 (95% confidence interval 2.24-5.88) for detection of Down syndrome. CONCLUSIONS: Humerus length shortening in the second trimester appears to be a useful adjunctive screening parameter for fetal Down syndrome in a Thai population. PMID- 12078976 TI - Detection of residual disease by cytology in patients with cervical intraepithelial neoplasia III post-large loop excision of the transformation zone. AB - OBJECTIVE: To evaluate the diagnostic accuracy of cytology in detecting residual disease in patients with cervical intraepithelial neoplasia (CIN) III post-large loop excision of the transformation zone (LLETZ). METHODS: This prospective study was performed between February 1994 and August 1999 at the Department of Obstetrics and Gynecology, Ramathibodi Hospital, and involved 90 patients who underwent LLETZ and had histologic confirmation of CIN III. Simple hysterectomy was performed in all patients 2-3 months after LLETZ. Two Papanicolaou smears were taken using Ayre spatula on each patient 6 weeks after LLETZ and 1 day before hysterectomy. Histologic findings of hysterectomy specimens were used as the 'gold standard'. RESULTS: Forty-six cases (51.1%) were found to have residual diseases: seven cases with CIN I, 11 cases with CIN II, 27 cases with CIN III and one case with invasive squamous cell carcinoma. Using the most severe diagnosis of two Papanicolaou smears performed on each patient as the cytologic diagnosis, the sensitivity, specificity, positive predictive value and negative predictive value were 28.3, 93.2, 81.3 and 55.4%, respectively. The accuracy rate was 60.0%. Residual diseases were found in 62.5 and 39.0% of cases with the presence and absence of CIN at the margin of the LLETZ specimen, respectively. CONCLUSION: Cytology using the Ayre spatula has low accuracy in detecting residual disease in the cervix post-LLETZ. PMID- 12078978 TI - A virus-like agent associated with neurofibromatosis in damselfish. AB - Damselfish neurofibromatosis (DNF) is a transmissible disease involving neurofibromas and chromatophoromas affecting bicolor damselfish Stegastes partitus on Florida reefs. Analysis of genomic DNA by Southern blotting techniques demonstrated the presence of a group of extrachromosomal DNAs in tumors from fish affected with DNF but not in healthy individuals. Cell lines obtained from tumors contained identical DNAs and were shown to be tumorigenic in vivo, while lines established from healthy fish did not contain such DNA and were not tumorigenic. These DNA patterns were also observed in experimentally induced tumors. A DNase resistant component of this DNA was isolated from both tumor cells and conditioned media of tumor cell lines suggesting that these sequences were encapsulated in viral particles. These data support the hypothesis that one or more of these extrachromosomal DNA forms is the genome of an unusual virus and that this virus is the etiologic agent of DNF. We have tentatively termed this agent the damselfish virus-like agent (DVLA). PMID- 12078977 TI - Posterior leukoencephalopathy following cisplatin, bleomycin and vinblastine therapy for germ cell tumor of the ovary. AB - A 31-year-old female developed multiple episodes of grand mal seizures after combination chemotherapy with cisplatin, vinblastine and bleomycin for germ cell ovarian cancer stage Ic. The clinicoradiologic features in this patient were consistent with posterior leukoencephalopathy, which is a rare complication of chemotherapy. Seizures were controlled by the anticonvulsive agent Dilantin (Pfizer, Khet Klongtoey, Bangkok) and she returned home without any permanent neurologic deficits. Follow-up magnetic resonance imaging 2 weeks later showed complete resolution of the abnormalities. This syndrome has been previously reported following cisplatin-based chemotherapy. Physicians should remain alert to the potential hazards of chemotherapy to the central nervous system. Risks and benefits should be seriously considered before starting treatment. PMID- 12078979 TI - Segment 8 encodes a structural protein of infectious salmon anaemia virus (ISAV); the co-linear transcript from Segment 7 probably encodes a non-structural or minor structural protein. AB - In this study we present the cloning, expression and partial identification of Genomic Segment 7 of infectious salmon anaemia virus (ISAV). The nucleotide sequence corresponding to Segment 7 was isolated from a bacteriophage lambda cDNA library and contained 2 overlapping open reading frames (ORFs) of 903 and 522 bases respectively. It also contained an ISAV-specific conserved nucleotide motif in the mRNA 5' region. The co-linear transcript representing the large ORF undergoes a splicing event that removes a 526 nucleotide intron to form a mRNA corresponding to the smaller reading frame. Thus, ISAV Genomic Segment 7 has a similar coding strategy as influenza A virus Segments 7 and 8. The largest ORF of Segment 7 and the first ORF of Segment 8 was expressed in E. coli as fusion proteins and rabbit antiserum was raised against the recombinant protein from Segment 8. Immunoblot studies using this antiserum and a serum against purified virus, show that Segment 8 encodes one of the major structural proteins of the virus whereas the co-linear ORF of Segment 7 probably encodes a non- or minor structural protein PMID- 12078980 TI - Detection of infectious salmon anemia virus in sea water by nested RT-PCR. AB - A method to detect low levels of infectious salmon anemia virus (ISAV) in environmental samples has been developed. The method is based on concentrating the viruses by tangential flow filtration and ultracentrifugation prior to amplification of the extracted viral RNA by nested reverse transcription polymerase chain reaction (RT-PCR). In the current investigation, seawater samples from salmon holdings were used for ISAV identification. ISAV was detected in seawater samples from a salmon holding site and from a vessel transporting salmon in 2 consecutive trials of the methodology. When known concentrations of ISAV were added to 21 of sea water, 5.5 viruses ml(-1) (corresponding to a tissue culture titer of TCID50 1.6 x 10(-2) ml(-1)) were detected using this method. PMID- 12078982 TI - PCR-based assays for the fish pathogen Aeromonas salmonicida. II. Further evaluation and validation of three PCR primer sets with infected fish. AB - Two Aeromonas salmonicida-specific polymerase chain reaction (PCR) tests and 1 A. salmonicida subsp. salmonicida-specific PCR test were used to screen salmonid populations that were either overtly or covertly infected with A. salmonicida subsp. salmonicida. It was demonstrated that these PCR assays could be used to replace the biochemical testing currently employed to confirm the identity of A. salmonicida isolates cultured from infected fish. The AP and PAAS PCR assays were also capable of direct detection of A. salmonicida in overtly infected fish, with mucus, gill and kidney samples most likely to yield a positive result. Culture was a more reliable method for the direct detection of A. salmonicida in covertly infected salmonids than was the direct PCR testing of tissue samples, with the AP and PAAS PCRs having a lower detection limit (LDL) of approximately 4 x 10(5) colony-forming units (CFU) g(-1) sample. PMID- 12078981 TI - PCR-based assays for the fish pathogen Aeromonas salmonicida. I. Evaluation of three PCR primer sets for detection and identification. AB - In an effort to develop a rapid diagnostic test for the fish pathogen Aeromonas salmonicida, the performance of 2 polymerase chain reaction (PCR) primer sets (AP and PAAS) targeting the fish pathogen A. salmonicida and 1 PCR primer set (MIY) targeting A. salmonicida subsp. salmonicida were evaluated. Initially, the PCR assays were used to screen purified DNA extracted from 308 A. salmonicida isolates. The AP and PAAS PCR tests were demonstrated to be 100% specific for the species A. salmonicida and did not cross-react with any of the non-target organisms (bacterial species other than A. salmonicida) used in this study. The combined sensitivity of the AP and PAAS tests was 99.4% and offered the best coverage in terms of identifying the target organism. The MIY PCR appeared to be 100% sensitive and specific for A. salmonicida subsp. salmonicida. Studies with tissues, spiked with known quantities of bacteria, were conducted to determine the lower detection limit of the PCR tests, and then the ability of these PCR tests to detect A. salmonicida in experimentally infected salmonids was assessed. PMID- 12078983 TI - Does size really matter? Effects of fish surface area on the settlement and initial survival of Lepeophtheirus salmonis, an ectoparasite of Atlantic salmon Salmo salar. AB - The effect of the size of the surface area of a fish host on settlement and initial survival of Lepeophtheirus salmonis (Kroyer, 1837) was determined. Atlantic salmon Salmo salar L. of various sizes-small (43 +/- 4 g), medium (173 +/- 32 g) and large (644 +/- 62 g)-were initially examined to ascertain their respective body surface area, excluding gill surface. The initial examination showed a size-dependent body to fin area ratio, with the fin area of small fish representing 34% of the total body area and the fin area of medium and large fish representing 26 and 23%, respectively. Regression analysis of the body weight and standard length against total body surface area gave a good correlation and high R2 values. Two simultaneous experimental infections with approximately 7,000 copepodids of L. salmonis were carried out on fish populations of mixed sizes consisting of 30 small fish, 10 medium-sized fish and 5 large fish, with an approximate total surface area per size group of 2700 cm2. Higher numbers of parasites were found on the small size group, which also had the highest parasite density, with 0.25 and 0.45 parasites cm(-2). Comparison of samples of 5 fish per experimental group revealed that the larger fish had the highest mean numbers of parasites, but the smaller fish still retained the highest density. There was a statistically significantly higher settlement on the fins than on the remainder of the body surface in all size groups. Highest numbers of parasites were particularly found on the dorsal and pectoral fins. Examination of the surface surface area revealed that the dorsal fin had the greater parasite density, with >2 parasites cm(-2) in all fish size groups. These data provide insight into the effects of the amount of host surface area available on parasite settlement and survival, and highlight the potentially increased susceptibility of farmed salmon smolts to infection of this ectoparasite. PMID- 12078984 TI - Occurrence of actinosporean stages of myxosporeans in an inflow brook of a salmon hatchery in the Mena River System, Hokkaido, Japan. AB - Actinospore infection of oligochaetes living in the mud and on the roots of vegetation in an inflow brook of a Hokkaido salmon hatchery was studied within the framework of a Japanese-Hungarian research program. Two triactinomyxon types, 1 echinactinomyxon, and 1 neoactinomyxum type were isolated from the oligochaete Rhyacodrilus komarovi Timm, 1990 collected during the survey. The aurantiactinomyxons were recorded over a period of 3 mo starting from the day after oligochaete collection. The oligochaetes released actinospores for several weeks from the first day of the study. Spore excretion of individual oligochaetes was not synchronous. Of the oligochaetes examined, 0.7, 7, 3 and 3%, were infected with the echinactinomyxon, neoactinomyxum and the 2 types of triactinomyxon spores, respectively. Actinospore infection was intense in the positive oligochaetes in all 4 types. Of the 4 actinospore types presented here, 3 are described for the first time. PMID- 12078985 TI - Dynamics of Kudoa camarguensis (myxosporean) infection in two gobiid species, Pomatoschistus microps and P. minutus (Teleostei: Pisces), in the Rhjne River delta, France. AB - The occurrence of the myxosporean parasite Kudoa camarguensis was surveyed monthly during 1997 in a brackish-water lagoon of the Rhjne River delta (France). K. camarguensis was found on its typical host, Pomatoschistus microps, and on an additional host, P. minutus. Prevalence and mean abundance were higher in the typical host than in the additional host due to differences in the temporal occupancy of the lagoon by the 2 species. The temporal occurrence of this myxosporean parasite is discussed in relation to the migratory habits of P. minutus and the sedentary habits of P. microps. PMID- 12078986 TI - Experimental challenge and clinical cases of Bohle iridovirus (BIV) in native Australian anurans. AB - Ranaviruses have been observed with increasing frequency amongst poikilothermic vertebrate hosts. The impact of ranaviruses upon amphibian populations has remained largely unknown. A gene probe for Bohle iridovirus (BIV) based upon primers designed to detect epizootic haematopoietic necrosis virus (EHNV) was constructed. A PCR and dot-blot system was used successfully in screening for the presence of BIV nucleic acid in digested formalin-fixed, paraffin-embedded amphibian tissues. Juvenile frogs were more susceptible to BIV than adults. In experimental challenges and epizootics in captive frogs, juvenile Litoria caerulea, L. alboguttata, Cyclorana brevipes and Pseudophryne coriacea were acutely susceptible. High mortality (at or near 100%) resulted, usually occurring within 5 to 25 d depending on dose and method of exposure. Histopathological changes included mainly hepatic, renal and splenic necroses. Significant haemosiderosis was encountered in more chronically infected frogs. BIV could be reisolated from juvenile L. caerulea >40 d after inoculation, and >200 d after the first mortalities occurred in an epizootic in L. alboguttata. Adult L. rubella, L. inermis, L. caerulea, Cophixalus ornatus and Taudactylus acutirostris were less susceptible in trials ranging from 30 to > 100 d. There was some evidence of chronic infection, and BIV could be detected by PCR. Wild moribund adult L. caerulea from Townsville and captive juvenile Pseudophryne corieacea from Sydney undergoing mortality tested positive with the BIV PCR. PCR and dot blot was more sensitive than viral isolation. PCR could detect BIV in amphibians long after BIV challenge, and in amphibians which appeared healthy. Ranaviruses could be having an impact on Australian herpetofauna. PMID- 12078987 TI - Low sequence variation among isolates of infectious hypodermal and hematopoietic necrosis virus (IHHNV) originating from Hawaii and the Americas. AB - A 2.9 kb fragment of the infectious hypodermal and hematopoietic necrosis virus (IHHNV) genome, which contains the coding sequence of putative non-structural and capsid proteins, was amplified and sequenced from each of 14 IHHNV isolates collected from cultured penaeid shrimp stocks in Hawaii and various sites in the Americas between 1982 and 1997. The sequence comparison indicates that the IHHNV genome is very stable, with 99.6 to 100% similarity among these 14 isolates. Only nucleotide substitutions were found. The percentage of substitution was higher in the putative capsid proteins region (1.3%) than in the putative non-structural proteins region (0.6%). Out of 25 substitutions found, 14 resulted in amino acid changes. There is no apparent association between clinical outcomes and particular amino acid substitutions. Based on genetic distances, the isolates were clustered into 3 groups that generally correspond with their geographic origins. PMID- 12078988 TI - First report of piscine nodavirus infecting wild winter flounder Pleuronectes americanus in Passamaquoddy Bay, New Brunswick, Canada. AB - Piscine nodaviruses (Betanodaviridae) are frequently reported from a variety of cultured and wild finfishes. These non-enveloped, single-stranded RNA virions cause viral encephalopathy and retinopathy (VER), also known as viral nervous necrosis (VNN) or fish encephalitis. Recently, nodavirus infections have posed serious problems for larval and juvenile cultured halibut Hippoglossus hippoglossus in Norway and Scotland. To date, no such viruses have been described from any cultured or wild pleuronectid in Atlantic Canada. Obviously, there exists a need to survey wild populations of pleuronectids to assess the risk of potential transfer of nodavirus from wild to caged fishes. This paper presents the results of monthly surveys (April 2000 to March 2001) of viruses from wild winter flounder Pleuronectes americanus collected from Passamaquoddy Bay, New Brunswick, Canada. Tissue samples from wild flounder were screened initially on commercial cell lines (EPC, SSN-1, SHK and CHSE-214) for any evidence of cytopathic effect (CPE). After confirmation of CPE, nodavirus identification was achieved using reverse transcription polymerase chain reaction (RT-PCR) analysis. We detected nodavirus from only 1 out of 440 flounder (0.23%) examined. This is the first report of piscine nodavirus isolated from wild winter flounder in Atlantic Canada, and although this prevalence may seem low, we discuss the implications of this finding for Canada's emerging halibut aquaculture industry. PMID- 12078989 TI - Disordered drug delivery: destiny, dynamics and the Deborah number. AB - "Disordered drug delivery" is an innovative approach to improving the performance of new chemical entities delivered to the human body. In this technique, the molecules of the drug and/or its delivery system are kinetically trapped in a high energy non-crystalline state. The resulting disordered or "amorphous" material offers potential increases in solubility and biological activity of many thousand fold compared with more conventional crystalline forms of the drug. Despite having a molecular level structure akin to that of liquids, amorphous materials have macroscopic properties that are typical of solids and thus they may be presented to the patient in the form of a convenient solid dosage form. Significant advances in the fundamental understanding of amorphous pharmaceutical materials in the past ten years have permitted major steps forward in the rational design of disordered drug delivery systems. Recognition of significant levels of molecular mobility in the glassy regime and an in-depth appreciation of molecular relaxation times and their distributions have enabled rapid progress to be made in this field. Needs for the future include analytical techniques that can elucidate the complex, dynamic and heterogeneous structure of amorphous materials and reliable models to predict the physical stability and in-vivo performance of disordered drug delivery systems. There are also significant opportunities for the production of disordered drug delivery systems with tailor made properties through the careful engineering of custom amorphous alloys. PMID- 12078990 TI - In-vitro release of bupivacaine from injectable lipid formulations investigated by a single drop technique--relation to duration of action in-vivo. AB - The aim of this study was to develop an in-vitro release method suitable for injectable slow-release lipid formulations of local anaesthetics (or other drugs). We also aimed that the results of the in-vitro measurements should have a clear relationship to duration of action in-vivo. Six formulations of bupivacaine base in medium-chain triglyceride-glyceryl dilaurate mixtures were developed. A new apparatus was constructed for determination of their in-vitro release profiles. A bulbous glass tube was fixed inside a standard glass bottle, which was then filled with release medium. A stirring magnet was enclosed in the perforated polypropylene cylinder holding the glass tube. The stirring created a continuous, rotating downward flow of medium inside the tube, which kept the lipid phase, introduced by means of a syringe, suspended as a single, free drop. Release profiles were obtained by sampling of the release medium for up to 72 h and analysis by gas-liquid chromatography. The duration of action in-vivo of the respective formulations was tested by the hot-plate method in rats. The release profiles of bupivacaine in-vitro were mono-exponential for four formulations and bi-exponential for the other two. There was a positive correlation between the proportion of glyceryl dilaurate in the formulation and the slow half-life of release of bupivacaine. All formulations showed prolonged duration of action in vivo, median values within the range 4.5-12 h, as compared with a 2-h effect of bupivacaine hydrochloride solution. A comparison of in-vitro release curves and durations of action in-vivo suggested that to maintain nerve blockade in-vivo the formulations must release bupivacaine at a rate of approximately 350 microg h(-1) under the in-vitro conditions. To conclude, we designed and tested a novel apparatus for measuring release of a local anaesthetic (or other drug) from a fluid or semi-solid formulation in-vitro. Release rates obtained in-vitro by means of this technique may be used to guide the development of formulations with suitable durations of action in-vivo. The apparatus is, however, as yet a prototype. Rigorous evaluation of performance should be carried out on devices built to specific standards according to their intended application. PMID- 12078991 TI - In-vitro release of fluoropyrimidines from PLGA film implants. AB - The release of two low-molecular weight water-soluble fluoropyrimidines, 5 fluorouracil and 5-fluorouridine, from implants of PLGA films was modulated by varying the area (diameter) and number of layers of film per implant. The aim was to achieve continuous release without burst effect for at least a month. The film implants were prepared by the solvent evaporation technique. Except with 5 fluorouracil films, the in-vitro release profiles were in all cases triphasic, indicating that release proceeds by a combination of diffusion and polymer erosion. The experimental data fit the equation resulting from the sum of two exponentials, one direct and the other inverse. 5-fluorouridine release from simple films presented a relatively minor burst effect (24-28%). In contrast, the delivery of both compounds from sandwich-type implants occurred continuously without a burst effect, and lasted for 17-20 days. During the first phase, both 3 and 5-mm sandwiches released 55% of the dose of 5-fluorouridine, at rate constants of 0.037+/-0.021 h(-1) (n = 3) and 0.009+/-0.003 h(-1) (n=3), respectively. In the second phase, release was gradual from both simple films (k2 = 0.011-0.015 h(-1)) and sandwiches (k2 = 0.018-0.058 h(-1)). According to the analysis-of-variance results, neither the area nor type of implant influenced the rate constants significantly. The release profiles of 5-fluorouracil from simple films showed a severe burst effect (64-71%). Release of 5-fluorouracil was gradual only from sandwiches, 5 mm in diameter, showing a lag time unobserved in the 3-mm sandwiches. In the second phase, release was gradual (k2 = 0.014+/-0.003 h(-1)) from 3-mm implants. However, the high variability in results for 5-mm implants prevents conclusions being drawn about the model parameters. Therefore, the sandwich-type film implants showed their utility for releasing water-soluble drugs for a prolonged time, without burst effect. PMID- 12078992 TI - Use of a cholesterol-rich emulsion that binds to low-density lipoprotein receptors as a vehicle for paclitaxel. AB - A cholesterol-rich emulsion (LDE) is taken up by malignant cells which over express low-density lipoprotein (LDL) receptors and thus may be used as a carrier for drugs directed against neoplastic cells. In this study, we associated the antineoplastic agent paclitaxel to LDE and analysed the new formulation's incorporation efficiency, chemical and physical stability, cellular uptake and cytostatic activity against a neoplastic cell line and the acute toxicity to rats. A paclitaxel incorporation efficiency of approximately 75% was achieved when paclitaxel was mixed with LDE at a 6:1 lipid-to-drug molar ratio. The association of paclitaxel with LDE increased by 54% the mean diameter of the emulsion particles but did not damage the paclitaxel chemical structure as analysed by HPLC. Results from gradient ultracentrifugation and Sephadex G25 gel filtration indicated that the binding of the drug to the emulsion was stable. It was shown that the cellular uptake and the cytotoxic activity of LDE-paclitaxel by a neoplastic cell line (NCI-H292 cells) was indeed mediated by the LDL receptors. The antiproliferative activity of LDE-paclitaxel against NCI-H292 cells was less than that of a commercial paclitaxel preparation (50% inhibitory concentration, IC50 = 2.60 and 0.45 microM, respectively). This difference, however, can be ascribed to the in-vitro anti-proliferative activity of the commercial paclitaxel vehicle Cremophor EL; when Cremophor EL was added to the cultures with LDE-paclitaxel, the IC50 value was reduced to 0.45 microM, attaining that of the commercial paclitaxel preparation. The tolerability of LDE paclitaxel in rats was remarkable, such that its lethal dose (LD50) was ten-fold greater than that of the commercial formulation (LD50 = 324 and 31.8 mg kg(-1), respectively). Therefore, LDE-paclitaxel association is stable and the cytostatic activity of the drug is preserved while its toxicity to rats is small. By diminishing the side effects and directing paclitaxel to neoplastic tissues, LDE may be useful as adjuvant in chemotherapy with this drug. PMID- 12078993 TI - Interaction of double-chained cationic surfactants, dimethyldialkylammoniums, with erythrocyte membranes: stabilization of the cationic vesicles by phosphatidylcholines with unsaturated fatty acyl chains. AB - We studied the interaction of double-chained cationic surfactants, dimethyldialkylammoniums, (CH3)2N+(CnH2n+1)2, with the lipid bilayer of guinea pig erythrocytes by observing the haemolysis, aggregation and shape change in the erythrocytes. In the presence of sonicated dispersions of the five dimethyldialkylammoniums tested (n = 10, 12, 14, 16 and 18), haemolysis was induced dose dependently, and at 0.1 mM or higher concentrations, haemolysis was induced more rapidly by dimethyldialkylammoniums with shorter alkyl chains. The cationic surfactants with longer alkyl chains, such as dimethyldipalmitylammonium, induced aggregation of the erythrocytes before haemolysis fully progressed. The vesicles of these long-chain dimethyldialkylammoniums in the presence of phosphatidylcholines with unsaturated fatty acyl chains markedly reduced the haemolysis rates. Furthermore, in the presence of phosphatidylcholines with unsaturated acyl chains the formation of tightly aggregated structures of several erythrocytes was observed. These findings, and analysis by spin label 5-doxylstearic acid, indicate that phosphatidylcholines enriched with unsaturated acyl chains stabilize the cationic vesicles of long-chain dimethyldialkylammoniums and the interaction with the lipid bilayer of erythrocyte membranes as cationic vesicles became prominent. PMID- 12078994 TI - Introduction of poly-L-lactic acid microspheres into the skin using supersonic flow: effects of helium gas pressure, particle size and microparticle dose on the amount introduced into hairless rat skin. AB - A microparticulate bombardment system loaded with DNA- and RNA-coated gold and tungsten microparticles (diameter 1-3 microm; density about 19 g cm(-3)), the Helios gene gun system (Helios gun system), has been used to deliver a gene into cells by accelerating the microparticles to high velocity using a supersonic flow of helium gas. To investigate whether drug-loaded microspheres, > 20 microm in diameter and about 1.0 g cm(-3) in density, could be delivered in powder form quantitatively into the skin using the Helios gun system equipped with a cartridge container fitted with a rupture membrane, we investigated the effect of the helium gas pressure in accelerating indometacin-loaded poly-L-lactic acid (PLA) microspheres, as well as the particle size and the bombardment dose on delivery into the skin. Introduction of indometacin (i.e. indometacin-loaded PLA microspheres) after bombardment, with 3.0 mg indometacin-loaded PLA microspheres of a particle size of 20-38, 44-53 and 75-100 microm at a helium pressure of 100, 200 and 300 psi, of the abdomen of hairless rats increased in parallel with the helium pressure and it was also affected by the particle size, being highest at a diameter of 75-100 microm. However, introduction of higher amounts of PLA microspheres resulted in more severe skin erythema (skin damage) as monitored by the Draize score. Using lower bombardment doses (0.5 and 1.0 mg), the efficiency of introduction was improved and the skin damage markedly reduced. Moreover, discrete bombardment with a low dose provided a more efficient introduction of indometacin and less skin damage. These results suggest that bombardment injection of drug-loaded microspheres in a powdered form by the Helios gun system appears to be a very useful tool for the quantitative delivery of a variety of drugs and an alternative to parenteral injection by needle, especially for delivering water-soluble macromolecules. PMID- 12078995 TI - Pharmacokinetic-pharmacodynamic modelling of insulin: comparison of indirect pharmacodynamic response with effect-compartment link models. AB - The pharmacokinetic and pharmacodynamic modelling of insulin has been reported using a combined pharmacokinetic/pharmacodynamic (PK/PD) model, in which a hypothetical effect compartment is linked to a pharmacokinetic compartment. Review of the literature, however, indicated that the recently developed PK/PD models have consisted of an indirect pharmacodynamic response component, but none of them has been applied to the modelling of insulin. To study the relative relevance of the indirect pharmacodynamic response model and the effect compartment link model in modelling the pharmacokinetics and pharmacodynamics of insulin, regular human insulin was administered intravenously at a dose of 0.1 IU kg(-1) to healthy Yucatan minipigs (after an overnight fasting). The plasma concentrations of insulin were measured by radioimmunoassay at predetermined time intervals, while blood glucose levels were monitored continuously using a glucose monitor. Analysis of the plasma insulin and the blood glucose profiles was performed by fitting with various PK/PD models and the results indicated that all of the 12 sets of plasma insulin data (after normalizing by the basal levels) have been adequately fitted to the two-compartment open pharmacokinetic model (a mean+/-s.e. correlation coefficient of 0.996+/-0.001 was obtained). The mean+/ s.e. correlation coefficient, the weighted residuals sum of squares (WRSS), and the Akaike's information criterion (AIC) were found, respectively, to be 0.935+/ 0.008, 624+/-67, and 522+/-9 for the inhibitory indirect pharmacodynamic response model and 0.941+/-0.010, 547+/-63 and 513+/-9 for the stimulatory indirect pharmacodynamic response model, as compared with 0.725+/-0.041, 2309+/-276 and 628+/-10 for the effect-compartment link model. Based on these results, one may conclude that the indirect pharmacodynamic response model is a more appropriate approach for modelling the PK/PD of insulin than the effect-compartment link model. PMID- 12078996 TI - Clearance and disposition of indometacin in chronically instrumented fetal lambs following a 3-day continuous intravenous infusion. AB - Indometacin is used in pregnancy for the treatment of premature labour, but there are limited data on the disposition of the drug in the fetus. In order to elucidate fetal indometacin pharmacokinetics at plasma levels and duration comparable with those occurring with use of the drug for tocolysis in humans, indometacin was administered at doses of 1.9 (low dose, LD; n = 5) or 7.5 (high dose, HD; n = 9) microg min(-1) to steady state over a 3-day period in chronically instrumented fetal lambs. Indometacin concentrations in biological fluid samples were analysed by a sensitive capillary gas chromatography-electron capture detection method. The mean steady-state fetal arterial plasma indometacin concentrations were 68.6+/-16.5 ng mL(-1) in the LD infusion and 230.3+/-28.8 ng mL(-1) in the HD infusion. Indometacin concentrations in amniotic fluid were approximately 10% of those in fetal plasma, and below assay detection limits in tracheal fluid. Total body clearance (TBC) in the LD and HD infusions were not different and the overall mean was 11.3+/-1.2 mL min(-1) kg(-1). In the 11 experiments where paired fetal arterial and umbilical venous samples were collected, the extraction of indometacin across the placenta averaged only 5.2+/ 1.1%, indicating low placental permeability to the drug in sheep. However, fetal placental clearance (CLpl) of indometacin (10.0+/-2.5 mL min(-1) kg(-1), n = 10) averaged 115.1+/-41.2% of TBC in these animals and the calculated value for fetal non-placental clearance (0.6+/-2.8 mL min(-1) kg(-1)) was not significantly different from zero. Fetal renal clearance of intact indometacin (3.8+/-1.1 microL min(-1) kg(-1); n = 12) was also very low. However, treatment of fetal urine with glucuronidase indicated the presence of glucuronide conjugates and these comprised 69.9+/-8.2% of the total drug concentration (i.e. intact+conjugated) in urine. Thus, the fetal lamb appears to be able to glucuronidate indometacin, but the contribution of this and other non-placental routes to overall fetal elimination of the drug appear minimal. CLpl of the drug is also low owing to the physicochemical properties of indometacin (high polarity) and the permeability characteristics of the sheep placenta. PMID- 12078997 TI - Effect of dietary fatty acids on the intestinal permeability of marker drug compounds in excised rat jejunum. AB - The aim of this study was to explore the effects of diets containing saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and omega-3 and omega-6 polyunsaturated fatty acids (omega-3 and omega-6 PUFA, respectively) on the passive and active transport properties of rat jejunum using marker compounds. Rats were fed diets supplemented with 18.4% (w/w) lipid (4 groups) or standard rat chow (1 group) for a period of 30 days. At the end of the dietary period, mucosal scrapings were taken for the determination of membrane phospholipids, and the apparent jejunal permeability of radiolabelled marker compounds was determined using modified Ussing chambers. Changes in the phospholipid content of the brush border membrane reflected the different lipid content of the diets. The passive paracellular permeability of mannitol was not significantly affected by the fatty acid composition of the diet, although there was a trend toward decreased mannitol permeability in the rats fed both the omega-3 and omega-6 PUFA diets. In comparison, the transcellular diffusion of diazepam was reduced by 20% (P < 0.05) in rats fed diets supplemented with omega-3 and omega-6 PUFA. In the lipid-fed rats, the serosal to mucosal flux of digoxin, an intestinal P glycoprotein substrate, was reduced by 20% (P < 0.05) relative to the chow-fed group, however there were no significant differences between the different lipid groups. The active absorption of D-glucose via the Na+-dependent transport pathway was highest in the SFA, MUFA and PUFA omega-3 dietary groups, intermediate in the low-fat chow group and lowest in the PUFA omega-6 group, and was positively correlated with short-circuit current. These studies indicate that dietary fatty acid changes can result in moderate changes to the active and passive transport properties of excised rat jejunum. PMID- 12078998 TI - Influence of nicardipine and nifedipine on plasma carvedilol disposition after oral administration in rats. AB - The effect of two kinds of 1,4-dihydropyridine calcium-channel blockers, nicardipine hydrochloride and nifedipine, on the disposition of carvedilol, was studied in rats. Blood samples were assayed for carvedilol levels using solid phase extraction and high-performance liquid chromatography. The plasma carvedilol concentration was found to be significantly higher, and the area under the concentration-time curve up to 24 h (AUC0-->24) was 6.7 and 3.0 times higher after simultaneous oral administration of 20 mg kg(-1) carvedilol with 40 mg kg( 1) nicardipine hydrochloride, or with 40 mg kg(-1) nifedipine, respectively, than after administration of carvedilol alone. The pharmacokinetic interaction between carvedilol and dihydropyridine calcium-channel blockers is thought to be attributable to vasodilator-induced changes in hepatic first-pass metabolism, inhibition in the absorption barrier by P-glycoprotein and in the metabolism of carvedilol. PMID- 12078999 TI - Synthesis and evaluation of N1-substituted-3-propyl-1,4-benzodiazepine-2-ones as cholecystokinin (CCK2) receptor ligands. AB - A novel synthetic approach towards N1-alkylated 3-propyl-1,4-benzodiazepines was developed in five synthetic steps from 2-amino-4-chlorobenzophenone, in which the N-oxide 4 served as a key intermediate. The structure-activity relationship optimization of this 3-propyl-1,4-benzodiazepine template was carried out on the N1-position by selective alkylation reactions and resulted in a ligand with an improved affinity on the cholecystokinin (CCK2) receptor. The N-allyl-3-propyl benzodiazepine 6d displayed an affinity towards the CCK2 (CCK-B) receptor of 170 nM in a radiolabelled receptor-binding assay. The anxiolytic activity of this allyl-3-propyl-1,4-benzodiazepine 6d was subsequently determined in in-vivo psychotropic assays. This novel ligand had ED50 values of 4.7 and 5.2 mg kg(-1) in the black and white box test and the x-maze, respectively, and no significant sedation/muscle relaxation was observed. PMID- 12079000 TI - Free radical scavengers, anti-inflammatory and analgesic activity of Acaena magellanica. AB - Extracts of the whole plant Acaena magellanica (Rosaceae) were assessed for anti inflammatory, antipyretic and analgesic activity in animal models. At 600 mg kg( 1), the global ethanolic extract (GEE), dichloromethane (DCM) and defatted methanol (MeOH) fractions showed a mild anti-inflammatory effect in the carrageenan-induced guinea-pig paw oedema. The GEE, DCM and defatted MeOH fractions significantly reduced inflammation by 43.2, 40.5 and 42.1%, respectively. The GEE did not showed any significant antipyretic activity in doses up to 600 mg kg(-1). A 20% w/v infusion administered orally at 16 mL kg(-1) presented analgesic effect in the acetic acid-induced abdominal constriction test in mice. The GEE and MeOH extract of A. magellanica showed free radical scavenging activity in the diphenylpicrylhydrazyl decolouration assay. Assay guided isolation led to quercetin, Q-3-O-beta-glucoside, Q-3-O-beta-D galactoside, ellagic acid and catechin as the free radical scavengers. The saponins tormentic acid 28-O-beta-D-galactopyranoside and 28-O-beta-D glucopyranoside were isolated from the polar extract. The structures were determined by spectroscopic methods. PMID- 12079001 TI - Mechanisms involved in the relaxant action of the ethanolic extract of propolis in the guinea-pig trachea in-vitro. AB - This study examines the mechanisms by which the standardised ethanolic extract of propolis induces relaxation of the guinea-pig trachea in-vitro. In guinea-pig trachea with or without epithelium and contracted by histamine, the propolis extract caused reproducible and graded relaxation, with a mean EC50 value of 3.8 or 10.5 microg mL(-1) and Emax of 100%, respectively. The propolis extract induced relaxation was markedly reduced (26+/-9 and 96+/-3%) when guinea-pig tracheas were exposed to Krebs solution containing elevated K+ in the medium (40 or 80 mM). Pre-incubation of guinea-pig tracheas with tetraethylamonium (100 mM) or with 4-aminopyridine (10mM) reduced the propolis extract-induced relaxation by 31+/-10% and 28+/-2%. Likewise, apamin (0.1 microM), charybdotoxin (0.1 microM) or iberiotoxin (0.1 microM) caused marked inhibition of propolis extract-mediated relaxation in guinea-pig trachea (percentage of inhibition: 65+/-3%, 60+/-5% and 65+/-9%, respectively). Also, glibenclamide (1 microM) inhibited the relaxant response caused by the propolis extract by 57+/-4%. Omega-conotoxin GIVA (0.1 microM) or capsaicin (1 microM) produced small but significant inhibition (30+/ 5% or 47+/-7%, respectively) of the propolis extract-induced relaxation. The vasoactive intestinal peptide (VIP) antagonist D-p-Cl-Phe6,Leu17[VIP] porcine (0.1 microM) inhibited relaxation by 55+/-5%, while propranolol (1 microM) induced a parallel rightward displacement (about 20 fold) of the propolis extract concentration-response curve. Finally, the propolis extract-induced relaxation was inhibited by the nitric oxide synthase inhibitor L-N(G)-nitroarginine (L NOArg, 100 microM) (48+/-6%), and by the soluble guanylatecyclase inhibitormethylene blue (10 microM) (37+/-6%), whilethe moreselectivesoluble guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolol[4,3-alquinoxalin-1-one (ODQ, 1 microM) produced only a parallel (about 3 fold) rightward displacement of the propolis extract concentration-response curve. Collectively, these results support the notion that the propolis extract-mediated relaxation in the guinea pig trachea involves the release of nitric oxide, probably from sensory neurons, besides the activation of soluble guanylate cyclase and activation of Ca2+- and ATP-sensitive K+ channels. Furthermore, the stimulation of beta2-adrenergic and VIP receptors also seems to account for its relaxant action. PMID- 12079002 TI - Mechanism of the vasodilator effect of 12-O-methylcurine in rat aortic rings. AB - The vasodilator effects of 12-O-methylcurine (OMC), a bisbenzylisoquinoline alkaloid isolated from Chondrodendron platyphyllum (Menispermaceae), and its respective mechanism of action were investigated in rat aorta. In either endothelium-intact or endothelium-denuded aortic rings, OMC induced concentration dependent relaxation in vessels pre-contracted with 0.1 microM phenylephrine (IC50 = 63.2+/-8.8 microM and 73.9+/-5.3 microM, respectively), 100 microM 5 hydroxytryptamine (IC50=49.6+/-13 microM and 49.9+/-10 microM, respectively) and 50 mM KCl (IC50= 19.9+/-6.8 microM and 21.1+/-4.5 microM, respectively). OMC also inhibited in a concentration-dependent and non-competitive manner the concentration-response curves induced by CaCl2 in high K+ (IC50 = 16.7+/-1.6 microM). In addition, OMC (100 microM) strongly inhibited phenylephrine-induced contractions dependent on calcium influx in the absence and presence of nifedipine (10 microM). In Ca2+-free medium, the transient contractions induced by phenylephrine (0.1 microM) were strongly inhibited by OMC (100 microM), whereas those induced by caffeine (20 mM) were not altered. H-89 (1 microM) and Rp-8-pCPT-cGMPs (3 microM), selective inhibitors of protein kinase A and G, respectively, did not change the relaxant effect of OMC in aortic rings pre contracted with phenylephrine. Finally, OMC induced a concentration-dependent relaxation (IC50 = 62.8+/-12.5 microM) of the sustained contractions induced by the phorbol ester 12-O-tetradecanoyl phorbol-13-acetate in normal, but not in Ca2+-free, solution. The above results suggest that OMC induces a vasodilator effect in rataortic rings by a mechanism independent of the presence of functional endothelium and dependent on the influx of calcium ions through voltage- and receptor-operated calcium channels. Furthermore, it can also be suggested that the inhibition of calcium influx activated by protein kinase C is involved in the vasodilator effect of OMC. PMID- 12079004 TI - Effect of nizatidine on paracetamol and its metabolites in human plasma. AB - The effect of the histamine H2-receptor antagonist, nizatidine, on plasma concentrations of paracetamol has been investigated with respectto hepatic metabolism. Paracetamol (1000 mg) together with 300 or 150 mg nizatidine or placebo was orally administered to five healthy male volunteers. Venous blood samples were taken before and after administration. Plasma paracetamol and paracetamol conjugates (glucuronide and sulfate) were measured by high performance liquid chromatography. The pharmacokinetic parameters were calculated from the plasma paracetamol concentration-time curves of each volunteer. The plasma nizatidine concentration was highest (2420.0+/-192.4 and 996.0+/-54.6 ng mL(-1)) in the sample taken 1 h after administration of 300 mg nizatidine (high dose) and 150mg nizatidine (low dose), respectively. Plasma paracetamol concentrations with nizatidine (high and low doses) were increased significantly at 45-120 min and 45-60 min, respectively, compared with placebo. The total area under the plasma paracetamol concentration-time curve from 0 to 180 min (2361.5+/ 146.4 and 2085.75+/-73.5 microg min mL(-1)) significantly increased after coadministration of nizatidine (high and low doses), respectively (P < 0.01 vs placebo). Paracetamol glucuronide concentrations with nizatidine (high and low doses) were decreased significantly at 30-45 min and 30 min, respectively, compared with placebo. However, plasma paracetamol sulfate concentrations with nizatidine (high and low doses) were not significantly altered. The coadministration of nizatidine (150 and 300 mg) dose-dependently reduces plasma paracetamol glucuronide concentrations and increases plasma paracetamol concentrations. The effects of nizatidine could result from the inhibition of glucuronyltransferase. Thus, care is necessary when paracetamol and nizatidine are coadministered. PMID- 12079003 TI - Effects of adenosine receptor agonists on guinea-pig isolated working hearts and the role of endothelium and NO. AB - The hypothesis that the coronary vasodilator effects of adenosine receptor agonists are independent of the vascular endothelium or mediators derived therefrom was examined in guinea-pig isolated working hearts. Adenosine receptor agonists, 5'-(N-ethylcarboxamido)-adenosine (NECA; two-fold selective for A2 over A1 receptors), 2-[p-(2-carboxyethyl)phenylethylamino]-5'-N ethylcarboxamidoadenosine (CGS21680; A2A selective), N6-cyclopentyl-adenosine (CPA; A1 selective) and N6-(3-iodobenzyl)adenosine-5'-N-methyluronamide (IB-MECA; A3 selective), were infused (3 x 10(-7) M) after endothelium removal by passing oxygen through the coronary circulation. In spontaneously beating hearts, CGS21680 and NECA increased, while CPA decreased, coronary flow. NECA and CPA reduced heart rate, left ventricular pressure and aortic output. The nitric oxide synthase (NOS) inhibitor, N(G)-nitro-L-arginine (L-NOARG; 3 x 10(-5) M) abolished the vasodilatation by NECA but not CGS21680, indicating that nitric oxide (NO) of a non-endothelial source mediated the NECA response. Coronary vasodilatation by CGS21680 was inhibited bythe A2A receptor antagonist, 4-(2-[7-amino-2-(2 furyl)[1,2,4]triazolo [2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol (ZM241385). Indometacin (10(-6) M) attenuated the coronary vasodilatation to CGS21680, suggesting a partial role for cyclooxygenase products. IB-MECA had no effect, indicating no A3 receptor involvement. In paced working hearts, the responses were similar except CPA had no effect on coronary flow or aortic output and CGS21680 increased left ventricular pressure and the maximum rate of ventricular pressure rise. This study has demonstrated functionally effective removal of the endothelium by a novel method of passing oxygen through the coronary vasculature. A coronary vasodilator action of adenosine receptor agonists mediated via A2A receptors is endothelium- and NO-independent, but partially involves cyclooxygenase products. PMID- 12079005 TI - Reversal of cannabinoids (delta9-THC) by the benzoflavone moiety from methanol extract of Passiflora incarnata Linneaus in mice: a possible therapy for cannabinoid addiction. AB - The newly reported benzoflavone moiety from the plant Passiflora incarnata Linneaus has been evaluated in light of traditional reports on the use of P. incarnata in breaking down cannabis addiction. In the modern or allopathic system of therapeutics, there has been no suitable remedy to combat the severe withdrawal effects of various cannabis products, including marihuana, marijuana, bhang, hashish, ganja, etc., the world-wide consumption of which has attained alarming proportions especially among the younger generation. Mice were given a 10-mg-kg(-1) twice-daily dose of delta9-tetrahydrocannabinol (delta9-THC) by mouth for six days to make them dependent upon cannabinoids. Concurrently, other groups of mice were administered delta9-THC along with a 10- or 20-mg-kg(-1) twice-daily dose of the benzoflavone moiety from P. incarnata orally for 6 days. Upon measuring locomotor activity during the treatment regimen, it was noticed that the mice receiving the P. incarnata extract and delta9-THC together developed significantly less tolerance and dependence, relative to the mice receiving delta9-THC alone. Upon administration of SR-141716A, a selective cannabinoid-receptor antagonist (10 mg kg(-1), p.o.) to all the groups of mice on the 7th day, an artificial withdrawal was produced due to an abrupt decline of delta9-THC levels in mouse brain. However, the typical withdrawal effects like paw tremors and head shakes were significantly less in the mice given delta9 THC+P. incarnata benzoflavone moiety for 6 days. Upon administration of 20 mg kg( 1) of the P. incarnata benzoflavone moiety to mice showing severe symptoms of withdrawal due to administration of SR-141716A, there was a marked attenuation of withdrawal effects, thereby suggesting the usefulness of the benzoflavone moiety in delta9-THC withdrawal. Thus, the benzoflavone moiety of P. incarnata, when administered concurrently with delta9-THC, prevented the development of tolerance and dependence of cannabinoids in mice. Even an acute administration of the benzoflavone moiety (20 mg kg(-1), p.o.) significantly blocked the expression of withdrawal effects in delta9-THC-dependent mice. PMID- 12079006 TI - Early failures among 7,174 primary total knee replacements: a follow-up study from the Norwegian Arthroplasty Register 1994-2000. AB - We studied primary total knee replacements (TKRs), reported to the Norwegian Arthroplasty Register, operated on between 1994 and 2000. A Cox multiple regression model was used to evaluate differences in survival among the prosthesis brands, their types of fixation, and whether or not the patella was resurfaced. In Norway in 1999, the incidence of knee prosthesis operations was 35 per 100,000 inhabitants. Cement was used as fixation in 87% of the knees, 10% were hybrid and 2% uncemented implants. Bicompartmental (not resurfaced patella) prostheses were used in 65% of the knees. With all revisions as endpoint, no statistically significant differences in the 5-year survival were found among the cemented tricompartmental prostheses brands: AGC 97% (n 279), Duracon 99% (n 101), Genesis I 95% (n 654), Kinemax 98% (n 213) and Tricon 96% (n 454). The bicompartmental LCS prostheses had a 5-year survival of 97% (n 476). The type of meniscal bearing in LCS knees had no effect on survival. Survival with revision for all causes as endpoint showed no differences among types of fixation, or bi- or tricompartmental prostheses. Pain alone was the commonest reason for revision of cemented bicompartmental prostheses. The risk of revision because of pain was 5.7 times higher (p < 0.001) in cemented bicompartmental prostheses than cemented tricompartmental ones, but the revisions mainly involved insertion of a patellar component. In tricompartmental prostheses the risk of revision because of infection was 2.5 times higher than in bicompartmental ones (p = 0.03). Young age (< 60) and the sequelae after a fracture increased the risk of revision. The 5 year survival of the 6 most used cemented tricompartmental knee prostheses brands varied between 95% and 99%, but the differences were not statistically significant. There were more revisions because of pain in bicompartmental than in tricompartmental knees. In tricompartmental knees, however, there were more revisions because of an infection. The relatively few patients with uncemented and hybrid implants showed no improvements in results compared to cemented knee prostheses. PMID- 12079007 TI - Degeneration of the atlanto-axial joints: a histological study of 9 cases. AB - Degeneration of the lateral atlanto-axial joints (AAJ) has been described as a potential cause of severe neck pain. However, hardly any data are available on its incidence, especially in comparison to the lower cervical spine. In this histological study, we examined the AAJs in 9 specimens from elderly patients, graded the findings and compared them to those in the facet joints of the lower cervical spine. Most histological changes in the AAJs were mild, but the changes in the lower cervical spine were severer. Previous mechanical studies have described the AAJ as a very mobile joint with large neutral zones, which may explain the mild degree of osteoarthrosis found in these specimens. PMID- 12079008 TI - Fixation of mid-third clavicular fractures with knowles pins: 78 patients followed for 2-7 years. AB - From January 1993 to December 1997, we treated 78 (75 fresh, 1 delayed union and 2 nonunion) fractures of the mid-third clavicle with open reduction and internal fixation using 3.8 mm Knowles pins. All patients used an arm sling for 2-6 weeks after surgery, and were told to avoid vigorous exercise or work with heavy loads for the next 6 weeks. 73 fractures healed within 6 months. 3 complications occurred relating to the surgical procedures. The final rate of bone union was 77/78, although 3 patients showed a delay in bone union and 1 fracture was revised because of a loosened Knowles pin. After a mean follow-up of 49 (24-86) months, the average shoulder function, as evaluated by a modified Constant-Murley score, was 97% versus the contralateral shoulder. We suggest that if surgery is indicated, open reduction and internal fixation using a Knowles pin is an effective method for managing mid-third clavicular fractures. PMID- 12079009 TI - Unrecognized shoulder joint penetration during fixation of proximal fractures of the humerus. AB - In osteosynthesis of proximal humerus fractures, the screws frequently penetrate the joint. Using orthogonal frontal and lateral radiographs, we found unrecognized pin penetration in 8 of 30 patients during fixation of unstable neck fractures of the humerus. PMID- 12079010 TI - Progressive surgical release of a posttraumatic stiff elbow. Technique and outcome after 2-18 years in 46 patients. AB - We treated 46 consecutive patients (47 elbows) suffering from posttraumatic contracture of the elbow joint with operative release. A lateral approach was used to perform a capsulectomy after release of the extensor muscles in 23 elbows. An additional medial approach was used to excise ulnar adhesions and perform a more extensive capsulectomy and an ulnar nerve neurolysis in 24 elbows. Postoperative rehabilitation consisted of immediate passive range-of-motion exercises. The results were assessed after average 10 (2-18) years. Before surgery, the mean active arc of motion was 45 (SD 36) degrees, which improved to 99 (SD 34) degrees after release. 7 patients had transient ulnar paresthesia, 4 recurrent stiffness, which did not become worse after surgery, 1 was reoperated on and 2 developed a postoperative infection, which responded to antibiotic treatment after a few months. No patient suffered from joint instability, or an increase in pain. 44 patients were satisfied with the result at the latest follow up. We found similar improvement in both treatment groups. PMID- 12079011 TI - Patients with laxity of the distal radioulnar joint after distal radial fractures have impaired function, but no loss of strength. AB - Laxity of the distal radioulnar (DRU) joint after distal radial fractures has a worse general outcome in patients before the usual age for development of osteoporosis. The hypothesis for this study was that patients with laxity also have less strength during supination and pronation. An apparatus for measuring strength in supination and pronation was constructed and validated. 20 patients were chosen from a cohort of 76 patients who had had distal radial fractures 6 years ago. The stability of the DRU-joint was tested manually by 2 examiners and the test's reliability was good. The DRU-joint was lax in 6 patients. No differences were found between patients with stable and lax DRU-joints as regards isometric grip strength, strength in supination or pronation or radiographic characteristics. However, subjectively evaluated hand function was impaired in those with lax DRU-joints. This was noted on the subjective part of the Gartland and Werley score and 13 questions concerning everyday activities. In conclusion, DRU-joint laxity was associated with impaired hand function, but no loss of strength in forearm rotation after distal radial fractures in non-osteoporotic patients. PMID- 12079012 TI - Mechanical evaluation of a carbonated apatite cement in the fixation of unstable intertrochanteric fractures. AB - We created three-part unstable intertrochanteric fractures in 6 pairs of aged, osteopenic, human, cadaveric femora. Fractures were reduced and fixed with a Dynamic Hip Screw (DHS) (Synthes, Paoli, PA). Two test groups were evaluated: 1. Fixation with DHS, and 2. Fixation with a DHS and calcium phosphate bone cement (Norian SRS (Skeletal Repair System)) augmentation of the fracture line and posteromedial calcar region of the proximal femur. Each femur was loaded to 1,650 N (2.5 body weight) for 10,000 cycles to simulate postoperative load transmission across the fracture construct during normal gait. The load was further increased successively by one body weight for another 10,000 cycles until failure. We evaluated fixation by measuring the amount of sliding of the lag screw of the DHS (shortening) and stiffness of the overall fracture construct (stability). SRS cement-augmented specimens had less shortening (1 mm versus 17 mm) and twice the initial construct stiffness compared to control specimens. PMID- 12079013 TI - Bacterial contamination of femoral head allografts from living donors. AB - We analyzed the bacterial contamination rate of femoral head allografts from living donors and determined the true bacterial load with cultures from the grafts in their entirety in a specially prepared medium. During 4 years we took swab cultures from 2,679 grafts of which 2,414 (90%) were negative. In a period of 12 months, grafts rejected for reasons other than infectious disease were cultured in their entirety to determine the true bacterial load. Of the 106 grafts included, 15 were contaminated. Microorganisms were isolated from 10 of the 91 initially swab culture-negative grafts (9%) and from only 5 of the 15 swab culture-positive grafts, most of which were normal skin contaminants. Swab culture negative grafts apparently may still be contaminated. It seems wise to subject all femoral head allografts from living donors to antibacterial processing. PMID- 12079014 TI - Overgrowth and correction of rotational deformity in 12 femoral shaft fractures in 3-6-year-old children treated with an external fixator. AB - We treated 11 young children (3-6 years old) who had uncomplicated femoral shaft fractures primarily with an external fixator. 9 children were available for follow-up and were evaluated for the amount of overgrowth and rotational deformity. All underwent a clinical examination and an MRI after mean 21 (13-25) months. The mean overgrowth was 0.4 (-0.3-1.1) cm and the anteversion angle showed a mean increase of 12 degrees, as compared to the contralateral femur. In 5 children with an anteversion angle difference of 10 degrees or more, a second MRI was done 4 years after the trauma. The mean anteversion angle difference of the femora in these 5 children had diminished from 15 degrees on the first MRI to 7.4 degrees on the second. 3 of the 5 children had a full correction of their rotational deformity. Growth did not correct the rotational deformity in the oldest child in this group. PMID- 12079015 TI - Combined intra-articular glucocorticoid, bupivacaine and morphine reduces pain and convalescence after diagnostic knee arthroscopy. AB - We studied the effect of intra-articullar saline vs. bupivacaine + morphine or bupivacaine morphine + methylprednisolone after diagnostic knee arthroscopy. In a double-blind randomized study, 60 patients undergoing diagnostic knee arthroscopy without a therapeutic procedure were allocated to groups receiving intra articular saline, intra-articular bupivacaine 150 mg + morphine 4 mg or the same dose of bupivacaine + morphine + intra-articular methylprednisolone 40 mg at the end of arthroscopy during general anesthesia. All patients were instructed to resume normal activities immediately after the procedure. Pain during movement and walking, leg muscle force and joint effusion, use of crutches and duration of sick leave were assessed. A combination of bupivacaine and morphine reduced pain, duration of immobilization and of convalescence. The addition of methylprednisolone further reduced pain, use of more analgesics, joint swelling and convalescence. PMID- 12079016 TI - Concomitant partial meniscectomy worsens outcome after arthroscopic anterior cruciate ligament reconstruction. AB - In this multi-center study involving 412 patients, we assessed the influence of concomitant partial meniscal resection on the medium-term clinical results after anterior cruciate ligament reconstruction. We performed a resection of minimum one-third of the medial or lateral menisci in 137 patients (group M) and found intact menisci in 275 patients (group NM). Those who had undergone previous meniscal surgery, subsequent meniscal surgery or a re-rupture of the anterior cruciate ligament graft during the follow-up were not included. After a median of 3 (2-6) years, the patients were reexamined by independent observers. Group M patients had more pain, swelling and laxity than those in group NM; they also had a worse classification according to the IKDC system, lower Lysholm scores and a greater proportion of patients with loss of motion. PMID- 12079017 TI - Arthrodesis by percutaneous fixation: patellofemoral arthrodesis in rabbits without debridement of the joint. AB - Arthrodesis is usually performed with joint resection or at least with removal of cartilage. Our recent experience with successful fusion in all 11 rheumatoid ankles treated only with percutaneous fixation questions the necessity of debridement of the joint before arthrodesis. In this rabbit study we tested the hypothesis that joints fuse because of rigid fixation. 9 skeletally mature loop ear dwarf female rabbits were operated on. With the knee in about 45 degrees of flexion, two 1.5 mm cortical screws were inserted through the patellofemoral joint in an anterior-posterior direction with the lag screw technique. 3 rabbits were excluded due to technical problems. Of the remaining 6 rabbits, 5 underwent bony fusion and 1 fibrous healing. Fusion was confirmed with Micro-CT in 2 cases and by histological examination in all 5 cases. In those 5 cases, bony fusion was seen in almost all areas with close contact. Therefore, fusion occurred not only in relation to the screws, but also between the screws and in the periphery of the patella. Our findings show that bony fusion can occur in a healthy joint without joint resection or debridement. PMID- 12079018 TI - The Weil osteotomy for treatment of dislocated lesser metatarsophalangeal joints: good outcome in 21 patients with 42 osteotomies. AB - Hardly any surgical methods are available for metatarsalgia caused by a dislocated lesser metatarsophalangeal joint (MTP) that do not sacrifice the joint. We reviewed retrospectively the outcome of 60 metatarsal Weil osteotomies for correction of dislocated lesser MTP joints in 31 patients. Between 1995 and 1996, 31 consecutive patients were treated with a Weil osteotomy at 2 institutions. The Weil osteotomy is an oblique osteotomy of the metatarsal neck and shaft, parallel to the ground surface, that controls shortening of the metatarsal by internal fixation with screws or pins. At an average final follow up of 30 (24-44) months, all patients were interviewed, using a standardized questionnaire based on the AOFAS Lesser Metatarsophalangeal-Interphalangeal Scale. Recurrent or transfer metatarsalgia, formation of callus, mobility and dislocation of the MTP were noted on physical examination. Dorsoplantar and lateral weightbearing radiographs taken preoperatively and at the time of final follow-up were examined for alignment of the metatarsal heads, subluxation or dislocation and for evidence of nonunion, or malunion of the metatarsal osteotomy. We had excellent results in 21 patients (42 osteotomies). A major complication was plantar penetrating hardware in 10 cases (3 screws and 7 pins). We conclude that the Weil osteotomy is a good method for correcting metatarsalgia caused by dislocation of the MTP joint. PMID- 12079019 TI - Hallux valgus in children: a 5-14-year follow-up study of 30 feet treated with a modified Mitchell osteotomy. AB - Between 1985 and 1995, 30 modified Mitchell osteotomies were performed in 18 children with hallux valgus, 12 bilateral and 6 unilateral. The mean age at surgery was 15 (10-18) years. The surgical modification consisted of diverging trapezoidal cuts, plantar displacement of the head, release of the lateral collateral ligament and the adductor insertion and Kirschner wire fixation of the osteotomy. At an average follow-up of 8 (5-14) years there were no nonunions, avascular necroses or recurrences. All the patients were satisfied with the cosmetic results, could use regular shoes and had no physical restrictions. Only 2 complained of occasional pain, thought to be secondary to transfer metatarsalgia. The presence of an open physis at the time of surgery did not affect the results. PMID- 12079020 TI - Release of netilmicin and vancomycin from cancellous bone. AB - First, we studied the effect of the following variables used for netilmicin- and vancomycin-impregnation of cancellous bone: a) antibiotic concentration of the impregnation fluid, b) time used for impregnation, c) pH of the impregnation fluid, d) the degree of bone morselizing and e) antibiotic combination. An increase in the antibiotic concentration of the impregnation fluid increased the amount of antibiotics released from bone. In addition, the amount of vancomycin eluted was also dependent on the time used for impregnation. The fraction of the total amount of netilmicin and vancomycin released after 24 h was 80% and 30%, respectively. More netilmicin and vancomycin were eluted from bone impregnated with antibiotics at pH 7 than the amount eluted from bone impregnated at pH 3. More netilmicin was eluted from fine morselized bone than from coarse morselized bone. By combining netilmicin and vancomycin in the impregnation fluid, the release of vancomycin was reduced. Secondly, we analyzed if the release of antibiotics from bone was complete: 99.9% of the total amount of netilmicin adsorbed to the bone was released by elution during 6 weeks. Finally, after implantation of netilmicin-impregnated bone in rabbit femur condyle, we measured netilmicin and vancomycin in serum: peak serum values of netilmicin were 4.2 (3.7 4.7) mg/L 2-3 h postoperatively. PMID- 12079022 TI - Treatment of aggressive fibromatosis: a retrospective study of 72 patients followed for 1-27 years. AB - We evaluated prognostic factors for local recurrence-free survival, including expression of estrogen receptors, after surgical treatment of aggressive fibromatosis in 72 patients (53 women) having primary tumors between 1970 and 1998. Their median age at diagnosis was 31(1 month-77 years) years. 50 patients had extraabdominal and 22 abdominal fibromatosis. Median tumor size was 4 (1-27) cm. 8 patients were treated with an intralesional resection, 32 with marginal, 31 with wide and with radical resection. They were followed for a median of 8 (1-27) years. The overall and local recurrence-free 5-year survival rates were 98% and 73%, respectively. Univariate analysis identified age, compartmentalization and tumor size as prognostic factors for local recurrence-free survival as well as radiotherapy in extraabdominal tumors. In the multivariate analysis, tumor size > 4 cm, extracompartmental location, inadequate margin and age < 32 years were independent negative prognostic factors for local recurrence. None of the tumors expressed estrogen receptors. In conclusion, aggressive fibromatosis as a high local recurrence rate, but a good prognosis, since almost no patients die of their tumor. PMID- 12079021 TI - Inhibition of polymethylmethacrylate particle-induced monocyte activation and IL 1beta and TNF-alpha expression by the antioxidant agent N-acetylcysteine. AB - We investigated the effectiveness of an antioxidant agent, N-acetylcysteine (NAC), in suppressing macrophage activation and mediator release in response to particulate debris. Polymethylmethacrylate (PMMA) particle-stimulated monocyte macrophages were cultured alone and with varying concentrations of NAC. Tumor necrosis factor alpha (TNFalpha) and interleukin-1beta (IL-1beta) expression in the resultant cultures were measured using enzyme-linked immunosorbant assays. The ultrastructural effect of treatment was also assessed by electron microscopy. Cell viability in the various cultures was measured to rule out an effect of cytotoxicity. NAC treatment reduced TNFalpha and IL-1beta expression by the monocyte-macrophages. Culturing with NAC was also associated with less ultrastructural activation of the monocytes. Furthermore, NAC was not associated with any adverse effect on cell viability in the concentrations used. Our findings demonstrate the effectiveness of the antioxidant N-acetylcysteine in suppressing the cell activation and TNFalpha release seen on exposure to wear debris. This represents a novel potential therapeutic method in the prevention or treatment of periprosthetic osteolysis. PMID- 12079023 TI - Infiltration of sarcomas into the hip joint: comparison of CT, MRI and histologic findings in 67 cases. AB - We analyzed the incidence, route, and characteristics of hip joint infiltration in pelvic or proximal femoral sarcomas. 67 patients with a sarcoma that originated around the hip joint (50 pelvic and 17 femoral) were included in this study. Preoperative CT and MRI were matched with the histological findings in tumor specimens. Tumor infiltration into the hip joint was suspected on the basis of preoperative imaging in 29 patients due to articular cartilage disruption, diffuse signal changes in the acetabulum or femoral neck, signs of a tumor in the joint, or markedjoint effusion. Of these 29 patients, 15 showed tumor invasion on histological examination. 12 of 31 chondrosarcomas, none of 12 Ewing's sarcomas, and 3 of 24 osteosarcomas infiltrated into the hip joint (p = 0.008). 10 of 26 low-grade sarcomas and 5 of 41 high-grade sarcomas infiltrated into the hip joint (p = 0.02). The joint infiltration rate of the chondrosarcomas was related to their size. Of 10 tumors originating in the acetabulum, 9 penetrated through or around the osseous-ligamentous junction and one through the acetabular cartilage. In 5 proximal femur lesions, all infiltrated the joint through the femoral neck, 3 of them also through the ligamentum teres. PMID- 12079026 TI - Traumatic bowing and Galeazzi fracture-dislocation--a report of 2 children. PMID- 12079025 TI - An unusual variant of fracture through the capitate bone--a case report. PMID- 12079024 TI - Musculoskeletal hydatid disease: a report of 13 cases. AB - This is a retrospective study of 13 patients with muscular hydatidosis--i.e., 4% of the 309 cases of hydatid disease treated in our department during 1983-1999. The commonest clinical finding was an asymptomatic and slowly growing mass (7). Puncture or incision of the mass was followed by an infection of the cystic cavity with fistulization in 2 patients. The immunological findings were false negative in 4 patients. MR images were obtained in 4 patients before diagnosis, and were highly suggetive of hydatid disease. The cystic cavities in all 9 patients subjected to radical surgery healed without chemotherapy. Radical surgery was not possible in 4 cases, in 3 of whom the sacrum was involved. Medical treatment of these patients did not eliminate the disease and new operations were necessary. PMID- 12079027 TI - Explaining the gender difference in self-reported suicide attempts: a nationally representative study of Norwegian adolescents. AB - Predictors of self-reported suicide attempts were studied cross-sectionally and prospectively in order to discern which variables may account for the gender difference in such reports. A representative sample of Norwegian students (N= 9,679) in grades 7-12 were followed from 1992 to 1994. More girls (10.4%) than boys (6.0%) reported a previous attempt and more girls (3.3%) than boys (1.9%) reported an attempt during the study period. The gender difference in previous suicide attempts was significantly reduced when depressed mood was controlled, and was no longer significant when disordered eating was included. There was no gender difference in future attempts when previous attempts, depressed mood, physical appearance, pubertal timing, and romantic involvement were controlled. Girls' higher level of risk factors may account for their higher level of self reported nonfatal suicidal behavior. PMID- 12079028 TI - Sexual assault history and suicidal behavior in a national sample of women. AB - This study examined demographic and psychosocial correlates of suicidal ideation and suicide attempts in women with histories of sexual assault in childhood and/or adulthood identified from a national sample of women. Multivariate analyses showed that women with histories of sexual assault in both childhood and adulthood reported significantly greater odds of lifetime suicide attempts, controlling for demographic factors and other psychosocial characteristics. As predicted, younger age (marginal), stressful life events (marginal), depression, PTSD, and alcohol dependence symptoms were also significantly associated with suicidal ideation. Furthermore, number of lifetime traumatic events and depression were each associated with lifetime odds of suicide attempts. Implications for studying the role of sexual trauma and other psychosocial factors in relation to women's suicidal behavior are discussed. PMID- 12079029 TI - Additive impact of childhood emotional, physical, and sexual abuse on suicide attempts among low-income African American women. AB - This study examines the association between exposures to multiple forms of childhood abuse (emotional, physical, sexual) and adult suicidal behavior in a sample of low-income, African American women. We hypothesized a linear relation between childhood abuse and risk for making a suicide attempt, such that the more forms of childhood abuse to which a woman was exposed, the greater her risk would be to make a suicide attempt. Logistic regression analyses revealed that, compared to women who did not report any experiences of childhood abuse, women who experienced one, two, or three forms of abuse were 1.83, 2.29, or 7.75 times more likely to attempt suicide, respectively (all statistically significant). Furthermore, compared to women who reported one or two types of abuse, women who reported all three types of abuse were statistically more likely to attempt suicide. These findings have implications for clinicians working with African American women who report either childhood abuse or exhibit suicidal behavior. PMID- 12079030 TI - Neighborhood predictors of hopelessness among adolescent suicide attempters: preliminary investigation. AB - The role of neighborhood factors in predicting hopelessness among adolescent suicide attempters was examined in this study. Forty-eight adolescents who attempted suicide were administered measures of hopelessness and depression. Family socioeconomic status (SES) was calculated based on family demographics and characteristics of neighborhood context were examined using a geocoding software package. Adolescents who lived in neighborhoods with weak social networks reported higher levels of hopelessness, even after controlling for SES and depression. These preliminary findings suggest that environmental context may play a role in the emotional status of adolescents who attempt suicide. PMID- 12079031 TI - Dialectical behavior therapy adapted for suicidal adolescents. AB - We report a quasi-experimental investigation of an adaptation of Dialectical Behavior Therapy (DBT) with a group of suicidal adolescents with borderline personality features. The DBT group (n = 29) received 12 weeks of twice weekly therapy consisting of individual therapy and a multifamily skills training group. The treatment as usual (TAU) group (n = 82) received 12 weeks of twice weekly supportive-psychodynamic individual therapy plus weekly family therapy. Despite more severe pre-treatment symptomatology in the DBT group, at post-treatment this group had significantly fewer psychiatric hospitalizations during treatment, and a significantly higher rate of treatment completion than the TAU group. There were no significant differences in the number of suicide attempts made during treatment. Examining pre-post change within the DBT group, there were significant reductions in suicidal ideation, general psychiatric symptoms, and symptoms of borderline personality. DBT appears to be a promising treatment for suicidal adolescents with borderline personality characteristics. PMID- 12079032 TI - Suicide survivors' perceptions of the treating clinician. AB - Seventy-one suicide survivors were surveyed about their perceptions of the clinicians who were treating their loved one at the time of death. Survivors provided information regarding their perceptions and attitudes toward clinician behaviors before and after the suicide and their perceptions of helpful and troubling aspects of clinician behaviors. Results indicated that survivors share a number of common opinions regarding the mental health care providers treating their loved ones. Several differences existed between survivors who consider lawsuits against mental health care providers versus those who do not. The implications of these findings for clinical practice, legal issues, surviving suicide, and future research are discussed. PMID- 12079033 TI - Situational determinants of inpatient self-harm. AB - Auto-aggressive individuals have a higher likelihood of engaging in interpersonal violence, and vice versa. It is unclear, however, whether ward circumstances are involved in determining whether aggression-prone patients will engage in auto aggressive or outwardly directed aggressive behavior. The current study focuses on the situational antecedents of self-harming behavior and outwardly directed aggression of psychiatric inpatients. Inwardly and outwardly aggressive behavior were monitored on a locked 20-bed psychiatric admissions ward for 3.5 years with the Staff Observation Aggression Scale-Revised (SOAS-R). A map of the ward was attached to each SOAS-R form, enabling staff members to specify locations of aggressive incidents. Time of onset, location, and provoking factors of auto aggressive incidents were compared to those connected to aggression against others or objects. Of a total of 774 aggressive incidents, 154 (20%) concerned auto-aggressive behavior. Auto-aggression was significantly more prevalent during the evening (i.e., 50% compared to 32%), and reached its highest level between 8 and 9 P.M. (17% compared to 7%). The majority of self-harming acts (66%) were performed on patients' bedrooms. Outwardly directed aggression was particularly common in the day-rooms (24%), the staff office (19%), the hallways of the ward (14%), and the dining rooms (10%). Provoking factors of auto-aggressive behavior are less often of an interactional nature compared to outwardly directed aggression. The results suggest that a lack of stimulation and interaction with others increases the risk of self-injurious behavior. Practical and testable measures to prevent self-harm are proposed. PMID- 12079034 TI - Is evaluative research on youth suicide programs theory-driven? The Canadian experience. AB - An in-depth review found that only 15 Canadian youth suicide programs had been evaluated over the last decades. Most of these were conducted in schools; general education on suicide was the most common strategy used. The descriptions of the programs were incomplete, and their theoretical bases never presented. All evaluations looked at program effects in accordance with the predominant experimental paradigm in evaluative research. Only two of the programs led to a reduction in suicidal behavior. Future evaluative research should place greater emphasis on the content of programs, especially their theoretical bases. PMID- 12079035 TI - Use of statistical techniques in studies of suicide seasonality, 1970 to 1997. AB - The effect of seasons on suicides has been suggested repeatedly. In order to reveal a true seasonal pattern, an appropriate statistical technique, which is sensitive to a specific type of cyclic variation in the data, must be chosen. This study is a review of the use of statistical techniques for seasonality and of some important characteristics of study samples that were evaluated from 46 original suicide seasonality articles published in major psychiatric journals. The results showed that statistical techniques were applied in a majority of articles, but they were commonly lacking regarding analyses, which compared seasonal patterns among subgroups of a population. In recent studies more sophisticated statistical techniques were utilized for seasonality, like spectral analyses, as compared with earlier studies, in which the emphasis was on chi square tests. Lack of reporting essential features of the data, such as the sample size and monthly values of suicides, were frequent. The calendar effect was adjusted only in 11 studies. Some recommendations concerning the methodological and reporting issues are summarized for future articles on the seasonal affect on suicides. PMID- 12079036 TI - Does legalized gambling elevate the risk of suicide? An analysis of U.S. counties and metropolitan areas. AB - This study addresses a possible link between suicide and casino gambling. Resident suicide rates are analyzed for (a) a 1990 cross-section of 148 U.S. metropolitan areas and (b) before and after the advent of legalized casinos in five U.S. counties. Data are drawn from government and gaming industry sources. In cross-section, metro area suicide is strongly correlated with region, accidental death and homicide rates, age and race composition, and economic vitality, followed by a modest net positive correlation with casino presence. By contrast, the time series analysis yields no evidence of a gambling effect. PMID- 12079037 TI - Selection strategies for limiting the increase in ascites while increasing growth in broilers. AB - The objective of the current study was to compare the changes in a fitness trait when selection is performed for 5, 10, and 20 generations on a production trait that influenced its expression. Responses to single-trait selection for growth based on phenotype or animal model predictions were compared by computer simulation. Two-trait index selection was performed when a trait, related to the fitness trait, was included in the index with the production trait. The phenotypic expression of the fitness trait among the sibs was also considered as a selection factor for single-trait and two-trait index selection. For a fixed increase in the expression of the fitness trait, mass selection produced a larger increase in the production trait than did use of standard animal model best linear unbiased prediction under single-trait selection. The reduction in the genotypic mean of the fitness trait was accompanied by an increase in its phenotypic expression. The use of sib information and an indicator trait reduced the level of expression reached by the fitness trait. PMID- 12079038 TI - Responses of two strains of female broiler breeders to a midcycle increase in photoperiod. AB - The influences of strain and a midcycle increase in photoperiod on the reproductive performance of broiler breeder hens were evaluated. At 22 wk of age, Strain X and Y pullets were randomly assigned to one of five groups [processed at 22 wk of age (A), processed after third oviposition (B), processed at 32 wk of age (C-32), processed at 40 wk of age (C40), and processed at 54 wk of age (C 54)] and were placed into individual laying cages. These groups were euthanized at different ages to analyze reproductive and carcass parameters. Photostimulation occurred at 22 wk of age when the photoperiod was increased from 8L:16D to 14L:10D. A light-tight wall partitioned the room into two halves. At 33 wk of age, half of the birds received an increase in photoperiod to 18L:6D, whereas the other half remained at 14L:10D. Body weight and abdominal fat pad weight increased with age. The number of large yellow follicles (LYF) decreased from 8.78 at sexual maturity to 4.60 at 54 wk of age. The number of atretic white follicles (WF) and small yellow follicles (SYF) also increased with age. Although there were no differences in BW, Strain Y had a greater egg weight throughout the laying cycle than Strain X. Peak hen-day egg production was 93.0%, and, overall, egg production was not affected by strain. Reproductive performance and ovarian morphology were not affected by the increase in photoperiod, presumably because peak egg production was so high. Carcass data demonstrated that frame size was determined by sexual maturity, whereas organ weights changed in response to reproductive status. PMID- 12079039 TI - The effects of feeding the direct-fed microbial, primalac, on growth parameters and egg production in single comb white leghorn hens. AB - Two commercial strains of Single Comb White Leghorn hens, HyLine W-36 and DeKalb XL, were reared separately as pullet flocks in an enclosed, environmentally controlled brood-grow house with three banks of quad-deck cages (310 cm2 per bird). There were 1,800 hens of each strain, which were divided into two groups, control feed and a direct-fed microbial (DFM) feed treatment. During the pullet rearing phase to 18 wk of age, the DeKalb birds had greater parameters of growth and overall feed cost than the HyLine birds. The overall feed cost of the DFM treatment was $0.02 per bird greater than controls. However, the DFM-treated feed improved the livability of the DeKalb birds by 2.68%. At 18 wk of age, 3,528 hens were transferred to an enclosed, mechanically ventilated layer house. The two strains were placed in two differently sized quad-deck layer cages with seven birds per cage; the cage sizes were 35.56 cm x 60.96 cm (2,267.73 cm2 or 310 cm2 per bird) and 14 cm x 32 cm (2,890.32 cm2 or 413 cm2 per bird). There were a total of 144 replications with each strain and DFM treatment combination made up of 36 replications equally divided between the high and low density cages. At 70 wk of age, hens receiving the DFM treatment exhibited greater mean egg weights (61.72 g) and percentages of extra large eggs (XLE, 52.06%) compared to that of controls (61.12 g and 48.98%, respectively). The treatment of 310 cm2/hen DFM produced the least (P < or = 0.05) large eggs; however, this treatment group produced the most (P < or = 0.05) XLE. The use of DFM resulted in a shift from smaller to larger eggs. Regardless of density, the DFM hens had lower (P < or = 0.05) feed cost (FC) compared to FC of the controls (a savings of $0.46/bird between the DFM and control in the 310 cm2 /hen treatment and a $0.83 savings/bird between the DFM and control in the 413 cm2/hen treatment). It was concluded that PrimaLac can improve egg size and lower feed costs, regardless of the bird density used in the present study. PMID- 12079040 TI - The effect of twenty-eight-hour ahemeral day lengths on carcass and reproductive characteristics of broiler breeder hens late in lay. AB - The effect of increasing the day length from 24 to 28 h on the reproductive and carcass characteristics of old broiler breeder hens was examined. A total of 128 birds aged 47 wk of age from three different strains; Classic, Experimental (EXP), and Feather Sexable Yield (FSY) were used. Selection was based on target BW and egg production rates; birds were housed in light-tight environment chamber facilities. Birds were feed restricted, and feed was allocated according to actual weekly BW relative to target BW. At 55 wk of age 125 broiler breeders were killed and dissected, and carcass and reproductive traits were examined. Overall, ahemeral day length negatively affected carcass and reproductive parameters. No significant differences were found for BW, yolk, albumen and shell weights, normal and atretic ovarian follicles, and breast, fat pad, and liver weights. Significantly lower production rates, higher amounts of defective eggs, lower oviduct and ovary weights, lower F1 follicle weight, poorer eggshell quality, and higher egg weights were found. Subsequently, broiler breeder hens increased egg weight at the expense of egg production when subjected to an ahemeral day length. It is not recommended that day length be increased this late in production because it results in reduced egg productivity. PMID- 12079041 TI - Influence of increased environmental complexity on leg condition, performance, and level of fearfulness in broilers. AB - We hypothesized that increased distance between resources and stimulation of foraging behavior, through altering the degree of environmental complexity by using moving lights and scattering whole wheat in the litter, would improve physical activity of broiler chickens. Increased activity may potentially improve leg condition and performance and decrease the level of fearfulness in broilers. To test this hypothesis, 1,800 1-d-old male broilers were divided into 40 groups of 45 birds each (10 birds/m2). Each group was assigned to one of four treatments (10 replicates) as follows: barrier treatment (B) contained three barriers placed between the drinker line and the feeder. The light treatment (L) consisted of brightly colored moving lights projected on the pen floor for four 1-h periods/d throughout rearing. For the wheat treatment (W) wheat was dispersed on the floor from Days 8 to 17. Control birds (C) were maintained under standard management. Body weights and consumption were obtained throughout the rearing period. Gait score (GS), tibia length and diameter, fluctuating asymmetry (FA), bone ash, tibial dyschondroplasia (TD), bone breaking strength, and tonic immobility (TI) were measured at slaughter age. Mortality, body weight, feed conversion, FA, bone ash, TD, bone breaking strength, and TI duration did not differ significantly among groups. L birds had a higher GS than C and B birds and W birds had a higher GS than C birds (P < 0.05). Provision of barriers significantly increased the diameter of the tibia diaphysis (P = 0.05), which is a promising result for further studies to improve leg condition. PMID- 12079042 TI - Broiler performance, body weight variance, feed and water intake, and carcass quality at different stocking densities. AB - The effects of four stocking and water nipple densities on broiler performance and carcass traits were measured in two trials. The stocking densities of 23.8, 17.9, 14.3, and 11.9 birds/m2 corresponded to 260, 195, 156, and 130 birds per pen, respectively. The water nipple densities were 5, 10, 15, and 20 birds per water nipple. Birds in Trial 1 were processed at Day 39 and those in Trial 2 were processed at Day 42. Water and feed were provided ad libitum and light was provided 23 h/ d. Water nipple density had no effect on broiler performance or carcass quality. Birds grown at 23.8 birds/m2 had lower BW (1,898 g) and carcass weights (1,334 g), whereas birds grown at 14.3 birds/m2 had the highest BW (1,985 g) and carcass weights (1,432 g). Although the treatment with 23.8 birds/m2 gave the lowest BW, the yield of broilers per unit of floor space was highest (46.0 kg/m2). The coefficient of variation for BW was higher in the treatment with 11.9 birds/m2 (15.3 %) than in the other treatments (13.0%). The birds in the treatment with 11.9 birds/m2 consumed the least feed (2,993 g/bird) and those in the 14.3 birds/m2 treatment consumed the most feed (3,183 g/bird). The amount of water consumed and the water to feed ratio was highest in the 23.8 birds/m2 treatment (5,546 mL/bird and 1.85 mL/g, respectively). Stocking density had no effect on mortality, breast yield, carcass grading, incidence of scratches, or carcass quality. It was concluded high yield per unit area with good carcass quality could be achieved when ventilation rate and air circulation were adequate. PMID- 12079044 TI - Recovery and utilization of useful by-products from egg processing wastewater by electrocoagulation. AB - The efficacy of a laboratory electrocoagulation (EC) system for treating egg processing plant waste-water (WW) is reported. For simulated and industrial egg processing WW, chemical oxygen demand, turbidity, and total suspended solids (TSS) were reduced 92 to 97%, 97%, and 99%, respectively, after treatment with EC. The final TSS concentration and turbidity values were 30 mg/L and 5 formazin turbidity units (FTU), respectively, similar to that of potable water standards. The recovered by-product solids had a similar pattern of essential amino acids compared to that of liquid whole egg and were comparable to the Food Agriculture Organization's essential amino acid profile for an ideal protein. The relative protein digestibilities of the recovered solids and a commercial corn meal averaged 130 and 56%, respectively, compared to liquid whole egg (set at 100%). An economic analysis of EC indicated that this treatment is economically feasible in that a savings of approximately $425,000 per year is possible in addition to recovering the capital equipment costs after about 14 mo of operation. These findings demonstrate that EC can be successfully applied to treat egg processing plant WW, yielding a high quality water suitable for recycling and valuable by products having a highly digestible protein and fat value. PMID- 12079043 TI - Carbohydrate-based cocktails that decrease the population of Salmonella and Campylobacter in the crop of broiler chickens subjected to feed withdrawal. AB - The efficacy of various carbohydrate-based cocktails in reducing the number of enteropathogens in the crops of broilers subjected to feed withdrawal was examined. Market-aged broilers that had been orally challenged with Salmonella typhimurium were provided the cocktails during a 12-h feed withdrawal. After feed withdrawal, the broilers were processed, and their crops were aseptically removed and weighed. Crops were then blended in distilled water, and the pH of the suspensions was measured electronically. Populations of S. typhimurium, Campylobacter, and lactic acid bacteria in the crop suspensions were enumerated. Findings indicated that significantly fewer S. typhimurium and Campylobacter were recovered from the crops of broilers that had been provided cocktails supplemented with sucrose than from the (Key words: crop, carbohydrate, feed crops of broilers provided cocktails supplemented with equal concentrations (wt/vol) of glucose. Furthermore, significantly fewer S. typhimurium were recovered from the crops of broilers provided cocktails supplemented with 2 to 10% sucrose than from the crops of broilers provided water or cocktails that were not supplemented with carbohydrates. The pH of the crop contents of broilers provided carbohydrate cocktails were lower than the pH of the crops of broilers provided water or cocktails that were not supplemented with carbohydrates. Consumption of the cocktails did not produce significant changes in the crop weights. Findings indicate that altering the composition of carbohydrate-based cocktails provided to broilers during feed withdrawal may affect the efficacy of cocktails in reducing the number of enteropathogens recovered from the crops of broilers. PMID- 12079045 TI - Dietary arginine intake alters avian leukocyte population distribution during infectious bronchitis challenge. AB - Although dietary arginine is a factor in immune function and disease resistance, the full range of effects has yet to be described. In this study, the effects of dietary arginine on leukocyte population changes were examined in the peripheral blood and the respiratory tract of chickens inoculated with infectious bronchitis virus (IBV) strain M41. At 2 wk of age, female line P2a White Leghorn-type chickens were randomly assigned to one of three diets with different arginine levels: a marginally deficient diet (0.5%), an adequate diet (1.0%), and a diet containing a high level of arginine (3.0%). All birds were inoculated with IBV at 4 wk of age, and then the peripheral blood and the respiratory lavage were collected at 1 and 7 d postinfection (DPI). The growth rate of birds that received 0.5% arginine was significantly lower than that of birds receiving 1.0 or 3.0% arginine, whereas the growth of the latter groups did not differ. The percentage and absolute number of heterophil (H) and the H/lymphocyte (L) ratio in the peripheral blood at 1 DPI significantly increased as dietary arginine increased. In the respiratory lavage at 1 DPI, the percentage of H also increased with dietary arginine increase. At 7 DPI, the percentage of CD8+ cells from birds fed the deficient diet was lower than those from birds fed the adequate diet and the diet containing a high level of arginine, whereas the cell surface density of CD8 antigen did not vary among groups. These results show that dietary arginine influences the character of the chicken cellular response to IBV and the distribution of responding leukocyte subpopulations in a target tissue for the infection. PMID- 12079046 TI - Expression of myostatin is not altered in lines of poultry exhibiting myofiber hyper- and hypoplasia. AB - Decades of selective breeding have yielded lines of poultry with substantial myofiber hyperplasia, vet little is known about what genes have been altered during the course of selection. Myostatin is a strong negative regulator of muscle mass in mice and cattle and could have been one of many genetic factors contributing to increased myofiber deposition in growth-selected lines of poultry. To test this hypothesis, the sequence and expression patterns of myostatin were analyzed in growth-selected lines of chickens and quail. The sequence of broiler myostatin cDNA, amplified via reverse transcription (RT)-PCR from embryonic muscle RNA, contained no missense mutations in the coding sequence when compared to that of White Leghorn layers, although two silent single nucleotide polymorphisms (SNP) were found. Northern analysis of myostatin transcripts from embryonic pectoralis and quadriceps showed no significant differences in expression levels between broiler and layer muscle RNA. However, levels of myostatin transcripts were greatly reduced in muscles of posthatch chicks compared to embryonic muscle. Myostatin protein was also present in broiler and layer embryonic muscle at similar levels. No significant polymorphisms or differences in RNA expression levels were found in embryonic muscles of divergently selected lines of Japanese quail. These results indicate that intense artificial selection in these growth-selected lines of poultry has neither silenced the expression of myostatin nor created null alleles via mutation in the lines analyzed. PMID- 12079047 TI - Collagen X expression in oviduct tissue during the different stages of the egg laying cycle. AB - The purpose of this investigation was to study the expression of type X collagen in the hen's oviduct. Type X collagen is a short-chain collagen that is present in the fibers of eggshell membranes, and there is evidence to suggest that it contributes to structural integrity. In situ hybridization and Northern blot analysis were used to study the expression of this important matrix constituent. The results demonstrated that gene expression was predominantly in the tubular gland cells of the isthmus segment of the oviduct. In contrast to observations with other matrix proteins, such as parathyroid hormone-related peptide and osteopontin, gene expression did not fluctuate with the position of the egg in the oviduct. PMID- 12079048 TI - Threonine requirement of commercial laying hens fed a corn-soybean meal diet. AB - Two experiments were conducted with Hy-Line W36 hens to evaluate their Thr requirement in a corn-soybean meal diet. Eight experimental diets were used in Experiment 1 with graded levels of Thr: 0.53,0.50, 0.48, 0.45, 0.42, 0.40, 0.37, and 0.35%. Seven experimental diets were used in Experiment 2 with graded levels of Thr: 0.58, 0.53, 0.50, 0.48, 0.45, 0.42, and 0.40%. Supplemental amino acids (AA) were added to all diets to ensure that Thr was the first-limiting AA. Egg production (EP), egg mass (EM), and daily Thr intake were significantly increased with increasing dietary Thr levels. Broken-line regression indicated a daily Thr requirement of 439.0 and 462.1 mg per hen per d in Experiment 1 and of 393.9 and 447.1 mg per hen per d in Experiment 2 for EP and EM, respectively. The Thr requirements to produce 1 g of EM were 8.76 and 9.44 mg in Experiments 1 and 2, respectively. PMID- 12079049 TI - The effects of toasting canola meal on body weight, feed conversion efficiency, and mortality in broiler chickens. AB - It is hypothesized that the moisture incorporated into canola meal (CM) during desolventization, as sparge steam, promotes toasting. Elimination of toasting of CM would result in higher digestible amino acid content, but it is not known if it is required to reduce anti-nutritional factors. Therefore, the objectives of this study were to determine if suspending the use of sparge steam would prevent toasting and to study the effects of toasting on broiler chicken performance. Conventional toasted CM (TCM) and a hexane laden, nontoasted CM (NTCM) were collected from a commercial crushing plant. NTCM was desolventized in a research desolventizer/toaster without the use of sparge steam, producing a yellow-colored meal. The meals were fed to broiler chickens from 0 to 39 d and replaced 0, 20, 40, 60, 80, and 100% of the soybean meal (SBM) in wheat-based diets. Elimination of toasting increased broiler weight from 0.606 and 2.148 to 0.618 and 2.181 kg at 19 and 39 d of age, respectively. The feed ratio (0 to 19 d) increased from 0.637 to 0.642, but toasting did not affect mortality. Total mortality and chronic heart failure between 19 and 39 d increased with level of CM addition from 5.2 to 13.9% and 1.9 to 9.6%, respectively. Chronic heart failure in males, but not females, was increased from 3.3 to 17.4% with increasing CM level. In conclusion, desolventization without sparge steam produced a nontoasted meal and improved broiler growth and feed efficiency in comparison to TCM. Therefore, NTCM could be fed to broiler chickens. PMID- 12079050 TI - Chia seed (Salvia hispanica L.) as an omega-3 fatty acid source for broilers: influence on fatty acid composition, cholesterol and fat content of white and dark meats, growth performance, and sensory characteristics. AB - Five thousand four hundred, 1-d-old, male, Ross 308, broiler chicks were fed for 49 d to compare diets containing 10 and 20% chia (Salvia hispanica L.) seed to a control diet. Cholesterol content, total fat content, and fatty acid composition of white and dark meats were determined at the end of the trial. A taste panel assessed meat flavor and preference. Cholesterol content was not significantly different among treatments; however, the 10% chia diet produced a lower fat content in the dark meat than did the control diet. Palmitic fatty acid content was less in both meat types when chia was fed, with differences being significant (P < 0.05), except for the white meat and the 20% chia diet. alpha-Linolenic fatty acid was significantly higher (P < 0.05) in the white and dark meats with the chia diets. Chia significantly lowered the saturated fatty acid content as well as the saturated:polyunsaturated fatty acid and omega-6:omega-3 ratios of the white and dark meats compared to the control diet. No significant differences in flavor or preference ratings were detected among diets. Body weight and feed conversion were significantly lower with the chia diets than with the control, with weight reductions up to 6.2% recorded with the 20% chia diet. PMID- 12079051 TI - Relative effectiveness of methionine hydroxy analog compared to DL-methionine in broiler chickens. AB - Two broiler experiments were conducted to assess the relative bioefficacy of liquid DL-Met hydroxy analog-free acid (MHA-FA) and DL-Met (DLM). Exponential regression analysis was used to determine biological efficacy based on body weight, feed conversion, and carcass responses to dietary Met source. In Trial 1, four graded inclusion levels of DLM and liquid MHA-FA (0.06, 0.12, 0.18, and 0.24%) were each added to a basal diet that met the nutrient and energy requirements of broiler chickens, with the exception of Met + Cys. In four additional treatments, diluted DLM (65%) was added at the same supplementation levels as pure DLM and liquid MHA-FA. In the 42-d trial, broilers responded significantly (P < 0.05) to the supplements. Regression analysis revealed that liquid MHA-FA was 68% (weight gain), 67% (feed conversion), 62% (carcass yield), and 64% (breast meat yield) as efficacious as pure DLM on an as-fed basis. Responses to liquid MHA-FA and diluted DLM were very similar at corresponding supplementation levels. Diluted DLM as an internal standard confirmed that exponential regression analysis was a statistically valid technique for determination of the relative efficacy of nutrient sources. In Trial 2, five graded inclusion levels of each DLM (0.040, 0.091, 0.152, 0.222, and 0.303%) and liquid MHA-FA (0.045, 0.102, 0.170, 0.250, and 0.350%) were added to a basal diet limiting in Met + Cys but adequate in all other nutrients and energy. Liquid MHA FA was 72% (weight gain), 51% (feed conversion), 48% (carcass yield), and 60% (breast yield) as efficacious as DLM on a weight-for-weight basis. PMID- 12079052 TI - Lipoprotein lipase mRNA expression in abdominal adipose tissue is little modified by age and nutritional state in broiler chickens. AB - Lipoprotein lipase (LPL)-catalyzed hydrolysis of plasma lipoproteins is a rate limiting step in the transport of lipids into the peripheral tissues of broiler chickens. The aim of the present study was to investigate whether LPL mRNA expression in adipose tissue is affected by age or nutritional treatments, with a view to reducing fat accumulation in broiler chickens. The study found that chicken LPL mRNA expression in abdominal adipose tissue did not differ significantly between chickens aged 4, 6, and 8 wk, but there was less expression of LPL mRNA in 2-wk-old chickens. In nutritional modulation, LPL mRNA levels in abdominal adipose tissues were not modified by 48-h feed deprivation or by subsequent refeeding for 48 h. In addition, expression of LPL mRNA was not significantly altered in chickens fed for 7 d on diets containing 8% olive oil (triolein rich), safflower oil (trilinolein rich), or linseed oil (trilinolenin rich). On the other hand, adipose LPL mRNA expression in chickens force-fed for 12 h with a trilinolenin (18:3) emulsion after 48-h feed deprivation was significantly decreased when compared to that in chickens force-fed with a triolein (18:1) or trilinolein (18:2) emulsion. Changes to LPL immunoreactive protein levels in chicken abdominal adipose tissues brought about by aging and nutritional manipulations were similar to those observed in relation to mRNA expression. These findings suggest that LPL mRNA expression in growing chickens is less responsive to aging and nutritional manipulation than in mammals, thereby indicating specificity of physiological response on broiler chicken LPL. PMID- 12079053 TI - Effect of fasting versus feeding oasis after hatching on nutrient utilization in chicks. AB - Two experiments were conducted to determine the effects of fasting, feeding Oasis, or feeding a corn-soybean meal (SBM) or dextrose-casein diet immediately after hatching on growth performance, MEn and apparent amino acid (AA) digestibility. In each experiment, five or six pens of eight chicks were assigned to one of five treatments: feeding a corn-SBM or dextrose-casein diet immediately after hatching, fasting for 48 h (no feed or water) after hatching, or feeding Oasis for 24 or 48 h (no water) after hatching. The fasting and feeding Oasis treatments were followed by feeding the corn-SBM diet for 21 d. Excreta were collected at 0 to 2, 3 to 4, 7, 14, and 21 d on feed for determination of MEn and AA digestibility. Fasting chicks for 48 h posthatching significantly depressed weight gains compared to all other treatments at 21 d of age. Although weight losses were not significantly different at 48 h posthatching, chicks fed Oasis for 48 h had significantly (P < 0. 05) higher weight gains at 1 and 3 wk of age than did chicks fasted for 48 h. Chicks fed the dextrose-casein diet had significantly (P < 0.05) higher weight gains at 48 h and 7 d posthatching than chicks fed the corn-SBM diet immediately after hatch in Experiment 2. As time on feed increased from 3 to 21 d, MEn increased substantially (25%) for all the treatments in which chicks were fed the corn-SBM diet. In addition, the MEn of the corn-SBM diet for chicks fed Oasis was significantly (P < 0.05) higher for than chicks fasted for 48 h. The MEn of the dextrose-casein diet was high immediately after hatching and increased only slightly (2%) with increasing age. Digestibility of most AA increased with increasing age for all the corn-SBM treatments, and the digestibility values were similar among treatments. However, AA digestibility values for the dextrose-casein diet were consistently high at all ages and much higher (P < 0.05) than the other dietary treatments. Our results indicated that feeding Oasis (compared to fasting) had a beneficial effect on growth performance and subsequent energy utilization of a corn-SBM diet and that Oasis may stimulate early gut development even though very early growth (0 to 48 h) is not affected. PMID- 12079054 TI - Embryonic development from first cleavage through seventy-two hours incubation in two strains of pekin duck (Anas platyrhynchos). AB - Embryonic mortality is a significant problem plaguing the commercial duck industry worldwide. Yet, an objective means to stage development of the duck embryo is lacking. Such a staging procedure, which is described in this study, is essential for the critical and reproducible assessment of embryo development. The morphological features associated with duck embryo development are very similar to those of the chicken, although the duck embryo develops more slowly. The staging scheme presented here provides objective morphological criteria describing the embryonic development of the duck. PMID- 12079055 TI - Ventilatory sensitivity to changes in inspired and arterial carbon dioxide partial pressures in the chicken. AB - To determine if intrapulmonary chemoreceptors could be the sole peripheral chemoreceptors responsible for ventilatory responses to inhaled CO2, we studied the relationships of minute ventilation, tidal volume, and respiratory frequency to inspired and arterial partial pressure of CO2 (P(I)CO2 and PaCO2) in decerebrate upright chickens when the birds inspired gases containing four low partial pressures of CO2 (0.02, 5.0, 8.2, and 11.4 torr). Because of variability in the measured variables from time to time in the same birds, as well as between birds, and because of the limited precision in measuring PaCO2, a 4 x 4 Latin square design and four statistical methods of data analyses (modified reduced major axis estimator, maximum likelihood estimator, average slope method, and summary slope method) were used. Tidal volume, minute ventilation, and PaCO2 increased, but respiratory frequency remained unchanged, as gases containing increased partial pressures of CO2 were inhaled. The results indicate that intrapulmonary chemoreceptors are not the sole receptors stimulated by inhaling gas containing even low partial pressures of CO2 and that stimulation of arterial and central chemoreceptors also occurs. Stimulation of these latter chemoreceptors may account for the increase in ventilation. Further, the results demonstrate the importance of maintaining a low level of environmental CO2 in poultry houses to minimize its influence on ventilation. PMID- 12079056 TI - Intravenous micro-particle injection and pulmonary hypertension in broiler chickens: cardio-pulmonary hemodynamic responses. AB - Experiments were conducted to determine whether intravenous injections of micro particles, having a size suitable to be trapped by the pulmonary precapillary arterioles, could be used to increase the pulmonary vascular resistance and thereby trigger an acute increase in the pulmonary arterial pressure (pulmonary hypertension). Anesthetized male broilers injected intravenously with inorganic (silica gel, polystyrene) or organic (cellulose, Sephadex) micro-particles developed an immediate pulmonary hypertension in proportion to the cumulative quantities of micro-particles injected. Micro-particle occlusion of a portion of the pulmonary arterioles forced the cardiac output to flow at a higher rate through the remaining vascular channels, thereby exposing a diffusion limitation characterized by undersaturation of the systemic arterial blood with oxygen (hypoxemia). The concurrent onset of systemic hypotension (reduced systemic arterial blood pressure) was not due to a reduction in cardiac output but rather was attributed to hypoxemic vasodilation of the systemic vasculature (reduced total peripheral resistance). Preliminary histological evaluations revealed micro particles lodged in inter- and intraparabronchial arterioles, surrounded by aggregates of thrombocytes and mononuclear leukocytes within 30 min post injection. These observations infer that intravenously injected micro-particles are carried to the lungs by the returning venous blood, where trapping of the micro-particles by the pulmonary vasculature triggers acute responses (increased pulmonary vascular resistance, pulmonary hypertension, systemic hypoxemia, systemic hypotension) that mirror those previously observed following acute occlusion of one pulmonary artery. Additional studies will be required to determine the extent to which the focal immune response to trapped micro particles promotes local vasoconstriction that amplifies the pulmonary hypertension attributable to direct physical obstruction of precapillary arterioles. PMID- 12079057 TI - Intermittent biotelemetric monitoring of electrocardiograms and temperature in male broilers at risk for sudden death syndrome. AB - Biotelemetry was used to acquire electrocardiograms (ECG) and temperature measurements in a study of male broilers at risk for sudden death syndrome (SDS), a fatal condition that may have underlying cardiovascular mechanisms. Day-old (Day 1) Arbor Acres x Arbor Acres male chicks were randomly assigned to two different diets: control (Diet A) and one that contained elements that contribute to SDS (Diet B). The heaviest birds in each group on Day 13 underwent surgery on Day 15 to have transmitters with temperature sensors and ECG electrodes implanted. After surgery, three controls and three implanted birds from each diet group were kept in individual cages and exposed to 23 h of light and 1 h of darkness during each 24-h cycle. Implantation did not affect weight gain between Days 13 and 22 (P = 0.396). Temperature measurements and 1-min ECG were taken every 15 min. Heart rate and heart rate variability were measured from three 2-s segments in two dark and two light period samples during Days 17 to 19. Diet B decreased weight gain (P = 0.045), lowered heart rate (P < 0.0001), and increased internal temperature (P < 0.0001). Heart rate variability was lower during dark versus light periods (P = 0.004), which indicates that the birds rested during the dark periods, but was not affected by diet (P = 0.651). Thus, biotelemetry provided a useful method for intermittent physiological monitoring of poultry on different diets and under changing environmental conditions. PMID- 12079058 TI - Use of ultrasonography to characterize ovarian status in chicken. AB - Much research has been conducted to investigate the effects of environmental and nutritional treatments on ovarian development in poultry. However, to investigate the ovary, the hen must be killed, and thus, lifelong egg production can only be inferred. To date, the ability to noninvasively determine ovarian status has not been available. Improvements in ultrasound technology now make it possible to observe ovarian condition in vivo, thereby allowing for repeated sampling of the same bird over an entire egg production cycle. In the current study, large yellow follicles (LYF; diameter greater than 10 mm) were characterized in broiler breeder hens using Aloka ultrasound diagnostic equipment. Ultrasound images were used to determine the number and diameter of the LYF as well as the presence of an egg in the oviduct. Immediately following ultrasonography, hens were killed and dissected to determine the number and diameter of LYF. From the ultrasound images, the number of LYF +/- 1 was predicted with 96.3% accuracy in Experiment 1 and 93.3% accuracy in Experiment 2. Diameter measurements were used to classify follicles hierarchically. Of the birds determined via dissection to have multiple hierarchies, 77.8% were identified with ultrasound. All regressing or regressed ovaries were correctly identified with ultrasound. PMID- 12079059 TI - Effect of electron beam irradiation on poultry meat safety and quality. AB - The purpose of this study was to determine the effectiveness of electron beam irradiation at doses of 1.0 and 1.8 kGy on the elimination of bacteria from boneless, skinless chicken breasts without significantly altering product quality. Microbial testing was conducted in triplicate using a whole carcass rinse method with each nonirradiated control group and an irradiation treatment group consisting of 10 samples. Results indicated that mean counts for coliforms, generic Escherichia coli, and psychrotrophs were 3.13, 3.26, and 1.92 log10 cfu/200 mL rinsate, respectively, in the control samples. However, these populations were not detected after the samples were irradiated with 1.0 or 1.8 kGy. Mean count of 4.60 log10 cfu/200 mL rinsate was detected for aerobic bacteria in the control samples. Irradiation doses of 1.0 and 1.8 kGy reduced the levels to 2.23 and 1.62 log10 cfu/200 mL rinsate, respectively. Irradiation also rendered the fillets free of Salmonella and Campylobacter. Consumer taste panels (product stored for 0, 14, and 28 d at 0 C) indicated that, at Day 0, there were no differences among controls and treatment groups for any of the quality attributes tested. At Day 14, texture and flavor attributes were lower for the irradiated groups. At Day 28, samples irradiated with 1.0 and 1.8 kGy were less desirable with decreased texture, flavor, and overall acceptability. Degree of lipid oxidation also increased as storage time and level of irradiation increased. Irradiated samples also had higher a* values, indicating they were pinker in color. PMID- 12079060 TI - Evaluation of chemical disinfectants for the elimination of Salmonella biofilms from poultry transport containers. AB - Containers used in transporting live poultry between production and processing units are a primary source of contamination for processed poultry products. Because disinfection of transport containers (TC) has been difficult to accomplish, it is probable that the choice of appropriate disinfectant and its application are partially or wholly responsible for the failure to adequately eliminate pathogens from TC. Therefore, 13 commercial disinfectants were selected and evaluated for their capacities to destroy Salmonella. The disinfectants were applied in various concentrations on prescribed areas (10 cm diameter circle) of galvanized steel surfaces (representative of TC material) that were artificially contaminated with Salmonella (10(8) cfu/mL) with a mixture of organic material. Likewise, coupons (1.9 cm2) made of the same metallic surfaces and covered with biofilms of Salmonella spp. were tested in the same manner for each disinfectant. Two of the disinfectants completely eliminated Salmonella on the artificially contaminated and biofilm-covered surfaces. These compounds produced logarithmic reductions in Salmonella populations as high as 7.18 within 2 min. One of the two effective disinfectants contained sodium hypochlorite and was effective at a concentration of 0.05% (vol/vol). The other disinfectant was an alkaline peroxide compound and was effective at a concentration of 1% (wt/vol). Evaluation of these two disinfectants under simulated conditions suggested that application under the prescribed regimen could result in effective elimination of Salmonella from TC within a limited period. PMID- 12079061 TI - Influence of selection for breast muscle mass on myosin isoform composition and metabolism of deep pectoralis muscles of male and female turkeys. AB - Advances in genetic selection and nutrition have resulted in rapid growth rates and increased muscle mass, predisposing turkeys to muscle disorders such as deep pectoral myopathies and increasing the incidence of pale, soft, and exudative muscle. The objective of this study was to determine if selection for breast muscle mass created an increase in anaerobic capacity of the deep pectoralis muscle. A total of 67, 18-wk-old, male and female turkeys from two male (tom) lines and one female (hen) line were used. Each bird was anesthetized and one deep pectoralis muscle was electrically stimulated via the pectoral nerve. Muscle pH was recorded every 30 s for 4 min of stimulation and every 1 min for a 10-min recovery period. Non-stimulated muscles, contralateral to the stimulated side, were assayed for lactate dehydrogenase (LDH) and glyceraldehyde phosphate dehydrogenase (GAPDH). Myosin isoforms were resolved with SDS-PAGE. Line or gender had no effect on rate of pH decline during or after stimulation. Declines in pH during stimulation were greater than during the recovery period (0.06 vs. 0.02 U/min). The lightweight male line (LM) had the greatest breast muscle mass as a percentage of body weight (P < 0.05) and the greatest LDH [293 mmol nicotinamide adenine dinucleotide (NADH) min(-1)microg(-1); P < 0.0001] and GAPDH (0.4452 mmol NADH min(-1)microg(-1); P < 0.05) activities. Hens had greater percentages breast weight than males (P < 0.05) and a tendency for increased enzyme activities. The LM line had the largest ratio (2.33:1) (P < 0.05) of adult to-neonatal myosin. Genetic selection for breast muscle mass resulted in an increased ratio of adult-to-neonatal myosin and increased anaerobic capacity. This effect on myosin isoform composition and anaerobic capacity supports handling modifications that are line specific to minimize meat quality defects. PMID- 12079062 TI - Trichloroacetic acid in the environment. AB - Suppositions that the trichloroacetic acid (TCA, CCl3C(O)OH) found in nature was a consequence solely of the use of chlorinated hydrocarbon solvents prompted this critical review of the literature on its environmental fluxes and occurrences. TCA is widely distributed in forest soils (where it was rarely used as an herbicide) and measurements suggest a soil flux of 160 000 tonnes yr(-1) in European forests alone. TCA is also produced during oxidative water treatment and the global flux could amount to 55 000 tonnes yr(-1) (from pulp and paper manufacture, potable water and cooling water treatments). By contrast, the yields of TCA from chlorinated hydrocarbon solvents are small: from tetrachloroethene 13 600 tonnes yr(-1) and from 1,1,1-trichloroethane 4300 tonnes yr(-1) on a global basis, at the atmospheric burdens and removal rates typical of the late 1990s. TCA is ubiquitous in rainwater and snow. Its concentrations are highly variable and the variations cannot be connected with location or date. However, there is no significant difference between the concentrations found in Chile and in eastern Canada (by the same analysts), or between Malawi and western Canada, or between Antarctica and Switzerland, nor any significant difference globally between the concentrations in cloud, rain and snow (although local enhancement in fog water has been shown). TCA is present in old ice and firn. At the deepest levels, the firn was deposited early in the 19th century, well before the possibility of contamination by industrial production of reactive chlorine, implying a non-industrial background. This proposition is supported by plume measurements from pulp mills in Finland. TCA is ubiquitous in soils; concentrations are very variable but there are some indications that soils under coniferous trees contain higher amounts. The concentrations of TCA found in plant tissue are region-specific and may also be plant-specific, to the extent that conifers seem to contain more than other species. TCA is removed from the environment naturally. There is abundant evidence that soil microorganisms dehalogenate TCA and it is lost from within spruce needles with a half-life of 10 days. There is also recent evidence of an abiotic aqueous decarboxylation mechanism with a half-life of 22 days. The supposedly widespread effects of TCA in conifer needles are not shown in controlled experiments. At concentrations in the needles of Scots pine similar to those observed in needles in forest trees, changes consequent on TCA treatment of field laboratory specimens were almost all insignificant. PMID- 12079063 TI - Behavior of dioxin during thermal remediation in the zone combustion process. AB - In the previous study, a new process concept for the thermal remediation of particulate/powder materials contaminated by polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) has been verified. It leads to removal efficiencies of more than 99.9% in the soil contaminated by PCDD/Fs in terms of toxicity equivalent quantity (TEQ). However, details of the reactions and phenomena during the process, i.e., decomposition, vaporization, reformation and trap of PCDD/Fs and their relating compounds, have not sufficiently been clarified yet. The present study aims to examine experimentally the transport and fate of PCDD/Fs in the process. In the experiment, a laboratory-scale process simulator and a soil sample preliminary mixed with octachlorinated dibenzo-p-dioxin spiked by carbon 13 isotope (13C-OCDD) were used. The distribution of 13C-OCDD in the soil bed during the process was measured by applying a quench technique that rapidly cools down the bed. Further, the total amount of 13C-OCDD discharged with outlet gas was measured. Using the obtained data, mass balance of 13C-OCDD in the process was estimated. The results show that about 99% of 13C-OCDD preliminary admixed with the soil was decomposed rather than released to the outlet gas. Only a trace amount of 13C-OCDD remained in the treated soil. In addition, a very small amount of other congeners having the 13C-cycles was detected in the treated soil and outlet gas although its TEQ values are not significant. These were probably formed by dechlorination reactions occurring in the process. PMID- 12079064 TI - A microcosm test for potential mineralization of chlorinated compounds under coupled aerobic/anaerobic conditions. AB - In this study, the feasibility of using a mineralization test under coupled aerobic/anaerobic conditions was demonstrated. The coupling of anaerobic methanogenic and aerobic methanotrophic conditions in a microcosm required the presence of both a carbon source for anaerobic metabolism and oxygen for aerobic metabolism. These requirements were fulfilled by using a slow hydrolyzing organic matter along with intermittent addition of oxygen to the bottle headspace. Perchloroethylene (PCE) mineralization tests confirmed the effectiveness of the proposed methodology as well as PCE mineralization under coupled conditions. PMID- 12079065 TI - Effect of organic acids on adsorption and desorption of rare earth elements. AB - Effect of citric, malic, tartaric and acetic acids on adsorption of La, Ce, Pr and Nd by and desorption from four typical Chinese soils was studied. Generally, adsorption capacities of rare earth elements (REEs) were significantly correlated with the cation exchange capacity (CEC) of soils. In the presence of acetic acids adsorption of REEs was similar to that in the presence of Ca(NO3)2. However, in the presence of citric, malic and tartaric acids adsorption of REEs by Heilongjiang, Zhejiang and Guangdong soils decreased to varying extents if compared with that in the presence of nitrate and acetic acid. The significance of suppression followed the order of citric acid > malic acid > tartaric acid > acetic acid, which was consistent with the order of stability of complexes of REEs with these organic acids. However, the adsorption increased with increasing equilibrium solution pH. For Jiangxi soil with low soil pH, CEC and organic matter these organic acids exerted an even more serious suppression effect on the adsorption of REEs. Another feature of the relationship between the adsorption of REEs and equilibrium solution pH was that the adsorption of REEs decreased with increase of pH from 2 to 4.5 and then slightly increased with further increase of pH. Desorption of REEs varied with soils and with organic acids as well. REEs were released easily from Heilongjiang, Zhejiang and Guangdong soils in the presence of organic acid. Generally, desorption of REEs decreased with increasing equilibrium solution pH. Effect of organic acids on desorption of REEs from Jiangxi soil was more complicated. In the presence of citric and malic acids no decrement and/or slight increase in desorption of REEs were observed over the equilibrium solution pH from 3 to 6.5. The reasons for this were ascribed to the strong complexing capacity of citric and malic acids and low soil pH, CEC and organic matter of Jiangxi soil. PMID- 12079066 TI - Comparison of modified montmorillonite adsorbents. Part I: Preparation, characterization and phenol adsorption. AB - This study concerns with the development of modified montmorillonites as adsorbents for water treatment. Polymeric aluminium and iron intercalated forms of montmorillonites have been prepared in the absence and presence of an alkylammonium cationic surfactant (Hexdecyl-trimethyl-ammonium bromide, HDTMA). Montmorillonites intercalated with polymeric Al, Fe, Fe/Al (2:1 Fe to Al ratio in solution), possess large N2 Brunauer-Emmett-Teller (BET) surface areas. XRD data also shows trace amounts of illite and plagioclase within the clay materials. Montmorillonites intercalated with HDTMA, polymeric Fe/HDTMA, polymeric Al/HDTMA and polymeric Fe/Al/HDTMA (1:1 metal to surfactant molar ratio in solution) undergo some losses of N2 BET surface areas. Preliminary adsorption studies on phenol have shown that polymeric Al/HDTMA- and HDTMA-only-modified montmorillonites possess a good affinity for phenol, whereas the polymeric Al/Fe modified- and starting montmorillonites have little affinity for phenol adsorption. PMID- 12079067 TI - Bioassay-directed chemical analysis of River Elbe surface water including large volume extractions and high performance fractionation. AB - A bioassay-directed fractionation and identification (toxicity identification evaluation procedure) was performed on extracts of 10 1 River Elbe water samples. The experimental method included a SDB-1 solid phase extraction followed by RP HPLC fractionation and subfractionation. Chemical analysis by GC-MS as well as acute toxicity testing using a luminescent bacteria assay were conducted in the respective fractions. Many substances were identified, among which were pesticides and pharmaceuticals, but many compounds remained unknown. PMID- 12079068 TI - Dissolution rates of three high explosive compounds: TNT, RDX, and HMX. AB - Incidental exposure to high explosive compounds can cause subtle health effects to which a population could be more susceptible than injury by detonation. Proper source characterization is a key requirement in the conduct of risk assessments. For nonvolatile solid explosives, dissolution is one of the primary mechanisms that controls fate and transport, resulting in exposure to these compounds remote from their source. To date, information describing dissolution rates of high explosives has been sparse. The objective of this study was to determine the dissolution rates of three high explosive compounds, 2,4,6-trinitrotoluene (TNT), hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), and octahydro-1,3,5,7-tetranitro 1,3,5,7-tetrazocine (HMX), in dilute aqueous solutions as a function of temperature, surface area, and energy input. To determine each variable's impact on dissolution rate, experiments were performed where one variable was changed while the other two were held constant. TNT demonstrated the fastest dissolution rate followed by HMX and then RDX. Dissolution rate correlation equations were developed for each explosive compound incorporating the three aforementioned variables, independently, and collectively in one correlation equation. PMID- 12079070 TI - The effect of dissolved glyphosate upon the sorption of copper by three selected soils. AB - The effect of the pesticide glyphosate (GPS) on adsorption processes of copper onto three soils of different characteristics has been studied. Cu adsorption decreases in general with increasing GPS concentration in solution, due principally to the lower equilibrium pHs, although this is not the only variable affecting copper adsorption. For the same pH values, Cu adsorption is higher in two of the three soils in the presence of GPS, but for the third soil, Cu adsorption is higher in the absence of GPS. This behavior is explained by the possibility of GPS adsorption on these soils and by the formation of Cu-GPS complexes in solution. The soils showing a higher Cu adsorption in the presence of GPS than in its absence for the same pH are able to adsorb this pesticide. In these soils, copper can be adsorbed directly on the soil surfaces, and also through the formation of bonds with GPS previously adsorbed. The third soil was not able to adsorb GPS. Consequently, all the pesticide remained in solution, forming strong Cu complexes with low tendency to be adsorbed on this soil. For this reason, the concentration of free Cu in solution is drastically reduced, and the adsorption of copper on this soil is lower. PMID- 12079069 TI - Evidence for destruction of PCBs by the OH radical in urban atmospheres. AB - Evidence for reaction of polychlorinated biphenyl (PCB) congeners with the hydroxyl (OH) radical in the troposphere was observed in diurnal variations in ambient gas-phase PCB concentrations at three urban sampling sites located in the Chicago, IL; Baltimore, MD; and Jersey City, NJ urban/industrial areas. The magnitude of the depletion of individual PCB congeners decreased by about 10-20% for each additional chlorine substituent, reflecting slower reaction rates for higher MW congeners with the OH radical. Octa- and nonachlorobiphenyls, which are largely unreactive with the OH radical, were used as tracers to investigate the effects of dilution on diurnal variation. The environmental rate constants for disappearance of the PCBs range from about 1.0 day(-1) for trichlorobiphenyls to about 0.3 day(-1) for hexachlorobiphenyls. Assuming a OH radical concentration of 3 x 10(6) molecules cm (-3), the second-order rate constants for reaction of specific congeners with the OH radical are consistent with laboratory measurements. More importantly, the relative reactivity of PCB homologues agrees well with the relationship predicted by other researchers from laboratory measurements, suggesting that losses of PCBs during daytime tropospheric transport are due at least in part to reactions with the OH radical. PMID- 12079071 TI - Atmospheric deposition of metallic pollutants over the Ligurian Sea: labile and residual inputs. AB - Atmospheric fluxes of six trace metals (Cd, Cr, Cu, Ni, Pb and Zn) with Al as a crustal reference were measured at Cap Ferrat (French Riviera) between February 1997 and July 1998. An original sampling protocol enabled the separation of labile (seawater at pH 2) and residual fractions in the total atmospheric input. Median acid-labile fractions were 91%, 69%, 83%, 84%, 97% and 98% of the total for Cd, Cr, Cu, Ni, Pb and Zn, respectively. Under the conditions used, lability of individual metals is related to the anthropogenic component of the samples. Enrichment factors and anthropogenic fraction are estimated for each metal. Some interannual changes are investigated (Pb, Zn). The observed increase of Zn inputs may be linked to local input from the Nice district waste plant (commissioned in 1988), 6.5 km away. PMID- 12079072 TI - Heavy metal extractable forms in sludge from wastewater treatment plants. AB - The analysis of heavy metals is a very important task to assess the potential environmental and health risk associated with the sludge coming from wastewater treatment plants (WWTPs). However, it is widely accepted that the determination of total elements does not give an accurate estimation of the potential environmental impact. So, it is necessary to apply sequential extraction techniques to obtain a suitable information about their bioavailability or toxicity. In this paper, a sequential extraction scheme according to the BCR's guidelines was applied to sludge samples collected from each sludge treatment step of five municipal activated sludge plants. Al. Cd, Co, Cu, Cr, Fe, Mn, Hg, Mo, Ni, Pb, Ti and Zn were determined in the sludge extracts by inductively coupled plasma atomic emission spectrometry. In relation to current international legislation for the use of sludge for agricultural purposes none of metal concentrations exceeded maximum permitted levels. In most of the metal elements under considerations, results showed a clear rise along the sludge treatment in the proportion of two less-available fractions (oxidizable metal and residual metal). PMID- 12079073 TI - Evaluation of biological integrity of a small urban stream system by investigating longitudinal variability of the fish assemblage. AB - We investigated the integrity of fish assemblages of a small anthropogenically impacted urban stream system in south-western Germany. Heavy point and non-point pollution physiologically stresses the aquatic fauna while migration barriers and river morphological alterations negatively influence habitat availability. Investigations conducted in 1998 and 1999 revealed that the fish assemblage was altered and several site-specific fish species were missing while non-site specific species were relatively common. Highest species diversity was recorded in a reach downstream of the main migration barrier, while upstream only a few species were present. In general, a shift to ubiquitous and limnophilic species as well as to less pollution sensitive species was observed. Adult fish dominated the population structure throughout the stream course. Recruitment was confined mainly to the less polluted headwater. In the present study we introduce an assessment system based on the similarity of the potential natural and the actually established fish assemblage as demanded by the Water Framework Directive of the European Union for the evaluation of the ecological status of a fish assemblage in a stream [EC of the European Parliament and of the Council of Europe establishing a framework for community action in the field of water policy. Brussels, 30 June, 2000. 1997/0067 (COD)]. Both, stream morphological quality and pollution data sampled within the two study years confirm the validity of the suggested assessment method. PMID- 12079074 TI - Comparative environmental assessment of biocides used in antifouling paints. AB - In response to increasing scientific evidence on the toxicity and persistence of organotin residues from antifouling paints in the aquatic environment, the use of triorganotin antifouling products was banned on boats of less than 25 m length in many countries during 1987. Alternatives to tributyltin (TBT) paint are mainly copper based coatings containing organic booster biocides to improve the efficacy of the formulation, and have been utilised on small boats for the last 10 years. With policies encouraging a total ban on TBT, it is expected that these biocides will be used to a greater extent in the future. Limited data and information are available on the environmental occurrence, fate, toxicity, and persistence of these biocides, and thus any decisions on policies regulating antifoulants cannot be fully informed. In this study, a multicriteria comparison of alternative biocides, based on a general assessment of available information in the literature, provided support for the use of the precautionary principle with respect to policies on antifouling products. This assessment was validated by a more detailed comparison of four selected biocides and TBT. Results indicate that TCMS pyridine and TCMTB demonstrate environmental characteristics similar to TBT and thus detail risk assessments are needed before their use is permitted. The widespread use of the other biocides should be allowed only after research to fill the gaps in knowledge with respect to their toxicity and persistence in aquatic environments. PMID- 12079075 TI - Pathway of CH4 formation in anoxic rice field soil and rice roots determined by 13C-stable isotope fractionation. AB - In anoxic rice fields methane is produced by either reduction of CO2 or cleavage of acetate. We measured the delta 13C-values of CH4 and CO2, acetate and organic carbon during time course experiments with anoxic methanogenic soil and root samples and used these values to calculate the fractions of CH4 (and acetate) produced from CO2 reduction. Comparison with radiotracer and/or inhibitor studies constrained the kinetic fractionation factors used for calculation. The fractionation factors for the conversion of CO2 to CH4 and of acetate to CH4 were on the order of alpha = 1.07 (epsilon = -70%) and epsilon > or = - 20%, respectively. The pathway of CH4 production changed with time of anoxic incubation. Anoxic slurries of rice field soil first produced CH4 predominantly (>50%) from CO2, then predominantly (>80%) from acetate and finally (after about one month) according to the theoretically expected ratio (33% CO2 and 67% acetate). Anoxic rice roots, on the other hand, initially produced CH4 exclusively from CO2, followed by contribution of acetate of about 40-60%. Rice roots also produced acetate that partially originated (< or = 1 30%) from reduction of CO2 as determined by calculation of isotopic fractionation using fractionation factors from the literature. The results demonstrate that there is quite some variability in pathways of CH4 production, and also indicate that isotopic fractionation factors may be different in different habitats and change with time. PMID- 12079076 TI - Atmospheric nitrous oxide: patterns of global change during recent decades and centuries. AB - Data from weekly global measurements of nitrous oxide from 1981 to the end of 1996 are presented. The results show that there is more N2O in the northern hemisphere by about 0.7 +/- 0.04 ppbv, and the Arctic to Antarctic difference is about 1.2 +/- 0.1 ppbv. Concentrations at locations influenced by continental air are higher than at marine sites, showing the existence of large land-based emissions. For the period studied, N2O increased at an average rate of about 0.6 ppbv/year (approximately 0.2%/year) although there were periods when the rates were substantially different. Using ice core data, a record of N2O can be put together that goes back about 1000 years. It shows pre-industrial levels of about 287 +/- 1 ppbv and that concentrations have now risen by about 27 ppbv or 9.4% over the last century. The ice core data show that N2O started increasing only during the 20th century. The data presented here represent a comprehensive view of the present global distribution of N20 and its historical and recent trends. PMID- 12079077 TI - Correction for water vapor in the measurement of atmospheric trace gases. AB - The presence of water vapor in a sample of air reduces the concentration of a trace gas measured from the sample. We present a methodology to correct for this effect for those cases when the concentration of the trace gas has already been measured from a wet sample. The conversion or correction factor that takes the wet mole fraction to a dry mole fraction is determined by the mixing ratio of water vapor inside the sampling canister. For those samples where the water vapor is saturated inside the canister, the water vapor mixing ratio is largely determined by laboratory conditions; for the unsaturated samples, the mixing ratio is determined by station conditions. If the meteorology at the sampling station is known, the equations presented here can be used directly to calculate the appropriate correction factor. For convenience, we use climatological data to derive average monthly correction factors for seven common global sampling sites: Barrow, AK, US (71 degrees N, 157degrees W); Cape Meares, OR, US (45 degrees N, 124 degrees W); Mauna Loa, HI, US (19 degrees N, 155 degrees W); Ragged Point, Barbados (13 degrees N, 59 degrees W); American Samoa (14 degrees S, 171 degrees W); Cape Grim, Tasmania, Australia (41 degrees S, 145 degrees E); South Pole (90 degrees S). These factors adjust wet mole fractions upwards within a range of 0.002% for the South Pole to over 0.8% for saturated sites. We apply the correction factors to wet nitrous oxide (N2O) mole fractions. The corrected data are more consistent with our understanding of N2O sources. PMID- 12079078 TI - Soil CO2 fluxes in cereal land use of the Spanish plateau: influence of conventional and reduced tillage practices. AB - From November 1998 to October 2000, measurements of soil respiration were performed on the Spanish plateau for two patches of non-irrigated barley, one managed with conventional tillage (CT) and the other with reduced tillage (RT). Soil CO2 flux showed seasonal variation on both patches, with an increase from March to October, peaking in May, and a decrease during the winter period by a factor of around 2. The mean value for both combined years was 2.03 and 1.70 micromol m(-2) S(-1), in the CT and RT patches, respectively. In order to analyse the influence of RT on soil CO2 flux, two tests were performed. The first one was the Kruskal-Wallis test to compare whether the differences between the medians in both patches were statistically significant. The results obtained revealed statistically significant differences during the second year, at a 85% and 95% significance level, use being made of annual data and that recorded during the period of maximum interest, March-October, respectively. The decrease in soil respiration in the RT patch was around 24%. The second test was aimed at describing and comparing the influence of soil temperature on soil CO2 flux. By using the data of both patches recorded during the first year, an empirical equation on 10-cm soil temperature was fitted and tested on the data corresponding to the second year in each of the patches. Then, a comparison between the medians of the differences between the estimated and observed values was again performed by means of the Kruskal-Wallis test. The over-prediction of the model in the RT patch, statistically significant at a 90% significance level, was roughly 23%, confirming again the decrease in soil respiration one year after this agricultural management practice had been implemented. PMID- 12079079 TI - Suppression of nitrification and N2O emission by karanjin--a nitrification inhibitor prepared from karanja (Pongamia glabra Vent.). AB - A laboratory incubation study was undertaken to study nitirification and N2O emission in an alluvial, sandy loam soil (typic ustochrept), fertilized with urea and urea combined with different levels of two nitrification inhibitors viz. karanjin and dicyandiamide (DCD). Karanjin [a furanoflavonoid, obtained from karanja (Pongamia glabra Vent.) seeds] and DCD were incorporated at the rate of 5%, 10%, 15%, 20% and 25% of applied urea-N (100 mg kg(-1) soil), to the soil (100 g) adjusted to field capacity moisture content. Mean N2O flux was appreciably reduced on addition of the inhibitors with urea. Amounts of nitrified N (i.e. (NO3- + NO2-)-N) in total inorganic N (i.e. (NO3 + NO2- + NH4+)-N) in soil were found to be much lower on the addition of karanjin with urea (2-8%) as compared to urea plus DCD (14-66%) during incubation, indicating that karanjin was much more potent nitrification inhibitor than DCD. Nitrification inhibition was appreciable on the application of different levels of karanjin (62-75%) and DCD (9-42%). Cumulative N2O-N loss was found to be in the range of 0.5-80% of the nitrified N at different stages of incubation. Application of karanjin resulted in higher mitigation of total N2O-N emission (92-96%) when compared with DCD (60 71%). PMID- 12079080 TI - Dynamic-chemical coupling of the upper troposphere and lower stratosphere region. AB - The importance of the interaction between chemistry and dynamics in the upper troposphere and lower stratosphere for chemical species like ozone is investigated using two chemistry-climate models and a Lagrangian trajectory model. Air parcels from the upper troposphere, i.e. regions of lightning and aircraft emissions, are able to be transported into the lowermost stratosphere (LMS). Trajectory calculations suggest that the main transport pathway runs via the inter tropical convergence zone, across the tropical tropopause and then to higher latitudes, i.e. into the LMS. NOx from aircraft emissions at mid-latitudes are unlikely to perturb the LMS since they are washed-out while still in the troposphere. In contrast, NOx from tropical lightning has the chance to accumulate in the LMS. Because of the longer residence times of NOx in the LMS, compared to the upper troposphere, this excess NOx from lightning has the potential to form ozone in the LMS, which then is transported back to the troposphere at mid-latitudes. In the models, around 10% of the ozone concentration and 50% of the NOx concentration in the northern hemisphere LMS is produced by lightning NOx At least 5% of the ozone concentration and 35% the NOx concentration at 150 hPa at mid-latitudes originates from tropical lightning in the climate-chemistry simulations. PMID- 12079081 TI - Volatile organic compounds in some urban locations in United States. AB - Volatile organic compounds (VOCs) have been determined to be human risk factors in urban environments, as well as primary contributors to the formation of photochemical oxidants. Ambient air quality measurements of 54 VOCs including hydrocarbons, halogenated hydrocarbons and carbonyls were conducted in or near 13 urban locations in the United States during September 1996 to August 1997. Air samples were collected and analyzed in accordance with US Environmental Protection Agency-approved methods. The target compounds most commonly found were benzene, toluene, xylene and ethylbenzene. These aromatic compounds were highly correlated and proportionally related in a manner suggesting that the primary contributors were mobile sources in all the urban locations studied. Concentrations of total hydrocarbons ranged between 1.39 and 11.93 parts per billion, by volume (ppbv). Ambient air levels of halogenated hydrocarbons appeared to exhibit unique spatial variations, and no single factor seemed to explain trends for this group of compounds. The highest halogenated hydrocarbon concentrations ranged from 0.24 ppbv for methylene chloride to 1.22 ppbv for chloromethane. At participating urban locations for the year of data considered, levels of carbonyls were higher than the level of the other organic compound groups, suggesting that emissions from motor vehicles and photochemical reactions strongly influence ambient air concentrations of carbonyls. Of the most prevalent carbonyls, formaldehyde and acetaldehyde were the dominant compounds, ranging from 1.5-7.4 ppbv for formaldehyde, to 0.8-2.7 ppbv for acetaldehyde. PMID- 12079082 TI - Methane emissions from a coastal lagoon: Vembanad Lake, West Coast, India. AB - An attempt has been made to estimate methane fluxes from a tropical coastal wetland the Vembanad Lake, a lagoon along the West Coast of India. It has been found that Vembanad Lake contributes significant amount of methane to the atmosphere. Average emissions varied spatially within the lake. Methane emissions were 193.2 +/- 24.5 mg m(-2) h(-1) at Kumaragam (fresh water) as compared to 9.3 +/- 9.6 mgm(-2) h(-1) at Pullot (brackish water) site. Seasonal variation was significant between pre- and post-monsoons. Soil temperature, time of the day, salinity sediment organic carbon, all control the rate of methane emissions from the Vembanad Lake. PMID- 12079083 TI - Adolescent dating violence: prevalence and psychological outcomes. PMID- 12079084 TI - Date violence and date rape among adolescents: associations with disordered eating behaviors and psychological health. AB - OBJECTIVES: The goal of the study was to assess the prevalence of date violence and rape in adolescents, to examine associations between date violence and rape and disordered eating behaviors and psychopathology, and to determine if these associations remain significant after controlling for sociodemographic characteristics and other physical and sexual abuse by an adult. METHOD: A Minnesota school-based sample of 81,247 boys and girls in 9th and 12th grades completed the 1998 Minnesota Student Survey. RESULTS: Overall, approximately 9% of girls and 6% of boys had experienced date violence or rape. Significant differences across race and grade were found. Date violence and rape is associated with higher rates of disordered eating behaviors and suicidal thoughts and attempts, and lower scores on measures of emotional well-being and self esteem. Over 50% of youth reporting both date violence and rape also reported attempting suicide. Controlling for race and age, adolescents who have experienced both date violence and rape were more likely to use laxatives (OR: girls = 5.76; boys = 28.22), vomit (OR: girls = 4.74; boys = 21.46), use diet pills (OR: girls = 5.08; boys = 16.33), binge eat (OR: girls = 2.15; boys = 5.80), and have suicidal thoughts or attempts (OR: girls = 5.78; boys = 6.66) than their nonabused peers. These odds were weakened but remained significant after controlling for other abuse by an adult. Furthermore, a greater percentage of girls and boys who reported an abusive dating experience also reported repeat victimization (physical or sexual abuse perpetrated by an adult) when compared to their peers without an abusive dating experience. DISCUSSION: Abusive experiences during dating relationships may disrupt normal developmental processes, including the development of a stable self-concept and integrated body image during adolescence. This disruption manifests itself through thoughts, feelings, and behaviors. Further research should explore effects of adverse adolescent dating experiences. PMID- 12079085 TI - Pelvic fractures in infants as a sign of physical abuse. AB - OBJECTIVE: The purpose of this article is to describe pelvic fractures in two abused male infants, and to determine if the literature describes pelvic fractures in infants as suspicious for child physical abuse. RESULTS: Two infants are described with unexplained pelvic fractures. While in one case child abuse was obvious, careful attention to the radiological evaluation and case history led to the correct diagnosis of child abuse in the second case. A search for information regarding pelvic fractures in children resulted in information limited to the radiological and surgical literature. CONCLUSIONS: Medical providers frequently encounter children with fractures suspicious for child abuse. The most common fractures seen in abused children are metaphyseal, rib, skull, and long bone fractures. This report illustrates that pelvic fractures occurring in the absence of serious, well documented accidents should be considered highly suspicious for child physical abuse. PMID- 12079087 TI - Munchausen by Proxy in an international context. AB - OBJECTIVE: The professional literature on Munchausen by Proxy (MBP) abuse consists of more than 400 articles, chapters, and books. Most have come from a handful of English-speaking industrialized countries. Our aims were to establish the extent to which published work about MBP has emerged from outside these countries, and to determine the characteristics of any reported cases. METHOD: Numerous health care computer databases were queried, and the results supplemented by materials accumulated less formally. RESULTS: We identified 59 articles from 24 countries describing at least 122 cases in 9 different languages. Among cases in which the information was available, the mother was the sole perpetrator in 86%, the victim was aged between 3 years and 13 years in 52%, and the victim was male in 54%. The presentations of MBP appear to be similar across the world with the exception of induced apnea, which emerged as notably uncommon in this review. An extensive table presents the characteristics of each case. CONCLUSIONS: MBP clearly is not a phenomenon unique to Western or highly medicalized societies. This form of abuse is being increasingly recognized and reported throughout the world. The literature from the US, Canada, UK, Australia, and New Zealand often presupposes access to resources, such as subspecialists and social service agencies, that may be seriously constrained in other countries. The contributions of professionals elsewhere will be vital in ensuring that the efforts in English-speaking industrialized settings to develop standards of care encompass an international perspective. PMID- 12079086 TI - Different interpretations of Munchausen Syndrome by Proxy. AB - The definition of Munchausen Syndrome by Proxy is reviewed and considered in the context of the overlap with other harmful behaviors of parents. The high incidence of personal abnormal illness behavior in the perpetrators is leading to increasing concern about the safety of children who are cared for by parents who have abnormal illness behavior. PMID- 12079088 TI - Beyond collusion: active illness falsification. AB - OBJECTIVE: This article explores the relationship between factitious disorder by proxy victimization and the genesis of factitious disorder in young people. It is hoped that this will aid in our understanding of how some illness falsification behaviors may be learned and transmitted within the family system. METHOD: A discussion of the origins of adult factitious disorder and recent findings on the phenomenon of illness falsification in children and adolescents is integrated with some of the more ambiguous or "blended" cases which combine primary falsification by the youngster with caregiver collusion. Those less easily classified cases of factitious illness which fall in the gray areas, containing elements of both independent illness falsification by a child or adolescent with an earlier history of collusion with a parent's Munchausen by Proxy disorder, are proposed as a type of transitional case which may help us better understand the process by which illness falsification is learned. RESULTS: The literature on illness falsification in adults supports the possibility that adult factitious disorder may have its origins in adolescence or perhaps even earlier. Several cases are identified which suggest that some youngsters independently falsifying illness may have had earlier experiences of Munchausen by Proxy victimization or perhaps experienced the modeling or encouragement of illness falsification by a caregiver. Certain elements of the child victim experience, including efforts to overcome feelings of powerlessness, chronic lack of control, and disappointment in the physician are suggested as possible dynamics in the eventual development of independent illness falsification behaviors. CONCLUSIONS: The many unanswered questions in our understanding of the development of factitious illness in children and adolescents suggest avenues for further research. It is hoped that increased understanding will eventually allow more rapid, reliable identification of these patients and more effective interventions within the family system, with positive implications for future generations. PMID- 12079089 TI - On the importance of motivation in Munchausen by Proxy: the case of Kathy Bush. AB - The recent trial and conviction of Kathy Bush for abusing her daughter is used to illustrate (1) the nature of the motivation, in at least some cases of MBP, and (2) the importance of distinguishing the motivation found in MBP from that found in other forms of child abuse and other conditions involving factitious illness production. PMID- 12079090 TI - Vulnerability of groundwater to pollution from agricultural diffuse sources: a case study. AB - Kumluca, near Antalya in Turkey, is an important plain with its intensive agricultural activities employing greenhouses. The chemical fertilizer application practices caused excessive increases of the nitrogen, phosphorus and salinity within groundwater. A study has been initiated to assess the present state of the groundwater pollution problem of Kumluca Plain. A total of nine measurement and sampling stations have been selected to represent different depths groundwater table, different types of agricultural activities and different soil types. The magnitudes of the parameters: temperature, salinity and conductivity, nitrate, nitrite, ammonia, orthophosphate and fecal coliform were determined for groundwater. Soil samples collected from the stations have been analyzed for several parameters such as texture, total salinity, total nitrogen, and total phosphorus. The measurement and analyses results of the groundwater showed wide spatial variations depending on factors such as the quality of irrigation water, depth groundwater, soil characteristics, type and age of agriculture and hydrology. Groundwater vulnerabilities to pollution have been analyzed using the SEEPAGE Model approach. Furthermore the soil, aquifer and groundwater characteristics, which will be utilized to establish "cause" and "effect" relationships in future, have been clarified. PMID- 12079091 TI - Characterization and mass load estimates of organic compounds in agricultural irrigation runoff. AB - Investigations of agricultural chemicals in surface runoff typically target nutrients or specific pesticides; however, numerous other organic compounds are regularly applied to agricultural fields in pesticide formulations, irrigation water, soil amendments and fertilizers. Many of these compounds have toxicological significance. We conducted a broad spectrum analysis of surface runoff from individual irrigated agricultural fields in coastal southern California to characterize organic compounds amenable to analysis by gas chromatography-mass spectrometry and to estimate the mass flux of selected chemicals. Aqueous phase extracts contained several pesticides, as well as personal care product ingredients and pharmaceutically active compounds apparently derived from treated wastewater used for irrigation. Several compounds potentially associated with pesticide adjuvants were also present in aqueous phase extracts. Dissolved NOM constituents in water phase extracts included n fatty acids, aliphatic alcohols and plant terpenoids. Tentatively identified compounds sorbed to suspended particles included pesticides, a fecal sterol, aliphatic and alicyclic hydrocarbons, aliphatic alcohols, aldehydes, and C14 and C16 n-fatty acids and fatty acid esters. Bicyclic and polycyclic aromatic hydrocarbons were identified in both aqueous and suspended particle phases. Constituent concentrations, including total suspended solids (TSS), varied over the course of the sampled events by up to an order of magnitude, and typically were not correlated with flow. Variation in sorbed organic compound concentrations often did not parallel those for TSS concentration. Mass load estimates were strongly influenced by the choice of sampling interval. PMID- 12079092 TI - Fate of pesticides in soil in a coastal lagoon area and associated water quality impacts. AB - Pesticides used on agricultural lands are among the significant diffuse sources of pollutants. They are poisons and can be particularly dangerous when misused and/or applied without care. Although certain characteristics of pesticides are well known, their final characteristics after they reach a waterbody are extremely difficult to estimate. Regarding the soil and water environment, it is necessary to conduct both a field study and laboratory analysis. Such a study has been performed in the catchment area of Dalyan Lagoon, Turkey, joining the Mediterranean Sea, that was selected as the pilot region. The input of pesticides, transport in soil and transfer to the water environment together with their probable impacts on water quality are determined by conducting detailed field surveys and water residue analyses. Within the scope of the study, the behavior and fate of pesticides both in soil and water are investigated in detail. Water residue experiments are conducted on four sets of water samples representing the seasons of the year 1999-2000 for the pre-selected 6 pesticides. Two sets of water samples (surface and bottom) are taken from the 16 stations along the lagoon channel and two lakes within the catchment area. The results and findings show the significance of the use of pesticides as they exist in the water environment around and/or slightly above the limits stated in the national current regulations except dichlorvos which presents very high values compared to its tolerance limit. Of particular interest, the results are used to enlighten the farmers and the public and increase awareness of pesticides as contaminants in valuable coastal waters. PMID- 12079093 TI - Runoff characteristics of particulate pesticides in a river from paddy fields. AB - Runoff characteristics of particulate pesticides from paddy fields have been intensively observed in the Koise River in Japan. The 8 pesticides that are applied to paddy fields were analyzed in both particulate and dissolved forms. The concentrations and the detection frequencies of particulate pesticides were lower than those of dissolved pesticides. The particulate pesticide concentrations in the river water were evaluated based on the soil sorption coefficient, particulate organic carbon concentration, and dissolved pesticide concentrations. The particulate pesticide concentrations in the river were higher than evaluated concentrations because the paddy soil contained more pesticides than did suspended solids in the river water discharged during rain events, and because the desorption rates of pesticides were slow. In observations made during rains, the particulate pesticide concentrations increased with the increases in both the discharge rate and the concentrations of suspended solids. The particulate loading was slight compared with dissolved loading, but particulate pesticides may be influenced by enclosed areas such as a lake or estuary because under such conditions particulate matter settles vertically and the pesticide decomposition rate in sediment is slow compared with that in water. PMID- 12079094 TI - Runoff characteristics of pesticides from paddy fields and reduction of risk to the aquatic environment. AB - Simultaneous rice-transplanting and subsequent application of pesticides to the paddy fields in the catchment of a rural river on weekends from late May to mid June caused high concentrations and high loadings of pesticides in the river. Imperfections in irrigation water management and the overflow discharge from paddy fields during heavy storm events increased the risk of drinking water contamination and threatened the aquatic ecosystems. Several pesticides were detected at nearly ten ppb in the biofilms on the river bed. In order to reduce the risks due to runoff pesticides, it is necessary to improve the management of irrigation and to strengthen the biodegradation capacity of biofilms on the river bed by maintaining a long retention time in drainage channels. PMID- 12079095 TI - Elaboration of a complex GIS application in a catchment area. AB - Rearrangement of land resources after political changes has not yet been finished in Hungary. It is almost impossible to collect information necessary for planning activities on outer areas of settlements. The data are distributed among various organizations and can be found in diverse forms or there are no available data at all. However water quality protection has become legally ordered concerning municipal activities around Lake Balaton which is considered as the most important recreation area and tourist target in Hungary and is also affected by a number of factors providing sources of environmental conflicts. Settlements in a catchment area (Tetves Creek) on the southern shoreline of Lake Balaton in Central Hungary tendered a complex project for collecting sources of authentic data of the Hungarian rural areas along with systematizing and saving these data in a uniform GIS. An application using Autodesk MapGuide Program for Internet realization was developed. The implemented web-based system can be used in Internet and Intranet environments. PMID- 12079096 TI - Predicting pesticide concentrations in river water with a hydrologically calibrated basin-scale runoff model. AB - Hydrological diffuse pollution models require calibration before they can be used to make accurate long-term predictions for a range of hydrological and meteorological conditions. As such, the applicability of the models to the dispersion of new pesticides is limited due to the lack of calibration data. In this study, the performance of a GIS-based basin-scale runoff model for predicting the concentrations of paddy-farming pesticides in river water was examined when calibrated using hydrological data alone, without optimization based on empirical pesticide concentration data. The prediction accuracy on a daily or hourly scale was somewhat unsatisfactory due to inevitable compromises concerning rice farming schedules. However, the month-averaged pesticide concentrations were satisfactorily accurate; more than 50% of predicted values were between half and twice the observed values, considering the deficiencies of the input data, particularly for pesticide usage, which may include up to 50% error. PMID- 12079097 TI - Modeling diffuse pollution with a distributed approach. AB - The transferability of parameters for non-point source pollution models to other watersheds, especially those in remote areas without enough data for calibration, is a major problem in diffuse pollution modeling. A water quality component was developed for WATFLOOD (a flood forecast hydrological model) to deal with sediment and nutrient transport. The model uses a distributed group response unit approach for water quantity and quality modeling. Runoff, sediment yield and soluble nutrient concentrations are calculated separately for each land cover class, weighted by area and then routed downstream. The distributed approach for the water quality model for diffuse pollution in agricultural watersheds is described in this paper. Integrating the model with data extracted using GIS technology (Geographical Information Systems) for a local watershed, the model is calibrated for the hydrologic response and validated for the water quality component. With the connection to GIS and the group response unit approach used in this paper, model portability increases substantially, which will improve non point source modeling at the watershed scale level. PMID- 12079098 TI - Use of simulation mass balance modeling to estimate phosphorus and bacteria dynamics in watersheds. AB - Dynamic simulation technology is integrated with mass balance concepts and compartment-flux diagramming to create computer models that estimate contaminant export from watersheds over long and short-term futures under alternative simulated policies of watershed management. The Watershed Ecosystem Nutrient Dynamics (WEND) model, applied to developed watersheds with a mix of urban, agricultural, and forest land-uses, predicted phosphorus (P) export from three watersheds; a 275,000 ha dairy/urban watershed, a 77,000 ha poultry/urban watershed, and a 23,000 ha swine dominated watershed. Urban, agricultural, and forestry activities contribute to P export in different proportions. In all cases the P imports to the watershed exceed total export and P accumulates in watershed soils. Long-term future P export patterns are compared for several watershed management strategies that range from encouragement of rapid urban growth to aggressive environmental protection. The specific response of each watershed to imposed management is unique, but management strategies designed to reduce export of P over the long-term need to consider options that promote P input/output balance. Using this same approach, the Watershed Ecosystem Bacterial Dynamics (WEBD) model assesses the dynamics of bacterial populations in a small case-study watershed over an annual cycle as influenced by dairy farm management actions. WEND and WEBD models integrate the diversity of activities and stakeholders interested in the watershed and promote development of a more holistic understanding of watershed function. Model outputs are designed to assist watershed policy-makers, managers, and planners to explore potential future impacts of management/policy decisions. PMID- 12079099 TI - Stochastic modelling of phosphorus transfers from agricultural land to aquatic ecosystems. AB - This paper describes a simple model of phosphorus (P) transfer from agricultural land to surface waters which incorporates the effects of spatial variability in catchment properties and uncertainty in model parameter values. TOPMODEL concepts are used to estimate water, solute and sediment fluxes to water bodies. The model predicts the spatial distribution of water table depth and saturation-excess overland flow based on topography. Dissolved P (DP) transfer is assumed to occur vertically in the unsaturated zone and laterally in the saturated zone. Readily soluble P is assumed to decrease exponentially with soil depth. Particulate P (PP) transfers are modelled by estimating overland flow discharge and associated sediment transport capacity. Uncertainty in the distribution of soil surface P concentrations and model parameters controlling the mobility of soil P are incorporated using Monte Carlo simulation. Predicted losses of DP are well correlated with discharge and those of PP are episodic. Highest losses of P tended to be predicted near to the stream where the water table is close to the surface. The combination of a deterministic model core with a stochastic generation of model parameters or state variables provides an attractive way of embracing variability and uncertainty in models of this kind. PMID- 12079100 TI - SIMPLE: assessment of non-point phosphorus pollution from agricultural land to surface waters by means of a new methodology. AB - In the past, environmental Phosphorus (P) parameters like soil P indices have been used to catogorize the potential risk of P losses from agricultural land. In order to assess the actual risk of P pollution of groundwater and surface waters, dynamic process oriented soil and water quality models have been frequently used. Recently, an approximating model for phosphorus, called SIMPLE, has been developed. This model approximates the output from a complex dynamic water quality model. The approximating model is called a metamodel. This simple P-model proves to be a powerful tool for quick assessment of the risk of P pollution from agricultural land to surface waters. PMID- 12079101 TI - Quantitative comparison of forests and other areas with dry weather input loading in the Lake Biwa catchment area. AB - For the evaluation of pollutants loading to Lake Biwa, comprehensive river surveys on river mouths and forest sites were conducted 9 times from 1996 to 1999, on 25-40 main rivers in the Lake Biwa watershed. The main results obtained are as follows. (1) River water quality reflects regional characteristics a of their catchment areas, and the concentration difference among rivers ranged between 2-3 fold. (2) Concentration variation shows different patterns with time and location depending on water quality indices a used. (3) Indices related to organic matter and nutrients have lower correlation between forests and river mouths, but those related to ionic species showed strong correlation. (4) Flux comparison of forest and river mouth sites suggests that pollutants from catchment areas are conveyed to the lake not only through rivers but also underground. (5) In dry weather conditions, forests contribute 30% to the whole pollutants (TN, TP, and TCOD(Mn)) loading, and the remainder is derived mainly from paddy fields and residential/commercial zones. (6) Unit loading factors from forests are estimated as 640, 57 and 1200 kg/km2/y, respectively for TN, TP and TCOD(Mn), while those from other areas are estimated as 2,500, 208 and 4,200 kg/km2/y. PMID- 12079102 TI - Nitrate in aquifers beneath agricultural systems. AB - Research from several regions of the world provides spatially anecdotal evidence to hypothesize which hydrologic and agricultural factors contribute to groundwater vulnerability to nitrate contamination. Analysis of nationally consistent measurements from the U.S. Geological Survey's NAWOA program confirms these hypotheses for a substantial range of agricultural systems. Shallow unconfined aquifers are most susceptible to nitrate contamination associated with agricultural systems. Alluvial and other unconsolidated aquifers are the most vulnerable and shallow carbonate aquifers provide a substantial but smaller contamination risk. Where any of these aquifers are overlain by permeable soils the risk of contamination is larger. Irrigated systems can compound this vulnerability by increasing leaching facilitated by additional recharge and additional concentrations of groundwater nitrate than all other agricultural systems, although mean nitrate concentrations in counties with dairy, poultry, cattle and grains, and horticulture systems were similar. If trends in the relation between increased fertilizer use and groundwater nitrate in the United States are repeated in other regions of the world, Asia may experience increasing problems because of recent increases in fertilizer use. Groundwater monitoring in Western and Eastern Europe as well as Russia over the next decade may provide data to determine if the trend in increased nitrate contamination can be reversed. If the concentrated livestock trend in the United States is global, it may be accompanied by increasing nitrogen contamination in groundwater. Concentrated livestock provide both point sources in the confinement area and intense non-point sources as fields close to facilities are used for manure disposal. Regions where irrigated cropland is expanding, such as in Asia, may experience the greatest impact of this practice. PMID- 12079103 TI - Evaluation of efficiencies of diffuse allochthonous and autochthonous nutrient input control in restoration of a highly eutrophic lake. AB - Mogan Lake is an important recreational area for Metropolitan Ankara-Turkey. It is a shallow eutrophic lake with a dense growth of macrophytes. The main contributors of nutrients and other pollutants to the lake are the creeks carrying the runoff water from the watershed and upland farming land, in addition to the domestic and industrial wastewater discharges from a nearby town and industries. Hydrodynamic and water quality modeling techniques were used to determine the optimum management schemes for the lake restoration and diffuse pollution control. Management scenarios were devised and tested to control allochthonous and autochthonous nutrient inputs to the lake. Phosphorus and nitrogen load reductions were the main test elements for the control of allochthonous nutrient inputs. The scenario analysis revealed that reduction of phosphorus and nitrogen loads from diffused sources will have a marginal effect on controlling eutrophication if macrophyte growth is left uncontrolled. Scenarios employing macrophyte harvesting and sediment dredging have been evaluated for autochthonous nutrient input control. Sediment dredging alone has been shown to yield the most favorable conditions for water quality improvement in Mogan Lake. Further, control of diffuse pollution was an essential final step to achieve an acceptable long-term sustainable water quality improvement in the lake. PMID- 12079104 TI - Real-time water quality monitoring and regression analysis to estimate nutrient and bacteria concentrations in Kansas streams. AB - An innovative approach currently is underway in Kansas to estimate and monitor constituent concentrations in streams. Continuous in-stream water-quality monitors are installed at selected U.S. Geological Survey stream-gaging stations to provide real-time measurement of specific conductance, pH, water temperature, dissolved oxygen, turbidity, and total chlorophyll. In addition, periodic water samples are collected manually and analyzed for nutrients, bacteria, and other constituents of concern. Regression equations then are developed from measurements made by the water-quality monitors and analytical results of manually collected samples. These regression equations are used to estimate nutrient, bacteria, and other constituent concentrations. Concentrations then are available to calculate loads and yields to further assess water quality in watersheds. The continuous and real-time nature of the data may be important when considering recreational use of a water body; developing and monitoring total maximum daily loads; adjusting water-treatment strategies; and determining high constituent concentrations in time to prevent adverse effects on fish or other aquatic life. PMID- 12079105 TI - Application of an automatic sampling and measurement system to a mountainous stream investigation during rain events. AB - An automatic sampling and measurement system was developed to take water samples efficiently during rain events. The system consisted of an automatic sampler, a rain gauge and a data logger as well as sensors for pH, electrical conductivity (EC), and both water and air temperature. The system was tested in a stream in a forested watershed (5.8 km2) located in the middle of Hyogo prefecture, Japan. The sampling program has been improved gradually. For several rain events of 30 to 157 mm, most water samples were in agreement with the hydrograph from the beginning of each rainfall until the rain had stopped and the water level had begun to fall. The fluctuations in water quality in the samples taken by the automatic sampler during those rain events showed patterns similar to those of water samples taken by hand. There were also no problems with the water level or the EC sensor during the investigation periods, but the pH values were lower than those in the laboratory. The results showed that the system is suitable for taking water samples from mountainous streams during rain events. PMID- 12079107 TI - Search for the northwest passage: the assignation of NSP (non-point source pollution) rights in nutrient trading programs. AB - The search for solutions to the problem of non-point source pollution (NSP) includes alternatives based on theories associated with the use of tradable pollution permits. Tradable permit programs have received significant support as a promising policy for the reduction of effluent discharges but programs in practice have not been regarded as successful. The lack of success is ascribed to the design of the programs. However, this may be a design problem which is insurmountable due to the nature of the NSP problem. Tradable permit solutions are based on an assumption that the assignation of quantifiable rights to both point and nonpoint sources, based on some predetermined ambient water quality measure, is possible. The conclusion here is that there are significant features particular to NSP that hinder the introduction of rights and significantly decrease the utility of tradable permit solutions. PMID- 12079108 TI - Willingness to pay for flood and ecological risk reduction in an urban watershed. AB - Urban watershed managers frequently must address alternative policy goals; flood control and ecological risk reduction. This study combines hydrologic models of flood control and biotic models of ecologic risk with economic models of willingness-to-pay and psychological models of risk processing and planned behavior to evaluate these two alternative policy objectives. The findings reveal that flood risk exposure, especially for those individuals who would remain outside the 100 year flood plain if the project were enacted, does influence the financial support that local residents would be willing to make to a flood control project. Other important determinants include demographic factors such as income, and attitudinal measures of the respondent. Expanding the scope of the project to include ecological risk reduction does not, however, appear to change the average willingness-to-pay for a project. PMID- 12079106 TI - Impact of non-detects in water quality data on estimation of constituent mass loading. AB - Often, fractions of stormwater constituents are not detected above laboratory reporting limits and are reported as non-detect (ND), or censored data. Analysts and stormwater modelers represent these NDs in stormwater data sets using a variety of methods. Application of these different methods results in different estimates of constituent mean concentrations that will, in turn, affect mass loading computations. In this paper, different methods of data analysis were introduced to determine constituent mean concentrations from water quality datasets that include ND values. Depending on the number of NDs and the method of data analysis, differences ranging from 1 to 70 percent have been observed in mean values. Differences in mean values were, as shown by simulation, found to have significant impacts on estimations of constituent mass loading. PMID- 12079109 TI - Nitrate leaching from construction sites to groundwater in the Nottingham, UK, urban area. AB - Nitrate pollution has been identified as a major water quality issue in the UK. The aim of this project is to research the rate of nitrate leaching to groundwater that arises from construction works. The study area is situated in Nottingham UK, which is situated on the Triassic Sandstone aquifer. Soil samples up to a depth of 2.50 m were taken from three sites under construction and other land use. The results have shown a high variability in the concentrations of soil nitrate. The reasons for this variability include soil type, past land use, soil treatment and type of vegetation prior to construction works. The average nitrogen load was 65 kg N ha(-1) which is higher than the nitrate leaching observed when temporary grassland is ploughed during autumn. The highest nitrate concentrations were observed in an allotment site (133 kg N ha(-1)) due to the high amount of manure applied at this location. The construction practice of top soil stripping can produce a reduction of nitrate leaching because it removes the part of the soil that contains most of the potentially mineralizable nitrogen. PMID- 12079110 TI - Characterization of surface runoff in urban areas. AB - Water quality measurements of surface runoff have been carried out in selected residential and industrial zones in urban areas, in which yearly mean precipitation is 1,225 mm. The concentrations of constituents in the surface runoff were measured at sampling sites categorized by land use type in the residential zone, and by industry type in the industrial zone. The water quality constituents of BOD5, COD, SS, NO3-N, TKN, PO4-P, TP, n-Hexane extracts, Cr, Cu, Pb and Fe were analyzed. The event mean concentrations (EMCs) of COD, SS, TKN and TP in the residential zone were 313 mg/L, 279 mg/L, 8.45 mg/L, 1.98 mg/L, and those in the industrial zone were 80 mg/L, 106 mg/L, 5.07 mg/L, and 1.93 mg/L, respectively. Cumulative load curves were created to analyze the first-flushing effect of each pollutant related to the pollutant, the rainfall event, and the land use type. No general relationship between the cumulative load and runoff has been established. The degree of first-flushing effect by constituents was in the following order; TKN>COD>SS>HEM>TP>PO4-P. The correlations between SS and other constituents were analyzed to evaluate the efficiency of the physical treatment process to control the surface runoff in urban areas. Based on the correlation of constituents with SS, high treatment efficiency of SS, heavy metals, organic matter, and TP was expected. The unit pollutant loading rates of COD, SS, TKN, TP, Cr and Pb in the residential zone were 2,392, 2,130, 64.6, 15.1, 0.31, and 1.83 kg/ha/yr, and those in the industrial zone were 612, 812, 38.7, 14.8, 0.51 and 0.82 kg/ha/yr, respectively. PMID- 12079111 TI - Comparison of the pollutant loads in dry and wet weather runoff in a southern California urban watershed. AB - This research compares the relative contributions of potential contaminants discharged in dry weather flow (DWF) and wet weather flow (WWF) from the highly urbanized Ballona Creek watershed (BCW) in southern California using empirical and deterministic models. These models were used to compare the loading of the following pollutants: total suspended solids, biochemical oxygen demand, total nitrogen, total inorganic nitrogen, total Kjeldahl nitrogen, total phosphorus, copper, lead, arsenic, nickel, cadmium, and chromium. The results indicate DWF contributes approximately 10-30% of the total annual flow discharged from Ballona Creek. The annual DWF volume was fairly consistent; the variation in DWF percentage contribution was dependent on the highly variable volume of WWF. The relative contribution to the annual pollutant load varied considerably between each pollutant. In general, the DWF load was found to be significant, especially in years with lower precipitation totals. The results from this investigation have identified the relative relationship between DWF and WWF loads in the BCW and will aid in the decision-making process during the development of an integrated DWF-WWF management plan and allocation of water pollution control funds between DWF and WWF management. PMID- 12079112 TI - Evaluation of urban non-point source runoff of hazardous metals entering Santa Monica Bay, California. AB - Significant amounts of non-point source runoff enter the Santa Monica Bay from the Ballona Creek Watershed during wet weather flow. The primary objective of this study was to determine the concentrations of hazardous metal pollutants associated with the aqueous and suspended solid phases during wet weather flow. A calculation of the mass of the pollutants for each storm was conducted. Other objectives of this study were to evaluate during a storm event the relationships between (1) soluble and sorbed metals, (2) storm flow and pollutant loading, including a determination if a first flush was present, and (3) total mass loading of pollutants and relative pollution loading from three watershed sub basins. This study focused on the urbanized watershed of Ballona Creek, which is approximately 330 km2 (127 miles2) in size and developed primarily with residential and light commercial industries. In the 1997-98 rain season, an El Nino year, two storm events were monitored by the collection of hourly grab samples. An initial storm monitored Ballona creek, while a second storm also monitored Ballona Creek along with two smaller sub-watersheds, Centinela Channel and Sepulveda Channel. The results indicated the suspended solids phase primarily transported the mass for five of the six hazardous metals studied: cadmium, chromium, copper, lead, and nickel. Arsenic was found primarily in the aqueous phase. PMID- 12079113 TI - Evaluation of land use/land cover datasets for urban watershed modeling. AB - Land use/land cover (LULC) data are a vital component for nonpoint source pollution modeling. Most watershed hydrology and pollutant loading models use, in some capacity, LULC information to generate runoff and pollutant loading estimates. Simple equation methods predict runoff and pollutant loads using runoff coefficients or pollutant export coefficients that are often correlated to LULC type. Complex models use input variables and parameters to represent watershed characteristics and pollutant buildup and washoff rates as a function of LULC type. Whether using simple or complex models an accurate LULC dataset with an appropriate spatial resolution and level of detail is paramount for reliable predictions. The study presented in this paper compared and evaluated several LULC dataset sources for application in urban environmental modeling. The commonly used USGS LULC datasets have coarser spatial resolution and lower levels of classification than other LULC datasets. In addition, the USGS datasets do not accurately represent the land use in areas that have undergone significant land use change during the past two decades. We performed a watershed modeling analysis of three urban catchments in Los Angeles, California, USA to investigate the relative difference in average annual runoff volumes and total suspended solids (TSS) loads when using the USGS LULC dataset versus using a more detailed and current LULC dataset. When the two LULC datasets were aggregated to the same land use categories, the relative differences in predicted average annual runoff volumes and TSS loads from the three catchments were 8 to 14% and 13 to 40%, respectively. The relative differences did not have a predictable relationship with catchment size. PMID- 12079114 TI - Critical issues for stormwater ponds: learning from a decade of research. AB - The Queen's University/National Water Research Institute Stormwater Quality Enhancement Group has been actively researching stormwater ponds for the past decade, using a fully instrumented on-line system in Kingston, Ontario, Canada as a representative field installation of this group of stormwater best management practices, along with comprehensive surveys of other facilities as well. From this body of research, the Group has concluded that there are a number of identifiable factors, termed critical issues, which will significantly influence the success, failure and sustainability of these BMPs. Such factors will be important to a very diverse group of stakeholders in stormwater management, including designers, owners/operators, regulatory authorities and the general public. These factors can be grouped within the categories of initial design, operation and maintenance, performance and adaptive design. From this work, it is concluded that the so-called first generation quantity-control ponds may be outdated today, compared with the modern focus on quantity and quality issues in the second generation systems; nonetheless, without consideration of these critical issues and flexible design practices which can account for emerging or future issues, the current systems also run the risk of becoming outdated before the end of their design lives. PMID- 12079115 TI - The SOIL-N/WEKU model system--a GIS-supported tool for the assessment and management of diffuse nitrogen leaching at the scale of river basins. AB - The SOIL-N/WEKU model system was developed to estimate groundwater-borne nitrogen inputs into river systems. The core of this model system is composed of a soil nitrogen leaching model (SOIL-N) and a groundwater residence time/denitrification model (WEKU). The application of the model system was carried out in the framework of the EU-project RANR (Regional analysis of subsurface nitrogen retention and its impact on the nitrogen export from land to sea) for a macroscale study river basin in Germany (the Uecker basin, ca. 2,400 km2) and a mesoscale study catchment area in Denmark (the Gjern basin, ca. 200 km2). For both catchment areas, the modelled average nitrogen loads leached into the groundwater were about 40 kg N/ha a, while the remaining groundwater-borne nitrogen intake to rivers was quantified to an average of about 2 kg/ha a. The comparison with observed groundwater-borne riverine nitrogen loads showed a very good agreement, proving the key role nitrogen retention in groundwater plays in the two catchment areas. PMID- 12079116 TI - Nitrate pollution in groundwater and strategies to reduce pollution. AB - The input-intensive rainfed tropical ecosystem, where wet season (WS) rice (Oriza sativa L.)-dry season (DS) diversified high-value upland crops like vegetables predominate, has resulted in a problem of a large leakage of N into the environment, thereby polluting the water. Excessive use of N fertilizer in high value crops grown in DS is economically motivated. Out of twenty water sources evaluated in a watershed with a total area of 265 ha located in Magnuang, Ilocos Norte, Philippines, twelve had near or above the World Health Organization's (WHO) NO3-N limit for drinking water of 10 ppm. Soil mineral N (upper 100 cm) observed in seven rice-sweet pepper (Capsicum annuum L.) farmers' fields ranged from 111 to 694 kg ha(-1) which decreased by 10 to 68% in plots with dry-to-wet (DTW) crops like indigo, indigo+mungo and corn. In fallow plots where mineral N was either maintained or increased, it showed movement to lower soil profiles demonstrating NO3 leaching without a crop. On average, maize (Zea mays L.) captured 176 kg N ha(-1) and indigo (Indigofera tinctoria L.) 194 kg N ha(-1). In both fallow and planted plots, mineral N declined to low levels at 100% water filled pore spaces (WFPS) before rice transplanting. A strategy for including indigo plus maize as a N-catch crop is proposed to decrease NO3 leaching and maximize N use efficiency in a rice-sweet pepper cropping system. PMID- 12079117 TI - Integrated watershed approach in controlling point and non-point source pollution within Zelivka drinking water reservoir. AB - An agricultural watershed involves manipulation of soil, water and other natural resources and it has profound impacts on ecosystems. To manage these complex issues, we must understand causes and consequences and interactions-related transport of pollutants, quality of the environment, mitigation measures and policy measures. A ten year period of economic changes has been analysed with respect to sustainable development concerning Zelivka drinking water reservoir and its watershed, where agriculture and forestry are the main human activities. It is recommended that all land users within a catchment area should receive payments for their contribution to water cycle management. Setting up the prevention principles and best management practices financially subsidized by a local water company has been found very effective in both point and non-point source pollution abatement, and the newly prepared Clean Water Programme actively involves local municipal authorities as well. The first step based on systems analysis was to propose effective strategies and select alternative measures and ways for their financing. Long term monitoring of nutrient loads entering the reservoir and hazardous events statistics resulted in maps characterising the territory including vulnerable zones and risk factors. Financing involves providing annual payments to farmers, who undertake to manage specified areas of their land in a particular way and one-off payments to realise proposed issues ensuring soil conservation and watershed ecosystem benefits. PMID- 12079118 TI - Development of a new mathematical model for subsurface water quality management. AB - The interfaces between free (e.g., groundwater) and porous (e.g., soil) flow zones in the subsurface represent important transition zones across which many important transfer/exchange processes occur. The understanding of these interactive phenomena and the way these regions behave in combination is, therefore, critical for management of subsurface water quality. Indispensable to this is numerical modelling and simulation as they can handle complex flow domains and minimise the analysis cost and time. In the present work, the hydrodynamic conditions for a combined free and porous flow domain in the subsurface are analysed. An investigation into the fluid dynamical behaviour for different aspect ratios of the domains is of most interest. PMID- 12079119 TI - Nutrient management for coastal zones: a case study of the nitrogen load to the Stockholm Archipelago. AB - This study investigates cost-effective solutions of decreasing the nutrient load to a coastal area, using a drainage basin approach. The study is applied to the Stockholm Archipelago, a coastal area of the Baltic Sea suffering from eutrophication caused by the load of nutrients entering the area. Nitrogen is the nutrient of concern in this study since it is the limiting nutrient of the Archipelago. The main sources of nitrogen are wastewater plants, agriculture, and atmospheric depositions. The final impact of a deposition depends on the buffering capabilities it is subject to on its trajectory from the source to the recipient. This is the reason for using a recipient oriented approach, in which the focus is to reduce the final impact of a deposition. The model integrates data over hydrology, land cover, land use, and economy in order to find the optimal allocation of measures. Results indicate that in order to achieve cost effectiveness, the major part of nitrogen load reduction to the Archipelago should be done at the wastewater plants and by constructing wetlands. The minimum annual cost of reaching a 50% reduction of the load to the Archipelago was estimated to around 191 million Swedish crowns (around $19 million). PMID- 12079120 TI - Development of a risk-based TMDL assessment approach using the integrated modeling system GIBSI. AB - Using the integrated modeling system GIB SI and a case study, this paper presents the development of a risk-based TMDL assessment approach that links wet (nonpoint/diffuse) and dry weather (point) sources to a probability of exceeding water quality standards (WQS) governing wateruses. The case study focused on determining whether WQS defining recreational uses of water requiring direct and prolonged contact were attainable if the waste water effluent of a small town was treated using aerated lagoons and if the agricultural nonpoint source (NPS) loads were reduced using different fertilization rates. Dry weather sources were assumed to solely contribute to bacteriological impairment of the studied river reach. Meanwhile, both wet and dry weather sources were assumed to contribute to aesthetic impairment. Simulation results showed that treating the waste water effluent while reducing the agricultural NPS loads by 27% allowed on average over a four-year study period for attainment of the bacteriological WQS 100% of the summer time while lowering the probability of exceeding the aesthetic WQS from 0.32 to 0.19 (30 to 18 days). The results of this study showed this risk-based assessment approach was well suited to establish TMDL. These probabilities should be evaluated using long meteorological series. PMID- 12079121 TI - Sensitivity analysis and auto-calibration of an integral dynamic model for river water quality. AB - ESWAT--Extended Soil and Water Assessment Tool--was developed to allow for an integral modelling of the water quantity and quality processes in river basins. ESWAT is a physically based, semi-distributed model, with a moderate-to-large number of parameters and input and output variables (depending on the desegregation scheme). An auto-calibration procedure was implemented for the optimisation of the process parameters. The procedure is based on a new approach for multi-objective calibration and incorporates the algorithms of the Shuffled Complex Evolution Method. The optimisation uses a global optimisation criterion, whereby several output variables can be taken into account simultaneously. A statistical method enables the aggregation of the objective functions for individual variables, hereby avoiding the weighting problem. To select the important parameters for the optimisation, a sensitivity analysis precedes the calibration. The latter analysis is based on the One-factor-At-a-Time (OAT) design approach. The sensitivity analysis and the calibration procedure are applied to the river Dender in Belgium. The river is characterised by high pollution loads and long residence times in summer periods. PMID- 12079122 TI - Geochemical and isotopic tracing of water in nested southern Minnesota corn-belt watersheds. AB - Land-use changes over the last century in southern Minnesota have influenced riverine water chemistry. A nested watershed approach was used to examine hydrologic pathways of water movement in this now agriculturally intensive region. From field scale subsurface tile-drains of the Beauford ditch to the respective outlets of the Cobb River and Blue Earth River, more than 125 samples were collected for major dissolved ions and isotopes between March 1994 and June 1996 over a range of climatic conditions that included snowmelt and storm-flows. Results indicate that riverine water chemistry is dominated by subsurface tile drained row crop agriculture. In the mid-1990s, regional ground water discharge into the Cobb and Blue Earth Rivers comprised less than 10% of the total flow based on ionic mixing calculations. Ammonia, present in manure or as anhydrous, is readily exchanged in the soil. This ion exchange releases increasing ratios of magnesium, sodium and strontium relative to calcium, the dominant cation. Soil thaw and snowmelt recharge influenced March-April tile-drain and ditch water isotopic values. Light deltaD values increased as spring infiltration-derived water was displaced from the soil zone by heavier summer precipitation. Delta15N followed a similar but opposite pattern with relatively heavy March-April tile drain and ditch values trending to lighter delta15N through the growing season. The future of southern Minnesota riverine water quality is closely linked to the management of the landscape. To improve the riverine environment, land owners and managers will need to address cropping systems, fertilization practices and drainage. PMID- 12079123 TI - Implementation of best management practices in agriculture: modelling and monitoring of impacts on nitrogen leaching. AB - In Kattegat and the coastal water of the Baltic Sea, high nitrogen input from agricultural land is considered to be the main reason for eutrophication. International agreements and governmental programs have set a target to reduce the anthropogenic nitrogen load by 50 percent. Improved nitrogen removal in treatment plants and efforts in agriculture have so far not decreased nitrogen transport to a sufficient extent. In this project the impact of agricultural practices on nitrogen leaching was investigated in two small agricultural catchments in Southwest Sweden. The root-zone leaching was estimated by an indexing technique. Simultaneously the transports in the stream outlets were monitored. During 1995 and 1999 the agricultural practices in the catchments were surveyed. Field data from the first survey indicated that fertilisation did not always match crop requirements, the area of undersown catch crop can be increased and autumn cultivation can be reduced. The second survey was preceded by an advisory campaign where each farmer was visited and presented with an environmental plan including fertilisation, cultivation, and crop rotation for the farm. The plan summarised the best management practices that could be realised under actual conditions. Results from the second survey showed that some changes in the agricultural practices were carried out after the advisory campaign. The nitrogen leaching from the root-zone was then estimated by the indexing technique, both for the time before and after the advisory program. The results showed that the estimated nitrogen leaching, as a mean value for 330 fields, decreased from 53 to 50 kgN ha(-1), due to adjustments of the agricultural practices. Monitoring of stream transports showed values of the same magnitude after correction for retention and other sources. In this short time perspective, decreases in transport due to changes in agricultural practices could not be separated from influence of weather conditions. In comparison to results from the Swedish monitoring programme for agriculture, the measured transports were normal for the region, where annual variation in precipitation and runoff is large. Theoretically, nitrogen leaching could be reduced by one third without any major economic constraints for the farmers. In general, the farmers were positive to advice and willing to try new farming techniques even if some measurements were not fully implemented during the investigation period. PMID- 12079124 TI - Phosphorus reductions following riparian restoration in two agricultural watersheds in Vermont, USA. AB - Achievement of management goals for Lake Champlain (Vermont/New York, USA and Quebec, Canada) will require significant reductions of phosphorus (P) loads from agriculture, the dominant diffuse source in the basin. Cost-effective P reduction strategies must be based on reliable treatment techniques beyond basic erosion control and animal waste storage practices. The Lake Champlain Basin Agricultural Watersheds National Monitoring Program (NMP) Project evaluates the effectiveness of low-cost livestock exclusion, streambank protection, and riparian restoration practices in reducing concentrations and loads of diffuse-source pollutants from grazing land at the watershed level. Treatment and control watersheds in northwestern Vermont have been monitored since 1994 according to a paired watershed design. Monitoring includes continuous stream discharge recording, flow proportional sampling for total P and other pollutants, and documentation of land use and agricultural management activities. Strong statistical calibration between the control and treatment watersheds has been achieved. Landowner participation in the land treatment program was entirely voluntary and all treatments were 100% cost-shared by the project and cooperators. Installation of riparian fencing, alternative water supplies, protected stream crossings, and streambank bioengineering was completed in 1997 at a cost of less than US$40,000. The paired-watershed design was effective in controlling for the influence of extreme variations in precipitation and streamflow over six years of monitoring. Two years of post-treatment data have documented significant reductions in P concentrations and loads from both treated watersheds. Reductions of approximately 20% in mean total P concentration and approximately 20-50% in mean total P load have been observed, with greater reductions occurring in the watershed receiving more extensive treatment. The effectiveness of riparian zone restoration in P reduction tended to be lower during periods of very high runoff, especially outside the growing season. PMID- 12079125 TI - Hydrogeological constraints on riparian buffers for reduction of diffuse pollution: examples from the Bear Creek watershed in Iowa, USA. AB - Riparian Management Systems (RiMS) have been proposed to minimize the impacts of agricultural production and improve water quality in Iowa in the Midwestern USA. As part of RiMS, multispecies riparian buffers have been shown to decrease nutrient, pesticide, and sediment concentrations in runoff from adjacent crop fields. However, their effect on nutrients and pesticides moving in groundwater beneath buffers has been discussed only in limited and idealized hydrogeologic settings. Studies in the Bear Creek watershed of central Iowa show the variability inherent in hydrogeologic systems at the watershed scale, some of which may be favorable or unfavorable to future implementation of buffers. Buffers may be optimized by choosing hydrogeologic systems where a shallow groundwater flow system channels water directly through the riparian buffer at velocities that allow for processes such as denitrification to occur. PMID- 12079126 TI - Effect of a cold-climate buffer zone on minimising diffuse pollution from agriculture. AB - Vegetative buffer zones adjacent to streams can filter diffuse pollution from agriculture. Under Nordic climatic conditions, major runoff from agriculture occurs during winter and especially during snowmelt. Field experiments documenting runoff and retention processes in buffer zones during winter conditions are essential. Agricultural runoff and buffer zone retention of nutrients and particles during winter and summer are compared. The study is based upon 8 years of data collected from 5 to 10 m wide buffer zones compared to plots without buffer zones. Volume proportional samples were collected after each runoff episode. Results show that the difference between precipitation and runoff is much higher during summer than winter, due to higher evapotranspiration and infiltration during the summer. Over 90% of total particle and phosphorus runoff occurred during winter. There was no significant difference in retention of particles and particle-bound nutrients during winter compared to summer. Sedimentation is one of the most important retention processes in surface runoff buffer zones. High retention during winter is probably caused by higher surface runoff and erosion. High surface runoff and flow velocity could cause erosion of coarser particles, which were more easily trapped in the buffer zone. High particle concentration in the runoff water may also enhance the aggregation of fine particles. PMID- 12079127 TI - Design considerations for increased sedimentation in small wetlands treating agricultural runoff. AB - Some suggestions to increase the sedimentation of non-point source pollution in small surface flow wetlands are presented. The recommendations are based on results from seven Norwegian constructed wetlands (CWs) after 3-7 years of investigation, and a literature review. The wetlands were located in first and second order streams. Surface areas were 265-900 m2, corresponding to 0.03-0.4% of the watershed. Each CW had a volume proportional composite sampler in the inlet and outlet, in addition to sedimentation plates. The mean annual retention of soil particles, organic particles and phosphorus was 45-75%, 43-67% and 20 44%, respectively. Results showed that erosion and transportation processes in arable watersheds influenced the retention. Sedimentation was the most important retention process, and increased with runoff, because the input of larger aggregates increased. Retention of nitrogen did not follow the same pattern, and was only 3-15%. Making CWs shallow (0-0.5 m) can optimize sedimentation. The hydraulic efficiency can be increased by aquatic vegetation, large stones in the inlet, baffles and water-permeable, low dams. Vegetation makes it possible to utilize the positive effect of a short particle settling distance, by hindering resuspension of sediments under storm runoff conditions. As a result, the phosphorus retention in shallow CWs was twice that of deeper ponds. PMID- 12079129 TI - SOIL and SOIL-NO at catchment scale--a case study for an agriculture-dominated catchment. AB - A one-dimensional model, SOIL/SOIL-NO, is used to simulate the water and nitrogen balance at catchment scale. The objective is to use the model in simulating the effects of best management practices on nitrogen runoff. The model was applied to individual farm fields in an agriculture dominated catchment and simulations were carried out for the period from 94-98. The results were promising. The simulated nitrogen runoff agreed quite well with the measured nitrogen loss at the main station. The effects of best management practices like optimal fertiliser application, catch crops and irrigation were simulated. For this particular catchment, the introduction of catch crops resulted in the best effect on reducing nitrogen runoff from agricultural dominated catchments. PMID- 12079128 TI - Wetland management to reduce Baltic Sea eutrophication. AB - Seven regions with coastal eutrophication problems in the Baltic Sea, including the Kattegat, constitute the BERNET project (Baltic Eutrophication Regional Network). To counteract eutrophication and associated severe biological conditions the countries around this large brackish water body must all cooperate. The regions are characterized by large differences in land use, e.g. agricultural intensity, and losses of retention capacity in the catchments due to wetland reclamation. Initially it has been necessary to identify nutrient sources -especially nitrogen--and technical, economical and even administrative obstacles to initiate eutrophication management measures. Nitrogen retention in different types of wetlands in the Baltic Sea Region has been analysed. The wetlands generally have a positive effect on reduced nitrogen transport to aquatic environments and it is generally accepted that measures leading to decreased losses of nutrients to the aquatic environment must be combined with measures leading to increased retention of nutrients in catchments. Data analysed in the BERNET project show that the potential for such a measure is large. Therefore, conservation and restoration initiatives for wetlands is an essential part of the work in the BERNET project. Wetlands have been drained or totally eliminated due to intensive agriculture in some regions while large scale rehabilitation of wetlands occurs in regions with less intensive agriculture. Based on land use data from the seven regions, the working group for wetland management within the BERNET project has identified the possible use of wetlands as building blocks as a contribution to the management of the Baltic Sea eutrophication. Several recommendations are presented on the wise use of existing and constructed wetlands for water quality management in relation to non-point nutrient pollution. PMID- 12079130 TI - Potential toxicity of pesticides measured in midwestern streams to aquatic organisms. AB - Society is becoming increasingly aware of the value of healthy aquatic ecosystems as well as the effects that man's activities have on those ecosystems. In recent years, many urban and industrial sources of contamination have been reduced or eliminated. The agricultural community also has worked towards reducing off-site movement of agricultural chemicals, but their use in farming is still growing. A small fraction, estimated at <1 to 2% of the pesticides applied to crops are lost from fields and enter nearby streams during rainfall events. In many cases aquatic organisms are exposed to mixtures of chemicals, which may lead to greater non-target risk than that predicted based on traditional risk assessments for single chemicals. We evaluated the potential toxicity of environmental mixtures of 5 classes of pesticides using concentrations from water samples collected from approximately 50 sites on midwestern streams during late spring or early summer runoff events in 1989 and 1998. Toxicity index values are calculated as the concentration of the compound in the sample divided by the EC50 or LC50 of an aquatic organism. These index values are summed within a pesticide class and for all classes to determine additive pesticide class and total pesticide toxicity indices. Toxicity index values greater than 1.0 indicate probable toxicity of a class of pesticides measured in a water sample to aquatic organisms. Results indicate that some samples had probable toxicity to duckweed and green algae, but few are suspected of having significant toxicity to bluegill sunfish or chorus frogs. PMID- 12079131 TI - Human vascular endothelial cells express pattern recognition receptors for fungal glucans which stimulates nuclear factor kappaB activation and interleukin 8 production. Winner of the Best Paper Award from the Gold Medal Forum. AB - Fungal cell wall glucans nonspecifically stimulate various aspects of innate immunity via interaction with membrane receptors on macrophages, neutrophils, and natural killer cells. We investigated the binding of water-soluble glucans in primary cultures of normal human coronary or dermal vascular endothelial cells (VECs). Membranes from VECs exhibited saturable binding. Competition studies demonstrated the presence of at least two glucan binding sites on VECs. Glucan phosphate competed for all binding sites with a KD of 3.7 microM for coronary VECs and 11 microM for dermal VECs, respectively. Laminarin, a low molecular weight glucan, competed for 47 to 51 per cent of binding (KD = 2.8-2.9 microM), indicating the presence of at least two binding sites. Glucan (1 microg/mL) stimulated VEC nuclear factor kappaB nuclear binding activity and Interleukin 8 expression--but not that of vascular endothelial growth factor--in a time dependent manner. This is the first report of pattern recognition receptors for glucan on human VECs. It also provides the first evidence that glucans can directly modulate the functional activity of VECs by stimulating cytokine gene. These results provide new insights into the mechanisms by which the host recognizes and responds to fungal cell wall products and suggests that the response to glucans may not be confined to leukocytes. PMID- 12079132 TI - Career disaffection among surgeons in the era of managed care. AB - The purpose of this paper is to assess career satisfaction of surgeons in the era of managed care. The method of this assessment is by membership survey of the Southeastern Surgical Congress and the Western Surgical Association. Members were queried about their demographic characteristics, some aspects of the business of their practices, their attitudes toward retirement, their perception of the effectiveness of organized medicine as a socioeconomic advocate, and a comparison of each surgeon's career satisfaction before and during the era of managed care. Three hundred seventy-three usable surveys were returned, computerized, and analyzed statistically. Summaries of categorical variables are presented as percentages for various groupings of the categorical set. Results reveal a broad based career disaffection in all areas queried, most notably in the business of surgical practice, attitude toward retirement and retirement alternatives, and the advocacy role of organized medicine for their issues. There is an almost unanimous desire for the American College of Surgeons to become their aggressive socioeconomic advocate. The data support a conclusion that surgeons perceive they are not represented in a realistic manner with the insurance companies and the government when their core issues are decided. PMID- 12079133 TI - The safety and efficacy of prosthetic hernia repair in clean-contaminated and contaminated wounds. AB - Prosthetic mesh reinforcement of abdominal wall hernias has gained acceptance as a result of its ease of placement and a favorably low incidence of hernia recurrence. However, its use in contaminated wounds secondary to open bowel exposure is felt to be contraindicated because of potential septic complications and lack of incorporation. The impact of permanent mesh placement in contaminated fields on wound morbidity, hernia recurrence, and mortality was examined. Records of 24 consecutive patients having permanent mesh placement in contaminated wounds for repair of abdominal wall hernias between 1994 and 2001 were reviewed. Factors examined included age, hernia type, body mass index, comorbidity, degree of contamination, concurrent gastrointestinal procedures, wound morbidity, and mortality. The mean age and body mass index were 63 years and 26.1 kg/m2 respectively. Twelve patients had risk factors for wound complications or were immunocompromised. There were 11 incisional, eight parastomal, two femoral, and two inguinal repairs and one obturator hernia repair. Twenty-three were repaired with polypropylene and one with Gore-Tex mesh. Prosthetic herniorrhaphy was performed in nine patients with ostomies already in place and in 15 patients with concomitant bowel resections. Of those with bowel resections five had enterocutaneous fistulae, three had bowel resection because of injury during mobilization, six had resections for necrotic bowel, and one had enterostomy closure. Fourteen cases were clean contaminated and ten contaminated. Eight cases were performed under emergency conditions. Wound-related morbidity occurred in five patients (21%) and in all but one was limited to cellulitis and minor wound infections. Three patients died, but in all cases death was unrelated to the surgical procedure. No patient required mesh removal. One patient had a recurrent hernia after parastomal repair. Placement of permanent mesh prostheses in clean contaminated and contaminated operative fields can be performed with minimal wound-related morbidity and patient mortality. Utilization of permanent mesh in these wounds is associated with a low incidence of hernia recurrence and eliminates the need for further surgery. PMID- 12079134 TI - Laparoscopic repair of large incisional hernias. AB - Incisional hernias after abdominal operations are a significant cause of long term morbidity and have been reported to occur in 3 to 20 per cent of laparotomy incisions. Traditional primary suture closure repair is plagued with up to a 50 per cent recurrence rate. With the introduction of prosthetic mesh repair recurrence decreased, but complications with mesh placement emerged ushering in the development of laparoscopic incisional herniorrhaphy. The records of patients who underwent laparoscopic incisional hernia repair between June 1, 1995 and September 1, 2001 were reviewed. Patient demographics, hernia defect size, recurrence, operative time, and procedure-related complications were evaluated. Fifty patients (22 male and 28 female, mean age 57 years with range of 24-83) were scheduled for laparoscopic incisional hernia repair between June 1, 1995 and September 1, 2001. The average patient was obese with a mean body mass index of 35.8 kg/m2 (range 16-57 kg/m2). Two patients (4%) had primary ventral hernias. Forty-eight patients (96%) had incisional hernias with 22 (46%) of these previously repaired with prosthetic mesh. Mean defect size was 206.1 cm2 (range 48-594 cm2). The average mesh size was 510.2 cm2 (range 224-1050 cm2). Gore-Tex DualMesh and Bard Composite Mesh were used in 84 and 16 per cent of the repairs, respectively. Mean operating time was 97 minutes. There were no deaths. Complications were seen in 12 per cent patients (six occurrences) and included two small bowel enterotomies, a symptomatic seroma requiring aspirate, a mesh reaction requiring a short course of intravenous antibiotics, and trocar site pain (two patients). There were no recurrences during a mean follow-up of 41 months (range 3-74 months). We conclude that laparoscopic incisional herniorrhaphy offers a safe and effective repair for large primary and recurrent ventral hernia with low morbidity. PMID- 12079135 TI - Small watercraft injuries in children. AB - The speed and use of small watercraft have increased dramatically in recent years. We report our experience with pediatric trauma resulting from small watercraft accidents. We conducted a retrospective chart review including all children admitted with injuries sustained in small watercraft accidents. Sixteen children were included; nine were injured in jet ski accidents and seven in accidents involving other craft. Jet ski accidents tended to result in more serious injuries (closed-head injuries, hollow and solid viscus injuries, chest trauma, spinal injuries leading to paralysis, and death) than those sustained in accidents with small boats. Skin and soft-tissue injuries and long-bone fractures were the most frequent injuries following accidents with other small boats. Six of eight children (75%) injured on jet skis required operative interventions. Only three of seven (43%) children in other watercraft accidents required surgery (P < 0.05 jet ski vs other watercraft). When compared with children injured in accidents involving small boats those involved in jet ski accidents tended to have more serious injuries and require operative intervention more frequently. A high index of suspicion for serious injuries must be maintained when evaluating children with this mechanism of injury. PMID- 12079136 TI - Postmastectomy radiation in patients with four or more positive nodes. AB - Postmastectomy radiotherapy (PMR), a local therapeutic modality, is recommended to treat breast cancer patients with multiple involved axillary lymph nodes (a marker of increased systemic risk). Bothered by this conceptually flawed treatment approach we evaluated the impact of PMR on the treatment of women with four or more involved axillary lymph nodes. We identified 1164 patients treated from 1982 through 1999 with mastectomy. We reviewed the records of the 223 who demonstrated four or more positive axillary lymph nodes. Of these 128 were treated by mastectomy only and 95 by PMR. The mastectomy-only group demonstrated a mean tumor size of 3.5 cm, a median of seven axillary nodes involved, and a median of 24.9 nodes harvested. The PMR group had a mean tumor size of 4.3 cm with nine positive nodes out of a median total of 23.3 harvested. The difference in mean tumor size was statistically significant (P = 0.01). The locoregional recurrence (10.9% vs 12.6%), distant recurrence rates (42.2% vs 35.8%), and 5 year survival (51% vs 55%) were not statistically different between the mastectomy-only group versus the PMR group, respectively. Adding PMR to breast cancer treatment demonstrated no improvement in outcome. Despite limitations of this retrospective study the results strongly support evaluation of PMR by a high quality randomized prospective trial. PMID- 12079137 TI - Laparoscopic paraesophageal hernia repair has an acceptable recurrence rate. AB - Recent literature has reported as high as a 42 per cent recurrence rate after laparoscopic paraesophageal hernia repair (LPEHR). We report long-term follow-up in a cohort of patients undergoing LPEHR at Vanderbilt University. Thirty-one patients underwent attempted LPEHR between September 1993 and May 2000. Six of 31 patients (19%) were converted to an open procedure and were excluded from the study. All patients had complete excision of the sac, primary closure of the crura, and an antireflux procedure. An Institutional Review Board-approved follow up barium esophagram was performed at a mean of 25 months postoperatively. Three experienced laparoscopic surgeons (K.S., M.H., and W.R.) collectively reviewed the esophagrams for evidence of recurrence. The mean age of patients was 61 years (range 41-92). There were six males and 19 females. Fifteen of 25 patients (60%) returned for an esophagram. Only one of 15 patients (7%) had a recurrent paraesophageal hernia. However, five of 15 patients (33%) had herniated an intact wrap 2 to 4 cm above the diaphragm. The patient with a true paraesophageal hernia recurrence returned with symptoms of dysphagia. Two of the five patients (40%) with a herniated wrap complained of heartburn, which was controlled with a proton pump inhibitor. All other patients were asymptomatic. Our recurrence rate of true paraesophageal hernias after LPEHR is low (7%) and compares with that reported in the literature for open repairs. However, 33 per cent of the patients in this study were found to have a herniated wrap. Because there is no risk of strangulation we have not operated on any of these patients. LPEHR is our procedure of choice for Type II and III hiatal hernias with good symptom relief and a low true recurrence rate. PMID- 12079138 TI - Biliary reconstruction is enhanced with a collagen-polyethylene glycol sealant. AB - Bile leaks occur in up to 27 per cent of liver transplant patients after biliary reconstruction. Synthetic sealants have not been investigated for these biliary procedures. We performed a randomized controlled study to evaluate a novel absorbable polyethylene glycol/collagen biopolymer sealant (CT3 Surgical Sealant) after incomplete end-to-end choledochocholedochostomy (CDCD) in pigs. Pigs (n = 18) underwent transection of the common bile duct and incomplete CDCD over a T tube, leaving a one-sixth circumferential defect anteriorly. Animals were randomly assigned to treatment (CDCD with sealant, n = 9) or control (no sealant, n = 9). Drains were used to monitor leak volume and bilirubin (bili) concentration. Cholangiography was performed on postoperative day 3. Leaks were defined as drain bili/serum bill > 3, total drain output > 10 mL/kg, and/or extravasation on cholangiography. Animals sacrificed at 3 and 8 weeks (n = 4 and n = 5 from each group, respectively) underwent pathologic examination of the CDCD site. Statistical methods included Student's t test, chi2, linear regression, and analysis of variance procedures. The control group had a higher drain output rate over the first 4 postoperative days than the treatment group (P < 0.05, analysis of variance). Five of nine (56%) control and one of nine (11%) treatment animals had a bile leak (P < 0.05, chi2). There was no major inflammatory response to the sealant versus controls. We conclude that CT3 is effective in decreasing biliary leaks in an incomplete CDCD porcine model with no major adverse pathologic changes. This sealant should be considered for trials for biliary reconstruction in humans. PMID- 12079140 TI - The economic impact of motor vehicle crashes: the cost of restrained versus unrestrained occupants in South Carolina. AB - The purpose of our study was to evaluate the difference in hospital charges between restrained and unrestrained motor vehicle occupants. We were also interested in identifying whether a relationship existed between restraint usage and insurance status. The data for this study were collected from two sources. The 1998 and 1999 Crash Outcome Data Evaluation System was used to compare the hospital charges and restraint usage for all motor vehicle crashes in the state of South Carolina. The patient's insurance status was also analyzed. The hospital charges and restraint usage were also compared from one of the state's Level I trauma centers for the year 1999. These data were prospectively collected and retrospectively reviewed. The average inpatient hospital charges for the state of South Carolina were approximately 25 per cent greater or $4500 more per admission for an unrestrained versus a restrained occupant. There also appears to be a relationship between payer status and restraint usage. Medicaid and self-pay patients were least likely to be using a restraining device at the time of a motor vehicle crash. The data from a Level I trauma center showed similar but more dramatic trends. Hospital charges for unrestrained occupants were 87 per cent higher than those for restrained occupants, which translates into a cost difference of $22,358 per hospital admission. This study highlights the significant economic impact of a simple preventative strategy. The cost savings for a single Level I trauma center translate into almost 9.4 million dollars for a single year. There also appears to be a relationship between usage of a restraining device and payer status. PMID- 12079139 TI - An analysis of Eastern Association for the Surgery of Trauma practice guidelines for cervical spine evaluation in a series of patients with multiple imaging techniques. AB - We conducted a retrospective review of 124 consecutive patients who received all of the following studies between October 1998 and December 1999: three-view plain films (3VPF), full CT survey (CTS), and MRI of the cervical spine. We compared the EAST guidelines for 1) patients with persistent neck pain, 2) those with neurologic deficits (NDs), and 3) those who were obtunded in our study group to determine whether EAST recommendations would risk a significant missed injury rate. The average age was 28 years (range 5 months-78 years). There were 94 males and 30 females. The mean Injury Severity Score (ISS) was 16.8 and the mean Glasgow Coma Score (GCS) 10.87. The most common mechanism of injury was motor vehicle crash (58%) followed by falling (15%), auto versus pedestrian (9%), all terrain vehicle accident (4%), assault (3%) and other (11%). For comparisons we identified a group of 33 patients with normal mental status and normal 3VPF. Twenty patients had MRI for persistent neck pain. Eleven of 20 had normal MRI. The nine abnormal MRIs showed: six ligamentous injuries, two cord compressions, and one nonligamentous soft-tissue injury. Thirteen of the 33 patients had MRI for ND. Six had normal MRI and all these NDs resolved. The remaining seven MRIs showed: two disc herniations, two cord contusions, one cord edema, one lumbar fracture, and one brachial plexus avulsion. We also examined a group of 51 obtunded patients with normal 3VPF. Thirty-six of 51 had normal CTS and MRI. Ten patients had an abnormal MRI, two an abnormal CTS, and three abnormal MRI and CTS. No obtunded patient with an adequate 3VPF had an injury identified below C2 using CTS and MRI. In the 10 patients with abnormal MRI the mean age was 28.4 years, the mean GCS 6.6 (P = 0.0025), and the mean ISS 24.3 (P = 0.03) (Wilcoxson two-sample test). The injuries identified by MRI were four disc herniations, two ligamentous injuries, two soft-tissue traumas, one meningeal tear, and one cord transection. Thirty per cent of patients with persistent neck pain had potentially unstable injuries not detected by 3VPF or CTS. Fifty-four per cent of patients with ND had abnormal MRI. Twenty-two per cent of obtunded patients with normal 3VPF and CTS had an abnormal MRI. These patients have a significantly lower GCS and a higher ISS. Six per cent of these injuries were potentially unstable. Our data support EAST guidelines for patients with persistent neck pain and ND. The guidelines for obtunded patients appear safe in detecting bony injury but may not be sensitive enough for unstable ligamentous injury and significant disc herniations. PMID- 12079141 TI - Genistein inhibits tamoxifen effects on cell proliferation and cell cycle arrest in T47D breast cancer cells. AB - Tamoxifen is an antiestrogen used in the treatment of estrogen receptor-positive breast cancer in postmenopausal women. It functions by competitively inhibiting the estrogen receptor and inducing apoptosis and G1 cell cycle arrest. Genistein is a soy phytoestrogen that inhibits breast cancer cell growth in vitro at doses of 10 microM or above. At lower doses genistein may stimulate cell growth and entry into the cell cycle. We hypothesized that treatment with low-dose genistein would reverse the inhibitory effects of tamoxifen in estrogen-receptor-positive breast cancer cells. Cell cycle kinetics and cell proliferation in T47-D human breast cancer cells were examined after exposure to genistein and tamoxifen in a low-estrogen environment designed to mimic a post-menopausal state. Cell proliferation was assessed by a colorimetric assay. Cell cycle kinetics were determined by flow cytometry. Tamoxifen caused G1 arrest and a decrease in proliferation. Genistein reversed the inhibitory effects of tamoxifen on both proliferation and G1 arrest. Thus low-dose genistein was able to inhibit the therapeutic effects of tamoxifen in this postmenopausal model of breast cancer. PMID- 12079142 TI - Inflammatory breast cancer presenting with acute central venous thrombosis: a case report. AB - Inflammatory breast cancer is a relatively rare disease associated with varied presentations, which frequently obscure the underlying diagnosis. We discuss a 54 year-old woman who presented to our emergency department with a chief complaint of right arm swelling and was admitted to the surgical service with an initial diagnosis of spontaneous brachial, axillary, and subclavian venous thrombosis. Further workup revealed an extensive inflammatory breast cancer. To our knowledge there have been no literature reports of inflammatory breast carcinoma in which the sole presenting symptom complex was due to central venous thrombosis. PMID- 12079143 TI - Preoperative anemia in colon cancer: assessment of risk factors. AB - Anemia is common in cancer patients and is associated with reduced survival. Recent studies document that treatment of anemia with blood transfusion in cancer patients is associated with increased infection risk, tumor recurrence, and mortality. We therefore investigated the incidence of preoperative anemia in colorectal cancer and assessed risk factors for anemia. Prospective data were collected on 311 patients diagnosed with colorectal cancer over a 6-year period from 1994 through 1999. Patients were stratified by age, gender, presenting complaint, preoperative hematocrit, American Joint Committee on Cancer (AJCC) stage, and TNM classification. Discrete variables were compared using Pearson's Chi-square analysis. Continuous variables were compared using Student's t test. Differences were considered significant when P < 0.05. The mean age of the study cohort was 67 +/- 9.2 with 98 per cent of the study population being male. The mean AJCC stage was 2.2 +/- 1.2 and the mean preoperative hematocrit was 35 +/- 7.9 with an incidence of 46.1 per cent. The most common presenting complaints were hematochezia (n = 59), anemia (n = 51), heme-occult-positive stool (n = 33), bowel obstruction (n = 26), abdominal pain (n = 21), and palpable mass (n = 13). Preoperative anemia was most common in patients with right colon cancer with an incidence of 57.6 per cent followed by left colon cancer (42.2%) and rectal cancer (29.8%). Patients with right colon cancer had significantly lower preoperative hematocrits compared with left colon cancer (33 +/- 8.5 vs 36 +/- 7.4; P < 0.01) and rectal cancer (33 +/- 8.5 vs 38 +/- 6.0; P < 0.0001). Patients with right colon cancer also had significantly increased stage at presentation compared with left colon cancer (2.3 +/- 1.3 vs 2.1 +/- 1.2; P < 0.02). Age was not a significant risk factor for preoperative anemia in colorectal cancer. We conclude that there is a high incidence of anemia in patients with colon cancer. Patients with right colon cancer had significantly lower preoperative hematocrits and higher stage of cancer at diagnosis. Complete colon evaluation with colonoscopy is warranted in patients with anemia to improve earlier diagnosis of right colon cancer. A clinical trial of preoperative treatment of anemic colorectal cancer patients with recombinant human erythropoietin is warranted. PMID- 12079144 TI - Bilateral giant adrenal myelolipomas: a case report. AB - Myelolipoma is a relatively rare benign tumor composed of fatty tissue and bone marrow elements. It is frequently associated with the adrenal glands but may exist as a solitary mass elsewhere. Adrenal myelolipomas are typically nonfunctioning and asymptomatic. They may be associated with an endocrine disorder such as Cushing's disease, Addison's disease, or hyperaldosteronism; however they are most often discovered incidentally. Their size is usually less than 5 cm and they are managed nonoperatively. We report a case of bilateral giant adrenal myelolipoma producing abdominal pain in a 54-year-old man, who presented to his primary care physician with complaints of right shoulder pain with vague abdominal discomfort. Imaging studies identified bilateral suprarenal masses measuring 12 x 14 cm on the right and 8 x 10 cm on the left. These masses were consistent with fatty tissue radiographically. In addition a focal 5 x 4-cm mass was identified in the transverse colon. Because the patient was symptomatic and a diagnosis of liposarcoma could not be excluded he was taken to the operating room for exploratory laparotomy with excision of the masses and a transverse colectomy. Final histologic analysis identified bilateral adrenal myelolipomas and a solitary lipoma of the transverse colon. His postoperative course was uneventful with relief of the pain. Despite its benign nature and rare growth beyond 5 cm myelolipoma of the adrenal gland is best managed with excision in the symptomatic patient. Preservation of adrenal tissue is vital so as not to commit patients to a lifetime of steroid replacement. PMID- 12079145 TI - The incidence and significance of free air after percutaneous endoscopic gastrostomy. AB - Percutaneous endoscopic gastrostomy (PEG) is well established as a safe and effective means of providing enteral feeding access in patients unable to tolerate oral feeding. There is some question, however, as to the true incidence of free air after PEG and the clinical significance of free air in these patients. We report our experience with 119 patients over 4 years who underwent placement of a percutaneous gastrostomy tube. This study is a retrospective review of percutaneous endoscopic gastrostomies performed by the Critical Care Service for Surgery (CCSS). A database of percutaneous endoscopic gastrostomies performed by the CCSS was maintained from September 1997 through December 2001. Complications of percutaneous gastrostomies were added to the database when noted. The electronic medical record of all patients was reviewed for the results of radiographic studies. Prior abdominal operations were noted as well as gastrostomy tube complications and outcome. A total of 115 intensive care unit patients underwent PEG placement by the CCSS. This total includes 18 patients who had undergone prior upper abdominal surgery, Three additional patients who underwent placement of a gastrostomy tube by vascular interventional radiology and one patient who underwent PEG placement by the ear, nose, and throat service were brought to the attention of CCSS secondary to complications for a total of 119 patients. Only four patients (3.4%) were found to have free air on subsequent chest radiograph. Six patients (5.2%) were found to have free air on abdominal CT scans. Two patients with free air on CT underwent exploratory celiotomy as a result of additional signs of peritonitis. Both were negative explorations. The incidence of free air after PEG in our experience is significantly less than the incidence in previous studies. In patients with free air after PEG placement exploratory celiotomy is not indicated in the absence of other clinical findings of peritonitis. Additionally it was noted that PEG placement could safely be performed in patients with prior upper abdominal surgery with a low incidence of complications. PMID- 12079146 TI - There is no difference in survival between African-American and white women with breast cancer. AB - A difference in survival by race in women with breast cancer has been reported. We examined survival of African-American (AA) and white (W) women diagnosed with breast cancer and evaluated variables that may affect survival at a university teaching hospital. A retrospective review of tumor registry data for breast cancer was performed in all AA and W female breast cancer patients diagnosed between the years 1990 and 1999 at a university hospital. Survival between AA and W women diagnosed with breast cancer during that period was compared using Kaplan Meier analysis. Variables including age and stage of disease at diagnosis, receptor status, treatment, and tobacco exposure were evaluated utilizing Chi square testing. A P value <0.05 was considered statistically significant. A total of 585 AA and W women were diagnosed with breast cancers between the years 1990 and 1999. Mean ages were 51.8 years for AA and 56.9 years for W (P = 0.001). Overall survival (7.8 years AA and 7.6 years W) and survival by stage were not statistically different between the groups. AA patients were younger (P = 0.001), presented with higher-stage tumors (P = 0.017), more often had positive axillary lymph nodes (P = 0.012), more often were estrogen and progesterone receptor negative (P = 0.004), and more often were premenopausal (P = 0.048). AA women were more likely treated with chemotherapy while W women tended to receive hormone therapy (P = 0.01). Statistical significance was not reached for differences in tumor histology or tobacco exposure. We conclude that despite presenting with worse prognostic indicators AA women experience survival equivalent to their W counterparts. PMID- 12079147 TI - Medicare payment system for hospital inpatients: diagnosis-related groups. AB - Diagnosis-Related Groups (DRGs) are categories of patient conditions that demonstrate similar levels of hospital resources required to treat the conditions. Each inpatient that is discharged from an acute care hospital can be classified into one of the 506 DRGs currently utilized by the Medicare program. The Medicare DRG prospective payment methodology has been in use for almost two decades and is used by hospital managers for planning and decisionmaking. The viability of DRGs for future prospective payment depends on the ability to keep up with the times through updates of the current methodology. PMID- 12079148 TI - Medicare's risk-adjusted capitation method. AB - Since 1997, the method to establish capitation rates for Medicare beneficiaries who are members of risk-bearing managed care plans has undergone several important developments. This includes the factoring of beneficiary health status into the rate-setting calculations. These changes were expected to increase the number of participating health plans, accelerate Medicare enrollment growth, and slice Medicare spending. PMID- 12079149 TI - Medicare's fee schedule for hospital outpatient care. AB - Medicare's hospital outpatient prospective payment system (OPPS) went live on August 1, 2000, after a decade of developmental work. The new system introduced a fee schedule that replaced the cost-related methods that Medicare previously used to reimburse various hospital outpatient services. Hospitals are now paid predetermined rates or fees based on the Ambulatory Patient Classification (APC) groups assigned to the services that Medicare patients receive during outpatient encounters. The new system aims to simplify Medicare's intricate cost-based reimbursement policies, improve hospital efficiency, ensure that payments are sufficient to compensate hospitals for reasonable Medicare costs, and reduce Medicare coinsurance amounts for beneficiaries. Implementation of OPPS-related administrative and operational changes has been a major challenge for hospitals. PMID- 12079150 TI - Inpatient rehabilitation facilities are now paid prospective rates. AB - On January 1, 2002, Medicare began to replace its cost-related inpatient reimbursement method for rehabilitation hospitals and rehabilitation units in hospitals with a prospective payment system. Under the new system, Medicare pays a predetermined, fixed amount per discharge, depending on the patient's impairment level, functional status, comorbid conditions, and age. Reduced or additional amounts are paid for early transfers, short-stay outliers, patients who expire before transfer, and cost outliers. The overall objective of the new case-based system is to provide incentives for rehabilitation facilities to furnish intensive inpatient services efficiently without tarnishing the quality of care or constraining access to care. Federal actuaries estimate that the new system will cost Medicare an additional $70 million between January 1, 2002, and September 30, 2003. The actual increase may be larger. PMID- 12079151 TI - Prospective per diem rates for skilled nursing care. AB - On July 1, 1998, Medicare's cost-related reimbursement method for skilled nursing facility care was replaced with a prospective payment system that includes a case mix adjustment based on the Resource Utilization Groups to which Medicare residents are assigned. Shortly thereafter, Congress modified the new system in response to the industry's complaints about low payment rates. The new system aims to align Medicare payments more closely with the costs facility's incur in serving Medicare residents and slow Medicare spending growth. Recent rate increases have reduced the new system's ability to trim Medicare outlays. PMID- 12079153 TI - Medicare payment system for ambulatory surgical centers. AB - Ambulatory Surgical Centers are paid under a fee schedule for eight service cost categories. The fee schedule is geographically wage adjusted and is updated annually. Anticipated future adjustments to the Ambulatory Surgical Centers prospective payment system (PPS) includes conversion to resource based rates. PMID- 12079152 TI - Prospective rates for episodes of home health care. AB - Starting October 1, 2000, Medicare began paying providers of home health care at fixed, predetermined rates for services and items bundled into 60-day episodes of home health care. The episode payment rates vary with the patient's clinical, functional, and services utilization characteristics. This new approach was expected to redistribute Medicare payments among home health agencies, extend the cost savings introduced by the Interim Payment System, improve the coordination of services, and reduce the number of unnecessary home health visits. PMID- 12079154 TI - Medicare fees for physician services are resource-based. AB - Beginning January 1, 1992, Medicare has relied on a resource-based relative value scale (RBRVS) to establish physician fees. Medicare pays 80 percent of the lower of the amount a physician bills for the service or the fee schedule amount. The patient is responsible for the remaining 20 percent, as well as the annual Part B deductible of $100, plus any additional amount the physician may be allowed to bill. Rarely is the billed amount below Medicare's fee schedule amount. Adoption of the RBRVS fee schedule severed the link between the amount a physician charged for a service and the amount Medicare paid for it. RBRVS implementation required significant changes in the coding system used to document and bill physician services, particularly medical visits and consultations. PMID- 12079156 TI - Conditions before collapse of the contralateral hip in osteonecrosis of the femoral head. AB - BACKGROUND: Osteonecrosis of the femoral head is a disease which may cause rapid femoral head collapse in relatively young patients. Clinical results of prosthesis replacement in osteonecrosis patients are generally poorer than in age matched patients with other diagnoses. The ultimate goal in treating these patients is therefore to preserve not replace the involved femoral head. The purposes of this study were to analyze outcomes by examining osteonecrotic hips before collapse and to justify the role of early surgical intervention for osteonecrosis of the femoral head. METHODS: Between 1993 and 1997, there were 275 consecutive patients with osteonecrosis of the femoral head who received joint replacement on 1 side. Among these patients, 100 contralateral hips with evidence of osteonecrosis in the stage before collapse were included in this study. Demographic information, laboratory data, and radiographs were reviewed and compared between patients whose collapse time was less than 1 year, between 1 and 2 years, and more than 2 years, and those who had no collapse at the latest follow-up. RESULTS: At the final follow-up, the overall collapse rate was 78%, and the collapse time was generally within 2 years. Patients with younger age (less than 50 years old), higher activity levels, and higher serum triglyceride levels were at higher risk of rapid collapse. CONCLUSION: In conclusion, we found that the disease progression rate is high in patients who have bilateral femoral head involvement. Early surgical intervention to preserve the femoral head should be adopted to prevent early femoral head collapse in these patients. In addition, risk factors such as high activity levels and abnormal lipid metabolism should also possibly be addressed in the therapeutic modality. PMID- 12079155 TI - Resectable gastric cancer: operative mortality and survival analysis. AB - BACKGROUND: This study evaluated the survival outcome and determined the prognostic factors for gastric cancer patients who underwent gastric resection in the past 6 years. METHODS: Between 1994 and 2000, a total of 1,322 patients with gastric cancer who underwent gastric resection in our hospital comprised the study subjects. Their mean age was 61.1 (range, 14-92) years. There were 865 male and 457 female patients. Total gastrectomy was performed in 389 (29.4%) and distal gastrectomy in 933 patients. Curative resection was performed in 961, and palliative resection in 361 patients. A D2 or greater lymphadenectomy was required for curative resection. Patients received postoperative chemotherapy if they underwent palliative resection. RESULTS: Early or pT1 gastric cancer accounted for 17.7% and lymph node metastasis for 62.1% of all resected cases. The overall operative mortality and morbidity rates were 3.3% and 18.0%, respectively. The operative mortality for palliative total gastrectomy was particularly high (8.5%). The overall cumulative 5-year survival rate of all resected patients was 45.6%, and it was 57.0% after curative resection. Multivariate analysis revealed that lymph node metastasis, serosal invasion, peritoneal seeding, positive resection margin, liver metastasis, old age, tumor size, and lymphatic invasion were independent prognostic factors. CONCLUSION: The most important prognostic factors for survival were lymph node metastasis, serosal invasion, peritoneal seeding, positive resection margin, liver metastasis, old age, tumor size, and lymphatic invasion. The operative mortality and survival outcome of our gastric cancer patients after gastric resection compared favorably with those of other series in other countries. PMID- 12079157 TI - High false-positive rate of cytokeratin-19 in detecting circulating tumor cells for nasopharyngeal carcinoma. AB - BACKGROUND: Nasopharyngeal carcinoma (NPC) harbors a higher metastatic potential than other head and neck cancers. In order to seek a possible surrogate marker for early detection of recurrent or metastatic disease, we tested the feasibility of cytokeratin-19 (CK-19)-nested reverse-transcription polymerase chain reaction (RT-PCR) for detecting circulating tumor cells in NPC patients. METHODS: Two tubes of blood were sequentially collected in individual draws from 7 NPC patients and 15 healthy persons. Total ribonucleic acid (RNA) was extracted from blood cells and treated with deoxyribonuclease. The RNA was then subjected to RT and nested PCR with specific CK-19 primers. The reaction products were run on an agarose gel and visualized under UV light. The sequences of the products were determined using an ABI377 automatic sequencer. RESULTS: Among the 7 NPC cases, 4 cases presented CK- 19 expression with 2 in both tubes, 1 in the first tube, and 1 in the second tube. In the control group, 8 of 15 cases also presented CK-19 expression with 6 in both tubes and 2 in the second tube resulting in a 53.3% false-positive rate. Incidentally, an aberrant splicing product lacking exon 4 of CK-19 messenger RNA was discovered. CONCLUSION: Results of the present study indicate that the CK-19-nested RT-PCR is not suitable for detecting circulating tumor cells in NPC patients because of a high false-positive rate in the control group. The reason for the high rate of false-positives may be attributed to pseudogenes, different blood cell separation methods, or illegitimate expression of CK-19 in blood cells. PMID- 12079158 TI - Early surgical management for heterotopic ossification about the elbow presenting as limited range of motion associated with ulnar neuropathy. AB - BACKGROUND: The formation of heterotopic ossification (HO) about the elbow after traumatic injury has been well documented in the literature. The optimal treatment, however, for ectopic bone associated with restricted range of motion and ulnar nerve entrapment syndrome has not been established. METHODS: Seven elbows with HO in 7 patients admitted to Chang Gung Memorial Hospital from April 1998 to January 1999 presented with limited range of motion and associated ulnar nerve neuropathy. All of these patients received early surgical excision of HO combined with release of the encased ulnar nerve and anterior transposition, followed by early gentle passive physical therapy and active exercise within the pain-free range of motion postoperatively. RESULTS: Almost full range of motion and complete functional ability following surgery were recovered in 6 of the 7 patients, while I patient who suffered from multiple traumatic injuries had limited improvement from 45 degrees ankylosis to 10 degrees approximately 90 degrees of a flexion-extension motion arc. CONCLUSION: Our results suggest that early surgical management combined with gentle physical therapy postoperatively is a feasible modality for treating patients with post-traumatic HO about the elbow presenting as limited range of motion and associated ulnar nerve compression syndrome. PMID- 12079159 TI - Surgical outcomes of short-segment fixation for thoracolumbar fracture dislocation. AB - BACKGROUND: Currently long-segment pedicle instrumentation for a thoracolumbar (T L) fracture-dislocation is gaining in popularity. Otherwise, short-segment fixation may be chosen as an another treatment method. This article evaluates the efficacy and complications of short-segment fixation for the treatment of thoracic or lumbar spine fracture-dislocation. METHODS: Twenty patients with thoracic or lumbar spine fracture-dislocation were included in the study. The mean follow-up period was 3 years. Clinical, neurologic, radiologic (angle of deformity, displacement percentage, fusion rate), and complication outcomes were analyzed retrospectively. RESULTS: The rate of failure, defined as an increase of 10" or more in local kyphosis, an increase of 10% or more in displacement percentage, the development of pseudarthrosis, and/or implant failure, was analyzed. Short-segment instrumentation had a higher failure rate in the lower lumbar region (L3-L5) and a lower rate at the thoracolumbar junction (T11-L2). CONCLUSION: Thoracolumbar fracture-dislocation required firmer fixation especially in the low lumbar region, and short-segment fixation resulted in a high failure rate. Circumferential anterior and posterior fusion often played a role in certain severely injured cases. Good postoperative spinal alignment is crucial to a good outcome. PMID- 12079160 TI - Double partial monosomies (10p- and Xp-) in a female baby with choanal atresia. AB - Chromosomal abnormalities involving double partial monosomies are very rare. A female infant with non-mosaic monosomy 10p13-->10pter along with monosomy Xp11.4- >Xpter which arose de novo is described. The clinical manifestations of this patient included microcephaly, mild synophrys, short and down-slanted palpebral fissures, ptosis of the left eye, long eyelashes, a depressed nasal bridge, dysplastic ears, micrognathia, a short neck. sensorineural hearing impairment, and severe growth retardation. Left choanal atresia and laryngomalacia were detected by flexible fibroscopy. No signs of hypoparathyroidism or defective cellular immunity could be found. Fluorescence in situ hybridization (FISH) with whole-chromosome painting probes for chromosomes 10 and X was performed, which excluded the possibility of cryptic translocations of the involved chromosome segments. No submicroscopic chromosome 22q11 deletion could be found by FISH. Thus this very rare coexistence of double independent partial monosomies was confirmed. There are no previous reports of such concurrent double partial monosomies. PMID- 12079161 TI - Oral wooden stick injury complicated by meningitis and brain abscess. AB - Meningitis is rarely seen following oral injury. We describe a 3-year-old boy developing meningitis and brain abscess following a penetrating oral wooden stick injury. There was no cerebrospinal fluid rhinorrhea noted. A cerebrospinal fluid culture yielded viridans streptococcus. Brain magnetic resonance imaging and computed tomography revealed a multiloculated ring-enhancing mass. This patient underwent surgical drainage and completed 8-week antibiotic therapy. The patient demonstrated a late and dismal complication of a penetrating oral injury. At 2 year follow-up the patient was in good condition. A penetrating oral wooden stick inury should be regarded as potentially serious. PMID- 12079162 TI - Leiomyomatosis of mesenteric lymph nodes associated with duodenal adenocarcinoma. AB - Leiomyomatosis of lymph nodes is an extremely rare disease. Only a few cases have previously been reported in pelvic lymph nodes. They were related to a benign uterine leiomyoma, a metastasizing uterine leiomyoma, an endometrial adenocarcinoma, and an ovarian endometrioid carcinoma. We report on a case of leiomyomatosis of the mesenteric lymph nodes associated with a duodenal adenocarcinoma with no history of uterine leiomyoma or any gynecological malignancy. The patient, a 56-year-old woman, was found to have an adenocarcinoma of the duodenum. All mesenteric lymph nodes removed showed leiomyomatosis, which was verified by immunohistochemical study showing positive immunostaining for smooth muscle actin, desmin, and vimentin, but negative staining for HMB-45. It is necessary to make a differential diagnosis from other examples of spindle cell proliferation involving lymph nodes such as a hemorrhagic spindle cell tumor with amianthoid fibers (palisade myofibroblastoma), angiomyolipoma, lymphangiomyomatosis, inflammatory pseudotumor, and Kaposi's sarcoma. PMID- 12079163 TI - A huge epiglottic cyst causing airway obstruction in an adult. AB - An epiglottic cyst causing airway obstruction is rare in an adult. Early definitive diagnosis and management obviate an unnecessary tracheostomy. We report a case of a 64-year-old woman who arrived at our hospital with progressive stridor and foreign body sensation when swallowing for 6 weeks. A hot potato voice and biphasic stridor were remarkable upon physical examination. Indirect mirror and fibroscopic examination revealed a huge epiglottic cyst. The neck lateral X-ray and computed tomography scan demonstrated a huge cystic mass over the epiglottis. A 2.5 x 3.0 cm cystic mass was removed with endoscopic CO2 laser after needle decompression. The patient was discharged on the third day after surgery without complications. An epiglottic cyst in an adult seldom causes upper airway obstruction and is easily ignored by clinicians. We emphasize that complete airway evaluation including routine check-up of the larynx is mandatory for patients with intractable obstructive airway disease. Endoscopic laser surgery is effective in the surgical removal of an epiglottic cyst. PMID- 12079164 TI - Squamous cell carcinoma arising in an epidermal inclusion cyst. AB - An epidermal inclusion cyst is a widespread benign intradermal lesion and may occur anywhere in the body. Normally, it appears as a non-tender, soft mass of variable size. Dissection usually reveals grayish-white or whitish gelatinous materials and a smooth inner surface. The overlying skin almost always shows unremarkable changes. On occasion, the cyst may rupture and induce an inflammatory reaction. It rarely turns malignant or displays a firmer mass. This study reports on a rare case of squamous cell carcinoma arising from the lining cells of an epidermal inclusion cyst, which was located in the left axillary region of a 68-year-old male patient. Clinically, it is difficult to differentiate between a benign and malignant cystic lesion. Histological examination normally yields the diagnosis. Once a diagnosis is confirmed, the tumor should be widely excised with a free margin. The outcome is always excellent. We therefore emphasize that all resected skin cystic specimens should undergo further microscopic examination to avoid any unnecessary misdiagnosis. PMID- 12079165 TI - Gun violence and children: factors related to exposure and trauma. AB - The study discussed in this article investigated the relationship between access to firearms and parental monitoring on rural youths' exposure to gun violence and examined the effect of gun violence exposure on the mental health of these youths. A survey was administered to rural students who participated in a student assistance program (n = 162) that provided in-school support groups for students in grades 6 through 12. Results indicated that a substantial number of students were exposed to gun violence and exposure was significantly related to firearm access and parental monitoring. Furthermore, gun violence exposure was significantly associated with trauma among the youths. Implications for social workers include advising high-risk clients and their families on gun removal and safe storage practices. PMID- 12079166 TI - A trauma education workshop on posttraumatic stress. AB - Adapting successfully in life involves successful management of stress. Posttraumatic stress presents a significant challenge for successful adaptation, one that is often misunderstood, devalued, dismissed, or denied. This article presents a brief and potentially effective method of group education about posttraumatic stress for "victims" of posttraumatic stress, significant others, and health worker. Appropriate use of group education for posttraumatic stress, it is argued, offers empowering meaning and purpose, and faith and hope for healing and well-being. PMID- 12079167 TI - Health implications of animal hoarding. AB - Animal hoarding is a poorly understood phenomenon, the public health implications of which are not well documented. In this study, professionals dealing with hoarding cases submitted 71 case report forms. The hoarders' residences were characterized by extreme clutter and poor sanitation that impaired ability to maintain functional households. Appliances and utilities were frequently nonfunctional, and animal excrement sometimes accumulated to the extent that the homes were unfit for human habitation. The majority of cases satisfied criteria for adult self-neglect, and dependent elderly people, children, or disabled individuals were present in many of the residences. Animal hoarding may be a sentinel for a range of medical, social, and economic problems. More research addressing the causes and features of animal hoarding is needed to shed light on appropriate interventions. PMID- 12079168 TI - Shyness and social phobia: a social work perspective on a problem in living. AB - Social phobia is classified as an anxiety disorder in psychiatric nomenclature. It represents a fear of performance or social interaction that significantly interferes with a person's social or occupational functioning. The author takes issue with the fact that social phobia is considered by many professionals to be a mental illness that is often treated best with medication. Social phobia can be conceptualized from a social work perspective as an extreme shyness that can be overcome with cognitive learning and behavioral rehearsal. This article reviews the biopsychosocial causes of social phobia and presents a summary of cognitive and behavioral interventions with empirically demonstrated effectiveness. PMID- 12079169 TI - Parents of mentally ill adult children living at home: rewards of caregiving. AB - In the context of parents caring at home for an adult child with mental illness, this study explored the positive aspects and rewards of caregiving. Specifically, it measured the extent to which parents perceive their ill child as providing assistance and support--practical and emotional--and perceive their own caregiving as emotionally and mentally rewarding. One parent from each of 93 households completed a self-administered questionnaire. All the parents reported receiving help and support from their child, but perceived the satisfaction gained from fulfilling their parental duties and from learning about themselves as far more important. Their assessment of this satisfaction was entirely unaffected by the subjective and objective burdens on them and the severity of the child's illness. PMID- 12079170 TI - Clinically sensitive peer-assisted mediation in mental health settings. PMID- 12079171 TI - Promoting community awareness to enhance mental health. PMID- 12079172 TI - Mental health recovery paradigm: implications for social work. AB - Social workers have long been involved in developing, administering, and providing services for people with psychiatric disabilities. Critics of the system, including social workers and mental health consumer-survivor practitioners, have noted that the medical model has been a driving force in policy and services provision. This model is detrimental to consumers' self efficacy and sense of hope and conflicts with a number of central social work values. The article argues that the values and beliefs of the consumer-survivor recovery movement are closely aligned with those of the profession, and that the movement offers social workers a more promising perspective from which to practice. The primary concepts and values of the evolving recovery paradigm are delineated, and implications for direct practice, administration, policy making, education, and research are discussed. PMID- 12079173 TI - Dimensions and predictions of professional involvement in self-help groups: a view from within. AB - This article focuses on an analysis of how members of self-help groups perceived professional involvement and what they defined as the dimensions of such involvement. Models to predict these identified dimensions are also suggested. The instrument developed for the study consisted of two parts: (1) demographic question and (2) 12 statements concerning self-help group members' attitudes toward professional involvement. Based on responses from 183 members of self-help groups, three categories were identfied according to their main focus: groups dealing with health issues, groups dealing with alternative lifestyles, and groups based on the 12-step model. Analysis yielded two conceptually different constructs influencing group members' attitudes: collaboration and duality. PMID- 12079174 TI - A tribute to nurses and Florence Nightingale. PMID- 12079175 TI - Asthma management: Part 1: An overview of the problem and current trends. AB - Death rates from asthma have increased or remained stable over the past decade despite increased knowledge about the pathophysiology and improved treatment of the disease, a fact that is both puzzling and disconcerting. Some children with asthma experience severe and life-interfering exacerbations separated by long periods of normal lung function and no symptoms. Asthma management plans are developed by primary care providers and the family. School nurses implement and monitor the child's response to the plan. Therefore, the school nurse needs current information about asthma management. Part I of this two-part series describes the pathophysiology of asthma and the types, risk factors, and current trends in management of the disease. The role of the school nurse in asthma management is outlined, including how he or she can influence environmental factors that precipitate asthma symptoms or exacerbations. Part II will discuss the role of the school nurse in pharmacologic management of asthma. Complementary alternative medicine for asthma management will be described, as well as health teaching for the child with asthma and their family. PMID- 12079176 TI - Are school nurses using the recommendations of the Diabetes Control and Complications Trial in the care of students with diabetes? AB - Diabetes is one of the most common chronic diseases of childhood. It is listed as a disabling condition under the Americans with Disabilities Act, and therefore accommodations must be made for these children in school. School nurses are responsible for large numbers of children, usually in more than one setting. The Diabetes Control and Complications Trial examined intensive therapy and its effects on the long-term morbidity and quality of life of diabetic adolescents. A number of recommendations were made that apply to school nurses. The purpose of this article is to determine if any studies have been conducted to explore whether school nurses are using the results of the Diabetes Control and Complications Trial to write individualized healthcare plans defining the care of students with diabetes. PMID- 12079177 TI - Type 1 diabetes in children and adolescents: what's new? AB - As providers of care for children and adolescents with Type 1 diabetes mellitus (DM), it is imperative that the school nurse is familiar both with the legal framework guiding protection of these students' rights in school and with ongoing developments regarding management of this disorder. Federal legislation, designed to prevent discrimination and to allow children and adolescents with certain disabilities and chronic disorders such as Type 1 DM to fully participate in activities within the academic setting, is a major consideration for those providing care in schools. Understanding of recent developments and future trends in the management of Type 1 DM is essential for the school nurse to facilitate optimal care for students with this disorder. These consist of new types and activity of insulin preparations, alternative devices for insulin delivery, improvements in glucose monitoring, and changes in nutritional management along with upcoming developments in Type 1 DM management. PMID- 12079178 TI - Nursing, food service, and the child with diabetes. AB - The safety and well-being of children while they are at school or day care is a major concern in our society. It is therefore important that the professionals who are entrusted with the care of young people possess the knowledge and skills necessary to provide a safe and caring environment for all children, including those with chronic disorders such as diabetes. Each day 35 children in the United States are diagnosed with diabetes mellitus. To make their school experiences positive, it is imperative that school employees be aware of the potential challenges associated with diabetes and how to meet these challenges successfully. This article discusses the role of the school nurse in forming an interdisciplinary team to work with a child with diabetes. It addresses the school nurse's role in diabetes education of food service personnel in the school. Confidentiality issues are also addressed. PMID- 12079179 TI - Research priorities in school nursing: a Delphi process. AB - As a follow-up to an Invitational Summit Conference in 1999, 27 nurse leaders representing school nursing and nursing research in child health were requested to participate in a Delphi study for purposes of prioritizing research issues in school nursing. The issues to be prioritized were identified during the Invitational Summit meeting. A three-round Delphi process was used to arrive at consensus on the most important research issues affecting school nurses today. Criteria for the rankings included policy support for the research, ability to be funded, magnitude and severity of the problem, likelihood of making a difference, and potential for partnership. The overall response rate for participation in all three rounds was 61%. Consensus was reached on the top-10 most important research issues. The two most important were consistent through all three rounds. These were (a) the impact of school nurse services on student health, and (b) the relationship between school nurse practice and educational outcomes. PMID- 12079180 TI - Positive health practices of urban minority adolescents. AB - What influences the health practices of adolescents? This study attempted to answer this question by examining factors related to health practices of urban minority adolescents. The convenience sample consisted of 65 minority adolescents who resided in an urban area. Positive correlations were found between social support and positive health practices and hope and positive health practices. However, no significant relationship was found between self-esteem and positive health practices. School nurses work closely with adolescents and are in the position to initiate and promote programs that will influence the health behaviors of minority adolescents. Suggestions are provided that will assist school nurses to meet their goal of promoting positive health practices among adolescents. PMID- 12079181 TI - Decision support tools aid school nursing practice. AB - The purpose of this article is to explain the use of technology as a tool to aid the school nurse's decision-making process. This article demonstrates how decision support tools improve school nurses' decision making regarding client care and nursing practice. Various decision support systems are discussed, examples provided, and the underlying technology explained. A listing of decision support systems and specific applications for school nurses are reviewed. PMID- 12079182 TI - Bring your first aid kit: an unannounced mock drill. AB - Disaster preparedness has taken on new significance in our country since September 11. School nurses, advocates for school safety, must address new challenges in crisis management and emergency response. Our nation's schools remain relatively safe places, yet well-known events in the last few years dictate the need for movement from attitudes of complacency and denial toward vigilance. Natural disasters, accidents, and violence can threaten the well-being and lives of students and staff, and in a few short minutes a peaceful learning environment can change into one of chaos with multiple casualties. Although schoolwide drills for events such as tornadoes, explosions, and shootings remain imperative, they do little to prepare the school nurse for her role in immediate response. Staging an unannounced mock disaster at a districtwide nurse meeting is one way to ensure a higher level of preparedness. It also acknowledges the legitimate concerns of crisis competency among school nurses who are often and understandably the most trusted first responders to health crises on campus. PMID- 12079184 TI - Setting specifications for potency assays--basic principles. AB - To develop any biological therapeutic successfully, it is necessary to characterise the product thoroughly, both physicochemically and biologically. To ensure that consistency of production is maintained, some level of control is required to limit the variability of the product from batch to batch. This premise forms the basis of specification setting. The biological activity or potency of the product must be appropriately assessed through a functional assay unless specifically justified otherwise e.g. for binding proteins where binding has been correlated with biological activity. Biological assays are particularly prone to assay variability and therefore it is necessary to design and execute the assay to reduce variability as much as possible while providing a statistically valid measure of the reproducibility of potency estimates. Therefore, specifications for potency should only be derived following establishment of a well validated and controlled bioassay and should include some measure of the variability of the estimate in addition to the actual potency estimate itself. The limits applied to those specifications need to reflect the true batch to batch consistency of the product, its nature and toxicity as well as its intended use. PMID- 12079183 TI - Consent and confidentiality issues among school-age children and adolescents. AB - This article supports the moral viewpoint that minors with decision-making capacity, regardless of their age, should be involved in their health care decisions. When clinical care proceeds without attention to a child's emotional and developmental potential, the needs of a young person to be socialized to take care of his or her health care from an early age are not met. This is ethically irresponsible. Developing competence for informed consent decisions is associated with cognitive capacity. Explicit federal guidelines and laws exist for the inclusion of minors in research. By contrast, minors are often left out of participating in the health care decisions that may affect them for a lifetime in illness and wellness preservation. For day-to-day care, respect for children as increasingly autonomous individuals, members of families, and society is derived from the ethical principle of autonomy. The school nurse is often in a unique position to promote this professional behavior because most interactions are directly with the minor and without parental presence. In addition, the health office is a "safe" place where students can go for a variety of concerns. The school nurse must be concerned about ethical practice while staying within the limits of state and federal laws regarding minors' consent to treatment and confidentiality. PMID- 12079185 TI - Strip-plot designs, mixed models, and comparisons between linear and non-linear models for microtitre plate bioassays. AB - By paying careful attention to the experimental units and, randomizing where it is reasonable, we are able to devise designs for cell culture bioassay which are statistically legitimate and practical in the laboratory. These designs, strip plot layouts, can accommodate linear or non-linear models, fixed or mixed models, and can provide good protection from location effects. Non-linear mixed models using these designs can be extended to address serial dilution error. A statistically based approach which includes randomization and proper design is a powerful tool for identifying subtle effects of factors within and outside bioassays; this attention to statistical detail is a necessary first step towards continuous improvement of biological assays. PMID- 12079186 TI - Statistical approaches to specification setting with application to bioassay. AB - Statistics has a role to play in the establishment of specification limits. This is particularly true in bioassays where the levels of uncertainty in the process and assay can be substantial. The use of statistical approaches can aid manufacturers in setting meaningful specifications and help to evaluate the ability of their processes to meet specifications long-term. PMID- 12079187 TI - International specification setting--EP perspectives. European Pharmacopoeia. AB - The regulatory framework for medicines in Europe is outlined and the role and policies of the European Pharmacopoeia in setting requirements for biotechnology products are discussed. PMID- 12079189 TI - Characterization and implications of bioassay variation. AB - Statistical models for variance components can be used to characterize the components of variance in potency estimates obtained from a bioassay. We outline these methods and discuss the implications for replication strategy. An example is provided. PMID- 12079188 TI - Validation of biological assays. PMID- 12079190 TI - Statistical considerations in the design and analysis of parallel line assays. AB - Underlying the design of any assay and further, interpretation of the results, are multitudes of assumptions and implications, ranging from 'biological' assumptions about the nature of the assay system and its response to the materials assayed to 'statistical' assumptions about the form of the dose response relationship and the distribution of the response data. As far as possible, assays should be designed and analysis carried out to assess these assumptions. Implications for the individual assay are discussed, since this is where all studies of assays necessarily begin. Consideration is then extended to implications for the combination of data and results of several assays. PMID- 12079191 TI - Selected topics in parallel line bioassay. PMID- 12079192 TI - Parallel line bioassay in practice. Practical aspects of design, analysis, and regulatory requirements from the technician's point of view. AB - Bioassay technicians sometimes have limited statistical training and require more help when preparing bioassay data for filing with regulatory agencies. Three things should be considered when approaching problems in statistics: (i) Assess the situation: data should be checked for accuracy and bias; (ii) Is the situation resolving on its own? Are the skills in place to answer this question? (iii) If no to 2, find expert help. Some sources of variability in bioassays are listed. Some sources of expert help are listed. An example is given of how a statistics problem was solved by a technician unfamiliar with statistics. Some practical tips for design, analysis and regulatory issues are presented. PMID- 12079193 TI - A case study: development of a bioassay for IL-1ra. AB - The IL-1ra bioassay was overhauled in 1993 at Synergen, Inc. The team included bioassay specialists and two pre-clinical statisticians. We began by focusing on the technical aspects, optimizing the number of cells per well, altering the staining procedure, and checking incubation times. We then proceeded to revise the layout of the dose-response curves on the 96-well plates and included an internal control as well as a standard. Finally, we reconfigured the statistical modelling procedure to model each plate as a whole using a 9-parameter logistic curve. When the work was concluded, the precision (%CV) had improved from 44% to 10%, while the retest rate was reduced from 20-30% to less than 5%. PMID- 12079194 TI - Constrained four parameter logistic model. AB - The constrained four parameter logistic model has found wide application in describing dose response relationships across many assay systems. This discussion examines the basic model and its practical application to potency testing in the context of the 96 well plate. A two step procedure is recommended for the analysis: (i) the constrained logistic model to generate potency estimates, (ii) a linear mixed-effects model to account for within-plate and between plate variability for producing the final combined estimate of potency. The method is outlined in a case study. Design issues related to possible location effects on the plate may be ameliorated by use of a Latin square design. PMID- 12079195 TI - An aspect of discrete data analysis: fitting a beta-binomial distribution to the hospitals' data. AB - Statistical analysis for discrete data, particularly for probability models such as the binomial, Poisson and multinomial, is by now very well understood, with a wealth of suitable software. It can happen that the standard generalized linear modelling (glm) software is not completely appropriate, since over-dispersion is present, relative to the standard distributions such as the Poisson or the binomial. Failure to take account of this over-dispersion, for example in fitting a model such as log(p/(1 - p)) = alpha + beta x (where the covariate x is the dose) will mean that our estimates of beta will be less precise than the binomial based formula gives us. Thus for example we will be quoting confidence intervals for beta that are too narrow. One way of coping with this problem is to use a probability model which is more general than the binomial, and one such model is the beta-binomial. This paper discusses beta-binomial modelling (in S-Plus) in relation to the interesting data set given in the 1998 BMJ paper by Spiegelhalter and Marshall on success rates of 52 in vitro fertilisation clinics in the UK. PMID- 12079196 TI - Methods for the analysis and design of limiting dilution assays. AB - Limiting dilution assays are a useful device for extracting quantitative information when the underlying analytical tool can only provide information on presence or absence. Analysis by maximum likelihood is advocated and some alternative methods are discussed. Several interval estimators, including one based directly on the log-likelihood are presented. Methods for determining designs which permit efficient estimation are considered and some of their weaknesses are mentioned. Opportunities for further methodological development are outlined. PMID- 12079197 TI - Analysis of an in vivo assay for inactivated polio vaccine. AB - Approaches to the statistical analysis of data from the rat bioassay for Inactivated Polio Vaccine are discussed and compared, using data from a recent collaborative study The measured response, an antibody titre obtained from a doubling dilution series, did not satisfy the requirements of normality and homogeneity of variance necessary for the standard parallel line model for quantitative data. Laboratory-specific cut-offs can be defined, to reduce the data to "positive" or "negative" responses, allowing valid analysis with the probit method. PMID- 12079198 TI - Drugs, the human genome, and individual-based medicine. AB - The so-called "Genomic Revolution" has made possible the high-resolution sequencing of the DNA making up the human genome. One of the main conclusions of the currently available sequencing data is that individuals differ genetically from one another via sequence variations in their genomes. When affected genes are transcribed and translated, some of these sequence variations result in protein products that may affect the functioning of the proteins. This has led to widespread optimism that information on an individual's pattern of sequence variations will lead to drugs that target that individual's variant proteins and make "individual-based medicine" possible. I this chapter some of the assumptions underlying the proposed production of individual drug treatments are examined. The assumptions are viewed in the light of very recent experimental evidence about the sequence patterns found in humans. Also discussed are ancillary ethical problems in cataloging and using databases containing individuals' sequence data, what human genomic sequences are revealing about the use of animal models in developing drugs, and how evidence is mounting that the human genome is only one element serving to maintain an organism's interaction with its environment. PMID- 12079199 TI - Potassium channels: gene family, therapeutic relevance, high-throughput screening technologies and drug discovery. AB - Existing drugs that modulate ion channels represent a key class of pharmaceutical agents across many therapeutic areas and there is considerable further potential for potassium channel drug discovery. Potassium channels represent the largest and most diverse sub-group of ion channels and they play a central role in regulating the membrane potential of cells. Recent advances in genomics have greatly added to the number of these potential drug targets, but selecting a suitable potassium channel for drug discovery research is a key step. In particular, the potential therapeutic relevance of a potassium channel should be taken into account when selecting a target for screening. Potassium channel drug discovery is being driven by a need to identify lead compounds that can provide tractable starting points for medicinal chemistry. Furthermore, advances in laboratory automation have brought significant opportunities to increase screening throughput for potassium channel assays, but careful assay configuration to model drug-target interactions in a physiological manner is an essential consideration. Several potassium channel screening platforms are described in this review in order to provide some insight into the variety of formats available for screening, together with some of their inherent advantages and limitations. Particular emphasis is placed on the mechanistic basis of drug target interaction and those aspects of structure/function that are of prime importance in potassium channel drug discovery. PMID- 12079200 TI - Dual serotonin and noradrenaline uptake inhibitor class of antidepressants potential for greater efficacy or just hype? AB - Preclinical and clinical studies support the rationale that development of single molecules, which would promote serotonergic and noradrenergic neurotransmission by inhibiting simultaneously the uptake of both monoamines, would potentially result in improved antidepressant drugs. Currently, the dual inhibitors of serotonin and noradrenaline uptake are venlafaxine, milnacipran and duloxetine. Based on the preclinical studies, the three drugs do show properties of inhibiting uptake of both monoamines in vitro and in vivo in the following order of decreasing potency: duloxetine, venlafaxine and milnacipran, and all exhibit low affinity at neuronal receptors of neurotransmitters, suggesting low side effect potential. In double-blind, controlled studies, venlafaxine and milnacipran were repeatedly shown to be as efficacious as tricyclic antidepressant drugs in treating major depressive disorder, while one double blind, placebo-controlled trial showed the antidepressant efficacy of duloxetine. Specifically designed comparative trials of dual uptake inhibitors against the other agents are needed to establish whether the dual uptake inhibitors show improvement in efficacy, rate of responders, antidepressive effects and/or remission. PMID- 12079201 TI - Advances in QSAR studies of HIV-1 reverse transcriptase inhibitors. AB - A review is presented of the recent advances in quantitative structure-activity relationship (QSAR) studies of HIV-1 reverse transcriptase (RT) inhibitors. These inhibitors have been put into two classes: nucleoside RT inhibitors (NRTIs), which are 2',3'-dideoxynucleoside analogues (ddNs), and non-nucleoside RT inhibitors (NNRTIs). For NRTIs (ddNs), which act as competitive inhibitors or alternate substrates of RT and hence interact at the substrate binding site of the enzyme, QSARs have pointed out the major role of the electronic factors governing their activity. For NNRTIs, which bind to a site entirely distinct from the substrate binding site, the activity has been shown to be largely dependent upon the hydrophobic nature of the compounds or substituents. The hydrophobic nature of the active site in the receptor with which the NNRTIs interact provides relatively few possibilities for the molecules to have polar interactions or hydrogen bondings, but QSARs have indicated that NNRTIs do involve some polar interactions and hydrogen bondings with some pockets of the enzyme. QSARs also indicate the significant roles of steric interactions and conformational shape of the molecule. PMID- 12079202 TI - Herbal medicine of Wisconsin Indians. AB - In this chapter we will familiarize the reader with selected aspects of the herbal medicine of Wisconsin Indians. We will concentrate on the Menominee, Potawatomi, Ojibwe and Meskwaki tribes. We will address the following topics: (1) selected methods for research in the existing literature to reveal both the identity and the chemical composition of plants which Indians used for medicinal purposes; (2) some aspects of Indian medicine, such as the mode of delivery of herbal drugs, extensive use of plant mixtures, and other practices; (3) selected literature resources on comparison of uses of medicinal plants among different North American Indian tribes and contemporary white men. PMID- 12079203 TI - The impact of multiple drug resistance (MDR) proteins on chemotherapy and drug discovery. AB - Transportation of molecules across the cell membrane in living organisms is a critical aspect of life. Transportation includes importation of nutrients from the environment and exportation of toxic compounds. When export includes therapeutic compounds, then the practice of clinical medicine may become compromised. Often efflux of therapeutic compounds is mediated by a large superfamily of proteins referred to as multidrug resistance (MDR) proteins. The initial sections of this chapter are focused on MDR proteins and their negative impact on clinical medicine in cancer chemotherapy as well as infectious diseases mediated by bacteria, fungi and parasites. A brief description of major classes of MDR proteins found in microbes is followed by a more exhaustive treatment of ABC transporters in lower eukaryotes and parasites as well as cancerous mammalian cells. Later sections deal with potential and real positive aspects and applications brought about by a growing knowledge of MDR proteins. Examples described include improved antibiotic production, leveraging MDR proteins in drug discovery, new therapeutic options, dual therapy in treatment of cancer and infectious diseases, and finally MDR proteins as targets for new classes of therapeutic compounds. PMID- 12079204 TI - Loss of heterozygosity and microsatellite instability in acquired melanocytic nevi: towards a molecular definition of the dysplastic nevus. AB - Acquired melanocytic nevi may show signs of histological dysplasia, and epidemiological studies have demonstrated that dysplastic melanocytic nevi (DMN) are associated with an elevated melanoma risk. Nevertheless, the concept of DMN as precursors of melanoma has remained a concept, in view of the difficulty of establishing unambiguous cytological and histological criteria for DMN. Recent molecular data suggest that genetic instability is more frequent in DMN than in benign acquired melanocytic nevi. We have analyzed 54 benign melanocytic nevi and 6 DMN for loss of heterozygosity (LOH) at microsatellite markers D9S171, IFNA, D9S270, D9S265. LOH at one or more loci was detected in 17 out of 54 benign nevi and in 4 out of 6 DMN. LOH was demonstrated at 26 out of 103 amplified and informative microsatellites in benign nevi and at 6 out of 11 microsatellites in DMN. In addition, 6 benign nevi and 6 DMN were microdissected in 4-15 regions per lesion and analyzed for LOH and microsatellite instability (MSI) at D9S162 and D14S53. Both LOH and MSI were detected more frequently in dysplastic nevi (LOH frequency 0.61 vs 0.18; MSI frequency 0.27 vs 0.05). These results confirm that genetic instability is more prevalent in DMN than in benign acquired melanocytic nevi. Therefore, DMN might be defined as a monoclonal and genetically unstable, but limited, melanocytic proliferation that distinguishes this entity from the benign nevus and from malignant melanoma. PMID- 12079205 TI - The changing incidence and mortality of melanoma in Australia. AB - For many years Australia has had the highest incidence and mortality rates in the world for melanoma. The incidence rate has been increasing at around 5% per year and the mortality rate, at a rate slightly lower than that. Epidemiology studies have shown clearly that there is both a constitutional and an environmental contribution to melanoma risk, with sunlight being the major risk factor in the environment. The data also clearly show that the thickness of a melanoma at the time it is removed is one of the major determinants of the likelihood of metastasis and thus of the long-term prognosis. Both of these components have been incorporated into major public health programmes aimed at melanoma control in Australia over the last 25 years. Primary prevention programmes have been aimed at reducing the desire for a tan and subsequent overexposure to sunlight. Secondary prevention (early detection) programmes have encouraged people in the community to seek early attention if they notice a new or changing pigmented lesion. Although the age-adjusted incidence and mortality rates for Australia continue to rise, cohort analysis of both incidence and mortality rates reveals that the overall rise is not reflected in all age groups. In the younger cohorts- groups that it has been possible to influence by our public health campaigns in recent decades--both incidence and mortality rates are dropping. PMID- 12079206 TI - Why is epiluminescence microscopy important? AB - The new morphological information provided by epiluminescence microscopy (ELM) requires a fresh approach to the analysis of pigmented lesions. It necessitates a learning process that pertains to the recognition of hitherto unknown morphological features and is based on the discrimination of these features and their combination into two different patterns. ELM has been shown to improve the sensitivity and specificity of the diagnosis of melanoma and other pigmented lesions by 25-30%. Digitized ELM (DELM) provides an unlimited capacity for data storage and retrieval. It is a computerized imaging method, objective and noninvasive; it provides objective evidence of lesional changes on follow-up; documents growth and any changes in the structure and shape of lesions; and thus helps in decisions on whether to excise them or not. It provides for quality control by means of the aforementioned documentation, which may also serve as back-up in the case of medico-legal problems. In addition, the spectrum is widened by the dimension of teledermatology and cybernet computer-assisted diagnosis, which holds great promise for the future. ELM and DELM are thus the most important single development of the past three decades in the early diagnosis of melanoma. PMID- 12079207 TI - Sentinel node biopsy: not only a staging tool? AB - The management of clinically negative regional lymph nodes in early-stage melanoma has been controversial for at least a century. While some surgeons offer elective lymph node dissection (ELND), others recommend treatment of the primary alone and only perform a therapeutic dissection (TLND) for cases of recurrence in the nodal basin. The rationale for ELND is based on the concept that metastases occur via the sequential passage of tumor from the primary site to the regional lymph nodes and then to more distant sites. If this theory is correct then early dissection of the regional lymph nodes will disrupt the metastatic cascade and prevent further spread of disease. On the other hand, advocates of the "wait and watch" approach suggest that metastases to the regional lymph node basin are only a marker of disease progression and that distant disease can occur in the absence of lymph node metastases. Four randomized prospective studies have examined the efficacy of ELND versus TLND. While all four studies have failed to demonstrate a survival advantage of ELND, there is some suggestion that patients with metastases in the regional basin may benefit from ELND. As an alternative approach to this controversy, Morton and associates at the John Wayne Cancer Institute devised the technique of intraoperative lymphatic mapping and sentinel lymphadenectomy (LM/SL). This minimally invasive operative procedure allows the surgeon to identify the first or sentinel lymph (SN) in the regional basin. The technique is predicated on accurate mapping of the cutaneous lymphatics by lymphoscintigraphy and the intraoperative use of a vital blue dye to lead the surgeon to the SN and allow the pathologists to identify metastases in the lymph nodes. Patients with tumor-positive dissections would undergo complete lymph node dissection (CLND), and for those without metastases the complications and costs associated with CLND could be avoided. The success of the procedure depends on the completion of a learning phase and on the cooperation of nuclear medicine physicians, surgeons, and pathologists. While this technique has become almost standard practice in the United States and around the world, we await the results of several important clinical trials to determine whether LM/SL will replace ELND or the wait and watch approach in the management of early-stage melanoma. PMID- 12079208 TI - Role of clothes in sun protection. AB - Ultraviolet (UV) radiation is the carcinogenic factor in sunlight. Damage to skin cells from repeated UV exposure can lead to the development of skin cancer. Apart from avoidance of the sun, the most frequently used form of UV protection has been the application of sunscreens. The use of textiles as a means of sun protection has been underrated in previous educational campaigns, even though suitable clothing offers usually simple and effective broadband protection against the sun. Apart from skin cancer formation, exacerbation of photosensitive disorders and premature skin aging could be prevented by suitable UV-protective clothing. Nevertheless, several studies have recently shown that, contrary to popular opinion, some textiles provide only limited UV protection. It has been found that one-third of commercial summer clothing items provide a UV protection factor (UPF) less than 15. Given the increasing interest in sun protection, recreationally and occupationally, test methods and a rating scheme for clothing were needed that would ensure sufficient UV protection. Various textile parameters have an influence on the UPF of a finished garment. Important parameters are the fabric porosity, type, color, weight and thickness. The application of UV absorbers into the yarns significantly improves the UPF of a garment. Under the conditions of wear and use several factors can alter the UV protective properties of a textile, e.g., stretch, wetness and laundering. The use of UV-blocking cloths can provide excellent protection against the hazards of sunlight; this is especially true for garments manufactured as UV-protective clothing. However, further educational efforts are necessary to change people's sun behavior and raise awareness for the use of adequate sun-protective clothing. PMID- 12079209 TI - The impact of surgery on the course of melanoma. AB - Skin melanoma, unlike other cancers, occurs on the body surface: it can be detected and treated before it reaches the stage where it can metastasise; the impact of surgery is unrivalled, but only while it is at this early stage. In melanomas more than 0.75-1.00 mm thick, an increasing proportion acquire metastatic properties. There is today evidence showing that wide excision does not help, and that the effect of surgery is limited to local control of the disease. According to randomised trials, the territory of early spread--without concomitant distant micrometastases--that can be eradicated by surgery is shrinking. It has been demonstrated that resection margins of 3-4 cm are no better in terms of recurrence and survival than margins of 1 or 2 cm. Most melanomas can now be adequately resected without skin grafting. Regional elective lymph node dissection for high-risk melanoma (1.5 mm thick or more) does not improve survival over that obtained with delayed lymph node dissection performed when clinical metastases appear. By analogy, prophylactic isolated limb perfusion with melphalan reduces the rate of in-transit metastases but does not improve survival. Sentinel node biopsy allows early detection of regional lymph node metastases with minimally invasive surgery. On-going randomised studies will show whether it can have any impact on survival. Considering the experience with elective lymph node dissection, it seems unlikely that selective--as opposed to elective--lymph node dissection, of positive sentinel nodes, will influence survival. The already extensive experience with sentinel node biopsy provides a death risk hierarchy: one N2 node (with clinical metastasis), one N1 node--or sentinel node--with micrometastasis and one N0 node with no histologically detectable micrometastasis but PCR positive give, respectively, 50%, 60% and 70% 5-year survival rates. In other words, the earlier the detection of metastasis, the longer the survival. In terms of growth kinetics, the earlier the detection of metastasis, the longer the time to death, with no evidence that surgery would have an impact. There is just one, as yet unpredictable, subset of pa- tients with lymph node-confined disease in whom surgery might have an impact. It is hoped that, in the future, gene expression profiles of primary melanoma will help to pick out these patients. Multivariate analysis has shown that the sentinel node status is the most powerful prognostic factor in primary melanoma. Sentinel node biopsy is a valuable tool for selecting patients for adjuvant treatments within the frame of clinical trials, in which micrometastatic and clinically involved lymph nodes are entered separately. In-transit metastases can be eradicated in 50% of cases by isolated limb perfusion with melphalan under mild hyperthermia. When in-transit metastases are recurrent, deep seated, or bulky, the combination of tumour necrosis factor (TNF) with melphalan and interferon gamma yields a complete response rate of around 80%. This is the first antiangiogenic treatment of cancer that is effective in clinical practice, but it has no effect on survival. Current better knowledge of melanoma biology indicates that local, limited surgery has an impact on local or regional spread only. PMID- 12079210 TI - Perspectives of pegylated interferon use in dermatological oncology. AB - The potent immunomodulatory, antiproliferative and antiviral properties of interferons (IFNs), together with their availability in large amounts thanks to the recombinant DNA technique, have resulted in their widespread clinical use in a variety of viral and nonviral proliferative disorders. In dermato-oncology, IFNs have been used primarily in melanoma, but also in nonmelanoma skin cancer, such as squamous and basal cell carcinomas, Kaposi sarcomas and lymphomas. Trials with IFNs have been performed in patients with melanoma in an adjuvant setting (stage II and III) and in metastatic disease (stage IV). While the response rates with IFNs as single agents in stage IV disease usually do not exceed 15%, the use of adjuvant IFNs has been claimed to increase disease-free survival (stage II), or even overall survival (stage III), in low- or high-dose regimens, respectively; the latter, however, involved numerous side-effects and were beset with lack of compliance and acceptance, as well as being very costly. Pegylated IFN (PEG-IFN) is a form of recombinant human IFN that has been chemically modified by the covalent attachment of a branched metoxpolyethylene glycol moiety. Pharmacogenetic and pharmacodynamic data obtained in animal and in phase I studies have indicated that PEG-IFN injected once a week has the potential to be superior in efficacy to human IFN injected three times a week. The safety profiles of PEG-IFN and IFN are comparable in healthy volunteers and in chronic hepatitis C (CHC) patients. PEG-IFN is currently being evaluated for the treatment of CHC, renal cell carcinoma, chronic myelogenous leukaemia, and malignant melanoma, the last in both stage IV and stage III disease. PMID- 12079211 TI - Dendritic cell vaccination for the treatment of skin cancer. AB - Reasons for failure of the immune system to fight cancer include tumor immune escape mechanisms, limited availability of tumor-specific antigens, and failure to deliver tumor antigens in the right immunological context. Progress in molecular biology and immunology has provided technologies that can detect an ever-widening choice of new tumor-specific antigens. One of the most important questions remains the delivery of these tumor antigens in an effective way to the immune system of a cancer patient. This task is performed in normal circumstances by dendritic cells (DCs). DCs are sentinels of the immune system located at sites of antigen entry such as skin. They take up antigen and carry it to secondary lymphoid organs. They are highly specialized antigen-presenting cells (APCs) for the activation of specific effector T cells recirculating in secondary lymphoid organs. In recent years an enormous increase in our understanding of DC biology has opened up new ways of applying these cells for immunotherapy of cancer. PMID- 12079212 TI - Gene-based immunotherapy of skin cancers. AB - Skin cancers continue to present a major therapeutic challenge to physicians. Recent advances in molecular genetics and improved understanding of immune responses to tumors have generated an interest in using gene-based immunotherapy for treating these malignancies. Two major forms of gene-based immunotherapy are currently being investigated. One focuses on genetic modification of some target cell populations of the host using immunostimulatory genes such as cytokines, in order to improve tumor immunogenicity and antitumor responses; the other is genetic immunization with the genes coding for melanoma-associated antigens recognized by cytotoxic T cells. This paper reviews these novel strategies and summarizes the most recent data recorded in either experimental studies or clinical trials. PMID- 12079213 TI - Cytokine fusion protein treatment. AB - Mice suffering from melanoma can be cured by treatment with a recombinant antibody-lymphotoxin-alpha fusion protein. In order to characterize the involvement of T cells in this syngeneic tumor model, the T cell receptor repertoire was analyzed both quantitatively and qualitatively. In this connection, quantitative analysis of the T cell receptors displayed only a modest overexpression in some of the variable beta chain families over the course of therapy. Clonotypic mapping, however, revealed a marked increase in the number of clonally expanded T cells among the tumor-infiltrating T cells, suggesting a specific activation. PMID- 12079215 TI - A rational approach to the follow-up of melanoma patients. AB - There are no generally accepted guidelines for the follow-up of cutaneous melanoma (CM), and there is an ongoing debate about the value of follow-up examinations. Some authors doubt whether early detection has any beneficial effect on patient survival and suggest that it may only prolong the patient's period of suffering from the knowledge of having metastasis. A systematic review of the literature on early detection and resection of CM metastasis shows the following picture: (1) In in-transit metastasis and in regional node metastasis, the tumour volume of the metastatic nodules at the time of diagnosis is prognostically significant. Either the number of nodes involved in regional metastasis or the diameter of the largest node showed prognostic impact in different studies. Therefore, early detection seems to affect the cure rate in this stage of disease. (2) In distant metastasis, surgical resection of all recognisable metastases prolongs survival. This is true as long as only one organ system is involved and particularly if complete resection of all metastases can be achieved. Therefore, early detection contributes to prolongation of survival. We performed a follow-up study in 2008 prospectively documented consecutive patients with stage I-III cutaneous melanoma who presented for follow-up examination at the Department of Dermatology of the University of Tubingen from August 1996 to August 1998. Stage-appropriate follow-up examinations were carried out according to the German Society of Dermatology guidelines. A total of 3,800 clinical examinations and 12,398 imaging techniques were documented: 62 second primary melanomas were detected in 46 patients and 233 disease recurrences in 112 patients during this time. Physical examination was responsible for the discovery of 50% of all recurrences, with the patient initially detecting the metastasis on self-examination in 17% of these cases. Technical examinations were responsible for the detection of the remaining 50%. In the primary tumour stages, 21% of all recurrences were discovered by lymph node sonography, the majority being classified as early detection. Among the recurrences, 48% were classified as early detection, and these patients had a significantly more favourable probability of recurrence-free survival than those with recurrences classified as late detection. The results of our study suggest that a follow-up schedule elaborated for cutaneous melanoma is suitable for the early detection of second primary melanomas and of early recurrences in approximately 5% of patients during a 2-year follow-up period. PMID- 12079214 TI - Cytotoxic T-cell induction in metastatic melanoma patients undergoing recombinant vaccinia virus-based immuno-gene therapy. AB - In an ongoing phase I/II study, metastatic melanoma patients were treated with a replication-incompetent recombinant vaccinia virus (rVV) encoding Melan-A(27-35), gp100(280-288), and tyrosinase(1-9) HLA-A*201-restricted epitopes together with B7.1 and B7.2 co-stimulatory molecules. rVV was administered in the context of systemic GM-CSF treatment. Boosts were subsequently administered 2 weeks apart with corresponding synthetic nonapeptides and GM-CSF. Two cycles of treatment were administered 2 weeks apart from each other. Specific immune responses were evaluated by quantitative assessment of cytotoxic T-lymphocyte precursor frequency and tetramer staining. By the time the two cycles had been completed, four out of five patients showed significant (greater than threefold) increases in gp100(280-288)-specific and four out of five, in Melan-A(27-35)-specific tetramer staining of CD8+ cells. Frequencies of CTL precursors specific for gp100(280-288), tyrosinase(1-9) and Melan-A(27-35) were also significantly increased in all five, and in four and four of the five patients, respectively, in some cases within 12 days after the first injection of the recombinant vector. Thus, the innovative vector under investigation is able to raise a concurrent and specific cellular immune response against a panel of molecularly defined antigens, thereby increasing the chance of an immune hit against neoplastic cells displaying heterogeneous antigen expression. PMID- 12079216 TI - Micrographic surgery of basal cell carcinomas of the head. AB - Basal cell carcinoma (BCC) is a locally invasive malignant cutaneous tumour with a rising incidence. This tumour can be treated successfully by a variety of techniques, including local excision, radiation, cryotherapy, curettage, electrodessication and laser obliteration. Micrographic surgery is a specialised type of minimal marginal surgery that offers higher cure rates than do other options in the treatment of contiguous skin cancers in selected settings. The horizontal frozen histological sections of the excised tumour permit complete microscopic examination of the surgical margin. Maximum sparing of tumour-free adjacent tissue is achieved with histological mapping of the tumour boundaries, and subsequent wound reconstruction is optimised. Data on topographical distribution, histopathological subtype, subclinical tumour extension, therapeutic procedures required for complete eradication, and recurrence rates were recorded in 3065 BCC of the head. Micrographic surgery is the treatment of choice for large or invasive primary BCC with uncertain clinical boundaries, especially in difficult anatomical regions, for recurrent or re-recurrent BCC, and for tumours with an aggressive histopathological pattern. PMID- 12079217 TI - Repair of cutaneous defects after skin cancer surgery. AB - The goal of this chapter is to present an overview of the main clinical applications of surgical repair in the various anatomical regions of the head. Special consideration will be given to the areas that require reconstruction and to possible problems and complications. Although the skin lesion often dictates the type of incision made, there are various options. The biggest advantage of flaps is the excellent aesthetic result; this is feasible because the skin is similar in color and texture to that being replaced, and moreover no contracture occurs. On the other hand, a flap requires additional incisions and tissue movement, which increases the risks of postoperative complications. The range of flaps that can be used is discussed in detail. Full-thickness skin grafts are an important and necessary tissue source for reconstructive surgery in the face. They must meet the functional and aesthetic challenge posed by the surgical defect and are usually chosen because of the lack of adjacent tissue. Whenever possible, the skin grafts should resemble the surrounding skin in texture, color and thickness. PMID- 12079218 TI - Radiotherapy of skin tumors. AB - The incidence of cancers of the skin is increasing, as is life expectancy among most of the population. Besides surgery, all skin cancers can be treated with radiotherapy, with excellent results. Unfortunately, both less training and less equipment are available than earlier, which means that dermatologists also have less experience in this field. We would like to propose radiotherapy for medium sized or larger lesions, especially on the face in elderly people. Good indications are keratoacanthomas, extensive actinic keratoses, Bowen's disease including erythroplasia of Queyrat, basal cell and squamous cell carcinomas, but also lentigo maligna and lentigo maligna melanomas. These tumors can be treated in a curative way. Excellent results of palliative X-ray therapy are achieved in Kaposi's sarcoma and in lymphomas, and also in Merkel cell tumors. After 100 years of treatment of skin cancers by radiotherapy, dermatologists should not forget that if appropriate principles are followed and precautions are taken, X ray treatment is still a safe and effective method. PMID- 12079219 TI - Photodynamic therapy and fluorescence diagnosis of skin cancers. AB - In several countries throughout the world the photosensitizer porfimer-sodium has been approved for systemic photodynamic therapy (PDT) for different oncological indications. However, owing to the prolonged photosensitization entailed, the use of this porphyrin derivative is restricted. Currently, the most promising sensitizers in dermatology that can be applied topically are 5-aminolevulinic acid (ALA) or ester derivatives that are precursors of heme biosynthesis. ALA has shown good clinical and excellent cosmetic results in superficial skin cancer and precancerous conditions, e.g. superficial basal cell carcinoma (BCC), or actinic keratoses (AK): phase III studies have demonstrated its efficacy especially in Bowen's disease and AK. ALA-PDT for AK was therefore approved by the FDA in late 1999, and the corresponding registration process is currently in train in Europe. Besides its usefulness in oncological therapy, ALA also has a unique feature that can be exploited for diagnostic purposes: after topical or systemic application protoporphyrin IX is induced rather selectively in epithelial tumors, with a high tumor-to-surrounding tissue ratio, which can be visualized after excitation with light. By using a CCD camera system together with digital imaging, the contrast of the acquired fluorescence images can be significantly enhanced and allows the determination of a threshold, which can be utilized either for a directed biopsy or for preoperative planning when Mohs' surgery is scheduled. At present, the routine employment of such systems is being assessed in prospective studies. PMID- 12079220 TI - Intralesional interferon in basal cell carcinoma: how does it work? AB - Basal cell carcinoma (BCC) is the most common skin cancer among Caucasians, and its incidence is increasing. Intralesional injection of interferon alpha (IFN alpha) has been shown to provide a safe and effective treatment for BCCs. The predominant mechanism for the effect of IFN alpha on BCC has been partially identified. We have shown that in untreated patients, BCC cells constitutively express CD95 ligand (CD95L), but not the receptor. BCC cells make use of the CD95 ligand to escape from a local immune response by averting the attack from activated CD95 receptor-positive CD4+ T cells. The CD95L of BCC cells is functional as CD95+ target cells incubated on BCC cryosections become apoptotic and are lysed. In IFN alpha-treated patients BCC cells express not only CD95L but also CD95 receptor, and regress by committing suicide or fratricide through apoptosis induction via CD95 receptor-CD95L interaction. Peritumoral infiltrating cells, predominantly CD4+ T cells, may support regression of BCC by the secretion of cytokines such as IFN gamma or interleukin-2 which may also be responsible for the up-regulation of CD95 on BCC cells. PMID- 12079221 TI - Epithelial malignancies in organ transplant patients: clinical presentation and new methods of treatment. AB - Transplantation of solid organs has been well established as a mode of therapy for the treatment of various end-stage organ diseases for many years. Up to now, it has benefited more than 1 million patients worldwide. The long-term success of organ transplantation depends particularly on the prevention of allograft rejection. Various regimens have been used to suppress hosts' cellular immune responsiveness to the grafted organs. Nowadays immunosuppressive therapies consist mainly in prednisolone, azathioprine, cyclosporine, anti-T-lymphocyte globulin (ATG), anti-CD 3 antibody (OKT3) and substances of a new generation, such as tacrolimus or mycophenolic acid. However, not only the patient's reactivity to the graft is impaired, but also that to infectious organisms. Chronically altered immune responsiveness is especially associated with a dramatically increased risk of malignancy, most frequently non-Hodgkin's lymphoma and skin cancer. Within the first 5 years of immunosuppression 40% of transplant recipients experience premalignant skin tumors such as actinic keratoses and Bowen's disease, and also such skin cancers as squamous cell carcinomas and basal cell carcinomas. Quite often these have an aggressive biology and an uncommon morphology. Cancer is now responsible for a mortality rate of 5-8% in organ transplant patients. Various risk factors, such as exposure to sun and infections with oncogenic viruses (e.g. HPV) contribute to the already increased risk of dysplasia when lifelong immunosuppression is required. Prophylactic strategies therefore include the development of virus-like particles (VLPs) as anticancer vaccines, which might become a very interesting approach to preventing HPV associated cancer. The prevention of precancerous conditions and mature skin cancers in grafted patients includes protective clothing and adequate protection of UV-exposed skin regions, including lips, from sunlight with appropriate sunscreen. Close dermatological surveillance through a specialized outpatient department should be ensured to detect potentially fatal skin malignancies at an early stage. Early treatment of precancerous lesions includes topical retinoids, such as tretionin, tazarotene or adapalene. A 5% fluorouracil cream is widely used but shows variable effects on manifest actinic keratoses. As cellular immunity seems to play the major part in the prevention and cure of malignant and premalignant cutaneous neoplasias as well as viral infections, a specific enhancement of the local immunity would be desirable. Imiquimod is one of a class of agents known as immune response modifiers. The drug has been shown to have both antiviral and antitumor activity. Application of immune response activators or modifiers such as imiquimod might be premising in the case of transplant recipients. PMID- 12079222 TI - New treatment modalities for basal cell carcinoma. AB - Basal cell carcinoma (BCC) is a subtype of nonmelanoma skin cancer (NMSC), a potentially fatal disease linked to overexposure to the sun during childhood. BCC has been associated with UV-induced mutations of the PTC and p53 tumor suppressor genes, and to polymorphisms in the melanocortin-1 receptor and XPD genes. Mortality rates due to BCC are low, but its increasing incidence and prolonged morbidity means the disease is costly to treat. Early recognition and effective treatment are therefore important, to reduce the incidence of BCC and lighten the economic burden of its management. This paper reviews current treatments for BCC, including excision and curettage, electrodessication, surgery, cryosurgery, radiotherapy, and treatment with 5-fluorouracil and intralesional/perilesional cytokines. It also deals with two new treatment modalities, photodynamic therapy and imiquimod 5% cream, an immune response modifier that effectively resolves BCC lesions. PMID- 12079223 TI - Sun protective clothing: 5 years of experience in Australia. AB - The Australian/New Zealand Standard AS/NZS 4399 "Sun protective clothing- evaluation and classification" was published in 1996. AS/NZS 4399 has been well accepted and most companies wishing to claim UVR protection for their products have complied with the labeling requirements. This standard is not mandatory, unlike two other Australian standards dealing with solar ultraviolet radiation (UVR) protection, namely the Sunscreen Standard (AS2604) and the Sunglass Standard (AS1067). With these standards there is the ability to impose substantial penalties for non-compliance. In Australia the standard-setting process is achieved by consensus and the development of AS/NZS 4399 was a long and involved process which took a number of years. The standard is not perfect; it was appreciated that issues such as garment lifetime and stretch and wet testing needed to be covered and it was planned to address these in a revised standard. In the 5 years since publication considerable work, in both Australia and overseas, has been carried out. Other national standards have been developed and published. This paper presents some of the rationale which the committee worked through prior to 1996. Also covered are many of the experiences and difficulties in the 5 years since the introduction of AS/NZS 4399, in particular the effect of local conditions and legal requirements on the operation of the standard. PMID- 12079224 TI - From inflammation to neoplasia: new concepts in the pathogenesis of cutaneous lymphomas. AB - Mycosis fungoides is a clinicopathologic term which describes a neoplasm of cerebriform T lymphocytes that form plaques and tumors. We further suggest that mycosis fungoides arises in a background of chronic inflammation or as a response to chronic antigenic stimulation. Subsequently, a series of mutations results in the stepwise progression from eczematous patches, to plaques, tumors and eventual hematogenous dissemination. The pathogenetic process is driven by various, probably individually different, exogenous factors, e.g. environmental foreign antigens, bacterial superantigen, and/or endogenous factors, e.g. autocrine cytokine loops, CD40/CD40L and B7/CD28 interaction. PMID- 12079225 TI - Cutaneous lymphomas and pseudolymphomas: newly described entities. AB - This chapter summarizes some recently described cutaneous lymphomas and pseudolymphomas with regard to their clinicopathological presentation, biological behavior and classification place. Among cutaneous lymphomas, the group of cytotoxic lymphomas, angioimmunoblastic T-cell lymphoma, intravascular lymphoma, hydroa-like lymphoma, marginal zone lymphoma, spindle-cell B-cell lymphoma, and B cell lymphoma with a dermatomal distribution are presented. In the context of pseudolymphomas, cutaneous follicular lymphoid hyperplasia with monotypic plasma cells, pleomorphic reactions in molluscum contagiosum, and CD30+ reactions to parapoxvirus are discussed. PMID- 12079226 TI - Clinical aspects and pathology of primary cutaneous B-cell lymphomas. AB - Primary cutaneous B-cell lymphomas (pCBCL) occur more frequently than generally believed. The most important subtypes are: marginal zone B-cell lymphoma/immunocytoma, follicle center-cell lymphoma and large B-cell lymphoma of the leg. A correct diagnosis can be rendered only in the context of knowledge of the clinical findings. Progress in terms of classification and biology is associated with the application of modern techniques including immunohistology and laser beam microdissection followed by molecular analysis. Future definitions of pCBCL will be based on their molecular abnormalities and their etiology and pathogenesis. Awareness of the special clinical behavior of pCBCL should prevent unnecessarily aggressive treatment. PMID- 12079227 TI - Public education in skin cancer world-wide. AB - Primary and secondary prevention of skin cancer through public and professional education is a major task for the future. During a satellite symposium, skin cancer prevention campaigns in various countries and continents were presented. PMID- 12079228 TI - Modern diagnosis of cutaneous lymphoma. AB - The issue of primary cutaneous lymphoma (CL) has been greatly influenced by the increasing knowledge about lymphoid cell biology, the widespread use of sensitive and specific immunological and molecular markers, and the careful correlation among clinical, histological and immunomolecular features. This latter is the key element of the classification of CL proposed by the EORTC Cutaneous Lymphoma Study Group in 1997, which categorizes distinct clinicopathological entities with prognostic and therapeutic relevance. With few exceptions, no reliable diagnosis and subtyping of CL can be made without the aid of immunohistochemistry and/or molecular analysis. On the other hand, the acritical use of immunological and molecular markers can be misleading if not combined properly with a correct clinical and histological evaluation. For this reason, a step-by-step diagnosis and staging protocol is exceedingly important. Finally, great caution should be used in the interpretation of lymphoid cell infiltrates of the skin which show a monoclonal rearrangement in the absence of reliable clinical and/or pathological evidence of neoplasia. PMID- 12079229 TI - Treatment of cutaneous T cell lymphoma: 2001. AB - The neoplastic cells of mycosis fungoides (MF) and Sezary syndrome are recognized to be clonal expansions of "memory" T cells that home into the upper dermis and epidermis via the interplay of adhesion molecules and chemokines, and this may account for the marked effectiveness and even curative potential of various skin directed therapies utilized to treat clinically early disease. However, because neoplastic T cells freely circulate and are detectable in extracutaneous tissues by PCR prior to other methods, effective long-term control of more advanced MF and Sezary syndrome, i.e. reduction of tumor burden and decreased risk of transformation into aggressive lymphoma, often requires long-term administration of a therapy with systemic activity in addition to skin-directed therapies. Therapies with immunomodulatory activities, e.g. interferon alfa, bexarotene or extracorporeal photochemotherapy (ECP), are probably superior to traditional cytotoxic drugs in this regard although the response rates are relatively low. Selected patients with advanced or biologically aggressive CTCL should be considered for an allogeneic stem cell transplantation regimen to induce a graft versus-tumor response. New and emerging treatments include immunotoxins such as denileukin diftitox that selectively target neoplastic T cells, potent immunomodulatory cytokines such as rIL-12 to enhance innate antitumor immune mechanisms, novel immunotherapeutic approaches that use dendritic cells loaded with tumor-associated antigens or vaccination using synthetic peptides or DNA plasmids that express the variable region of the T cell receptor beta chain, and possibly gene and protein transduction therapy to correct intracellular defects in neoplastic T cells. Overall the future of therapy for CTCL seems quite optimistic. PMID- 12079230 TI - New biologic agents for the treatment of cutaneous T-cell lymphoma. AB - Cutaneous T-cell lymphoma (CTCL) is typically a skin-infiltrating malignancy of clonally derived CD4+ T lymphocytes. Because the host antitumor response appears to play an important role in disease control, systemic therapeutic agents are used in such a manner as to preserve the integrity of the host antitumor response while selectively targeting the malignant cells. The new biologic response modifying treatment options currently used to treat CTCL are reviewed. PMID- 12079231 TI - Etiology and pathogenesis of Kaposi's sarcoma. AB - OBJECTIVE: To provide an overview of the role for HHV-8 as the causative agent responsible for Kaposi's sarcoma, the molecular cross-talk between HHV-8 and HIV 1, and the ability of specific cytokines to influence the pathophysiology of this malignancy. PATIENTS: Normal human skin grafts were studied using a SCID mouse xenograft model system in which engrafted skin was injected intradermally with HHV-8 and/or HIV-1, and the subsequent induction of clinical, histologic, and viral titer changes were assessed in serial fashion. RESULTS: Following intradermal injection of HHV-8 into engrafted normal human skin, cutaneous lesions resembling Kaposi's sarcoma were created in numerous different grafts. These lesions were characterized by routine light microscopy as well as by immunophenotypic and molecular virological assessments. As several grafts injected with HHV-8 failed to develop Kaposi's sarcoma, we sought to determine whether another cofactor was required and this led us to uncover the ability of HHV-8 and HIV-1 to reciprocally influence each other using both in vitro and in vivo studies. Given the importance of various cytokines, the influence of scatter factor and IFN-gamma in the pathophysiology of Kaposi's sarcoma was also evaluated. CONCLUSION: Based on these in vivo studies using SCID mice engrafted with human skin and injected with HHV-8, we conclude that HHV-8 is the etiologic agent in Kaposi's sarcoma, and that HIV-1 and HHV-8 can influence each other involving both direct and indirect cross-talk mechanisms. PMID- 12079232 TI - Connective tissue tumors. AB - Connective tissue consists of collagen, elastic fibers and ground substances produced by fibrocytes. These cells are usually spindle-shaped with slender nuclei and bipolar cytoplasmic extensions. Apart from labeling for vimentin and variable reactivity for factor XIIIa and CD34, fibrocytes are immunonegative. Electron microscopy reveals prominent endoplasmic reticulum, but is otherwise indistinct. Lesions with fibrocytic differentiation can be divided into five categories: scars, keloids, dermatofibromas, nodular fasciitis, and superficial fibromatoses are inflammatory lesions. Thereby, dermatofibromas and their subcutaneous/deep soft tissue counterpart nodular fasciitis can present with a wide variety of clinicopathologic variants which may be misinterpreted as malignancies. Prurigo nodularis, chondrodermatitis nodularis helicis, acanthoma fissuratum, and knuckle pads are hyperplasias; fibroma molle, fibrous papules, connective tissue nevi, and elastofibroma are hamartomas; and fibroma of tendon sheath, pleomorphic fibroma, and giant cell tumor of tendon sheath are benign neoplasms. Deep fibromatoses, dermatofibrosarcoma protuberans, giant cell fibroblastoma, giant cell angiofibroma, hyalinizing spindle cell tumor with giant rosettes, solitary fibrous tumor, myxofibrosarcoma, low-grade fibromyxoid sarcoma, acral myxoinflammatory fibroblastic sarcoma, and classical fibrosarcoma, are malignant neoplasms, that is fibrosarcomas of variable malignant potential. Lesions dominated by myocytes/ myofibroblasts, e.g. cutaneous myofibroma/infantile myofibromatosis, or by macrophages, e.g. xanthogranulomas, are not part of this chapter. PMID- 12079233 TI - European standards for protective apparel against UV radiation. AB - The first European standard which describes the test procedure to determine the UV-protection factor of clothing is about to be completed. A second part of the same standard, dealing with labelling and marking aspects, is ready to be submitted to public enquiry. In this effort a group of experts from most EU member states have cooperated with a high degree of consensus. In this chapter we explain this European standard together with the standard developed in the UK. PMID- 12079234 TI - How to identify patients in need of psychological intervention. AB - The identification of skin cancer patients in need of psychosocial intervention is a necessary prerequisite to relieving their distress and specifically supporting their coping strategies. The influence of early support even on prognosis has been shown in melanoma patients. Only a few patients ask for support themselves. The workload of doctors and nurses limits their ability to identify patients in need of support. Conducting interviews during daily hospital routine is time-consuming and cannot be implemented adequately. The screening of psychosocial risk patients needs to be based on standardized instruments. They should be easy to implement during the every-day routine. The Hornheide questionnaire with 27 items, the short form of the questionnaire with 9 items, and the Hornheide Screening Instrument (HSI) with 7 interview questions have been developed to achieve this aim in skin cancer patients. The Hornheide questionnaire offers a differentiated analysis of distress, and facilitates an accurate intervention according to the individual distress of the patient. The short form does not provide a differentiated analysis of distress. Its advantage is the speedy identification of patients at psychosocial risk. The HSI is for use during the anamnestic interview. In correlation with reduction of hospitalization time the HSI is most practicable for routine clinical implementation. PMID- 12079235 TI - Psychotherapeutic interventions in melanoma patients. AB - OBJECTIVES: The need for and the effectiveness of psychotherapy in the treatment of patients with skin cancer have been shown. However, only insufficient data are available to answer the questions as to what kind of psychotherapy is useful and how psychotherapeutic interventions should be designed. METHODS: During the past 7 years we performed crisis intervention, focal therapy and longer-lasting psychodynamically orientated psychotherapy within a consultation liaison service with the Department of Dermatology of Innsbruck University Hospital. We investigated the type of interventions using qualitative content analysis of the written records of weekly supervisory sessions. RESULTS: Standard methods of psychotherapy are often not appropriate for the confrontation of tumor-related fears in melanoma patients. Modifications of psychotherapeutic interventions and more structured support are necessary to meet overwhelming negative emotions caused by the existential threat of the disease. Therefore, we turned to using expressive-supportive methods combining psychodynamic psychotherapy with relaxation, imaginative methods, and structured picture drawing. CONCLUSIONS: Psychotherapists treating skin cancer patients should receive special education in such methods. To better cope with distressing emotions and to avoid burn-out and withdrawal from severely ill or dying patients, psychotherapists should take part in peer supervision regularly. PMID- 12079236 TI - American standards for UV-protective textiles. AB - During the last 3 years, three standard documents that pertain to the testing and labeling of UV-protective textile products have been published by the American Society for Testing and Materials (ASTM) and the American Association of Textile Chemists and Colorists (AATCC). The titles of these documents, which are available for purchase at www.astm.org and www.aatcc.org are: ASTM D 6544 "Standard Practice for the Preparation of Textiles Prior to UV Transmission Testing", AATCC 183 "Test Method for Transmittance or Blocking of Erythemally Weighted Ultraviolet Radiation Through Fabrics", and ASTM 6603 "Standard Guide to Labeling of UV-protective Textiles". This chapter summarizes the content of each document and shows how the documents are linked together to make a comprehensive plan for the testing and labeling of UV-protective textile products to be sold in the United States. It also describes the intended future work in the United States on UV-protective textile standards. PMID- 12079237 TI - Activities of CIE DIV-6 (photobiology and photochemistry) in UV protection and clothing. AB - Clothing can provide substantial protection against solar ultraviolet radiation (UVR). A technical committee (TC6-29), formed by experts in the field of UVR and photoprotection, was raised and, after extensive exchanges of information on the various existing test methods, prepared a technical report. The report is circulating within the CIE national committees for approval which is expected before the end of 2001. P. Gies (Australia) was in charge of collecting all information and prepared the final document. In the report, various test methods for measurement of UVR transmittance through fabrics are discussed. The measured transmittances can be used to calculate the erythemally weighted UVR transmitted by the fabric and thus the amount of protection provided. Factors affecting the UVR transmission of fabric, i.e. the characteristics of the radiometer, weave, color, weight, stretch, water, quality (holes) and eventual UV-absorbers are also detailed. In vivo and in vitro tests were found to be in broad agreement, particularly when the test method detailed in the AS/NZS 2604 "Sunscreen products -evaluation and classification", with the fabric substituted for the sunscreen, was used. The report concludes: "The UVR transmission of fabrics depends on too many factors to be predicted and must be measured. Particular attention must be paid to sampling to account for variations due to weave and non-uniformity of the product. Accurate and reliable assessment of the protection factors requires spectral measurements of the total (ie. direct and scattered) UVR transmission. A detection system, which closely matches human skin response, can be used to determine protection factors, but should always be checked against the spectral transmission measurement." PMID- 12079238 TI - Comparison of methods: determination of UV protection of clothing. AB - Based on spectrophotometric measurements and mathematical calculations, the ultraviolet (UV) protection factor of a textile is determined in vitro. This technique is the most established test method for the determination of UV protection of a garment. However, the validity and practicality of the in vitro UV protection factor (UPF) determined in the laboratory has been a controversial issue with regard to its significance in the field. Several studies have verified the in vitro UPF by comparing it with various in vivo test protocols using solar simulated radiation for the determination of the minimal erythema dose. The data inconsistency between these studies is certainly due to different methodology. Furthermore, UV dosimetry is a suitable method for quantifying UV transmission through a garment. Chemical dosimeters (e.g. polysulfone films) and biological UV detector films have been used in in vivo-simulated studies in the form of small portable badges monitoring solar UV transmittance through garments on manikins and mobile subjects. As sunlight consists to a considerable extent of diffuse radiation, which is more scattered and absorbed by the fabric than direct radiation, UPF values obtained by measurements in real exposure situations are usually higher than those obtained by conventional in vitro and in vivo testing with collimated radiation beams. Thus the discrepancy between laboratory-based testing and field-based measurements may be due to different radiation geometry of UV sources. Taken together, the in vitro method is the most practicable and inexpensive method for routine measurements of UPF, but dosimetry seems to be a highly useful method for determining the UPF in real exposure situations. PMID- 12079239 TI - Improving UV protection by clothing--recent developments. AB - The assessment of UV transmittance of clothing and the determination of the UV protection factor (UPF) are now well established and the influencing factors such as type of fiber, color, and fabric construction are known. Quick and reliable instruments to measure UV transmittance are crucial. Besides expensive scientific laboratory instruments, a low-cost UV meter is now available for this purpose. The questions arise as to what can be done about a given garment and whether there are ways to improve textiles by the consumer. The many opportunities to improve UV protection of clothing along the textile chain of manufacturing are discussed. The latest possibility for improving the UV-protective properties of clothing is now available at the fabric care stage in every household. A UV absorber can be brought into contact with a fabric during the wash or rinse cycle of a laundry operation. The high UV transmittance of 30% of a thin, bleached cotton swatch in the dry state (UPF 3), can be reduced tenfold to about 3% (UPF >30) in ten washes cycles. This is more than the effect achieved by dyestuffs. The detergent should contain about 0.1-0.3% of the special UV absorber. The same effect can be achieved as early as after one wash cycle with a higher concentration provided by a special laundry additive. Yet another form of application is via rinse cycle fabric conditioner. To make these new types of improvement of fabrics visible the Skin Cancer Foundation now provides the possibility for laundry products to qualify for the "Seal of Recommendation". PMID- 12079240 TI - UV protection and skin cancer. AB - In discussions amongst the public and the scientific community, doubts are repeatedly raised concerning the efficacy of sunscreens in preventing cutaneous malignancy. This article summarizes the most reliable references on UV protection and epithelial skin cancer and discusses the role of UV protection in melanoma prevention. We conclude that there is substantial evidence that UV protection is able to reduce the risk of actinic keratosis, squamous cell carcinoma and probably also the risk of melanoma. PMID- 12079241 TI - Dress up for sun protection/creation of public awareness. AB - A clear or white skin was a sign of distinction for millennia. However, white people, during the last century, sought a tan, which was thought a sign of good health and upper social class. Sun-seekers are now better aware of the risks to their health of UV exposure, but their behaviour has not improved significantly at a global level. The only country deviating from this observation is Australia where aggressive prevention campaigns, such as the "Slip, Slop, Slap" and "Sun Smart" campaigns, have been conducted with success over more than 30 years. However, the financial cost of such campaigns is high, and may be considered too high in countries where skin cancers are fortunately less frequent and/or the culture of public health is unfortunately less developed. The global solar UV index (UVI) program is a response to the increasing skin cancer rate in all white populations. Lead by WHO, the UVI program aims to help people to evaluate the maximum intensity of UV radiation on a given day and at a given location. Since the beginning of the 1990s, UVI forecasts have increasingly been broadcast with weather bulletins. In the matter of protection of the skin, the entire scientific community has agreed that avoiding sun exposure during the 3 to 5 h around noon and to be fully clothed should be recommended. However, for a lot of people to protect the skin means only to apply sunscreen, and these individuals need convincing that it is preferable to use clothes. It may be necessary to involve public health actors in the increasing efforts to educate people and persuade them to adapt their protection strategy to their skin type, to the UV intensity and to their daily activities. PMID- 12079242 TI - The precursors of malignant melanoma. AB - The precursors to melanoma are generally considered to be related to nevi of different types. Here we emphasize the dysplastic nevus, the congenital nevus, and lentigo maligna as specific lesions. The dysplastic nevus is discussed not only as a formal precursor but also as a marker of cutaneous melanoma. The clinical and histologic characteristics are outlined, as well as evidence of progression in dysplastic nevi. The congenital nevus is briefly reviewed and emphasis is placed upon clues to malignant degeneration. The concept of lentigo maligna as a precursor as distinct from an in situ phase is detailed. PMID- 12079243 TI - Interferon-a sensitivity in melanoma cells: detection of potential response marker genes. AB - Interferon alpha (IFN-alpha) represents an adjuvant therapy of proven effectiveness in increasing disease-free interval and survival in subgroups of melanoma patients. Since high doses of cytokine are required, the treatment is often accompanied by toxic side effects. In addition, naturally occurring insensitivity to IFN-alpha may hamper its therapeutic efficacy. Clinical, molecular or immunological markers enabling the selection of potential responders have not so far been identified. To explore the molecular basis of IFN-alpha responsiveness, we analyzed the expression pattern of about 7000 genes in IFN alpha-sensitive and IFN-alpha-resistant cell lines using high-density oligonucleotide arrays. Melanoma cell lines were screened for their sensitivity to proliferation inhibition and HLA class I induction by IFN-alpha by standard 3H thymidine incorporation and flow cytometry. Total cellular RNA from four sensitive and two resistant cell lines was extracted, reverse-transcribed and hybridized to high-density oligonucleotide arrays. The comparative analysis of gene expression in either set of cell lines allowed the identification of four genes (RCCl, IFI16, hox2 and h19) preferentially transcribed in sensitive cells and two (SHB and PKC-zeta) preferentially expressed in resistant cells. These data may provide a useful basis for the development of diagnostic tools to select potential IFN-alpha responders as eligible for treatment, while avoiding unnecessary toxicity to nonresponders. PMID- 12079244 TI - Molecular cytogenetics as a diagnostic tool for typing melanocytic tumors. AB - The melanocyte can give rise to a variety of both benign and malignant lesions that differ in their clinical and histopathological appearance. It is likely that genetic changes underlie this phenotypic diversity. Comparative genomic hybridization (CGH) is a genome-wide scanning technique that permits the measurement of copy number aberrations in archival tumors. Using CGH, we have demonstrated significant differences in the frequency of chromosomal aberrations in primary cutaneous melanomas and Spitz nevi. Whereas the majority of melanomas have aberrations frequently involving chromosomes 9, 10, 7, and 6, most Spitz nevi do not show aberrations. However, a small subset of Spitz nevi show an isolated gain of the short arm of chromosome 11p. As this aberration has not been observed in melanomas, the measurement of chromosomal aberrations should be further evaluated as a diagnostic tool for ambiguous melanocytic tumors. PMID- 12079245 TI - West Nile Virus activity--United States, 2001. AB - In 2001, West Nile virus (WNV) activity was reported from 359 counties in 27 states and the District of Columbia (DC) to ArboNET, a web-based, surveillance data network maintained by 54 state and local public health agencies and CDC. This activity represented a marked increase from 2000, when WNV activity was reported from 138 counties in 12 states and DC. This report summarizes surveillance data for 2001, which indicate that 66 human illnesses were reported from 10 states and that widespread WNV activity in birds, horses, and mosquitoes extended into the midwestern United States and several southern states unaffected previously. The findings in this report underscore the need for public education, increased WNV surveillance aimed at early viral detection, and sustained, integrated mosquito-control activities. PMID- 12079246 TI - Disseminated infection with Simiae-Avium group mycobacteria in persons with AIDS- Thailand and Malawi, 1997. AB - Persons with advanced human immunodeficiency virus (HIV)-1 infection are susceptible to disseminated mycobacterial infections. In the United States, most such infections are caused by Mycobacterium avium or M. intracellulare (i.e., M. avium complex [MAC]). In less developed countries, M. tuberculosis is equally or more prevalent than MAC in persons with HIV-1 infection. Other mycobacterial species have been reported to cause disseminated infection in HIV-infected persons, including Simiae-Avium (SAV) group mycobacteria. SAV group organisms share characteristics of M. avium and M. simiae. Although disseminated (i.e., the isolation of a mycobacterial species from the blood) infection with M. simiae has been reported in HIV-infected persons, another distinct species within the SAV group, M. triplex, was characterized in 1996. Two cases of disseminated infection caused by M. triplex have been reported in HIV-1-positive persons. This report describes four HIV-infected patients from Bangkok, Thailand, and Lilongwe, Malawi, who were infected with SAV group organisms. Because different mycobacterial species are not susceptible uniformly to antimycobacterial agents, accurate identification of mycobacterial species causing an infection is crucial for directing appropriate therapy. PMID- 12079247 TI - Update: Influenza activity--United States and worldwide, 2001-02 season, and composition of the 2002-03 influenza vaccine. AB - The 2001-02 influenza season in the United States was mild to moderate. Influenza A (H3N2) viruses predominated, but influenza B viruses were identified more frequently than influenza A viruses toward the end of the season. Worldwide, influenza A (H3N2) and B viruses predominated.This report summarizes influenza activity in the United States (September 30, 2001-May 18, 2002) and worldwide (October-May) during the 2001-02 influenza season and describes the composition of the 2002-03 influenza vaccine. PMID- 12079248 TI - Predictors of motivation to change after medical treatment for drinking-related events in adolescents. AB - Characteristics associated with intention to change drinking were examined in 254 adolescents treated in an emergency department for alcohol. Younger age, living at home, lower baseline level of drinking and drinking problems, greater depression, having penalties for breaking family drinking rules, higher injury severity, being frightened, and being admitted to the hospital predicted greater intention to change drinking at the time of the event. Baseline characteristics related to being in the action stage of change 3 months later were younger age; lower drinking; having penalties for breaking family drinking rules; injury severity; and number of anticipated consequences, including being in trouble with parents. This study has implications for brief interventions that capitalize on potential teachable moments with problem-drinking adolescents. PMID- 12079249 TI - Self-control and alcohol restraint: an initial application of the self-control strength model. AB - Individuals whose self-control strength is depleted through the prior exertion of self-control may consume more alcohol in situations that demand restraint. Male social drinkers either exerted self-control by suppressing their thoughts or did not exert self-control while doing arithmetic. They then sampled beer. Participants expected a driving test after drinking and therefore were motivated to limit their intake. Individuals who suppressed their thoughts consumed more and achieved a higher blood alcohol content than those who did arithmetic. The groups did not differ in mood, arousal, or frustration. Individuals higher in trait temptation to drink consumed more after suppressing their thoughts relative to those lower in trait temptation. Alcohol intake may be a function of temptation to drink and self-control strength. PMID- 12079250 TI - Self-administration of alcohol before and after a public speaking challenge by individuals with social phobia. AB - K. Abrams, M. Kushner, K. Medina, and A. Voight (2001) showed that alcohol attenuates social anxiety symptoms in socially phobic individuals. This article examines whether social anxiety symptoms can lead to increased alcohol use in this same population. Forty-four individuals with social phobia attended 2 laboratory sessions, spaced 1 week apart, in groups of approximately 10. Participants underwent a social anxiety challenge during 1 session and a control task during the other. Half of the sample self-administered alcohol immediately before, and half immediately after, these 2 activities. As predicted, participants consumed more alcohol following the anxiety challenge than following the control task; however, the opposite pattern was evidenced for drinking preceding [corrected] the 2 activities. These findings add to an understanding of why social phobia and alcohol problems tend to co-occur. PMID- 12079251 TI - Longitudinal substance initiation outcomes for a universal preventive intervention combining family and school programs. AB - This study evaluated the substance initiation effects of an intervention combining family and school-based competency-training intervention components. Thirty-six rural schools were randomly assigned to 1 of 3 conditions: (a) the classroom-based Life Skills Training (LST) and the Strengthening Families Program: For Parents and Children 10-14, (b) LST only, or (c) a control condition. Outcomes were examined 1 year after the intervention posttest, using a substance initiation index (SII) measuring lifetime use of alcohol, cigarettes, and marijuana and by rates of each individual substance. Planned intervention control contrasts showed significant effects for both the combined and LST-only interventions on the SII and on marijuana initiation. Relative reduction rates for alcohol initiation were 30.0% for the combined intervention and 4.1% for LST only. PMID- 12079252 TI - Effect of depressive symptoms on smoking abstinence and treatment adherence among smokers with a history of alcohol dependence. AB - This study examined the effect of depressive symptoms on smoking abstinence and treatment adherence among smokers with a past history of alcohol dependence. Participants (24 women, 27 men) were randomly assigned to behavioral counseling (BC) or behavioral counseling plus cognitive-behavioral mood management training (CBT). The Hamilton Rating Scale for Depression (HRSD; M. Hamilton, 1967) was administered to assess baseline depressive symptoms. Participants who received CBT and had higher HRSD scores were more likely to achieve short-term abstinence from smoking and attend more treatment sessions than those with lower depression scores, whereas for BC participants the effect of HRSD scores was the opposite. Smokers with a history of alcohol dependence reporting high levels of depressive symptoms may benefit from a mood management intervention. PMID- 12079253 TI - Development and validation of the Gamblers' Beliefs Questionnaire. AB - The Gamblers' Beliefs Questionnaire (GBQ) is a self-report measure of gamblers' cognitive distortions. GBQ test items were constructed on the basis of theory, empirical evidence, and expert review. Four hundred three adults completed the initial set of items, and 21 items were selected to make up the final GBQ. The factor structure of the GBQ consisted of 2 closely related factors: Luck/Perseverance and Illusion of Control. The full scale showed good internal consistency (alpha = .92) and adequate test-retest reliability (r = .77). Problem and pathological gamblers scored higher than nonproblem gamblers on the GBQ and its factors. GBQ scores were moderately correlated with the duration of gambling sessions among problem and pathological gamblers, and there was no relationship between GBQ scores and social desirability. PMID- 12079254 TI - The effect of anticipatory strategies on the first day of smoking cessation. AB - The authors tested a model hypothesizing the predictors and the effects of anticipatory strategy use on the 1st day of smoking cessation using data from 63 participants in an ecological momentary assessment study of smoking cessation. Remaining abstinent on the 1st day of cessation was not associated with mean level of urges to smoke during tempting situations but was associated with anticipatory strategy use, which accounted for 18% of the variance. Getting rid of cigarettes contributed the most to this effect (beta = -.397). Getting rid of cigarettes was itself predicted by 2 variables, lower nicotine dependence (beta = -.264) and attending a formal smoking cessation class (beta = .305), which accounted for 15% of its variance. PMID- 12079255 TI - Does alcohol use mediate the association between alcohol evaluations and alcohol related problems in adolescents? AB - Alcohol use becomes more frequent during adolescence, with the percentage of youth who drink doubling from 8th grade to 12th grade. The escalation in drinking behavior during adolescence is often associated with increased problems and other risk behavior, such as drunk driving. In this study, adolescents (N = 557) were recruited from an emergency department to assess their alcohol use, positive and negative evaluations about alcohol, driving after drinking, and problems experienced from drinking. Analyses explored the mediational role of drinking behavior between adolescents' evaluations and problems from drinking and between evaluations and driving after drinking. The findings indicated that drinking behavior partially mediated the association between positive evaluations and problems experienced from drinking but did not mediate the association between evaluations and drunk driving. PMID- 12079257 TI - The inventory of drug use consequences (InDUC): test-retest stability and sensitivity to detect change. AB - The Inventory of Drug Use Consequences (InDUC) is a self-administered measure of consequences of drug use. Psychometric characteristics have been published for its alcohol-focused parent instrument (Drinker Inventory of Consequences; W. Miller, J. Tonigan, & R. Longabough, 1995). Two studies provided information about the test-retest reliability (lifetime) of InDUC scales (Study 1, N = 34) and the replicability of findings to the 5 InDUC scales (recent; Study 2, N = 208). Four scales had good to excellent test-retest reliability. Findings also indicate that reductions in drug use and problems after treatment are large and that the magnitude of change differs between drug use and consequences. Consequences of drug use should be measured directly rather than be inferred from measures of use. PMID- 12079256 TI - Attention-deficit/hyperactivity disorder and conduct disorder symptomatology in adolescents with alcohol use disorder. AB - This study tested the hypothesis that attention-deficit/hyperactivity disorder (ADHD) and conduct disorder (CD) comorbidity is associated with substance use and deviance severity in 395 adolescents with alcohol use disorder. Thirty percent of the adolescents had high ADHD symptom counts, and 73% had 3 or more CD symptoms. ADHD-CD was associated with nonalcohol substance use disorder, drinking levels, and CD severity, but in general substance use was not uniquely elevated or problematic among the comorbid cases. In general, CD and CD severity were more important. The findings did not differ between boys and girls, revealing that in a treatment sample of adolescents, ADHD-CD comorbidity may need to be assessed and treated, but it is not broadly indicative of severity. PMID- 12079258 TI - Relationship among sex, imagery, and exercise dependence symptoms. AB - There is limited research examining the physical (e.g., sex) and psychological correlates (e.g., imagery) of exercise dependence despite its harmful effects. The purposes of this study were to examine sex differences and the predictive ability of exercise imagery for exercise dependence symptoms. Participants were 408 university students who completed measures of exercise imagery, exercise behavior, and exercise dependence. The results indicated that men reported more exercise dependence symptoms than women. For the women, exercise behavior, appearance imagery, and energy imagery were positive predictors of exercise dependence symptoms. In contrast, for the men, exercise behavior and energy imagery positively predicted exercise dependence symptoms. Consistent with suggestions by C. Hall (1995), exercise imagery may be related to exercise dependence symptoms. PMID- 12079259 TI - Adolescents' reactions to the imagery displayed in smoking and antismoking advertisements. AB - This study compared adolescents' unbiased perceptions of the images displayed in smoking and antismoking advertising. Twenty-nine adolescents (ages 11-17) were shown images taken from both advertising types; all images were digitally edited so that no product information appeared in them. Participants described each image in a free-response format and rated each image on self-report dimensions. Content analyses of free-response descriptions and analyses of self-reports revealed that adolescents viewed images taken from cigarette advertisements more positively compared with images taken from antismoking advertisements. These findings suggest that I reason for the potency of cigarette advertising, compared with antismoking advertising, is the inherent positive appeal of the images displayed. Antismoking advertising may be more effective at limiting adolescent smoking if the images displayed have a more positive valence. PMID- 12079260 TI - Negative affect combines with smoking outcome expectancies to predict smoking behavior over time. AB - The present study examined whether the tendency to experience negative affective states combines with smoking outcome expectancies to predict smoking behavior over time. Participants were 121 young adults and resource people recruited from 3 alcohol and drug treatment programs and through community advertisements. Each participant completed 3 interviews over a 4-year period. Results indicated that dispositional negative affect and positive smoking expectancies were significantly correlated with smoking behavior both within and across time. Expectations of positive and negative reinforcement partially mediated negative affect's relation with smoking across time. Positive expectancies did not function as a moderator of negative affect's relation with smoking behavior. These results represent an important step in incorporating smoking outcome expectancies into multivariate models of smoking risk. PMID- 12079261 TI - The human startle reflex and alcohol cue reactivity: effects of early versus late abstinence. AB - This study investigated the human eyeblink startle reflex as a measure of alcohol cue reactivity. Alcohol-dependent participants early (n = 36) and late (n = 34) in abstinence received presentations of alcohol and water cues. Consistent with previous research, greater salivation and higher ratings of urge to drink occurred in response to the alcohol cues. Differential salivary and urge responding to alcohol versus water cues did not vary as a function of abstinence duration. Of special interest was the finding that startle response magnitudes were relatively elevated to alcohol cues, but only in individuals early in abstinence. Affective ratings of alcohol cues suggested that alcohol cues were perceived as aversive. Methodological and theoretical implications of the findings are discussed. PMID- 12079262 TI - Hereditary pancreatitis: a model for inflammatory diseases of the pancreas. AB - Acute and chronic pancreatitis remain among the most recalcitrant of all diseases to investigation and intervention. In the majority of patients, excessive alcohol consumption is associated with development of the disease. Therefore, several theories have been proposed seeking to explain the relationship between alcohol and the development of acute and chronic pancreatitis. However, recent investigations in hereditary pancreatitis provided important insights into chronic pancreatitis pathogenesis and offer an important model for understanding pancreatic inflammation. This article highlights several advances gained from investigating hereditary pancreatitis kindreds, and reviews the TIGAR-O risk/aetiology classification system. Finally, the major independent theories on development of chronic pancreatitis are reviewed with respect to the SAPE hypothesis of chronic pancreatitis pathogenesis. PMID- 12079263 TI - Clinical relevance of autoimmune-related pancreatitis. AB - Recently, a concept of 'autoimmune pancreatitis' (AIP) was proposed. Computed tomography, magnetic resonance imaging or ultrasonography show a diffusely enlarged pancreas with a so-called 'sausage-like' appearance. Hypergammaglobulinaemia, increased serum levels of total IgG or IgG4, positive autoantibodies such as antinuclear antibody, anti-lactoferrin antibody, anti-CA II antibody, rheumatoid factor and anti-smooth muscle antibody, were often observed in patients with AIP. Microscopic findings showed fibrotic changes with infiltration of lymphocytes, plasmacytes and sometimes eosinophils in the pancreas. Major subgroups of lymphocytes infiltrating areas around pancreatic ducts were CD4(+) T-cells producing IFN-gamma. HLA-DR was expressed on pancreatic duct cells as well as CD4(+) cells. The diagnosis is made by a combination of clinical, laboratory and morphological findings. Laboratory data, pancreas images and diabetes mellitus in most patients do respond to steroid treatment. In conclusion, autoimmune-related pancreatitis appears to be a unique clinical entity. However, its importance in clinical practice needs further characterization. PMID- 12079264 TI - Pancreatic sepsis: prevention and therapy. AB - Except for a minority of early fatalities, most deaths in acute pancreatitis occur after the first 7 to 10 days due to infective complications, particularly infected necrosis. Hence, preventing this risk factor seems to represent a major step forward in the clinical management of severe pancreatitis. Consequently, antibiotics emerged as a cornerstone of the treatment of severe acute pancreatitis. The duration of such treatment, the route of administration and the substance(s) of choice need to be carefully selected. Surgical debridement is the treatment of choice of infected necrosis, while percutaneous drainage is successful in some patients. PMID- 12079265 TI - Enteral and parenteral nutrition in acute pancreatitis. AB - In the last 5 years naso-enteric feeding has increasingly been used in clinical practice in patients with severe acute pancreatitis. Randomized clinical studies in both mild and severe forms of the disease have demonstrated not only the feasibility but also the safety of this approach. The majority of patients have been fed by variously placed nasojejunal tubes with varied problems in maintaining both location and patency. Most have been surprised to find that it is possible to feed the patients in this way with the potential of improving gut barrier function and immune response, at reduced cost and greater safety than with parenteral nutrition. The current evidence points to nasojejunal feeding being preferable to parenteral feeding, but evidence has yet to be produced to prove beyond reasonable doubt that such feeding is an improvement on conservative management without feeding. Finally, the most recent development has indicated that fine-bore nasogastric feeding may well be a realistic alternative to nasojejunal feeding even in the more severe forms of this disease. A small percentage of patients may still need parenteral nutrition. PMID- 12079266 TI - ERCP and MRCP--when and why. AB - Since the introduction of endoscopic retrograde cholangiopancreatography (ERCP) in the 1970s, gastroenterologists have a wide spectrum of diagnostic and therapeutic options in the biliopancreatic ductal system at their disposal. With its arrival in the 1990s, magnetic resonance cholangiopancreatography (MRCP) developed as a potent diagnostic tool in biliopancreatic pathology. Currently, MRCP is widely replacing diagnostic ERCP and thereby avoiding complications related to endoscopic technique.We summarize evidence-based data and demonstrate indications and differential indications for MRCP and ERCP in pancreatic disease. Complications related to the procedures and possible medical prevention are discussed. The feasibility of interventional endoscopy in pancreatic disease is reported in detail. The role of gastroenterologists in performing MRCP is outlined on the basis of practical examples. PMID- 12079267 TI - Genetic basis of pancreatic cancer. AB - Because in the normal state, cells of the pancreas show a very low rate of proliferation, entering the cell cycle is assumed to be the initial event during tumorigenesis. So-called checkpoints monitor cell cycle progression and guarantee the proper duplication of the entire genome. Loss of one or more checkpoints causes subsequent accumulation of genetic alterations which finally results in cancer. Cancer cells are characterized by unrestricted growth, invasion of adjacent tissue and metastasis. All of these features can be explained in terms of genetic changes and the functional consequence of these changes. Activation of the proto-oncogene K-Ras and inactivation of the tumour suppressor gene loci INK4a, p53 and SMAD4 are characteristic for pancreatic cancer. The progression model of pancreatic cancer proposes that pancreatic intraepithelia neoplasia is the pre-cancerous lesion. A preferred genetic pathway has started to evolve. Germ line mutations in specific genes are responsible for cases in which there is a familial predisposition to pancreatic cancer. PMID- 12079268 TI - Surgical and medical therapy for pancreatic carcinoma. AB - Progress on the treatment of pancreatic ductal adenocarcinoma has involved advances in medical and surgical care with important contributions from disciplines such as radiology and intensive care. In the last decade large randomized controlled trials have been undertaken that demonstrate the improved patient outcomes. There is an increased risk of pancreatic cancer in chronic pancreatitis, hereditary pancreatitis and a variety of familial cancer syndromes. The optimum outcome from pancreatic cancer needs management by multidisciplinary teams in regional specialist units. Endoscopic stenting, good pain relief and pancreatic enzyme supplementation are the basis of care in advanced pancreatic cancer. Chemotherapy prolongs survival in advanced pancreatic cancer with little to be gained using drugs other than 5FU. Resection, if possible, prolongs life and provides the best quality of life. Adjuvant chemoradiotherapy is of no benefit but chemotherapy may improve survival. Alongside the evolution in clinical management has been the elucidation of the molecular events that underlie pancreatic cancer and this knowledge has guided the introduction of targeted treatments for pancreatic cancer. PMID- 12079270 TI - New insulins in the treatment of diabetes mellitus. AB - Landmark studies have confirmed the importance of intensified insulin treatment for minimizing long-term diabetic complications. Human insulin is still first line treatment. However, even the most intensive of human insulin-based regimens can only poorly reproduce physiologically desirable insulin release, which includes rapid outbursts of insulin at mealtimes coupled with relatively low and stable basal levels between meals. Encouragingly, there are now four available or soon-to-be-available insulin analogues that offer the potential for more physiological insulin profiles. Insulin lispro and insulin aspart are rapid acting insulin analogues intended for immediate pre-meal administration in type 1 or type 2 diabetes. Compared with injected human insulin, they improve post prandial glucose control and reduce late post-meal and night-time hypoglycaemic episodes. Two basal insulin analogues, insulin glargine and insulin detemir, have also shown beneficial profiles with regard to night-time hypoglycaemia.Some, but not all, studies with the two rapid-acting insulins have shown improvement in overall glucose control, as assessed by HbA(1c), in comparison to human insulin. These results are encouraging and provide hope that entirely analogue-based regimens may improve overall glycaemic control and ease of use of insulin. PMID- 12079271 TI - Use of somatostatin receptor ligands in obesity and diabetic complications. AB - Somatostatin (SMS) is a potent inhibitory molecule. It inhibits both exocrine and endocrine secretory functions of the pancreas, suppresses growth hormone secretion and reduces the level of insulin-like growth factor-1. Long-acting somatostatin analogues were currently investigated for potential clinical benefits in two settings: (a) control of hyperinsulinaemia in obesity and (b) control of an excess of pro-angiogenic factors in diabetes-associated retinal complications. In two randomized, controlled trials the long-acting somatostatin analogue octreotide retarded progression of the microvascular complications in pre-proliferative and advanced stages of diabetic retinopathy. Inhibition of the early phase of insulin secretion by use of octreotide in patients with hypothalamic obesity resulted in weight loss and improved quality of life. Efficacy of octreotide correlated to residual beta-cell activity prior to the treatment. Obesity and diabetes mellitus are the most common chronic metabolic disorders in the world. The use of somatostatin analogues addressing the various hormonal imbalances of these disorders may provide a novel concept for their pharmacological treatment. PMID- 12079272 TI - Cystic fibrosis and related diseases of the pancreas. AB - The discovery of the gene for cystic fibrosis (CF), the cystic fibrosis transmembrane conductance regulator (CFTR), brought about a new era in the study of this disease. Identification of the molecular target has yielded a flood of data that add to our understanding of the pathogenesis, diagnosis and treatment of CF. The CFTR protein is a cAMP-regulated Cl(-) channel with multiple functions in epithelial cells. In the exocrine pancreas the CFTR plays a key role in the apical Cl(-), HCO(3)(-), and water transport in duct cells. The severe loss of functions, caused by mutations of the CFTR gene, leads to pathological lesions of the pancreas. Over 1200 CFTR mutations and polymorphisms have been identified and their diversity may explain the high level of heterogeneity in the CF phenotype. Mutation analyses of the CFTR gene have revealed a spectrum of CFTR-related diseases that do not fit the classical CF picture but are associated with dysfunction of CFTR, such as chronic pancreatitis. PMID- 12079269 TI - Pancreas and islet cell transplantation. AB - Currently, for the patient with type 1 diabetes, a definitive treatment without resorting to the use of exogenous insulin can be achieved only with pancreas or islet cell transplantation. These means of restoring beta-cell mass can completely maintain essentially normal long-term glucose homeostasis, although the need for powerful immunosuppressive regimens limits their application to only a subgroup of adult patients. Apart from the shortage of donors that has limited all kinds of transplantation, however, the widespread use of beta-cell replacement has been precluded until recently by the drawbacks associated with both organ and islet cell transplantation. Although the study of recurrence of diabetes has generated attention, the fundamental obstacle to pancreas and islet transplantation has been, and remains, the alloimmune response. With a better elucidation of the mechanisms of alloengraftment achieved during the last 3 years, the stage has been set for further advances. PMID- 12079274 TI - Gene therapy tools for functional genomics. PMID- 12079275 TI - Minisatellites show rare and simple intra-allelic instability in the mouse germ line. AB - Minisatellites provide very informative systems for analyzing processes of tandem repeat DNA turnover in humans. The mouse genome also contains authentic minisatellites, but none has yet been found to show high levels of instability. Indirect evidence using minisatellite variant repeat mapping by PCR in Mus musculus subspecies suggested that mouse minisatellites mutate at a rate below 10(-3) per gamete and mainly by intra-allelic events. This is in sharp contrast to the complex interallelic mutations observed at high frequency at some human loci. To define more directly the turnover mechanisms and rates of instability at one of the most variable mouse minisatellites (MMS80), we used size-enrichment small-pool PCR (SESP-PCR) to recover de novo mutant alleles from sperm DNA from homozygous BALB/cJ mice and from strain DHA heterozygotes. The sperm mutation rate at MMS80 was extremely low, at or below 5 x 10(-6) per sperm. Comparison of progenitor and mutant allele structures showed that these rare mutants had arisen by simple and primarily, if not exclusively, intra-allelic mutation events. These results suggest a fundamental difference in turnover mechanisms at minisatellites between mice and human. PMID- 12079276 TI - Characterization of FAM10A4, a member of the ST13 tumor suppressor gene family that maps to the 13q14.3 region associated with B-Cell leukemia, multiple myeloma, and prostate cancer. AB - Using the 650-kb DNA sequence from the minimally deleted region in B-cell chronic lymphocytic leukemia (BCLL), we have identified a new gene, FAM10A4, that maps to the proximal end of the region. This gene has been shown to be part of a now six member family of genes with high homology to the ST13 tumor suppressor gene. We have established conditions to specifically undertake mutation studies of the chromosome 13 member of this family and have identified a Ser71Leu change in BCLL samples, which is apparently a polymorphism. The characterization of this gene will permit mutation studies in other tumor cell types such as multiple myeloma and prostate cancer, which also show genetic loss in the 13q14 region. PMID- 12079277 TI - Association of a polymorphism of the CC chemokine receptor-2 gene with bone mineral density. AB - The possible association of the 190G-->A (Val64Ile) polymorphism of the CC chemokine receptor-2 gene (CCR2) with bone mineral density (BMD) was examined in 2215 subjects (1125 men, 1090 women), all of whom were community-dwelling individuals aged 40 to 79 years. Among men aged < 60 years, BMD for the distal radius, lumbar spine, or Ward's triangle was significantly greater in those with the AA genotype than in those with the GG or GA genotypes. For postmenopausal women, BMD for the distal radius or femoral neck was significantly greater in those with the AA genotype than in those with the GG or GA genotypes. In contrast, for men aged > or =60 years and for premenopausal women, BMD was not associated with the CCR2 genotype. These results suggest that CCR2 may be a new candidate for a susceptibility locus for bone mass in middle-aged men and postmenopausal women. PMID- 12079278 TI - A genome scan for Loci associated with aerobic running capacity in rats. AB - Aerobic capacity is a complex trait that defines the efficiency to use atmospheric oxygen as an electron acceptor in energy transfer. Copenhagen (COP) and DA inbred rat strains show a wide difference in a test for aerobic treadmill running and serve as contrasting genetic models for aerobic capacity. A genome scan was carried out on an F(2)(COP x DA) segregating population (n=224) to detect quantitative trait loci (QTLs) associated with aerobic running capacity. Linkage analysis revealed a significant QTL on chromosome 16 (lod score, 4.0). A suggestive linkage was found near the p-terminus of chromosome 3 (lod score, 2.2) with evidence of an interaction with another QTL on chromosome 16 (lod score, 2.9). All three QTLs showed a dominant mode of inheritance in which the presence of at least one DA allele was associated with a greater distance run. These results represent the first aerobic capacity QTLs identified in genetic models. PMID- 12079279 TI - Neuron navigator: a human gene family with homology to unc-53, a cell guidance gene from Caenorhabditis elegans. AB - We have cloned the gene neuron navigator-1 (NAV1), a human homolog of unc-53, a gene involved in axon guidance in Caenorhabditis elegans. Duplications during evolution gave rise to three human homologs located on chromosomes 1q32.1, 11p15.1, and 12q21.1. NAV1 and NAV2 are expressed in the developing brain. NAV1, NAV2, and NAV3 expression is detected in adult heart, kidney, and brain, respectively. NAV1 encodes a protein lacking, in the aminoterminal part, a CH domain present in the other NAV genes. The first exon of NAV1 arose through an ancient internal duplication of sequences that also gave rise to exon 8 of NAV3 and exon 7 of NAV2. A detailed study of the NAV environment on the different chromosomes reveals incomplete micro-syntheny between the three regions. Through analysis of the phylogenetic relationships for three different gene families in the NAV environment, we reconstructed part of the events that formed these regions. PMID- 12079280 TI - The mouse Laf4 gene: exon/intron organization, cDNA sequence, alternative splicing, and expression during central nervous system development. AB - The cerebral cortex is a tissue with a high degree of neuronal diversity. It consists of six cell layers with a unique set of neuronal subtypes. A crucial step in the process of cortical differentiation is the transition from a mitotically active neuroblast to a postmitotic young neuron. To identify genes involved in the control of this transition, we applied a novel method of cDNA subtraction based on mirror-orientation selection. One of the genes we have identified in our screening proved to be a mouse homolog of the human putative transcription factor LAF4. We identified alternatively spliced forms of mouse Laf4 that encode several forms of putative protein with potentially different transactivation functions. Two forms are expressed mainly during embryogenesis, whereas the other forms are expressed mainly in adults. We have found that Laf4 transcription becomes very quickly upregulated as soon as young cortical neurons leave the ventricular zone (VZ), the cortical-proliferative compartment. This coincides with the initial steps of cortical differentiation. Laf4 becomes downregulated in postnatal cortex, indicating its involvement in the transcriptional regulation of the early steps of cortical differentiation. We have also examined Laf4 expression in the brains of Sey and reeler mutants. Laf4 was downregulated in the lateroventral part of the cerebral cortex and completely lost in the piriform cortex of the Sey mutant embryos. We also compared its expression during central nervous system development with that of its closest homolog, Fmr2, a gene implicated in mental retardation in humans. PMID- 12079281 TI - The gamma S-crystallin gene is mutated in autosomal recessive cataract in mouse. AB - We established a recessive cataract model from a spontaneous mutation in the KUNMING outbred mice. Lens opacity appears 11 days after birth. Slit lamp examination reveals that the opacity mainly localizes to the nuclear region of the lens. Histological analysis shows a severe degeneration of the epithelial cells underneath the anterior lens capsule, whereas those cells in the equatorial region display an excessive proliferation and migration. Within the cortical area underneath the posterior lens capsule, both vacuoles and morgagnian-like bodies are seen. Blue-stained spherical bodies are observed in the embryonic nucleus, forming a Y-like pattern. We mapped the disease locus and found a homozygous G to A nucleotide conversion at position 489 of Crygs in mutant mice, leading to a truncated gene product (Trp163Stop). This finding suggests that CRYGS is not only a lens structural protein, but is also likely to be involved in epithelial cell proliferation, apoptosis, and migration. PMID- 12079282 TI - Novel vertebrate genes and putative regulatory elements identified at kidney disease and NR2E1/fierce loci. AB - Fierce (frc) mice are deleted for nuclear receptor 2e1 (Nr2e1), and exhibit cerebral hypoplasia, blindness, and extreme aggression. To characterize the Nr2e1 locus, which may also contain the mouse kidney disease (kd) allele, we compared sequence from human, mouse, and the puffer fish Fugu rubripes. We identified a novel gene, c222389, containing conserved elements in noncoding regions. We also discovered a novel vertebrate gene conserved across its length in prokaryotes and invertebrates. Based on a dramatic upregulation in lactating breast, we named this gene lactation elevated-1 (LACE1). Two separate 100-bp elements within the first NR2E1 intron were virtually identical between the three species, despite an estimated 450 million years of divergent evolution. These elements represent strong candidates for functional NR2E1 regulatory elements in vertebrates. A high degree of conservation across NR2E1 combined with a lack of interspersed repeats suggests that an array of regulatory elements embedded within the gene is required for proper gene expression. PMID- 12079283 TI - The zinc-finger transcription factor INSM1 is expressed during embryo development and interacts with the Cbl-associated protein. AB - Here we describe the isolation and characterization of the mouse homolog of the human zinc-finger transcription factor INSM1 (IA-1) and identify an interacting protein. A 2.9-kb cDNA with an open reading frame of 1563 nucleotides, corresponding to a translated protein of 521 amino acids, was isolated from a mouse beta TC-1 cDNA library. Mouse INSM1 was found to be 86% identical to human INSM1 and both proteins contain proline-rich regions and multiple zinc-finger DNA binding motifs. Sequencing of mouse Insm1 genomic DNA revealed that it is an intronless gene. Chromosomal mapping localized Insm1 to chromosome 2. Northern blot analysis showed that mouse Insm1 expression begins at 10.5 days in the embryo, decreases after 13.5 days, and is barely detected at 18.5 days. In mouse brain, Insm1 is strongly expressed for 2 weeks after birth but shows little or no expression thereafter. Transfection of cells with GFP-tagged INSM1 revealed that INSM1 is expressed exclusively in the nucleus. We identified proteins that interacted with INSM1 by the yeast two-hybrid system and the binding of one of them, Cbl-associated protein (CAP), to INSM1 was confirmed by in vitro pull-down experiments, nuclear colocalization, and co-immunoprecipitation assays. Further studies showed that both INSM1 and CAP proteins were present in the nucleus of insulinoma cells and that endogenous INSM1 protein was co-precipitated with antibody to CAP. These findings raise the possibility that during embryo development CAP may enter the nucleus through its own nuclear localization signal or by binding to INSM1. PMID- 12079284 TI - Molecular cloning and characterization of human chondrolectin, a novel type I transmembrane protein homologous to C-type lectins. AB - CHODL, a novel human gene encoding chondrolectin, was isolated by PCR screening. It is localized at chromosome 21q21 and consists of six exons and five introns. The open reading frame of CHODL encodes a type I transmembrane protein containing a single carbohydrate recognition domain (CRD) of C-type lectins in its extracellular portion. CHODL was detected as a 2.6-kb transcript by northern blot using enriched human testis RNA. RT-PCR analysis revealed preferential expression of CHODL in testis, prostate, and spleen. Immunohistochemistry demonstrated that the expression of CHODL is mainly limited to vascular muscle of testis, smooth muscle of prostate stroma, heart muscle, skeletal muscle, crypts of small intestine, and red pulp of spleen. Western blot analysis revealed that CHODL is an N-glycosylated protein with a molecular weight of approximately 36 kDa. In transiently transfected COS1 cells, CHODL shows a predominantly perinuclear localization. Although the predicted CHODL protein shares a significant homology (45% overall and 60% within the CRD) with layilin, a recently identified hyaluronan receptor, we failed to detect a specific interaction between CHODL and hyaluronan using cetylpyridinium chloride precipitation. PMID- 12079285 TI - Structure and chromosomal distribution of human mitochondrial pseudogenes. AB - Nuclear mitochondrial pseudogenes (Numts) have been found in the genome of many eukaryote species, including humans. Using a BLAST approach, we found 1105 DNA sequences homologous to mitochondrial DNA (mtDNA) in the August 2001 Goldenpath human genome database. We assembled these sequences manually into 286 pseudogenes on the basis of single insertion events and constructed a chromosomal map of these Numts. Some pseudogenes appeared highly modified, containing inversions, deletions, duplications, and displaced sequences. In the case of four randomly selected Numts, we used PCR tests on cells lacking mtDNA to ensure that our technique was free from genome-sequencing artifacts. Furthermore, phylogenetic investigation suggested that one Numt, apparently inserted into the nuclear genome 25-30 million years ago, had been duplicated at least 10 times in various chromosomes during the course of evolution. Thus, these pseudogenes should be very useful in the study of ancient mtDNA and nuclear genome evolution. PMID- 12079286 TI - A novel, evolutionarily conserved gene family with putative sequence-specific single-stranded DNA-binding activity. AB - Complete and partial deletions of chromosome 5q are recurrent cytogenetic anomalies associated with aggressive myeloid malignancies. Earlier, we identified an approximately 1.5-Mb region of loss at 5q13.3 between the loci D5S672 and D5S620 in primary leukemic blasts. A leukemic cell line, ML3, is diploid for all of chromosome 5, except for an inversion-coupled translocation within the D5S672 D5S620 interval. Here, we report the development of a bacterial artificial chromosome (BAC) contig to define the breakpoint and the identification of a novel gene SSBP2, the target of disruption in ML3 cells. A preliminary evaluation of SSBP2 as a tumor suppressor gene in primary leukemic blasts and cell lines suggests that the remaining allele does not undergo intragenic mutations. SSBP2 is one of three members of a closely related, evolutionarily conserved, and ubiquitously expressed gene family. SSBP3 is the human ortholog of a chicken gene, CSDP, that encodes a sequence-specific single-stranded DNA-binding protein. SSBP3 localizes to chromosome 1p31.3, and the third member, SSBP4, maps to chromosome 19p13.1. Chromosomal localization and the putative single-stranded DNA binding activity suggest that all three members of this family are capable of potential tumor suppressor activity by gene dosage or other epigenetic mechanisms. PMID- 12079287 TI - Transcription regulation of human chemokine receptor CCR3: evidence for a rare TATA-less promoter structure conserved between drosophila and humans. AB - The chemokine receptor CCR3 has a critical function in the pathogenesis of eosinophilic diseases and is an entry co-receptor for HIV-1. We describe here the genomic organization and general transcriptional control mechanism for the human gene CCR3. We identified six cDNA transcripts formed by alternative splicing of eight exons and seven introns. CCR3 contains a 37-bp core promoter domain (-3 to +34 relative to the transcription start point) lacking a TATA box but inclusive of an initiator sequence, a G at +24, and a downstream promoter element (DPE) at +28 to +33 common for Drosophila melanogaster but heretofore described for only two other human genes. Mutation of these elements significantly attenuates CCR3 transcription, as predicted by a model of RNA pol II engagement with DPE containing Drosophila promoters. These results provide evidence for the functional conservation of a DPE-dependent, general transcription control mechanism between Drosophila and human genes. PMID- 12079288 TI - A novel gene encoding a TIG multiple domain protein is a positional candidate for autosomal recessive polycystic kidney disease. AB - Autosomal recessive polycystic kidney disease (ARPKD) is a common hereditary renal cystic disease in infants and children. By genetic linkage analyses, the gene responsible for this disease, termed polycystic kidney and hepatic disease 1 (PKHD1), was mapped on human chromosome 6p21.1-p12, and has been further localized to a 1-cM genetic interval flanked by the D6S1714/D6S243 (telomeric) and D6S1024 (centromeric) markers. We recently identified a novel gene in this genetic interval from kidney cDNA, using cloning strategies. The gene PKHD1 (PKHD1-tentative) encodes a novel 3396-amino-acid protein with no apparent homology with any known proteins. We named its gene product "tigmin" because it contains multiple TIG domains, which usually are seen in proteins containing immunoglobulin-like folds. PKHD1 encodes an 11.6-kb transcript and is composed of 61 exons spanning an approximately 365-kb genomic region on chromosome 6p12-p11.2 adjacent to the marker D6S1714. Northern blot analyses demonstrated that the gene has discrete bands with one peak signal at approximately 11 kb, indicating that PKHD1 is likely to have multiple alternative transcripts. PKHD1 is highly expressed in adult and infant kidneys and weakly expressed in liver in northern blot analysis. This expression pattern parallels the tissue involvement observed in ARPKD. In situ hybridization analysis further revealed that the expression of PKHD1 in the kidney is mainly localized to the epithelial cells of the collecting duct, the specific tubular segment involved in cyst formation in ARPKD. These features of PKHD1 make it a strong positional candidate gene for ARPKD. PMID- 12079289 TI - Canine PKD1 is a single-copy gene: genomic organization and comparative analysis. AB - Most cases of autosomal dominant polycystic kidney disease are caused by mutations in the gene PKD1, encoding polycystin-1. To gain insight into the role of polycystin-1 in tubulogenesis and cystogenesis using the well-characterized canine kidney epithelial cell line MDCK, we have now cloned and characterized the exon/intron structure of the canine gene PKD1. FISH analysis showed that the dog genome lacks the multiple PKD1 homologs present in human. Intron 21 of dog PKD1 lacked the polypyrimidine tract characteristic of the human gene, whereas pyrimidine-rich elements were identified in canine intron 30. Canine polycystin-1 showed a higher degree of homology with the human counterpart and lower homology with mouse and rat. A striking degree of conservation (97% identity) was determined for the leucine-rich repeat domain between dog and human. Also, the homology analysis indicated that 4 of 16 Ig-like repeats (IgIII, IgVII, IgX, and IgXV) are likely to be functionally significant. This is particularly important in light of our recent findings demonstrating that Iglike domains form strong homophilic interactions and can mediate cell-cell adhesion. These data enable detailed analysis of the role of polycystin-1 in cystogenesis and tubulogenesis using the canine MDCK cell line. PMID- 12079290 TI - Transcriptional analysis of a novel cluster of LY-6 family members in the human and mouse major histocompatibility complex: five genes with many splice forms. AB - Lymphocyte antigen-6 (LY-6) superfamily members are cysteine-rich, generally GPI anchored cell surface proteins, which have definite or putative immune related roles. A cluster of five potential LY-6 superfamily members is located in the human and mouse major histocompatibility complex class III region. Comparative analysis of their genomic and cDNA sequences allowed us to carry out detailed annotations of these genes. We analyzed their mRNA expression patterns by RT-PCR performed on human and mouse cell line and tissue RNA. Sequence analysis of the transcripts revealed splice variants of all these genes in humans, and all but one in mouse. These splice forms retained introns or intron fragments, mainly generating premature stop codons, such that the only potentially functional mRNA was the predicted form. In some cases, the mis-spliced form was the most abundant form, suggesting a control mechanism for gene expression. Each gene showed mRNA expression differences between human and mouse. PMID- 12079291 TI - Estrogen and cancer. PMID- 12079292 TI - Delivery outcomes following loop electrosurgical excision procedure for microinvasive (FIGO stage IA1) cervical cancer. AB - OBJECTIVE: The goal of this study was to report the delivery outcomes in women who had loop electrosurgical excision procedure (LEEP) for microinvasive cervical cancer (stage IA1 without lymphovascular invasion) and became pregnant and progressed beyond 24 weeks. METHODS: A case-control study was performed. Twenty eight women who were managed exclusively with LEEP for microinvasive cervical carcinoma had at least one pregnancy beyond 24 weeks and were the cases. Each case was matched with one woman who delivered at the same department without prior treatment of her cervix (controls) and their delivery outcomes were compared with those of the cases. Known risk factors for preterm delivery were used as matching factors. RESULTS: There was no statistically significant difference (P > 0.05) between cases and controls in the duration of pregnancy (37.6-38.4 weeks respectively), birth weight (3212-3315 g), cesarean section rate (17.8-32.1%), neonatal unit admission rate (21.4-10.7%), and precipitate labor rate (13-10.5%). The duration of labor was significantly shorter in cases (5.5 7.1 h, P = 0.032). After LEEP the relative risk for preterm delivery is 3.67 (95% confidence interval, 0.97-20.27), for low birth weight infant 0.67 (0.06-5.8), for precipitate labor 1 (0.05-3.88), and for delivery by cesarean section 0.5 (0.17-4.46). CONCLUSIONS: Women treated for microinvasive cancer with LEEP did not have significantly more delivery complications compared with controls apart from shorter duration of labor. There was a possible non-statistically significant trend toward shorter duration of pregnancy in cases. While caution should be advised when selecting and treating women with microinvasive carcinoma by LEEP, the apparent safety of the management and the satisfactory delivery outcome seem to justify this approach in many cases. PMID- 12079293 TI - Apotosis-related proteins in cervical intraepithelial neoplasia and squamous cell carcinoma of the cervix. AB - OBJECTIVES: A loss of balance between proliferation and apoptosis leads to tumor formation. Normal cervical epithelium becomes dysplastic before potentially developing into carcinoma. This study was conducted to delineate the role of apoptosis-related proteins in various stages of development in cervical neoplasia. Both regulator and effector proteins were examined. METHODS: The expression of apoptosis-related proteins, including five members of the Bcl-2 family and two members of the caspase family, was evaluated in 26 low-grade cervical intraepithelial neoplasias (CIN-L), 37 high-grade cervical intraepithelial neoplasias (CIN-H), and 43 cervical squamous cell carcinomas, using immunohistochemistry. Specimens showing cytosolic immunoreactivity in 10% or more of the neoplastic cells were considered immunopositive. RESULTS: One hundred six subjects were studied. All seven apoptotic regulators examined were positive in a proportion of these neoplasms. The expression of Caspase 3 was significantly higher in CIN-H than in CIN-L (P = 0.016). The expression of Bak, Caspase 3, and Caspase 6 was reduced in cervical carcinoma compared to CIN-H (P < 0.01, P = 0.026, P < 0.01, respectively). CONCLUSIONS: There is an increase in expression of Caspase 3 in CIN-H compared to CIN-L and the increase is thought to be related to the increased proliferative activity in dysplastic cells. The reduction of Bax, Caspase 3, and Caspase 6 expression in carcinoma indicates that the apoptotic mechanism has become defective in the process of malignant transformation. PMID- 12079294 TI - Overexpression of a stabilized mutant form of the cyclin-dependent kinase inhibitor p27(Kip1) inhibits cell growth. AB - OBJECTIVE: The purpose of this study was to test the hypothesis that the expression of the mutant p27(Kip1) protein enhances cell growth inhibition and is more stable than that of the wild-type p27(Kip1). METHODS: Site-directed mutagenesis was used to mutate threonine 187 to an alanine residue, generating the mutant p27(Kip1). To study the effects of the p27(Kip1) mutant on cell growth, luciferase assays were performed. Cells were transiently transfected with the Renilla luciferase reporter construct and empty vector, wild-type p27(Kip1), or mutant p27(Kip1) using Fugene 6. The transfected cells were lysed and assayed for luciferase activity 24 h later with a dual-luciferase reporter assay system. To further assess the effects of the p27(Kip1) mutant on cell growth, colony count assays were performed. The experiments were repeated in duplicate and a standard two-tailed Student t test was use to analyze the data. RESULTS: Wild type p27(Kip1) protein has a half-life of approximately 2 h while the p27(Kip1) mutant has a half-life of greater than 12 h. Furthermore, the p27(Kip1) mutant retained the ability to inhibit CDK2-associated H1 kinase activity. Cells expressing the p27(Kip1) mutant had an 88% reduction in luciferase activity compared to cells expressing the wild-type p27(Kip1) (P = 0.001). Colony assays revealed that cells expressing the p27(Kip1) mutant had fewer colonies compared to cells expressing the wild-type p27(Kip1) (P = 0.04). CONCLUSIONS: These data are consistent with the hypothesis that the mutated form of p27(Kip1) is more effective in cell growth inhibition than the wild-type p27(Kip1) protein. PMID- 12079296 TI - Can a preoperative CA 125 level be a criterion for full pelvic lymphadenectomy in surgical staging of endometrial cancer? AB - OBJECTIVE: The aim of the present study was to evaluate whether a preoperative serum level of CA 125 in patients with endometrial cancer can provide additional information in determining the extent of lymphadenectomy required in the surgical staging and which cutoff value is optimal in this respect. METHODS: CA 125 was measured in 124 patients diagnosed with endometrial carcinoma who underwent surgery at our institution between January 1995 and May 2000. Statistic analysis was performed using the Mann-Whitney U test and Kruskal-Wallis test to evaluate the association of preoperative CA 125 levels with various factors. The chi(2)/Fisher's exact test and a logistic regression model were employed to examine the effects of clinicopathological factors on serum CA 125 levels. Furthermore, a receiver operating characteristic curve was used to determine which cutoff value of the preoperative CA 125 was the optimal one. RESULTS: Univariate analyses showed that elevated CA 125 levels were significantly correlated with an advanced stage, larger tumor size, increasing depth of the myometrial invasion, cervical invasion, positive cytology, and lymph node metastases (P < 0.05). Multivariate analyses using a logistic regression model showed lymph node metastases had the most significant effect on the elevation of CA 125 levels. The ROC curve determined that the best cutoff value was 40 U/ml; the sensitivity and specificity for screening lymph node metastases were found to be 77.8 and 81.0%, respectively. CONCLUSION: Our data provide evidence indicating that a preoperative CA 125 level greater than 40 U/ml can be considered a criterion for full pelvic lymphadenectomy in the surgical staging of endometrial cancer. PMID- 12079295 TI - Effect of fenretinide on ovarian carcinoma occurrence. AB - OBJECTIVE: The aim of this study was to update the effect of fenretinide, a synthetic vitamin A analogue proposed for chemoprevention, on the occurrence of ovarian carcinoma. METHODS: Data were obtained from a randomized clinical trial for the prevention of second breast cancer. For the present investigation, events of interest were new primary carcinomas of the ovary arising in the fenretinide or the no-treatment (control) arm. The probability of carrying a BRCA germ-line mutation was assessed in women with ovarian carcinoma according to G. Parmigiani et al. (1998, Am J Hum Genet 62, 145-58). RESULTS: Fenretinide reduced ovarian carcinoma occurrence during the 5-year intervention period (0 versus 6 cases in the fenretinide and control arm, P = 0.0327). This effect was no longer evident after the 5-year intervention period (6 versus 4 cases, P = 0.7563). Therefore with median observation time of 121 months, a total of 6 carcinomas of the ovary occurred in the fenretinide group and 10 in the control group. The probability of carrying a BRCA mutation was lower for women with ovarian carcinoma in the treatment arm. CONCLUSION: Fenretinide treatment was associated with a lower incidence of ovarian carcinoma during the intervention period but such a protective effect seems to disappear after treatment. Furthermore, a possible protective effect of fenretinide in BRCA-mutated women was suggested. Further studies on fenretinide for the prevention of ovarian carcinoma particularly in women with genetic susceptibility appear necessary. PMID- 12079297 TI - Long-term survival and patterns of care in women with ovarian tumors of low malignant potential. AB - OBJECTIVES: The objectives of this study were to ascertain long-term survival and patterns of care among women diagnosed with ovarian tumors of low malignant potential (LMP) in a population-based data set. METHODS: Using the NCI's Surveillance, Epidemiology, and End Results (SEER) database, we identified 2818 women diagnosed with ovarian tumors of low malignant potential between 1988 and 1997. RESULTS: By FIGO stage, 10-year relative survival was as follows: stage I, 99%; stage II, 98%; stage III, 96%; and stage IV 77%. One-quarter of women with stage I disease underwent partial or unilateral oophorectomy only, while women with more advanced disease commonly underwent omentectomy, unilateral or bilateral oophorectomy, and hysterectomy. Adjuvant chemotherapy was given to about 30% of women with stage III and IV disease. Radiation therapy was rarely used. We observed no significant changes in primary surgery or adjuvant treatment over time. CONCLUSIONS: The diagnosis of an ovarian tumor of LMP conveys a relatively benign prognosis. Conservative surgery should be considered in younger women with early-stage disease. There are insufficient data to support a role for adjuvant chemotherapy for women with advanced disease. PMID- 12079298 TI - Assessment of prognostic factors in stage IIIA endometrial cancer. AB - OBJECTIVE: The objective of this study was the assessment of prognostic factors in stage IIIA endometrial cancer. METHODS: Between 1984 and 1993, 51 patients with stage IIIA endometrial cancer received definitive treatment at our institution. Thirty-seven patients had positive peritoneal cytologic findings only (stage IIIA1), and 14 had adnexal or uterine serosal involvement (USI) (stage IIIA2). Median follow-up of surviving patients was 82.5 months. RESULTS: The 5-year disease-related survival (DRS) and recurrence-free survival (RFS) were 88 and 73%, respectively. RFS was 79% in patients with stage IIIA1 disease, compared with 57% in patients with stage IIIA2 disease (P = 0.04). However, DRS did not significantly differ between stages IIIA1 and IIIA2. In the 37 patients with stage IIIA1 tumors, histologic grade 3, nonendometrioid histologic subtype, and lymphovascular invasion (LVI) significantly predicted a poor prognosis, with extraabdominal sites of failure (P < 0.05). Of the 22 patients who had stage IIIA1 disease with endometrioid histologic subtype and without LVI, none had recurrence [17 had whole abdominal irradiation (WAR) or intraperitoneal injection of (32)P, 2 had pelvic external radiotherapy (PRT)]. By contrast, of the 15 patients with either nonendometrioid histologic subtype or LVI, 9 (60%) had recurrence and 7 (47%) died of disease (12 had WAR or (32)P). An extraabdominal component was present in 7 of the 9 recurrences observed in this subgroup. Among the 14 patients with stage IIIA2 tumors (6 had WAR, 6 had PRT), those with USI had a 5-year DRS of 83% and a rate of extraabdominal failure of 83%, compared with 100 and 12.5% in patients without USI (P < 0.05). CONCLUSION: Patients with stage IIIA endometrial cancer who have endometrioid tumors, no LVI, and positive peritoneal cytologic findings as the only sign of extrauterine disease have an excellent prognosis. Nonendometrioid histologic subtype, LVI, and USI are strong predictors of distant failures and poor prognosis. Patients with either of these histologic factors should be considered candidates for systemic adjuvant therapy. PMID- 12079299 TI - Expression of sialyltransferase family members in cervix squamous cell carcinoma correlates with lymph node metastasis. AB - OBJECTIVE: Altered messenger ribonucleic acid (mRNA) expression of the four sialyltransferases (STs including ST3Gal I, ST3Gal III, ST3Gal IV, and ST6Gal I) is important in squamous cell carcinoma of the cervix. This study further investigates their changes in mRNA expression of the four STs in FIGO stage IB1 squamous cell carcinoma to assess the extent of sialylation associated with lymph node metastases. METHODS: Alterations in ST mRNA expression in FIGO IB1 cervical squamous cell carcinomas (n = 79) were examined by semiquantitative reverse transcription-polymerase chain reaction. RESULTS: Both ST6Gal I mRNA and ST3Gal III mRNA expressions were significantly increased in patients with lymph node metastases compared to those without lymph node metastases (P = 0.002 and P = 0.001, respectively, Mann-Whitney U test). Using receiver operating characteristic curves of ST ratio index for accuracy comparison of lymph node metastases, ST3Gal III and ST6Gal I were observed to be fairly interchangeable (area under the curve (AUC) of 3Gal I = 0.810; AUC of 6Gal I = 0.786, significance of difference between AUC = 0.810). High ST6Gal I expression was associated with other invasive properties of cervical cancer, such as deep stromal invasion and presence of lymph-vascular space involvement. ST6Gal I expression seemed to be more enhanced in bigger tumors. CONCLUSIONS: Our results suggested that ST3Gal III and ST6Gal I were of importance for the lymph node metastases in FIGO IB1 cervical cancer patients; more specifically, overexpression of ST6Gal I was of crucial relevance for the presence of poor prognostic factors, such as deep stromal invasion and lymph-vascular space involvement and lymph node metastases. PMID- 12079300 TI - Detection of beta-subunit human chorionic gonadotropin mRNA in the peripheral blood of patients with nonmetastatic gestational trophoblastic disease. AB - OBJECTIVE: The aim of this study was to detect beta-subunit human chorionic gonadotropin (beta(h)CG) mRNA in the peripheral blood samples of patients with nonmetastatic gestational trophoblastic disease (GTD) undergoing hysterectomy. METHODS: Heparinized peripheral blood samples were obtained from four patients with nonmetastatic GTD before, during, and after hysterectomy. The beta(h)CG mRNA expression was examined by reverse transcriptase-polymerase chain reaction using beta(h)CG primers. The expression of beta(h)CG mRNA was quantified using a densitometer. RESULTS: Beta(h)CG expression was detected in all patients before and during hysterectomy. The expression of beta(h)CG mRNA during operation was so high that it could not be quantified using densitometer. The expression decreased rapidly after operation. CONCLUSIONS: Disseminated trophoblastic cells are present in the peripheral blood even in cases without metastasis. Trophoblastic cells circulating in the peripheral blood can be reduced by surgical intervention. PMID- 12079301 TI - Can serum CA-125 levels predict the optimal primary cytoreduction in patients with advanced ovarian carcinoma? AB - OBJECTIVE: The aim of this study was to investigate the predictive value of serum CA-125 levels to ability of optimal primary cytoreduction in patients with advanced epithelial ovarian carcinoma. METHODS: Preoperative serum CA-125 levels were determined by a commercial enzyme immunoassay kit in a series of 92 patients with stage IIIC epithelial ovarian carcinoma. The abilities of various cutoff value of CA-125 to predict suboptimal cytoreductive surgery were determined. A receiver operating characteristic curve was used to find the most clinically useful CA-125 cutoff value. RESULTS: Optimal cytoreduction was obtained in 48 patients (52%) using the diameter of the largest residual tumor nodule less than 1 cm. Receiver operating characteristic curve showed that the most clinically suitable CA-125 cutoff value was 500 U/ml. Forty-seven patients (51%) had preoperative serum CA-125 levels below 500 U/ml. Of these patients, optimal cytoreductive surgery was performed in 36 (77%). Of the 45 patients with serum CA 125 levels greater than 500 U/ml, optimal cytoreductive surgery was achieved in 12 (27%). True- and false-positive rates were 73 and 23%, respectively. CONCLUSIONS: Although our results showed that preoperative serum CA-125 levels might predict the optimal resectable patients, larger prospective studies are needed to prove its predictivity. Gynecologic oncologists should evaluate the sum of all criteria until more data are available. PMID- 12079302 TI - Microsatellite instability, MLH1 promoter methylation, and loss of mismatch repair in endometrial cancer and concomitant atypical hyperplasia. AB - OBJECTIVE: MLH1 methylation is associated with the microsatellite instability (MSI) phenotype in endometrial cancer and atypical endometrial hyperplasia, a premalignant precursor to carcinoma. The observation that methylation is also seen in atypical endometrial hyperplasia without MSI suggests that methylation is an early event in endometrial tumorigenesis. Our objective was to determine if methylation is always present in MSI-positive atypical hyperplasia concomitant with MSI-positive, methylation-positive carcinoma. METHODS: We used laser capture microdissection to study MLH1 methylation and MSI in a large series of endometrial cancer cases that had previously been shown to have methylation and the MSI-high (MSI-H) phenotype. We resampled areas of carcinoma from 27 patients along with 51 foci of concomitant atypical endometrial hyperplasia. RESULTS: Consistent with previous reports, we saw MLH1 methylation in areas of atypical endometrial hyperplasia that did not show MSI. In addition, we noted that 18% of the MSI-H atypical endometrial hyperplasia DNAs lacked methylation of critical cytosines in the MLH1 promoter. Immunohistochemistry studies showed that these MSI-H unmethylated foci of atypical endometrial hyperplasia failed to express MLH1, as did regions of simple hyperplasia. CONCLUSION: Methylation of the MLH1 promoter is an early event in endometrial tumorigenesis. Given that not all MSI positive tissues had methylation at cytosines -229 and -231, it appears that methylation may not be required for MLH1 silencing and loss of mismatch repair. PMID- 12079303 TI - Loss of expression and altered localization of KAI1 and CD9 protein are associated with epithelial ovarian cancer progression. AB - OBJECTIVE: Impairment of cell adhesion plays a vital role in tumor progression. E and N-cadherin, CD9, and KAI1 are all adhesion molecules that have been implicated in the progression of several different tumor types. To help explain the potential role these adhesion molecules have in ovarian cancer, comparisons were made between expression patterns in normal ovary and various grades of primary and metastatic epithelial ovarian cancers. METHODS: Thirty-two primary and 8 metastatic human ovarian epithelial carcinomas and 18 samples of normal ovarian tissue were examined for adhesion molecule expression using immunohistochemistry. RESULTS: KAI1 and CD9 revealed an inverse relationship between tumor grade and expression levels, characterized by high expression in low-grade tumors and low expression in high-grade tumors and metastases. KAI1 and CD9 also demonstrated a shift in cellular localization from the membrane in grade 1 tumors to the cytoplasm in grade 3 tumors. N-cadherin expression showed a positive trend between expression levels and tumor grade. E-cadherin expression varied little between different tumor grades and metastases. Inclusion cysts (n = 6) and surface invaginations often strongly expressed KAI1, CD9, and E-cadherin. KAI1 expression was variable in ovarian follicles and corpora lutea depending on their stage of development. CONCLUSIONS: Although sample size is limited, these findings suggest that progression of ovarian epithelial carcinomas is associated with down-regulation and altered cellular localization of KAI1 and CD9. In addition, variable KAI1 expression during follicular and luteal development suggests that it has a physiological function in the ovary. Further investigation will be needed to see if it is also regulated this way during progression of ovarian cancers. PMID- 12079304 TI - The significance of pneumatosis intestinalis or bowel perforation in patients with gynecologic malignancies. AB - OBJECTIVES: The objectives of this study were to evaluate the clinical significance and outcome of pneumatosis intestinalis or bowel perforation in patients with gynecologic malignancies. METHODS: A retrospective review of all gynecologic oncology patients at our institution from 1996 to the present was performed to identify computed tomography examinations showing pneumatosis, free air, or the presence of portal venous gas. Admission symptoms, laboratory testing, radiographic and operative findings, and overall survival were evaluated. At the discretion of the attending gynecologic oncologist, patients were managed either surgically or conservatively. Statistical analysis was performed with Fisher's exact test. RESULTS: Twenty-eight patients met study criteria. Thirteen patients were excluded as a result of radiographic free air immediately following surgery, thus leaving 15 patients for analysis. Sixty percent (n = 9) of patients were managed surgically while 40% (n = 6) were managed conservatively. Pain and tenderness at presentation prompted surgical intervention in a statistically significant number of patients (P = 0.04). No other sign or symptom was significant. Of the 6 patients managed conservatively, 3 (50%) died within 1 week of diagnosis. Survival for the others was 2, 4, and 6 months. Of the 9 patients managed surgically 6 (67%) patients died, 4 within 2 weeks of surgery and the remainder at 2 and 12 months postoperatively. The 3 patients who survived all had surgical intervention and none had radiographic or pathologic evidence of cancer at the time of presentation (P = 0.01). Overall mortality at 6 months was 73% (11/15). Surgical management was associated with prolonged ICU care, mechanical ventilation, and sepsis. CONCLUSIONS: Pneumatosis intestinalis and bowel perforation carry a grave prognosis for patients with gynecologic malignancies. These data suggest that patients explored for radiographic evidence of pneumatosis or perforation with preoperative evidence of active malignancy do not survive the immediate postoperative period. The balance between quality and quantity of life must be considered when weighing the options for the management of this condition. PMID- 12079306 TI - A case report of recurrent epithelial ovarian cancer metastatic to the sternum, diaphragm, costae, and bowel managed by aggressive secondary cytoreductive surgery without postoperative chemotherapy. AB - BACKGROUND: Ovarian epithelial cancer typically presents in advanced stage and has been traditionally managed by a combination of cytoreductive surgery followed by adjuvant systematic chemotherapy. The management of recurrent ovarian cancer has been individualized: surgical resection of intraabdominal and/or pelvic disease has been performed when technically feasible and usually followed with chemotherapy. CASE: This case describes aggressive surgical management of recurrent ovarian cancer metastatic to the lower ribs, sternum, and diaphragm. A clear cell, Stage IIIA ovarian cancer was successfully resected in a 73-year-old female. The patient had total abdominal hysterectomy, bilateral salpingo oophorectomy, and omentectomy followed by six cycles of adjuvant chemotherapy, consisting of cyclophospamide and carboplatinum. A period of 8 years elapsed before recurrent disease was detected; there were two separate metastatic sites. A secondary cytoreductive surgery without further chemotherapy has been the mainstay of treatment. A combination of exploratory laparotomy and en bloc resection revealed the metastatic deposits, a 5-cm mass involving the diaphragm, the lower aspect of the manubrium sternum, and four right lower ribs. The second deposit was identified in the left paracolic gutter invading the sigmoid colon. CONCLUSION: At 47 months of follow-up, the patient is alive and without any evidence of measurable disease by exam and confirmed by CT scans of chest, abdomen, and pelvis. To our knowledge, this is one of the few reported cases managed successfully by surgical approach and is recommended in selected patients with metastatic ovarian cancer. PMID- 12079305 TI - Ovarian cancer p53 mutation is associated with tumor microvessel density. AB - OBJECTIVE: The objective of this study was to investigate the relationship between microvessel density, as measured by CD31 staining, and histopathologic factors as well as p53 tumor suppressor gene mutation in ovarian cancer. METHODS: Ovarian cancers (n = 77) were analyzed for p53 gene mutations and CD31 immunohistochemical expression. Histopathologic and mutational data were related to CD31 staining utilizing the Mantel correlation statistic. The microvessel density was scored by averaging counts from three high-power (200x) fields. Survival was based upon maximizing the hazard ratio. RESULTS: The mean microvessel density counts based on CD31 staining (vessels/HPF) for each FIGO stage and mutation type are as follows: Stage I (10.2), Stage II (10.7), Stage III (13.8), Stage IV (22.0), wild-type p53 (9.3), missense p53 mutation (14.4), and null p53 mutation (23.1). There was a significant correlation between microvessel density count and FIGO stage (P = 0.026), grade (P = 0.04), and p53 mutation type (P = 0.02). Median survival was more than doubled (6.4 vs 2.9 years; P = 0.009) for tumors with microvessel density counts less than or equal to 14 vessels/HPF. CONCLUSIONS: These data are consistent with the hypothesis that ovarian cancer p53 mutation functions to directly influence angiogenesis, which in turn compromises disease-specific survival. It also suggests validity to targeting p53 alterations with gene replacement as well as the use of antiangiogenesis agents as novel molecular-based therapeutics for ovarian cancer. PMID- 12079307 TI - Second-line therapy of ovarian cancer with Paclitaxel administered by both the intravenous and intraperitoneal routes: rationale and case reports. AB - BACKGROUND: A strong rationale can be proposed to support the delivery of paclitaxel by both the intravenous and the intraperitoneal routes in the management of ovarian cancer. This includes efforts to increase the concentration and duration of exposure of this cycle-specific agent within the body compartment (regional therapy) and a desire to optimize delivery of drug to tumor by capillary flow (systemic therapy). CASE REPORTS: Two patients cared for in the Gynecologic Cancer Program of the Cleveland Clinic Foundation provided an opportunity to explore, in a preliminary manner, the feasibility and toxicity of this unique approach. Both patients demonstrated reasonable tolerance of the dual route management strategy. CONCLUSION: In a carefully selected patient population, the administration of paclitaxel both systemically and regionally is a rational management strategy. Randomized controlled clinical trials will be required to determine if this approach is superior to standard intravenous drug delivery. PMID- 12079308 TI - Groin recurrence after micrometastasis in a sentinel lymph node in a patient with vulvar cancer. AB - BACKGROUND: The sentinel lymph node concept is attractive in vulvar cancer because of the potential to avoid the morbidity associated with formal groin dissection. CASE: An 84-year-old patient with a T2 carcinoma of the anterior vulva underwent surgery including bilateral sentinel node excision after identification with technetium-labeled nanocolloid. Frozen section histology showed a tumor deposit <1 mm in diameter in a left groin node whereas four nodes in the right groin were apparently negative. Completion lymphadenectomy was performed only for the left groin. Final histology including serial sectioning showed a micrometastasis in one of seven nodes from the right groin; no further treatment was given. Sixteen months postoperatively the patient developed a recurrence in the right groin; the left groin was free of tumor. CONCLUSION: This case indicates that groins with a micrometastasis detected by sentinel lymph node excision require further treatment. PMID- 12079309 TI - Adoptive immunotherapy of ovarian carcinoma. PMID- 12079310 TI - Journal of molecular biology. Introduction. PMID- 12079311 TI - A revolutionary landscape: the restructuring of biology and its convergence with medicine. PMID- 12079312 TI - Man's place in nature: evolutionary past and genomic future. PMID- 12079313 TI - The biology of the future and the future of biology. PMID- 12079314 TI - How much medical progress can we afford? Equity and the cost of health care. PMID- 12079315 TI - The life sciences and the rule of law. PMID- 12079317 TI - Science and philosophy: bridging the two cultures divide. PMID- 12079316 TI - The impact of the new biology on ethics. PMID- 12079318 TI - Stem cells, embryos and cloning--unravelling the ethics of a knotty debate. PMID- 12079319 TI - Human behavioural genetics--past and future. PMID- 12079320 TI - The Human Genome Project: a player's perspective. PMID- 12079321 TI - Acceptable intellectual property. PMID- 12079322 TI - Life sciences: discontents and consolations. PMID- 12079323 TI - Genomic metaphysics. PMID- 12079324 TI - Electronic futures in scientific communication and outreach. PMID- 12079325 TI - Is science dangerous? PMID- 12079326 TI - Scientists as communicators: how to win friends and influence people. PMID- 12079327 TI - Technology and human reproduction: 1950-2050. PMID- 12079328 TI - Still giving nature a helping hand? Surrogacy: a debate about technology and society. PMID- 12079330 TI - Production of functional single-chain Fv antibodies in the cytoplasm of Escherichia coli. AB - Production of intracellular antibodies in Escherichia coli has been thought unlikely owing to an inability to form stable disulfide bonds in the cytoplasm, a necessary step in the folding of most immunoglobulin (Ig) domains. This work investigates whether E. coli strains carrying mutations in the major intracellular disulfide bond-reduction systems (i.e. the thioredoxin and the glutathione/glutaredoxin pathways) allow the oxidation and folding of single chain variable fragment (scFv) antibodies in the cytoplasm. The effect of the co expression of disulfide bond chaperones in these cells was also examined. An scFv that recognizes the alternative sigma factor sigma(54) was used as a model to investigate disulfide bond formation and the folding of Ig domains in E. coli. The results demonstrate that functional intrabodies, with oxidized disulfide bonds in their Ig domains, are produced efficiently in E. coli cells carrying mutations in the glutathione oxidoreductase (gor) and the thioredoxin reductase (trxB) genes and co-expressing a signal-sequence-less derivative of the disulfide bond isomerase DsbC ((Delta)ssDsbC). We obtained evidence indicating that (Delta)ssDsbC acts as a chaperone promoting the correct folding and oxidation of scFvs. PMID- 12079331 TI - A novel bacteriophage-encoded RNA polymerase binding protein inhibits transcription initiation and abolishes transcription termination by host RNA polymerase. AB - Xp10 is a lytic bacteriophage of Xanthomonas oryzae, a Gram-negative bacterium that causes rice blight. We purified an Xp10 protein, p7, that binds to and inhibits X. oryzae RNA polymerase (RNAP). P7 is a novel 73 amino acid-long protein; it does not bind to and hence does not affect transcription by Escherichia coli RNAP. Analysis of E. coli/X. oryzae RNAP hybrids locates the p7 binding site to the largest X. oryzae RNAP subunit, beta'. Binding of p7 to X. oryzae RNAP holoenzyme prevents large conformational change that places the sigma subunit region 4 into the correct position for interaction with the -35 promoter element. As a result, open promoter complex formation on the -10/-35 class promoters is inhibited. Inhibition of promoter complex formation on the extended 10 class promoters is less efficient. The p7 protein also abolishes factor independent transcription termination by X. oryzae RNAP by preventing the release of nascent RNA at terminators. Further physiological and mechanistic studies of this novel transcription factor should provide additional insights into its biological role and the processes of promoter recognition and transcription termination. PMID- 12079333 TI - Structural and functional studies of an intermediate on the pathway to operator binding by Escherichia coli MetJ. AB - We present the results of in vitro DNA-binding assays for a mutant protein (Q44K) of the E. coli methionine repressor, MetJ, as well as the crystal structure at 2.2 A resolution of the apo-mutant bound to a 10-mer oligonucleotide encompassing an 8 bp met-box sequence. The wild-type protein binds natural operators co operatively with respect to protein concentration forming at least a dimer of repressor dimers along operator DNAs. The minimum operator length is thus 16 bp, each MetJ dimer interacting with a single met-box site. In contrast, the Q44K mutant protein can also bind stably as a single dimer to 8 bp target sites, in part due to additional contacts made to the phosphodiester backbone outside the 8 bp target via the K44 side-chains. Protein-protein co-operativity in the mutant is reduced relative to the wild-type allowing the properties of an intermediate on the pathway to operator site saturation to be examined for the first time. The crystal structure of the decamer complex shows a unique conformation for the protein bound to the single met-box site, possibly explaining the reduced protein protein co-operativity. In both the extended and minimal DNA complexes formed, the mutant protein makes slightly different contacts to the edges of DNA base pairs than the wild-type, even though the site of amino acid substitution is distal from the DNA-binding motif. Quantitative binding assays suggest that this is not due to artefacts caused by the crystallisation conditions but is most likely due to the relatively small contribution of such direct contacts to the overall binding energy of DNA-protein complex formation, which is dominated by sequence-dependent distortions of the DNA duplex and by the protein-protein contact between dimers. PMID- 12079332 TI - Mechanism of action of the Escherichia coli phage shock protein PspA in repression of the AAA family transcription factor PspF. AB - The PspA protein, a negative regulator of the Escherichia coli phage shock psp operon, is produced when virulence factors are exported through secretins in many Gram-negative pathogenic bacteria and its homologue in plants, VIPP1, plays a critical role in thylakoid biogenesis, essential for photosynthesis. Activation of transcription by the enhancer-dependent bacterial sigma(54) containing RNA polymerase occurs through ATP hydrolysis-driven protein conformational changes enabled by activator proteins that belong to the large AAA(+) mechanochemical protein family. We show that PspA directly and specifically acts upon and binds to the AAA(+) domain of the PspF transcription activator. Interactions involving PspF and nucleotide are changed by the action of PspA. These changes and the complexes that form between PspF and PspA can explain how PspA exerts its negative effects upon transcription activated by PspF, and are of significance when considering how activities of other AAA(+) proteins might be controlled. PMID- 12079334 TI - Structure of a beta-alanine-linked polyamide bound to a full helical turn of purine tract DNA in the 1:1 motif. AB - Polyamides composed of N-methylpyrrole (Py), N-methylimidazole (Im) and N methylhydroxypyrrole (Hp) amino acids linked by beta-alanine (beta) bind the minor groove of DNA in 1:1 and 2:1 ligand to DNA stoichiometries. Although the energetics and structure of the 2:1 complex has been explored extensively, there is remarkably less understood about 1:1 recognition beyond the initial studies on netropsin and distamycin. We present here the 1:1 solution structure of ImPy-beta Im-beta-ImPy-beta-Dp bound in a single orientation to its match site within the DNA duplex 5'-CCAAAGAGAAGCG-3'.5'-CGCTTCTCTTTGG-3' (match site in bold), as determined by 2D (1)H NMR methods. The representative ensemble of 12 conformers has no distance constraint violations greater than 0.13 A and a pairwise RMSD over the binding site of 0.80 A. Intermolecular NOEs place the polyamide deep inside the minor groove, and oriented N-C with the 3'-5' direction of the purine rich strand. Analysis of the high-resolution structure reveals the ligand bound 1:1 completely within the minor groove for a full turn of the DNA helix. The DNA is B-form (average rise=3.3 A, twist=38 degrees ) with a narrow minor groove closing down to 3.0-4.5 A in the binding site. The ligand and DNA are aligned in register, with each polyamide NH group forming bifurcated hydrogen bonds of similar length to purine N3 and pyrimidine O2 atoms on the floor of the minor groove. Each imidazole group is hydrogen bonded via its N3 atom to its proximal guanine's exocyclic amino group. The important roles of beta-alanine and imidazole for 1:1 binding are discussed. PMID- 12079335 TI - The isomerization of the UvrB-DNA preincision complex couples the UvrB and UvrC activities. AB - In Escherichia coli nucleotide excision repair, the UvrB-DNA preincision complex plays a key role, linking adduct recognition to incision. We previously showed that the efficiency of the incision is inversely related to the stability of the preincision complex. We postulated that an isomerization reaction converts [UvrB DNA], stable but incompetent for incision, into the [UvrB-DNA]' complex, unstable and competent for incision. Here, we identify two parameters, negative supercoiling and presence of a nick at the fifth phosphodiester bond 3' to the lesion, that accelerate the isomerization leading to an increasing incision efficiency. We also show that the [UvrB-DNA] complex is more resistant to a salt concentration increase than the [UvrB-DNA]' complex. Finally, we report that the [UvrB-DNA]' is recognized by UvrC. These data suggest that the isomerization reaction leads to an exposure of single-stranded DNA around the lesion. This newly exposed single-stranded DNA serves as a binding site and substrate for the UvrC endonuclease. We propose that the isomerization reaction is responsible for coupling UvrB and UvrC activities and that this reaction corresponds to the binding of ATP. PMID- 12079336 TI - Evolution of an antibiotic resistance enzyme constrained by stability and activity trade-offs. AB - Pressured by antibiotic use, resistance enzymes have been evolving new activities. Does such evolution have a cost? To investigate this question at the molecular level, clinically isolated mutants of the beta-lactamase TEM-1 were studied. When purified, mutant enzymes had increased activity against cephalosporin antibiotics but lost both thermodynamic stability and kinetic activity against their ancestral targets, penicillins. The X-ray crystallographic structures of three mutant enzymes were determined. These structures suggest that activity gain and stability loss is related to an enlarged active site cavity in the mutant enzymes. In several clinically isolated mutant enzymes, a secondary substitution is observed far from the active site (Met182-->Thr). This substitution had little effect on enzyme activity but restored stability lost by substitutions near the active site. This regained stability conferred an advantage in vivo. This pattern of stability loss and restoration may be common in the evolution of new enzyme activity. PMID- 12079337 TI - The projection domain of MAP4 suppresses the microtubule-bundling activity of the microtubule-binding domain. AB - Microtubule-associated protein 4 (MAP4), a major MAP expressed in proliferating non-neuronal cells, consists of an N-terminal projection (PJ) domain and a C terminal microtubule-binding (MTB) domain. The PJ domain of MAP4 is divided into three regions; the N-terminal acidic region (the Na-region), the multiple KDM repeated sequence region (the KDM-region), and the b-region followed by the MTB domain. To investigate roles of the PJ domain, we prepared three truncated forms of human MAP4 with different PJ domain lengths; PJ1, PJ2 and MTB with deletion of about one-third, two-third and all of the PJ domain, respectively, and examined their effects on bundle formation of microtubules (MTs). MTs polymerized by full length MAP4 were singly distributed as observed by both negative staining electron microscopy and dark field microscopy. MTs with PJ1 were also separated in solution but became pairs when pelleted by centrifugation. PJ2 formed planar two-dimensional bundles consisting of several MTs (the 2D-bundle). MTB induced large bundles of many MTs, tightly packed without space in between (termed the 3D bundle). To study how the PJ domain decreases the bundle-forming activity of the MTB domain of MAP4, we made three additional deletion-mutants of MAP4, called Na MTB, KDM-MTB and Na-PJ2. Na-MTB and KDM-MTB, in which the KDM/b-region and both of Na- and b-regions were deleted respectively, were prepared by fusing the Na region or KDM-region to MTB. Both of Na-MTB and KDM-MTB suppressed the 3D-bundle formation as effectively as PJ2. MTs polymerized with Na-PJ2, the KDM-deletion mutant made by adding the Na-region to PJ2, were singular and did not become bundles. These results indicated that the PJ domain kept individual MTs separated by suppressing the bundle-forming ability of the MTB domain. The suppressive activity of the PJ domain was correlated with the length, but not the amino acid sequence, of the PJ. PMID- 12079338 TI - Reversible dimer formation and stability of the anti-tumour single-chain Fv antibody MFE-23 by neutron scattering, analytical ultracentrifugation, and NMR and FT-IR spectroscopy. AB - MFE-23 is a single chain Fv (scFv) antibody molecule used to target colorectal cancer through its high affinity for the tumour marker carcinoembryonic antigen (CEA). ScFv molecules are formed from peptide-linked antibody V(H) and V(L) domains, and many of these form dimers. Our recent crystal structure for MFE-23 showed that this formed an unusual symmetric back-to-back association of two monomers that is consistent with a domain-swapped diabody structure. Neutron scattering and modelling fits showed that MFE-23 existed as compact V(H)-V(L) linked monomers at therapeutically relevant concentrations below 1 mg/ml. Size exclusion gel chromatography showed that the monomeric and dimeric forms of MFE 23 could be separated, and that the proportions of these two forms depended on the starting MFE-23 concentration. Sedimentation equilibrium experiments by analytical ultracentrifugation at nine concentrations of MFE-23 indicated a reversible monomer-dimer self-association equilibrium with an association constant of 1.9x10(3)-2.2x10(3) M(-1). Sedimentation velocity experiments using the time derivative g(s(*)) method showed that MFE-23-His has a concentration dependent weight average sedimentation coefficient that increased from 1.8 S for the monomer to about 3-6 S for the dimer. Both values agreed with those calculated from the MFE-23 crystal structure. In relation to the thermal stability of MFE-23, denaturation experiments by (1)H NMR and FT-IR spectroscopy showed that the molecule is stable up to 47 degrees C, after which denaturation was irreversible. MFE-23 dimerisation is discussed in terms of a new model for diabody structures, in which the V(H) and V(L) domains in the monomer are able to dissociate and reassociate to form a dimer, or diabody, but in which symmetric back-to-back contacts between the two monomers are formed. This dimerisation in solution is attributed to the complementary nature of the C-terminal surface of the MFE-23 monomer. Crystal structures for seven other scFv molecules have shown that, while the contact residues for symmetric back-to-back dimer formation in MFE-23 are not fully conserved, in principle, back-to-back contacts can be formed in these too. This offers possibilities for the creation of other forms of scFv molecules. PMID- 12079339 TI - Archaeal Sm proteins form heptameric and hexameric complexes: crystal structures of the Sm1 and Sm2 proteins from the hyperthermophile Archaeoglobus fulgidus. AB - Proteins of largely unknown function related to the Sm proteins present in the core domain of eukaryotic small nuclear ribonucleoprotein particles have recently been detected in Archaea. In contrast to eukaryotes, Archaea contain maximally two distinct Sm-related proteins belonging to different subfamilies, we refer to as Sm1 and Sm2. Here we report the crystal structures of the Sm1- and Sm2-type proteins from the hyperthermophilic euryarchaeon Archaeoglobus fulgidus (AF-Sm1 and AF-Sm2) at a resolution of 2.5 and 1.95 A, respectively. While the AF-Sm1 protein forms a heptameric ring structure similar to that found in other archaeal Sm1-type proteins, the AF-Sm2 protein unexpectedly forms a homo-hexamer in the crystals, and, as is evident from the mass spectrometric analysis, also in solution. Both proteins have essentially the same monomer fold and inter-subunit beta-sheet hydrogen bonding giving rise to a similar overall architecture of the doughnut-shaped six and seven-membered rings. In addition, a conserved uracil binding pocket identified previously in an AF-Sm1/RNA complex, suggests a common RNA-binding mode for the AF-Sm1 and AF-Sm2 proteins, in line with solution studies showing preferential binding to U-rich oligonucleotides for both proteins. Clear differences are however seen in the charge distribution within the two structures. The rough faces of the rings, i.e. the faces not containing the base binding pockets, have opposite charges in the two structures, being predominantly positive in AF-Sm1 and negative in AF-Sm2. Differences in the ionic interactions between subunits provide an explanation for the distinctly different oligomerisation behaviour of the AF-Sm1 and AF-Sm2 proteins and of Sm1- and Sm2 type proteins in general, as well as the stability of their complexes. Implications for the functions of archaeal Sm proteins are being discussed. PMID- 12079340 TI - X-ray structure of a dihydropyrimidinase from Thermus sp. at 1.3 A resolution. AB - Dihydropyrimidinases (hydantoinases) catalyse the reversible hydrolytic ring opening of cyclic diamides such as dihydropyrimidines in the catabolism of pyrimidines. In biotechnology, these enzymes find application in the enantiospecific production of amino acids from racemic hydantoins. The crystal structure of a D-enantio-specific dihydropyrimidinase from Thermus sp. (D hydantoinase) was solved de novo by multiwavelength anomalous diffraction phasing. In spite of a large unit cell the D-hydantoinase crystals exhibit excellent diffraction properties. The structure was subsequently refined at 1.30 A resolution against native data. The core of D-hydantoinase consists of a (alpha/beta)(8)-barrel, which is flanked by a beta-sheet domain and some additional helices. In the active site, a carboxylated lysine residue and the catalytically active hydroxide ion bridge a binuclear zinc centre. The tertiary structure and shape of the active site show strong homology to that of ureases, dihydroorotases, and phosphotriesterases. The homology of the active site was exploited for in silicio docking of substrates in the active site. This could shed light both on the substrate binding in hydantoinases and on the recently highly discussed origin of the proton in the course of hydantoinase catalysis. PMID- 12079342 TI - Localized unfolding of collagen explains collagenase cleavage near imino-poor sites. AB - Collagenases cleave all three chains of type III collagen at specific sites characterized by a Gly-Leu or a Gly-Ile bond that is upstream from an imino acid poor region. Molecular dynamics trajectories were used to calculate the free energy of unfolding for collagen-like model peptides. The free energy profiles suggest that such imino-poor regions can adopt a low-energy, partially unfolded state where one of the peptide chains forms a solvent-exposed loop. The results are consistent with a model for collagenase cleavage where partial unfolding of imino-poor regions enables collagenases to gain access to their cleavage sites. PMID- 12079343 TI - In vitro transcription and start site selection in Schizosaccharomyces pombe. AB - We have used the fission yeast Schizosaccharomyces pombe to establish both a biochemical and genetic system to study the roles of general transcription factors in transcription initiation. Extracts were prepared that faithfully transcribed S. pombe promoters and the results confirm that, in contrast to the budding yeast Saccharomyces cerevisiae, in vitro transcription in S. pombe initiates near to the TATA element. S. pombe transcription relies on upstream activation sequence elements and these can be replaced successfully with sites for binding Gal4-VP16 activators. Although it is mammalian-like in these respects, S. pombe initiation uses an unusual scanning mechanism. This directs initiation, preferentially using purines, within a narrow window approximately 25 40 base-pairs downstream from the edge of the TATA element. Genetic experiments showed that this scanning mechanism was associated with the properties of the TFIIB polypeptide. When human TFIIB was expressed in S. pombe, it was accepted by the endogenous transcription machinery and caused initiation to be restricted to the closer edge of this window, corresponding to the distance in humans. Preliminary experiments suggested that S. cerevisiae TFIIB was not accepted. The results enlarge the potential for using fission yeast to study the properties of general transcription factors such as TFIIB in choosing the sites at which transcription initiates. PMID- 12079344 TI - Solution conformations of unmodified and A(37)N(6)-dimethylallyl modified anticodon stem-loops of Escherichia coli tRNA(Phe). AB - The modification of RNA nucleotide bases, a fundamental process in all cells, alters the chemical and physical properties of RNA molecules and broadly impacts the physiological properties of cells. tRNA molecules are by far the most diverse modified RNA species within cells, containing as a group >80% of the known 96 chemically unique nucleic acid modifications. The greatest varieties of modifications are located on residue 37 and play a role in ensuring fidelity and efficiency of protein synthesis. The enzyme dimethylallyl (Delta(2)-isopentenyl) diphosphate:tRNA transferase catalyzes the addition of a dimethylallyl group to the exocyclic amine nitrogen (N6) of A(37) in several tRNA species. Using a 17 residue oligoribonucleotide corresponding to the anticodon arm of Escherichia coli tRNA(Phe), we have investigated the structural and dynamic changes introduced by the dimethylallyl group. The unmodified RNA molecule adopts stem loop conformation composed of seven base-pairs and a compact three nucleotide loop. This conformation is distinctly different from the U-turn motif that characterizes the anticodon arm in the X-ray crystal structure of the fully modified yeast tRNA(Phe). The adoption of the tri-nucleotide loop by the purine rich unmodified tRNA(Phe) anticodon arm suggests that other anticodon sequences, especially those containing pyrimidine bases, also may favor a tri-loop conformation. Introduction of the dimethylallyl modification increases the mobility of nucleotides of the loop region but does not dramatically alter the RNA conformation. The dimethylallyl modification may enhance ribosome binding through multiple mechanisms including destabilization of the closed anticodon loop and stabilization of the codon-anticodon helix. PMID- 12079345 TI - RNA recognition by transcriptional antiterminators of the BglG/SacY family: mapping of SacY RNA binding site. AB - Transcriptional antiterminators of the BglG/SacY family are bacterial regulatory proteins able to prevent the premature arrest of transcription through specific binding to a ribonucleic antiterminator (RAT) sequence. The RNA recognition module of these regulators is made of the 55-amino acid long N-terminal domain which can by itself promote efficient antitermination activity in vivo and RNA binding in vitro. The structure of this domain, which was called CAT for co antiterminator, has first been determined for SacY from Bacillus subtilis and the putative surface contacting RNA has been defined by NMR footprinting. Here we have performed a genetic mapping of the SacY-CAT RNA binding site by substituting 24 amino acid residues including those previously identified by NMR, the highly conserved residues in the 55 homologous antiterminators recognised in the databases and all the positively charged residues. A total of 57 SacY-CAT variants have been constructed and tested in vivo for their antitermination efficiency. A few of these variants were then purified in order to analyse their RNA binding properties by surface plasmon resonance and to check their structural integrity by NMR. The present study validates and clarifies the RNA interacting surface previously mapped by NMR. The residues that are the most intolerant to substitutions, Asn8, His9, Asn10, Gly25, Gly27, and Phe30, are aligned across the CAT dimer interface and form the core of the RNA binding site. Three highly conserved residues stand outside the interaction surface but are essential for maintaining the CAT dimeric structure (Phe47) or may play an important functional role in the full length protein (Glu20 and Lys32). Interestingly, none of the twelve positively charged residues of SacY-CAT are crucial for the antitermination activity. By replacing three Lys residues and combining the Ala26 ->Arg mutation that significantly enhanced the affinity for RNA, we engineered a SacY-CAT variant that should be suitable for NMR study of the complex. PMID- 12079346 TI - Crystal structure of TB-RBP, a novel RNA-binding and regulating protein. AB - The testis/brain-RNA-binding protein (TB-RBP) spatially and temporally controls the expression of specific mRNAs in developing male germ cells and brain cells, and is implicated in DNA recombination and repair events. We report the 2.65 A crystal structure of mouse TB-RBP. The structure is predominantly alpha-helical and exhibits a novel protein fold and mode of assembly. Crystal symmetry and molecular symmetry combine to form an octet of TB-RBP monomers in the shape of an elongated spherical particle with a large cavity at its center. Amino acid residues that affect RNA and DNA binding are located on the interior surface of the assembled particle, and a putative nucleotide-binding domain that controls RNA binding is located at a dimer interface. Other modes of assembly are suggested for TB-RBP based on our structure and recently reported electron microscopic reconstructions of human TB-RBP. PMID- 12079347 TI - Secondary structure prediction for aligned RNA sequences. AB - Most functional RNA molecules have characteristic secondary structures that are highly conserved in evolution. Here we present a method for computing the consensus structure of a set aligned RNA sequences taking into account both thermodynamic stability and sequence covariation. Comparison with phylogenetic structures of rRNAs shows that a reliability of prediction of more than 80% is achieved for only five related sequences. As an application we show that the Early Noduline mRNA contains significant secondary structure that is supported by sequence covariation. PMID- 12079348 TI - Beta subunit residues 186-433 and 436-445 are commonly used by Esigma54 and Esigma70 RNA polymerase for open promoter complex formation. AB - During transcription initiation by DNA-dependent RNA polymerase (RNAP) promoter DNA has to be melted locally to allow the synthesis of RNA transcript. Localized melting of promoter DNA is a target for genetic regulation and is poorly understood at the molecular level. The Escherichia coli RNAP holoenzyme is a six subunit (alpha(2)betabeta'omegasigma; Esigma) protein complex. The sigma subunit is directly responsible for promoter recognition and contributes to localized DNA melting. Mutations in the beta subunit have profound effects on promoter melting by Esigma70. The sigma54 subunit is a representative of an unrelated class of the sigma subunits. Here, we determined whether mutations in the beta subunit that affect late stages of promoter complex formation by Esigma70 also influence promoter complex formation by the enhancer-dependent Esigma54. Analyses of in vitro defects in promoter complex formation and transcription initiation exhibited by mutant Esigma54 suggest that during promoter complex formation by Esigma54 and Esigma70 a common set of beta subunit sequences is used. Late stages of promoter complex formation and localized melting of promoter DNA by Esigma70 and Esigma54 thus proceed through a common pathway. PMID- 12079349 TI - The Escherichia coli dam DNA methyltransferase modifies DNA in a highly processive reaction. AB - The Escherichia coli dam adenine-N6 methyltransferase modifies DNA at GATC sequences. It is involved in post-replicative mismatch repair, control of DNA replication and gene regulation. We show that E. coli dam acts as a functional monomer and methylates only one strand of the DNA in each binding event. The preferred way of ternary complex assembly is that the enzyme first binds to DNA and then to S-adenosylmethionine. The enzyme methylates an oligonucleotide containing two dam sites and a 879 bp PCR product with four sites in a fully processive reaction. On lambda-DNA comprising 48,502 bp and 116 dam sites, E. coli dam scans 3000 dam sites per binding event in a random walk, that on average leads to a processive methylation of 55 sites. Processive methylation of DNA considerably accelerates DNA methylation. The highly processive mechanism of E. coli dam could explain why small amounts of E. coli dam are able to maintain the methylation state of dam sites during DNA replication. Furthermore, our data support the general rule that solitary DNA methyltransferase modify DNA processively whereas methyltransferases belonging to a restriction-modification system show a distributive mechanism, because processive methylation of DNA would interfere with the biological function of restriction-modification systems. PMID- 12079351 TI - Complete nucleotide sequence and likely recombinatorial origin of bacteriophage T3. AB - We report the complete genome sequence (38,208 bp) of bacteriophage T3 and provide a bioinformatic comparative analysis with other completely sequenced members of the T7 group of phages. This comparison suggests that T3 has evolved from a recombinant between a T7-like coliphage and a yersiniophage. To assess this, recombination between T7 and the Yersinia enterocolitica serotype O:3 phage phiYeO3-12 was accomplished in vivo; coliphage progeny from this cross were selected that had many biological properties of T3. This represents the first experimentally observed recombination between lytic phages whose normal hosts are different bacterial genera. PMID- 12079350 TI - Solvent mediated interactions in the structure of the nucleosome core particle at 1.9 a resolution. AB - Solvent binding in the nucleosome core particle containing a 147 base pair, defined-sequence DNA is characterized from the X-ray crystal structure at 1.9 A resolution. A single-base-pair increase in DNA length over that used previously results in substantially improved clarity of the electron density and accuracy for the histone protein and DNA atomic coordinates. The reduced disorder has allowed for the first time extensive modeling of water molecules and ions. Over 3000 water molecules and 18 ions have been identified. Water molecules acting as hydrogen-bond bridges between protein and DNA are approximately equal in number to the direct hydrogen bonds between these components. Bridging water molecules have a dual role in promoting histone-DNA association not only by providing further stability to direct protein-DNA interactions, but also by enabling formation of many additional interactions between more distantly related elements. Water molecules residing in the minor groove play an important role in facilitating insertion of arginine side-chains. Water structure at the interface of the histones and DNA provides a means of accommodating intrinsic DNA conformational variation, thus limiting the sequence dependency of nucleosome positioning while enhancing mobility. Monovalent anions are bound near the N termini of histone alpha-helices that are not occluded by DNA phosphate groups. Their location in proximity to the DNA phosphodiester backbone suggests that they damp the electrostatic interaction between the histone proteins and the DNA. Divalent cations are bound at specific sites in the nucleosome core particle and contribute to histone-histone and histone-DNA interparticle interactions. These interactions may be relevant to nucleosome association in arrays. PMID- 12079352 TI - Mechanism of coenzyme recognition and binding revealed by crystal structure analysis of ferredoxin-NADP+ reductase complexed with NADP+. AB - The flavoenzyme ferredoxin-NADP+ reductase (FNR) catalyses the production of NADPH in photosynthesis. The three-dimensional structure of FNR presents two distinct domains, one for binding of the FAD prosthetic group and the other for NADP+ binding. In spite of extensive experiments and different crystallographic approaches, many aspects about how the NADP+ substrate binds to FNR and how the hydride ion is transferred from FAD to NADP+ remain unclear. The structure of an FNR:NADP+ complex from Anabaena has been determined by X-ray diffraction analysis of the cocrystallised units to 2.1 A resolution. Structural perturbation of FNR induced by complex formation produces a narrower cavity in which the 2'-phospho AMP and pyrophosphate portions of the NADP+ are perfectly bound. In addition, the nicotinamide mononucleotide moiety is placed in a new pocket created near the FAD cofactor with the ribose being in a tight conformation. The crystal structure of this FNR:NADP+ complex obtained by cocrystallisation displays NADP+ in an unusual conformation and can be considered as an intermediate state in the process of coenzyme recognition and binding. Structural analysis and comparison with previously reported complexes allow us to postulate a mechanism which would permit efficient hydride transfer to occur. Besides, this structure gives new insights into the postulated formation of the ferredoxin:FNR:NADP+ ternary complex by prediction of new intermolecular interactions, which could only exist after FNR:NADP+ complex formation. Finally, structural comparison with the members of the broad FNR structural family also provides an explanation for the high specificity exhibited by FNR for NADP+/H versus NAD+/H. PMID- 12079353 TI - Differential oligosaccharide recognition by evolutionarily-related beta-1,4 and beta-1,3 glucan-binding modules. AB - Enzymes active on complex carbohydrate polymers frequently have modular structures in which a catalytic domain is appended to one or more carbohydrate binding modules (CBMs). Although CBMs have been classified into a number of families based upon sequence, many closely related CBMs are specific for different polysaccharides. In order to provide a structural rationale for the recognition of different polysaccharides by CBMs displaying a conserved fold, we have studied the thermodynamics of binding and three-dimensional structures of the related family 4 CBMs from Cellulomonas fimi Cel9B and Thermotoga maritima Lam16A in complex with their ligands, beta-1,4 and beta-1,3 linked gluco oligosaccharides, respectively. These two CBMs use a structurally conserved constellation of aromatic and polar amino acid side-chains that interact with sugars in two of the five binding subsites. Differences in the length and conformation of loops in non-conserved regions create binding-site topographies that complement the known solution conformations of their respective ligands. Thermodynamics interpreted in the light of structural information highlights the differential role of water in the interaction of these CBMs with their respective oligosaccharide ligands. PMID- 12079354 TI - Muscle Z-band ultrastructure: titin Z-repeats and Z-band periodicities do not match. AB - Vertebrate muscle Z-bands show zig-zag densities due to different sets of alpha actinin cross-links between anti-parallel actin molecules. Their axial extent varies with muscle and fibre type: approximately 50 nm in fast and approximately 100 nm in cardiac and slow muscles, corresponding to the number of alpha-actinin cross-links present. Fish white (fast) muscle Z-bands have two sets of alpha actinin links, mammalian slow muscle Z-bands have six. The modular structure of the approximately 3 MDa protein titin that spans from M-band to Z-band correlates with the axial structure of the sarcomere; it may form the template for myofibril assembly. The Z-band-located amino-terminal 80 kDa of titin includes 45 residue repeating modules (Z-repeats) that are expressed differentially; heart, slow and fast muscles have seven, four to six and two to four Z-repeats, respectively. Gautel et al. proposed a Z-band model in which each Z-repeat links to one level of alpha-actinin cross-links, requiring that the axial extent of a Z-repeat is the same as the axial separation of alpha-actinin layers, of which there are two in every actin crossover repeat. The span of a Z-repeat in vitro is estimated by Atkinson et al. to be 12 nm or less; much less than half the normal vertebrate muscle actin crossover length of 36 nm. Different actin-binding proteins can change this length; it is reduced markedly by cofilin binding, or can increase to 38.5 nm in the abnormally large nemaline myopathy Z-band. Here, we tested whether in normal vertebrate Z-bands there is a marked reduction in crossover repeat so that it matches twice the apparent Z-repeat length of 12 nm. We found that the measured periodicities in wide Z-bands in slow and cardiac muscles are all very similar, about 39 nm, just like the nemaline myopathy Z-bands. Hence, the 39 nm periodicity is an important conserved feature of Z-bands and either cannot be explained by titin Z-repeats as previously suggested or may correlate with two Z repeats. PMID- 12079355 TI - Activation of the nicotinic acetylcholine receptor involves a switch in conformation of the alpha subunits. AB - The nicotinic acetylcholine (ACh) receptor belongs to a superfamily of synaptic ion channels that open in response to the binding of chemical transmitters. Their mechanism of activation is not known in detail, but a time-resolved electron microscopic study of the muscle-type ACh receptor had suggested that a local disturbance in the ligand-binding region and consequent rotations in the ligand binding alpha subunits, connecting to the transmembrane portion, are involved. A more precise interpretation of this structural change is given here, based on comparison of the extracellular domain of the ACh receptor with an ACh-binding protein (AChBP) to which a putative agonist is bound. We find that, to a good approximation, there are two alternative extended conformations of the ACh receptor subunits, one characteristic of either alpha subunit before activation, and the other characteristic of all three non-alpha subunits and the protomer of AChBP. Substitution in the three-dimensional maps of alpha by non-alpha subunits mimics the changes seen on activation, suggesting that the structures of the alpha subunits are modified initially by their interactions with neighbouring subunits and switch to the non-alpha form when ACh binds. This structural change, which entails 15-16 degrees rotations of the inner pore-facing parts of the alpha subunits, most likely acts as the trigger that opens the gate in the membrane spanning pore. PMID- 12079356 TI - Ions and counterions in a biological channel: a molecular dynamics simulation of OmpF porin from Escherichia coli in an explicit membrane with 1 M KCl aqueous salt solution. AB - A 5 ns all-atom molecular dynamics trajectory of Escherichia coli OmpF porin embedded in an explicit dimyristoyl-phosphatidylcholine (DMPC) bilayer bathed by a 1 M [KCl] aqueous salt solution is generated to explore the microscopic details of the mechanism of ion permeation. The atomic model includes the OmpF trimer, 124 DMPC, 13470 water molecules as well as 231 K+ and 201 Cl-, for a total of 70,693 atoms. The structural and dynamical results are in excellent agreement with the X-ray data. The global root-mean-square deviation of the backbone atoms relative to the X-ray structure is 1.4 A. A cluster of three fully charged arginine (Arg42, Arg82, and Arg132) facing two acidic residues (Asp113 and Glu117) on L3 in the narrowest part of the aqueous pore is observed to be very stable in the crystallographic conformation. In this region of the pore, the water molecules are markedly oriented perpendicular to the channel axis due to the strong transversal electrostatic field arising from those residues. On average the size of the pore is smaller during the simulation than in the X-ray structure, undergoing small fluctuations. No large movements of loop L3 leading to a gating of the pore are observed. Remarkably, it is observed that K+ and Cl- follow two well-separated average pathways spanning over nearly 40 A along the axis of the pore. In the center of the monomer, the two screw-like pathways have a left-handed twist, undergoing a counter-clockwise rotation of 180 degrees from the extracellular vestibule to the pore periplasmic side. In the pore, the dynamical diffusion constants of the ions are reduced by about 50% relative to their value in bulk solvent. Analysis of ion solvation across the channel reveals that the contributions from the water and the protein are complementary, keeping the total solvation number of both ions nearly constant. Unsurprisingly, K+ have a higher propensity to occupy the aqueous pore than Cl-, consistent with the cation selectivity of the channel. However, further analysis suggests that ion ion pairs play an important role. In particular, it is observed that the passage of Cl- occurs only in the presence of K+ counterions, and isolated K+ can move through the channel and permeate on their own. The presence of K+ in the pore screens the negative electrostatic potential arising from OmpF to help the translocation of Cl- by formation of ion pairs. PMID- 12079358 TI - Impact of mutations affecting ND mitochondria-encoded subunits on the activity and assembly of complex I in Chlamydomonas. Implication for the structural organization of the enzyme. AB - The mitochondrial rotenone-sensitive NADH:ubiquinone oxidoreductase (complex I) comprises more than 35 subunits, the majority of which are encoded by the nucleus. In Chlamydomonas reinhardtii, only five components (ND1, ND2, ND4, ND5 and ND6) are coded for by the mitochondrial genome. Here, we characterize two mitochondrial mutants (dum5 and dum17) showing strong reduction or inactivation of complex I activity: dum5 is a 1T deletion in the 3' UTR of nd5 whereas dum17 is a 1T deletion in the coding sequence of nd6. The impact of these mutations and of mutations affecting nd1, nd4 and nd4/nd5 genes on the assembly of complex I is investigated. After separation of the respiratory complexes by blue native (BN) PAGE or sucrose gradient centrifugation, we demonstrate that the absence of intact ND1 or ND6 subunit prevents the assembly of the 850 kDa whole complex, whereas the loss of ND4 or ND4/ND5 leads to the formation of a subcomplex of 650 kDa present in reduced amount. The implications of our findings for the possible role of these ND subunits on the activity of complex I and for the structural organization of the membrane arm of the enzyme are discussed. In mitochondria from all the strains analyzed, we moreover detected a 160-210 kDa fragment comprising the hydrophilic 49 kDa and 76 kDa subunits of the complex I peripheral arm and showing NADH dehydrogenase activity. PMID- 12079357 TI - Crystal structure of a biliverdin IXalpha reductase enzyme-cofactor complex. AB - Biliverdin reductase (BVR) catalyzes the last step in heme degradation by reducing the gamma-methene bridge of the open tetrapyrrole, biliverdin IXalpha, to bilirubin with the concomitant oxidation of a beta-nicotinamide adenine dinucleotide (NADH) or beta-nicotinamide adenine dinucleotide phosphate (NADPH) cofactor. Bilirubin is the major bile pigment in mammals and has antioxidant and anticompliment activity. We have determined X-ray crystal structures of apo rat BVR and its complex with NADH at 1.2 A and 1.5 A resolution, respectively. In agreement with an independent structure determination of the apo-enzyme, BVR consists of an N-terminal dinucleotide-binding domain (Rossmann-fold) and a C terminal domain that contains a six-stranded beta-sheet that is flanked on one face by several alpha-helices. The C-terminal and N-terminal domains interact extensively, forming the active site cleft at their interface. The cofactor complex structure reported here reveals that the cofactor nicotinamide ring extends into the active site cleft, where it is adjacent to conserved amino acid residues and, consistent with the known stereochemistry of the reaction catalyzed by BVR, the si face of the ring is accessible for hydride transfer. The only titratable side-chain that appears to be suitably positioned to function as a general acid in catalysis is Tyr97. This residue, however, is not essential for catalysis, since the Tyr97Phe mutant protein retains 50% activity. This finding suggests that the dominant role in catalysis may be performed by hydride transfer from the cofactor, a process that may be promoted by proximity of the invariant residues Glu96, Glu123, and Glu126, to the nicotinamide ring. PMID- 12079359 TI - Crystal structures of two homologous pathogenesis-related proteins from yellow lupine. AB - Pathogenesis-related class 10 (PR10) proteins are restricted to the plant kingdom where they are coded by multigene families and occur at high levels. In spite of their abundance, their physiological role is obscure although members of a distantly related subclass (cytokinin-specific binding proteins) are known to bind plant hormones. PR10 proteins are of special significance in legume plants where their expression patterns are related to infection by the symbiotic, nitrogen-fixing bacteria. Here we present the first crystal structures of classic PR10 proteins representing two homologues from one subclass in yellow lupine. The general fold is similar and, as in a birch pollen allergen, consists of a seven stranded beta-sheet wrapped around a long C-terminal helix. The mouth of a large pocket formed between the beta-sheet and the helix seems a likely site for ligand binding. The shape of the pocket varies because, in variance with the rigid beta sheet, the helix shows unusual conformational variability consisting in bending, disorder, and axial shifting. A surface loop, proximal to the entrance to the internal cavity, shows an unusual structural conservation and rigidity in contrast to the high glycine content in its sequence. The loop is different from the so-called glycine-rich P-loops that bind phosphate groups of nucleotides, but it is very likely that it does play a role in ligand binding in PR10 proteins. PMID- 12079360 TI - Structural properties of a collagenous heterotrimer that mimics the collagenase cleavage site of collagen type I. AB - Collagens contain sequence- and conformation-dependent epitopes responsible for their digestion by collagenases at specific loci. A synthetic heterotrimer construct containing the collagenase cleavage site of collagen type I was found to mimic perfectly native collagen in terms of selectivity and mode of enzymatic degradation. The NMR conformational analysis of this molecule clearly revealed the presence of two structural domains, i.e. a triple helix spanning the Gly-Pro Hyp repeats and a less ordered portion corresponding to the collagenase cleavage site where the three chains are aligned in extended conformation with loose interchain contacts. These structural properties allow for additional insights into the very particular mechanism of collagen digestion by collagenases. PMID- 12079361 TI - Solution structures of UBA domains reveal a conserved hydrophobic surface for protein-protein interactions. AB - UBA domains are a commonly occurring sequence motif of approximately 45 amino acid residues that are found in diverse proteins involved in the ubiquitin/proteasome pathway, DNA excision-repair, and cell signaling via protein kinases. The human homologue of yeast Rad23A (HHR23A) is one example of a nucleotide excision-repair protein that contains both an internal and a C terminal UBA domain. The solution structure of HHR23A UBA(2) showed that the domain forms a compact three-helix bundle. We report the structure of the internal UBA(1) domain of HHR23A. Comparison of the structures of UBA(1) and UBA(2) reveals that both form very similar folds and have a conserved large hydrophobic surface patch. The structural similarity between UBA(1) and UBA(2), in spite of their low level of sequence conservation, leads us to conclude that the structural variability of UBA domains in general is likely to be rather small. On the basis of the structural similarities as well as analysis of sequence conservation, we predict that this hydrophobic surface patch is a common protein-interacting surface present in diverse UBA domains. Furthermore, accumulating evidence that ubiquitin binds to UBA domains leads us to the prediction that the hydrophobic surface patch of UBA domains interacts with the hydrophobic surface on the five-stranded beta-sheet of ubiquitin. Detailed comparison of the structures of the two UBA domains, combined with previous mutagenesis studies, indicates that the binding site of HIV-1 Vpr on UBA(2) does not completely overlap the ubiquitin binding site. PMID- 12079362 TI - Prediction of human protein function from post-translational modifications and localization features. AB - We have developed an entirely sequence-based method that identifies and integrates relevant features that can be used to assign proteins of unknown function to functional classes, and enzyme categories for enzymes. We show that strategies for the elucidation of protein function may benefit from a number of functional attributes that are more directly related to the linear sequence of amino acids, and hence easier to predict, than protein structure. These attributes include features associated with post-translational modifications and protein sorting, but also much simpler aspects such as the length, isoelectric point and composition of the polypeptide chain. PMID- 12079363 TI - Folding and oxidation of the antibody domain C(H)3. AB - The non-covalent homodimer formed by the C-terminal domains of the IgG1 heavy chains (C(H)3) is the simplest naturally occurring model system for studying immunoglobulin folding and assembly. In the native state, the intrachain disulfide bridge, which connects a three-stranded and a four-stranded beta-sheet is buried in the hydrophobic core of the protein. Here, we show that the disulfide bridge is not required for folding and association, since the reduced C(H)3 domain folds to a dimer with defined secondary and tertiary structure. However, the thermodynamic stability of the reduced C(H)3 dimer is much lower than that of the oxidized state. This allows the formation of disulfide bonds either concomitant with folding (starting from the reduced, denatured state) or after folding (starting from the reduced dimer). The analysis of the two processes revealed that, under all conditions investigated, one of the cysteine residues, Cys 86, reacts preferentially with oxidized glutathione to a mixed disulfide that subsequently interacts with the less-reactive second thiol group of the intra-molecular disulfide bond. For folded C(H)3, the second step in the oxidation process is slow. In contrast, starting from the unfolded and reduced protein, the oxidation reaction is faster. However, the overall folding reaction of C(H)3 during oxidative folding is a slow process. Especially, dimerization is slow, compared to the association starting from the denatured oxidized state. This deceleration may be due to misfolded conformations trapped by the disulfide bridge. PMID- 12079366 TI - Recurrent habitat disturbance and species diversity in a multiple-competitive species system. AB - To address how species interactions, dispersal and environmental disturbances interplay to affect the spatial distribution and diversity of species, we present a compartment model in which multiple species undergo competitive interaction of Lotka-Volterra type in a patchy environment arranged in a square lattice. Dispersal of species occurs between adjacent patches. Disturbances are periodically imposed on a central part of the environment in a belt-like block or an island-like block of various sizes where each species is killed for a certain time interval and then allowed to recover for the rest of a disturbance cycle. We deal with a case in which the local population dynamics within each patch is analytically determinable and has multiple locally stable equilibrium states in the absence of environmental disturbance. We further assume a trade-off between the reproductive rate of species and its dispersal ability. With these settings, we numerically examine how the spatio-temporal distributions of species are affected by changes in the pattern, size and duration of disturbances. The results demonstrate that: (1) in the undisturbed area, environmental disturbances could generate spatially segregated distributions of species; (2) in the disturbed area, species with higher dispersal abilities quickly invade and preferentially recover their population during the post-disturbance period, being temporarily relieved of competition from other species. These mechanisms collectively lead to increased species diversity in the whole habitat, functioning best when both the size and duration of disturbances are intermediate. In particular, the belt-like disturbance is more effective than the island-like disturbance in sustaining spatial heterogeneity for a wider range of duration of disturbance. PMID- 12079364 TI - Mutations that reduce aggregation of the Alzheimer's Abeta42 peptide: an unbiased search for the sequence determinants of Abeta amyloidogenesis. AB - The primary component of amyloid plaque in the brains of Alzheimer's patients is the 42 residue amyloid-beta-peptide (Abeta42). Although the amino acid residue sequence of Abeta42 is known, the molecular determinants of Abeta amyloidogenesis have not been elucidated. To facilitate an unbiased search for the sequence determinants of Abeta aggregation, we developed a genetic screen that couples a readily observable phenotype in E. coli to the ability of a mutation in Abeta42 to reduce aggregation. The screen is based on our finding that fusions of the wild-type Abeta42 sequence to green fluorescent protein (GFP) form insoluble aggregates in which GFP is inactive. Cells expressing such fusions do not fluoresce. To isolate variants of Abeta42 with reduced tendencies to aggregate, we constructed and screened libraries of Abeta42-GFP fusions in which the sequence of Abeta42 was mutated randomly. Cells expressing GFP fusions to soluble (non-aggregating) variants of Abeta42 exhibit green fluorescence. Implementation of this screen enabled the isolation of 36 variants of Abeta42 with reduced tendencies to aggregate. The sequences of most of these variants are consistent with previous models implicating hydrophobic regions as determinants of Abeta42 aggregation. Some of the variants, however, contain amino acid substitutions not implicated in pre-existing models of Abeta amyloidogenesis. PMID- 12079367 TI - A two-component nucleation model of protein hydrophobicity. AB - A fundamental characteristic of soluble globular protein structure is a hydrophobic core and protein exterior comprised predominantly of hydrophilic residues. This distribution of amino acid residue hydrophobicity, from protein interior to exterior, has recently been profiled with the use of hydrophobic moments. The calculations enable comparison of the radial hydrophobicity distribution of different proteins and had revealed two features common to 30 proteins of diverse size and structure. One, a global feature, is the overall shape of the second-order ellipsoidal hydrophobic moment. The second, a specific feature, is a quasi-invariant hydrophobic-ratio of distances. Both features are dependent upon the rates of increase, from protein interior to exterior, of the accumulated numbers of hydrophobic and hydrophilic amino acid residues. These rates can be simulated simply with a two-component nucleation model of protein hydrophobicity. The model provides insight into the origin of the shape of the observed hydrophobic moment profiles and of the observed range of hydrophobic ratios. Consistent with observation, it is shown that a relatively wide range of hydrophobic and hydrophilic rates of increase yield a relatively narrow range of hydrophobic ratios. Furthermore, the model identifies one factor, the decrease in residue density with increasing distance from the protein interior, that is critical in providing the range of values that is comparable with the observed range. PMID- 12079369 TI - Molecular model for limb development. AB - The paper proposes a possible molecular mechanism of cell fate specification during limb development. The mechanism is based on a step-by-step activation of cell determinants, which occurs at the transcriptional level. Activation of the next determinant (or reactivation of the previous one) occurs only after the next cell division, only one determinant being activated (reactivated) in each cell cycle. This mechanism may be a basis for both proximodistal and anteroposterior specification of cell fates. As a result of this mechanism, the limb bud mesoderm proves to be divided into cell areas. Cells of the specific area would be condensed to form the specific cartilaginous element. PMID- 12079368 TI - A mathematical model of pattern formation in the vascular cambium of trees. AB - The beautiful patterns apparent in wood grain have their origin in the alignment of fusiform initial cells in the vascular cambium of trees. We develop a mathematical model to describe the orientation of fusiform initial cells, and their interaction with the plant hormone indole-3-acetic acid (auxin). The model incorporates the following four assumptions: (1) auxin is actively transported parallel to the long axis of the initials, (2) auxin diffuses perpendicular to the long axis of the initials, (3) the initials tend to orient parallel to the flux of auxin through the cambium, and (4) adjacent initials tend to orient parallel to one another. Each assumption is justified on the basis of available evidence and cast in mathematical form. Our main result is a pair of nonlinear differential equations that describe the coupling between the distribution of auxin in the cambium and the orientation of fusiform initials. Numerical solutions to the equations show qualitative resemblance to the wood grain patterns observed at branch junctions, wounds and knots, and topological defects. PMID- 12079370 TI - The role of stoichiometric analysis in studies of metabolism: an example. AB - Stoichiometric analysis uses matrix algebra to deduce the constraints implicit in metabolic networks. When applied to simple networks, it can often give the impression of being an unnecessarily complicated way of arriving at information that is obvious from inspection, for example, that the sum of the concentrations of the adenine nucleotides is constant. Applied to a more complicated example, that of glycolysis in Trypanosoma brucei, it yields information that is far from obvious and may have importance for developing therapeutic ways of eliminating this parasite. Even in simplified form, the network contains nine reactions or transport steps involving 11 metabolites. This immediately shows that there must be at least two stoichiometric constraints, and indeed two can be recognized by inspection: conservation of adenine nucleotides and conservation of the two forms of NAD. There is, however, a third, which involves eight different phosphorylated intermediates in non-obvious combinations and is very difficult to recognize by inspection. It is also difficult to recognize by inspection that no fourth stoichiometric constraint exists. Gaussian elimination provides a systematic way of analysing a network in such a way that all the stoichiometric relationships that it contains emerge automatically. PMID- 12079371 TI - Comparative study of circadian clock models, in search of processes promoting oscillation. AB - We study simple models for circadian rhythm, and examine the condition in which the equilibrium is unstable, generating a sustained oscillation. In the models, a clock gene(s) is transcribed to produce mRNAs, which are translated to produce proteins that suppress the transcription of the clock gene(s). First, using a Lyapunov function, we prove under very general conditions that two-variable models cannot generate a stable oscillation, implying that additional structures are needed for the model to generate a sustainable rhythm. By comparing several models of different complexities using the Routh-Hurwitz criteria of stability, we show that a sustained oscillation is more likely to occur if the cell is compartmentalized and the proteins need to be transported from the cytosol to the nucleus, if the proteins have to be modified before entering the nucleus, if the kinetics of transcription inhibition or the transport to the nucleus have cooperativity with a nonlinear dependence on the substrate concentration, or if the products of two clock genes form a heterodimer that suppresses both of their own genes. We discuss the implications of these results. PMID- 12079372 TI - The regulatory anatomy of honeybee lifespan. AB - Honeybee workers (Apis mellifera) may be classified as either short-lived summer bees or long-lived winter bees in temperate zones. The protein status appears to be a major determinant of honeybee lifespan, and the lipoprotein vitellogenin seems to play a crucial role. Here, we give a review of the role of the vitellogenin in honeybee workers, and present a data-driven mathematical model describing the dynamics of this representative protein in the individual bee as a function of its task profile under various regimes. The results support the hypothesis that vitellogenin is a true storage protein that is utilized for various metabolic purposes including the synthesis of brood food. Except for workers having been foragers for many days, they also suggest that the previous life histories of workers do not constrain them from becoming winter bees as long as they get ample food and time to build up their protein reserves before wintering. The results also indicate that it may not be necessary to introduce the ovary as a storage organ for vitellogenin in order to generate normal winter bees. The insights gained from these results are then discussed in a broader gerontological and life history context. Remarkably similar features concerning regulation of ageing in Caenorhabditis elegans, Drosophila melanogaster and honeybees are pointed out and discussed. Furthermore, we show that in contrast to the "mutation accumulation" and the "antagonistic pleiotropy" evolutionary theories of ageing, the "disposable soma" theory is capable of explaining the bimodal longevity distribution of honeybees when interpreted in a group selection context. Finally, by showing that depletion of nutrient stores can be actively controlled by pathways connected to regulation of ageing, we strengthen the claim that age-based division of labour, with performance of risky tasks delayed until late in life by workers with depleted nutrient stores, may have evolved as an energy-saving mechanism in insect colonies. PMID- 12079373 TI - Positive and negative feedback: striking a balance between necessary antagonists. AB - Most biological regulation systems comprise feedback circuits as crucial components. Negative feedback circuits have been well understood for a very long time; indeed, their understanding has been the basis for the engineering of cybernetic machines exhibiting stable behaviour. The importance of positive feedback circuits, considered as "vicious circles", has however been underestimated. In this article, we give a demonstration based on degree theory for vector fields of the conjecture, made by Rene Thomas, that the presence of positive feedback circuits is a necessary condition for autonomous differential systems, covering a wide class of biologically relevant systems, to possess multiple steady states. We also show ways to derive constraints on the weights of positive and negative feedback circuits. These qualitative and quantitative results provide, respectively, structural constraints (i.e. related to the interaction graph) and numerical constraints (i.e. related to the magnitudes of the interactions) on systems exhibiting complex behaviours, and should make it easier to reverse-engineer the interaction networks animating those systems on the basis of partial, sometimes unreliable, experimental data. We illustrate these concepts on a model multistable switch, in the context of cellular differentiation, showing a requirement for sufficient cooperativity. Further developments are expected in the discovery and modelling of regulatory networks in general, and in the interpretation of bio-array hybridization and proteomics experiments in particular. PMID- 12079374 TI - Mechanical implications of the domain structure of fiber-forming collagens: comparison of the molecular and fibrillar flexibilities of the alpha1-chains found in types I-III collagen. AB - Fibrillar collagens store, transmit and dissipate elastic energy during tensile deformation. Results of previous studies suggest that the collagen molecule is made up of alternating rigid and flexible domains, and extension of the flexible domains is associated with elastic energy storage. In this study, we model the flexibility of the alpha1-chains found in types I-III collagen molecules and microfibrils in order to understand the molecular basis of elastic energy storage in collagen fibers by analysing the areas under conformational plots for dipeptide sequences. Results of stereochemical modeling suggest that the collagen triple helix is made up of rigid and flexible domains that alternate with periods that are multiples of three amino acid residues. The relative flexibility of dipeptide sequences found in the flexible regions is about a factor of five higher than that found for the flexibility of the rigid regions, and the flexibility of types II and III collagen molecules appears to be higher than that found for the type I collagen molecule. The different collagen alpha1-chains were compared by correlating the flexibilities. The results suggest that the flexibilities of the alpha1-chains of types I and III collagen are more closely related than the flexibilities of the alpha1-chains in types I and II and II and III collagen. The flexible domains found in the alpha1-chains of types I-III collagen were found to be conserved in the microfibril and had periods of about 15 amino acid residues and multiples thereof. The flexibility profiles of types I and II collagen microfibrils were found to be more highly correlated than those for types I and III and II and III. These results suggest that the domain structure of the alpha1-chains found in types I-III collagen is an efficient means for storage of elastic energy during stretching while preserving the triple helical structure of the overall molecule. It is proposed that all collagens that form fibers are designed to act as storage elements for elastic energy. The function of fibers rich in type I collagen is to store and then transmit this energy while fibers rich in types II and III collagen may store and then reflect elastic energy for dissipation through viscous fibrillar slippage. Impaired elastic energy storage by extracellular matrices may lead to cellular damage and changes in signaling by mechanochemical transduction at the extracellular matrix cell interface. PMID- 12079375 TI - The cytochrome c folding landscape revealed by electron-transfer kinetics. AB - We have investigated the folding energy landscape of cytochrome c by exploiting the widely different electron-transfer (ET) reactivities of buried and exposed Zn(II)-substituted hemes. An electronically excited Zn-porphyrin in guanidine hydrochloride denatured Zn-substituted cytochrome c (Zn-cyt c) reduces ruthenium(III) hexaammine about ten times faster than when embedded in the fully folded protein. Measurements of ET kinetics during Zn-cyt c folding reveal a burst intermediate in which one-third of the ensemble has a protected Zn porphyrin and slow ET kinetics; the remaining fraction exhibits fast ET characteristic of a solvent-exposed redox cofactor. The ET data show that, under solvent conditions favoring the folded protein, collapsed non-native structures are not substantially more stable than extended conformations, and that the two populations interchange rapidly. Most of the folding free energy, then, is released when compact structures evolve into the native fold. PMID- 12079376 TI - Equilibrium folding and stability of myotrophin: a model ankyrin repeat protein. AB - Proteins containing stretches of repeating amino acid sequences are prevalent throughout nature, yet little is known about the general folding and assembly mechanisms of these systems. Here we propose myotrophin as a model system to study the folding of ankyrin repeat proteins. Myotrophin is folded over a large pH range and is soluble at high concentrations. Thermal and urea denaturation studies show that the protein displays cooperative two-state folding properties despite its modular nature. Taken together with previous studies on other ankyrin repeat proteins, our data suggest that the two-state folding pathway may be characteristic of ankyrin repeat proteins and other integrated alpha-helical repeat proteins in general. PMID- 12079378 TI - Crystal structure of the potassium form of an Oxytricha nova G-quadruplex. AB - The crystal structures of the potassium-containing quadruplex formed from the Oxytricha nova sequence d(GGGGTTTTGGGG) are reported, in two space groups, the orthorhombic P2(1)2(1)2(1) and the trigonal P3(2)21, which diffract to 2.0 A and 1.49 A, respectively. The orthorhombic form contains two independent quadruplexes in the asymmetric unit, and the trigonal form contains one. All three of these quadruplexes adopt an identical fold, with two strands forming an antiparallel diagonal arrangement. This is identical with that observed previously in NMR studies of the native sodium and potassium forms, and a crystallographic analysis of it complexed with an O. nova protein. The present analysis demonstrates that the native structure is the same in solution and in the crystalline state and, moreover, that the nature of the counter-ion does not affect the overall fold of this quadruplex. The analysis corrects an earlier crystallographic study of this quadruplex. The conformation of the tetra-thymine loop is described in detail, which involves the third thymine base folding back to interact with the first thymine base. The water networks in the grooves and loops are described and, in particular, the ability of water molecules to form a continuous spine of hydration in the narrow groove is detailed. Each quadruplex has five potassium ions organised in a linear channel, with square antiprismatic coordination to each ion from oxygen atoms. PMID- 12079377 TI - The ATP-operated clamp of human DNA topoisomerase IIalpha: hyperstimulation of ATPase by "piggy-back" binding. AB - We have constructed a series of clones encoding N-terminal fragments of human DNA topoisomerase IIalpha. All fragments exhibit DNA-dependent ATPase activity. Fragment 1-420 shows hyperbolic dependence of ATPase on DNA concentration, whereas fragment 1-453 shows hyperstimulation at low ratios of DNA to enzyme, a phenomenon found previously with the full-length enzyme. The minimum length of DNA found to stimulate the ATPase activity was approximately 10 bp; fragments >or=32 bp manifest the hyperstimulation phenomenon. Molecular mass studies show that fragment 1-453 is a monomer in the absence of nucleotides and a dimer in the presence of nucleotide triphosphate. The results are consistent with the role of the N-terminal domain of topoisomerase II as an ATP-operated clamp that dimerises in the presence of ATP. The hyperstimulation effect can be interpreted in terms of a "piggy-back binding" model for protein-DNA interaction. PMID- 12079379 TI - Expression, purification and characterisation of full-length histidine protein kinase RegB from Rhodobacter sphaeroides. AB - The global redox switch between aerobic and anaerobic growth in Rhodobacter sphaeroides is controlled by the RegA/RegB two-component system, in which RegB is the integral membrane histidine protein kinase, and RegA is the cytosolic response regulator. Despite the global regulatory importance of this system and its many homologues, there have been no reported examples to date of heterologous expression of full-length RegB or any histidine protein kinases. Here, we report the amplified expression of full-length functional His-tagged RegB in Escherichia coli, its purification, and characterisation of its properties. Both the membrane bound and purified solubilised RegB protein demonstrate autophosphorylation activity, and the purified protein autophosphorylates at the same rate under both aerobic and anaerobic conditions confirming that an additional regulator is required to control/inhibit autophosphorylation. The intact protein has similar activity to previously characterised soluble forms, but is dephosphorylated more rapidly than the soluble form (half-life ca 30 minutes) demonstrating that the transmembrane segment present in the full-length RegB may be an important regulator of RegB activity. Phosphotransfer from RegB to RegA (overexpressed and purified from E. coli) by RegB is very rapid, as has been reported for the soluble domain. Dephosphorylation of active RegA by full-length RegB has a rate similar to that observed previously for soluble RegB. PMID- 12079380 TI - First structural evidence of a specific inhibition of phospholipase A2 by alpha tocopherol (vitamin E) and its implications in inflammation: crystal structure of the complex formed between phospholipase A2 and alpha-tocopherol at 1.8 A resolution. AB - This is the first structural evidence of alpha-tocopherol (alpha-TP) as a possible candidate against inflammation, as it inhibits phospholipase A2 specifically and effectively. The crystal structure of the complex formed between Vipera russelli phospholipase A2 and alpha-tocopherol has been determined and refined to a resolution of 1.8 A. The structure contains two molecules, A and B, of phospholipase A2 in the asymmetric unit, together with one alpha-tocopherol molecule, which is bound specifically to one of them. The phospholipase A2 molecules interact extensively with each other in the crystalline state. The two molecules were found in a stable association in the solution state as well, thus indicating their inherent tendency to remain together as a structural unit, leading to significant functional implications. In the crystal structure, the most important difference between the conformations of two molecules as a result of their association pertains to the orientation of Trp31. It may be noted that Trp31 is located at the mouth of the hydrophobic channel that forms the binding domain of the enzyme. The values of torsion angles (phi, psi, chi(1) and chi(2)) for both the backbone as well as for the side-chain of Trp31 in molecules A and B are -94 degrees, -30 degrees, -66 degrees, 116 degrees and -128 degrees, 170 degrees, -63 degrees, -81 degrees, respectively. The conformation of Trp31 in molecule A is suitable for binding, while that in B hinders the passage of the ligand to the binding site. Consequently, alpha-tocopherol is able to bind to molecule A only, while the binding site of molecule B contains three water molecules. In the complex, the aromatic moiety of alpha-tocopherol is placed in the large space at the active site of the enzyme, while the long hydrophobic channel in the enzyme is filled by hydrocarbon chain of alpha-tocopherol. The critical interactions between the enzyme and alpha-tocopherol are generated between the hydroxyl group of the six-membered ring of alpha-tocopherol and His48 N(delta1) and Asp49 O(delta1) as characteristic hydrogen bonds. The remaining part of alpha-tocopherol interacts extensively with the residues of the hydrophobic channel of the enzyme, giving rise to a number of hydrophobic interactions, resulting in the formation of a stable complex. PMID- 12079381 TI - Crystal structure of guanidinoacetate methyltransferase from rat liver: a model structure of protein arginine methyltransferase. AB - Guanidinoacetate methyltransferase (GAMT) is the enzyme that catalyzes the last step of creatine biosynthesis. The enzyme is found in abundance in the livers of all vertebrates. Recombinant rat liver GAMT has been crystallized with S adenosylhomocysteine (SAH), and the crystal structure has been determined at 2.5 A resolution. The 36 amino acid residues at the N terminus were cleaved during the purification and the truncated enzyme was crystallized. The truncated enzyme forms a dimer, and each subunit contains one SAH molecule in the active site. Arg220 of the partner subunit forms a pair of hydrogen bonds with Asp134 at the guanidinoacetate-binding site. On the basis of the crystal structure, site directed mutagenesis on Asp134, and chemical modification and limited proteolysis studies, we propose a catalytic mechanism of this enzyme. The truncated GAMT dimer structure can be seen as a ternary complex of protein arginine methyltransferase (one subunit) complexed with a protein substrate (the partner subunit) and the product SAH. Therefore, this structure provides insight into the structure and catalysis of protein arginine methyltransferases. PMID- 12079382 TI - Structure of porphobilinogen synthase from Pseudomonas aeruginosa in complex with 5-fluorolevulinic acid suggests a double Schiff base mechanism. AB - All natural tetrapyrroles, including hemes, chlorophylls and vitamin B12, share porphobilinogen (PBG) as a common precursor. Porphobilinogen synthase (PBGS) synthesizes PBG through the asymmetric condensation of two molecules of aminolevulinic acid (ALA). Crystal structures of PBGS from various sources confirm the presence of two distinct binding sites for each ALA molecule, termed A and P. We have solved the structure of the active-site variant D139N of the Mg2+-dependent PBGS from Pseudomonas aeruginosa in complex with the inhibitor 5 fluorolevulinic acid at high resolution. Uniquely, full occupancy of both substrate binding sites each by a single substrate-like molecule was observed. Both inhibitor molecules are covalently bound to two conserved, active-site lysine residues, Lys205 and Lys260, through Schiff bases. The active site now also contains a monovalent cation that may critically enhance enzymatic activity. Based on these structural data, we postulate a catalytic mechanism for P. aeruginosa PBGS initiated by a C-C bond formation between A and P-side ALA, followed by the formation of the intersubstrate Schiff base yielding the product PBG. PMID- 12079383 TI - Crystal structure of MabA from Mycobacterium tuberculosis, a reductase involved in long-chain fatty acid biosynthesis. AB - The fatty acid elongation system FAS-II is involved in the biosynthesis of mycolic acids, which are major and specific long-chain fatty acids of the cell envelope of Mycobacterium tuberculosis and other mycobacteria, including Mycobacterium smegmatis. The protein MabA, also named FabG1, has been shown recently to be part of FAS-II and to catalyse the NADPH-specific reduction of long chain beta-ketoacyl derivatives. This activity corresponds to the second step of an FAS-II elongation round. FAS-II is inhibited by the antituberculous drug isoniazid through the inhibition of the 2-trans-enoyl-acyl carrier protein reductase InhA. Thus, the other enzymes making up this enzymatic complex represent potential targets for designing new antituberculous drugs. The crystal structure of the apo-form MabA was solved to 2.03 A resolution by molecular replacement. MabA is tetrameric and shares the conserved fold of the short-chain dehydrogenases/reductases (SDRs). However, it exhibits some significant local rearrangements of the active-site loops in the absence of a cofactor, particularly the beta5-alpha5 region carrying the unique tryptophan residue, in agreement with previous fluorescence spectroscopy data. A similar conformation has been observed in the beta-ketoacyl reductase from Escherichia coli and the distantly related dehydratase. The distinctive enzymatic and structural properties of MabA are discussed in view of its crystal structure and that of related enzymes. PMID- 12079384 TI - Crystal structures of the Met148Leu and Ser86Asp mutants of rusticyanin from Thiobacillus ferrooxidans: insights into the structural relationship with the cupredoxins and the multi copper proteins. AB - The crystal structures of the Met148Leu and Ser86Asp mutants of rusticyanin are presented at 1.82 and 1.65 A resolution, respectively. Both of these structures have two molecules in the asymmetric unit compared to the one present in the crystal form of the native protein. This provides an opportunity to investigate intramolecular electron transfer pathways in rusticyanin. The redox potential of the Met148Leu mutant ( approximately 800 mV) is elevated compared to that of the native protein ( approximately 670 mV at pH 3.2) while that of the Ser86Asp mutant ( approximately 623 mV at pH 3.2) is decreased. The effect of the Ser86Asp mutation on the hydrogen bonding near the type 1 Cu site is discussed and hence its role in determining acid stability is examined. The type 1 Cu site of Met148Leu mimics the structural and biochemical characteristics of those found in domain II of ceruloplasmin and fungal laccase. Moreover, the native rusticyanin's cupredoxin core and the type 1 Cu site closely resemble those found in ascorbate oxidase and nitrite reductase. Structure based phylogenetic trees have been re examined in view of the additional structural data on rusticyanin and fungal laccase. We confirm that rusticyanin is in the same class as nitrite reductase domain 2, laccase domain 3 and ceruloplasmin domains 2, 4 and 6. PMID- 12079385 TI - Additional binding sites for anionic phospholipids and calcium ions in the crystal structures of complexes of the C2 domain of protein kinase calpha. AB - The C2 domain of protein kinase Calpha (PKCalpha) corresponds to the regulatory sequence motif, found in a large variety of membrane trafficking and signal transduction proteins, that mediates the recruitment of proteins by phospholipid membranes. In the PKCalpha isoenzyme, the Ca2+-dependent binding to membranes is highly specific to 1,2-sn-phosphatidyl-l-serine. Intrinsic Ca2+ binding tends to be of low affinity and non-cooperative, while phospholipid membranes enhance the overall affinity of Ca2+ and convert it into cooperative binding. The crystal structure of a ternary complex of the PKCalpha-C2 domain showed the binding of two calcium ions and of one 1,2-dicaproyl-sn-phosphatidyl-l-serine (DCPS) molecule that was coordinated directly to one of the calcium ions. The structures of the C2 domain of PKCalpha crystallised in the presence of Ca2+ with either 1,2 diacetyl-sn-phosphatidyl-l-serine (DAPS) or 1,2-dicaproyl-sn-phosphatidic acid (DCPA) have now been determined and refined at 1.9 A and at 2.0 A, respectively. DAPS, a phospholipid with short hydrocarbon chains, was expected to facilitate the accommodation of the phospholipid ligand inside the Ca2+-binding pocket. DCPA, with a phosphatidic acid (PA) head group, was used to investigate the preference for phospholipids with phosphatidyl-l-serine (PS) head groups. The two structures determined show the presence of an additional binding site for anionic phospholipids in the vicinity of the conserved lysine-rich cluster. Site-directed mutagenesis, on the lysine residues from this cluster that interact directly with the phospholipid, revealed a substantial decrease in C2 domain binding to vesicles when concentrations of either PS or PA were increased in the absence of Ca2+. In the complex of the C2 domain with DAPS a third Ca2+, which binds an extra phosphate group, was identified in the calcium-binding regions (CBRs). The interplay between calcium ions and phosphate groups or phospholipid molecules in the C2 domain of PKCalpha is supported by the specificity and spatial organisation of the binding sites in the domain and by the variable occupancies of ligands found in the different crystal structures. Implications for PKCalpha activity of these structural results, in particular at the level of the binding affinity of the C2 domain to membranes, are discussed. PMID- 12079386 TI - The structure of an allosamidin complex with the Coccidioides immitis chitinase defines a role for a second acid residue in substrate-assisted mechanism. AB - Allosamidin is a known inhibitor of class 18 chitinases. We show that allosamidin is a competitive inhibitor of the fungal chitinase CiX1 from Coccidioides immitis, with a K(i) of 60 nM. We report the X-ray structure of the complex and show that upon inhibitor binding the side-chain of Asp169 rotates to form an ion pair with the oxazolinium cation. The mechanism of action is thought to involve protonation of the leaving group by Glu171 and substrate assistance by the sugar acetamido moiety to form an oxazoline-like intermediate. We converted both amino acid residues to the corresponding amide and found that each mutation effectively abolishes enzyme activity. X-ray structures show the mutant enzymes retain the basic wild-type structure and that the loss of mutant activity is due to their altered chemical properties. The high affinity of allosamidin, and its similarity to the putative reaction intermediate, suggests it is a transition state analog. This helps validate our contention that the role of Asp169 is to electrostatically stabilize the reaction transition state. PMID- 12079387 TI - A novel combination of two classic catalytic schemes. AB - The crystal structure of an alkaline Bacillus cellulase catalytic core, from glucoside hydrolase family 5, reveals a novel combination of the catalytic machinery of two classic textbook enzymes. The enzyme has the expected two glutamate residues in close proximity to one another in the active-site that are typical of retaining cellulases. However, the proton donor, glutamate 139 is also unexpectedly a member of a serine-histidine-glutamate catalytic triad, forming a novel combination of catalytic machineries. Structure and sequence analysis of glucoside hydrolase family 5 reveal that the triad is highly conserved, but with variations at the equivalent of the serine position. We speculate that the purpose of this novel catalytic triad is to control the protonation of the acid/base glutamate, facilitating the first step of the catalytic reaction, protonation of the substrate, by the proton donor glutamate. If correct, this will be a novel use for a catalytic triad. PMID- 12079388 TI - High pressure NMR reveals that apomyoglobin is an equilibrium mixture from the native to the unfolded. AB - Pressure-induced reversible conformational changes of sperm whale apomyoglobin have been studied between 30 bar and 3000 bar on individual residue basis by utilizing 1H/15N hetero nuclear single-quantum coherence two-dimensional NMR spectroscopy at pH 6.0 and 35 degrees C. Apomyoglobin showed a series of pressure dependent NMR spectra as a function of pressure, assignable to the native (N), intermediates (I), molten globule (MG) and unfolded (U) conformers. At 30 bar, the native fold (N) shows disorder only in the F helix. Between 500 bar and 1200 bar, a series of locally disordered conformers I are produced, in which local disorder occurs in the C helix, the CD loop, the G helix and part of the H helix. At 2000 bar, most cross-peaks exhibit severe line-broadening, suggesting the formation of a molten globule, but at 3000 bar all the cross-peaks reappear, showing that the molten globule turns into a well-hydrated, mobile unfolded conformation U. Since all the spectral changes were reversible with pressure, apomyoglobin is considered to exist as an equilibrium mixture of the N, I, MG and U conformers at all pressures. MG is situated at 2.4+/-(0.1) kcal/mol above N at 1 bar and the unfolding transition from the combined N-I state to MG is accompanied by a loss of partial molar volume by 75+/-(3) ml/mol. On the basis of these observations, we postulate a theorem that the partial molar volume of a protein decreases in parallel with the loss of its conformational order. PMID- 12079389 TI - Solution structure of the single-domain prolyl cis/trans isomerase PIN1At from Arabidopsis thaliana. AB - The 119-amino acid residue prolyl cis/trans isomerase from Arabidopsis thaliana (PIN1At) is similar to the catalytic domain of the human hPIN1. However, PIN1At lacks the N-terminal WW domain that appears to be essential for the hPIN1 function. Here, the solution structure of PIN1At was determined by three dimensional nuclear magnetic resonance spectroscopy. The PIN1At fold could be superimposed on that of the catalytic domain of hPIN1 and had a 19 residue flexible loop located between strand beta1 and helix alpha1. The dynamical features of this beta1/alpha1-loop, which are characteristic for a region involved in protein-protein interactions, led to exchange broadening in the NMR spectra. When sodium sulfate salt was added to the protein sample, the beta1/alpha1 loop was stabilized and, hence, a complete backbone resonance assignment was obtained. Previously, with a phospho-Cdc25 peptide as substrate, PIN1At had been shown to catalyze the phosphoserine/phosphothreonine prolyl cis/trans isomerization specifically. To map the catalytic site of PIN1At, the phospho-Cdc25 peptide or sodium sulfate salt was added in excess to the protein and chemical shift changes in the backbone amide protons were monitored in the (1)H(N)-(15)N heteronuclear single quantum coherence spectrum. The peptide caused perturbations in the loops between helix alpha4 and strand beta3, between strands beta3 and beta4, in the alpha3 helix, and in the beta1/alpha1 loop. The amide groups of the residues Arg21 and Arg22 showed large chemical shift perturbations upon phospho-Cdc25 peptide or sulfate addition. We conclude that this basic cluster formed by Arg21 and Arg22, both located in the beta1/alpha1 loop, is homologous to that found in the hPIN1 crystal structure (Arg68 and Arg69), which also is involved in sulfate ion binding. We showed that the sulfate group competed for the interaction between PIN1At and the phospho-Cdc25 peptide. In the absence of the WW domain, three hydrophobic residues (Ile33, Ile34, and Leu35) located in the long flexible loop and specific for the plant PIN-type peptidyl prolyl cis/trans isomerases (PPIases) could be an additional interaction site in PIN1At. However, phospho-peptide addition did not affect the resonances of these residues significantly. Electrostatic potential calculations revealed a negatively charged area not found in hPIN1 on the PIN1At molecular surface, which corresponds to the surface shielded by the WW domain in hPIN1. Based on our experimental results and the molecular specificities of the PIN1At enzyme, functional implications of the lack of WW domains in this plant PIN-type PPIase will be discussed. PMID- 12079390 TI - Decreased thermodynamic stability as a crucial factor for familial amyloidotic polyneuropathy. AB - A single mutation in the wild-type transthyretin (WT TTR) such as V30M causes a familial amyloidotic polyneuropathy disease. Comparison of the three-dimensional crystal structures of WT and V30M does not tell much about the reason. High pressure NMR revealed that at neutral pH both WT and V30M exist as equilibrium between the native tetramer and the dissociated/unfolded monomer. The native tetramer is highly stable in WT (deltaG(0)=104 kJ/mol at 37 degrees C, pH 7.1), but the stability is significantly reduced in V30M (deltadeltaG(0)=-18 kJ/mol), increasing the fraction of the unfolded monomer by a 1000-fold. Significant reduction of thermodynamic stability of WT TTR by mutation could be a crucial factor for familial amyloidotic polyneuropathy. PMID- 12079391 TI - Thermodynamic consequences of burial of polar and non-polar amino acid residues in the protein interior. AB - Effects of amino acid substitutions at four fully buried sites of the ubiquitin molecule on the thermodynamic parameters (enthalpy, Gibbs energy) of unfolding were evaluated experimentally using differential scanning calorimetry. The same set of substitutions has been incorporated at each of four sites. These substitutions have been designed to perturb packing (van der Waals) interactions, hydration, and/or hydrogen bonding. From the analysis of the thermodynamic parameters for these ubiquitin variants we conclude that: (i) packing of non polar groups in the protein interior is favorable and is largely defined by a favorable enthalpy of van der Waals interactions. The removal of one methylene group from the protein interior will destabilize a protein by approximately 5 kJ/mol, and will decrease the enthalpy of a protein by 12 kJ/mol. (ii) Burial of polar groups in the non-polar interior of a protein is highly destabilizing, and the degree of destabilization depends on the relative polarity of this group. For example, burial of Thr side-chain in the non-polar interior will be less destabilizing than burial of Asn side-chain. This decrease in stability is defined by a large enthalpy of dehydration of polar groups upon burial. (iii) The destabilizing effect of dehydration of polar groups upon burial can be compensated if these buried polar groups form hydrogen bonding. The enthalpy of this hydrogen bonding will compensate for the unfavorable dehydration energy and as a result the effect will be energetically neutral or even slightly stabilizing. PMID- 12079392 TI - Computational analysis of the chiral action of type II DNA topoisomerases. AB - It was found recently that bacterial type II DNA topoisomerase, topo IV, is much more efficient in relaxing (+) DNA supercoiling than (-) supercoiling. This means that the DNA-enzyme complex is chiral. This chirality can appear upon binding the first segment that participates in the strand passing reaction (G segment) or only after the second segment (T segment) joins the complex. The former possibility is analyzed here. We assume that upon binding the enzyme, the G segment forms a part of left-handed helical turn. This model is an extension of the hairpin model introduced earlier to explain simplification of DNA topology by these enzymes. Using statistical-mechanical simulation of DNA properties, we estimated different consequences of the model: (1) relative rates of relaxation of (+) and (-) supercoiling by the enzyme; (2) the distribution of positions of the G segment in supercoiled molecules; (3) steady-state distribution of knots in circular molecules created by the topoisomerase; (4) the variance of topoisomer distribution created by the enzyme; (5) the effect of (+) and (-) supercoiling on the binding topo II with G segment. The simulation results are capable of explaining nearly all available experimental data, at least semiquantitatively. A few predictions obtained in the model analysis can be tested experimentally. PMID- 12079393 TI - Predicting changes in the stability of proteins and protein complexes: a study of more than 1000 mutations. AB - We have developed a computer algorithm, FOLDEF (for FOLD-X energy function), to provide a fast and quantitative estimation of the importance of the interactions contributing to the stability of proteins and protein complexes. The predictive power of FOLDEF was tested on a very large set of point mutants (1088 mutants) spanning most of the structural environments found in proteins. FOLDEF uses a full atomic description of the structure of the proteins. The different energy terms taken into account in FOLDEF have been weighted using empirical data obtained from protein engineering experiments. First, we considered a training database of 339 mutants in nine different proteins and optimised the set of parameters and weighting factors that best accounted for the changes in stability of the mutants. The predictive power of the method was then tested using a blind test mutant database of 667 mutants, as well as a database of 82 protein-protein complex mutants. The global correlation obtained for 95 % of the entire mutant database (1030 mutants) is 0.83 with a standard deviation of 0.81 kcal mol(-1) and a slope of 0.76. The present energy function uses a minimum of computational resources and can therefore easily be used in protein design algorithms, and in the field of protein structure and folding pathways prediction where one requires a fast and accurate energy function. FOLDEF is available via a web-interface at http://fold-x.embl-heidelberg.de PMID- 12079394 TI - Protein folding kinetics beyond the phi value: using multiple amino acid substitutions to investigate the structure of the SH3 domain folding transition state. AB - The SH3 domain folding transition state structure contains two well-ordered turn regions, known as the diverging turn and the distal loop. In the Src SH3 domain transition state, these regions are stabilized by a hydrogen bond between Glu30 in the diverging turn and Ser47 in the distal loop. We have examined the effects on folding kinetics of amino acid substitutions at the homologous positions (Glu24 and Ser41) in the Fyn SH3 domain. In contrast to most other folding kinetics studies which have focused primarily on non-disruptive substitutions with Ala or Gly, here we have examined the effects of substitutions with diverse amino acid residues. Using this approach, we demonstrate that the transition state structure is generally tolerant to amino acid substitutions. We also uncover a unique role for Ser at position 41 in facilitating folding of the distal loop, which can only be replicated by Asp at the same position. Both these residues appear to accelerate folding through the formation of short-range side chain to backbone hydrogen bonds. The folding of the diverging turn region is shown to be driven primarily by local interactions. The diverging turn and distal loop regions are found to interact in the transition state structure, but only in the context of particular mutant backgrounds. This work demonstrates that studying the effects of a variety of amino acid substitutions on protein folding kinetics can provide unique insights into folding mechanisms which cannot be obtained by standard Phi value analysis. PMID- 12079395 TI - The systematic substitutions around the conserved charged residues of the cytoplasmic loop of Na+-driven flagellar motor component PomA. AB - PomA, a homolog of MotA in the H+-driven flagellar motor, is an essential component for torque generation in the Na+-driven flagellar motor. Previous studies suggested that two charged residues, R90 and E98, which are in the single cytoplasmic loop of MotA, are directly involved in this process. These residues are conserved in PomA of Vibrio alginolyticus as R88 and E96, respectively. To explore the role of these charged residues in the Na+-driven motor, we replaced them with other amino acids. However, unlike in the H+-driven motor, both of the single and the double PomA mutants were functional. Several other positively and negatively charged residues near R88 and E96, namely K89, E97 and E99, were neutralized. Motility was retained in a strain producing the R88A/K89A/E96Q/E97Q/E99Q (AAQQQ) PomA protein. The swimming speed of the AAQQQ strain was as fast as that of the wild-type PomA strain, but the direction of motor rotation was abnormally counterclockwise-biased. We could, however, isolate non-motile or poorly motile mutants when certain charged residues in PomA were reversed or neutralized. The charged residues at positions 88-99 of PomA may not be essential for torque generation in the Na+-driven motor and might play a role in motor function different from that of the equivalent residues of the H+-driven motor. PMID- 12079396 TI - Comprehensive functional maps of the antigen-binding site of an anti-ErbB2 antibody obtained with shotgun scanning mutagenesis. AB - Shotgun scanning combinatorial mutagenesis was used to study the antigen-binding site of Fab2C4, a humanized monoclonal antibody fragment that binds to the extracellular domain of the human oncogene product ErbB2. Essentially all the residues in the Fab2C4 complementarity determining regions (CDRs) were alanine scanned using phage-displayed libraries that preferentially allowed side-chains to vary as the wild-type or alanine. A separate homolog-scan was performed using libraries that allowed side-chains to vary only as the wild-type or a similar amino acid residue. Following binding selections to isolate functional clones, DNA sequencing was used to determine the wild-type/mutant ratios at each varied position, and these ratios were used to assess the contributions of each side chain to antigen binding. The alanine-scan revealed that most of the side-chains that contribute to antigen binding are located in the heavy chain, and the Fab2C4 three-dimensional structure revealed that these residues fall into two groups. The first group consists of solvent-exposed residues which likely make energetically favorable contacts with the antigen and thus comprise the functional-binding epitope. The second group consists of buried residues with side-chains that pack against other CDR residues and apparently act as scaffolding to maintain the functional epitope in a binding-competent conformation. The homolog-scan involved subtle mutations, and as a result, only a subset of the side-chains that were intolerant to alanine substitutions were also intolerant to homologous substitutions. In particular, the 610 A2 functional epitope surface revealed by alanine-scanning shrunk to only 369 A2 when mapped with homologous substitutions, suggesting that this smaller subset of side-chains may be involved in more precise contacts with the antigen. The results validate shotgun scanning as a rapid and accurate method for determining the functional contributions of individual side-chains involved in protein-protein interactions. PMID- 12079398 TI - Expression pattern of a mini human PrP gene promoter in transgenic mice. AB - The prion protein is central to the pathogenesis of prion diseases, although its exact function remains unclear. Although transgenic mice have been widely utilised in prion research, their PrP expression patterns have not been characterised in detail. We have studied the developmental temporal and spatial expression of a 214-bp mini human PrP promoter in transgenic mice. Transgene expression is first detected at embryonic day 12.5, a day earlier than previously reported for endogenous mouse gene by in situ hybridization. The general expression pattern closely mirrors that of the endogenous mouse PrP gene, such that this small and clearly defined transgene cassette can replace the need to use large cosmid based vectors for transgenetic modeling of human and animal prion disease. PMID- 12079399 TI - Overexpression of wild type but not an FAD mutant presenilin-1 promotes neurogenesis in the hippocampus of adult mice. AB - Mutations in the presenilin-1 (PS-1) gene are one cause of familial Alzheimer's disease (FAD). However, the functions of the PS-1 protein as well as how PS-1 mutations cause FAD are incompletely understood. Here we investigated if neuronal overexpression of wild-type or FAD mutant PS-1 in transgenic mice affects neurogenesis in the hippocampus of adult animals. We show that either a wild-type or an FAD mutant PS-1 transgene reduces the number of neural progenitors in the dentate gyrus. However, the wild-type, but not the FAD mutant PS-1 promoted the survival and differentiation of progenitors leading to more immature granule cell neurons being generated in PS-1 wild type expressing animals. These studies suggest that PS-1 plays a role in regulating neurogenesis in adult hippocampus and that FAD mutants may have deleterious properties independent of their effects on amyloid deposition. PMID- 12079400 TI - Amyloid-beta peptide is toxic to neurons in vivo via indirect mechanisms. AB - We have studied the neurotoxicity of amyloid-beta (Abeta) after a single unilateral intravitreal injection. Within the retina apoptotic cells were seen throughout the photoreceptor layer and the inner nuclear layer but not in the ganglion cell layer at 48 h after injection of Abeta(1-42) compared to vehicle control and control peptide. At 5 months, there was a significant reduction in total cell numbers in the ganglion cell layer in Nissl stained retinas. There was glial cell dysfunction with upregulation of glial fibrillary acidic protein and a reduction in the expression of Muller cell associated proteins in the injected retinas. These results suggest an indirect cytotoxic effect of Abeta on retinal neurons and an important role for dysfunction of Muller glia in mediating Abeta neurotoxicity. PMID- 12079401 TI - Increased expression and redistribution of the antiapoptotic molecule Bcl-xL in Parkinson's disease. AB - In the present study, we tried to clarify the potentially protective role of Bcl x(L), an anti-apoptotic member of the Bcl-2 family of proteins, in Parkinson's disease (PD). Using in situ hybridization on human postmortem mesencephalon sections, we show that in PD patients Bcl-x(L) mRNA expression per dopaminergic neuron was almost double that of controls. We also show that, ultrastructurally, this effect may be mediated by a redistribution of Bcl-x(L) from the cytosol to the outer mitochondrial membrane. PMID- 12079402 TI - Dopamine has a critical role in photoreceptor degeneration in the rd mouse. AB - Photoreceptors receive paracrine input from dopaminergic interplexiform cells. Rod photoreceptors in the rd mouse degenerate rapidly due to a specific gene defect. We investigated the effects of dopamine on rd mouse photoreceptors in retinal organ culture. Retinas were harvested from rd or wild-type mice at postnatal day 2 and grown in organ culture for 27 days. When antagonists for either D(1)- or D(2)-family dopamine receptors were added to the media, photoreceptor degeneration was blocked. Furthermore, when dopamine was depleted by the addition of 6-hydroxydopamine and pargyline, photoreceptor survival appeared comparable to wild-type retinal cultures. The addition of a dopamine agonist induced photoreceptor degeneration in dopamine-depleted rd organ cultures. In all cases, photoreceptors maintained robust staining of opsin. These results demonstrate that dopamine antagonists or dopamine depletion blocks photoreceptor degeneration and that dopamine is necessary for photoreceptor degeneration in the rd mouse retinal organ culture model, indicating that dopamine antagonists may represent a therapeutic strategy in retinal degenerative disease. PMID- 12079403 TI - Depolarization-induced 65zinc influx into cultured cortical neurons. AB - Toxic Zn(2+) influx may be a key mechanism underlying selective neuronal death after transient global ischemia in rats. To identify routes responsible for neuronal Zn(2+) influx, we measured the accumulation of (65)Zn(2+) into cultured murine cortical cells under depolarizing conditions (60 mM K(+)) associated with severe hypoxia-ischemia in brain tissue. Addition of 60 mM K(+) or 300 microM kainate substantially increased (65)Zn(2+) accumulation into mixed cultures of neurons and glia, but not glia alone. (65)Zn(2+) accumulation was attenuated by increasing concentrations of extracellular Ca(2+) or trypsin pretreatment, but not by late trypsinization, and corresponded to an increase in atomic Zn(2+). Confirming predominantly neuronal entry, K(+)-induced (65)Zn(2+) accumulation was reduced by prior selective destruction of neurons with NMDA. K(+)-induced (65)Zn(2+) influx was not sensitive to glutamate receptor antagonists, but was attenuated by Gd(3+) and Cd(2+) as well as 1 microM nimodipine; it was partially sensitive to 1 microM omega-conotoxin-GVIA, and insensitive to 1 microM omega agatoxin-IVA. K(+)-induced, Gd(3+)-sensitive (45)Ca(2+) accumulation but not (65)Zn(2+) accumulation was sharply attenuated by lowering extracellular pH to 6.6. PMID- 12079404 TI - Dystrophin stabilizes alpha 3- but not alpha 7-containing nicotinic acetylcholine receptor subtypes at the postsynaptic apparatus in the mouse superior cervical ganglion. AB - The nicotinic acetylcholine receptor (nAChR) subtypes were characterized in the superior cervical ganglion (SCG) of wild-type and dystrophin-lacking mdx mice. The binding of Epibatidine and alphaBungarotoxin, ligands for alpha3- and alpha7 containing receptors, respectively, revealed, for each ligand, a single class of high-affinity binding sites, with similar affinity in both wild-type and mdx mice. The Epibatidine-labeled receptors were immunoprecipitated by antibodies against the alpha3, beta2, and beta4 subunits. Immunocytochemistry showed that the percentage of alpha3-, beta2-, and beta4- but not of alpha7-immunopositive postsynaptic specializations was significantly lower in mdx than in wild-type mouse SCG. These observations suggest that the mouse SCG contains nAChRs, stabilized by dystrophin, in which the alpha3 subunit is associated with the beta2 and/or beta4 subunits. Conversely, dystrophin is not involved in the stabilization of the alpha7-containing nAChRs, as the percentage of alpha7 immunopositive synapses is similar in both wild-type and mdx mouse SCG. PMID- 12079406 TI - Cloning, expression and characterization of the CHO cell elongating factor (Cef) from Vibrio cholerae O1. AB - CHO cell-elongating factor (Cef) is a recently identified putative virulence factor of Vibrio cholerae. Our previous studies show that this 85 kDa protein elongates CHO cells, causes fluid accumulation in suckling mice and has esterase activity. In this study, the cef gene was cloned in Escherichia coli using a yeast vector and subsequently expressed in the yeast Pichia pastoris. The cef genes from V. cholerae candidate vaccine strains JBK 70 and CVD 103-HgR were sequenced and found to be nearly identical (100 and 99.9% respectively) with an open reading frame (ORF) from the published sequence of V. cholerae N16961. Cloned toxin was purified to homogeneity in 3 steps using anion exchange, hydrophobic interaction and gel filtration chromatography. The size of cloned Cef on SDS-PAGE gels was 114 kDa. The increased size was probably due to glycosylation by the yeast since cloned protein reacted strongly with a glycoprotein stain. The cloned protein could not be directly sequenced, but when treated with trypsin, yielded a protein fragment with an amino acid sequence that matched the sequence predicted for the Cef protein. The purified cloned protein had esterase and CHO cell activity, but no suckling mouse activity. PMID- 12079405 TI - Early cell signaling by the cytotoxic enterotoxin of Aeromonas hydrophila in macrophages. AB - A cytotoxic enterotoxin (Act) of Aeromonas hydrophila is an important virulence factor with hemolytic, cytotoxic and enterotoxic activities. In this report, we demonstrated Act rapidly mobilized calcium from intracellular stores and evoked influx of calcium from the extracellular milieu in macrophages. A direct role of calcium in Act-induced prostaglandin (e.g. PGE(2)) and tumor necrosis factor alpha (TNF alpha) production was demonstrated in macrophages using a cell permeable calcium chelator BAPTA-AM, which also down-regulated activation of transcription factor NF-kappa B. We showed that Act's capacity to increase PGE(2) and TNF alpha production could be blocked by inhibitors of tyrosine kinases and protein kinase A. In addition, Act caused up-regulation of the DNA repair enzyme redox factor-1 (Ref-1), which potentially could promote DNA binding of the transcription factors allowing modulation of various genes involved in the inflammatory response. Taken together, a link between Act-induced calcium release, regulation of downstream kinase cascades and Ref-1, and activation of NF kappa B leading to PGE(2) and TNF alpha production was established. Since Act also caused extensive tissue damage, we showed that Act increased reactive oxygen species, and the antioxidant N-acetyl cysteine, blocked Act-induced PGE(2) and TNF alpha production, as well as NF-kappa B nuclear translocation in macrophages. We have demonstrated for the first time early cell signaling initiated in eukaryotic cells by Act, which leads to various biological effects associated with this toxin. PMID- 12079407 TI - Proteolysis of interleukin-6 receptor (IL-6R) by Porphyromonas gingivalis cysteine proteinases (gingipains) inhibits interleukin-6-mediated cell activation. AB - Current consensus is that periodontitis is an infectious disease in which a deregulated chronic inflammatory reaction not only may lead to periodontal tissue damage but also eventually may cause tooth loss. In controlling the inflammatory state the interplay between a network of cytokines and their receptors plays an important role. Here we show that the interleukin-6 receptor (IL-6R) is rapidly and efficiently inactivated by gingipains, the arginine- (HRgpA and RgpB) and lysine- (Kgp) specific cysteine proteinases from Porphyromonas gingivalis. Preincubation of HepG2 cells with active gingipains results in the loss of gp80 (CD126) from the cell surface. This also correlates with a decreased responsiveness to stimulation by interleukin-6 (IL-6), as determined by measurement of the status of IL-6R-mediated STAT 3 (Signal Transducer and Activator of Transcription 3) activation by this cytokine. Significantly, incubation of cells with gingipains was not accompanied by release of the soluble receptor, indicating its degradation, and this was confirmed by susceptibility of the recombinant, soluble receptor to proteolytic digestion by these enzymes. With the exception of the degradation of soluble IL-6R (sIL-6R) by Kgp, all of these reactions were also observed in the presence of serum suggesting that receptor inactivation may occur in vivo. Interestingly, Kgp, although less effective in cleaving sIL-6R, was able to decrease cell responsiveness to IL-6, possibly through degradation/inactivation of the signal transducing component (gp130) associated with IL-6R. These data, together with previous observation that IL-6 itself is inactivated by gingipains, suggest that at periodontitis sites infected by P. gingivalis the inflammatory reactions dependent on IL-6 could be severely hindered contributing to both tissue damage and periodontopathogen survival. PMID- 12079408 TI - Silkworm larvae as an animal model of bacterial infection pathogenic to humans. AB - Silkworm larvae, Bombyx mori, were examined as an animal model of human infection with pathogenic bacteria. When 3 x 10(7) cells of Staphylococcus aureus (S. aureus), Pseudomonas aeruginosa, or Vibrio cholerae were injected into the blood of fifth instar silkworm larvae, over 90% of the larvae died within 2 days, whereas over 90% survived for 5 days after injection of the same amount of Escherichia coli. Growth of S. aureus was observed in larvae blood and tissues. Immunostaining analysis revealed that S. aureus proliferated at the surface of the midgut. Infection of silkworm larvae by methicillin-sensitive S. aureus was cured by ampicillin, oxacillin, and vancomycin, whereas infection by methicillin resistant S. aureus was not cured by ampicillin or oxacillin, although vancomycin was effective. Disinfectants were not effective because of toxicity against the larvae. Thus, silkworm larvae are useful for evaluating antibiotics for pathogenic bacterial infection in humans. PMID- 12079409 TI - Streptococcus anginosus adheres to vascular endothelium basement membrane and purified extracellular matrix proteins. AB - The mechanisms of bacterial adherence in the initial stages of native valve endocarditis are unclear, especially in patients without valve disease or the presence of a platelet-fibrin thrombus. Extracellular matrix may act as a ligand in areas of exposed basement membrane on the endothelial monolayer. In this study, adherence of 55 clinical blood and 21 oral viridans streptococcal isolates was examined using purified extracellular matrix compounds. The majority of blood and oral isolates exhibited adherence to purified laminin, fibronectin, and fibrinogen, with lesser adherence to type I and IV collagens. Adherence to laminin and fibronectin was concentration dependent, saturable, and competitively inhibited with soluble ligand. A Streptococcus anginosus isolate and other viridans strains exhibiting a strong laminin adherence phenotype bound extensively to the endothelial aspect of human and porcine valve tissue sections and were inhibited by soluble laminin and anti-laminin antibody fragments. Using a novel native porcine valve explant adherence model, we localized binding to areas of exposed basement membrane by confocal and scanning electron microscopy. These studies support the hypothesis that bacterial adherence to exposed basement membrane plays a role in the initial phase of native valve endocarditis. PMID- 12079411 TI - Latex allergy: historical perspective. AB - The widespread use of powdered latex gloves, following the institution of universal precautions in 1987, increased the occurrence of a host of adverse reactions to natural rubber latex (NRL). Although the most common reactions are irritant or allergic dermatitis reactions to manufacturing chemicals, a spectrum of type I IgE-mediated reactions to NRL proteins are of great concern. IgE mediated reactions range from contact urticaria to occupational asthma and anaphylaxis, resulting in significant morbidity and potential mortality. At-risk populations include highly exposed groups like health care workers and certain patient groups requiring extensive medical treatment. A significant complicating factor is the association with clinically significant adverse reactions to certain foods because of allergen cross-reactivity. Institution of policies to use only low-protein, powder-free gloves, has resulted in a dramatic reduction of visits to occupational health departments and workmen's compensation claims due to occupational asthma. PMID- 12079412 TI - Latex allergy: prevalence, risk factors, and cross-reactivity. AB - There are few little exact epidemiological data on the prevalence and incidence of latex allergy, partly because the diagnostic tools are unsatisfactory and partly because the epidemiological study planning often does not fulfill criteria of good praxis. On the basis of present data, latex allergy in normal population is low, under 1%. Known risk groups such as health care workers, atopic subjects, people with hand dermatitis, and especially spina bifida patients show higher prevalence numbers. The common serological cross-reactivity between latex and a great number of different fruits and vegetables is bound to common plant pathogenesis-related proteins and storage proteins. Despite positive serological tests, only about half of NRL-allergic subjects have clinical symptoms after eating cross-reacting foods. PMID- 12079413 TI - Natural rubber latex allergy in the occupational setting. AB - Over the last decade, the prevalence of natural rubber latex (NRL) allergy has reached epidemic proportions among workers who use or who are exposed to powdered latex products. NRL-associated occupational asthma is confined largely to those exposed to powdered latex glove use or other latex aerosols. The most frequent presenting symptom of NRL allergy is contact urticaria; inhalation may cause symptoms of allergic rhinitis and asthma. Skin prick testing is the most accurate tool for diagnosis of NRL allergy. The cornerstone of management is cessation of exposure; substitution with non-NRL or nonpowdered NRL gloves results in predictable rapid disappearance of latex aeroallergen. PMID- 12079414 TI - Diagnosis of natural rubber latex allergy. AB - The accurate diagnosis of a latex-allergic individual begins with a comprehensive clinical history. Atopy, food allergies, hand dermatitis, and temporal relationships between allergic symptoms and natural rubber product exposure are risk factors that increase the suspicion of latex allergy. If symptoms are temporally delayed (hours-days) and confined to skin-latex product contact areas, Type IV hypersensitivity should be suspected and patch testing may be performed to identify activated T cells that are specific for selected rubber chemical additives. If ocular, upper and lower airway, and/or systemic allergic symptoms are observed with rapid onset (minutes) following a definable latex exposure, Type I hypersensitivity should be suspected. One or several confirmatory tests for latex-specific IgE antibody in the skin or blood may next be performed to verify a sensitized (IgE antibody positive) state. If the clinical history remains discordant with a skin test or blood test result, in vivo provocation tests may be cautiously considered for adjudication. Diagnostic methods for latex specific IgE antibody detection in skin and blood are overviewed, with a focus on their performance, advantages, and limitations. PMID- 12079415 TI - Allergenic proteins of natural rubber latex. AB - As the living cytoplasm of laticiferous cells, Hevea brasiliensis latex is a rich blend of organic substances that include a melange of proteins. A small number of these proteins have given rise to the problem of latex allergy. The salient characteristics of H. brasiliensis latex allergens that are recognized by the International Union of Immunological Societies (IUIS) are reviewed. These are the proteins associated with the rubber particles, the cytosolic C-serum proteins and the B-serum proteins that originate mainly from the lutoids. Procedures for the isolation and purification of latex allergens are discussed, from latex collection in the field to various preparative approaches adopted in the laboratory. As interest in recombinant latex allergens increases, there is a need to validate recombinant proteins to ascertain equivalence with their native counterparts when used in immunological studies, diagnostics, and immunotherapy. PMID- 12079416 TI - Measurement of latex proteins and assessment of latex protein exposure. AB - Reduction of protein levels in manufactured natural rubber latex products is important for preventing sensitization and adverse allergic reactions to latex. Because of the complex nature of latex extracts, accurate protein measurement is a challenge. Standard total protein assays were effective in reducing protein levels from what were once extremely high levels, but these assays are plagued with false-positive reactions and limited sensitivity. An ELISA for antigenic protein has been standardized and promises to provide more consistent measurement of the proteins with potential to cause adverse reactions. Antigenic proteins represent the total protein fraction with potential to be allergenic. Measuring antigenic protein in a consistent manner should help to further reduce the level of sensitizing protein and further reduce allergic reactions to latex-medical products. PMID- 12079417 TI - Quantitation of latex allergens. AB - Minimizing allergen concentration in latex goods to prevent sensitization to natural rubber latex (NRL) and thereby the development of clinical allergy is acknowledged as of mutual interest for rubber manufacturers and regulatory health authorities. However, measuring total protein, the principal currently available method, cannot be deemed a satisfactory regulatory measure to control allergen content. Specific methods based on human IgE-containing reagents, such as radioallergosorbent test (RAST) inhibition, have been available in certain laboratories for demonstrating NRL allergens in rubber products but the methods lack standardization. Currently, one commercial test has become available for measuring individual NRL allergens by capture ELISA-based assays using monoclonal antibodies and purified or recombinant allergens. Such methods are specific, they can be properly standardized, and they are of sufficient sensitivity and reproducibility. Results from medical gloves collected in two national market surveys in Finland in 1995 and 1999, respectively, show that Hev b 6.02 and Hev b 5, the two major allergens for NRL-allergic adults, are the most abundant allergens regularly detectable in high- and moderate-allergen gloves. In addition, Hev b 3 and Hev b 1, the two major allergens for children with spina bifida, are also commonly found. In general, when the sum of the four allergens exceeded 1 microg/g, most NRL-allergic patients showed positive skin prick test reactions against them. Using these new methods assessment of threshold levels that could in due course become guidelines for the rubber industry and regulatory health authorities is becoming possible. Eventually, this progress is expected to lead to a declining incidence of latex allergy. PMID- 12079418 TI - Measurement of airborne latex allergens. AB - A method for the standardized sampling and quantification of aeroallergens was developed. It proved suitable for the analysis of allergen loads in a variety of medical settings. Several studies demonstrate that the use of powdered allergenic latex gloves leads to a significant aeroallergen load which is responsible for IgE-mediated rhinitis, bronchial asthma, and conjunctivitis in health care workers. Only nonpowdered latex gloves with low or no allergen content should therefore be used. PMID- 12079419 TI - Murine models for natural rubber latex allergy assessment. AB - Murine models provide a powerful tool in the investigation of latex allergy and the development of intervention strategies. The immune responses to protein allergens of mice and humans are similar but differences related to the roles of IgE and IgG must be recognized. Mice have been shown to mount a dose and time dependent IgE response to latex proteins following topical, respiratory, and subcutaneous exposures. Methods are available to evaluate cutaneous and respiratory responses to latex challenge in sensitized animals. These models have been used to investigate the role of route of exposure on the development of latex allergy and to provide a means for investigating the contribution of individual proteins to adverse respiratory and dermal responses. These models provide a mechanism for the evaluation of new technologies aimed at reducing the allergenicity of latex products, and for testing for the potential for cross reactivity to new allergens in previously sensitized individuals. Murine models may also provide a method for testing immunotherapy strategies prior to initiating human trials. PMID- 12079420 TI - Glove powder reduction and alternative approaches. AB - This article addresses the role of glove powder in facilitating allergic reactions to natural rubber latex (NRL) and to the chemical additives in synthetic and NRL gloves as well as its role in eliciting postsurgical complications. Various dusting powders have been used historically to prevent gloves from sticking to each other and to facilitate donning. All have manifested adverse consequences for health care professionals and patients. Manufacturing methods for powder reduction and elimination are presented. The recently developed ASTM methods for the quantitation of powder on powder-free and powdered gloves are reviewed along with the new ASTM maximum powder limits for all medical gloves. Caution must be exercised when methods of protein and powder reduction are implemented to minimize the possibility of creating other adverse consequences. PMID- 12079421 TI - Natural rubber latex protein reduction with an emphasis on enzyme treatment. AB - Natural rubber latex (NRL), derived from the Hevea brasiliensis tree, is a material used to manufacture products in health care, including medical gloves. Proteins are a naturally occurring component of NRL. These proteins, which can be present on the surface of NRL gloves, have been related to hypersensitivity reactions in some humans who come into contact with them. These same proteins also help to maintain the latex colloidal stability during collection and transport prior to manufacture. Consequently, when measures are taken to remove or degrade these proteins, other problems can be introduced, such as destabilization of the latex and changes in its coagulation properties. Practical methods are available to reduce the extractable antigenic protein content of NRL products. We describe here methods of reducing proteins in commercial-grade NRL and finished products. NRL gloves manufactured with adequate leaching can produce products with lower levels of extractable antigenic proteins. Emphasis is given here to enzyme treatment of NRL, as this process is very effective in reducing antigenic proteins in NRL. While this technology adds marginally to the production cost of standard grades of NRL, it is still quite cost-effective when compared with postwashing NRL products or the use of synthetic latex. Moreover, enzyme-treated NRL maintains the excellent physical properties and performance of NRL. PMID- 12079422 TI - Regulatory initiatives for natural latex allergy: US perspectives. AB - The US Food and Drug Administration (FDA) has regulatory authority over foods, human drugs, cosmetics, medical devices, radiological products, biologics, and veterinary products. Among these products, FDA believes that the use of medical devices, including medical gloves, condoms, catheters, and breathing bags, represents the greatest source of natural latex proteins to exposed individuals. A medical device is defined in the Federal Food Drug and Cosmetic Act (FFDCA) as an instrument, apparatus, implement, machine, etc., that is intended for use in the diagnosis or treatment of disease or is intended to affect the structure or any function of the body of a human or other animal, and that does not achieve any of its principal intended purposes through chemical action in the body. This article provides some brief, general background about FDA's medical device regulatory process and then addresses the issue of natural latex allergy. Finally we discuss the steps the Agency has taken to evaluate the magnitude and nature of the problem, and FDA's efforts to assist manufacturers, health professionals, and others in minimizing exposure and sensitization to natural latex proteins in medical devices. PMID- 12079423 TI - Medical devices manufactured from latex: European regulatory initiatives. AB - In Europe the marketing of medical devices manufactured from latex is regulated by directives describing the essential (safety) requirements that products have to fulfill to obtain marketing approval. This paper describes the general requirements for marketing medical devices in Europe and, more specifically, the requirements for products manufactured from natural rubber latex. The requirements for marketing medical devices can be fulfilled by using the relevant harmonized European standards. These standards are regularly under revision to incorporate the latest scientific developments. For certain devices, for example, latex medical (examination and surgical) gloves, specific standards have been published. Medical devices manufactured from latex pose a serious problem because of the risk of induction of allergy both against the latex proteins inherently present (type I or immediate type allergy) and against chemicals added during processing (type IV or delayed type hypersensitivity) present as residues in the latex products. So, besides requirements for product quality in terms of barrier properties, strength, and sterility, the main focus consists of the allergy inducing properties of the latex products. Recent developments have reopened the discussion on the value of total protein versus allergen determination in latex medical gloves. However, as long as minimal levels needed for both sensitization and elicitation have not been established, a safe maximum level for leachable proteins/allergens in latex products cannot be determined. A European Commission guidance document on the latex allergy problem is currently being drafted by experts from Competent Authorities. PMID- 12079424 TI - A physiologically based pharmacokinetic model of p,p'-dichlorodiphenylsulfone. AB - A physiologically based pharmacokinetic model of the absorption, distribution, metabolism, and elimination of p,p'-dichlorodiphenylsulfone (DDS) in male and female rats and mice is presented. Data used in constructing the model come from single-dose intravenous administration of DDS to male Fischer 344 rats (10 mg/kg, with data taken up to 504 h after administration), from single-dose gavage administration to male rats (10, 100, or 1000 mg/kg, with data up to 72 h after administration), and from chronic feed studies in male and female rats and male and female B6C3F(1) mice (studies of duration from 2 weeks up to 18 months, with feed concentrations of DDS up to 300 ppm). The model uses diffusion-limited kinetic for the distribution of the parent compound. Because fewer data are available for the metabolites of DDS (at least five of which are known to exist in the data), the model groups the metabolites into one metabolic pathway and uses simpler flow-limited kinetics for the metabolites. The data show that the kinetics of DDS are nonlinear. Possible sources of nonlinearity considered in the model were nonlinear (Michaelis-Menten) metabolism, nonlinear absorption of DDS from the gut, and induction by DDS of its own metabolism. A model using Michaelis Menten metabolism was not found to give a significantly better fit than one using first-order linear metabolism, but omitting either of the other nonlinear effects was found to give a significantly poorer fit to the data. Because the data from mice are limited compared to those from rats, there is more confidence in the model's description of DDS kinetics in rats than in its description of kinetics in mice. PMID- 12079425 TI - Pyridostigmine bromide modulates the dermal disposition of [14C]permethrin. AB - The cause of the Gulf War Syndrome may be related to soldiers being exposed to insecticides (e.g., permethrin (P)), insect repellents (e.g., N,N-diethyl-m toluamide (DEET)), an organophosphate nerve agent simulant (e.g., diisopropyl fluorpohosphate (DFP)), and/or prophylactic treatment (e.g., pyridostigmine bromide (PB)) against potential nerve gas attacks. The purpose of this study was to assess the dermal disposition of [14C]permethrin in ethanol or ethanol:water (3:2) in the isolated perfused porcine skin flap (IPPSF) model with simultaneous dermal exposure to DEET or DFP. These IPPSFs were also simultaneously perfused arterially with or without PB, DFP, or DFP + PB. The results indicated that DFP + PB significantly increased [14C]permethrin absorption compared to controls (1.06% dose vs 0.14% dose). PB significantly increased [14C]permethrin disposition in the stratum corneum (SC) in aqueous mixtures only (9.40 vs 3.35% dose), while topical DEET or topical DFP reduced [14C]permethrin levels in the SC especially in nonaqueous mixtures. PB also significantly enhanced [14C]permethrin penetration into all skin tissues and perfusate in aqueous mixtures, while DEET reversed this effect. PB appeared to influence [14C]permethrin disposition in flowthrough diffusion cells, suggesting that the mechanism of this interaction may be associated predominantly with epidermal permeability, although muscarinic effects in the vasculature in IPPSFs should not be ruled out and requires further investigation. These experiments suggest that intraarterial perfusion of PB and/or DFP and topical application of DFP or DEET can alter the disposition of [14C]permethrin in skin and possibly its bioavailability in soldiers simultaneously exposed to these chemicals. PMID- 12079427 TI - Identification of PLA(2) and alpha-neurotoxin proteins in the venom of Pseudonaja affinis (dugite). AB - The Western brown snake Pseudonaja affinis (dugite), common to the Perth area of Western Australia, possesses one of the most lethal venoms in the world. Little is known, however, about the toxic protein constituents of the venom, other than those causing coagulopathic and procoagulant effects. The current study was therefore undertaken in order to identify other protein constituents and activities present. Crude venom induced a contraction in rat tracheal preparations through phospholipase A(2) (PLA(2)) activity, as shown by the complete and partial inhibition of contraction by PLA(2) inhibitors 4 bromophenacyl bromide and quinacrine. Further, a reduced degree of smooth muscle contraction in the presence of the leukotriene receptor antagonist SKF104353 suggested that this effect was mediated by leukotriene metabolites. The venom induced contraction did not reoccur upon a second administration of the venom, despite the muscle retaining its contractile function and appearing histologically undamaged. Chromatographic separation of the protein constituents of the venom showed that PLA(2) activity was associated with all protein fractions. A low-molecular-weight component of the venom was further investigated through N-terminal sequencing and found to possess high identity to the short chain alpha-neurotoxin family of toxins. Venom activity on cultured rat cardiac myocytes and cultured cortical neurons was also examined. The crude venom was found to temporarily inhibit the beating of the cardiac myocytes, after which the beating resumed erratically. Cortical neurons, however, were irreversibly affected, showing concentration-dependent cell death. PMID- 12079426 TI - Proinflammatory properties of coplanar PCBs: in vitro and in vivo evidence. AB - So-called coplanar polychlorinated biphenyls (PCBs), as well as other environmental contaminants that are aryl hydrocarbon receptor (AhR) agonists, may compromise the normal functions of vascular endothelial cells by activating oxidative stress-sensitive signaling pathways and subsequent proinflammatory events critical in the pathology of atherosclerosis and cardiovascular disease. To test this hypothesis, porcine endothelial cells were exposed to PCB 153 and to three coplanar PCBs (PCB 77, PCB 126, or PCB 169). In contrast to PCB 153, which is not a ligand for the Ah receptor (AhR), all coplanar PCBs disrupted endothelial barrier function. All coplanar PCBs increased expression of the CYP1A1 gene, oxidative stress (DCF fluorescence), and the DNA-binding activity of nuclear factor kappaB (NF-kappaB). PCB-induced oxidative stress was concentration dependent, with PCB 126 exhibiting a maximal response at the lowest concentration (0.5 microM) tested. The increase in NF-kappaB-dependent transcriptional activity was confirmed in endothelial cells by a luciferase reporter gene assay. In contrast to PCB 153, coplanar PCBs that are AhR ligands increased endothelial production of interleukin-6. At 3.4 microM, expression of the adhesion molecule VCAM-1 was most sensitive to PCB 77 and 169. We also provide in vivo evidence, suggesting that binding to the AhR is critical for the proinflammatory properties of PCBs. Twenty hours after a single administration of PCB 77, VCAM-1 expression was increased only in wild-type mice, while mice lacking the AhR gene showed no increased staining for VCAM-1. These data provide evidence that coplanar PCBs, agonists for the AhR, and inducers of cytochrome P450 1A1, produce oxidative stress and an inflammatory response in vascular endothelial cells. An intact AhR may be necessary for the observed PCB-induced responses. These findings suggest that activation of the AhR can be an underlying mechanism of atherosclerosis mediated by certain environmental contaminants. PMID- 12079428 TI - Diol- and triol-type ginseng saponins potentiate the apoptosis of NIH3T3 cells exposed to methyl methanesulfonate. AB - In this study we investigated the effect of ginseng saponins on the p53-dependent apoptosis in NIH3T3 cells exposed to methyl methanesulfonate (MMS), an alkylating agent. Trypan blue exclusion assay, cell morphology studies, and apoptotic index determined by acridine orange staining showed that the postincubation of MMS exposed cells in medium containing diol- (PD) or triol-type (PT) ginseng saponins potentiate the apoptotic cell death. FACS analysis indicated that the increased apoptotic cell population in the saponin-postincubation group was accompanied by the accumulation of cells in G0/G1 phase. By Western blot analyses it was demonstrated that postincubation of saponins increases the expression of p53 and p21 in MMS-exposed cells but decreased that of CDK2, cyclin E and D1, and PCNA. The upregulation of p53 and p21 and downregulation of CDK2 was shown to be p53 dependent in experiments using the p53 antisense oligonucleotide. These results suggest that ginseng saponins contain components potentiating the apoptosis of MMS-exposed NIH3T3 cells via p53 and p21 activation, accompanied with by downregulation of cell cycle-related protein expression. PMID- 12079429 TI - Induced proliferation of human MRC-5 cells by nitrogen oxides via direct and indirect activation of MEKK1, JNK, and p38 signals. AB - Nitrogen oxides (NOx) are important indoor air pollutants and an occupational hazard. Many studies demonstrated that NOx causes lung tissue damage based on the oxidation properties and the free-radical potentials of these gases. In this study we found that NOx delivered as a NO gas-saturated solution induced proliferation of human lung fibroblast MCR-5 cells as evidenced by increasing cell number and S phase distribution. Western data showed that NOx increased the expressions of c-Fos, c-Jun, and signaling kinases including MEKK1, JNK1, and p38 (with induction fold of 3.3, 2.8, and 3.2, respectively) in the cells 12 h after treatment. The levels of phospho-MEKK1 and phospho-JNK1 were also increased. The application of iNOS inhibitor, NAME, partially blocked the activation of MEKK4 and JNK1. These data suggested that JNK and p38 signaling kinases are activated partly by endogenous NO that are generated from NOx-activated iNOS in MRC-5 cells. Therefore, the NOx-induced cell proliferation via activation of MEKK1, JNK1, and p38 might contribute to lung tissue damage caused by NOx pollutants. PMID- 12079430 TI - The mycotoxin patulin alters the barrier function of the intestinal epithelium: mechanism of action of the toxin and protective effects of glutathione. AB - Patulin is a mycotoxin mainly found in apple and apple products. In addition to being toxic for animals, mutagenic, carcinogenic and teratogenic, patulin induces intestinal injuries, including epithelial cell degeneration, inflammation, ulceration, and hemorrhages. In a study of the cellular mechanisms associated with the intestinal toxicity of patulin, two human epithelial intestinal cell lines (HT-29-D4 and Caco-2-14) were exposed to the mycotoxin. Micromolar concentrations of patulin were found to induce a rapid and dramatic decrease of transepithelial resistance (TER) in both cell lines without major signs of toxicity as assessed by the LDH release assay. Since TER reflects the organization of tight junctions, these data indicate that patulin affected the barrier function of the intestinal epithelium. The inhibitory effect of patulin on TER was closely associated with its reactivity for SH groups: (i) cysteine and glutathione prevented the cells from patulin injury; (ii) patulin toxicity was potentiated by buthionine sulfoximine, a specific glutathione-depleting agent; (iii) treatment of the cells with N-ethylmaleimide, a compound known to react with SH groups, resulted in a marked decrease of TER. Moreover, the inhibitory effect of patulin on TER was mimicked and potentiated by phenylarsine oxide, a specific inhibitor of protein tyrosine phosphatase (PTP). This cellular enzyme is a key regulator of intestinal epithelial barrier function. The active site of PTP contains a cysteine residue (Cys215) that is essential for phosphatase activity. Sulfhydryl-reacting compounds such as acetaldehyde decrease TER through covalent modification of Cys215 of PTP. We propose that the toxicity of patulin for intestinal cells involves, among other potential mechanisms, an inactivation of the active site of PTP. PMID- 12079431 TI - Genistein modulates splenic natural killer cell activity, antibody-forming cell response, and phenotypic marker expression in F(0) and F(1) generations of Sprague-Dawley rats. AB - The potential effects of the phytoestrogen genistein (GEN) on the immune system were evaluated in both F(0) (dams) and F(1) generations of Sprague-Dawley rats exposed to a soy-free diet containing low (L: 25 ppm), middle (M: 250 ppm), and high (H: 1250 ppm) levels of GEN. In dams, exposure to GEN from Gestation Day 7 to Postpartum Day 51 (totally 65 days) produced a significant increase in NK cell activity (M and H), while a decrease in the percentage of helper T cells (H). In F(1) males, exposure to GEN gestationally, lactationally, and through feed from Postnatal Days 22 to 64 (total 78 days) produced an increase in the relative weights (% body) of spleen (L and H) and thymus (L). Furthermore, exposure to GEN increased the number of splenic B cells (H), T cells (L, M, and H), and T-cell subsets (L, M, and H). Although GEN decreased the percentages of splenic NK cells (L, M, and H), no effect on the activity of NK cells was observed. In F(1) females, exposure to GEN produced a decrease in terminal body weight (H), with an increase in the relative weight of spleen (L, M, and H). Exposure to GEN also increased the number of splenic B cells (L), macrophages (L and M), T cells (H), helper T cells (L and H), and cytotoxic T cells (M and H). Additionally, exposure to GEN increased the percentages of T cells (M and H), helper T cells (H), and cytotoxic T cells (M and H). Moreover, the spleen IgM antibody-forming cell response to sheep red blood cells was enhanced (H), although the percentages of B cells were decreased (M and H). No effect on the activity of NK cells was observed; however, the percentages of splenic NK cells were decreased by GEN (L and H). In conclusion, these results demonstrate that exposure to GEN can modulate the immune responses in Sprague-Dawley rats. Furthermore, the sexual dimorphic effects of GEN in F(1) male and female rats suggest that there may be interactions between GEN and the responses modulated by sex hormones. PMID- 12079432 TI - Hypochlorous acid is a potent inhibitor of acetylcholinesterase. AB - The role of hydrogen peroxide and peroxynitrite in the induction of airway hyperreactivity has been well described. Another reactive species which is formed during airway inflammation is hypochlorous acid (HOCl). In the present investigation the effect of HOCl on cholinergic innervation of the airway was investigated. It was observed that HOCl was capable of increasing the basal tension of electrically stimulated tracheal smooth muscle. It was found that HOCl inhibits purified acetylcholinesterase with an IC50 value of 0.66 microM. Decreased acetylcholinesterase activity could allow accumulation of acetylcholine and increased airway muscle tension. The effects of HOCl on the isolated organ and the enzyme preparation could be precluded with thiol group-containing compounds such as reduced glutathione and N-acetylcysteine. The present findings indicate that HOCl can act as inhibitor of acetylcholinesterase. The implications of this finding for the induction of airway hyperreactivity are discussed. PMID- 12079434 TI - Obesity and associated coronary heart disease risk factors in a population of low income African-American and white women: the North Carolina WISEWOMAN project. AB - BACKGROUND: Obesity has been associated with many co-occurring coronary heart disease (CHD) risk factors as well as CHD mortality. These associations have been shown to vary between African-American and white sample populations. METHODS: The authors examined whether obesity co-occurs with several CHD risk factors (diabetes, hypertension, hypercholesterolemia, low high-density lipoprotein cholesterol (HDL-C)), and estimated the 10-year risk for CHD in the North Carolina WISEWOMAN (Well Integrated Screening and Evaluation for Women Across the Nation) study sample. This sample includes low-income African-American and white women (> or = 50 years of age). RESULTS: Among white women (n = 1,284), 34% were overweight (BMI = 25.0-29.99 kg/m(2)) and 35% obese (BMI > or = 30 kg/m(2)); among African-American women (n = 754), 28% were overweight and 59% obese. Among obese and nonobese African-American women, the prevalence of three or more co occurring risk factors was similar (obese = 17.7% (95% confidence interval (CI): 13.9, 21.6) and nonobese = 13.3% (95% CI: 8.7, 17.8)). By contrast, the prevalence among white women was greater among the obese (26.9% (95% CI: 22.9, 31.0)) than the nonobese (13.0% (95% CI: 9.7, 16.2)). CONCLUSIONS: The differences between and within African-American and white women may be accounted for by the high levels of HDL-C among obese and nonobese African-American women. PMID- 12079435 TI - Page for patients. Water safety. PMID- 12079436 TI - A healthy diet indicator: quantifying compliance with the dietary guidelines using the BRFSS. AB - BACKGROUND: The Behavioral Risk Factor Surveillance System (BRFSS) monitors population prevalence of risk factors related to chronic disease through annual telephone surveys. The purpose of this study was to investigate the feasibility of using the BRFSS to estimate an indicator of overall diet quality among adults. METHODS: The authors developed a brief set of dietary questions that were included in the 1997 Michigan BRFSS. Responses to these questions were analyzed to calculate the Michigan healthy diet indicator (MI-HDI), which was modeled after the U.S. Department of Agriculture's Healthy Eating Index. RESULTS: Among 2,532 respondents, the mean MI-HDI was 55.7 points (maximum score = 100). The mean MI-HDI decreased consistently with declining self-rated eating habits, from 61.5 among those with excellent eating habits to 45.2 among those with poor habits. Diet quality, as measured by the MI-HDI, was higher among women than men and improved with age, education, and factors related to certain interactions with health professionals. CONCLUSIONS: A BRFSS-based indicator can be a feasible and valuable tool for evaluating diet quality among adults using an established state-level surveillance system. PMID- 12079437 TI - Comparing the benefits of diet and exercise in the treatment of dyslipidemia. AB - BACKGROUND: Lifestyle changes are advocated as a first line of treatment for dyslipidemia. However, few studies have directly compared various combinations of diets and exercise. METHODS: In a randomized controlled pilot study, we compared the standard lifestyle recommendations (NCEP step I diet with regular exercise) and more intense interventions including the NCEP step I diet with a supervised aerobic exercise program and the step II diet with and without a supervised aerobic exercise program. We measured risk factors, dietary intake, time on treadmill, and health-related quality of life at baseline and after 3 months. RESULTS: Out of 198 eligible subjects, 47 (24%) were willing to participate and 41 completed the study. No significant change were observed with standard lifestyle recommendations. In contrast, participants in the more intense interventions lost weight (-1.7 to -3.7 kg) and reduced their total cholesterol ( 4% to -6%), low-density lipoprotein cholesterol (-6%), and systolic blood pressure (-7.3 to -8.8 mmHg). Participants in the exercise program significantly increased their exercise capacity (1.6 to 1.9 METS). Overall, each 10% reduction in body weight was associated with a 7.6% reduction in low-density lipoprotein cholesterol. CONCLUSION: Standard lifestyle recommendations had little effect on blood lipid levels but more intense lifestyle interventions may be effective at improving blood lipids, other risk factors, and quality of life. PMID- 12079438 TI - The efficacy of behavioral interventions to modify dietary fat and fruit and vegetable intake: a review of the evidence. AB - BACKGROUND: The evidence suggesting that nutrition, particularly dietary saturated fat and fruit and vegetable intake, is related to chronic disease risk has prompted considerable research on behavioral interventions focusing on dietary change. No clear understanding has emerged, however, of the degree to which these interventions can materially influence dietary change, or the types of intervention that are most effective and for whom. Therefore, the primary objective of the current study was to evaluate the overall effectiveness of behavioral dietary interventions in promoting dietary change related to chronic disease risk reduction. A secondary goal was to explore the relative effectiveness of specific intervention features and among different population subgroups. METHODS: We conducted an evidence-based review and secondary analysis of existing literature. Our data sources included reports of randomized controlled trials and other study designs identified from multiple searches of MEDLINE, EMBASE, PsycINFO, CINAHL, AGELINE, and AGRICOLA. We included all studies on humans (including children, adolescents, and adults) published in English since 1975 that had been conducted in North America, Europe, or Australia; that had sample sizes of at least 40 subjects at follow-up; that were not based on controlled diets; and that otherwise met inclusion criteria. Through dual review, we abstracted detailed information on study characteristics, methodology, and outcomes relating to consumption of fruits, vegetables, and fats. RESULTS: From 907 unduplicated articles originally identified, we retained 104 articles reporting on 92 independent studies. The studies were similarly successful in reducing intake of total and saturated fat, and increasing fruit and vegetable intake. More than three-quarters of the studies (17 of the 22 reporting results for fruit and vegetable intake) reported significant increases in fruit and vegetable intake, with an average increase of 0.6 servings per day. Similar consistent decreases were seen in intake of saturated fat and total fat (7.3% reduction in the percentage of calories from fat). Interventions appeared to be more successful at positively changing dietary behavior among populations at risk of (or diagnosed with) disease than among general, healthy populations. Two intervention components seemed to be particularly promising in modifying dietary behavior-goal setting and small groups. CONCLUSIONS: The majority of the interventions reviewed resulted in meaningful improvements in dietary factors behaviors associated with the prevention of chronic disease, particularly among individuals at elevated disease risk. The lack of similarity across studies in outcome measures, study design, analysis strategy, and intervention technique hampered our ability to draw broad conclusions about the most effective behavioral dietary interventions, but our findings offer insight into intervention components that may hold promise for future research efforts. PMID- 12079439 TI - Association between serum apolipoprotein C(II) concentration and coronary heart disease. AB - BACKGROUND: Apolipoprotein (Apo) C(II) plays an essential role in the metabolism of triglyceride-rich lipoproteins (TRLs) by activating lipoprotein lipase. Several studies have suggested that an abnormal concentration of Apo C(II) may serve as a marker for deficient TRL metabolism, a possible cause of coronary heart disease (CHD). The aim of the present study was to evaluate the association between Apo C(II) and the presence of CHD. METHODS: A case-control study comparing 352 CHD patients and 395 controls was performed. Demographic, clinical, and laboratory characteristics were determined. RESULTS: The CHD patients had a higher mean concentration of Apo C(II) than controls (5.3 mg/dl compared with 4.2 mg/dl, P < 0.001). Elevated serum Apo C(II) concentration was associated with CHD presence after adjustment for cardiovascular risk factors. The risk factor adjusted odds ratio (OR) for CHD was 1.60 (95% CI: 1.31-1.94) per 1 mg/dl increment in Apo C(II), compared with a risk factor-adjusted OR of 1.05 (95% CI: 0.85-1.32) per 40 mg/dl increment in triglyceride concentration. CONCLUSION: Increased serum concentration of Apo C(II) may represent a more sensitive marker of CHD than high serum concentration of triglycerides. Confirmation in cohort studies in required to establish or refute the role of elevated serum Apo C(II) as a risk factor for CHD. PMID- 12079440 TI - Lipid-lowering medication for secondary prevention of coronary heart disease in a German outpatient population: the gap between treatment guidelines and real life treatment patterns. AB - BACKGROUND: Few published data in particular from the United States indicate that the implementation of guidelines for prevention of coronary heart disease (CHD) is far from optimal. The objective of our study was to identify the type and prevalence of lipid-lowering medications in a German outpatient CHD population and to examine the impact of applied treatment regimens on serum lipid levels. METHODS: Retrospective analysis of the washout phase of 2,856 CHD patients requiring lipid-lowering medication. Data are derived from a multicenter, randomized, open-label, parallel group clinical trial comparing the safety and efficacy of atorvastatin versus simvastatin in 591 centers in Germany. Medical history, physical examination, and serum lipid levels were obtained at the beginning of the washout phase (Week -6) and at the end of the washout phase (Week -1, i.e., 5 weeks after the discontinuation of all prior lipid-lowering medications). The data at Week -6 represented the lipid levels under real life conditions. The difference from the data at Week -1 reflected the therapeutic effects achieved by the previous lipid-lowering treatment. RESULTS: The mean low density lipoprotein cholesterol (LDL-C) level at Week -6 was 173.4 +/- 42.5 mg/dl. Only 176 (6.2%) of 2,856 CHD patients were found to meet the target LDL-C level of <115 mg/dl at Week -6, only 76 (2.7%) patients had LDL-C levels <100 mg/dl, and 363 (12.7%) patients had LDL-C levels <130 mg/dl. After discontinuation of all prior lipid-lowering medications, mean LDL-C increased to 187.2 +/- 44.0 mg. This means that only a marginal 7.4% reduction in LDL-C level was achieved under real life treatment conditions. This limited LDL-C reduction was due mainly to the low prevalence of lipid-lowering treatment (65.5% of patients did not receive any medication at all) and inadequate dosing. With respect to the effect on LDL-C and total cholesterol, statins alone were superior to fibrates. CONCLUSION: The study shows that there is a wide gap between treatment guidelines and real life treatment patterns in Germany. Awareness of the risks of high cholesterol levels has to be increased among both patients and physicians. Available treatment guidelines should be better implemented. PMID- 12079441 TI - Development of questionnaires to measure physical activity cognitions among Taiwanese adolescents. AB - BACKGROUND: Although a number of studies have investigated perceived self efficacy, perceived benefits, and perceived barriers and their relationship to adolescents' participation in physical activity, the validity and reliability of the instruments used have not been consistently examined. The purpose of the present study was to develop culturally sensitive, reliable, and valid instruments to measure physical activity cognitions among Taiwanese adolescents. METHODS: The items generated for the instruments were drawn from a comprehensive literature review and a focus group interview. The instruments were translated using the back-translation technique and evaluated by a panel of experts. Psychometric testing was performed and confirmed with convenience samples of 110 and 969 Taiwanese adolescents. RESULTS: Psychometric testing demonstrated satisfactory internal consistency and validity of the instrument for this group of Taiwanese adolescents. Cronbach's alpha's ranged from 0.79 to 0.90. Construct validity was supported by exploring the factor structure of the instrument using confirmatory factor analysis and correlation with the outcome variable of physical activity. CONCLUSIONS: The study identified culturally sensitive items to measure physical activity cognitions in urban Taiwanese students in their early adolescence. Clinicians or practitioners can use the instruments developed and tested in this study for clinical assessment to identify risk factors for inactivity as well as for further research in similar populations. PMID- 12079442 TI - Three years after the dutch folic acid campaign: growing socioeconomic differences. AB - BACKGROUND: We investigated periconceptional awareness and the use of folic acid tablets 3 years after the 1995 campaign in the northern Netherlands. We studied whether the socioeconomic differences we found in 1996 with respect to awareness and use of folic acid remained in 1998. METHODS: The present study took place in 1998. Pregnant women were asked to complete a questionnaire. RESULTS: A total of 452 women completed the questionnaire in 1998. Sixty percent of the women with a lower level of education and 85% of the women with a higher level of education had heard of folic acid before pregnancy. Newspapers and magazines were the most often cited sources of information. Fifty percent of the women with a lower educational level used folic acid, while this figure was 80% for the group with higher education. CONCLUSION: Awareness and use of folic acid increased after the 1995 campaign. However, socioeconomic differences with respect to awareness of folic acid remained and even increased with respect to knowledge of the advised period. Use of folic acid is also associated with level of education more strongly than in 1996. Therefore, fortification of staple foods is an attractive option. PMID- 12079443 TI - Is Bacillus Calmette-Guerin revaccination necessary for Japanese children? AB - BACKGROUND: Bacillus Calmette-Guerin (BCG) revaccination has been implemented in Japan among tuberculin-negative first grade primary and first grade junior high school students for decades. Controversies regarding the effectiveness of BCG revaccination and low incidence of tuberculosis (TB) among Japanese children prompted this study. METHODS: Cost-effectiveness and cost-benefit analyses were conducted for a cohort of schoolchildren who underwent revaccination during 1996. The study population was a hypothetical cohort comprising 1.35 million first grade primary school and 1.51 million first grade junior high school students enrolled in 1996 at locations throughout Japan. Assuming 50% vaccine efficacy for revaccination, a 10-year duration of protection, and 5% annual discount rate, we calculated the total hypothetical number of TB cases averted, the cost and number of immunizations per TB case averted, and the benefit-cost ratio for the program. RESULTS: The revaccination program for 1996 schoolchildren cohort would prevent 296 TB cases over a 10-year period at a cost of US$ 108,378 per case averted. About 4,963 immunizations would be required to prevent one child from developing TB. The benefit-cost ratio remained at 0.13 with baseline assumptions and ranged from 0.05 to 0.29 and from 0.02 to 0.74 for one-way and two-way sensitivity analyses, respectively. CONCLUSION: BCG revaccination among schoolchildren is not supported by available scientific and economic data. Based on the results of this study, current BCG revaccination policies in Japan and other countries should be reexamined. PMID- 12079444 TI - Evaluation of a cancer prevention and detection curriculum for medical students. AB - BACKGROUND: Undergraduate medical education needs revision to ensure that medical students graduate with the skills necessary to assist their patients in cancer prevention and detection. We sought to implement and incorporate a cancer education curriculum into the students' core curriculum and to assess their skill levels prior to (1996), during (1997), and at the peak (1998-1999) of the incorporation of new hours. METHODS: We conducted pretest and posttest surveys of students at Boston University School of Medicine (medical student years 1-4) enrolled in each of the four study years (1996-1999). A total of 1,956 surveys (response rate, 82%) were completed. The primary outcome measure was the student's self-reported skill level (with responses ranging from 1 (very unskilled) to 5 (very skilled)) for counseling for tobacco cessation, tobacco prevention, and sun protection and for the early detection of breast, skin, and cervical cancer. Mean scores were computed for each chronological year of the study and medical school year. Differences and trends over time in mean scores of students in each medical school year were evaluated using multiple regression analysis. RESULTS: The number of hours of cancer education increased from 6 in 1996 to 15 in 1999. Strong improvements in self-rated skill levels were recorded for four of the six measures. In particular, tobacco cessation counseling skill rose from 2.16 (1996) to 3.13 (1999) for second year students (P < 0.001) and from 3.27 (1996) to 4.17 (1999) for fourth year students (P < 0.001). Among fourth year students, the percentage reporting that cancer prevention was given too little emphasis declined from 62% (1996) to 26% (1999) (P < 0.001), suggesting that the expanded curriculum reflected the students' preferences. CONCLUSIONS: Cancer education can be interwoven into the existing medical school curriculum and produce improvements in students' skill levels for counseling and examinations. Strategies to enhance prevention teaching can use this model. PMID- 12079445 TI - Costs and cost-effectiveness of a clinical intervention to increase mammography utilization in an inner city public health hospital. AB - BACKGROUND: Studies have demonstrated the cost-effectiveness of screening women for breast cancer; however, the cost-effectiveness of strategies to motivate women to receive breast cancer screening has been less well studied. METHODS: A total of 196 women, aged 50 to 74, who were enrolled in a public health hospital clinic, were noncompliant with mammography screening, and had at least one routine clinic appointment during the study period (15 months) were entered into a randomized, controlled trial of a motivational intervention to increase mammography rates. Costs were captured via a modified Delphi technique, accounting records, sampling of staff time logs, and an estimation of miscellaneous and overhead costs. Summary costs were calculated using Excel spread sheets. RESULTS: Overall, 49% of women who received the intervention had a mammogram within 8 weeks of an index visit compared with 22% of control women. Calculation of the cost-effectiveness of the project showed an additional cost of $151 (1996 U.S.$) for each woman receiving the intervention and $559 for each additional woman motivated to receive a mammogram. CONCLUSIONS: Cost tracking and cost-effectiveness analysis can be done when intervening in a clinical setting, thereby allowing clinics to make informed decisions about implementing programs to increase motivation of their patients to receive screening. PMID- 12079446 TI - A novel coordination polymer containing both interdigitated 1D chains and interpenetrated 2D grids. AB - A hydro(solvo)thermal reaction between zinc perchlorate and ethyl ester of a new pyridinecarboxylate bridging ligand of approximately 17.6 A in length yields a unique coordination polymer which contains both interdigitated infinite 1D chains and interpenetrated 2D rhombohedral grids [Zn(2.5)(L)(4)(mu(3)-OH)] x (H(2)O)(5), 1, where L is 3-[[4-(4-pyridylethenyl)phenyl]ethenyl]benzoate. The 1D chains contain mu(3)-bridged hydroxy groups and have a [Zn(4)(mu(3)-OH)(2)(L)(6)] stoichiometry, while the 2D grids have a Zn(L)(2) formula and diagonal distances of 31.7 and 25.2 A. Crystal data for 1: monoclinic space group P2/c, a = 15.686(2) A, b = 12.6103(16) A, c = 38.999(5) A, beta = 98.397(2) degrees, and Z = 4. PMID- 12079448 TI - Metal oxidation promoted C-H activation in manganese complexes of N-confused porphyrin. AB - The N-confused porphyrin complex Mn(II)(NCHPP)Br exhibits a nonplanar porphyrin ring with an inner core C-H on the inverted pyrrole ring. The aerobic metal oxidation promotes the dissociation of an inner-core proton on the inverted pyrrole ring and changes the N-confused porphyrin conformation to a planar porphyrin ring to form Mn(III)(NCPP)Br. The conjugate systems and metal oxidation states are confirmed by crystal structures as well as spectroscopic data. The reverse reaction can be achieved by treating the Mn(III)(NCPP)Br with p toluenesulfonhydrazide. PMID- 12079447 TI - Synthesis and DNA-binding properties of cationic 2,2':6',2' ' terpyridineplatinum(II) complexes containing 1,2- and 1,7-dicarba-closo dodecaborane(12). AB - The first examples of DNA metallointercalators containing a dicarba-closo dodecaborane(12) (carborane) moiety are presented. Treatment of the labile platinum(II) complex [Pt(OTf)(terpy)](+) (terpy = 2,2':6',2' '-terpyridine) with the 1,2-carborane monothiol derivatives 1-HS(CH(2))(n)-1,2-C(2)B(10)H(11) (n = 0, 1) or the novel 1,7-carborane ligand, 1-HSCH(2)-1,7-C(2)B(10)H(11), affords the stable, brightly colored species [Pt(1-S(CH(2))(n)-1,Z-C(2)B(10)H(11))(terpy)](+) (Z = 2, n = 0, 1; Z = 7, n = 1) in good yield and purity. Preliminary DNA-binding experiments with calf-thymus DNA indicate an intercalative interaction by the platinum(II) complexes at high r(f) values. PMID- 12079449 TI - Synthesis and characterization of technetiumtetrakis(acetonitrile)bis(triphenylphosphine) cationic complexes. AB - A new route to low-valent technetium complexes containing multiple acetonitrile ligands has been developed. The reduction of TcCl(4)(PPh(3))(2) with zinc metal dust in acetonitrile results in the formation of [Tc(CH(3)CN)(4)(PPh(3))(2)][Zn(2)Cl(6)](1/2). The hexafluorophosphate salt of the analogous Tc(II) cation can be prepared via chemical oxidation of the Tc(I) species, and the Tc(I) cation can be regenerated via chemical reduction. The compounds have been characterized in the solid state via single-crystal X-ray crystallography, and in solution via a combination of spectroscopic techniques and cyclic voltammetry. The structural parameters found in the two complexes are similar to each other; however, the difference in oxidation state is reflected, as expected, in the spectroscopic results. The electrochemical data, obtained from cyclic voltammograms of Tc(CH(3)CN)(4)(PPh(3))(2)](PF(6))(n)() (n = 1,2), mirror the synthetic results in that both compounds possess a reversible redox couple at -0.55 V versus ferrocene, which has been assigned to the Tc(II)/Tc(I) couple. PMID- 12079450 TI - Flexibility in the coordination chemistry of the 2,3-dimethylindolide ligand with potassium, yttrium, and samarium. AB - The coordination chemistry of the 2,3-dimethylindolide anion (DMI), (Me(2)C(8)H(4)N)(-), with potassium, yttrium, and samarium ions is described. In the potassium salt [K(DMI)(THF)](n), 1, prepared from Me(2)C(8)H(4)NH and KH in THF, the dimethylindole anion binds and bridges potassium ions in three different binding modes, namely eta(1), eta(3), and eta(5), to form a two-dimensional extended structure. In the dimethoxyethane (DME) adduct [K(DMI)(DME)(2)](2), 2, prepared by crystallizing a sample of 1 from DME, DMI exists as a mu eta(1):eta(1) ligand. Compound 1 reacts with SmI(2)(THF)(4) in THF to form the distorted octahedral complex trans-(DMI)(2)Sm(THF)(4), 3, in which the dimethyindolide anions are bound in the eta(1) mode to samarium. Reaction of 2,3 dimethylindole with Y(CH(2)SiMe(3))(3)(THF)(2) afforded the amide complex (DMI)(3)Y(THF)(2), 4, in which the dimethylindolide anions are also bound in the eta(1) mode to yttrium. Compound 1 also reacts with (C(5)Me(5))(2)LnCl(2)K(THF)(2) (Ln = Sm, Y) to form unsolvated amide complexes (C(5)Me(5))(2)Ln(DMI) (Ln = Sm, 5; Y, 6), in which DMI attaches primarily through nitrogen, although the edge of the arene ring is oriented toward the metals at long distances. PMID- 12079452 TI - Synthesis, crystal structure, and photoelectric properties of Re(CO)(3)ClL (L = 2 (1-ethylbenzimidazol-2-yl)pyridine). AB - A novel Re(I) complex, Re(CO)(3)ClL (L = 2-(1-ethylbenzimidazol-2-yl)pyridine), has been synthesized and structurally characterized by single-crystal X-ray diffraction analysis. Crystal data for C(17)H(13)ClN(3)O(3)Re: space group, orthorhombic, Pbca; a = 12.713(6) A; b = 15.103(7) A; c = 18.253(8) A; Z = 8. Stable vacuum vapor deposition of the Re complex has been verified by UV-vis and infrared spectroscopy. A two-layer electroluminescent device with configuration of ITO/TPD/Re(CO)(3)ClL/Mg(0.9)Ag(0.1)/Ag has been fabricated, which gave a turn on voltage of as low as 3 V and a maximum luminance of 113 cd/m(2) at a bias voltage of 10.5 V, and confirmed that the Re complex can function as a bright orange-red emitter and an electron transport material in an electroluminescent device. PMID- 12079451 TI - Redox potential modulation in mixed sandwich pyrrolyl/ dicarbollide complexes. AB - Carbon cluster (C(c)) substituents have been shown to be of essential importance in C(c).C(c) distance, rotational energy barriers, and (11)B[(1)H] NMR chemical shift values in mixed pyrrolyl/dicarbollide cobalt complexes. In the present work, the influence of electronic properties of exo-cluster substituents upon redox potential values associated to the metallic central atom in mixed pyrrolyl/dicarbollide and dimethylpyrrolyl/dicarbollide cobalt complexes is discussed. With that purpose, two new neutral sandwich species, closo-[3 Co(eta(5)-NC(4)(CH(3))(2)H(2))-1,2-(C(6)H(5))(2)-1,2-C(2)B(9)H(9)] (2) and closo [3-Co(eta(5)-NC(4)(CH(3))(2)H(2))-1-CH(3)-2-SCH(3)-1,2-C(2)B(9)H(9)] (3), have been synthesized and characterized by (1)H, (11)B, (11)B[(1)H], and (13)C[(1)H] NMR and IR spectroscopies, elemental analysis, and X-ray diffraction analysis. The redox potential (E(1/2)) of these complexes has been measured in nonpolar media and compared to values obtained for previously reported mixed complexes, incorporating alkyl, phenyl, thiophenyl, and thiomethyl exo-cluster substituents. The potential shift arising from the effect of these substituents has been discussed in terms of individual and average contribution. This last point is in the case of two identical substituting groups placed on both C(c) atoms, in which the contribution of the second introduced substituent has shown to be lower than that for the first one. The potential shift arising from the presence of methyl units on the pyrrolyl anion has also been determined. PMID- 12079453 TI - Synthesis of heterometallic bismuth/molydenum alkoxides and their behavior on silica surfaces. AB - The reaction of [(C(3)H(5))Mo(CO)(2)(CH(3)CN)(2)Cl], 2, with [Bi(OCH(2)CH(2)OCH(3))(3)](2) on a large scale leads to the novel molybdenum/bismuth alkoxide [(C(3)H(5))Mo(CO)(2)(mu-kappa O,2 kappa O' OCH(2)CH(2)OCH(3))(2)(mu-kappa O-OCH(2)CH(2)OCH(3))BiCl], 6, as the main product as well as to [(C(3)H(5))Mo(CO)(2)(mu-kappa O,2 kappa O'-OCH(2)CH(2)OCH(3))(2)(mu Cl)BiCl], 4, as a byproduct. Both compounds were characterized by elemental analysis, IR, and NMR spectroscopy as well as by X-ray diffraction. If 6 is brought into contact with a large excess of silica gel, aggregation and condensation reactions are initiated, which led to clusters of ca. 200 nm size spread over the silica surface. When the resulting material is calcinated at 350 degrees C in the presence of O(2), all organic ligands are eliminated and the metal oxo units rearrange: SEM/EDX measurements showed afterward Mo-free bismuth oxo clusters with sizes between 30 and 1000 nm, which are distributed together with molybdenum oxo particles of lower nuclearity over the silica surface. If such a material is employed as a potential catalyst for the propene oxidation under technical conditions, no activity is observed. If, however, the process is performed under very low pressures, a conversion of 5% is found. This result is discussed in the context of the mechanism proposed for the technical oxidation of propene to acrolein on bismuthmolybdate catalysts. PMID- 12079454 TI - Synthesis and structural and spectroscopic characterization of a complex between Co(II) and imino-bis(methylphosphonic acid): gaining insight into biologically relevant metal-ion phosphonate interactions or looking at a new Co(II) organophosphonate material? AB - Cobalt is an essential metal ion involved in key biomolecules, regulating processes in human physiology. As a metal ion, Co(II) assumes forms, which are dictated by the nature of organic binders in biological fluids, and the conditions under which metal ion ligand interactions arise. Among the various low molecular mass metal ion binders in biological fluids are amino acids, organic acids, as well as their variably phosphorylated forms. As a representative metal ion binder, the organophosphonate ligand H(2)O(3)P-CH(2)-NH(2)(+)-CH(2)-PO(3)H(-) was employed in aqueous reactions with Co(II), ultimately leading to the isolation of complex [Co(C(2)H(8)O(6)NP(2))(2)(H(2)O)(2)] (1) at pH 2. The complex was characterized analytically, spectroscopically (FTIR, UV-vis, EPR), and magnetically. Compound 1 crystallizes in the monoclinic space group P2(1)/n, with a = 7.361(3) A, b = 8.133(3) A, c = 14.078(5) A, beta = 104.40(1) degrees, V = 816.3(5) A(3), and Z = 2. X-ray crystallography reveals that 1 is a compound with a molecular type of lattice. In it, there exist mononuclear octahedral sites of Co(II) surrounded by oxygens, belonging to terminal phosphonates and bound water molecules. Both ends of the ligand zwitterionic form are involved in binding to adjacent Co(II) ions, thus creating tetranuclear 32-membered rings, with cavities extending in two dimensions and holes in the third dimension throughout the lattice of 1. Similar structural features were observed in other metal organophosphonate lattices of potential catalytic and chemical reactivity. The magnetic and EPR data on 1 support the presence of a high-spin octahedral Co(II) in an oxygen environment, having a ground state with an effective spin S = (1)/(2). The solution UV-vis and EPR data suggest retention of the high-spin octahedral Co(II) ion, consistent with the magnetization measurements on 1. Collectively, the data reflect the existence of a soluble Co(II) iminodiphosphonate species not unlike those expected in biological fluids containing the specific ligand or ligands similar to that. Both biologically relevant perspectives and a synthetic outlook into Co(II)-organophosphonate materials are discussed. PMID- 12079455 TI - Synthesis, structures, and magnetic properties of a series of lanthanum(III) and gadolinium(III) complexes with chelating benzimidazole-substituted nitronyl nitroxide free radicals. Evidence for antiferromagnetic Gd(III)-radical interactions. AB - This paper reports the synthesis, crystal structures, and magnetic properties of a series of lanthanide complexes with nitronyl nitroxide radicals of general formula [[Ln(III)(radical)(4)] x (ClO(4))(3) x (H(2)O)(x) x (THF)(y)] (1-4) and [Ln(III)(radical)(2)(NO(3))(3)] (5, 6) [Ln = La (compounds 1, 3, 5) or Gd (compounds 2, 4, and 6); radical = 2-(2'-benzymidazolyl)-4,4,5,5 tetramethylimidazoline-1-oxyl-3-oxide (NITBzImH, compounds 1, 2, 5, 6) or 2-[2' [(6'-methyl)benzymidazolyl]]-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (NITMeBzImH, compounds 3, 4)]. (1) C(64)H(88)Cl(3)LaN(16)O(24), fw = 1710.76, orthorhombic, Fddd, a = 11.0682(8) A, b = 34.240(3) A, c = 42.787(3) A, V = 16215(2) A(3), Z = 8, R = 0.0876, R(w) = 0.2336. (2) C(64)H(88)Cl(3)GdN(16)O(24), fw = 1729.10, tetragonal, P 4 macro 2c, a = 16.0682(4) A, b = 16.0682(4) A, c = 18.7190(6) A, V = 4833.0(2) A(3), R = 0.0732, R(w) = 0.2218. (3) C(68)H(94)Cl(3)LaN(16)O(23), fw = 1742.80, tetragonal, P 4 macro 2(1)m, a = 21.125(3) A, b = 21.125(3) A, c = 10.938(2) A, V = 4881.5(14) A(3), R = 0.1017, R(w) = 0.3126. (5) C(28)H(34)LaN(11)O(13), fw = 871.57, orthorhombic, Pna2(1), a = 19.5002(12) A, b = 13.0582(8) A, c = 14.5741(9) A, V = 3711.1(4) A(3), R = 0.0331, R(w) = 0.1146. (6) C(28)H(34)GdN(11)O(13), fw = 889.91, orthorhombic, Pna2(1), a = 19.1831(10) A, b = 13.1600(7) A, c = 14.4107(7) A, V = 3638.0(3) A(3), Z = 4, R = 0.0206, R(w) = 0.0625. Compounds 1-4 consist of [M(III)(radical)(4)](3+) cations, uncoordinated perchlorate anions, THF, and water crystallization molecules. In these complexes, the coordination number around the lanthanide ion is eight, and the polyhedron is either a distorted dodecahedron (1) or a distorted cube (2, 3). The crystal structures of 5 and 6 consist of independent [M(III)(radical)(2)(NO(3))(3)] entities in which the lanthanide is ten-coordinated and has a distorted bicapped square antiprism coordination polyhedron. For the lanthanum(III) complexes, the temperature dependence of the magnetic susceptibility indicates that radical-radical magnetic interactions are negligible either for compounds 1 and 3, while for compound 5 it is simulated considering dimers of weakly antiferromagnetically coupled radicals (J(rad-rad) = -1.1 cm(-1)). In the case of the gadolinium(III) compounds (2, 4, 6), each magnetic behavior gives unambiguous evidence of antiferromagnetic Gd(III)-radical interaction (2, J(Gd-rad) = -1.8 cm(-1); 4, J(Gd-rad) = -3.8 cm( 1); 6, J(Gd-rad1) = -4.05 cm(-1) and J(Gd-rad2) = -0.80 cm(-1)), in contrast to the ferromagnetic case generally observed. The nature of the Gd(III)-radical interaction is explained in relation to the donor strength of the free radical ligand. PMID- 12079456 TI - Formation of metal-anion arrays within layered perovskite hosts. Preparation of a series of new metastable transition-metal oxyhalides, (MCl)LaNb(2)O(7) (M = Cr, Mn, Fe, Co). AB - A new series of transition-metal oxyhalides (MCl)LaNb(2)O(7) (M = Cr, Mn, Fe, Co) have been prepared by a simple topochemical route. Layered perovskite hosts (ALaNb(2)O(7), A = Li, Na, K or Rb) were reacted with the corresponding anhydrous metal halides under mild reaction conditions (<400 degrees C). The compounds were examined by X-ray powder diffraction; the series appears to be isostructural with (CuCl)LaNb(2)O(7), and the layer spacings, with the exception of M = Co, follow the trend expected from transition-metal cationic radii. Thermal analysis with differential scanning calorimetry (DSC) shows the materials to be metastable where all four compounds decompose exothermically above 690 degrees C. PMID- 12079457 TI - A novel copper cobalt inorganometallic cluster. Synthesis, structure and bonding analysis of Cu(3)[mu(2)-(CCHCo(2)(CO)(6))](3). AB - The reaction of the organometallic carboxylic acid HOOCCCHCo(2)(CO)(6) with copper(II) methoxide leads to a new inorganometallic cluster; Cu(3)[mu(2) (CCHCo(2)(CO)(6))](3). This cluster has a triangular core of copper(I) centers surrounded by three CCHCo(2)(CO)(6) fragments. The structure of the cluster has short Cu-Cu and Cu-Co distances (average 2.500 and 2.540 A, respectively). DFT calculations provide a rationalization of the peculiar bonding in this cluster. PMID- 12079458 TI - Synthesis of heteroleptic anthryl-substituted beta-ketoenolates of rhodium(III) and iridium(III): photophysical, electrochemical, and EPR study of the fluorophore-metal interaction. AB - The anthryl-substituted rhodium(III) and iridium(III) heteroleptic beta ketoenolato derivatives of general formula [M(acac)(2)(anCOacac)] [acac = pentane 2,4-dionate; anCOacac = 3-(9-anthroyl)pentane-2,4-dionate], 3 (M = Rh) and 4 (M = Ir), and [M(acac)(2)(anCH(2)acac)] [anCH(2)acac = 3-(9-anthrylmethyl)pentane-2,4 dionate], 5 (M = Rh) and 6 (M = Ir), were prepared by reacting the corresponding tris(pentane-2,4-dionate)metal complexes, [M(acac)(3)], with 9-anthroyl chloride and 9-chloromethylanthracene, respectively, under Friedel-Crafts conditions. 3-6 were characterized by elemental analysis, ion spray mass spectrometry (IS-MS), (1)H NMR, and UV-vis spectroscopy. The structure of 3 was also elucidated by single-crystal X-ray analysis. When excited at 365 nm, 3-6 result to be poorly luminescent compounds; while the free diketone, i.e., 3-(9-anthrylmethyl)pentane 2,4-dione 1, whose structure was established also by single-crystal X-ray analysis, results to be a strongly light emitting molecule. The study of the electrochemical behavior of 3-6 as well as of the corresponding tris acetylacetonates of rhodium(III) and iridium(III) allows a satisfactory interpretation of their electrode process mechanism, and gives information about the location of the redox sites along with the thermodynamic and kinetic characterization of the corresponding redox processes. All data are in agreement with the hypothesis that the quenching of the anthracene fluorescence, observed for compounds 3-6, can be due to an intramolecular electron transfer process between the anthryl moiety and the metal-beta-ketoenolato component. Moreover, a study was carried out of the redox behavior of the dyads 3-6 under chemical activation. The one-electron oxidation of compounds 3-6 by thallium(III) trifluoroacetate leads to the formation of the corresponding cation radicals, 3(+)-6(+), whose highly resolved X-band EPR spectra were fully interpreted by computer simulation as well as by semiempirical and DFT calculations of spin density distribution. PMID- 12079459 TI - Construction of hydrogen-bonded and coordination-bonded networks of cobalt(II) with pyromellitate: synthesis, structures, and magnetic properties. AB - Synthesis (hydrothermal and metathesis), characterization (UV-vis, IR, TG/DTA), single-crystal X-ray structures, and magnetic properties of three cobalt(II) pyromellitate complexes, purple [Co(2)(pm)](n) (1), red [Co(2)(pm)(H(2)O)(4)](n) x 2nH(2)O (2), and pink [Co(H(2)O)(6)](H(2)pm) (3) (H(4)pm = pyromellitic acid (1,2,4,5-benzenetetracarboxylic acid)), are described. 1 consists of one dimensional chains of edge-sharing CoO(6) octahedra that are connected into layers via O-C-O bridges. The layers are held together by the pyromellitate (pm(4 )) backbone to give a three-dimensional structure, each ligand participating in an unprecedented 12 coordination bonds (Co-O) to 10 cobalt atoms. 2 consists of a three-dimensional coordination network possessing cavities in which unbound water molecules reside. This highly symmetric network comprises eight coordinate bonds (Co-O) between oxygen atoms of pm(4-) to six trans-Co(H(2)O)(2). 3 possesses a hydrogen-bonded sandwich structure associating layers of [Co(H(2)O)(6)](2+) and planar H(2)pm(2-). The IR spectra, reflecting the different coordination modes and charges of the pyromellitate, are presented and discussed. The magnetic properties of 1 indicate complex behavior with three ground states (collinear and canted antiferromagnetism and field-induced ferromagnetism). Above the Neel temperature (T(N)) of 16 K it displays paramagnetism with short-range ferromagnetic interactions (Theta = +16.4 K, mu(eff) = 4.90 mu(B) per Co). Below T(N) a weak spontaneous magnetization is observed at 12.8 K in low applied fields (H < 100 Oe). At higher fields (H > 1000 Oe) metamagnetic behavior is observed. Two types of hysteresis loops are observed; one centered about zero field and the second about the metamagnetic critical field. The critical field and the hysteresis width increase as the temperature is lowered. The heat capacity data suggest that 1 has a 2D or 3D magnetic lattice, and the derived magnetic entropy data confirm an anisotropic s(eff) = 1/2 for the cobalt(II) ion. Magnetic susceptibility data indicate that 2 and 3 are paramagnets. PMID- 12079460 TI - Bi-1,10-phenanthrolines and their mononuclear Ru(II) complexes. AB - A series of four biphen (phen = 1,10-phenanthroline) ligands, 2,2'-biphen (1), 3,3'-biphen (2), 2,2'-dimethylene-3,3'-biphen (3), and 2,3'-dimethylene-3,2' biphen (4), is prepared by coupling and Friedlander methodology. The corresponding mononuclear Ru(II) complexes, [Ru(1-4)(Mebpy)(2)](2+) where Mebpy = 4,4'-dimethyl-2,2'-bipyridine, are prepared. These complexes show long wavelength electronic absorptions at 441-452 nm and emissions at 622-641 nm. Metal-based oxidations occur in the range 1.18-1.21 V, and ligand-based reductions, at -1.20 to -1.30 V. The addition of Zn(2+), Cd(2+), or Hg(2+) ions results in a strong enhancement and red shift of the luminescence of complex Ru-3. Alkali and alkaline earth metal ions barely affect the luminescence of Ru-3 while transition metal ions such as Co(2+), Cu(2+), Ni(2+), and Mn(2+) lead to efficient quenching of the Ru-3 luminescence. The luminescence of Ru-2 and Ru-4 is quenched in the presence of Zn(2+) because of a conformationally induced reduction in electronic communication between the two phen halves of the ligand. The addition of Zn(2+) has only a slight effect on the luminescence of Ru-1 because of steric hindrance toward complexation. PMID- 12079461 TI - A study of structural and bonding variations in the homologous series [Mo(2)(CN)(6)(dppm)(2)](n-) (n = 2, 1, 0). AB - Reaction of Mo(2)Cl(4)(dppm)(2) (dppm = bis(diphenylphosphino)methane) with 6 equiv of [n-Bu(4)N][CN] or [Et(4)N][CN] in dichloromethane yields [n Bu(4)N](2)[Mo(2)(CN)(6)(dppm)(2)] (1) and [Et(4)N](2)[Mo(2)(CN)(6)(dppm)(2)] (2), respectively. The corresponding one- and two-electron oxidation products [n Bu(4)N][Mo(2)(CN)(6)(dppm)(2)] (3) and Mo(2)(CN)(6)(dppm)(2) (4)were prepared by reactions of 1 with the oxidant NOBF(4). Single-crystal X-ray structures of 2.2CH(3)CN, 3.2CH(3)CN.2H(2)O, and 4.2CH(3)NO(2) were performed, and the results confirmed that all three complexes contain identical ligand sets with trans dppm ligands bisecting the Mo(2)(mu-CN)(2)(CN)(4) equatorial plane. The binding of the bridging cyanide ligands is affected by the oxidation state of the dimolybdenum core as evidenced by an increase in side-on pi-bonding overlap of the mu-CN in going from 1 to 4. The greater extent of pi-donation into Mo orbitals is accompanied by a lengthening of the Mo-Mo distance (2.736(1) A in Mo(2)(II,II) (2), 2.830(1) A in Mo(2)(II,III) (3), and 2.936(1) A in Mo(2)(III,III) (4)). A computational study of the closed-shell members of this homologous series, [Mo(2)(CN)(6)(dppm)(2)](n)() (n = 2-, 0), indicates that the more pronounced side on pi-donation evident in the X-ray structure of 4 leads to significant destabilization of the delta orbital and marginal stabilization of the delta() orbitals with respect to nearly degenerate delta and delta orbitals in the parent compound, 2. The loss of delta contributions combined with the reduced orbital overlap due to higher charges on molybdenum centers in oxidized complexes 3 and 4 is responsible for the observed increase in the length of the Mo-Mo bond. PMID- 12079462 TI - Tristhiolatomolybdenum nitrides, (RS)(3)Mo[triple bond]N where R = (i)Pr and (t)Bu, preparation, characterization and comparisons with related trialkoxymolybdenumnitrides. AB - The addition of thiols to ((t)BuO)(3)Mo[triple bond]N in toluene leads to the formation of (RS)(3)Mo[triple bond]N compounds as yellow, air-sensitive compounds, where R = (i)Pr and (t)Bu. The single-crystal structure of ((t)BuS)(3)Mo[triple bond]N reveals a weakly associated dimeric structure where two ((t)BuS)(3)Mo[triple bond]N units (Mo-N = 1.61 A, Mo-S = 2.31 A (av)) are linked via thiolate sulfur bridges with long 3.03 A (av) Mo-S interactions. Density functional theory calculations employing Gaussian 98 B3LYP (LANL2DZ for Mo and 6-31G* for N, O, S, and H) have been carried out for model compounds (HE)(3)Mo[triple bond]N and (HE)(3)MoNO, where E = O and S. A comparison of the structure and bonding within the related series ((t)BuE)(3)Mo[triple bond]N and ((t)BuE)(3)MoNO is made for E = O and S. In the thiolate compounds, the highest energy orbitals are sulfur lone-pair combinations. In the alkoxides, the HOMO is the N 2p lone-pair which has M-N sigma and M-O pi* character for the nitride. As a result of greater O p pi to Mo pi interactions, the M-N pi orbitals of the Mo-N triple bond are destabilized with respect to their thiolate counterpart. For the nitrosyl compounds, the greater O p pi to Mo d pi interaction favors greater back bonding to the nitrosyl pi* orbitals for the alkoxides relative to the thiolates. The results of the calculations are correlated with the observed structural features and spectroscopic properties of the related alkoxide and thiolate compounds. PMID- 12079463 TI - Tuning the electronic structure of octahedral iron complexes [FeL(X)] (L = 1 alkyl-4,7-bis(4-tert-butyl-2-mercaptobenzyl)-1,4,7-triazacyclononane, X = Cl, CH(3)O, CN, NO). The S = 1/2 <==>3/2 Spin equilibrium of [FeL(Pr)(NO)]. AB - Two new pentadentate, pendent arm macrocyclic ligands of the type 1-alkyl-4,7 bis(4-tert-butyl-2-mercaptobenzyl)-1,4,7-triazacyclononane where alkyl represents an isopropyl, (L(Pr))(2-), or an ethyl group, (L(Et))(2-), have been synthesized. It is shown that they bind strongly to ferric ions generating six-coordinate species of the type [Fe(L(alk))X]. The ground state of these complexes is governed by the nature of the sixth ligand, X: [Fe(III)(L(Et))Cl] (2) possesses an S = 5/2 ground state as do [Fe(III)(L(Et))(OCH(3))] (3) and [Fe(III)(L(Pr))(OCH(3))] (4). In contrast, the cyano complexes [Fe(III)(L(Et))(CN)] (5) and [Fe(III)(L(Pr))(CN)] (6) are low spin ferric species (S = 1/2). The octahedral [FeNO](7) nitrosyl complex [Fe(L(Pr))(NO)] (7) displays spin equilibrium behavior S = 1/2<==>S = (3)/(2) in the solid state. Complexes [Zn(L(Pr))] (1), 4.CH(3)OH, 5.0.5toluene.CH(2)Cl(2), and 7.2.5CH(2)Cl(2) have been structurally characterized by low-temperature (100 K) X-ray crystallography. All iron complexes have been carefully studied by zero- and applied-field Mossbauer spectroscopy. In addition, Sellmann's complexes [Fe(pyS(4))(NO)](0/1+) and [Fe(pyS(4))X] (X = PR(3), CO, SR(2)) have been studied by EPR and Mossbauer spectroscopies and DFT calculations (pyS(4) = 2,6-bis(2 mercaptophenylthiomethyl)pyridine(2-)). It is concluded that the electronic structure of 7 with an S = 1/2 ground state is low spin ferrous (S(Fe) = 0) with a coordinated neutral NO radical (Fe(II)-NO) whereas the S = 3/2 state corresponds to a high spin ferric (S(Fe) = 5/2) antiferromagnetically coupled to an NO(-) anion (S = 1). The S = 1/2<==>S = 3/2 equilibrium is then that of valence tautomers rather than that of a simple high spin<==>low spin crossover. PMID- 12079464 TI - Polyatomic clusters of the triel elements. Palladium-centered clusters of thallium in A(8)Tl(11)Pd, A = Cs, Rb, K. AB - Reactions of the elements within welded Ta containers at approximately 600 degrees C followed by slow cooling give new A(8)Tl(11)Pd(x) products from an apparently continuous encapsulation of Pd atoms into the pentacapped trigonal prismatic anions in the isotypic rhombohedral (R3 macro c) A(8)Tl(11) phases. All systems also produce other phases at x < 1 as well, the simplest being the cesium system in which only trigonal Pd(13)Tl(9) is also formed. Cs(8)Tl(11)Pd(0.84(1)) was characterized by single-crystal means as close to the upper x limit in that system (R3 macro c, Z = 6, a = 10.610(1) A, c = 54.683(8) A). The Pd insertion causes an expansion of the D(3) host anion, particularly about the waist, to generate a trigonal bipyramidal PdTl(5) unit (d(Pd-Tl) approximately 2.6-2.8 A) centered within a somewhat larger Tl(6) trigonal prism, the remainder of the Tl(11) cluster. Strong Tl cage bonding is retained. Extended Huckel calculations show significant involvement of all Tl 6s, 6p and Pd 4d, 5s, 5p orbital sets in the central and cage bonding. The last valence electron is considered to be delocalized in a conduction band, as in A(8)Tr(11) examples, rather than occupying an antibonding e'' LUMO across a gap of approximately 2.4 eV. PMID- 12079465 TI - Carbon dioxide as a solubility "switch" for the reversible dissolution of highly fluorinated complexes and reagents in organic solvents: application to crystallization. AB - Highly fluorinated organic or organometallic solid compounds can be made to dissolve in liquid hydrocarbons by the application of 20-70 bar of CO(2) gas. Subsequently releasing the gas causes the compounds to precipitate or crystallize, giving quantitative recovery of the solid. The resulting crystals can be of sufficient quality for single-crystal X-ray crystallography; the structures of Rh(2)(O(2)CCF(2)CF(2)CF(3))(4)(DMF)(2), Rh(2)(O(2)C(CF(2))(9)F)(4)(MeOH)(2), Cr(hfacac)(3), and P[C(6)H(3)(3,5 CF(3))(2)](3) have been determined from crystals grown in this manner. PMID- 12079466 TI - Effect of steric encumbrance of tris(3-phenylpyrazolyl)borate on the structure and properties of ternary copper(II) complexes having N,N-donor heterocyclic bases. AB - Complexes of formulation [Cu(Tp(Ph))(L)](ClO(4)) (1-4), where Tp(Ph) is anionic tris(3-phenylpyrazolyl)borate and L is N,N-donor heterocyclic base, viz. 2,2' bipyridine (bpy, 1), 1,10-phenanthroline (phen, 2), dipyridoquinoxaline (dpq, 3), and dipyridophenazine (dppz, 4), are prepared from a reaction of copper(II) acetate.hydrate with KTp(Ph) and L in CH(2)Cl(2) and isolated as perchlorate salts. The complexes are characterized by analytical, structural, and spectral methods. The crystal structures of complexes 1-4 show the presence of discrete cationic complexes having the metal, Tp(Ph), and L in a 1:1:1 ratio and a noncoordinating perchlorate anion. The complexes have a square-pyramidal 4 + 1 coordination geometry in which two nitrogens of L and two nitrogens of the Tp(Ph) ligand occupy the basal plane and one nitrogen of Tp(Ph) binds at the axial site. Complexes 3 and 4 display distortion from the square-pyramidal geometry. The Cu-N distances for the equatorial and axial positions are approximately 2.0 and 2.2 A, respectively. The phenyl groups of Tp(Ph) form a bowl-shaped structure that encloses the [CuL] moiety. The steric encumbrance is greater for the bpy and phen ligands compared to that for dpq and dppz. The one-electron paramagnetic complexes (mu approximately equal to 1.8 mu(B)) exhibit axial EPR spectra in CH(2)Cl(2) glass at 77 K giving g(parallel) and g(perpendicular) values of approximately 2.18 (A(parallel) = 128 G) and approximately 2.07. The data suggest a [d(x(2)-y(2))](1) ground state. The complexes are redox-active and display a quasireversible cyclic voltammetric response for the Cu(II)/Cu(I) couple near 0.0 V versus SCE with an i(pc)/i(pa) ratio of unity in CH(2)Cl(2) or DMF-0.1 M TBAP. The E(1/2) values of the couple vary in the order 4 > 3 > 2 > 1. A profound effect of steric encumbrance caused by the Tp(Ph) ligand is observed in the reactivity of 1-4 with the calf thymus (CT) and supercoiled (SC) DNA. Complexes 2 4 show similar binding to CT DNA. The propensity for the SC DNA cleavage varies as 4 > 3 > 2. The bpy complex does not show any significant binding or cleavage of DNA. Mechanistic investigations using distamycin reveal minor groove binding for 2 and 3 and a major groove binding for 4. The scission reactions that are found to be inhibited by hydroxyl radical scavenger DMSO are likely to proceed through sugar hydrogen abstraction pathways. PMID- 12079467 TI - Controlling the framework formation of silver(I) coordination polymers with 1,4 bis(phenylthio)butane by varying the solvents, metal-to-ligand ratio, and counteranions. AB - The reactions of 1,4-bis(phenylthio)butane (L) with Ag(I) salts in varied conditions (varying the solvents, metal-to-ligand ratios, and counteranions) lead to the formation of four new two-dimensional (2D) coordination polymers with different network structures: [Ag(2)L(3)(ClO(4))(2)](infinity) 1, [Ag(2)L(3)(ClO(4))(2) x CH(3)OH](infinity) 2, [[AgL(2)](ClO(4))](infinity) 3, and [AgLNO(3)](infinity) 4. All the structures were established by single-crystal X ray diffraction analysis. Crystal data for 1: triclinic, P-1, a = 11.0253(9) A, b = 11.3455(9) A, c = 11.5231(9) A, alpha = 93.931(2) degrees, beta = 92.689(2) degrees, gamma = 112.9810(10) degrees, Z = 2. 2: triclinic, P-1, a = 11.9147(13) A, b = 16.1534(17) A, c = 16.2259(17) A, alpha = 74.977(2) degrees, beta = 69.030(2) degrees, gamma = 69.986(2) degrees, Z = 2. 3: triclinic, P-1, a = 12.1617(9) A, b = 12.5054(10) A, c = 13.1547(10) A, alpha = 64.3370(10) degrees, beta =85.938 (2) degrees, gamma = 69.3010(10) degrees, Z = 2. 4: monoclinic, P2(1)/c, a = 5.4032(17) A, b = 16.974(6) A, c = 19.489(6) A, beta = 94.234(6) degrees, Z = 4. In all four complexes, each Ag(I) center has a tetracoordination geometry, and the 2D networks consist of fused large macrometallacyclic ring systems. The "hexagonal" 42-membered rings, Ag(6)L(6), observed in 1 and 2 are nearly identical, which could be considered as unique examples of self-sustaining noninterpenetrated frameworks formed with flexible ligands. The repeating rectangular 28-membered macrometallacycle, Ag(4)L(4), is the basis for the network of 3, in which the perchlorate anions occupy the voids to prevent the ring from collapsing. In 4, columns of the fused rectangular 22-membered rings, Ag(4)L(2)(NO(3))(2), are cross-linked through the L ligands to form a unique 2D network consisting of two types of 22-membered repeating units. PMID- 12079468 TI - Microspeciation in the copper(II)-L-histidylglycine system. An ESR study by the two-dimensional computer simulation method. AB - Twelve ESR-active (and one inactive) copper(II) complexes of L-histidylglycine (HL) were characterized via their formation (micro)constants and ESR parameters obtained by two-dimensional ESR spectroscopic evaluation in aqueous solution. In strongly acidic media, the ligand is coordinated through its N-terminal donor groups: the complex [CuLH(2)](3+) involves monodentate imidazole binding, whereas [CuLH](2+) involves bidentate ligation through the amino and imidazole N atoms. This histamine-like bonding mode also predominates in the isomers of [CuL(2)], formed at ligand excess near pH 7: in the major 4N isomer, both ligands occupy two equatorial sites, while in the 3N isomer, the second dipeptide is coordinated equatorially by the amino and axially by the imidazole groups. At above pH 3-4, deprotonation of the peptide group also starts: in approximately 60% of the molecules of [CuL](+), the peptide group is deprotonated, while in the minor isomer histamine-like coordination occurs. At higher pH, the active dimer [Cu(2)L(2)H(-2)], the mixed hydroxo complexes (the inactive [Cu(2)L(2)H(-3)](-) and the active [CuLH(-2)](-)), and the bis complexes [CuL(2)H](+) and [CuL(2)H( 1)](-) all involve tridentate equatorial ligation of the backbone by the amino and deprotonated peptide N and the carboxylate O atoms. In the active dimer, the neutral imidazole groups form bridges between CuLH(-1) units. In [CuL(2)H](+), the second ligand is bound equatorially via its imidazole group; in [CuL(2)H( 1)](-), the L ligand occupies the fourth equatorial site and an axial site through its amino and imidazole N atoms, respectively. PMID- 12079469 TI - Five-coordinate complexes [FeX(depe)(2)]BPh(4), X = Cl, Br: electronic structure and spin-forbidden reaction with N(2). AB - The bonding of N(2) to the five-coordinate complexes [FeX(depe)(2)](+), X = Cl (1a) and Br (1b), has been investigated with the help of X-ray crystallography, spectroscopy, and quantum-chemical calculations. Complexes 1a and 1b are found to have an XP(4) coordination that is intermediate between square-pyramidal and trigonal-bipyramidal. Mossbauer and optical absorption spectroscopy coupled with angular overlap model (AOM) calculations reveal that 1a and 1b have (3)B(1) ground states deriving from a (xz)(1)(z(2))(1) configuration. The zero-field splitting for this state is found to be 30-35 cm(-1). In contrast, the analogous dinitrogen complexes [FeX(N(2))(depe)(2)](+), X = Cl (2a) and Br (2b), characterized earlier are low-spin (S = 0; Wiesler, B. E.; Lehnert, N.; Tuczek, F.; Neuhausen, J.; Tremel, W. Angew. Chem, Int. Ed. 1998, 37, 815-817). N(2) bonding and release in these systems are thus spin-forbidden. It is shown by density functional theory (DFT) calculations of the chloro complex that the crossing from the singlet state (ground state of 2a) to the triplet state (ground state of 1a) along the Fe-N coordinate occurs at r(C) = 2.4 A. Importantly, this intersystem crossing lowers the enthalpy calculated for N(2) release by 10-18 kcal/mol. The free reaction enthalpy Delta G degrees for this process is calculated to be 4.7 kcal/mol, which explains the thermal instability of N(2) complex 2a with respect to the loss of N(2). The differences in reactivity of analogous trans hydrido systems are discussed. PMID- 12079470 TI - Electronic structure of the alpha and beta isomers of [Mo(8)O(26)](4-). AB - The structure and bonding in alpha and beta octamolybdate anions have been investigated using density functional methods. In general, good computational experimental agreement for the geometrical parameters has been obtained. The electronic structure of the anions has been probed with molecular orbital and Mulliken-Mayer methods. All Mo-O interactions have been found to be predominantly d(Mo)-p(O) in character. Several multicentered molecular orbitals can be described as sigma or pi closed-loop structures, but the proposed connection with the stability of the polyanions is not completely supported by the calculations. Mayer indexes correspond to fractional multiple character for terminal bonds and approximately single or low-order character for bridging bonds, in accordance with structural and bond valence results. The valency analysis has yielded similar overall bonding capacity for the various oxygen atoms. A distribution of the negative charge over all types of oxygen sites and metal charges considerably smaller than the formal oxidation states have been obtained from the Mulliken analysis. PMID- 12079471 TI - Antiferromagnetic coupling in a six-coordinate high spin cobalt(II)-semiquinonato complex. AB - The 3,5-di-tert-butyl-catecholato and 9,10-phenanthrenecatecholato adducts of the cobalt-tetraazamacrocycle complex Co(Me(4)cyclam)(2+) (Me(4)cyclam = 1,4,8,11 tetramethyl-1,4,8,11-tetraazacyclotetradecane) were synthesized and oxidized. The oxidation reaction products were isolated in the solid state as hexafluorophosphate derivatives. Both these complexes can be formulated as 1:1 cobalt(II)-semiquinonato complexes, that is, Co(Me(4)cyclam)(DBSQ)PF(6) (1) and Co(Me(4)cyclam)(PhSQ)PF(6) (2), in the temperature range 4-300 K, in striking contrast with the charge distribution found in similar adducts formed by related tetraazamacrocycles. The synthesis strategy and the structural, spectroscopic, and magnetic properties are reported and discussed. The crystallographic data for 2 are as follows: monoclinic, space group P2(1)/a, nomicron. 14, a = 14.087(4) A, b = 15.873(4) A, c = 14.263 (7) A, alpha = 89.91(3) degrees, beta = 107.34(2) degrees, gamma = 90.08(2) degrees, Z = 4. Both these complexes are characterized by triplet electronic ground states arising from the antiferromagnetic coupling between the high-spin d(7) metal ion and the radical ligand. PMID- 12079472 TI - Unsymmetrical tren-based ligands: synthesis and reactivity of rhenium complexes. AB - Reaction of bis(2-aminoethyl)(3-aminopropyl)amine with C(6)F(6) and K(2)CO(3) in DMSO yields unsymmetrical [(C(6)F(5))HNCH(2)CH(2)](2)NCH(2)CH(2)CH(2)NH(C(6)F(5)) ([N(3)N]H(3)). The tetraamine acts as a tridentate ligand in complexes of the type H[N(3)N]Re(O)X (X = Cl 1, Br 2) prepared by reacting Re(O)X(3)(PPh(3))(2) with [N(3)N]H(3) and an excess of NEt(3) in THF. Addition of 1 equiv of TaCH(CMe(2)Ph)Br(3)(THF)(2) to 1 gives the dimeric compound H[N(3)N]ClReOReBrCl[N(3)N]H (3) in quantitative yield that contains a Re(V)[double bond]O[bond]Re(IV) core with uncoordinated aminopropyl groups in each ligand. Addition of 2 equiv of TaCH(CMe(2)Ph)Cl(3)(THF)(2) to 1 leads to the chloro complex [N(3)N]ReCl (4) with all three amido groups coordinated to the metal, whereas by addition of 2 equiv of TaCH(CMe(2)Ph)Br(3)(THF)(2) to 2 the dibromo species H[N(3)N]ReBr(2) (5) with one uncoordinated amino group is isolated. Reduction of 4 under an atmosphere of dinitrogen with sodium amalgam gives the dinitrogen complex [N(3)N]Re(N(2)) (6). Single-crystal X-ray structure determinations have been carried out on complexes 1, 3, 5, and 6. PMID- 12079473 TI - Tetrakis(N,N'-dimethylbenzamidinato)diruthenium(III) compounds bearing axial chloro and alkynyl ligands: a new family of redox rich diruthenium compounds. AB - Novel diruthenium(III) compound Ru(2)(DMBA)(4)Cl(2) (1, DMBA = N,N' dimethylbenzamidinate) was obtained via refluxing Ru(2)(OAc)(4)Cl with dimethylbenzamidine in the presence of LiCl and Et(3)N under ambient atmosphere. Metathesis reactions between 1 and MC(2)Y (M = Li and Na) yielded bis-alkynyl derivatives Ru(2)(DMBA)(4)(C(2)Y)(2) (Y = SiMe(3) (2a), H (2b), Ph (2c), and C(2)SiMe(3) (3a)), and further desilylation of 3a using K(2)CO(3) resulted in Ru(2)(DMBA)(4)(C(4)H)(2) (3b). Compound 1 is paramagnetic (S = 1), while compounds 2 and 3 are diamagnetic. The single-crystal X-ray diffraction study revealed that the Ru-Ru distances are 2.3224(7), 2.4501(6), and 2.4559(6) A for 1, 2a, and 3b, respectively. A strong Ru-C sigma-bond in alkynyl adducts was implied by the short Ru-C distances in 2a (1.955(4) A) and 3b (1.952[5] A). All the compounds undergo three one-electron redox processes, an oxidation and two reductions, and the reversibility of redox couples depends on the nature of axial ligands. PMID- 12079474 TI - Chalcogen-rich lanthanide clusters with fluorinated thiolate ligands. AB - Mixtures of Ln(SC(6)F(5))(3) and Ln(EPh)(3) (E = S, Se) react with elemental E to give chalcogen-rich clusters with fluorinated thiolate ancillary ligands. The structures of both (THF)(6)Yb(4)S(SS)(4)(SC(6)F(5))(2) and (THF)(6)Yb(4)Se(SeSe)(4)(SC(6)F(5))(2) have been established by low-temperature single-crystal X-ray diffraction. Both compounds contain a square array of Yb(III) ions connected by a central mu(4)-E(2-) ligand. The edges of the square Yb(4) array are bridged by four mu(2)(EE) ligands, and two terminal SC(6)F(5) are on the same side of the Ln(4) plane that is capped by the mu(4)-E(2-) ion. Redox inactive (THF)(6)Tm(4)Se(SeSe)(4)(SC(6)F(5))(2) was also prepared to establish the extension of this chemistry to the redox inactive Ln. These clusters are soluble in toluene. PMID- 12079475 TI - Synthesis and characterization of mononuclear zinc aryloxide complexes supported by nitrogen/sulfur ligands possessing an internal hydrogen bond donor. AB - Treatment of a dinuclear zinc hydroxide complex ([(bmnpaZn)(2)(mu OH)(2)](ClO(4))(2) (1) or [(benpaZn)(2)(mu-OH)(2)](ClO(4))(2) (2)) with excess equivalents of an aryl alcohol derivative (p-HOC(6)H(4)X; X = NO(2), CHO, CN, COCH(3), Br, H, OCH(3)) yielded the nitrogen/sulfur-ligated zinc aryloxide complexes [(bmnpa)Zn(p-OC(6)H(4)NO(2))](ClO(4)) (3), [(benpa)Zn(p OC(6)H(4)NO(2))](ClO(4)) (4), [(benpa)Zn(p-OC(6)H(4)CHO)](ClO(4)) (5), [(benpa)Zn(p-OC(6)H(4)CN)](ClO(4)) (6), [(benpa)Zn(p-OC(6)H(4)COCH(3))](ClO(4)) x 0.5H(2)O (7), [(benpa)Zn(p-OC(6)H(4)Br)](ClO(4)) (8), [(benpa)Zn(p OC(6)H(5))](ClO(4)) (9), and [(benpa)Zn(p-OC(6)H(5)OCH(3))](ClO(4)) (10). The solid state structures of 2, 3, 5, and 6 have been determined by X-ray crystallography. While 3 and 6 exhibit a mononuclear zinc ion possessing a distorted five-coordinate trigonal bipyramidal geometry, in 5 each zinc center exhibits a distorted six-coordinate octahedral geometry resulting from coordination of the aldehyde carbonyl oxygen of another zinc-bound aryloxide ligand, yielding a chain-type structure. Zinc coordination of the aldehyde carbonyl of 5 is indicated by a large shift (>40 cm(-)(1)) to lower energy of the carbonyl stretching vibration (nu(C[double bond]O) in solid state FTIR spectra of the complex. In the solid state structures of 3, 5, and 6, a hydrogen-bonding interaction is found between N(3)-H of the supporting bmnpa/benpa ligand and the zinc-bound oxygen atom of the aryloxide ligand (N(3)...O(1) approximately 2.78 A). Solution (1)H and (13)C NMR spectra of 3-10 in CD(3)CN and FTIR spectra in CH(3)CN are consistent with all of the aryloxide complexes having a similar solution structure, with retention of the hydrogen-bonding interaction involving N(3)-H and the oxygen atom of the zinc-coordinated aryloxide ligand. For this family of zinc aryloxide complexes, a correlation was discovered between the chemical shift position of the N(3)-H proton resonance and the pK(a) of the parent aryl alcohol. This correlation indicates that the strength of the hydrogen bonding interaction involving the zinc-bound aryloxide oxygen is increasing as the aryloxide moiety increases in basicity. PMID- 12079476 TI - Redox energetics and kinetics of uranyl coordination complexes in aqueous solution. AB - Detailed voltammetric results for five uranyl coordination complexes are presented and analyzed using digital simulations of the voltammetric data to extract thermodynamic (E(1/2)) and heterogeneous electron-transfer kinetic (k(0) and alpha) parameters for the one-electron reduction of UO(2)(2+) to UO(2)(+). The complexes and their corresponding electrochemical parameters are the following: [UO(2)(OH(2))(5)](2+) (E(1/2) = -0.169 V vs Ag/AgCl, k(0) = 9.0 x 10( 3) cm/s, and alpha = 0.50); [UO(2)(OH)(5)](3-) (-0.927 V, 2.8 x 10(-3) cm/s, 0.46); [UO(2)(C(2)H(3)O(2))(3)](-) (-0.396 V, approximately 0.1 cm/s, approximately 0.5); [UO(2)(CO(3))(3)](4-) (-0.820 V, 2.6 x 10(-5) cm/s, 0.41); [UO(2)Cl(4)](2-) (-0.065 V, 9.2 x 10(-3) cm/s, 0.30). Differences in the E(1/2) values are attributable principally to differences in the basicity of the equatorial ligands. Differences in rate constants are considered within the context of Marcus theory of electron transfer, but no specific structural change(s) in the complexes between the two oxidation states can be uniquely identified with the underlying variability in the heterogeneous rate constants and electron-transfer coefficients. PMID- 12079477 TI - Bimetallic zirconium heterocycles supported by boron-oxygen ligands. AB - Reaction between bis(cyclopentadienyl)dimethylzirconium, ZrCp(2)(CH(3))(2), and phenylboronic anhydride, (PhBO)(3), resulted in the formation of the heterocyclic dimer [ZrCp(2)[mu-O(2)BPh]](2) (1); no reaction was observed with the mesityl derivative, (mesBO)(3). Compound 1 was also synthesized from the protonolysis reaction between ZrCp(2)(CH(3))(2) and in situ generated phenylboronic acid, PhB(OH)(2). This approach was extended to afford the analogous complexes [ZrCp(2)[mu-O(2)BAr]](2) (2, Ar = mes; 3, Ar = C(6)F(5)) from the corresponding isolable arylboronic acids, ArB(OH)(2). The molecular structures of 1-3, determined by X-ray diffraction techniques, revealed a common, dimeric motif consisting of a central "Zr(2)B(2)O(4)" metallacycle. Variations in the bond parameters within the heterocycle are attributed to the differing steric and electronic properties of the aryl substituents at boron. PMID- 12079478 TI - Reactive intermediates relevant to the carbonylation of CH(3)Mn(CO)(5). Activation parameters for key dynamic processes. AB - Reported is a time-resolved infrared and optical kinetics investigation of the transient species CH(3)C(O)Mn(CO)(4) (I(Mn)) generated by flash photolysis of the acetyl manganese pentacarbonyl complex CH(3)C(O)Mn(CO)(5) (A(Mn)) in cyclohexane and in tetrahydrofuran. Activation parameters were determined for CO trapping of I(Mn) to regenerate A(Mn) (rate = k(CO) [CO][I(Mn)]) as well as the methyl migration pathway to form methylmanganese pentacarbonyl CH(3)Mn(CO)(5) (M(Mn)) (rate = k(M)[I(Mn)]). These values were Delta H(++)(CO) = 31 +/- 1 kJ mol(-1), Delta S(++)(CO) = -64 +/- 3 J mol(-1) K(-1), Delta H(++)(M) = 35 +/- 1 kJ mol( 1), and Delta S(++)(M) = -111 +/- 3 J mol(-1) K(-1). Substantially different activation parameters were found for the methyl migration kinetics of I(Mn) in THF solutions where Delta H(++)(M) = 68 +/- 4 kJ mol(-1) and Delta S(++)(M) = 10 +/- 10 J mol(-1) K(-1), consistent with the earlier conclusion (Boese, W. T.; Ford, P. C. J. Am. Chem. Soc. 1995, 117, 8381-8391) that the composition of I(Mn) is different in these two media. The possible isotope effect on k(M) was also evaluated by studying the intermediates generated from flash photolysis of CD(3)C(O)Mn(CO)(5) in cyclohexane, but this was found to be nearly negligible (k(M)(h)/k(M)(d) (298 K) = 0.97 +/- 0.05, Delta H(++)(M)(d) = 37 +/- 4 kJ mol( 1), and Delta S(++)(M)(d) = -104 +/- 12 J mol(-1) K(-1)). The relevance to the migratory insertion mechanism of CH(3)Mn(CO)(5), a model for catalytic carbonylations, is discussed. PMID- 12079479 TI - Metal Cyanide Ions Mx(CN)y]+,- in the gas phase: M = Fe, Co, Ni, Zn, Cd, Hg, Fe + Ag, Co + Ag. AB - The generation of metal cyanide ions in the gas phase by laser ablation of M(CN)(2) (M = Co, Ni, Zn, Cd, Hg), Fe(III)[Fe(III)(CN)(6)] x xH(2)O, Ag(3)[M(CN)(6)] (M = Fe, Co), and Ag(2)[Fe(CN)(5)(NO)] has been investigated using Fourier transform ion cyclotron resonance mass spectrometry. Irradiation of Zn(CN)(2) and Cd(CN)(2) produced extensive series of anions, [Zn(n)(CN)(2n+1)](-) (1 < or = n < or = 27) and [Cd(n)(CN)(2n+1)](-) (n = 1, 2, 8-27, and possibly 29, 30). Cations Hg(CN)(+) and [Hg(2)(CN)(x)](+) (x = 1-3), and anions [Hg(CN)(x)](-) (x = 2, 3), are produced from Hg(CN)(2). Irradiation of Fe(III)[Fe(III)(CN)(6)] x xH(2)O gives the anions [Fe(CN)(2)](-), [Fe(CN)(3)](-), [Fe(2)(CN)(3)](-), [Fe(2)(CN)(4)](-), and [Fe(2)(CN)(5)](-). When Ag(3)[Fe(CN)(6)] is ablated, [AgFe(CN)(4)](-) and [Ag(2)Fe(CN)(5)](-) are observed together with homoleptic anions of Fe and Ag. The additional heterometallic complexes [AgFe(2)(CN)(6)](-), [AgFe(3)(CN)(8)](-), [Ag(2)Fe(2)(CN)(7)](-), and [Ag(3)Fe(CN)(6)](-) are observed on ablation of Ag(2)[Fe(CN)(5)(NO)]. Homoleptic anions [Co(n)(CN)(n+1)](-) (n = 1 3), [Co(n)(CN)(n+2)](-) (n = 1-3), [Co(2)(CN)(4)](-), and [Co(3)(CN)(5)](-) are formed when anhydrous Co(CN)(2) is the target. Ablation of Ag(3)[Co(CN)(6)] yields cations [Ag(n)(CN)(n-1)](+) (n = 1-4) and [Ag(n)Co(CN)(n)](+) (n = 1, 2) and anions [Ag(n)(CN)(n+1)](-) (n = 1-3), [Co(n)(CN)(n-1)](-) (n = 1, 2), [Ag(n)Co(CN)(n+2)](-) (n = 1, 2), and [Ag(n)Co(CN)(n+3)](-) (n = 0-2). The Ni(I) species [Ni(n)(CN)(n-1)](+) (n = 1-4) and [Ni(n)(CN)(n+1)](-) (n = 1-3) are produced when anhydrous Ni(CN)(2) is irradiated. In all cases, CN(-) and polyatomic carbon nitride ions C(x)N(y)(-) are formed concurrently. On the basis of density functional calculations, probable structures are proposed for most of the newly observed species. General structural features are low coordination numbers, regular trigonal coordination stereochemistry for d(10) metals but distorted trigonal stereochemistry for transition metals, the occurrence of M-CN M and M(-CN-)(2)M bridges, addition of AgCN to terminal CN ligands, and the occurrence of high spin ground states for linear [M(n)(CN)(n+1)](-) complexes of Co and Ni. PMID- 12079480 TI - Spin dimer analysis of the spin exchange interactions in paramelaconite Cu(4)O(3) and its analogue Ag(2)Cu(2)O(3) and the spin ordering of the Cu(2)O(3) spin lattice leading to their magnetic phase transitions. AB - The magnetic structures of the Cu(2)O(3) spin lattices present in Cu(4)O(3) and Ag(2)Cu(2)O(3) were analyzed by studying their spin exchange interactions on the basis of spin dimer analysis. Calculations of spin exchange parameters were calibrated by studying LiCuVO(4) whose intrachain and interchain antiferromagnetic spin exchange parameters are known experimentally. The magnetic phase transition of Cu(4)O(3) at 42.3 K doubles the unit cell along each crystallographic direction. The spin arrangements of the Cu(2)O(3) lattice consistent with this experimental observation are different from conventional antiferromagnetic ordering. Our analysis indicates that spin fluctuation should occur in Cu(4)O(3), low-dimensional magnetism should be more important than magnetic frustration in Cu(4)O(3), and Ag(2)Cu(2)O(3) and Cu(4)O(3) should have similar structural and magnetic properties. PMID- 12079481 TI - Photodriven electron and energy transfer from a light-harvesting metallodendrimer. AB - Intermolecular electron and energy transfer from a light-harvesting metallodendrimer [Ru[bpy(C-450)(4)](3)](2+), where bpy(C-450)(4) is a 2,2' bipyridine derivative containing 4 coumarin-450 units connected together through aryl ether linkages, is observed in acetonitrile solutions at room temperature. The model complex [Ru(dmb)(3)](2+), where dmb is 4,4'-dimethyl-2,2'-bipyridine, is included for quantitative comparison. The excited states of both compounds are metal-to-ligand charge transfer in nature and participate in excited-state electron and triplet energy transfer processes. Quenching constants were determined from luminescence and time-resolved absorption experiments at constant ionic strength. [Ru[bpy(C-450)(4)](3)](2+) displays significantly slower quenching rates to molecular oxygen and methyl viologen relative to the other processes investigated. Triplet energy transfer from [Ru[bpy(C-450)(4)](3)](2+) to 9-methylanthracene is quantitatively indistinguishable from [Ru(dmb)(3)](2+) while reductive electron transfer from phenothiazine was slightly faster in the former. With the exception of dioxygen quenching, our results indicate that the current dendritic structure is ineffective in shielding the core from bimolecular electron and triplet energy transfer reactions. Electrochemical measurements of [Ru[bpy(C-450)(4)](3)](2+) reveal irreversible oxidative processes at potentials slightly negative to the Ru(III/II) potential that are assigned to oxidations in the dendritic structure. Excited-state oxidative electron-transfer reactions facilitate this process resulting in the reduction of ground-state Ru(III) to Ru(II) and the trapping of the methyl viologen radical cation (MV(*+)) when methyl viologen serves as the quencher. This process generates a minimum of 9 MV(*+)'s for every [Ru[bpy(C-450)(4)](3)](2+) molecule and disassembles the metallodendrimer, resulting in the production of a [Ru(dmb)(3)](2+)-like species and "free" C-450-like dyes. PMID- 12079482 TI - Challenges for the translation of T cell costimulatory blockade therapies to the clinic. PMID- 12079483 TI - Spontaneous regression of neoplasms: new possibilities for immunotherapy. AB - In mammalian cells, neoplastic transformation is directly associated with the expression of oncogenes, loss or simple inactivation of the function of tumour suppressor genes and the production of certain growth factors. Genes for suppression of the development of the neoplastic cellular immunophenotype, as well as inhibitory growth factors, have regulatory functions within the normal processes of cell division and differentiation. Telomerase (a ribonucleoprotein polymerase) activation is frequently detected in various neoplasms. Telomerase activation is regarded as essential for cell immortalisation and its inhibition may result in spontaneous regression of neoplasms. This phenomenon of neoplasms occurs when the malignant tissue mass partially or completely disappears without any treatment or as a result of a therapy considered inadequate to influence systemic neoplastic growth. This definition makes it clear that the term 'spontaneous regression' applies to neoplasms in which the overall malignant disease is not necessarily cured and to cases where the regression may not be complete or permanent. A number of possible mechanisms of spontaneous regression are reviewed, with the understanding that no single mechanism can completely account for this phenomenon. The application of the newest immunological, molecular biological and genetic insights for more individualised and adequate antineoplastic immunotherapy (alternative biotherapy) is also discussed. PMID- 12079484 TI - Angiogenesis inhibitors and radiotherapy of primary tumours. AB - The modern approach to cancer therapy involves combinations of surgery, radiation and drugs targeted against malignant tumour cells. However, expanding knowledge in the fields of angiogenesis and vascular biology over the last several years has led to the investigation of therapeutic strategies targeted to the vasculature (proliferating and non-proliferating endothelial cells) in combination with standard therapy. It is the objective of this review to describe the potential use of antiangiogenic therapy, targeted to the proliferating endothelium, from the point of view of the radiation oncologist. This review will describe the concept of a two-cell compartment model for tumours, with both the endothelial cells as well as the tumour cells being potential targets for radiotherapy. This review will then explore the promising evidence and rationale for combining antiangiogenic drugs and radiotherapy to enhance local control. PMID- 12079485 TI - New developments in immunosuppressive therapy in renal transplantation. AB - The introduction of new immunosuppressive agents and protocols has improved outcomes for renal transplant recipients by decreasing the risk of rejection and by increasing the function and lifespan of the allograft. This article reviews the major changes in the combinations of therapies used: calcineurin inhibitors, target of rapamycin inhibitors, mycophenolate mofetil, non-depleting monoclonal versus depleting monoclonal and polyclonal antibodies for induction and increasing emphasis on protocols for reduction or avoidance of steroids and calcineurin inhibitors. The new agents with novel immunological targets such as anti-CD40 ligand, LEA29Y, FTY720, anti-CD20 (rituximab, Rituxan, Mabthera) and anti-CH52 (alemtuzumab, Campath), which are under development but have yet to survive the rigors of clinical trials are also discussed. In the presence of low early rejection rates, immunosuppressive therapy is setting new goals such as better graft function (glomerular filtration rates), reduction in adverse effects such as hypertension, hyperlipidaemia and drug toxicity and, above all, the prevention of late graft deterioration. PMID- 12079486 TI - Immunoisolation techniques for islet cell transplantation. AB - Diabetes remains a devastating disease, with tremendous cost in terms of human suffering and healthcare expenditures. The burden of diabetes is primarily related to the multiple complications, including retinopathy, nephropathy, neuropathy and cardiovascular disease that can develop as the disease progresses. It has been shown that these complications can be prevented, and in some cases, reversed by islet cell transplantation, which, until recently, had remained elusive as a viable routine treatment modality. In recent studies, islet cell transplantation has shown great promise as a viable alternative to solid pancreas transplantation. However, severe shortage of human pancreases and the need to use immunosuppressive drugs to prevent transplant rejection, remain major obstacles to routine use of islet cell transplants for the treatment of patients with Type 1 diabetes. In the attempt to overcome these barriers, many procedures have been designed to immunoisolate islet cells for transplantation. The ultimate goal in islet cell transplantation is the availability of unlimited supply of cells to be transplanted in a simple procedure performed with little or no use of immunosuppressive drugs. The development of reliable procedures to immunoisolate islets by microencapsulation prior to transplantation has a great deal of potential to accomplish this objective. PMID- 12079487 TI - IL-12/IL-2 combination cytokine therapy for solid tumours: translation from bench to bedside. AB - A broad range of approaches are under active investigation for the biological therapy of cancer, in particular, strategies directed at host immune response potentiation. These efforts have been fuelled by studies demonstrating the presence of an endogenous, but ineffective, host antitumour immune response and a greater understanding of the key factors which regulate this response. These mechanisms involve complex interactions between various effector cell populations, soluble factors and the tumour itself and are determined by the timing and relative intensity of positive and negative autoregulatory pathways, as well as a variety of immunosuppressive effects capable of mediating tumour self-defence. Based on these observations, immunotherapeutic regimens have been developed to potentiate antigen-specific sensitisation of effector cells with tumour vaccines/adjuvants, expand and amplify the number and function of effector cells, and to counteract suppressive pathways engaged by tumour cells themselves. Significant effort has focused on evaluating the use of exogenous cytokines, administered either systemically or locally into the tumour site via gene therapy. Several cytokines have demonstrated unique activity in the preclinical setting, including IL-2 and IFN-alpha -inducing cytokines such as IL12 and IL18. Most notably, later studies have now attempted to build on the clinical efficacy of IL-2 alone, to define combinations of agents with synergistic immunoregulatory and/or antitumour efficacy. Several lines of evidence suggest that IL-12 and IL-2 provide complementary immunoregulatory signals and have now shown that in combination, these two cytokines mediate synergistic antitumour activity in preclinical tumour models. This paper will review existing data regarding mechanisms of interaction between IL-2 and IL-12 in vitro and in preclinial models and describe future opportunities for the investigation of these potentially promising cytokines in the treatment of cancer. PMID- 12079488 TI - Ecdysone-controlled expression of transgenes. AB - Inducible expression systems show great potential for use in human gene therapy and systems based on insect ecdysone receptors are particularly promising candidates. This article describes such systems and reviews actual and potential uses of ecdysone-controlled transgenes in vitro and in vivo. The ligand specificity of ecdysone receptor-based systems is considered, along with the safety and efficacy of the ecdysteroid and non-steroidal compounds used to activate them. PMID- 12079489 TI - Gene therapy for retinal and choroidal diseases. AB - The eye is a small compartment separated from the systemic circulation by the blood-ocular barriers, providing advantages for intraocular gene transfer - an approach which is being investigated for several types of retinal and choroidal diseases. A compelling application is gene replacement for homozygous loss-of function mutations in genes differentially expressed in photoreceptors or retinal pigmented epithelial (RPE) cells that result in retinal degeneration. Considerable progress has been made in this area, including demonstration of return of visual function in RPE65 (-/-) dogs after subretinal injection of adeno associated viral vectors encoding RPE65, providing groundwork for a clinical trial in patients with Leber's Congenital Amaurosis. Proof of principle has been provided for intraocular gene transfer of ribozymes for dominantly inherited retinal degenerations. Survival factor gene therapy shows promise for treatments that may be used in multiple retinal degenerations. Transduction of intraocular and/or periocular cells with constructs that encode antiangiogenic proteins provides a new approach for sustained local delivery treatment of retinal and choroidal neovascularisation. While considerable investigation remains to work out critical details, there is substantial evidence suggesting that in the near future, gene therapy-based treatments will be an important addition to what is currently offered to patients with retinal and/or choroidal diseases. PMID- 12079491 TI - Microarray technology and gene expression analysis for the study of angiogenesis. AB - Angiogenesis plays a major role in multiple disease processes including cancer, and new agents that modulate angiogenesis are rapidly entering clinical trials. The understanding of the biological mechanisms and downstream effects for many of these agents is poorly understood. It is therefore important that methods evolve to understand how an agent regulates angiogenesis, in order to promote a higher percentage of successful drug candidates. With the emergence of microarray technology for the evaluation of gene expression, researchers have a powerful tool for dissecting the biological mechanisms of angiogenesis. However, huge data sets and complex statistics pose a hurdle for the investigator to obtain useful and meaningful data. To eliminate problems in data analysis, proper design and planning prior to performing a microarray experiment is crucial to making valid conclusions. This review will discuss the critical factors in designing, performing and analysing microarray experiments, and the utility of various models of angiogenesis for microarray analysis. PMID- 12079490 TI - The role of quantitative PCR for the immune monitoring of cancer patients. AB - Human tumour immunology is at a standstill whereas implemented cancer vaccines have shown effectiveness in inducing immune responses detectable in circulating lymphocytes. In most circumstances, however, such immune responses are not sufficient to induce cancer regression. This paradoxical observation could be explained in several ways depending upon the immunological endpoint used for immune monitoring. For instance, analysis of immune responses in circulating lymphocytes that address the presence of T cells bearing T-cell receptors specific for the epitope used for vaccination, can accurately enumerate the number of T cells elicited by the vaccines but does not yield information about their functional status. Other monitoring strategies may yield general information about the reactivity of various T cells in response to a relevant stimulus and, therefore, may provide information more relevant to the purpose of the immunisation. Furthermore, the material used to monitor immune responses may, in itself, determine the significance of the findings obtained. In the assessment of the therapeutic efficacy of specific cancer treatment, analysis of immune responses in circulating lymphocytes (systemic response) may not be as relevant as the analysis of the same effector populations within the tumour microenvironment (peripheral response). This review will describe a novel approach that allows extreme flexibility in the analysis of systemic and peripheral responses by accurately measuring the level of expression of relevant genes using quantitative real-time reverse transcriptase polymerase chain reaction. PMID- 12079492 TI - 3rd international symposium on genetic anticancer agents. AB - The 3rd International Symposium on Genetic Anticancer Agents was an initiative of NDDO Research Foundation, which is an internationally operating organisation exclusively devoted to the research and development of new strategies for the treatment of cancer. The meeting focused on the preclinical and clinical development of various genetic agents for cancer treatment. Leading world experts came to Amsterdam to present recent results of their highly advanced research and to discuss novel approaches to cancer treatment. In addition to the scientific sessions, the organisers prepared an educational session devoted to the development process of a gene therapy product and to the design of clinical trials. The educational session aroused a great interest from the participants. The meeting highlights from different scientific sessions are presented. PMID- 12079493 TI - Stopped-flow kinetic studies of electron transfer in the reductase domain of neuronal nitric oxide synthase: re-evaluation of the kinetic mechanism reveals new enzyme intermediates and variation with cytochrome P450 reductase. AB - The reduction by NADPH of the FAD and FMN redox centres in the isolated flavin reductase domain of calmodulin-bound rat neuronal nitric oxide synthase (nNOS) has been studied by anaerobic stopped-flow spectroscopy using absorption and fluorescence detection. We show by global analysis of time-dependent photodiode array spectra, single wavelength absorption and NADPH fluorescence studies, that at least four resolvable steps are observed in stopped-flow studies with NADPH and that flavin reduction is reversible. The first reductive step represents the rapid formation of an equilibrium between an NADPH-enzyme charge-transfer species and two-electron-reduced enzyme bound to NADP(+). The second and third steps represent further reduction of the enzyme flavins and NADP(+) release. The fourth step is attributed to the slow accumulation of an enzyme species that is inferred not to be relevant catalytically in steady-state reactions. Stopped-flow flavin fluorescence studies indicate the presence of slow kinetic phases, the timescales of which correspond to the slow phase observed in absorption and NADPH fluorescence transients. By analogy with stopped-flow studies of cytochrome P450 reductase, we attribute these slow fluorescence and absorption changes to enzyme disproportionation and/or conformational change. Unlike for the functionally related cytochrome P450 reductase, transfer of the first hydride equivalent from NADPH to nNOS reductase does not generate the flavin di-semiquinoid state. This indicates that internal electron transfer is relatively slow and is probably gated by NADP(+) release. Release of calmodulin from the nNOS reductase does not affect the kinetics of inter-flavin electron transfer under stopped-flow conditions, although the observed rate of formation of the equilibrium between the NADPH-oxidized enzyme charge-transfer species and two-electron-reduced enzyme bound to NADP(+) is modestly slower in calmodulin-depleted enzyme. Our studies indicate the need for significant re-interpretation of published kinetic data for electron transfer in the reductase domain of neuronal nitric oxide synthase. PMID- 12079494 TI - Calmodulin controls organization of the actin cytoskeleton via regulation of phosphatidylinositol (4,5)-bisphosphate synthesis in Saccharomyces cerevisiae. AB - Phosphoinositides regulate a wide range of cellular processes, including proliferation, survival, cytoskeleton remodelling and membrane trafficking, yet the mechanisms controlling the kinases, phosphatases and lipases that modulate phosphoinositide levels are poorly understood. In the present study, we describe a mechanism controlling MSS4, the sole phosphatidylinositol (4)-phosphate 5 kinase in Saccharomyces cerevisiae. Mutations in MSS4 and CMD1, encoding the small Ca(2+)-binding protein calmodulin, confer similar phenotypes, including loss of viability and defects in endocytosis and in organization of the actin cytoskeleton. Overexpression of MSS4 suppresses the growth and actin defects of cmd1-226, a temperature-sensitive calmodulin mutant which is defective in the organization of the actin cytoskeleton. Finally, the cmd1-226 mutant exhibits reduced levels of phosphatidylinositol (4,5)-bisphosphate. These findings suggest that calmodulin positively controls MSS4 activity and thereby the actin cytoskeleton. PMID- 12079495 TI - Distinct sites for myotoxic and membrane-damaging activities in the C-terminal region of a Lys49-phospholipase A2. AB - Bothropstoxin-I (BthTx-I) is a Lys(49)-phospholipase A(2) from the venom of Bothrops jararacussu which demonstrates both myotoxic and Ca(2+)-independent membrane-damaging activities. The structural determinants of these activities are poorly defined, therefore site-directed mutagenesis has been used to substitute all cationic and aromatic residues between positions 115 and 129 in the C terminal loop region of the protein. Substitution of lysine and arginine residues with alanine in the region 117-122 resulted in a significant reduction of myotoxic activity of the recombinant BthTx-I. With the exception of Lys(122), these same substitutions did not significantly alter the Ca(2+)-independent membrane-damaging activity. In contrast, substitution of the positively-charged residues at positions 115, 116 and 122 resulted in reduced Ca(2+)-independent membrane-damaging activity but, with the exception of Lys(122), had no effect on myotoxicity. These results indicate that the two activities are independent and are determined by discrete yet partially overlapping motifs in the C-terminal loop. Results from site-directed mutagenesis of the aromatic residues in the same part of the protein suggest that a region including residues 115-119 interacts superficially with the membrane interface and that the residues around position 125 partially insert into the lipid membrane. These results represent the first detailed mapping of a myotoxic site in a phospholipase A(2), and support a model of a Ca(2+)-independent membrane-damaging mechanism in which the C-terminal region of BthTx-I interacts with and contributes to the perturbation of the phospholipid bilayer. PMID- 12079496 TI - The effects of weather on fungal abundance and richness among 25 communities in the Intermountain West. AB - BACKGROUND: Because moisture and temperature influence the growth of fungi, characterizing weather conditions favorable for fungi may be used to predict the abundance and richness of fungi in habitats with different climate conditions. To estimate habitat favorability to fungi, we examined the relationship of fungal abundance and species richness to various weather and environmental parameters in the Intermountain West. We cultured fungi from air and leaf surfaces, and collected continuous temperature and relative humidity measures over the growing season at 25 sites. RESULTS: Fungal richness was positively correlated with fungal abundance (r = 0.75). Measures of moisture availability, such as relative humidity and vapor pressure deficit, explained more of the variance in fungal abundance and richness than did temperature. Climate measurements from nearby weather stations were good predictors of fungal abundance and richness but not as good as weather measurements obtained in the field. Weather variables that took into account the proportion of time habitats experienced favorable or unfavorable relative humidity and temperatures were the best predictors, explaining up to 56% of the variation in fungal abundance and 72% for fungal richness. CONCLUSION: Our results suggest that the abundance and richness of fungi in a habitat is limited by the duration of unfavorable weather conditions. Because fungal pathogens likely have similar abiotic requirements for growth as other fungi, characterizing weather conditions favorable for fungi also may be used to predict the selective pressures imposed by pathogenic fungi on plants in different habitats. PMID- 12079497 TI - Embryonic stem cells and mice expressing different GFP variants for multiple non invasive reporter usage within a single animal. AB - BACKGROUND: Non-invasive autofluorescent reporters have revolutionized lineage labeling in an array of different organisms. In recent years green fluorescent protein (GFP) from the bioluminescent jellyfish Aequoria Victoria has gained popularity in mouse transgenic and gene targeting regimes 1. It offers several advantages over conventional gene-based reporters, such as lacZ and alkaline phosphatase, in that its visualization does not require a chromogenic substrate and can be realized in vivo. We have previously demonstrated the utility and developmental neutrality of enhanced green fluorescent protein (EGFP) in embryonic stem (ES) cells and mice 2. RESULTS: In this study we have used embryonic stem (ES) cell-mediated transgenesis to test the enhanced cyan fluorescent protein (ECFP) and enhanced yellow fluorescent protein (EYFP), two mutant and spectrally distinct color variants of wild type (wt) GFP. We have also tested DsRed1, the novel red fluorescent protein reporter recently cloned from the Discostoma coral by virtue of its homology to GFP. To this end, we have established lines of ES cells together with viable and fertile mice having widespread expression of either the ECFP or EYFP GFP-variant reporters. However, we were unable to generate equivalent DsRed1 lines, suggesting that DsRed1 is not developmentally neutral or that transgene expression cannot be sustained constitutively. Balanced (diploid <-> diploid) and polarized (tetraploid <-> diploid) chimeras comprising combinations of the ECFP and EYFP ES cells and/or embryos, demonstrate that populations of cells expressing each individual reporter can be distinguished within a single animal. CONCLUSIONS: GFP variant reporters are unique in allowing non-invasive multi-spectral visualization in live samples. The ECFP and EYFP-expressing transgenic ES cells and mice that we have generated provide sources of cells and tissues for combinatorial, double tagged recombination experiments, chimeras or transplantations. PMID- 12079498 TI - Low solubility of unconjugated bilirubin in dimethylsulfoxide--water systems: implications for pKa determinations. AB - BACKGROUND: Aqueous pKa values of unconjugated bilirubin are important determinants of its solubility and transport. Published pKa data on an analog, mesobilirubin-XIIIalpha, studied by 13C-NMR in buffered solutions containing 27 and 64 vol% (C2H3)2SO because of low aqueous solubility of mesobilirubin, were extrapolated to obtain pKa values in water of 4.2 and 4.9. Previous chloroform water partition data on bilirubin diacid led to higher estimates of its pKa, 8.12 and 8.44, and its aqueous solubility. A thermodynamic analysis, using this solubility and a published solubility in DMSO, suggested that the systems used to measure 13C-NMR shifts were highly supersaturated. This expectation was assessed by measuring the residual concentrations of bilirubin in the supernatants of comparable DMSO-buffer systems, after mild centrifugation to remove microprecipitates. RESULTS: Extensive sedimentation was observed from numerous systems, many of which appeared optically clear. The very low supernatant concentrations at the lowest pH values (4.1-5.9) were compatible with the above thermodynamic analysis. Extensive sedimentation and low supernatant concentrations occurred also at pH as high as 7.2. CONCLUSIONS: The present study strongly supports the validity of the aqueous solubility of bilirubin diacid derived from partition data, and, therefore, the corresponding high pKa values. Many of the mesobilirubin systems in the 13C-NMR studies were probably supersaturated, contained microsuspensions, and were not true solutions. This, and previously documented errors in pH determinations that caused serious errors in pKa values of the many soluble reference acids and mesobilirubin, raise doubts regarding the low pKa estimates for mesobilirubin from the 13C-NMR studies. PMID- 12079499 TI - Functional group interactions of a 5-HT3R antagonist. AB - BACKGROUND: Lerisetron, a competitive serotonin type 3 receptor (5-HT3R) antagonist, contains five functional groups capable of interacting with amino acids in the 5-HT3R binding site. Site directed mutagenesis studies of the 5 HT3AR have revealed several amino acids that are thought to form part of the binding domain of this receptor. The specific functional groups on the ligand that interact with these amino acids are, however, unknown. Using synthetic analogs of lerisetron as molecular probes in combination with site directed mutagenesis, we have identified some of these interactions and have proposed a model of the lerisetron binding site. RESULTS: Two analogs of lerisetron were synthesized to probe 5-HT3R functional group interactions with this compound. Analog 1 lacks the N1 benzyl group of lerisetron and analog 2 contains oxygen in place of the distal piperazine nitrogen. Both analogs show significantly decreased binding affinity to wildtype 5-HT3ASRs. Mutations at W89, R91, Y142 and Y152 produced significant decreases in binding compared to wildtype receptors. Binding affinities of analogs 1 and 2 were altered only by mutations at W89, and Y152. CONCLUSIONS: Based on the data obtained for lerisetron and analogs 1 and 2, we have proposed a tentative model of the lerisetron binding pocket of the 5 HT3ASR. According to this model, The N-benzyl group interacts in a weak interaction with R91 while the benzimidazole group interacts with W89. Our data support an interaction of the distal amino nitrogen with Y142 and Y152. PMID- 12079500 TI - Identification of critical residues in loop E in the 5-HT3ASR binding site. AB - BACKGROUND: The serotonin type 3 receptor (5-HT3R) is a member of a superfamily of ligand gated ion channels. All members of this family share a large degree of sequence homology and presumably significant structural similarity. A large number of studies have explored the structure-function relationships of members of this family, particularly the nicotinic and GABA receptors. This information can be utilized to gain additional insights into specific structural and functional features of other receptors in this family. RESULTS: Thirteen amino acids in the mouse 5-HT3ASR that correspond to the putative E binding loop of the nicotinic alpha7 receptor were chosen for mutagenesis. Due to the presence of a highly conserved glycine in this region, it has been suggested that this binding loop is comprised of a hairpin turn and may form a portion of the ligand-binding site in this ion channel family. Mutation of the conserved glycine (G147) to alanine eliminated binding of the 5-HT3R antagonist [3H]granisetron. Three tyrosine residues (Y140, Y142 and Y152) also significantly altered the binding of 5-HT3R ligands. Mutations in neighboring residues had little or no effect on binding of these ligands to the 5-HT3ASR. CONCLUSION: Our data supports a role for the putative E-loop region of the 5-HT3R in the binding of 5-HT, mCPBG, d-tc and lerisetron. 5-HT and mCPBG interact with Y142, d-tc with Y140 and lerisetron with both Y142 and Y152. Our data also provides support for the hypothesis that this region of the receptor is present in a loop structure. PMID- 12079501 TI - Enzyme replacement reverses abnormal cerebrovascular responses in Fabry disease. AB - BACKGROUND: Fabry disease is a lysosomal X-linked enzyme deficiency of alpha galactosidase A associated with an increased mortality and morbidity due to renal failure, cardiac disease and early onset stroke. METHODS: We examined the functional blood flow response of the brain after visual stimulation (reversing checkerboard pattern), and cerebral vasoreactivity following acetazolamide (15 mg/kg) with [15O]H2O and positron emission tomography (PET) in Fabry disease. Twenty-six hemizygous patients (age range 19-47 years) were enrolled in a randomized double-blind placebo-controlled 6-month trial of enzyme replacement therapy administered by intravenous infusion every two weeks. Regional cerebral blood flow (rCBF) was measured with PET at the beginning and end of the trial. RESULTS: Fabry patients had a significantly greater increase in rCBF following visual stimulation and acetazolamide challenge compared to controls. Visual reactivity was normal. The time for recovery of the cerebral vasculature following acetazolamide was prolonged in Fabry patients compared to controls. The abnormal rCBF response induced by visual stimulation and acetazolamide decreased significantly following enzyme replacement therapy, as did the prolonged recovery of the cerebral vasculature. CONCLUSIONS: Enzyme replacement therapy reverses the exaggerated cerebrovascular response in Fabry disease. PMID- 12079502 TI - House dust mite barrier bedding for childhood asthma: randomised placebo controlled trial in primary care [ISRCTN63308372]. AB - BACKGROUND: The house dust mite is the most important environmental allergen implicated in the aetiology of childhood asthma in the UK. Dust mite barrier bedding is relatively inexpensive, convenient to use, and of proven effectiveness in reducing mattress house dust mite load, but no studies have evaluated its clinical effectiveness in the control of childhood asthma when dispensed in primary care. We therefore aimed to evaluate the effectiveness of house dust mite barrier bedding in children with asthma treated in primary care. METHODS: Pragmatic, randomised, double-blind, placebo controlled trial conducted in eight family practices in England. Forty-seven children aged 5 to 14 years with confirmed house dust mite sensitive asthma were randomised to receive six months treatment with either house dust mite barrier or placebo bedding. Peak expiratory flow was the main outcome measure of interest; secondary outcome measures included asthma symptom scores and asthma medication usage. RESULTS: No difference was noted in mean monthly peak expiratory flow, asthma symptom score, medication usage or asthma consultations, between children who received active bedding and those who received placebo bedding. CONCLUSIONS: Treating house dust mite sensitive asthmatic children in primary care with house dust mite barrier bedding for six months failed to improve peak expiratory flow. Results strongly suggest that the intervention made no impact upon other clinical features of asthma. PMID- 12079503 TI - Cellophane based mini-prep method for DNA extraction from the filamentous fungus Trichoderma reesei. AB - BACKGROUND: Methods for the extraction of DNA from filamentous fungi are frequently laborious and time consuming because most of the available protocols include maceration in liquid nitrogen after the mycelium has been grown in a liquid culture. This paper describes a new method to replace those steps, which involves the growth of the mycelium on cellophane disks overlaid on solid medium and the use of glass beads for cell wall disruption. RESULTS: Extractions carried out by this method provided approximately 2 microg of total DNA per cellophane disk for the filamentous fungus Trichoderma reesei. To assess the DNA's quality, we made a PCR (Polymerase Chain Reaction) amplification of a gene introduced by a transformation in this fungus's genome (hph gene), with successful results. We also confirmed the quality of the DNA by the use of Southern blotting to analyze the presence of the same gene, which was easily detected, resulting in a sharply defined and strong band. CONCLUSIONS: The use of this method enabled us to obtain pure DNA from Trichoderma reesei, dispensing with the laborious and time consuming steps involved in most protocols. The DNA obtained was found to be suitable for PCR and Southern blot analyses. Another advantage of this method is the fact that several samples can be processed simultaneously, growing the fungus on multiple well cell culture plates. In addition, the absence of maceration also reduces sample handling, minimizing the risks of contamination, a particularly important factor in work involving PCR. PMID- 12079504 TI - Oxygen and nitrate-dependent regulation of dmsABC operon expression in Escherichia coli: sites for Fnr and NarL protein interactions. AB - BACKGROUND: Escherichia coli, can respire anaerobically using dimethyl sulfoxide (DMSO) or trimethylamine-N-oxide (TMAO) as the terminal electron acceptor for anaerobic energy generation. Expression of the dmsABC genes that encode the membrane-associated DMSO/TMAO reductase is positively regulated during anaerobic conditions by the Fnr protein and negatively regulated by the NarL protein when nitrate is present. RESULTS: The regions of dmsA regulatory DNA required for Fnr and NarL interactions in response to anaerobiosis and nitrate, respectively, were examined. Mutations within the Fnr site that deviated from the wild type sequence, TTGATaccgAACAA, or that removed an entire half-site, either impaired or abolished the anaerobic activation of dmsA-lacZ expression. The region for phosphorylated NarL (NarL-phosphate) binding at the dmsA promoter was identified by DNase I and hydroxyl radical footprinting methods. A large 97 bp region that overlaps the Fnr and RNA polymerase recognition sites was protected by NarL phosphate but not by the non-phosphorylated form of NarL. Hydroxyl radical footprinting analysis confirmed the NarL-phosphate DNase I protections of both dmsA strands and revealed 8-9 protected sites of 3-5 bp occurring at ten bp intervals that are offset by 3 bp in the 3' direction. CONCLUSION: These findings suggest that multiple molecules of phosphorylated NarL bind along one face of the DNA and may interfere with Fnr and/or RNA polymerase interactions at the dmsA regulatory region. The interplay of these transcription factors insures a hierarchical expression of the dmsABC genes when respiration of the preferred electron acceptors, oxygen and nitrate, is not possible. PMID- 12079505 TI - Transcriptional response of human mast cells stimulated via the Fc(epsilon)RI and identification of mast cells as a source of IL-11. AB - BACKGROUND: In asthma and other allergic disorders, the activation of mast cells by IgE and antigen induces the cells to release histamine and other mediators of inflammation, as well as to produce certain cytokines and chemokines. To search for new mast cell products, we used complementary DNA microarrays to analyze gene expression in human umbilical cord blood-derived mast cells stimulated via the high-affinity IgE receptor (Fc(epsilon)RI). RESULTS: One to two hours after Fc(epsilon)RI-dependent stimulation, more than 2,400 genes (about half of which are of unknown function) exhibited 2-200 fold changes in expression. The transcriptional program included changes in the expression of IL-11 and at least 30 other cytokines and chemokines. Human mast cells secreted 130-529 pg of IL 11/106 cells by 6 h after stimulation with anti-IgE. CONCLUSION: Our initial analysis of the transcriptional program induced in in vitro-derived human mast cells stimulated via the Fc(epsilon)RI has identified many products that heretofore have not been associated with this cell type, but which may significantly influence mast cell function in IgE-associated host responses. We also have demonstrated that mast cells stimulated via the Fc(epsilon)RI can secrete IL-11. Based on the previously reported biological effects of IL-11, our results suggest that production of IL-11 may represent one link between IgE dependent mast cell activation in subjects with allergic asthma and the development of a spectrum of structural changes in the airways of these individuals; such changes, collectively termed "airway remodeling," can constitute an important long term consequence of asthma. PMID- 12079506 TI - Negative regulation of mitochondrial VDAC channels by C-Raf kinase. AB - BACKGROUND: Growth of cancer cells results from the disturbance of positive and negative growth control mechanisms and the prolonged survival of these genetically altered cells due to the failure of cellular suicide programs. Genetic and biochemical approaches have identified Raf family serine/threonine kinases B-Raf and C-Raf as major mediators of cell survival. C-Raf cooperates with Bcl-2/Bcl-XL in suppression of apoptosis by a mechanism that involves targeting of C-Raf to the outer mitochondrial membrane and inactivation of the pro-apoptotic protein Bad. However, apoptosis suppression by C-Raf also occurs in cells lacking expression of Bad or Bcl-2. RESULTS: Here we show that even in the absence of Bcl-2/Bcl-XL, mitochondria-targeted C-Raf inhibits cytochrome c release and caspase activation induced by growth factor withdrawal. To clarify the mechanism of Bcl-2 independent survival control by C-Raf at the mitochondria a search for novel mitochondrial targets was undertaken that identified voltage dependent anion channel (VDAC), a mitochondrial protein (porin) involved in exchange of metabolites for oxidative phosphorylation. C-Raf forms a complex with VDAC in vivo and blocks reconstitution of VDAC channels in planar bilayer membranes in vitro. CONCLUSION: We propose that this interaction may be responsible for the Raf-induced inhibition of cytochrome c release from mitochondria in growth factor starved cells. Moreover, C-Raf kinase-induced VDAC inhibition may regulate the metabolic function of mitochondria and mediate the switch to aerobic glycolysis that is common to cancer cells. PMID- 12079507 TI - Promoter methylation of TSLC1 and tumor suppression by its gene product in human prostate cancer. AB - We recently identified TSLC1, a tumor suppressor gene in human lung cancer. Gene silencing by promoter methylation has been observed frequently in adenocarcinoma of the lung, liver, and pancreas. Here, we demonstrate that TSLC1 expression is also absent or markedly reduced in 3 of 4 prostate cancer cell lines. Promoter sequences of TSLC1 were heavily methylated in PPC-1 cells that lacked TSLC1 expression, supporting the idea that promoter methylation is strongly correlated with complete loss of gene expression. Promoter sequences of TSLC1 were also methylated significantly in 7 of 22 (32%) primary prostate cancers. Hypermethylation of the promoter occurred not only in advanced tumors, but also in relatively early-stage tumors. Restoration of TSLC1 expression substantially suppressed tumor formation of PPC-1 cells in nude mice. These findings indicate that alteration of TSLC1 is involved in prostate cancer. PMID- 12079508 TI - Trends in benign breast tumors in Japanese women, 1973-1995: experience of Hiroshima Tumor Tissue Registry. AB - Although some benign breast lesions such as multiple intraductal papilloma have been pointed out as a risk factor for breast cancer, there is little documentation about trends in the incidence of benign tumors of the breast in Japanese women. The author conducted an epidemiological study using data abstracted from the Hiroshima Tumor Tissue Registry file, which includes cases of malignant and benign neoplasms in Hiroshima prefecture between 1973 and 1995. A total of 17064 cases of female breast tumor were registered in the file between 1973 and 1995. The registration rate for fibroadenoma of the female breast was 19.5 among females and peaked during the 5-year period 1983 - 1987, while the fibroadenoma registration rate in Hiroshima gradually decreased between 1988 and 1995. The registration rate for intraductal papilloma has risen substantially in Hiroshima, with about a 5-fold increase among females between 1973 and 1995. The mean proportion of fibroadenoma cases accompanied by malignant tumors of the breast was 1.85%, and there was no significant change in the mean proportion between 1973 and 1995 (P = 0.17). On the other hand, the mean proportion of intraductal papilloma cases accompanied by malignant tumors of the breast gradually rose with about a 14-fold increase among females between 1973 and 1995. The significance of intraductal papilloma as a risk factor for breast cancer may have increased. PMID- 12079509 TI - Down-regulation of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx)-induced CYP1A2 expression is associated with bovine lactoferrin inhibition of MeIQx induced liver and colon carcinogenesis in rats. AB - The inhibitory influence of bovine lactoferrin (bLF) on induction of preneoplastic hepatic glutathione S-transferase placental form-positive (GST-P( +)) cell foci and colon aberrant crypt foci (ACF) by diethylnitrosamine (DEN) and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) was investigated in F344 rats. Rats were initially treated with DEN, then placed on basal diet containing MeIQx (200 ppm) alone, MeIQx plus 2% bLF, or MeIQx plus 0.2% bLF from week 2 to week 8, with partial hepatectomy performed at week 3. Concomitant administration of 2% or 0.2% bLF with MeIQx caused significant dose-dependent decreases in both number and unit area of GST-P(+) cell foci (2% bLF, P < 0.001; 0.2% bLF, P < 0.01). Similar results were observed for MeIQx-induced colon ACF in the groups without DEN treatment (2% and 0.2% bLF, P < 0.05). To investigate the underlying mechanisms, we analyzed the influence of bLF on levels of cytochrome P4501A2 (CYP1A2), a metabolically activating enzyme of MeIQx in the liver. The results demonstrated that combined administration of 2% bLF significantly reduced levels of MeIQx-induced CYP1A2 mRNA (P < 0.05) and protein (P < 0.05) to the normal levels, in association with reduced values for MeIQx-DNA adducts (P < 0.05), liver GST-P(+) cell foci and colon ACF. These results suggest that bLF is a chemopreventive agent for DEN alone or DEN plus MeIQx-induced liver, and MeIQx induced colon carcinogenesis in rats. One possible mechanism is a normalizing down-regulation of CYP1A2 expression by bLF, with consequent reduction of carcinogen activation and adduct formation. PMID- 12079511 TI - Identification of genes associated with dedifferentiation of hepatocellular carcinoma with expression profiling analysis. AB - To identify the genes associated with dedifferentiation of hepatocellular carcinoma (HCC), gene expression profiles of HCCs of well-and moderately differentiated grades were compared by means of oligonucleotide arrays. One tumor showed a nodule-in-nodule appearance (NIN), which is occasionally observed in the course of progression of HCC from well to less differentiated grade, when an inner, moderately differentiated tumor (MD) develops sequentially from the outer, well-differentiated tumor (WD). Seventy-six genes were identified to be up regulated more than 3-fold and 33 genes were down-regulated in the inner nodule in NIN. By statistical analysis of the profiles from 10 individual additional liver tumors, 5 WDs and 5 MDs, we were able to identify 12 genes, LAMA3, PPIB, ADAR, PSMD4, NDUFS8, D9SVA, CCT3, GBAP, ARD1, RDBP, CSRP2, and TLE1, with significantly elevated expression, and 4 genes, CP, IL7R, CD48, and PLGL, with decreased expression in MD. These selected genes were further validated using another 12 tumors, 5 WDs and 7 MDs, with semi-quantitative RT-PCR. We also applied neighborhood analysis to list the genes with high predictability values as most closely correlated with WD-MD distinction. Seven genes, ADAR, PSMD4, D9SVA, CCT3, GBAP, RDBP, and CSRP2, whose expression was elevated and one gene, IL7R, whose expression was decreased, were included among the top 50 predictor genes. These genes are likely to be associated with dedifferentiation of HCC and their identification may help to elucidate the mechanism of liver cancer progression. PMID- 12079510 TI - Inhibitory effects of toremifene on N-methyl-N-nitrosourea and estradiol-17beta induced endometrial carcinogenesis in mice. AB - Short- and long-term experiments were designed to determine the effects of toremifene (TOR) on estrogen-related endometrial carcinogenesis in mice. In the short-term experiment, a single low dose of TOR (0.2 mg / 30 g body weight) decreased expression of c-fos, interleukin (IL)-1alpha, estrogen receptor (ER) alpha mRNAs and corresponding proteins induced by estradiol-17beta (E(2)), in the uteri of the ovariectomized mice. Expression of ER-beta mRNA was increased by the TOR treatment, compared with the control. In the long-term experiment, 106 female ICR mice were given N-methyl-N-nitrosourea (MNU) into their uterine corpora. The animals were divided into four groups as follows: group 1, E(2) diet (5 ppm) plus TOR (0.2 mg / 30 g body weight, subcutaneously, every four weeks); group 2, E(2) diet alone; group 3, basal diet plus TOR. Group 4 served as the control. TOR treatment decreased the incidence of MNU and E(2)-induced endometrial adenocarcinoma and atypical hyperplasia at the termination of the experiment (30 weeks after the start). These results suggest that TOR exerts preventive effects against estrogen-related endometrial carcinogenesis in mice, through the suppression of c-fos as well as IL-1alpha expression induced by E(2). Such suppressive effects of TOR may be related to the decreased ER-alpha and increased ER-beta expressions. PMID- 12079512 TI - Expression of vascular endothelial growth factor and E-cadherin in human ovarian cancer: association with ascites fluid accumulation and peritoneal dissemination in mouse ascites model. AB - Ascites formation and peritoneal dissemination are critical problems in patients with advanced ovarian cancer. Vascular endothelial growth factor (VEGF), also known as angiogenic growth factor, is a potent mediator of peritoneal fluid accumulation and angiogenesis of tumors. E-Cadherin is an adhesion molecule that is important for cell-to-cell interaction. To elucidate the molecular mechanism of ascites formation and peritoneal dissemination of ovarian cancer, we examined the expression of VEGF and E-cadherin in different ovarian cancer cell lines and utilized nude mice to compare the biological characteristics of ovarian cancer cells. Three human ovarian cancer cell lines (AMOC-2, HNOA and HTBOA) were used in this study. Expression of genes was analyzed by northern blotting and RT-PCR methods. AMOC-2 expressed E-cadherin, but not VEGF. HNOA expressed VEGF without E cadherin expression. HTBOA expressed both VEGF and E-cadherin. Each human ovarian cancer model revealed a specific feature. The AMOC-2 mouse had a single large peritoneal tumor without ascites or remarkable peritoneal dissemination. HTBOA and HNOA mice had bloody ascites and marked peritoneal dissemination. Introduction of VEGF antisense into HTBOA cells could inhibit the ascites formation. It is suggested that VEGF is important for the ascites formation via the increased vascular permeability effect. The deregulation of E-cadherin expression might be involved in the peritoneal dissemination. These molecules are important for the formation of specific features of advanced ovarian cancer. Ovarian cancer cell lines that had different gene expression patterns produced nude mouse human ovarian cancer models with different characteristics. PMID- 12079513 TI - Expression of laminin-5 enhances tumorigenicity of human fibrosarcoma cells in nude mice. AB - Laminin-5 (LN5), which consists of laminin alpha3, beta3 and gamma2 chains, is a laminin isoform produced by various kinds of normal epithelial cells and tumor cells. Strong activity of LN5 in adhesion, migration and scattering of cells in vitro and its frequent detection in human tumor tissues have suggested a possible role of LN5 in the malignant growth of tumor cells. To examine whether LN5 affects the malignant potential of tumor cells, we prepared human fibrosarcoma HT1080 cell lines producing LN5 by transfecting a cDNA of laminin alpha3 chain into the parent cell line, which constitutively expressed the laminin beta3 and gamma2 chains. The exogenous alpha3 chain associated with the endogenous beta3 and gamma2 chains to secrete the LN5 heterotrimer that has strong cell-scattering and cell adhesion activities. The HT1080 transfectants expressing LN5 efficiently adhered to culture dishes in a serum-free condition as compared with control HT1080 cells, which secreted the monomers and heterodimer of the beta3 and gamma2 chains. When injected into nude mice subcutaneously, the HT1080 transfectants expressing LN5 grew faster and formed much larger tumors than the control cells. This suggests that LN5 promotes tumor growth in vivo. PMID- 12079514 TI - Expression of PPARgamma and its ligand-dependent growth inhibition in human brain tumor cell lines. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) belongs to a superfamily of thyroid / steroid hormone receptors and regulates transcription of their target genes in a ligand-dependent manner. Recently, PPARgamma was reported to be expressed in several cell lines derived from breast, colon, stomach and lung cancers. Activation of PPARgamma by its ligand inhibits the growth of these tumor cells, suggesting that PPARgamma ligand is a potential anti-cancer agent in PPARgamma-expressing tumors. However, its expression in brain tumors has not been studied. We thus studied the expression in glioma samples with different pathological stages from 20 patients. It was demonstrated that 95% of the glioma tissue expressed PPARgamma mRNA. The results prompted us to study whether PPARgamma ligand affects the growth of cell lines derived from brain tumors. The receptor expression was studied in 9 cell lines either derived from malignant glioma or neuroblastoma. The expression was detected in a glioma cell line SK-MG 1 and in a neuroblastoma cell line NB-1. Addition of one of the PPARgamma ligands, troglitazone, induced growth inhibition in both cell lines. Further analyses revealed that this growth inhibition is caused by a PPARgamma-mediated induction of apoptosis. These results suggest that PPARgamma ligands could be a potential therapeutic agent for the treatment of the brain tumors expressing this receptor. PMID- 12079515 TI - Establishment and characterization of a new lung cancer cell line (MI-4) producing high levels of granulocyte colony stimulating factor. AB - We established a human lung cancer cell line, MI-4 from the pleural effusion of a 69-year-old male with advanced large cell undifferentiated carcinoma of the lung complicated by leukocytosis. The culture supernatant of MI-4 contained high levels of granulocyte colony stimulating factor (G-CSF). The intracellular localization of the G-CSF was identified by immunocytochemistry. Reverse transcription-polymerase chain reaction (RT-PCR) revealed G-CSF mRNA expression in this cell line. The cell line was successfully transplanted into nude mice. The transplanted nude mice also showed leukocytosis with a high serum G-CSF level. Southern blot analysis did not show amplification or rearrangement of the G-CSF gene in MI-4 cells. Spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH) analyses revealed that this cell line has an additional chromosome 17 attached to a segment of chromosome 10 besides two intact chromosomes 17, and that each of these three chromosomes 17 has a G-CSF gene on chromosome 17q. Inflammatory cytokines, tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, significantly enhanced G-CSF expression at both the protein and mRNA levels in MI-4. However, these cytokines did not stimulate the growth of MI-4 cells, regardless of abundant G-CSF production. TNF-alpha rather suppressed it, in a dose-dependent manner. Exogenous recombinant human G-CSF and anti-G-CSF antibody did not promote or inhibit the growth of MI-4 cells at any concentration examined. In addition, RT-PCR analysis did not show G-CSF receptor mRNA expression. These results suggest that this cell line does not have an autocrine growth loop for G-CSF. This cell line should be very useful for understanding the biological activity of G-CSF in G-CSF-overproducing lung cancer. PMID- 12079516 TI - Clinicopathologic comparison between the API2-MALT1 chimeric transcript-positive and -negative gastric low-grade B-cell lymphoma of mucosa-associated lymphoid tissue type. AB - Little is known about the clinicopathological differences between API2-MALT1 chimeric transcript-positive and -negative gastric low-grade B-cell lymphomas of mucosa-associated lymphoid tissue (MALT) type. The aim of this study was to clarify those differences in gastric MALT lymphoma. Twenty-three patients with gastric MALT lymphoma were enrolled in a unicenter study. Helicobacter pylori (H. pylori) infection status and clinical stages were investigated. Antibacterial treatment was performed for every patient. Responsiveness of MALT lymphoma to this treatment was assessed by means of regular follow-up endoscopy combined with biopsy. All cases were examined for API2-MALT1 chimeric transcript by means of RT PCR and sequencing analyses. H. pylori infection status was assessed as positive in 20 patients and negative in three. With regard to responsiveness to antibacterial treatment, complete remission was observed in two patients, partial remission in 12 and no change in nine. API2-MALT1 chimeric transcript was detected in seven patients, all of whom showed no change in response to antibacterial treatment. API2-MALT1 positivity was found to be significantly correlated with responsiveness to antibacterial treatment (P = 0.0001), absence of H. pylori infection (P = 0.0198), and gross cobblestone mucosa observed endoscopically (P = 0.0198). For the other factors (age, sex, dominant site of lesion, high-grade component, infiltrated layer of gastric wall, nodal involvement or clinical stages), there were no differences between API2-MALT1 chimeric transcript-positive and -negative cases. Gastric API2-MALT1 chimeric transcript-positive MALT lymphoma generally features unresponsiveness to antibacterial treatment, and is thought to be unrelated to H. pylori infection in its pathogenesis. Our findings indicate the presence of different clinical subtypes in gastric MALT lymphomas. PMID- 12079518 TI - Clinical significance of occult micrometastases in axillary lymph nodes in "node negative" breast cancer patients. AB - The most important subgroup of breast cancer patients for whom reliable prognostic indicators are needed is women without axillary lymph node metastases. We evaluated the clinical significance of occult micrometastases in axillary lymph nodes in 148 consecutive "node-negative" breast cancer patients. The median age of the patients at surgery was 52 years and the median follow-up period after surgery was 98.5 months. Occult micrometastases were detected in 21 of 148 patients (14.2%) by means of immunohistochemical analysis using AE1 / 3 antibody and a single unstained section after routine histopathological examination. Log rank tests indicated that the 7-year disease-free survival (DFS) and overall survival (OS) rates by Kaplan-Meier methods were significantly better in patients without occult micrometastases than in patients with occult micrometastases [DFS, 93% versus 71% (P = 0.0009); OS, 96% versus 76% (P = 0.0001)]. According to Cox's multivariate analysis, the presence of occult micrometastases had the most significant effect on DFS (P = 0.0053) and OS (P = 0.0035). These findings suggest that the presence of occult micrometastases is an independent and significant predictor of clinical outcome, and that their immunohistochemical detection after routine histopathological examination is useful for selecting the "node-negative" breast cancer patient subgroup at high risk for relapse and death. PMID- 12079519 TI - Immunocytochemical expression of microtubule-associated protein-2 (MAP-2) in small cell lung cancer cell lines with neuronal-like processes. AB - Five of six human small cell lung cancer (SCLC) cell lines changed morphologically into cells with neuronal-like processes on the extracellular matrix of human lung adenocarcinoma cell line PC-9 cells (PC-9 / ECM substrate). The features of the neuronal-like processes of these SCLC cell lines were examined immunocytochemically using monoclonal antibodies against beta-chains of tubulin and microtubule-associated protein-2 (MAP-2), which is somatodendritic MAP of neurons. It was observed that beta-chains of tubulin and MAP-2 were expressed along the neuronal-like processes of SCLC cell lines. These findings suggest that the beta-chains of tubulin and MAP-2 are expressed functionally in SCLC cell lines in association with the development of dendrite-like processes on PC-9 / ECM substrate. PMID- 12079517 TI - MUM1/IRF4 expression is an unfavorable prognostic factor in B-cell chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL). AB - B-Cell chronic lymphocytic leukemia (B-CLL) / small lymphocytic lymphoma (SLL) consists of heterogeneous diseases that are distinguished by morphological, immunophenotypic and molecular features. MUM1 (multiple myeloma oncogene 1) is a protooncogene that is deregulated as a result of (6;14)(p25;q32) chromosomal translocation in multiple myeloma, and is also expressed in a variety of malignant lymphoma entities. We examined the expression of MUM1 in B-CLL / SLL, and found that 2 of 4 B-CLL-derived cell lines and 14 of 29 patients' specimens expressed MUM1 by immunohistochemical analysis. MUM1 expression was not associated with CD38 expression, somatic hypermutation of immunoglobulin heavy chain gene variable region (IgV(H)), or any other clinical characteristics of the patients. Interestingly, the patients who were positive for MUM1 showed shorter overall survival times than those who were negative for MUM1 (50% survival: 22 months vs. 82 months) (P = 0.0008, log-rank test). Multivariate analysis by Cox's proportional-hazards regression model showed that MUM1 expression and unmutated IgV(H) status were independent unfavorable prognostic factors in patients with B CLL / SLL. These findings suggest that MUM1 expression is a useful prognostic factor in B-CLL / SLL. The biological role and mechanism of action of MUM1 in B CLL / SLL need to be clarified for the development of therapies for patients with the poor prognostic subtype. PMID- 12079520 TI - Dose of adenoviral vectors expressing interleukin-2 plays an important role in combined gene therapy with cytosine deaminase/5-fluorocytosine: preclinical consideration. AB - Using a syngeneic murine model, we investigated the therapeutic efficacy of combined gene therapy using adenoviral vectors expressing murine interleukin-2 (AdmIL-2) and Escherichia coli cytosine deaminase (AdCD). In a subcutaneous tumor model, tumor-bearing mice were treated with an intratumoral injection of adenoviral vectors and received an intraperitoneal administration of 5 fluorocytosine (5-FC). Only the mice treated with AdCD (2 x 10(8) pfu) and an intermediate dose of AdmIL-2 (1 x 10(6) pfu) survived significantly longer than mice treated with AdCD alone (P < 0.01). Moreover, 40% of these treated mice obtained complete remission from tumor-bearing status. The cytotoxicity of splenocytes obtained from the treated mice was related to the survival period. Tumor-specific cytotoxic T lymphocyte assay showed that the cell-mediated cytotoxic response was specific for parental tumor cells. In a hepatic metastasis model, mice treated with an intravenous administration of both AdCD (2 x 10(8) pfu) and an intermediate dose of AdmIL-2 (1 x 10(6) pfu) demonstrated the most significant reduction of metastatic foci and the longest survival following a 5 FC administration. These results suggest that gene therapy combined with AdmIL-2 and AdCD may be a promising strategy for clinical application and, in addition, that translation of combined gene therapy from murine models into the clinical setting will require careful attention to the variables of cytokine expression levels in the design of clinical trials and in the evaluation of treatment efficacy. PMID- 12079521 TI - Hammerhead ribozyme against gamma-glutamylcysteine synthetase attenuates resistance to ionizing radiation and cisplatin in human T98G glioblastoma cells. AB - Glioblastoma cells are highly malignant and show resistance to ionizing radiation, as well as anti-cancer drugs. This resistance to cancer therapy is often associated with a high concentration of glutathione (GSH). In this study, the effect of continuous down-regulation of gamma-glutamylcysteine synthetase (gamma-GCS) expression, a rate-limiting enzyme for GSH synthesis, on resistance to ionizing radiation and cisplatin (CDDP) was studied in T98G human glioblastoma cells. We constructed a hammerhead ribozyme against a gamma-GCS heavy subunit (gamma-GCSh) mRNA and transfected it into T98G cells. (1) The transfection of the ribozyme decreased the concentration of GSH and resulted in G1 cell cycle arrest of T98G cells. (2) The transfection of the ribozyme increased the cytotoxicity of ionizing radiation and CDDP in T98G cells. Thus, hammerhead ribozyme against gamma-GCS is suggested to have potential as a cancer gene therapy to reduce the resistance of malignant cells to ionizing radiation and anti-cancer drugs. PMID- 12079522 TI - Sensitivity to anticancer agents and resistance mechanisms in clear cell carcinoma of the ovary. AB - We conducted the present study to determine the chemoresistance mechanisms in clear cell carcinoma of the ovary (CCC). Five human CCC cell lines (HAC-2, RMG-I, RMG-II, KK, and KOC-7c) were used in this study. The sensitivity of the cells to the anticancer agents was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay and we assessed drug sensitivity by calculating assay area under the curve (AUC) for each agent. The expression of multi-drug resistance genes (MDR-1, MRP-1, MRP-2) was detected by reverse transcription-polymerase chain reaction (RT-PCR). Glutathione (GSH) concentration was measured by an enzymatic assay. Topoisomerase (topo) I activity was assayed in terms of relaxation of supercoiled plasmid substrate DNA. The IC(50) to anticancer agents ranged widely. The assay AUC indicated that 3 of 5 cell lines (RMG-I, RMG-II, and KK) were sensitive to paclitaxel (PTX), 3 (HAC-2, RMG-I, and RMG-II) were sensitive to 7-ethyl-10-hydroxycamptothecin (SN-38), which is an active metabolite of camptothecin (CPT-11), and only one (HAC-2) was sensitive to cisplatin (CDDP). All cell lines were resistant to mitomycin-C (MMC) and etoposide (VP-16). The MRP-1 gene was detected in all cell lines. Only one cell line showed both MRP-2 and MDR-1 gene expression. Except for HAC-2 cells, expression of MRP genes was related to CDDP resistance, and MDR-1 gene expression was associated with PTX resistance. GSH concentrations increased after exposure to CDDP or MMC in all cell lines. There was a significant correlation between topo-I enzymatic activity and the response to SN-38. The present study revealed several resistance mechanisms in CCC and the results suggested that PTX and CPT 11 might be effective agents to treat CCC. PMID- 12079523 TI - The fractal nature of nature: power laws, ecological complexity and biodiversity. AB - Underlying the diversity of life and the complexity of ecology is order that reflects the operation of fundamental physical and biological processes. Power laws describe empirical scaling relationships that are emergent quantitative features of biodiversity. These features are patterns of structure or dynamics that are self-similar or fractal-like over many orders of magnitude. Power laws allow extrapolation and prediction over a wide range of scales. Some appear to be universal, occurring in virtually all taxa of organisms and types of environments. They offer clues to underlying mechanisms that powerfully constrain biodiversity. We describe recent progress and future prospects for understanding the mechanisms that generate these power laws, and for explaining the diversity of species and complexity of ecosystems in terms of fundamental principles of physical and biological science. PMID- 12079524 TI - Scaling in the growth of geographically subdivided populations: invariant patterns from a continent-wide biological survey. AB - We consider statistical patterns of variation in growth rates for over 400 species of breeding birds across North America surveyed from 1966 to 1998. We report two results. First, the standard deviation of population growth rates decays as a power-law function of total population size with an exponent beta = 0.36 +/- 0.02. Second, the number of subpopulations, measured as the number of survey locations with non-zero counts, scales to the 3/4 power of total number of birds counted in a given species. We show how these patterns may be related, and discuss a simple stochastic growth model for a geographically subdivided population that formalizes the relationship. We also examine reasons that may explain why some species deviate from these scaling laws. PMID- 12079526 TI - Dynamics of climate and ecosystem coupling: abrupt changes and multiple equilibria. AB - Interactions between subunits of the global climate-biosphere system (e.g. atmosphere, ocean, biosphere and cryosphere) often lead to behaviour that is not evident when each subunit is viewed in isolation. This newly evident behaviour is an emergent property of the coupled subsystems. Interactions between thermohaline circulation and climate illustrate one emergent property of coupling ocean and atmospheric circulation. The multiple thermohaline circulation equilibria that result caused abrupt climate changes in the past and may cause abrupt climate changes in the future. Similarly, coupling between the climate system and ecosystem structure and function produces complex behaviour in certain regions. For example, atmosphere-biosphere interactions in the Sahel region of West Africa lead to multiple stable equilibria. Either wet or dry climate equilibria can occur under otherwise identical forcing conditions. The equilibrium reached is dependent on past history (i.e. initial conditions), and relatively small perturbations to either climate or vegetation can cause switching between the two equilibria. Both thermohaline circulation and the climate-vegetation system in the Sahel are prone to abrupt changes that may be irreversible. This complicates the relatively linear view of global changes held in many scientific and policy communities. Emergent properties of coupled socio-natural systems add yet another layer of complexity to the policy debate. As a result, the social and economic consequences of possible global changes are likely to be underestimated in most conventional analyses because these nonlinear, abrupt and irreversible responses are insufficiently considered. PMID- 12079525 TI - Multiple states in river and lake ecosystems. AB - Nonlinear models of ecosystem dynamics that incorporate positive feedbacks and multiple, internally reinforced states have considerable explanatory power. However, linear models may be adequate, particularly if ecosystem behaviour is primarily controlled by external processes. In lake ecosystems, internal (mainly biotic) processes are thought to have major impacts on system behaviour, whereas in rivers, external (mainly physical) factors have traditionally been emphasized. We consider the hypothesis that models that exhibit multiple states are useful for understanding the behaviour of lake ecosystems, but not as useful for understanding stream ecosystems. Some of the best-known examples of multiple states come from lake ecosystems. We review some of these examples, and we also describe examples of multiple states in rivers. We conclude that the hypothesis is an oversimplification; the importance of physical forcing in rivers does not eliminate the possibility of internal feedbacks that create multiple states, although in rivers these feedbacks are likely to include physical as well as biotic processes. Nonlinear behaviour in aquatic ecosystems may be more common than current theory indicates. PMID- 12079527 TI - Cluster size distributions: signatures of self-organization in spatial ecologies. AB - Three different lattice-based models for antagonistic ecological interactions, both nonlinear and stochastic, exhibit similar power-law scalings in the geometry of clusters. Specifically, cluster size distributions and perimeter-area curves follow power-law scalings. In the coexistence regime, these patterns are robust: their exponents, and therefore the associated Korcak exponent characterizing patchiness, depend only weakly on the parameters of the systems. These distributions, in particular the values of their exponents, are close to those reported in the literature for systems associated with self-organized criticality (SOC) such as forest-fire models; however, the typical assumptions of SOC need not apply. Our results demonstrate that power-law scalings in cluster size distributions are not restricted to systems for antagonistic interactions in which a clear separation of time-scales holds. The patterns are characteristic of processes of growth and inhibition in space, such as those in predator-prey and disturbance-recovery dynamics. Inversions of these patterns, that is, scalings with a positive slope as described for plankton distributions, would therefore require spatial forcing by environmental variability. PMID- 12079528 TI - Self-organized instability in complex ecosystems. AB - Why are some ecosystems so rich, yet contain so many rare species? High species diversity, together with rarity, is a general trend in neotropical forests and coral reefs. However, the origin of such diversity and the consequences of food web complexity in both species abundances and temporal fluctuations are not well understood. Several regularities are observed in complex, multispecies ecosystems that suggest that these ecologies might be organized close to points of instability. We explore, in greater depth, a recent stochastic model of population dynamics that is shown to reproduce: (i) the scaling law linking species number and connectivity; (ii) the observed distributions of species abundance reported from field studies (showing long tails and thus a predominance of rare species); (iii) the complex fluctuations displayed by natural communities (including chaotic dynamics); and (iv) the species-area relations displayed by rainforest plots. It is conjectured that the conflict between the natural tendency towards higher diversity due to immigration, and the ecosystem level constraints derived from an increasing number of links, leaves the system poised at a critical boundary separating stable from unstable communities, where large fluctuations are expected to occur. We suggest that the patterns displayed by species-rich communities, including rarity, would result from such a spontaneous tendency towards instability. PMID- 12079529 TI - Gaia as a complex adaptive system. AB - We define the Gaia system of life and its environment on Earth, review the status of the Gaia theory, introduce potentially relevant concepts from complexity theory, then try to apply them to Gaia. We consider whether Gaia is a complex adaptive system (CAS) in terms of its behaviour and suggest that the system is self-organizing but does not reside in a critical state. Gaia has supported abundant life for most of the last 3.8 Gyr. Large perturbations have occasionally suppressed life but the system has always recovered without losing the capacity for large-scale free energy capture and recycling of essential elements. To illustrate how complexity theory can help us understand the emergence of planetary-scale order, we present a simple cellular automata (CA) model of the imaginary planet Daisyworld. This exhibits emergent self-regulation as a consequence of feedback coupling between life and its environment. Local spatial interaction, which was absent from the original model, can destabilize the system by generating bifurcation regimes. Variation and natural selection tend to remove this instability. With mutation in the model system, it exhibits self-organizing adaptive behaviour in its response to forcing. We close by suggesting how artificial life ('Alife') techniques may enable more comprehensive feasibility tests of Gaia. PMID- 12079530 TI - Recovery after mass extinction: evolutionary assembly in large-scale biosphere dynamics. AB - Biotic recoveries following mass extinctions are characterized by a process in which whole ecologies are reconstructed from low-diversity systems, often characterized by opportunistic groups. The recovery process provides an unexpected window to ecosystem dynamics. In many aspects, recovery is very similar to ecological succession, but important differences are also apparently linked to the innovative patterns of niche construction observed in the fossil record. In this paper, we analyse the similarities and differences between ecological succession and evolutionary recovery to provide a preliminary ecological theory of recoveries. A simple evolutionary model with three trophic levels is presented, and its properties (closely resembling those observed in the fossil record) are compared with characteristic patterns of ecological response to disturbances in continuous models of three-level ecosystems. PMID- 12079531 TI - Does natural selection organize ecosystems for the maintenance of high productivity and diversity? AB - Three types of evidence suggest that natural ecosystems are organized for high productivity and diversity: (i) changes not previously experienced by a natural ecosystem, such as novel human disturbances, tend to diminish its productivity and/or diversity, just as 'random' changes in a machine designed for a function usually impair its execution of that function; (ii) humans strive to recreate properties of natural ecosystems to enhance productivity of artificial ones, as farmers try to recreate properties of natural soils in their fields; and (iii) productivity and diversity have increased during the Earth's history as a whole, and after every major biotic crisis. Natural selection results in ecosystems organized to maintain high productivity of organic matter and diversity of species, just as competition among individuals in Adam Smith's ideal economy favours high production of wealth and diversity of occupations. In nature, poorly exploited energy attracts more efficient users. This circumstance favours the opening of new ways of life and more efficient recycling of resources, and eliminates most productivity-reducing 'ecological monopolies'. Ecological dominants tend to be replaced by successors with higher metabolism, which respond to more stimuli and engage in more varied interactions. Finally, increasingly efficient predators and herbivores favour faster turnover of resources. PMID- 12079532 TI - Rangelands, pastoralists and governments: interlinked systems of people and nature. AB - We analyse commercially operated rangelands as coupled systems of people and nature. The biophysical components include: (i) the reduction and recovery of potential primary production, reflected as changes in grass production per unit of rainfall; (ii) changes in woody plants dependent on the grazing and fire regimes; and (iii) livestock and wool dynamics influenced by season, condition of the rangeland and numbers of wild and feral animals. The social components include the managers, who vary with regard to a range of cognitive abilities and lifestyle choices, and the regulators who vary in regard to policy goals. We compare agent-based and optimization models of a rangeland system. The agent based model leads to recognition that policies select for certain management practices by creating a template that governs the trajectories of the behaviour of individuals, learning, and overall system dynamics. Conservative regulations reduce short-term loss in production but also restrict learning. A free-market environment leads to severe degradation but the surviving pastoralists perform well under subsequent variable conditions. The challenge for policy makers is to balance the needs for learning and for preventing excessive degradation. A genetic algorithm model optimizing for net discounted income and based on a population of management solutions (stocking rate, how much to suppress fire, etc.) indicates that robust solutions lead to a loss of about 40% compared with solutions where the sequence of rainfall was known in advance: this is a similar figure to that obtained from the agent-based model. We conclude that, on the basis of Levin's three criteria, rangelands with their livestock and human managers do constitute complex adaptive systems. If this is so, then command-and control approaches to rangeland policy and management are bound to fail. PMID- 12079533 TI - Modelling radiation-induced chromosome aberrations. AB - PURPOSE: To review the data and models on the induction of chromosomal aberrations by a single acute dose of radiation, then critically to assess progress that has been made so far to make a judgement about prospects for increased understanding. MATERIALS AND METHODS: Drawing upon many years of personal experience, a consistent set of data on the radiation-induced dicentric yield in human lymphocytes was produced. Other data concerning the production of complex rearrangements and the time-course of the appearance of dicentrics has also been reviewed. Three basic models have been reviewed and compared with respect to their ability to predict the judged dose-effect relationships as well as the time-course and complexity of the aberration production. CONCLUSIONS: None of the three basic models alone can explain adequately all of the available data. A combination of the models has succeeded to a greater extent, but there is still no complete agreement. The major reason for the discrepancy between the measured yield curves and their predictions lies in the variation of the quadratic term in the dose-response relationship with increasing linear energy transfer. Further experimental work is required to resolve this. PMID- 12079534 TI - Role of damage-sensing/processing genes in the radiation response of haemopoietic in vitro colony-forming cells. AB - PURPOSE: To characterize the role of various cellular damagesensing, processing and survival genes in the in-vitro radiosensitivity of haemopoietic colony forming cells. MATERIALS AND METHODS: Bone marrow cells from a range of different gene-knockout mice were irradiated in vitro with graded radiation doses and assayed for colony-forming efficiency. RESULTS: Colony-forming efficiency in the nulls was often lower by up to threefold compared with the wild-types. This was noticeable in particular for the atm, bax and p21 nulls. Radiosensitivity was markedly increased in the scid mouse (about 2.3-fold), more than in the atm null mouse (about 1.7-fold). There was resistance in the p53 nulls compared with the wild-types, using two different background strains, that gave similar results. There was slight sensitization in the p21 nulls. In the bcl-2 nulls, there was sensitization at low dose, but not at high dose. In contrast, in the bax nulls, there was protection at low dose, but again not at high dose. The heterozygotes for p53, bcl-2 and bax responded similarly to the wild types, so that no gene dosage effects were identified. CONCLUSIONS: These studies are the first to elucidate the role of as many as six relevant genes in the radiosensitivity of a single cell type. They show the greater importance of 'survival' genes at lower cytotoxic doses of radiation compared with the greater importance of 'damage sensing' genes at higher doses. PMID- 12079535 TI - The effects of low-dose X-irradiation on the oxidative burst in stimulated macrophages. AB - PURPOSE: Local irradiation with a dose of around 0.5 Gy is an effective treatment of acute necrotizing inflammations. The hypothesis that low doses of X-rays modulate the oxidative burst in activated macrophages, which plays a major role in the acute inflammatory process, was tested. MATERIALS AND METHODS: Murine RAW 264.7 macrophages were stimulated with LPS/gammaIFN, PMA or zymosan and oxidative burst was measured using either DCFH-DA or by reduction of cytochrome-C. Radiation doses of 0.3-10 Gy were given shortly before or after stimulation. RESULTS: Low X-ray doses of <1 Gy significantly reduced the oxidative burst in activated macrophages, whereas higher doses had little effect on oxidative burst. CONCLUSIONS: The modulation of oxidative burst by low radiation doses may contribute to the therapeutic effectiveness of low-dose radiotherapy of acute necrotizing inflammations. PMID- 12079536 TI - Protein kinase inhibitors modulate time-dependent effects of UV and ionizing irradiation on ICAM-1 expression on human hepatoma cells. AB - PURPOSE: To investigate modulation of the expression of the adhesion protein ICAM 1 by UV and ionizing irradiation. MATERIALS AND METHODS: HepG2 hepatoma cells were irradiated in vitro with UVB (20 mJ cm(-2)) or X-rays (5 Gy), respectively. Gene expression of ICAM-1 after irradiation was quantified by RT-PCR. Cell surface density of ICAM-1 was determined by flow cytometry. Protein or lipid kinase inhibitors were used to clarify radiation-induced transduction pathways that control ICAM-1 expression. Immuno-electron microscopy and dot-blot analysis were used to examine localization of ICAM-1. RESULTS: The study showed time dependent effects of ionizing and UV irradiation on ICAM-1 expression of HepG2 cells. After an immediate transient decrease of ICAM-1 cell surface expression within minutes to hours, ICAM-1 expression increased up to 1.35-fold over normal level at 48 h post-irradiation. Irradiation caused ICAM-1 to become internalized into lysosomes. Additionally, ICAM-1 together with parts of the cell were pinched off. Finally, ICAM-1 levels were down- and up-regulated by decreased or increased gene expression. The early decrease of ICAM-1 expression could be blocked by a potent PKC inhibitor (BisX), whereas the increase of ICAM-1 after 24 h was prevented by addition of the p38 MAP kinase inhibitor SB 203580. CONCLUSION: The data suggest that ICAM-1 expression is modulated by UV, as well as ionizing radiation, in a time-dependent way involving PKC and p38 MAP kinase pathways. PMID- 12079537 TI - Characterization of 2-hydroxyadenine DNA glycosylase activity of Escherichia coli MutY protein. AB - PURPOSE: 2-Hydroxyadenine (2-ohA) is an oxidation product of adenine generated in DNA by ionizing radiation and various chemical oxidants. 2-ohA has mutational potential comparable to that of 8-oxoguanine in bacteria and mammalian cells. Recent studies have shown that 2-ohA is removed from DNA by a human MutY homolog, MYH protein, in vitro. On the other hand, the repair mechanisms for 2-ohA in Escherichia coli are not yet understood. MATERIALS AND METHODS: Gel shift assays were used to assess the binding activity of E. coli full-length MutY protein and its N-terminal (residues 1-226) domain (M25) to 2-ohA/G-, 2-ohA/A-, 2-ohA/C- and 2-ohA/T-containing 24-mer oligonucleotides. Furthermore, whether these proteins specifically cleave 2-ohA-containing duplex oligonucleotides was examined. RESULTS: The purified MutY and M25 proteins had similar binding affinities to 2 ohA/G-, 2-ohA/A- and 2-ohA/C-containing oligonucleotides. MutY protein removed 2 ohA preferentially from 2-ohA/G mispairs. M25 protein showed the reduced catalytic activity for 2-ohA/G-containing oligonucleotides. CONCLUSIONS: E. coli MutY protein has a DNA glycosylase activity that removes 2-ohA from 2-ohA/G mispairs in DNA. The C-terminal domain is required for the removal of 2-ohA from DNA, but is not crucial for binding to 2-ohA-containing oligonucleotides. PMID- 12079538 TI - Do low dose-rate bystander effects influence domestic radon risks? AB - PURPOSE: Radon risks derive from exposure of bronchio-epithelial cells to high linear energy transfer (LET) alpha-particles. alpha-particle exposure can result in bystander effects, where irradiated cells emit signals resulting in damage to nearby unirradiated bystander cells. This can result in non-linear dose-response relations, and inverse dose-rate effects. Domestic radon risk estimates are currently extrapolated from miner data, which are at both higher doses and higher dose-rates, so bystander effects on unhit cells could play a large role in the extrapolation of risks from mines to homes. Therefore, we extend an earlier quantitative mechanistic model of bystander effects to include protracted exposure, with the aim of quantifying the significance of the bystander effect for very prolonged exposures. MATERIALS AND METHODS: A model of high-LET bystander effects, originally developed to analyse oncogenic transformation in vitro, is extended to low dose-rates. The model considers radiation response as a superposition of bystander and linear direct e It attributes bystander effects to a small subpopulation of hypersensitive cells, with the bystander contribution dominating the direct contribution at very low acute doses but saturating as the dose increases. Inverse dose-rate effects are attributed to the replenishment of the hypersensitive subpopulation during prolonged irradiation. RESULTS: The model was fitted to dose- and dose-rate-dependent radon-exposed miner data, suggesting that one directly hit target bronchio-epithelial cell can send bystander signals to about 50 neighbouring target cells. The model suggests that a naive linear extrapolation of radon miner data to low doses, without accounting for dose-rate, would result in an underestimation of domestic radon risks by about a factor of 4, a value comparable with the empirical estimate applied in the recent BEIR-VI report on radon risk estimation. CONCLUSIONS: Bystander effects represent a plausible quantitative and mechanistic explanation of inverse dose-rate effects by high-LET radiation, resulting in non-linear dose-response relations and a complex interplay between the effects of dose and exposure time. The model presented provides a potential mechanistic underpinning for the empirical exposure-time correction factors applied in the recent BEIR-VI for domestic radon risk estimation. PMID- 12079539 TI - Alpha-particle lineal energy spectra for the human lung. AB - PURPOSE: To determine the distribution of lineal energy in the target cells and the parameters of distribution for various combinations of sources, targets and energies in the human tracheobronchial tree. Frequency average and average of the square of the lineal energy were also calculated from the lineal energy distribution. METHOD: A model was created to simulate the geometric distribution of cell nuclei in the airway of the tracheobronchial tree. Propagation of alpha particles in such a model was simulated by the Monte Carlo method. Microdosimetric distributions of lineal energy were calculated. RESULTS: Distributions of lineal energy were substantially different for basal and secretory cells. The frequency average of specific energy was found to be between 0.3 Gy and 0.4 Gy. CONCLUSIONS: Interactions of alpha particles with basal and secretory cells show significant differences: events with small energy transfer are dominant for basal cells, while those with large transfer are dominant for secretory cells. This finding can influence dose weightings and yield lower values of the dose-to-exposure coefficient. PMID- 12079540 TI - In vitro radiosensitivity measured in lymphocytes and fibroblasts by colony formation and comet assay: comparison with clinical acute reactions to radiotherapy in breast cancer patients. AB - PURPOSE: To compare colony-forming and comet assays on fibroblasts and lymphocytes of 32 breast cancer patients irradiated after breast-conserving operations and to correlate the results with acute clinical radiation reactions in the skin. MATERIAL AND METHODS: Skin fibroblasts were isolated and cultivated before radiotherapy and lymphocytes were drawn prior to the first and directly after the final external irradiation. The colony-forming assay was performed with fibroblasts and the comet assay with lymphocytes and fibroblasts of breast cancer patients according to standard protocols. The clinical radiation reactions of the patients were graded according to the RTOG system. RESULTS: No significant correlation (p =0.09) was detected between clinical acute skin reactions and the in vitro clonogenic data in fibroblasts. Results of the comet assay in lymphocytes, however, showed a significant correlation (p <0.05) with the clinical data when patients were divided into two groups with average and elevated acute reactions. Apart from initial damage, fibroblasts did not show significant differences between the two patient groups. Repeated comet assays in lymphocytes of the same patient drawn before treatment and before and after external radiotherapy demonstrated good reproducibility of the test and no significant impact of preceding radiation treatment. There was a good correlation (r =0.65) between the comet assay results in fibroblasts and lymphocytes of the same individual. CONCLUSIONS: In this cohort of patients, a significant correlation between the in vitro results of the comet assay in lymphocytes and clinical acute reactions was detected. The results of the comet assay and of fibroblast colony formation did not correlate with in vitro radiosensitivity. PMID- 12079541 TI - Dose-dependent differential effects of low and pulsed dose-rate brachytherapy in a radioresistant syngenic rat prostate tumour model. AB - PURPOSE: To study the response of the Dunning prostate carcinoma (R3327-AT1 subline) to continuous low dose-rate (CLDR) and pulsed dose-rate (PDR) brachytherapy. MATERIALS AND METHODS: After subcutaneous tumour transplantation into the thigh of the Copenhagen rat, doses of 0, 20, 30, 40 and 50 Gy were applied to the tumour surface (tumour diameter 9+/-1mm). Eight animals were irradiated per dose group and exposure condition. Interstitial PDR ((192)Ir source, 37 GBq) and CLDR ((192)Ir seed, 150 MBq) brachytherapy were carried out with 0.75 Gy/pulse h(-1) and a dose-rate of 0.75Gyh(-1), respectively. Treatment response was assessed in terms of growth delay expressed as the time (T(5)) required for each tumour to reach five times the initial tumour volume. RESULTS: The median T(5) times for the CLDR groups (in the order: control, 20, 30, 40, 50 Gy) were 12 (12), 54.5 (21), 64.5 (31), 85.5 (51), and 65 (47.5) days. Values after PDR brachytherapy are given in parentheses and resulted in a significantly impaired tumour growth delay (log-rank test) in the 20Gy (p =0.006) and 30 Gy (p =0.036) groups. No significant difference was found in the 40-50 Gy dose range. CONCLUSIONS: In contrast to previous results and predictions of biological models we observed dose-dependent differential effects of PDR and CLDR brachytherapy with reduced efficacy of PDR in the lower dose range. PMID- 12079542 TI - Evaluation of the frequencies of chromosomal aberrations in a population exposed to prolonged low dose-rate 60Co gamma-irradiation. AB - PURPOSE: Chromosomal aberration analysis in peripheral blood lymphocytes was performed to evaluate late cytogenetic effects of long-term low dose-rate gamma irradiation exposure among students and residents exposed in radiocontaminated buildings. MATERIALS AND METHODS: Blood samples were taken from 1913 subjects (age 17.8+/-13.6, mean+/-SD) 5-8 years after their relocation from radioactive environments as well as from 176 non-exposed subjects (age 29.6+/-11.9) from the local community. Their lymphocytes were cultured for 48 h and metaphase spreads were prepared. A total of 208 900 metaphases were analysed for different types of chromosomal aberrations. RESULTS: Relatively higher frequencies of translocations (2.1 x 10(-3)), rings (0.6 x 10(-3)) and dicentrics (0.6 x 10(-3)) were noted in the exposed population as compared with the nonexposed reference populations. Moreover, 356 (78.6%) of the 453 inversions were found on 14q11.2q32 in the exposed population. Among 392 well-demonstrated translocations, 167 (42.6%) and 175 (44.6%) occurred in chromosomes 7 and 14, respectively, while 139 (35.5%) occurred as t(7;14). In particular, the aberrations t(7;14)(p13;q11.2), t(7;14)(p15;q11.2) and t(7;14)(q36;q11.2) were the most prevalent, occurring with frequencies of 19 (13.7%), 20 (14.4%) and 27 (19.4%), respectively. In these, 3205 breakpoints were documented, with chromosomes 7, 9 and 14 shown to carry significantly higher frequencies of breakpoints than expected (chi(2)-test, p<0.0001). A further six hotspots were identified on 7p15 (57, 1.8%), 7q36 (42, 1.3%), 9q12 (244, 7.6%), 9q13 (86, 2.7%), 14q11.2 (509, 15.9%) and 14q32 (387, 12.1%) in the exposed population. CONCLUSION: In comparison with the unexposed population, we observed increased frequencies of various chromosomal aberrations in this human population with previous exposure to prolonged low dose-rate gamma radiation. Moreover, several hotspot breakpoints and inversions and translocations were observed on chromosomes 7 and 14. PMID- 12079543 TI - Health examination and chromosome aberration analysis of residents living in 60Co contaminated rebar buildings. AB - PURPOSE: To elucidate the results of health examination and chromosome aberration analysis of residents living in (60)Co-contaminated rebar buildings. MATERIAL AND METHODS: Medical surveillance including peripheral blood analysis, urine analysis, chest X-ray, blood biochemical analysis, electrocardiography, thyroid function test and physical examination was performed on 189 residents within 6 months after disclosure of radiation exposure. The annual dose for this group of residents varied from 2 to 95 mSv year(-1) (mean+/-1 SD = 18+/-21 mSv year(-1)) above the background exposure of the Taiwanese population. For chromosome analysis, 500-671 lymphocytes were scored for 136 residents, and the numbers of dicentrics and rings were counted and analysed. RESULTS: Medical surveillance showed that the most frequent abnormalities were elevation of the erythrocyte sedimentation rate occurring in 14.8% of those examined. Reduction of the negative charge on erthrocytes by ionizing radiation may cause an elevation of erythrocyte sedimentation rate. Statistical analysis of chromosome aberration data showed that the mean dicentric frequency for radiation-contaminated building residents (0.69+/-0.93 SD) was significantly higher than those for controls (0.33+/-0.49) (p <0.01). CONCLUSIONS: No radiation effects were found in the preliminary health examination. However, lymphocyte chromosome analysis of the radiation-contaminated building residents showed that prolonged low-dose-rate gamma exposure induced cytogenetic changes in humans. PMID- 12079544 TI - A Monte Carlo EM approach for partially observable diffusion processes: theory and applications to neural networks. AB - We present a Monte Carlo approach for training partially observable diffusion processes. We apply the approach to diffusion networks, a stochastic version of continuous recurrent neural networks. The approach is aimed at learning probability distributions of continuous paths, not just expected values. Interestingly, the relevant activation statistics used by the learning rule presented here are inner products in the Hilbert space of square integrable functions. These inner products can be computed using Hebbian operations and do not require backpropagation of error signals. Moreover, standard kernel methods could potentially be applied to compute such inner products. We propose that the main reason that recurrent neural networks have not worked well in engineering applications (e.g., speech recognition) is that they implicitly rely on a very simplistic likelihood model. The diffusion network approach proposed here is much richer and may open new avenues for applications of recurrent neural networks. We present some analysis and simulations to support this view. Very encouraging results were obtained on a visual speech recognition task in which neural networks outperformed hidden Markov models. PMID- 12079545 TI - Are visual cortex maps optimized for coverage? AB - The elegant regularity of maps of variables such as ocular dominance, orientation, and spatial frequency in primary visual cortex has prompted many people to suggest that their structure could be explained by an optimization principle. Up to now, the standard way to test this hypothesis has been to generate artificial maps by optimizing a hypothesized objective function and then to compare these artificial maps with real maps using a variety of quantitative criteria. If the artificial maps are similar to the real maps, this provides some evidence that the real cortex may be optimizing a similar function to the one hypothesized. Recently, a more direct method has been proposed for testing whether real maps represent local optima of an objective function (Swindale, Shoham, Grinvald, Bonhoeffer, & Hubener, 2000). In this approach, the value of the hypothesized function is calculated for a real map, and then the real map is perturbed in certain ways and the function recalculated. If each of these perturbations leads to a worsening of the function, it is tempting to conclude that the real map is quite likely to represent a local optimum of that function. In this article, we argue that such perturbation results provide only weak evidence in favor of the optimization hypothesis. PMID- 12079546 TI - Kernel-based topographic map formation by local density modeling. AB - We introduce a new learning algorithm for kernel-based topographic map formation. The algorithm generates a gaussian mixture density model by individually adapting the gaussian kernels' centers and radii to the assumed gaussian local input densities. PMID- 12079547 TI - A simple model of long-term spike train regularization. AB - A simple model of spike generation is described that gives rise to negative correlations in the interspike interval (ISI) sequence and leads to long-term spike train regularization. This regularization can be seen by examining the variance of the kth-order interval distribution for large k (the times between spike i and spike i + k). The variance is much smaller than would be expected if successive ISIs were uncorrelated. Such regularizing effects have been observed in the spike trains of electrosensory afferent nerve fibers and can lead to dramatic improvement in the detectability of weak signals encoded in the spike train data (Ratnam & Nelson, 2000). Here, we present a simple neural model in which negative ISI correlations and long-term spike train regularization arise from refractory effects associated with a dynamic spike threshold. Our model is derived from a more detailed model of electrosensory afferent dynamics developed recently by other investigators (Chacron, Longtin, St.-Hilaire, & Maler, 2000;Chacron, Longtin, & Maler, 2001). The core of this model is a dynamic spike threshold that is transiently elevated following a spike and subsequently decays until the next spike is generated. Here, we present a simplified version-the linear adaptive threshold model-that contains a single state variable and three free parameters that control the mean and coefficient of variation of the spontaneous ISI distribution and the frequency characteristics of the driven response. We show that refractory effects associated with the dynamic threshold lead to regularization of the spike train on long timescales. Furthermore, we show that this regularization enhances the detectability of weak signals encoded by the linear adaptive threshold model. Although inspired by properties of electrosensory afferent nerve fibers, such regularizing effects may play an important role in other neural systems where weak signals must be reliably detected in noisy spike trains. When modeling a neuronal system that exhibits this type of ISI correlation structure, the linear adaptive threshold model may provide a more appropriate starting point than conventional renewal process models that lack long-term regularizing effects. PMID- 12079548 TI - Spatiotemporal spike encoding of a continuous external signal. AB - Interspike intervals of spikes emitted from an integrator neuron model of sensory neurons can encode input information represented as a continuous signal from a deterministic system. If a real brain uses spike timing as a means of information processing, other neurons receiving spatiotemporal spikes from such sensory neurons must also be capable of treating information included in deterministic interspike intervals. In this article, we examine functions of neurons modeling cortical neurons receiving spatiotemporal spikes from many sensory neurons. We show that such neuron models can encode stimulus information passed from the sensory model neurons in the form of interspike intervals. Each sensory neuron connected to the cortical neuron contributes equally to the information collection by the cortical neuron. Although the incident spike train to the cortical neuron is a superimposition of spike trains from many sensory neurons, it need not be decomposed into spike trains according to the input neurons. These results are also preserved for generalizations of sensory neurons such as a small amount of leak, noise, inhomogeneity in firing rates, or biases introduced in the phase distributions. PMID- 12079549 TI - Attractor reliability reveals deterministic structure in neuronal spike trains. AB - When periodic current is injected into an integrate-and-fire model neuron, the voltage as a function of time converges from different initial conditions to an attractor that produces reproducible sequences of spikes. The attractor reliability is a measure of the stability of spike trains against intrinsic noise and is quantified here as the inverse of the number of distinct spike trains obtained in response to repeated presentations of the same stimulus. High reliability characterizes neurons that can support a spike-time code, unlike neurons with discharges forming a renewal process (such as a Poisson process). These two classes of responses cannot be distinguished using measures based on the spike-time histogram, but they can be identified by the attractor dynamics of spike trains, as shown here using a new method for calculating the attractor reliability. We applied these methods to spike trains obtained from current injection into cortical neurons recorded in vitro. These spike trains did not form a renewal process and had a higher reliability compared to renewal-like processes with the same spike-time histogram. PMID- 12079550 TI - Traveling waves of excitation in neural field models: equivalence of rate descriptions and integrate-and-fire dynamics. AB - Field models provide an elegant mathematical framework to analyze large-scale patterns of neural activity. On the microscopic level, these models are usually based on either a firing-rate picture or integrate-and-fire dynamics. This article shows that in spite of the large conceptual differences between the two types of dynamics, both generate closely related plane-wave solutions. Furthermore, for a large group of models, estimates about the network connectivity derived from the speed of these plane waves only marginally depend on the assumed class of microscopic dynamics. We derive quantitative results about this phenomenon and discuss consequences for the interpretation of experimental data. PMID- 12079551 TI - Attentional recruitment of inter-areal recurrent networks for selective gain control. AB - There is strong anatomical and physiological evidence that neurons with large receptive fields located in higher visual areas are recurrently connected to neurons with smaller receptive fields in lower areas. We have previously described a minimal neuronal network architecture in which top-down attentional signals to large receptive field neurons can bias and selectively read out the bottom-up sensory information to small receptive field neurons (Hahnloser, Douglas, Mahowald, & Hepp, 1999). Here we study an enhanced model, where the role of attention is to recruit specific inter-areal feedback loops (e.g., drive neurons above firing threshold). We first illustrate the operation of recruitment on a simple example of visual stimulus selection. In the subsequent analysis, we find that attentional recruitment operates by dynamical modulation of signal amplification and response multistability. In particular, we find that attentional stimulus selection necessitates increased recruitment when the stimulus to be selected is of small contrast and of small distance away from distractor stimuli. The selectability of a low-contrast stimulus is dependent on the gain of attentional effects; for example, low-contrast stimuli can be selected only when attention enhances neural responses. However, the dependence of attentional selection on stimulus-distractor distance is not contingent on whether attention enhances or suppresses responses. The computational implications of attentional recruitment are that cortical circuits can behave as winner-take-all mechanisms of variable strength and can achieve close to optimal signal discrimination in the presence of external noise. PMID- 12079552 TI - CCCP algorithms to minimize the Bethe and Kikuchi free energies: convergent alternatives to belief propagation. AB - This article introduces a class of discrete iterative algorithms that are provably convergent alternatives to belief propagation (BP) and generalized belief propagation (GBP). Our work builds on recent results by Yedidia, Freeman, and Weiss (2000), who showed that the fixed points of BP and GBP algorithms correspond to extrema of the Bethe and Kikuchi free energies, respectively. We obtain two algorithms by applying CCCP to the Bethe and Kikuchi free energies, respectively (CCCP is a procedure, introduced here, for obtaining discrete iterative algorithms by decomposing a cost function into a concave and a convex part). We implement our CCCP algorithms on two- and three-dimensional spin glasses and compare their results to BP and GBP. Our simulations show that the CCCP algorithms are stable and converge very quickly (the speed of CCCP is similar to that of BP and GBP). Unlike CCCP, BP will often not converge for these problems (GBP usually, but not always, converges). The results found by CCCP applied to the Bethe or Kikuchi free energies are equivalent, or slightly better than, those found by BP or GBP, respectively (when BP and GBP converge). Note that for these, and other problems, BP and GBP give very accurate results (see Yedidia et al., 2000), and failure to converge is their major error mode. Finally, we point out that our algorithms have a large range of inference and learning applications. PMID- 12079553 TI - Fast curvature matrix-vector products for second-order gradient descent. AB - We propose a generic method for iteratively approximating various second-order gradient steps - Newton, Gauss-Newton, Levenberg-Marquardt, and natural gradient in linear time per iteration, using special curvature matrix-vector products that can be computed in O(n). Two recent acceleration techniques for on-line learning, matrix momentum and stochastic meta-descent (SMD), implement this approach. Since both were originally derived by very different routes, this offers fresh insight into their operation, resulting in further improvements to SMD. PMID- 12079554 TI - Representation and extrapolation in multilayer perceptrons. AB - To give an adequate explanation of cognition and perform certain practical tasks, connectionist systems must be able to extrapolate. This work explores the relationship between input representation and extrapolation, using simulations of multilayer perceptrons trained to model the identity function. It has been discovered that representation has a marked effect on extrapolation. PMID- 12079555 TI - Using noise to compute error surfaces in connectionist networks: a novel means of reducing catastrophic forgetting. AB - In error-driven distributed feedforward networks, new information typically interferes, sometimes severely, with previously learned information. We show how noise can be used to approximate the error surface of previously learned information. By combining this approximated error surface with the error surface associated with the new information to be learned, the network's retention of previously learned items can be improved and catastrophic interference significantly reduced. Further, we show that the noise-generated error surface is produced using only first-derivative information and without recourse to any explicit error information. PMID- 12079556 TI - Design and validation of an enzyme-linked immunospot assay for use in clinical trials of candidate HIV vaccines. AB - The enzyme-linked immunosorbent (ELISPOT) assay, which enumerates peripheral blood mononuclear cells (PBMCs) releasing interferon gamma (IFN-gamma) on specific antigen stimulation, is becoming the assay of choice for evaluation of vaccine-induced cell-mediated immune responses in many clinical trials. A properly conducted trial requires the assays to be validated, especially should the trial lead to vaccine licensure. Here, the design and validation of an ELISPOT assay are described for use in clinical trials of candidate human immunodeficiency virus (HIV) vaccines, using a particular immunogen termed HIVA. This assay employs eight pools of 20 to 23 peptides each: seven pools are derived from the immunogen and one pool is derived from cytotoxic T cell epitopes of common human viruses serving as an internal positive control. The validation determined that first, the overall variation of a positive response of approximately 500 spot-forming units (SFU)/10(6) cells was 21%, while second, the average of 5 SFU/10(6) cells was detected for the seven HIVA-derived pools in HIV uninfected individuals; third, a positive response to a peptide added to the assay pools was not occluded by the other pool peptides; fourth, the frequencies detected in fresh PBMCs were 2- to 3-fold higher compared with the same samples that had been cryopreserved; and finally, all seven HIV-derived pools induced IFN gamma responses in PBMCs isolated from HIV-infected individuals. The limits of the validation of assays involving biological responses of living cells are discussed. PMID- 12079557 TI - Inhibition of HIV replication and macrophage colony-stimulating factor production in human macrophages by antiretroviral agents. AB - Macrophage colony-stimulating factor (M-CSF) enhances the susceptibility of macrophages to infection with HIV-1, in part by increasing the expression of CD4 and CCR5. Human monocyte-derived macrophages (MDMs) infected in vitro with HIV-1 endogenously produce M-CSF, with kinetics paralleling virus replication, which can lead to enhanced spreading of the infection. AZT and ritonavir both inhibit HIV replication, but their impact on M-CSF production by HIV-infected human MDMs is unknown. The dose response and kinetics of virus replication in the presence of AZT and ritonavir were determined for HIV-infected MDMs from HIV-seronegative donors. Harvested supernatants were monitored for reverse transcriptase activity, M-CSF production, and HIV proteins. Our data suggest that threshold levels of HIV replication must occur before maximum M-CSF production is induced. Addition of AZT or ritonavir before or after establishment of productive HIV infection dramatically reduces virus replication and M-CSF production by human MDMs. However, ongoing virus replication and M-CSF production are slow to return to baseline levels after addition of AZT or ritonavir, suggesting that HIV replication and virion release from infected macrophages continue long after initiation of antiretroviral therapy. Our results suggest that, in human macrophages, HIV-1 replication and M-CSF production are inextricably linked, such that inhibition of one leads to a concomitant reduction of the other. Low-level HIV replication and M-CSF release during ongoing antiretroviral therapies may facilitate the survival and maintenance of infected macrophages and suggests that additional therapies targeting M-CSF may be critical for elimination of macrophage reservoirs. PMID- 12079558 TI - Quantitation of simian cytokine and beta-chemokine mRNAs, using real-time reverse transcriptase-polymerase chain reaction: variations in expression during chronic primate lentivirus infection. AB - Cytokines and beta-chemokines are important mediators of the immune system and are expressed in many infectious diseases. To study cytokine and beta-chemokine profiles during pathogenesis of lentiviral infection and progression to AIDS in rhesus macaques, we established new quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR) assays based on TaqMan chemistry. Using synthetic RNA standards, we quantified mRNAs of IL-2, IL-4, IL-6, IL-10, IL-12 p40, interferon gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), RANTES, macrophage inflammatory protein 1 alpha (MIP-1 alpha), and MIP-1 beta in unstimulated peripheral blood mononuclear cells (PBMCs) and lymph nodes from macaques chronically infected with SIV or SHIV. Viremic monkeys with decreased CD4(+) T cell counts (<500 cells/microl) had significantly higher IL-10 mRNA expression than uninfected controls, which parallels the findings in HIV-1 infected humans. In addition, MIP-1 alpha, MIP-1 beta, and RANTES mRNA expression increased in viremic monkeys with decreased CD4(+) T cell counts; gene expression was inversely correlated with CD4(+) T cell counts, but not viral load. The newly established quantitative real-time RT-PCR assays will allow the determination of cytokine and beta-chemokine patterns in rhesus macaques in studies of microbial pathogenesis or vaccine development. PMID- 12079559 TI - Characterization of an HIV type 1 strain with preferential replication in adherent cells. AB - HIV-E, emerging from persistently infected HeLa-T4 cells, replicates better in fibroblasts and epithelial cells with respect to the parental, T cell-derived HIV T. The two viruses share the same env V3 loop, but differ in cellular molecules incorporated on the envelope. Even when similar amounts of virus attachment occurred, HIV-E replicated better than HIV-T in cells from solid tissues, and the response to exogenous Tat was more efficient. This might be related to the long terminal repeat (LTR), because HIV-E has a TAR duplication, and a mutation in the Sp1-II binding site. Epithelial cells deserve further study, because they may be important in vivo for variant selection and latency. PMID- 12079560 TI - Interleukin 1 beta and interleukin 6 potentiate retinoic acid-mediated repression of human immunodeficiency virus type 1 replication in macrophages. AB - All-trans-retinoic acid (RA) has been shown either to activate or repress human immunodeficiency virus type 1 (HIV-1) replication in primary monocyte-derived macrophages (MDMs). We systematically investigated the contribution that cell donor and virus differences make to this variability. We found that the effect of RA was cell donor dependent. In addition, the ability of RA to repress HIV-1 replication varied between different virus stocks. In no case did RA affect either virus entry or integration but instead affected the accumulation of viral mRNAs in infected cells. Despite the complex variability in RA responsiveness in untreated cells, we found that RA consistently repressed virus replication when the MDMs were treated with concentrations of interleukin 1 beta (IL-1 beta) and IL-6 that are expected at local sites of infection, where HIV-1-infected macrophages reside in vivo. PMID- 12079561 TI - Lack of evidence for HIV type 1-related SIVcpz infection in captive and wild chimpanzees (Pan troglodytes verus) in West Africa. AB - Serum from 387 chimpanzees (Pan troglodytes verus), caught in the wild or bred in captivity, was tested for antibody to HIV-1 and HIV-2, using second- and third generation enzyme immunoassays. Six samples were repeatedly positive; however, only one of these was Western blot positive. Serial sera drawn before and after the Western blot-positive samples were seronegative, and thus we conclude that this sample represented specimen contamination, or mislabeling. Thus, none of the 387 Pan troglodytes verus from West Africa were spontaneously infected with SIVcpz. Chimpanzees are known to be exquisitely susceptible to infection with HIV 1 when experimentally inoculated, and thus our findings suggest that HIV-1 related viruses do not exist in Pan troglodytes verus in the wild. As it has been convincingly shown that SIVcpz exists in wild Pan troglodytes troglodytes in Central Africa, this suggests that HIV-1 arose in Central Africa, but not in West Africa. PMID- 12079562 TI - Effect of etanercept (Enbrel) on interleukin 6, tumor necrosis factor alpha, and markers of immune activation in HIV-infected subjects receiving interleukin 2. AB - The effect of etanercept, a soluble p75 tumor necrosis factor (TNF) receptor:Fc fusion protein (Enbrel; Immunex, Seattle, WA) on plasma cytokines was evaluated in 11 HIV-infected subjects receiving highly active antiretroviral therapy (HAART) for 28 weeks with or without subcutaneous or intravenous recombinant human interleukin 2 (rhIL-2). Plasma IL-6 and C-reactive protein (CRP) levels increased after rhIL-2 treatment. Etanercept pretreatment attenuated these increases. Median plasma IL-6 levels were 20.29 pg/ml 4 days after rhIL-2 and 7.87 pg/ml 4 days after etanercept and rhIL-2 (p = 0.22); median CRP levels were 78.73 and 46.16 microg/ml, respectively (p = 0.03). An effect on TNF bioactivity could not be assessed as all measurements were below limits of detection. No significant changes were seen in temperature or plasma levels of IL-4, IL-10, IL 12, interferon gamma, or HIV-1 RNA levels. All subjects had undetectable or low level HIV-1 RNA levels before etanercept dosing. One subject died; however, her death was thought to be unrelated to etanercept. Pretreatment with etanercept may blunt activation of IL-6 and CRP expression induced by rhIL-2. The safety and utility of etanercept in HIV-infected persons should be explored further. PMID- 12079563 TI - Subtypes A, C, G, and recombinant HIV type 1 are circulating in Bangladesh. AB - We analyzed the genetic diversity of HIV-1 circulating in Bangladesh by direct sequencing and subsequent phylogenetic analysis of the V3 region of the env gene and p17 fragment of the gag gene from nine unrelated patients. The sequences from one sample grouped into subtype A, five samples grouped into subtype C, and one grouped into subtype G. In addition, two patients appeared to be infected with different recombinant viruses consisting of subtype A and unclassifiable viral sequences. Epidemiological analysis revealed heterosexual transmission in the majority of cases. Furthermore, most subjects had a history of traveling, either to India or to the Arabian Peninsula. This study shows that several HIV-1 subtypes are circulating in Bangladesh, and we conclude that there must have been several introductions of HIV-1 into the Bangladeshi population. PMID- 12079564 TI - Full-length HERV-H elements with env SU open reading frames in the human genome. AB - Human endogenous retroviruses (HERVs) are estimated to represent at least 1% of the human genome. An HERV-H env SU sequence (HERV-H19) was used to screen the high-throughput (htgs) and nonredundant (nr) databases for other HERV-H SU open reading frames (ORFs) and thus possible functional proteins. Using PCR with primers derived from HERV-H19 SU, we also obtained several new sequences with ORFs from a human DNA sample. In a phylogenetic analysis, ORF-containing sequences clustered with HERV-H sequences from chromosomes 1 and 2. SU ORF- and non-SU ORF-containing elements had about the same difference between 5' and 3' long terminal repeats (LTRs) (about 4%), indicating a similar time of integration. SU ORF sequences had a moderately high number of synonymous-versus nonsynonymous mutations, which indicates a selection for maintenance of the HERV H SU ORFs. PMID- 12079565 TI - Genetic variation of the protease and reverse transcriptase genes in HIV-1 CRF04_cpx strains. AB - To gain information about the genetic variation of the protease and reverse transcriptase gene-coding regions in HIV-1 strains belonging to CRF04_cpx, genotyping and drug susceptibility testing were performed on serum samples derived from seven patients carrying sequences belonging to this circulating recombinant form. Substitutions classically associated with resistance to antiretroviral drugs were observed in six of seven samples, including G48V, V82A, L90M, M46I in the protease protein, and K70R, D69D/N, M184V, T215F, K103N in the reverse transcriptase protein. Genotypic resistance patterns of CRF04_cpx samples were found to be similar to those identified in subtype B viruses, suggesting that the drug-selective pressure has similar effects on both subtype B and CRF04_cpx. PMID- 12079566 TI - HIV-1 Subtype A, D, G, AG and unclassified sequences identified in South Africa. AB - HIV-1 subtype C accounts for the vast majority of infections in South Africa. However, increasingly non-C subtypes are being detected. Here we report 10 viruses that contain sequences that group with subtypes A, D, and G as well as CRF02_AG and 1 that could not be classified. Most of these individuals were from other countries in Africa. Some of these sequences were in combination with subtype C, possibly indicating local recombination events. Although there is no indication of endemic spread of these viruses, continued monitoring is warranted to track genetic changes, which may impact on diagnostic testing, therapeutic responses to antiretroviral therapies, and vaccine design. PMID- 12079568 TI - Drop-out and continuity of care for patients suffering from schizophrenia and schizophrenia-like psychosis evaluated by means of routine registration. AB - For most patients suffering from schizophrenia or other chronic psychoses, uninterrupted contact with psychiatric services for a long period is necessary. By means of routine registrations in Copenhagen County, the use of services in 1995 for patients suffering from schizophrenia or other ICD-10 F2 diagnoses (n = 1356) was analysed. Substantial interregional differences were found in admission to more than one psychiatric department, admission to more than one similar ward, interruption in treatment and loss of contact with the patient. More than 25% had contact with at least four treatment modalities during 1995, and the proportion of patients who experienced interruption in treatment at least once during the year varied between 19% and 37%. It is concluded that routine registration of psychiatric services is a suitable means for quality assurance and ought to be used regularly in analyses like the one presented here. In concert with the literature, it is concluded that the interregional differences indicate that compliance with treatment can be improved by better organization of the psychiatric treatment. PMID- 12079569 TI - The effect of a biological explanation on attitudes towards homosexual persons. A Swedish national sample study. AB - Studies assessing attitudes towards homosexuals have revealed widespread homophobia. The first aim of this study was to assess potential changes in public attitudes after important legislative changes related to homosexuals in Sweden. The second aim was to test whether the attitudes differ: 1) between people who believe in biological vs. people who believe in psychological theories in explanation of homosexuality, 2) between men and women, and 3) between the older and younger age groups. To this end, a questionnaire survey of a representative, randomly selected, national sample of 992 adult Swedish residents was carried out. The response rate was 67%, which is considered high in this context. In conclusion, this study demonstrates a clear-cut change towards more tolerant attitudes towards homosexual men and women compared to earlier studies. The reasons for this change is discussed; among factors of importance are anti discrimination legislation, increased visibility of homosexual people, and that more people currently regard homosexuality as a biologically determined, natural variant of human sexuality than was the case 10 years ago. In accordance, this study gave further support to the notion that those who believe that homosexuality is caused by biological factors have a less restrictive view on homosexuality than do people who hold a psychological view. PMID- 12079570 TI - Effect of major depression on cognitive performance among treatment-seeking adolescents. AB - We investigated the association between cognitive performance and major depressive disorder (MDD) in adolescents seeking outpatient treatment. We used several tests comparing cognitive capacities between 16 adolescents with MDD and 25 adolescents not diagnosed as suffering from psychiatric illness according to the Structured Clinical Interview for DSM-III-R (SCID). No difference in neuropsychological test performance was found between adolescents with MDD and those without psychiatric diagnosis. Logistic regression analysis showed the only independent factor associated with MDD to be Emotional Tone (S2; odds ratio 1.13). The specific effects of MDD on the adolescents were restricted to the emotional area and were expressed as a broad deterioration in psychosocial functioning. PMID- 12079571 TI - Panic disorder and the Defence Mechanism Test. AB - The aim was to study defence categories according to the modified version of the Defence Mechanism Test (DMTm) and to see if there was a relationship between DMTm and severity of illness. The material consists of 23 patients with panic disorder according to DSM-III-R who participated in a long-term follow-up of two clinical trials. The most common defence categories were repression, denial, disavowal or denial of the threat relation or of the identity of the peripheral person. The patients with denial or polymorphous identification had more severe symptoms and the latter group also were more handicapped by their symptoms. Denial and disavowal or denial of the threat relation may be defence categories, which are not so effective in preventing the individual from experiencing anxiety. Polymorphous identification, although not so common, does not seem to be an appropriate defence among patients with panic disorder. PMID- 12079572 TI - A study of the Alzheimer's Disease Assessment Scale-Cognitive (ADAS-Cog) in an Icelandic elderly population. AB - The Alzheimer's Disease Assessment Scale (ADAS) is designed for screening of cognitive and non-cognitive dysfunctions characteristic of persons with probable Alzheimer's disease (AD). The cognitive part of the scale (ADAS-Cog) is both convenient for screening of probable AD and as a measure of cognitive functioning during drug intervention. The aim of this study was to translate the ADAS Cognitive sub-test (ADAS-Cog) into Icelandic and to study its application in an elderly Icelandic population. The Mini-Mental State Examination (MMSE) and the ADAS-Cog were administered to 20 AD patients and 20 controls. Each patient was also rated on the Global Deterioration Scale (GDS). The probable AD patients were divided into two groups based on their GDS: 3-4 and 5-6 points. The patients were also divided into two groups based on their MMSE score: very mild to mild (23-30 points) and mild to moderate (15-22 points). Furthermore, the subjects were divided into two age groups: 65-76 and 77-92 years. Results revealed a highly significant difference on MMSE (22.3 +/- 3.4; 26.8 +/- 1.6; P < 0.05) and ADAS Cog (18.4 +/- 7.7; 7.3 +/- 3.5; P < 0.05) scores for patients and controls respectively. AD patients also performed significantly worse than the elderly control group on eight of the 11 sub-tests. Thus, the present findings are mainly in line with those of previous studies. The scale exceeds other screening tests such as the MMSE in that it addresses in more detail the symptoms of AD and is valuable for early detection of the illness and staging. ADAS-Cog plays an important role in the diagnostic makeup of AD along with other detailed investigations, such as neuropsychological assessment. PMID- 12079573 TI - Incidents of violence in psychiatric inpatient care. AB - During a period of 18 months a systematic inventory of aggressive incidents was made on two psychosis wards at the psychiatric department of Danderyd Hospital, Stockholm, Sweden. The Staff Observation Aggression Scale (SOAS) for recording and assessing aggressive incidents was used. Staff subjected to violence or threat of violence was interviewed using a semi-structured questionnaire. In one part of the study, a somewhat reduced variant of the same questionnaire was used but this time staff answered anonymously. Aggressive incidents were distributed over the 24 h, but had two distinct peaks - morning and evening. Aggressive incidents most often occur with patients with some kind of a psychotic diagnosis, most often schizophrenic psychoses. The female staff was more exposed to violence than male. Female patients are also more violent to female staff than to male staff. There is a significant difference in the emotional reactions of female and male staff after aggressions. In the staff, men are more often frightened while women tend to get surprised. Still, staff members of both genders feel insulted and angry. The "research" interviews were experienced as therapeutic discussions by the staff. PMID- 12079574 TI - Sexually abused women after multimodal group therapy: a long-term follow-up study. AB - This study reports a long-term follow-up of 38 of 54 (70%) women who had participated in time-limited multimodal group treatment for the psychological sequelae of sexual abuse. The women had been highly symptomatic at the onset and were generally improved following treatment and at follow-up (M = 4.7, SD = 2.0 years). Nonetheless, many remained moderately symptomatic. Previous therapy and pre-therapy level of symptoms predicted a higher level of post-treatment symptoms, while better post-treatment status and younger age predicted fewer symptoms at the time of follow-up. Interpersonal functioning at follow-up was predicted by post-treatment interpersonal functioning. On the whole, clients perceived their interpersonal functioning as better at follow-up than it had been after treatment. Their use of any mental health services was modest in the follow up period. In general, the long-term follow-up status of these women was encouragingly positive. PMID- 12079575 TI - Statistical methodology in general psychiatric journals. AB - Development of psychiatric research methods requires systematic review of their status. This study describes the frequency with which various statistical research designs and methods are reported in general psychiatric journals. All original research articles in four psychiatric journals in 1996 were reviewed: The American Journal of Psychiatry (AJP), The Archives of General Psychiatry (AGP), The British Journal of Psychiatry (BJP) and the Nordic Journal of Psychiatry (NJP). Evaluation included 448 regular articles, which reported original research findings based on systematic collection and statistical analysis of research data. Aspects measured were research design, statistical methodology, description of procedures and presentation of results. Percentage frequencies of reported statistical procedures were compared between the journals. The policy of determining statistical significance was still the most generally used method of conducting research. Compared to earlier reviews, readers of psychiatric reports in 1996 more frequently encountered computer dependent multivariate techniques. There were differences in the utilization and reporting of statistical procedures among the journals. The authors of psychiatric journals might apply these results in designing their psychiatric research to present intelligible and compact analysis combined with a high quality presentation technique, thus being in line with the policy and presentation of the leading psychiatric journals. PMID- 12079576 TI - Cross-sectionally and longitudinally balanced effects of processing speed on intellectual abilities. AB - The processing speed theory of cognitive aging states that declines in intellectual abilities other than processing speed are mainly due to a slowing of processing speed. Cross-sectional studies have provided support for the processing speed theory as for age-related cognitive differences. Longitudinal studies, in turn, have provided weaker support for the processing speed theory as for cognitive age changes. The present study aims to reconcile this discrepancy of cross-sectional and longitudinal results by constructing a "fair" test of the processing speed theory, i.e., a test that balances the cross-sectional age range and the time period covered longitudinally. Data from 83 older adults came from the Bonn Longitudinal Study on Aging. Using hierarchical linear models, it is shown that, although cross-sectional age differences and longitudinal changes in the intellectual abilities were equal, processing speed only attenuated cross sectional age effects in cognition but virtually did not reduce longitudinal change effects in cognition. This persisting difference in the explanatory power of processing speed regarding age-related differences and age-related changes is discussed with reference to other longitudinal studies and statistical issues regarding cross-sectional and longitudinal studies of aging. PMID- 12079577 TI - Older adults filter irrelevant information during metaphor comprehension. AB - In general, older adults are less likely than younger adults to inhibit irrelevant information when reading literal text (Hasher & Zacks, 1988). Are older adults also less likely to inhibit irrelevant information during metaphor comprehension? Young (mean age 19.2 years) and older adults (mean age 73.6 years) participated in a timed property-verification task. Although the older adults were generally slower than the younger, they displayed the same pattern of data. After reading metaphors, both groups verified metaphor-relevant properties more quickly after a metaphor prime than after a literal control prime, whereas metaphor-irrelevant properties were responded to more slowly after metaphor primes, relative to the controls. These results suggest that older adults, like younger ones, enhance metaphor-relevant properties and filter out metaphor irrelevant properties during metaphor comprehension. Older adults' metaphor comprehension processes thus seem intact. Mechanisms for filtering out metaphor irrelevant information are considered. PMID- 12079578 TI - Cognitive aging and inhibitory efficiency in the Daneman and Carpenter's working memory task. AB - This study tests the hypothesis that the ability to inhibit already processed and actually irrelevant information influences performance in the reading span task (RST). French versions of the Stroop color-word task and of the Daneman and Carpenter's RST were administered to 151 participants from 30 to 80 years. In addition to the traditional span score, a score of vulnerability to intrusions was also computed as the number of intruding responses (words from preceding trials of the RST or nonfinal words). An analysis of variance showed a significant age effect on the reading span and on the resistance to interference, but no significant age effect on the vulnerability to intruding responses. A multiple-regression analysis was also made with the reading span score as the dependent variable, and with age, vulnerability to intrusions, and resistance to Stroop interference as independent variables. This analysis revealed that there was a relation between participants' vulnerability to intruding responses and their working memory span scores; the contribution of age and resistance to interference were very weak. In conclusion the present findings first support the idea that the working memory capacity undoubtedly involves some inhibitory control; however, because the participants' vulnerability to intruding responses is not clearly affected by age, the present findings also suggest that some part of age effects upon the working memory span has to be explained by another factor than a growing inefficiency in inhibitory control. PMID- 12079579 TI - Age differences in inhibition of schema-activated distractors. AB - In recent years a debate has ensued over the arguments that normal aging is associated with a reduction in the efficiency of inhibitory mechanisms underlying selective attention, leading to disruptions in the normal operation of working memory. The purpose of the study reported here was to investigate the relationship between age, inhibitory functioning, and working memory, adapting a procedure previously used by Gernsbacher and Faust (1991) to measure the suppression of activated, but no-longer-relevant information in working memory. Young and older adults viewed picture arrays of objects found in common settings, and then judged whether a target word named an item from the previously presented array. Older adults relative to young adults demonstrated less suppression of critical target items. The implications of these findings regarding inhibitory function in old age are discussed. PMID- 12079580 TI - Terminal change in cognitive function: an updated review of longitudinal studies. AB - We provide an update to Siegler's (1975) review of the terminal change literature. Articles identified through MEDLINE, CANCERLIT, PUBMED, and PSYCINFO from 1975 to 2000 were examined. Lower levels of cognitive function are related to increased risk of mortality; however, the relationship between rate of cognitive change and mortality is less clear. Although there has been an increase in the number of studies since our last review, prior studies have been limited by a lack of medical data, consideration of dementia, small sample sizes, and poor cognitive measures. The emphasis on Alzheimer's disease and the growth of epidemiology of aging in the past 25 years have provided well-characterized population studies with serial cognitive measures that provide an opportunity to test the theory of terminal change. PMID- 12079581 TI - Good news about the effects of bad old-age stereotypes. AB - The study investigates whether the confrontation with negative, age-stereotyped information leads to a deterioration of older adults' self-perceptions as suggested by the labeling theory, or whether older adults protect their self perceptions against deterioration as suggested by theories of the resilience of the aging self. Self perceptions, as well as peer-perception of other older adults in general were assessed in a pre-post design. In the experimental group, 60 older adults received negative information about competence in old age. The control group of 40 older adults received neutral information. In the experimental condition, the general perceptions of other older adults worsened whereas their self-perception improved. In the control condition, only small changes emerged. Furthermore, the change of self-perceptions varied by the perceived applicability of the negative information for oneself. We conclude that older adults use negative age stereotypes more as a standard of reference for downward comparison rather than integrating them into their self-perceptions. PMID- 12079582 TI - The effect of aging on the synchronization of human arm movements to external events. AB - The aim of this study was to analyze the synchronization of cyclical arm movements to an external event and to determine age-related differences when the coupling strength between perception and action is manipulated. A group of young and a group of older subjects had to track a moving light travelling horizontally while manipulating a lever in the same direction (in-phase) or in the opposite direction (anti-phase). The results showed a performance decrease for both age groups in the anti-phase condition as compared to the in-phase condition, yet this decrease was more pronounced for the older persons. Interestingly, this age effect disappeared with practice. Most likely the observed age-related performance decrement was due to the difficulties encountered by older people to correct on line movements when the visual information needed was not directly accessible. Nevertheless, this study also highlights the capacity of older people to regain the performance loss through training. PMID- 12079583 TI - Magnetically mediated hyperthermia: current status and future directions. AB - The use of hyperthermia in the treatment of cancers is appealing because, as a physical therapy, hyperthermia would have far fewer restrictive side effects than chemotherapy and radiotherapy, and it could be used in combination with these therapies. However, the currently available modalities of hyperthermia are often limited by their inability to selectively target tumour tissue and, hence, they carry a high risk of collateral organ damage or they deposit heat in a very localized manner which can result in under-treatment of a tumour. Magnetically mediated hyperthermia (MMH) has the potential to address these shortcomings. MMH consists of the localization of magnetic particles or seeds within tumour tissue followed by exposure to an externally applied alternating magnetic field to cause them to heat. Since this concept was introduced (over 40 years ago), MMH has evolved into four general sub-classes: arterial embolization hyperthermia (AEH), direct injection hyperthermia (DIH), intracellular hyperthermia (IH) and interstitial implant hyperthermia (IIH). It is the purpose of this article to review these four sub-classes in terms of experimental or clinical results, advantages, limitations and current status. PMID- 12079584 TI - Does local hyperthermia affect metastasis of a human prostate carcinoma grown in athymic nude mice? AB - PRIMARY OBJECTIVE: This study investigated whether local hyperthermia increases metastasis of prostate carcinoma xenografts in athymic nude mice. RESEARCH DESIGN AND METHODS: Human prostate carcinoma cells (1-LN-PC-3-1A) were injected into the animal's thigh. Mice were randomly divided into those treated with local hyperthermia (43 degrees C for 45 min, tumour volume 100 or 200 mm(3)) and untreated controls, and sacrificed when the tumour volume reached 500 mm(3). MAIN OUTCOMES AND RESULTS: Histological evaluation (hematoxylin and eosin staining) of lymph nodes showed metastases in 20/21 hyperthermia-treated and 17/21 control mice. Lymph node involvement, tumour weight at sacrifice, and tumour growth rate were not significantly different between groups. CONCLUSION: Single local hyperthermia treatment had no effect on lymph node metastatic spread in this model. PMID- 12079585 TI - Use of the vasodilator sodium nitroprusside to increase tumour temperature during loco-regional hyperthermia with a capacitive ring applicator in a rat tumour model. AB - The influence of sodium nitroprusside (SNP) induced hypotension (to a mean arterial pressure of 60 mmHg) on tumour and normal tissue temperature during hyperthermia (HT) was examined. Loco-regional HT was given to the calf of BD IX rats by external radiofrequency heating from a capacitive ring applicator. In experiments in rats with subcutaneous BT(4)An tumours, the mean tumour temperature increased by 0.49 degrees C from 42.36 to 42.85 degrees C, on average, during SNP-hypotension. This represented 58% of the increase in tumour temperature found in the same rats when the tumour circulation was stopped completely by sacrificing the rats. SNP-hypotension resulted in a decrease in mean muscle temperature from 41.73 to 41.23 degrees C. The temperature difference between the tumour and the underlying muscle thereby increased by approximately 1 degrees C, indicating that SNP can increase tumour temperature during HT without increasing the risk of heat-related damage to skeletal muscle. Experiments in rats without tumours were also done to further examine the effect of SNP hypotension on muscle temperature under different treatment conditions (variation of radiofrequency energy deposition and water bolus temperature). It was found that SNP decreased the muscle temperature during HT in two experiments where the average muscle temperature was 42.1 and 42.6 degrees C, respectively. In an experiment where the muscle temperature was 43.0 degrees C, on average, before SNP infusion, the muscle temperature increased during SNP-hypotension. This finding indicates that SNP-hypotension during HT may increase the risk of skeletal muscle necrosis with muscle temperatures at this level. PMID- 12079586 TI - Thermal enhancement of melphalan and oxaliplatin cytotoxicity in vitro. AB - It has been established that hyperthermia can enhance cytotoxicity of some chemotherapeutic agents. This has led to various clinical trials of thermochemotherapy, although many questions remain unanswered. The effects of various agents have been studied on animal tumours with different histopathology at elevated temperatures. These studies indicated that alkylating agents were most effective to all tumours at a moderately elevated temperature. Cisplatin was also effective to all tumours, but its effectiveness at 41.5 degrees C was less than that of alkylating agents. To quantitatively study these findings, the magnitude of thermal enhancement of melphalan, an alkylating agent, and that of oxaliplatin, a new platinum compound, were studied at 37-44.5 degrees C by the colony formation assay. The dose of each agent was kept constant, and cell survival was determined as a function of treatment time. The cell survival curve was exponentially related with treatment time at all test temperatures, and the T(0) (the time to reduce survival from 1 to 0.37) decreased with an increasing temperature. These results suggested that the cytotoxic effect of these agents occurred with a constant rate at 37 degrees C, and the rate was facilitated with an increasing temperature. This suggests that heat can accelerate the cytotoxic chemical reaction, leading to substantial thermal enhancement. The thermal enhancement ratio (TER, the ratio of the T(0) at 37 degrees C to the T(0) at an elevated temperature) increased with an increase in the temperature. The activation energy for melphalan at moderately elevated temperatures was largest among the agents tested in the laboratory and that for oxaliplatin was approximately half of the melphalan activation energy. This suggests that the thermal enhancement for the cytotoxicity of melphalan or alkylating agents might be the greatest. Potential mechanisms of thermal enhancement of cytotoxicity were discussed. PMID- 12079587 TI - Role of ATP in the sensitivity to heat and the induction of apoptosis in mammalian cells. AB - Heat-induced cell death and apoptosis were studied with respect to intracellular ATP. Studies on the relationship between hyperthermic cell-killing at 44 degrees C and cellular ATP levels in four cell lines grown as monolayers and six cell lines grown in suspension showed good correlations between cellular ATP levels and the sensitivity to heat. D(0) values (the dose required to reduce survival in the linear portion of the response by 63%) linearly increased with an increase in cellular ATP levels. No such changes in sensitivity to heat were observed between the cells cultured at different cell densities, regardless of the change in the cellular ATP level. These results suggest that cellular intrinsic ability to supply ATP rather than the level of pooled ATP per se is responsible for the thermal response. Heat-induced apoptosis in L5178Y cells was observed following treatment at 42 degrees C for 70 min, 44 degrees C for 20 min or 47 degrees C for 3 min, which corresponded to surviving fractions of 25, 0.6 and 0.8%, respectively, but not at 47 degrees C for 20 min, indicating that mild heat shock induced apoptosis. 2-deoxyglucose (2DG) and 2,4-dinitrophenol (DNP) increased the sensitivity to heat and affected the mode of cell death. Cells treated with 2DG and DNP (2DG/DNP) were heated at 42 degrees C for 20 min, and then incubated at 37 degrees C for up to 2h in the presence or absence of 2DG/DNP. In the absence of 2DG/DNP, the cellular ATP level recovered to 76% of the control level and DNA ladder formation was observed, whereas in the presence of 2DG/DNP, the cellular ATP level was further decreased (3-7% of the control) and no DNA fragmentation was detected. These results suggest that the inhibition of ATP synthesis is closely associated with the enhancement of sensitivity to heat and that ATP is required for the induction of apoptosis. PMID- 12079588 TI - Heat shock induces centrosomal dysfunction, and causes non-apoptotic mitotic catastrophe in human tumour cells. AB - Normal human diploid cells and various human tumour cells were heat shocked at 43 degrees C for 2h and allowed to recover at 37 degrees C. It was found that heat shock treatment transiently disrupted the immunostaining of centrosomes, and no centrosome staining was detected in either normal or tumour cells 24h after heat shock. Staining recovered thereafter in normal cells, but in tumour cells abnormal centrosomes, multiple and minute centrosomes were induced. While normal cells were arrested in G1 and G2 after heat shock, significant numbers of mitotic cells with multiple poles appeared in tumour cells. Subsequently, cells with multiple micronuclei increased in tumour cells with time after heat shock. Although the nuclear morphology of these cells was similar to that of the apoptotic cells, no DNA ladder formation was observed up to 4 days after heat shock. Furthermore, an in situ assay failed to detect signals representative of apoptosis, indicating that apoptosis did not appear to be involved in heat shock induced cell death of human tumour cells. Instead, cell lethality was associated with mitotic catastrophe. PMID- 12079590 TI - Cybersurgery-an argument for enterprise liability. PMID- 12079589 TI - Hyperthermic pre-treatment protects rat IPC-81 leukaemia cells against heat- and hydrogen peroxide-induced apoptosis. AB - It is well known that hyperthermia causes a transient tolerance of cells to a second heat challenge (acquired thermotolerance). The present study addresses the question of whether hyperthermic pre-treatment also increases the tolerance against heat- and hydrogen peroxide-induced apoptosis in rat IPC-81 leukaemia cells. This cell line exhibits an aberrant heat shock response which is characterized by a lack of the inducible Hsp70 isoform, even under conditions of heat or hydrogen peroxide stress, while the constitutively expressed Hsc70 and the inducible isoform of hemoxygenase (HO-1) are strongly enhanced by heat stress (43.5 degrees C; 30 min). In spite of this Hsp70 deficiency, hyperthermic pre treatment protects IPC-81 leukaemia cells against apoptotic cell death induced by heat or hydrogen peroxide, but is less effective against necrosis induced by higher doses of the applied stressors. Addition of hydrogen peroxide (25 microM) enhances the amount of bax mRNA, while the level of bcl-2 mRNA remains unchanged. No increase of bax mRNA, in contrast, could be detected in heat shock-primed IPC 81 cells when treated with hydrogen peroxide after a 12h recovery. These results indicate that hyperthermic pre-treatment may exert its anti-apoptotic function not only by enhanced expression of constitutive as well as inducible HSPs but also by lowering the level of bax transcripts and thereby increasing the Bcl 2/Bax ratio. PMID- 12079591 TI - The involuntary civil commitment of mentally ill persons in the United States and Romania: a comparative analysis. PMID- 12079592 TI - Regulatory and legislative attempts at limiting medical resident work hours. PMID- 12079597 TI - From clinician to client: the lived experience of mental illness. AB - Psychiatric researchers and clinicians are quick to pay homage to the idea that mental illness has no respect for occupational attainments, social status, or academic degrees. Yet there is a considerable gap between accepting this idea solely on the basis of one's involvement in research or caregiving, versus first hand experience. In this paper, the author delves into his own experience of mental illness in order to deepen clinicians' understanding of the personal meaning of facing such a condition on a daily basis. Special attention is paid to the persistence of obstacles to effective treatment and care, obstacles due not so much to financial constraints as to a lack of enlightenment among those charged with improving the lives of persons with mental illness. PMID- 12079593 TI - Physician habit evidence in informed consent cases. PMID- 12079598 TI - The effect of auricular acupuncture on psychophysiological measures of cocaine craving. AB - This study used a randomized pretest/posttest control group design to test the effectiveness of auricular acupuncture interventions in diminishing psychological and physiological changes associated with cocaine craving in 30 treatment-seeking cocaine-dependent patients. The experimental group received the real auricular acupuncture intervention, insertion of needles into ear point locations specifically targeted for drug withdrawal. The control group received sham or placebo auricular acupuncture, insertion of needles into ear point locations not targeted for any specific therapeutic benefit. Psychological (Cocaine Craving Questionnaire - Now) and physiological (skin conductance activity) changes associated with cocaine craving were measured. Results showed no differences between the control and the experimental group in diminishing psychological and physiological measures associated with craving. There were differences from pre- to posttest on measures of psychological but not physiological craving for the combined experimental and control groups. PMID- 12079599 TI - Help-seeking behaviors by Korean immigrants for depression. AB - Underutilization of mental health services by Asian immigrants has been an ongoing concern for those attempting to provide accessible care for people suffering from mental illness. The author investigated Korean immigrants' help seeking behaviors for depression to understand their underutilization of mental health services. The study involved 6 focus group discussions and 24 in-depth interviews with 70 Korean immigrants in New York City. Prolonged care within family and traditional Asian practices led to a delay in seeking mental health services, keeping many Korean immigrants with depression out of the delivery system. The lack of interface between formal service providers and psychiatric service providers also caused delayed treatment. This study suggests the need for a comprehensive care model based on community education, linking mental health care with other services, and cultural brokering, as ways to connect population needs with mental health service delivery. PMID- 12079600 TI - Mental health case management in Canada: job description analyses. AB - The purpose of this study was to examine case managers' (CM) job position descriptions through content analysis. There were 29 case manager job position descriptions examined. The job descriptions could be grouped into four categories: (1) what the individuals should bring to the position, (2) what they did as CM, (3) to whom they were accountable, and (4) what was valued. Themes included focus on client as individual, focus on the external/system, or outer directed work, and the articulation point between the individual and broader system. The CM works directly and indirectly in the system and community through education, organizing, planning, and networking, as well as through provision of direct care to the individual client. The expectations of the positions are extensive. PMID- 12079601 TI - Simple reminiscence: a stress-adaptation model of the phenomenon. AB - The phenomenon of Simple Reminiscence may play an important role in the individual's ability to adapt to anxiety-provoking stressors across the life span. However, a clearly articulated model of the underlying psychodynamics of the phenomenon has not been developed. In this paper, a proposed model of the phenomenon of Simple Reminiscence is presented. The important components of the model-developmental issues, triggers, uses, processes, and outcomes-are interpreted within the context of Peplau's conceptualization of stress and stress adaptation. Implications of the model for future empirical investigations of Simple Reminiscence are discussed. PMID- 12079602 TI - Exploring the complexity of compliance in schizophrenia. AB - A large body of literature indicates that people diagnosed with schizophrenia are highly likely to not comply with their prescribed treatment regime at some stage during the illness process. Factors that indicate the risk of noncompliance have been the subject of considerable research over a number of years. This paper presents an extensive review of the research literature on the subject of compliance in schizophrenia. A number of factors have constituted the focus of research into this area. These include: socio-demographic characteristics, including age, gender and socioeconomic status; illness factors including insight, psychiatric symptomatology, duration of illness, substance abuse, and adverse side-effects of medication; psychosocial factors such as health beliefs and social supports; and treatment factors including the nature of the therapeutic relationship between patients and health care professionals. While the results of relevant research do not provide a clear and conclusive picture of compliance, they provide important information to guide the pivotal role of the mental health nurse in facilitating patient compliance. PMID- 12079603 TI - Telemedicine versus in-person dermatology referrals: an analysis of case complexity. AB - The goal of this study was to determine whether teledermatology referrals differ significantly from in-person referrals with respect to case complexity and diagnosis of cases referred. Teledermatology cases were compared to in-person cases seen by the same university dermatologist who also reviews the teledermatology cases. These were also compared with in-person cases evaluated by a different dermatologist at local clinics using traditional referral patterns. Study parameters included Current Procedural Terminology (CPT) codes as a measure of case complexity, International Classification of Disease (ICD) codes as a measure of case types, and time from referral to actual consultation. The most common CPT codes used for teledermatology were 99241 and 99242 with no significant differences in the frequency of assigned CPT codes for teledermatology versus in-person consultation. An analysis of the diagnostic codes revealed no significant differences between the types of cases referred to telemedicine and those referred for in-person consultation. Time between referral and actual encounter with the dermatologist was significantly shorter via telemedicine than either local or university clinic in-person visits. PMID- 12079604 TI - International concerted action on collaboration in telemedicine: recommendations of the G-8 Global Healthcare Applications Subproject-4. AB - The main objectives of the G-8 Global Healthcare Applications Subproject-4 (G-8 GHAP-SP-4) were to establish an international concerted action on collaboration in telemedicine, telehealth, and health telematics (hereafter referred in this paper as telemedicine). In order to promote and facilitate the implementation of telemedicine or health telematics networks around the world, it was considered necessary to address certain key issues. Five thematic solution-seeking forums were held between May 1998 and December 1999. Each addressed a key issue, including interoperability of telemedicine and telehealth systems, impact of telemedicine on health care management, evaluation and cost effectiveness of telemedicine, clinical and technical quality and standards, and medico-legal aspects of national and international applications. The main objectives of these forums were to establish best practices and a thorough review of the issues and discussions among experts to determine the best solutions for the facilitation of global international telemedicine networks. More than 650 invited participants from 16 countries attended the five forums, which were of 2-3 days in duration. These forums provided a foundation for the exchange of ideas resulting in the initiation of collaborative activities. Based on these deliberations, a series of 21 recommendations were prepared by the national representatives of the G-8 GHAP SP-4. These recommendations propose to political leaders and health care managers of the G-8 and other countries roadmaps to follow in order to accelerate the achievement of a Global Society of Healthcare via Telemedicine, Telehealth, and Health Telematics. The 21 recommendations are presented in this report. PMID- 12079605 TI - Practical telepathology using a digital camera and the internet. AB - Digital camera technology has developed rapidly and a large choice of reasonably priced, user-oriented models are now available. These can be used for both macroscopic and microscopic photography with good resolution. Internet transmission of digital images also makes it possible to consult pathologists anywhere in the world. This study tests a simple, fast, and inexpensive method for practical transmission of images for diagnosis using a digital camera and the Internet. Using a commercial digital camera mounted with a phototube adapter to a light microscope (6 images per case on average), 2210 digital images (310 Mb) from 347 cases of gastrointestinal, lung, and uterus specimens were captured. Each image, stored in medium compression JPEG (Joint Photographers Experts Group) format with 1024 x 768 pixel resolution, required approximately 5 seconds to capture after the case had been reviewed and appropriate fields for imaging selected (30 seconds per case on average). The images were transmitted from Samsung Medical Center, Seoul, to Korea University Hospital, Seoul, and John Hunter Hospital, Newcastle, Australia. Transmission was 100% successful with a total upload time of 3 hours for 310 MB of data (31 seconds per case on average). The images were downloaded in 2 hours and viewed on a 17-inch color monitor with a maximal resolution of 1280 x 1024 pixels. Telepathology diagnoses were made with 95% and 97% concurrence by two pathologists at Korea University Hospital and John Hunter Hospital, respectively. We suggest that the current level of commercial technology yields fast, convenient and economical tools for practical telepathology diagnosis. PMID- 12079609 TI - Induction of oxidative stress in the tissues of rats after chronic exposure to TCDD, 2,3,4,7,8-pentachlorodibenzofuran, and 3,3',4,4',5-pentachlorobiphenyl. AB - The abilities of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,4,7,8 pentachlorodibenzofuran (PeCDF), 3,3',4,4',5-pentachlorobiphenyl (PCB126), and mixtures of these xenobiotics (toxic equivalents, TEQs) to induce oxidative stress in hepatic and brain tissues of rats have been investigated after chronic (30 wk) exposure to these congeners. TCDD, PeCDF, PCB126, and TEQs were administered daily to groups of rats at doses that corresponded to their toxic equivalency factors (TEFs), and the biomarkers of oxidative stress, including the production of superoxide anion, lipid peroxidation, and DNA single-strand breaks (SSBs), were determined in hepatic and brain tissues at the end of the exposure period. The three chemicals caused similar dose-dependent increases in the production of superoxide anion, lipid peroxidation, and DNA SSBs, which plateaued at certain dose ranges, followed by secondary increases at the higher dose levels. Similar effects were also produced by the TEQs; however, the dose dependent increases in the biomarkers of oxidative stress were continuous and never achieved plateau levels. Except for PCB126, where statistical analyses revealed greater productions of superoxide anion and lipid peroxidation in brain tissues as compared with hepatic tissues, no significant differences were revealed between the two tissues in response to the other xenobiotics or the TEQs. Nonsignificant differences were also revealed when comparing the effects induced by the TEQs with those induced by the individual chemicals. PMID- 12079610 TI - Nickel-induced plasma lipid peroxidation and effect of antioxidants in human blood: involvement hydroxyl radical formation and depletion of alpha-tocopherol. AB - To provide evidence for the oxidative effect of nickel (Ni) treatment on blood, lipid peroxidation (LPO) and hydroxyl radical (*OH) generation were examined in human plasma. Nickel chloride induced LPO in plasma of human blood in vitro in a concentration-dependent (0-10 mM) and time-dependent (0-2 h) manner. The *OH production in plasma was quantified by measurement of conversion of salicylic acid (SA) into its hydroxylated products, 2,3- and 2,5-dihydroxybenzoate (DHB). The concentrations of 2,3- and 2,5-DHB in plasma increased in a concentration dependent manner after Ni treatment for 1 h. Furthermore, a decreasing trend in alpha-tocopherol levels in plasma was observed after Ni treatment. Concurrent incubation with gluthathione (GSH), catechin (CTCH), and mannitol decreased lipid peroxidation and reduced *OH formation induced by Ni, but exacerbation of the decrease of alpha-tocopherol in plasma occurred with catechin. PMID- 12079612 TI - Toxicokinetics of 2-methylimidazole in male and female F344 rats. AB - The toxicokinetics of 2-methylimidazole (2-MI) were studied in male and female Fischer 344 rats after a single iv dose of 10 mg/kg or gavage dose of 25, 50, or 100 mg/kg. The 2-MI was formulated in 0.05 M phosphate-buffered saline (pH 7.4). The iv profiles could be best described by a two-compartment model with first order elimination. The terminal elimination half-life, volume of distribution at steady state, and clearance values were 0.78 and 0.85 h(-1), 1.5 and 1.9 L, and 4.97 and 12.0 L/h/kg for males and females, respectively. After a gavage dose, the plasma concentration time profiles could be best described by a one compartment model, no lag phase, and first-order absorption and elimination. The peak 2-MI plasma concentrations increased proportionately with dose and were reached within 35 to 50 min (T(max)) for all groups. The estimated half-life value for 2-MI was about 1 h for the iv group and the male 25-, 50-, or 100-mg/kg groups and female 25-mg/kg groups. Clearance increased for the male 100- and female 50- and 100- mg/kg groups. For a given dose group, clearance was also two to three times greater for female rats when compared to male rats. Absolute bioavailability for 2-MI was estimated to approach 97%. The results of this study indicated that 2-MI was (1) rapidly and completely absorbed, (2) quickly eliminated, (3) cleared differently for females than for males, (4) affected somewhat by dose for females, and (5) unlikely to undergo tissue accumulation following repeated exposure. PMID- 12079611 TI - Metabolism and cytotoxicity of aflatoxin b1 in cytochrome p-450-expressing human lung cells. AB - The mycotoxin aflatoxin B(1) (AFB(1)) is a hepatocarcinogen in many animal models and probably a human carcinogen. Besides being a dietary carcinogen, AFB(1) has been detected in dusts generated in the processing and transportation of AFB(1) contaminated products. Inhalation of grain dusts contaminated with AFB(1) may be a risk factor in human lung cancer. Aflatoxin B(1) requires cytochrome P-450 (CYP)-mediated activation to form cytotoxic and DNA-reactive intermediates, and this activation in human liver is mediated by the CYP 1A2 and 3A4 isoforms. Which isoforms are important in AFB(1) activation in human lung is not well understood. To investigate whether these CYPs can activate AFB(1) at low, environmentally relevant concentrations in human lung cells, SV40 immortalized human bronchial epithelial cells (BEAS-2B) that were transfected with cDNA for CYPs 3A4 (B3A4) or 1A2 (B-CMV1A2) were used. B-CMV1A2 cultured in 15 nM AFB(1) produced the AFB(1) glutathione conjugate (AFB(1)-GSH) and aflatoxin M(1) (AFM(1)), while B3A4 cells produced only aflatoxin Q(1) (AFQ(1)) at 0.15 microM AFB(1). Nontransfected BEAS 2B cells produced no metabolites, even at 1.5 mM AFB(1). Microsomes prepared from B-CMV1A2 and B3A4 cells activated AFB(1) to AFB(1) 8,9-epoxide (AFBO), while those from BEAS-2B cells did not produce AFBO. Cytosol from all three cell types was ineffective at glutathione S-transferase (GST)-mediated trapping of enzymatically generated AFB(1) 8,9-epoxide. B-CMV1A2 cells were 100-fold more sensitive to AFB(1) compared to B3A4 cells, and were 6000-fold more sensitive than control BEAS-2B cells. Western immunoblots confirmed that only B-CMV1A2 cells expressed CYP 1A2 protein, while CYP 3A4 was only in B3A4 cells. B-CMV1A2 cells were the most sensitive to AFB(1), followed by B3A4 cells. CYP 3A4, which has been predicted to activate AFB(1) primarily at higher AFB(1) concentrations, was also responsible for significant AFB(1) toxicity at low concentrations. These data indicate that human lung cells expressing these CYP isoforms are capable of activating AFB(1), even at environmentally relevant concentrations. PMID- 12079613 TI - Reproductive outcomes in the women of the Red River Valley of the north. I. The spouses of pesticide applicators: pregnancy loss, age at menarche, and exposures to pesticides. AB - In the current study, there was a modest but significant increase in risk (1.6- to 2-fold) for miscarriages and/or fetal loss occurring throughout the year in the spouses of applicators who use fungicides. There is a surprisingly significant deficit in the number of male children born to the spouses of fungicide applicators. First-trimester miscarriages occur most frequently in the spring, during the time when herbicides are applied. Use of sulfonylurea (odds ratio OR = 2.1), imidizolinone (OR = 2.6) containing herbicides, and the herbicide combination Cheyenne (OR = 2.9) by male applicators was statistically associated with increased miscarriage risk in the spring. Limited survey data from women who are the spouses of applicators did not show major alterations of long-term endocrinologic status (menarche, menopause, endometriosis). With regard to personal pesticide exposures, only women who engaged in pesticide application where there is direct exposure to these products are at demonstrable risk (OR = 1.8) for miscarriage. It was hypothesized that the overall reproductive toxicity observed in this population is, for the greater part, a male-mediated event. Clarification of exposure events leading to reproductive toxicity through direct measurements of exposure in both men and women is needed to resolve this issue. PMID- 12079614 TI - Temperature-modulated carcinogenicity of 7,12-dimethylbenz[a]anthracene in rainbow trout. AB - Temperature-modulated hepatic disposition, covalent binding of radiolabeled genotoxin to hepatic DNA, and cancer incidence in rainbow trout (Oncorhyncus mykiss) were assessed after a single exposure to 7,12-dimethylbenz[a]anthracene (DMBA). Fish (2 g) were acclimated at 10, 14, or 18 degrees C for 1 mo and then exposed to 1 ppm DMBA in their water for 20 h. Exposures were at respective acclimation temperatures, or 10 and 18 degrees C acclimated fish were shifted to 14 degrees C for DMBA exposures. After 4 but not 20 h of exposure, hepatic [(3)H]DMBA equivalents increased with temperature for fish exposed at their respective acclimation temperatures (10 or 18 degrees C). Covalent binding of [(3)H]DMBA to hepatic DNA was similar after 3 d in fish exposed at their respective acclimation temperatures. However, in fish exposed at 14 degrees C, after 3 d the concentration of [(3)H]DMBA covalently bound to hepatic DNA was higher in 10 degrees C than 18 degrees C acclimated fish. After 21 d, covalent binding of [(3)H]DMBA to hepatic DNA was less persistent in 18 degrees C than 10 degrees C acclimated, exposed, and reared fish. There were no differences between temperature-shifted groups at that time. Temperature effects on tumor incidence were assessed 9 mo after DMBA waterborne exposures in fish that were reared at (1) their respective acclimation and exposure temperatures, (2) 14 degrees C after exposure at their respective acclimation temperature, and (3) 14 degrees C after 14 degrees C exposures. Incidence of stomach, liver, and swimbladder cancer increased dramatically with rearing temperature. Differences in tumor incidence were less marked in fish reared at the same temperature (14 degrees C). A strong negative correlation between liver tumor incidence and persistence of [(3)H]DMBA equivalents covalently bound to hepatic DNA suggested increased error-prone DNA repair at warmer temperature played an important role in increased tumor incidence. PMID- 12079615 TI - Alterations of motor nerve functions in animals exposed to motorcycle exhaust. AB - The effect of motorcycle exhaust (ME) on the motor nerve was studied using animals exposed to the exhaust by inhalation, intratracheal, or intraperitoneal administration of ME particulate (MEP). A 4-wk ME inhalation and intratracheal instillation of MEP for 1 d in rats or intraperitoneal administration of MEP (0.5 g/kg/d for 1 d and 0.1 g/kg/d for 2 d) in mice significantly decreased both rota rod performance and motor nerve conduction velocity. The effect of some polycyclic aromatic hydrocarbons on the motor nerve was also investigated. Treatment with benzo[a]pyrene (0.05 and 0.1 g/kg/d for 3 d), but not pyrene (0.1 g/kg/d for 3 d), resulted in significant decrease of motor nerve conduction velocity in mice. Moreover, the Na(+),K(+)-ATPase activities of sciatic nerves isolated from ME-, MEP-, or benzo[a]pyrene-exposed animals were decreased. Treatment with pyrene did not markedly affect the Na(+),K(+)-ATPase activity of sciatic nerve. The rats exposed to ME for 4 wk showed increases in blood and sciatic nerve manganese levels. Results indicate that motorcycle exhaust produces adverse effects on the motor nerve, which is associated with a fall in nerve Na(+),K(+)-ATPase activity. PMID- 12079616 TI - Oxidative DNA damage in male Wistar rats exposed to di-n-butyl phthalate. AB - Dialkyl phthalate esters are used in the plastic industry and widely distributed in the environment. Previously, it has been shown that di-n-butyl phthalate (DBP) produces testicular atrophy and liver enlargement in rodents, and the mechanisms behind this could involve reactive oxygen species (ROS). In this study, oxidative DNA damage was measured in terms of the premutagenic modified nucleoside 7,8 dihydro-8-oxo-2'-deoxyguanosine (8-oxodG) in nuclear DNA from liver, kidneys, and testes from rats exposed to DBP in the perinatal or preadult period. In one experiment, pregnant rats were administered 0 or 0.5 g DBP/kg/d by gavage from d 7 after conception to d 17 after delivery and organs from male offspring were analyzed. In a second experiment, 25-d-old rats were administered 0, 0.5, or 2 g DBP/kg/d by gavage for 10 d. After perinatal exposure, body and organ weights were unchanged. The 8-oxodG/10(6) dG ratio in liver DNA increased significantly in the exposed group. In contrast, the 8-oxodG/10(6) dG ratio was significantly decreased in kidney DNA, whereas it remained unchanged in the testis. After preadult exposure (postnatal d 25 to 34) the testes weight of the exposed animals were significantly decreased and severe atrophy of the seminiferous tubules was observed. The body weight of the animals in the high-dose group was significantly decreased compared to the control. The 8-oxodG levels in liver, kidney, and testis DNA remained unchanged. Although ROS has been suspected of being involved in the formation of testicular atrophy in phthalate-exposed rats, no apparent sign of oxidative DNA damage was found after phthalate exposure perinatally or during the preadult stage. With respect to phthalate-induced oxidative DNA damage in the liver, it appears that the developmental stage during exposure is important. PMID- 12079617 TI - Lipid response to pioglitazone in diabetic patients: clinical observations from a retrospective chart review. AB - The objective of this study was to determine whether improvements in the lipid profile observed in controlled clinical trials with pioglitazone are seen in the clinical practice setting, and to ascertain the influence of concurrent statin treatment. Charts of 100 consecutive patients with type 2 diabetes (mean age 56.8 years) treated with pioglitazone (45 mg/day) for 2-4 months were retrospectively analyzed for changes in serum lipids, glycemic parameters, and body weight. Subanalyses were performed on the relationship of lipid changes to baseline lipid values and to concurrent statin therapy. Pioglitazone was associated with statistically significant (p < 0.001) changes from baseline in HbA(1C) (mean decrease 1.09%), body weight (mean increase 1.76 kg), HDL cholesterol (HDL-C) levels (mean increase 15.6%), and triglycerides (mean decrease 9.9%). There was an increase (+ 1.09%) in mean individual LDL-C levels from baseline values, but this change was not statistically significant. The greatest absolute and percentage improvements in HDL-C and triglycerides were observed in patients who had the greatest lipid abnormalities at baseline: in patients with baseline HDL-C < 35 mg/dL, mean individual HDL-C values increased by 31% (p < 0.001); in those with baseline triglycerides >399 mg/dL, triglyceride levels decreased by 46% (p < 0.001); and in patients with baseline LDL-C > 129 mg/dL, mean individual LDL-C values decreased by 10.6% (p < 0.001). Subgroup analysis showed similar beneficial changes in HDL-C and triglycerides in patients who were not receiving concurrent statin therapy (n = 48) as in those who were receiving statins (n = 49). This observational study demonstrated that significant improvements in HDL-C and triglyceride levels can be achieved with pioglitazone in the clinical practice setting. The greatest improvements occurred in patients with the worst baseline lipid levels, and benefits were seen regardless of whether patients were receiving concurrent statin therapy. PMID- 12079618 TI - Does hypertriglyceridemia present an indication for pioglitazone therapy in diabetes? PMID- 12079619 TI - Inhaled insulin and insulin antibodies: a new twist to an old debate. AB - The recent development of inhaled insulin for the treatment of diabetes has the potential to significantly improve patient compliance and diabetes control. Preliminary studies have shown inhaled insulin to be effective in lowering blood glucose and HbA(1c) levels. However, inhaled insulin may stimulate insulin antibody production more than is commonly observed with highly purified human insulin administered subcutaneously. The significance of insulin antibodies in patient care has been a topic of frequent debate and has been studied extensively. This review will discuss the potential implications of elevated insulin antibody levels, the role insulin antibodies play in the metabolic control of diabetes and the effect inhaled insulin may have on the immune system. PMID- 12079620 TI - Physiological and therapeutic roles of peroxisome proliferator-activated receptors. AB - The peroxisome proliferator-activated receptors (PPARs) are a group of nuclear receptor isoforms, including PPARgamma, PPARalpha, and PPARdelta, encoded by different genes. PPARs are ligand-regulated transcription factors that control gene expression by binding to specific response elements (PPREs) within promoters. PPARs bind as heterodimers with a retinoid X receptor and, upon binding agonist, interact with cofactors increasing the rate of transcription initiation. The PPARs play a critical physiological role as lipid sensors and regulators of lipid metabolism. Natural ligands for the PPARs include fatty acids and eicosanoids. More potent synthetic PPAR ligands, including the fibrates and thiazolidinediones, are effective in the treatment of dyslipidemia and diabetes. Use of selective ligands led to the discovery of additional potential roles for the PPARs in pathological states, including atherosclerosis, inflammation, and hypertension. This review provides an overview of the molecular mechanisms of PPAR action and the involvement of the PPARs in the etiology and treatment of several chronic diseases. PMID- 12079621 TI - Unresolved challenges with insulin therapy in type 1 and type 2 diabetes: potential benefit of replacing amylin, a second beta-cell hormone. AB - Current insulin therapy still fails to safely restore near-normoglycemia in the majority of patients. Among the barriers to achieving tight long-term glycemic control with insulin in both type 1 and type 2 diabetes are an increased risk of hypoglycemia, undesired weight gain, and a failure to normalize postprandial hyperglycemia and excessive unpredictable diurnal glucose fluctuations. Amylin is a second beta-cell hormone that is cosecreted with insulin in response to meals, and is deficient in patients with type 1 and insulin-requiring type 2 diabetes. Preclinical studies indicate that amylin acts as a neuroendocrine hormone that complements the effects of insulin in postprandial glucose regulation by suppressing postprandial glucagon secretion and slowing the rate of nutrient delivery from the stomach to the small intestine. Human amylin is not optimal for replacement therapy because of its propensity to aggregate; thus, pramlintide, a soluble, nonaggregating synthetic peptide analog of human amylin, was developed that has potency at least equal to that of human amylin. In clinical studies, subcutaneous injections of pramlintide prior to meals, in addition to insulin therapy, significantly reduced postprandial glucose excursions and lowered HbA(1c) levels in patients with type 1 and type 2 diabetes. The improvement in long-term glycemic control was associated with a significant reduction in body weight and occurred without increases in total daily insulin use or in overall severe hypoglycemia event rates. Because of this unique spectrum of clinical effects, amylin replacement with pramlintide as an adjunctive therapy to insulin is a promising approach that may fulfill some of the unmet clinical needs of insulin-using patients with type 1 and type 2 diabetes. PMID- 12079622 TI - Is pramlintide an adjunct to insulin therapy? PMID- 12079623 TI - A dietary cause of type 1 diabetes: unearthing a new twist to the tale. PMID- 12079624 TI - The First Annual Diabetes Technology Meeting--a new forum for accelerating the development of technology to fight diabetes. PMID- 12079625 TI - An overview of human factors engineering at CDRH in the safety and effectiveness of blood glucose meters. PMID- 12079626 TI - Oxidative phosphorylation: key to life. PMID- 12079627 TI - Comment on Lehmann and Tatti: proposed controlled trial on simulators in diabetes education. PMID- 12079630 TI - Non-ablative cutaneous remodeling with a 1.45 microm mid-infrared diode laser: phase I. AB - BACKGROUND: Presented here is phase I of a three-part study of non-ablative cutaneous remodeling with a 1.45 microm diode laser configured with a cryogen spray cooling device. METHODS: Eight men and two women (average age of 67 years) with Fitzpatrick skin phototypes I-IV were treated at postauricular sites. Treatment consisted of one or two passes with a 4mm spot and an average power of 12 W. Heating times ranged from 150 ms to 500 ms applied over two to six treatment cycles. One treatment 'cycle' lasted for 100 ms and consisted of programmable parallel cryogen spray cooling pulses interspersed with unopposed heating. Biopsies were obtained at baseline, immediately after treatment, and at 2 months. RESULTS: Patients were assessed 1 day, 1 week, 1 month and 2 months after treatment. Treatments were well tolerated with minimal pain, with a trend towards increasing discomfort with longer heating times. Erythema and edema were also mild and short-lived. There was a direct relationship between the degree of erythema and edema, and longer unopposed heating times. Mild hyperpigmentation occurred at only three treatment sites. Epidermal burns usually presented as immediate whitening in 11 of 60 one-pass sites and four of 16 two-pass sites. Whitening was associated with longer unopposed heating times. Atrophic, pitted scars occurred at two single-pass sites and three double-pass sites. Baseline biopsies demonstrated solar elastosis in a 375 microm thick band (mean range from 100 microm to 480 microm deep in the dermis). Immediate post-treatment biopsies demonstrated thermal damage in a 333 microm thick band (mean range from 311 microm to 644 microm deep in the dermis). Finally, dermal fibrosis was observed 2 months after treatment in a 272 microm thick band (mean range from 148 microm to 420 microm deep in the dermis). CONCLUSION: The 1.45 microm diode laser is capable of targeting dermal collagen and stimulating fibrosis at depths where solar elastosis resides. Longer unopposed heating times corresponded to increased erythema, edema, and pain, which were typically mild and short-lived. Epidermal burns can result in pitted scars. PMID- 12079631 TI - Non-ablative cutaneous remodeling with a 1.45 microm mid-infrared diode laser: phase II. AB - BACKGROUND: Presented here is phase II of a three-part study of non-ablative cutaneous remodeling with a 1.45 microm diode laser configured with cryogen spray cooling. In phase I, safe heating and cooling parameters were established by examining gross and microscopic changes induced by the laser. Phase II examines clinical changes and side effects in the treatment of single facial rhytids. METHODS: Two men and seven women with Fitzpatrick skin phototypes I-III were treated. Single facial rhytids were treated on three separate occasions 3 weeks apart (six periorbital and three perioral). Single, control wrinkles on the contralateral sides were treated with cryogen spray cooling alone. Subjects were treated with single passes with a 5mm spot for the first two treatments. Owing to a modification in the handpiece design, a 4mm spot was used for the third and final treatment. The average power was 12 W. At each treatment visit, heating times ranged from 200 ms to 300 ms, applied as a series of heating/cooling cycles. One treatment 'cycle' lasted for 100 ms and consisted of continuous laser heating interspersed with programmable parallel cryogen spray cooling bursts. RESULTS: Patients were assessed 1 day, 1 week, 4 months, and 6 months after treatment. Treatments were well tolerated, and no patient required pain control pre or post operatively. Mild erythema and edema were noted immediately after treatment and typically cleared within 2-3 days. Superficial, branny hyperpigmentation occurred in six patients at both treatment and control sides. This discoloration resolved within 1 week of treatment and left no residual pigment alterations. No whitening or residual scarring occurred. Rhytid scores improved from a baseline score of 2.3 to 1.8 at 6 months after treatment (p>0.05). Patient acceptance of the treatment was high, but most felt that there was little improvement of the treated rhytids. CONCLUSION: Although the 1.45 microm diode laser is capable of targeting dermal collagen and stimulating fibrosis at depths where solar elastosis resides, clinical improvement of rhytids was mild and did not correlate well with the degree of histologic changes noted in phase I. PMID- 12079632 TI - Botulinum toxins types A and B for brow furrows: preliminary experiences with type B toxin dosing. AB - BACKGROUND: Facial lines resulting from hyperactivity can be misleading manifestations of negative emotions, fatigue and stress. They may also contribute to a perception of facial aging. A well established treatment is botulinum toxin type A (BTX-A). Recently, botulinum toxin type B (BTX-B) has become available for the treatment of cervical dystonia. There has been little comparison on the efficacy of the two different types of botulinum toxins, nor is there information on appropriate dosing of BTX-B for facial muscles. OBJECTIVES: The purpose of this pilot study was to observe the effects of BTX-B in comparison to BTX-A, on patients with brow furrows assessing initial efficacy and duration of effect. METHODS: Patients were injected with BTX-B in two different dose conversions against BTX-A to the corrugator-procerus complex. Some patients received a conversion of 50 units of BTX-B (total of 1000 units) to one unit of BTX-A while others received a conversion of 100 units of BTX-B (total of 2000 units) to one unit of BTX-A. The patients treated with BTX-A received a total of 20 units. These patients were clinically assessed prior to treatment and 3 days, 1 week, 4 weeks, 12 weeks and 16 weeks after treatment. RESULTS: Both types of botulinum toxin were effective at improving glabellar frown lines. The onset of actions occurred slightly sooner (2-3 days) with BTX-B than with BTX-A (3-7 days). Duration of effect with BTX-A was at least 16 weeks. With 1000 units of BTX-B, dose duration was 6-8 weeks and with 2000 units of BTX-B, duration was 10-12 weeks. SUMMARY: Both types of botulinum toxin are effective at correcting deep glabellar furrows. At least with the doses used, BTX-B has a quicker onset of action and BTX-A has longer benefit for glabellar wrinkles. These data strongly suggest that further dose ranging studies of BTX-B are necessary and indicated in controlled double blind studies in a larger patient population. PMID- 12079633 TI - Botulinum B treatment of the glabellar and frontalis regions: a dose response analysis. AB - BACKGROUND: Botulinum toxin injections represent the most commonly performed cosmetic procedure in the US. There is an enormous reported experience documenting the efficacy of botulinum toxin A injections. There is very limited published information about the efficacy and appropriate dosage of botulinum toxin B injections. OBJECTIVE: The aim of this study was to evaluate the dosage response and side effect profile of botulinum toxin B injections. METHOD: Twenty six subjects received botulinum B injections to their glabellar area. Eighteen subjects received botulinum B injections to their frontalis region. Three different dosages were used. In the glabellar treated subjects the low dose group received a total of 1875 units; the medium dose group received a total of 2500 units; and the high dose group received a total of 3125 units. In the frontalis treated group, the low dose group received a total of 2250 units; the medium dose group received a total of 3000 units; and the high dose group received a total of 3750 units of botulium B toxin. RESULTS: In the glabellar group, most subjects showed some evidence of paralysis at 2 months. Only in the high dose group did a significant cadre of treated subjects still show a significant response at 3 months after treatment. In the frontalis group, response was often still seen at 2 months. However, at 3 months most subjects showed no continued effect from botulinum B toxin. CONCLUSION: Botulinum B toxin injections represent an alternative to botulinum A treatment for glabellar and frontalis hyperkinetic wrinkles. At the doses utilized in this study, the effect does not generally appear to last as long as has been reported with botulinum A injections. However, the onset of action may be sooner with botulinum B injections. PMID- 12079634 TI - Inotropic therapy of heart failure. Editorial comments on: Vasodilation and mechanoenergetic inefficiency dominates the effect of the "Ca2+ sensitizer" MCI 154 in intact pigs. PMID- 12079635 TI - Strategies for spinal cord protection during descending thoracic and thoracoabdominal aortic surgery: Up-to-date experimental and clinical results -- a review. PMID- 12079636 TI - Monophasic action potential mapping in swine and humans using modified-tip ablation catheter and electroanatomic mapping system. AB - OBJECTIVE: To evaluate the feasibility of monophasic action potential (MAP) mapping using a modified-tip NaviStar catheter in swine and humans. METHODS: MAP mapping was performed using the modified-tip catheter at 71 +/- 21 atrial and 60 +/- 16 ventricular sites in 10 healthy pigs and at 56 ventricular sites in one patient, and using an ordinary Navi-Star catheter at 30 atrial sites in one patient and 50 +/- 14 ventricular sites in four patients. In an additional 20 patients, MAPs were also recorded at 9 +/- 2 atrial sites using the modified-tip catheter or at 12 +/- 9 atrial sites using the ordinary catheter. RESULTS: In pigs, the plateau amplitudes of the MAPs recorded using the modified-tip catheter were 4.1 +/- 3.2 mV for the atrial and 9.5 +/- 4.3 mV for the ventricular MAPs. In patients, both the ventricular and atrial MAPs recorded using the modified-tip catheter were significantly higher than using the ordinary catheters, 15.7 +/- 8 and 3.0 +/- 0.9 mV vs 9.5 +/- 3.9 and 2.0 +/- 0.6 mV for the ventricular and atrial MAPs, respectively (p < 0.0001). The baseline disturbances were <10% of the MAP amplitude in 95% of the pig and 96% of the patient MAPs. CONCLUSION: A modified-tip Navi-Star catheter could be used in swine and in humans for prompt recording of MAPs with acceptable amplitudes and baselines. MAP mapping using the modified-tip catheter is safe and feasible for clinical use. PMID- 12079637 TI - Analysis of heart rate variability does not identify the patients at risk of atrial fibrillation after coronary artery bypass grafting. AB - OBJECTIVE: Analysis of heart rate variability (HRV) is a non-invasive method for assessing cardiac autonomic regulation. This study was designed to investigate whether preoperative short-term HRV analysis could identify the patients at high risk of atrial fibrillation (AF) after coronary artery bypass grafting (CABG). DESIGN: HRV measurement was performed preoperatively for 92 elective CABG patients. RESULTS: Thirty patients had AF postoperatively while 62 remained in sinus rhythm (SR). Logistic regression analysis identified two independent predictors for AF after CABG: increased age with an odds ratio (OR) 1.06 per year and higher body mass index (BMI) with an OR 1.18 per unit. However, no measured parameters of HRV analysis differed significantly between AF and SR groups. CONCLUSION: The patients at high risk of AF after CABG cannot be identified by preoperative short-term HRV analysis performed during standardized physiological conditions. Advanced age and higher BMI were independent risk factors of AF after CABG in this study. PMID- 12079638 TI - Vasodilation and mechanoenergetic inefficiency dominates the effect of the "Ca(2+)-sensitizer" MCI-154 in intact pigs. AB - OBJECTIVE: Ca(2+)-sensitizing agents hold potential as ideal cardiac inotropes, but effects in intact animals are scarcely described. We evaluated a pyridazinone derivative, MCI-154, for hemodynamic, inotropic, mechanoenergetic and oxidative metabolic effects. DESIGN: Intracavitary left ventricular (LV) pressure and conductance (volume) was assessed in open chest anesthetized pigs (n = 6). Contractile performance, pressure-volume area (PVA) and myocardial oxygen consumption (MVO(2)) were assessed. Myocardial substrate uptake and production of (14)CO(2 )(from glucose) and (3)H(2)O (from fatty acids) were monitored. MCI-154 administration: "low range": 0.1, 0.2, 0.3, 0.5 microg/kg/min and "high range": 0.75, 1.0, 2.0, 3.0 microg/kg/min. Parameters were compared with baseline and a time reference group (n = 7). RESULTS: MCI-154 induced a progressive dose dependent decrease in systemic vascular resistance, with a concomitant increase in heart rate and cardiac output. Contractility increased only in the high-dose range, and mechanoenergetic efficiency was significantly reduced by drug infusion in all doses. CONCLUSION: The pyridazinone derivative MCI-154 has minimal inotropic action, induces a significant "oxygen waste", and decreases vascular resistance in intact pigs. A potent phosphodiesterase inhibitory effect may explain this, which suggests further drug refinement. PMID- 12079639 TI - In vitro agreement between magnetic resonance imaging and intraluminal Doppler ultrasound for high flow velocity measurements. AB - OBJECTIVE: Differences of opinion in the literature about the agreement between magnetic resonance imaging (MRI) and ultrasound Doppler (USD) for high blood velocity measurement were the basis for this investigation. It compares in vitro velocity measurements by MRI with intraluminal USD, using a specially designed flow phantom. DESIGN: The flow phantom consisted of a pulsatile pump, tubing, a prosthetic heart valve, measurement sections and a reservoir. Peak systolic velocities (0.7-5.0 m/s, n = 10) were measured with standard MRI phase shift velocity mapping using different regions of interest (ROIs). Intravascular USD velocity measurements were carried out using a thin cannula measuring peak systolic local maximum and local mean velocity within the small sample volume. RESULTS: Using maximum velocity USD recordings, comparison with MRI velocities was poor when the ROI covered the complete vessel lumen giving offset -22.3% and limits of agreement (LOA) 57.5 and 105.1%. When the ROI was small the velocities were closer with offset -11.8% and LOA 75.3-103.3%. When the spatial resolution was comparable and the spatial mean was read from the USD recordings, the velocities showed good agreement with an offset close to zero (0.45%). CONCLUSION: This study shows the importance of selecting small ROIs when using MRI for evaluating valve stenosis. Only when using spatially mean velocities over comparable regions the two methods showed good agreement. PMID- 12079640 TI - Cardiomyocyte apoptosis after antiviral WIN 54954 treatment in murine coxsackievirus B3 myocarditis. AB - OBJECTIVE: Cardiomyocyte apoptosis (CA) is known to occur in experimental coxsackievirus B3 (CVB3) myocarditis. However, the mechanisms of CA induction are not well known. In this study we investigate the role of direct viral induction of CA in CVB3 myocarditis. DESIGN: A/J mice were infected with the Woodruff strain of CVB3 and treated with WIN 54954 for 5 days thereafter. WIN 54954, a compound that inhibits early events of picornavirus infection, is known to dramatically reduce mortality in murine CVB3 myocarditis without abrogating systemic or myocardial inflammation. Presence of viral RNA (in situ hybridization), CA (TUNEL method) and histopathology were studied in transverse ventricular sections at day 7 post infection (n = 8 treated and n = 8 non treated). RESULTS: The proportion of cardiomyocytes containing viral RNA was 89% lower in WIN 54954 treated mice when compared with non-treated mice (0.29 +/- 0.56% vs 2.76 +/- 1.65%, p = 0.003). Treatment also reduced the amount of CA by 52% compared with non-treated mice (0.20 +/- 0.06% vs 0.42 +/- 0.06%, p < 0.001). The reduction of CA by WIN treatment did not result in any increase of necrosis, in fact treatment reduced the area of necrotic lesions by 77% (2.51 +/- 1.64% vs 11.10 +/- 8.76%, p = 0.028). CONCLUSION: Taking the results of the reduced CA, necrosis and viral RNA with no effect on inflammation into account, our findings suggest the importance of direct viral effect in cardiomyocyte damage by both apoptosis and necrosis in CVB3 myocarditis. PMID- 12079641 TI - Classification of apocynaceae s.l. according to a new approach combining Linnaean and phylogenetic taxonomy. AB - A new approach to a nomenclatural system, including elements from both Linnaean and phylogenetic nomenclature, is proposed. It is compatible with the existing Linnaean system, including "standard names" corresponding to principal and secondary ranks, and uses a variant of the definitions from the Phylocode system. A new infrafamilial classification, using this nomenclatural approach, of the Apocynaceae s.l. (i.e., including the Asclepiadaceae) based mainly on analyses of rbcL and ndhF data is discussed. Twenty-one tribes and four rankless taxa are defined. PMID- 12079642 TI - Phylogenetic relationships in the genus mus, based on paternally, maternally, and biparentally inherited characters. AB - Several species in the rodent genus Mus are used as model research organisms, but comparative studies of these mice have been hampered by the lack of a well supported phylogeny. We used DNA sequences from six genes representing paternally, maternally, and biparentally inherited regions of the genome to infer phylogenetic relationships among 10 species of Mus commonly used in laboratory research. Our sample included seven species from the subgenus Mus; one species each from the subgenera Pyromys, Coelomys, and Nannomys; and representatives from three additional murine genera, which served as outgroups in the phylogenetic analyses. Although each of the six genes yielded a unique phylogeny, several clades were supported by four or more gene trees. Nodes that conflicted between trees were generally characterized by weak support for one or both of the alternative topologies, thus providing no compelling evidence that any individual gene, or part of the genome, was misleading with respect to the evolutionary history of these mice. Analysis of the combined data resulted in a fully resolved tree that strongly supports monophyly of the genus Mus, monophyly of the subgenus Mus, division of the subgenus Mus into Palearctic (M. musculus, M. macedonicus, M. spicilegus, and M. spretus) and Asian (M. cervicolor, M. cookii, and M. caroli) clades, monophyly of the house mice (M. m. musculus, "M. m. molossinus," M. m. castaneus, and M. m. domesticus), and a sister-group relationship between M. macedonicus and M. spicilegus. Other clades that were strongly supported by one or more gene partitions were not strongly supported by the combined data. This appears to reflect a localized homoplasy in one partition obscuring the phylogenetic signal from another, rather than differences in gene or genome histories. PMID- 12079643 TI - Phylogenetic placement of retropinnid fishes: data set incongruence can be reduced by using asymmetric character state transformation costs. AB - We used mitochondrial DNA sequences to determine the phylogenetic placement of southern smelts (Retropinnidae), a group of diadromous fishes endemic to New Zealand and Australia. Our genetic data strongly support a sister group relationship between retropinnids and northern hemisphere smelts (Osmeridae), a relationship that seems consistent with the similar appearance and life history strategies of these two groups. Our analysis indicates that Retropinnidae and Osmeridae together represent the sister group to the southern hemisphere galaxiid fishes (Galaxiidae). However, this finding conflicts with several recent osteological analyses, which supported a sister relationship for Retropinnidae and Galaxiidae, giving a monophyletic southern hemisphere assemblage (Galaxioidea). We review cases of incongruence and discuss factors that might explain significant disagreement between molecular and morphological data matrices. We suggest that repeated evolutionary simplification may have undermined the accuracy of morphological hypotheses of osmeroid relationships. Although equally weighted parsimony analysis of morphological data rejects the molecular hypothesis (Osmeridae + Retropinnidae), implementation of a range of weighting schemes suggests that incongruence is nonsignificant under asymmetric character transformation models. We propose that a simple "equal transformation cost" parsimony analysis may be biologically unrealistic, especially when reductive homoplasy is widespread; as is increasingly being accepted, complex character states are more readily lost than gained. Therefore, we recommend that morphological systematists routinely implement a range of character transformation models to assess the sensitivity of their phylogenetic reconstructions. We discuss the antitropical biogeography of osmeroid fishes in the context of vicariance and transequatorial dispersal. PMID- 12079644 TI - Robustness of ancestral state estimates: evolution of life history strategy in ichneumonoid parasitoids. AB - We test hypotheses for the evolution of a life history trait among a group of parasitoid wasps (Hymenoptera: Ichneumonoidea), namely, the transition among koinobiont parasitoids (parasitoids whose hosts continue development after oviposition) between attacking exposed hosts and attacking hosts that are concealed within plant tissue. Using a range of phylogeny estimates based on 28S rDNA sequences, we use maximum parsimony (MP) and maximum likelihood (ML) methods to estimate the ancestral life history traits for the main clades in which both traits occur (using the programs MacClade and Discrete, respectively). We also assess the robustness of these estimates; for MP, we use step matrices in PAUP* to find the minimum weight necessary to reverse estimates or make them ambiguous, and for ML, we measure the differences in likelihood after fixing the ancestral nodes at the alternative states. We also measure the robustness of the MP ancestral state estimate against uncertainties in the phylogeny estimate, manipulating the most-parsimonious tree in MacClade to find the shortest suboptimal tree in which the ancestral state estimate is reversed or made ambiguous. Using these methods, we find strong evidence supporting two transitions among koinobiont Ichneumonoidea: (1) to attacking exposed hosts in a clade consisting of the Helconinae and related subfamilies, and (2) the reverse transition in a clade consisting of the Euphorinae and related subfamilies. In exploring different methods of analyzing variable-length DNA sequences, we found that direct optimization with POY gave some clearly erroneous results that had a profound effect on the overall phylogeny estimate. We also discuss relationships within the superfamily and expand the Mesostoinae to include all the gall associated braconids that form the sister group of the Aphidiinae. PMID- 12079645 TI - Analyzing developmental sequences within a phylogenetic framework. AB - Heterochrony is important as a potential mechanism of evolutionary change. However, the analysis of developmental timing data within a phylogenetic framework to identify important shifts has proven difficult. In particular, analytical problems with sequence (event) heterochrony revolve around the lack of an absolute time frame in development to allow standardization of timing data across species. An important breakthrough in this regard is the method of "event pairing," which compares the relative timing of developmental events in a pairwise fashion. The resulting event-pair-encoded data can be mapped onto a phylogeny, which can provide important biological information. However, event paired data are cumbersome to work with and lack a rigorous quantitative framework under which to analyze them. Critically, the otherwise advantageous relativity of event-pairing prevents an assessment of whether one or both events in a single event-pair have changed position during evolutionary history. Building on the method of event-pairing, we describe a protocol whereby event pair transformations along a given branch are analyzed en bloc. Our method of "event-pair cracking" thereby allows developmental timing data to be analyzed quantitatively within a phylogenetic framework to infer key heterochronic shifts. We demonstrate the utility of event-pair cracking through a worked example and show how it provides a set of desired features identified by previous authors. PMID- 12079646 TI - An approximately unbiased test of phylogenetic tree selection. AB - An approximately unbiased (AU) test that uses a newly devised multiscale bootstrap technique was developed for general hypothesis testing of regions in an attempt to reduce test bias. It was applied to maximum-likelihood tree selection for obtaining the confidence set of trees. The AU test is based on the theory of Efron et al. (Proc. Natl. Acad. Sci. USA 93:13429-13434; 1996), but the new method provides higher-order accuracy yet simpler implementation. The AU test, like the Shimodaira-Hasegawa (SH) test, adjusts the selection bias overlooked in the standard use of the bootstrap probability and Kishino-Hasegawa tests. The selection bias comes from comparing many trees at the same time and often leads to overconfidence in the wrong trees. The SH test, though safe to use, may exhibit another type of bias such that it appears conservative. Here I show that the AU test is less biased than other methods in typical cases of tree selection. These points are illustrated in a simulation study as well as in the analysis of mammalian mitochondrial protein sequences. The theoretical argument provides a simple formula that covers the bootstrap probability test, the Kishino-Hasegawa test, the AU test, and the Zharkikh-Li test. A practical suggestion is provided as to which test should be used under particular circumstances. PMID- 12079647 TI - Model misspecification and probabilistic tests of topology: evidence from empirical data sets. AB - Probabilistic tests of topology offer a powerful means of evaluating competing phylogenetic hypotheses. The performance of the nonparametric Shimodaira-Hasegawa (SH) test, the parametric Swofford-Olsen-Waddell-Hillis (SOWH) test, and Bayesian posterior probabilities were explored for five data sets for which all the phylogenetic relationships are known with a very high degree of certainty. These results are consistent with previous simulation studies that have indicated a tendency for the SOWH test to be prone to generating Type 1 errors because of model misspecification coupled with branch length heterogeneity. These results also suggest that the SOWH test may accord overconfidence in the true topology when the null hypothesis is in fact correct. In contrast, the SH test was observed to be much more conservative, even under high substitution rates and branch length heterogeneity. For some of those data sets where the SOWH test proved misleading, the Bayesian posterior probabilities were also misleading. The results of all tests were strongly influenced by the exact substitution model assumptions. Simple models, especially those that assume rate homogeneity among sites, had a higher Type 1 error rate and were more likely to generate misleading posterior probabilities. For some of these data sets, the commonly used substitution models appear to be inadequate for estimating appropriate levels of uncertainty with the SOWH test and Bayesian methods. Reasons for the differences in statistical power between the two maximum likelihood tests are discussed and are contrasted with the Bayesian approach. PMID- 12079648 TI - Type 1 error rates of the parsimony permutation tail probability test. PMID- 12079649 TI - Preferential host switching by primate lentiviruses can account for phylogenetic similarity with the primate phylogeny. AB - Primate lentiviruses (PLV) from closely related primate species have been observed to be more closely related to each other than to PLV from more distantly related primate species. The current explanation for this observation is the codivergence hypothesis; that is, the divergence of a virus lineage results from the divergence of the host lineage. We show that, alternatively, frequent cross species transmission of PLV, coupled with a tendency for more closely related primate species to exchange viruses "successfully," can result in apparent codivergence. This host-switching hypothesis reconciles several puzzling observations related to the evolution of PLV. PMID- 12079651 TI - Facial resemblance enhances trust. AB - Organisms are expected to be sensitive to cues of genetic relatedness when making decisions about social behaviour. Relatedness can be assessed in several ways, one of which is phenotype matching: the assessment of similarity between others' traits and either one's own traits or those of known relatives. One candidate cue of relatedness in humans is facial resemblance. Here, I report the effects of an experimental manipulation of facial resemblance in a two-person sequential trust game. Subjects were shown faces of ostensible playing partners manipulated to resemble either themselves or an unknown person. Resemblance to the subject's own face raised the incidence of trusting a partner, but had no effect on the incidence of selfish betrayals of the partner's trust. Control subjects playing with identical pictures failed to show such an effect. In a second experiment, resemblance of the playing partner to a familiar (famous) person had no effect on either trusting or betrayals of trust. PMID- 12079652 TI - A covarion-based method for detecting molecular adaptation: application to the evolution of primate mitochondrial genomes. AB - A new method for detecting site-specific variation of evolutionary rate (the so called covarion process) from protein sequence data is proposed. It involves comparing the maximum-likelihood estimates of the replacement rate of an amino acid site in distinct subtrees of a large tree. This approach allows detection of covarion at the gene or the amino acid levels. The method is applied to mammalian mitochondrial-protein sequences. Significant covarion-like evolution is found in the (simian) primate lineage: some amino acid positions are fast-evolving (i.e. unconstrained) in non-primate mammals but slow-evolving (i.e. highly constrained) in primates, and some show the opposite pattern. Our results indicate that the mitochondrial genome of primates reached a new peak of the adaptive landscape through positive selection. PMID- 12079653 TI - On the evolution of virulence and the relationship between various measures of mortality. AB - Smallpox causes roughly 20% mortality whereas chickenpox causes less than 0.1%. Most 'verbal' (i.e. non-mathematical) discussions using a mortality definition of virulence would therefore label smallpox as more virulent. Indeed, the virulence of many diseases is measured using such case mortalities, chi, or related measures such as expected host lifespan, T, or lethal dose, LD(x). But chi, T and LD(x) are only indirectly related to parasite-induced instantaneous mortality rate, alpha, which is the mortality measure used in much of the theory developed to explain virulence evolution. Here I point out that relatively deadly pathogens can actually have lower values of alpha than benign pathogens, demonstrating that alpha does not, by itself, reflect the extent to which a parasite causes host mortality. I present mathematical relationships between alpha and chi, T and LD(x), and use these to demonstrate that predictions about virulence evolution can be qualitatively altered depending upon which measure is used as the definition of virulence. Two simple examples are presented to illustrate this point, one of which demonstrates that the well-cited prediction that virulence should evolve to be higher when disease-independent host mortality increases need not hold. This prediction has been made in terms of parasite-induced instantaneous mortality, alpha, but if virulence is measured using case mortality (or T or LD(x)) then this prediction can easily be reversed. Theoretical and empirical researchers must use compatible mortality measures before a productive exchange between the two can take place, and it is suggested that case mortality (or lethal dose) is best suited as a single (mortality) measure of parasite virulence. PMID- 12079654 TI - Male phenotype predicts insemination success in guppies. AB - Theory predicts that mate choice can lead to an increase in female fecundity if the secondary sexual traits used by females to assess male quality covary with the number of sperm transferred during copulation. Where females mate multiply, such a relationship between male attractiveness and ejaculate size may, additionally (or alternatively), serve to augment the effect of indirect selection by biasing paternity in favour of preferred males. In either case, a positive correlation between male attractiveness and the size of ejaculates delivered at copulation is predicted. To date, some of the most convincing (indirect) evidence for this prediction comes from the guppy, a species of fish exhibiting a resource-free mating system in which attractive males tend to have larger sperm reserves. We show that, during solicited copulations, male guppies with preferred phenotypes actually transfer more sperm to females than their less ornamented counterparts, irrespective of the size of their initial sperm stores. Our results also reveal that, during coercive copulations, the relationship between ejaculate size and the male's phenotype breaks down. This latter result, in conjunction with our finding that mating speed--a factor under the female's control-is a significant predictor of ejaculate size, leads us to speculate that females may exert at least partial control over the number of sperm inseminated during cooperative matings. PMID- 12079655 TI - The sexual selection continuum. AB - The evolution of mate choice for genetic benefits has become the tale of two hypotheses: Fisher's 'run-away' and 'good genes', or viability indicators. These hypotheses are often pitted against each other as alternatives, with evidence that attractive males sire more viable offspring interpreted as support for good genes and with a negative or null relationship between mating success of sons and other components of fitness interpreted as favouring the Fisher process. Here, we build a general model of female choice for indirect benefits that captures the essence of both the 'Fisherian' and 'good-genes' models. All versions of our model point to a single process that favours female preference for males siring offspring of high reproductive value. Enhanced mating success and survival are therefore equally valid genetic benefits of mate choice, but their relative importance varies depending on female choice costs. The relationship between male attractiveness and survival may be positive or negative, depending on life history trade-offs and mating skew. This relationship can change sign in response to increased costliness of choice or environmental change. Any form of female preference is subject to self-reinforcing evolution, and any relationship (or lack thereof) between male display and offspring survival is inevitably an indicator of offspring reproductive values. Costly female choice can be maintained with or without higher offspring survival. PMID- 12079656 TI - Sexual selection constrained by life history in a butterfly. AB - Trait evolution via sexual selection has traditionally been viewed as isolated from life-history constraints. Recent theoretical treatments, however, predict that costly sexually selected characters should be mediated by the same allocational trade-offs that apply to more conventional aspects of reproductive investment. Participation in risky competitive behaviours, for example male-male combat, should therefore increase as the opportunity for future reproduction declines. However, the demonstration of such trade-offs has proven to be elusive due to concomitant age-based variation in the physical determinants of fighting 'ability'. Here, I exploit the unique nature of a butterfly contest system to provide compelling evidence for lifetime partitioning of risky and aggressive sexually selected behaviours. I show that male Hypolimnas bolina become more willing to persist in contests over mating territories, and more generally accepting of injury risks, as they age. Contest persistence in this species is not mediated simply by physical condition, and I experimentally isolate the effect of ageing per se from resource ownership and previous contest experience. These results demonstrate how sexually selected behaviours can be ultimately mediated by a shifting trade-off between contemporary reproductive effort and future opportunities. PMID- 12079657 TI - Bower decorations attract females but provoke other male spotted bowerbirds: bower owners resolve this trade-off. AB - Elaborate secondary sexual traits offset the costs that they impose on their bearer by facilitating reproductive benefits, through increased success in intrasexual contests or increased attractiveness to choosy mates. Some traits enhance both strategies. Conversely, I show that spotted bowerbirds Chlamydera maculata may face a trade-off. The trait that best predicts their mating success, numbers of Solanum berries exhibited on a bower, also provokes increased intrasexual aggression in the form of bower destructions by neighbouring bower owners, which reduce the quality of the male's bower. At natural berry numbers, levels of mating success in the population are skewed, but levels of destruction do not vary with berry number. When berry numbers are artificially exaggerated, increased levels of destructions occur, but mating success does not increase. When offered excess berries, either to add to the bower or artificially placed on the bower, bower owners preferred to use numbers of berries related to the number that they displayed naturally. This decision is made without direct experience of the attendant changes in destruction or mating success. This indicates that bower owners may assess their own social standing in relation to their neighbours and modulate their display accordingly. PMID- 12079658 TI - Sound improves visual discrimination learning in avian predators. AB - Aposematic insects use warning colours to deter predators, but many also produce odours or sounds when attacked by a predator. One possible role for these additional components is that they promote the association between the warning colour and the non-profitability it signals, thus reducing the chance of future attacks from visually hunting predators. This experiment explicitly tests this idea by looking at the effects of sound on a visual discrimination task. Young domestic chicks were trained to look for food rewards under coloured paper cones scattered in an experimental arena. In a subsequent visual discrimination task, they learned to discriminate between rewarded and non-rewarded hats on the basis of colour. Half the chicks performed this task in silence, whilst the other half had a tone played when they attacked non-rewarded hats. The presence of the tone improved the speed of colour discrimination learning. This demonstrates that there could be a selective advantage for aposematic coloured insects to emit sounds when attacked, since avian predators will learn to avoid their coloration more quickly. The role of psychological interactions between signal components in receivers is discussed in relation to the evolution of multimodal displays. PMID- 12079659 TI - The 'island rule' in birds: medium body size and its ecological explanation. AB - Do birds show a different pattern of insular evolution from mammals? Mammals follow the 'island rule', with large-bodied species getting smaller on islands and small-bodied species getting bigger. By contrast, the traditional view on birds is that they follow no general island rule for body size, but that there is an insular trend for large bills. Insular shifts in feeding ecology are, therefore, widely assumed to be the primary cause of divergence in island birds. We use a comparative approach to test these ideas. Contrary to the traditional view, we find no evidence for increased bill size in insular populations. Instead, changes in both bill size and body size obey the 'island rule'. The differences between our results and the traditional view arise because previous analyses were based largely on passerines. We also investigate some ecological factors that are thought to influence island evolution. As predicted by the traditional view, shifts in bill size are associated with feeding ecology. By contrast, shifts in body size are associated with the potential for intraspecific competition and thermal ecology. All these results remain qualitatively unchanged when we use different methods to score the ecological factors and restrict our analyses to taxa showing pronounced morphological divergence. Because of strong covariation between ecological factors, however, we cannot estimate the relative importance of each ecological factor. Overall, our results show that the island rule is valid for both body size and bill length in birds and that, in addition to feeding ecology, insular shifts in the level of intraspecific competition and the abiotic environment also have a role. PMID- 12079660 TI - Spleen volume varies with colony size and parasite load in a colonial bird. AB - Comparisons across bird species have indicated that those more exposed to parasites and pathogens invest more in immunological defence, as measured by spleen size. We investigated how spleen volume varied with colony size, parasite load and an individual's colony-size history in the cliff swallow, Petrochelidon pyrrhonota, a colonial passerine bird of North America. We used a sample of over 1700 birds that had all died during a period of inclement weather in 1996. We experimentally manipulated ectoparasitism by fumigating nests in some colonies prior to the bad weather. Birds from parasite-free colonies had significantly smaller spleens than those from naturally infested sites; spleen volume did not differ between the sexes and did not vary with age. Mean spleen volume increased significantly with the colony size at a site prior to the bad weather in 1996 and at the site in 1995, both measures of colony size being indices of ectoparasitism at a site. An individual's history of breeding-colony size (defined as the average colony size it had occupied in years prior to 1996) had no association with its spleen size. The results are consistent with parasite-induced splenomegaly whenever birds are exposed to large numbers of ectoparasites. The results do not support spleen size as being a signal of differential life-history investment in immunological defence among individuals and thus run counter to interpretations from recent cross-species comparisons. PMID- 12079661 TI - Not as the crow flies: a historical explanation for circuitous migration in Swainson's thrush (Catharus ustulatus). AB - Many migratory songbirds follow circuitous migratory routes instead of taking the shortest path between overwintering and breeding areas. Here, we study the migration patterns in Swainson's thrush (Catharus ustulatus), a neartic neotropical migrant songbird, using molecular genetic approaches. This species is presently separated into genetically distinct coastal and continental populations that diverged during the Late Pleistocene (as indicated by molecular dating), yet appear to have retained ancestral patterns of migration. Low nucleotide diversity, a star-like haplotype phylogeny and unimodal mismatch distributions all support the hypothesis that both the coastal and the continental populations have undergone recent demographic expansions. Nearctic-neotropical banding and genetic data show nearly complete segregation of migratory routes and of overwintering locations: coastal populations migrate along the Pacific Coast to overwintering sites in Central America and Mexico, whereas continental populations migrate along an eastern route to overwintering sites in Panama and South America. Nearctic-neotropical banding data also show that continental birds north, northwest and east of this migratory divide fly thousands of miles east before turning south. We conclude that circuitous migration in the Swainson's thrush is an artefact of a Late Pleistocene range expansion. PMID- 12079662 TI - Deciding on a new home: how do honeybees agree? AB - A swarm of honeybees (Apis mellifera) is capable of selecting one nest-site when faced with a choice of several. We adapt classical mathematical models of disease, information and competing beliefs to such decision-making processes. We show that the collective decision may be arrived at without the necessity for any bee to make any comparison between sites. PMID- 12079663 TI - The potential for floral mimicry in rewardless orchids: an experimental study. AB - More than one-third of orchid species do not provide their pollinators with either pollen or nectar rewards. Floral mimicry could explain the maintenance of these rewardless orchid species, but most rewardless orchids do not appear to have a rewarding plant that they mimic specifically. We tested the hypothesis that floral mimicry can occur through similarity based on corolla colour alone, using naive bumble-bees foraging on arrays of plants with one rewarding model species, and one rewardless putative mimic species (Dactylorhiza sambucina) which had two colour morphs. We found that when bees were inexperienced, they visited both rewardless morphs randomly. However, after bees had gained experience with the rewarding model, and it was removed from the experiment, bees resampled preferentially the rewardless morph most similar to it in corolla colour. This is the first clear evidence, to our knowledge, that pollinators could select for floral mimicry. We suggest that floral mimicry can be a selective force acting on rewardless orchids, but only under some ecological conditions. In particular, we argue that selection on early-flowering rewardless orchids that receive visits from a large pool of naive pollinators will be weakly influenced by mimicry. PMID- 12079664 TI - The persistence of vertically transmitted fungi in grass metapopulations. AB - Theory predicts that (i) vertical transmission of parasites (i.e. when they are passed directly from a host to its offspring) selects for benign association with the host and that (ii) vertically transmitted parasites that lower their hosts' fitness cannot persist if they are not able to infect horizontally (i.e. contagiously) other host individuals in the population. In this paper, we develop a mathematical model to examine whether mutualism is a prerequisite for persistence of exclusively vertically transmitted (from maternal plant to offspring via seeds) fungal endophytes in structured grass metapopulations. Interestingly, endophyte survival does not require plant mutualism, even in a metapopulation consisting of qualitatively identical patches, if vertical transmission of the fungus is perfect, i.e. if all established seedlings in offspring of the endophyte-infected plant are infected. In more realistic situations, when the metapopulation consists of qualitatively different patches, endophyte-infected plants may persist at the metapopulation level even if the vertical transmission is imperfect (due to hyphae inviability or failure to grow into all seeds) and the endophyte decreases the host grass fitness in certain environments. These results have biological importance because they (i) question the requirement of a mutualistic nature in exclusively vertically transmitted symbionts and (ii) emphasize the importance of habitat diversity in relation to symbiont success in vertical transmission. PMID- 12079665 TI - Benefits and costs of mutualism: demographic consequences in a pollinating seed consumer interaction. AB - Interspecific interactions can affect population dynamics and the evolution of species traits by altering demographic rates such as reproduction and survival. The influence of mutualism on population processes is thought to depend on both the benefits and costs of the interaction. However, few studies have explicitly quantified both benefits and costs in terms of demographic rates; furthermore there has been little consideration as to how benefits and costs depend on the demographic effects of factors extrinsic to the interaction. I studied how benefits (pollination) and costs (larval fruit consumption) of pollinating seed consumers (senita moths) affect the reproduction of senita cacti and how these effects may rely on extrinsic water limitation for reproduction. Fruit initiation was not limited by moth pollination, but survival of initiated fruit increased when moth eggs were removed from flowers. Watered cacti produced more flowers and initiated more fruit from hand-pollinated flowers than did unwatered cacti, but fruit initiation remained low despite excess pollen. Even though water, pollination and larvae each affected a component of cactus reproduction, when all of these factors were included in a factorial experiment, pollination and water determined rates of reproduction. Counter-intuitively, larval fruit consumption had a negligible effect on cactus reproduction. By quantifying both benefits and costs of mutualism in terms of demographic rates, this study demonstrates that benefits and costs can be differentially influential to population processes and that interpretation of their influences can depend on demographic effects of factors extrinsic to the interaction. PMID- 12079666 TI - High Wolbachia density in insecticide-resistant mosquitoes. AB - Wolbachia symbionts are responsible for various alterations in host reproduction. The effects of the host genome on endosymbiont levels have often been suggested, but rarely described. Here, we show that Wolbachia density is strongly modified by the presence of insecticide-resistant genes in the common house mosquito, Culex pipiens. The Wolbachia density was estimated using a real-time quantitative PCR assay. Strains harbouring different genes conferring resistance were more infected than a susceptible strain with the same genetic background. We show that this interaction also operates in natural populations. We propose that mosquitoes may control Wolbachia density less efficiently when they carry an insecticide resistant gene, i.e. when they suffer from a physiological resistance cost. PMID- 12079668 TI - Genomics and plant cells: application of genomics strategies to Arabidopsis cell biology. AB - In this review I seek to describe how the complete catalogue of plant genes and proteins, revealed by genome sequencing, can provide novel insights into cell biology. Many new analytical methods have been developed to digest the flood of genome sequence data, including analysis of the transcriptome, proteome and metabolites. High-throughput analysis of protein targeting and other methods will ascribe new information to proteins and create important links with other large datasets. To fulfil the potential revealed by this genomic information, many challenges have to be met. Among these are organizational changes needed to create common datasets accessible to all scientists, and bioinformatics solutions to capture and integrate diverse datasets. Once harnessed, these new strategies will irrevocably change the way we conduct plant science. PMID- 12079669 TI - The shoot apical meristem: the dynamics of a stable structure. AB - The shoot apical meristem (SAM) is a group of proliferating, embryonic-type cells that generates the aerial parts of the plant. SAMs are highly organized and stable structures that can function for years or even centuries. This is in apparent contradiction to the behaviour of their constituent cells, which continuously proliferate and differentiate. To reconcile the dynamic nature of the cells with the stability of the overall system the existence of elaborate signalling networks has been proposed. This is supported by recent work suggesting that the exchange of signals between cells, rather than a rigidly predetermined genetic program, is required for the establishment and functioning of an organized meristem. Together these interactions form a stable network, set up during embryogenesis, that assures the coordination of cell behaviour throughout development. Besides meristem-specific signalling cascades such as the CLAVATA receptor kinase pathway, which controls meristem size, these interactions involve plant hormones. In particular, cytokinins and auxins are implicated in the maintenance of meristem identity and phyllotaxis, respectively. PMID- 12079671 TI - Roles for kinesin and myosin during cytokinesis. AB - Cytokinesis in higher plants involves the phragmoplast, a complex cytoplasmic structure that consists of microtubules (MTs), microfilaments (MFs) and membrane elements. Both MTs and MFs are essential for cell plate formation, although it is not clear which motor proteins are involved. Some candidate processes for motor proteins include transport of Golgi vesicles to the plane of the cell plate and the spatiotemporal organization of the cytoskeletal elements in order to achieve proper deposition and alignment of the cell plate. We have focused on the kinesin like calmodulin binding protein (KCBP) and, more broadly, on myosins. Using an antibody that constitutively activates KCBP, we find that this MT motor, which is minus-end directed, contributes to the organization of the spindle and phragmoplast MTs. It does not participate in vesicle transport; rather, because of the orientation of the phragmoplast MTs, it is supposed that plus-end kinesins fill this role. Myosins, on the other hand, based on their inhibition with 2,3 butanedione monoxime and 1-(5-iodonaphthalene-1-sulphonyl)-1H-hexahydro-1,4 diazepine (ML-7), are associated with the process of post-mitotic spindle/phragmoplast alignment and with late lateral expansion of the cell plate. They are also not the principal motors involved in vesicle transport. PMID- 12079670 TI - Plant D-type cyclins and the control of G1 progression. AB - The basic pattern of controls that operate during the G1 phase of the plant cell cycle shows much closer similarity to animals than to the yeasts and other fungi. The activity of D-type cyclin (CycD) kinases is induced in response to stimulatory signals, and these phosphorylate the plant homologue of the retinoblastoma tumour susceptibility (Rb) protein. It is likely that Rb phosphorylation results in the activation of genes under the control of E2F transcription factors, including those required for S phase entry. As the initial triggers of the cascade, attention has focused on the CycDs, and a family of 10 genes is present in Arabidopsis, divided into three major and three minor groups. Analysis to date suggests that these groups are functionally distinct. PMID- 12079672 TI - Control of plant cytokinesis by an NPK1-mediated mitogen-activated protein kinase cascade. AB - Cytokinesis is the last essential step in the distribution of genetic information to daughter cells and partition of the cytoplasm. In plant cells, various proteins have been found in the phragmoplast, which corresponds to the cytokinetic apparatus, and in the cell plate, which corresponds to a new cross wall, but our understanding of the functions of these proteins in cytokinesis remains incomplete. Reverse genetic analysis of NPK1 MAPKKK (nucleus- and phragmoplast-localized protein kinase 1 mitogen-activated protein kinase kinase kinase) and investigations of factors that might be functionally related to NPK1 have helped to clarify new aspects of the mechanisms of cytokinesis in plant cells. In this review, we summarize the evidence for the involvement of NPK1 in cytokinesis. We also describe the characteristics of a kinesin-like protein and the homologue of a mitogen-activated protein kinase that we identified recently, and we discuss possible relationships among these proteins in cytokinesis. PMID- 12079675 TI - Molecular genetic analysis of left-right handedness in plants. AB - Handedness in plant growth may be most familiar to us when we think of tendrils or twining plants, which generally form consistent right- or left-handed helices as they climb. The petals of several species are sometimes arranged like fan blades that twist in the same direction. Another less conspicuous example is 'circumnutation', the oscillating growth of axial organs, which alternates between a clockwise and an anti-clockwise direction. To unravel molecular components and cellular determinants of handedness, we screened Arabidopsis thaliana seedlings for helical growth mutants with fixed handedness. Recessive spiral1 and spiral2 mutants show right-handed helical growth in roots, hypocotyls, petioles and petals; semi-dominant lefty1 and lefty2 mutants show opposite left-handed growth in these organs. lefty mutations are epistatic to spiral mutations. Arabidopsis helical growth mutants with fixed handedness may be impaired in certain aspects of cortical microtubule functions, and characterization of the mutated genes should lead us to a better understanding of how microtubules function in left-right handedness in plants. PMID- 12079674 TI - Actin-binding proteins in the Arabidopsis genome database: properties of functionally distinct plant actin-depolymerizing factors/cofilins. AB - The plant actin cytoskeleton is a highly dynamic, fibrous structure essential in many cellular processes including cell division and cytoplasmic streaming. This structure is stimulus responsive, being affected by internal stimuli, by biotic and abiotic stresses mediated in signal transduction pathways by actin-binding proteins. The completion of the Arabidopsis genome sequence has allowed a comparative identification of many actin-binding proteins. However, not all are conserved in plants, which possibly reflects the differences in the processes involved in morphogenesis between plant and other cells. Here we have searched for the Arabidopsis equivalents of 67 animal/fungal actin-binding proteins and show that 36 are not conserved in plants. One protein that is conserved across phylogeny is actin-depolymerizing factor or cofilin and we describe our work on the activity of vegetative tissue and pollen-specific isoforms of this protein in plant cells, concluding that they are functionally distinct. PMID- 12079673 TI - Cytoskeleton and plant organogenesis. AB - The functions of microtubules and actin filaments during various processes that are essential for the growth, reproduction and survival of single plant cells have been well characterized. A large number of plant structural cytoskeletal or cytoskeleton-associated proteins, as well as genes encoding such proteins, have been identified. Although many of these genes and proteins have been partially characterized with respect to their functions, a coherent picture of how they interact to execute cytoskeletal functions in plant cells has yet to emerge. Cytoskeleton-controlled cellular processes are expected to play crucial roles during plant cell differentiation and organogenesis, but what exactly these roles are has only been investigated in a limited number of studies in the whole plant context. The intent of this review is to discuss the results of these studies in the light of what is known about the cellular functions of the plant cytoskeleton, and about the proteins and genes that are required for them. Directions are outlined for future work to advance our understanding of how the cytoskeleton contributes to plant organogenesis and development. PMID- 12079676 TI - The mechanics of cell fate determination in petals. AB - The epidermal cells of petals of many species are specialized, having a pronounced conical shape. A transcription factor, MIXTA, is required for the formation of conical cells in Antirrhinum majus; in shoot epidermal cells of several species, expression of this gene is necessary and sufficient to promote conical cell formation. Ectopic expression has also shown MIXTA to be able to promote the formation of multicellular trichomes, indicating that conical cells and multicellular trichomes share elements of a common developmental pathway. Formation of conical cells or trichomes is also mutually exclusive with stomatal formation. In Antirrhinum, MIXTA normally only promotes conical cell formation on the inner epidermal layer of the petals. Its restricted action in cell fate determination results from its specific expression pattern. Expression of MIXTA, in turn, requires the activity of B-function genes, and biochemical evidence suggests that the products of DEFICIENS, GLOBOSA and SEPALLATA-related genes directly activate MIXTA expression late in petal development, after the completion of cell division in the petal epidermis. A MIXTA-like gene, AmMYBML1, is also expressed in petals. AmMYBML1 expression is high early in petal development. This gene may direct the formation of trichomes in petals. In specifying the fates of different cell types in petals, regulatory genes like MIXTA may have been duplicated. Changes in the timing and spatial localization of expression then provides similar regulatory genes which specify different cell fates. PMID- 12079677 TI - Building a hair: tip growth in Arabidopsis thaliana root hairs. AB - The Arabidopsis thaliana root hair is used as a model for studying tip growth in plants. We review recent advances, made using physiological and genetic approaches, which give rise to different, yet compatible, current views of the establishment and maintenance of tip growth in epidermal cells. For example, an active calcium influx channel localized at the tip of Arabidopsis root hairs has been identified by patch-clamp measurements. Actin has been visualized in vivo in Arabidopsis root hairs by using a green-fluorescent-protein-talin reporter and shown to form a dense mesh in the apex of the growing tip. The kojak gene, which encodes a protein similar to the catalytic subunit of cellulose synthase, is needed in the first stages of hair growth. A role for LRX1, a leucine-rich repeat extensin, in determining the morphology of the cell wall of root hairs has been established using reverse genetics. The new information can be integrated into a general and more advanced view of how these specialized plant cells grow. PMID- 12079679 TI - Pharmacological properties of rat alpha 7 nicotinic receptors expressed in native and recombinant cell systems. AB - The pharmacological properties of the rat alpha7 nicotinic acetylcholine receptor endogenously expressed in PC12 cells and recombinantly expressed in GH4C1 cells (alpha7-GH4C1 cells) were characterized and compared. Patch-clamp recordings demonstrated that activation by choline and block by methyllycaconitine and dihydro-beta-erythroidine were similar, but block by mecamylamine was different. Whereas in alpha7-GH4C1 cells the inhibition curve for mecamylamine was monophasic (IC(50) of 1.6 microM), it was biphasic in PC12 cells (IC(50) values of 341 nM and 9.6 microM). The same rank order of potency was obtained for various nicotinic agonists, while acetylcholine was 3.7-fold less potent and 1.5 fold more effective in PC12 cells. Dihydro-beta-erythroidine differentially blocked acetylcholine-evoked currents in both systems. Since reverse transcriptase polymerase chain reaction (RT-PCR) experiments revealed expression of alpha3, alpha4, alpha5, alpha7 and beta4 subunits in PC12 cells, whereas GH4C1 cells express only the beta4 subunit, our results suggest that more than one form of alpha7 containing heteromeric nicotinic receptors might be functionally expressed in PC12 cells. PMID- 12079678 TI - Trichome morphogenesis: a cell-cycle perspective. AB - Arabidopsis leaf hairs (trichomes) are polyploid epidermal cells with a predictable branching pattern. More than 15 genes have been identified that are involved in the regulation of branching. The cloning of the ZWICHEL, ANGUSTIFOLIA and STICHEL genes points to two mechanistic aspects of branch formation: (i) a role of the microtubule cytoskeleton; and (ii) a link to the regulation of cell divisions. The latter aspect is supported by the recent identification of an Arabidopsis mutant with multicellular trichomes, the siamese mutant, suggesting that Arabidopsis trichomes are evolutionarily derived from multicellular forms. We speculate that the spatial information for branch formation is derived from mechanisms employed in cell divisions. PMID- 12079680 TI - Ethanol differentially modulates the expression and activity of cell cycle regulatory proteins in rat aortic smooth muscle cells. AB - The aim of this study was to determine the effect of ethanol on cell cycle events during the G(1) and S phases in cultured vascular smooth muscle cells (VSMC). Flow cytometric analysis for the DNA content in rat aortic VSMC indicated that following ethanol treatment, the cell population in the G(0)/G(1) phase increased; 57.8+/-1.6% vs. 72.3+/-1.2%, concomitant with a decrease in cells in the S phase; 12.7+/-1.4% vs. 3.67+/-0.6%, for control vs. ethanol, respectively. Western blot analysis on VSMC lysates demonstrated that ethanol (10-160 mmol/l) dose-dependently inhibited serum-induced retinoblastoma (pRb) hyperphosphorylation. While having no effect on Cdk2 protein expression, ethanol dose-dependently decreased (IC(50) approximately 60 mmol/l) Cdk2 activity, assessed by histone H1 phosphorylation. Furthermore, ethanol induced the expression of the cyclin-dependent kinase (Cdk) inhibitor p21(waf1/cip1), and inhibited the induction of cyclin A. These data demonstrate that modulation of the expression and activity of key cell cycle regulatory molecules may be a mechanism by which ethanol inhibits VSMC proliferation. These actions of ethanol may be relevant to its cardiovascular protective effect in vivo. PMID- 12079681 TI - Effects of streptomycin sulphate on I(CaL), I(Kr) and I(Ks) in guinea-pig ventricular myocytes. AB - In single guinea pig ventricular myocytes, streptomycin sulphate (streptomycin) reduced intracellular Ca(2+) transients (IC(50) 1.9 mM) and contractility (IC(50) 1.0 mM), 2 mM streptomycin prolonged the action potential. Under switch voltage clamp, 2 mM streptomycin reduced the L-type Ca(2+) current (I(CaL)) amplitude and (Ca(2+)-dependent) relative inactivation at positive membrane potentials and reduced the rapid and slow components of the delayed rectifier current (I(K)). This latter effect seemed Ca(2+)-dependent, not being seen when nifedipine and BAPTA were used to reduce intracellular Ca(2+). Fifty micromolars of streptomycin had no significant effects on any parameter studied. We conclude that the negative inotropic effect of streptomycin results from blockade of I(CaL), and thus, a reduction of intracellular Ca(2+) transients, while prolongation of the action potential is more consistent with effects on I(K). These observations link mechanical and electrical effects of streptomycin that may be important, for example, when streptomycin is used to block stretch-activated events in cardiac muscle. PMID- 12079682 TI - The antinociceptive effect of tramadol in the formalin test is mediated by the serotonergic component. AB - The aim of this study was to investigate the neurotransmissions involved in the antinociceptive effect of tramadol in the formalin test, which is an animal model of acute and tonic pain. A subcutaneous injection of formalin produces a biphasic nociceptive response: phase 1 (0-10 min-acute pain) and phase 2 (21-60 min-tonic pain). Nociceptive activity is reduced greatly during the 10 min between these two phases. We measured in mice the effects of (+/-)-tramadol, and of (+)- and ( )-tramadol administered before the induction of pain by formalin, in the presence and absence of drugs that act on the opioidergic, serotonergic and noradrenergic systems (naloxone, ketanserin, fluoxetine, maprotiline). With respect to animals treated with formalin alone, (+/-)-tramadol and its enantiomers significantly reduced the duration of nociceptive behaviours (lifting, licking, favouring, shaking, and flinching of the formalin-treated paw) during phase 2. This effect was prevented by the 5-HT(2) receptor antagonist ketanserin, but not by naloxone which, on the contrary, was able to prevent the antinociceptive effect of morphine. Naloxone and ketanserin did not affect the duration of nociceptive behaviour in animals not treated with tramadol. Fluoxetine (a selective 5 hydroxytryptamine (5-HT) reuptake inhibitor), but not maprotiline (a selective norepinephrine reuptake inhibitor), potentiated the antinociceptive effect of (+/ )-tramadol. In conclusion, we demonstrate that the serotonergic pathway is responsible for the antinociceptive effect of tramadol in phase 2 of the formalin test, and that this effect is mediated by 5-HT(2) receptors. PMID- 12079683 TI - Involvement of imidazoline receptors in the centrally acting muscle-relaxant effects of tizanidine. AB - The centrally acting muscle relaxant tizanidine has an imidazoline structure and binds not only to alpha(2)-adrenoceptors but also to imidazoline receptors. The role of imidazoline receptors in the muscle-relaxant effect of tizanidine was studied using the alpha(2)-adrenoceptor/imidazoline receptor antagonist idazoxan and the alpha(2)-adrenoceptor antagonist yohimbine. Tizanidine decreased the spinal mono- and polysynaptic reflexes in intact rats, and the inhibitory effects were antagonized by idazoxan but not by yohimbine. After pretreatment with prazosin, tizanidine decreased the mono- and polysynaptic reflexes in spinalized rats. While yohimbine partly inhibited tizanidine-induced depression of the polysynaptic reflex, idazoxan completely abolished tizanidine-induced depression of spinal reflexes. Furthermore, tizanidine-induced muscle relaxation in the traction test was significantly inhibited by idazoxan but not by yohimbine. From these results, it is suggested that imidazoline receptors, but not alpha(2) adrenoceptors, are involved in the supraspinal inhibitory effects of tizanidine on spinal reflexes, and at the spinal level, alpha(2)-adrenoceptors and imidazoline receptors are involved in the inhibitory effects of tizanidine. PMID- 12079684 TI - Proconvulsive effect of the GABA(B) receptor antagonist, SCH 50911, in rats undergoing ethanol withdrawal syndrome. AB - The present study investigated the effect of the GABA(B) receptor antagonist, SCH 50911 [(2S)(+)-5,5-dimethyl-2-morpholineacetic acid], on the occurrence of seizures in ethanol-dependent rats undergoing ethanol withdrawal syndrome. The acute administration of nonconvulsive doses of SCH 50911 (0, 100, 170 and 300 mg/kg, i.p.) resulted in a dramatic facilitation of spontaneous seizure occurrence. This finding, together with the reported ability of the GABA(B) receptor agonist, baclofen, to suppress seizures associated to ethanol withdrawal syndrome, suggests that the GABA(B) receptor may be part of the neural substrate underlying the hyperexcitability of ethanol withdrawal syndrome. PMID- 12079685 TI - Depressant effect on a C-fibre reflex in the rat, of RB101, a dual inhibitor of enkephalin-degrading enzymes. AB - The effect of N-[(R,S)-2-benzyl-3[(S)-(2-amino-4-methylthio)butyldithiol]-1 oxopropyl]-L-phenylalanine benzyl ester (RB101), a dual inhibitor of the enkephalin-degrading enzymes, neutral endopeptidase and aminopeptidase N, was assessed in anaesthetised rats on the C-fibre reflex elicited by electrical stimulation within the sural nerve territory and recorded from the ipsilateral biceps femoris muscle. The temporal evolution of the pharmacological response was monitored by the repeated application of a constant stimulus intensity, namely three times threshold (3 T). In addition, recruitment curves were built by varying the stimulus intensity from 0 to 7 T. RB101 (7.5, 15 and 30 mg kg(-1), i.v.) induced a dose-dependent, naloxone-reversible depression of the reflex, which lasted around 60 min with the highest dose. The ED(50) was calculated as 16.9 mg kg(-1). Analyses of the recruitment curves revealed: (1) a significant increase of threshold; (2) a significant depression of the reflex in the ascending part of the curve; and (3) a lack of major depressive effects on the responses elicited by the strongest stimuli (corresponding to the plateau of the curve). The increase in the nociceptive threshold by enkephalin-degrading enzyme inhibitors, confirms previous data obtained from behavioural tests. In addition, the present study revealed an efficacy of these compounds over a wide range of stimulus intensities, albeit excluding the highest. PMID- 12079686 TI - Low molecular weight glycosaminoglycan blockade of beta-amyloid induced neuropathology. AB - Previous studies have shown different roles for proteoglycans and glycosaminoglycans (GAGs) in Alzheimer's disease (AD) neuropathology. Using a rat model of beta-amyloid induced neuropathology, we tested whether low molecular weight glycosaminoglycans (Certoparin and C6) could be useful as preventative agents and/or as a potential therapeutic treatment for AD. Chronic subcutaneous low molecular weight glycosaminoglycan injections beginning either before or after an intra-amygdaloid beta-amyloid-(25-35) injection blocked abnormal intracellular tau changes and reactive astrocytosis but did not affect beta amyloid's aggregation state. Also, low molecular weight glycosaminoglycan injections beginning 1 day prior to sacrifice did not block the effects of beta amyloid nor did injections of a disaccharide, suggesting chronic low molecular weight glycosaminoglycan treatment is needed to block the effects of beta amyloid. Furthermore, these data indicate that there is a molecular weight range of active low molecular weight glycosaminoglycans in this model; and supports the investigation of low molecular weight glycosaminoglycans as a preventative and/or therapeutic treatment of beta-amyloid induced neuropathology. PMID- 12079687 TI - Involvement of 5-hydroxytryptamine neuronal system in Delta(9) tetrahydrocannabinol-induced impairment of spatial memory. AB - The present study investigated the involvement of the serotonin (5 hydroxytryptamine, 5-HT) neuronal system in the Delta(9)-tetrahydrocannabinol induced impairment of spatial memory in the eight-arm radial maze in rats. Delta(9)-Tetrahydrocannabinol (6 mg/kg, i.p.), which impairs spatial memory, significantly increased the 5-HT content in the ventral hippocampus. A microdialysis study showed that Delta(9)-tetrahydrocannabinol (6 mg/kg, i.p.) decreased 5-HT release in the ventral hippocampus. The 5-HT precursor, 5-hydroxy L-tryptophan (5-HTP; 50 mg/kg, i.p.), the 5-HT re-uptake inhibitor, clomipramine (0.01 and 0.1 mg/kg, i.p.), the 5-HT receptor agonist, 5-methoxy-N,N dimethyltryptamine (5-MeODMT; 0.01 and 0.03 mg/kg, i.p.), and the 5-HT(2) receptor agonist, 1-(2,5-dimethoxy 4-iodophenyl)-2-amino propane (DOI; 10 microg/kg, i.p.), significantly attenuated the Delta(9)-tetrahydrocannabinol induced impairment of spatial memory. These results suggest that the 5-HT neuronal system may be involved in the Delta(9)-tetrahydrocannabinol-induced impairment of spatial memory. PMID- 12079688 TI - A comparison of the anti-nociceptive effects of voltage-activated Na+ channel blockers in the formalin test. AB - We have used the rat formalin test to compare the anti-nociceptive properties of several voltage-activated Na(+) channel blockers. The antiarrhthymic mexiletine (37.5 and 50 mg/kg, i.p.) attenuated flinching behaviour in both first and second phases of the test compared with vehicle (P<0.05). The anti-convulsants lamotrigine (15 and 30 mg/kg, i.p.) and carbamazepine (20 mg/kg, i.p.) also inhibited second phase flinching behaviour compared with vehicle (P<0.05), although phenytoin (up to 40 mg/kg, i.p.) was without effect. Riluzole (5 mg/kg, i.p.), in contrast to lubeluzole (up to 10 mg/kg, i.p.) also inhibited second phase flinching behaviour compared with vehicle (P<0.05). When tested against an acute thermal nociceptive stimulus mexiletine, lubeluzole and riluzole exhibited anti-nociceptive effects. The anti-nociceptive doses used in the formalin test produced no motor impairment in the rotarod test. Thus, voltage-activated Na(+) channel blockers can attenuate nociceptive behaviour in the formalin test, and a specific mechanism of action on Na(+) channel function may be required for this to occur. PMID- 12079689 TI - Ca2+ channel blocking effect of iso-S-petasin in rat aortic smooth muscle cells. AB - The purpose of the present study was to examine the mechanisms underlying the putative hypotensive actions of iso-S-petasin, a sesquiterpene extract of Petasites formosanus through both in vivo and in vitro experiments. Intravenous administration of iso-S-petasin elicited dose-dependent (0.1-1.5 mg/kg) hypotensive and bradycardiac responses in anesthetized rats. Isometric tension recording in isolated thoracic aorta revealed that iso-S-petasin (0.01-100 microM) inhibited KCl- or Bay K 8644 (1,4-dihydro-2,6-dimethyl-5-nitro-4-[2' (trifluoromethyl)phenyl]-3-pyridinecarboxylic acid methyl ester)-induced vasoconstriction independent of endothelium. Iso-S-Petasin also attenuated Ca(2+) induced vasoconstriction in a concentration-dependent manner in Ca(2+) depleted/high K(+)-depolarized ring segments, indicating that iso-S-petasin inhibited Ca(2+) influx into vascular smooth muscle cells. This was confirmed by whole-cell patch-clamp recording in cultured vascular smooth muscle cells where iso-S-petasin (10-100 microM) appeared to directly inhibit the L-type voltage dependent Ca(2+) channel (VDCC) activity. Intracellular Ca(2+) concentration ([Ca(2+)](i)) measurements using the fluorescent probe fura-2/AM (1-[2-(5 carboxyoxazol-2-yl)-6-aminobenzofuran-5-oxy]-2-(2'-amino-5'-methylphenoxy)-ethane N,N,N',N'-tetraacetic acid pentaacetoxymethyl ester) showed suppression of the KCl-stimulated increase in [Ca(2+)](i) by iso-S-petasin (10, 100 microM). In conclusion, these results suggest that Ca(2+) antagonism of the L-type VDCC in vascular smooth muscle cells might largely account for the hypotensive action of iso-S-petasin. PMID- 12079691 TI - Triphenylethylene antiestrogen-induced acute relaxation of mouse duodenal muscle. Possible involvement of Ca2+ channels. AB - The nonsteroidal antiestrogens tamoxifen, 4-OH-tamoxifen and toremifene rapidly inhibited spontaneous contractile activity and reduced basal tone in isolated mouse duodenal muscle. Inhibition was rapid in onset ( approximately 2 min) and was not mimicked by the pure steroidal antiestrogen 7alpha-[9-[(4,4,5,5,5 pentafluoropentyl)sulfinyl]nonyl]-estra-1,3,5(10)-triene-3,17beta-diol (ICI182,780) indicating the involvement of non-genomic mechanisms. Inhibition by tamoxifen and 4-OH-tamoxifen were observed at concentrations comparable to those reached in antiestrogen adjuvant therapy. Antiestrogen-relaxed tissues showed no response to KCl depolarisation or K(+) channel blockade but displayed clear transient responses to acethylcholine or to the muscarinic receptor agonist carbachol. Frequency analysis showed that spontaneous activity could be readily restored in antiestrogen-relaxed tissues by the exposure to the L-type Ca(2+) channel agonist 1,4-dihydro-2,6-dimethyl-5-nitro-4-[2-(trifluoromethyl)phenyl] pyridine-3-carboxilic acid methyl ester (BAY K8644). Our experiments suggest that triphenylethylene antiestrogens relax duodenal intestinal muscle via a mechanism that involves inhibition of L-type Ca(2+) channels but not activation of K(+) channels. PMID- 12079690 TI - Involvement of p44/42 mitogen-activated protein kinases in regulating angiotensin II- and endothelin-1-induced contraction of rat thoracic aorta. AB - In order to elucidate the signal transduction pathway of vascular smooth muscle contraction induced by the activation of receptors for angiotensin II and endothelin-1, we examined whether tyrosine kinases and mitogen-activated protein (MAP) kinases are involved in the development of force of contraction in the rat aorta. Isolated aortic smooth muscles without endothelium were incubated in a modified Krebs-Henseleit solution and stimulated with angiotensin II (100 nM) or endothelin-1 (10 nM). A tyrosine kinase inhibitor genistein (10 microM) reduced the angiotensin II- and endothelin-1-induced aortic contraction, while 10 microM of daidzein (an inactive analogue of genistein) did not. The K(+) depolarization induced contraction was not attenuated by 10 microM of genistein. Selective inhibitors of MAP kinase/extracellular signal-regulated kinase (Erk) kinase (MEK) such as PD98059 [2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one] and U0126 [1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene] inhibited the angiotensin II- and endothelin-1-induced vasocontraction. The p44/42 MAP kinases were phosphorylated in cultured aortic smooth muscle cells and in physiologically contracted aortic vessels stimulated with angiotensin II and endothelin-1 for 5 min. The angiotensin II- and endothelin-1-induced phosphorylations of p44/42 MAP kinases were inhibited by PD98059 as well as U0126 in the intact aorta. These results suggest that the activation of genistein-sensitive tyrosine kinases and p44/42 MAP kinases is involved in the angiotensin II- and endothelin-1-induced rat aortic contraction. PMID- 12079692 TI - Penile erectile responses to cavernous nerve stimulation in rats are not affected by nitrate tolerance. AB - The aim of this study was to elucidate whether penile erection induced by electrical stimulation of the cavernous nerve was affected in male rats with nitrate tolerance. Nitrate tolerance to nitroglycerin was induced by oral administration of isosorbide dinitrate (ISDN) at 1000 mg/kg to rats, once or twice a day for 5 or 6 days. The rats were anesthetized with sodium pentobarbital 18-24 h after the last dosing with ISDN or its vehicle. Penile erection induced by electrical stimulation was monitored by measuring the penile diameter sonomicrometrically. After measurement of the penile erectile response, nitroglycerin (3-300 microg/kg) was intravenously (i.v.) injected into eight rats treated with the vehicle or ISDN to examine its hypotensive effect. In the vehicle-treated rats, the maximal developed penile diameter (D-max) and the duration of penile erection (T50%, period of time from the maximum erection to its 50% decline) produced by electrical stimulation were 509+/-47 microm and 14.2+/-1.7 s, respectively. On the other hand, neither D-max nor T50% in ISDN treated rats (509+/-36 microm and 13.1+/-1.3 s, respectively) was different from those in the vehicle-treated rats. However, the hypotensive effects of i.v. injected nitroglycerin were significantly attenuated in the ISDN-treated group as compared with the vehicle-treated group. It is concluded that nitrate tolerance fails to influence penile erection induced by cavernous nerve electrical stimulation in rats. PMID- 12079693 TI - Assessment of normal and ischaemic myocardium by quantitative M-mode tissue Doppler imaging. AB - This paper presents a methodology and a software package developed to quantify M mode tissue Doppler imaging (TDI), defining a number of quantitative parameters drawn from velocity and gradient curves obtained after segmenting the myocardial wall into anatomical layers. The independent clinical predictive value of these parameters to detect motion abnormalities in the presence of ischaemia was evaluated in a comparative study between a group of 17 healthy volunteers and 18 ischaemic patients. Factor analysis and stepwise logistic regression were used to assess the independent predictive value of these parameters in detecting abnormal contractility of the basal posterior segment. The statistical analysis performed has proved that any single parameter related to the gradient intensity, particularly the maximum gradient at the moment of the "e" wave, provides meaningful clinical information, achieving a rate of correct classification of 79.1% on the same data set used for the analysis. Adding additional parameters does not improve the diagnostic performance. Further testing with different settings (stress studies, other pathologies or segments) is warranted. PMID- 12079694 TI - Arterial pulse wave velocity with tissue Doppler imaging. AB - This paper describes a new noninvasive ultrasonic method for estimating pulse wave velocity (PWV), an important physical parameter for characterizing the elastic properties of the arterial walls. The method utilizes a relatively new color Doppler modality for measuring tissue motion (tissue Doppler imaging or TDI). In contrast to previously proposed methods, the TDI modality offers multiple recording sites along the artery that improve the PWV estimation considerably. The new PWV estimation method was evaluated through an in vitro setup consisting of an elastic vessel supplied with a pulsatile pump. The study concentrated on the effect of different system parameters controlling resolution, sensitivity and the amount of acquired data. It was shown that the system parameters have a significant effect on the PWV variance, whereas the PWV mean remains unchanged. It was also established that high temporal resolution is the most vital parameter for minimizing PWV variance. Finally, the new PWV estimation method was applied to a limited set of human carotid artery data sets, with good results. PMID- 12079695 TI - Functional analysis of gallbladder using three-dimensional ultrasound: preliminary results. AB - In evaluating gallbladder function, the clinical feasibility of three-dimensional (3-D) ultrasound (US) using volumetric acquisition was assessed in 30 patients: 18 patients with gallstones and 12 healthy volunteers. 3-D ultrasonography was performed in each patient before and after a fat-meal test. The ejection fraction of the gallbladder from two volumes measured by computer-aided analysis was calculated. There were significant differences in the gallbladder ejection fractions among the diseased and control groups (p < 0.0001). Compared with the results of oral cholecystograms, the ejection fraction of the gallbladder using 3 D volumetry and the grade of oral cholecystography showed good correlation in predicting the gallbladder dysfunction (gamma = 0.71, p = 0.002). Even in cases failing oral cholecystography, the gallbladder ejection fraction using 3-D US was calculated without any difficulty. Conclusively, volumetric acquisition of 3-D US enables us to evaluate gallbladder function and provides a reliable diagnostic yield superior to oral cholecystography. PMID- 12079696 TI - Ultrasonic spectral parameter characterization of apoptosis. AB - Ultrasound (US) spectral analysis methods are used to analyze the radiofrequency (RF) data collected from cell pellets exposed to chemotherapeutics that induce apoptosis and other chemicals that induce nuclear transformations. Calibrated backscatter spectra from regions-of-interest (ROI) were analyzed using linear regression techniques to calculate the spectral slope and midband fit. Two f/2 transducers, with operating frequencies of 30 and 34 MHz (relative bandwidths of 93% and 78%, respectively) were used with a custom-made imaging system that enabled the collection of the raw RF data. For apoptotic cells, the spectral slope increased from 0.37 dB/MHz before drug exposure to 0.57 dB/MHz 24 h after, corresponding to a change in effective scatterer radius from 8.7 to 3.2 microm. The midband fit increased in a time-dependent fashion, peaking at 13dB 24 h after exposure. The statistical deviation of the spectral parameters was in close agreement with theoretical predictions. The results provide a framework for using spectral parameter methods to monitor apoptosis in in vitro and in in vivo systems and are being used to guide the design of system and signal analysis parameters. PMID- 12079697 TI - Spatial variation of acoustic parameters in human skin: an in vitro study between 22 and 45 MHz. AB - This study of spatial variance of acoustic parameters was performed on eight nonfrozen samples of female abdominal skin (women 46.5 +/- 12.2 years old), obtained during plastic surgery. Intra- and interindividual variations are discussed on the basis of estimations of three acoustic parameters (slope of attenuation- beta; integrated attenuation coefficient- IAC; integrated backscattering coefficient- IBC) and one texture parameter (based on two estimators of effective density of scatterers: alpha(2) and alpha(1/2)) as a function of surface area and depth of acquisition in the frequency range 22 to 45 MHz. Values of intraindividual variations varied from 7.1% for IAC to 23.2% for IBC, and significantly decreased at a ratio between 1.2 to 2.3 when the acquisition surface area was increased from 4 mm(2) to 1 cm(2). Interindividual variations were higher than intraindividual variations, and varied from 14.2% for alpha(1/2) to 51% for IBC. The mean values (+/- SD) for all specimens combined, estimated with a large number of independent radiofrequency (RF) lines (400) and for a surface area of exploration of 4 cm(2), were 1.06 +/- 0.17 dB cm(-1) MHz( 1) for beta, 135 +/- 37 dB cm(-1) for IAC, (3.7 +/- 1.9) x 10(-2) cm(-1) sr(-1) for IBC, 1.40 +/- 0.17 scatterers/resolution cell for alpha(2) and 1.32 +/- 0.27 scatterers/resolution cell for alpha(1/2). Finally, attenuation micro(f) and backscattering coefficient sigma(b)(f) were compared to published results for the same parameters measured in human skin. PMID- 12079698 TI - Correlation of ultrasound phase with physical skull properties. AB - Noninvasive treatment of brain disorders using focused ultrasound (US) requires a reliable model for predicting the distortion of the field due to the skull using physical parameters obtained in vivo. Previous studies indicate that control of US phase alone is sufficient for producing a focus through the skull using a phased US array. The present study concentrates on identifying methods to estimate phase distortion. This will be critical for the future clinical use of noninvasive brain therapy. Ten ex vivo human calvaria were examined. Each sample was imaged in water using computerized tomography (CT). The information was used to determine the inner and outer skull surfaces, thickness as a function of position, and internal structure. Phase measurement over a series of points was obtained by placing a skull fragment between a transducer and a receiver with the skull normal to the transducer. Correlation was found between the skull thickness and the US phase shift. A linear fit of the data follows that predicted by a homogeneous skull when average speed of sound 2650 m/s was used. Large variance (SD = 60 degrees, mean = 50 degrees ) indicates the additional role of internal bone speed and density fluctuations. In an attempt to reduce the variance, the skull was first studied as a three-layer structure. Next, density-dependent bone speed fluctuation was introduced to both the single-layer and three-layer models. It was determined that adjustment of the mean propagation speeds using density improves the overall phase prediction. Results demonstrate that it is possible to use thickness and density information from CT images to predict the US phase distortion induced by the skull accurately enough for therapeutic aberration correction. In addition, the measurements provide coefficients for phase dependence on skull thickness and density that can be used in clinical treatments. PMID- 12079699 TI - On the design of a capillary flow phantom for the evaluation of ultrasound contrast agents at very low flow velocities. AB - Recently, a new imaging technology has become available that allows the evaluation of tissue perfusion using echo-contrast agents in real-time imaging: power pulse inversion imaging (PPI). Although numerous in vitro phantoms have been designed for different imaging modalities in ultrasound (US), there is a need for a phantom that mimics microcirculation and allows, in particular, the assessment of contrast replenishment kinetics following US-induced destruction of microbubbles using the new method. We, therefore, designed a new capillary flow phantom that takes the requirements of the new US imaging techniques and the physical properties of microbubbles into account and serves flow velocities in the range of microcirculation (1 to 10 mm/s). PPI studies were performed in the newly designed phantom. The contrast agent used was AF0150. We studied homogeneity of contrast distribution within the capillary phantom, constancy of contrast infusion, the dose-effect relationship and, finally, the feasibility of flow assessment using the method of contrast replenishment following US-induced microbubble destruction in a flow velocity range of 2.1 to 9.45 mm/s. Analysis of the replenishment kinetics was performed using the mathematical model f(t) = A(1 e(-beta t)), with A representing the blood volume and beta the microbubble velocity. The new capillary phantom allowed homogeneous contrast opacification within the perfused capillaries independently of the flow. Constancy of signal intensity was achieved over a time period of almost 2 h, indicating constant contrast delivery. A strong linear correlation between the PPI signal and the contrast dose was found (r = 0.998). Analysis of the replenishment parameters revealed a strong linear relationship between parameter beta and flow (r = 0.994) as well as A * beta and flow (r = 0.984) in the observed flow range. The newly designed perfusion phantom for the evaluation of echo-contrast replenishment kinetics fulfills, at very low flow velocities, important prerequisites such as constancy of contrast delivery, homogeneity of contrast signals, linear dose effect relation and minimal attenuation. Thus, the new phantom allows standardized analysis of contrast replenishment kinetics using real-time perfusion imaging techniques at flow velocities comparable to those of the microcirculation. PMID- 12079701 TI - Reduction of coherent scattering noise with multiple receiver Doppler. AB - Doppler ultrasound (US) velocity estimates are inherently subject to error as a result of both Doppler ambiguity and coherent scattering. The coherent scattering error is a result of changes in the phase of the returned echo as particles enter and leave the sample volume. This phase depends on the distance from the transmitter to the scatterer and then to the receiver. This distance, in turn, depends on the angle of the receiver. A numerical simulation has been used to determine whether velocity estimates obtained from receiver probes at different angles are independent of one another. If so, then it is possible to obtain an improved velocity estimate from the combination of several receivers at different angles. The simulation results show that the cross-correlation between velocity estimates is reduced to 0.3 when receiver probes are oriented 5 degrees apart. These results suggest a new Doppler method that can significantly reduce velocity estimation error. PMID- 12079700 TI - Experimental characterization of fundamental and second harmonic beams for a high frequency ultrasound transducer. AB - In the diagnostic frequency range, nonlinear imaging has been shown to improve image contrast and decrease artefacts. The extension of these techniques to high frequency imaging (>15 MHz) was investigated. The second harmonic beam at 40 MHz of a high-frequency focused transducer (aperture 6 mm, focal distance 10 mm, f number 1.67) was measured experimentally in water, in transmission and pulse echo, and compared with the fundamental beams at 20 MHz and 40 MHz. Measurements were performed at peak negative pressures of 0.8 to 4.7 MPa. Transmission measurements were performed with a custom hydrophone with a 25microm spot size to limit beam averaging. Over the range of peak negative pressures, the transmitted harmonic (40 MHz) beam had an average lateral beam width (-3 dB) of 77 microm and an average depth-of-field of 0.93 mm, whereas the fundamental beam had a corresponding beam width of 137 microm and a depth-of-field of 1.59 mm. The harmonic beam showed a 3-dB decrease in side lobe levels. Preliminary second harmonic images of mouse tissue in vitro are presented and compared to fundamental imaging at 20 and 40 MHz. PMID- 12079702 TI - High-frequency sound field and bubble formation in a rat decompression model. AB - High-frequency sound might cause bubble enlargement by rectified diffusion. The purpose of the present study was to investigate gas bubble formation in the immersed diving animal during exposure to high-frequency sound. Anaesthetised rats were subjected to a simulated diving profile while immersed inside a hyperbaric chamber. An acoustic beacon (pinger) was placed ventral to the animal's abdomen, transmitting at an intensity of 208.9 dB re 1 micro Pa and a frequency of 37 kHz. Six groups of eight animals were included in the study as in Table 1, breathing air (n = 4) or Nitrox 72/28 (n = 2), at a depth of 0 m, 30 m or 40 m. Immediately after decompression, the intestinal mesenterium was imaged, and frames were acquired digitally. The number of bubbles and their radii were analysed and compared among the groups. The mean bubble density for group 1 was 1.35 +/- 0.18 bubbles/mm(2), significantly higher when compared with the other groups (p < 0.0001). The average bubble radius for groups 1 and 2 was similar (12.57 +/- 4.1 and 10.63 +/- 1.8 microm, respectively), but significantly larger than in the other groups (p < 0.0002). The percentage of bubbles with a radius greater than 50 microm was significantly higher in group 1 (p < 0.0001). The results suggest that commercially available underwater pingers might enhance bubble growth during deep air diving. PMID- 12079703 TI - The role of stress waves and cavitation in stone comminution in shock wave lithotripsy. AB - Using an experimental system that mimics stone fragmentation in the renal pelvis, we have investigated the role of stress waves and cavitation in stone comminution in shock-wave lithotripsy (SWL). Spherical plaster-of-Paris stone phantoms (D = 10 mm) were exposed to 25, 50, 100, 200, 300 and 500 shocks at the beam focus of a Dornier HM-3 lithotripter operated at 20 kV and a pulse repetition rate of 1 Hz. The stone phantoms were immersed either in degassed water or in castor oil to delineate the contribution of stress waves and cavitation to stone comminution. It was found that, while in degassed water there is a progressive disintegration of the stone phantoms into small pieces, the fragments produced in castor oil are fairly sizable. From 25 to 500 shocks, clinically passable fragments (< 2 mm) produced in degassed water increases from 3% to 66%, whereas, in castor oil, the corresponding values are from 2% to 11%. Similar observations were confirmed using kidney stones with a primary composition of calcium oxalate monohydrate. After 200 shocks, 89% of the fragments of the kidney stones treated in degassed water became passable, but only 22% of the fragments of the kidney stones treated in castor oil were less than 2 mm in size. This apparent size limitation of the stone fragments produced primarily by stress waves (in castor oil) is likely caused by the destructive superposition of the stress waves reverberating inside the fragments, when their sizes are less than half of the compressive wavelength in the stone material. On the other hand, if a stone is only exposed to cavitation bubbles induced in SWL, the resultant fragmentation is much less effective than that produced by the combination of stress waves and cavitation. It is concluded that, although stress wave-induced fracture is important for the initial disintegration of kidney stones, cavitation is necessary to produce fine passable fragments, which are most critical for the success of clinical SWL. Stress waves and cavitation work synergistically, rather than independently, to produce effective and successful disintegration of renal calculi in SWL PMID- 12079705 TI - Optimum intensities of ultrasound for PGE(2) secretion and growth of osteoblasts. AB - This study compared the effects of different intensity ultrasound (US) on osteoblasts in the far-field model with effects of the near-field model from the literature, to understand the relations between prostaglandin E(2) (PGE(2)) and osteoblast growth. We used an in vitro model to investigate the effects of 1-MHz, pulsed 1:4, and five different spatial-average temporal-peak intensity (150, 300, 600, 1200 and 2400 mW/cm(2)) US stimulations in far-field exposure (240 mm) on osteoblasts for 15 min. Optimum intensity in this study was 600 mW/cm(2), and cell density and PGE(2) secretion could be significantly stimulated at this intensity. This research may indicate that the growth of osteoblasts by US stimulation was, at least partly, due to increases in the synthesis and secretion of PGE(2). This well-controlled model can lead to further research on the biologic mechanisms for US. PMID- 12079704 TI - Effects of dissolved gases and an echo contrast agent on apoptosis induced by ultrasound and its mechanism via the mitochondria-caspase pathway. AB - Human histiocytic lymphoma U937 cells were exposed to continuous 1-MHz ultrasound (US) for therapeutic use, (0 approximately 6.5 W/cm(2) (I(SPTA)). Apoptosis and its related end points were examined by flow cytometry. Fraction of cells with low mitochondria membrane potential were observed after sonication and significant superoxide and peroxide formation, increased activity of caspase-3, and DNA fragmentation revealed biochemically, were also found. The fraction of early apoptosis and secondary necrosis increased with the incubation time after sonication. Early apoptosis observed at 6 h after sonication reached its maximum at 2 min of sonication and gradually decreased. On the other hand, secondary necrosis increased with the duration of sonication. When the effects of dissolved gases, Ar, N(2), O(2), air, N(2)O and CO(2), on free radical formation due to inertial cavitation were investigated by electron spin resonance (ESR) spin trapping, formation of hydroxyl radicals and hydrogen atoms was found in solutions saturated with Ar, N(2), O(2) and air, but not with N(2)O and CO(2). Apoptosis induced by US was also dependent on the dissolved gases in the order Ar = N(2) = O(2) = air >> N(2)O = CO(2) approximately 0. These results suggest that US-induced apoptosis, which is mitochondria-caspase dependent, was linked to inertial cavitation. However, quantities of free radicals did not influence the fraction of early apoptosis and secondary necrosis. When the cells were sonicated in the presence of an echo contrast agent, Levovist; synergistic enhancement of secondary necrosis induced by US was observed at concentrations of more than 20 mg/mL. In contrast, an additive increase of early apoptosis was observed in the combined treatments. These results suggest that Levovist; acting as cavitation nuclei, enhances secondary necrosis induced by US due to an increase in the membrane damage. PMID- 12079706 TI - Potential of gold-bound microtubules as a new ultrasound contrast agent. AB - Contrast agents based on gas-filled microspheres share the problem of time limited opacification due to low stability of microbubbles. The aim of this study was to test if gold-bound microtubules provide backscattering that allows microtubules to be potentially useful as an ultrasound (US) contrast agent. Gold colloids were immobilized on protein microtubule walls. Latex balloons were filled with gold-bound microtubules or conventional left heart contrast agent and were ultrasonographically imaged in fundamental and harmonic modes. Feasibility of anti-beta-tubulin antibody conjugation to gold-bound microtubules was confirmed using immune fluorescence analysis. Gold particles were successfully bound to microtubules. Contrast intensities in latex balloons filled with gold bound microtubules (141 +/- 35) were comparable to those with Levovist (180 +/- 35) and did not decrease significantly during continuous US imaging for 20 min (135 +/- 34 vs. Levovist 5.0 +/- 2.0). Anti-beta-tubulin antibodies were successfully conjugated to gold-bound microtubules. Gold-bound microtubules provide a persistent contrast effect, suggesting their use as an ultrasonic contrast agent with the feasibility of antibody conjugation. PMID- 12079708 TI - Sports ability in young men and the incidence of cardiovascular disease. AB - PURPOSE: For physical activity to reduce the risk of cardiovascular disease, it must be sustained throughout life. We sought to determine the relation between the ability of young adults in different sports and their continued physical activity in midlife, and subsequent cardiovascular disease. SUBJECTS AND METHODS: Baseline self-reported ability in tennis, golf, football, baseball, and basketball was assessed in a cohort study involving 1019 male medical students (median age, 22 years). Physical activity and sports participation were assessed 22 years later. The incidence of cardiovascular disease was assessed during a median follow-up of 40 years, using annual questionnaires, medical records, and death certificates. RESULTS: Subjects with high ability in tennis as young adults had the highest participation in the sport in midlife (median age, 48 years). In the midlife questionnaire administered in 1978, 33% reported playing tennis within the past week and 51% within the past year. Tennis was the only sport in which a higher ability during medical school was associated with a lower risk of cardiovascular disease. After adjustment for father's occupation, parental incidence of cardiovascular disease, serum cholesterol level, cigarette smoking, body mass index, and hypertension during follow-up, the relative hazard of developing cardiovascular disease was 0.56 (95% confidence interval [CI]: 0.35 to 0.89) in the high-ability group and 0.67 (95% CI: 0.47 to 0.96) in the low ability group, compared with the no-ability group. CONCLUSION: Our results support the association between sustained activity in aerobic sports and a lower risk of cardiovascular disease. Sustainability of activity should be considered when developing physical education programs for young adults. PMID- 12079709 TI - The effects of comorbid conditions on the outcomes of patients undergoing peritoneal dialysis. AB - PURPOSE: Dialysis patients frequently have comorbid conditions. We examined the effects of age and comorbid conditions on technique failure (i.e., transfer to hemodialysis), death, hospital costs, and kidney transplantation in patients treated with peritoneal dialysis. METHODS: We studied 97 patients who began peritoneal dialysis from January 1, 1993, to December 31, 1998, at the University of Pittsburgh outpatient dialysis unit. Demographic characteristics, comorbid conditions, and outcomes were determined by reviewing the Medical Archival Retrieval System database and outpatient records. Because the comorbidity (Charlson) score was colinear with age, we used a modified version of the score without an age component. Low, moderate, and high comorbidity groups were defined based on the 33rd and 66th percentiles of the comorbidity score. RESULTS: In multivariate-adjusted models, each decade increase in age was associated with an increased risk of death (hazard ratio [HR] = 1.7; 95% confidence interval [CI]: 1.1 to 2.5) and technique failure (HR = 1.5; 95% CI: 1.0 to 2.3). High (versus low) comorbidity was associated with an increased risk of death or technique failure (HR = 3.5; 95% CI: 1.0 to 12) and significantly higher average inpatient costs. There were no differences in age or comorbidity score between patients who transferred to hemodialysis and those who died. CONCLUSION: Patients who are older and more ill have a greater risk of death and of transfer to hemodialysis from peritoneal dialysis. PMID- 12079710 TI - The effects of time-to-surgery on mortality and morbidity in patients following hip fracture. AB - PURPOSE: There is a perception that the standard of care is to repair hip fractures surgically within 24 hours of hospitalization. However, it is unclear whether this reduces mortality or morbidity. SUBJECTS AND METHODS: We performed a retrospective study in consecutive hip fracture patients, aged 60 years or older, who underwent surgical repair. Patients with metastatic cancer, trauma, or a fracture occurring >48 hours before admission were excluded. The primary outcome was long-term (up to 18 years) mortality. Secondary outcomes included 30-day mortality and decubitus ulcers, serious bacterial infections, myocardial infarction, and thromboembolism. Analyses were adjusted for medical conditions; the comparison group comprised patients who underwent surgery for hip fracture repair within 24 to 48 hours because there were no patients with active medical problems who underwent surgery within 24 hours. RESULTS: Of the 8383 patients, surgery was delayed for more than 24 hours in 2464 patients (29%) for medical reasons and in 1341 patients (16%) without active medical problems. Compared with those who underwent surgery 24 to 48 hours after admission to the hospital, patients who underwent surgery more than 96 hours after admission did not have increased long-term mortality (hazard ratio = 1.07; 95% confidence interval [CI]: 0.95 to 1.21), although the risk of decubitus ulcer was increased (odds ratio = 2.2; 95% CI: 1.6 to 3.1). There were no associations between time-to-surgery and the other secondary outcomes. CONCLUSION: Time-to-surgery in hip fracture patients was not associated with short- or long-term mortality after adjusting for active medical problems. Other than increasing the risk of decubitus ulcer formation, waiting did not appear to affect patients' outcomes adversely. PMID- 12079711 TI - Appetite suppressants and valvular heart disease in a population-based sample: the HyperGEN study. AB - PURPOSE: Previous studies of the association between the use of appetite suppressants and valvular heart disease have not accounted for the effects of valvular structure and aortic root diameter, which are associated with obesity. We assessed whether the use of the appetite suppressants fenfluramine/dexfenfluramine, either alone or with phentermine, was associated with aortic regurgitation while adjusting for these variables. SUBJECTS AND METHODS: The sample included 2524 adult participants in the population-based Hypertension Genetic Epidemiology Network study. Information regarding current drug use was assessed during a clinical examination. Medication use was continued at the time of echocardiographic study. Expert readers blinded to current therapy read echocardiograms centrally at Cornell Medical Center. Analyses of the associations between use of fenfluramine/dexfenfluramine (alone or with phentermine) and aortic regurgitation adjusted for potential confounders, including aortic root dilatation and valve fibrocalcification. RESULTS: Nineteen participants, all of whom had hypertension, were being treated with fenfluramine or dexfenfluramine (5 on these agents alone, 14 also with phentermine). Aortic regurgitation was present in 32% (n = 6) of those taking fenfluramine or dexfenfluramine versus 6% (162/2505) of remaining subjects (P = 0.001). In multivariate-adjusted analyses, treatment with fenfluramine or dexfenfluramine was associated with aortic regurgitation (odds ratio [OR] = 4.9; 95% confidence interval [CI]: 1.7 to 14) and aortic fibrocalcification (OR = 5.2; 95% CI: 1.9 to 15). CONCLUSION: In a population-based sample, use of fenfluramine or dexfenfluramine, alone or in combination with phentermine, was associated with aortic regurgitation independent of aortic dilatation or fibrocalcification. PMID- 12079712 TI - B-type natriuretic peptide as a marker of the effects of enalapril in patients with heart failure. AB - BACKGROUND: A-Type and B-type natriuretic peptides are cardiac hormones whose circulating levels reflect the severity of heart failure. It is not known how plasma levels of these hormones respond to changes in cardiac function that occur as a result of treatment with angiotensin-converting enzyme (ACE) inhibitors. METHODS: Enalapril was administered at 5 mg/d for 3 months in 24 patients with chronic heart failure, and for the next 3 months at 15 mg/d in the high-dose group (n = 12) and 5 mg/d in the low-dose group (n = 12). We measured plasma levels of A-type or B-type natriuretic peptides, as well as conventional measures of cardiac function, such as the cardiothoracic ratio, left ventricular end diastolic volume, and percent fractional shortening. RESULTS: Mean (+/- SD) plasma levels of both hormones decreased promptly after 2 weeks of therapy (A type natriuretic peptide: 140 +/- 107 pg/mL to 81 +/- 68 pg/mL, P = 0.01; B-type natriuretic peptide: 305 +/- 278 pg/mL to 190 +/- 178 pg/mL, P = 0.01). These reductions were sustained throughout therapy. In contrast, the cardiothoracic ratio, left ventricular end-diastolic dimension, and percent fractional shortening had not changed significantly after 3 months of treatment, although improvements were seen after 6 months of treatment. After 6 months, plasma levels of both A-type and B-type natriuretic peptides were significantly lower in the high-dose group than in the low-dose group (A-type natriuretic peptide: 48 +/- 25 pg/mL vs. 57 +/- 27 pg/mL, P = 0.01; B-type natriuretic peptide: 78 +/- 58 pg/mL vs. 139 +/- 61 pg/mL, P = 0.005), whereas no significant differences were observed in the other measures of cardiac function. CONCLUSION: Plasma levels of A-type and B-type natriuretic peptides appear to be more sensitive markers of heart failure than conventional echocardiographic parameters and cardiothoracic ratios. Measurement of these hormones might be useful for monitoring the effects of treatment with ACE inhibitors. PMID- 12079713 TI - The use of recombinant thyrotropin in the follow-up of patients with differentiated thyroid cancer. AB - Many clinicians care for patients who have been treated for differentiated thyroid cancer. Recombinant thyrotropin, which stimulates iodine uptake in thyroid tissue, is a safe and effective diagnostic agent for those patients who require radioiodine scanning for routine follow-up. The combination of a whole body radioiodine scan and a serum thyroglobulin measurement can identify virtually all patients with distant metastatic disease. A serum thyroglobulin >2 ng/mL and/or a positive whole body scan after recombinant thyrotropin stimulation suggest residual thyroid tissue or neoplastic disease. The use of recombinant thyrotropin has fewer adverse effects than does the alternative, which is withdrawal of thyroid hormone replacement, although nausea and headache have been reported. However, recombinant thyrotropin is expensive. PMID- 12079714 TI - Demographic differences in the development of lupus nephritis: a retrospective analysis. PMID- 12079715 TI - Cases from the medical grand rounds of the Osler Medical Service at Johns Hopkins University. AB - A 77-year-old man was in good health until he complained of fatigue 3 weeks before presentation. Two weeks before admission, he developed gradually worsening shortness of breath. One week before admission, he developed a cough that initially was nonproductive but later was associated with hemoptysis.His past medical history was remarkable for a history of colon cancer (Dukes' stage III), for which he underwent a hemicolectomy and treatment with adjuvant chemotherapy in 1993. He had a myocardial infarction in 1986 and underwent coronary artery bypass surgery in 1999. He also had a history of hypertension, type 2 diabetes, and gout. He smoked in the past but had stopped more than 30 years ago.He was initially evaluated by his primary care physician, who noted that he complained of diaphoresis but denied fevers, chills, or contact with others who were ill. His physical examination was remarkable for bilateral crackles that were more pronounced on the right. A chest radiograph demonstrated bilateral pulmonary infiltrates (Figure 1). He was treated with amoxicillin. The next day, however, his physician noted that his dyspnea had worsened and that his oxygen saturation on room air was poor. He was therefore admitted for further evaluation. The amoxicillin was discontinued, and he was treated with levofloxacin, followed by ceftriaxone and azithromycin as his pulmonary status continued to deteriorate. He received intravenous diuretic agents, which failed to improve his respiratory status. During the initial phase of hospitalization, he was anemic, with a hematocrit of 21.3%. His serum creatinine level, which had been 1.0 mg/dL in 1999, was now 2.5 mg/dL. Urinalysis was remarkable for the presence of numerous red blood cells. His oxygen requirement increased, and he eventually required a 100% nonrebreather mask. A computed tomographic scan of the chest demonstrated prominent alveolar opacities throughout the right upper, middle, and lower lobes, with similar opacities in the left upper and left lower lobes (Figure 2). An echocardiogram showed an ejection fraction of 50%, as well as mild mitral regurgitation. Serologies were remarkable for an antinuclear antibody titer of 1:320 and a P-antineutrophil cytoplasmic antibody (P-ANCA) titer of greater than 1:320. C-ANCA was negative. Anti-glomerular basement membrane and anti-human immunodeficiency virus antibodies were undetectable. PMID- 12079716 TI - Tennis, anyone? On the value of sustainable, vigorous physical activity and long term studies. PMID- 12079717 TI - What is the best treatment for end-stage renal disease? PMID- 12079718 TI - False-positive HIV-1 ELISA in patients with hepatitis B. PMID- 12079719 TI - Tuberculous esophageal ulcer as the mode of presentation of Pott's disease (tuberculous spondylitis). PMID- 12079720 TI - Intrathoracic extramedullary hematopoiesis resembling posterior mediastinal tumor. PMID- 12079721 TI - Terbinafine hepatotoxicity resulting in chronic biliary ductopenia and portal fibrosis. PMID- 12079722 TI - Asymptomatic constitutional macrothrombocytopenia among West Bengal blood donors. PMID- 12079723 TI - Professing and living the oath: teaching medicine as a profession. PMID- 12079724 TI - Morphology of the ventral frontal cortex in schizophrenia: relationship with social dysfunction. AB - BACKGROUND: Studies have reported premorbid as well as postonset social dysfunction in schizophrenia. This impairment has also been observed to emerge after lesions in the ventral aspect of the frontal cortex (i.e., straight gyrus and orbitofrontal cortex). METHODS: Magnetic resonance imaging scans were obtained from 45 male patients with schizophrenia and 45 matched control subjects. Cortical gray matter volume and surface area were determined for the ventral frontal cortex (VFC), subdivided into the orbitofrontal cortex (OFC) and the straight gyrus (SG). RESULTS: The global measures of gray matter volume and surface area in the VFC was not significantly different between patients and control subjects; however, there was a regional difference, with the right SG volume and surface area being smaller in patients compared with control subjects. Volume of the VFC had an inverse correlation with measurements of both premorbid and postdiagnosis social function. The smaller the gray matter in these regions, the greater the social dysfunction. There was no relationship between morphology of this brain region and any other clinical variable. CONCLUSIONS: Morphology of the VFC is directly related to abnormal social function in schizophrenia, including measures of social dysfunction that predate the onset of illness. PMID- 12079726 TI - Paracingulate morphologic differences in males with established schizophrenia: a magnetic resonance imaging morphometric study. AB - BACKGROUND: Our previous work on sulcal-gyral brain morphology in healthy volunteers revealed that males were characterized by greater cortical folding in the left versus right anterior cingulate cortex. Given the evidence showing an absence or reversal of normal anatomical asymmetries in patients with schizophrenia, the current study examined the anterior cingulate cortex sulcal gyral patterns in patients with schizophrenia. METHODS: Using high-resolution magnetic resonance imaging, we examined anterior cingulate cortex surface morphology in a group of 55 patients with established schizophrenia and 75 healthy controls. All subjects were male and right-handed. Depending on the presence of a paracingulate sulcus and its antero-posterior extent, three types of anterior cingulate cortex sulcal patterns were identified: "prominent," "present," and "absent." Measures of overall cerebral hemispheric folding were used as independent variables and as covariates to ascertain the specificity of the findings to the anterior cingulate cortex. RESULTS: Examination of anterior cingulate cortex morphology showed that, compared with controls, patients with schizophrenia lacked the leftward anterior cingulate cortex sulcal asymmetry, which was explained by reduced folding in the left anterior cingulate cortex. These differences were over and above differences in cortical folding across the entire left hemisphere. CONCLUSIONS: These findings suggest that, in male patients with schizophrenia, there is a disturbance in the neurodevelopment of the left anterior cingulate cortex, as well as a more general aberration of left hemisphere development. PMID- 12079725 TI - Frontal white matter microstructure, aggression, and impulsivity in men with schizophrenia: a preliminary study. AB - BACKGROUND: Aggression and impulsivity may involve altered frontal white matter. METHODS: Axial diffusion tensor images were acquired in 14 men with schizophrenia using a pulsed gradient, double spin echo, echo planar imaging method. White matter microstructural measures (fractional anisotropy and trace) were calculated from these data. Regions of interest were placed in frontal white matter on four slices. Impulsivity was measured using the Motor Impulsiveness factor of the Barratt Impulsiveness Scale. Aggressiveness was measured using the Assaultiveness scale of the Buss Durkee Hostility Inventory and the Aggression scale of the Life History of Aggression. RESULTS: Lower fractional anisotropy in right inferior frontal white matter was associated with higher motor impulsiveness. Higher trace in these regions was associated with aggressiveness. CONCLUSIONS: Inferior frontal white matter microstructure was associated with impulsivity and aggression in men with schizophrenia. These results implicate frontal lobe dysfunction in aggression and certain aspects of impulsivity. PMID- 12079727 TI - Motor cortical excitability in schizophrenia. AB - BACKGROUND: Transcranial magnetic stimulation (TMS) provides a method to examine cortico-cortical motor excitability and hemispheric asymmetry in unmedicated and medicated schizophrenia patients. METHODS: Fourteen right-handed schizophrenia patients (seven on conventional neuroleptics and seven medication-free) were compared with seven right-handed, age- and gender-matched normal control subjects. Motor threshold for induction of motor-evoked potentials (MEPs) and bihemispheric intracortical inhibition and facilitation were measured with single pulse and paired-pulse TMS. RESULTS: Medicated patients showed an approximately 5% higher motor thresholds in both hemispheres than unmedicated patients and control subjects. Normal control subjects had a nearly 10% higher threshold for the left than the right hemisphere, whereas the opposite was true for the patient groups (5-10% higher threshold on the right than the left). Medicated patients showed significantly decreased intracortical inhibition relative to unmedicated patients and control subjects. This difference was more pronounced for the right than for the left hemisphere. CONCLUSIONS: Treatment with conventional neuroleptics is associated with increased motor threshold and decreased intracortical inhibition, whereas unmedicated patients did not differ from normal control subjects on these measures; however, schizophrenia may be characterized by a reversed pattern of interhemispheric corticospinal excitability. PMID- 12079728 TI - Alterations in tonotopy and auditory cerebral asymmetry in schizophrenia. AB - BACKGROUND: Deficits in basic auditory perception have been described in schizophrenia. Previous electrophysiologic imaging research has documented a structure-function disassociation in the auditory system in schizophrenia. This study examines whether the most fundamental level of auditory cortical organization, tonotopy, is altered in schizophrenia. METHODS: The tonotopic organization for five tone frequencies in 19 patients with schizophrenia and 22 comparison subjects was evaluated using magnetoencephalography. Auditory evoked magnetic field dipole locations were examined for the N100m component for each frequency. RESULTS: The expected linear relationship between depth and frequency was found in the comparison subjects but not in the schizophrenia group (p <.004). In addition, normal anterior-posterior asymmetry of the N100m was found to be reduced at all five stimulation frequencies employed in the study (p <.04). No relationships between clinical symptom ratings and either tonotopy or asymmetry were observed. CONCLUSIONS: This finding suggests that the tonotopic organization of the auditory cortex in schizophrenia is disturbed and may help explain the relatively poor behavioral performance of schizophrenia patients on simple frequency discrimination tasks. Alterations in fundamental sensory organization may underlie or interact with higher order cognitive mechanisms to produce changes in cognitive task performance. PMID- 12079729 TI - Genome scan for susceptibility loci for schizophrenia and bipolar disorder. AB - BACKGROUND: Despite the widely accepted view that schizophrenia and bipolar disorder represent independent illnesses and modes of inheritance, some data in the literature suggest that the diseases may share some genetic susceptibility. The objective of our analyses was to search for vulnerability loci for the two disorders. METHODS: A genomewide map of 388 microsatellite DNA markers was genotyped in five schizophrenia and three bipolar disorder Austrian families. Linkage analyses was used to compute the usual parametric logarithm of the likelihood of linkage (LOD) scores and nonparametric linkage analysis (NPL scores Z(all)) was used to assess the pattern of allele sharing at each marker locus relative to the presence of the disease (GENEHUNTER). Affected status was defined as severe affective disorder or schizophrenia. RESULTS: Across the genome, p values associated with NPL scores resulted in evidence (i.e., p <.0007) for linkage at marker D3S1265 on chromosome 3q (NPL score Z (all) = 3.74, p =.0003). Two other markers (on 3q and 6q) showed p values of <.01. CONCLUSIONS: We detected a potential susceptibility locus for bipolar disorder and schizophrenia on chromosome 3q, which has not been reported previously. The possibility of a false positive result has to be taken into account. Our data suggest shared loci for schizophrenia and bipolar affective disorders and are consistent with the continuum model of psychosis. PMID- 12079730 TI - A placebo controlled trial of bupropion for smoking cessation in schizophrenia. AB - BACKGROUND: Schizophrenic patients have high rates of cigarette smoking compared with the general population. We compared sustained-release (SR) bupropion with placebo for smoking cessation in patients with schizophrenic disorders. We also examined how antipsychotic class predicts smoking cessation outcomes with bupropion. METHODS: Thirty-two subjects meeting DSM-IV criteria for schizophrenia or schizoaffective disorder and nicotine dependence were randomized to bupropion SR (BUP, 300 mg/day) or placebo (PLA). Outcomes included treatment retention, smoking abstinence rates, expired breath carbon monoxide (CO) levels, psychotic symptoms, and medication side effects. RESULTS: Bupropion significantly increased trial endpoint 7-day point prevalence smoking abstinence rates compared with placebo [BUP, 8/16 (50.0%), PLA, 2/16 (12.5%); chi(2) = 5.24, df = 1, p <.05], and reduced CO levels during the trial [Medication x Time interaction; Z = 3.09, p <.01]. Positive schizophrenia symptoms were not altered by BUP, but negative symptoms were significantly reduced. Atypical antipsychotic drug treatment enhanced smoking cessation responses to BUP. Major side effects were dry mouth, gastrointestinal symptoms, headache, and insomnia. CONCLUSIONS: Our results suggest that 1) BUP enhances smoking abstinence rates compared with PLA in nicotine-dependent schizophrenic smokers; 2) BUP is well-tolerated and safe for use in these patients; and 3) atypical antipsychotics may enhance smoking cessation outcomes with BUP. PMID- 12079731 TI - An application of the revised CONSORT standards to FDA summary reports of recently approved antidepressants and antipsychotics. AB - BACKGROUND: Recent years have seen the introduction of many antidepressants and antipsychotics. Typically, efficacy of these agents is based on published positive clinical trials; however, the Food and Drug Administration (FDA) compiles summary reports of all pivotal randomized clinical trials, which are available for public review under the Freedom of Information Act. They are rarely accessed but may have public health utility. Thus, we assessed these reports to evaluate if the clinical data can be used for comparing psychotropics. METHODS: We examined FDA summary reports of 58 pivotal trials from 10 antidepressants and 7 pivotal trials from three antipsychotics approved in the United States between 1985 and 1998. We evaluated each FDA report based on criteria derived from the recently revised Consolidated Standards of Reporting Trials (CONSORT standards) proposed by the Journal of the American Medical Association to improve randomized clinical trial reports. RESULTS: Overall, the FDA reports of antidepressant and antipsychotic trials adequately met 38.1% of the CONSORT criteria. Among the antidepressant reports, 36.7% adequately met CONSORT criteria compared with 49.0% among the antipsychotic reports. The proportions meeting CONSORT criteria have increased over the past 15 years; 21.0% for those approved between 1985 and 1989, 37.8% for those between 1990 and 1994, and 40.5% for those between 1995 and 1998. CONCLUSIONS: The large and inclusive public domain FDA summary reports may provide balanced efficacy clinical trial data. With minor changes in the preparation of reporting by using CONSORT or similar guidelines, the data would be even more valuable. PMID- 12079732 TI - Increased basal ganglia volumes in velo-cardio-facial syndrome (deletion 22q11.2). AB - BACKGROUND: This study evaluated differences in caudate volumes in subjects with velo-cardio-facial syndrome due to a 22q11.2 (22qDS) deletion. Because psychosis is observed in 30% of adult subjects with 22qDS, this neurogenetic disorder could represent a putative model for a genetically mediated subtype of schizophrenia. METHODS: Caudate volumes were measured on high-resolution magnetic resonance images in 30 children and adolescents with 22qDS and 30 gender- and age-matched normal comparison subjects. RESULTS: Caudate head volumes were increased in the 22qDS group independent of neuroleptic medications. Subjects with 22qDS also displayed an abnormal pattern of asymmetry in the anterior caudate, with left side greater than right. CONCLUSIONS: Alterations in the basal ganglia circuitry have been implicated in learning, cognitive, and behavioral problems in children and therefore could be involved in the expression of the neurobehavioral phenotype expressed by subjects with 22qDS. Abnormal caudate volume is a neurodevelopmental feature shared with schizophrenia, further establishing 22qDS as a potential neurodevelopmental model for this disorder. PMID- 12079733 TI - Trichomonas gallinae: a possible contact-dependent mechanism in the hemolytic activity. AB - The in vitro hemolytic activity of Trichomonas gallinae was investigated. The parasite was tested against human erythrocytes of groups A, B, AB, and O, and against erythrocytes of six adult animals of different species (rabbit, rat, chicken, horse, bovine, and sheep). Results showed that T. gallinae lysed all human erythrocytes groups, as well as rabbit, rat, chicken, horse, bovine and sheep erythrocytes. No hemolysin released by the parasites could be identified. Hemolysis did not occur with trichomonad culture supernatants, with sonicated extracts of T. gallinae, or with killed organisms. The scanning electron microscopy (SEM) showed that the erythrocytes adhered to the parasite surface and were phagocytosed. These observations suggest that the contact between T. gallinae and erythrocytes may be an important mechanism in the injury caused to the erythrocytes. The hemolytic activity of T. gallinae may be an efficient means of obtaining nutrients for the parasite and allow the investigation of the mechanism used by T. gallinae to damage cellular membranes. PMID- 12079734 TI - Molecular characterisation of Babesia canis canis and Babesia canis vogeli from naturally infected European dogs. AB - The morphologically small Babesia species isolated from naturally infected dogs in Europe, Japan, and US are described as Babesia gibsoni despite the fact that molecular techniques show that they should be assigned to two or three separate taxons. The morphologically large Babesia isolated from dogs in Europe, Africa, and US were generally classified as B. canis until it was proposed to distinguish three related, albeit genetically distinct subspecies of this genus, namely B. canis canis, B. canis rossi, and B. canis vogeli. The insight into the molecular taxonomy of canine piroplasms is, however, limited because only partial small subunit ribosomal RNA (ssrRNA) sequence data exist for two species from the B. canis group. In this work, we molecularly characterised natural Babesia infections in 11 dogs from Croatia, France, Italy, and Poland. These infections were diagnosed as caused by B. canis canis and B. canis vogeli based on the analysis of the complete sequence of the ssrRNA genes. Phylogenetic analysis confirmed that the large Babesia species of dogs belong the to the Babesia sensu stricto clade, which includes species characterised by transovarial transmission in the tick vectors and by exclusive development inside the mammalian host erythrocytes. The new data facilitate the reliable molecular diagnosis of the subspecies of B. canis. PMID- 12079735 TI - p38-avidity-ELISA: examination of herds experiencing epidemic or endemic Neospora caninum-associated bovine abortion. AB - An enzyme-linked immunosorbent assay (ELISA) was established to measure the avidity of bovine IgG directed against p38, a surface antigen (NCSRS2) of Neospora caninum tachyzoites. In the sera of dams intravenously infected with N. caninum NC-1, the p38-specific avidity increased from initially below 40% up to avidity indices between 50 and 80% after days 23 and 91 p.i. The p38-avidity ELISA was used to examine various herds that had experienced endemic and epidemic N. caninum-associated bovine abortions. In herds with endemic abortion, generally high avidity indices of N. caninum-specific IgG were detected. This finding and the observation of an association between the seropositivity of dams and that of their offspring suggested a predominantly vertical transmission of the parasite among the animals of these herds, thus indicating chronic infection of these cattle. In contrast, for herds experiencing epidemic abortion: (i) an association regarding seropositivity of dams and their daughters could not be shown and (ii) a generally low avidity of p38-specific IgG in the sera from aborting dams was determined. This indicates recent postnatal transmission of N. caninum in these herds. A linear regression model explaining the avidity of p38-specific IgG was significantly influenced by the time span between sampling and the occurrence of the first abortion of the epidemic in herds with an N. caninum-associated abortion storm. Another factor significantly contributing to the model was the proportion of dams at risk that had aborted in the herds (i.e. the severity of the abortion epidemic). A possible explanation for this observation is that herds experiencing heavy abortion are sampled earlier after the onset of the epidemic than others that have a less severe abortion storm. PMID- 12079736 TI - Serological survey of Neospora caninum and Leishmania infantum co-infection in dogs. AB - A seroprevalence survey and risk analysis of Neospora caninum and Leishmania infantum was conducted in dogs from an area of the Campania region of southern Italy, in order to investigate the co-infection of these two protozoa. Blood samples were collected from 1058 asymptomatic dogs over a 18 months period. Serum samples were tested for antibodies to N. caninum and to L. infantum using the indirect fluorescent antibody test. Epidemiological data (breed, age, sex, and utilization) were collected and statistically analysed in relation to N. caninum and to L. infantum seropositivity and antibody titres. Out of the 1058 sera samples tested, 68 (6.4%) were found to have antibodies to N. caninum, and 222 (21.0%) to have antibodies to L. infantum. The co-presence of antibodies to N. caninum and to L. infantum was found in 46 (4.3%) dogs. Thus, 67.6% of the dogs positive for N. caninum also had antibodies to L. infantum. The major risk factor for N. caninum seropositivity was the presence of antibodies to L. infantum, and the major risk factor for L. infantum seropositivity was the presence of antibodies to N. caninum. In addition, high N. caninum seroprevalence was closely correlated to Boxer breed, and high L. infantum seroprevalence was correlated to masculine gender and Setter and Pit bull breeds. Low L. infantum seroprevalence was closely correlated to Yorkshire breed. The findings of this survey indicate that in the Campania region of southern Italy the co-presence of antibodies to N. caninum and to L. infantum is very common in dogs, and that infection by one protozoan seems to enhance the susceptibility to the other one. This is probably due to the immunological status of the tested dogs. PMID- 12079737 TI - Chemotherapy of canine leishmaniosis. AB - Visceral leishmaniosis is a widespread and potentially fatal disease of dogs and humans common in the Mediterranean region, the Middle East, and South America. Canine leishmaniosis is most frequently treated with the drugs meglumine antimoniate, allopurinol, amphotericin B, or a combination of meglumine antimoniate and allopurinol. Therapy with the currently used drugs often achieves temporary clinical improvement and changes in immunologic parameters with restoration of the ability to mount parasite-specific cell mediated responses and decrease in anti-leishmanial antibody titers. However, treatment usually does not prevent relapse of disease or eliminate parasite carriage. Due to the current lack of an ultimate and effective therapy for canine leishmaniosis, new drugs, delivery systems and treatment strategies are necessary to achieve a consistent parasitological cure in infected dogs. PMID- 12079738 TI - Suppressed cytotoxic T lymphocyte responses in experimental cysticercosis. AB - Mitogen-induced T cell responses are suppressed in mice infected with larvae of Taenia crassiceps. The effects of experimental infection on specific T cell responses, however, have not been examined. In the present study, we demonstrate that larval-infected mice exhibit suppressed ability to develop anti-virus specific cytotoxic T lymphocyte (CTL) responses while maintaining apparently normal natural killer (NK) cell responsiveness. PMID- 12079739 TI - Evaluation of tests for anthelmintic resistance in cyathostomes. AB - Resistance, especially to the anthelmintic benzimidazoles (BZ), has been reported in horse cyathostomes world-wide. Diagnosis of resistance has traditionally been made by faecal egg count reduction (FECR) trials, however, this technique has limitations. Some of the shortcomings may be resolved by refining the test or by using an in vitro test. FECR tests and the larval development assay (LDA) were performed on adult horses held on 15 different horse properties across a wide geographical area of NSW, Australia. FECR were measured before and 10-14 after days treatment with oxibendazole (OBZ), morantel (MOR) or ivermectin (IVM) at recommended dose rates. Eight properties were rejected following low pre treatment egg counts, leaving seven in the study. On these, the majority of larvae recovered from faecal cultures were cyathostomes. Using a definition of resistance as a FECR of <90%, resistance to OBZ was present on six properties and to MOR on two properties. Resistance to IVM was not detected. An alternative method of calculating FECR based on individual horse egg counts pre- and post treatment was developed and results from the same properties compared with the results of the LDA. For example, for the BZ, correlation coefficients of values of lethal concentration to kill 50% of population (LC50) on LDA and FECR percentages were -0.536 before and -0.704 after OBZ treatment. We conclude that the LDA has the potential to be a single visit test for detection of anthelmintic resistance in horse cyathostomes but requires further investigation and standardisation. PMID- 12079740 TI - Comparison of selective and systematic treatments to control nematode infection of the digestive tract in dairy goats. AB - Resistance to anthelmintics in nematode parasite of the digestive tract is a major concern in small ruminants and particularly in goats. One possible solution to limit the development and spread of resistance is to give treatments on a selective basis, i.e. by targeting the most susceptible animals within a flock rather than treating all the animals. In dairy goats, epidemiological studies have shown that, within a flock, goats in first lactation and the multiparous ones with the highest level of milk production were highly receptive to parasite infection. The objective of the study was to assess whether selective treatments could achieve a level of control of nematodes similar to systematic drenching. A similar experimental design was applied for two successive years. An experimental flock of 120 dairy goats was divided into two groups. All the goats from group SYS (systematic treatment) were drenched at mid-grazing season (July). In contrast, anthelmintics in group SEL (selective treatment) were restricted to the goats in first lactation plus the high producers. Overall, these treated goats represented 48% of the flock in year 1 and 66% in year 2. After treatments, both groups grazed on separate pastures. Parasitological and pathophysiological measurements were performed monthly. The results indicate a similar level of egg excretion in the two groups. The pathophysiological parameters (pepsinogen and phosphate concentrations) were also similar in the two groups, as well as the milk production for the 2 years. These results suggest that a targeted use of anthelmintics may allow efficient control of gastrointestinal nematodes whilst resulting in a predicted reduction in the selection pressure for the development of anthelmintic resistance. PMID- 12079741 TI - Myiasis in sheep in The Netherlands. AB - In 1999, among 164 randomly selected Dutch sheep farmers, a questionnaire was carried out to estimate the prevalence of myiasis in sheep and to investigate factors associated with the occurrence of myiasis. The total number of sheep and/or lambs on the reference date 1 August 1999 was 12,200: 5243 ewes, 225 rams, 3393 ewe- and 3339 ram lambs. On 86 (52.4%) of the farms 349 (2.9%) of all sheep and/or lambs contracted myiasis, of which two died. Myiasis was seen significantly more frequently on farms with over 25 ewes compared to smaller farms. Cases of myiasis were detected from April to September, with a peak (47.1%) in August. Occurrence of myiasis was most frequently associated with hot and humid weather and was mainly observed (69.1%) in the area around the tail.Ewes and ewe lambs had significantly more myiasis when compared with rams and ram lambs. There was no relationship with tail docking, with breed, with the time of shearing, with the kind of soil (clay, sand, etc.), with the environment (bush, trees, water, etc.), with the type of treatment (pour on, dipping, spraying), the used insecticides (synthetic pyrethroids, diazinon, cyromazin, etc.), the number of preventive treatments, the time of treatment or the number of observations on the herd (once a day, once a week, etc.). PMID- 12079743 TI - Polymerase chain reaction detection of Mycobacterium tuberculosis complex and Mycobacterium avium organisms in formalin-fixed tissues from culture-negative ruminants. AB - In the US eradication program for bovine tuberculosis, a definitive diagnosis depends on the isolation of Mycobacterium bovis. However, in some cases bacterial culture is unsuccessful, even though the tissue is considered suspicious by histopathology because granulomatous lesions and acid-fast organisms are present. The purpose of this study was to determine if polymerase chain reaction (PCR) tests on formalin-fixed tissue would successfully identify the organisms observed in suspect lesions from culture-negative animals. Diagnostic laboratory records were used to select paraffin blocks of tissue from 102 ruminants that had suspect microscopic lesions but no bacterial isolation. Sections from these blocks were examined with PCR primers for IS6110 to detect Mycobacterium tuberculosis complex infection, or with 16S ribosomal RNA and IS900 primers for detection of Mycobacterium avium. The PCR tests successfully identified a mycobacterial infection in 58 of 102 tissues, including 41 M. tuberculosis complex and 17 M. avium (11 subspecies paratuberculosis). These results demonstrate that PCR testing of formalin-fixed tissue, in combination with bacterial culture, may increase the effectiveness of laboratory diagnostic efforts to detect and identify the most common mycobacterial diseases of ruminants. PMID- 12079742 TI - Brucella abortus INTA2, a novel strain 19 (Delta)bp26::luc (Delta)bmp18 double mutant lacking drug resistance markers. AB - Brucella abortus INTA2, a novel mutant strain, was constructed by inactivation of two B. abortus S19 genes: bp26 and bmp18, with the objective of obtaining a mutant strain that could be compatible with a diagnostic test and have less residual virulence than strain 19. The double mutant was constructed by replacing a large section of the bp26 coding region with the luciferase (luc) coding gene, resulting in mutant strain B. abortus M1luc, followed by partial deletion of bmp18 coding sequence. Both genes were inactivated by allelic replacement assisted by sacB counter-selection. Luciferase expression was evaluated and confirmed that it is a valid marker in the construction of mutant strains. When B. abortus INTA2 was inoculated in BALB/c mice, significantly fewer colony forming units (CFUs) were recovered from mice spleens during initial phase of infection. No splenomegaly was observed in strain INTA2-immunized mice at any time suggesting that strain INTA2 has lost some residual virulence of the parental strain. Nevertheless, similar protection levels against virulent challenge were observed in mice immunized with strains INTA2 or S19. Although strain INTA2 would still induce O-side antibodies, it does not express BP26. This would allow differentiation of INTA2-vaccinated animals from animals infected with field strains by measuring anti-BP26 antibodies, either by an agglutination test or ELISA using BP26 as antigen. Altogether these results indicate that B. abortus INTA2 might be a promising vaccine strain against brucellosis. PMID- 12079744 TI - Detection and identification of salmonellas from poultry-related samples by PCR. AB - A polymerase chain reaction (PCR) assay was developed for the generic detection of Salmonella sp. and the identification of S. Enteritidis (SE), S. Gallinarum (SG), S. Pullorum (SP) and S. Typhimurium (ST) in material collected in the field from poultry. The specificity and sensitivity of the assay combined with Rappaport-Vassiliadis selective enrichment broth (PCR-RV) were determined, and field samples were analyzed to verify the validity of the method application. Specificity of the assay was tested using 29 SE, 11 SG, 10 ST and 10 SP strains, along with 75 strains of 28 other Salmonella serovars and 21 strains of other bacterial genera. The assay was 100% specific for Salmonella detection and ST identification. The primer pair for SE, SG and SP also detected S. Berta. PCR detection limits for Salmonella at the genus level were 2 ST, 8 SE, 1.1x10(3) SG and 1.8x10(5) SP cells. At the serovar level, detection limits were 7 ST, 1.2x10(3) SE, 4.4x10(7) SG and 1.8x10(6) SP cells. At the genus level, PCR-RV detected approximately 128% more positive field samples than the standard microbiological techniques and results were ready in 48h instead of 7 days. PCR RV method is diagnostic of Salmonella at the genus level and ST at the serovar level, although other tests are needed to identify SE, SG and SP to serovar level. PMID- 12079745 TI - Monoclonal antibodies specific for Campylobacter fetus lipopolysaccharides. AB - Four monoclonal antibodies (mAbs) (M1357, M1360, M1823 and M1825) which reacted with Campylobacter fetus lipopolysaccharide (LPS) core region epitopes were produced and characterized. Reactivity of these mAbs with C. fetus core LPS epitopes was determined by enzyme-linked immunosorbent assay (ELISA) with whole cell proteinase K digests and phenol-water extracted LPS, and by immunoblotting with proteinase K digests. The specificities of the four mAbs were evaluated using an indirect ELISA. One of the mAbs reacted with 42 and three of the mAbs reacted with 41 of the 42 C. fetus strains examined. No reaction was observed between the four mAbs and 32 non-C. fetus bacteria tested, with the exception of one mAb with one organism. The four mAbs reacted with serotype A and B strains indicating the presence of shared epitopes in C. fetus LPS core oligosaccharides. The specificities of three mAbs previously produced to C. fetus LPS O-antigens (M1177, M1183 and M1194) were also evaluated and no reaction was observed with these mAbs and the 32 non-C. fetus bacteria tested. Strong immunofluorescence reactions were observed with the anti-O chain mAbs and selected C. fetus strains of the homologous serotype. These anti-LPS core oligosaccharide and anti-LPS O chain mAbs are highly specific for C. fetus and are potentially useful as immunodiagnostic reagents for detection, identification and characterization of C. fetus. PMID- 12079746 TI - Quantification of bioavailable chlortetracycline in pig feces using a bacterial whole-cell biosensor. AB - Bacterial whole-cell biosensors were used to measure the concentration of chlortetracycline (CTC) in the feces of pigs. In this study, the Escherichia coli biosensor used has a detection limit of 0.03 mg/kg CTC in pig feces. The tetracycline concentration was correlated with the appearance and maintenance of fecal coliform bacteria resistant to tetracycline. Initially, large quantities of water-extractable CTC were excreted from the pigs and measurable amounts were detected even at 30 days after treatment cessation. This led to a sharp rise in the number of tetracycline resistant coliform bacteria in the feces, to within the same order of magnitude as the total coliform count. The high level of tetracycline resistance was maintained in spite of the declining concentration of tetracycline. PMID- 12079747 TI - Serological responses against the pathogenic dimorphic fungus Mucor amphibiorum in populations of platypus (Ornithorhynchus anatinus) with and without ulcerative mycotic dermatitis. AB - Mucor amphibiorum, a dimorphic fungus, causes ulcerative dermatitis and systemic infections in the platypus Ornithorhynchus anatinus in some river systems in Tasmania but apparently not in other regions of Australia. As yet there are no suitable tests for population surveys, nor for detection of internal lesions in live animals. Consequently, immunoglobulins were purified from the serum of platypuses and anti-immunoglobulin antisera were prepared in rabbits in order to develop an enzyme-linked immunosorbent assay (ELISA) for anti-M. amphibiorum antibodies. Antigens from plate-grown cultures resulted in greater signal-to noise ratios in indirect ELISA than those from broth-grown cultures. Platypuses with clinical ulcerative dermatitis had elevated anti-Mucor antibody levels compared to apparently unaffected individuals. Seroconversion was observed in one animal coincident with the development of cutaneous ulcers. The results suggested that platypuses in affected rivers were exposed to M. amphibiorum at a higher frequency than the occurrence of clinical disease. Some platypuses from New South Wales had elevated antibody levels but these increased significantly with age suggesting exposure to cross-reactive antigens, although exposure to M. amphibiorum cannot be excluded. Further studies are warranted to determine factors that result in progression from infection to disease, the occurrence of the fungus in areas where disease has not been observed and the specificity of antigen used in ELISA. PMID- 12079749 TI - A comparative study of the intestinal microbiota of healthy horses and those suffering from equine grass sickness. AB - This study compares quantitatively the microbiota of the gastrointestinal tract of healthy horses with that of horses with equine grass sickness (EGS). Faecal and ileal samples were cultured quantitatively on selective and non-selective media. Confirmed anaerobes were identified to species level. Overall faecal counts gave a ratio of aerobes:anaerobes of approximately 1:1. However, the mean counts in healthy horses of 4.4x10(8) aerobes:3.7x10(8) anaerobes per gram wet weight were different from counts in EGS (means were 10-100-fold higher), with statistically significant differences for the anaerobes (p=0.04). There were 10 100-fold more anaerobic cocci in EGS samples compared to healthy controls. Most of the seven species of anaerobic cocci were found in both healthy horses and EGS. Differences in clostridia isolated between health and disease were notable: fourteen species were isolated from EGS cases, compared to only one (C. bifermentans) in controls. The mean faecal clostridial counts in chronic disease were higher than in controls (10-fold) and in acute EGS (100-fold). In contrast, mean counts for ileal samples from acute cases, showed a 10-fold increase for clostridia compared to 1000-fold reduction in chronic cases (compared to faecal counts). Results indicate an increase in the bacterial numbers in the GI tract of animals with EGS compared to the controls and clostridia are prominent in EGS. Whether the increase in clostridia is the cause of GI stasis or a consequence remains uncertain. PMID- 12079748 TI - Pulsed field gel electrophoresis for animal Salmonella enterica serovar Typhimurium isolates in Taiwan. AB - Salmonella enterica subspecies enterica serovar Typhimurium is a common pathogen for humans and animals. In order to trace the clonal relationship and to find the circulating strains between human and animal isolates, chromosomal DNAs from 87 serovar Typhimurium strains isolated from animals (pigs were the majority) were subjected to XbaI and SpeI digestion and pulsed field gel electrophoresis (PFGE). For the 87 animal isolates, 38 PFGE pattern combinations were obtained. As the subtyping results from animal isolates were compared with those from the 45 human isolates, it was found that 14 of the animal isolates and 13 of the human isolates shared a common PFGE pattern combination, i.e., pattern XgSf (or called X5S4). When these human and animal isolates were subjected to antibiotic susceptibility test using 11 antibiotics, it was found that strains of pattern XgSf (X5S4) belong to a common antibiogram pattern which is tetracycline, gentamicin, ampicillin, streptomycin and chloramphenicol resistant. Since most of the animal and human strains in pattern XgSf were originally isolated from various areas over different years, strains of this PFGE pattern may be the most epidemic strains which circulating between human and animal sources. PMID- 12079751 TI - Tau neurotoxicity without the lesions: a fly challenges a tangled web. AB - Models of neurodegenerative disorders are challenging the classical defining role of tangles in neurotoxicity. In flies, tau overexpression is sufficient to cause neuronal death without the formation of fibrillar aggregates. This parallels observations in models of polyglutamine disorders and suggests that aggregated protein might not be the toxic species responsible for neuronal dysfunction and cell death. PMID- 12079752 TI - RIM1: an edge for presynaptic plasticity. AB - Pioneering work suggests that a synaptic active zone protein, RIM1, regulates both short- and long-term glutamatergic presynaptic plasticity at certain synapses. In short-term plasticity, RIM1 accelerates the priming of synaptic vesicles for fusion; by contrast, in long-term potentiation of mossy fiber synapses in the hippocampal CA3 region, phosphorylated RIM1 acts through an unknown molecular pathway to enhance release of the excitatory neurotransmitter glutamate. PMID- 12079753 TI - Ephs and ephrins close ranks. AB - The past decade has seen remarkable advances in identification of the proteins regulating axon guidance and synapse formation. Understanding the structural and molecular basis of their signaling properties is now the task at hand. The recently characterized crystal structure of the complex formed between the ligand binding domain of EphB2 and the ectodomain of its binding partner ephrin-B2 provides an insight into the recognition and signal transduction mechanisms of this large multifunctional family of surface receptors. This heterotetrameric complex reveals a cyclic arrangement of subunits not previously seen in any receptor-ligand structure, and provides a basis for class specificity of binding. PMID- 12079754 TI - Sex, cells and signals in the developing brain. PMID- 12079755 TI - Knockout mice: simple solutions to the problems of genetic background and flanking genes. AB - Inducing null mutations by means of homologous recombination provides a powerful technique to investigate gene function and has found wide application in many different fields. However, it was realized some time ago that the specific way in which such knockout mutants are generated can be confounding, making it impossible to separate the effects of the induced null mutation from those of alleles originating from the embryonic stem cell donor. In addition, effects from null mutations can be altered on different genetic backgrounds. Here we present some simple breeding strategies to test for flanking gene effects that are compatible with the recommendations of the Banbury Conference on Genetic Background in Mice and with common practices of creating and maintaining mouse knockout lines. PMID- 12079756 TI - Rodent models of prefrontal cortical function. AB - In this article, we consider whether studies in rats can provide useful information regarding the debate about the functions of the primate prefrontal cortex. At a superficial level, comparison of regional specializations within the prefrontal cortices of different species suggests functional correspondence. Unfortunately, the nature of functional specialization in primate prefrontal cortex is controversial, and data supporting the idea of homology between specific areas of rat and primate prefrontal cortex are weak. Nevertheless, we argue here that studies of the computational functions within the relatively undifferentiated prefrontal cortex of rats can shed light on processing in primate prefrontal cortex. PMID- 12079762 TI - The planum temporale as a computational hub. AB - It is increasingly recognized that the human planum temporale is not a dedicated language processor, but is in fact engaged in the analysis of many types of complex sound. We propose a model of the human planum temporale as a computational engine for the segregation and matching of spectrotemporal patterns. The model is based on segregating the components of the acoustic world and matching these components with learned spectrotemporal representations. Spectrotemporal information derived from such a 'computational hub' would be gated to higher-order cortical areas for further processing, leading to object recognition and the perception of auditory space. We review the evidence for the model and specific predictions that follow from it. PMID- 12079763 TI - Has electrical growth cone guidance found its potential? AB - Many neurobiologists spurn the existence and use of direct-current (DC) electric fields (EFs) in nervous system development and regeneration. This is despite direct measurement of EFs in embryos and adults, and evidence that EFs are required for normal development, dramatically influence the rate and direction of nerve growth in vitro, and promote nerve regeneration in vivo. The notion that growth cones use EFs as guidance cues was dismissed partly because there was no convincing evidence that naturally occurring EFs influence nerve growth at the single-cell level in vivo. Recent work indicates that growth cones can be guided by EFs in vivo and, intriguingly, that in vitro guidance by chemotropic gradients and EFs might invoke similar mechanisms. Ongoing clinical trials to assess the effectiveness of DC EFs in promoting the regeneration of human spinal cord could allow EFs to achieve their potential. PMID- 12079764 TI - Does glutamate image your thoughts? AB - Functional imaging methods exploit the relationship between neuronal activity, energy demand and cerebral blood flow to functionally map the brain. Despite the increasing use of these imaging tools in basic and clinical neuroscience, the neurobiological processes underlying the imaging signals remain unclear. Recently, interest has been focused on uncovering the signals that trigger the metabolic and vascular changes accompanying variations in neuronal activity. Advances in this field have demonstrated that release of the major excitatory neurotransmitter glutamate initiates diverse signaling processes between neurons and astrocytes, and that this signaling could be crucial for the occurrence of brain imaging signals. In this article we review the hypothesis that glutamate represents a common trigger for both neurometabolic and neurovascular coupling. PMID- 12079765 TI - Interactive nervous system development: control of cell survival in Drosophila. AB - The non-autonomous control of cell survival has long been thought to be a mechanism of adjusting cell populations in the vertebrate nervous system, enabling connectivity and myelination to produce a functional brain. Despite cellular evidence that analogous mechanisms occur in invertebrates, scepticism has long reigned over whether they operate in model organisms such as Drosophila. This has led to speculation that there are inherent differences between the development and evolution of simple brains and the brains of vertebrates. The great paradox has, until recently, been the absence of molecular evidence of trophic factors in Drosophila. Recent data have finally shown that EGFR (epidermal-growth-factor receptor) ligands function in the Drosophila CNS to maintain glial survival. Trophic interactions are, thus, a general mechanism of nervous system development. PMID- 12079766 TI - The special nature of human walking and its neural control. AB - Walking the way we do is inherently unstable. Sophisticated neurological control systems are required to ensure that we progress and maintain our balance at the same time. Most of what is known about the functional organization of these neurological control systems is inferred from studies on animals. Here, I compare selected studies on the neural control of human walking with similar studies in reduced animal preparations. The simple monosynaptic reflex appears to be controlled by comparable mechanisms in walking cats and humans. However, peripheral feedback mechanisms suggested to contribute to the switch from stance to swing on the basis of experiments in reduced cat preparations have little influence during human walking. A cat whose spinal cord has been completely transected can be made to walk on a treadmill by drug injections, but such an immediate effect of pharmacological intervention is not seen in humans. However, there have been reports that pharmacological intervention can improve the walking of patients with incomplete spinal cord injury, especially when pharmacological treatment is combined with training. PMID- 12079769 TI - European research on the functional effects of dietary antioxidants - EUROFEDA. PMID- 12079770 TI - Bioavailability and metabolism. PMID- 12079771 TI - Biomarkers. PMID- 12079772 TI - Antioxidants, reactive oxygen and nitrogen species, gene induction and mitochondrial function. PMID- 12079774 TI - Archemix. Nucleic acid platforms. PMID- 12079773 TI - European Research on the Functional Effects of Dietary Antioxidants (EUROFEDA). Conclusions. PMID- 12079776 TI - Dissecting histone deacetylase function. AB - Recent research describes the use of chromatin immunoprecipitation and intergenic chromosomal-DNA microarrays to analyze HDAC function genome-wide. The next step in realizing the full potential of these analyses will be to develop specific and temporal control over HDAC perturbation. PMID- 12079775 TI - Building a metal binding domain, one half at a time. AB - The recently determined structure of a zinc binding peptide reveals that a particular sequence can adopt one stable fold as an isolated peptide but adopt an alternative structure as part of a larger protein domain. PMID- 12079777 TI - Recognizing a something when your library sees it. AB - Advances are needed in random-display technologies to more tightly link drug actions and functions to the genes that control physiological processes. The reports discussed here explore two sides of these issues-generating new library formats and identifying the targets of drug ligands. PMID- 12079778 TI - Metallochaperones: bind and deliver. AB - Metallochaperones deliver metal ions directly to target proteins via specific protein-protein interactions. Recent research has led to a molecular picture of how some metallochaperones bind metal ions, recognize their partner proteins, and accomplish metal ion transfer. PMID- 12079780 TI - Drug receptor identification from multiple tissues using cellular-derived mRNA display libraries. AB - The use of display technologies to identify small molecule receptors from proteome libraries would provide a significant advantage in drug discovery. We have used mRNA display to select, based on affinity, proteins that bind to a drug of interest. A library of mRNA-protein fusion molecules was constructed from human liver, kidney, and bone marrow transcripts and selected using an immobilized FK506-biotin conjugate. Three rounds of selection produced full length FKBP12 (FK506 binding protein 12 kDa) as the dominant clone. An analogous method was also used to map the minimal drug binding domain within FKBP12. Using this approach, it is anticipated that mRNA display could eventually play a key role in the discovery and characterization of new drug receptor interactions. PMID- 12079779 TI - Path of an RNA ligand around the surface of the vaccinia VP39 subunit of its cognate VP39-VP55 protein heterodimer. AB - VP39 is a vaccinia virus-encoded RNA modifying protein with roles in the modification of both mRNA ends. At the 3' end it acts as a processivity factor for the vaccinia poly(A) polymerase (VP55), promoting poly(A) tail elongation. Despite VP39's three-dimensional structure having been elucidated along with details of its mode of mRNA 5' end binding, the VP39-VP55 heterodimer's molecular mechanism of processivity is largely unknown. Here, the area immediately above almost the entire surface of the VP39 subunit was probed using chemical reporters, and the path of a previously unidentified RNA binding site was revealed. The path was indicated to fall within a cleft formed by the intersubunit interface and was consistent with both a previously reported model of the heterodimer-nucleic acid ternary complex and the known function of the heterodimer in processive poly(A) tail elongation. PMID- 12079781 TI - Investigation of RNA-protein and RNA-metal ion interactions by electron paramagnetic resonance spectroscopy. The HIV TAR-Tat motif. AB - Electron paramagnetic resonance (EPR) spectroscopy was used to investigate changes in dynamics of spin-labeled nucleotides in the TAR RNA (U23, U25, U38, and U40) upon binding to cations, argininamide, and two peptides derived from the Tat protein. Nearly identical changes in dynamics were obtained for either calcium or sodium ions, indicating the absence of a calcium-specific structural change for the TAR RNA in solution that had previously been suggested by crystallographic data. Similar dynamic signatures were obtained for two Tat derived peptides that have the same important binding determinant (R52) and similar binding affinities to the TAR RNA. However, U23 and U38 were substantially less mobile for the wild-type peptide (YGRKKRRQRRR) than for the mutant (YKKKKRKKKKA), demonstrating that, flanking R52, amino acids in the wild type sequence make specific contacts to the RNA. PMID- 12079782 TI - Structure of TAR RNA complexed with a Tat-TAR interaction nanomolar inhibitor that was identified by computational screening. AB - HIV-1 TAR RNA functions critically in viral replication by binding the transactivating regulatory protein Tat. We recently identified several compounds that experimentally inhibit the Tat-TAR interaction completely at a 100 nM concentration. We used computational screening of the 181,000-compound Available Chemicals Directory against the three-dimensional structure of TAR [1]. Here we report the NMR-derived structure of TAR complexed with acetylpromazine. This structure represents a new class of compounds with good bioavailability and low toxicity that bind with high affinity to TAR. NMR data unambiguously show that acetylpromazine binds only to the unique 5' bulge site to which the Tat protein binds. Specificity and affinity of binding are conferred primarily by a network of base stacking and hydrophobic interactions. Acetylpromazine alters the structure of free TAR less than Tat peptides and neomycin do. PMID- 12079783 TI - Saccharide display on microtiter plates. AB - New insight into the importance of carbohydrates in biological systems underscores the need for rapid synthetic and screening procedures for them. Development of an organic synthesis-compatible linker that would attach saccharides to microtiter plates was therefore undertaken to facilitate research in glycobiology. Galactosyllipids containing small, hydrophobic groups at the anomeric position were screened for noncovalent binding to microtiter plates. When the lipid component was a saturated hydrocarbon between 13 and 15 carbons in length, the monosaccharide showed complete retention after aqueous washing and could be utilized in biological assays. This alkyl chain was also successfully employed with more complex oligosaccharides in biological assays. In light of these findings, this method of attachment of oligosaccharides to microtiter plates should be highly efficacious to high-throughput synthesis and analyses of carbohydrates in biological assays. PMID- 12079784 TI - Engineering deoxysugar biosynthetic pathways from antibiotic-producing microorganisms. A tool to produce novel glycosylated bioactive compounds. AB - A plasmid (pLN2) was generated in which genes involved in the biosynthesis of L oleandrose in the oleandomycin producer Streptomyces antibioticus ATCC11891 were cloned. pLN2 was used to direct the biosynthesis of different deoxysugars by exchanging and/or adding genes from other antibiotic biosynthetic clusters. Transfer of the synthesized deoxysugars to the tetracenomycin C aglycon, 8 demethyl-tetracenomycin C, through the use of the "sugar flexible" glycosyltransferase ElmGT, validated the system. Several pLN2 derivatives were constructed by replacement of the oleU 4-ketoreductase gene by different 4 ketoreductase genes. Some of them, such as EryBIV and UrdR, reduced the keto group of the 4-keto intermediates, generating L-olivosyl and D-olivosyl derivatives, respectively. The system was also used to generate an L-rhamnosyl derivative (through a two-gene deletion) and an L-rhodinosyl derivative (through a combination of a gene replacement and a gene addition). PMID- 12079785 TI - Redox-active cyclic bis(cysteinyl)peptides as catalysts for in vitro oxidative protein folding. AB - The active-site hexapeptides of glutaredoxin (Grx), thioredoxin (Trx), protein disulfide isomerase (PDI), and thioredoxin-reductase (Trr) containing the common motif Cys-Xaa-Yaa-Cys were conformationally restricted by backbone cyclization, and their redox potentials were found to increase in the rank order of Trr < Grx < Trx < PDI peptide, with E'(0) values ranging between -204 mV and -130 mV. In each peptide the thiol pK(a) of one Cys residue was found to be lower than the other (e.g., 7.3 against 9.6 in the PDI peptide). Both the yield and rate of refolding of reduced RNase A in the presence of the bis(cysteinyl)peptides increased with the oxidizing character of the cyclic compounds. These results show that small peptides can function as adjuvants for the in vitro oxidative folding of proteins. PMID- 12079786 TI - A bifunctional tRNA for in vitro selection. AB - In vitro selection is a powerful approach for generating novel aptamers and catalysts. Currently, several methods are being developed to extend this technique to proteins. In principle, selection methods could be applied to any library whose members can be replicated. Here, we describe a bifunctional tRNA that fuses translation products to their mRNAs. The utility of peptide-tRNA-mRNA fusions for in vitro selection was illustrated by the selective enrichment of tagged peptides-together with their mRNAs-by affinity chromatography. Our system can generate libraries larger than 10(11). Because library members can be copied and amplified, they provide a means for applying in vitro selection procedures to peptides and proteins. Furthermore, because the system is amenable to translation with misacylated tRNAs, a wide range of unusual monomers could be used to make libraries of nonstandard polymers for selection experiments. PMID- 12079787 TI - Crystal structure of a complex between the aminoglycoside tobramycin and an oligonucleotide containing the ribosomal decoding a site. AB - Aminoglycoside antibiotics target the decoding aminoacyl site (A site) on the 16S ribosomal RNA and induce miscoding during translation. Here, we present the crystal structure, at 2.54 A resolution, of an RNA oligonucleotide containing the A site sequence complexed to the 4,6-disubstituted 2-deoxystreptamine aminoglycoside tobramycin. The three aminosugar rings making up tobramycin interact with the deep-groove atoms directly or via water molecules and stabilize a fully bulged-out conformation of adenines A(1492) and A(1493). The comparison between this structure and the one previously solved in the presence of paromomycin confirms the importance of the functional groups on the common neamine part of these two antibiotics for binding to RNA. Furthermore, the analysis of the present structure provides a molecular explanation to some of the resistance mechanisms that have spread among bacteria and rendered aminoglycoside antibiotics inefficient. PMID- 12079788 TI - Assembly of p-selectin ligands on a polymeric template. AB - High-affinity receptor-ligand interactions frequently involve molecular interactions at two distinct sites. A derivatized polyacrylic-based polymer was synthesized to allow substitution with multiple ligands (e.g., L(1) and L(2)) on the backbone. Two-site P-selectin-ligand interactions were first studied with SiaLe(x) (L(1)) and tyrosine sulfate (L(2)) covalently incorporated onto the flexible polymer. In competition assays, a marked synergistic inhibitory effect was observed when the polymer presented both L(1) and L(2) as opposed to either ligand alone. In a second approach, the SiaLe(X) ligand was reduced in complexity so that L(1) was fixed as Le(x) or Le(a), and alternative L(2) groups (to mimic sialic acid) were investigated. Certain combinations of L(1) and L(2) were better antagonists of P-selectin than SiaLe(x) itself. These approaches offer the potential of facilitating the discovery of novel inhibitors of receptors or enzymes. PMID- 12079789 TI - Ocular wave-front aberration statistics in a normal young population. AB - Monochromatic ocular aberrations in 108 eyes of a normal young population (n=59) were studied. The wave-front aberration were obtained under natural conditions using a near-infrared Shack-Hartmann wave-front sensor. For this population and a 5 mm pupil, more than 99% of the root-mean square wave-front error is contained in the first four orders of a Zernike expansion and about 91% corresponds only to the second order. Comparison of wave-fronts aberrations from right and left eye in 35 subjects, showed a good correlation between most of the second- and third order terms and a slight (but not clear) tendency for mirror symmetry between eyes. PMID- 12079790 TI - Optic flow and scene structure do not always contribute to the control of human walking. AB - Using displacing prisms to dissociate the influence of optic flow and egocentric direction, previous research (Current Biology 8 (1998) 1191) showed that people primarily use egocentric direction to control their locomotion on foot, rather than optic flow. When wearing displacing prisms, participants followed the curved path predicted by the use of simple egocentric direction, rather than a straight path, as predicted by the use of optic flow. It has previously been suggested that, in rich visual environments, other visual information including optic flow and static scene structure may influence locomotion in addition to direction. Here we report a study where neither scene structure nor optic flow have any influence on the control of walking. Participants wearing displacing prisms walked along a well-lit corridor (containing rich scene structure and flow) and along the same corridor in darkness (no scene structure or flow). Heading errors were not significantly different between the dark and light conditions. Thus, even under conditions of rich scene structure and high flow when walking in a well-lit corridor, participants follow the same curved paths as when these cues are not available. These results demonstrate that there are conditions under which visual direction is the only useful source of visual information for the control of locomotion. PMID- 12079791 TI - Color appearance of spatial pattern: the role of increments and decrements. AB - In a number of recent adaptational studies evidence for a different processing of incremental and decremental cone signals has been reported. The present study examined whether such asymmetries occur in spatial pattern as well. Subjects set color matches between a uniform, 2 degrees matching box and bars within squarewave patterns. The squarewaves varied in spatial frequency, color direction, and contrast. For all three cone signals the asymmetric matches showed clear evidence for increment-decrement asymmetries: Although both incremental and decremental matches scaled roughly linearly with pattern contrast, in general, the scalings for the two types of color signals differed. This difference in scaling increased with spatial frequency, thus leading to an increase in the size of the increment-decrement asymmetry with spatial frequency. The matches were well described by means of two-stage models, consisting of a color transformation in the first stage and a pattern-dependent scaling in the second stage. Analyses based on these pattern-color separable models suggest that the asymmetries are mediated mainly through a white-black mechanism and much less, if at all, through a red-green and yellow-blue mechanism. PMID- 12079792 TI - Infant photometry: are mean adult isoluminance values a sufficient approximation to individual infant values? AB - Individual differences in isoluminance values were studied in infants and adults using a motion nulling paradigm. Two luminance-modulated sinusoidal grating components (spatial frequency=0.25 cpd, temporal frequency=5.6 Hz, speed=22.4 deg/s) were superimposed and moved in opposite directions across a color video screen. The contrasts of the two components were traded off to determine motion nulls. Two conditions were used: red/black vs. green/black, and red/black vs. blue/black grating components. An eye movement based response measure was used for infant subjects, and an average of 308 trials per infant were obtained. As observed in earlier studies, the mean motion null values for infants and adults were highly similar in each condition. The standard errors of motion null values for individual subjects were very small. Individual differences among infants were also small, and were clearly measurable only in the red/black vs. blue/black condition. The close similarity of mean null values, combined with the small individual differences among infants, supports the idea that under the right circumstances mean adult isoluminance values can be used as a sufficient approximation to individual infant isoluminance values in studies of infant color vision. These circumstances are discussed and evaluated in detail. PMID- 12079793 TI - Time-dependent changes of rod influence on hue perception. AB - Hue-naming was used in conjunction with a probe-flash procedure to determine the time-course of rod-mediated effects on hue appearance across the spectrum. Two types of rod influence on hue are distinguishable on the basis of differences in both spectral specificity and time course of effect: (1) a "faster" rod influence enhances green relative to red and (2) a "slower" rod influence enhances short wavelength red relative to green and blue relative to yellow. The results show that there are separable rod hue biases that operate over different time courses and that the overall rod influence on hue appearance depends importantly on the temporal properties of the stimuli, presumably because rods interact in different ways with different portions of the neural pathways that mediate human color vision. PMID- 12079794 TI - Fixational eye movements are not affected by abrupt onsets that capture attention. AB - Recent work shows that abrupt onsets reflexively capture attention and trigger saccades that compete with voluntary saccades. To test whether oculomotor capture occurs when no saccade is being planned, we measured fixational eye movements in the absence or presence of an abrupt onset at peripheral locations. We found no effect of abrupt onset location on the average pattern of eye movements during fixation. We conclude that the capture of eye movements by an abrupt onset only happens when the oculomotor system has been preset to make a saccade. This implies that the oculomotor system is not obligatorily driven by events in the visual array. PMID- 12079795 TI - Can spatio-temporal energy models of motion predict feature motion? AB - Current "spatio-temporal energy" models of how we perceive pattern motion have been very successful in helping us to understand the mechanisms of motion perception. Although they have been supported by a large number of physiological and psychological studies, they have so far not provided a complete explanation for a number of results. These results emerge from experiments concerned with predicting perceived motion direction from patterns comprising two or more components. It has been suggested that these results are more consistent with an earlier type of model based on the motion of two-dimensional features. This paper briefly describes how three generic spatio-temporal energy models have been extended to predict motion derived from two-component stimuli. A new model is then presented that utilises similar architecture to the two-stage spatial temporal energy model proposed by Adelson and Movshon (Nature 300 (1982) 523). The first stage is a spatial temporal filtering stage and the second stage computes the intersection of constraints (IOC), an important constraint used in combining motion information across two or more components. In the model presented here the second stage is different. A directional spatial second derivative is used to extract zero-crossings at the component level, i.e. gratings. If any zero-crossing falls in the same spatial position for two or more components its displacement is tracked using a nearest neighbour match. Tracking these 'intersecting zero-crossings' essentially computes the IOC but also provides other properties that predict non-IOC motion, and second-order component motion. Surprising new insights are described into how current spatio-temporal energy models may also account for these results. However, unlike the model presented here, they rely on operations carried out on the two-dimensional pattern. PMID- 12079796 TI - Age-related changes in refractive index distribution and power of the human lens as measured by magnetic resonance micro-imaging in vitro. AB - We report a new technique for non-invasively mapping the refractive index distribution through the eye lens using magnetic resonance micro-imaging. The technique is applied to map the refractive index distribution throughout the sagittal plane of 18 human eye lenses ranging in age from 14 to 82 years in vitro. The results are compared with standard models for the human eye lens. They confirm that the refractive index distribution, when plotted as a function of normalised lens radius, is a function of lens age and differs both between the equatorial and axial directions and between the anterior and posterior halves of the optical axis. The refractive index of the lens nucleus exhibits a significant reduction with age amounting to 3.4+/-0.6 x 10(-4) years(-1). The contribution of the gradient index (GRIN) to the lens power decreases by 0.286+/-0.067 D/year, accounting almost entirely for the estimated overall change in lens power with age for these lenses, which were probably in their most accommodated state. The results provide experimental verification of hypothesised changes in the GRIN that have previously been invoked as contributing to presbyopia and support the hypothesis that changes in the GRIN are sufficient to offset effects of increasing curvature of human lenses with age in their unaccommodated state. PMID- 12079797 TI - Has lower-dose HRT come of age? PMID- 12079799 TI - Help for the other incontinence. PMID- 12079798 TI - Nutrition. Fish for good health. PMID- 12079800 TI - Wound healing. The anatomy of healing and scarring. PMID- 12079801 TI - By the way doctor. I've heard a lot about what I should do during pregnancy to have a healthy baby, but I'd like to know what I can do now, before I become pregnant. PMID- 12079803 TI - Lest we forget. II: Alzheimer's disease. PMID- 12079802 TI - By the way doctor. If you eat a food that you're allergic to, does it have to go all the way through the digestive tract to have an allergic effect? Wouldn't that make several meals suspect? PMID- 12079804 TI - Male infertility. PMID- 12079805 TI - Prostate cancer: three research advances. PMID- 12079807 TI - Hypnosis: theory and application. Part II. PMID- 12079806 TI - On call. I am a 64-year-old man, and I've always been proud of my perfect health record. I've also been proud of my full head of hair, even after the gray started creeping in. Four months ago I caught pneumonia and spent eight days in the hospital (three in intensive care). It took a while, but I'm finally back to normal - except that my hair is falling out. It comes out in clumps when I shampoo or even comb it, and it's gotten noticeably thin all over. I remember reading about Propecia in your newsletter but I don't have the old issue. Should I try the medication? PMID- 12079808 TI - Helping children affected by trauma and death. PMID- 12079810 TI - Growing older, feeling better. PMID- 12079809 TI - Managing and averting anger. PMID- 12079811 TI - Placebos in the brain. PMID- 12079812 TI - Eating disorders and culture. PMID- 12079813 TI - Auto safety. Is it time to get off the road? PMID- 12079814 TI - Women's health. Mammography: why is there a debate? PMID- 12079815 TI - Food and drink. Red wine headaches. PMID- 12079816 TI - Hot weather and health. The health benefits of air conditioning. PMID- 12079817 TI - By the way, doctor. I read that they're now selling a cheaper version of Mevacor. But I've also heard that Mevacor isn't as effective as the other statins. What do you think? PMID- 12079818 TI - By the way, doctor. I read in the paper that cholesterol might be good for the brain and prevent Alzheimer's disease. So I'm wondering if taking a statin is really such a good idea. PMID- 12079819 TI - By the way, doctor. I use aspirin very rarely--maybe half a dozen tablets a year. I've found that even when I get the smallest bottle of aspirin, the expiration date passes with most of the contents unused. I went looking in local stores, and the latest expiration date I could find on any bottle was October 2003. Does aspirin really become unsafe or ineffective that soon? PMID- 12079820 TI - Off-pump bypass. PMID- 12079821 TI - Coffee and the heart. PMID- 12079822 TI - Wider use for implantable defibrillators. PMID- 12079823 TI - No holiday for hard-working statins. PMID- 12079824 TI - A pinch of niacin raises HDL. PMID- 12079825 TI - Low heart risk after radiation therapy for breast cancer. PMID- 12079826 TI - Ask the doctor. My total cholesterol level is well below 200. My LDL is low, my HDL high, and my triglycerides and homocysteine are both in the "desirable" range. The one thing out of whack is that my lipoprotein(a) level is more than double the "normal" limit. My cardiologist wants me to take niacin to bring it down. I can't find any information on how serious a problem this is, especially in the context of otherwise good cholesterol levels and a healthy lifestyle. (I exercise 5 days a week; eat a low-fat, high-fiber diet; and am not overweight.). PMID- 12079827 TI - Ask the doctor. I am a 70-year-old man with high blood pressure that I control with medication, diet, and exercise. My mother, aunt, and maternal grandfather all died in their 50s after a single stroke. Is there an inherited tendency to hemorrhagic stroke? Are there any precautions I can take? PMID- 12079829 TI - Identification of susceptibility genes for complex metabolic diseases. AB - There are few successful attempts to identify genes for common, non-Mendelian diseases such as diabetes, hyperlipidemia, hypertension, etc. Such common disorders are typically both metabolically and genetically complex and the genetic technologies to identify their underlying susceptibility genes are still in their infancy. Nonetheless, genetic strategies have emerged that, when the technologies are fully developed, should allow similar success rates as for Mendelian diseases. PMID- 12079830 TI - PPARs: transcription factors controlling lipid and lipoprotein metabolism. AB - Nuclear receptors are transcription factors that are activated by ligands and subsequently bind to regulatory regions in target genes, thereby modulating their expression. Nuclear receptors thus allow the organism to integrate signals coming from the environment and to adapt by modifying the expression levels of relevant genes. The peroxisome proliferator-activated receptors (PPARs) alpha, beta/delta, and gamma constitute a subfamily of nuclear receptors. PPARalpha has been shown to bind and to be activated by leukotriene B4 and the hypolipidemic drugs of the fibrate class; PPARbeta/delta ligands are polyunsaturated fatty acids and prostaglandins; while prostaglandin J2 derivatives and the antidiabetic glitazones are, respectively, natural and synthetic ligands for PPARgamma. Upon binding and activation by their ligands, they regulate the transcription of numerous genes involved in intracellular lipid metabolism, lipoprotein metabolism, and reverse cholesterol transport in a subtype- and tissue-specific manner. PPARs therefore constitute interesting targets for the development of therapeutic compounds useful in the treatment of disorders of lipid and lipoprotein metabolism. PMID- 12079831 TI - SREBP-1: gene regulatory key to syndrome X? AB - Combined appearance of different cardiovascular risk factors seems to be more prevalent in individuals with decreased insulin sensitivity and increased visceral obesity, thereby being components of the so-called metabolic syndrome or syndrome X. Alterations in the abundance and activity of transcription factors lead to complex dysregulation of gene expression, which might be a key to understand insulin resistance-associated clinical clustering of coronary risk factors at the cellular or gene regulatory level. Recent examples are members of the nuclear hormone receptor superfamily-for example, peroxisome proliferator activated receptors (PPARs) and sterol regulatory element-binding proteins (SREBPs). Besides their regulation by metabolites and nutrients, these transcription factors are also targets of hormones (like insulin and leptin), growth factors, inflammatory signals, and drugs. Major signaling pathways coupling transcription factors to extracellular stimuli are the MAP kinase cascades. We have recently shown that SREBPs appear to be substrates of MAP kinases and propose that SREBP-1 might play a role in the development of cellular features belonging to lipid toxicity and possibly syndrome X. Thus, the metabolic syndrome appears to be not only a disease or state of altered glucose tolerance, plasma lipid levels, blood pressure, and body fat distribution, but rather a complex clinical phenomenon of dysregulated gene expression. PMID- 12079832 TI - PPARgamma, an X-ceptor for Xs. AB - Evidence from both human genetic studies and characterization of peroxisome proliferator-activated receptor gamma (PPARgamma) knockout mice suggested that the prime function of PPARgamma is fat formation and that its role in insulin sensitization might be secondary to this function. The thrifty function of PPARgamma was most likely evolutionary beneficial, but might in "times of plenty" contribute to the pathogenesis of disorders, such as obesity, insulin resistance, type 2 diabetes, and hyperlipidemia, often commonly referred to as "syndrome X". This role of PPARgamma in these diseases also questions the eventual therapeutic benefits of pure PPARgamma activation, which is associated with an increase in adipose tissue mass. We characterized a new chemical class of PPARgamma agonists, that is, FMOC-l-leucine (FLL). FLL induces a different conformation of PPARgamma relative to classical PPARgamma ligands. Mass spectrometry indicates that two molecules of FLL bind to a single PPARgamma molecule, making its mode of receptor interaction distinctive. FLL recruits a different set of coactivators and activates PPARgamma with a lower potency, but a similar maximal efficacy, relative to known PPARgamma ligands. In contrast, FLL is a more effective insulin sensitizer than current PPARgamma agonists, an effect potentially linked to its weak adipogenic activity. These data make a strong point for potential therapeutic benefits of PPARgamma modulation rather than activation. PMID- 12079833 TI - Sterol regulatory element-binding proteins (SREBPs) as regulators of lipid metabolism: polyunsaturated fatty acids oppose cholesterol-mediated induction of SREBP-1 maturation. AB - Cellular cholesterol and fatty acid metabolism in mammals is controlled by a family of transcription factors called sterol regulatory element-binding protein isoforms, three of which (SREBP-1a, 1c, and 2) are well characterized. These proteins, which are synthesized as precursors, are inserted into the endoplasmic reticulum (ER) membrane with both the amino and carboxylic acid domains facing the cytosolic face of the membrane. In sterol-deficient cells, proteolytic cleavage of SREBPs occurs, thereby releasing their N-terminal mature and active forms and enabling them to enter the nucleus, where they bind to the sterol regulatory response element (SRE) and/or E-box sequences and activate genes involved in cholesterol, triglyceride, and fatty acid biosynthesis. Of the three SREBP isoforms, SREBP-1c gene expression is induced by cholesterol and repressed by polyunsaturated fatty acids (PUFA). We have examined the changes in SREBP-1c mRNA and protein levels as well as the mRNA levels of several SREBP-1c target genes when a high-cholesterol diet is combined with diets rich in PUFA of the n-6 series. Our studies show that PUFA oppose the cholesterol-mediated SREBP-1 maturation without affecting the cholesterol-mediated increase of SREBP-1c mRNA and precursor protein. The decrease in SREBP-1 mature protein paralleled the decrease in mRNAs for genes of fatty acid and cholesterol biosynthesis, such as HMG-CoA synthase and fatty acid synthase, but interestingly gene expression of stearoyl-CoA desaturase 1 (SCD1) was instead induced. These studies suggest that the main point of control of PUFA-mediated suppression of lipogenic gene expression is the inhibition of SREBP-1 maturation. The studies also reveal that the induction of SCD1 gene expression by cholesterol occurs through a mechanism independent of SREBP-1 maturation. PMID- 12079834 TI - Hyperglycemia and insulin resistance: possible mechanisms. AB - Sustained hyperglycemia impairs insulin-stimulated glucose utilization and glycogen synthesis in human and rat skeletal muscles, a phenomenon referred to clinically as glucose toxicity. In rat extensor digitorum longus (EDL) muscle preparations preincubated for 2-4 h in a hyperglycemic medium (25 mM vs. 0 mM glucose), we have shown that the ability of insulin to stimulate glucose incorporation into glycogen is impaired. Interestingly, this was associated with a decreased activation of Akt/PKB, but not its upstream regulator, PI3-kinase. A similar pattern of signaling abnormalities has been observed in adipocytes, L6 muscle cells, C2C12 cells, and (as reported here) EDL incubated with C(2) ceramide. On the other hand, no increase was observed in ceramide mass in EDL incubated with 25 mM glucose. Hyperglycemia-induced insulin resistance also has been described in adipocytes, where it has been linked to activation of novel and conventional protein kinase C isoforms that phosphorylate the insulin receptor and IRS. In addition, we have recently shown that hyperglycemia causes insulin resistance in cultured human umbilical vein endothelial cells (HUVEC). Here, it was associated with an increased propensity to apoptosis and, as in muscle, with an impaired ability of insulin to activate Akt. Interestingly, these effects of hyperglycemia and an increase in diacylglycerol synthesis, which is also caused, were prevented by adding AICAR, an activator of AMP-activated protein kinase (AMPK), to the incubation medium. These results suggest that hyperglycemia causes insulin resistance in cells other than those in classic insulin target tissues. Whether AMPK activation can reverse or prevent insulin resistance in all of these cells remains to be determined. PMID- 12079835 TI - Insulin stimulation of hepatic triacylglycerol secretion in the insulin-replete state: implications for the etiology of peripheral insulin resistance. AB - Observations on humans, on rats in vivo, and on isolated perfused rat livers indicate that insulin stimulates hepatic very-low-density lipoprotein (VLDL)-TAG secretion when the liver is chronically exposed to the hormone. They suggest that frequent stimulation of insulin secretion throughout the diurnal cycle may result in a chronic stimulation of VLDL secretion and increased delivery of acyl moieties to the periphery, particularly to muscle, the most important site of insulin-sensitive glucose disposal. If acyl groups are provided in excess of the oxidative needs of the tissue, this may lead to induction of insulin resistance, irrespective of whether obesity is established concomitantly. Dietary factors that stimulate hepatic VLDL secretion may have the same effect and contribute to the induction of a vicious spiral leading to the development of the full-blown Metabolic Syndrome and its pathological consequences, including type-2 diabetes, stroke, and cardiovascular disease. PMID- 12079836 TI - Free fatty acid metabolism in human skeletal muscle is regulated by PPARgamma and RXR agonists. AB - Free fatty acid (FFA) oxidation in human skeletal muscle cells can be stimulated, both independently and in a synergistic manner, by agonists for PPARgamma and RXR. Increased FFA disposal in muscle through augmented oxidation could reduce intramyocellular lipid accumulation. The abilities of such agents to improve glucose tolerance and insulin action may thus involve effects on both glucose and FFA metabolism. PMID- 12079838 TI - Adiposity and the development of diabetes in mouse genetic models. AB - While it is certain from the analysis of transgenic models of lipodystrophy that a critical mass of adipose tissue is necessary to prevent the development of diabetes, the reasons why diabetes develops in one obese individual and not in another need to be further investigated. This is also one of the major questions associated with the current obesity epidemic and the development of the metabolic syndrome. The severe lipodystrophic models do not really address this big problem. In this presentation, we point out that models exist among inbred strains of mice that can contribute towards finding answers to the diabetes found in the metabolic syndrome. The differences responsible for the phenotypic variations are undoubtedly multifactorial and involve many genes, but there are powerful genetic resources to investigate these problems. PMID- 12079837 TI - An increase in peroxisomal fatty acid oxidation is not sufficient to prevent tissue lipid accumulation in hHTg rats. AB - We observed earlier that increased skeletal muscle lipid content in the hereditary hypertriglyceridemic (hHTg) rat is accompanied by a decline in plasma leptin. Leptin has recently been shown to enhance peripheral insulin sensitivity by decreasing the tissue triglyceride accumulation, possibly through regulation of fatty acid oxidation and lipogenesis. Thus, to test the hypothesis that insulin resistance and increased skeletal muscle lipid accumulation in hHTg rats are due to a defect in lipid catabolism, we measured mitochondrial and peroxisomal fatty acid oxidation and malonyl-CoA and acetyl-CoA carboxylase-2 content in skeletal muscles of these animals. In addition, we investigated possible molecular mechanisms responsible for the lower leptin levels in hHTg rats by measuring leptin and leptin-receptor (Ob-Ra) mRNA levels. We found the following: (1) in spite of a higher skeletal muscle malonyl-CoA content and an increased sensitivity of carnitine palmitoyltransferase-1 to malonyl-CoA, carnitine palmitoyltransferase-1 activity in muscle of hHTg rats was normal; (2) increased peroxisomal fatty acid oxidation did not seem to be sufficient to prevent the tissue lipid accumulation in these animals; (3) both lower leptin production by white adipose tissue and increased leptin uptake seem to be responsible for lower circulating leptin levels and therefore lower fatty acid catabolism. PMID- 12079839 TI - Modulation of lipid metabolism by energy status of adipocytes: implications for insulin sensitivity. AB - It is becoming evident that insulin resistance of white adipose tissue is a major factor underlying the cardiovascular risk of obesity. Impaired fat storage rather than altered glucose metabolism in adipocytes probably contributes to development of insulin resistance in muscle and other tissues, in particular via increased delivery of nonesterified fatty acids into circulation. Lipid metabolism of adipose tissue is affected by the energy status of fat cells. In vitro experiments indicated the dependence of both lipogenesis and lipolysis on ATP levels in adipocytes. Thus, respiratory uncoupling in adipocytes that results in stimulation of energy dissipation and depression of ATP synthesis may contribute to the control of lipid metabolism, adiposity, and insulin sensitivity. This notion is supported by the expression of UCPs in adipocytes, for example, UCP2, UCP5, as well as some protonophoric anion transporters, and by induction of UCP1 and UCP3 in white fat by pharmacological treatments that reduce adiposity. A negative correlation between expression of UCPs in adipocytes and accumulation of white fat was also found. Expression of UCP1 from the adipose-specific promoter in the aP2-Ucp1 transgenic mice mitigated obesity induced by genetic or dietary factors. The obesity resistance, accompanied by respiratory uncoupling in adipocytes and increased energy expenditure, resulted from ectopic expression of UCP1 in white, but not brown fat. Probably due to depression of the ATP/ADP ratio, both fatty acid synthesis and lipolytic action of norepinephrine in adipocytes of transgenic mice were relatively low. Expression of regulatory G proteins, which are essential for both catecholamine and insulin signaling in adipocytes, was also altered by ectopic UCP1. These results support the role of protonophoric proteins in adipocytes in the control of adiposity and insulin sensitivity. Antidiabetic effects of thiazolidinediones, fibrates, beta(3) adrenoreceptor agonists, dietary n-3 PUFAs, and leptin may be explained at least partially by their effects on the energy and hence also the lipid metabolism of fat cells. PMID- 12079840 TI - Transgenic mice overexpressing the rate-limiting enzyme for hexosamine synthesis in skeletal muscle or adipose tissue exhibit total body insulin resistance. AB - High concentrations of glucose induce insulin resistance and impair insulin secretion in a manner that mirrors type 2 diabetes, a phenomenon known as glucose toxicity. High concentrations of hexosamines mimic these effects, leading to the hypothesis that cells use hexosamine flux as a glucose- and satiety-sensing pathway. Overexpression of the rate-limiting enzyme for hexosamine synthesis (glutamine:fructose-6-phosphate amidotransferase, GFA) in muscle and fat results in insulin resistance and hyperleptinemia. GFA overexpression targeted to liver results in hyperlipidemia and to the beta cell in increased insulin secretion. Thus, excess hexosamine flux leads to a coordinated response whereby fuel is shunted toward long-term storage, mirroring the "thrifty phenotype". The results suggest a mechanism by which chronic overnutrition leads to the phenotype of type 2 diabetes. PMID- 12079841 TI - Enhanced diabetogenic effect of streptozotocin in mice overexpressing UCP-3 in skeletal muscle. AB - Diabetic patients exhibit varying degrees of increased muscle UCP-3 expression in skeletal muscle and, in rodents, the pancreatoxin streptozotocin (STZ) upregulates UCP-3 mRNA in skeletal and cardiac muscles. We have investigated the development of STZ-induced diabetes in transgenic mice overexpressing UCP-3 in skeletal muscle in order to provide further insight on the functional role of muscle UCP-3. UCP-3 transgenic mice treated with STZ (UCP3-STZ) showed a significant increase in blood glucose concentration 3 days after the last dose of STZ with a progressive induction of diabetes, attaining blood glucose concentrations of 24.7 +/- 1.5 mmol/L on day 17. Wild-type mice treated with STZ (WT-STZ) only started to show an increase in blood glucose concentration 6 days after the last dose of STZ and peaked on day 17 at a lower concentration than in the UCP-STZ mice. The pancreatic insulin content of UCP-3 control mice (UCP3-CON) was decreased relative to wild-type control mice (WT-CON), and STZ reduced the total pancreatic insulin content by 72% in WT-STZ mice and by 88% in UCP3-STZ mice. In an insulin tolerance test, blood glucose concentrations declined more in the UCP-3 transgenic mice than in the wild-type mice. Mice overexpressing UCP-3 in skeletal muscle have a lower pancreatic insulin content, but tend to be more insulin-sensitive. These twin actions result in an increased susceptibility to STZ-induced diabetes in UCP-3 transgenic mice. PMID- 12079842 TI - From receptor to effector: insulin signal transduction in skeletal muscle from type II diabetic patients. AB - Insulin resistance is a characteristic feature of type II diabetes mellitus and obesity. Although defects in glucose homeostasis have been recognized for decades, the molecular mechanisms accounting for impaired whole body glucose uptake are still not fully understood. Skeletal muscle constitutes the largest insulin-sensitive organ in humans; thus, insulin resistance in this tissue will have a major impact on whole body glucose homeostasis. Intense efforts are under way to define the molecular mechanisms that regulate glucose metabolism and gene expression in insulin-sensitive tissues. Knowledge of the human genome sequence, used in concert with gene and/or protein array technology, will provide a powerful means to facilitate efforts in revealing molecular targets that regulate glucose homeostasis in type II diabetes mellitus. This will offer quicker ways forward to identifying gene expression profiles in insulin-sensitive and insulin resistant human tissue. This review will present our current understanding of potential defects in insulin signal transduction pathways, with an emphasis on mechanisms regulating glucose transport in skeletal muscle from people with type II diabetes mellitus. Elucidation of the pathways involved in the regulation of glucose homeostasis will offer insight into the causation of insulin resistance and type II diabetes mellitus. Furthermore, this will identify biochemical entry points for drug intervention to improve glucose homeostasis. PMID- 12079843 TI - Skeletal muscle triglycerides: an aspect of regional adiposity and insulin resistance. AB - The composition and biochemistry of skeletal muscle are altered in obesity and type 2 diabetes mellitus (DM) as compared to nonobese individuals. In health, skeletal muscle has a clear capacity to utilize both carbohydrate and lipid fuels and to transition between these in response to hormonal, chiefly insulin, and substrate signals. This metabolic flexibility is key for the major role that skeletal muscle can have in overall fuel balance. In obesity and type 2 DM, there is a loss of this plasticity and, instead, there is metabolic inflexibility. Rates of lipid oxidation do not suppress effectively in response to insulin, but neither do rates of lipid oxidation effectively increase during the transition to fasting conditions. An important morphological characteristic of skeletal muscle in obesity and type 2 DM is an increased content of triglyceride. The accretion of fat within muscle tissues appears to strongly correlate with insulin resistance and may not be simply a passive process, paralleling fat storage in other tissues. Instead, and of particular metabolic interest, a concept is emerging that biochemical characteristics of skeletal muscle in obese individuals dispose to fat accumulation in muscle. An effort to modify skeletal muscle in individuals with obesity and type 2 DM so that the capacity for fat oxidation and metabolic flexibility is improved should be among the goals of treatment for these disorders. PMID- 12079845 TI - Evaluation of free fatty acid metabolism in vivo. AB - In order to enable detailed studies of free fatty acid (FFA) metabolism, we recently introduced a method for the evaluation of tissue-specific FFA metabolism in vivo. The method is based on the simultaneous use of 14C-palmitate (14C-P) and the non-beta-oxidizable FFA analogue, [9,10-3H]-(R)-2-bromopalmitate (3H-R-BrP). Indices of total FFA utilization and incorporation into storage products are obtained from tissue concentrations of 3H and 14C, respectively, following intravenous administration of 3H-R-BrP and 14C-P and their disappearance from plasma into tissues. This review covers the basis for, and developments in, the methodology, as well as some of the applications to date. In the rat, the method has been used to characterize tissue-specific alterations in FFA metabolism in various situations, including skeletal muscle contraction, fasting, hyperinsulinemia, and various pharmacological manipulations. The results of all these studies clearly demonstrate tissue-level control of FFA utilization and metabolic fate, refuting the traditional view that FFA utilization is simply supply-driven. Recent developments enable the simultaneous evaluation of both tissue-specific FFA and glucose metabolism by integrating the use of 2 deoxyglucose and stable isotope-labeled glucose tracers. In conclusion, the 3H-R BrP methodology, especially in combination with other tracers, represents a powerful tool for elucidation of tissue-specific fatty acid metabolism in vivo. PMID- 12079844 TI - Protein kinase C and lipid-induced insulin resistance in skeletal muscle. AB - Insulin resistance of skeletal muscle in humans, animals, and cells is often strongly correlated with increased lipid availability. The elevation of certain intracellular lipid species can lead to the activation of signal transduction pathways that inhibit normal insulin action. Thus, increased diacylglycerol levels in muscle are associated with the activation of one or more isoforms of the protein kinase C family, which is known to attenuate insulin signaling, especially at the level of IRS-1. In addition, de novo synthesis of ceramide can inhibit more distal sites by the activation of protein phosphatase 2A and hence promote the dephosphorylation and inactivation of protein kinase B. Such mechanisms may account at least in part for the reduced insulin sensitivity occurring in obesity and type 2 diabetes where lipid oversupply is a major factor. PMID- 12079846 TI - Insulin action in skeletal muscle: isozyme-specific effects of protein kinase C. AB - Protein kinase C (PKC) is a family of multifunctional isozymes that plays an important role in the regulation of intracellular insulin signal transduction in various insulin-sensitive tissues. This article highlights current understanding on the mechanism of PKC-induced insulin resistance in skeletal muscle, a major target site for insulin-mediated glucose disposal. Initial, apparently contradictory findings on the role of PKC on insulin action can be explained on the basis that certain PKC isoforms (e.g., -zeta and -lambda) have been identified as downstream targets of PI3-kinase activation, while DAG-sensitive PKCs (e.g., -theta; and -epsilon) have negative regulatory effects on insulin signaling. Hence, pharmacological therapies targeting specific PKC isoforms could enhance insulin action and improve glycemic control in patients with impaired glucose tolerance and overt diabetes. PMID- 12079847 TI - Desaturation and elongation of Fatty acids and insulin action. AB - Insulin resistance is characterized by specific changes of the composition of fatty acids in the serum lipids and in the skeletal muscle membranes. Impaired insulin sensitivity is associated with high proportions of palmitic (16:0) acid and low levels of linoleic (18:2 n-6) acid in serum. In addition, there are apparent changes of the fatty acid desaturase activities, suggesting an increased activity of the Delta9 and Delta6 desaturases and a decreased activity of the Delta5 desaturase. The activity of the fatty acid desaturases is regulated by long-chain polyunsaturated fatty acids and insulin and is probably also dependent on the degree of physical activity. A high ratio between arachidonic (20:4 n-6) and dihomo-gamma linolenic (20:3 n-6) acid, as a measure of Delta5 desaturase activity, in the skeletal muscle phospholipids has been related to good insulin sensitivity. Available knowledge seems to indicate that the degree of saturation of the body lipids, and especially the proportion of palmitic acid in the lipid membranes, may be critical for insulin sensitivity. The strong relationships between the Delta5 desaturase activity, a high content of long-chain polyunsaturated fatty acids in the skeletal muscle, and insulin sensitivity may be due to parallel effects of diet and/or physical activity on the fatty acid composition and on insulin sensitivity. PMID- 12079848 TI - Muscle long-chain acyl CoA esters and insulin resistance. AB - A common observation in animal models and in humans is that accumulation of muscle triglyceride is associated with the development of insulin resistance. In animals, this is true of genetic models of obesity and nutritional models of insulin resistance generated by high-fat feeding, infusion of lipid, or infusion of glucose. Although there is a strong link between the accumulation of triglycerides (TG) in muscle and insulin resistance, it is unlikely that TG are directly involved in the generation of muscle insulin resistance. There are now other plausible mechanistic links between muscle lipid metabolites and insulin resistance, in addition to the classic substrate competition proposed by Randle's glucose-fatty acid cycle. The first step in fatty acid metabolism (oxidation or storage) is activation to the long-chain fatty acyl CoA (LCACoA). This review covers the evidence suggesting that cytosolic accumulation of this active form of lipid in muscle can lead to impaired insulin signaling, impaired enzyme activity, and insulin resistance, either directly or by conversion to other lipid intermediates that alter the activity of key kinases and phosphatases. Actions of fatty acids to bind specific nuclear transcription factors provide another mechanism whereby different lipids could influence metabolism. PMID- 12079849 TI - Eicosanoids and the regulation of cardiac glucose transport. AB - Intact actin microfilaments are necessary for insulin-regulated GLUT4 translocation from intracellular pools to the plasma membrane. Products of the lipoxygenase (LO) pathway were shown to be implicated in the regulation of actin cytoskeleton rearrangement. The aim of this study was to examine the role of these LO products for cardiac insulin signaling and glucose uptake, GLUT4 translocation, and actin-based cytoskeleton structure. Exposure of cardiomyocytes to esculetin or NDGA, two structurally different LO inhibitors, induced a complete inhibition of insulin-stimulated glucose uptake, whereas control cells showed a threefold stimulation by insulin. Addition of 12(S)-HETE rendered the NDGA-treated cells insulin-sensitive. Early insulin signaling was not changed in cells exposed to LO inhibitors. Cell surface biotinylation of control cells showed a twofold increase of GLUT4 at the cell surface after insulin stimulation. In contrast, the LO inhibitors induced a complete inhibition of insulin stimulated GLUT4 translocation. Labeling of the F-actin cytoskeleton revealed a prominent disassembly of actin fibers in cells exposed to the LO inhibitors. In conclusion, we show here that products of the LO reaction participate in the organization of the actin network in ventricular cardiomyocytes. Inhibition of LO blocks GLUT4 translocation without affecting insulin signaling events. These data suggest that products of the LO reaction participate in the regulation of glucose transport by contribution to a rearrangement of actin cytoskeletal elements. PMID- 12079850 TI - Regulation of fat metabolism in skeletal muscle. AB - Regulation of carbohydrate and fat utilization by skeletal muscle at rest and during exercise has been the subject of investigation since the early 1960s when Randle et al. proposed the so-called glucose-fatty acid cycle to explain the reciprocal relationship between carbohydrate and fat metabolism. The suggested mechanisms were based on the premise that an increase in fatty acid (FA) availability would result in increased fat metabolism and inhibition of carbohydrate metabolism. Briefly, accumulation of acetyl-CoA would result in inhibition of pyruvate dehydrogenase (PDH), accumulation of citrate would inhibit phosphofructokinase (PFK), and accumulation of glucose-6-phosphate (G6P) would reduce hexokinase (HK) activity. Ultimately, this would inhibit carbohydrate metabolism with increasing availability and oxidation of FA. Although there is some evidence for the existence of the glucose-FA cycle at rest and during low intensity exercise, it cannot explain substrate use at moderate to high exercise intensities. More recently, evidence has accumulated that increases in glycolytic flux may decrease fat metabolism. Potential sites of regulation are the transport of FA into the sarcoplasma, lipolysis of intramuscular triacylglycerol (IMTG) by hormone-sensitive lipase (HSL), and transport of FA across the mitochondrial membrane. There are several potential regulators of fat oxidation: first, malonyl CoA concentration, which is formed from acetyl-CoA, catalyzed by the enzyme acetyl-CoA carboxylase (ACC), which in turn will inhibit carnitine palmitoyl transferase I (CPT I). Another possible mechanism is accumulation of acetyl-CoA that will result in acetylation of the carnitine pool, reducing the free carnitine concentration. This could theoretically reduce FA transport into the mitochondria. There is also some recent evidence that CPT I is inhibited by small reductions in pH that might be observed during exercise at high intensities. It is also possible that FA entry into the sarcolemma is regulated by translocation of FAT/CD36 in a similar manner to glucose transport by GLUT-4. Studies suggest that the regulatory mechanisms may be different at rest and during exercise and may change as the exercise intensity increases. Regulation of skeletal muscle fat metabolism is clearly multifactorial, and different mechanisms may dominate in different conditions. PMID- 12079851 TI - The sphingomyelin-signaling pathway in skeletal muscles and its role in regulation of glucose uptake. AB - Sphingomyelin has been shown to be a source of bioactive compounds. This sphingolipid is located mostly in the outer layer of the plasma membrane and in the membranes of organelles. Sphingomyelin located in the plasma membrane is hydrolyzed into ceramide and phosphorylcholine. Ceramide is the principal second messenger in the sphingomyelin transmembrane signaling pathway. Products of ceramide metabolism, namely, sphingosine, sphingosine-1-phosphate, and ceramide-1 phosphate, also exert broad biological effects. The major effects of ceramide are induction of differentiation, inhibition of proliferation, regulation of inflammatory processes, and induction of apoptosis. There is also convincing evidence that ceramide counteracts insulin-stimulated glucose uptake. Ceramides are also present in skeletal muscles. We investigated ceramide metabolism in different skeletal muscle types of the rat at rest and after prolonged exercise of moderate intensity. Exercise reduced the total content of ceramide fatty acids and changed their composition in each muscle type. These data indicate that the sphingomyelin-signaling pathway functions in skeletal muscles and that its activity is downregulated during prolonged exercise. The content of ceramide in the muscles was inversely related to 2-deoxyglucose uptake by the muscles. This indicates that ceramide may be involved in regulation of glucose uptake by skeletal muscles in vivo. PMID- 12079852 TI - Genetic and molecular analyses of complex metabolic disorders: genetic linkage. AB - Wide efforts have taken place with complex metabolic disorders to emulate the success that linkage analysis has had in explaining the nature of monogenic metabolic diseases such as MODY (maturity-onset diabetes of the young) and FH (familial hypercholesterolemia). New linkage methods are being specifically developed and tested for complex disorders since some of the basic assumptions of traditional linkage analysis used with Mendelian traits are not valid. The nature of complex diseases precludes the use of extended families under the hypothesis that the same disease allele acts in most affected individuals throughout a pedigree. Rather, a multitude of genes and of rare and common alleles creates an apparently chaotic pattern of heterogeneity within and between families. Therefore, very simple family structures, in many studies even isolated sibling pairs, form the basis of efforts to compare the inheritance of disease with that of the chromosomal regions under investigation. Also, assumptions about how individual loci contribute to the overall disease inheritance used for the models applied in linkage computation have to be kept to a minimum. The overall effect of this, together with the potentially weak influence of many loci, is a heavy toll on the statistical power to detect individual contributing genes. This may be the reason why very few scans so far have yielded disease loci that meet genome-wide significance criteria. The confirmation of original loci in secondary studies has proven, as predicted, to be very difficult. Nevertheless, the overall emerging picture is very encouraging: one of the genome scans in type 2 diabetes has been carried through to the positional cloning of the underlying genetic variant, namely, the calpain 10-associated polymorphism in type 2 diabetes. Several other loci have been detected repeatedly throughout studies in various human racial groups, such as the chromosome 1q and 20q diabetes loci, and have become the target of collaborative fine-mapping efforts. Modifications to present methodology are in development with the goal to increase statistical power: examples are the use of intermediate traits with potentially increased genetic homogeneity, the investigation of admixed populations, and the study of linkage disequilibrium over wide genomic regions. PMID- 12079853 TI - Positional cloning of an obesity/diabetes susceptibility gene(s) on chromosome 11 in Pima Indians. AB - Prior results from our genomic scan in Pima Indians indicated an obesity locus in a region on chromosome 11q23-24 that was also linked to diabetes. Bivariate linkage analysis for the combined phenotype "diabesity" gave the strongest evidence for linkage (LOD = 5.2). Our aim is to positionally clone the gene(s) responsible for the linkage. Linkage disequilibrium mapping is being used to narrow the chromosomal region. Single nucleotide polymorphisms (SNPs) are being systematically identified and genotyped at 50-kb intervals across the region of linkage. To date, 455 SNPs have been genotyped in 1229 Pimas. A region containing a cluster of SNPs strongly associated with BMI and a second region, approximately 2 Mb telomeric, containing a cluster of SNPs associated with diabetes have been preliminarily identified. PMID- 12079854 TI - Is a Pro12Ala polymorphism of the PPARgamma2 gene related to obesity and type 2 diabetes mellitus in the Czech population? AB - The peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor subfamily of transcription factors. PPARgamma2 plays a key role in regulation of adipocyte differentiation and energy homeostasis. Recent studies provide evidence that the Pro12Ala polymorphism is linked to obesity and type 2 diabetes mellitus, but the results are controversial and depend on ethnicity. The aim of this study was to determine allele frequencies and to study the influence of the polymorphism on biochemical and anthropometric parameters in a Czech healthy adult population, in type 2 diabetics, and in a group of obese women. RESULTS: The frequency of the Pro12Ala PPARgamma2 gene polymorphism in Czech probands is similar to other central European populations. Frequency of the Pro12Ala substitution tends to be higher in obese women and diabetics compared with controls. The fasting insulin levels in the 12Ala carriers were significantly lower within the group of diabetics even after adjustment for age, BMI, and the length of diabetes duration. In obese women, higher WHR was found in subjects with the 12Ala allele. CONCLUSIONS: This study indicates that the substitution Pro12Ala is not associated with a decreased obesity or diabetes risk in the Czech population. However, the present data show that fasting insulin concentrations are lower in diabetics with the 12Ala allele than in those without it. This finding provides evidence that the polymorphism may influence glucose homeostasis. PMID- 12079855 TI - A custom-built insulin resistance gene chip. AB - OBJECTIVES/AIM: Microarray (gene chip) technology offers a powerful new tool for analyzing the expression of large numbers of genes in many experimental samples. The aim of this study was to design, construct, and use a gene chip to measure the expression levels of key genes in metabolic pathways related to insulin resistance. METHODS: We selected genes that were implicated in the development of insulin resistance, including genes involved in insulin signaling; glucose uptake, oxidation, and storage; fat uptake, oxidation, and storage; cytoskeletal components; and transcription factors. The key regulatory genes in the pathways were identified, along with other recently identified candidate genes such as calpain-10. A total of 242 selected genes (including 32 internal control elements) were sequence-verified, purified, and arrayed on aldehyde-coated slides. RESULTS: Where more than 1 clone containing the gene of interest was available, we chose those containing the genes in the 5' orientation and an insert size of around 1.5 kb. Of the 262 clones purchased, 56 (21%) were found to contain sequences other than those expected. In addition, 2 (1%) did not grow under standard conditions and were assumed to be nonviable. In these cases, alternate clones containing the gene of interest were chosen as described above. The current version of the Insulin Resistance Gene Chip contains 210 genes of interest, plus 48 control elements. A full list of the genes is available at http://www.hbs.deakin.edu.au/mru/research/gene_chip_tech/genechip_three.htm/. CONCLUSIONS: The human Insulin Resistance Gene Chip that we have constructed will be a very useful tool for investigating variation in the expression of genes relevant to insulin resistance under various experimental conditions. Initially, the gene chip will be used in studies such as exercise interventions, fasting, euglycemic-hyperinsulinemic clamps, and administration of antidiabetic agents. PMID- 12079856 TI - Fatty acid regulation of gene expression: a genomic explanation for the benefits of the mediterranean diet. AB - The development of obesity and associated insulin resistance involves a multitude of gene products, including proteins involved in lipid synthesis and oxidation, thermogenesis, and cell differentiation. The genes encoding these proteins are in essence the blueprints that we have inherited from our parents. However, what determines the way in which blueprints are interpreted is largely dictated by a collection of environmental factors. The nutrients we consume are among the most influential of these environmental factors. During the early stages of evolutionary development, nutrients functioned as primitive hormonal signals that allowed the early organisms to turn on pathways of synthesis or storage during periods of nutrient deprivation or excess. As single-cell organisms evolved into complex life forms, nutrients continued to be environmental factors that interacted with hormonal signals to govern the expression of genes encoding proteins involved in energy metabolism, cell differentiation, and cell growth. Nutrients govern the tissue content and activity of different proteins by functioning as regulators of gene transcription, nuclear RNA processing, mRNA degradation, and mRNA translation, as well as functioning as posttranslational modifiers of proteins. One dietary constituent that has a strong influence on cell differentiation, growth, and metabolism is fat. The fatty acid component of dietary lipid not only influences hormonal signaling events by modifying membrane lipid composition, but fatty acids have a very strong direct influence on the molecular events that govern gene expression. In this review, we discuss the influence that (n-9), (n-6), and (n-3) fatty acids exert on gene expression in the liver and skeletal muscle and the impact this has on intra- and interorgan partitioning of metabolic fuels. PMID- 12079857 TI - Perinatal supply and metabolism of long-chain polyunsaturated fatty acids: importance for the early development of the nervous system. AB - The long-chain polyunsaturated fatty acids, arachidonic (AA) and docosahexaenoic acid (DHA), are essential structural lipid components of biomembranes. During pregnancy, long-chain polyunsaturated fatty acids (LC-PUFA) are preferentially transferred from mother to fetus across the placenta. This placental transfer is mediated by specific fatty acid binding and transfer proteins. After birth, preterm and full-term babies are capable of converting linoleic and alpha linolenic acids into AA and DHA, respectively, as demonstrated by studies using stable isotopes, but the activity of this endogenous LC-PUFA synthesis is very low. Breast milk provides preformed LC-PUFA, and breast-fed infants have higher LC-PUFA levels in plasma and tissue phospholipids than infants fed conventional formulas. Supplementation of formulas with different sources of LC-PUFA can normalize LC-PUFA status in the recipient infants relative to reference groups fed human milk. Some, but not all, randomized, double-masked placebo-controlled clinical trials in preterm and healthy full-term infants demonstrated benefits of formula supplementation with DHA and AA for development of visual acuity up to 1 year of age and of complex neural and cognitive functions. From the available data, we conclude that LC-PUFA are conditionally essential substrates during early life that are related to the quality of growth and development. Therefore, a dietary supply during pregnancy, lactation, and early childhood that avoids the occurrence of LC-PUFA depletion is desirable, as was recently recommended by an expert consensus workshop of the Child Health Foundation. PMID- 12079858 TI - The responses of serum and adipose Fatty acids to a one-year weight reduction regimen in female obese monozygotic twins. AB - We have reported strong intrapair resemblances (IPRs) in serum phosphatidylcholine (PC) fatty acid composition within adult monozygotic twins living apart. This study assessed the contribution of genetic factors to changes in serum and adipose tissue fatty acids resulting from weight loss and followed by a subsequent year of weight maintenance. Eleven pairs of female obese monozygotic twins (age: 38.9 +/- 1.8; BMI: 32.5 +/- 0.9) were recruited for the study. Fasting serum and adipose tissue were obtained after 1 week of inpatient stabilization, after 1 month of inpatient very-low-calorie diet (VLCD), and again after 1 year of outpatient weight maintenance. Fatty acids in serum lipid fractions and adipose tissue were quantitated by gas chromatography. Using multiple regression adjusted for age and initial value, IPRs were determined for the changes induced by VLCD and by the year of weight maintenance. There were few IPRs in nonessential fatty acids. By contrast, there were numerous IPRs for essential fatty acids (EFA), especially in the n-3 family across the VLCD. Following the maintenance year, however, frequent IPRs for nonessential fatty acids were seen, particularly in serum PC, and strong IPRs were seen for 18:3 n-3 and 20:5 n-3 across multiple fractions. These results infer the existence of strong genetic factors determining both the nonessential and EFA compositions of tissue lipids in humans independent of diet. Of particular note were the consistent IPRs for n-3 fatty acids despite dietary stress, indicating that the conservation and distribution of this EFA family are subject to considerable genetic variance in humans. PMID- 12079859 TI - Dietary fat as a risk factor for type 2 diabetes. AB - Too much information is missing for a definitive recommendation to be made concerning the role of diet in the development of diabetes. Although studies in certain population subgroups show a positive correlation between amount of dietary fat intake and occurrence of type 2 diabetes, these cannot be considered definitive, in part because they are confounded by many other variables that might relate to the development of diabetes. Furthermore, dietary fat intake has not been correlated with certainty to diabetes risk factors. With regard to diabetes risk, type of dietary fat consumed may be more important than total dietary fat intake. Several studies suggest that diets higher in saturated fat may pose a higher diabetes risk than those higher in unsaturated fat. Although there is a general consensus that reduced consumption of saturated fat and cholesterol reduces CVD risk, there are no definite nutrition recommendations to prevent diabetes. The most important recommendation may be for health-care providers to carefully assess diabetes risk in their patients and institute dietary changes on an individual basis as needed to achieve and maintain body weight. PMID- 12079860 TI - Type of dietary fat and insulin resistance. AB - Animal studies have already shown the possibility to modulate insulin action by changing not only the amount of total fat, but also the type of fat. In these studies, saturated fat significantly increased insulin resistance, long- and short-chain omega(3) fatty acids significantly improved it, whereas the effects of monounsaturated and omega(6) polyunsaturated fatty acids ranged somewhere in between the two. A recent multicenter study (the Kanwu study) on humans has shown that shifting from a diet rich in saturated fatty acids to one rich in monounsaturated fat improved insulin sensitivity in healthy people, while a moderate omega(3) supplementation did not affect it; this second finding confirms previous results in type 2 diabetic patients with hypertriglyceridemia. There are also other aspects of the metabolic syndrome that can be influenced by the different type of dietary fat, particularly blood pressure and lipid metabolism. With respect to blood pressure, the majority of studies show that omega(3) fatty acids are able to reduce blood pressure in hypertensive patients, but not in normotensive individuals; this result has been confirmed also by the Kanwu study, where no changes in blood pressure were seen after omega(3) supplementation in healthy people. On the other hand, in this study, the change from saturated to monounsaturated fatty acids was able to significantly reduce diastolic blood pressure. As to the lipid abnormalities more frequently present in the metabolic syndrome (i.e., hypertriglyceridemia and low HDL cholesterol), the main effects are related to omega(3) fatty acids, which surely reduce triglyceride levels, but at the same time increase LDL cholesterol. In conclusion, there is so far sound evidence in humans that the quality of dietary fat is able to influence insulin resistance and some of the related metabolic abnormalities. PMID- 12079861 TI - Treatment of hypertriglyceridemia with fenofibrate, fatty acid composition of plasma and LDL, and their relations to parameters of lipoperoxidation of LDL. AB - The purpose of this study was to determine oxidation and oxidability of VLDL and LDL in connection with changes in their composition and content of FA in LDL after treatment with fenofibrate in patients with HTG. PMID- 12079862 TI - Effect of iron depletion on cardiovascular risk factors: studies in carbohydrate intolerant patients. AB - Controversy surrounds the role of iron (Fe) in atherosclerosis (ASCVD), mainly due to the inaccuracy of assessing body Fe stores with serum ferritin and transferrin saturation. Quantitative phlebotomy was used to test whether or not (a) Fe stores are increased in individuals at high risk for ASCVD and (b) Fe depletion to near-deficiency (NID) levels is associated with reduction of risk factors for ASCVD. Thirty-one carbohydrate-intolerant subjects completed the study. Fe stores were within normal limits (1.5 +/- 0.1 g). At NID, a significant increase of HDL-cholesterol (p < 0.001) and reductions of blood pressure (p < 0.001), total and LDL-cholesterol (p < 0.001), triglyceride (p < 0.001), fibrinogen (p < 0.001) and glucose and insulin responses to oral glucose loading (p < 0.001) were noted, while homocysteine plasma concentration remained unchanged. These effects were largely reversed by a 6-month period of Fe repletion with reinstitution of Fe sufficiency. Thus, although individuals at high risk for ASCVD are not Fe-overloaded, they seem to benefit, metabolically and hemodynamically, from lowering of body Fe to levels commonly seen in premenopausal females. PMID- 12079863 TI - Erythrocyte membrane ion transport in offspring of hypertensive parents: effect of acute hyperinsulinemia and relation to insulin action. AB - Some patients with essential hypertension exhibit insulin resistance (IR) and several red blood cell (RBC) ion transport abnormalities. The aims of the study were to assess RBC ion transport acitivities under basal conditions, to test in vivo the effect of acute hyperinsulinemia, and to evaluate the relationship to IR in the offspring of hypertensive parents (n = 12; OHP) and healthy controls (n = 14; C). Activities of the Na+-K+ pump, Na+-K+ cotransport, Na+-Li+ countertransport (SLC), and Na+, Rb+, and Li+ leaks (passive membrane permeability) were measured before and after a hyperinsulinemic (75 microU/mL) euglycemic clamp (HIC) and compared to those found under isoinsulinemic isovolumic conditions in OHP and C. An insulin action was calculated as glucose disposal and insulin sensitivity index (M/I) after HIC. OHP were characterized by lower M/I (0.12+/-0.07 vs. 0.20+/-0.09 mg/kg/min/microU/mL; p < 0.05) and elevated SLC and Li+ and Rb+ leaks (p < 0.05) compared with C. Although acute hyperinsulinemia did not modify significantly any ion transport parameter studied, negative correlation was observed between insulin action and membrane cation leaks. Glucose disposal correlated with an Li+ leak in C (r = -0.736; p < 0.01) and all subjects (r = -0.424; p < 0.05) after HIC and in OHP with an Na+ leak (r = -0.727; p < 0.05) before HIC. In conclusion, OHP displayed higher insulin resistance, enhanced activity of SLC, and augmented Li+ and Rb+ leaks. Acute hyperinsulinemia did not modify any ion transport parameter studied, although negative correlation was observed between insulin action and membrane leaks. PMID- 12079864 TI - Increased fat intake, impaired fat oxidation, and failure of fat cell proliferation result in ectopic fat storage, insulin resistance, and type 2 diabetes mellitus. AB - It is widely accepted that increasing adiposity is associated with insulin resistance and increased risk of type 2 diabetes. The predominant paradigm used to explain this link is the portal/visceral hypothesis. This hypothesis proposes that increased adiposity, particularly in the visceral depots, leads to increased free fatty acid flux and inhibition of insulin action via Randle's effect in insulin-sensitive tissues. Recent data do not entirely support this hypothesis. As such, two new paradigms have emerged that may explain the established links between adiposity and disease. (A) Three lines of evidence support the ectopic fat storage syndrome. First, failure to develop adequate adipose tissue mass in either mice or humans, also known as lipodystrophy, results in severe insulin resistance and diabetes. This is thought to be the result of ectopic storage of lipid into liver, skeletal muscle, and the pancreatic insulin-secreting beta cell. Second, most obese patients also shunt lipid into the skeletal muscle, the liver, and probably the beta cell. The importance of this finding is exemplified by several studies demonstrating that the degree of lipid infiltration into skeletal muscle and liver correlates highly with insulin resistance. Third, increased fat cell size is highly associated with insulin resistance and the development of diabetes. Increased fat cell size may represent the failure of the adipose tissue mass to expand and thus to accommodate an increased energy influx. Taken together, these three observations support the acquired lipodystrophy hypothesis as a link between adiposity and insulin resistance. (B) The endocrine paradigm developed in parallel with the ectopic fat storage syndrome hypothesis. Adipose tissue secretes a variety of endocrine hormones, such as leptin, interleukin-6, angiotensin II, adiponectin (also called ACRP30 and adipoQ), and resistin. From this viewpoint, adipose tissue plays a critical role as an endocrine gland, secreting numerous factors with potent effects on the metabolism of distant tissues. These two new paradigms provide a framework to advance our understanding of the pathophysiology of the insulin-resistance syndrome. PMID- 12079865 TI - Leptin signaling, adiposity, and energy balance. AB - A chronic minor imbalance between energy intake and energy expenditure may lead to obesity. Both lean and obese subjects eventually reach energy balance and their body weight regulation implies that the adipose tissue mass is "sensed", leading to appropriate responses of energy intake and energy expenditure. The cloning of the ob gene and the identification of its encoded protein, leptin, have provided a system signaling the amount of adipose energy stores to the brain. Leptin, a hormone secreted by fat cells, acts in rodents via hypothalamic receptors to inhibit feeding and increase thermogenesis. A feedback regulatory loop with three distinct steps has been identified: (1) a sensor (leptin production by adipose cells) monitors the size of the adipose tissue mass; (2) hypothalamic centers receive and integrate the intensity of the leptin signal through leptin receptors (LRb); (3) effector systems, including the sympathetic nervous system, control the two main determinants of energy balance-energy intake and energy expenditure. While this feedback regulatory loop is well established in rodents, there are many unsolved questions about its applicability to body weight regulation in humans. The rate of leptin production is related to adiposity, but a large portion of the interindividual variability in plasma leptin concentration is independent of body fatness. Gender is an important factor determining plasma leptin, with women having markedly higher leptin concentrations than men for any given degree of fat mass. The ob mRNA expression is also upregulated by glucocorticoids, whereas stimulation of the sympathetic nervous system results in its inhibition. Furthermore, leptin is not a satiety factor in humans because changes in food intake do not induce short-term increases in plasma leptin levels. After its binding to LRb in the hypothalamus, leptin stimulates a specific signaling cascade that results in the inhibition of several orexigenic neuropeptides, while stimulating several anorexigenic peptides. The orexigenic neuropeptides that are downregulated by leptin are NPY (neuropeptide Y), MCH (melanin-concentrating hormone), orexins, and AGRP (agouti related peptide). The anorexigenic neuropeptides that are upregulated by leptin are alpha-MSH (alpha-melanocyte-stimulating hormone), which acts on MC4R (melanocortin-4 receptor); CART (cocaine and amphetamine-regulated transcript); and CRH (corticotropin-releasing-hormone). Obese humans have high plasma leptin concentrations related to the size of adipose tissue, but this elevated leptin signal does not induce the expected responses (i.e., a reduction in food intake and an increase in energy expenditure). This suggests that obese humans are resistant to the effects of endogenous leptin. This resistance is also shown by the lack of effect of exogenous leptin administration to induce weight loss in obese patients. The mechanisms that may account for leptin resistance in human obesity include a limitation of the blood-brain-barrier transport system for leptin and an inhibition of the leptin signaling pathways in leptin-responsive hypothalamic neurons. During periods of energy deficit, the fall in leptin plasma levels exceeds the rate at which fat stores are decreased. Reduction of the leptin signal induces several neuroendocrine responses that tend to limit weight loss, such as hunger, food-seeking behavior, and suppression of plasma thyroid hormone levels. Conversely, it is unlikely that leptin has evolved to prevent obesity when plenty of palatable foods are available because the elevated plasma leptin levels resulting from the increased adipose tissue mass do not prevent the development of obesity. In conclusion, in humans, the leptin signaling system appears to be mainly involved in maintenance of adequate energy stores for survival during periods of energy deficit. Its role in the etiology of human obesity is only demonstrated in the very rare situations of absence of the leptin signal (mutations of the leptin gene or of the leptin receptor gene), which produces an internal perception of starvation and results in a chronic stimulation of excessive food intake. PMID- 12079866 TI - Neuroimaging and obesity: mapping the brain responses to hunger and satiation in humans using positron emission tomography. AB - The hypothalamus has a major role in the control of food intake. However, neurotracing studies have shown that the hypothalamus receives input from several other regions of the brain that are likely to modulate its activity. Of particular interest to the understanding of human eating behavior is the possible involvement of the cortex. Using positron emission tomography (PET), we generated functional brain maps of the neuroanatomical correlates of hunger (after a 36-h fast) and satiation (after oral administration of a liquid formula meal) in lean and obese subjects. Results in lean individuals indicate that the neuroanatomical correlates of hunger form a complex network of brain regions including the hypothalamus, thalamus, and several limbic/paralimbic areas such as the insula, hippocampal/parahippocampal formation, and the orbitofrontal cortex. Satiation was associated with preferentially increased neuronal activity in the prefrontal cortex. Our studies also indicate that the brain responses to hunger/satiation in the hypothalamus, limbic/paralimbic areas (commonly associated with the regulation of emotion), and prefrontal cortex (thought to be involved in the inhibition of inappropriate response tendencies) might be different in obese and lean individuals. In conclusion, neuroimaging of the human brain is proving to be an important tool for understanding the complexity of brain involvement in the regulation of eating behavior. PET studies might help to unravel the neuropathophysiology underlying human obesity. PMID- 12079867 TI - Uncoupling proteins, leptin, and obesity: an updated review. AB - The hypothesis that the novel uncoupling protein UCP3 is thermogenic and/or thermoregulatory is discussed. In vitro, ex vivo, and in vivo models are presented. The beta(3)-adrenoceptors are crucial for the appearance of UCP1 expressing cells in the white adipose tissue. These cells might differ from classical brown adipocytes. Besides its well-known effect on brown adipose tissue UCP1, leptin might stimulate indirectly muscle UCP3 expression. PMID- 12079868 TI - New approaches to gene discovery with animal models of obesity and diabetes. AB - DNA-based approaches to the discovery of genes contributing to the development of type 2 diabetes have not been very successful despite substantial investments of time and money. The multiple gene-gene and gene-environment interactions that influence the development of type 2 diabetes mean that DNA approaches are not the ideal tool for defining the etiology of this complex disease. Gene expression based technologies may prove to be a more rewarding strategy to identify diabetes candidate genes. There are a number of RNA-based technologies available to identify genes that are differentially expressed in various tissues in type 2 diabetes. These include differential display polymerase chain reaction (ddPCR), suppression subtractive hybridization (SSH), and cDNA microarrays. The power of new technologies to detect differential gene expression is ideally suited to studies utilizing appropriate animal models of human disease. We have shown that the gene expression approach, in combination with an excellent animal model such as the Israeli sand rat (Psammomys obesus), can provide novel genes and pathways that may be important in the disease process and provide novel therapeutic approaches. This paper will describe a new gene discovery, beacon, a novel gene linked with energy intake. As the functional characterization of novel genes discovered in our laboratory using this approach continues, it is anticipated that we will soon be able to compile a definitive list of genes that are important in the development of obesity and type 2 diabetes. PMID- 12079869 TI - High-fat high-energy feeding impairs fasting glucose and increases fasting insulin levels in the Gottingen minipig: results from a pilot study. AB - High-fat diet and obesity are known to be of major importance for development of type 2 diabetes in humans. High-fat feeding can induce syndromes of glucose intolerance and/or insulin resistance in several species, and the Gottingen minipig might be a useful model for studying the effect of dietary high-fat intake and obesity on glucose homeostasis and the susceptibility to diabetes. The present study was designed as a pilot study to investigate the effects of obesity caused by high-fat high-energy feeding on oral and intravenous glucose tolerance. Male Gottingen minipigs were fed a control diet (CD) or a high-fat high-energy diet (HFD) for 3 months. Body weight (32.6 +/- 2.4 kg vs. 24.9 +/- 0.5 kg, p < 0.001) and total (13.2 +/- 3.2% vs. 6.1 +/- 0.5%, p = 0.002) and truncal (11.0 +/ 3.9% vs. 1.8 +/- 1.1%, p = 0.001) fat percent were increased significantly, whereas relative lean body mass was decreased (84.8 +/- 3.3% vs. 91.9 +/- 0.5%, p = 0.002) in the HFD group compared to CD. Fasting plasma glucose (4.3 +/- 0.4 mM vs. 3.6 +/- 0.3 mM, p = 0.023) and insulin (80 +/- 23 pM vs. 23 +/- 21 pM, p = 0.012) were increased in the HFD group compared to CD, but oral glucose tolerance was not significantly changed. Insulin responses to intravenous glucose were increased (6741 +/- 2538 vs. 3938 +/- 771 pM 3 min, p = 0.050), while glucose clearance was not changed by HFD vs. CD, thus indicating insulin resistance. In conclusion, changes in body weight and composition, resulting in minor abnormalities in glucose tolerance and insulin sensitivity, characterized by slight hyperglycemia and compensatory hyperinsulinemia, can be induced in the male Gottingen minipig by high-fat high-energy feeding for 3 months. This approach seems to be an interesting and promising method for establishment of a nonrodent model of insulin resistance or type 2 diabetes. PMID- 12079871 TI - Opposing effects of fatty acids and acyl-CoA esters on conformation and cofactor recruitment of peroxisome proliferator-activated receptors. AB - The peroxisome proliferator-activated receptors (PPARs) bind and are activated by a variety of fatty acids and derivatives thereof. Agonist binding enhances PPAR mediated transactivation via release of corepressors and recruitment of coactivator complexes. Recently, we and others have reported that acyl-CoA esters act as PPAR antagonists in vitro. Here, we show that the binding of the nonhydrolyzable acyl-CoA analogue, S-hexadecyl-CoA, differentially affected conformation and coactivator recruitment of the individual PPAR subtypes. In protease protection assays, S-hexadecyl CoA increased the sensitivity of PPARalpha and PPARdelta towards chymotrypsin, whereas the action of chymotrypsin on PPARgamma was only marginally affected, suggesting distinct subtype-dependent differences in the effects of S-hexadecyl-CoA on conformation of the PPARs. In keeping with these findings, S-hexadecyl-CoA abrogated ligand-induced recruitment of coactivators to PPARalpha and PPARdelta, whereas coactivator recruitment to PPARgamma was unaffected by S-hexadecyl-CoA. PMID- 12079872 TI - Relationship between insulin resistance and muscle triglyceride content in nonobese and obese experimental models of insulin resistance syndrome. AB - In nonobese hereditary hypertriglyceridemic (HTg) rats and obese HTg Koletsky SHROB/Kol rats, muscle triglyceride accumulation and its relationship to glucose homeostasis and tissue sensitivity to insulin action were examined. Soleus muscle and diaphragm triglyceride contents were markedly increased in HTg rats as compared with controls and were further increased in obese animals. On the other hand, glucose intolerance and impairment of insulin-stimulated in vitro glycogen synthesis were of a similar degree in nonobese as well as obese animals. Results indicate that insulin resistance did not increase proportionally to muscle triglyceride content. PMID- 12079870 TI - Comparison of the extrapancreatic action of BRX-220 and pioglitazone in the high fat diet-induced insulin resistance. AB - A new Biorex molecule, BRX-220, has been shown to be effective in animal models of diabetic neuro- and retinopathy. Recent in vitro studies showed that it might also have an insulin-sensitizing action. Therefore, the effect of BRX-220 on insulin sensitivity was compared with the action of pioglitazone (PGZ) in high fat (HF) diet-induced insulin resistance (IR) of rats. METHODS: Male Wistar rats were fed for 3 weeks a standard chow (PD) or the HF (70-cal%) diet. The HF-fed rats were also given daily BRX-220 (20 mg/kg BW) or PGZ (6 mg/kg BW) by gavage. In vivo insulin action was assessed by the euglycemic hyperinsulinemic clamp. Glucose, insulin, FFA, triglyceride (TG), and glycerol levels in blood were also measured, as well as tissue TG content. RESULTS: Increased levels of fed TG in circulation after HF diet (PD: 2.0+/-0.2 vs. HF: 5.0+/-0.8 mmol/L) were partially corrected by BRX-220 (HF + BRX: 3.8+/-0.3) and normalized by PGZ (HF + PGZ: 2.6+/ 0.3). Both molecules prevented the increase in fed serum FFA levels after HF diet (PD: 0.5+/-0.06; HF: 1.8+/-0.2 mmol/L), with a more pronounced effect of PGZ (HF + BRX: 1.2+/-0.1; HF + PGZ: 0.7+/-0.06). Tissue TG levels increased significantly in response to HF feeding in both liver (HF: 16+/-3.0; PD: 6.4+/-1.1 micromol/g) and skeletal muscle (HF: 7.7+/-1.2; PD: 2.4+/-0.4). This increase was completely normalized by both agents in the liver (HF + BRX: 8.8+/-0.8; HF + PGZ: 8.8+/ 1.0), and only partially in the skeletal muscles. HF diet-induced in vivo IR (PD: 25.4+/-0.5; HF: 15.7+/-0.5 mg/kg/min) was significantly reduced by BRX-220 (HF + BRX: 18.7+/-0.3) and PGZ (HF + PGZ: 22.8+/-0.4) treatment. CONCLUSIONS: (1) Subchronic administration of BRX-220 leads to an improvement of in vivo insulin action. (2) This insulin-sensitizing effect is, however, not as pronounced as that of PGZ. (3) It is accompanied by a decrease of circulating TG and FFA levels in the postprandial state and (4) by lower TG content in liver and skeletal muscle. PMID- 12079873 TI - Insulin resistance in the hereditary hypertriglyceridemic rat is associated with an impairment of delta-6 desaturase expression in liver. AB - Our previous studies have shown that insulin resistance (IR) in the hereditary hypertriglyceridemic (hHTg) rat is accompanied by a specific fatty acid (FA) profile in insulin target tissues, possibly due to a defect in the desaturation pathway. Increased dietary intake of n-3 polyunsaturated fatty acids (PUFAs) was shown to shape FA composition and to improve insulin sensitivity in this animal strain. Thus, the aim of this study is twofold: (1) to evaluate a defect in the FA desaturation by direct measurement of enzyme activity and gene expression for Delta-6 desaturase (Delta-6 D) in liver of hHTg rats and (2) to investigate the effect of dietary n-3 PUFAs on hepatic Delta-6 D in relation to tissue FA composition. Male Wistar or hHTg rats were fed ad libitum for 21 days either the basal or fish oil (FO)-supplemented diets. Triglyceride (Tg) levels in serum and tissue lipid extracts were measured with the aid of a commercially available enzymatic set. Hepatic activity of the Delta-6 D was determined radiometrically in a microsomal fraction using 1-(14)C-linoleic acid as a substrate. The Delta-6 D mRNA levels were measured using the Northern blot technique. Tissue FA composition was determined by gas chromatography in the total phospholipid fraction after TLC separation. Increased levels of Tg in hHTg rat circulation were accompanied by raised accumulation of Tg in skeletal muscles. FO feeding lowered the concentration of Tg in serum and prevented their accumulation in skeletal muscles of hHTg rats. A pronounced decrease in the hepatic Delta-6 D activity in hHTg rats (by about 80%) was not further diminished by FO feeding. On the other hand, the activity of Delta-6 D in liver of control rats was reduced by about 40% after FO supplementation. These changes were paralleled by a decrease in the Delta-6 D index as calculated from the liver phospholipid FA profile. In particular, an increase in the amount of 18:2 n-6 and a decrease in arachidonic acid and PUFA n-6 metabolites were found. The results indicate that a decrease of insulin action in hHTg rats is accompanied by an impairment of the hepatic Delta 6 D activity already at the gene level, which is not further affected by n-3 PUFA supplementation. PMID- 12079874 TI - Heart remodeling in the hereditary hypertriglyceridemic rat: effect of captopril and nitric oxide deficiency. AB - AIM: The hereditary hypertriglyceridemic (hHTg) rat is characterized by insulin resistance, hypertension, and hypertriglyceridemia. Thus, we investigated whether (a) remodeling of the heart left ventricle (LV) is present under the given hypertensive situation and (b) whether this potential alteration could be influenced by an inhibition of the angiotensin converting enzyme (ACE) and/or by a blockade of nitric oxide production. METHODS: Five groups of rats were investigated: control Wistar (C) rats, hHTg rats, hHTg rats given captopril (100 mg/kg/day) (hHTg + CAP) or NG-nitro-l-arginine methyl ester (L-NAME, 40 mg/kg/day) (hHTg + L-NAME), and hHTg rats given the combination of both drugs (hHTg + CAP + L-NAME) for 28 days. Systolic blood pressure (SBP) was measured by tail-cuff plethysmography each week. After cervical dislocation, the relative weights of the left and right ventricles (LV/BW, RV/BW) were obtained, the LV nucleic acid concentrations were analyzed, and the fibrosis amount was quantified with aid of a semiquantitative histological technique. RESULTS: In the hHTg group, the increased SBP (141.7 +/- 4.4 vs. 117.2 +/- 3.1 mmHg in controls) was linked to hypertrophy of the LV (1.63 +/- 0.05 vs. 1.30 +/- 0.03 g/kg in controls) with only a minimum of fibrosis. DNA concentration in the LV was decreased (0.45 +/- 0.03 vs. 0.69 +/- 0.04 mg/g w.w. in controls) in the hHTg group. Captopril normalized SBP and decreased the LV/BW (1.44 +/- 0.04 g/kg). Chronic administration of L-NAME to the hHTg rats additionally enhanced (189.3 +/ 5.9 mmHg) the already raised SBP, stimulated fibrosis development, and increased DNA concentration (0.54 +/- 0.02 mg/g w.w.) in the LV compared to hHTg group, yet without additional weight increase of the LV. The combined treatment of the hHTg rats with CAP and L-NAME resulted in normal SBP and the development of LV hypertrophy, and fibrosis was substantially reduced. CONCLUSIONS: (a) The heart of hHTg rats carries signs of LV hypertrophy with minimal fibrosis. (b) Nevertheless, LV fibrosis was increased in the hHTg + L-NAME group. (c) Captopril normalized SBP and decreased the extent of LV hypertrophy in both the nontreated hHTg and the hHTg + L-NAME groups and (d) substantially reduced the development of LV fibrosis in the hHTg + L-NAME group. LVH in hHTg rats may be induced by sympathoadrenal system activation, circulating volume enlargement, and impairment of nitric oxide (NO) production rather than by activation of the renin angiotensin-aldosterone system. PMID- 12079875 TI - Energy transfer in acute diabetic rat hearts: adaptation to increased energy demands due to augmented calcium transients. AB - OBJECTIVES: Hearts of rats with diabetes mellitus (DM) are characterized by energy demands exceeding their energy production, but they might also exhibit decreased vulnerability to ischemia and calcium overload. This indicates adaptation in cardiac energetics (CE), where energy transport is not rate limiting. Aim-This study was designed to elucidate the functional significance of the DM-induced adaptation in CE by investigating the formation of mitochondrial contact sites (MiCS), facilitating the Ca-dependent/high-capacity energy transfer from mitochondria, in conjunction with testing the ischemic tolerance (IT) of hearts. METHODS: After 1 week of streptozotocin-induced DM (45 mg/kg iv), the hearts of male diabetic and age-matched control rats (C) were isolated and Langendorff-perfused with either 1.6 or 2.2 mmol/L of CaCl(2). MiCS formation was assessed by cytochemical detection of mCPK octamers and was quantified stereologically as MiCS to mitochondrial surface ratio (S(S)). IT was evaluated in anesthetized open-chest animals subjected to 30-min occlusion of the LAD coronary artery followed by 4-h reperfusion, by monitoring ischemic arrhythmias and by measuring the size of infarction (tetrazolium double staining). RESULTS: In C hearts, increasing Ca2+ induced both positive inotropic response (dP/dt increase from 2270 +/- 220 to 2955 +/- 229, p < 0.01) and elevated MiCS formation (S(S) increase from 0.070 +/- 0.011 to 0.123 +/- 0.012, p < 0.01). In DM hearts, basic MiCS formation was already comparable with that induced by elevated Ca2+ in C hearts and could not be further stimulated by Ca2+. In C, ventricular tachycardia represented 55.4% of the total arrhythmias and occurred in 90% of the animals. In DM rats, the arrhythmia profile was similar to that in C, and the incidence of tachyarrhythmias and their severity were not enhanced (arrhythmia score: 3.18 +/- 0.4 vs. 3.30 +/- 0.3 in C). The infarct size normalized to the size of area at risk was smaller in the DM than in C hearts (52.3 +/- 5.8% vs. 69.2 +/- 2.2%, respectively; p < 0.05). CONCLUSIONS: Ca-signaling represents the link between energy delivery from mitochondria (via MiCS) and energy requirements of the heart. In DM hearts, energy transport via MiCS is elevated to the maximum value. This contributes to increased resistance of DM hearts to irreversible cell damage. PMID- 12079876 TI - Impaired endothelial function of thoracic aorta in hereditary hypertriglyceridemic rats. AB - OBJECTIVE: Hereditary hypertriglyceridemia (hHTG) in rats was found to be associated with metabolic abnormalities and elevation of blood pressure. There is controversy regarding the relation between hHTG and vascular function. The aim of this study was to determine the reactivity and accompanying structural changes in thoracic aorta from hereditary hypertriglyceridemic rats and hHTG rats that were given, for a long time, N(G)-nitro-l-arginine methyl ester (L-NAME) with and without simultaneous captopril treatment. METHODS: Isolated rings of thoracic aorta were mounted in organ chambers for isometric tension recording or for measurement of endothelium-dependent relaxation. Morphological changes of thoracic aorta (wall thickness, diameter) were measured using light microscopy. RESULTS: Endothelium-dependent relaxation (EDR) to acetylcholine (ACh, 10(-5) M) was significantly attenuated in the hHTG group compared to control Wistar rats (59.3 +/- 8.5% vs. 95.8 +/- 6.5%, p < 0.001), but normalized after pretreatment with captopril. EDR to ACh was further inhibited in hHTG rats treated with L-NAME (36.0 +/- 2.3%, p < 0.001). Maximum residual relaxation was only partly restored with captopril treatment (72.4 +/- 5.8%, p < 0.001). Hypertriglyceridemia did not significantly alter the sensitivity of the thoracic aorta to exogenous noradrenaline. The diameter/wall thickness (D/W) ratio in aortas of control Wistar rats averaged 16.25 +/- 0.57. This ratio was significantly lower in hHTG rats (12.52 +/- 0.38, p < 0.01) and was not altered after treatment with captopril. In the hHTG rats treated with L-NAME, the D/W ratio was further significantly decreased (8.25 +/- 0.30, p < 0.001). Simultaneous captopril treatment attenuated the decrement of this ratio (9.80 +/- 0.75, p < 0.05). CONCLUSIONS: Results showed that hHTG is accompanied by functional and morphological alterations in the rat thoracic aorta. These changes in hHTG and in hHTG rats treated with L-NAME could be, at least in part, protected by captopril treatment. PMID- 12079877 TI - A novel Mammalian homologue of a bacterial citrate-metabolizing enzyme. AB - Mammals metabolize citrate to acetyl-CoA and oxaloacetate via the enzyme, ATP:citrate lyase. Bacteria lack this enzyme, but have the ability to cleave citrate in the form of citryl-CoA in an analogous manner using a structurally distinct enzyme. We have identified a novel mammalian gene that shows significant amino acid sequence homology to the bacterial CitE gene product that is responsible for cleavage of citryl-CoA. We propose that this gene encodes an enzyme that catalyzes cleavage of substrates related to CoA esters of citrate or an analogous intermediary metabolite. The product of this novel gene may represent a component of an unknown metabolic pathway in mammals. PMID- 12079878 TI - Effect of BRX-220 against peripheral neuropathy and insulin resistance in diabetic rat models. AB - Bimoclomol (BML), a symptomatic antidiabetic agent, has been developed by Biorex R & D Co. to treat diabetic neuropathy and retinopathy. BRX-220, an orally active member of the BRX family, has been developed to treat diabetic complications and insulin resistance (IR) as a follow-up compound. The effect of BRX-220 on peripheral neuropathy was examined in rats with diabetes (type 1) induced by administration of a beta-cell toxin, streptozotocin (STZ, 45 mg/kg iv). Nerve functions were evaluated by electrophysiological measurements of muscle motor and sensory nerve conduction velocities (MNCV and SNCV, respectively). MNCV and SNCV decreased in diabetic rats by 25% (p < 0.001). A 1-month preventive treatment with BRX-220 (2.5, 5, 10, and 20 mg/kg po) dose-dependently improved diabetes related deficits in MNCV (51.3%, 71.3%, 86.1%, and 91.3%) and SNCV (48.9%, 68.5%, 86.1%, and 93.2%). Insulin sensitivity was measured using the insulin tolerance test (ITT), both in STZ diabetic and in Zucker diabetic fatty (ZDF) rats (model of type 2 diabetes). Severe IR was detected in STZ diabetic and ZDF rats. This resistance was significantly (p < 0.05) reduced by BRX-220 treatment. PMID- 12079879 TI - Terguride treatment attenuated prolactin release and enhanced insulin receptor affinity and GLUT 4 content in obese spontaneously hypertensive female, but not male rats. AB - Glucose tolerance, serum insulin, insulin receptors in epididymal fat tissue, and GLUT 4 content in muscle, as well as serum prolactin, were studied in obese and lean spontaneously hypertensive rats (SHRs) of both sexes. Obese animals displayed insulin resistance and decreased capacity of high-affinity binding sites of insulin receptors in fat tissue plasma membranes. GLUT 4 content in musculus quadriceps was diminished only in obese females. Terguride treatment lowered prolactin serum levels, which was concomitant with ameliorated insulin sensitivity in obese animals of both sexes. Similarly, only in obese females, terguride significantly increased the affinity of high-affinity insulin-binding sites and normalized GLUT 4 content. Our results document downregulation of insulin receptors and GLUT 4 in obesity and suggest a role for prolactin in obesity-induced insulin resistance, particularly in female rats. PMID- 12079880 TI - Endocrine regulation of subcutaneous fat metabolism during cold exposure in humans. AB - Increased oxidation of carbohydrates and free fatty acids is a well-known phenomenon during cold stress. Nevertheless, sources of the fuels used have not been fully clarified as yet. Thus, the aim of our study was to evaluate the effect of acute cold exposure on lipid and carbohydrate metabolism in human subcutaneous adipose tissue and to identify the possible regulatory mechanisms involved. Ten volunteers were exposed for 30 min to an ambient temperature of 4 degrees C. Interstitial metabolism was assessed with the aid of the microdialysis technique. Lipolysis intensity was evaluated from changes of glycerol concentration in plasma and in dialysate. Cold exposure induced a significant increase of glycerol concentration both in plasma (by 199 +/- 16%, p < 0.01) and in dialysate (by 308 +/- 58%, p < 0.001). No changes in glucose concentration were found whether in plasma or in the dialysate. Ethanol concentration in dialysate increased (148 +/- 15%, p < 0.01), indicating a slower blood flow in the subcutaneous region. Plasma concentrations of various gluco- and/or lipid regulatory hormones remained unaffected by the cold exposure, except for norepinephrine, which rose about threefold (309 +/- 41%, p < 0.001). The data indicate an important role for subcutaneous adipose tissue in mobilization of free fatty acids during cold exposure. This process seems to be regulated by the sympathetic nervous system, whereas hormones involved in the regulation of lipid metabolism, such as epinephrine, insulin, cortisol, and growth hormone, may play a less significant role-at least under the conditions studied. PMID- 12079881 TI - Electrical stimulation improves insulin responses in a human skeletal muscle cell model of hyperglycemia. AB - Myoblasts from human skeletal muscle were isolated from needle biopsy samples of the vastus lateralis of young and healthy volunteers. Contaminating fibroblasts were removed, and myoblasts were fused into differentiated multinucleated myotubes. These myotubes manifested both basal and insulin-stimulated (1-100 nM) glucose transport and glycogen synthesis. Insulin increased 2-deoxyglucose uptake by 1.4-fold and glycogen synthesis by 2.1-fold. Measurements of impedance of cell covered gold electrodes (ECIS system) showed increased micromotion of caffeine stimulated cells, showing their ability to contract. Acute electrical stimulation of the myotubes increased 2-deoxyglucose uptake by about 30%. Treatment with high glucose concentrations (10-20 mM) for 2-8 days reduced both basal and insulin stimulated glucose uptake. Maximal effect was seen after 2 days of treatment with 20 mM glucose. Baseline glucose uptake and glycogen synthesis were reduced by 35%, insulin-stimulated glucose uptake by 25%, and insulin-stimulated glycogen synthesis by 39%. Total cell content of glycogen was not changed by hyperglycemia. The insulin-stimulated glucose uptake in hyperglycemia-treated cells was improved by electrical stimulation of the cells. In conclusion, a model of hyperglycemia has been established, and electrical stimulation improved insulin responses. PMID- 12079882 TI - Differences in oral temperature and body shape in two populations with different propensities for obesity. AB - OBJECTIVE: Body temperature is a function of heat production and heat dissipation. Substantial interindividual variability has been reported in healthy humans. We hypothesized that Pima Indians, a population with a high prevalence of abdominal obesity, may have a lower surface area relative to volume, that is, lower radiating area, and therefore a higher body temperature compared to Caucasians. METHODS: Body composition, including volume (hydrodensitometry), and oral temperature were assessed in 69 nondiabetic Caucasian [age, 30 +/- 7 years; body fat, 21 +/- 8% (mean +/- SD)] and 115 Pima Indian males [age, 27 +/- 6 years; body fat, 28 +/- 6%]. Surface area was estimated from height, weight, and waist circumference (Bouchard's equation). In 47 Pima Indians, measures of insulin sensitivity (M, hyperinsulinemic euglycemic clamp) were available. RESULTS: Compared to Caucasians, Pima Indians had a higher oral temperature [36.4 +/- 0.3 degrees C vs. 36.3 +/- 0.3 degrees C (mean +/- SD), p < 0.04] and lower surface area relative to volume (2.19 +/- 0.05 vs. 2.23 +/- 0.26 m(2), p < 0.0001). Surface area relative to volume was negatively correlated with oral temperature (r = -0.14, p < 0.05), but in a multiple linear regression model it did not entirely explain the ethnic difference in oral temperature. Oral temperature was inversely correlated with M (r = -0.28, p < 0.05). Conclusions Pima Indians have higher oral temperature and lower surface area relative to volume than Caucasians. The ethnic difference in temperature does not seem to be entirely explained by differences in body composition and body shape. Interestingly, higher oral temperature was associated with insulin resistance, a risk factor for type 2 diabetes. PMID- 12079884 TI - Intima-media thickness and atherosclerotic plaques in familial defective apolipoprotein B-100 and familial hypercholesterolemia. AB - Familial defective apolipoprotein B-100 (FDB) is a genetic disorder characterized by a decreased binding of low-density lipoprotein (LDL) particles to the LDL receptor due to defective apo B-100. Impaired LDL clearance could also be due to defects of the LDL receptor (familial hypercholesterolemia, FH). FDB was suggested to be clinically indistinguishable from classical FH. The measurement of the intima-media thickness (IMT) is an accepted method for the direct evaluation of early atherosclerosis. Thus, the aim of this study was to examine the IMT in patients with FDB in comparison to FH. Our data indicate that IMT in FDB does not differ from IMT in FH. PMID- 12079883 TI - Effects of selected anthropometric parameters on plasma lipoproteins, fatty acid composition, and lipoperoxidation. AB - Four anthropometric parameters (body mass index, waist-to-hip circumference ratio, waist circumference, and ratio of subscapularis to triceps skinfold thickness), cutoff values being set according to the WHO guidelines, were compared as discriminative parameters for anthropometric, hemodynamic, and metabolic data relevant as risk factors of coronary heart disease. Waist circumference reflected differences between below and above cutoff values throughout all parameters, with the highest "effect size" values being for fat mass; blood pressure; glucose homeostasis parameters; plasma and LDL cholesterol; plasma, LDL, and VLDL apo B; and LDL oxidability (cumulative effect size value: 2777). Body mass index reflected most significantly plasma, LDL, and VLDL triglyceride; HDL and VLDL cholesterol; and content of linoleic acid in LDL phosphatidylcholine (cumulative effect size value: 2016). Waist-to-hip ratio dominated in effect size value only in VLDL oxidability (cumulative effect size value: 1497). Subscapularis to triceps skinfold thickness ratio had the lowest discriminating ability in all data. PMID- 12079885 TI - Acute elevation of NEFA causes hyperinsulinemia without effect on insulin secretion rate in healthy human subjects. AB - Increased circulating levels of nonesterified free fatty acids (NEFA) have been observed in such hyperinsulinemic states as obesity, impaired glucose tolerance, diabetes, and dyslipidemia where they have been causally linked to the development of insulin resistance and hyperinsulinemia. The concentration of NEFA in plasma is believed to have direct modifying effects on insulin secretion and clearance. It remains controversial whether acute increases in NEFA potentiate insulin secretion in human subjects. We studied the effect of an acute elevation of NEFA during lipid-heparin infusion compared to a glycerol-only control on glucose-stimulated insulin secretion and clearance during a 120-min hyperglycemic (10 mM) clamp in 7 healthy normoglucose-tolerant volunteers. The metabolic clearance rate of C-peptide (MCR(CP)) was measured in each subject during the study by simultaneous infusion of C-peptide. Insulin secretion rate (ISR) was calculated from deconvolution of C-peptide data after correction for the rate of C-peptide infusion. Clearance rate of insulin (MCR(INS)) was calculated based upon endogenous ISR. Plasma glucose (mg/dL): basal (90-115 min) 90.2 +/- 2.8 vs. 90.2 +/- 2.3; clamp (150-240 min) 180.5 +/- 2.8 vs. 180.9 +/- 1.3. Plasma insulin (pmol/L): prebasal (fasting) 29.6 +/- 10.0 vs. 29.8 +/- 10.6; basal (90-115 min) 30.1 +/- 9.2 vs. 34.5 +/- 12.1; second phase clamp (210-240 min) 127.6 +/- 18.2 vs. 182.5 +/- 17.3*. Plasma NEFA (mM): prebasal 0.47 +/- 0.08 vs. 0.52 +/- 0.09; basal 0.35 +/- 0.05 vs. 0.98 +/- 0.02*; clamp (122-240 min) 0.06 +/- 0.02 vs. 0.77 +/- 0.06*. ISR (pmol/min): prebasal 72.7 +/- 7.5 vs. 72.0 +/- 7.9; second phase clamp (210-240 min) 268.5 +/- 27.2 vs. 200.2 +/- 23.7. MCR(INS) (mL/min): prebasal 3393 +/- 488 vs. 3370 +/- 511; clamp 2284 +/- 505 vs. 1214 +/- 153* (*p < 0.05 glycerol vs. intralipid/heparin). This study demonstrates that acute NEFA elevation causes hyperinsulinemia due to a significant decrease in systemic insulin clearance without increasing rates of insulin secretion. PMID- 12079886 TI - Trans fatty acids in subcutaneous fat of pregnant women and in human milk in the Czech Republic. AB - Using capillary gas chromatography, we determined total content of trans fatty acids (TFA) and C18:1 trans fatty acids in human milk and subcutaneous fat in 35 healthy Prague women. The average content of TFA in human milk fat was 4.22% (SD = 1.87%) of all fatty acids, and the value of trans C18:1 isomers was 3.63% (SD = 1.81%). The average concentration of total trans fatty acids in subcutaneous fat was 4.41% (SD = 0.79%) and the average content of C18:1 trans isomers was 2.81% (SD = 0.61%). PMID- 12079887 TI - Detection of a promoter polymorphism in the gene of intestinal fatty acid binding protein (I-FABP). AB - Postprandial fat absorption is supposed to be a major factor in the development of the metabolic syndrome. In recent years, the assimilation of plasma triglycerides has been the focus of several groups, revealing a number of specific fat or fatty acid transporters. The intestinal fatty acid binding protein, I-FABP-2, participates in the absorption of nutritional fats. The influence of a coding polymorphism has been investigated intensively. However, it remains still unclear whether this polymorphism has a major impact on postprandial TG levels in humans. We found a polymorphism in the promoter of FABP 2, which might involve the retinoid receptor in the transcriptional activity. In functional analysis, we have been able to demonstrate that the various promoter alleles develop different activities in the human intestinal epithelial cells and that the postprandial appearance of plasma TGs in healthy subjects also depends on their genotype. Since the distribution of the identified promoter polymorphism does not differ in subjects suffering from type 2 diabetes, the overall influence on the development of the metabolic syndrome seems to be minor. PMID- 12079888 TI - Glucose transporter 4 gene: association studies pertaining to alleles of two polymorphisms in extremely obese children and adolescents and in normal and underweight controls. AB - The human insulin-responsive glucose transporter 4 gene (GLUT4) has been related to non-insulin-dependent diabetes mellitus (NIDDM) in several studies. Obesity is commonly found in patients with NIDDM. Hence, genes involved in NIDDM might also be relevant for obesity. We have analyzed 212 extremely obese children and adolescents, 82 normal-weight students, and 94 underweight students for two single nucleotide polymorphisms (SNPs: promoter -30G/A; exon 4a: silent 2061T/C) in the vicinity of the GLUT4 by polymerase chain reaction with subsequent restriction fragment length polymorphism analyses (PCR-RFLP) or single-strand conformation polymorphism analyses (SSCP). Allele and genotype distributions were similar in all study groups (all p values > 0.05). Hence, we did not detect association of any of the analyzed SNP alleles in the GLUT4 to different weight extremes, so these seem not to be involved in weight regulation in our study groups. PMID- 12079889 TI - Association of insulin gene VNTR polymorphism with polycystic ovary syndrome. AB - Variability in the number of tandem repeats of the insulin gene (INS VNTR) is known to influence several phenotypes, including polycystic ovary syndrome (PCOS), diabetes mellitus type 1, diabetes mellitus type 2, and birth weight. The presence of the class III allele of INS VNTR has been reported to be protective in diabetes mellitus type 1, but in contrary it increases the disease risk of PCOS and diabetes mellitus type 2. PCOS is a very common endocrinopathy in women of reproductive age. The etiology of PCOS is uncertain, but family history of this syndrome suggests a major genetic cause. The aim of this pilot study was to investigate the possible association of INS VNTR polymorphism with PCOS in Czech women. In PCOS, significantly higher WHR, BMI, G(0), G(180), I(30), Cp(0), Cp(30), Cp(60), AUC-I, AUC-Cp, and insulinogenic index and significantly lower AUC-G/AUC-I were found. No significant differences in INS VNTR genotype, phenotype, or allele frequencies were found between PCOS and controls. In spite of several differences in anthropometric and biochemical parameters (abdominal fat localization, increased beta-cell function, and lower insulin sensitivity in PCOS women), no effect of INS VNTR polymorphism was found on insulin secretion, insulin action, or any other screened parameter. PMID- 12079891 TI - Increased abdominal obesity in subjects with a mutation in the 5-HT(2A) receptor gene promoter. AB - In the present study, we examined the potential impact of the 5-HT(2A) -1438G/A promoter polymorphism on obesity and estimates of insulin, glucose, and lipid metabolism as well as circulating hormones, including salivary cortisol, in 284 unrelated Swedish men born in 1944. The subjects were genotyped by using PCR amplification of the promoter region of the gene for 5-HT(2A) followed by digestion with the restriction enzyme MspI. The frequencies were 0.39 for allele 1438A and 0.61 for allele -1438G. Homozygotes for the -1438G allele had, in comparison with -1438A/A subjects, higher body mass index (BMI), waist-to-hip ratio (WHR), and abdominal sagittal diameter. Moreover, cortisol escape from 0.25 mg dexamethasone suppression was found in subjects with the -1438A/G genotype. Serum leptin, fasting insulin and glucose, as well as serum lipids were not different across the -1438G/A genotype groups. From these results, we suggest the possibility that an abnormal production rate of the 5-HT(2A) gene product might lead to the development of abdominal obesity. The pathophysiology could involve stress factors that destabilize the serotonin-hypothalamic-pituitary-adrenal systems in those with genetic vulnerability in the serotonin receptor gene. PMID- 12079890 TI - Association between a variant at the GABA(A)alpha6 receptor subunit gene, abdominal obesity, and cortisol secretion. AB - In the present study, we examined the potential impact of a T-to-C substitution at nucleotide 1519 of the GABA(A)alpha6 receptor subunit gene (GABRA6) on obesity and obesity-related phenotypes as well as circulating hormones, including salivary cortisol, in 284 unrelated Swedish men born in 1944. The subjects were genotyped by using PCR amplification followed by digestion with the restriction enzyme AlwNI. The frequency of allele T was 0.54 and that of allele C was 0.46. Carriers for the T allele (n = 211) had higher waist-to-hip ratio (p = 0.094) and abdominal sagittal diameter (p = 0.084) compared to homozygotes for the C allele (n = 56). The homozygotes for the T allele had, in comparison to heterozygotes, significantly (p = 0.004-0.024) higher mean cortisol levels at 11:45 am; at 30, 45, and 60 min after a standardized lunch; and finally at 5:00 pm. In addition, T/T subjects had significantly (p = 0.031) higher diurnal cortisol secretion compared to T/C subjects. Leptin, insulin, and glucose were not different across the genotype groups. In conclusion, these findings suggest a role of the point substitution (T-to-C) at nucleotide 1519 of GABRA6 in the predisposition to hypercortisolism and perhaps abdominal obesity. The pathophysiology may involve various environmental factors, particularly stress, that destabilize the GABA hypothalamic-pituitary-adrenal systems in those with genetic vulnerability. PMID- 12079892 TI - Lipids and insulin resistance: what we've learned at the Fourth International Smolenice Symposium. AB - A summary of the Fourth International Smolenice Symposium on Lipids and Insulin Resistance focusing on "The Role of Fatty Acid Metabolism and Fuel Partitioning" is provided. Highlights and issues of the conference are mentioned, as well as strategies for the future. PMID- 12079893 TI - Vaccination and rheumatoid arthritis. PMID- 12079894 TI - Anti-interleukin 1alpha autoantibodies. PMID- 12079896 TI - Shortness of breath in systemic lupus erythematosus: a diagnostic and therapeutic dilemma. PMID- 12079897 TI - Case number 24: scalp necrosis in giant cell arteritis. PMID- 12079895 TI - Reactive arthritis or chronic infectious arthritis? AB - Microbes reach the synovial cavity either directly during bacteraemia or by transport within lymphoid cells or monocytes. This may stimulate the immune system excessively, triggering arthritis. Some forms of ReA correspond to slow infectious arthritis due to the persistence of microbes and some to an infection triggered arthritis linked to an extra-articular site of infection. PMID- 12079899 TI - Autoantibodies against interleukin 1alpha in rheumatoid arthritis: association with long term radiographic outcome. AB - OBJECTIVES: To investigate the possible association of interleukin 1alpha autoantibodies (IL1alpha aAb) with the long term course of joint erosion in patients with rheumatoid arthritis (RA). METHODS: Serum samples from 176 patients with RA included in a prospective study over 30 years were analysed for IL1alpha aAb by binding to human [(125)I]IL1alpha. Erosions of 19 diarthrodial joints were radiographically scored by the Larsen method. RESULTS: The relative risk (RR) of early IL1alpha aAb positive patients developing at least 30% of maximum radiographic joint destruction was significantly lower than for IL1alpha aAb negative patients, RR=0.29 (p=0.04). In rheumatoid factor positive patients RR was only 0.18 (p=0.02). Patients who seroconverted more than two years after the onset of RA showed the most aggressive development of joint erosion, with a relative risk of at least 40% of maximum radiographic joint destruction of 2.56 (p=0.048) CONCLUSIONS: The progression of radiographic joint destruction in patients with RA is associated with, and perhaps modified by, circulating IL1alpha aAb, suggesting that IL1alpha or IL1alpha aAb, or both, have a role in the erosive processes. IL1alpha aAb appear to be of prognostic significance in RA. PMID- 12079900 TI - Rheumatoid factor measured by fluoroimmunoassay: a responsive measure of rheumatoid arthritis disease activity that is associated with joint damage. AB - OBJECTIVES: To determine whether rheumatoid factors (RFs), measured as continuous variables by time resolved fluoroimmunoassay, reflect disease activity in rheumatoid arthritis (RA). Further, to study the association of RFs and other disease activity parameters with radiological joint damage, especially in individual patients. METHODS: In active, early RA, IgM and IgA RFs, as well as erythrocyte sedimentation rate (ESR), C reactive protein (CRP), tender joint score, and swollen joint score were assessed regularly. At the study start and at 56 and 80 weeks, radiographs of hands and feet were assessed by the Sharp score (van der Heijde modification). Associations between RFs and disease activity parameters were studied. In addition, associations between radiographic damage and disease activity parameters (baseline and time integrated) were analysed by non-parametric tests and multiple regression analysis. The relation between time integrated disease activity parameters and radiological damage in individual patients was analysed and visualised. RESULTS: 155 patients were included. RF levels were strongly associated with the disease activity parameters (especially ESR and CRP) and with each other. All disease activity parameters, at baseline as well as time integrated parameters, were associated with (the progression of) radiographic damage. Moreover, in individual patients, a linear relationship between time integrated disease activity parameters and progression of radiological damage was seen. CONCLUSION: RFs, measured as continuous variables, can be considered as disease activity parameters in patients with RA. The level of RF at baseline and the exposure to RF over time is associated with radiological damage. In individual patients, there is a constant relation between disease activity and radiological damage. PMID- 12079898 TI - Differential expression patterns of matrix metalloproteinases and their inhibitors during development of osteoarthritis in a transgenic mouse model. AB - OBJECTIVE: To characterise the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) during degeneration of articular cartilage in a transgenic Del1 mouse model for osteoarthritis. METHODS: Northern analysis was used to measure mRNA levels of MMP-2, -3, -8, -9, -13, and -14, and TIMP-1, -2, and -3 in total RNA extracted from knee joints of transgenic Del1 mice, harbouring a 15 amino acid deletion in the triple helical domain of the alpha1(II) collagen chain, using their non-transgenic littermates as controls. Immunohistochemistry was used to study the presence of cleavage products (neoepitopes) of type II collagen, and the distribution of MMP-13 and TIMP-1 in degenerating cartilage. RESULTS: Each of the MMP and TIMP mRNAs analysed exhibited distinct expression patterns during development and osteoarthritic degeneration of the knee joint. The most striking change was up regulation of MMP 13 mRNA expression in the knee joints of Del1 mice at the onset of cartilage degeneration. However, the strongest immunostaining for MMP-13 and its inhibitor TIMP-1 was not seen in the degenerating articular cartilage but in synovial tissue, deep calcified cartilage, and subchondral bone. The localisation of type II collagen neoepitopes in chondrocytes and their pericellular matrix followed a similar pattern; they were not seen in cartilage fibrillations, but in adjacent unaffected cartilage. CONCLUSION: The primary localisation of MMP-13 and TIMP-1 in hyperplastic synovial tissue, subchondral bone, and calcified cartilage suggests that up regulation of MMP-13 expression during early degeneration of articular cartilage is a secondary response to cartilage erosion. This interpretation is supported by the distribution of type II collagen neoepitopes. Synovial production of MMP-13 may be related to removal of tissue debris released from articular cartilage. In the deep calcified cartilage and adjacent subchondral bone, MMP-13 probably participates in tissue remodelling. PMID- 12079902 TI - How reliably do rheumatologists measure shoulder movement? AB - OBJECTIVE: To assess the intrarater and interrater reliability among rheumatologists of a standardised protocol for measurement of shoulder movements using a gravity inclinometer. METHODS: After instruction, six rheumatologists independently assessed eight movements of the shoulder, including total and glenohumeral flexion, total and glenohumeral abduction, external rotation in neutral and in abduction, internal rotation in abduction and hand behind back, in random order in six patients with shoulder pain and stiffness according to a 6x6 Latin square design using a standardised protocol. These assessments were then repeated. Analysis of variance was used to partition total variability into components of variance in order to calculate intraclass correlation coefficients (ICCs). RESULTS: The intrarater and interrater reliability of different shoulder movements varied widely. The movement of hand behind back and total shoulder flexion yielded the highest ICC scores for both intrarater reliability (0.91 and 0.83, respectively) and interrater reliability (0.80 and 0.72, respectively). Low ICC scores were found for the movements of glenohumeral abduction, external rotation in abduction, and internal rotation in abduction (intrarater ICCs 0.35, 0.43, and 0.32, respectively), and external rotation in neutral, external rotation in abduction, and internal rotation in abduction (interrater ICCs 0.29, 0.11, and 0.06, respectively). CONCLUSIONS: The measurement of shoulder movements using a standardised protocol by rheumatologists produced variable intrarater and interrater reliability. Reasonable reliability was obtained only for the movement of hand behind back and total shoulder flexion. PMID- 12079901 TI - Prevalence and clinical significance of anti-cyclic citrullinated peptide antibodies in juvenile idiopathic arthritis. AB - BACKGROUND: Antibodies against cyclic citrullinated peptide (anti-CCP) are considered to be specific for rheumatoid arthritis (RA). OBJECTIVE: To assess the clinical significance of anti-CCP in a cohort of patients with juvenile idiopathic arthritis (JIA). METHODS: Anti-CCP were tested by an enzyme linked immunosorbent assay (ELISA) in serum samples from 109 patients with JIA (30 boys, 79 girls), with a mean age of 8.7 years (range 0.6-20.3) and mean disease duration of 3.6 years (range 3 months to 15.6 years). As control groups, anti-CCP were also tested in sera of 30 healthy children, 25 patients with juvenile onset systemic lupus erythematosus (SLE), and 50 adult patients (30 with RA, 20 with SLE). RESULTS: Positive anti-CCP values were found in sera of two patients with JIA (2%), one with polyarthritis, and one with oligoarthritis. Statistical analysis showed that anti-CCP were not associated with the presence of antinuclear antibodies, raised erythrocyte sedimentation rate, or erosions. In the control groups, none of the patients with juvenile onset SLE and only one of 20 adults with SLE were positive for anti-CCP, but 19/30 (63%) adults with RA showed anti-CCP positivity. CONCLUSIONS: Anti-CCP can be detected in children with JIA, but are less frequently present than in adults with RA. PMID- 12079903 TI - Dynamic load at baseline can predict radiographic disease progression in medial compartment knee osteoarthritis. AB - OBJECTIVE: To test the hypothesis that dynamic load at baseline can predict radiographic disease progression in patients with medial compartment knee osteoarthritis (OA). METHODS: During 1991-93 baseline data were collected by assessment of pain, radiography, and gait analysis in 106 patients referred to hospital with medial compartment knee OA. At the six year follow up, 74 patients were again examined to assess radiographic changes. Radiographic disease progression was defined as more than one grade narrowing of minimum joint space of the medial compartment. RESULTS: In the 32 patients showing disease progression, pain was more severe and adduction moment was higher at baseline than in those without disease progression (n=42). Joint space narrowing of the medial compartment during the six year period correlated significantly with the adduction moment at entry. Adduction moment correlated significantly with mechanical axis (varus alignment) and negatively with joint space width and pain score. Logistic regression analysis showed that the risk of progression of knee OA increased 6.46 times with a 1% increase in adduction moment. CONCLUSIONS: The results suggest that the baseline adduction moment of the knee, which reflects the dynamic load on the medial compartment, can predict radiographic OA progression at the six year follow up in patients with medial compartment knee OA. PMID- 12079904 TI - Safety and efficacy of vaccination against hepatitis B in patients with rheumatoid arthritis. AB - BACKGROUND: Hepatitis B infection and vaccination against it have been implicated in the potential triggering or flare of some autoimmune diseases, including rheumatoid arthritis (RA). However, the safety of hepatitis B vaccination in patients with pre-existing RA is not known. OBJECTIVES: To assess the safety and antibody response of immunisation with a recombinant DNA hepatitis B vaccine in patients with RA. PATIENTS AND METHODS: The study comprised 44 patients with RA, of whom 22 received three doses (the second and third dose being given after one and six months) of a recombinant DNA hepatitis B vaccine (study group) and 22 did not receive the vaccine (control group). Both groups had comparable proportions of women and similar mean age (51 years). Clinical assessment before and two and seven months after the first immunisation included evaluation of daytime pain with a 10 cm visual analogue scale, duration of morning stiffness, and number of tender and swollen joints. Erythrocyte sedimentation rate (ESR) and C reactive protein (CRP) were measured at each visit. Antibodies to hepatitis B surface antigen (HBsAg) were determined by a commercial enzyme linked immunosorbent assay (ELISA) test kit. RESULTS: Hepatitis B vaccination was not associated with an appreciable deterioration in any clinical or laboratory measure of disease. The measures of disease activity of the patients and controls during the study period did not differ significantly: p=0.76 for daytime pain, p=0.1 for morning stiffness, p=0.24 and p=0.3 for tender and swollen joints respectively, p=0.08 for CRP, and p=0.12 for ESR. Fifteen of the 22 patients responded to vaccination, with an antibody level against HBsAg of 10 IU/l after seven months. Lack of response was associated with older age and higher scores of daytime pain. CONCLUSIONS: Hepatitis B vaccination is safe in RA and produces antibodies in 68% of the patients. PMID- 12079905 TI - Laboratory and imaging studies used by French rheumatologists to determine the cause of recent onset polyarthritis without extra-articular manifestations. AB - BACKGROUND: The cause of recent onset polyarthritis can be difficult to identify. OBJECTIVE: To determine which laboratory and imaging studies French rheumatologists recommend, not taking cost into account, for the diagnosis of recent onset polyarthritis without extra-articular manifestations. METHODS: From the list of the French Society for Rheumatology, a random sample of 210 rheumatologists was selected, who were asked to complete a questionnaire on the laboratory and imaging studies they would recommend in two fictional cases of recent onset polyarthritis (possible rheumatoid arthritis (RA)-case 1 and probable RA-case 2). RESULTS: In case 1, the following were recommended by over 75% of respondents: hand radiographs, rheumatoid factors (RFs), and antinuclear antibodies (ANA) (92%, 98%, and 98%, respectively). 50-74% of respondents recommended radiographs of the feet, knees, and chest (50%, 57%, and 66%, respectively); blood cell counts, erythrocyte sedimentation rate (ESR), serum assays of C reactive protein (CRP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) (65%, 74%, 67%, and 62%, respectively). 25-49% recommended determination of creatinine and proteinuria, HLA-B27, antikeratin antibody, radiographs of the pelvis, and synovial fluid analysis. Several investigations were recommended less often in case 2 than in case 1. Nevertheless, some laboratory and imaging studies (radiographs of hand, feet, knees, chest x rays, blood cell counts, ANA, RF, antikeratin antibody, CRP, ESR, creatinine, AST and ALT, proteinuria, and joint aspiration) were recommended by more than 25% of respondents in both cases. CONCLUSION: Wide variations were found among rheumatologists, indicating a need for standardisation. Some laboratory and imaging studies are recommended by at least 25% of respondents in recent onset polyarthritis with or without clues suggesting RA. In contrast, many tests were considered useful by fewer than 25% of the respondents in both cases. PMID- 12079906 TI - Practical progress in realisation of early diagnosis and treatment of patients with suspected rheumatoid arthritis: results from two matched questionnaires within three years. AB - BACKGROUND: Early diagnosis and treatment with disease modifying antirheumatic drugs (DMARDs) have been advocated for patients with rheumatoid arthritis (RA). This survey focuses on the individual definitions and treatment modalities of rheumatologists, and aims at determining the practical realisation of these concepts. METHODS: A questionnaire to be self completed was handed out at the EULAR Symposium 1997. The main issues dealt with were definition, referral time, diagnosis, follow up, and treatment of early RA. Of the 111 participants, who were from all continents and all age groups, 85 (77%) gave their name and address. In 2000, the same questionnaire was sent to these 85 primary respondents. Forty four questionnaires (52%) were returned, and their results were matched and further evaluated. RESULTS: The definition of early RA was heterogeneous, but two of three rheumatologists use the term "early" for symptoms shorter than three months. There was a drift towards acceptance of involvement of fewer affected joints. Serological tests obtained for early diagnosis were mostly rheumatoid factor and antinuclear antibodies, usually in combination (approximately 70%), while other tests (antikeratin antibodies, antiperinuclear factor, anti-RA33) were used rarely, but increasingly (21-25% all together). No significant change in the lag time of referral to the specialist of patients with suspected early RA was seen within these three years (<3 months for 50%, >6 months for 20%), while the proportion followed up during the first three months increased. At both times, every second rheumatologist started DMARD treatment only when the 1987 American College of Rheumatology (ACR) criteria were fulfilled. However, in 1997 about 10% were willing to wait for erosions before starting DMARDs, while none did so in 2000. Methotrexate, sulfasalazine, and antimalarial drugs were the most commonly prescribed DMARDs in early RA, with the first two of these still being in increasing use. CONCLUSION: The understanding of "early" rheumatoid arthritis is heterogeneous, but the vast majority of the rheumatologists surveyed regard symptom duration of <3 months as early. Rheumatoid factor was the most useful laboratory support in early diagnosis. Because there has been no shortening of referral time of patients with new RA within the past three years, and many rheumatologists start DMARDs only when the ACR criteria are fulfilled, it is concluded that guidelines for early referral, as well as for early (rheumatoid) arthritis, are needed. PMID- 12079907 TI - Prevalence of hepatitis C virus infection in patients with rheumatoid arthritis. AB - BACKGROUND: Various viruses have been implicated in the cause and pathogenesis of rheumatoid arthritis (RA). Hepatitis C virus (HCV) infection, which has been recognised as a cause of some autoimmune diseases, and which has been described as sometimes presenting with rheumatic manifestations indistinguishable from RA, might be a candidate. OBJECTIVE: To evaluate the prevalence of HCV infection in patients with RA. METHODS: Consecutive patients with RA admitted to hospital in two departments of rheumatology were prospectively studied. Patients' serum samples were screened for the presence of anti-HCV antibodies. Patients with positive serology were further evaluated for the presence of HCV ribonucleic acid by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: 309 patients (232 women, 77 men, mean age (SD) 54.1 (14.8) years) were studied. Their mean (SD) disease duration was 74.1 (91) months. Tests for rheumatoid factors and antinuclear antibodies were positive in 213 (69%) and 114 (37%) of the patients respectively. Systemic vasculitis was found in 12 (4%) of the patients. Mean erythrocyte sedimentation rate was 36.4 (SD 30.5) mm at the first hour (normal <10 mm) and C reactive protein was 36.8 (SD 45.8) mg/l (normal range <5 mg/l), respectively, with 181(58.6%) of patients considered as having active disease. Aspartate transaminases were increased in 14 (4%) patients, and alkaline phosphatase in 14 (4%). A positive anti-HCV serology was found in two (0.65%) patients, including one with a previously diagnosed HCV infection. HCV RNA was positive by RT-PCR in one of those two patients. CONCLUSION: A 0.65% prevalence of past or active HCV infection was found in patients with RA, which did not differ from the prevalence of HCV in the general French population. This result does not support the participation of HCV infection in the pathogenesis of RA. PMID- 12079908 TI - Decreased flares of rheumatoid arthritis during the first year of etanercept treatment: further evidence of clinical effectiveness in the "real world". AB - OBJECTIVE: To determine the incidence of disease flare during the first year of etanercept treatment for 88 patients with rheumatoid arthritis (RA) and compare it with the incidence of flare in those same patients in the year before etanercept use. METHODS: The outpatient clinic charts of all patients with RA who were prescribed etanercept in or before September 1999, who also had at least one year's follow up in the same outpatient clinic, were surveyed. The primary outcome measure was the number of disease flares in one year before and after etanercept use. The secondary outcome measures included the number of patients who did and did not flare, how flares were treated, and the drug alterations that were necessary during the same two time intervals. RESULTS: The total number of flares for all patients in the year before etanercept treatment was 214 (mean (SD) 2.43 (1.75)). The number of flares in the first year of etanercept treatment decreased to 83 (mean 0.94 (1.07)) (p<0.0001). The total number of patients who had at least one flare in the year before etanercept use was 80; eight had no flares. In their first year of etanercept treatment, 50 patients had at least one flare; 38 had no flares (p<0.0001). Twenty one patients (24%) stopped using etanercept before completing one year's treatment. CONCLUSION: This study of patients with RA in the "real world" shows that etanercept is effective in reducing the number of RA flares. PMID- 12079909 TI - Helicobacter pylori does not play a part in the dyspeptic complaints of rheumatology patients receiving long term treatment with non-steroidal anti inflammatory drugs. AB - BACKGROUND: The presence of dyspeptic symptoms is a common finding in patients treated with non-steroidal anti-inflammatory drugs (NSAIDs). Some studies seem to support the involvement of Helicobacter pylori infection in the dyspeptic symptoms reported by these patients, and suggest that eradication may be useful. OBJECTIVE: To determine the variables related to dyspepsia in rheumatology patients requiring NSAID treatment, assessing in particular the role of Helicobacter pylori infection. METHODS: One hundred and eighty six consecutive patients with a rheumatological disorder requiring NSAID treatment (68 male, 118 female; mean (SD) age 55 (15) years) were included in a cross sectional study; dyspeptic symptoms were measured by a previously validated scale. Helicobacter pylori infection was determined by serology. Variables related to the severity of symptoms and the need for antisecretory drugs were determined by multivariate analysis. RESULTS: No relation was found between Helicobacter pylori infection and dyspepsia or any of its surrogate markers (antisecretory drug use or NSAID intolerance). Female sex and treatment with antisecretory drugs were found to be independent predictors for the appearance and severity of dyspeptic symptoms. The only independent predictive variables of the requirement for antisecretory drugs were age, previous ulcer disease, taking NSAIDs with a medium or high anti inflammatory potential, and the symptoms score. CONCLUSION: Helicobacter pylori infection does not seem to play any part in the gastric symptoms of patients treated long term with NSAIDs. PMID- 12079910 TI - A six year prospective study of a cohort of patients with fibromyalgia. AB - OBJECTIVES: To examine the long term prognosis in patients with fibromyalgia (FM). METHODS: Forty five of 70 patients who had participated in a three week trial six years earlier completed again the same questionnaires used previously. RESULTS: Most symptoms had remained stable. Pain had increased, but some aspects of quality of life had improved over time. CONCLUSION: Symptoms of FM persisted over the six years, but patients appeared better able to cope with them. PMID- 12079911 TI - Diagnosing patients with longstanding shoulder joint pain. AB - OBJECTIVE: To examine the interobserver agreement of commonly used clinical tests and diagnoses in patients with shoulder pain, and the accuracy of these tests and ultrasonographic findings in comparison with arthroscopic findings. METHODS: Eighty six patients with longstanding shoulder joint pain were "blindly" examined by two trained doctors using several clinical tests. In all patients an ultrasonographic examination was performed, and in 42 (49%) an arthroscopy. RESULTS: Tests for impingement showed poor to moderate agreement. Tenderness of muscles, muscle weakness, and tests for labral lesion also showed poor agreement. Pain during muscle contraction showed moderate agreement. The agreement of clinical diagnoses was poor and the accuracy was low in comparison with arthroscopy. Ultrasonography was accurate in full thickness supraspinatus tendon tears, but inaccurate for partial tears and labral lesions. CONCLUSIONS: Most clinical tests showed poor agreement. Clinical and ultrasonographic diagnoses had low accuracy in comparison with arthroscopy. PMID- 12079912 TI - The cervical spine in patients with psoriatic arthritis. AB - OBJECTIVE: To establish the incidence of clinically important inflammatory cervical spine abnormalities in radiographs of patients with psoriatic arthritis (PsA). METHODS: Patients were selected from a rheumatological outpatient clinic and one ward of the Rheumatism Foundation Hospital, Heinola, Finland, by examining 160 consecutive PsA cases. A total of 65 patients (38 women, 27 men) with PsA were identified who had cervical spine radiographs available. These were evaluated for inflammatory changes, and patient records studied for disease characteristics, laboratory and clinical findings. RESULTS: In 12 cases (18%) inflammatory cervical spine changes were seen in the cervical spine radiographs. The most frequently detected was apophysial joint ankylosis, seen in seven patients (11%). Anterior atlantoaxial subluxation (aAAS) was seen in five (8%) and atlantoaxial impaction in three (5%). In 20 of the 40 patients who had the rotational range of neck motion measured the measurement was < or =45 degrees either to the left or the right side. CONCLUSION: Inflammatory cervical spine changes were not commonly seen in radiographs of patients with PsA. Apophysial joint ankylosis and aAAS were detected most often. PsA may decrease the rotational range of neck motion significantly. PMID- 12079913 TI - Peptostreptococcal pericarditis complicating anti-tumour necrosis factor alpha treatment in rheumatoid arthritis. PMID- 12079915 TI - Paediatric Behcet's disease in France. PMID- 12079914 TI - Alopecia in Wegener's granulomatosis. PMID- 12079916 TI - Intra-articular and soft tissue injections: assessment of the service provided by nurses. PMID- 12079917 TI - Manual jobs increase the risk of patients with ankylosing spondylitis withdrawing from the labour force, also when adjusted for job related withdrawal in the general population. PMID- 12079918 TI - Breast implants and illness: a model of psychological illness. PMID- 12079919 TI - Fibromyalgia in patients with rheumatoid arthritis is associated with higher scores of disability. PMID- 12079920 TI - Shingles following infliximab infusion. PMID- 12079921 TI - Self limiting lupus-like symptoms in patients with parvovirus B19 infection. PMID- 12079922 TI - Dermatomyositis in a patient with adenocarcinoma of the gall bladder. PMID- 12079923 TI - Coincidence of asymptomatic avascular necrosis and fracture of the femoral neck: a rare combination of glucocorticoid induced side effects. PMID- 12079924 TI - Usefulness of bone densitometry in postmenopausal women with clinically diagnosed vertebral fractures. PMID- 12079925 TI - Soluble TNF receptor treatment does not affect raised TGF beta levels in RA. PMID- 12079926 TI - Infection and SLE. PMID- 12079927 TI - Was it a case of Takayasu arteritis? PMID- 12079928 TI - Forty years of homograft surgery. PMID- 12079929 TI - On-pump versus off-pump coronary artery bypass using quadruple arterial grafts. AB - The optimal combination of arterial grafts for quadruple coronary artery bypass is the bilateral internal mammary artery, the radial artery, and the gastroepiploic artery. Patients who underwent quadruple bypass in our hospital group between December 1995 and March 2001 were retrospectively analyzed to determine whether off-pump surgery (n = 27) provides better outcomes than conventional on-pump surgery (n = 51). Preoperative risk factors as well as the mean number of distal anastomoses (4.5 in the off-pump versus 4.8 in the on-pump patients) were not significantly different between the 2 groups. Postoperative recovery was significantly faster in the off-pump group than in the on-pump group (intubation time, 6.4 versus 16 hours; stay in intensive care unit, 2 versus 3 days; and postoperative hospital stay, 12.3 versus 15.8 days). Early stenosis free graft patency rates did not differ significantly (90.3% versus 89.3%). No late cardiac events were observed in the off-pump group, while 4 occurred in the on-pump group. Quadruple arterial bypass without cardiopulmonary bypass is safe, and it allows faster recovery. The follow-up results of off-pump patients so far have been satisfactory. PMID- 12079930 TI - Coronary bypass surgery in patients on thyroxin replacement therapy. AB - The outcome of coronary bypass surgery was analyzed in 25 patients who were on thyroxin replacement therapy for chronic thyroid disorders at the time of operation. It was hypothesized that if such patients were given only their routine dose of thyroxin on the day of surgery, hemodynamic and cardiorespiratory recovery may be poor. All the patients on thyroxin replacement therapy were given their routine dose of thyroxin orally or via a nasogastric tube in the perioperative period. No supplemental dose was used. Based on preoperative levels of thyroid stimulating hormone, 68% of these patients were biochemically hypothyroid prior to surgery. Analysis of a large number of variables showed no difference in outcome against a control group who had no previous thyroid problems. We conclude that routine thyroxin administration is all that is required for a satisfactory outcome in patients undergoing coronary bypass surgery while on thyroxin replacement therapy. PMID- 12079931 TI - Clinical features and surgical treatment of cardiac myxoma: report of 18 cases. AB - Myxomas are the most common benign primary cardiac tumors. As many patients with cardiac myxoma suffer from cerebral or systemic embolism, which are serious complications, diagnosis of the tumor is vital. Between 1990 and 2000, 18 patients (6 males, 12 females), aged 24 to 73 years (mean, 55.3 years), were operated on for cardiac myxoma. The most common location of the myxoma was the left atrium (78%), and the transseptal surgical approach was preferred (78%) as it allows total resection of the left atrial myxoma along with its pedicle. Carney complex, a familial autosomal dominant form of atrial myxoma, was not found in any of the patients. There had been no operative or postoperative mortality and morbidity. The mean postoperative follow-up period was 5 years (mean, 1 to 10 years). No recurrence had been seen. We believe that the transseptal approach, in allowing total resection of the myxoma, prevents recurrence. PMID- 12079932 TI - Is hyperamylasemia after cardiac surgery due to cardiopulmonary bypass? AB - Although hyperamylasemia has been reported in a large proportion of patients undergoing cardiac surgery with cardiopulmonary bypass, its clinical significance and pathogenetic mechanisms remain poorly understood. The study was designed to investigate whether avoidance of cardiopulmonary bypass would limit amylase elevation. Serum levels of amylase and lipase were measured preoperatively as well as 24 and 48 hours postoperatively in 58 patients undergoing elective coronary artery bypass grafting. Three surgical approaches were used: cardiopulmonary bypass (n = 32) and off-pump through a median sternotomy (n = 14) or a left minithoracotomy (n = 12). There was no hospital mortality or postoperative abdominal complications. Transient hyperamylasemia occurred in 14 patients: 7 (22%), 5 (36%), and 2 (17%) in the respective groups. The increase in amylase levels was similar among the groups. However, no lipase elevation was detected in any patient. There was no clear correlation between hyperamylasemia and increased creatinine levels. Perioperative plasma calcium levels were normal in patients who had hyperamylasemia. Our results indicate that hyperamylasemia after bypass surgery is not related to the use of cardiopulmonary bypass or the mode of surgical access. PMID- 12079933 TI - Clinical effects of different protamine doses after cardiopulmonary bypass. AB - The optimal dose of protamine needed to reverse the anticoagulant effect of heparin after cardiopulmonary bypass is still not known. In this retrospective cohort study, we investigated 3 different dose regimes in 300 patients undergoing coronary artery bypass grafting. Group A patients (n = 100) were given protamine in the ratio of 1.3 mg to 1 mg heparin, group B patients (n = 100) were given 0.75 mg protamine to 1 mg heparin, and group C patients (n = 100) were given protamine in fractionated doses of 1 mg + 0.15 mg + 0.15 mg to 1 mg heparin. The groups were comparable in all major clinical and operative variables. The heparin dose was almost identical in the groups. The rate of red cell transfusion was significantly higher in group B than in the other groups. A similar but nonsignificant trend was observed in the incidence of resternotomy for postoperative bleeding, mediastinal drainage, and postoperative hemoglobin loss. The study demonstrates that a single bolus dose of 1.3 mg protamine to 1 mg heparin is safe and efficient for neutralizing heparin after cardiopulmonary bypass. PMID- 12079934 TI - Arterial duct closure with detachable coils: application in the small child. AB - Transcatheter closure of patent ductus arteriosus using controlled-release coils was performed in 16 patients weighing < 10 kg. No embolization occurred. Procedure-related complications occurred in 3 patients (18.8%): massive femoral hemorrhage in 1 and femoral artery thrombosis in 2. The ductus recanalized in 1 of them because of mechanical hemolysis caused by streptokinase treatment. This was the only patient who underwent another occlusion procedure. Complete occlusion was achieved in 7 patients (43.8%) immediately, in 13 (81.2%) the following day, and in all 15 patients who had completed the 6-month follow-up. During follow-up, flow velocities between the left and the main pulmonary arteries and between the descending and the ascending aortae did not differ significantly. Flow velocity was > 2 m x sec(-1) in 3 patients in the left pulmonary artery and in 1 in the descending aorta. Protrusion of the coil was seen in 3 of these patients. Flow velocity was also high in the main pulmonary artery in the 4th patient. In conclusion, coil occlusion of ductus arteriosus is feasible in the small child, but no more than half a loop of the coil should be left at the pulmonary site. High flow velocity does not always mean obstruction. PMID- 12079935 TI - Arterial complications following cardiac catheterization in children less than 10 kg. AB - We sought to determine if a higher dose of heparin would reduce arterial complications in patients weighing 10 kg or less undergoing cardiac catheterization to investigate congenital heart disease. Sixty patients were given either 100 (group A) or 150 (group B) IU x kg(-1) of heparin in a double blinded randomized manner. Initial arterial access was established using a 4F cannula in all patients. Mean activated clotting time measured 20 minutes following heparin administration was significantly lower in group A than in group B (199 versus 251 seconds). Only 3 out of 60 patients (5%) required treatment for loss of femoral pulse. The age, weight, activated clotting time, length of catheterization procedure, time taken to establish arterial access, and the duration of arterial cannulation were comparable between the groups. Weight under 4 kg, age under 1 month, and cannula size larger than 4F were identified as independent risk factors for the development of arterial complications. Arterial access using a 4F cannula is a safe procedure in children weighing 10 kg or less. The incidence of significant arterial complications is low, and they do not appear to be preventable by a higher dose of heparin. PMID- 12079936 TI - Robotically controlled video-assisted port-access mitral valve surgery. AB - From 1997 to 2000, 221 patients underwent mitral valve surgery through a mini thoracotomy, using a port-access endovascular cardiopulmonary bypass system in 38 and a transthoracic clamp in 183. In 120 patients, exposure of the mitral valve was facilitated by an endoscope attached to a voice-controlled robotic arm (AESOP 3000). The mitral valve was repaired in 26 patients and replaced in 195; 24 were redo cases. Operating time was 3.5 +/- 1.2 hours, aortic crossclamp time was 58 +/- 16 minutes, intensive care unit stay was 22 +/- 7 hours, and hospital stay was 6.4 +/- 1.2 days. Median postoperative blood loss was 332 +/- 104 mL. There was 1 hospital death. On follow-up at 16.4 +/- 12.2 months, there was no late death or reoperation. New York Heart Association functional class improved from 2.6 +/- 0.5 to 1.4 +/- 0.8. Use of video and robotic assistance minimized incision length and allowed visualization of the whole mitral valve apparatus. The transthoracic clamp facilitated aortic crossclamping and injection of cardioplegia. These findings indicate that the procedure is safe and effective and suggest advantages over conventional surgery in terms of cost, cosmesis, blood loss, postoperative discomfort, intensive care unit and hospital stay. PMID- 12079937 TI - Total reconstruction of the mitral valve with autopericardium: anatomical study. AB - Mitral valve repair has several advantages over prosthetic valve replacement. A new technique of total reconstruction of the mitral valve with autologous pericardium is described. The native mitral valve leaflets and chordae were excised from 10 human cadaver hearts, in the same way as for prosthetic valve replacement. The dimensions of the physiologically normal mitral valve were used to calculate the parameters for tailoring a corresponding new valve. Autologous pericardium was fixed in 0.625% glutaraldehyde solution for 10 minutes. The calculated parameters of the mitral valve were marked on the pericardium. The new valve was fashioned and inserted in the native valve position. Hydraulic probes showed good competence in all 10 reconstructed mitral valves. This method might be a good alternative to prosthetic valve replacement. PMID- 12079938 TI - Thoracofemoral bypass for treatment of juxtarenal aortic occlusion. AB - Descending thoracic aorta-to-femoral artery bypass grafting is considered a good alternative procedure for revascularization in cases of aortic graft failure, graft infection, and other intraabdominal pathologies not amenable to standard aortofemoral revascularization. Its use as the primary mode of treatment in selected cases is still under investigation. From January 1998 to June 2001, 5 patients underwent descending thoracic aorta-to-femoral artery bypass grafting as primary treatment for juxtarenal aortic occlusion. There was no operative mortality nor major morbidity; a groin incision infection occurred in one case. The mean hospital stay was 8.2 days and intensive care unit stay was 2.6 days. Graft failure was not encountered in the short-term follow-up. In spite of the small number of patients, it was concluded that thoracic aortofemoral bypass offers excellent inflow and reliable patency and may be considered for primary revascularization in cases of juxtarenal aortic occlusion. PMID- 12079939 TI - Management of sternal fractures: 239 cases. AB - A review of the management of 239 patients with sternal fractures in a busy trauma center between October 1989 and May 2000 was undertaken to determine the incidence, significance, morbidity, and mortality of this injury. There were 140 men and 99 women with a mean age of 50.3 years (range, 15 to 93 years). Sternal fractures accounted for 8% of admissions for thoracic trauma. The causes were motor vehicle collisions in 215 patients (90%) and falls or direct blows in 24 (10%). Only 64 of 204 car accident patients (31%), 28 men and 36 women, were restrained by seat belts. Complications developed in 13 patients (5.4%). Mortality rate was 0.8%. Mean length of stay in the ward was 6.4 days (range, 1 to 32 days). Four patients (1.7%) underwent surgery. The results show that isolated sternal fractures have low associated morbidity and mortality. Admission is justified for the management of pain and treatment of cardiac complications and concomitant injuries. PMID- 12079940 TI - Thymoma associated with pure red-cell aplasia: clinical features and prognosis. AB - As information on the clinical features and prognosis of thymoma complicated by pure red-cell aplasia is limited, follow-up data on thymoma patients who had a thymectomy between 1954 and 1999 were analyzed retrospectively. Six of 166 cases were complicated by pure red-cell aplasia. In 3 of these, the pure red-cell aplasia appeared after surgical intervention. Remission was observed in 2 patients who underwent extended thymectomy. The other 4 patients subsequently died from pure red-cell aplasia. The outcome in patients with pure red-cell aplasia was poorer than that in the entire group of patients with thymoma and in those with thymoma complicated by myasthenia gravis. The possible onset of pure red-cell aplasia after thymectomy should be kept in mind during follow-up. PMID- 12079941 TI - Right heart assist during beating bypass for severe left ventricular dysfunction. AB - Three patients with triple-vessel disease and severe left ventricular dysfunction underwent successful revascularization on a beating heart, using a right ventricular assist system. Heparin-coated circuits with a centrifugal pump provided 2.5 to 3.5 L x min(-1) of flow to maintain good hemodynamics and enable easy access to the posterior vessels during vertical displacement of the heart. PMID- 12079943 TI - Coronary bypass surgery in patient with malaria. AB - A 65-year-old man with unstable angina pectoris developed malaria prior to coronary artery bypass grafting. After 3 weeks on antimalarial therapy, left internal mammary artery-to-left anterior descending artery anastomosis was performed on the beating heart to avoid the effects of cardiopulmonary bypass. There was no complication in the early postoperative period. PMID- 12079944 TI - "Y" graft bypass for bilateral coronary ostial aortoarteritis. AB - A case of bilateral coronary ostial aortoarteritis, which presented with angina pectoris, is reported. Emergency total arterial revascularization was performed using the bilateral mammary artery and radial artery, and the radial artery was hanged "Y" on the left internal mammary artery. The patient was discharged on low dose steroid. He was asymptomatic at 1-year follow-up. PMID- 12079945 TI - Acute thrombotic obstruction of mitral valve prosthesis: low protein C level. AB - A 51-year-old female underwent redo mitral valve replacement with a pericardial bioprosthesis because of acute thrombotic obstruction of a mechanical valve, in spite of adequate anticoagulation with warfarin. Her protein C level was 24% of the normal value and protein S was reduced to 54% of normal. PMID- 12079946 TI - Isolated idiopathic pulmonary artery aneurysm. AB - Aneurysm formation of the main pulmonary artery is rare. Its natural history is not well understood and there are no clear guidelines on optimal treatment. A 20 year-old woman with a huge saccular aneurysm of the main pulmonary artery, underwent repair with a pericardial patch and concomitant reconstruction of the pulmonary valve. The patient was doing well on follow-up at 6 months; echocardiography revealed a good repair with mild to moderate pulmonary regurgitation. PMID- 12079948 TI - Carotid body tumors. AB - Carotid body paragangliomas were diagnosed by Doppler ultrasound, carotid artery angiography, and cranial computed tomography in a 35-year-old man with a mass in the neck and hearing loss, and in a 42-year-old man with headache, syncope, and a mass in the neck. They underwent successful surgical excision. PMID- 12079947 TI - Intramural cardiac myxoma in left ventricular wall: an unusual location. AB - A 27-year-old asymptomatic woman was investigated for an abnormal finding in the cardiac contour on routine chest radiography. Echocardiography revealed a heterogeneous mass in the anterolateral left ventricular wall. Excision of the tumor disclosed an absence of communication between the residual cavity and the left ventricular endocardium. Histological features of the tumor indicated an intramural myxoma. PMID- 12079949 TI - Giant aneurysm of renal artery: surgical management. AB - Giant aneurysm of the renal artery is rare even though renal artery aneurysms are diagnosed more often since the introduction of abdominal ultrasonography and selective renal arteriography. A 52-year-old man with an aneurysm of the left renal artery measuring 16 x 13 x 10 cm presented with features of an expanding aneurysm. He underwent resection of the aneurysm and a left nephrectomy. PMID- 12079950 TI - Gas gangrene in patient with atherosclerosis obliterans. AB - Clostridia are the main cause of nontraumatic spontaneous gas gangrene. Poor blood flow due to arterial occlusion exacerbates the anaerobic condition. Fulminant gas gangrene in a 54-year-old man with atherosclerosis obliterans was treated by revascularization of the iliac artery using endarterectomy, and his gangrenous lower leg was amputated. However, he died from renal failure. PMID- 12079951 TI - St. Jude medical valve in pulmonary position: anticoagulation and thrombosis. AB - Between 1979 and 1985, 5 patients underwent pulmonary valve replacement with a St. Jude Medical valve. During follow-up ranging from 30 months to 18 years, there were 6 episodes of valve thrombosis in 4 patients. Three episodes were treated by thrombolysis. Reoperation was necessary in 3 patients. Although the St. Jude Medical valve in the pulmonary position can occasionally function long term without anticoagulation, these cases show the high risk of thrombosis and need for anticoagulation. PMID- 12079952 TI - Successful repair of ascending aortic aneurysm due to Takayasu's arteritis. AB - A 2-year-old girl with Takayasu's arteritis and an ascending aortic aneurysm underwent successful graft replacement. Although aneurysms of the great vessels are rarely encountered, this disease should be considered in the differential diagnosis of ascending aortic aneurysms in children. PMID- 12079953 TI - Primary lymphoepithelioma-like carcinoma of lung. AB - A 57-year-old Caucasian woman presented with nonproductive cough. Computed tomography revealed a peripheral solid mass in the upper lobe of the left lung. She underwent thoracotomy and upper lobectomy. Histology of the excised tumor demonstrated lymphoepithelioma-like carcinoma of the lung, with no associated Epstein-Barr virus activity. Being a rare entity and mostly seen in Asians, very few cases have been described previously. PMID- 12079954 TI - Mediastinal myelolipoma. AB - Myelolipoma is a rare tumor, and a mediastinal location is extremely unusual. The main pathologic feature is the coexistence of mature adipose tissue and bone marrow cells; the presence of megakaryocytes is essential for diagnosis. The successful removal of a mediastinal myelolipoma in a 59-year-old man is described. PMID- 12079955 TI - Bronchial artery dissection and fatal hemothorax following pneumonectomy. AB - A 59-year-old man died suddenly and unexpectedly two days after an uncomplicated pneumonectomy for bronchogenic adenocarcinoma. In addition to a distal pulmonary thromboembolism found at postmortem, there was significant hemorrhaging into the pneumonectomy space, which was associated with rupture of a dissecting aneurysm in the calcified bronchial artery stump. The pathology and surgical implications of bronchial artery disease are discussed. PMID- 12079956 TI - A technique facilitating muscle-sparing thoracotomy. AB - The muscle-sparing thoracotomy technique preserves the latissimus dorsi and serratus anterior muscles and provides excellent exposure for most thoracic and mediastinal operations. It also reduces postoperative pain and complications, and preserves pulmonary function. The technique has been developed further to facilitate subcutaneous dissection by insufflation of air from a syringe connected to a large-bore needle via a 3-way tap. PMID- 12079957 TI - Multiple mycotic aortic aneurysms in a drug addict. PMID- 12079958 TI - Type B aortic dissection and right hemothorax. PMID- 12079959 TI - Left atrial reduction: isolation of the pulmonary veins. PMID- 12079960 TI - The Rosetta Stone and cardiac surgery. PMID- 12079961 TI - Cardiac surgery under perfusionless hypothermia: Siberian experience. AB - Between January 1995 and December 1999, 942 patients (452 males and 490 females) aged 1 to 51 years underwent definitive surgery under perfusionless hypothermia for correction of congenital heart defects, predominantly uncomplicated ventricular or atrial septal defects (80%). Hypothermia of 24 degrees C to 28 degrees C was achieved in 15 to 45 minutes (mean, 25.7 +/- 1.2 minutes) by application of crushed ice over the body and head. Aortic crossclamp time ranged from 10 to 76 minutes (mean, 26.1 +/- 0.25 minutes). Cardiac restoration time ranged from 1 to 10 minutes (mean, 2.1 +/- 0.08 minutes). Eight patients (0.85%) died postoperatively: 4 from acute cardiac insufficiency, 2 as a consequence of technical faults, 1 from persistent pulmonary hypertension, and 1 had sudden cardiac arrest. None of the surviving patients showed any gross neurological deficit. Perfusionless hypothermic cardiac surgery, when applied appropriately, is safe and simple, and might still have a place in treating a selected group of patients with uncomplicated congenital heart defects. PMID- 12079962 TI - Cardiac myxoma and myxosarcoma: clinical experience and immunohistochemistry. AB - From January 1978 to February 1999, 120 patients (42 males and 78 females) with cardiac myxoma (115) or myxosarcoma (5) underwent surgical excision or biopsy. There were 5 early postoperative deaths (mortality, 4.2%). Seventy-three survivors were followed up for 0.75 to 20.25 years (mean, 9.42 years); they comprised 4 myxosarcoma patients who all had recurrence or metastasis, and 69 myxoma patients who had no evidence of recurrence or metastasis. Neither familial myxoma nor Carney complex was found. The 5 cases of myxosarcoma and 18 randomly selected cases of myxoma were evaluated for proliferative activity, metastatic potential, and oncogene products by immunohistochemistry. The expression of p53 and Bcl-2 was similar in both groups. Overexpression of proliferating cell nuclear antigen and low expression of nm23 in myxosarcoma are consistent with the high rate of recurrence and metastasis of this tumor. Surgical resection of sporadic myxoma is a safe and effective treatment with satisfactory early and long-term results. However, the prognosis of myxosarcoma is still disappointing. Regular echocardiography and chest radiography or computed tomography are necessary for early detection of recurrence or metastasis of myxosarcoma. PMID- 12079963 TI - Outcome of cardiac and thoracic aortic operation in patients over 80 years old. AB - A retrospective analysis was performed to determine the early outcome of cardiac and thoracic aortic surgery in patients over 80 years old. Between 1994 and 2000, 41 octogenarians (mean age, 82.6 +/- 2.5 years) underwent coronary artery bypass grafting (25), valve surgery (8), thoracic aortic aneurysm repair (7), or combined valve and bypass surgery (1). Overall hospital mortality was 9.8%. Mortality rates for specific procedures were 12% for coronary bypass, 0% for valve surgery, and 14% for thoracic aortic aneurysm repair. Major postoperative complications affected 27% of patients and included severe low cardiac output, respiratory failure, and acute renal failure, with a low incidence of perioperative stroke (2.4%). Cardiac and thoracic aortic operations can be performed with acceptable mortality and morbidity when appropriately applied in selected octogenarians. PMID- 12079964 TI - Transfusion for coagulopathy after heart surgery: efficacy of laboratory studies. AB - Limited information exists regarding transfusions specifically for coagulopathy following cardiac surgery and the value of pre- and postoperative coagulation tests. Procedures (86% coronary bypass, 7.5% valve, and 6.5% combined valve and bypass) on 478 patients were reviewed; 101 patients (21%) were transfused for postoperative coagulopathy. Compared to those not transfused, patients with coagulopathy were significantly older and smaller, and they had more combined valve procedures, emergency operations, and preoperative heparin treatment as well as longer crossclamp and bypass times. Three preoperative tests showed significant differences in the coagulopathy group: activated clotting time, partial thromboplastin time, and antithrombin-III level. Four postoperative tests showed significant differences between the groups: prothrombin time, partial thromboplastin time, fibrinogen level, and fibrin split products at 10 dilutions. Patient characteristics and pre- and postoperative testing can identify patients at high risk of transfusion specifically related to coagulopathy. PMID- 12079965 TI - Routine use of the radial artery for coronary artery revascularization. AB - Between January 1997 and December 2000, a total of 4,000 patients underwent myocardial revascularization using the radial artery as one of the conduits. The mean age of the patients was 54 +/- 7 years, and 92.8% of them were male. Of these patients, 31% had a left ventricular ejection fraction below 40% and 22.8% underwent urgent operation. A total of 4,225 distal anastomoses were performed using the radial artery. The average number of grafts was 3.3 +/- 0.5. The hospital mortality rate was 0.8%. Low cardiac output, inotropic support, perioperative myocardial infarction, reoperation for bleeding, atrial fibrillation, and sternal infection occurred in 1.8%, 2.8%, 1.2%, 1.2%, 16.8%, and 1.2% of the patients, respectively. None of the patients had major ischemia of the hand. The incidence of local hand wound complications was 0.7% (wound infection, 0.4%; wound dehiscence without infection, 0.1%; and hematoma, 0.2%). The average length of stay in the intensive care unit was 20 +/- 7 hours and in the hospital was 6 +/- 2 days. Postoperative angiography, performed in 106 patients at a mean interval of 18 months, showed that 92.4% of radial artery, 96.2% of internal mammary artery, and 76.2% of saphenous vein grafts were patent. PMID- 12079966 TI - Tissue engineering of an auto-xenograft pulmonary heart valve. AB - To improve the durability of stentless valves without losing their excellent hemodynamic function, a new-generation auto-xenograft was developed and evaluated. A piece of vein was harvested from 3 juvenile sheep 6 weeks before implantation of the valve. Endothelial cells from the vein material were cultivated and used to reendothelialize a decellularized porcine pulmonary valve. The tissue-engineered valve was implanted into the right ventricular outflow tract of the juvenile sheep. It was explanted after 100 days and assessed macroscopically as well as by x-ray, light microscopy (hematoxylin and eosin staining and von Kossa staining), and scanning electron microscopy. Calcium content of the cusps was determined quantitatively by atomic absorption spectrometry. The sheep implanted with the valve recovered quickly without any problems during the observation period. X-ray examination of the 3 explanted valves showed no cusp calcification, which was confirmed by histological study. Atomic absorption spectrometry showed low tissue calcium content. A clinical safety and feasibility trial with an allograft valve prepared the same way showed excellent short-term results in 6 patients. PMID- 12079967 TI - Play safe: band the late presenting complete atrioventricular canal. AB - Primary repair of complete atrioventricular canal in patients who present beyond one year of age carries a high mortality. Between January 1995 and February 2000, 16 patients aged 8 to 24 months (mean, 14.5 months) received pulmonary artery banding at presentation and underwent total correction at 24 to 96 months old (mean, 41.9 months). There was one hospital death (mortality, 6.25%). During a mean follow-up of 10.2 months (range, 6 to 28 months), there was no late death, 13 of the 15 survivors (87%) were in New York Heart Association functional class I, and 2 (13%) were in class III. In patients with complete atrioventricular canal who present late with severe reactive pulmonary hypertension, banding followed by complete repair reduces the risk associated with primary repair. PMID- 12079969 TI - Early and long-term results of surgical repair of pectus excavatum. AB - From 1990 to 1998, 30 patients underwent surgery for correction of pectus excavatum. There were 19 (63%) males and 11 (37%) females, aged 4 to 32 years (mean, 12.57 years). Bilateral excision of 4 to 6 costal cartilages and sternal wedge osteotomy were performed on 27 (90%) patients, and Kirschner wires were used for substernal support in 25 (83%). A median sternotomy was carried out in males and a submammary transverse incision was preferred in females. All patients were followed up at yearly intervals. Early results were excellent in all except 2 cases: a 14-year-old boy developed contralateral sternal depression after costochondral excision without sternal elevation for correction of one-sided costochondral hyperplasia; and an 8-year-old girl in whom no Kirschner wires had been inserted developed recurrent minimal sternal depression. Long-term follow-up showed recurrent sternal depression 6 years postoperatively in a boy who had undergone surgery at 4 years old, with early removal of the Kirschner wires. It is recommended that correction of pectus excavatum should be carried out in prepubertal children, and Kirschner wires should be used for substernal support. PMID- 12079968 TI - Total cavopulmonary connection with extraatrial tunnel. AB - Between April 1997 and February 2000, total cavopulmonary connection with an extraatrial tunnel was used to treat 9 cases of complicated congenital heart disease: single ventricle (4), double-outlet right ventricle (3), mitral atresia (1), and tricuspid atresia (1). There was no mortality. One patient developed bacterial endocarditis and required reoperation after 52 days to replace the tunnel. At follow-up ranging from 11 months to 3 years, 3 patients were in New York Heart Association functional class I, and 6 were in class II. One patient with single ventricle had refractory supraventricular tachycardia after a modified Fontan operation 4 years earlier, which was cured by the total cavopulmonary connection procedure. The essential factors for a good outcome include appropriate surgical indication, avoidance of aortic crossclamping and cardiac arrest, and unobstructed anastomosis between the superior and inferior venae cavae and the pulmonary artery. PMID- 12079970 TI - Common causes of pleural effusion in referral hospital in Isfahan, Iran 1997 1998. AB - During a one-year period to September 1998, data were collected from all 213 patients referred with pleural effusion. There were 132 males and 81 females; their ages ranged from 18 to 85 years. The most common etiologies of effusion were congestive heart failure (39.4%), malignancy (27.2%), pneumonia (8%), empyema (5.2%), and tuberculosis (5.2%). Pleural effusions are frequent in Iran, and the causes are fairly similar to those reported by European authors, but with slightly more tuberculosis cases, mostly among Afghan refugees. PMID- 12079971 TI - Transcatheter coil embolization of coronary artery fistula. AB - Between December 1982 and August 2001, coil embolization of coronary artery-to cardiac chamber fistula was attempted in 15 patients aged 11 months to 44 years (mean, 7.2 +/- 2.5 years). The fistulae connected the left anterior descending artery to the right ventricle in 4 patients, the right coronary artery to the right ventricle in 3, the right coronary artery to the right atrium in 3, the circumflex artery to the right ventricle in 2, the circumflex artery to the right atrium in 2, and the right coronary artery to the trunk of the pulmonary artery in 1. Complete fistula occlusion was achieved in 14 patients (93%); one had a residual shunt and underwent repeat embolization one year later, resulting in complete occlusion. There was one early death (7%) in a 4-year-old girl who developed femoral artery thrombosis and acute renal failure. Complications comprised migration of the coil into the pulmonary artery (2), femoral artery thrombosis (2), and perforation of the vessel wall by the guidewire (1) with immediate thrombosis and occlusion of the fistula (no coil was deployed). The 13 survivors with coils were followed up for 0.5 to 13 years; complete occlusion of the fistula was confirmed in all cases. PMID- 12079972 TI - Situs inversus and coronary artery disease. AB - Situs inversus is a rare condition and there are few reports of myocardial revascularization in such patients. A 56-year-old woman with situs inversus totalis and coronary artery disease underwent successful anastomosis of the right internal mammary artery to the anterior descending coronary artery, and a saphenous vein graft to the right coronary artery. PMID- 12079973 TI - Multiple giant aneurysms of coronary arteries. AB - Multiple giant aneurysms of the coronary arteries occur extremely rarely and are usually associated with Kawasaki disease in children. A 53-year-old man with multiple giant coronary artery aneurysms was treated successfully by aortocoronary bypass grafting. PMID- 12079974 TI - Emergency operation for spontaneous coronary artery dissection in young female. AB - Emergency coronary artery bypass grafting using a composite graft of the right internal thoracic artery and the radial artery was successfully performed for impending myocardial infarction secondary to spontaneous dissection of the right coronary artery in a 46-year-old woman. As spontaneous coronary artery dissection frequently presents as myocardial infarction or sudden death, timely diagnosis and intervention are required. PMID- 12079975 TI - Superior mesenteric artery aneurysm. AB - A 38-year old woman with mid-epigastric pain, diarrhea, and weight loss, underwent resection of a superior mesenteric artery aneurysm and primary repair of the artery. Pathological examination showed degenerative atherosclerotic changes, marked medial and intimal thickening, and vegetations. Microbiological studies demonstrated Streptococcus viridans as the infecting organism of this mycotic aneurysm. The patient made a good recovery and remained well after 3 years. PMID- 12079976 TI - Lipoma of the left ventricle. AB - A 36-year-old female was admitted with dyspnea on exertion of one year's duration. Echocardiography revealed a tumor arising from interventricular septum with dynamic left ventricular outflow tract obstruction. On cardiopulmonary bypass with cardioplegic arrest, the tumor was approached through a transverse aortotomy, and excised from the interventricular septum through the aortic valve. Postoperative recovery was uneventful and the patient was asymptomatic with no recurrence at follow-up after one year. PMID- 12079977 TI - Surgical treatment of echinococcal cysts of the heart: report of 3 cases. AB - Three patients diagnosed with hydatid cysts of the heart underwent surgical treatment. The cysts were enucleated and the cavities were closed in 2 cases by plication and obliteration. In one case, the cystic cavity was closed with biologic glue. The outcome was satisfactory in all 3 patients. Postoperative treatment with albendazole is recommended. PMID- 12079978 TI - Percutaneous extraction of entrapped infective transvenous pacing lead. AB - A 57-year-old woman had an entrapped infected transvenous pacing lead successfully removed percutaneously with a Dormier basket under fluoroscopy, thus avoiding a major surgical procedure. PMID- 12079979 TI - Iatrogenic coarctation after coil occlusion of arterial duct. AB - Coil occlusion of a patent ductus arteriosus was performed in an 8.5-month-old girl with a large left-to-right shunt through a wide arterial duct. Post occlusion echocardiography revealed iatrogenic obstruction of the aorta caused by protrusion of the loops of the Jackson coil into the descending aorta. The problem resolved spontaneously during follow-up. PMID- 12079981 TI - Congenital pulmonary lymphangiectasia with chylothorax. AB - A 5-year-old boy with dyspnea was diagnosed through thoracentesis as having chylothorax. After conservative management failed, open thoracotomy was performed. Lung biopsy confirmed the diagnosis of congenital pulmonary lymphangiectasia. Chylothorax was successfully controlled by fibrin glue pleurodesis in this patient. PMID- 12079982 TI - Unexplained systemic hypertension after closure of ductus arteriosus. AB - Immediately after surgical closure of a patent ductus arteriosus, a 12-year-old boy developed severe systemic arterial hypertension refractory to medication. The cause of hypertension could not be found, but it came under control 3 weeks postoperatively with a combination of angiotensin-converting enzyme inhibitor and chlorothiazide. PMID- 12079983 TI - Right pulmonary artery-to-left atrial fistula. AB - Right pulmonary artery-to-left atrial fistula is a rare congenital anomaly. Contrast echocardiography and cardiac catheterization diagnosed a right pulmonary artery-to-left atrial fistula in a 24-year-old female with a history of cyanosis since childhood, and recurrent thromboembolism. She underwent successful surgical closure of the fistula. PMID- 12079984 TI - Cardiopulmonary bypass before general anesthesia in prosthetic valve thrombosis. AB - Valve obstruction is a lifethreatening complication of mechanical valve prostheses. Emergency operation is mandatory for patients who subsequently develop cardiogenic shock and severe pulmonary edema. In this severely compromised hemodynamic condition, cardiac arrest develops in most of the patients before surgery and just after general anesthesia induction. In one such case, we performed femorofemoral cardiopulmonary bypass with local anesthesia before general anesthesia induction and successfully replaced the thrombosed prosthetic valve, thus avoiding a catastrophic outcome. PMID- 12079985 TI - Spinal cord ischemia associated with cardiac arrest. AB - Ischemic spinal cord damage from cardiac arrest is rare. A 48-year-old man underwent a straightforward retroperitoneal aortofemoral bypass operation. The following day, he developed cardiac arrest and was resuscitated successfully, but permanent paraplegia was observed soon afterwards. PMID- 12079986 TI - Esophageal perforation caused by a blister-wrapped tablet. AB - An 84-year-old man was diagnosed with esophageal perforation following ingestion of a blister-wrapped tablet. His condition improved after 2 weeks of conservative treatment using antibiotics and high-protein enteral nutrition. PMID- 12079987 TI - Power Doppler imaging for detection of harvest injury of internal mammary artery. AB - Harvest injury of the internal mammary artery can lead to early thrombosis, which may be difficult to differentiate from atherosclerosis or harvest spasm in some cases. We describe the efficacy of intraoperative evaluation of the internal mammary artery by power Doppler imaging during off-pump or minimally invasive direct coronary artery bypass grafting. PMID- 12079988 TI - Totally robotic technique in multivessel coronary disease--is it possible? AB - Robotic techniques in coronary surgery have been established for a few years. The preparation of the internal mammary arteries is now a well-described procedure, as is anastomosis to the anterior wall vessels under cardiopulmonary bypass. To achieve totally robotic complete revascularization of the heart, there are still some problems to be solved. Potential solutions are discussed. Current techniques using telemanipulation systems in cardiac surgery are described and explained. Totally endoscopic off-pump complete revascularization of the heart might be achieved by a combination of techniques already developed by different teams around the world. PMID- 12079989 TI - Closure of bronchial defects using glutaraldehyde-treated pericardium. PMID- 12079991 TI - Bidirectional Glenn shunt: 170 cases. PMID- 12079990 TI - Delayed sternal closure after cardiac operations. PMID- 12079992 TI - Molecular cloning and characterization of oppo 1: a haploid germ cell-specific complementary DNA encoding sperm tail protein. AB - We isolated a cDNA clone specifically expressed during spermatogenesis from a subtracted cDNA library of mouse testis. The cDNA consisted of 1085 nucleotides and had an open reading frame of 870 nucleotides encoding a putative protein of 290 amino acid residues. Northern blot analysis revealed a 1.2-kilobase mRNA exclusively expressed in the testis in adult mice; the mRNA was first detected late pachytene stage, and expression increased as the animals matured. The protein encoded by the mRNA had a molecular weight of approximately 33 kDa by Western blot analysis, and was localized to occupy the flagella from the connecting piece through the principal piece. We named this newly isolated gene oppo 1, and we suggest that it plays an important role in sperm tail structure and/or sperm movement. PMID- 12079993 TI - Luteotropic roles of prolactin in early pregnant hamsters. AB - Prolactin (PRL) has long been regarded as a luteotropin maintaining early pregnancy in rodents. To delineate luteotropic roles of PRL in terms of luteal vascularization and immune privilege, luteal expression of Thy-1 differentiation protein, Fas, and Fas ligand (FasL) in early pregnancy was studied in hamsters on Day 4 of pregnancy (P4 group). Release of pituitary PRL was blocked by daily treatment with bromocriptine (1 mg s.c. given at 1000 h) on Days 1-4 of pregnancy (PB group). PRL withdrawal induced functional luteolysis, as evidenced by a precipitous drop in serum progesterone to background levels. In situ 3' end labeling of fragmented DNA (TUNEL method) also clearly showed that many apoptotic nuclei accumulated in the disintegrated luteal vessels in the corpus luteum in the PB group. Immunohistochemical studies showed that luteal Thy-1-positive vascular pericytes were abundant in the P4 group but rare in the PB group. Thus, PRL is essential for luteal vascularization in early pregnancy. Western blotting and quantitative real-time reverse transcription-polymerase chain reaction data showed that Fas protein and mRNA levels increased, whereas those of FasL decreased after PRL withdrawal. Accordingly, apoptosis initiated by Fas-FasL interaction is involved in the bromocriptine-induced luteolysis. Therefore, luteotropic roles of PRL are to support Thy-1 positive pericytes in maintaining proper luteal vascularization and to prevent immune insult by preserving a balance between luteal Fas and FasL expression in early pregnancy. PMID- 12079994 TI - Activin A, estradiol, and free insulin-like growth factor I in follicular fluid preceding the experimental assumption of follicle dominance in cattle. AB - In cattle, the two largest follicles of a wave (F1, F2) begin to deviate into a dominant follicle and a subordinate follicle when F1 is a mean of 8.5 mm in diameter. After the beginning of deviation, F1 and F2 are diameter-defined dominant and subordinate follicles. Changes associated with the conversion of F2 into a future dominant follicle were studied by ablating F1 at the expected beginning of deviation (F1, 8.5 mm; Hour 0) and assessing the follicular-fluid factors in F2. Follicles were designated F1C and F2C in controls and F2A in F1 ablated heifers. Follicular-fluid collections were made at Hours 0, 4, 8, or 12 (n = 7 heifers per hour; fluid from F1C, F2C, and F2A; experiment 1) or at Hours 4, 6, 8, 10, or 12 (n = 9 heifers per hour; fluid from F2A; experiment 2). Postablation concentrations of circulating FSH increased (P < 0.05) between Hours 2 and 6. Diameter of F2A increased (P < 0.05) after Hour 8 in both experiments so that the diameter of F2A at Hours 10 or 12 was not different (P > 0.1) from the diameter of F1 at Hour 0. A transient elevation (P < 0.05) in follicular-fluid activin A occurred in F2A at Hour 8 in both experiments. Concentrations of estradiol (P < 0.05) and insulin-like growth factor I (IGF-I; P < 0.1) decreased in F2C by Hour 8. In F2A, the concentrations of both factors began to increase (P < 0.05) after Hours 4 or 8 so that there was no difference (P > 0.1) between F1C and F2A at Hour 12. Concentrations of IGF-I and IGF binding protein 2 (IGFBP-2) in F2A changed in opposite directions at the same hours. No differences between follicles were found for concentrations of progesterone, androstenedione, inhibin A, and inhibin B. The order of events in the conversion of a future subordinate follicle to a future dominant follicle was an increase in systemic FSH, a transient elevation in follicular-fluid activin A, and a simultaneous increase in follicular-fluid estradiol and restoration of an apparent growth-compatible balance of free IGF-I and IGFBP-2. PMID- 12079996 TI - Positive regulation of retinoic acid receptor alpha by protein kinase C and mitogen-activated protein kinase in sertoli cells. AB - Retinoic acid receptor alpha (RARalpha) is required for normal testis function. Similar to other steroid hormone receptors, RARalpha appears to undergo an activation process by which it translocates from the cytoplasm to the nucleus where it acts as a transcription factor. In this report, we demonstrate that RARalpha nuclear trafficking in Sertoli cells is positively regulated by phorbol 12-myristate-13-acetate-activated protein kinase C without the requirement of ligand, retinoic acid. Protein kinase C then stimulates the downstream mitogen activated protein kinase, and the nuclear localization of RARalpha is dependent on activation of both kinases. The increase in RARalpha nuclear translocation is also coupled with enhanced transcriptional activity of RARalpha. This mechanism of RARalpha positive regulation is unique, different from that of its negative regulation, that has previously been shown to be dependent on cAMP-dependent protein kinase A and more importantly, dependent on its ligand. However, the mechanism by which retinoic acid positively influences the nuclear localization of RARalpha is not due to retinoic acid directly increasing protein kinase C or mitogen-activated protein kinase activities. Nonetheless, the positive influence of retinoic acid is also dependent on these two kinases as determined by inhibitor studies. These results suggest two mechanisms for RARalpha activation in Sertoli cells: one involving only the two kinases, the other involving both the ligand and the two kinases. These regulatory mechanisms for RARalpha activation, both positive and negative, may be critical for the proper function of RARalpha in the testis. PMID- 12079995 TI - Gene therapy of erectile dysfunction in the rat with penile neuronal nitric oxide synthase. AB - Gene transfer to the penile corpora cavernosa of constructs of the inducible and endothelial nitric oxide synthase (NOS) cDNAs ameliorates erectile dysfunction in aged rats. In this study, we investigated whether the neuronal NOS (nNOS) variant responsible for erection, penile nNOS (PnNOS), can exert a similar effect, and whether the combination of electroporation with a helper-dependent adenovirus (AdV) improves gene transfer. PnNOS and beta-galactosidase cDNAs were cloned in plasmid (pCMV-PnNOS; pCMV-beta-gal) and "gutless" AdV (AdV-CMV-PnNOS; AdV-CMV beta-gal) vectors, and injected into the penis of adult (beta-gal) or aged (PnNOS) rats, with or without electroporation. Penile erection was measured at different times after PnNOS cDNA injection, by electrical field stimulation of the cavernosal nerve. The expression of beta-galactosidase or PnNOS was estimated in penile tissue by either histochemistry and luminometry or Western blot, and the effects of AdV-CMV-PnNOS on mRNA expression were examined by a DNA microarray. We found that electroporation increased pCMV-beta-gal uptake, and its expression was detectable at 56 days. In the aged rats treated with pCMV-PnNOS and electroporation, the maximal intracavernosal:mean arterial pressure ratios were elevated for 11 and 18 days when compared with those in controls. Electroporation intensified penile uptake of as few as 10(6) viral particles (vp) of AdV-CMV-beta-gal, and with 10(7) vp beta-galactosidase was still detectable at 60 days. Electroporated AdV-CMV-PnNOS (10(7) vp) was effective at 18 days in stimulating the erection of aged rats, without inducing the expression of cytotoxic genes. In conclusion, intracavernosal gene therapy with PnNOS cDNA corrected the aging-related erectile dysfunction for at least 18 days when given by electroporation in a helper-dependent AdV at low viral loads. PMID- 12079997 TI - Alterations in peripheral concentrations of inhibin A in cattle studied using a time-resolved immunofluorometric assay: relationship with estradiol and follicle stimulating hormone in various reproductive conditions. AB - The aims of this study were to develop a sensitive and specific assay for bovine inhibin A using europium and to investigate the endocrine role of inhibin A in various reproductive conditions by characterizing the relationship between profiles of inhibin A, FSH, and estradiol and follicle growth during the postpartum period, during the intact estrous cycle, and in cows with follicular cysts. The time-resolved immunofluorometric assay (Tr-IFMA) for bovine inhibin A, using purified polyclonal antibodies to alpha and beta(A) subunits, was specific for bovine inhibin A and did not cross-react with bovine activin A, activin AB, activin B, pro-alphaC or human recombinant inhibin B. The detection limit of the IFMA was 3.3 pg/ml expressed in terms of bovine 32-kDa inhibin A. Dose-response curves of plasma samples obtained from intact and FSH-stimulated cows and cystic cows were parallel to the standard without any preassay processing of samples. Plasma inhibin A levels increased (P < 0.01) concomitant with emergence of nonovulatory or ovulatory follicular waves during the postpartum period. In cystic cows, plasma inhibin A was sustained at high levels for a longer period, associated with growth of persistent dominant follicles. The highest levels of inhibin A were noted during the growth phase of normal and persistent dominant follicles; however, inhibin A levels declined (P < 0.01) as these dominant follicles ceased to grow or ovulated. An inverse relationship between patterns of plasma inhibin A and FSH existed during each follicular wave in the three physiologic conditions. Increases in plasma inhibin A levels were associated with increases in plasma estradiol levels during most follicular waves; however, there was no increase in plasma estradiol level and no relationship between patterns of estradiol and FSH during follicular waves observed during the early postpartum period or midluteal phase of the estrous cycle. In conclusion, the Tr-IFMA does not require pretreatment of samples and can be used for precise measurement of bovine inhibin A without interference with free inhibin alpha subunits. Inhibin A, produced primarily during growth of the dominant follicle, functions as a negative feedback regulator for FSH secretion throughout the postpartum period and the estrous cycle, whereas estradiol appears to have a minor role in regulation of FSH compared with inhibin A, especially during the early postpartum period and midluteal phase of the estrous cycle. The results also indicate that a persistent dominant follicle sustains inhibin A production for a longer period than the dominant follicle emerging in the estrous cycle and establishes long term dominance by suppressing emergence of a new follicular wave. PMID- 12079998 TI - A BK(Ca) to K(v) switch during spermatogenesis in the rat seminiferous tubules. AB - Spermatogenesis is a complex cellular event during which the diploid germ cells differentiate and divide by mitosis and meiosis at specific time points along the spermatogenic cycle to generate the haploid spermatozoa. For this complex event to go in an orderly manner, cell differentiation and division must be precisely controlled by signals arising from within and outside the seminiferous tubules. Changes in the membrane potential of the germ cells are likely to be an important part of the signaling mechanism. We have applied the whole-cell patch clamp technique to identify and characterize ion channels in different spermatogenic cells from immature and mature rat testes fractionated by discontinuous Percoll gradient. A voltage- and Ca(2+)- dependent, outwardly rectifying current with gating and pharmacologic properties resembling the large conductance K(+) channels (BK(Ca)) was recorded from the spermatogonia and primary spermatocytes. Another voltage-dependent, outwardly rectifying current that was sensitive to 4 aminopyridine, a K(v) channel blocker, was detected in spermatocytes and early spermatids. This current is likely caused by the smaller conductance, voltage sensitive K(+) channels (K(v)). In some spermatogonia, both the BK(Ca) channels and the K(v) channels could be simultaneously detected in the same cell. It appears that during the course of spermatogenesis, there is up-regulation of K(v) but down-regulation of BK(Ca). Reverse transcription-polymerase chain reaction, Western blot analysis, and immunohistochemistry further confirmed the differential expression of the ion channels in different spermatogenic cells. We conclude that these ion channels may play an important role in the control of spermatogenesis. PMID- 12079999 TI - Abnormal expression of trophoblast major histocompatibility complex class I antigens in cloned bovine pregnancies is associated with a pronounced endometrial lymphocytic response. AB - Early embryonic losses are much higher in nuclear transfer (cloned) pregnancies, and this is a major impediment to improving the efficiency of cloned animal production. In cattle, many of these losses occur around the time of placental attachment from the fourth week of gestation. We studied the potential for altered immunologic status of cloned pregnancies to be a contributing factor to these embryonic losses. Expression of major histocompatibility complex class I (MHC-I) by trophoblast cells and distribution of endometrial T-lymphocyte numbers were investigated. Six 5-wk-old cloned pregnancies were generated, and 2 others at 7 and 9 wk were also included, all derived from the same fetal cell line. All 8 cloned placentas displayed trophoblast MHC-I expression. None of the 8 controls (4-7 wk old) showed any MHC-I expression. The percentage of trophoblast cells expressing MHC-I varied in the clones from 17.9% to 56.5%. Numbers of T lymphocytes (CD3(+) lymphocytes) were significantly higher in the endometrium of the majority of cloned pregnancies compared with controls. In the cloned pregnancies, large aggregates of T cells were frequently observed in the endometrium in addition to increased numbers of diffusely spread subepithelial lymphocytes. As trophoblast MHC-I expression is normally suppressed during early gestation, the observed MHC-I expression in the cloned pregnancies is likely to have induced a maternal lymphocytic response that would be detrimental to maintaining viability of the cloned pregnancy. These findings support a role for immunologic rejection in the syndrome of early embryonic loss in cloned bovine pregnancies. PMID- 12080000 TI - Activation of protein kinase C induces mitogen-activated protein kinase dephosphorylation and pronucleus formation in rat oocytes. AB - Mammalian oocytes are arrested at metaphase of the second meiotic division (MII) before fertilization. When oocytes are stimulated by spermatozoa, they exit MII stage and complete meiosis. It has been suggested that an immediate increase in intracellular free calcium concentration and inactivation of maturation promoting factor (MPF) are required for oocyte activation. However, the underlying mechanism is still unclear. In the present study, we investigated the role of protein kinase C (PKC) and mitogen-activated protein (MAP) kinase, and their interplay in rat oocyte activation. We found that MAP kinase became dephosphorylated in correlation with pronucleus formation after fertilization. Protein kinase C activators, phorbol 12-myriatate 13-acetate (PMA) and 1,2 dioctanoyl-rac-glycerol (diC8), triggered dephosphorylation of MAP kinase and pronucleus formation in a dose-dependent and time-dependent manner. Dephosphorylation of MAP kinase was also correlated with pronucleus formation when oocytes were treated with PKC activators. Effects of PKC activators were abolished by the PKC inhibitors, calphostin C and staurosporine, as well as a protein phosphatase blocker, okadaic acid (OA). These results suggest that PKC activation may cause rat oocyte pronucleus formation via MAP kinase dephosphorylation, which is probably mediated by OA-sensitive protein phosphatases. We also provide evidence supporting the involvement of such a process in fertilization. PMID- 12080001 TI - Apoptosis, onset and maintenance of spermatogenesis: evidence for the involvement of Kit in Kit-haplodeficient mice. AB - Kit/stem cell factor (SCF ) has been reported to be involved in survival and proliferation of male differentiating spermatogonial cells. This kinetics study was designed to assess the role of Kit/SCF during spermatogenesis in mice, and the extent of male programmed germ cell death was measured between 8 and 150 days of age. For this purpose, 129/Sv inbred mice in which one Kit allele was inactivated by an nlslacZ sequence insertion (Kit(W-lacZ/+)) were compared with 129/Sv inbred mice with wild-type alleles at the Kit locus. Four different approaches were used: 1) morphometric study to assess spermatogenesis, 2) flow cytometry to study testicular cell ploidy, 3) in situ end labeling to detect apoptosis, and 4) follow-up of reporter gene expression. Spermatogenesis was lower in Kit(W-lacZ/+) heterozygous mice both at the onset of spermatogenesis and during adulthood. Indeed, greater apoptosis occurred at the onset of spermatogenesis. This was followed in the adult by a smaller seminiferous tubule diameter and a lower ratio between type B spermatogonia and type A stem spermatogonia in Kit(W-lacZ/+) mice compared with Kit(+/+) mice. These differences are probably related to the Kit haplodeficiency, which was the only difference between the two genotypes. Germ cell counts and testicular cell ploidy revealed delayed meiosis in Kit(W-lacZ/+) mice. Reporter gene expression confirmed expression of the Kit gene at the spermatogonial stage and also revealed Kit expression during the late pachytene/diplotene transition. These results suggest involvement of Kit/SCF at different stages of spermatogenesis. PMID- 12080002 TI - An environmentally relevant organochlorine mixture impairs sperm function and embryo development in the porcine model. AB - We evaluated the effects of an environmentally relevant mixture of more than 15 organochlorines on the development of pig oocytes and sperm during in vitro fertilization (IVF). Oocytes were cocultured with sperm in IVF medium containing increasing concentrations of an organochlorine mixture, similar to that found in women of highly exposed populations. Exposure to the organochlorine mixture diminished oocyte penetration rates and polyspermy in a linear manner. The mixture did not affect rates of cleavage nor development to multicell embryos. However, rates of development to the blastocyst stage were lower at the highest concentration at which oocyte penetration was observed. The same experiment was performed using oocytes that were preexposed during in vitro maturation. This greater exposure to the mixture also reduced penetration in a dose-response manner and affected polyspermy. Frozen-thawed pig sperm were also cultured in IVF medium containing the same organochlorine concentrations. Sperm motility parameters were immediately reduced in a dose-dependent manner by the organochlorines, followed by diminished viability 2 h later. From these results, it appears that reduced sperm quality would account for decreases in fertilization, polyspermy, and blastocyst formation. These results suggest that exposing porcine oocytes and sperm to an environmentally pertinent organochlorine mixture in vitro disrupts the oocyte block to polyspermy, sperm fertility, and further embryonic development, and supports recent concerns that such pollutants harm reproductive health in humans and other species. PMID- 12080003 TI - Potassium channel antagonists influence porcine granulosa cell proliferation, differentiation, and apoptosis. AB - This investigation determined the effects of K(+) channel antagonists on proliferation, differentiation, and apoptosis of porcine granulosa cells. The drugs screened for functional effects included the class III antiarrhythmic agents MK-499 and clofilium, the chromanol I(Ks) antagonist 293B, the benzodiazepine I(Ks) antagonists L-735,821 and L-768,673, and the peptidyl toxins charybdotoxin (CTX) and margatoxin (MTX). Granulosa cell proliferation and differentiation were assessed by serial measurements of cell number and progesterone accumulation in the culture media, respectively. Granulosa cell apoptosis was evaluated using flow cytometry. Additional information about drug effects was obtained by immunoblotting to detect expression of proliferating cell nuclear antigen, p27(kip1) and the caspase-3 substrate poly(ADP-ribose) polymerase. The ERG channel antagonist MK-499 had no functional effects on cultured granulosa cells. However, the broad spectrum K(+) channel antagonist clofilium decreased, in a concentration-dependent fashion, the number of viable granulosa cells cultured, and these effects were associated with induction of apoptosis. All three I(Ks) antagonists (293B, L-735,821, and L-768,673) increased basal, but not FSH-enhanced progesterone accumulation on Day 1 after treatment without affecting the number of viable cells in culture, an effect that was blocked by pimozide. In contrast, CTX and MTX increased the number of viable cells in FSH-stimulated cultures on Day 3 after treatment without affecting progesterone output per cell. These data demonstrate that selective antagonism of granulosa cell K(+) channels with distinct molecular correlates, electrophysiological properties, and expression patterns can influence differential granulosa cell proliferation, steroidogenic capability, and apoptosis. Thus, K(+) channels may represent pharmacological targets for affecting Granulosa cell function and oocyte maturation, in vivo or in vitro. PMID- 12080004 TI - Bovine membrane-type 1 matrix metalloproteinase: molecular cloning and expression in the corpus luteum. AB - Matrix metalloproteinase-2 (MMP-2) is produced as a zymogen, which is subsequently activated by membrane-type 1 metalloproteinase (MT1-MMP). The objectives of the present study were to clone bovine MT1-MMP and to investigate its expression in the corpus luteum. Corpora lutea were harvested from nonlactating dairy cows on Days 4, 10, and 16 of the estrous cycle (Day 0 = estrus; n = 3 for each age). The bovine MT1-MMP cDNA contained an open reading frame of 1749 base pairs, which encoded a predicted protein of 582 amino acids. Northern blotting revealed no differences (P > 0.05) in MT1-MMP mRNA levels between any ages of corpora lutea. Western blotting demonstrated that two species of MT1-MMP, the latent form ( approximately 63 kDa) and the active form ( approximately 60 kDa), were present in corpora lutea throughout the estrous cycle. Active MT1-MMP was lower (P < 0.05) in early stages of the corpus luteum than the mid and late stages, where MMP-2 activity, as revealed by gelatin zymography, was also elevated. Furthermore, immunohistochemistry revealed that MT1-MMP was localized in endothelial, large luteal, and fibroblast cells of the corpus luteum at different stages. Taken together, the differential expression and localization of MT1-MMP in the corpus luteum suggest that it may have multiple functions throughout the course of the estrous cycle, including activation of pro-MMP-2. PMID- 12080005 TI - Differential regulation of pituitary hormone secretion and gene expression by thyrotropin-releasing hormone. A role for mitogen-activated protein kinase signaling cascade in rat pituitary GH3 cells. AB - We examined the possible involvement of mitogen-activated protein (MAP) kinase activation in the secretory process and gene expression of prolactin and growth hormone. Thyrotropin-releasing hormone (TRH) rapidly stimulated the secretion of both prolactin and growth hormone from GH3 cells. Secretion induced by TRH was not inhibited by 50 microM PD098059, but was completely inhibited by 1 microM wortmannin and 10 microM KN93, suggesting that MAP kinase does not mediate the secretory process. Stimulation of GH3 cells with TRH significantly increased the mRNA level of prolactin, whereas expression of growth hormone mRNA was largely attenuated. The increase in prolactin mRNA stimulated by TRH was inhibited by addition of PD098059, and the decrease in growth hormone mRNA was also inhibited by PD098059. Transfection of the cells with a pFC-MEKK vector (a constitutively active MAP kinase kinase kinase), significantly increased the synthesis of prolactin and decreased the synthesis of growth hormone. These data taken together indicate that MAP kinase mediates TRH-induced regulation of prolactin and growth hormone gene expression. Reporter gene assays showed that prolactin promoter activity was increased by TRH and was completely inhibited by addition of PD098059, but that the promoter activity of growth hormone was unchanged by TRH. These results suggest that TRH stimulates both prolactin and growth hormone secretion, but that the gene expressions of prolactin and growth hormone are differentially regulated by TRH and are mediated by different mechanisms. PMID- 12080007 TI - Progesterone blocks the estradiol-stimulated luteinizing hormone surge by disrupting activation in response to a stimulatory estradiol signal in the ewe. AB - The preovulatory surges of GnRH and LH are activated by increased concentrations of circulating estradiol, but ovulation is blocked when progesterone concentrations are elevated. Although it is has been shown that this action of progesterone is due to a central inhibition of the GnRH surge, the mechanisms that underlie the blockade of the GnRH surge are poorly understood. In this study we investigated whether progesterone can block the estradiol-dependent activation stage of the GnRH surge induction process, and thus prevent expression of the LH surge. The results demonstrated that exposure to progesterone for half or the full duration of the activation stage can prevent the stimulation of LH surges by estradiol (experiment 1), whereas exposure to progesterone midway though a period of estradiol exposure, which in itself is sufficient to activate the surge, did not block the LH surge (experiment 2). These results suggest that progesterone 1) disrupts activation of the surge induction system in response to a stimulatory estradiol signal and 2) does not compromise the ability of animals to respond to a stimulatory estradiol signal applied immediately after progesterone exposure. Because the disruptive effects of activated progesterone in response to estradiol are rapid but transient, it may be that progesterone directly interferes with the activation of estradiol-responsive neural systems to block the GnRH/LH surge. PMID- 12080006 TI - Signal transducer and activator of transcription 3 is expressed in the decidualized mesometrium of pregnancy and associates with the progesterone receptor through protein-protein interactions. AB - Progesterone is known to enhance epidermal growth factor (EGF)-mediated cellular responses by up-regulating EGF-receptor (EGF-R) expression. Ligand activation of EGF-R in turn has been shown to activate cytoplasmic stores of the STAT3 (signal transducer and activator of transcription) transcription factor, whereupon it translocates to the nucleus. The aim of this study was to examine the ontogeny of STAT3 protein expression in the decidualized mesometrium (i.e., decidua basalis) of the rat during pregnancy and its interactions with the progesterone progesterone receptor (PR) system. STAT3 was abundantly expressed in the cytosolic fraction of decidual homogenates throughout pregnancy; however, expression in the particulate fraction (assumed to reflect primarily nuclear accumulation) was reduced more than 75% on Days 12-17 than it was on Days 8 and 10. This pattern of expression parallels the decline in EGF-R and coincides with decidual regression. Treatment of pregnant rats with antiprogestin (RU486) in early pregnancy resulted in an 80% reduction in cytosolic abundance of STAT3 within 12 h, but it had no influence on STAT3 abundance in the particulate fraction. Immunoprecipitation of decidual lysates with PR or STAT3 antibodies resulted in coprecipitation of STAT3 and PR, respectively. These observations suggest that STAT3 expression is a prevalent feature of progesterone action, and that STAT3 and PR interactions represent a convergence of diverse signal transduction pathways in the decidualized mesometrium during pregnancy. PMID- 12080008 TI - Nuclei of nonviable ovine somatic cells develop into lambs after nuclear transplantation. AB - Here we report on the successful reprogramming of nuclei from somatic cells rendered nonviable by heat treatment. Granulosa cells from adult sheep were heated to nonphysiological temperatures (55 degrees C or 75 degrees C) before their nuclei were injected into enucleated metaphase II oocytes. Reprogramming was demonstrated by the capacity of the reconstructed embryos to develop to the blastocyst stage in vitro and into fetuses and viable offspring in suitable foster mothers. To our knowledge, this is the first report of cloned mammalian offspring originating from nonviable cells. In addition, our experiments show that heat-treating donor nuclei destabilizes higher-order features of chromatin (but leaves intact its nucleosomal organization) and results in a high proportion of reconstructed embryos developing to the blastocyst stage and beyond. PMID- 12080009 TI - Rabbit epididymal secretory proteins. I. Characterization and hormonal regulation. AB - Analyses of samples of luminal fluid from the rete testis, distal efferent ducts, and epididymal regions 2-5 and 8 revealed that 91% of the fluid leaving the testis is reabsorbed by the efferent ducts, 79% of the remainder is reabsorbed proximal to epididymal regions 4 and 5, and there is a net secretion of fluid into the duct caudally. There is a net reabsorption by the efferent ducts of 73% of the protein leaving the testis and then a net secretion along the epididymis. SDS-PAGE of the luminal fluids indicated that four new protein bands that were not present in blood appeared in the efferent ducts, 5 in epididymal regions 1-5, 6 in regions 6 and 7, and one in region 8. Two bands in samples from the efferent ducts were absent caudally, and one band present in region 7 was absent in region 8. The rates of incorporation of (35)S-methionine into minced duct in vitro varied among regions when expressed per milligram of wet weight of tissue (region 2-5 > region 7 > region 6 > region 1 > region 8 > ductuli efferentes), and orchidectomy had little effect on the rates. Incorporation into four proteins that were secreted in vitro (M(r) 38 000, 20 000, 15 000, and 13 000) was reduced or abolished by orchidectomy and restored by testosterone therapy. The secretion of three proteins (M(r) 52 000, 23 000, and 22 000) was reduced or abolished by orchidectomy and not restored by testosterone therapy. SDS-PAGE of detergent extracts of sperm indicated that five proteins were lost and nine were gained during epididymal transit. Seven of the proteins gained were about the same molecular weight as proteins secreted by the epididymis (M(r) 94 000, 52 000, 38 000, 36 000, 22 000, 20 000, and 13 000) and were analyzed using N-terminal amino acid microsequencing. PMID- 12080010 TI - Rabbit epididymal secretory proteins. II. Immunolocalization and sperm association of REP38. AB - Polyclonal antibody was used to partially characterize REP38, a major rabbit epididymal secretory protein. Western blot analyses and immunohistochemistry indicated that REP38 is only expressed in regions 5 and 6 of the epididymis (corpus epididy-midis) and is localized in the supranuclear region and microvilli of the principal cells in these regions. It was not expressed in other tissues of the body. In region 8 (cauda epididymidis), REP38 was detected in the luminal border and cytoplasm of scattered principal cells, indicating that it may be reabsorbed in this region. This protein accumulated on the sperm plasma membrane downstream of region 5 and was localized predominantly over the acrosomal and postacrosomal regions of the head and the middle piece. Although tightly bound to epididymal sperm, REP38 migrated to the equatorial segment under conditions in vivo that would promote capacitation. When tested in vitro, anti-REP38 IgG reduced the percentage of ova fertilized in a concentration-dependent manner, apparently by blocking sperm-egg fusion. PMID- 12080011 TI - Rabbit epididymal secretory proteins. III. Molecular cloning and characterization of the complementary DNA for REP38. AB - REP38 is a rabbit epididymal secretory protein of 38 kDa that has recently been shown to interact with spermatozoa. A rabbit epididymal cDNA expression library was screened with a polyclonal antibody raised against REP38. A single clone (REP38-c1) with an open reading frame encoding a polypeptide of 666 amino acids was obtained. Cleavage of a 22-amino acid N-terminal signal peptide revealed a mature protein with a theoretical molecular mass of 74.5 kDa. Northern blot analysis revealed the presence of two cross-hybridizing transcripts of approximately 1.3 and 2.5 kilobases that appear to result from alternative mRNA splicing. This finding may explain the discrepancies between the observed (38 kDa) and deduced molecular mass of REP38. Expression of both transcripts was epididymis specific and was detected only in regions 2-6. During development, the expression of REP38-c1 mRNA was initiated between 1 and 2 mo postnatum and therefore precedes the appearance of sperm within the lumen of the epididymis. These findings are in agreement with the immunohistochemical localization of the REP38 protein. Androgen deprivation induced by orchidectomy reduced REP38-c1 mRNA levels below the limit of detection, an effect that was reversed by administration of exogenous testosterone. Although REP38-c1 mRNA was detected only in the rabbit epididymis, database searches indicated homology with two rat testis specific cDNAs, KTT4 and odf2, which encode sperm outer dense fiber proteins. PMID- 12080012 TI - Epidermal growth factor enhances expression of connexin 43 protein in cultured porcine preantral follicles. AB - Connexin 43 (Cx43) and gap junctional coupling appear to play a critical role in early follicular development because absence of Cx43 disrupts progression of follicles beyond primary stages in transgenic mouse ovaries. Two experimental culture systems were used to determine whether epidermal growth factor (EGF) stimulates expression of Cx43 in early porcine follicular development. Ovarian explants were collected from 32- to 40-day-old gilts and cultured for 6 days on membrane inserts in Waymouth MB 752/1 medium supplemented with 0, 50, or 500 ng/ml mouse EGF. Western blot analysis demonstrated significant increases (P < 0.05) in relative amounts of Cx43 protein (both phosphorylated and nonphosphorylated) with 50 and 500 ng/ml of EGF as compared with control cultures. Preantral follicles were enzymatically isolated from 70- to 86-day-old gilts and cultured for 8 days in collagen matrices. Medium and EGF treatments were the same as previously described. Western blot analysis demonstrated a significant increase (P < 0.05) in relative amounts of Cx43 protein with 50 and 500 ng/ml of EGF as compared with control cultures. EGF increased expression of Cx43 protein in secondary preantral follicles in a dose-dependent manner, which suggests that EGF or similar growth factor molecules may modulate early folliculogenesis by stimulating expression of Cx43 gap junctions. PMID- 12080013 TI - Expression of cyclooxygenases 1 and 2 and prostaglandin E synthase in bovine endometrial tissue during the estrous cycle. AB - In ruminants, endometrial prostaglandin F(2alpha) (PGF(2alpha)) is responsible for luteolysis and prostaglandin E(2) (PGE(2)) is thought to be involved in maternal recognition of pregnancy. In the present study, healthy uteri were collected from cows at the abattoir, and days of the estrous cycle were determined macroscopically. The uteri were classified into seven groups as Days 1 3, 4-6, 7-9, 10-12, 13-15, 16-18, and 19-21 of the estrous cycle. Endometrial scrapings were collected. The expression of cyclooxygenase (COX)-1 and COX-2 mRNAs and proteins and PGE synthase (PGES) mRNA was analyzed by Northern and Western blot. There was no expression of COX-1, either mRNA or protein, on any day of the estrous cycle. In contrast, COX-2 mRNA and protein were expressed at low and high levels on Days 1-12 and 13-21 of the estrous cycle, respectively. The level of expression of PGES was moderate, low, and high on Days 1-3, 4-12, and 13-21 of the estrous cycle, respectively. There were significant correlations between COX-2 mRNA and protein levels and between COX-2 and PGES mRNA levels. COX 1 mRNA and protein are not expressed on any day of the estrous cycle, whereas COX 2 mRNA and protein and PGES mRNA are differentially expressed and regulated in bovine endometrium during the estrous cycle. COX-2, rather than COX-1, is the primary isoenzyme involved in the endometrial production of prostaglandins, and the COX-2 and PGES pathway is responsible for the endometrial production of PGE(2) in the bovine endometrium during the estrous cycle. PMID- 12080014 TI - High developmental competence of pig oocytes after meiotic inhibition with a specific M-phase promoting factor kinase inhibitor, butyrolactone I. AB - Butyrolactone I specifically inhibits M-phase promoting factor activation and prevents the resumption of meiosis. These experiments were conducted to examine effects of butyrolactone I on pig oocytes in a serum-free maturation system. The first experiment was conducted to determine the effect of butyrolactone I (0-100 microM) on nuclear maturation. At concentrations of > or =12.5 microM, germinal vesicle breakdown was prevented in >90% of the oocytes after 24 h of culture. In the second experiment, the kinetics of in vitro maturation of butyrolactone I treated oocytes was investigated. Oocytes were treated with 0 or 12.5 microM butyrolactone I and FSH for 20 h and then cultured with LH in the absence of butyrolactone I for another 24 h. Fewer butyrolactone I-treated oocytes reached MII stage at 36 h compared with controls (5.8% vs. 62.4%, P < 0.01). However, by 44 h, 83.4% of butyrolactone I-treated oocytes reached MII compared with 88.6% of controls. In the third experiment, butyrolactone I-treated oocytes were fertilized and cultured in vitro. No differences (P > 0.05) were found between controls and treated groups in cleavage rate, blastocyst rate, or mean number of cells per blastocyst. Effects of butyrolactone I on mitogen-activated protein kinase activation and localization of microfilaments and active mitochondria were examined by Western blot analysis and laser scanning confocal microscopy, respectively. The results suggested that although butyrolactone I reversibly inhibited germinal vesicle breakdown and mitogen-activated protein kinase activation, it did not affect mitochondrial and microfilament dynamics. Butyrolactone I is a potent inhibitor of nuclear maturation of porcine oocytes, and the inhibition is fully reversible. PMID- 12080016 TI - Direct inhibitory effect of progesterone on oxytocin-induced secretion of prostaglandin F(2alpha) from bovine endometrial tissue. AB - The effect of progesterone on oxytocin-induced secretion of prostaglandin (PG) F(2alpha) from bovine endometrial tissue explants was examined. Endometrial tissue from the late luteal phase were preincubated for 20 h in control medium. Explants were then treated for 6 h with control medium, oxytocin (10(-7) M), progesterone (10(-5) M), or both hormones. Oxytocin increased the medium concentration of 13,14-dihydro-15-keto-PGF(2alpha), whereas progesterone completely suppressed the stimulatory effect of oxytocin. In experiment 2, isolated endometrial epithelial cells were incubated with progesterone (10(-5) M), oxytocin (10(-7) M), and combinations of these hormones with or without actinomycin D (1 ng/ml). Only oxytocin stimulated secretion of PGF(2alpha), and this response was suppressed by progesterone. Oxytocin induced a rapid increase in intracellular concentrations of Ca(2+) detected within 1 min of exposure of epithelial cells from the same cows. Progesterone pretreatment diminished this response. In experiment 3, direct effects of progesterone (2 nM-20 microM) on binding of (3)H-oxytocin to the membrane preparation from epithelial cells were determined by saturation analysis. Oxytocin binding was suppressed by progesterone at every dosage tested. Progesterone is capable of suppressing the ability of oxytocin to induce endometrial secretion of PGF(2alpha). This effect appears to be mediated through a direct interference in the interaction of oxytocin with its own receptor. PMID- 12080015 TI - Placental endocrine disruption induced by cadmium: effects on P450 cholesterol side-chain cleavage and 3beta-hydroxysteroid dehydrogenase enzymes in cultured human trophoblasts. AB - We previously suggested that cadmium (Cd), an environmental toxicant and constituent of tobacco smoke, inhibits progesterone secretion in cultured human placental trophoblasts by inhibiting low-density lipoprotein receptor mRNA expression. In the current study, we investigated whether Cd also disrupts progesterone synthesis via P450 cholesterol side-chain cleavage (P450(scc)) and 3beta-hydroxysteroid dehydrogenase (3beta-HSD), enzymes that play important roles in placental steroidogenesis. Human cytotrophoblasts were purified by density gradient centrifugation and incubated in Dulbecco modified Eagle medium + 10% fetal bovine serum with 0, 5, 10, or 20 microM CdCl(2) for 96 h. Cells progressed to syncytiotrophoblastic maturity regardless of treatment. No differences (P > 0.05) in cell protein and lactate dehydrogenase activity were observed between untreated trophoblasts and those treated with CdCl(2). However, P450(scc) and 3beta-HSD mRNA transcript levels declined in a dose-dependent manner (P <0.05) in trophoblasts cocultured with 5, 10, or 20 microM CdCl(2). P450(scc) activity was similarly inhibited (P < 0.05) by CdCl(2) treatment, although 3beta-HSD activity was not significantly affected. Coculture with 8-bromo-cAMP enhanced progesterone secretion in untreated cultures but did not reverse the decline in progesterone secretion induced by CdCl(2) treatment. CdCl(2) failed to influence cAMP content in cultured cells. Collectively, results suggest that P450(scc) enzyme is another site at which Cd interferes with placental progesterone production. However, it is unlikely that an inhibition of cAMP is involved with the inhibition of progesterone biosynthesis by Cd in human trophoblasts. PMID- 12080017 TI - A role for protein kinase C during rat egg activation. AB - Upon sperm-egg interaction, an increase in intracellular calcium concentration ([Ca(2+)](i)) is observed. Several studies reported that cortical reaction (CR) can be triggered not only by a [Ca(2+)](i) rise but also by protein kinase C (PKC) activation. Because the CR is regarded as a Ca(2+)-dependent exocytotic process and because the calcium-dependent conventional PKCs (cPKC) alpha and beta II are considered as exocytosis mediators in various cell systems, we chose to study activation of the cPKC in the rat egg during in vivo fertilization and parthenogenetic activation. By using immunohistochemistry and confocal microscopy techniques, we demonstrated, for the first time, the activation of the cPKC alpha, beta I, and beta II during in vivo fertilization. All three isozymes examined presented translocation to the egg's plasma membrane as early as the sperm-binding stage. However, the kinetics of their translocation was not identical. Activation of cPKC alpha was obtained by the phorbol ester 12-O tetradecanoyl phorbol-13-acetate (TPA) or by 1-oleoyl-2-acetylglycerol (OAG) but not by the calcium ionophore ionomycin. PKC alpha translocation was first detected 5-10 min after exposure to TPA and reached a maximum at 20 min, whereas in eggs activated by OAG, translocation of PKC alpha was observed almost immediately and reached a maximum within 5 min. These results suggest that, although [Ca(2+)](i) elevation on its own does not activate PKC alpha, it may accelerate OAG-induced PKC alpha activation. We also demonstrate a successful inhibition of the CR by a myristoylated PKC pseudosubstrate (myrPKCPsi), a specific PKC inhibitor. Our study suggests that exocytosis can be triggered independently either by a [Ca(2+)](i) rise or by PKC. PMID- 12080018 TI - Stage-specific expression of the mitochondrial germ cell epitope PG2 during postnatal differentiation of rabbit germ cells. AB - Structural and biochemical differentiation of germ cell mitochondria is supposed to determine the fate and integrity of mitochondria in the early embryo. Immunofluorescent labeling of the primordial germ cell epitope 2 (PG2), which is associated with the outer mitochondrial membrane and is germ cell specific from the time of germ cell segregation during gastrulation, was used to elucidate biochemical characteristics of mitochondrial differentiation leading to a functional gamete. The PG2 epitope is found in both mitotic and meiotic male and female postnatal germ cells, but PG2 expression ceases transiently in initial stages of meiosis, i.e., in the female during early stages of follicle formation and in the male during prespermatogenesis and initial phases of spermatogenesis. Because the PG2 epitope is detectable in germ cells at the time when structurally immature mitochondria are present, we speculate that PG2 immunoreactivity closely mirrors the progress of mitochondrial differentiation during gametogenesis. PMID- 12080019 TI - Retinoic acid mediates transcriptional repression of ovine follicle-stimulating hormone receptor gene via a pleiotropic nuclear receptor response element. AB - The FSH receptor (FSHR) and retinoid receptors are critical regulators of gonadal function. Unlike the latter, the FSH receptors are expressed exclusively in ovarian granulosa and testicular Sertoli cells in a developmental fashion. Toward understanding the nature of various transcription factors that direct a tissue- and stage-specific expression of the FSHR gene, we have studied FP4, one of the two footprinting regions (FP3 and FP4) mapped at -241 to -269 and -284 to -303, respectively, upstream of the transcription start site of the ovine FSHR gene. Gel mobility shift assays with FP4 probe revealed two sequence-specific DNA protein complexes in the presence of nuclear extracts from two immortal gonadal cell lines. Antibody supershift assays demonstrated that retinoic acid receptor (RAR) was involved in the complex 1 whereas steroidogenic factor-1 (SF-1) was present in the complex 2. Mutation studies revealed that DNA binding sites for RAR and SF-1 were overlapping each other within a 19-base pair length of nucleotide sequence of FP4, and a mutation in the half RAR binding site seriously affected SF-1 binding. Reporter assays showed that FP4 conferred SF-1 transactivation as well as RAR-mediated, ligand-dependent repression. Overexpression of SF-1 in a transformed Sertoli cell line partially overcame RAR mediated suppression. For the first time, our studies reveal a direct retinoid modulation of the gonadotropin receptor promoter and suggest a mechanism by which activators and repressors compete for composite elements providing antagonistic pathways that could modulate the expression of FSHR. PMID- 12080021 TI - Altered meiotic regulation in oocytes from diabetic mice. AB - In the present study, we have utilized a streptozotocin-induced diabetic mouse model to examine how the diabetic condition and different glucose concentrations affect several parameters of reproductive physiology. We report that oocyte maturation is altered under all experimental conditions examined. In cumulus cell enclosed oocytes (CEO) from diabetic mice, spontaneous maturation was accelerated but the FSH-mediated delay of spontaneous maturation was suppressed. Higher glucose levels in the culture medium suppressed spontaneous maturation but did not influence the transient arrest mediated by FSH. Meiotic arrest in CEO by hypoxanthine and dibutyryl cAMP (dbcAMP) was less effective at higher glucose concentrations. In addition, both FSH-induced maturation in vitro and hCG-induced maturation in vivo were reduced by the diabetic condition. The ovulation rate was lowered by about 50% in diabetic mice and fewer ovulated ova had reached metaphase II. Despite the decreased number of ova at metaphase II, in vitro cultures showed the oocytes were capable of completing meiotic maturation at control levels. Insulin treatment reversed the detrimental effects of diabetes on meiotic induction, ovulation, and completion of meiotic maturation. Cultures of pronuclear-staged embryos confirmed a negative effect of diabetes and hyperglycemia on development to the blastocyst stage. These data suggest that defects in meiotic regulation brought about by the diabetic condition are due to decreased communication between the somatic and germ cell compartments, and it is concluded that such conditions may contribute to postfertilization developmental abnormalities. PMID- 12080020 TI - Role of sperm surface arylsulfatase A in mouse sperm-zona pellucida binding. AB - We have previously described the zonae pellucidae (ZP) binding ability of a pig sperm surface protein, P68. Our recent results on peptide sequencing of 3 P68 tryptic peptides and molecular cloning of pig testis arylsulfatase A (AS-A) revealed the identity of P68 as AS-A. In this report, we demonstrate the presence of AS-A on the mouse sperm surface and its role in ZP binding. Using anti-AS-A antibody, we have shown by immunoblotting that AS-A was present in a Triton X-100 extract of mouse sperm. The presence of AS-A on the sperm plasma membrane was conclusively demonstrated by indirect immunofluorescence, immunogold electron microscopy, and AS-A's desulfation activity on live mouse sperm. The AS-A remained on the head surface of in vivo capacitated sperm, as revealed by positive immunofluorescent staining of oviductal/uterine sperm. Significantly, the role of mouse sperm surface AS-A on ZP binding was demonstrated by dose dependent decreases of sperm-ZP binding on sperm pretreatment with anti-AS-A IgG/Fab. Furthermore, Alexa-430 conjugated AS-A bound to mouse ZP of unfertilized eggs but not to fertilized ones, and this level of binding increased and approached saturation with increasing Alexa-430 AS-A concentrations. Moreover, in vivo fertilization was markedly decreased when mouse sperm pretreated with anti AS-A IgG were artificially inseminated into females. All of these results designated a new function for AS-A in mouse gamete interaction. PMID- 12080022 TI - Three forms of gonadotropin-releasing hormone, including a novel form, in a basal salmonid, Coregonus clupeaformis. AB - Multiple forms of GnRH within individual brains may have different functions. However, some vertebrates such as salmonids continue to reproduce even though they have lost or do not express 1 of the 3 forms of GnRH found in most other teleosts. We examined a basal salmonid, lake whitefish, to determine the mechanism by which a reduction in the number of GnRH forms occurs. We identified for the first time 3 distinct GnRHs in a salmonid. One form is novel and is designated whitefish GnRH. The primary structure is pGlu-His-Trp-Ser-Tyr-Gly-Met Asn-Pro-Gly-NH(2). HPLC and RIA were used for purification followed by Edman degradation for sequence determination. Mass spectroscopy was used to confirm the sequence and amidation of the peptide. The other 2 forms, salmon GnRH and chicken GnRH-II, are identical to the 2 forms found in salmon, which evolved later than whitefish. Synthetic whitefish GnRH is biologically active, as it increased mRNA expression of growth hormone and the alpha-subunit for LH and thyroid-stimulating hormone in dispersed fish pituitary cells. Our data support the hypothesis that the ancestral salmonid had a third GnRH form when the genome doubled (tetraploidization), but the third form was lost later in some salmonids due to chromosomal rearrangements. We suggest that the salmon GnRH form compensated for the loss of the third form. PMID- 12080023 TI - Differential expression of activator protein-1 transcription factors in pregnant rat myometrium. AB - While the AP-1 (activator protein-1) genes c-fos and c-jun have been implicated in the expression of myometrial genes associated with the onset of labor, there are no data concerning the role of other members of this family of transcription factors. To address this issue, we defined the expression and hormonal regulation of AP-1 genes in the rat myometrium during pregnancy and labor. Tissue was collected on Days 12, 15, 17, 19, 21, 22, and 23 (labor) and 1 day postpartum. Expression of c-fos, fosB, fra-1, fra-2, and junB was low during early gestation, with a 5- to 10-fold increase on Day 23 during labor, and returned to low levels 1 day postpartum. In contrast, the levels of c-jun and junD remained relatively constant throughout gestation. Administration of progesterone (P4; 16 mg/kg s.c./day) beginning on Day 20 (to maintain elevated plasma P4 levels) prevented the onset of labor and blocked the expected rise in c-fos, fosB, fra-1, fra-2, and junB expression on Day 23. In contrast, administration of the progesterone receptor antagonist RU486 (10 mg/kg s.c.) on Day 19 induced preterm labor and a premature increase in mRNA levels of c-fos, fra-1, fra-2, and junB. In unilaterally pregnant rats, stretch imposed by the growing fetus was found to increase the expression of c-fos, fosB, fra-1, fra-2, and junB only in the gravid horn on the day of labor. These data raise the possibility that AP-1 transcription factors integrate endocrine and mechanical signals, leading to myometrial gene expression required for uterine remodeling and the initiation of labor. PMID- 12080024 TI - Comparative testis morphometry and seminiferous epithelium cycle length in donkeys and mules. AB - The mule (Equus mulus mulus) is a sterile hybrid domestic animal that results from the breeding of a male donkey (Equus asinus) to a female horse (Equus caballus). Usually, spermatogenesis in mules does not advance beyond spermatocytes. In the present study, we performed a comparative and more accurate morphometric and functional investigation of the testis in donkeys and mules. Due to the smaller testis size, lower seminiferous tubule volume density, and fewer germ cells, the total length of seminiferous tubules in mules was significantly smaller than in donkeys. However, the percentage of seminiferous tubules containing germ cells (spermatogonia and spermatocytes) in mules was approximately 95%. The total number of Sertoli cells per testis observed in donkeys and mules was very similar. However, the total number of Leydig cells in mules was approximately 70% lower than in donkeys. At least in part, this difference was probably related to the lower number of germ cells present in mule seminiferous tubules. Although spermatogenesis in mules did not advance beyond secondary spermatocytes/newly formed round spermatids, germ cell associations in the seminiferous epithelium and pachytene spermatocytes nuclear volume in donkeys and mules were similar. The duration of spermatogenesis was estimated using intratesticular injections of tritiated thymidine. Each spermatogenic cycle in donkeys lasted 10.5 days. A similar value was found in mules ( approximately 10.1 days). Considering that the entire spermatogenic process takes approximately 4.5 cycles to be completed, its total duration in donkeys was estimated to last 47.2 days. The results found for mules suggest that the mechanisms involved in the determination of testis structure and function are probably originated from donkeys. Also, the data found for mules suggest that their seminiferous tubules are able to sustain complete spermatogenesis. In this regard, this species is a potential model for transplants of germ cells originated from donkeys and horses or other large animals. PMID- 12080025 TI - In vitro fertilization of in vitro-matured equine oocytes: effect of maturation medium, duration of maturation, and sperm calcium ionophore treatment, and comparison with rates of fertilization in vivo after oviductal transfer. AB - Three experiments were conducted to evaluate the effect of oocyte and sperm treatments on rates of in vitro fertilization (IVF) in the horse and to determine the capacity of in vitro-matured horse oocytes to be fertilized in vivo. There was no effect of duration of oocyte maturation (24 vs. 42 h) or calcium ionophore concentration during sperm capacitation (3 microM vs. 7.14 microM) on in vitro fertilization rates. Oocytes matured in 100% follicular fluid had significantly higher fertilization (13% to 24%) than did oocytes matured in maturation medium or in 20% follicular fluid (0% to 12%; P < 0.05). There was no significant difference in fertilization rate among 3 sperm treatments utilizing 7.14 microM calcium ionophore (12% to 21%). Of in vitro-matured oocytes recovered 40-44 h after transfer to the oviducts of inseminated mares, 77% showed normal fertilization (2 pronuclei to normal cleavage). Cleavage to 2 or more cells was seen in 22% of oocytes matured in follicular fluid and 63% of oocytes matured in maturation medium; this difference was significant (P < 0.05). We conclude that in vitro-matured horse oocytes are capable of being fertilized at high rates in the appropriate environment and that in vitro maturation of oocytes in follicular fluid increases fertilization rate in vitro but reduces embryo development after fertilization in vivo. Further work is needed to determine the optimum environment for sperm capacitation and IVF in the horse. PMID- 12080026 TI - Relationship of serum testosterone concentrations to mate preferences in rams. AB - This study examined systemic testosterone concentrations in rams that were classified according to their sexual behavior and partner preference as either female-oriented (FOR), male-oriented (MOR), or asexual (NOR). For this purpose, we measured testosterone concentrations under three separate conditions: in conscious rams during the nonbreeding season (June) and breeding season (November), and in anesthetized rams during the breeding season. Basal testosterone concentrations in conscious rams were not different among the three groups (P > 0.05) in either season. However, when rams were anesthetized, mean systemic concentrations of testosterone in FORs (mean +/- SEM, 13.9 +/- 7.4 ng/ml serum) were greater (P < 0.05) than in NORs (0.9 +/- 0.1 ng/ml), but not in MORs (2.2 +/- 6.2 ng/ml), whereas testosterone concentrations were not different between MORs and NORs (P > 0.05). Concentrations of testosterone in the spermatic vein of FORs (127 +/- 66 ng/ml) were greater (P < 0.05) than in MORs (41 +/- 10 ng/ml) and NORs (19 +/- 7 ng/ml). Serum LH concentrations were not different. Cortisol was higher (P < 0.05) in anesthetized MORs (25.1 +/- 4.2 ng/ml) and NORs (27.2 +/- 4.4 ng/ml) than in FORs (10.9 +/- 1.8 ng/ml). These results demonstrate that circulating testosterone concentrations are related to sexual behavior only when rams are bled under anesthesia. Thus, differences in basal androgen concentrations in adulthood cannot be responsible for expression of male-oriented preferences or low libido in sheep. Instead, functional differences must exist between the brains of rams that differ in sexual preference expression. PMID- 12080027 TI - Sperm volume regulation: maturational changes in fertile and infertile transgenic mice and association with kinematics and tail angulation. AB - Laser light scatter analyzed by flow cytometry was used to monitor the volume of viable maturing murine spermatozoa. Upon release, dispersion, and dilution, epididymal sperm from fertile heterozygous c-ros knockout mice were smallest in the cauda region and largest in the corpus region. Cauda sperm from both infertile homozygous c-ros knockout and GPX5-Tag2 transgenic mice were abnormally large. When incubated, corpus and cauda sperm from normal mice became slightly enlarged and later returned to a smaller size. This suggests an immediate swelling due to high intracellular osmolality, which triggers a regulatory volume decrease (RVD) that results in a net volume reduction. Normal caput sperm increased in size continuously and became larger than the more mature sperm, indicating a lack of RVD. The ion-channel blocker quinine induced dose-dependent size increases in normal cauda sperm but not in caput sperm. Dose-dependent quinine action on mature sperm also included induction of tail angulation, and suppression of straight-line velocity and linearity. The kinematic effects were more sensitive, with a quicker onset, but they diminished with time in contrast to tail angulation, which intensified. These results suggest that kinematic changes are an early phenomenon of swelling, which gradually accumulates at the cytoplasmic droplet to cause flagellar angulation. Disruption of the epididymal maturation of sperm volume regulation capacity would hinder the transport of sperm in the female tract, and may thereby explain infertility under certain conditions, but may also provide a novel approach to male contraception. PMID- 12080028 TI - Localization of a proteolytic enzyme within the efferent and deferent duct epithelial cells of the turkey (Meleagris gallopavo) using immunohistochemistry. AB - Turkey seminal plasma contains a serine protease found to be distinct from the spermatozoal acrosin. However, the origin and biological roles of this enzyme are unknown. Our experimental objective was to identify the cellular source of this protease within the male reproductive tract. The enzyme was isolated from seminal plasma using benzamidine-Sepharose 6B chromatography. A synthetic substrate, Nalpha-benzoyl-DL-arginine p-nitroanilide, was used to detect fractions containing the enzyme. The affinity chromatography technique yielded a 150-fold increase in amidase activity. Analysis of the protease by SDS-PAGE revealed two protein bands with relative molecular masses of 37 000 and 61 000. Proteolytic activity was detected within the smaller band as evidenced by casein digestion. Further analysis of the purified protein revealed that the smaller protein band was glycosylated. To determine the cellular source of the protease, a panel of mouse monoclonal antibodies was then developed against the purified protease, and used in immunohistochemistry. Frozen tissue sections from the liver, testis, epididymal region, and deferent duct were fixed in 4% (w/v) paraformaldehyde, permeabilized with 0.2% (v/v) (octylphenoxy)polyethoxyethanol followed by routine immunohistochemistry procedures. Monoclonal antibodies did not bind to tissue sections from either the liver or testis, or to blood plasma proteins. Both the distal portion of the efferent duct and the deferent duct were immunoreactive. We concluded that the protease found in turkey seminal plasma is concentrated to the distal efferent duct and the deferent duct epithelial cells. PMID- 12080029 TI - Quantitative analysis of gene expression in preimplantation mouse embryos using green fluorescent protein reporter. AB - We have developed a method to monitor noninvasively, quantitatively, and in real time transcription in living preimplantation mouse embryos by measuring expression of a short half-life form of enhanced green fluorescent protein (EGFP) following microinjection of a plasmid-borne EGFP reporter gene. A standard curve was established by injecting known amounts of recombinant green fluorescent protein, and transcriptional activity was then determined by interpolating the amount of fluorescence in the DNA-injected embryos. This approach permitted multiple measurements in single embryos with no significant detrimental effect on embryonic development as long as light exposure was brief (<30 sec) and no more than two measurements were made each day. This method should facilitate analysis of the regulation of gene expression in preimplantation embryos; in particular, during the maternal-to-zygotic transition, and in other species in which limited numbers of embryos are available. PMID- 12080030 TI - Separation of motile populations of spermatozoa prior to freezing is beneficial for subsequent fertilization in vitro: a study with various mouse strains. AB - Success with in vitro fertilization (IVF) using inbred strains of mice varies considerably and appears to be related to the proportion of motile spermatozoa present in epididymal sperm samples of different strains. In this study, motile spermatozoa were separated from the original samples using a column of Sephadex G25. IVF rates were compared between separated and nonseparated samples of epididymal spermatozoa before and after cryopreservation. Oocytes and spermatozoa were obtained from FVB, DBA/2, C57BL/6J, and BALB/c inbred mice; and from F1 (C57BL/6J;ts DBA/2) hybrid mice, and isogenic gametes were used for IVF. These strains of mice were chosen because of their common use in transgenesis and mutagenesis studies. Dulbecco PBS was used for sperm separation on Sephadex, 18% raffinose, and 3% skim milk for cryopreservation; T6 medium for IVF; and mKSOM(AA) for embryo culture. There was a marked improvement in the rate of fertilization using fresh spermatozoa after motile spermatozoa were separated in C57BL/6J and BALB/c strains (92% vs. 58%, 79% vs. 44%) but no differences were found in fertilization rates between separated and nonseparated spermatozoa in F1, FVB, and DBA/2 strains (99% vs. 83%, 95% vs. 93%, 86% vs. 87%, respectively). After cryopreservation, higher rates of fertilization were obtained with separated motile samples in all strains; the greatest improvements were obtained with spermatozoa from C57BL/6J and BALB/c strains (40% vs. 16% and 51% vs. 14% for separated and nonseparated spermatozoa, respectively). No differences were found between the proportions of 14.5-day fetuses developing from embryos derived from separated and nonseparated spermatozoa with or without cryopreservation (33% to 46%). In conclusion, the fertility of frozen-thawed mouse epididymal spermatozoa improves significantly when highly motile populations of spermatozoa are separated for freezing. PMID- 12080031 TI - Purification and characterization of relaxin from the tammar wallaby (Macropus eugenii): bioactivity and expression in the corpus luteum. AB - The objective of this study was to isolate and purify prorelaxin or mature relaxin from the tammar wallaby corpus luteum (CL), determine their structure and bioactivity, and test the hypothesis that enzymatic cleavage of prorelaxin occurs in late gestation. Tammar relaxin peptides were extracted from pooled corpora lutea of late pregnant tammars using a combination of HPLC methods, and they were identified using Western blotting with a human (H2) relaxin antisera and matrix assisted laser desorption ionization time of flight mass spectrometry. Although no prorelaxin was identified, multiple 6-kDa peptides were detected, which corresponded to the predicted mature tammar relaxin amino acid sequence, with an A chain of 24 amino acids, and different B chain lengths of 28, 29, 30, and 32 amino acids. Tammar relaxin bound with high affinity to rat cortical relaxin receptors and stimulated cAMP production in the human monocytic cell line, THP-1, which expresses the relaxin receptor. Analysis of individual CL indicated that equivalent amounts of mature relaxin peptides were present throughout gestation and also in unmated tammars at equivalent stages of the luteal phase in the nonpregnant cycle. Immunoreactive relaxin was localized specifically to the luteal cells of the CL and the intensity of immunostaining did not vary between gestational stages. These data show that the CL of both pregnant and unmated tammar wallabies produces mature relaxin and suggests that relaxin expression in this species is not influenced by the conceptus. Moreover, the presence of mature relaxin throughout gestation implies that prohormone cleavage is not limited to the later stages of pregnancy PMID- 12080032 TI - Regulation of the human sperm tyrosine kinase c-yes. Activation by cyclic adenosine 3',5'-monophosphate and inhibition by Ca(2+). AB - During the process of capacitation, spermatozoa go through a whole set of signaling cascade events in order to become fully competent at fertilizing the egg. An increase in sperm protein tyrosine phosphorylation has been described during this final maturational event in different animal species as well as in humans. Although the phosphotyrosine content of sperm protein is modulated by cAMP, Ca(2+), BSA, oxygen derivatives, and cholesterol, no protein tyrosine kinase (PTK) nor the phosphotyrosine protein phosphatase (PTPase) directly involved in the control of the phosphotyrosine content of sperm protein has been identified. Therefore, the goal of the present study was to identify the tyrosine kinases putatively responsible for the increases in sperm protein phosphotyrosine content. In the present study, we show that the src-related tyrosine kinase c-yes is present in the head of human spermatozoa in both membranes and Triton X-100 insoluble extracts. Our hypothesis was that c-yes is a tyrosine kinase responsible for at least some of the capacitation-induced increase in protein tyrosine phosphorylation. When spermatozoa were previously incubated in the presence of 3-isobutyl-1-methylxanthine or 1,2-bis-(o-aminophenoxy)-ethane N,N,N',N'-tetraacetic acid, treatments known to increase the phosphotyrosine content of human sperm proteins, an increase in the kinase activity of immunoprecipitated yes was measured using enolase as a substrate. These results suggest that cAMP activates while Ca(2+) inhibits human sperm c-yes kinase activity. PMID- 12080033 TI - Selected proteins of "prostasome-like particles" from epididymal cauda fluid are transferred to epididymal caput spermatozoa in bull. AB - During epididymal transit, spermatozoa acquire selected proteins secreted by epithelial cells. We recently showed that P25b, a protein with predictive properties for bull fertility, is transferred from prostasome-like particles present in the cauda epididymal fluid (PLPCd) to the sperm surface. To further characterize the interactions between PLPCd and epididymal spermatozoa, PLPCd were prepared by ultracentrifugation of bull epididymal fluid, then surface exposed proteins were biotinylated and coincubated in different conditions with caput epididymal spermatozoa. Western blot analysis revealed that only selected proteins are transferred from PLPCd to spermatozoa. MALDI-TOF analysis revealed that these transferred proteins are closely related. The pattern of distribution of the PLPCd transferred varied from one sperm cell to the other, with a bias toward the acrosomal cap. This transfer appeared to be temperature sensitive, being more efficient at 32-37 degrees C than at 22 degrees C. Transfer of PLPCd proteins to spermatozoa was also pH dependant, the optimal pH for transfer being 6.0-6.5. The effect of divalent cations on PLPCd protein transfer to caput spermatozoa was investigated. Whereas Mg(2+) and Ca(2+) have no effect on the amount of proteins remaining associated with spermatozoa following coincubation, Zn(2+) had a beneficial effect. These results are discussed with regard to the function of PLPCd in epididymal sperm maturation. PMID- 12080034 TI - Organic cation/carnitine transporter, OCTN2, is differentially expressed in the adult rat epididymis. AB - L-Carnitine must be transported against a substantial concentration gradient across the epididymal epithelium to achieve high intraluminal levels, approximately 50 mM in the cauda. Recently, an organic cation transporter, OCTN2, was cloned from rat intestinal epithelium and shown to transport L-carnitine in a sodium-dependent manner. To test the hypothesis that OCTN2 was present in the epididymis, primers were designed based on the published OCTN2 mRNA sequence. A 1.9-kilobase OCTN2 cDNA from rat epididymis was amplified by reverse transcription polymerase chain reaction (RT-PCR) and cloned. Northern analysis demonstrated the presence of OCTN2 transcripts in the epididymis, with highest expression in the distal caput and corpus. To localize the protein, an antibody raised against a carboxy-terminal peptide of OCTN2 was produced in rabbits and used for Western blot analysis and immunohistochemistry. The antibody recognized a band of approximately 65 kDa in Western blots using epididymal lysates. Immunohistochemical studies demonstrate that OCTN2 is present in the basolateral membrane of epithelial cells in the distal caput, corpus, and proximal cauda epididymides. In conclusion, OCTN2 is present in the rat epididymis in a region dependent manner and is likely to be responsible for the transport of L-carnitine into the cells of the epididymal epithelium. PMID- 12080035 TI - Effect of uterine blood flow occlusion on shear stress-mediated nitric oxide production and endothelial nitric oxide synthase expression during ovine pregnancy. AB - During normal pregnancy, uterine blood flow (UBF) is increased in association with elevations of endothelial nitric oxide (NO) production and endothelial nitric oxide synthase (eNOS) expression. Shear stress increases endothelial derived NO production to reduce vasomotor tone. We hypothesized that decreasing in vivo UBF, and thus shear stress, will decrease NO and/or eNOS levels. In this experiment, one of the main uterine arteries of chronically instrumented late pregnant sheep (125 +/- 1 days' gestation [mean +/- SEM]; n = 15) was occluded for 24 h. Cardiovascular parameters (systemic and uterine arterial pressure, heart rate [HR], and ipsilateral and contralateral UBF) and NO(2)/NO(3) (NO(x)) levels were evaluated. Although UBF measured using Transonic flow probes was reduced unilaterally 41.5% +/- 2.1%, uterine perfusion pressure only fell 12.2% +/- 4.5%. Systemic arterial blood pressure and HR were unaltered. Using radioactive microspheres, ipsilateral UBF was reduced approximately 28% during occlusion. The redistribution of UBF to other reproductive tissues suggests that collateral circulation develops in response to occlusion. Systemic arterial and uterine venous NO(x) levels were reduced 22.1% +/- 6.7% and 22.6% +/- 7.6%, respectively, during occlusion. Treatment with microspheres produced an unexpected initial ( approximately 2.5 h) increase in systemic arterial and uterine venous NO(x) levels by 116% +/- 30% and 97% +/- 49%, respectively. Despite a decline in NO(x) levels after 6 h, no significant differences versus preocclusion NO(x) levels were detected by 24 h of occlusion in this experimental group. In contrast, NO(x), UBF, and uterine perfusion pressure levels unexpectedly failed to return to baseline values following release of occlusion. No differences in uterine artery eNOS expression were demonstrated by Western analysis from occlusion. Thus, our data suggest that shear stress may mediate in vivo vasomotor tone via production of NO(x). PMID- 12080036 TI - Production of cloned cattle from in vitro systems. AB - The pregnancy initiation and maintenance rates of nuclear transfer embryos produced from several bovine cell types were measured to determine which cell types produced healthy calves and had growth characteristics that would allow for genetic manipulation. Considerable variability between cell types from one animal and the same cell type from different animals was observed. In general, cultured fetal cells performed better with respect to pregnancy initiation and calving than adult cells with the exception of cumulous cells, which produced the highest overall pregnancy and calving rates. The cell type that combined relatively high pregnancy initiation and calving rates with growth characteristics that allowed for extended proliferation in culture were fetal genital ridge (GR) cells. Cultured GR cells used in nuclear transfer and embryo transfer initiated pregnancies in 40% of recipient heifers (197), and of all recipients that received nuclear transfer embryos, 9% produced live calves. Cultured GR cells doubled as many as 85 times overall and up to 75 times after dilution to single cell culture. A comparison between transfected and nontransfected cells showed that transfected cells had lower pregnancy initiation (22% versus 32%) and calving (3.4% versus 8.9%) rates. PMID- 12080037 TI - Ontogeny of cloned cattle to lactation. AB - Central to the success of large animal cloning is the production of healthy animals that can provide products for human health, food, and other animal agriculture applications. We report development of cloned cattle derived from 34 genetically unique, nonembryonic cell lines using nuclear transfer performed between 1 January 1998 and 29 February 2000. Nearly 25% (535/2170) of the recipients receiving reconstructed embryos initiated pregnancy. Overall, 19.8% (106/535) of the initiated pregnancies resulted in live births, while 77% (82/106) of these cattle clones remain healthy and productive today. Although a wide variation in birth weight of clone calves was observed, their growth rates, reproductive performance, and lactation characteristics are similar to that found in noncloned dairy cattle. Our data represent the most comprehensive information on cattle derived from nuclear transfer procedures and indicate that this emerging reproductive technology offers unique opportunities to meet critical needs in both human health care and agriculture. PMID- 12080038 TI - Capacitation induces cyclic adenosine 3',5'-monophosphate-dependent, but apoptosis-unrelated, exposure of aminophospholipids at the apical head plasma membrane of boar sperm cells. AB - The capacitating agent bicarbonate/CO(2) has been shown to induce profound changes in the architecture and dynamics within the sperm's plasma membrane lipid bilayer via a cAMP-dependent protein phosphorylation signaling pathway. Here we have investigated the effect of bicarbonate on surface exposure of endogenous aminophospholipids in boar spermatozoa, detecting phosphatidylserine (PS) with fluorescein-conjugated annexin V and phosphatidylethanolamine (PE) with fluorescein-conjugated streptavidin/biotinylated Ro-09-0198. Flow cytometric analyses revealed that incubation with 15 mM bicarbonate induced 30%-70% of live acrosome-intact cells to expose PE very rapidly; this exposure was closely related to a decrease in lipid packing order as detected by enhanced binding of merocyanine 540. PS exposure was detectable in the same proportion of cells, though its expression was slower. Confocal microscopy revealed that exposure of aminophospholipids in intact cells was restricted to the anterior acrosomal region of the head plasma membrane. Aminophospholipid exposure, merocyanine stainability, and a subsequent migration of cholesterol to the apical region of the head plasma membrane, were all under the control of the cAMP-dependent protein phosphorylation pathway. The close coupling of decreased lipid packing order with exposure of PE led us to conclude that bicarbonate was inducing phospholipid scrambling (i.e., collapse of asymmetric transverse distribution), and that the scrambling was a prerequisite for cholesterol relocation. There was no evidence whatever that the bicarbonate-induced scrambling was an apoptotic process. It was not accompanied by major loss of viability or by DNA degeneration or by loss of mitochondrial function, and it could not be blocked by the broad specificity caspase inhibitors zVAD-fmk and BocD-fmk. In the absence of bicarbonate, scrambling could not be induced by the apoptotic agents UV, staurosporine, or cycloheximide. Bicarbonate-induced phospholipid scrambling thus appears to be an important and early physiological event in the capacitation process. PMID- 12080039 TI - Differential expression of microsomal prostaglandin e synthase at implantation sites and in decidual cells of mouse uterus. AB - Prostaglandin E(2) (PGE(2)) is considered important for blastocyst spacing, implantation, and decidualization in the rodent uterus. PGE synthase (PGES) catalyzes the isomerization of PGH(2) to PGE(2). There are two isoforms of PGES, microsomal PGES (mPGES) and cytosolic PGES (cPGES). However, the expression and regulation of mPGES in the mammalian uterus during early pregnancy are still unknown. The aim of this study was to investigate the differential expression of mPGES in mouse uterus during early pregnancy and its regulation under different conditions by in situ hybridization and immunohistochemistry. Microsomal PGES expression in the preimplantation mouse embryos was also performed by reverse transcription polymerase chain reaction (RT-PCR). Expression of mPGES mRNA and protein was at a basal level in the luminal epithelium from Day 1 to Day 4 of pregnancy. However, mPGES mRNA and protein were highly expressed in the stroma immediately surrounding the blastocyst but not in the luminal epithelium on Day 5 of pregnancy. Microsomal PGES mRNA and protein were not detected in the pseudopregnant uterus from Day 1 to Day 5. During delayed implantation, mPGES mRNA and protein were also not detected in the uterus. Once delayed implantation was terminated by estrogen treatment and embryo implantation initiated, both mPGES mRNA and protein were induced to express in the stroma immediately surrounding the blastocyst, which was similar to the expression pattern on Day 5 of pregnancy. From Day 6 to Day 8 of pregnancy, the signals for mPGES mRNA and protein were strongly detected in the decidualized cells. Microsomal PGES mRNA and protein were also highly expressed in the artificially decidualized cells but not in the control horn. Microsomal PGES mRNA was detected in the oocytes and all the stages of preimplantation embryos. The strong mPGES expression in the implantation site and decidual cells suggests that mPGES might play an important role during implantation and more importantly in decidualization. PMID- 12080040 TI - The anti-estrogenic effect of all-trans-retinoic acid on the breast cancer cell line MCF-7 is dependent on HES-1 expression. AB - All-trans-retinoic acid has been shown to have an antiproliferative effect in the estrogen receptor alpha-positive breast cancer cell line MCF-7. The mechanism of this effect is not well understood. We have previously shown that 17beta estradiol down-regulates the basic helix-loop-helix factor Hairy and Enhancer of Split homologue-1 in MCF-7 and T47D cells (Strom, A., Arai, N., Leers, J., and Gustafsson, J. A. (2000) Oncogene 19, 5951-5953) and that this down-regulation is essential for proliferation in response to 17beta-estradiol. Treatment of the same cells with all-trans-retinoic acid prevented 17beta-estradiol-mediated down regulation of the factor. The antiproliferative effect of all-trans-retinoic acid correlated well with the prevention of Hairy and Enhancer of Split homologue-1 down-regulation. Increasing concentrations of all-trans-retinoic acid, in the range of 1-1000 nm, produced a dose-dependent inhibition of proliferation and prevented 17beta-estradiol-mediated down-regulation of Hairy and Enhancer of Split homologue-1. By using a receptor-specific ligand we were able to show that the retinoic acid receptor alpha is important for regulation of the Hairy and Enhancer of Split homologue-1. Expression of a dominant negative form of Hairy and Enhancer of Split homologue-1 in MCF-7 cells abolished the growth-inhibitory effect of all-trans-retinoic acid in these cells. This finding indicates that Hairy and Enhancer of Split homologue-1 is a mediator of the antiproliferative effect of all-trans-retinoic acid in estrogen receptor alpha-positive breast cancer cell lines. PMID- 12080041 TI - Characterization and channel coupling of the P2Y(12) nucleotide receptor of brain capillary endothelial cells. AB - Rat brain capillary endothelial (B10) cells express an unidentified nucleotide receptor linked to adenylyl cyclase inhibition. We show that this receptor in B10 cells is identical in sequence to the P2Y(12) ADP receptor ("P2Y(T)") of platelets. When expressed heterologously, 2-methylthio-ADP (2-MeSADP; EC(50), 2 nm), ADP, and adenosine 5'-O-(2-thio)diphosphate were agonists of cAMP decrease, and 2-propylthio-D-beta,gamma-difluoromethylene-ATP was a competitive antagonist (K(B), 28 nm), as in platelets. However, 2-methylthio-ATP (2-MeSATP) (EC(50), 0.4 nm), ATP (1.9 microm), and 2-chloro-ATP (190 nm), antagonists in the platelet, were also agonists. 2-MeSADP activated (EC(50), 0.1 nm) GIRK1/GIRK2 inward rectifier K(+) channels when co-expressed with P2Y(12) receptors in sympathetic neurons. Surprisingly, P2Y(1) receptors expressed likewise gave that response; however, a full inactivation followed, absent with P2Y(12) receptors. A new P2Y(12)-mediated transduction was found, the closing of native N-type Ca(2+) channels; again both 2-MeSATP and 2-MeSADP are agonists (EC(50), 0.04 and 0.1 nm, respectively). That action, like their cAMP response, was pertussis toxin sensitive. The Ca(2+) channel inhibition and K(+) channel activation are mediated by beta gamma subunit release from a heterotrimeric G-protein. G alpha subunit types in B10 cells were also identified. The presence in the brain capillary endothelial cell of the P2Y(12) receptor is a significant extension of its functional range. PMID- 12080042 TI - Oligomeric states of the detergent-solubilized human serum paraoxonase (PON1). AB - Human plasma paraoxonase (HuPON1) is a high density lipoprotein (HDL)-bound enzyme exhibiting antiatherogenic properties. The molecular basis for the binding specificity of HuPON1 to HDL has not been established. Isolation of HuPON1 from HDL requires the use of detergents. We have determined the activity, dispersity, and oligomeric states of HuPON1 in solutions containing mild detergents using nondenaturing electrophoresis, size exclusion chromatography, and cross-linking. HuPON1 was active whatever its oligomeric state. In nonmicellar solutions, HuPON1 was polydisperse. In contrast, HuPON1 exhibited apparent homogeneity in micellar solutions, except with CHAPS. The enzyme apparent hydrodynamic radius varied with the type of detergent and protein concentration. In C(12)E(8) micellar solutions, from sedimentation velocity, equilibrium analytical ultracentrifugation, and radioactive detergent binding, HuPON1 was described as monomers and dimers in equilibrium. A decrease of the detergent concentration shifted this equilibrium toward the formation of dimers. About 100 detergent molecules were associated per monomer and dimer. The assembly of amphiphilic molecules, phospholipids in vivo, in sufficiently large aggregates could be a prerequisite for anchoring of HuPON1 and then allowing stabilization of the enzyme activity. Changes of HDL size and shape could strongly affect the binding affinity and stability of HuPON1 and result in reduced antioxidative capacity of the lipoprotein. PMID- 12080043 TI - p73 Interacts with c-Myc to regulate Y-box-binding protein-1 expression. AB - YB-1 is a member of the cold shock domain family of proteins that is important for signaling DNA damage and cell proliferation. YB-1 is induced by DNA damage and can also recognize cisplatin-modified DNA. In this study we observed a 6-fold increase in the steady-state level of YB-1 mRNA in response to cisplatin exposure in cells of the human cancer cell line KB. We present evidence from cotransfection experiments for a critical role of c-Myc and p73 in the transactivation of the YB-1 promoter. p73 transactivated the YB-1 promoter in experiments with Saos-2 cells, which express c-Myc, but not with HO15.19 cells, which lack c-Myc. In turn, c-Myc transactivated an intact YB-1 promoter but not a YB-1 promoter with a mutant E-box, indicating that the E-box is necessary for the response of the promoter to cisplatin. We also found that p73 interacts with c Myc in vitro and in vivo. Using deletion mutants we showed that the DNA-binding domain of p73 and the C-terminal region of c-Myc are required for the interaction. Furthermore, p73 stimulated the interaction of Max with c-Myc and promoted binding of the c-Myc-Max complex to its target DNA. Our data suggest that p73 stimulates the transcription of the YB-1 promoter by enhancing recruitment of the c-Myc-Max complex to the E-box. PMID- 12080044 TI - The promyelocytic leukemia protein represses A20-mediated transcription. AB - The promyelocytic leukemia (PML) protein is a tumor suppressor that is disrupted by the chromosomal translocation t(15;17), a consistent cytogenetic feature of acute promyelocytic leukemia. A role of PML in multiple pathways of apoptosis was conclusively demonstrated using PML(-/-) animal and cell culture models. In a previous study, we found that PML sensitizes tumor necrosis factor-induced apoptosis in tumor necrosis factor (TNF)-resistant U2OS cells. This finding helped to explain the mechanism of PML-induced apoptosis. The zinc finger protein A20 is a target gene of NF kappa B inducible by TNF alpha, and it is a potent inhibitor of TNF-induced apoptosis. In the this study, we demonstrated that PML is a transcriptional repressor of the A20 promoter and that PML represses A20 expression induced by TNF alpha. We showed that PML inhibits A20 transactivation through the NF kappa B site by interfering with its binding to the promoter. We also showed that stable overexpression of A20 inhibits apoptosis and caspase activation induced by PML/TNF alpha. The results of this study suggest that A20 is a downstream target of PML-induced apoptosis and supports a role of A20 in modulating cell death induced by PML/TNF alpha in TNF-resistant cells. PMID- 12080045 TI - Characterization of a heparan sulfate octasaccharide that binds to herpes simplex virus type 1 glycoprotein D. AB - Herpes simplex virus type 1 utilizes cell surface heparan sulfate as receptors to infect target cells. The unique heparan sulfate saccharide sequence offers the binding site for viral envelope proteins and plays critical roles in assisting viral infections. A specific 3-O-sulfated heparan sulfate is known to facilitate the entry of herpes simplex virus 1 into cells. The 3-O-sulfated heparan sulfate is generated by the heparan sulfate d-glucosaminyl-3-O-sulfotransferase isoform 3 (3-OST-3), and it provides binding sites for viral glycoprotein D (gD). Here, we report the purification and structural characterization of an oligosaccharide that binds to gD. The isolated gD-binding site is an octasaccharide, and has a binding affinity to gD around 18 microm, as determined by affinity coelectrophoresis. The octasaccharide was prepared and purified from a heparan sulfate oligosaccharide library that was modified by purified 3-OST-3 enzyme. The molecular mass of the isolated octasaccharide was determined using both nanoelectrospray ionization mass spectrometry and matrix-assisted laser desorption/ionization mass spectrometry. The results from the sequence analysis suggest that the structure of the octasaccharide is a heptasulfated octasaccharide. The proposed structure of the octasaccharide is DeltaUA-GlcNS IdoUA2S-GlcNAc-UA2S-GlcNS-IdoUA2S-GlcNH(2)3S6S. Given that the binding of 3-O sulfated heparan sulfate to gD can mediate viral entry, our results provide structural information about heparan sulfate-assisted viral entry. PMID- 12080046 TI - Characterization of Saccharomyces cerevisiae acyl-protein thioesterase 1, the enzyme responsible for G protein alpha subunit deacylation in vivo. AB - Thioacylation is a reversible lipid modification of proteins that plays a role in the regulation of signal transduction. Acyl-protein thioesterase 1 (APT1) was identified as an enzyme capable of deacylating some thioacylated proteins in vitro. Saccharomyces cerevisiae open reading frame YLR118c encodes an enzyme homologous to Rattus norvegicus APT1. We demonstrate that the catalytic activity of the protein encoded by the yeast open reading frame is similar to that of rat APT1, and we designate the protein S. cerevisiae Apt1p. Yeasts bearing a disruption of the APT1 gene lack significant biochemically detectable acyl protein thioesterase activity. They also fail to deacylate Gpa1p, the yeast G alpha subunit, in metabolic radiolabeling studies. We conclude that native APT1 is the enzyme responsible for G alpha subunit deacylation in S. cerevisiae and presumably other eukaryotes as well. PMID- 12080047 TI - The adenosine 2b receptor is recruited to the plasma membrane and associates with E3KARP and Ezrin upon agonist stimulation. AB - We have previously shown that adenosine is formed in the intestinal lumen during active inflammation from neutrophil-derived 5'-AMP. Acting through the adenosine A2b receptor (A2bR), the luminally derived adenosine induces vectorial chloride secretion and a polarized secretion of interleukin-6 to the intestinal lumen. Although some G protein-coupled receptors interact with anchoring or signaling molecules, not much is known in this critical area for the A2bR. We used the model intestinal epithelial cell line, T84, and Caco2-BBE cells stably transfected with GFP-A2b receptor to study the intestinal A2bR. The A2bR is present in both the apical and basolateral membranes of intestinal epithelia. Apical or basolateral stimulation of the A2bR induces recruitment of the receptor to the plasma membrane and caveolar fractions. The A2bR co-immunoprecipitates with E3KARP and ezrin upon agonist stimulation. Ezrin interacts with E3KARP and PKA and the interaction between ezrin and E3KARP is enhanced by agonist stimulation. Our data suggest that the A2bR is recruited to the plasma membrane upon apical or basolateral agonist stimulation and interacts with E3KARP and ezrin. We speculate that such an interaction may not only anchor the A2bR to the plasma membrane but may also function to stabilize the receptor in a signaling complex in the plasma membrane. PMID- 12080048 TI - Xenopus ribosomal RNA gene intergenic spacer elements conferring transcriptional enhancement and nucleolar dominance-like competition in oocytes. AB - Repeated within the intergenic spacers that separate adjacent ribosomal RNA (rRNA) genes in Xenopus laevis are several distinct sequence elements. These include transcription terminators, "region 0" repeats, "region 1" repeats, duplicated spacer promoters, and 42-bp enhancer elements that are embedded within 60 or 81-bp repeats. All have been reported to stimulate RNA polymerase I transcription from an adjacent gene promoter. A greater number of 42-bp enhancers/gene have been suggested to explain the preferential transcription of X. laevis rRNA genes in X. laevis x Xenopus borealis hybrids, an epigenetic phenomenon known as nucleolar dominance. However, the possible contribution of regions 0/1 and/or spacer promoters to the preferential transcription of X. laevis (over X. borealis) rRNA genes has never been tested directly. In this study, we systematically tested the various intergenic spacer elements for their contributions to promoter strength and nucleolar dominance-like competition in oocytes. In disagreement with a previous report, region 0 and region 1 repeats do not have significant enhancer activity, nor do they play a discernible role in X. laevis-X. borealis rRNA gene competition. Minigenes containing X. laevis spacer sequences are only dominant over minigenes having complete X. borealis spacers if a spacer promoter is located upstream of the 42-bp enhancers; X. laevis enhancers alone are not sufficient. These results provide additional evidence that spacer promoters together with adjacent enhancers form a functional activating unit in Xenopus oocytes. PMID- 12080049 TI - A conserved serine juxtaposed to the pseudosubstrate site of type I cGMP dependent protein kinase contributes strongly to autoinhibition and lower cGMP affinity. AB - Serines 64 and 79 are homologous residues that are juxtaposed to the autoinhibitory pseudosubstrate site in cGMP-dependent protein kinase type Ialpha and type Ibeta (PKG-Ialpha and PKG-Ibeta), respectively. Autophosphorylation of this residue is associated with activation of type I PKGs. To determine the role of this conserved serine, point mutations have been made in PKG-Ialpha (S64A, S64T, S64D, and S64N) and PKG-Ibeta (S79A). In wild-type PKG-Ialpha, basal kinase activity ratio (-cGMP/+cGMP) is 0.11, autophosphorylation increases this ratio 3 fold, and the K(a) and K(D) values for cGMP are 127 and 36 nm, respectively. S64A PKG-Ialpha basal kinase activity ratio increases 2-fold, cGMP binding affinity increases approximately 10-fold in both K(a) and K(D), and activation by autophosphorylation is slight. S64D and S64N mutants are nearly constitutively active in the absence of cGMP, cGMP binding affinity in each increases 18-fold, and autophosphorylation does not affect the kinase activity of these mutants. Mutation of the homologous site in PKG-Ibeta (S79A) increases the basal kinase activity ratio 2-fold and cGMP binding affinity 5-fold over that of wild-type PKG Ibeta. The combined results demonstrate that a conserved serine juxtaposed to the pseudosubstrate site in type I PKGs contributes importantly to enzyme function by increasing autoinhibition and decreasing cGMP binding affinity. PMID- 12080050 TI - The requirement of H1 histones for a heterodimeric nuclear import receptor. AB - After synthesis in the cytoplasm, H1 histones are imported into the nucleus through an energy-dependent process that can be mediated by an importin beta importin 7 (Impbeta-Imp7) heterodimer. H1 histones contain two structurally different types of nuclear localization signals (NLS). The first type of NLS resides within the unstructured C-terminal domain and is rich in basic amino acids. In contrast, the highly conserved central domain of the H1 histone contains comparatively few basic amino acids but also represents a functional NLS. The competence for the nuclear import of this globular domain seems to be based on its secondary structure. Here, we show that the Impbeta-Imp7 heterodimer is the only receptor for H1 import. Furthermore, we identified the import receptors mediating the in vitro transport of different NLS of the H1 histone. Using the digitonin-permeabilized cell import assay we show that Impbeta is the most efficient import receptor for the globular domain of H1 histones, whereas the heterodimer of Impbeta and Imp7 is the functional receptor for the entire C terminal domain. However, short fragments of the C-terminal domain are imported in vitro by at least four different importins, which resembles the import pathway of ribosomal proteins and core histones. In addition, we show that heterodimerization of Impbeta with Imp7 is absolutely necessary for their proper function as an import receptor for H1 histones. These findings point to a chaperone-like function of the heterodimeric complex in addition to its function as an import receptor. It appears that the Impbeta-Imp7 heterodimer is specialized for NLS consisting of extended basic domains. PMID- 12080051 TI - Cloning and characterization of a protein kinase A anchoring protein (AKAP) related protein that interacts with and regulates sphingosine kinase 1 activity. AB - Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid metabolite that has novel dual actions. S1P is the ligand for a family of G protein-coupled receptors known as S1PRs that mediate various physiological functions. Growth factors rapidly activate sphingosine kinase type 1 (SPHK1) resulting in phosphorylation of sphingosine to form S1P, which plays important roles in cell growth regulation and protection from apoptosis. However, little is known of the mechanism(s) by which SPHK activity is regulated. Using a yeast two-hybrid screening approach, we cloned a 3-kb cDNA encoding a SPHK1-interacting protein (SKIP). BLAST analysis revealed that SKIP corresponded to the C-terminal region of a larger ( approximately 7 kb) cDNA that encoded a protein with a high degree of similarity to a family of protein kinase A anchor proteins (AKAP). In confirmation of the yeast two-hybrid assay, glutathione S-transferase (GST)-SPHK1 specifically pulled down SKIP, whereas GST did not. Moreover, immunoprecipitation of in vitro translated SPHK1 and SKIP revealed that SKIP and SPHK1 are tightly associated. Furthermore, SKIP overexpression in NIH 3T3 fibroblasts reduced SPHK1 activity and interfered with its biological functions. The apoptotic-sparing effect of SPHK1 against serum deprivation was reduced when co-transfected with SKIP. In addition, SPHK1-enhanced cell proliferation was also abolished by SKIP, with a corresponding decrease in activation of ERK. Taken together, these results indicate that SKIP is a novel protein likely to play a regulatory role in the modulation of SPHK1 activity. PMID- 12080052 TI - Human mitochondrial thioredoxin. Involvement in mitochondrial membrane potential and cell death. AB - Thioredoxins (Trx) are a class of small multifunctional redox-active proteins found in all organisms. Recently, we reported the cloning of a mitochondrial thioredoxin, Trx2, from rat heart. To investigate the biological role of Trx2 we have isolated the human homologue, hTrx2, and generated HEK-293 cells overexpressing Trx2 (HEK-Trx2). Here, we show that HEK-Trx2 cells are more resistant toward etoposide. In addition, HEK-Trx2 are more sensitive toward rotenone, an inhibitor of complex I of the respiratory chain. Finally, overexpression of Trx2 confers an increase in mitochondrial membrane potential, DeltaPsi(m). Treatment with oligomycin could both reverse the effect of rotenone and decrease the membrane potential suggesting that Trx2 interferes with the activity of ATP synthase. Taken together, these results suggest that Trx2 interacts with specific components of the mitochondrial respiratory chain and plays an important role in the regulation of the mitochondrial membrane potential. PMID- 12080053 TI - Specific inhibition of ICAM-1 expression mediated by gene targeting with Triplex forming oligonucleotides. AB - Selected sequences in the DNA double helix can be specifically recognized by oligonucleotides via hydrogen bonding interactions. The resulting triple helix can modulate DNA metabolism and especially interfere with transcription in a gene specific manner. To explore the potential of triplex-forming oligonucleotides (TFOs) as gene repressors, a TFO was designed to target a 16-bp sequence within the third intron of the human intercellular-adhesion molecule-1 (ICAM-1) gene, which plays a key role in initiating inflammation. TFO binding to its ICAM-1 target sequence was characterized in vitro and also demonstrated in cell nuclei with the set-up of a novel magnetic capture assay, which represents a general experimental approach to the detection of specific TFO binding and to the determination of the accessibility of a given genomic DNA locus. In a human keratinocyte cell line (A431), we observed that: (i) the ICAM-1 target sequence in the chromatin context within the nuclei is still available for triplex formation and (ii) TFO inhibits sequence and gene-specific interferon-gamma induced ICAM-1 surface expression. Collectively, the data demonstrate effective and specific inhibition of ICAM-1 expression by TFO treatment and support the view that triplex-mediated gene targeting might be a valuable technique for anti inflammatory or anticancer strategies. PMID- 12080054 TI - RAP80, a novel nuclear protein that interacts with the retinoid-related testis associated receptor. AB - In this study, we describe the characterization of a novel nuclear protein, referred to as RAP80. The RAP80 cDNA was cloned from a human testis cDNA library and encodes a 719-amino acid protein containing two potential CX(2)CX(11)HX(3)C type zinc finger motifs at its carboxyl-terminal region. Analysis of its genomic structure revealed that the RAP80 gene covers more than 90 kb and consists of 15 exons and 14 introns. Fluorescence in situ hybridization mapped the RAP80 gene to human chromosome 5q35. RAP80 mRNA is expressed in many human tissues, but its expression is particularly high in testis. In situ hybridization showed that RAP80 is highly expressed in germ cells of mouse testis but is not differentially regulated during spermatogenesis. Confocal microscopy showed that RAP80 is localized to the nucleus, where it is distributed in a speckled pattern. Deletion analysis showed that a bipartite nuclear localization signal at the amino terminus is important in mediating nuclear transport of RAP80. Monohybrid analysis showed that RAP80 might function as an active repressor of transcription. Mammalian two-hybrid analysis demonstrated that RAP80 was able to interact with the retinoid-related testis-associated receptor (RTR), an orphan receptor that has been implicated in the control of embryonic development and spermatogenesis. Pull-down analysis showed that RAP80 and RTR physically interact in vitro. Deletion and point mutation analyses revealed that part of the hinge domain of RTR is required for this interaction. RAP80 is able to inhibit the interaction of RTR with the co-repressor N-CoR likely by competing with N-CoR for RTR binding. Our results suggest that RAP80 may be functioning as a modulator of RTR signaling. PMID- 12080056 TI - Structural elements that govern the substrate specificity of the clot-dissolving enzyme plasmin. AB - There is remarkable homology between the core structures of plasmin, a fibrin clot-degrading enzyme, and factor D, a complement-activating enzyme, despite markedly different biological functions. We postulated that sequence divergence in the loop structures between these two enzymes mediated the unique substrate and inhibitor interactions of plasmin. Recombinant microplasminogens chimerized with factor D sequences at loops 3, 5, and 7 were cleaved by the plasminogen activator urokinase and developed titratable active sites. Chimerization abolished functional interactions with the plasminogen activator streptokinase but did not block complex formation. The microplasmin chimeras showed enhanced resistance (k(i) decreased up to two to three times) to inactivation of microplasmin by alpha(2)-antiplasmin. Microplasmin chimerization had minimal ( approximately 2 fold) effects on the catalytic efficiency for cleavage of small substrates and did not alter the cleavage of fibrin. However, microplasmin and the microplasmin chimeras showed enhanced abilities to degrade fibrin in plasma clots suspended in human plasma. These studies indicate that loop regions of the protease domain of plasmin are important for interactions with substrates, regulatory molecules, and inhibitors. Because modification of these regions affected substrate and inhibitor interactions, loop chimerization may hold promise for improving the clot dissolving properties of this enzyme. PMID- 12080055 TI - Sit4 is required for proper modulation of the biological functions mediated by Pkc1 and the cell integrity pathway in Saccharomyces cerevisiae. AB - Maintenance of cellular integrity in Saccharomyces cerevisiae is carried out by the activation of the protein kinase C-mediated mitogen-activated protein kinase (PKC1-MAPK) pathway. Here we report that correct down-regulation of both basal and induced activity of the PKC1-MAPK pathway requires the SIT4 function. Sit4 is a protein phosphatase also required for a proper cell cycle progression. We present evidence demonstrating that the G(1) to S delay in the cell cycle, which occurs as a consequence of the absence of Sit4, is mediated by up-regulation of Pkc1 activity. Sit4 operates downstream of the plasma membrane sensors Mid2, Wsc1, and Wsc2 and upstream of Pkc1. Sit4 affects all known biological functions involving Pkc1, namely Mpk1 activity and cell wall integrity, actin cytoskeleton organization, and ribosomal gene transcription. PMID- 12080057 TI - p52 Mediates XPB function within the transcription/repair factor TFIIH. AB - To further our understanding of the transcription/DNA repair factor TFIIH, we investigated the role of its p52 subunit in TFIIH function. Using a completely reconstituted in vitro transcription or nucleotide excision repair (NER) system, we show that deletion of the C-terminal region of p52 results in a dramatic reduction of TFIIH NER and transcription activities. This mutation prevents promoter opening and has no effect on the other enzymatic activities of TFIIH. Moreover, we demonstrate that intact p52 is needed to anchor the XPB helicase within TFIIH, providing an explanation for the transcription and NER defects observed with the mutant p52. We show that these two subunits physically interact and map domains involved in the interface. Taken together, our results show that the p52/Tfb2 subunit of TFIIH regulates the function of XPB through pair-wise interactions as described previously for p44 and XPD. PMID- 12080058 TI - Vitreoscilla hemoglobin binds to subunit I of cytochrome bo ubiquinol oxidases. AB - The bacterium, Vitreoscilla, can induce the synthesis of a homodimeric hemoglobin under hypoxic conditions. Expression of VHb in heterologous bacteria often enhances growth and increases yields of recombinant proteins and production of antibiotics, especially under oxygen-limiting conditions. There is evidence that VHb interacts with bacterial respiratory membranes and cytochrome bo proteoliposomes. We have examined whether there are binding sites for VHb on the cytochrome, using the yeast two-hybrid system with VHb as the bait and testing every Vitreoscilla cytochrome bo subunit as well as the soluble domains of subunits I and II. A significant interaction was observed only between VHb and intact subunit I. We further examined whether there are binding sites for VHb on cytochrome bo from Escherichia coli and Pseudomonas aeruginosa, two organisms in which stimulatory effects of VHb have been observed. Again, in both cases a significant interaction was observed only between VHb and subunit I. Because subunit I contains the binuclear center where oxygen is reduced to water, these data support the function proposed for VHb of providing oxygen directly to the terminal oxidase; it may also explain its positive effects in Vitreoscilla as well as in heterologous organisms. PMID- 12080059 TI - Crystal structures at atomic resolution reveal the novel concept of "electron harvesting" as a role for the small tetraheme cytochrome c. AB - The genus Shewanella produces a unique small tetraheme cytochrome c that is implicated in the iron oxide respiration pathway. It is similar in heme content and redox potential to the well known cytochromes c(3) but related in structure to the cytochrome c domain of soluble fumarate reductases from Shewanella sp. We report the crystal structure of the small tetraheme cytochrome c from Shewanella oneidensis MR-1 in two crystal forms and two redox states. The overall fold and heme core are surprisingly different from the soluble fumarate reductase structures. The high resolution obtained for an oxidized orthorhombic crystal (0.97 A) revealed several flexible regions. Comparison of the six monomers in the oxidized monoclinic space group (1.55 A) indicates flexibility in the C-terminal region containing heme IV. The reduced orthorhombic crystal structure (1.02 A) revealed subtle differences in the position of several residues, resulting in decreased solvent accessibility of hemes and the withdrawal of a positive charge from the molecular surface. The packing between monomers indicates that intermolecular electron transfer between any heme pair is possible. This suggests there is no unique site of electron transfer on the surface of the protein and that electron transfer partners may interact with any of the hemes, a process termed "electron-harvesting." This optimizes the efficiency of intermolecular electron transfer by maximizing chances of productive collision with redox partners. PMID- 12080060 TI - The ATP:Co(I)rrinoid adenosyltransferase (CobA) enzyme of Salmonella enterica requires the 2'-OH group of ATP for function and yields inorganic triphosphate as its reaction byproduct. AB - The specificity of the ATP:corrinoid adenosyltransferase (CobA) enzyme of Salmonella enterica serovar Typhimurium LT2 for its nucleotide substrate was tested using ATP analogs and alternative nucleotide donors. The enzyme showed broad specificity for the nucleotide base and required the 2'-OH group of the ribosyl moiety of ATP for activity. (31)P NMR spectroscopy was used to identify inorganic triphosphate (PPP(i)) as the byproduct of the reaction catalyzed by the CobA enzyme. Cleavage of triphosphate into pyrophosphate and orthophosphate did not occur, indicating that triphosphate cleavage was not required for release of the adenosylcorrinoid product. Triphosphate was a strong inhibitor of the reaction, with 85% of CobA activity lost when the ATP/PPP(i) ratio present in the reaction mixture was 1:2.5. PMID- 12080061 TI - Identification of a tankyrase-binding motif shared by IRAP, TAB182, and human TRF1 but not mouse TRF1. NuMA contains this RXXPDG motif and is a novel tankyrase partner. AB - Tankyrase-1 and -2 are closely related poly(ADP-ribose) polymerases that use an ankyrin-repeat domain to bind diverse proteins, including TRF (telomere-repeat binding factor)-1, IRAP (insulin-responsive aminopeptidase), and TAB182 (182-kDa tankyrase-binding protein). TRF1 binding allows tankyrase to regulate telomere dynamics in human cells, whereas IRAP binding presumably allows tankyrase to regulate the targeting of IRAP. The mechanism by which tankyrase binds to diverse proteins has not been investigated. Herein we describe a novel RXXPDG motif shared by IRAP, TAB182, and human TRF1 that mediates their binding to tankyrases. Interestingly, mouse TRF1 lacks this motif and thus does not bind either tankyrase-1 or -2. Using the ankyrin domain of tankyrase as a bait in a yeast two hybrid screen, we also found the RXXPDG motif in six candidate tankyrase partners, including the nuclear/mitotic apparatus protein (NuMA). We verified NuMA as an RXXPDG-mediated partner of tankyrase and suggest that this interaction contributes to the known colocalization of tankyrase and NuMA at mitotic spindle poles. PMID- 12080062 TI - Two novel transcripts for human endothelin B receptor produced by RNA editing/alternative splicing from a single gene. AB - Hirschsprung disease is a heterogeneous genetic disorder, causative genes of which include the endothelin B receptor (ETB). To investigate the mutations of ETB in Hirschsprung disease, expression of the ETB gene in lymphoblastoid cells from patients and normal healthy adults was examined, and novel mutant transcripts were found. The mutant ETB gene transcripts lacked a 134-bp nucleotide sequence corresponding to exon 5, and some also contained a substitution from A to G at position 950 in exon 4, resulting in an amino acid substitution from glutamine (Q) to arginine (R). This substitution was suspected to be the result of RNA editing because it was not present in the genomic sequence. Transfection experiments using ETB minigenes containing the editing site with or without the gene for double-strand RNA deaminases (ADAR1 and ADAR2) revealed that the deaminases were involved in RNA editing. Furthermore, a c-Myc tagged mutant ETB protein was not detected by Western blot analysis. The present results show that the mutant ETB transcripts were novel splice variants, which might not be translated, or that the products translated from splice variants might be quickly degraded, presumably because of their instability. The preferential production of this null function ETB by RNA editing/splicing could be involved in the etiology of some cases of Hirschsprung disease. PMID- 12080063 TI - Reduced flavins promote oxidative DNA damage in non-respiring Escherichia coli by delivering electrons to intracellular free iron. AB - When cells are exposed to external H(2)O(2), the H(2)O(2) rapidly diffuses inside and oxidizes ferrous iron, thereby forming hydroxyl radicals that damage DNA. Thus the process of oxidative DNA damage requires only H(2)O(2), free iron, and an as-yet unidentified electron donor that reduces ferric iron to the ferrous state. Previous work showed that H(2)O(2) kills Escherichia coli especially rapidly when respiration is inhibited either by cyanide or by genetic defects in respiratory enzymes. In this study we established that these respiratory blocks accelerate the rate of DNA damage. The respiratory blocks did not substantially affect the amounts of intracellular free iron or H(2)O(2), indicating that that they accelerated damage because they increased the availability of the electron donor. The goal of this work was to identify that donor. As expected, the respiratory inhibitors caused a large increase in the amount of intracellular NADH. However, NADH itself was a poor reductant of free iron in vitro. This suggests that in non-respiring cells electrons are transferred from NADH to another carrier that directly reduces the iron. Genetic manipulations of the amounts of intracellular glutathione, NADPH, alpha-ketoacids, ferredoxin, and thioredoxin indicated that none of these was the direct electron donor. However, cells were protected from cyanide-stimulated DNA damage if they lacked flavin reductase, an enzyme that transfers electrons from NADH to free FAD. The K(m) value of this enzyme for NADH is much higher than the usual intracellular NADH concentration, which explains why its flux increased when NADH levels rose during respiratory inhibition. Flavins that were reduced by purified flavin reductase rapidly transferred electrons to free iron and drove a DNA-damaging Fenton system in vitro. Thus the rate of oxidative DNA damage can be limited by the rate at which electron donors reduce free iron, and reduced flavins become the predominant donors in E. coli when respiration is blocked. It remains unclear whether flavins or other reductants drive Fenton chemistry in respiring cells. PMID- 12080065 TI - Primates highly responsive to dietary cholesterol up-regulate hepatic ACAT2, and less responsive primates do not. AB - The role of liver acyl-CoA:cholesterol acyltransferase 2 (ACAT2), earlier shown to be the principal ACAT enzyme within primate hepatocytes, as a regulator of the hypercholesterolemia induced by dietary cholesterol was studied. At the end of low and high cholesterol diet periods, liver biopsies were taken from cynomolgus monkeys, a species highly responsive to dietary cholesterol, and less responsive African green monkeys. Liver cholesterol and cholesteryl ester concentrations were highest in cynomolgus monkeys fed cholesterol, despite the fact that in order to induce equivalent hypercholesterolemia, dietary cholesterol levels were 50% lower than was fed to green monkeys. Hepatic cholesteryl oleate secretion rate, measured during liver perfusion as an indicator of ACAT activity, was significantly higher in cynomolgus monkeys. Liver microsomal ACAT activity was 2 3-fold higher in cynomolgus monkeys than in green monkeys. The responses of ACAT2 were compared with those of ACAT1 that is found primarily in Kupffer cells. ACAT2 protein mass was significantly correlated to microsomal total ACAT activity in both species; ACAT1 mass was less well correlated. Dietary cholesterol induced a significant 3-fold increase of ACAT2 protein mass in cynomolgus monkeys, a much greater increase than was found for mRNA abundance; neither ACAT2 mRNA nor protein was diet-responsive in green monkeys. In cynomolgus monkeys but not in green monkeys, liver free cholesterol concentrations were elevated when cholesterol was fed and were correlated with ACAT2 protein levels. The data suggest a mechanism whereby the elevation of hepatic free cholesterol concentrations by dietary cholesterol, seen only in cynomolgus monkeys, resulted in higher ACAT2 protein levels in hepatocytes, either through increased production or stabilization of the protein. Regulation of ACAT2 gene transcription was not a factor. PMID- 12080064 TI - Chromatin structural analyses of the mouse Igkappa gene locus reveal new hypersensitive sites specifying a transcriptional silencer and enhancer. AB - To identify new regulatory elements within the mouse Igkappa locus, we have mapped DNase I hypersensitive sites (HSs) in the chromatin of B cell lines arrested at different stages of differentiation. We have focused on two regions encompassing 50 kilobases suspected to contain new regulatory elements based on our previous high level expression results with yeast artificial chromosome-based mouse Igkappa transgenes. This approach has revealed a cluster of HSs within the 18-kilobase intervening sequence, which we cloned and sequenced in its entirety, between the Vkappa gene closest to the Jkappa region. These HSs exhibit pro/pre-B cell-specific transcriptional silencing of a Vkappa gene promoter in transient transfection assays. We also identified a plasmacytoma cell-specific HS in the far downstream region of the locus, which in analogous transient transfection assays proved to be a powerful transcriptional enhancer. Deletional analyses reveal that for each element multiple DNA segments cooperate to achieve either silencing or enhancement. The enhancer sequence is conserved in the human Igkappa gene locus, including NF-kappaB and E-box sites that are important for the activity. In summary, our results pinpoint the locations of presumptive regulatory elements for future knockout studies to define their functional roles in the native locus. PMID- 12080066 TI - The processing of Holliday junctions by BLM and WRN helicases is regulated by p53. AB - BLM, WRN, and p53 are involved in the homologous DNA recombination pathway. The DNA structure-specific helicases, BLM and WRN, unwind Holliday junctions (HJ), an activity that could suppress inappropriate homologous recombination during DNA replication. Here, we show that purified, recombinant p53 binds to BLM and WRN helicases and attenuates their ability to unwind synthetic HJ in vitro. The p53 248W mutant reduces abilities of both to bind HJ and inhibit helicase activities, whereas the p53 273H mutant loses these abilities. Moreover, full-length p53 and a C-terminal polypeptide (residues 373-383) inhibit the BLM and WRN helicase activities, but phosphorylation at Ser(376) or Ser(378) completely abolishes this inhibition. Following blockage of DNA replication, Ser(15) phospho-p53, BLM, and RAD51 colocalize in nuclear foci at sites likely to contain DNA replication intermediates in cells. Our results are consistent with a novel mechanism for p53 mediated regulation of DNA recombinational repair that involves p53 post translational modifications and functional protein-protein interactions with BLM and WRN DNA helicases. PMID- 12080067 TI - Purification and characterization of a receptor for human parathyroid hormone and parathyroid hormone-related peptide. AB - The human parathyroid hormone (PTH) receptor (hPTH1R), containing a 9-amino acid sequence of rhodopsin at its C terminus, was transiently expressed in COS-7 cells and solubilized with 0.25% n-dodecyl maltoside. Approximately 18 microg of hPTH1R were purified to homogeneity per mg of crude membranes by single-step affinity chromatography using 1D4, a monoclonal antibody to a rhodopsin epitope. The N terminus of the hPTH1R is Tyr(23), consistent with removal of the 22-amino acid signal peptide. Comparisons of hPTH1R by quantitative immunoblotting and Scatchard analysis revealed that 75% of the receptors in membrane preparations were functional; there was little, if any, loss of functional receptors during purification. The binding affinity of the purified hPTH1R was slightly lower than membrane-embedded hPTH1R (K(d) = 16.5 +/- 1.3 versus 11.9 +/- 1.9 nm), and the purified receptors bound rat [Nle(8,21),Tyr(34)]PTH-(1-34)-NH(2) (PTH-(1-34)), and rat [Ile(5),Trp(23),Tyr(36)]PTHrP-(5-36)-NH(2) with indistinguishable affinity. Maximal displacement of (125)I-PTH-(1-34) binding by rat [alpha aminoisobutyric acid (Aib)(1,3),Nle(8),Gln(10),Har(11),Ala(12),Trp(14),Arg(19),Tyr(21)]PTH-(1-21) NH(2) and rat [Aib(1,3),Gln(10),Har(11),Ala(12),Trp(14)]PTH-(1-14)-NH(2) of 80 and 10%, respectively, indicates that both N-terminal and juxtamembrane ligand binding determinants are functional in the purified hPTH1R. Finally, PTH stimulated [(35)S]GTP gamma S incorporation into G alpha(s) in a time- and dose dependent manner, when recombinant hPTH1R, G alpha(s)-, and beta gamma-subunits were reconstituted in phospholipid vesicles. The methods described will enable structural studies of the hPTH1R, and they provide an efficient and general technique to purify proteins, particularly those of the class II G protein coupled receptor family. PMID- 12080068 TI - Distinct interactions of GTP, UTP, and CTP with G(s) proteins. AB - Early studies showed that in addition to GTP, the pyrimidine nucleotides UTP and CTP support activation of the adenylyl cyclase (AC)-stimulating G(s) protein. The aim of this study was to elucidate the mechanism by which UTP and CTP support G(s) activation. As models, we used S49 wild-type lymphoma cells, representing a physiologically relevant system in which the beta(2)-adrenoreceptor (beta(2)AR) couples to G(s), and Sf9 insect cell membranes expressing beta(2)AR-Galpha(s) fusion proteins. Fusion proteins provide a higher sensitivity for the analysis of beta(2)AR-G(s) coupling than native systems. Nucleoside 5'-triphosphates (NTPs) supported agonist-stimulated AC activity in the two systems and basal AC activity in membranes from cholera toxin-treated S49 cells in the order of efficacy GTP > or = UTP > CTP > ATP (ineffective). NTPs disrupted high affinity agonist binding in beta(2)AR-Galpha(s) in the order of efficacy GTP > UTP > CTP > ATP (ineffective). In contrast, the order of efficacy of NTPs as substrates for nucleoside diphosphokinase, catalyzing the formation of GTP from GDP and NTP was ATP > or = UTP > or = CTP > or = GTP. NTPs inhibited beta(2)AR-Galpha(s) catalyzed [gamma-(32)P]GTP hydrolysis in the order of potency GTP > UTP > CTP. Molecular dynamics simulations revealed that UTP is accommodated more easily within the binding pocket of Galpha(s) than CTP. Collectively, our data indicate that GTP, UTP, and CTP interact differentially with G(s) proteins and that transphosphorylation of GDP to GTP is not involved in this G protein activation. In certain cell systems, intracellular UTP and CTP concentrations reach approximately 10 nmol/mg of protein and are higher than intracellular GTP concentrations, indicating that G protein activation by UTP and CTP can occur physiologically. G protein activation by UTP and CTP could be of particular importance in pathological conditions such as cholera and Lesch-Nyhan syndrome. PMID- 12080069 TI - Glia- and neuron-specific expression of the renin-angiotensin system in brain alters blood pressure, water intake, and salt preference. AB - The purpose of this study is to examine the regulation of blood pressure and fluid and electrolyte homeostasis in mice overexpressing angiotensin II (Ang-II) in the brain and to determine whether there are significant physiologic differences in Ang-II production in neurons or glia. Therefore, we generated and characterized transgenic mice overexpressing human renin (hREN) under the control of the glial fibrillary acidic protein (GFAP) promoter (GFAP-hREN) and synapsin-I promoter (SYN-hREN) and bred them with mice expressing human angiotensinogen (hAGT) under the control of the same promoters (GFAP-hAGT and SYN-hAGT). Both GFAP-hREN and SYN-hREN mice exhibited the highest hREN mRNA expression in the brain and had undetectable levels of hREN protein in the systemic circulation. In the brain of GFAP-hREN and SYN-hREN mice, hREN protein was observed almost exclusively in astrocytes and neurons, respectively. Transgenic mice overexpressing both hREN and hAGT transgenes in either glia or neurons were moderately hypertensive. In the glia-targeted mice, blood pressure could be corrected by intracerebroventricular injection of the Ang-II type 1 receptor antagonist losartan, and intravenous injection of a ganglion blocking agent, but not an arginine vasopressin V1 receptor antagonist, lowered blood pressure. These data suggest that stimulation of Ang-II type 1 receptors in the brain by Ang-II derived from local synthesis of renin and angiotensinogen can cause an elevation in blood pressure via a mechanism involving enhanced sympathetic outflow. Glia- and neuron-targeted mice also exhibited an increase in drinking volume and salt preference, suggesting that chronic overexpression of renin and angiotensinogen locally in the brain can result in hypertension and alterations in fluid homeostasis. PMID- 12080070 TI - The turn motif is a phosphorylation switch that regulates the binding of Hsp70 to protein kinase C. AB - Heat shock proteins play central roles in ensuring the correct folding and maturation of cellular proteins. Here we show that the heat shock protein Hsp70 has a novel role in prolonging the lifetime of activated protein kinase C. We identified Hsp70 in a screen for binding partners for the carboxyl terminus of protein kinase C. Co-immunoprecipitation experiments revealed that Hsp70 specifically binds the unphosphorylated turn motif (Thr(641) in protein kinase C beta II), one of three priming sites phosphorylated during the maturation of protein kinase C family members. The interaction of Hsp70 with protein kinase C can be abolished in vivo by co-expression of fusion proteins encoding the carboxyl terminus of protein kinase C or the carboxyl terminus of Hsp70. Pulse chase experiments reveal that Hsp70 does not regulate the maturation of protein kinase C: the rate of processing by phosphorylation is the same in the presence or absence of disrupting constructs. Rather, Hsp70 prolongs the lifetime of mature protein kinase C; disruption of the interaction promotes the accumulation of matured and then dephosphorylated protein kinase C in the detergent-insoluble fraction of cells. Furthermore, studies with K562 cells reveal that disruption of the interaction with Hsp70 slows the protein kinase C beta II-mediated recovery of cells from PMA-induced growth arrest. Last, we show that other members of the AGC superfamily (Akt/protein kinase B and protein kinase A) also bind Hsp70 via their unphosphorylated turn motifs. Our data are consistent with a model in which Hsp70 binds the dephosphorylated carboxyl terminus of mature protein kinase C, thus stabilizing the protein and allowing re-phosphorylation of the enzyme. Disruption of this interaction prevents re-phosphorylation and targets the enzyme for down-regulation. PMID- 12080071 TI - Tat HIV-1 primary and tertiary structures critical to immune response against non homologous variants. AB - Clinical studies show that in the absence of anti-retroviral therapy an immune response against the human immunodeficiency virus type 1 (HIV-1), transacting transcriptional activator (Tat) protein correlates with long term non progression. The purpose of this study is to try to understand what can trigger an effective immune response against Tat. We used five Tat variants from HIV strains identified in different parts of the world and showed that mutations of as much as 38% exist without any change in activity. Rabbit sera were raised against Tat variants identified in rapid-progressor patients (Tat HXB2, a European variant and Tat Eli, an African variant) and a long term non-progressor patient (Tat Oyi, an inactive African variant). Enzyme-linked immunosorbent assay (ELISA) results showed that anti-Tat Oyi serum had the highest antibody titer and was the only one to have a broad antibody response against heterologous Tat variants. Surprisingly, Tat HXB2 was better recognized by anti-Tat Oyi serum compared with anti-Tat HXB2 serum. Western blots showed that non-homologous Tat variants were recognized by antibodies directed against conformational epitopes. This study suggests that the primary and tertiary structures of the Tat variant from the long term non-progressor patient are critical to the induction of a broad and effective antibody response against Tat. PMID- 12080073 TI - Stationary and time-resolved resonance Raman spectra of His77 and Met95 mutants of the isolated heme domain of a direct oxygen sensor from Escherichia coli. AB - The heme environments of Met(95) and His(77) mutants of the isolated heme-bound PAS domain (Escherichia coli DOS PAS) of a direct oxygen sensing protein from E. coli (E. coli DOS) were investigated with resonance Raman (RR) spectroscopy and compared with the wild type (WT) enzyme. The RR spectra of both the reduced and oxidized WT enzyme were characteristic of six-coordinate low spin heme complexes from pH 4 to 10. The time-resolved RR spectra of the photodissociated CO-WT complex had an iron-His stretching band (nu(Fe-His)) at 214 cm(-1), and the nu(Fe CO) versus nu(CO) plot of CO-WT E. coli DOS PAS fell on the line of His coordinated heme proteins. The photodissociated CO-H77A mutant complex did not yield the nu(Fe-His) band but gave a nu(Fe-Im) band in the presence of imidazole. The RR spectrum of the oxidized M95A mutant was that of a six-coordinate low spin complex (i.e. the same as that of the WT enzyme), whereas the reduced mutant appeared to contain a five-coordinate heme complex. Taken together, we suggest that the heme of the reduced WT enzyme is coordinated by His(77) and Met(95), and that Met(95) is displaced by CO and O(2). Presumably, the protein conformational change that occurs upon exchange of an unknown ligand for Met(95) following heme reduction may lead to activation of the phosphodiesterase domain of E. coli DOS. PMID- 12080072 TI - Mitochondria are direct targets of the lipoxygenase inhibitor MK886. A strategy for cell killing by combined treatment with MK886 and cyclooxygenase inhibitors. AB - We have investigated the mitochondrial and cellular effects of the lipoxygenase inhibitor MK886. Low concentrations (1 microM) of MK886 selectively sensitized the permeability transition pore (PTP) to opening, whereas higher concentrations of MK886 (10 microM) caused depolarization through combination of an ionophoretic effect with inhibition of respiration. MK886 killed prostate cancer PC3 cells only at the higher, toxic concentration (10 microM), whereas the lower concentration (1 microM) had no major effect on cell survival. However, 1 microM MK886 alone demonstrably induced PTP-dependent mitochondrial dysfunction; and it caused cell death through the mitochondrial pathway when it was used in combination with the cyclooxygenase inhibitor, indomethacin, which had no effects per se. Treatment with 1 microM MK886 plus indomethacin sensitized cells to killing by exogenous arachidonic acid, which induces PTP opening and cytochrome c release (Scorrano, L., Penzo, D., Petronilli, V., Pagano, F., and Bernardi, P. (2001) J. Biol. Chem. 276, 12035-12040). Combination of MK886 and cyclooxygenase inhibitors may represent a viable therapeutic strategy to force cell death through the mitochondrial pathway. This approach should be specifically useful to kill cells possessing a high flux of arachidonic acid and its metabolites like prostate and colon cancer cells. PMID- 12080074 TI - The Srk1 protein kinase is a target for the Sty1 stress-activated MAPK in fission yeast. AB - The fission yeast stress-activated Sty1/Spc1 MAPK pathway responds to a similar range of stresses as do the mammalian p38 and SAPK/JNK MAPK pathways. In addition, sty1(-) cells are sterile and exhibit a G(2) cell cycle delay, indicating additional roles of Sty1 in meiosis and cell cycle progression. To identify novel proteins involved in stress responses, a microarray analysis of the Schizosaccharomyces pombe genome was performed to find genes that are up regulated following exposure to stress in a Sty1-dependent manner. One such gene identified, srk1(+) (Sty1-regulated kinase 1), encodes a putative serine/threonine kinase homologous to mammalian calmodulin kinases. At the C terminus of Srk1 is a putative MAPK binding motif similar to that in the p38 substrates, MAPK-activated protein kinases 2 and 3. Indeed, we find that Srk1 is present in a complex with the Sty1 MAPK and is directly phosphorylated by Sty1. Furthermore, upon stress, Srk1 translocates from the cytoplasm to the nucleus in a process that is dependent on the Sty1 MAPK. Finally, we show that Srk1 has a role in regulating meiosis in fission yeast; following nitrogen limitation, srk1( ) cells enter meiosis significantly faster than wild-type cells and overexpression of srk1(+) inhibits the nitrogen starvation-induced arrest in G(1). PMID- 12080075 TI - Concurrent translocation of multiple polypeptide chains through the proteasomal degradation channel. AB - The proteasome can actively unfold proteins by sequentially unraveling their substrates from the attachment point of the degradation signal. To investigate the steric constraints imposed on substrate proteins during their degradation by the proteasome, we constructed a model protein in which specific parts of the polypeptide chain were covalently connected through disulfide bridges. The cross linked model proteins were fully degraded by the proteasome, but two or more cross-links retarded the degradation slightly. These results suggest that the pore of the proteasome allows the concurrent passage of at least three stretches of a polypeptide chain. A degradation channel that can tolerate some steric bulk may reconcile the two opposing needs for degradation that is compartmentalized to avoid aberrant proteolysis yet able to handle a range of substrates of various sizes. PMID- 12080076 TI - Molecular cloning and characterization of chondroitin-4-O-sulfotransferase-3. A novel member of the HNK-1 family of sulfotransferases. AB - We have identified and characterized an N-acetylgalactosamine-4-O sulfotransferase designated chondroitin-4-sulfotransferase-3 (C4ST-3) (GenBank accession number AY120869) based on its homology to HNK-1 sulfotransferase (HNK-1 ST). The cDNA predicts an open reading frame encoding a type II membrane protein of 341 amino acids with a 12-amino acid cytoplasmic domain and a 311-amino acid luminal domain containing a single potential N-linked glycosylation site. C4ST-3 has the greatest amino acid sequence identity when aligned with chondroitin-4-O sulfotransferase 1 (C4ST-1) (45%) but also shows significant amino acid identity with chondroitin-4-O-sulfotransferase 2 (C4ST-2) (27%), dermatan-4-O sulfotransferase 1 (29%), HNK-1 ST (26%), N-acetylgalactosamine-4-O sulfotransferase 1 (26%), and N-acetylgalactosamine-4-O-sulfotransferase 2 (23%). C4ST-3 transfers sulfate to the C-4 hydroxyl of beta1,4-linked GalNAc that is substituted with a beta-linked glucuronic acid at the C-3 hydroxyl. The open reading frame of C4ST-3 is encoded by three exons located on human chromosome 3q21.3. Northern blot analysis reveals a single 2.1-kilobase transcript. C4ST-3 message is expressed in adult liver and at lower levels in adult kidney, lymph nodes, and fetal liver. Although C4ST-3 and C4ST-1 have similar specificities, the highly restricted pattern of expression seen for C4ST-3 suggests that it has a different role than C4ST-1. PMID- 12080077 TI - Novel protein kinase C isoforms regulate human keratinocyte differentiation by activating a p38 delta mitogen-activated protein kinase cascade that targets CCAAT/enhancer-binding protein alpha. AB - The novel protein kinase C (nPKC) isoforms are important regulators of human involucrin (hINV) gene expression during keratinocyte differentiation (Efimova, T., and Eckert, R. L. (2000) J. Biol. Chem. 275, 1601-1607). Although the regulatory mechanism involves mitogen-activated protein kinase (MAPK) activation, the role of individual MAPK isoforms has not been elucidated. We therefore examined the effects of individual nPKCs on MAPK activation. We observe unique changes whereby nPKC expression simultaneously increases p38 activity and decreases ERK1 and ERK2 activity. Although p38 alpha, p38 beta, and p38 delta are expressed in keratinocytes, only a single isoform, p38 delta, accounts for the increased p38 activity. Parallel studies indicate that this isoform is also activated by treatment with the keratinocyte regulatory agents, 12-O tetradecanoylphorbol-13-acetate, calcium, and okadaic acid. These changes in MAPK activity are associated with increased C/EBP alpha transcription factor expression and DNA binding to the hINV promoter and increased hINV gene expression. Expression of PKC delta, PKC epsilon, or PKC eta causes a 10-fold increase in hINV promoter activity, whereas C/EBP alpha expression produces a 25 fold increase. However, simultaneous expression of both proteins causes a synergistic 100-fold increase in promoter activity. These responses are eliminated by the dominant-negative C/EBP isoform, GADD153, and are also inhibited by dominant-negative forms of Ras, MEKK1, MEK3, and p38. These results suggest that the nPKC isoforms produce a unique shift in MAPK activity via a Ras, MEKK1, MEK3 pathway, to increase p38 delta and inhibit ERK1/2 and ultimately increase C/EBP alpha binding to the hINV promoter and hINV gene expression. PMID- 12080078 TI - Phosphorylation of pyrimidine L-deoxynucleoside analog diphosphates. Kinetics of phosphorylation and dephosphorylation of nucleoside analog diphosphates and triphosphates by 3-phosphoglycerate kinase. AB - Anticancer and antiviral D- and L-nucleoside analogs are phosphorylated stepwise in the cells to the pharmacologically active triphosphate metabolites. We recently reported that in the last step, L-deoxynucleoside analog diphosphates are phosphorylated by 3-phosphoglycerate kinase (PGK). To explain the preference of PGK for L- over D-deoxynucleoside analog diphosphates, the kinetics of their phosphorylation were compared with the dephosphorylation of the respective triphosphates using recombinant human PGK. The results attributed favorable phosphorylation of L-deoxynucleoside analog diphosphates by PGK to differences in k(cat), which were consequences of varied orientations of the sugar and diphosphates in the catalytic site of PGK. The amino acids involved in the catalytic reaction of PGK (including Glu(344), Lys(220), and Asn(337)) were therefore mutated. The impact of mutations on the phosphorylation of L- and D deoxynucleoside analog diphosphates was different from those on dephosphorylation of the respective triphosphates. This suggested that the interactions of the nucleoside analogs with amino acids during the transition state are different in the phosphorylation and dephosphorylation reactions. Thus, reversible action of the enzyme may not involve the same configuration of the active site. Furthermore, the amino acid determinants of the action of PGK for L deoxynucleotides were not the same as for the D-deoxynucleotides. This study also suggests the potential impact of nucleoside analog diphosphates and triphosphates on the multiple cellular functions of PGK, which may contribute to the action of the analogs. PMID- 12080079 TI - Selective inhibition of dipeptidyl peptidase I, not caspases, prevents the partial processing of procaspase-3 in CD3-activated human CD8(+) T lymphocytes. AB - Activation of primary human T cells by anti-CD3 and interleukin-2 resulted in partial processing of procaspase-3 in activated nonapoptotic (Delta Psi(m)high) CD8(+) T cells but not in CD4(+) T cells. Apical caspases-8 and -9 were not activated, and Bid was not processed to truncated Bid. Boc-D.fmk, a broad spectrum caspase inhibitor, did not prevent this process, whereas GF.dmk, a selective inhibitor of dipeptidyl peptidase I, was effective. Dipeptidyl peptidase I is required for the activation of granule-associated serine proteases. It is enriched in the cytolytic granules of cytotoxic lymphocytes, where it promotes the proteolytic activation of progranzymes A and B. Inhibition of granzyme B (GrB)-like serine proteases by Z-AAD.cmk prevented partial processing of procapase-3, whereas inhibition of GrA activity by D-FPR.cmk had no effect. Specific inhibitors of other lysosomal proteases such as cathepsins B, L, and D did not interfere in this event. Patients with Chediak-Higashi syndrome or with perforin deficiency also displayed partial processing of procaspase-3, excluding the involvement of granule exocytosis for the delivery of the serine protease in cause. The p20/p12 processing pattern of procaspase-3 in our model points to GrB, the sole serine protease with caspase activity. Small amounts of GrB were indeed exported from cytolytic granules to the cytosol of a significant fraction of GrB-positive cells. PMID- 12080080 TI - Monensin mediates a rapid and selective transport of Pb(2+). Possible application of monensin for the treatment of Pb(2+) intoxication. AB - The carboxylic acid ionophore monensin, known as an electroneutral Na(+) ionophore, an anticoccidial agent, and a growth-promoting feed additive in agriculture, is shown to be highly efficient as an ionophore for Pb(2+) and to be highly selective for Pb(2+) compared with other divalent cations. Monensin transports Pb(2+) by an electroneutral mechanism in which the complex PbMonOH is the transporting species. Electrogenic transport via the species PbMon(+) may also be possible. Monensin catalyzed Pb(2+) transport is little affected by Ca(2+), Mg(2+), or K(+) concentrations that are encountered in living systems. Na(+) is inhibitory, but its effectiveness at 100 mm does not exceed approximately 50%. The poor activity of monensin as an ionophore for divalent cations other than Pb(2+) is consistent with the pattern of complex formation constants observed in the mixed solvent 80% methanol/water. This pattern also explains why Ca(2+), Mg(2+), and K(+) are ineffective as inhibitors of Pb(2+) transport, but it does not fully explain the actions of Na(+), where kinetic features of the transport mechanism may also be important. When given to rats at 100 ppm in feed together with Pb(2+) at 100 ppm in drinking water, monensin reduces Pb accumulation in several organs and tissues. It also accelerates the excretion of Pb that was accumulated previously and produces this effect without depleting the organs of zinc or copper. Monensin, used alone or in combination with other agents, may be useful for the treatment of Pb intoxication. PMID- 12080081 TI - Epithelial inducible nitric-oxide synthase is an apical EBP50-binding protein that directs vectorial nitric oxide output. AB - Nitric oxide (NO), produced via inducible NO synthase (iNOS), can modulate polarized epithelial processes such as solute transport. Given the high reactivity of NO, we hypothesized that optimal NO regulation of polarized epithelial functions is achieved through compartmentalization of iNOS, allowing local NO delivery to its molecular targets. Here, we show that iNOS localizes to the apical domain of epithelial cells within a submembranous protein complex tightly bound to cortical actin. We further show that iNOS can bind to the apical PDZ protein, EBP50 (ezrin-radixin-moesin-binding phosphoprotein 50), an interaction that is dependent on the last three COOH-terminal amino acids of iNOS, SAL, but requires the presence of additional unknown cellular proteins. Mutation of these three COOH-terminal residues abolishes the iNOS-EBP50 interaction and disrupts the apical association of iNOS in transfected cells, showing that this COOH-terminal motif is essential for the correct localization of iNOS in epithelial cells. Apically localized iNOS directs vectorial NO production at the apical proximal tubule epithelial cell surface. These studies define human epithelial iNOS as an apical EBP50-binding protein and suggest that the physical association of iNOS with EBP50 might allow precise NO modulation of EBP50-associated protein functions. PMID- 12080082 TI - Differential sensitivity of guanylyl cyclase and mitochondrial respiration to nitric oxide measured using clamped concentrations. AB - Nitric oxide (NO) signal transduction may involve at least two targets: the guanylyl cyclase-coupled NO receptor (NO(GC)R), which catalyzes cGMP formation, and cytochrome c oxidase, which is responsible for mitochondrial O(2) consumption and which is inhibited by NO in competition with O(2). Current evidence indicates that the two targets may be similarly sensitive to NO, but quantitative comparison has been difficult because of an inability to administer NO in known, constant concentrations. We addressed this deficiency and found that purified NO(GC)R was about 100-fold more sensitive to NO than reported previously, 50% of maximal activity requiring only 4 nm NO. Conversely, at physiological O(2) concentrations (20-30 microM), mitochondrial respiration was 2-10-fold less sensitive to NO than estimated beforehand. The two concentration-response curves showed minimal overlap. Accordingly, an NO concentration maximally active on the NO(GC)R (20 nm) inhibited respiration only when the O(2) concentration was pathologically low (50% inhibition at 5 microM O(2)). Studies on brain slices under conditions of maximal stimulation of endogenous NO synthesis suggested that the local NO concentration did not rise above 4 nm. It is concluded that under physiological conditions, at least in brain, NO is constrained to target the NO(GC)R without inhibiting mitochondrial respiration. PMID- 12080083 TI - How oxygen makes its presence felt. PMID- 12080084 TI - FGF signaling pathways in endochondral and intramembranous bone development and human genetic disease. PMID- 12080085 TI - FIH-1 is an asparaginyl hydroxylase enzyme that regulates the transcriptional activity of hypoxia-inducible factor. AB - Mammalian cells adapt to hypoxic conditions through a transcriptional response pathway mediated by the hypoxia-inducible factor, HIF. HIF transcriptional activity is suppressed under normoxic conditions by hydroxylation of an asparagine residue within its C-terminal transactivation domain, blocking association with coactivators. Here we show that the protein FIH-1, previously shown to interact with HIF, is an asparaginyl hydroxylase. Like known hydroxylase enzymes, FIH-1 is an Fe(II)-dependent enzyme that uses molecular O(2) to modify its substrate. Together with the recently discovered prolyl hydroxylases that regulate HIF stability, this class of oxygen-dependent enzymes comprises critical regulatory components of the hypoxic response pathway. PMID- 12080086 TI - Mammalian cell size is controlled by mTOR and its downstream targets S6K1 and 4EBP1/eIF4E. AB - The coordinated action of cell cycle progression and cell growth (an increase in cell size and cell mass) is critical for sustained cellular proliferation, yet the biochemical signals that control cell growth are poorly defined, particularly in mammalian systems. We find that cell growth and cell cycle progression are separable processes in mammalian cells and that growth to appropriate cell size requires mTOR- and PI3K-dependent signals. Expression of a rapamycin-resistant mutant of mTOR rescues the reduced cell size phenotype induced by rapamycin in a kinase-dependent manner, showing the evolutionarily conserved role of mTOR in control of cell growth. Expression of S6K1 mutants that possess partial rapamycin resistant activity or overexpression of eIF4E individually and additively partially rescues the rapamycin-induced decrease in cell size. In the absence of rapamycin, overexpression of S6K1 or eIF4E increases cell size, and, when coexpressed, they cooperate to increase cell size further. Expression of a phosphorylation site-defective mutant of 4EBP1 that constitutively binds the eIF4E-Cap complex to inhibit translation initiation reduces cell size and blocks eIF4E effects on cell size. These data show that mTOR signals downstream to at least two independent targets, S6K1 and 4EBP1/eIF4E, that function in translational control to regulate mammalian cell size. PMID- 12080087 TI - Neurogenin 3 is essential for the proper specification of gastric enteroendocrine cells and the maintenance of gastric epithelial cell identity. AB - The notch signaling pathway is essential for the endocrine cell fate in various tissues including the enteroendocrine system of the gastrointestinal tract. Enteroendocrine cells are one of the four major cell types found in the gastric epithelium of the glandular stomach. To understand the molecular basis of enteroendocrine cell development, we have used gene targeting in mouse embryonic stem cells to derive an EGFP-marked null allele of the bHLH transcription factor, neurogenin 3 (ngn3). In ngn3(-/-) mice, glucagon secreting A-cells, somatostatin secreting D-cells, and gastrin secreting G-cells are absent from the epithelium of the glandular stomach, whereas the number of serotonin-expressing enterochromaffin (EC) cells is decreased dramatically. In addition, ngn3(-/-) mice display intestinal metaplasia of the gastric epithelium. Thus, ngn3 is required for the differentiation of enteroendocrine cells in the stomach and the maintenance of gastric epithelial cell identity. PMID- 12080088 TI - Characterization of HCP-6, a C. elegans protein required to prevent chromosome twisting and merotelic attachment. AB - Previous studies of mitosis show that capture of single kinetochores by microtubules from both centrosomes (merotelic orientation) is a major cause of aneuploidy. We have characterized hcp-6, a temperature-sensitive chromosome segregation mutant in C. elegans that exhibits chromosomes attached to both poles via a single sister kinetochore. We demonstrate that the primary defect in this mutant is a failure to fully condense chromosomes during prophase. Although centromere formation and sister centromere resolution remain unaffected in hcp-6, the chromosomes lack the rigidity of wild-type chromosomes and twist around the long axis of the chromosome. As such, they are unable to establish a proper orientation at prometaphase, allowing individual kinetochores to be captured by microtubules from both poles. We therefore propose that chromosome rigidity plays an essential role in maintaining chromosome orientation to prevent merotelic capture. PMID- 12080089 TI - JunB potentiates function of BRCA1 activation domain 1 (AD1) through a coiled coil-mediated interaction. AB - BRCA1 is involved in the regulation of multiple nuclear events including transcription. AD1, one of the two trans-activation domains in BRCA1, stimulates transcription in a cell context-dependent manner. Here, it is shown that BRCA1 interacts with Jun proteins via a coiled-coil motif in AD1 and the basic leucine zipper (bZIP) region of the Jun proteins. The Jun-interacting domain in BRCA1 is critical for AD1-mediated transcriptional activation. In particular, the strength of AD1 in transcriptional activation is limited by the JunB level and ectopic expression of JunB potentiates the transcriptional activity of AD1. Furthermore, JunB mRNA expression is down-regulated in many ovarian tumor tissues examined. Thus, the coiled-coil-mediated cooperation between BRCA1 and JunB may facilitate the function of these proteins in tissue-specific transcriptional regulation and tumor suppression. PMID- 12080090 TI - Lysine methylation within the globular domain of histone H3 by Dot1 is important for telomeric silencing and Sir protein association. AB - The amino-terminal histone tails are subject to covalent post-translational modifications such as acetylation, methylation, and phosphorylation. In the histone code hypothesis, these exposed and unstructured histone tails are accessible to a repertoire of regulatory factors that specifically recognize the various modified histones, thereby generating altered chromatin structures that mediate specific biological responses. Here, we report that lysine (Lys) 79 of histone H3, which resides in the globular domain, is methylated in eukaryotic organisms. In the yeast Saccharomyces cerevisiae, Lys 79 of histone H3 is methylated by Dot1, a protein shown previously to play a role in telomeric silencing. Mutations of Lys 79 of histone H3 and mutations that abolish the catalytic activity of Dot1 impair telomeric silencing, suggesting that Dot1 mediates telomeric silencing largely through methylation of Lys 79. This defect in telomeric silencing might reflect an interaction between Sir proteins and Lys 79, because dot1 and Lys 79 mutations weaken the interaction of Sir2 and Sir3 with the telomeric region in vivo. Our results indicate that histone modifications in the core globular domain have important biological functions. PMID- 12080091 TI - Rap1-Sir4 binding independent of other Sir, yKu, or histone interactions initiates the assembly of telomeric heterochromatin in yeast. AB - In Saccharomyces cerevisiae, heterochromatin-like regions are found near telomeres and at the silent mating-type loci, where they can repress genes in an epigenetic manner. Several proteins are involved in telomeric heterochromatin structure including Rap1, Sir2, Sir3, Sir4, yKu70 (Hdf1), yKu80 (Hdf2), and the N termini of histones H3 and H4. By recognizing cis-acting DNA-binding sites, Rap1 is believed to recruit Sir and other silencing proteins and determine where heterochromatin forms. The integrity of heterochromatin also requires the binding of Sir proteins to histones that may form a scaffold for Sir protein interactions with chromatin. In this study we describe how the heterochromatin complex may form initially and how it differs from the complex that spreads along the chromosome. We found that close to the telomere end, Sir4 can bind Rap1 independently of Sir2, Sir3, yKu70/yKu80, and the intact H4 N terminus. In contrast, Sir4 binding requires all of the silencing factors further along telomeric heterochromatin. These data indicate that Sir4 binding to Rap1 initiates the sequential association of Sir and other proteins, allowing the subsequent spreading of the heterochromatin proteins along the chromosome. PMID- 12080092 TI - The barrier function of an insulator couples high histone acetylation levels with specific protection of promoter DNA from methylation. AB - Stably integrated transgenes flanked by the chicken beta-globin HS4 insulator are protected against chromosomal position effects and gradual extinction of expression during long-term propagation in culture. To investigate the mechanism of action of this insulator, we used bisulfite genomic sequencing to examine the methylation of individual CpG sites within insulated transgenes, and compared this with patterns of histone acetylation. Surprisingly, although the histones of the entire insulated transgene are highly acetylated, only a specific region in the promoter, containing binding sites for erythroid-specific transcription factors, is highly protected from DNA methylation. This critical region is methylated in noninsulated and inactive lines. MBD3 and Mi-2, subunits of the Mi 2/NuRD repressor complex, are bound in vivo to these silenced noninsulated transgenes. In contrast, insulated cell lines do not show any enrichment of Mi 2/NuRD proteins very late in culture. In addition to the high levels of histone acetylation observed across the entire insulated transgene, significant peaks of H3 acetylation are present over the HS4 insulator elements. Targeted histone acetylation by the chicken beta-globin insulator occurs independently of gene transcription and does not require the presence of a functional enhancer. We suggest that this acetylation is in turn responsible for the maintenance of a region of unmethylated DNA over the promoter. Whereas DNA methylation often leads to histone deacetylation, here acetylation appears to prevent methylation. PMID- 12080093 TI - In the complex family of heat stress transcription factors, HsfA1 has a unique role as master regulator of thermotolerance in tomato. AB - We generated transgenic tomato plants with altered expression of heat stress transcription factor HsfA1. Plants with 10-fold overexpression of HsfA1 (OE plants) were characterized by a single HsfA1 transgene cassette, whereas plants harboring a tandem inverted repeat of the cassette showed cosuppression (CS plants) by posttranscriptional silencing of the HsfA1 gene connected with formation of small interfering RNAs. Under normal growth conditions, major developmental parameters were similar for wild-type (WT), OE, and CS plants. However, CS plants and fruits were extremely sensitive to elevated temperatures, because heat stress-induced synthesis of chaperones and Hsfs was strongly reduced or lacking. Despite the complexity of the plant Hsf family with at least 17 members in tomato, HsfA1 has a unique function as master regulator for induced thermotolerance. Using transient reporter assays with mesophyll protoplasts from WT tomato, we demonstrated that plasmid-encoded HsfA1 and HsfA2 were well expressed. However, in CS protoplasts the cosuppression phenomenon was faithfully reproduced. Only transformation with HsfA2 expression plasmid led to normal expression of the transcription factor and reporter gene activation, whereas even high amounts of HsfA1 expression plasmids were silenced. Thermotolerance in CS protoplasts was restored by plasmid-borne HsfA2, resulting in expression of chaperones, thermoprotection of firefly luciferase, and assembly of heat stress granules. PMID- 12080095 TI - A mode of thought in excitation-contraction coupling. PMID- 12080096 TI - Unclogging a pipe: potassium channel pinball. PMID- 12080094 TI - Targeted mutagenesis by homologous recombination in D. melanogaster. AB - We used a recently developed method to produce mutant alleles of five endogenous Drosophila genes, including the homolog of the p53 tumor suppressor. Transgenic expression of the FLP site-specific recombinase and the I-SceI endonuclease generates extrachromosomal linear DNA molecules in vivo. These molecules undergo homologous recombination with the corresponding chromosomal locus to generate targeted alterations of the host genome. The results address several questions about the general utility of this technique. We show that genes not near telomeres can be efficiently targeted; that no knowledge of the mutant phenotype is needed for targeting; and that insertional mutations and allelic substitutions can be easily produced. PMID- 12080097 TI - Reduced transition between open and inactivated channel states underlies 5HT increased I(Na+) in rat nociceptors. AB - We previously demonstrated that activation of a 5HT(4) receptor coupled cAMP dependent signaling pathway increases tetrodotoxin-resistant Na(+) current (I(Na)) in a nociceptor-like subpopulation of rat dorsal root ganglion cells (type 2). In the present study we used electrophysiology experiments and computer modeling studies to explore the mechanism(s) underlying the increase of I(Na) by 5HT. In electrophysiological experiments with type 2 dorsal root ganglion cells, 5HT increased peak I(Na) and the activation and inactivation rate, without significantly affecting the voltage dependency of activation or availability. Studies on the voltage dependency of channel availability, time course of removal of inactivation, and inactivation of evoked Na(+) currents suggested that there are at least two inactivation states of the Na(+) channel, one (I(fast)) that is induced and retrieved faster than the other (I(slow)). Long (1 s), but not short (60 or 100 ms), inactivating conditioning pulses (CPs) suppressed the 5HT-induced increase in I(Na). Computer modeling studies suggest that 5HT increased I(Na) mainly by decreasing the transition rate (k(OI1)) from an open state to I(fast). Furthermore, 5HT increased I(Na) activation and inactivation rates mainly by increasing the transition rate from closed to open (k(C3O)) and from I(fast) to I(slow) (k(I1I2)), respectively. The antagonism of the 5HT-induced increase in I(Na) by 1-s inactivation CPs may be due an enhancement of transitions from I(fast) to I(slow), via the increase in k(I1I2). This may deplete the pool of channels residing in I(fast), reducing the frequency of reopenings from I(fast), which offsets the increase in I(Na) produced by the reduction in k(OI1). The above findings fit well with previous studies showing that activation of the cAMP/PKA cascade simultaneously increases voltage sensitive tetrodotoxin resistant Na(+) conductance and inactivation rate in nociceptors. The antagonism of the effects of 5HT by long inactivation CPs suggests that drugs designed to induce and/or stabilize the I(slow) state might be useful for reducing hyperalgesia produced by inflammatory mediators. PMID- 12080098 TI - Sequence-dependent motions of DNA: a normal mode analysis at the base-pair level. AB - Computer simulation of the dynamic structure of DNA can be carried out at various levels of resolution. Detailed high resolution information about the motions of DNA is typically collected for the atoms in a few turns of double helix. At low resolution, by contrast, the sequence-dependence features of DNA are usually neglected and molecules with thousands of base pairs are treated as ideal elastic rods. The present normal mode analysis of DNA in terms of six base-pair "step" parameters per chain residue addresses the dynamic structure of the double helix at intermediate resolution, i.e., the mesoscopic level of a few hundred base pairs. Sequence-dependent effects are incorporated into the calculations by taking advantage of "knowledge-based" harmonic energy functions deduced from the mean values and dispersion of the base-pair "step" parameters in high-resolution DNA crystal structures. Spatial arrangements sampled along the dominant low frequency modes have end-to-end distances comparable to those of exact polymer models which incorporate all possible chain configurations. The normal mode analysis accounts for the overall bending, i.e., persistence length, of the double helix and shows how known discrepancies in the measured twisting constants of long DNA molecules could originate in the deformability of neighboring base pair steps. The calculations also reveal how the natural coupling of local conformational variables affects the global motions of DNA. Successful correspondence of the computed stretching modulus with experimental data requires that the DNA base pairs be inclined with respect to the direction of stretching, with chain extension effected by low energy transverse motions that preserve the strong van der Waals' attractions of neighboring base-pair planes. The calculations further show how one can "engineer" the macroscopic properties of DNA in terms of dimer deformability so that polymers which are intrinsically straight in the equilibrium state exhibit the mesoscopic bending anisotropy essential to DNA curvature and loop formation. PMID- 12080099 TI - A simple, mechanistic model for directional instability during mitotic chromosome movements. AB - During mitosis, chromosomes become attached to microtubules that emanate from the two spindle poles. Thereafter, a chromosome moves along these microtubule "tracks" as it executes a series of movements that bring it to the spindle equator. After the onset of anaphase, the sister chromatids separate and move to opposite spindle poles. These movements are often characterized by "directional instability" (a series of runs with approximately constant speed, punctuated by sudden reversals in the direction of movement). To understand mitosis, it is critical to describe the physical mechanisms that underlie the coordination of the forces that drive directional instability. We propose a simple mechanistic model that describes the origin of the forces that move chromosomes and the coordination of these forces to produce directional instability. The model demonstrates that forces, speeds, and direction of motion associated with prometaphase through anaphase chromosome movements can be predicted from the molecular kinetics of interactions between dynamic microtubules and arrays of microtubule binding sites that are linked to the chromosome by compliant elements. PMID- 12080100 TI - Termination of cardiac Ca(2+) sparks: an investigative mathematical model of calcium-induced calcium release. AB - A Ca(2+) spark arises when a cluster of sarcoplasmic reticulum (SR) channels (ryanodine receptors or RyRs) opens to release calcium in a locally regenerative manner. Normally triggered by Ca(2+) influx across the sarcolemmal or transverse tubule membrane neighboring the cluster, the Ca(2+) spark has been shown to be the elementary Ca(2+) signaling event of excitation-contraction coupling in heart muscle. However, the question of how the Ca(2+) spark terminates remains a central, unresolved issue. Here we present a new model, "sticky cluster," of SR Ca(2+) release that simulates Ca(2+) spark behavior and enables robust Ca(2+) spark termination. Two newly documented features of RyR behavior have been incorporated in this otherwise simple model: "coupled gating" and an opening rate that depends on SR lumenal [Ca(2+)]. Using a Monte Carlo method, local Ca(2+) induced Ca(2+) release from clusters containing between 10 and 100 RyRs is modeled. After release is triggered, Ca(2+) flux from RyRs diffuses into the cytosol and binds to intracellular buffers and the fluorescent Ca(2+) indicator fluo-3 to produce the model Ca(2+) spark. Ca(2+) sparks generated by the sticky cluster model resemble those observed experimentally, and Ca(2+) spark duration and amplitude are largely insensitive to the number of RyRs in a cluster. As expected from heart cell investigation, the spontaneous Ca(2+) spark rate in the model increases with elevated cytosolic or SR lumenal [Ca(2+)]. Furthermore, reduction of RyR coupling leads to prolonged model Ca(2+) sparks just as treatment with FK506 lengthens Ca(2+) sparks in heart cells. This new model of Ca(2+) spark behavior provides a "proof of principle" test of a new hypothesis for Ca(2+) spark termination and reproduces critical features of Ca(2+) sparks observed experimentally. PMID- 12080101 TI - Energy balance for analysis of complex metabolic networks. AB - Predicting behavior of large-scale biochemical networks represents one of the greatest challenges of bioinformatics and computational biology. Computational tools for predicting fluxes in biochemical networks are applied in the fields of integrated and systems biology, bioinformatics, and genomics, and to aid in drug discovery and identification of potential drug targets. Approaches, such as flux balance analysis (FBA), that account for the known stoichiometry of the reaction network while avoiding implementation of detailed reaction kinetics are promising tools for the analysis of large complex networks. Here we introduce energy balance analysis (EBA)--the theory and methodology for enforcing the laws of thermodynamics in such simulations--making the results more physically realistic and revealing greater insight into the regulatory and control mechanisms operating in complex large-scale systems. We show that EBA eliminates thermodynamically infeasible results associated with FBA. PMID- 12080102 TI - Stochastic properties of Ca(2+) release of inositol 1,4,5-trisphosphate receptor clusters. AB - Intracellular Ca(2+) release is controlled by inositol 1,4,5-trisphosphate (IP(3)) receptors or ryanodine receptors. These receptors are typically distributed in clusters with several or tens of channels. The random opening and closing of these channels introduces stochasticity into the elementary calcium release mechanism. Stochastic release events have been experimentally observed in a variety of cell types and have been termed sparks and puffs. We put forward a stochastic version of the Li-Rinzel model (the deactivation binding process is described by a Markovian scheme) and a computationally more efficient Langevin approach to model the stochastic Ca(2+) oscillation of single clusters. Statistical properties such as Ca(2+) puff amplitudes, lifetimes, and interpuff intervals are studied with both models and compared with experimental observations. For clusters with tens of channels, a simply decaying amplitude distribution is typically observed at low IP(3) concentration, while a single peak distribution appears at high IP(3) concentration. PMID- 12080103 TI - Orientation and conformation of a lipase at an interface studied by molecular dynamics simulations. AB - Electron density profiles calculated from molecular dynamics trajectories are used to deduce the orientation and conformation of Thermomyces lanuginosa lipase and a mutant adsorbed at an air-water interface. It is demonstrated that the profiles display distinct fine structures, which uniquely characterize enzyme orientation and conformation. The density profiles are, on the nanosecond timescale, determined by the average enzyme conformation. We outline a computational scheme that from a single molecular dynamics trajectory allows for extraction of electron density profiles referring to different orientations of the lipase relative to an implicit interface. Profiles calculated for the inactive and active conformations of the lipase are compared with experimental electron density profiles measured by x-ray reflectivity for the lipase adsorbed at an air-water interface. The experimental profiles contain less fine structural information than the calculated profiles because the resolution of the experiment is limited by the intrinsic surface roughness of water. Least squares fits of the calculated profiles to the experimental profiles provide areas per adsorbed enzyme and suggest that Thermomyces lanuginosa lipase adsorbs to the air-water interface in a semiopen conformation with the lid oriented away from the interface. PMID- 12080104 TI - Molecular dynamics study of the folding of hydrophobin SC3 at a hydrophilic/hydrophobic interface. AB - Hydrophobins are fungal proteins that self-assemble at hydrophilic/hydrophobic interfaces into amphipathic membranes. These assemblages are extremely stable and posses the remarkable ability to invert the polarity of the surface on which they are adsorbed. Neither the three-dimensional structure of a hydrophobin nor the mechanism by which they function is known. Nevertheless, there are experimental indications that the self-assembled form of the hydrophobins SC3 and EAS at a water/air interface is rich with beta-sheet secondary structure. In this paper we report results from molecular dynamics simulations, showing that fully extended SC3 undergoes fast (approximately 100 ns) folding at a water/hexane interface to an elongated planar structure with extensive beta-sheet secondary elements. Simulations in each of the bulk solvents result in a mainly unstructured globular protein. The dramatic enhancement in secondary structure, whether kinetic or thermodynamic in origin, highlights the role interfaces between phases with large differences in polarity can have on folding. The partitioning of the residue side chains to one of the two phases can serve as a strong driving force to initiate secondary structure formation. The interactions of the side-chains with the environment at an interface can also stabilize configurations that otherwise would not occur in a homogenous solution. PMID- 12080105 TI - Asymmetry of glia near central synapses favors presynaptically directed glutamate escape. AB - Recent findings demonstrate that synaptically released excitatory neurotransmitter glutamate activates receptors outside the immediate synaptic cleft and that the extent of such extrasynaptic actions is regulated by the high affinity glutamate uptake. The bulk of glutamate transporter systems are evenly distributed in the synaptic neuropil, and it is generally assumed that glutamate escaping the cleft affects pre- and postsynaptic receptors to a similar degree. To test whether this is indeed the case, we use quantitative electron microscopy and establish the stochastic pattern of glial occurrence in the three-dimensional (3D) vicinity of two common types of excitatory central synapses, stratum radiatum synapses in hippocampus and parallel fiber synapses in cerebellum. We find that the occurrence of glia postsynaptically is strikingly higher (3-4-fold) than presynaptically, in both types of synapses. To address the functional consequences of this asymmetry, we simulate diffusion and transport of synaptically released glutamate in these two brain areas using a detailed 3D compartmental model of the extracellular space with glutamate transporters arranged unevenly, in accordance with the obtained experimental data. The results predict that glutamate escaping the synaptic cleft is 2-4 times more likely to activate presynaptic compared to postsynaptic receptors. Simulations also show that postsynaptic neuronal transporters (EAAT4 type) at dendritic spines of cerebellar Purkinje cells exaggerate this asymmetry further. Our data suggest that the perisynaptic environment of these common central synapses favors fast presynaptic feedback in the information flow while preserving the specificity of the postsynaptic input. PMID- 12080106 TI - Membrane structure of the human immunodeficiency virus gp41 fusion domain by molecular dynamics simulation. AB - The structures of the 16-residue fusion domain (or fusion peptide, FP) of the human immunodeficiency virus gp41 fusion protein, two of its mutants, and a shortened peptide (5-16) were studied by molecular dynamics simulation in an explicit palmitoyloleoylphosphoethanolamine bilayer. The simulations showed that the active wild-type FP inserts into the bilayer approximately 44 degrees +/- 6 degrees with respect to the bilayer normal, whereas the inactive V2E and L9R mutants and the inactive 5 to 16 fragment lie on the bilayer surface. This is the first demonstration by explicit molecular dynamics of the oblique insertion of the fusion domain into lipid bilayers, and provides correlation between the mode of insertion and the fusogenic activity of these peptides. The membrane structure of the wild-type FP is remarkably similar to that of the influenza HA(2) FP as determined by nuclear magnetic resonance and electron spin resistance power saturation. The secondary structures of the wild-type FP and the two inactive mutants are quite similar, indicating that the secondary structure of this fusion domain plays little or no role in affecting the fusogenic activity of the fusion peptide. The insertion of the wild-type FP increases the thickness of the interfacial area of the bilayer by disrupting the hydrocarbon chains and extending the interfacial area toward the head group region, an effect that was not observed in the inactive FPs. PMID- 12080107 TI - Interaction of cardiotoxins with membranes: a molecular modeling study. AB - Incorporation of beta-sheet proteins into membrane is studied theoretically for the first time, and the results are validated by the direct experimental data. Using Monte Carlo simulations with implicit membrane, we explore spatial structure, energetics, polarity, and mode of insertion of two cardiotoxins with different membrane-destabilizing activity. Both proteins, classified as P- and S type cardiotoxins, are found to retain the overall "three-finger" fold interacting with membrane core and lipid/water interface by the tips of the "fingers" (loops). The insertion critically depends upon the structure, hydrophobicity, and electrostatics of certain regions. The simulations reveal apparently distinct binding modes for S- and P-type cardiotoxins via the first loop or through all three loops, respectively. This rationalizes an earlier empirical classification of cardiotoxins into S- and P-type, and provides a basis for the analysis of experimental data on their membrane affinities. Accomplished with our previous simulations of membrane alpha-helices, the computational method may be used to study partitioning of proteins with diverse folds into lipid bilayers. PMID- 12080109 TI - Simulations of molecular diffusion in lattices of cells: insights for NMR of red blood cells. AB - The pulsed field-gradient spin-echo (PGSE) nuclear magnetic resonance (NMR) experiment, conducted on a suspension of red blood cells (RBC) in a strong magnetic field yields a q-space plot consisting of a series of maxima and minima. This is mathematically analogous to a classical optical diffraction pattern. The method provides a noninvasive and novel means of characterizing cell suspensions that is sensitive to changes in cell shape and packing density. The positions of the features in a q-space plot characterize the rate of exchange across the membrane, cell dimensions, and packing density. A diffusion tensor, containing information regarding the diffusion anisotropy of the system, can also be derived from the PGSE NMR data. In this study, we carried out Monte Carlo simulations of diffusion in suspensions of "virtual" cells that had either biconcave disc (as in RBC) or oblate spheroid geometry. The simulations were performed in a PGSE NMR context thus enabling predictions of q-space and diffusion tensor data. The simulated data were compared with those from real PGSE NMR diffusion experiments on RBC suspensions that had a range of hematocrit values. Methods that facilitate the processing of q-space data were also developed. PMID- 12080108 TI - Pressure-induced water transport in membrane channels studied by molecular dynamics. AB - A method is proposed to measure the water permeability of membrane channels by means of molecular dynamics simulations. By applying a constant force to the bulk water molecules and a counter force on the complementary system, a hydrostatic pressure difference across the membrane can be established, producing a net directional water flow. The hydraulic or osmotic permeability can then be determined by the ratio of the water flux and the pressure difference. The method is applied and tested on an aquaglyceroporin channel through a series of simulations totaling 5 ns in duration. PMID- 12080110 TI - Water secretion associated with exocytosis in endocrine cells revealed by micro forcemetry and evanescent wave microscopy. AB - It has been a long belief that release of substances from the cell to the extracellular milieu by exocytosis is completed by diffusion of the substances from secretory vesicles through the fusion pore. Involvement of any mechanical force that may be superposed on the diffusion to enhance the releasing process has not been elucidated to date. We tackled this problem in cultured bovine chromaffin cells using direct and sensitive methods: the laser-trap forcemetry and the evanescent-wave fluorescence microscopy. With a laser beam, we trapped a micro bead in the vicinity of a cell (with 1 microm of separation) and observed movements of the bead optically. Electrical stimulation of the cell induced many of rapid and transient movements of the bead in a direction away from the cell surface. Upon the same stimulation, secretory vesicles stained with a fluorescent probe, acridine orange, and excited under the evanescent field illumination, showed a flash-like response: a transient increase in fluorescence intensity associated with a diffuse cloud of brightness, followed by a complete disappearance. These mechanical and fluorescence transients indicate a directional flow of substances. Blockers of the Cl(-) channel suppressed the rates of both responses in a characteristic way but not exocytotic fusion itself. Immunocytochemical studies revealed the presence of Cl(-) and K(+) channels on the vesicle membranes. These results suggest that the externalization of hormones or transmitters upon exocytosis of vesicles is augmented by secretion of water from the vesicle membrane through the widened fusion pore, possibly modulating the rate and reach of the hormone or transmitter release and facilitating transport of the signal molecules in intercellular spaces. PMID- 12080111 TI - Molecular mechanism of spontaneous pigment activation in retinal cones. AB - Spontaneous current and voltage fluctuations (dark noise) in the photoreceptor cells of the retina limit the ability of the visual system to detect dim light. We recorded the dark current noise of individual salamander L cones. Previous work showed that the dark noise in these cells arises from thermal activation of the visual pigment. From the temperature dependence of the rate of occurrence of elementary noise events, we found an Arrhenius activation energy E(a) of 25 +/- 7 kcal/mol (mean +/- SD). This E(a) is similar to that reported for the thermal isomerization of 11-cis retinal in solution, suggesting that the cone pigment noise results from isomerization of the retinal chromophore. E(a) for the cone noise is similar to that previously reported for the "photon-like" noise of rods, but the preexponential factor is five orders of magnitude higher. To test the hypothesis that thermal isomerization can only occur in molecules whose Schiff base linkage is unprotonated, we changed the pH of the solution bathing the cone outer segment. This had little effect on the rate of occurrence of elementary noise events. The rate was also unchanged when the cone was exposed to Ringer solution made up from heavy water, whose solvent isotope effect should reduce the probability, that the Schiff base nitrogen is naked. PMID- 12080113 TI - Early fluorescence signals detect transitions at mammalian serotonin transporters. AB - The mammalian serotonin transporters rSERT or hSERT were expressed in oocytes and labeled with sulforhodamine-MTS. The endogenous Cys-109 residue contributes most of the signal, and the labeled transporter shows normal function. The SERT fluorescence decreases in the presence of 5-HT and also depends on the inorganic substrates of SERT. The fluorescence also increases with membrane depolarization. During voltage-jump experiments, fluorescence relaxations show little inactivation or history dependence. The fluorescence signal has a voltage dependence similar to that of the prepriming step of the previously described voltage-dependent transient current. However, the fluorescence relaxations are the fastest voltage-dependent events yet studied at SERT; their time constants of approximately 8-30 ms are severalfold faster than the prepriming or inactivation phases of the transient currents. These fluorescence signals are interpreted within the framework of the gate-lumen-gate model. The signals may monitor initial events at the outer gate. PMID- 12080112 TI - Selectin-like kinetics and biomechanics promote rapid platelet adhesion in flow: the GPIb(alpha)-vWF tether bond. AB - The ability of platelets to tether to and translocate on injured vascular endothelium relies on the interaction between the platelet glycoprotein receptor Ib alpha (GPIb(alpha)) and the A1 domain of von Willebrand factor (vWF-A1). To date, limited information exists on the kinetics that govern platelet interactions with vWF in hemodynamic flow. We now report that the GPIb(alpha)-vWF A1 tether bond displays similar kinetic attributes as the selectins including: 1) the requirement for a critical level of hydrodynamic flow to initiate adhesion, 2) short-lived tethering events at sites of vascular injury in vivo, and 3) a fast intrinsic dissociation rate constant, k(0)(off) (3.45 +/- 0.37 s(-1)). Values for k(off), as determined by pause time analysis of transient capture/release events, were also found to vary exponentially (4.2 +/- 0.8 s(-1) to 7.3 +/- 0.4 s(-1)) as a function of the force applied to the bond (from 36 to 217 pN). The biological importance of rapid bond dissociation in platelet adhesion is demonstrated by kinetic characterization of the A1 domain mutation, I546V that is associated with type 2B von Willebrand disease (vWD), a bleeding disorder that is due to the spontaneous binding of plasma vWF to circulating platelets. This mutation resulted in a loss of the shear threshold phenomenon, a approximately sixfold reduction in k(off), but no significant alteration in the ability of the tether bond to resist shear-induced forces. Thus, flow dependent adhesion and rapid and force-dependent kinetic properties are the predominant features of the GPIb(alpha)-vWF-A1 tether bond that in part may explain the preferential binding of platelets to vWF at sites of vascular injury, the lack of spontaneous platelet aggregation in circulating blood, and a mechanism to limit thrombus formation. PMID- 12080114 TI - Modifications of alamethicin ion channels by substitution of Glu-7 for Gln-7. AB - To evaluate the role of charged residues facing a pore lumen in stability of channel structure and ion permeation, we studied electrical properties of ion channels formed by synthesized native alamethicins (Rf50 (alm-Q7Q18) and Rf30 (alm-Q7E18)) and their analogs with Glu-7 (alm-E7Q18 and alm-E7E18). The single channel currents were measured over a pH range of 3.5 to 8.7 using planar bilayers of diphytanoyl PC. The peptides all showed multi-level current fluctuations in this pH range. At pH 3.5 the channels formed by the four peptides were similar to each other irrespective of the side chain differences at positions 7 and 18. The ionization of Glu-7 (E7) and Glu-18 (E18) above neutral pH reduced the relative probabilities of low-conductance states (levels 1 and 2) and increased those of high-conductance states (levels 4-6). The channel conductance of the peptides with E7 and/or E18, which was distinct from that of alm-Q7Q18, showed a marked pH-dependence, especially for low-conductance states. The ionization of E7 further reduced the stability of channel structure, altered the current-voltage curve from a superlinear relation to a sublinear one, and enhanced cation selectivity. These results indicate that ionized E7 strongly influences the channel structure and the ion permeation, in contrast to ionized E18. PMID- 12080115 TI - Cloning and expression of the human T-type channel Ca(v)3.3: insights into prepulse facilitation. AB - The full-length human Ca(v)3.3 (alpha(1I)) T-type channel was cloned, and found to be longer than previously reported. Comparison of the cDNA sequence to the human genomic sequence indicates the presence of an additional 4-kb exon that adds 214 amino acids to the carboxyl terminus and encodes the 3' untranslated region. The electrophysiological properties of the full-length channel were studied after transient transfection into 293 human embryonic kidney cells using 5 mM Ca(2+) as charge carrier. From a holding potential of -100 mV, step depolarizations elicited inward currents with an apparent threshold of -70 mV, a peak of -30 mV, and reversed at +40 mV. The kinetics of channel activation, inactivation, deactivation, and recovery from inactivation were very similar to those reported previously for rat Ca(v)3.3. Similar voltage-dependent gating and kinetics were found for truncated versions of human Ca(v)3.3, which lack either 118 or 288 of the 490 amino acids that compose the carboxyl terminus. A major difference between these constructs was that the full-length isoform generated twofold more current. These results suggest that sequences in the distal portion of Ca(v)3.3 play a role in channel expression. Studies on the voltage-dependence of activation revealed that a fraction of channels did not gate as low voltage activated channels, requiring stronger depolarizations to open. A strong depolarizing prepulse (+100 mV, 200 ms) increased the fraction of channels that gated at low voltages. In contrast, human Ca(v)3.3 isoforms with shorter carboxyl termini were less affected by a prepulse. Therefore, Ca(v)3.3 is similar to high voltage-activated Ca(2+) channels in that depolarizing prepulses can regulate their activity, and their carboxy termini play a role in modulating channel activity. PMID- 12080116 TI - Thermodynamically irreversible gating of ryanodine receptors in situ revealed by stereotyped duration of release in Ca(2+) sparks. AB - For a single or a group of Markov channels gating reversibly, distributions of open and closed times should be the sum of positively weighted decaying exponentials. Violation of this microscopic reversibility has been demonstrated previously on a number of occasions at the single channel level, and has been attributed to possible channel coupling to external sources of free energy. Here we show that distribution of durations of Ca(2+) release underlying Ca(2+) sparks in intact cardiac myocytes exhibits a prominent mode at approximately 8 ms. Analysis of the cycle time for repetitive sparks at hyperactive sites revealed no intervals briefer than approximately 35 ms and a mode at approximately 90 ms. These results indicate that, regardless of whether Ca(2+) sparks are single channel or multi-channel in origin, they are generated by thermodynamically irreversible stochastic processes. In contrast, data from planar lipid bilayer experiments were consistent with reversible gating of RyR under asymmetric cis (4 microM) and trans Ca(2+) (10 mM), suggesting that the irreversibility for Ca(2+) spark genesis may reside at a supramolecular level. Modeling suggests that Ca(2+) induced Ca(2+) release among adjacent RyRs may couple the external energy derived from Ca(2+) gradients across the SR to RyR gating in situ, and drive the irreversible generation of Ca(2+) sparks. PMID- 12080118 TI - Modeling diverse range of potassium channels with Brownian dynamics. AB - Using the experimentally determined KcsA structure as a template, we propose a plausible explanation for the diversity of potassium channels seen in nature. A simplified model of KcsA is constructed from its atomic resolution structure by smoothing out the protein-water boundary and representing the atoms forming the channel protein as a homogeneous, low dielectric medium. The properties of the simplified and atomic-detail models, deduced from electrostatic calculations and Brownian dynamics simulations, are shown to be qualitatively similar. We then study how the current flowing across the simplified model channel changes as the shape of the intrapore region is modified. This is achieved by increasing the radius of the intracellular pore systematically from 1.5 to 5 A while leaving the dimensions of the selectivity filter and inner chamber unaltered. The strengths of the dipoles located near the entrances of the channel, the carbonyl groups lining the selectivity filter, and the helix macrodipoles are kept constant. The channel conductance increases steadily as the radius of the intracellular pore is increased. The rate-limiting step for both the outward and inward current is the time it takes for an ion to cross the residual energy barrier located in the intrapore region. The current-voltage relationship obtained with symmetrical solutions is linear when the applied potential is less than approximately 100 mV but deviates slightly from Ohm's law at higher applied potentials. The nonlinearity in the current-voltage curve becomes less pronounced as the radius of the intracellular pore is increased. When the strengths of the dipoles near the intracellular entrance are reduced, the channel shows a pronounced inward rectification. Finally, the conductance exhibits the saturation property observed experimentally. We discuss the implications of these findings on the transport of ions across the potassium channels and membrane channels in general. PMID- 12080117 TI - NMR structures of the second transmembrane domain of the human glycine receptor alpha(1) subunit: model of pore architecture and channel gating. AB - Glycine receptors (GlyR) are the primary inhibitory receptors in the spinal cord and belong to a superfamily of ligand-gated ion channels (LGICs) that are extremely sensitive to low-affinity neurological agents such as general anesthetics and alcohols. The high-resolution pore architecture and the gating mechanism of this superfamily, however, remain unclear. The pore-lining second transmembrane (TM2) segments of the GlyR alpha(1) subunit are unique in that they form functional homopentameric channels with conductance characteristics nearly identical to those of an authentic receptor (Opella, S. J., J. Gesell, A. R. Valente, F. M. Marassi, M. Oblatt-Montal, W. Sun, A. F. Montiel, and M. Montal. 1997. Chemtracts Biochem. Mol. Biol. 10:153-174). Using NMR and circular dichroism (CD), we determined the high-resolution structures of the TM2 segment of human alpha(1) GlyR and an anesthetic-insensitive mutant (S267Y) in dodecyl phosphocholine (DPC) and sodium dodecyl sulfate (SDS) micelles. The NMR structures showed right-handed alpha-helices without kinks. A well-defined hydrophilic path, composed of side chains of G2', T6', T10', Q14', and S18', runs along the helical surfaces at an angle approximately 10-20 degrees relative to the long axis of the helices. The side-chain arrangement of the NMR-derived structures and the energy minimization of a homopentameric TM2 channel in a fully hydrated DMPC membrane using large-scale computation suggest a model of pore architecture in which simultaneous tilting movements of entire TM2 helices by a mere 10 degrees may be sufficient to account for the channel gating. The model also suggests that additional residues accessible from within the pore include L3', T7', T13', and G17'. A similar pore architecture and gating mechanism may apply to other channels in the same superfamily, including GABA(A), nACh, and 5 HT(3) receptors. PMID- 12080119 TI - A large-conductance anion channel of the Golgi complex. AB - An acidic lumenal pH is vital for the proper posttranslational modifications and sorting of proteins and lipids from the Golgi complex. We characterized ion channels present in Golgi fractions that have been cleared of transiting proteins. A large conductance anion channel was observed in approximately 30% of successful channel incorporations into the planar lipid bilayer. The channel, GOLAC-2, has six levels (one closed and five open). The open states are each approximately 20% increments of the maximal, 325 pS conductance. The channel was approximately 6 times more selective for Cl(-) over K(+). Binomial analysis of percent occupancy for each conducting level supports the hypothesis of five independent conducting pathways. The conducting levels can coordinately gate because full openings and closings were often observed. Addition of 3 to 5 mM reduced glutathione to the cis chamber caused dose-dependent increases in single channel conductance, indicating that the channel may be regulated by the oxidation-reduction state of the cell. We propose that GOLAC-2 is a co-channel complex consisting of five identical pores that have a coordinated gating mechanism. GOALC-2 may function as a source of counter anions for the H(+)-ATPase and may be involved in regulating charge balance and membrane potential of the Golgi complex. PMID- 12080120 TI - Purification of the small mechanosensitive channel of Escherichia coli (MscS): the subunit structure, conduction, and gating characteristics in liposomes. AB - The small mechanosensitive channel, MscS, is a part of the turgor-driven solute efflux system that protects bacteria from lysis in the event of osmotic downshift. It has been identified in Escherichia coli as a product of the orphan yggB gene, now called mscS (Levina et al., 1999, EMBO J. 18:1730). Here I show that that the isolated 31-kDa MscS protein is sufficient to form a functional mechanosensitive channel gated directly by tension in the lipid bilayer. MscS 6His complexes purified in the presence of octylglucoside and lipids migrate in a high-resolution gel-filtration column as particles of approximately 200 kDa. Consistent with that, the protein cross-linking patterns predict a hexamer. The channel reconstituted in soybean asolectin liposomes was activated by pressures of 20-60 mm Hg and displayed the same asymmetric I-V curve and slight anionic preference as in situ. At the same time, the single-channel conductance is proportional to the buffer conductivity in a wide range of salt concentrations. The rate of channel activation in response to increasing pressure gradient across the patch was slower than the rate of closure in response to decreasing steps of pressure gradient. Therefore, the open probability curves were recorded with descending series of pressures. Determination of the curvature of patches by video imaging permitted measurements of the channel activity as a function of membrane tension (gamma). Po(gamma) curves had the midpoint at 5.5 +/- 0.1 dyne/cm and gave estimates for the energy of opening DeltaG = 11.4 +/- 0.5 kT, and the transition-related area change DeltaA = 8.4 +/- 0.4 nm(2) when fitted with a two-state Boltzmann model. The correspondence between channel properties in the native and reconstituted systems is discussed. PMID- 12080121 TI - Adhesion of nanoparticles to vesicles: a Brownian dynamics simulation. AB - We studied the interaction of bilayer vesicles and adhesive nanoparticles using a Brownian dynamics simulation. The nanoparticles are simple models of proteins or colloids. The adhering nanoparticle induces the morphological change of the vesicle: budding, formation of two vesicles in which only outer monolayers are connected, and fission. We also show that the nanoparticle promotes the fusion process: fusion-pore opening from a stalk intermediate, a neck-like structure that only connects outer monolayers of two vesicles. The nanoparticle bends the stalk, and induces the pore opening. PMID- 12080122 TI - Site-directed mutagenesis of tyrosine 118 within the central constriction site of the LamB (maltoporin) channel of Escherichia coli. II. Effect on maltose and maltooligosaccharide binding kinetics. AB - The 3-D structure of the maltooligosaccharide-specific LamB channel of Escherichia coli (also called maltoporin) is known from x-ray crystallography. The central constriction of the channel formed by the external loop 3 is controlled by tyrosine 118. Y118 was replaced by site-directed mutagenesis by 10 other amino acids (alanine (A), isoleucine (I), asparagine (N), serine (S), cysteine (C), aspartic acid (D), arginine (R), histidine (H), phenylalanine (F), and tryptophan (W)) including neutral ones, negatively and positively charged amino acids to study the effect of their size, their hydrophobicity index, and their charge on maltose and maltooligosaccharide binding to LamB. The mutants were reconstituted into lipid bilayer membranes and the stability constants for binding of maltose, maltotriose, maltopentaose, and maltoheptaose to the channel were measured using titration experiments. The mutation of Y118 to any other non aromatic amino acid led to a substantial decrease of the stability constant of binding by factors between about two and six. The highest effect was observed for the mutant Y118A. Replacement of Y118 by the two other aromatic amino acids, phenylalanine (F) and tryptophan (W), resulted in a substantial increase of the stability constant maximally by a factor of almost 400 for the Y118W mutant. The carbohydrate-induced block of the channel function was used for the study of current noise through the different mutant LamB channels. The analysis of the power density spectra allowed the evaluation of the on- and off-rate constants (k(1) and k(-1)) of sugar binding. The results suggest that both rate constants were affected by the mutations. For most mutants, with the exception of Y118F and Y118W, k(1) decreased and k(-1) increased, whereas the opposite was found for the aromatic amino acid mutants. The results suggest that tyrosine 118 has a crucial effect on carbohydrate transport through LamB. PMID- 12080123 TI - Biophysical characterization of triacyl monosaccharide lipid a partial structures in relation to bioactivity. AB - Synthetic triacyl glucosamine monosaccharide lipid A part structures corresponding to the non-reducing moiety of enterobacterial lipid A with an acyloxyacyl chain linked to position 3 of the glucosamine and an unbranched chain linked to position 2 (group 1) and vice versa (group 2) were analyzed biophysically: Fourier-transform infrared spectroscopy was performed to characterize the gel-to-liquid crystalline phase transition, the phosphate band contour, and the orientation of the glucosamine with respect to the membrane surface. Small-angle x-ray diffraction was applied for the elucidation of the supramolecular aggregate structure and, with that, of the molecular shape. With fluorescence resonance energy transfer the lipopolysaccharide-binding protein (LBP)-mediated intercalation of the lipid A partial structures into phospholipid liposomes was monitored. The physical data clearly exhibit a classification of the synthetic compounds into two groups: group 1 compounds have sharp phase transitions, indicating dense acyl chain packing and an inclination of the glucosamine backbone with respect to the membrane surface of 30 degrees with the phosphate buried in the membrane. Group 2 compounds have a very broad phase transition, indicating poorly packed acyl chains, and an inclination of -30 degrees with the phosphate group sticking outward. For the first group unilamellar phases are observed superimposed by a non-lamellar structure, and for the second one only multilamellar aggregate structures. The cytokine-inducing capacity in human mononuclear cells is relatively high for the first group and low or absent for the second group. Based on these data a model of the intra and intermolecular conformations is proposed which also extends the concept of "endotoxic conformation." PMID- 12080124 TI - Modulation of concentration fluctuations in phase-separated lipid membranes by polypeptide insertion. AB - The lateral membrane organization and phase behavior of the binary lipid mixture DMPC (1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine) - DSPC (1,2-distearoyl-sn glycero-3-phosphatidylcholine) without and with incorporated gramicidin D (GD) as a model biomembrane polypeptide was studied by small-angle neutron scattering, Fourier-transform infrared spectroscopy, and by two-photon excitation fluorescence microscopy on giant unilamellar vesicles. The small-angle neutron scattering method allows the detection of concentration fluctuations in the range from 1 to 200 nm. Fluorescence microscopy was used for direct visualization of the lateral lipid organization and domain shapes on a micrometer length scale including information of the lipid phase state. In the fluid-gel coexistence region of the pure binary lipid system, large-scale concentration fluctuations appear. Infrared spectral parameters were used to determine the peptide conformation adopted in the different lipid phases. The data show that the structure of the temperature-dependent lipid phases is significantly altered by the insertion of 2 to 5 mol% GD. At temperatures corresponding to the gel-fluid phase coexistence region the concentration fluctuations drastically decrease, and we observe domains in the giant unilamellar vesicles, which mainly disappear by the incorporation of 2 to 5 mol% GD. Further, the lipid matrix has the ability to modulate the conformation of the inserted polypeptide. The balance between double helical and helical dimer structures of GD depends on the phospholipid chain length and phase state. A large hydrophobic mismatch, such as in gel phase one component DSPC bilayers, leads to an increase in population of double-helical structures. Using an effective molecular sorting mechanism, a large hydrophobic mismatch can be avoided in the DMPC-DSPC lipid mixture, which leads to significant changes in the heterogeneous lipid structure and in polypeptide conformation. PMID- 12080125 TI - Organization of model helical peptides in lipid bilayers: insight into the behavior of single-span protein transmembrane domains. AB - Selectively deuterated transmembrane peptides comprising alternating leucine alanine subunits were examined in fluid bilayer membranes by solid-state nuclear magnetic resonance (NMR) spectroscopy in an effort to gain insight into the behavior of membrane proteins. Two groups of peptides were studied: 21-mers having a 17-amino-acid hydrophobic domain calculated to be close in length to the hydrophobic thickness of 1-palmitoyl-2-oleoyl phosphatidylcholine and 26-mers having a 22-amino-acid hydrophobic domain calculated to exceed the membrane hydrophobic thickness. (2)H NMR spectral features similar to ones observed for transmembrane peptides from single-span receptors of higher animal cells were identified which apparently correspond to effectively monomeric peptide. Spectral observations suggested significant distortion of the transmembrane alpha-helix, and/or potential for restriction of rotation about the tilted helix long axis for even simple peptides. Quadrupole splittings arising from the 26-mer were consistent with greater peptide "tilt" than were those of the analogous 21-mer. Quadrupole splittings associated with monomeric peptide were relatively insensitive to concentration and temperature over the range studied, indicating stable average conformations, and a well-ordered rotation axis. At high peptide concentration (6 mol% relative to phospholipid) it appeared that the peptide predicted to be longer than the membrane thickness had a particular tendency toward reversible peptide-peptide interactions occurring on a timescale comparable with or faster than approximately 10(-5) s. This interaction may be direct or lipid-mediated and was manifest as line broadening. Peptide rotational diffusion rates within the membrane, calculated from quadrupolar relaxation times, T(2e), were consistent with such interactions. In the case of the peptide predicted to be equal to the membrane thickness, at low peptide concentration spectral lineshape indicated the additional presence of a population of peptide having rotational motion that was restricted on a timescale of 10(-5) s. PMID- 12080127 TI - Rotation of periphery methylpyridine of meso-tetrakis(n-N methylpyridiniumyl)porphyrin (n = 2, 3, 4) and its selective binding to native and synthetic DNAs. AB - By utilizing circular and linear dichroism, the binding mode of meso-tetrakis(n-N methylpyridiniumyl)porphyrin (n = 2, 3, 4) to various DNAs was studied in this work. 2-N-(methylpyridiniumyl)porphyrin(o-TMPyP), in which rotation of the periphery pyridinium ring is prevented, exhibits similar spectral properties when bound to DNA, poly[d(G-C)(2)] and poly[d(A-T)(2)], suggesting a similar binding mode. Close analysis of the spectral properties led us to conclude that o-TMPyP sits in the major groove. However, both 3-N- and 4-N (methylpyridiniumyl)porphyrin (m- and p-TMPyP), of which the periphery pyridinium ring is free to rotate, intercalate between the basepairs of DNA and poly[d(G C)(2)]. In the presence of poly[d(A-T)(2)], m-TMPyP exhibits a typical bisignate excitonic CD spectrum in the Soret band, while p-TMPyP shows two positive CD bands. The excitonic CD spectrum of the m-TMPyP-poly[d(A-T)(2)] complex and the positive CD band of the o-TMPyP-poly[d(A-T)(2)] complex were not affected by the presence of the minor groove binding drug, 4',6-diamidino-2-phenylindole (DAPI), indicating that this porphyrin is bound in the major groove. In contrast, two positive CD bands of the p-TMPyP-poly[d(A-T)(2)] complex altered in the presence of DAPI. From the changes in CD spectrum and other spectral properties, a few possible binding modes for p-TMPyP to poly[d(A-T)(2)] are suggested. PMID- 12080126 TI - Kinetic effects of myosin regulatory light chain phosphorylation on skeletal muscle contraction. AB - Kinetic analysis of contracting fast and slow rabbit muscle fibers in the presence of the tension inhibitor 2,3-butanedione monoxime suggests that regulatory light chain (RLC) phosphorylation up-regulates the flux of weakly attached cross-bridges entering the contractile cycle by increasing the actin catalyzed release of phosphate from myosin. This step appears to be separate from earlier Ca(2+) regulated steps. Small step-stretches of single skinned fibers were used to study the effect of phosphorylation on fiber mechanics. Subdivision of the resultant tension transients into the Huxley-Simmons phases 1, 2(fast), 2(slow), 3, and 4 reveals that phosphorylation reduces the normalized amplitude of the delayed rise in tension (stretch activation response) by decreasing the amplitudes of phase 3 and, to a lesser extent, phase 2(slow). In slow fibers, the RLC P1 isoform phosphorylates at least 4-fold faster than the P2 isoform, complicating the role of RLC phosphorylation in heart and slow muscle. We discuss the functional relevance of the regulation of stretch activation by RLC phosphorylation for cardiac and other oscillating muscles and speculate how the interaction of the two heads of myosin could account for the inverse effect of Ca(2+) levels on isometric tension and rate of force redevelopment (k(TR)). PMID- 12080129 TI - Spectroscopy of single phycoerythrocyanin monomers: dark state identification and observation of energy transfer heterogeneities. AB - Phycoerythrocyanin (PEC) is part of the light harvesting system of cyanobacteria. The PEC monomer contains one phycoviolobilin chromophore, which transfers excitation energy onto two phycocyanobilin chromophores. Many spectroscopical methods have been used in the past to study the bulk properties of PEC. These methods average over many molecules. Therefore, differences in the behavior of individual molecules remain hidden. The energy transfer within photosynthetic complexes is however sensitive to changes in the spectroscopic properties of the participating subunits. Knowledge about heterogeneities is therefore important for the description of the energy transfer in photosynthetic systems. Here, the recording of the fluorescence emission of single PEC molecules is used as a tool to obtain such information. Spectrally resolved detection as well as double resonance excitation of single PEC molecules is used to investigate their bleaching behavior. The trans isomer of the phycoviolobilin chromophore is identified as a short-lived dark state of monomeric PEC. Polarization sensitive single molecule detection is used for the direct observation of the energy transfer in individual PEC molecules. The experiments reveal that more than one half of the PEC molecules exhibit an energy transfer behavior significantly different from the bulk. These heterogeneities persist on a time scale of several seconds. Model calculations lead to the conclusion that they are caused by minor shifts in the spectra of the chromophores. PMID- 12080128 TI - Accurate representation of B-DNA double helical structure with implicit solvent and counterions. AB - High-resolution nuclear magnetic resonance (NMR) and crystallographic data have been taken to refine the force field used in the torsion angle space nucleic acids molecular mechanics program DUPLEX. The population balance deduced from NMR studies of two carcinogen-modified DNA conformers in equilibrium was used to fine tune a sigmoidal, distance-dependent dielectric function so that reasonable relative energies could be obtained. In addition, the base-pair and backbone geometry from high-resolution crystal structures of the Dickerson-Drew dodecamer was used to re-evaluate the deoxyribose pseudorotation profile and the Lennard Jones nonbonded energy terms. With a modified dielectric function that assumes a very steep distance-dependent form, a deoxyribose pseudorotation profile with reduced energy barriers between C2'- and C3'-endo minima, and a shift of the Lennard-Jones potential energy minimum to a distance approximately 0.4 A greater than the sum of the van der Waals' radii, the sequence-dependent conformational features of the Dickerson-Drew dodecamer in both the solid state and the aqueous liquid crystalline phase are well reproduced. The robust performance of the revised force field, in conjunction with its efficiency through implicit treatment of solvent and counterions, provides a valuable tool for elucidating conformations and structure-function relationships of DNA, including those of molecules modified by carcinogens and other ligands. PMID- 12080130 TI - Proton transfer dynamics on the surface of the late M state of bacteriorhodopsin. AB - The cytoplasmic surface of the BR (initial) state of bacteriorhodopsin is characterized by a cluster of three carboxylates that function as a proton collecting antenna. Systematic replacement of most of the surface carboxylates indicated that the cluster is made of D104, E161, and E234 (Checover, S., Y. Marantz, E. Nachliel, M. Gutman, M. Pfeiffer, J. Tittor, D. Oesterhelt, and N. Dencher. 2001. Biochemistry. 40:4281-4292), yet the BR state is a resting configuration; thus, its proton-collecting antenna can only indicate the presence of its role in the photo-intermediates where the protein is re-protonated by protons coming from the cytoplasmic matrix. In the present study we used the D96N and the triple (D96G/F171C/F219L) mutant for monitoring the proton-collecting properties of the protein in its late M state. The protein was maintained in a steady M state by continuous illumination and subjected to reversible pulse protonation caused by repeated excitation of pyranine present in the reaction mixture. The re-protonation dynamics of the pyranine anion was subjected to kinetic analysis, and the rate constants of the reaction of free protons with the surface groups and the proton exchange reactions between them were calculated. The reconstruction of the experimental signal indicated that the late M state of bacteriorhodopsin exhibits an efficient mechanism of proton delivery to the unoccupied-most basic-residue on its cytoplasmic surface (D38), which exceeds that of the BR configuration of the protein. The kinetic analysis was carried out in conjunction with the published structure of the M state (Sass, H., G. Buldt, R. Gessenich, D. Hehn, D. Neff, R. Schlesinger, J. Berendzen, and P. Ormos. 2000. Nature. 406:649-653), the model that resolves most of the cytoplasmic surface. The combination of the kinetic analysis and the structural information led to identification of two proton-conducting tracks on the protein's surface that are funneling protons to D38. One track is made of the carboxylate moieties of residues D36 and E237, while the other is made of D102 and E232. In the late M state the carboxylates of both tracks are closer to D38 than in the BR (initial) state, accounting for a more efficient proton equilibration between the bulk and the protein's proton entrance channel. The triple mutant resembles in the kinetic properties of its proton conducting surface more the BR-M state than the initial state confirming structural similarities with the BR-M state and differences to the BR initial state. PMID- 12080131 TI - Tyr-199 and charged residues of pharaonis Phoborhodopsin are important for the interaction with its transducer. AB - pharaonis Phoborhodopsin (ppR; also pharaonis sensory rhodopsin II, psRII) is a retinal protein in Natronobacterium pharaonis and is a receptor of negative phototaxis. It forms a complex with its transducer, pHtrII, in membranes and transmits light signals by protein-protein interaction. Tyr-199 is conserved completely in phoborhodopsins among a variety of archaea, but it is replaced by Val (for bacteriorhodopsin) and Phe (for sensory rhodopsin I). Previously, we (Sudo, Y., M. Iwamoto, K. Shimono, and N. Kamo, submitted for publication) showed that analysis of flash-photolysis data of a complex between D75N and the truncated pHtrII (t-Htr) give a good estimate of the dissociation constant K(D) in the dark. To investigate the importance of Tyr-199, K(D) of double mutants of D75N/Y199F or D75N/Y199V with t-Htr was estimated by flash-photolysis and was approximately 10-fold larger than that of D75N, showing the significant contribution of Tyr-199 to binding. The K(D) of the D75N/t-Htr complex increased with decreasing pH, and the data fitted well with the Henderson-Hasselbach equation with a single pK(a) of 3.86 +/- 0.02. This suggests that certain deprotonated carboxyls at the surface of the transducer (possibly Asp-102, Asp 104, and Asp-106) are needed for the binding. PMID- 12080132 TI - Light harvesting in photosystem I: modeling based on the 2.5-A structure of photosystem I from Synechococcus elongatus. AB - The structure of photosystem I from the thermophilic cyanobacterium Synechococcus elongatus has been recently resolved by x-ray crystallography to 2.5-A resolution. Besides the reaction center, photosystem I consists also of a core antenna containing 90 chlorophyll and 22 carotenoid molecules. It is their function to harvest solar energy and to transfer this energy to the reaction center (RC) where the excitation energy is converted into a charge separated state. Methods of steady-state optical spectroscopy such as absorption, linear, and circular dichroism have been applied to obtain information on the spectral properties of the complex, whereas transient absorption and fluorescence studies reported in the literature provide information on the dynamics of the excitation energy transfer. On the basis of the structure, the spectral properties and the energy transfer kinetics are simultaneously modeled by application of excitonic coupling theory to reveal relationships between structure and function. A spectral assignment of the 96 chlorophylls is suggested that allows us to reproduce both optical spectra and transfer and emission spectra and lifetimes of the photosystem I complex from S. elongatus. The model calculation allowed to study the influence of the following parameters on the excited state dynamics: the orientation factor, the heterogeneous site energies, the modifications arising from excitonic coupling (redistribution of oscillator strength, energetic splitting, reorientation of transition dipoles), and presence or absence of the linker cluster chlorophylls between antenna and reaction center. For the Forster radius and the intrinsic primary charge separation rate, the following values have been obtained: R(0) = 7.8 nm and k(CS) = 0.9 ps(-1). Variations of these parameters indicate that the excited state dynamics is neither pure trap limited, nor pure transfer (to-the-trap) limited but seems to be rather balanced. PMID- 12080133 TI - The effect of core destabilization on the mechanical resistance of I27. AB - It is still unclear whether mechanical unfolding probes the same pathways as chemical denaturation. To address this point, we have constructed a concatamer of five mutant I27 domains (denoted (I27)(5)*) and used it for mechanical unfolding studies. This protein consists of four copies of the mutant C47S, C63S I27 and a single copy of C63S I27. These mutations severely destabilize I27 (DeltaDeltaG(UN) = 8.7 and 17.9 kJ mol(-1) for C63S I27 and C47S, C63S I27, respectively). Both mutations maintain the hydrogen bond network between the A' and G strands postulated to be the major region of mechanical resistance for I27. Measuring the speed dependence of the force required to unfold (I27)(5)* in triplicate using the atomic force microscope allowed a reliable assessment of the intrinsic unfolding rate constant of the protein to be obtained (2.0 x 10(-3) s( 1)). The rate constant of unfolding measured by chemical denaturation is over fivefold faster (1.1 x 10(-2) s(-1)), suggesting that these techniques probe different unfolding pathways. Also, by comparing the parameters obtained from the mechanical unfolding of a wild-type I27 concatamer with that of (I27)(5)*, we show that although the observed forces are considerably lower, core destabilization has little effect on determining the mechanical sensitivity of this domain. PMID- 12080134 TI - What causes hyperfluorescence: folding intermediates or conformationally flexible native states? AB - Hyperfluorescent intensity maxima during protein unfolding titrations are often taken as a sign for a thermodynamic folding intermediate. Here we explore another possibility: that hyperfluorescence could be the signature of a "pretransition" conformationally loosened native state. To model such native states, we study mutants of a fluorescent ubiquitin variant, placing cavities at various distances from the tryptophan fluorophore. We examine the correlation between protein flexibility and enhanced fluorescence intensity by using circular dichroism, fluorescence intensity unfolding titrations, fluorescence anisotropy measurements, and molecular dynamics. Based on experiment and simulation, we propose a simple model for hyperfluorescence in terms of static and dynamic conformational properties of the native state during unfolding. Apomyoglobin denaturant unfolding and phosphoglycerate kinase cold denaturation are discussed as examples. Our results do not preclude the existence of thermodynamic intermediates but do raise caution that by itself, hyperfluorescence during unfolding titrations is not conclusive proof of thermodynamic folding intermediates. PMID- 12080135 TI - Structure of the 1-36 N-terminal fragment of human phospholamban phosphorylated at Ser-16 and Thr-17. AB - The structure of a 36-amino-acid-long N-terminal fragment of human phospholamban phosphorylated at Ser-16 and Thr-17 and Cys-36-->Ser mutated was determined from nuclear magnetic resonance data in aqueous solution containing 30% trifluoroethanol. The peptide assumes a conformation characterized by two alpha helices connected by an irregular strand, which comprises the amino acids from Arg-13 to Pro-21. The proline is in a trans conformation. The two phosphate groups on Ser-16 and Thr-17 are shown to interact preferably with the side chains of Arg-14 and Arg-13, respectively. The helix comprising amino acids 22 to 35 is well determined (the rmsd for the backbone atoms, calculated for a family of 24 nuclear magnetic resonance structures is 0.69 +/- 0.28 A). The structures of phosphorylated and unphosphorylated phospholamban are compared, and the effect of the two phosphate groups on the relative spatial position of the two helices is examined. The packing parameters Omega (interhelical angle) and d (minimal interhelical distance) are calculated: in the case of the phosphorylated phospholamban, Omega = 100 +/- 35 degrees and d = 7.9 +/- 4.6 A, whereas for the unphosphorylated peptide the values are Omega = 80 +/- 20 degrees and d = 7.0 +/- 4.0 A. We conclude that 1) the phosphorylation does not affect the structure of the C terminus between residues 21 and 36 and 2) the phosphorylated phospholamban has more loose helical packing than the nonphosphorylated. PMID- 12080136 TI - An automated two-dimensional optical force clamp for single molecule studies. AB - We constructed a next-generation optical trapping instrument to study the motility of single motor proteins, such as kinesin moving along a microtubule. The instrument can be operated as a two-dimensional force clamp, applying loads of fixed magnitude and direction to motor-coated microscopic beads moving in vitro. Flexibility and automation in experimental design are achieved by computer control of both the trap position, via acousto-optic deflectors, and the sample position, using a three-dimensional piezo stage. Each measurement is preceded by an initialization sequence, which includes adjustment of bead height relative to the coverslip using a variant of optical force microscopy (to +/-4 nm), a two dimensional raster scan to calibrate position detector response, and adjustment of bead lateral position relative to the microtubule substrate (to +/-3 nm). During motor-driven movement, both the trap and stage are moved dynamically to apply constant force while keeping the trapped bead within the calibrated range of the detector. We present details of force clamp operation and preliminary data showing kinesin motor movement subject to diagonal and forward loads. PMID- 12080137 TI - Laser-scanning coherent anti-Stokes Raman scattering microscopy and applications to cell biology. AB - Laser-scanning coherent anti-Stokes Raman scattering (CARS) microscopy with fast data acquisition and high sensitivity has been developed for vibrational imaging of live cells. High three-dimensional (3D) resolution is achieved with two collinearly overlapped near infrared picosecond beams and a water objective with a high numerical aperture. Forward-detected CARS (F-CARS) and epi-detected CARS (E-CARS) images are recorded simultaneously. F-CARS is used for visualizing features comparable to or larger than the excitation wavelength, while E-CARS allows detection of smaller features with a high contrast. F-CARS and E-CARS images of live and unstained cells reveal details invisible in differential interference-contrast images. High-speed vibrational imaging of unstained cells undergoing mitosis and apoptosis has been carried out. For live NIH 3T3 cells in metaphase, 3D distribution of chromosomes is mapped at the frequency of the DNA backbone Raman band, while the vesicles surrounding the nucleus is imaged by E CARS at the frequency of the C-H stretching Raman band. Apoptosis in NIH 3T3 cells is monitored using the CARS signal from aliphatic C-H stretching vibration. PMID- 12080138 TI - Anisotropic diffusion in mitral cell dendrites revealed by fluorescence correlation spectroscopy. AB - Fluorescence correlation spectroscopy (FCS) can be used to measure kinetic properties of single molecules in drops of solution or in cells. Here we report on FCS measurements of tetramethylrhodamine (TMR)-dextran (10 kDa) in dendrites of cultured mitral cells of Xenopus laevis tadpoles. To interpret such measurements correctly, the plasma membrane as a boundary of diffusion has to be taken into account. We show that the fluorescence data recorded from dendrites are best described by a model of anisotropic diffusion. As compared to diffusion in water, diffusion of the 10-kDa TMR-dextran along the dendrite is slowed down by a factor 1.1-2.1, whereas diffusion in lateral direction is 10-100 times slower. The dense intradendritic network of microtubules oriented parallel to the dendrite is discussed as a possible basis for the observed anisotropy. In somata, diffusion was found to be isotropic in three dimensions and 1.2-2.6 times slower than in water. PMID- 12080139 TI - Dihedral angles of tripeptides in solution directly determined by polarized Raman and FTIR spectroscopy. AB - The amide I mode of the peptide linkage is highly delocalized in peptides and protein segments due to through-bond and through-space vibrationally coupling between adjacent peptide groups. J. Phys. Chem. B. 104:11316-11320) used coherent femtosecond infrared (IR) spectroscopy to determine the excitonic coupling energy and the orientational angle between the transition dipole moments of the interacting amide I modes of cationic tri-alanine in D(2)O. Recently, the same parameters were determined for all protonation states of tri-alanine by analyzing the amide I bands in the respective IR and isotropic Raman spectra (. J. Am. Chem. Soc. 119:1720-1726.). In both studies, the dihedral angles phi and psi were then obtained by utilizing the orientational dependence of the coupling energy obtained from ab initio calculations on tri-glycine in vacuo (. J. Raman Spectrosc. 29:81-86) to obtain an extended 3(1) helix-like structure for the tripeptide. In the present paper, a novel algorithm for the analysis of excitonic coupling between amide I modes is presented, which is based on the approach by Schweitzer-Stenner et al. but avoids the problematic use of results from ab initio calculations. Instead, the dihedral angles are directly determined from infrared and visible polarized Raman spectra. First, the interaction energy and the corresponding degree of wave-function mixing were obtained from the amide I profile in the isotropic Raman spectrum. Second, the depolarization ratios and the amide I profiles in the anisotropic Raman and IR-absorption spectra were used to determine the orientational angle between the peptide planes and the transition dipole moments, respectively. Finally, these two geometric parameters were utilized to determine the dihedral angles phi and psi between the interacting peptide groups. Stable extended conformations with dihedral angles in the beta-sheet region were obtained for all protonation states of tri-alanine, namely phi(+) = -126 degrees, psi(+) = 178 degrees; phi(+/-) = -110 degrees, psi(+/-) = 155 degrees; and phi(-) = -127 degrees, psi(-) = 165 degrees for the cationic, zwitterionic, and anionic state, respectively. These values reflect an extended beta-helix structure. Tri-glycine was found to be much more heterogeneous in that different extended conformers coexist in the cationic and zwitterionic state, which yield a noncoincidence between isotropic and anisotropic Raman scattering. Our study introduces vibrational spectroscopy as a suitable tool for the structure analysis of peptides in solution and tripeptides as suitable model systems for investigating the role of local interactions in determining the propensity of peptide segments for distinct secondary structure motifs. PMID- 12080140 TI - Analysis of coupled bimolecular reaction kinetics and diffusion by two-color fluorescence correlation spectroscopy: enhanced resolution of kinetics by resonance energy transfer. AB - In two-color fluorescence correlation spectroscopy (TCFCS), the fluorescence intensities of two fluorescently-labeled species are cross-correlated over time and can be used to identify static and dynamic interactions. Generally, fluorophore labels are chosen that do not undergo Forster resonance energy transfer (FRET). Here, a general TCFCS theory is presented that accounts for the possibility of FRET between reactants in the reversible bimolecular reaction, [reaction: see text] where k(f) and k(b) are forward and reverse rate constants, respectively (dissociation constant K(d) = k(b)/k(f)). Using this theory, we systematically investigated the influence on the correlation function of FRET, reaction rates, reactant concentrations, diffusion, and component visibility. For reactants of comparable size and an energy-transfer efficiency of approximately 90%, experimentally measurable cross-correlation functions should be sensitive to reaction kinetics for K(d) > 10(-8) M and k(f) >or= approximately 10(7) M(-1)s( 1). Measured auto-correlation functions corresponding to donor and acceptor labels are generally less sensitive to reaction kinetics, although for the acceptor, this sensitivity increases as the visibility of the donor increases relative to the acceptor. In the absence of FRET or a significant hydrodynamic difference between reactant species, there is little effect of reaction kinetics on the shape of auto- and cross-correlation functions. Our results suggest that a subset of biologically relevant association-dissociation kinetics can be measured by TCFCS and that FRET can be advantageous in enhancing these effects. PMID- 12080141 TI - Fluorescence light microscopy of pulmonary surfactant at the air-water interface of an air bubble of adjustable size. AB - The structural dynamics of pulmonary surfactant was studied by epifluorescence light microscopy at the air-water interface of a bubble as a model close to nature for an alveolus. Small unilamellar vesicles of dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylglycerol, a small amount of a fluorescent dipalmitoylphosphatidylcholine-analog, and surfactant-associated protein C were injected into the buffer solution. They aggregated to large clusters in the presence of Ca(2+) and adsorbed from these units to the interface. This gave rise to an interfacial film that eventually became fully condensed with dark, polygonal domains in a fluorescent matrix. When now the bubble size was increased or decreased, respectively, the film expanded or contracted. Upon expansion of the bubble, the dark areas became larger to the debit of the bright matrix and reversed upon contraction. We were able to observe single domains during the whole process. The film remained condensed, even when the interface was increased to twice its original size. From comparison with scanning force microscopy directly at the air-water interface, the fluorescent areas proved to be lipid bilayers associated with the (dark) monolayer. In the lung, such multilayer phase acts as a reservoir that guarantees a full molecular coverage of the alveolar interface during the breathing cycle and provides mechanical stability to the film. PMID- 12080142 TI - Thermodynamics of cationic lipid-DNA complex formation as studied by isothermal titration calorimetry. AB - The detailed analysis of the cationic lipid-DNA complex formation by means of isothermal titration calorimetry is presented. Most experiments were done using 1,2-dioleyl-sn-glycero-3-ethylphosphocholine (EDOPC), but basic titrations were also done using DOTAP, DOTAP:DOPC, and DOTAP:DOPE mixtures. Complex formation was endothermic with less than 1 kcal absorbed per mole of lipid or DNA charge. This enthalpy change was attributed to DNA-DNA mutual repulsion within the lamellar complex. The exception was DOTAP:DOPE-containing lipoplex for which the enthalpy of formation was exothermic, presumably because of DOPE amine group protonation. Experimental conditions, namely, direction and titration increment as well as concentration of titrant, which dictate the structure of resulting lipoplex (whether lamellar complex or DNA-coated vesicle), were found to affect the apparent thermodynamics of complex formation. The structure, in turn, influences the biological properties of the lipoplex. If the titration of lipid into DNA was carried out in large increments, the DeltaH was larger than when the injection increments were smaller, a finding that is consistent with increased vesicle disruption under large increments and which is expected theoretically. Cationic lipid-DNA binding was weak in high ionic strength solutions, however, the effective binding constant is within micromolar range because of macromolecular nature of the interaction. PMID- 12080143 TI - Metal ion-induced lateral aggregation of filamentous viruses fd and M13. AB - We report a detailed comparison between calculations of inter-filament interactions based on Monte-Carlo simulations and experimental features of lateral aggregation of bacteriophages fd and M13 induced by a number of divalent metal ions. The general findings are consistent with the polyelectrolyte nature of the virus filaments and confirm that the solution electrostatics account for most of the experimental features observed. One particularly interesting discovery is resolubilization for bundles of either fd or M13 viruses when the concentration of the bundle-inducing metal ion Mg(2+) or Ca(2+) is increased to large (>100 mM) values. In the range of Mg(2+) or Ca(2+) concentrations where large bundles of the virus filaments are formed, the optimal attractive interaction energy between the virus filaments is estimated to be on the order of 0.01 kT per net charge on the virus surface when a recent analytical prediction to the experimentally defined conditions of resolubilization is applied. We also observed qualitatively distinct behavior between the alkali-earth metal ions and the divalent transition metal ions in their action on the charged viruses. The understanding of metal ions-induced reversible aggregation based on solution electrostatics may lead to potential applications in molecular biology and medicine. PMID- 12080144 TI - VEGF receptor 2 and the adherens junction as a mechanical transducer in vascular endothelial cells. AB - Blood-flow interactions with the vascular endothelium represents a specialized example of mechanical regulation of cell function that has important physiological and pathophysiological cardiovascular consequences. Yet, the mechanisms of mechanostransduction are not understood fully. This study shows that shear stress induces a rapid induction as well as nuclear translocation of the vascular endothelial growth factor (VEGF) receptor 2 and promotes the binding of the VEGF receptor 2 and the adherens junction molecules, VE-cadherin and beta catenin, to the endothelial cytoskeleton. These changes are accompanied by the formation of a complex containing the VEGF receptor 2-VE-cadherin-beta-catenin. In endothelial cells lacking VE-cadherin, shear stress did not augment nuclear translocation of the VEGF receptor 2 and phosphorylation of Akt1 and P38 as well as transcriptional induction of a reporter gene regulated by a shear stress responsive promoter. These results suggest that VEGF receptor 2 and the adherens junction act as shear-stress cotransducers, mediating the transduction of shear stress signals into vascular endothelial cells. PMID- 12080145 TI - Positional cloning of the murine flavivirus resistance gene. AB - Inbred mouse strains exhibit significant differences in their susceptibility to viruses in the genus Flavivirus, which includes human pathogens such as yellow fever, Dengue, and West Nile virus. A single gene, designated Flv, confers this differential susceptibility and was mapped previously to a region of mouse chromosome 5. A positional cloning strategy was used to identify 22 genes from the Flv gene interval including 10 members of the 2'-5'-oligoadenylate synthetase gene family. One 2'-5'-oligoadenylate synthetase gene, Oas1b, was identified as Flv by correlation between genotype and phenotype in nine mouse strains. Susceptible mouse strains produce a protein lacking 30% of the C-terminal sequence as compared with the resistant counterpart because of the presence of a premature stop codon. The Oas1b gene differs from all the other murine Oas genes by a unique four-amino acid deletion in the P-loop located within the conserved RNA binding domain. Expression of the resistant allele of Oas1b in susceptible embryo fibroblasts resulted in partial inhibition of the replication of a flavivirus but not of an alpha togavirus. PMID- 12080146 TI - Human tumor-derived genomic DNA transduced into a recipient cell induces tumor specific immune responses ex vivo. AB - This article describes a DNA-based vaccination strategy evaluated ex vivo with human cells. The vaccine was prepared by transferring tumor-derived genomic DNA to PCI-13 cells, a highly immunogenic tumor cell line ("recipient cell"), which had been genetically modified to secrete IL-2 (PCI-13/IL-2). PCI-13 cells expressed class I MHC determinants (HLA-A2) shared with the tumor from which the DNA was obtained as well as allogeneic determinants. DNA from a gp100(+) melanoma cell line was transduced into gp100(-) PCI-13/IL-2 cells (PCI-13/IL-2/DNA). A T cell line specific for the gp100 epitope responded to PCI-13/IL-2/DNA cells by IFN-gamma-secretion measured in enzyme-linked immunospot assays. The T cell line also recognized the gp100 epitope presented by dendritic cells that ingested PCI 13/IL-2/DNA cells, which had been induced by UVB irradiation to undergo apoptosis. After up-take and processing of apoptotic PCI-13/IL-2/DNA cells, the dendritic cells primed normal peripheral blood lymphocytes to generate effector T cells specific for the tumor donating the DNA. The results indicate that tumor epitopes encoded in such DNA are expressed in recipient cells and can induce tumor-specific T cells. The findings support translation of this vaccination strategy to a phase I trial in patients with cancer. PMID- 12080147 TI - Ancestral polymorphism and adaptive evolution in the trichothecene mycotoxin gene cluster of phytopathogenic Fusarium. AB - Filamentous fungi within the Fusarium graminearum species complex (Fg complex) are the primary etiological agents of Fusarium head blight (scab) of wheat and barley. Scab is an economically devastating plant disease that greatly limits grain yield and quality. In addition, scabby grain is often contaminated with trichothecene mycotoxins that act as virulence factors on some hosts, and pose a serious threat to animal health and food safety. Strain-specific differences in trichothecene metabolite profiles (chemotypes) are not well correlated with the Fg complex phylogeny based on genealogical concordance at six single-copy nuclear genes. To examine the basis for this discord between species and toxin evolution, a 19-kb region of the trichothecene gene cluster was sequenced in 39 strains chosen to represent the global genetic diversity of species in the Fg complex and four related species of Fusarium. Phylogenetic analyses demonstrated that polymorphism within these virulence-associated genes is transspecific and appears to have been maintained by balancing selection acting on chemotype differences that originated in the ancestor of this important group of plant pathogens. Chemotype-specific differences in selective constraint and evidence of adaptive evolution within trichothecene genes are also reported. PMID- 12080150 TI - An "inverse satisfaction law"? Why don't older patients criticise health services? PMID- 12080152 TI - Equity in health. PMID- 12080153 TI - The concern for equity in health. PMID- 12080154 TI - The meaning and goals of equity in health. PMID- 12080155 TI - Development and validation of an instrument to measure perceived neighbourhood quality in Taiwan. AB - STUDY OBJECTIVE: s: Social epidemiologists have hypothesised that neighbourhood quality may exert an important contextual influence on mental and physical health. However, validated instruments do not exist for measuring neighbourhood quality in Taiwan. A self reported instrument to measure perceived neighbourhood quality in Taiwan was developed and tested. DESIGN: Community survey. SETTING: Southern Taiwan, including the metropolitan Kaohsiung area and eight surrounding communities, representing urban, suburban, and rural districts. PARTICIPANTS: A total of 1084 residents, aged 18 to 75, were surveyed during 1999 to 2000. MAIN RESULTS: Using factor analysis with varimax rotation, three subscales explained 54.8% of the variance in our 15 item Neighbourhood Quality Index: perceived social capital (Cronbach alpha=0.84), perceived security (alpha=0.78), and adequacy of services and facilities (alpha=0.67). Lower perceived neighbourhood social capital (odds ratio, OR, 1.26; 95% CI: 1.21 to 1.32), lower neighbourhood security (OR 1.37; 95% CI: 1.26 to 1.48), and inadequate neighbourhood services and facilities (OR 1.17; 95% CI: 1.06 to 1.28) were all related to higher residential dissatisfaction. CONCLUSIONS: A Neighbourhood Quality Index was developed for use in Taiwan with good internal consistency and test-retest reliability, as well as convergent validity. Future studies will examine the association between this index and measures of individual mental and physical health. PMID- 12080156 TI - Economic transition should come with a health warning: the case of Vietnam. AB - STUDY OBJECTIVE: s: To assess the affordability of health care to poor rural households in Vietnam under conditions of transition from a planned to a market economy and, in light of other transitional experience, inform policy on increasing access of the poor to affordable care of acceptable quality. DESIGN: Observational study by cross sectional socioeconomic survey, longitudinal healthcare seeking survey, and qualitative semi-structured interviews and focus group discussions; qualitative follow up over six years. SETTING: Four rural communes in north of Vietnam between 1992 and 1998. SURVEY PARTICIPANTS: 656 households (2995 people) selected by systematic random sampling. MAIN RESULTS: Compared with non-poor households, poor households had significantly lower average per capita rates of healthcare consultation and expenditure (p<0.01 in both cases). Poor households delayed and minimised healthcare seeking, especially of expensive hospital services. Two thirds of average healthcare spending by poor households was on relatively inexpensive but frequent acts of local ambulatory care. The poor restrained their healthcare seeking but not in proportion to income: for households reporting illness, the average proportion of income devoted to health care was 21.9% for the poor compared with 8.2% for the non-poor (p<0.01). To meet healthcare costs, many poor households reduced essential consumption, sold assets and incurred debt, threatening their future livelihood. CONCLUSIONS: In the short-term the poor need exemption from public sector user fees in both primary and hospital care. In the longer run the government budget and prepayment schemes should replace direct user charges in healthcare finance. Transitional economies like Vietnam should preserve the public health services built up under the planned economy. Market reforms that stimulate growth in the economy appear inappropriate to reform of social sectors. PMID- 12080157 TI - Effect of de-industrialisation on working conditions and self reported health in a sample of manufacturing workers. AB - STUDY OBJECTIVE: To explore the impact of de-industrialisation over a 20 year period on working conditions and health among sawmill workers, in the province of British Columbia (BC), Canada. DESIGN AND SETTING: This investigation is based on a sample of 3000 sawmill workers employed in 1979 (a year before the beginning de industrialisation) and interviewed in 1998. The sample was obtained by random selection from an already gathered cohort of approximately 28 000 BC sawmill workers. Change in working conditions from 1979 to 1998 are described. Self reported health status, in 1998, was used as a dependent variable in logistic regression after controlling for confounders. MAIN RESULTS: Downsizing in BC sawmills eliminated 60% of workers between 1979 and 1998. Working conditions in 1998 were better for those who left the sawmill industry and obtained re employment elsewhere. Workers who remained employed in restructuring sawmills were approximately 50% more likely to report poor health than those re-employed elsewhere. CONCLUSIONS: Working conditions and health status were better for workers who, under pressure of de-industrialisation, left the sawmill industry and obtained re-employment outside this sector. PMID- 12080159 TI - Outreach and improved access to specialist services for indigenous people in remote Australia: the requirements for sustainability. AB - STUDY OBJECTIVE: To examine the role of specialist outreach in supporting primary health care and overcoming the barriers to health care faced by the indigenous population in remote areas of Australia, and to examine issues affecting its sustainability. DESIGN: A process evaluation of a specialist outreach service, using health service utilisation data and interviews with health professionals and patients. SETTING: The Top End of Australia's Northern Territory, where Darwin is the capital city and the major base for hospital and specialist services. In the rural and remote areas outside Darwin there are many small, predominantly indigenous communities, which are greatly disadvantaged by a severe burden of disease and limited access to medical care. PARTICIPANTS: Seventeen remote health practitioners, five specialists undertaking outreach, five regional health administrators, and three patients from remote communities. MAIN RESULTS: The barriers faced by many remote indigenous people in accessing specialist and hospital care are substantial. Outreach delivery of specialist services has overcome some of the barriers relating to distance, communication, and cultural inappropriateness of services and has enabled an over fourfold increase in the number of consultations with people from remote communities. Key issues affecting sustainability include: an adequate specialist base; an unmet demand from primary care; integration with, accountability to and capacity building for a multidisciplinary framework centred in primary care; good communication; visits that are regular and predictable; funding and coordination that recognises responsibilities to both hospitals and the primary care sector; and regular evaluation. CONCLUSIONS: In a setting where there is a disadvantaged population with inadequate access to medical care, specialist outreach from a regional centre can provide a more equitable means of service delivery than hospital based services alone. A sustainable outreach service that is organised appropriately, responsive to local community needs, and has an adequate regional specialist base can effectively integrate with and support primary health care processes. Poorly planned and conducted outreach, however, can draw resources away and detract from primary health care. PMID- 12080158 TI - Equity in prevention and health care. AB - STUDY OBJECTIVE: There is an increasing body of evidence about socioeconomic inequality in preventive use, mostly for cancer screening. But as far as needs of prevention are unequally distributed, even equal use may not be fair. Moreover, prevention might be unequally used in the same way as health care in general. The objective of the paper is to assess inequity in prevention and to compare socioeconomic inequity in preventive medicine with that in health care. DESIGN: A cross sectional Health Interview Survey was carried out in 1997 by face to face interview and self administered questionnaire. Two types of health care utilisation were considered (contacts with GPs and with specialists) and four preventive care mostly delivered in a GP setting (flu vaccination, cholesterol screening) or in a specialty setting (mammography and pap smear). SETTING: Belgium. PARTICIPANTS: A representative sample of 7378 residents aged 25 years and over (participation rate: 61%). OUTCOME MEASURE: Socioeconomic inequity was measured by the HI(wvp) index, which is the difference between use inequality and needs inequality. Needs was computed as the expected use by the risk factors or target groups. MAIN RESULTS: There was significant inequity for all medical contacts and preventive medicine. Medical contacts showed inequity favouring the rich for specialist visits and inequity favouring the poor for contacts with GPs. Regarding preventive medicine, inequity was high and favoured the rich for mammography and cervical screening; inequity was lower for flu immunisation and cholesterol screening but still favoured the higher socioeconomic groups. In the general practice setting, inequity in prevention was higher than inequity in health care; in the specialty setting, inequity in prevention was not statistically different from inequity in health care, although it was higher than in the general practice setting. CONCLUSIONS: If inequity in preventive medicine is to be lowered, the role of the GP must be fostered and access to specialty medicine increased, especially for cancer screening. PMID- 12080160 TI - Smoke and mirrors: how Massachusetts diverted millions in tobacco tax revenues. AB - OBJECTIVE: This study examines the politics of appropriating Question 1 tobacco tax revenues in the first budget year after Massachusetts voters passed the ballot initiative in 1992. The initiative increased the tobacco tax on cigarettes by 25 cents per pack and on smokeless tobacco by 25% of the wholesale price. METHODS: Data were collected from newspapers, letters, memoranda, budgets, press releases, legislative floor debates, government documents, legislative journals, personal interviews, and tobacco industry documents that were downloaded from the Tobacco Archives internet site. RESULTS: During the first budget year, programmes mentioned by the initiative that were not exclusively tobacco related accounted for 27% of total Question 1 expenditures, while 50% of the revenues were allocated for programmes that were neither mentioned by the initiative nor provided any tobacco education, prevention, and cessation services. Only 23% of Question 1 funds were appropriated for programmes that provided exclusively tobacco education, prevention, and cessation services. Question 1 revenues were also used to supplant funding for pre-existing programmes, which was explicitly prohibited by the initiative. The first budget year became the template for Question 1 appropriations in subsequent fiscal years. CONCLUSION: Politics did not end after voters passed Question 1. Public health advocates lacked a strategy and budget plan to influence the appropriation of Question 1 funds after the passage of this ballot initiative. PMID- 12080161 TI - Genetics and social class. AB - OBJECTIVE: To assess claims that genes are a major determinant of social class. DESIGN: Using genetic epidemiological principles, five claims on the role of genes in determining social class are examined: (1) traits that run in families are usually inherited; (2) complex traits can be explained by alleles at a single gene locus; (3) complex traits are transmitted intact from one generation to the next; (4) natural selection explains social advantage. (5) Heritability estimates provide a valid estimate of the importance of genes in explaining complex human traits or behaviour. RESULTS: (1) Traits that run in families can result from environmental exposures that differ by social class. (2) The protein encoded by any single gene has too narrow a range of biological activity to explain traits as complex as social status. (3) Because alleles at different gene loci are transmitted independently, genetic inheritance cannot explain why offspring display the same complex traits as their parents. (4) The propagation of mutations that might result in a selective advantage takes much longer than the time for which any social class has achieved or maintained dominance. (5) Heritability measures are accurate only when environment is maintained constant. This is impossible in evaluating human traits. CONCLUSIONS: The roots of social class differences do not lie in our genes. Consequently, genetics cannot be used as a justification for maintaining a ruling class, limiting procreation among the poor, or minimising social support programmes. PMID- 12080162 TI - Mortality and bioclimatic discomfort in Emilia-Romagna, Italy. PMID- 12080163 TI - Trends in infant mortality inequalities in the Americas: 1955-1995. AB - STUDY OBJECTIVE: s: To describe overall and income related trends in infant mortality inequalities in the Region of the Americas from 1955 to 1995. DESIGN: Infant mortality rates (IMRs) were computed and their trends assessed by ordinary least squares. Overall trends in IMR inequalities among countries were analysed by comparing 10 year period IMRs, Gini coefficients, and Lorenz curves. Income related trends in IMR inequalities were assessed using 10 year period IMR ratios between the highest and the lowest quintiles of the per capita gross national product (GNP) distributions (adjusted for purchasing power). SETTING: Aggregated country data were used for all countries with over 200 thousand inhabitants (33 geopolitical units). The 10 year period midpoint IMR estimates used for the 1955 1995 time series were those published by the United Nations in 1997. MAIN RESULTS: IMRs decreased from 90.34 to 31.31 per 1000 live births between 1955 and 1995 at an average of 15.3 every 10 years. In contrast, Lorenz curves and Gini coefficients were similar for the five 10 year periods. After grouping by adjusted GNP distribution, a similar decreasing trend of IMR was observed in all groups. The rate ratio between the group at the lowest quintile and that at the highest quintile ranged from 4 to 5. The analysis of variance for repeated observations showed that there is a significant reduction in the IMR (F=130.18; p<0.01), that trends did not differ significantly among groups (F=1.16; p=0.32), and that they were approximately linear (F=155.83; p<0.01). CONCLUSIONS: Despite a sizable reduction in the infant mortality, whether or not income related, levels of IMR inequality among countries have remained almost constant between 1955 and 1995 in the Region of the Americas. Further analysis and focused interventions are needed to tackle the challenges of reducing these persistent mortality inequalities. PMID- 12080165 TI - Activity of African-American female teenagers in black organisations is associated with STD/HIV protective behaviours: a prospective analysis. PMID- 12080164 TI - Alcohol consumption and the incidence of type II diabetes. AB - BACKGROUND: This study examines the relation between alcohol and type II diabetes and the possible mediating effects of HDL-cholesterol and serum insulin. METHODS: Prospective study of 5221 men aged 40-59 years with no history of coronary heart disease, diabetes, or stroke drawn from general practices in 18 British towns. RESULTS: During the mean follow up of 16.8 years there were 198 incident cases of type II diabetes. Occasional drinkers were the reference group. A non-linear relation was seen between alcohol intake and age adjusted risk of diabetes, with risk lowest in light and moderate drinkers and highest in heavy drinkers (quadratic trend p=0.03). Further adjustment for body mass index decreased risk in heavy drinkers. After additional adjustment for physical activity, smoking, and (undiagnosed) pre-existing coronary heart disease, only moderate drinkers showed significantly lower risk than occasional drinkers (RR=0.66 95% CI 0.44 to 0.99). Alcohol intake was inversely associated with serum insulin and positively associated with HDL-cholesterol. Adjustment for these factors reduced the "protective" effect in moderate drinkers (adjusted RR=0.73 95% CI 0.48 to 1.10) but the quadratic trend remained significant (p=0.02). CONCLUSION: There is a non linear relation between alcohol intake and the risk of type II diabetes. Serum insulin and HDL-cholesterol explained a small amount (20%) of the reduction in risk of type II diabetes associated with moderate drinking. The adverse effect of heavy drinking seemed to be partially mediated through its effect on body weight. PMID- 12080166 TI - Appendicectomy, tonsillectomy, and inflammatory bowel disease: a case-control record linkage study. AB - STUDY OBJECTIVE: To determine whether appendicectomy and tonsillectomy are associated with ulcerative colitis (UC) or Crohn's disease (CD); and, if so, whether the associations are related to age at operation. DESIGN: Nested case control studies using a longitudinal database of linked hospital and death record abstracts. SETTING: Southern England. PATIENTS: Statistical records of people diagnosed with UC, CD, or a control condition admitted to hospitals in a defined area. MAIN RESULTS: Appendicectomy under the age of 20 years was associated with a significantly reduced subsequent risk of UC (relative risk =0.48, 95% confidence interval 0.30 to 0.73). The association appeared strongest for appendicectomy between 10 and 14 years of age (relative risk =0.29, 95% CI 0.09 to 0.68). Appendicectomy at the age of 20 years and over was associated with an increased subsequent risk of CD (relative risk =1.92, 95% CI 1.58 to 2.32), largely confined to those people whose CD was diagnosed within a year of appendicectomy. Appendicectomy under 20 years of age, undertaken five years or more before case or control conditions, was suggestively associated with a reduced risk of CD (relative risk =0.71, 95% CI 0.47 to 1.03). Prior tonsillectomy was not associated with any increase or decrease of risk of either UC or CD. CONCLUSIONS: Appendicectomy is associated with a reduced risk of UC; and the association is specific to young age groups when the population risk of appendicitis is itself highest. The increased risk of CD after appendicectomy, at short time intervals between the two, is probably attributable to the misdiagnosis of CD as appendicitis. PMID- 12080167 TI - Gender differences in clinical status at time of coronary revascularisation in Spain. AB - OBJECTIVE: To study gender differences in clinical status at the time of coronary revascularisation. DESIGN: Retrospective study of clinical records. Two stage stratified cluster sampling was used to select a nationally representative sample of patients receiving a coronary revascularisation procedure in 1997. SETTING: All of Spain. MAIN OUTCOME MEASURES: Odds ratios (OR) in men and women for different clinical and diagnostic variables related with coronary disease. A logistic regression model was developed to estimate the association between coronary symptoms and gender. RESULTS: In the univariate analysis the prevalence of the following risk factors for coronary heart disease was higher in women than in men: obesity (OR=1.8), hypertension (OR=2.9) and diabetes (OR=2.1). High surgical risk was also more prevalent among women (OR=2.6). In the logistic regression analysis women's risk of being symptomatic at the time of revascularisation was more than double that of men (OR=2.4). CONCLUSIONS: Women have more severe coronary symptoms at the time of coronary revascularisation than do men. These results suggest that women receive revascularisation at a more advanced stage of coronary disease. Further research is needed to clarify what social, cultural or biological factors may be implicated in the gender differences observed. PMID- 12080169 TI - DNA microarrays - a methodological breakthrough in genetics. PMID- 12080170 TI - A simplified AFLP method for fingerprinting of common wheat (Triticum aestivum L.) cultivars. AB - The simplified AFLP method was developed and evaluated for identification and genetic diversity studies of wheat cultivars. Selective primers exploited in AFLP assay based on a single cutting enzyme PstI ((PstI)AFLP) generated total of 111 robust fragments, including 67 (60%) monomorphic and 12 (11%) cultivar-specific markers. Average similarity between 15 cultivars was 0.650, and varied from 0.293 ('Hope' vs. 'Aurora') to 0.865 ('Norman' vs. 'Hornet'). Mean similarities within groups of winter wheat cultivars with and without 1BL/1RS chromosome were 0.713 and 0.685, respectively. A higher variation was found in the group of spring wheats: 0.677. The obtained results confirm the usefulness of the proposed modification of the AFLP technique for diversity studies and identification of common wheat cultivars. PMID- 12080171 TI - Genotypic and nutrition-dependent variation in water use efficiency and photosynthetic activity of leaves in winter wheat (Triticum aestivum L.). AB - Effects of high and reduced NPK nutrition on the genetic variation of components of water use efficiency at the leaf and whole-plant levels were examined in pot grown old and modern cultivars of winter wheat (Triticum aestivum L.). At the subsequent growth stages, the photosynthetic rate (A), transpiration rate (E), leaf area (LA) and gas exchange efficiency (A/E) were measured on fully developed 4th, 5th, penultimate and flag leaves. At the plant canopy level, the total amount of water transpired was recorded during the whole life cycle to determine the efficiency of water use in the vegetative (WUEveg) and grain (WUEgen) matter formation. Considerable genotypic differences were found for the characters studied. The limited NPK supply caused a decrease in LA, A and A/E, but contributed to an increase in WUEgen. Examined cultivars (C) did not interact with nutrition levels (N) for these characteristics. However, the position of leaves (L) and the C x L and N x L interactions significantly affected the variance in leaf photosynthetic characteristics. A and A/E were negatively correlated with LA, and the flag leaves were photosynthetically less active and less efficient per unit area than the lower leaves. The whole-plant components of WUE were found to be more genetically stable than the photosynthetic leaf characteristics. Some modern cultivars tended to form leaves of higher A and A/E than the older ones, and this corresponds with a more efficient use of water in grain formation (WUEgen) of the former. Stay-green duration of flag leaves and harvest index showed positive correlations with WUEgen. However, no close associations were noticed between WUE components and stress tolerance, and the modern cultivars were usually less tolerant to NPK shortage. PMID- 12080172 TI - Resistance of Polish lines and hybrids of watermelon [Citrullus lanatus (Thunb.) Matsum et Nakai] to Fusarium oxysporum at the seedling stage. AB - Watermelon is a species cultivated in the hot climate or in the greenhouse. Since recently it has also started to be grown in the open in the Polish climate. This species is frequently at risk of Fusarium oxysporum infection. Between 1996 and 1997 ten inbred lines and nine hybrids of Polish origin were tested for resistance to this pathogen. The test was conducted with the use of four isolates of F. oxysporum: three from Polish infected plants (formae speciales not determined), while the fourth from U.K. (F. oxysporum f. sp. niveum). In the three series of tests the control plants were Pannonia F(1) and Sugar Baby. No inbred line or hybrid was found to be highly resistant to the pathogen. However, it was possible to identify four lines and five hybrids showing a higher level of resistance as compared with the control. The level of hybrid resistance was determined by comparison with the parental genotypes. PMID- 12080173 TI - A simple test for detection of linkage. AB - The paper presents a proposition for detection of linkage of genes responsible for metrical traits. Taking into account the expected means for early generations (F(1), F(2), F(3)) and a population of homozygous lines (in this case doubled haploid lines, DH derived from a cross between two homozygous parents) as well as estimators of genetic parameters m, [d], [i], [h] and [l], the expected values for these parameters in the presence of linkage have been formulated. It was found that when there is no linkage, the expression F(1) - 6F(2) + 8F(3) - 3DH(mean) is equal to zero. Thus, an experiment covering DH lines and F(1), F(2), F(3) hybrids makes it possible to obtain, beside information of interest, also information on presence or absence of linkage. PMID- 12080174 TI - Novel linkage mapping approach using DNA pooling in human and animal genetics. I. Detection of complex disease loci. AB - DNA pooling is a potential methodology for genetic loci with small effect contributing to complex diseases and quantitative traits. This is accomplished by the rapid preliminary screening of the genome for the allelic association with the most common class of polymorphic short tandem repeat markers. The methodology assumes as a common founder for the linked disease locus of interest and searches for a region of a chromosome shared between affected individuals. The general theory of DNA pooling basically relies on the observed differences in the allelic distribution between pools from affected and unaffected individuals, including a reduction in the number of alleles in the affected pool, which indicate the sharing of a chromosomal region. The power of statistic for associated linkage mapping can be determined using two recently developed strategies, firstly, by measuring the differences of allelic image patterns produced by two DNA pools of extreme character and secondly, by measuring total allele content differences by comparing between two pools containing large numbers of DNA samples. These strategies have effectively been utilized to identify the shared chromosomal regions for linkage studies and to investigate the candidate disease loci for fine structure gene mapping using allelic association. This paper outlines the utilization of DNA pooling as a potential tool to locate the complex disease loci, statistical methods for accurate estimates of allelic frequencies from DNA pools, its advantages, drawbacks and significance in associate linkage mapping using pooled DNA samples. PMID- 12080175 TI - Prediction of additive genetic effects for the QTL-cluster on the basis of data on surrounding markers in outbred populations. AB - This paper described a method for predicting additive effects of a cluster of tightly linked QTLs for outbred populations of animals in the situation where the QTLs are located on a chromosome segment surrounded by multiple linked DNA markers. We present a mixed model method for best linear unbiased prediction (conditional to the marker data) of the additive effects of the QTL-cluster and of the remaining QTLs unlinked to the marker linkage group. This method takes into consideration the identity-by-descent proportion (IBDP) for the particular chromosomal segment, in contrast to some other methods which use IBD probabilities at one specific location. In this method, fully informative data on different flanking markers is used to calculate the values of the expectations of the IBDPs (EIBDPs) between gametes for animals to be evaluated. Then the expected values are used as the elements of the gametic relationship matrix required in the best linear unbiased prediction. Giving a small numerical example, we illustrate how the present method can be used for the prediction of the QTL cluster effects and for genetic evaluation of animals in outbred populations. A computational strategy is discussed on the basis of the calculation of the EIBDPs and the inverted gametic relationship matrix in complex pedigrees. PMID- 12080176 TI - National evaluation for calving ease, gestation length and birth weight by linear and threshold model methodologies. AB - Data included 393,097 calving ease, 129,520 gestation length, and 412,484 birth weight records on 412,484 Gelbvieh cattle. Additionally, pedigrees were available on 72,123 animals. Included in the models were effects of sex and age of dam, treated as fixed, as well as direct, maternal genetic and permanent environmental effects and effects of contemporary group (herd-year-season), treated as random. In all analyses, birth weight and gestation length were treated as continuous traits. Calving ease (CE) was treated either as a continuous trait in a mixed linear model (LM), or as a categorical trait in linear-threshold models (LTM). Solutions in TM obtained by empirical Bayes (TMEB) and Monte Carlo (TMMC) methodologies were compared with those by LM. Due to the computational cost, only 10,000 samples were obtained for TMMC. For calving ease, correlations between LM and TMEB were 0.86 and 0.78 for direct and maternal genetic effects, respectively. The same correlations but between TMEB and TMMC were 1.00 and 0.98, respectively. The correlations between LM and TMMC were 0.85 and 0.75, respectively. The correlations for the linear traits were above.97 between LM and TMEB but as low as 0.91 between LM and TMMC, suggesting insufficient convergence of TMMC. Computing time required was about 2 hrs, 5 hrs, and 6 days for LM, TMEB and TMMC, respectively, and memory requirements were 169, 171, and 445 megabytes, respectively. Bayesian implementation of threshold model is simple, can be extended to multiple categorical traits, and allows easy calculation of accuracies; however, computing time is prohibitively long for large models. PMID- 12080177 TI - Microsatellite polymorphism in locus OMHC1 (MHC Class I) in Polish Heath Sheep and Polish Lowland Sheep (Zelazna variety). AB - Our study aimed at comparative analysis of microsatellite polymorphism in locus OMHC1 (MHC Class I) in Polish Heath Sheep and Polish Lowland Sheep (Zelazna variety). The study was conducted on 100 ewes of each breed. We identified 13 alleles of the gene in Polish Heath Sheep and 9 in Polish Lowland Sheep. We found marked differences in frequency of OMHC1 alleles between both breeds. The heterozygosity coefficient and PIC, amounting to 0.79 and 0.77 for Polish Heath Sheep, and 0.82 and 0.80 for Polish Lowland Sheep, respectively, suggest considerable variability in both breeds. Additionally, the values of both coefficients indicate that OMHC1 locus can be used as a genetic marker. PMID- 12080178 TI - Clinical features, treatment and genetic background of Treacher Collins syndrome. AB - Treacher Collins syndrome (TCS) is an autosomal dominant disorder of craniofacial development. The major features of the disease include midface hypoplasia, micrognathia, microtia, conductive hearing loss and cleft palate. Current procedures of surgical treatment of TCS are discussed and novel findings concerning the genetic background of TCS are described. The TCS locus has been mapped to chromosome 5q31.3-32. The TCOF1 gene contains 26 exons and encodes a 1411 amino acid protein named treacle. In the TCOF1 gene 51 mutations have been identified. Most of these mutations are insertions or deletions, which result in an introduction of a premature termination codon into the reading frame. Mutational spectra support the hypothesis that TCS results from haploinsufficiency of treacle. PMID- 12080179 TI - Novel competitive PCR methods for quantitation of T-cell receptor delta (TCRD) gene rearrangements. AB - Since the invention of the polymerase chain reaction (PCR) several quantitative PCR-based approaches have been described. Recently, the real-time PCR method became a standard in quantitative PCR, although high costs of the necessary equipment and reagents make it unaffordable for many laboratories. In this paper we describe two novel competitive PCR techniques, which were used to determine the frequency of T-cell receptor delta gene (TCRD) rearrangements in peripheral blood leukocytes. In the reference gene competitive PCR (rgc-PCR) the rearranged TCRD gene competes with the reference gene (RAG1) for common reagents (dNTPs and Taq polymerase). The intensity ratio of amplification products, TCRD/RAG1, corresponds to the portion of cells containing a rearrangement. A series of reactions was performed, in which RAG1 primers were added to the PCR after different numbers of cycles. On the basis of the number of cycles needed to obtain equal band intensity, the frequency of cells containing a rearrangement was calculated. In the common primer competitive PCR (cpc-PCR), two gene rearrangements, Vdelta1-Jdelta1 and Vdelta2-Jdelta1, compete for the common Jdelta1 primer. The competing genes are amplified from the same genomic DNA template; therefore unlike in the method using the internal competitor, the results are not affected by the quantity or quality of the analysed sample. We showed that the rgc-PCR and cpc-PCR are reliable and give reproducible results. The methods do not require any expensive equipment or reagents, and can be used to determine the frequency of gene rearrangements. PMID- 12080180 TI - The KVLQT1 gene is not a common target for mutations in patients with various heart pathologies. AB - The long QT syndrome (LQTS) is a disorder of ventricular repolarization that exposes affected individuals to cardiac arrhythmias and sudden death. The first gene for LQTS has been mapped to chromosome 11 p.15.5 by genome-wide linkage analysis. This gene, originally named KVLQT1 (and later KCNQ1), is a novel potassium channel gene. Mutations in the human KVLQT1 gene, encoding the alpha subunit of the KVLQT1 channel, cause the long QT syndrome. In this work, we analysed the sequence of six KVLQT1 exons in patients with various heart pathologies. We describe 6 different mSSCP patterns with no disease-related SSCP conformers in any sample. Direct sequencing of exons 2 to 7 confirmed the absence of mutations. This suggests that the analysed region of the KVLQT1 gene is not commonly involved in pathogenesis of the long QT syndrome. PMID- 12080181 TI - Variability of the human mitochondrial DNA control region sequences in the Lithuanian population. AB - The Lithuanians and Latvians are the only two Baltic cultures that survived until today. Since the Neolithic period the native inhabitants of the present-day Lithuanian territory have not been replaced by any other ethnic group. Therefore the genetic characterization of the present-day Lithuanians may shed some light on the early history of the Balts. We have analysed 120 DNA samples from two Lithuanian ethnolinguistic groups (Aukstaiciai and Zemaiciai) by direct sequencing of the first hypervariable segment (HVI) of the control region of mitochondrial DNA (mtDNA) and restriction enzyme digestion for polymorphic site 00073. On the basis of specific nucleotide substitutions the obtained sequences were classified to mtDNA haplogroups. This revealed the presence of almost all European haplogroups (except X) in the Lithuanian sample, including those that expanded through Europe in the Palaeolithic and those whose expansion occurred during the Neolithic. Molecular diversity indices (gene diversity 0.97, nucleotide diversity 0.012 and mean number of pairwise differences 4.5) were within the range usually reported in European populations. No significant differences between Aukstaiciai and Zemaiciai subgroups were found, but some slight differences need further investigation. PMID- 12080182 TI - [alpha]-Secretase ADAM10 as well as [alpha]APPs is reduced in platelets and CSF of Alzheimer disease patients. AB - BACKGROUND: Members of membrane-bound disintegrin metalloproteinases (ADAMs) were shown to be capable of cleaving amyloid precursor protein (APP) at the alpha cleavage site in different cell systems. One of the candidate alpha-secretases identified in this family is ADAM10. The present study addresses the following major questions: 1) Are the levels of an alpha-secretase candidate (i.e., ADAM10) reduced in accessible cells of Alzheimer Disease (AD) patients? 2) Are ADAM10 levels in the peripheral cells of AD patients related to a concomitant decrease in alpha APPs? MATERIALS AND METHODS: Western Blot analysis of ADAM10 is performed on platelet homogenates from 33 sporadic AD patients and on 26 age matched control subjects. Moreover, the levels of alpha-secretase metabolite (alpha APPs) are tested both in platelets and cerebrospinal fluid (CSF) of the same pool of subjects by means of Western blot with a specific antibody. RESULTS: A significant decrease of platelet ADAM10 levels is observed in patients affected by probable AD when compared to control subjects and this is paralleled by a reduced level of alpha APPs released from platelets. Moreover, in the same pool of AD patients, alpha APPs levels were reduced concomitantly in CSF. CONCLUSIONS: ADAM10 is expressed in platelets. A reduced level of ADAM10 is observed in platelets obtained from AD patients compared to age-matched controls. Further, in the same pool of AD patients, a qualitatively and quantitatively similar decrease in alpha APPs is present both in thrombin-activated platelets and CSF, thus suggesting that alterations of APP processing might occur both in the neuronal compartment and peripheral cells. PMID- 12080183 TI - A novel natural product compound enhances cAMP-regulated chloride conductance of cells expressing CFTR[delta]F508. AB - BACKGROUND: Cystic fibrosis (CF) results from mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which encodes a chloride channel localized at the plasma membrane of diverse epithelia. The most common mutation leading to CF, Delta F508, occurs in the first nucleotide-binding domain (NBD1) of CFTR. The Delta F508 mutation disrupts protein processing, leading to a decreased level of mutant channels at the plasma membrane and reduced transepithelial chloride permeability. Partial correction of the Delta F508 molecular defect in vitro is achieved by incubation of cells with several classes of chemical chaperones, indicating that further investigation of novel small molecules is warranted as a means for producing new therapies for CF. MATERIALS AND METHODS: The yeast two-hybrid assay was used to study the effect of CF causing mutations on the ability of NBD1 to self-associate and form dimers. A yeast strain demonstrating defective growth as a result of impaired NBD1 dimerization due to Delta F508 was used as a drug discovery bioassay for the identification of plant natural product compounds restoring mutant NBD1 interaction. Active compounds were purified and the chemical structures determined. The purified compounds were tested in epithelial cells expressing CFTR Delta F508 and the resulting effect on transepithelial chloride permeability was assessed using short-circuit chloride current measurements. RESULTS: Wild type NBD1 of CFTR forms homodimers in a yeast two-hybrid assay. CF-causing mutations within NBD1 that result in defective processing of CFTR (Delta F508, Delta I507, and S549R) disrupted NBD1 interaction in yeast. In contrast, a CF causing mutation that does not impair CFTR processing (G551D) had no effect on NBD1 dimerization. Using the yeast-based assay, we identified a novel limonoid compound (TS3) that corrected the Delta F508 NBD1 dimerization defect in yeast and also increased the chloride permeability of Fisher Rat Thyroid (FRT) cells stably expressing CFTR Delta F508. CONCLUSION: The establishment of a phenotype for the Delta F508 mutation in the yeast two-hybrid system yielded a simple assay for the identification of small molecules that interact with the mutant NBD1 and restore dimerization. The natural product compound identified using the system (TS3) was found to increase chloride conductance in epithelial cells to an extent comparable to genistein, a known CFTR activator. The yeast system will thus be useful for further identification of compounds with potential for CF drug therapy. PMID- 12080184 TI - The alpha 2-adrenergic receptor gene and body fat content and distribution: the HERITAGE Family Study. AB - BACKGROUND: Among adrenergic receptor subtypes that regulate lipid mobilization, the alpha2-adrenergic receptor is involved in the inhibition of fatty acid mobilization from adipose tissue. A C-1291G polymorphism is located in the alpha2 adrenergic receptor gene (ADRA2A) but no association with body fat accumulation has been reported yet. MATERIALS AND METHODS: Body mass index (BMI), fat mass (FAT), percentage body fat (%FAT), trunk-to-extremity skinfold ratio (TER), sum of eight skinfolds (SF8), and abdominal subcutaneous (ASF), visceral (AVF), and total (ATF) fat areas assessed by CT scan have been measured in adult sedentary white (n = 503) and black (n = 276) subjects participating in the HERITAGE Family Study. Association between the C-1291G polymorphism and each phenotype was tested separately in men and women of each race using ANCOVA with the effects of age as covariate in addition to the effects of BMI for TER and of FAT for AVF, ASF, and ATF. RESULTS: The allele frequencies of the ADRA2A C-1291G polymorphism differed between races. No association was observed in white subjects, except for a moderate effect of the polymorphism accounting for less than 1% of the variance in AVF and ATF in women. In black subjects, however, the G-1291 allele was found to be associated with an increase of TER in men (3.8% of variance accounted for by the polymorphism), while in black women it was associated with a decrease in TER (2.9%) and in AVF (2.5%). CONCLUSION: These results suggest a role for the ADRA2A gene in determining the propensity to store fat in the abdominal area, independently of total body fatness. PMID- 12080185 TI - Nonredundant antioxidant defense by multiple two-cysteine peroxiredoxins in human prostate cancer cells. AB - BACKGROUND: Peroxiredoxins (Prxs) are antioxidant enzymes expressed by most free living organisms, often in multiple isoforms. Because mammalian Prxs have not been experimentally deleted or inhibited, it is not known how much they contribute to antioxidant defense, nor whether the multiple isoforms afford redundant or additive protection. MATERIALS AND METHODS: Expression of the four members of the 2-Cys family of human Prxs was tested in human tumor cell lines. Monospecific antibodies were developed and used to monitor the extent and specificity of inhibition of expression of each isoform in prostate cancer cells stably transfected with antisense constructs. RESULTS: Seventeen tumor lines transcribed genes for all four human Prxs. Prostate cancer cells coexpressed each isoform at the protein level. Stable transfection with antisense allowed partial, selective suppression of Prx 1, 2, 3, or 4. Prostate cancer cells were rendered more sensitive to hydrogen peroxide or an organic hydroperoxide when Prx 1, 2, or 3 but not 4 was partially suppressed, bringing them into the range of sensitivity of mouse cells. The effect of partially suppressing a single Prx was comparable to that of depleting glutathione. In contrast, sensitization to adriamycin, an antitumor agent with a redox-active quinone, followed the partial suppression of Prxs 1, 2, or 4 but not 3. Individual suppression of Prxs 1-4 had no effect on sensitivity of the cells to reactive nitrogen intermediates, tumor necrosis factor (TNF), paclitaxel (Taxol), or etoposide. CONCLUSIONS: The 2-Cys Prxs act in a mutually nonredundant and sometimes stress-specific fashion to protect human cells from oxidant injury. The substantial resistance of human cells to hydroperoxides may result in part from the additive action of multiple Prxs. PMID- 12080203 TI - States' spending for public welfare and their suicide rates, 1960 to 1995: what is the problem? AB - Drawing on Durkheim's theory of social integration, this discussion reports on findings from a pooled time-series analysis of states' spending for public welfare and their suicide rates, controlling for states' divorce rates, population change rates, population density, unemployment rates, sex ratio, and racial composition. The analysis spans a 35-year period, 1960 to 1995, at six different data points: 1960, 1970, 1980, 1985, 1990, and 1995. The major hypothesis was that states' suicide rates would increase with decreases in per capita spending for public welfare, controlling for the variables listed above in three different models and using OLS to analyze the data. In the basic model, states' spending for public welfare showed no relationship to states' suicide rates; in the second model that controlled for data year and in the third model that controlled for both data year and state, its relationship was significant, but in a negative direction. Suicide rates increased in states that reduced their per capita expenditures for public welfare during the observational period. Of all the variables, the influence of divorce on suicide was the most persistent and pronounced, followed by the percentage of whites in states' populations. Whether the findings reflect an increase in the unendurable psychological pain associated with suicide, or the weakening of ties that bind individuals to each other and to the larger society (as measured by states' divorce rates and per capita expenditures for public welfare), or the vulnerabilities associated with race, states can help counter suicide trends and such negative influences as divorce as evidenced by states that spend more for public welfare and have lower suicide rates. Given that clinicians work with people experiencing the unendurable psychological pain associated with suicide, the findings from these analyses have relevance for their practice. PMID- 12080186 TI - Noninvasive dynamic fluorescence imaging of human melanomas reveals that targeted inhibition of bFGF or FGFR-1 in melanoma cells blocks tumor growth by apoptosis. AB - BACKGROUND: Two prominent biological features of the advanced stages of human melanoma are their high degree of vascularity and high-level expression of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor-1 (FGFR-1). Given these characteristics, human melanoma serves as an ideal model to address an important question regarding the efficacy of angiogenesis-based cancer therapy. To induce tumor growth arrest and regression, does it suffice to block expression of bFGF and/or FGFR-1 in only the melanoma cells, or is it essential to inhibit expression of bFGF and/or FGFR-1 in both the melanoma cells and the melanoma cell-interspersing vasculature? MATERIALS AND METHODS: Primary and metastatic human melanomas, grown as subcutaneous tumors in nude mice, were injected twice a week with vector constructs containing the human tyrosinase promoter and antisense- oriented human bFGF or FGFR-1 cDNA. On alternating days, the bFGF and FGFR-1 antisense-targeted tumors received injections of cyanine fluorochrome-conjugated antibodies to a human melanoma and mouse blood vessel marker. Noninvasive, dynamic fluorescence imaging was used to document the cellular events that took place inside the tumors as the result of blocking expression of bFGF or FGFR-1 in the melanoma cells. RESULTS: In vivo, ex vivo, and in vitro fluorescence imaging of the bFGF and FGFR-1 antisense-targeted tumors demonstrated that inhibiting bFGF and FGFR-1 signaling in only the melanoma cells suffices to inhibit tumor growth due to massive induction of melanoma cell apoptosis. CONCLUSIONS: The investigations presented in this study document that inhibiting expression of bFGF or FGFR-1 in only the melanoma cells is as effective in blocking tumor growth as simultaneously inhibiting bFGF or FGFR-1 synthesis in the melanoma cells and the melanoma cell-interspersing vasculature. Furthermore, blocking expression of bFGF or FGFR-1 in the melanoma cells did not lead to activation or increased production of another angiogenic molecule, suggesting the absence of a "salvage pathway" that can circumvent or rescue the blockage of bFGF/FGFR-1 in the melanoma cells. PMID- 12080204 TI - Commentary on states' spending. PMID- 12080206 TI - Recognizing the anxious face of depression. AB - "Anxious depression" is used variably both by researchers and clinicians to describe admixtures of anxiety and depressive symptoms. The authors sought to determine the best model for conceptualizing anxious depression by studying a sample of depressed patients referred to a tertiary referral unit. Anxiety and depression were assessed using a comprehensive set of mixed symptoms that were subsequently refined to provide separate anxiety and depressive factors, and patients were trichotomized into groups of low, medium, and high anxiety on the basis of their total anxiety factor scores. Associations between the constructs of anxiety and depression in different depressive subgroups were explored, and the severity of depressive symptoms and other clinical variables across the three anxiety groupings was assessed. Depression variables were not linearly associated in a consistent pattern with anxiety-defined groups, arguing against a simple interdependence model driven by a higher-order variable such as depression severity. By contrast, the state anxiety categories were linked strongly with lifetime anxiety disorder prevalence, with some associations linear and with others evidencing a trend break association. The authors found support for a model of anxious depression, whereby anxiety both predisposes to nonmelancholic depression and contributes to its presentation by shaping its clinical features. Such a model and its definition assist in clarifying the cause of anxious depression and its treatment. PMID- 12080207 TI - The mental health of war-wounded refugees: an 8-year follow-up. AB - The complex nature of recent wars and armed conflicts has forced many war-injured persons into exile. To investigate their long-term mental health, three instruments for assessing mental health (HSCL-25, PTSS-10, and a Well-Being scale) were presented to 44 war-wounded refugees from nine different countries 8 years after arrival in Sweden. The prevalence of psychiatric symptoms was high and corresponds to findings in previous studies of refugee patient populations. A lower level of mental health was associated with higher education, unemployment, and poor physical health. The findings suggest a high psychiatric morbidity and a need for psychiatric interventions in this refugee group. Methodological issues to be considered in research on sequels of war traumas are discussed. PMID- 12080208 TI - Severity of reported childhood sexual abuse and its relationship to severity of borderline psychopathology and psychosocial impairment among borderline inpatients. AB - This study has two purposes. The first purpose is to describe the severity of sexual abuse reported by a well-defined sample of borderline inpatients. The second purpose is to determine the relationship between the severity of reported childhood sexual abuse, other forms of childhood abuse, and childhood neglect and the severity of borderline symptoms and psychosocial impairment. Two semistructured interviews of demonstrated reliability were used to assess the severity of adverse childhood experiences reported by 290 borderline inpatients. It was found that more than 50% of sexually abused borderline patients reported being abused both in childhood and in adolescence, on at least a weekly basis, for a minimum of 1 year, by a parent or other person well known to the patient, and by two or more perpetrators. More than 50% also reported that their abuse involved at least one form of penetration and the use of force or violence. Using multiple regression modeling and controlling for age, gender, and race, it was found that the severity of reported childhood sexual abuse was significantly related to the severity of symptoms in all four core sectors of borderline psychopathology (affect, cognition, impulsivity, and disturbed interpersonal relationships), the overall severity of borderline personality disorder, and the overall severity of psychosocial impairment. It was also found that the severity of childhood neglect was significantly related to five of the 10 factors studied, including the overall severity of borderline personality disorder, and that the severity of other forms of childhood abuse was significantly related to two of these factors, including the severity of psychosocial impairment. Taken together, the results of this study suggest that the majority of sexually abused borderline inpatients may have been severely abused. They also suggest that the severity of childhood sexual abuse, other forms of childhood abuse, and childhood neglect may all play a role in the symptomatic severity and psychosocial impairment characteristic of borderline personality disorder. PMID- 12080209 TI - Is there a differential impact of fatigue and physical disability on quality of life in multiple sclerosis? AB - To investigate the quantitative impact of fatigue on health-related quality of life (HRQoL) in multiple sclerosis (MS) and to determine whether fatigue was related to HRQoL independently from bodily disability, data on HRQoL were ascertained for 87 patients with definite MS by using the SF-36. HRQoL scores and subscores were related to the basic MS disability score (EDSS) and further MS parameters, and to fatigue, which was assessed by using different fatigue scales. Factors related to predominantly physical but not mental HRQoL aspects were identified as related to EDSS, duration of disease, and age. Different fatigue scores did impact significantly on both physical and especially mental HRQoL. The influence of fatigue on physical HRQoL was independent from EDSS. Fatigue experience reduces HRQoL markedly and independently from EDSS. Therefore, fatigue assessment provides additional information to disability-derived scales such as the EDSS with relevant implications for therapeutic decisions. PMID- 12080211 TI - Historical note. The ability of mentally ill persons to live in the community, 1880. PMID- 12080210 TI - Local perceptions of the mental health effects of the Uganda acquired immunodeficiency syndrome epidemic. AB - Despite much attention in developed countries, little is known about the relationship between mental health problems and the human immunodeficiency virus (HIV) in Africa. The objectives of the current study were a) to investigate how people in an African community severely affected by HIV view the mental health effects of the epidemic and b) to use these data to investigate the local construct validity of the Western concepts of depression and posttraumatic stress disorder. Ethnographic methods-free listing and key-informant interviews-were used among participants from the Rakai and Masaka districts of southwest Uganda. Participants described two independent depression-like syndromes (Yo'kwekyawa and Okwekubaziga) resulting from the HIV epidemic. No syndromes similar to posttraumatic stress disorder were detected. We conclude that local people recognize depression syndromes and consider them pertinent consequences of the HIV epidemic. PMID- 12080212 TI - Factor structure of the Multnomah Community Ability Scale. AB - More and more, Intensive Community Treatment programs in Canada use questionnaires to evaluate the health and social functioning of individuals suffering from a severe mental illness. However, seldom are these tools subject to psychometric analyses to establish their validity on independent samples. This article presents the results of the validation of the French version of the Multnomah Community Ability Scale. Confirmatory factor analyses were carried out to assess the factorial structure. The factor structure, four dimensions with three items, emerging from a first sample was replicated with data from a different sample of clients. Moreover, these four dimensions respect the initial factor solution of the Multnomah Community Ability Scale, which are a) interference with functioning, b) adjustment to living, c) social competence, and d) behavior problems. The study shows the structural validity of this brief questionnaire, which could be useful both for clinical and research settings to evaluate the effectiveness of interventions. PMID- 12080213 TI - Evidence supporting a cognitive model of depression in Parkinson's disease. AB - The objective of this study was to explore the possibility that depression in Parkinson disease (PD) arises from an illness-related vulnerability to negative emotional stimuli. The Color-Word and Emotional Stroop tasks were performed by 27 nondepressed patients with PD and 27 healthy volunteers. Patients with PD showed greater interference to the sad words of the Emotional Stroop than did control patients, despite similar performances to the controls on the neutral and Color Word Stroop tasks. Nondepressed patients with PD may have attentional mechanisms that are vulnerable to negative stimuli. This vulnerability might arise out of a common involvement of anterior cingulate regions in both selective attention and mood regulation, and could in turn predispose to cognitive distortions that may lead to depression. PMID- 12080214 TI - Current progress in schizophrenia research. Search for genes of schizophrenia: back to defining valid phenes. PMID- 12080215 TI - Associations between juvenile and adult forms of separation anxiety disorder: a study of adult volunteers with histories of school refusal. PMID- 12080217 TI - Proposal for a new grading system for chronic pancreatitis: the ABC system. PMID- 12080218 TI - The alkaline tide phenomenon. AB - The parietal cell is capable of secreting high concentrations of hydrochloric acid into the lumen of the stomach. The apical membrane of this cell contains K+H+ ATPase, which is responsible for proton transport into the lumen. Potassium and chloride channels are also present. The basolateral membrane of the parietal cell possesses transporters that maintain intracellular homeostasis. Specifically, large amounts of bicarbonate that are generated by carbonic anhydrase must be removed from the cell to prevent alkalinization. Efflux of bicarbonate into the blood after acid secretion can be detected and is known as the alkaline tide. Determination of the alkaline tide has been used to measure acid secretion. In this review, we summarize the published data. PMID- 12080219 TI - Familial Crohn's disease in single or multiple first-degree relatives. AB - The strongest currently recognized risk factor for Crohn's disease (CD) development is having a relative with the disease. In this study, 1,000 patients with CD, seen directly by the investigator during a period of more than 20 years, were consecutively evaluated for a family history of CD. There were 140 patients who reported a relative with CD. Of these, all 87 first-degree relatives were confirmed to have CD (36 men and 51 women). Siblings, particularly women, were most commonly affected in comparison with parents or children. There were 65 with a single affected first-degree relative and 22 with multiple affected first degree relatives. In this series, the tendency to have a female sibling with CD was further increased in those with multiple affected first-degree relatives. Although the age of diagnosis of children was less than the age of diagnosis in the respective parents, there were almost identical numbers of mother-child and father-child pairs. These findings in a population with prolonged follow-up by the same physician may reflect a bias toward case ascertainment rather than the previously reported phenomena in recruited populations of genetic anticipation and genomic imprinting. Patients with multiple first-degree relatives were older with more extensive disease in both ileal and colonic sites compared with those with only a single first-degree relative and more frequent colonic disease. Future studies are needed to further explore the role of both genetic and environmental factors in the pathogenesis and phenotypic expression of familial CD. PMID- 12080220 TI - The economic impact of irritable bowel syndrome in a managed care setting. AB - GOALS: To compare the healthcare resource consumption of patients who have irritable bowel syndrome (IBS) with an age-, gender-, and comorbidity-matched cohort of patients without IBS from a managed care perspective. STUDY: Retrospective cohort analysis. Data were obtained electronically through the Henry Ford Health System corporate data warehouse. Patients with IBS were eligible if they had at least one primary diagnosis of irritable colon (ICD-9-CM 564.1). The control cohort was identified from an age, gender, and comorbidity population-matched (5:1) sample. The index date for the IBS and control cohorts was defined as the first initial diagnosis and first clinical encounter in 1998, respectively. The charges per patient by resource use type were collected for the 12 months before and after the index date. RESULTS: A total of 501 patients with IBS and 2505 controls fulfilled the inclusion and exclusion criteria (70% female, 58% between 40 and 65 years). The IBS cohort, before the IBS diagnosis, had significantly higher total charges (p < 0.001), drug charges (p < 0.001), and outpatient charges (p < 0.001) than the control cohort. During the postindex (postdiagnosis period), the patients with IBS had higher total charges (p < 0.006), outpatient charges (p < 0.022), and drug charges (p < 0.001) than the control population. The control cohort had higher procedure charges (p < 0.001) during both periods. CONCLUSIONS: Patients with an IBS diagnosis represent a substantial cost to managed care before and after the diagnosis of IBS. Costs associated with these patients result mainly from non-IBS conditions. Further research is warranted to identify these patients earlier, and to prevent the economic burden associated with them. PMID- 12080221 TI - Orocecal transit time in patients with functional dyspepsia. AB - BACKGROUND: The importance of intestinal dysmotility in functional dyspepsia is a controversial issue. GOALS: To investigate the orocecal transit time in patients with functional dyspepsia, as well as a possible association between intestinal transit and the presence of anxiety or Helicobacter pylori infection in these patients. STUDY: The participants in this study were 23 patients with dysmotility like functional dyspepsia and 24 control subjects. Orocecal transit time was measured by the lactulose hydrogen breath test. The presence of anxiety was assessed by the Hospital Anxiety and Depression (HAD) scale. RESULTS: No significant difference in orocecal transit times was found between patients (median, 55 minutes; 95% confidence interval [CI], 40-60 minutes) and control subjects (median, 50 minutes; 95% CI, 40-60 minutes; p = 1). In the assessment, 15 patients (73%) scored for anxiety on the HAD scale, and 15 patients (65%) had positive test results for H. pylori. There was no significant difference in orocecal transit times between the patients with (median, 55 minutes; 95% CI, 40 63 minutes) and those without H. pylori infection (50 minutes; 95% CI, 40-68 minutes; p = 0.85), or between the patients with (median, 45 minutes; 95% CI, 40 68 minutes) and those without (60 minutes; 95% CI, 40-63 minutes; p = 0.77) anxiety. CONCLUSIONS: Orocecal transit time is within the normal range in patients with functional dyspepsia. Anxiety and H. pylori infection do not seem to influence orocecal transit time in these patients. PMID- 12080222 TI - Accuracy of the initial endoscopic diagnosis in the discrimination of gastric ulcers: is endoscopic follow-up study always needed? AB - BACKGROUND: Endoscopic follow-up study of gastric ulcers has been recommended routinely because of the possibility that a gastric neoplasm will be missed in the initial endoscopy. Some authors, most of them reporting data from areas of low gastric carcinoma incidence, have questioned this policy because of the low numbers of curable cancers detected and the high cost of such a program. GOALS: To assess the accuracy of endoscopy diagnosis of gastric ulcers, and to evaluate the efficacy and cost of a gastric ulcer follow-up endoscopic program in an area with an intermediate incidence rate of gastric cancer. STUDY: A retrospective study was used to identify all the gastroscopies in which a gastric ulcer had been diagnosed during a 6-year period. The endoscopic impression was compared with the histologic diagnosis, sensitivity, specificity, positive and negative predictive values, and the likelihood ratio. Patients who completed a follow-up program also were reviewed. For each neoplasm discovered, the number of endoscopies and global cost were calculated. RESULTS: In the 741 gastroscopies performed, 547 gastric ulcers were diagnosed in 529 patients. Biopsies were taken in 330 patients, in whom 341 gastric ulcers were found. At the index endoscopy, 41 gastric neoplasms (12.4%) were diagnosed. The accuracy of endoscopic malignancy diagnosis was as follows: positive predictive value of 0.68, negative predictive value of 0.98, sensitivity of 0.82, and specificity of 0.95. The likelihood ratio was 16. A total of 117 patients completed the follow-up program. Three new cases of gastric cancer (2.6%) were identified. In these three cases, the initial opinion of the endoscopist was uncertain. In the authors' experience, the cost of each gastric cancer diagnosed has been $4.653 (U.S. dollars). CONCLUSIONS: The endoscopic impression correlates with the histologic diagnosis even in a area of intermediate gastric cancer incidence. Endoscopic follow-up study may be restricted to cases of uncertain or malignant endoscopic impression. PMID- 12080223 TI - Should gastric cancer patients more than 80 years of age undergo surgery? Comparison with patients not treated surgically concerning prognosis and quality of life. AB - This study investigated the performance status, mental status, and prognosis of 24 patients older than 80 years whose gastric cancer had been managed surgically during the past 6 years, and 21 patients who were observed conservatively. The advantages and disadvantages of surgery for elderly patients with gastric cancer is discussed. The 3-year survival rate for patients with advanced gastric cancer was 31% for those surgically treated and 0% for those observed conservatively. The difference is statistically significant. The survival rate for patients with early gastric cancer was 62.5% in the surgical treatment group and 33.3% in the conservative observation group, a difference that is not significant. Decline in performance status and deterioration of mental status after open surgery were slight, and the survival rate for patients treated surgically was significantly higher than for conservatively observed patients, regardless of performance status and whether mental status had deteriorated. On univariate and multivariate analyses, the presence or absence of open surgery and macroscopic classification were the only significant prognostic factors, whereas performance status and the presence or absence of mental deterioration were not significant prognostic factors. These results suggest that surgical treatment should not be discouraged even for patients older than 80 years. PMID- 12080224 TI - Expression of e-cadherin and catenins in early gastric cancer. AB - E-cadherin and its associated cytoplasmic proteins including alpha-, beta-, and gamma-catenin play a pivotal role in the maintenance of normal tissue architecture and the suppression of cancer invasion. The purpose of this study was to evaluate the expression of E-cadherin and alpha-, beta-, and gamma-catenin in a larger sample of early gastric cancer, and to examine the relation between these expressions and various clinicopathologic variables. The expression of E cadherin and alpha-, beta-, and gamma-catenin was investigated using immunohistochemical technique with formalin-fixed, paraffin-embedded tissue specimens obtained from 108 patients who underwent surgery for early gastric cancer. In the gastric mucosa of noncancerous areas, epithelial cells showed equally strong membranous expression of E-cadherin and alpha-, beta-, and gamma catenin proteins at the cell-cell boundaries. Reduced expression of E-cadherin and alpha-, beta-, and gamma-catenin was demonstrated in 43.5%, 39.8%, 42.6%, and 50% of cancer tissues, respectively. Whereas 34 tumors (31.5%) displayed preserved expression of all four E-cadherin-catenin complex components, 21 tumors (19.4%) displayed reduced expression of all components of this complex. Reduced expression of E-cadherin and alpha- and gamma-catenin occurred more frequently in diffuse than in intestinal types of cancer, and decreased expression of E cadherin and alpha-, beta-, and gamma-catenin correlated with poor differentiation. The expression of E-cadherin and beta- and gamma-catenin did not correlate with the patient's age, gender, tumor size, location, macroscopic type, depth of invasion, or lymph node metastasis. Only reduced expression of alpha catenin correlated with lymph node metastasis. Reduced expression of all four E cadherin-catenin complex components correlated with poorly differentiated and diffuse-type cancers, but not with the patient's age, gender, tumor size, location, macroscopic type, depth of invasion, or lymph node metastasis. These results suggest that dysfunction of the E-cadherin-catenin complex occurs in an early stage of carcinogenesis, playing a crucial role in disruption of tissue architecture and loss of differentiation in early gastric cancer. PMID- 12080225 TI - A rare case of ulcerative colitis complicating Wilson's disease: possible association between the two diseases. AB - A case of ulcerative colitis complicated by Wilson's disease is described. In this case, ulcerative colitis occurred 12 years after the diagnosis of Wilson's disease, and the colitis was intractable to prednisolone and salazosulfapyridine. Because copper is one of the trace elements necessary for antioxidant defenses during inflammatory process, altered copper metabolism may have contributed to the intractability of the ulcerative colitis in this case. PMID- 12080226 TI - Control of alkaline reflux esophagitis after total gastrectomy by a percutaneous jejunostomy tube. AB - Postoperative alkaline reflux esophagitis is a potentially devastating complication after total gastrectomy. The advent of the Roux-en-Y reconstruction has significantly decreased the incidence of this complication. However, when reflux esophagitis occurs, it is often refractory to medical treatment. Even though surgical revision of the Roux-en-Y anastomosis is the management option of choice, affected patients (especially those with advanced metastatic cancers) are often poor surgical candidates. We describe a novel treatment of refractory alkaline reflux esophagitis in a patient after radical total gastrectomy with Roux-en-Y reconstruction for advanced gastric carcinoma. Radiologic placement of a percutaneous jejunostomy tube into the proximal jejunal limb resulted in significant symptomatic relief by external diversion of the pancreaticobiliary drainage, and the procedure was associated with minimal discomfort. Nonsurgical pancreaticobiliary diversion should be considered in those patients with refractory alkaline reflux esophagitis that occurs after total gastrectomy. PMID- 12080227 TI - Evaluation of factors that have reduced mortality from acute pancreatitis over the past 20 years. AB - BACKGROUND: The mortality associated with acute pancreatitis varies markedly in different studies, with most frequently reported mortality rates of 10% to 15% for all cases and 15% to 90% for attacks regarded as "severe." More recently, various centers have recorded lower mortality rates of 4% to 7% for all attacks of acute pancreatitis and 20% to 50% for those regarded as severe. GOALS: To investigate whether there has been a reduction in mortality associated with acute pancreatitis over the past 20 years and the reasons for this reduction. STUDY: Intended as a review, this study included the authors' 20-year prospective assessment of mortality as it relates to the severity of the disease, complications, and current therapy. For the mortality results, the study was divided into four 4-year periods from 1977 to 1998 and the past 3 years (i.e., 1998-2001). For comparison, the mortality figures from some other large studies are presented. RESULTS: This study showed that the initial reduction in mortality related to acute pancreatitis coincided with the recognition and application of the signs of severity, either Ranson's prognostic signs or Bank's clinical criteria. These signs dictated admission to intensive care unit (ICU) therapy, the intensity of ICU monitoring, and the importance of organ-specific emergent therapy. Further mortality reduction in the 1990s could be attributed to either a more select study sample or earlier and more selective endoscopic or surgical debridement of infected tissue, endoscopic cyst drainage, and angiographic control of gastrointestinal bleeding. Improved nutritional support by jejunal feeding, earlier use of antibiotic therapy, gut sterilization, early endoscopic retrograde cholangiopancreatography for common bile duct stones and necrosectomy for noninfected necrosis have reduced the overall mortality associated with acute pancreatitis to a mean of 5% (range, 3.8-7%) for all cases and 20% (range, 15 25%) for severe cases. However, it is clear that the greater the number of signs denoting severity of organ failure, the higher the mortality. CONCLUSIONS: There has been considerable reduction in the mortality associated with acute pancreatitis over the past 20 years. The reasons are multifactorial, but recognition of severity signs, early implementation of organ-specific therapy, and newer endoscopic, surgical, and angiographic therapy for infection cyst and bleeding appear to have been the major factors in reducing mortality. PMID- 12080228 TI - Tropical pancreatitis. AB - Tropical pancreatitis is an uncommon cause of acute, and often chronic, relapsing pancreatitis. Patients present with abdominal pain, weight loss, pancreatic calcifications, and glucose intolerance or diabetes mellitus. Etiologies include a protein-calorie malnourished state, a variety of exogenous food toxins, pancreatic duct anomalies, and a possible genetic predisposition. Chronic cyanide exposure from the diet may contribute to this disease, seen often in India, Asia, and Africa. The pancreatic duct of these patients often is markedly dilated, and may contain stones, with or without strictures. The risk of ductal carcinoma with this disease is accentuated. Treatment may be frustrating, and may include pancreatic enzymes, duct manipulations at endoscopic retrograde cholangiopancreatography, octreotide, celiac axis blocks for pain control, or surgery via drainage and/or resection. PMID- 12080229 TI - Proposal for a new grading system for chronic pancreatitis: the ABC system. AB - BACKGROUND: A satisfactory classification, or staging system, for chronic pancreatitis is not available. Currently available systems use histologic proof or imaging data for assessment. Although the former is not available in all cases, the latter may not correlate well with the clinical presentation. Therapy for chronic pancreatitis depends on the clinical aspects of the disease, not purely on the duct morphology or histology. GOALS: A staging/grading system is proposed that is applicable to all types of diagnosed chronic pancreatitis and that allows comparison of similar groups. It also provides therapeutic guidelines. The system divides patients into grades A, B, and C, depending on the presence or absence of abdominal pain or complications. Further subdivision is based on the presence or absence of deficiency in pancreatic function. This staging system permits accurate description of the disease with relevance to therapeutic approach and prognosis. PMID- 12080230 TI - Prevalence and pathogenesis of duodenal ulcer in chronic alcoholic pancreatitis. AB - BACKGROUND: The prevalence of duodenal ulcer (DU) has been considered high in patients with chronic pancreatitis; however, its pathogenesis is unclear. We hypothesized that Helicobacter pylori infection plays the major pathogenetic role. STUDY: One hundred seven cases (97 men, 10 women) of chronic alcoholic pancreatitis (CAP) were prospectively investigated from 1997 to 2001. One hundred thirty-seven DU patients and 59 nonulcer dyspepsia patients formed the two control groups. Pancreatic function was evaluated by determination of fecal fat excretion and fasting blood glucose concentration. Upper gastrointestinal endoscopy was performed in all patients, and gastric mucosal biopsies were taken for assessment of H. pylori infection with a modified Giemsa stain and rapid urease test. RESULTS: Fifteen (14%) of the 107 patients with CAP had active DU. There was a trend toward an association between the presence of diabetes mellitus and/or steatorrhea and the occurrence of DU in patients with CAP (p = 0.06). The rate of H. pylori infection was significantly higher in patients with CAP and DU than in those with only CAP (86.7% vs. 54.3%, p = 0.02) but the rate similar to that in patients with simple DU (75.2%). Trends toward higher prevalence of H. pylori infection in CAP with DU were noticed when they were compared with the nonulcer dyspepsia group (86.7% vs. 66.1%). There was no significant difference in prevalence of H. pylori between CAP patients without DU and dyspeptic patients (54.3% vs. 66.1%). CONCLUSIONS: These data demonstrate that the prevalence of DU in CAP is relatively high. H. pylori infection seems to play the major pathogenetic role in DU associated with CAP. PMID- 12080231 TI - Prognosis of symptomatic versus asymptomatic autoimmune hepatitis: a study of 68 patients. AB - BACKGROUND: Autoimmune hepatitis (AIH) is a chronic liver disorder of unknown etiology. Disease presentation ranges from asymptomatic to symptomatic onset, fulminant, acute, or chronic. GOALS: To evaluate the prognosis of patients with asymptomatic versus symptomatic AIH and to determine the role of early treatment and prognostic factors in this subgroup of patients. METHODS: Sixty-eight patients with AIH were retrospectively evaluated for clinical presentation, liver function tests, autoantibody profile, liver biopsy, treatment, and long-term prognosis. RESULTS: Twenty-three patients were classified as having asymptomatic AIH. They were compared with the 45 AIH patients with symptomatic onset. Patients with symptomatic and asymptomatic presentations were indistinguishable by age, but there was a relative male predominance in the asymptomatic group. Aminotransferase and immunoglobulin levels were lower in the asymptomatic group. In contrast, the histopathologic picture was indistinguishable between the two groups. Lobular hepatitis of a moderate to severe degree, portal fibrosis, and bridging fibrosis were detected in similar frequencies in both groups. Asymptomatic onset led to a delay in diagnosis and, consequently, in the initiation of treatment and was associated with use of relatively lower doses of corticosteroids. Response to treatment and long-term prognosis of asymptomatic patients was better than that observed in the symptomatic group. CONCLUSIONS: Asymptomatic onset of AIH is relatively common. The clinical presentation appears in correlation with liver functions tests but not with liver histology. This subgroup of AIH carries a better response to treatment and a favorable prognosis. PMID- 12080233 TI - A survey of current liver biopsy practice patterns. AB - GOALS: To survey clinicians regarding current liver biopsy practice patterns. BACKGROUND: Although the hepatitis C epidemic has increased the proportion of hepatology in general gastroenterology practice, many clinicians express concern regarding the risks of percutaneous liver biopsy. STUDY: A questionnaire about liver biopsy practices was sent to members of the Duke University Digestive Epidemiological Studies Consortium. RESULTS: The response rate was 112 (71%) of 157. Thirty-three (29.5%) physicians reported that they do not perform liver biopsies. Reasons cited for not performing biopsies included concern about risks (72.7%), low reimbursement (66.7%), and logistical issues with space and recovery time (45.4%). Routine practice was biopsy without ultrasound in 53.2%, ultrasound marking by a radiologist or technician at the time of biopsy in 24.0%, previous ultrasound marking in 17.7%, and ultrasound marking by the gastroenterologist in 5.1%. For patients with hepatitis C, 76.8% of clinicians perform routine biopsies before treatment. CONCLUSIONS: A significant proportion of clinicians do not perform liver biopsies; common reasons include the risks of the procedure and the low reimbursement. The use of ultrasound with liver biopsy has become more common. New approaches, especially in training programs, may be necessary to make clinicians more comfortable with this procedure. PMID- 12080232 TI - Treatment of hepatitis C virus genotype 4-related cirrhosis: ribavirin and interferon combination compared with interferon alone. AB - BACKGROUND: Response to treatment with interferon alfa, with or without concomitant ribavirin, varies with the viral genotype and the degree of fibrosis in patients with chronic hepatitis C virus (HCV). GOALS: To determine the response of HCV type 4-related cirrhosis to interferon and ribavirin combination treatment compared with interferon alone. STUDY: Patients living in Kuwait were assigned to take either interferon alone at a dosage of 5 million units thrice weekly (26 patients) or interferon 5 million units thrice weekly combined with ribavirin 1,000 mg/d (21 patients) for 24 weeks. Biochemical response was defined as normal alanine aminotransferase (ALT) at end of therapy. Sustained biochemical response was defined as normal ALT 6 months after the end of therapy. Sustained virologic response was defined as negative serum HCV RNA 6 months after the end of therapy. RESULTS: Only 2 (8%) of 26 patients showed biochemical response after interferon alone, whereas 11 (52%) of 21 showed biochemical response after interferon combined with ribavirin (p < 0.01). Only 2 (8%) of 26 patients showed sustained biochemical response after interferon alone, whereas 5 (23%) of 21 showed sustained biochemical response after interferon combined with ribavirin (not significant, p > 0.1). None of the 26 patients showed virologic response after interferon alone, whereas 3 (14%) of 21 showed sustained virologic response after interferon combined with ribavirin (not significant, p > 0.1). CONCLUSION: These results suggest that patients with cirrhosis caused by HCV type 4 show no response to interferon alone and only slightly better response to 24 weeks of interferon combined with ribavirin. PMID- 12080234 TI - Nephrotoxicity of interferon alfa-ribavirin therapy for chronic hepatitis C. AB - The nephrotoxicity associated with interferon therapy for chronic hepatitis C infection has not been clearly defined. We describe a patient with chronic hepatitis C infection who developed the nephrotic syndrome during treatment with interferon and ribavirin. Renal biopsy revealed focal segmental glomerulosclerosis. She had a virologic and biochemical response to the antiviral therapy, and the nephrotic syndrome improved after termination of antiviral therapy. We place our case report in context with a review of the literature on nephrotoxicity associated with interferon therapy. Because our patient had no other obvious reason for the nephrotic syndrome, we are postulating that it may be secondary to interferon-ribavirin therapy. The temporal relation between the administration of the drug and the detection of toxic affects (nephrotic syndrome) and subsequent improvement upon withdrawal also supports a causative role for interferon-ribavirin. Although nephrotoxicity is rare, it should be emphasized that it can occur anytime after the start of interferon therapy, and physicians treating patients with chronic hepatitis C must be aware of this idiosyncratic, unpredictable, and potentially serious adverse event. PMID- 12080235 TI - Metastatic peripheral primitive neuroectodermal tumor (PNET) masquerading as liver abscess: a case report of liver metastasis in orbital PNET. AB - Adult primitive neuroectodermal tumor (PNET) of the orbit is an extremely rare malignant tumor. We report the case of a 37-year-old woman with PNET metastasis of the liver 3 years after treatment of the primary right intraconal orbital PNET with resection and chemoradiation adjuvant therapy. Literature review revealed eight previous cases of orbital PNET, but this is the first case report of liver metastasis arising from orbital PNET. PMID- 12080236 TI - Response to article by Buchman et al. Use of hyperbaric oxygenation in the treatment of ulcerative colitis. PMID- 12080237 TI - Estrogen and cyproterone acetate combination-induced autoimmune hepatitis. PMID- 12080238 TI - Isoniazid-induced acute hepatitis and acute pancreatitis in a patient during chemoprophylaxis. PMID- 12080239 TI - Wilson's disease with neuropsychiatric manifestations and liver disease but no Kayser-Fleischer ring. PMID- 12080240 TI - Is pain experienced during liver biopsy an important factor? PMID- 12080241 TI - Fatal adrenal metastasis from hepatocellular carcinoma. PMID- 12080242 TI - Constrictive pericarditis presenting as chylous ascites. PMID- 12080243 TI - Angiotensin-converting enzyme inhibitors and diarrhea. PMID- 12080244 TI - Carcinoid syndrome from gastrointestinal carcinoid tumor without distant metastases. PMID- 12080246 TI - A case of intraductal papillary mucinous tumor of pancreas combined with early gastric cancer and MALToma of stomach. PMID- 12080245 TI - Concomitant triple lesions of adenocarcinoma, malignant lymphoma, and granular cell tumor of the stomach. PMID- 12080252 TI - Laparoscopic suture closure of perforated duodenal peptic ulcer. AB - The aim of this study was to assess the outcome of a continuous series of 30 patients with perforated duodenal peptic ulcers treated by a laparoscopic approach. Between January 1996 and December 1998, 30 patients (24 males, 6 females) with a mean age of 69.2 years were operated on with a laparoscopic approach. Laparoscopic treatment included peritoneal lavage, suture of the perforation, and omental patching in 24 cases. A conversion to laparotomy was necessary in five patients (16.6%). Mean operative time was 92 minutes (range: 58 114) and mean hospital stay was 6 days (range: 4-16). Mortality and morbidity rates were 6.6% (n = 2) and 16.6% (n = 5). With a median follow-up of 12 months, 96% of the patients were in good condition; one patient had recurrent duodenal ulceration. The results of our study show the feasibility of the laparoscopic approach for perforated peptic ulcer repair, with acceptable mortality and morbidity rates. PMID- 12080253 TI - Laparoscopic choledochoduodenostomy: review of a 4-year experience with an uncommon problem. AB - A laparoscopic choledochoduodenostomy (LCDD) may be performed when the common bile duct (CBD) is obstructed by primary or secondary stones or strictures. A biliary bypass procedure has two goals in view. The short-term goal is complete removal of stones and bypass of obstruction and stricture to restore biliary drainage. The long-term goal is preventing a recurrence of the problem. There is debate over the superiority of any one procedure to achieve both goals. Therefore, it may help the practicing clinician to be aware of the success (or failure), on a case-by-case basis, of these procedures. This awareness may help in the choice of technique. To date, since 1991, we have performed 16 LCDDs; however, in this report, we describe our results with LCDD over the last 4 years to emphasize the usefulness of this procedure. We find that it is a safe and effective procedure for treating patients with benign bile duct obstruction, even for those whose condition may be described as complicated or difficult. Evidence is slowly accumulating that LCDD is also successful in promoting long-term biliary drainage. We reviewed our LCDDs done over the past 4 years, documenting our preoperative, intraoperative, and postoperative experience. A successful LCDD was performed on all six patients. None of the patients had postoperative leaks. There was only one death, which was due to the patient's comorbidities and not the procedure itself. The hepatobiliary enzyme levels returned to normal in all of the surviving patients. The average postoperative length of stay was 6 days. With proper selection and adequate laparoscopic experience, LCDD can be performed in a safe and effective way. PMID- 12080254 TI - Value and consequences of routine intraoperative cholangiography during cholecystectomy. AB - Since the introduction of laparoscopic cholecystectomy (LC), an increase in accidental common bile duct (CBD) injuries of up to 1.2-1.6% has been reported. In the present prospective study of 1,710 patients undergoing cholecystectomy (1,241 LC procedures and 469 open cholecystectomies [OC]), we tested the predicative value of routine intraoperative cholangiography (IOC). The IOC was feasible in 92.4% of the cases in the LC group and in 83% of cases in the OC group and presented a complete depiction of the extrahepatic bile system in 98.3%. Anatomic variations of the bile duct system, which influenced the operative management, were found in 13.2% of cases (13.4% LC versus 12.8% OC). In 2.5% of the patients, preoperatively undetected CBD stones were also found. Method-specific complications did not occur in any of the patients. Additionally, in a controlled subgroup analysis of 163 patients, we evaluated preoperative intravenous cholangiography (IVC) and IOC. Intravenous cholangiography showed only 72.4% of the operation-relevant anatomic variations (vs. 100% by IOC); in 6.1% of the cases, there were reactions to the dye (vs. none in IOC), and in only 28.6% of the patients, CBD stones were detected (vs. 71.4% IOC). There were four bile duct injuries (0.29%) during LC and two (0.4%) during OC. All injuries were detected intraoperatively and fixed in the same setting without postoperative complications. In conclusion, we recommend the use of routine IOC during cholecystectomy. By this technique, anatomic variations of the bile duct system will be visualized and therefore accidental injuries will be avoided. PMID- 12080255 TI - Hand-assisted laparoscopic ultrasound-guided radiofrequency thermal ablation of liver tumors: a technical report. AB - Ultrasound-guided radiofrequency thermal ablation has been performed for liver tumors by percutaneous, laparoscopic, or open surgical approaches. Each approach has specific advantages and disadvantages. Herein we describe a new technique for hand-assisted laparoscopic ultrasound-guided radiofrequency thermal ablation of liver tumors. A hand-access device is placed at the right or central portion of the abdomen, in addition to standard trocars. A conventional intraoperative ultrasound probe, with an ultrasound guidance system attached, is inserted into the peritoneal cavity together with the surgeon's hand. After pneumoperitoneum is established, an electrode-cannula for thermal ablation is introduced subcostally or intercostally, and advanced into a liver tumor under direct guidance by intraoperative ultrasound. We have used this technique in eight patients with unresectable liver tumors. Precise guidance of the cannula into tumors was possible. All tumors were well ablated. The postoperative recovery of patients was of shorter duration compared with that of an open surgical approach. A hand assisted laparoscopic ultrasound-guided method has advantages of both laparoscopic and open surgical approaches for radiofrequency thermal ablation treatment of liver tumors. Accurate cannula insertion is possible with the ultrasound guidance system. The hand-assisted laparoscopic approach can become an additional useful technique, particularly as a valuable alternative to an open surgical method, for performing radiofrequency thermal ablation. PMID- 12080256 TI - Enteroclysis-guided laparoscopic adhesiolysis in recurrent adhesive small bowel obstructions. AB - The aim of this study was to point out the efficiency of enteroclysis assay in localization of intraabdominal adhesions that impede small bowel transit in patients with recurrent adhesive small bowel obstruction who underwent laparoscopic partial adhesiolysis. Between January 1998 and June 2001, 15 selected patients with recurrent adhesive small bowel obstructions were treated successfully by medical means and evaluated with enteroclysis to define the pathologic adhesive site that impeded bowel transit. If the results of enteroclysis were indicative, they underwent laparoscopic partial adhesiolysis. The mean duration of the laparoscopic procedure was 99 minutes. In one patient conversion to laparotomy occurred because of excessive adhesions, and in another patient a small bowel injury occurred and enterorrhaphy was performed laparoscopically. Mean postoperative hospital stay was 4 days. During a mean follow-up of 17.2 months (range, 6-39), there was no delayed morbidity or recurrence. Identification of the small bowel site of recurrent obstruction with enteroclysis permits limited laparoscopic adhesiolysis. This approach may be a rational alternative to not only open procedures but also complete laparoscopic adhesiolysis without enteroclysis. PMID- 12080257 TI - Outpatient laparoscopic incisional/ventral hernioplasty: our experience in 55 cases. AB - Laparoscopic repair of ventral and incisional hernias is still a controversial therapeutic option. The purpose of this article is to evaluate the results of laparoscopic surgery on ventral hernias in an outpatient surgery unit of a university hospital. Fifty-five patients consecutively underwent laparoscopic surgery for ventral/incisional hernias in the outpatient unit. The patients' clinical features, hernia type, intraoperative, and postoperative complications and reasons for hospital admission are studied. Forty-two patients (76%) were discharged on the day of the surgery. Thirteen (24%) required hospital admission; 9 admissions were predictable (5 for intraoperative occurrences and 4 for associated surgery) and 4 were unpredictable (all for uncontrolled pain; 31%). We found no case of hospital admission for vomiting, urinary retention, or dizziness. Statistical analysis of the patients requiring admission showed no significant correlation with their clinical features or with the site, size, or recurrence of the abdominal wall defect (P > 0.05). Laparoscopic repair of ventral and incisional hernias can be done as a highly efficient ambulatory procedure, and morbidity and hospital admission with this technique do not depend on the type of hernia. PMID- 12080258 TI - Big-screen laparoscopic surgery. AB - Significant advances in fiberoptic and digital technology for laparoscopic surgery have been made over the past decade. One area that appears to be overlooked in this field is the advancement in the display of the image during laparoscopic surgery. The authors describe the use of digital video-cinema equipment as a simple and effective technique that enhances the projection of the surgical view. This method has been found to be visually more comfortable, aiding the surgical procedure, and extremely useful as a teaching tool. PMID- 12080259 TI - Color coding of sutures in laparoscopic perforated duodenal ulcer: a new concept. AB - A simple modification for securing the omental patch during laparoscopic oversewing of perforated duodenal ulcers is presented. This new concept uses color coding of sutures to prevent confusion and suture "cut-out" during the tie over of the omental patch. The concept of suture color-coding may be applicable to other laparoscopic procedures where identification of "suture pairs" is difficult. PMID- 12080260 TI - Laparoscopic reduction of an acute gastric volvulus and repair of a hernia of Bochdalek. AB - We report a case of an acute strangulated gastric volvulus in a hernia of Bochdalek in an adult female patient that was repaired successfully via the laparoscopic approach. A left-sided diaphragmatic hernia contained a strangulated but viable gastric volvulus and a noncompromised colon. The contents of the hernia were reduced, and the 4-cm congenital diaphragmatic defect was primarily repaired with nonabsorbable sutures. The patient was discharged on the second postoperative day and remained symptom-free at 7 months. Unlike the very few previous reports of elective laparoscopic repair of uncomplicated Bochdalek hernias, this appears to be the first report of an urgent laparoscopic repair of a complicated hernia of this type. PMID- 12080261 TI - Calcified bile duct stone not crushable by endotriptor or mechanical lithotriptor: case report. AB - When we remove bile duct stones in endoscopic retrograde cholangiopancreatography, we sometimes encounter the complication of basket impaction. In most cases, bile duct stones can be crushed with a mechanical lithotriptor. An endotriptor also is commonly used to resolve the problem of basket impaction. An endotriptor is more powerful than a mechanical lithotriptor in crushing stones. We report a case of basket impaction that was not resolved by means of an endotriptor. When abdominal radiography shows apparent calcified bile duct stone, it should be cautioned that the stone is sometimes too hard to be crushed, even with use of the endotriptor as well as a mechanical lithotriptor. PMID- 12080262 TI - New technique for finding the ruptured bile duct into the liver cysts: scope in the cave technique. AB - Communication with the biliary tree is the most frequent complication of hepatic hydatid disease. This may result in fistula formation after surgical management of liver hydatidosis. Although frank ruptures are usually diagnosed preoperatively and managed surgically, simple openings may result in biliary fistula formation. It is very difficult to accurately diagnose a small hole in the cyst during surgery in most patients. We describe an easy and reliable technique for finding the point of communication via direct visualization using a telescope during conservative surgery for hepatic hydatidosis. PMID- 12080263 TI - Laparoscopy-assisted resection of complicated Meckel's diverticulum in adults. AB - The era of videoendoscopic surgery in emergency surgery practice has facilitated a wide range of endoscopic operative procedures. In our unit the diagnosis of acute abdomen is made after sequential clinical and laboratory examination, and diagnostic laparoscopy is advocated. Laparoscopy-assisted resection of complicated Meckel's diverticulum in two adult patients was performed, and the results are discussed. Between December 1996 and June 2000, 98 patients underwent diagnostic laparoscopy at the Cerrahpasa Medical Faculty Emergency Surgery Unit of Istanbul University because of signs and symptoms of acute abdomen. Among these, we have diagnosed one case of intestinal obstruction due to a volvulus around Meckel's diverticulum and one of acute abdomen masquerading as acute appendicitis but actually involving omphalomesenteric duct cyst torsion. Both of these patients underwent diagnostic laparoscopy and laparoscopy-assisted Meckel's diverticulectomy. Diagnostic laparoscopies were performed on 46 male (47%) and 52 female (53%) patients. In all cases, laparoscopy successfully confirmed the diagnosis. Although in 27 patients the interventions were converted to open procedures, the operations were completed laparoscopically in 71 patients. Two of these patients underwent laparoscopy-assisted Meckel's diverticulectomy and their postoperative periods were uneventful. Both patients were discharged from the hospital on their fourth postoperative day. Diagnostic laparoscopy is a safe and effective method for diagnosis of acute abdomen. In emergency surgery practices in developing countries, advanced laparoscopy should be performed to reduce expenses. Laparoscopy-assisted Meckel's diverticulectomy is a safe and economic procedure and can be performed in adults for treatment of complicated cases without staplers. PMID- 12080264 TI - Diagnostic and therapeutic laparoscopy for trauma: a technique of safe and systematic exploration. AB - Laparoscopy has a limited role in the evaluation of a stable trauma patient. The main concern addressed in the literature is a significant potential for missed visceral injury and a minimal role for therapeutic application. We present a simple technique for systematic abdominal exploration for a suspected intraabdominal injury in a series of three consecutive trauma patients (two penetrating injuries, one blunt) at a level-one trauma center. Three 5-mm ports and a 30-degree laparoscope were used for exploration. Additional ports for retraction were used as needed for therapeutic procedures. All patients underwent complete exploration of the abdominal cavity. Therapeutic procedures consisted of suturing an intraperitoneal bladder rupture, draining a lacerated tail of the pancreas, and cauterizing an actively bleeding penetrating wound to the left lobe and caudate lobe of the liver. All patients experienced prompt and uncomplicated recovery. There were no missed injuries. The presented technique enables a systematic laparoscopic exploration of the abdomen, which follows the same principles as open exploration. The role of laparoscopy in the treatment of a stable trauma patient is likely to increase as more surgeons acquire advanced laparoscopic skills, and as systematic exploration techniques are used. PMID- 12080265 TI - Tribute to Karl Zucker: an ambitious, dedicated, and unselfish surgeon of his time. PMID- 12080272 TI - Laparoscopy-assisted Billroth-I gastrectomy (LADG) for cancer: our 10 years' experience. AB - To evaluate laparoscopy-assisted Billroth-I gastrectomy (LADG), we examined the outcome of its use over the last 10 years. From December 1991 to December 2001, 116 patients with early gastric cancer underwent LADG in the surgical department of Oita Medical University and Koga hospital by the same surgical staffs. An operation record and clinical sheets were reviewed to obtain the operative findings, clinical course, and pathologic findings of resected specimens to evaluate the usefulness of LADG in the management of early gastric cancer. In all LADG procedures, regional lymph nodes dissection (D1+alpha) was successfully performed using laparoscopy. The mean operative duration and blood loss were 234 minutes and 139 mL, respectively. There were only four major complications, including pneumonia, leakage of anastomosis, pancreatic injury, and anastomotic stenosis, but all these cases were successfully treated conservatively. The mean length of postoperative stay was 16.3 +/- 2.5 days. All patients except one, who died not of cancer but of cerebral bleeding, were alive without recurrence or port-site metastasis during mean follow-up period of 45 months. We successfully performed 116 LADG procedures over 10 years. This procedure is recommended for the treatment of patients with early gastric cancer because of the associated good prognosis and several benefits, including less invasiveness and early recovery. PMID- 12080273 TI - Laparoscopic reoperation as management of severe gastroesophageal reflux following laparoscopic Heller myotomy for achalasia: a case report. AB - Achalasia is a rare disorder of the esophagus. Nonsurgical management includes oral medication, pneumatic dilatation, and injections of botulinum toxin. Surgical intervention was traditionally limited to patients with residual dysphagia after nonsurgical treatment. With the popularization of minimally invasive surgery, myotomy was increasingly performed via a laparoscopic approach. The procedure was found to be safe and efficient and is now used with increasing frequency as a primary therapeutic option. We report the case of a 17-year-old patient with achalasia in whom symptoms of gastroesophageal reflux developed following laparoscopic Heller myotomy without an antireflux procedure. Five years after surgery, the patient underwent reoperation with Toupet fundoplication. Five months after surgery, we found a normal De Meester Score and no pathologic gastroesophageal reflux. The authors conclude that laparoscopic Heller myotomy is the treatment of choice for achalasia and recommend that an antireflux procedure be included routinely. PMID- 12080274 TI - Yasmin--an oral contraceptive with a new progestin. PMID- 12080275 TI - New treatments for actinic keratoses. PMID- 12080278 TI - Effects of morphine on tumour growth. AB - Endogenous opiate alkaloids, such as morphine, and their peptide counterparts have been implicated in a wide variety of pharmacological and physiological functions. In addition to their use in the treatment of pain, opioids, appears to be important in the growth regulation of normal and neoplastic tissue. This review will focus on the influence of endogenous and exogenous opioids on tumour growth, with emphasis on immunoregulatory and antiproliferative mechanisms. PMID- 12080279 TI - The role of stress in neurodegenerative diseases and mental disorders. AB - OBJECTIVE: Evidence for a connection between stress and selected neurodegenerative diseases as well as mental disorders is analyzed. Does stress cause or exacerbate related pathophysiological disease processes? METHOD: The stress phenomenon is illustrated and the impact of stress on the nervous system, neurodegenerative diseases, and mental disorders is examined. The connection between stress and the hippocampus - and its association with memory functions - is described. In particular, the pathophysiological significance of stress in Alzheimer's disease, multiple sclerosis, anxiety, depression, posttraumatic stress disorder, and schizophrenia is investigated. RESULTS: Stress plays a major role in various (patho)physiological processes associated with neurodegenerative diseases and mental disorders. In principle, stress has the potency to exert either ameliorating or detrimental effects. The specific outcome depends on multiple variables. However, the amount of stress experienced in relation to activated physiological processes that aim at successful coping and positive adjustments (i.e., stress response) most often is overwhelming - and may thus become detrimental in the long-term. Moreover, the hippocampus is sensitive to stress, and its involvement in neurodegeneration - in the course of stress related disease processes - may account for severe clinical disabilities (e.g., memory loss). DISCUSSION/CONCLUSION: Stress has a major impact upon neurodegenerative diseases and mental disorders. It plays a significant role in susceptibility, progress, and actual outcome. Also, subjective or individual differences have to be taken into account. However, stress - especially 'adequate' acute stress (stress that is not overwhelming) - may even improve performance/biological functions and be beneficial in certain cases. PMID- 12080280 TI - Cellular localization of a chromogranin B-like derived peptides in leeches. AB - Using immunocytochemistry techniques, we demonstrated specific immunostaining with antibodies rose against fragment (547-560) and (614-626) of bovine chromogranin B (CGB) at the level of brain and the tegument of the leech Theromyzon tessulatum. The used of these antibodies on leech sections revealed immunopositive labeling in neurons and glial cells of the central nervous system (CNS), and epidermal glandular cells of the tegument. Co-localization between two antibodies rose against different fragment of bovine CGB have been demonstrated in neurons and glial cells of leech CNS like in vertebrates. Finally, the whole of the data showed for the first time the presence in leeches of CGB like derived peptides. PMID- 12080281 TI - The effect of pyridoxine administration on melatonin secretion in normal men. AB - OBJECTIVES: To determine pineal response to pyridoxine in normal men. MATERIAL AND METHODS: Twelve healthy men were given orally pyridoxine (100 mg) or placebo at 1700h. Serum melatonin levels were determined every 30 minutes with simultaneous measurement of core body temperature between 1700h to 0300h. Polysomnographic sleep recordings were performed between 1800h to 2000h. RESULTS: Serum melatonin levels after both placebo and pyridoxine showed a nocturnal rise occurring at 22:10+/-1:22h and 22:24+/-1:09h, respectively. The melatonin onset, peak, mean and area under the curve (AUC) values after pyridoxine (3.2+/-1.6 pg/ml, 47.2+/-22.6 pg/ml, 31.5+/-11.0 pg/ml and 173.5+/-138.4 pg/ml x min, respectively) were similar to the values after placebo administration (4.7+/-1.6 pg/ml, 53.9+/-26.0 pg/ml, 37.2+/-2.8 pg/ml and 205.3+/-137.8 pg/ml x min, respectively). CBT revealed a significant nocturnal decline but without significant difference between pyridoxine and placebo. Sleep amount and architecture were similar after the two treatments. CONCLUSIONS: In adult man, the oral administration of 100 mg-pyridoxine during the evening hours has no effect on melatonin secretion nor does it alter CBT or sleep quality. PMID- 12080282 TI - Early prediction and psycho-immunologic mediation of minor illness in adulthood. AB - BACKGROUND: The Barker Hypothesis suggests that an unfavourable uterine environment can have the effect of programming the body for disease later in life. Research indicates a bidirectional relationship between thought and biochemical reactions, that may be influenced by early programming. Reports suggest that 25% of variance in birthweight is a result of foetal environment and that the health and cognitive deficits do not just affect those with an officially low birthweight. OBJECTIVE: This study investigates the influence of birthweight on cognition and minor illness in adults. METHODS: This is a retrospective, cross sectional design with an opportunity sample of 75 adults. Participants whose birthweight ranged from 2.5 kg to 4.88 kg, completed a symptom check list and general self-efficacy scale, reporting on the previous month. RESULTS: Analysis of variance indicates that those with higher birthweight have fewer minor illness symptom days and higher general self-efficacy. Regression analysis indicates that birthweight is significantly predictive of levels of minor illness and general self-efficacy. CONCLUSION: From the findings of this and previous studies, it is possible to infer vertical coactions between foetal environment and immuno competence. It is suggested that birthweight is an early predictor of levels of a cognitive mediator and minor illness. Data were applied to an equilibrium model to represent the relationship in terms of Gottlieb's concept of horizontal and vertical coactions. PMID- 12080283 TI - Event related P300 potential in NIDDM patients without cognitive impairment and its relationship with previous hypoglycemic episodes. AB - OBJECTIVES: The purpose of this study was to evaluate the ERP P300 in non insulin dependent diabetes mellitus (NIDDM) patients without cognitive impairment and the relationship with clinical variables, the presence of retinopathy and previous hypoglycemic episodes. METHODS: NIDDM patients (N=44) without evidence of cognitive impairment and controls (N=17) were studied clinically and with ancillary exams and the ERPs P300 were recorded. Patients were examined clinically and with the Folstein Mini-Mental Examination (MMSE) for cognitive function and all patients showed a score higher than 26 (maximal value=30). Previous hypoglycemia was evaluated through a questionnaire establishing the number of episodes and the symptoms of hypoglycemia in a scale scoring from zero to 15. RESULTS: ERP P300 latencies were significantly higher in NIDDM patients than in controls (p<0.03). ERP P300 measures were significantly related to age (Pearson, p<0.01) and not to metabolic variables, disease duration or the presence of retinopathy. Severity of hypoglycemia was not associated to ERP P300 latency. CONCLUSIONS: Our study supports the evidence that NIDDM patients, without signs of nervous system involvement, have ERP P300 alterations and this is not related to retinopathy, metabolic variables or previous hypoglycemic episodes. Chronic hyperglycemia may alter brain glucose transport and increase tolerance to hypoglycemia effects in the nervous system. PMID- 12080284 TI - Hypoxia alters testis development in neonatal rats. AB - OBJECTIVE: To investigate the effects of continued hypoxia on postnatal development of the rat testis. DESIGN: Wistar rats were exposed to simulated hypoxia from birth to postnatal day (PND) 45. Testosterone (T) in the plasma and the testis was measured in rats at PND 3, 7, 14, 21, 28, 35 and 45 respectively. Testis histology and cellular ultrastructure were studied. RESULTS: a) Hypoxia induced a significant arrest of testes weight gain after PND 28; b) T release was significantly attenuated after PND 21; c) Alterations in histology and cellular ultrastructure were found in the testis, showing the swelling of testis interstitium and the enlargement of mitochondria in Leydig cells. CONCLUSION: Postnatal hypoxia stress alters testis development both in terms of function and structure, especially at the critical age of gonadal development. PMID- 12080285 TI - Circadian rhythm of melatonin in patients with colorectal carcinoma. AB - OBJECTIVE: The aim of the study was pineal gland function assessment on the base of daily rhythm study and mean daily melatonin (MEL) concentrations in serum in patients with colorectal carcinoma. MATERIAL AND METHODS: Studies were performed in 12 women at the age of 63.17+/-5.90 years and 21 men aged 58.95+/-11.32 years with large intestine adenocarcinoma. The control group consisted of 28 healthy volunteers at comparable age. During the circadian study blood samples for the measurement of melatonin (MEL) were collected every 4 hours during 12 h. MEL concentrations were assessed with the use of RIA methods. Statistical analysis of circadian rhythms of MEL was carried out with the use of cosinor method according to Halberg. RESULTS: Existence of daily rhythm of MEL secretion was shown in all studied groups. A significant decrease of amplitude of rhythm and secretion of MEL at nocturnal hours in comparison with the control group was shown in the group of women with large intestine carcinoma. A significant decrease of mesor value and amplitude of MEL rhythm as a consequence of decrease of MEL secretion at nocturnal and morning hours was observed in the group of ill men. CONCLUSIONS: Decrease in melatonin circadian rhythm amplitude as a consequence of its lowered nocturnal secretion occurred in all patients with colorectal carcinoma. Abnormalities in daily rhythm of melatonin secretion were more intensified in men with large intestine carcinoma, which leads to suppression of mean daily hormone concentration. PMID- 12080286 TI - Circadian rhythm of melatonin in postmenopausal asthmatic women with hormone replacement therapy. AB - OBJECTIVE: The aim of the study was to check if in postmenopausal women such a disease as asthma and use of hormone replacement therapy (HRT) influence daily melatonin (MEL) secretion. MATERIAL AND METHODS: Studies were performed in 55 asthmatics (treated and not treated with glucocorticosteroids) and 20 healthy postmenopausal women (aged 48-60) before HRT and after 6 months of transdermal 17beta-estradiol and medroxyprogesterone acetate treatment (cyclical method). During the circadian study blood samples for the measurement of MEL were collected every 3 hours during the day. MEL concentrations were assessed with the use of RIA methods. Statistical analysis of the circadian rhythm was performed with the use of cosinor test according to Halberg et al. RESULTS: Existence of daily rhythm of MEL secretion was shown in all studied groups, before as well as after HRT. A significant decrease of mesor and amplitude's rhythm was observed in the group of women with asthma treated with glucocorticosteroids (GC) before and after HRT use in relation to women not treated with GC and the control group. Secretion of MEL in these patients was lower at nocturnal hours. A significant decrease of mean daily MEL secretion in relation to values before HRT use was shown in all groups. CONCLUSIONS: Asthmatic postmenopausal women treated with GC show lowered circadian secretion of melatonin as a consequence of lowering its secretion at nocturnal hours. Hormonal replacement therapy causes a decrease of daily melatonin secretion in healthy as well as asthmatic women, not disturbing circadian rhythm of this hormone's secretion. PMID- 12080287 TI - Comparative neuroprotective effects of preischemic PACAP and VIP administration in permanent occlusion of the middle cerebral artery in rats. AB - OBJECTIVES: Pituitary adenylate cyclase activating polypeptide (PACAP) and vasoactive intestinal polypeptide (VIP) belong to the same peptide family, and both neuropeptides have been shown to exert in vitro and in vivo neurotrophic and neuroprotective effects. The aim of the present study was to investigate and compare the protective effects of PACAP and VIP in permanent focal cerebral ischemia in rats. The effect on the progression of the cerebral infarct was also studied. METHOD: Male rats were injected 450 pmol PACAP or VIP dissolved in physiological saline intracerebroventricularly, preceding the occlusion of the middle cerebral artery. Control animals received vehicle treatment. Permanent focal ischemia was induced by the intraluminal filament occlusion of the middle cerebral artery. Animals were sacrificed 12 or 24 hours after the onset of ischemia, and infarcted brain areas were determined by staining bran sections with triphenyl-tetrazolium chloride. RESULTS: Twelve hours after ischemia, the infarcted brain volume resulted to be 14.8% in the control group, 15.3% in the VIP-treated group and 5.8% in the PACAP-treated animals. Twenty-four hours after middle cerebral artery occlusion, the infarcted brain volumes were 21.5%, 20.7% and 14.3% in the control, VIP and PACAP-treated animals, respectively. CONCLUSION: Our results provide further evidence for the neuroprotective effects of PACAP38 as given in form of a preischemic bolus. It slows down the progression of the evolution of the infarct and reduces the final infarct size. In contrast, a related peptide, VIP, does not have neuroprotective effects under the same experimental conditions. PMID- 12080288 TI - A new neuroimmunotherapeutic strategy of subcutaneous low-dose interleukin-2 plus the long-acting opioid antagonist naltrexone in metastatic cancer patients progressing on interleukin-2 alone. AB - OBJECTIVES: Recent advances in knowledge of Psychoneuroimmunology have shown that several neuroactive substances, including neurohormones and neuropeptides, may exert immunomodulatory effects. However, despite the great variety of potential neuroimmune interactions, at present we may recognize two major neuroendocrine systems exerting a physiological neuroimmunomodulatory function, consisting of the pineal gland and the brain opioid system, provided by immunostimulatory and immunosuppressive effects, respectively. Recent in human studies have demonstrated the possibility to amplify the biological activity of IL-2, the major anticancer cytokine, by pineal indoles. MATERIALS & METHODS: The present study was carried out to draw some preliminary in human results on the possible immunomodulatory effects of the inhibition of the brain opioid activity by a long acting opioid antagonist, naltrexone (NTX). The study was performed in 10 metastatic renal cell cancer patients, who had progressed on a previous immunotherapeutic cycle with IL-2 alone. Patients were treated with the same doses of IL-2 (6 million lU/day subcutaneously for 6 days/week for 4 weeks) plus an oral administration of NTX at a dose of 100 mg every 2 days. RESULTS: The clinical response consisted of a partial response in 1 and a stable disease in 5 patients, whereas the other 4 patients progressed. Therefore, the percent of non progressive disease was 6/10 (60%). Moreover, mean lymphocyte increase achieved during IL-2 plus NTX was significantly higher (P<0.05) than that obtained during the previous treatment with IL-2 alone. CONCLUSIONS: This study shows that a blockade of the brain opioid system, which plays a physiological immunosuppressive role, may improve the anticancer effects of IL-2 in humans. PMID- 12080289 TI - Plasma concentrations of 5-HT, 5-HIAA, norepinephrine, epinephrine and dopamine in ecstasy users. AB - Recreational use of the synthetic methamphetamine derivative MDMA (3,4 methylenedioxymethamphetamine), the main constituent of the illegal drug "ecstasy", has increased dramatically in recent years. The reasons for ecstasy associated cardiovascular complications like tachycardia, arrhythmias and hypertensive crises and psychiatric symptoms like psychotic episodes are not well understood. We have measured the plasma concentrations of 5-HIAA, 5-HT, norepinephrine, epinephrine and dopamine in 159 ecstasy users and controls. Ecstasy users showed elevated resting sympathetic activity, reflected in increased norepinephrine, epinephrine and dopamine levels. The levels of these catecholamines correlated positively with the cumulative dose and also with consumption during the last 30 days and 12 months. Although it is known that significant changes in 5-HT and 5-HIAA appear in the cerebrospinal fluid in ecstasy users, we could not detect alterations in serotonergic neurotransmitters in plasma in this large sample of subjects. Thus, in the drug-free interval, ecstasy users show lowered central serotonergic activity (lowered 5-HT and 5-HIAA concentrations in CSF) along with unchanged central noradrenergic and dopaminergic activity (HVA and MHPG unchanged in CSF) and elevated peripheral noradrenergic, dopaminergic and adrenergic activity along with unchanged peripheral serotonergic activity (plasma levels). We conclude, that the data presented here could argue for a noradrenergic hyperreactivity in the drug-free interval in ecstasy users resulting from previous ecstasy consumption. Also for an association with psychotic episodes and cardiovascular complications like tachycardia, arrhythmias. PMID- 12080291 TI - Real doctors, treating real patients. PMID- 12080292 TI - Orthodontists in good company. PMID- 12080293 TI - Orthodontics justified. PMID- 12080294 TI - Orthodontic faculty. PMID- 12080295 TI - Orthodontists rely on education, experience, and research. PMID- 12080296 TI - International Symposium on Early Orthodontic Treatment. Phoenix, Arizona, February 8-10, 2002. PMID- 12080297 TI - Biological rationale for early treatment of dentofacial deformities. PMID- 12080298 TI - Treatment timing and outcome. PMID- 12080299 TI - Preadolescent Class II problems: treat now or wait? PMID- 12080300 TI - Early treatment of skeletal open bite malocclusions. PMID- 12080301 TI - Stability of open bite treatment. PMID- 12080302 TI - Treatment of crowding in the mixed dentition. PMID- 12080303 TI - Early intervention in the transverse dimension: is it worth the effort? PMID- 12080304 TI - Serial extraction revisited: 30 years in retrospect. PMID- 12080305 TI - Stability and relapse: early treatment of arch length deficiency. PMID- 12080306 TI - Biomechanics of maxillary expansion and protraction in Class III patients. PMID- 12080307 TI - Early application of chincap therapy to skeletal Class III malocclusion. PMID- 12080308 TI - Early treatment for impacted maxillary incisors. PMID- 12080310 TI - Autogenic dental transplants. PMID- 12080309 TI - Early treatment of tooth-eruption disturbances. PMID- 12080311 TI - Congenitally missing teeth: orthodontic management in the adolescent patient. PMID- 12080312 TI - Skeletal Class II patterns in the primary dentition. AB - The aim of this longitudinal cephalometric study was to investigate early characteristics of favorable and unfavorable growth. Two groups of untreated subjects with skeletal Class II pattern were formed on the basis of an ANB angle at least 2.5 degrees above the individual norm, according to Jarvinen, at age 5. The subjects were followed longitudinally until age 12. The favorable growth group (n = 13) consisted of subjects who exhibited a decrease in the individualized ANB angle of at least 1.5 degrees during follow-up. The unfavorable growth group (n = 10) was characterized by an increase in the individualized ANB angle during the growth period from 5 to 12 years. Early characteristics of unfavorable growth included increased mandibular plane angle (measured to the sella-nasion line and the palatal plane, P <.05), increased lower gonial angle (P <.05), and a decreased ratio of posterior-to-anterior face height (P <.05) [corrected]. At the longitudinal follow-up at age 12, the 2 groups showed significant differences in SNB (P <.01) and individualized ANB (P <.01). The improved sagittal jaw relationship in the favorable growth group was the result of changed mandibular position. Morphogenetic posterior rotation seems to occur early in patients with skeletal Class II pattern and unfavorable growth and might be part of an adaptive mechanism compensating for insufficient mandibular growth. PMID- 12080314 TI - Rigid versus wire fixation for mandibular advancement: skeletal and dental changes after 5 years. AB - The bilateral sagittal split osteotomy (BSSO) is the most common surgical procedure for the correction of mandibular retrognathism. Commonly, the proximal and distal segments are fixated together with either wire or rigid screws or plates. The purpose of this study was to compare long-term (5 years) skeletal and dental changes between wire and rigid fixation after BSSO. In this multisite, prospective, randomized clinical trial, the rigid fixation group received three 2 mm bicortical position screws, and the wire fixation group received inferior border wires and 6 weeks of skeletal maxillomandibular fixation with 24-gauge wires. Cephalometric films were obtained 2 weeks before surgery and at 1 week, 8 weeks, 6 months, 1 year, 2 years, and 5 years after surgery. Linear cephalometric changes were referenced to a cranial base coordinate system. Before surgery, both groups were comparable with respect to linear and angular measurements of craniofacial morphology. Both groups underwent similar surgical changes. Skeletal and dental movements occurred in both groups throughout the study period. Five years after surgery, the wire group had 2.2 mm (42%) of sagittal skeletal relapse, while the rigid group remained unchanged from immediately postsurgery. Surprisingly, at 5 years, both groups had similar changes in overbite and overjet. This was attributed to dental changes in the maxillary and mandibular incisors. Although rigid fixation is more stable than wire fixation for maintaining the skeletal advancement after a BSSO, the incisor changes made the resultant occlusions of the 2 groups indistinguishable. PMID- 12080313 TI - Masticatory performance and areas of occlusal contact and near contact in subjects with normal occlusion and malocclusion. AB - This preliminary study evaluated relationships between masticatory performance and areas of interocclusal distance contact (<50 microm) and near contact (50-350 microm) of the buccal segments during maximum intercuspation. The sample included subjects with normal occlusion (n = 18) and Class I (n = 14), Class II (n = 13), and Class III (n = 6) malocclusions. Chewing performance was evaluated on the basis of the breakdown of CutterSil (Heraeus Kulze, South Bend, Ind); chewing ability was assessed by the number of chews necessary to swallow jerky and almonds. Impressions of the buccal segments, taken with Blu Mousse (Parkell Bio Materials, Farmingdale, NY) impression material, were scanned and enlarged, and each subject's first molars and premolars were manually traced bilaterally to estimate the platform area. The areas of contact and near contact (ACNC) that measured between 0 and 350 microm thick were estimated optically on the basis of the amount of light transmitted through the impression. The results showed no significant differences in platform area between the right and left sides or between the malocclusion groups. ACNC were negatively related to median particle size and broadness of particle distribution. There were no correlations between ACNC and the number of chews necessary to swallow jerky or almonds. Subjects with normal occlusion had significantly larger ACNC than those with Class I, Class II, and Class III malocclusions, in descending order. Subjects with Class III malocclusions had the smallest areas of near contact (<350 microm). We concluded that ACNC are similar on the right and left sides; that subjects with larger ACNC are better able to break down foods; and that subjects with malocclusions have smaller ACNC than those with normal occlusions. PMID- 12080315 TI - The effect of functional occlusal forces on orthodontic tooth movement and tissue recovery in rats. AB - The influence of physiologic occlusal forces on tissue response to orthodontic tooth movement and its subsequent recovery was investigated in rats. The mandibular incisors were subjected to tipping and intrusive loads for 2 weeks. In 27 animals, the teeth remained in occlusion, while the incisors were shortened out of occlusion in 29. In each group, some rats were killed at the end of the tooth movement process, and the rest were kept alive for a 13-week recovery period. The subgroups were matched with corresponding controls. Tooth intrusion and eruption were monitored throughout the experiment; then histomorphometric and histopathologic evaluations were performed on the incisors and their periodontal tissues. Tooth movement was barely affected by the functional forces. However, immediately upon 2-week load application, damage to the pulp was more extensive in the occluding teeth; this might be due to the combined intrusive vectors of functional and mechanical loads. The effect of the physiologic function was expressed primarily during the recovery period, in which the eruptive function and distorted periodontal ligament space returned to normal significantly faster in the occluding teeth. This was also true for the healing of dental and periodontal lesions. The incidence of tooth resorption was also reduced. Normal occlusal function is imperative for rapid reconstitution of the damage caused by orthodontic forces. PMID- 12080316 TI - Midsymphyseal distraction osteogenesis for correcting transverse mandibular discrepancies. PMID- 12080317 TI - Periodontal concerns associated with the orthodontic treatment of impacted teeth. AB - Impacted teeth are common and are often treated with orthodontic eruption, but periodontal problems associated with the process can evade detection. Profound destruction of the periodontium of an impacted tooth or adjacent teeth can occur. This case report describes the orthodontic eruption of 4 impacted canines in a 19 year-old woman. An open surgical approach was used. Within 6 months of treatment, the maxillary right canine and the lateral incisor experienced severe periodontal destruction, resulting in questionable prognoses for the teeth. Plaque control, periodontal architecture, and subgingival microflora were examined as local etiologic factors of periodontal destruction associated with orthodontic eruption of impacted teeth. Plaque control measures were evaluated, and the consequences of orthodontic tooth movement in the presence of inadequate plaque control were considered. Areas of periodontal architecture made vulnerable by the surgical exposure of the impacted teeth were identified, and the effect of orthodontic force on the periodontium was explored. An increase in putative periopathogens in the subgingival microflora after orthodontic appliance placement was observed. Microbiologic monitoring for pathologic levels of periopathogens and antibiotic therapy were considered. Orthodontic treatment of impacted teeth might require additional professional and personal plaque control measures, 3-dimensional diagnostic imaging, and control of putative periopathogens to preserve the health of the periodontium. PMID- 12080318 TI - Four applications of reinforced polyethylene fiber material in orthodontic practice. AB - Four different orthodontic applications of a polyethylene fiber material are shown and discussed in case reports. Ribbond (Ribbond Inc, Seattle, Wash), a bondable, reinforced polyethylene fiber material, was successfully used in a fixed orthodontic retainer, a space maintainer, a temporary postorthodontic fixation device to attach a pontic to abutment teeth during periodontal therapy, and a posttraumatic stabilization splint. This clinical practice was based on earlier studies showing that the polyethylene fiber material adapted easily to dental contours and could be manipulated during the bonding process. It also has acceptable strength because of good integration of fibers with composite resin; this leads to good clinical longevity. Because thinner composite resins can be used, the volume of the splint can be minimized. There is no need for significant tooth structure removal, so the technique is reversible and conservative. It also satisfies patients' esthetic expectations. These cases show that Ribbond is a promising splint material for use in orthodontic practice. PMID- 12080319 TI - Online patient communications. PMID- 12080320 TI - Litigation, legislation, and ethics: Minors and statutes of repose. PMID- 12080321 TI - Peroxisome proliferator-activated receptor-gamma and esophageal cancer. PMID- 12080322 TI - A nomogram to predict C282Y hemochromatosis. AB - Genetic testing of hemochromatosis has not been widely used as a diagnostic test because of unawareness of its existence and concerns about genetic discrimination. We developed a nomogram for the prediction of C282Y homozygotes for hemochromatosis from transferrin saturation and ferritin using Bayes theorem. The results of transferrin saturation and C282Y genotyping were available for 8,572 participants (5,042 men, and 3,530 women). The study group included patients in population-screening projects, referred cases, and family members. Likelihood ratios were calculated for transferrin saturation in predicting C282Y homozygotes. Pretest probabilities were estimated on the basis of serum ferritin concentration, and a predictive nomogram for men and women was created with the use of Bayes' theorem. In the highest-risk region of the nomogram in men, the probability of C282Y hemochromatosis was 89.7% (95% confidence interval = 85.1 94.3); in the lowest-risk zone it was 1.1% (0.4-1.9). The corresponding regions in women were 88.9% in the high zone (95% confidence interval = 77.0-100.0) and 6.5% in the lowest (95% confidence interval = 4.9-8.1). This approach allows the clinician to predict the probability of a patient's being a C282Y homozygote over a wide range of ferritin and transferrin saturation values instead of above a particular threshold. PMID- 12080323 TI - Granulocyte colony-stimulating factor primes NADPH oxidase in neutrophils through translocation of cytochrome b(558) by gelatinase-granule release. AB - Granulocyte colony-stimulating factor (GCSF) primes reduced neutrophil nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity in response to formyl peptide but does not increase oxidase activity when used alone. Both oxidase activity and degranulation require phospholipase D (PLD) activation, and exogenous C(2)-ceramide inhibits both functions through inhibition of PLD activity. We extended these observations to investigate neutrophil responses to GCSF. GCSF at a dosage of 30 to 100 ng/mL, a concentration range that primes superoxide release, stimulated a 60% to 100% increase in gelatinase release from tertiary granules but did not stimulate lactoferrin release from secondary granules. A 75% to 100% dose-dependent increase in PLD activity in GCSF-treated neutrophils was also observed. Gelatinase release and PLD activity were inhibited by 10 micromol/L C(2)-ceramide. The increase in gelatinase release in response to priming concentrations of GCSF suggests that tertiary granules contribute a component of the NADPH oxidase to the plasma membrane. Neutrophils treated with 50 ng/mL GCSF were found to contain 20% more cytochrome b(558) in the plasma membrane fraction than unstimulated cells, consistent with degranulation of only tertiary granules. Correspondingly, in the presence of 10 micromol/L C(2) ceramide, cytochrome b(558) content in the plasma membrane did not increase after neutrophil activation. In contrast, GCSF did not lead to p47phox translocation to the plasma membrane or phosphorylation. Because phosphorylation and translocation of p47phox are required for oxidase activity, these findings account for the inability of GCSF alone to generate the respiratory burst. We conclude that translocation of cytochrome b(558) was responsible for GCSF priming of NADPH oxidase in neutrophils. PMID- 12080324 TI - Growth inhibition of esophageal squamous carcinoma cells by peroxisome proliferator-activated receptor-gamma ligands. AB - The growth of human cancer cells expressing peroxisome proliferator-activated receptor-gamma (PPAR-gamma) has been reported to be inhibited by PPAR-gamma ligands. In esophageal squamous-cell carcinoma cell lines T.T, T.Tn, and EC-GI 10, we detected expression of PPAR-gamma and investigated the effects of PPAR gamma ligands on these cell lines in vitro with the use of troglitazone, pioglitazone, and 15d-PGJ2. Marked growth inhibition by the PPAR-gamma ligands was observed in all cases. The growth-inhibitory effect was evidenced by a dose dependent inhibition of deoxyribonucleic acid synthesis and was associated with altered cell-cycle progression manifesting G1 arrest. Cell-cycle arrest in T.Tn cells induced by troglitazone could be correlated with an increased level of cyclin-dependent kinase inhibitor p27(Kip1), p21(Cip1/Waf1), and p18(Ink4c). Moreover, troglitazone treatment increased the expression of interleukin-1 alpha, a multifunctional cytokine implicated in antitumor immunity. These findings suggest that troglitazone and other PPAR-gamma ligands have adjuvant or neoadjuvant therapeutic potentials in esophageal cancer. PMID- 12080325 TI - A selective alpha(2)-adrenergic agonist for cardiac resuscitation. AB - The effects of selective alpha(2)-adrenergic agonist alpha-methylnorepinephrine on the initial success of resuscitation and postresuscitation myocardial function were compared with nonselective alpha- and beta-adrenergic epinephrine in a swine model of cardiac arrest. Epinephrine, the primary pharmacological intervention in the treatment of cardiac arrest, improves immediate outcome. However, epinephrine increases the severity of myocardial dysfunction after cardiac resuscitation. Both inotropic and chronotropic actions provoke disproportionate increases in myocardial oxygen consumption by the ischemic heart, prompting this study, in which we hypothesized that a selective alpha(2)-adrenergic agonist, alpha methylnorepinephrine (alpha-MNE), would moderate these adverse effects of epinephrine and minimize postresuscitation myocardial dysfunction. After 7 minutes of untreated ventricular fibrillation (VF) in 14 anesthetized male domestic pigs, precordial compression at a fixed rate of 80 compressions/min was begun, along with mechanical ventilation. Either alpha-MNE (100 microg/kg) or epinephrine (20 microg/kg) was administered as a bolus after 2 minutes of precordial compression. After an additional 4 minutes of precordial compression, defibrillation was attempted. Left ventricular systolic and diastolic function was quantitated with the use of transesophageal echo-Doppler imaging. Comparable increases in coronary perfusion pressure to 15 mm Hg were observed after the administration of both drugs. All animals were successfully resuscitated; epinephrine and alpha-MNE were equally quick in restoring spontaneous circulation after 7 minutes of untreated VF. Ejection fraction was reduced by 35% and 14% by epinephrine and alpha-MNE, respectively, after resuscitation. Epinephrine and alpha-MNE increased postresuscitation heart rate by 38% and 15%, respectively. Accordingly, significantly less postresuscitation impairment followed the administration of alpha-MNE. alpha-MNE, a selective alpha-adrenergic agonist, was as effective as epinephrine in restoring spontaneous circulation after 7 minutes of untreated VF in a porcine model for CPR and demonstrated lesser postresuscitation myocardial injury. PMID- 12080326 TI - Cholesterol metabolism in normal and heterozygous familial hypercholesterolemic newborns. AB - In heterozygous familial hypercholesterolemia (FH), serum low-density lipoprotein (LDL) cholesterol levels are frequently increased in utero. A unique Finnish FH population, FH-North Karelia (FH-NK), has been identified, providing an excellent opportunity to study the diagnostic significance of cholesterol metabolism in FH. For that purpose, we investigated lipoprotein lipids, cholesterol precursors (squalene, methyl, and demethyl sterols), cholestanol, and plant sterols in FH-NK newborns (n = 5), non-FH siblings (n = 7), and controls (n = 20) at birth and after 1-year follow-up in 8 FH-NK and 5 non-FH children. The sum of concentrations (micrograms per deciliter) of methyl sterol (8-monomethylsterol, methostenol, 8-dimethylsterol, 8,24-dimethylsterol, and lanosterol) and squalene was higher in FH newborns than in non-FH siblings but overlapped with one control case. Cord-blood total or LDL cholesterol values could not be used for diagnostic purposes, whereas 1-year LDL cholesterol values were highly superior to those measured at birth. The methyl sterol ratio in cord blood was 29 to 193 10(2) mmol/mol cholesterol and was undetectable in serum at the age of 1 year; those of the demethyl precursor sterols were 1.5 to 8 times higher in cord blood than in serum at the age of 1 year, suggesting that cholesterol synthesis was markedly increased at birth. Plant sterols, not synthesized in human beings, were already present in serum of all the groups at birth, indicating their transfer, apparently with cholesterol, from mother to fetus. Babies born to FH mothers showed a greater tendency toward accelerated cholesterol synthesis than did those born to FH fathers. Despite signs of markedly high but similar synthesis of cholesterol at birth in FH and non-FH newborns, the diagnosis of FH was questionable by measurement of cholesterol precursors or LDL cholesterol in cord blood. The latter measurement, at the 1-year mark, is superior for diagnostic purposes. PMID- 12080327 TI - Expression of monocyte chemoattractant protein-1 in proximal tubular epithelial cells in a rat model of progressive kidney failure. AB - Impairment of kidney function in various types of glomerular disease is associated with tubulointerstitial changes. Monocyte chemoattractant protein-1 (MCP-1) is up-regulated in the tubulointerstitium and in the glomeruli in many human and experimental kidney disorders. We investigated the localization of MCP 1 expression in a rat model of progressive kidney failure. Male Wistar rats were subjected to subtotal nephrectomy (n = 30) or sham surgery (n = 30). Immunohistochemistry with immunoelectron microscopy and in situ hybridization were used to examine the expression of MCP-1 protein and messenger ribonucleic acid (mRNA) in the kidney, respectively. MCP-1 protein and mRNA were hardly detected in both glomeruli and tubulointerstitium of control rats. However, in the rats subjected to nephrectomy, MCP-1 expression was increased in the tubular cells of the remnant kidney, accompanied by significant macrophage infiltration. MCP-1 was observed mainly in the proximal tubular cells and only weakly in distal tubular cells. No significant expression of MCP-1 protein or mRNA was noted in the glomeruli. Immunoelectron microscopy showed the presence of MCP-1 in the rough endoplasmic reticulum of proximal tubular cells, confirming that MCP-1 is produced in proximal tubular cells. MCP-1 was also observed in endocytic vesicles adjacent to the brush border of proximal tubular cells, suggesting incorporation of MCP-1 from the tubular lumen. Our findings indicate localized expression of MCP-1 in proximal tubular cells in the remnant kidney and suggest that MCP-1 in proximal tubular cells is involved in tubulointerstitial damage in chronic kidney failure associated with glomerular hypertension. PMID- 12080328 TI - Differential gene expression in renal-cell cancer. AB - Renal-cell carcinoma (RCC) is an important cause of morbidity and mortality, and its incidence has been increasing. Malignant transformation is thought to be associated with changes in the expression of several genes, and this alteration in gene expression is believed to be critical to the development of the malignant phenotype. In this study, the expression of about 60,000 genes/expressed sequence tags in clear-cell RCC, normal kidney, and a set of diseased nonmalignant kidneys was determined with the use of the Affymetrix microarray technique, and differences in gene expression were analyzed. Many genes were found to be differentially expressed in these two sample sets. The genes that were expressed greater than four times more in RCC, those expressed only in RCC, and those expressed greater than two times more in RCC and also expressed in a limited number of other tissues were analyzed for their expression in a variety of other normal and diseased tissues. Some of the genes identified were overexpressed only in RCC among the tissues examined, and some were overexpressed in several other malignant tissues in addition to RCC. Other genes were overexpressed in RCC compared with normal kidney but were also overexpressed in diseased nonmalignant kidney or a variety of other normal tissues. All of the RCC samples could be clustered together, separate from the normal and diseased kidney samples, with the use of the Eisen clustering technique and a set of 50 genes. The observed changes in gene expression in RCC should help further the understanding of the biology of RCC and may be useful in diagnosis, treatment, and imaging. PMID- 12080329 TI - Assessment of risk for sudden cardiac death. PMID- 12080331 TI - Computer-aided design of a PDZ domain to recognize new target sequences. AB - PDZ domains are small globular domains that recognize the last 4-7 amino acids at the C-terminus of target proteins. The specificity of the PDZ-ligand recognition is due to side chain-side chain interactions, as well as the positioning of an alpha-helix involved in ligand binding. We have used computer-aided protein design to produce mutant versions of a Class I PDZ domain that bind to novel Class I and Class II target sequences both in vitro and in vivo, thus providing an alternative to primary antibodies in western blotting, affinity chromatography and pull-down experiments. Our results suggest that by combining different backbone templates with computer-aided protein design, PDZ domains could be engineered to specifically recognize a large number of proteins. PMID- 12080333 TI - What is the point of a database? PMID- 12080332 TI - Structure and catalytic mechanism of the E. coli chemotaxis phosphatase CheZ. AB - The protein CheZ, which has the last unknown structure in the Escherichia coli chemotaxis pathway, stimulates the dephosphorylation of the response regulator CheY by an unknown mechanism. Here we report the co-crystal structure of CheZ with CheY, Mg(2+) and the phosphoryl analog, BeF(3)(-). The predominant structural feature of the CheZ dimer is a long four-helix bundle composed of two helices from each monomer. The side chain of Gln 147 of CheZ inserts into the CheY active site and is essential to the dephosphorylation activity of CheZ. Gln 147 may orient a water molecule for nucleophilic attack, similar to the role of the conserved Gln residue in the RAS family of GTPases. Similarities between the CheY[bond] CheZ and Spo0F [bond]Spo0B structures suggest a general mode of interaction for modulation of response regulator phosphorylation chemistry. PMID- 12080334 TI - Cutting the leash. PMID- 12080335 TI - Intrinsic metal binding by a spliceosomal RNA. PMID- 12080336 TI - Illuminating proteins with Aladan's lamp. PMID- 12080337 TI - Open season on RNA polymerase. PMID- 12080338 TI - Protein complexes take flight. PMID- 12080339 TI - Picture story. A pathogenic family business. PMID- 12080340 TI - A new family of plant transcription factors displays a novel ssDNA-binding surface. AB - The crystal structure of p24, the single-stranded DNA (ssDNA) binding subunit of the plant defense transcription factor PBF-2, has been determined to 2.3 A resolution. p24 is representative of a novel family of ubiquitous plant-specific proteins that we refer to as the Whirly family because of their quaternary structure. PBF-2 is composed of four p24 molecules that interact through a helix loop-helix motif. This interaction produces a central pore, with beta-strands radiating outwards, resulting in a whirligig appearance to the quaternary structure. The noncrystallographic C(4) symmetry arrangement of p24 subunits is novel for ssDNA binding proteins and may explain the binding specificity of PBF 2. This structural arrangement also supports the role of PBF-2 in binding melted promoter regions to modulate gene expression. PMID- 12080342 TI - Endocannabinoids facilitate the induction of LTP in the hippocampus. AB - Exogenous cannabinoids disrupt behavioral learning and impede induction of long term potentiation (LTP) in the hippocampus, yet endogenous cannabinoids (endocannabinoids) transiently suppress inhibitory post-synaptic currents (IPSCs) by activating cannabinoid CB1 receptors on GABAergic interneurons. We found that release of endocannabinoids by a rat CA1 pyramidal cell during this depolarization-induced suppression of inhibition (DSI) enabled a normally ineffective train of excitatory post-synaptic currents (EPSCs) to induce LTP in that cell, but not in neighboring cells. By showing that endocannabinoids facilitate LTP induction and help target LTP to single cells, these data shed new light on the physiological roles of endocannabinoids and may lead to a greater understanding of their effects on behavior and potential clinical use. PMID- 12080341 TI - Critical periods for experience-dependent synaptic scaling in visual cortex. AB - The mechanisms underlying experience-dependent plasticity and refinement of central circuits are not yet fully understood. A non-Hebbian form of synaptic plasticity, which scales synaptic strengths up or down to stabilize firing rates, has recently been discovered in cultured neuronal networks. Here we demonstrate the existence of a similar mechanism in the intact rodent visual cortex. The frequency of miniature excitatory postsynaptic currents (mEPSCs) in principal neurons increased steeply between post-natal days 12 and 23. There was a concomitant decrease in mEPSC amplitude, which was prevented by rearing rats in complete darkness from 12 days of age. In addition, as little as two days of monocular deprivation scaled up mEPSC amplitude in a layer- and age-dependent manner. These data indicate that mEPSC amplitudes can be globally scaled up or down as a function of development and sensory experience, and suggest that synaptic scaling may be involved in the activity-dependent refinement of cortical connectivity. PMID- 12080343 TI - Antagonists of GLU(K5)-containing kainate receptors prevent pilocarpine-induced limbic seizures. AB - Developments in the molecular biology and pharmacology of GLU(K5), a subtype of the kainate class of ionotropic glutamate receptors, have enabled insights into the roles of this subunit in synaptic transmission and plasticity. However, little is known about the possible functions of GLU(K5)-containing kainate receptors in pathological conditions. We report here that, in hippocampal slices, selective antagonists of GLU(K5)-containing kainate receptors prevented development of epileptiform activity--evoked by the muscarinic agonist, pilocarpine--and inhibited the activity when it was pre-established. In conscious rats, these GLU(K5) antagonists prevented and interrupted limbic seizures induced by intra-hippocampal pilocarpine perfusion, and attenuated accompanying rises in extracellular L-glutamate and GABA. This anticonvulsant activity occurred without overt side effects. GLU(K5) antagonism also prevented epileptiform activity induced by electrical stimulation, both in vitro and in vivo. Therefore, we propose that subtype-selective GLU(K5) kainate receptor antagonists offer a potential new therapy for epilepsy. PMID- 12080344 TI - Role of the chemokine SDF-1 as the meningeal attractant for embryonic cerebellar neurons. AB - Migration of neuronal precursor cells from the external germinal layer (EGL) to the internal granular layer (IGL) is a crucial process in the development of the mammalian cerebellar cortex. These cells make up the only precursor population known to migrate away from the surface of the brain. We studied the role of the chemokine stromal-derived factor 1 (SDF-1) in the cerebellar tissue of rats and knockout mice and found (i) that it functions as an attractive guidance cue for neuronal migration and (ii) that its secretion from non-neuronal meningeal tissue is important for controlling the migration of embryonic EGL cells. PMID- 12080345 TI - Lipid products of PI(3)Ks maintain persistent cell polarity and directed motility in neutrophils. AB - In gradients of external chemo-attractant, mammalian neutrophilic leukocytes (neutrophils) and Dictyostelium discoideum amoebae adopt a polarized morphology and selectively accumulate lipid products of phosphatidylinositol-3-OH kinases (PI(3)Ks), including PtdIns(3,4,5)P(3), at their up-gradient edges; the internal PtdIns(3,4,5)P(3) gradient substantially exceeds that of the external attractant. An accompanying report presents evidence for a positive feedback loop that amplifies the gradient of internal signal: PtdIns(3,4,5)P(3) at the leading edge stimulates its own accumulation by inducing activation of one or more Rho GTPases (Rac, Cdc42, and/or Rho), which in turn increase PtdIns(3,4,5)P(3) accumulation. Here we show that interruption of this feedback by treatment with PI(3)K inhibitors reduces the size and stability of pseudopods and causes cells to migrate in jerky trajectories that deviate more from the up-gradient direction than do those of controls. Moreover, amplification of the internal PtdIns(3,4,5)P(3) gradient is markedly impaired by latrunculin or jasplakinolide, toxins that inhibit polymerization or depolymerization of actin, respectively. Thus reciprocal interplay between PtdIns(3,4,5)P(3) and polymerized actin initiates and maintains the asymmetry of intracellular signals responsible for cell polarity and directed motility. PMID- 12080347 TI - Distinct roles for cyclins E and A during DNA replication complex assembly and activation. AB - Initiation of DNA replication is regulated by cyclin-dependent protein kinase 2 (Cdk2) in association with two different regulatory subunits, cyclin A and cyclin E (reviewed in ref. 1). But why two different cyclins are required and why their order of activation is tightly regulated are unknown. Using a cell-free system for initiation of DNA replication that is based on G1 nuclei, G1 cytosol and recombinant proteins, we find that cyclins E and A have specialized roles during the transition from G0 to S phase. Cyclin E stimulates replication complex assembly by cooperating with Cdc6, to make G1 nuclei competent to replicate in vitro. Cyclin A has two separable functions: it activates DNA synthesis by replication complexes that are already assembled, and it inhibits the assembly of new complexes. Thus, cyclin E opens a 'window of opportunity' for replication complex assembly that is closed by cyclin A. The dual functions of cyclin A ensure that the assembly phase (G1) ends before DNA synthesis (S) begins, thereby preventing re-initiation until the next cell cycle. PMID- 12080346 TI - A PtdInsP(3)- and Rho GTPase-mediated positive feedback loop regulates neutrophil polarity. AB - When presented with a gradient of chemoattractant, many eukaryotic cells respond with polarized accumulation of the phospholipid PtdIns(3,4,5)P(3). This lipid asymmetry is one of the earliest readouts of polarity in neutrophils, Dictyostelium discoideum and fibroblasts. However, the mechanisms that regulate PtdInsP(3) polarization are not well understood. Using a cationic lipid shuttling system, we have delivered exogenous PtdInsP(3) to neutrophils. Exogenous PtdInsP(3) elicits accumulation of endogenous PtdInsP(3) in a positive feedback loop that requires endogenous phosphatidylinositol-3-OH kinases (PI(3)Ks) and Rho family GTPases. This feedback loop is important for establishing PtdInsP(3) polarity in response to both chemoattractant and to exogenous PtdInsP(3); it may function through a self-organizing pattern formation system. Emergent properties of positive and negative regulatory links between PtdInsP(3) and Rho family GTPases may constitute a broadly conserved module for the establishment of cell polarity during eukaryotic chemotaxis. PMID- 12080348 TI - Nucleophosmin regulates the stability and transcriptional activity of p53. AB - Nucleophosmin (NPM) is a ubiquitously expressed nucleolar phosphoprotein that continuously shuttles between the nucleus and cytoplasm. It has been proposed to function in ribosomal protein assembly and transport, and also as a molecular chaperone that prevents proteins from aggregating in the crowded environment of the nucleolus. The NPM gene is involved in several tumour-associated chromosome translocations, which have resulted in the formation of fusion proteins that retain the amino terminus of NPM, including NPM ALK, NPM RAR and NPM MLF1 (ref. 6). It is generally thought that the NPM component is not involved in the transforming potential of these fusion proteins, but instead provides a dimerization interface for the oligomerization and the oncogenic conversion of the various NPM partners (ALK, RAR, MLF1). Here we show that NPM interacts directly with the tumour suppressor p53, regulates the increase in stability and transcriptional activation of p53 after different types of stress, and induces p53-dependent premature senescence on overexpression in diploid fibroblasts. These findings indicate that NPM is a crucial regulator of p53 and suggest that alterations of the NPM function by NPM fusion proteins might lead to deregulation of p53 in tumours. PMID- 12080349 TI - The role of NFAT transcription factors in integrin-mediated carcinoma invasion. AB - Integrins, receptors for extracellular matrix ligands, are critical regulators of the invasive phenotype. Specifically, the alpha(6)beta(4) integrin has been linked with epithelial cell motility, cellular survival and carcinoma invasion, hallmarks of metastatic tumours. Previous studies have also shown that antagonists of the NFAT (nuclear factor of activated T-cells) family of transcription factors exhibit strong anti-tumour-promoting activity. This suggests that NFAT may function in tumour metastasis. Here, we investigate the involvement of NFAT in promoting carcinoma invasion downstream of the alpha(6)beta(4) integrin. We provide evidence that both NFAT1, and the recently identified NFAT5 isoform, are expressed in invasive human ductal breast carcinomas and participate in promoting carcinoma invasion using cell lines derived from human breast and colon carcinomas. NFAT1 and NFAT5 activity correlates with the expression of the alpha(6)beta(4) integrin. In addition, the transcriptional activity of NFAT5 is induced by alpha(6)beta(4) clustering in the presence of chemo-attractants, resulting in enhanced cell migration. These observations show that NFATs are targets of alpha(6)beta(4) integrin signalling and are involved in promoting carcinoma invasion, highlighting a novel function for this family of transcription factors in human cancer. PMID- 12080350 TI - Mismanaging the gift of life: noncompliance in the context of adult stem cell transplantation. AB - In this paper, we address the issue of noncompliance with medical regimens in adult stem cell transplant patients. We review the definition, prevalence, and consequences of noncompliance and address whether or not we should alter our medical decision-making with regard to stem cell transplantation based on patient noncompliance. We discuss how the health care team should handle noncompliance issues and propose clinical guidelines for stem cell transplant programs to consider regarding evaluation and management of medical compliance. PMID- 12080351 TI - Modulations of dose intensity of doxorubicin and cyclophosphamide in association with G-CSF and peripheral blood stem cells in adjuvant chemotherapy for breast cancer: comparative evaluation of completion and safety of three intensive regimens. AB - The aim of this study was to evaluate and to compare in terms of toxicity the modulations of dose intensity of cyclophosphamide and doxorubicin in adjuvant chemotherapy for high-risk breast cancer. Four cycles of sequential high-dose chemotherapy with doxorubicin and cyclophosphamide (AC), supported with G-CSF and peripheral blood stem cells (PBSC) were administered to 81 women. Three successive cohorts were studied: doxorubicin (75 mg/m(2)) + cyclophosphamide (3000 mg/m(2)) every 21 days (group 1), doxorubicin (75 mg/m(2)) + cyclophosphamide (3000 mg/m(2)) every 15 days (group 2), and doxorubicin (75 mg/m(2)) + cyclophosphamide (6000 mg/m(2)) every 21 days (group 3). Seventy-five patients received four cycles of treatment with a total of 310 cycles administered. The received dose intensity of doxorubicin was higher in group 2 and that of cyclophosphamide was lower in group 1 than in the other two groups. Hematological and extra-hematological toxicities, as well as the number and duration of hospitalizations for toxicity, were significantly higher in group 3. We conclude that the group 3 regimen is associated with toxicities comparable to autologous transplantation. Increasing dose intensity of doxorubicin and cyclophosphamide is feasible in an outpatient setting and safe in groups 1 and 2 with the support of hematopoietic factor and PBSC. PMID- 12080352 TI - Use of anti-BDCA-2 antibody for detection of dendritic cells type-2 (DC2) in allogeneic hematopoietic stem cell transplantation. AB - TH2-inducing dendritic cells (DC2) are commonly identified as negative for lineage markers and positive for HLA-DR and CD123 expression. More recently, normal blood DC2 were shown also to be positive for BDCA-2 and BDCA-4 antigens. The aim of this study was to evaluate whether BDCA-2 expression on DC2 is impaired in patients undergoing an allogeneic hematopoietic stem cell transplantation (HSCT) and in healthy donors treated with G-CSF for HSC mobilization. Flow cytometry assays for DC2 detection using either a triple staining with anti-HLA-DR PerCP, anti-Lin(+) anti-CD34 FITC and anti-CD123 PE monoclonal antibodies (mAbs), or a double staining with anti-HLA-DR PE and anti BDCA-2 FITC mAbs were compared in blood samples from patients who underwent an allogeneic HSCT (n = 30) or from healthy donors before (n = 11) and after (n = 8) G-CSF mobilization, as well as in healthy donors' leukapheresis products (n = 12) or bone marrow (n = 4). Staining of BDCA-2(+) cells with other markers such as anti-CD38, anti-CD54 and anti-CD58 were also performed. Median values of CD123(+) DC2 and BDCA-2(+) DC2 were not statistically different in the blood of patients previously treated with chemotherapy, nor in the blood or bone marrow of heathy donors. Also, a 5 day G-CSF treatment did not affect BDCA-2 or adhesion molecule expression on healthy donors' blood DC2 significantly. A correlation between all the results (n = 65) obtained with the two assays was demonstrated in a linear regression curve (r = 0.914) (P = 0.00001). BDCA-2 is a marker highly specific for DC2 that is not downregulated by chemotherapy or G-CSF treatment. Therefore, the anti-BDCA-2 mAb can be efficiently combined with other mAbs and used in studies addressing the role of DC2 in the allogeneic HSCT setting. PMID- 12080353 TI - A randomized trial of assessment of efficacy of leukapheresis volumes, 8 liters vs 12 liters. AB - It is logical to expect that large-volume leukapheresis may be able to collect adequate numbers of PBSC with fewer procedures. To date, there is no agreement on the optimal volume of leukapheresis. Therefore, in this study we compared 8 l volume with 12 l and assessed whether a 50% increase in the blood volume processed would decrease the number of leukaphereses each patient needed to collect > or =2.5 x 10(6) CD34(+) cells/kg in normal mobilizers. PBSC mobilization was done with cyclophosphamide etoposide followed by rhG-CSF in all patients. Forty patients were randomized to undergo 8 l leukaphereses (n = 20 patients) or 12 l leukaphereses (n = 20). The median numbers of leukaphereses required in order to collect > or =2.5 x 10(6) CD34(+) cells/kg in patients processed with 8 l and 12 l were 1 (range 1-5) and 1 (1-4), respectively (P = 0.50). The median number of total nucleated cells (TNC) collected per patient was greater for the 12 l group (7.47 x 10(8)/kg vs 3.90 x 10(8)/kg, P < 0.001), as was the median number of total mononuclear cells (TMNC) (4.26 x 10(8)/kg vs 2.16 x 10(8)/kg, P < 0.001), whereas there was no difference between the two groups for the median number of CD34(+)cells collected per patient (8.94 x 10(6)/kg vs 8.60 x 10(6)/kg, P = 0.85). The TNCs and TMNCs collected per leukapheresis were again greater for the 12 l group (3.64 x 10(8)/kg vs 1.91 x 10(8)/kg, P = 0.001 and 2.17 x 10(8)/kg vs 0.88 x 10(8)/kg, P < 0.001), whereas there was no difference between the two groups for the median number of CD34(+) cells collected per leukapheresis (3.98 x 10(6)/kg vs 3.26 x 10(6)/kg, P = 0.90). This study showed that there is no difference between 8 l and 12 l volumes in regard to collected CD34(+) cells/kg and also the use of a 12 l leukapheresis volume did not decrease the number of leukaphereses performed compared with a 8 l leukapheresis volume. In fact, the use of the larger leukapheresis volume had the disadvantage of adding 60 min to the time the patient was on the machine. PMID- 12080354 TI - Contrast media triggering cutaneous graft-versus-host disease. AB - Adverse reactions to iodinated contrast media are varied and known to develop in patients with asthma and a history of allergy. We describe three successful allogeneic bone marrow transplantation (BMT) patients, who all developed dermal graft-versus-host disease (GVHD) after receiving contrast media. Cutaneous GVHD triggered by contrast media has not been reported to date and has implications for the assessment, monitoring and treatment of patients during the post transplant period. PMID- 12080355 TI - Increased risk of complicated CMV infection with the use of mycophenolate mofetil in allogeneic stem cell transplantation. AB - Mycophenolate mofetil (MMF) is increasingly used for prophylaxis and therapy of GVHD in allogeneic stem cell transplantation. In some recent reports of use of MMF in solid organ transplantation a high incidence of CMV disease has been described. We evaluated the frequency and course of active CMV infection in patients who received MMF compared to those who did not receive MMF after allogeneic stem cell transplantation. We retrospectively analyzed 48 adult patients who consecutively underwent unmanipulated allogeneic bone marrow (n = 15) or peripheral stem cell transplantation (n = 33) from HLA-compatible family donors (n = 30) or unrelated donors (n = 18) from February 1997 to September 2000 at our institution. Only patients who were evaluable for the first 100 days were included in this analysis. Sixteen patients received MMF post transplant (MMF+). CMV-antigenemia was monitored by CMV-pp65 antigen. CMV-antigenemia occurred in 14 patients and was virtually only observed in CMV-IgG+ recipients (13/23, 56%). CMV IgG+/MMF+ patients developed a higher incidence of CMV-antigenemia (8/9, 89%) compared to the CMV-IgG+/MMF- patients (5/14, 35%; P < 0.05). Moreover, five of six patients with persistent or recurrent CMV-antigenemia received MMF. No patient in either group developed CMV disease or died of CMV-related complications. In multivariate analysis including MMF treatment, unrelated vs related donor, GVHD, CMV-serostatus of the donor and stem cell graft type, only MMF treatment was found to be a significant risk factor for both overall and complicated CMV infection. PMID- 12080356 TI - Thrombotic thrombocytopenic purpura and hemolytic uremic syndrome following allogeneic bone marrow transplantation. AB - We monitored the levels of various cytokines and chemokines, as well as an adhesion molecule and factors related to vascular endothelial damage, in three patients with thrombotic thrombocytopenic purpura and hemolytic uremic syndrome after bone marrow transplantation. Measurements were done at the onset of this condition and during plasma exchange for treatment. At the onset of thrombotic thrombocytopenic purpura and hemolytic uremic syndrome, the levels of interleukin 8, thrombomodulin, and plasminogen activator inhibitor-1 were all markedly increased. A close relationship was observed between improvement in symptoms by plasma exchange and a decrease in interleukin-8 level, suggesting that this chemokine may be related to the development of thrombotic thrombocytopenic purpura and hemolytic uremic syndrome after bone marrow transplantation. PMID- 12080357 TI - Listeriosis in recipients of allogeneic blood and marrow transplantation: thirteen year review of disease characteristics, treatment outcomes and a new association with human cytomegalovirus infection. AB - Listeriosis is uncommon in recipients of allogeneic blood, marrow and organ transplantation. Six patients with systemic Listeria monocytogenes infection during 1985-1997 at Bone Marrow Transplantation Service, Memorial Sloan-Kettering Cancer Center are described. In two male and four female patients, the median duration from transplantation to isolation of L. monocytogenes was 62.5 (range 29 to 821) days. Among five allogeneic marrow transplant recipients, four (80%) received HLA antigen matched, T cell-depleted grafts from three unrelated and a single related donor. One patient underwent mismatched-related marrow graft transplant. Cord stem cell transplantation was performed in a single patient. Two required therapy for graft-versus-host disease (GVHD). The 13 year incidence of systemic Listeria infections was 0.47 percent. All six presented with fever (>39 degrees C), and L. monocytogenes bloodstream invasion. Mental status changes and meningioencephalitis were observed in two (33.3%). A concurrent primary opportunistic infection was present in five individuals (83.3%), and four (80%) were being treated for acute human cytomegalovirus (HCMV) viremia. Sixty-six percent responded to therapy and two died from unrelated, non-listeric causes. Systemic listeriosis was uncommon in our high-risk allogeneic blood and marrow transplantation population, and response to therapy with parenteral ampicillin and gentamicin was excellent. The association between primary HCMV reactivation and subsequent listeric infection emphasizes the significance of HCMV-related dysfunction in hosts' cellular immune responses, especially in the setting of allogeneic transplantation. PMID- 12080358 TI - Assessment of psychological distress in prospective bone marrow transplant patients. AB - Patient psychological distress is associated with many aspects of the bone marrow transplantation (BMT) process and has been linked with poor treatment outcomes. We assessed psychological distress in potential BMT candidates, and compared patient and nurse coordinator ratings of emotional distress at the time of initial BMT consultation. Fifty patients self-reported psychological distress using both the NCCN Distress Thermometer (DT) and the Hospital Anxiety and Depression Scale (HADS). Coordinators rated patient emotional distress using the DT and Coordinator Rating Scales that measure anxiety and depression. Fifty and 51% of patients self-reported clinically significant levels of emotional distress and anxiety, respectively, but only 20% self-reported clinically significant levels of depression. There was good correlation between ratings using the brief DT and the more comprehensive HADS. There was significant but only moderate agreement between patient and coordinator ratings of emotional distress and anxiety, with coordinators underestimating the number of patients with high levels of emotional distress. In addition, coordinator ratings of patient emotional distress primarily reflected anxiety, whereas anxiety and depression together only minimally accounted for patient self-reports of psychological distress. These findings suggest that: (1) the DT can be a useful screening device; (2) approximately half of patients at the time of initial consultation for BMT already experience significant levels of psychological distress; and (3) coordinators observe emotional distress primarily as anxiety, but patients experience psychological distress as something more than anxiety and depression. PMID- 12080359 TI - Autologous transplantation of CD133 selected hematopoietic progenitor cells in a pediatric patient with relapsed leukemia. AB - A pediatric patient with very early meningeal relapse of his CD34(+) CD133(-) pre B-ALL was transplanted with 2.5 x 10(6)/kg CD133 selected autologous progenitor cells. Enrichment of CD133(+) cells resulted in a purity of 92.3 +/- 3.5% CD133(+). Hematopoietic engraftment with >1.0 x 10(9)/l neutrophils and >50 x 10(9)/l platelets was reached within 13 and 24 days, respectively. At a follow-up of 11(1/2) months after autologous transplantation, the patient is in complete remission. To our knowledge, the successful transplantation with a CD133 selected graft is the first one to be reported worldwide. CD133 selected cells may serve as an alternative in the case of CD34(+) malignancy. PMID- 12080360 TI - Liver failure complicating non-alcoholic steatohepatitis following allogeneic bone marrow transplantation for Shwachman-Diamond syndrome. AB - Bone marrow transplantation is potentially curative therapy for the hematologic complications associated with Shwachman-Diamond syndrome (SDS). This syndrome is, however, also associated with significant pancreatic and hepatic dysfunction, which may complicate BMT. We report a case of liver failure due to non-alcoholic steatohepatitis (NASH) following BMT for SDS. This case illustrates the need for assessing liver dysfunction pre-BMT in these patients, in addition to highlighting the potential risk posed by pre-existing steatosis for the development of rapidly progressive hepatic failure following transplantation. PMID- 12080361 TI - Transplantation for accidental acute high-dose total body neutron- and gamma radiation exposure. AB - Accidental exposure to acute high-dose total body neutron radiation is rare. We report a 35-year-old man exposed to a total body dose of 5.4 Gy neutron- and 8.5 13 Gy gamma-radiation in a radiation criticality accident. He received a blood stem cell transplant from his HLA-identical sister. There was bone marrow recovery with complete donor chimerism. Random chromatid breaks were observed in donor cells suggesting a bystander effect of neutron exposure. The subject died 82 days after the accident (75 days post transplant) from multi-organ failure. PMID- 12080362 TI - Lymphoma patients undergoing high-dose therapy and ABMT, and EBMJ guidelines. PMID- 12080363 TI - Quantitative trait loci affecting a courtship signal in Drosophila melanogaster. AB - Courtship plays a major role in the sexual isolation of species, yet the genetics underlying courtship behaviour are poorly understood. Here we analyse quantitative trait loci (QTL) for a major component of courtship song in recombinant inbred lines derived from two laboratory strains of Drosophila melanogaster. The total variance among lines exceeds that between parental strains, and is broadly similar to that seen among geographic strains of the Cosmopolitan form of this species. Previous studies of the quantitative genetics of fly song have implied a polygenic additive inheritance with numerous genes spread throughout the genome. We find evidence for only three significant QTLs explaining 54% of the genetic variance in total. Thus there is evidence for a few large effect genes contributing to the genetic variance among lines. Interestingly, almost all of the candidate song genes previously described for D. melanogaster do not coincide with our QTLs. PMID- 12080364 TI - Temporal genetic variation in Aedes aegypti populations in Ho Chi Minh City (Vietnam). AB - Aedes aegypti, the main vector of dengue viruses in Asia, displays variation in population density over time. The larval habitats of this species being unevenly distributed and transient (depending on cycles of drought and flood), the forces generating temporal variation in gene frequencies in populations are studied. We sampled seven mosquito populations from Ho Chi Minh City (Vietnam) and its suburbs on five occasions between April 1999 and August 2000. We investigated genetic variation by studying isoenzyme and microsatellite polymorphism and susceptibility to a dengue 2 virus strain. Ae. aegypti populations collected during the dry season (January-April) showed genetic differentiation (F(ST) = 0.016, P < 10(-6) for isoenzymes) and showed more differentiated infection rates of the dengue 2 virus. The genetic structure of the population is less marked during the rainy season (F(ST) = 0.081, P < 10(-6)). Thus, environmental factors, such as rainfall and factors related to human activity, such as breeding site density and insecticide treatment, control the genetic structure of Ae. aegypti populations in the short term. The implications of studies of this kind for the design of future control programmes are discussed. PMID- 12080365 TI - Evolutionary aspects of the ZZ/ZW sex chromosome system in the Characidae fish, genus Triportheus. A monophyletic state and NOR location on the W chromosome. AB - Four species/populations of Triportheus, T. guentheri, T. cf. elongatus and T. paranense from different Brazilian hydrographic basins, were studied cytogenetically. All the species showed a similar karyotypic macrostructure, with a diploid chromosome number 2n = 52 and a ZZ/ZW sex chromosome system. Besides silver- and fluorochrome-staining, the chromosome mapping of 18S rDNA was also investigated using a biotinylated probe. In spite of some variation in the number of the NORs, a major chromosome site was always present on the short arm of an autosomal pair. In addition, a characteristic rDNA site was also observed on the telomeric region of the W chromosome in the four species/populations. In Triportheus differential reduction in size and heterochromatin accumulation appear to be the main processes associated with the evolution of the sex W chromosome. The location of rRNA genes on this chromosome may correspond to a plesiomorphic condition in the genus and, if so, predates to the sex chromosome system differentiation, with a possible influence in the initial steps of this process. PMID- 12080366 TI - Postglacial distribution area expansion of Polyommatus coridon (Lepidoptera: Lycaenidae) from its Ponto-Mediterranean glacial refugium. AB - The genetic population structure of Polyommatus coridon (Poda, 1761) (Chalk-hill blue) was studied by means of allozyme electrophoresis in north-eastern Germany, the Czech Republic, Slovakia and Hungary. All analysed parameters showed high genetic diversity within populations (number of alleles: 2.61; observed and expected heterozygosity: 18.6% and 19.7%, respectively; percentage of polymorphic loci: total: 73.6%, on 95% level: 56.1%), whereas genetic differentiation between populations was comparatively low (F(ST) = 0.028 +/- 0.005 s.d.). Hierarchical variance analysis revealed significant structuring among five regional population clusters. A significant isolation-by-distance structure exists (r = 0.39; P < 0.05). The mean number of alleles per locus declined significantly from south to north and showed a strong correlation with the geographical latitude (r = -0.88, P < 0.0001). We suggest that this reflects the loss of alleles during the postglacial colonisation of eastern Central Europe from an ice-age refugium in the Balkans. A possible scenario for the postglacial expansion process in eastern Central Europe is discussed using these data: coming from the north-western part of the Balkans, P. coridon may have reached the western tip of Hungary, and consecutively colonised eastern Central Europe using two alternative expansion routes. PMID- 12080367 TI - Heterozygote deficiencies in small lacustrine populations of brook charr Salvelinus Fontinalis Mitchill (Pisces, Salmonidae): a test of alternative hypotheses. AB - Empirical studies of natural populations have commonly reported departures from Hardy-Weinberg expected proportions of heterozygote individuals. Recent advances in statistical population genetics now offer the potential to exploit individual multilocus genotypic information to test more rigorously for possible sources of heterozygote deficiencies. In a previous study in lacustrine brook charr (Salvelinus fontinalis), we reported stronger deficits in small than in large lakes. In the present paper, we propose a methodology for empirically testing alternative hypotheses to identify the cause of the deficits observed in three of the smallest lakes (85, 109 and 182 ha) analysed. First, as in several salmonid species, brook charr may exhibit a trophic polymorphism in north temperate lakes. If morphs are genetically divergent, indiscriminate sampling of both forms would result in less heterozygote individuals than expected in a randomly mating population (Wahlund effect). Using an individual-based method aiming at detecting cryptic population structure, we can reject this explanation as the sole source of deficits for all three lakes. Secondly, mating among relatives could also be frequent in small lakes and lead to heterozygote deficiencies. Significantly more fish than expected at random had low individual multilocus heterozygosity in two of the lakes, suggesting that inbred fish may have been present. Thirdly, sampling of genetically related fish would also lead to departures from Hardy Weinberg proportions. In the same two lakes, the distribution of pairwise relatedness coefficients departed from its random expectation, suggesting that non-random sampling of kin may have occurred. PMID- 12080368 TI - Phylogenetic relationships among Spanish Barbus species (Pisces, Cyprinidae) shown by RAPD markers. AB - Random amplified polymorphic DNA (RAPD) markers were used to estimate the population structure and phylogenetic relationships among the eight species of the genus Barbus that inhabit the Iberian Peninsula. Ten random oligodecamers were used to amplify DNA from 232 fish sampled from 15 populations. A total of 270 markers were detected that revealed low levels of genetic variability. The conclusions of cluster analysis indicate two main branches and three well differentiated groups: north-eastern, Mediterranean and Atlantic. This clustering mainly reflects the evolutionary history of the genus, which is closely related to the paleogeography of the Iberian Peninsula. The contribution and application of these results to the conservation of the species, to their taxonomic status and to the process of colonization of the Iberian Peninsula by the genus Barbus are discussed. PMID- 12080369 TI - Evidence for cytoplasmic inheritance of a developmental organizer affecting growth habit and leaf shape in Antirrhinum majus. AB - A cross between two distinct, true-breeding plants of Antirrhinum majus L. showed an unexpected pattern of inheritance of growth habit in the F2, which was extended to both growth habit and leaf shape in the F3 generation of all the plants traced further. All the F3 families, offspring of individual F2 plants, were very uniform for both growth habit and leaf shape traits but differed distinctly from each other in these respects. The backcrosses of selected F3 and F4 families to the original parents in the cross did not segregate for the distinctive family phenotypes. This led to the postulate that a cytoplasmic factor was involved in the regulation and/or integration of genetic information concerned with growth habit/leaf shape. The similarity of the reciprocal backcrosses of the F3 and F4 families led to the further postulate that the proposed cytoplasmic factor was specified by both the maternal and paternal parents to a similar degree. That the gene component was segregating normally was shown by the inheritance of four marker genes for flower colour, colour pattern and flower shape. PMID- 12080370 TI - Molecular characterisation of sweet cherry (Prunus avium L.) genotypes using peach [Prunus persica (L.) Batsch] SSR sequences. AB - A total of 76 sweet cherry genotypes were screened with 34 microsatellite primer pairs previously developed in peach. Amplification of SSR loci was obtained for 24 of the microsatellite primer pairs, and 14 of them produced polymorphic amplification patterns. On the basis of polymorphism and quality of amplification, a set of nine primer pairs and the resulting 27 informative alleles were used to identify 72 genotype profiles. Of these, 68 correspond to unique cultivar genotypes, and the remaining four correspond to three cultivars that could not be differentiated from the two original genotypes of which they are mutants, and two very closely related cultivars. The mean number of alleles per locus was 3.7 while the mean heterozygosity over the nine polymorphic loci averaged 0.49. The results demonstrate the usefulness of cross-species transferability of microsatellite sequences allowing the discrimination of different genotypes of a fruit tree species with sequences developed in other species of the same genus. UPGMA cluster analysis of the similarity data divided the ancient genotypes studied into two fairly well-defined groups that reflect their geographic origin, one with genotypes originating in southern Europe and the other with the genotypes from northern Europe and North America. PMID- 12080371 TI - Genetic differentiation within and between populations of a hermaphroditic freshwater planarian. AB - Dispersal of individuals is an important factor that can influence genetic differentiation between populations. The hermaphroditic freshwater planarian Schmidtea polychroainhabits shallow regions of lakes and streams, in which they appear to be continuously distributed. In the present study we used three highly polymorphic markers for analysing small-scale and large-scale genetic structure within one, and between four natural lake populations. Genetic differentiation could already be observed between samples collected at least 13 m apart, but not between neighbouring samples, and was most pronounced between samples from different lakes. Probably due to the high variance in F(ST)values, a significant correlation between genetic differentiation and geographic distance could not be observed. These results show that individual dispersal of S. polychroa is limited, but that there is gene flow between subpopulations from the same lake. They further suggest that long-distance dispersal and gene flow between lakes, if present, is not a common process in S. polychroa. PMID- 12080372 TI - Effect of low stressful temperature on genetic variation of five quantitative traits in Drosophila melanogaster. AB - A half-sib analysis was used to investigate genetic variation for three morphological traits (thorax length, wing length and sternopleural bristle number) and two life-history traits (developmental time and larva-to-adult viability) in Drosophila melanogaster reared at a standard (25 degrees C) and a low stressful (13 degrees C) temperature. Both phenotypic and environmental variation showed a significant increase under stressful conditions in all traits. For estimates of genetic variation, no statistically significant differences were found between the two environments. Narrow heritabilities tended to be higher at 13 degrees C for sternopleural bristle number and viability and at 25 degrees C for wing length and developmental time, whereas thorax length did not show any trend. However, the pattern of genetic variances and evolvability indices (coefficient of genetic variation and evolvability), considered in the context of literature evidence, indicated the possibility of an increase in additive genetic variation for the morphological traits and viability and in nonadditive genetic variation for developmental time. The data suggest that the effect of stressful temperature may be trait-specific and this warns against generalizations about the behaviour of genetic variation under extreme conditions. PMID- 12080377 TI - Generating p53-specific cytotoxic T lymphocytes by recombinant adenoviral vector based vaccination in mice, but not man. AB - Mutations and aberrant expression of the p53 tumor suppressor protein are the most frequent molecular alterations in human malignancy. Peptides derived from the wild-type (wt) p53 protein and presented by major histocompatibility complex (MHC) molecules for T lymphocyte recognition are believed to serve as universal tumor-associated antigens for cancer immunotherapy. We studied the immunogeneicity of a recombinant replication-defective adenoviral vector encoding human full-length wt p53 (rAd/hup53) in human leukocyte antigen (HLA)-A2K(b) transgenic (Tg) mice and man. The generation of p53 epitope-specific cytotoxic T lymphocytes (CTLs) in p53-proficient and p53-deficient A2K(b)-Tg mice was affected by self-tolerance and a selective inability of rAd/hup53 to induce p53.264-272 peptide-reactive effector cells. To extend this study into a pilot clinical trial, six advanced-stage cancer patients received sequential injections of rAd/hup53. The treatment was well tolerated. To date, no evidence for objective tumor responses was observed. An amplification of humoral and cellular anti-adenoviral immune responses was demonstrated in all patients following rAd/hup53 vaccination. However, p53-reactive antibodies and HLA-A*0201 (A2.1) restricted CTLs specific for wt p53 epitopes were not generated. Tailoring p53 based cancer immunotherapy thus requires the interference with p53-specific self tolerance and the induction of the entire repertoire of p53-reactive T lymphocytes. PMID- 12080378 TI - The potential of 5-fluorocytosine/cytosine deaminase enzyme prodrug gene therapy in an intrahepatic colon cancer model. AB - Colorectal cancer can metastasize to the liver, but remain liver confined for years. A critical step in developing treatments for intrahepatic cancer involves assessment in an orthotopic intrahepatic model. The purpose of this study was to develop a noninvasive intrahepatic tumor model to study the efficacy of 5 flucytosine/yeast cytosine deaminase (5FC/yCD)-based gene therapy for liver tumors. Luciferase expressing human colorectal carcinoma (HT-29luc) cells were generated by retroviral infection and implanted in the left liver lobe of nude mice. The bioluminescence was measured every week for a period of 1 month, then animals were killed and tumors were measured by calipers. After we found a correlation between photon counts and tumor size, animals were implanted with tumors composed of either 0%, 10%, or 100% yCD/HT-29luc cells, and treated with 5FC. Tumor bioluminescence was measured during treatment and tumor histology examined at the time of death. We found that 5FC caused significant regression of yCD expressing tumors. Furthermore, visible tumors at the time of death, which emitted little bioluminescence, contained little or no viable tumor. We then developed an adenoviral vector for yCD. Intraperitoneal administration of adenovirus containing yCD led to the production of yCD enzyme within intrahepatic tumors. These results suggest that (1) intrahepatic cancer responds to 5FC when cells express yCD; (2) the luciferin-luciferase system permits non-invasive real time imaging of viable intrahepatic cancer; and (3) this system can be used to carry out gene therapy experiments using yCD adenovirus. PMID- 12080379 TI - Lipoplex-induced hemagglutination: potential involvement in intravenous gene delivery. AB - We report a study aiming to characterize the interaction of blood and blood components with lipoplexes under conditions relevant to in vivo intravenous transfection. In this study we focus on the interaction of lipoplexes with red blood cells (RBC). It was found that no significant hemolysis occurred during several hours' incubation using lipoplex compositions and lipoplex/red blood cell ratios in the range commonly used for in vivo transfection. However, the interaction of RBC with lipoplexes resulted in massive agglutination, which occurs irrespective of the type of cationic lipid or helper lipid. Agglutination was also induced by polyplexes (such as dendrimer/DNA complexes) and lipoplexes in the presence of spermidine or protamine sulfate (the latter induced hemagglutination by itself). DSPE-PEG(2000) inserted into the lipoplexes inhibits hemagglutination somewhat. In order to understand the effect of serum on the agglutination better, plasma was separated into its high molecular weight components (HMWC, >14 kDa) and its low molecular weight components (LMWC, < or = 14 kDa). These fractions were characterized for their level of proteins, primary amino groups, osmotic pressure, and electrical conductivity, and compared with saline (0.15 M NaCl). It was found that both LMWC and HMWC inhibit agglutination by themselves, although whole serum demonstrates better hemagglutination inhibition than each fraction separately. The inhibitory effect of the serum (or plasma) is explained by its effect on the electrostatics of the lipoplexes, reducing their positive charge, as was demonstrated using fluorescein phosphatidylethanolamine-labeled lipoplexes. The effect of LMWC was related to ionic strength and was equal to the effect of 0.15 M NaCl. The level of agglutination was reduced with increasing lipoplex DNA(-)/cationic lipid(+) (DNA( )/L(+)) ratio. However, at the low DNA(-)/L(+) ratio needed to achieve significant in vivo transfection after i.v. administration, massive agglutination occurred. These data suggest that i.v. administration of lipoplexes and polyplexes may lead to RBC agglutination, and the agglutinates formed may explain the localization of lipoplexes and expression of their transgenes in the lungs. PMID- 12080380 TI - Intraperitoneal gene delivery mediated by a novel cationic liposome in a peritoneal disseminated ovarian cancer model. AB - We have previously synthesized a new cationic liposome that displays high efficiency and low toxicity, 3 beta[l-ornithinamide-carbamoyl] cholesterol (O Chol), using solid-phase synthesis. In this study, O-Chol was applied to in vitro and in vivo models of ovarian cancer. Intraperitoneal gene delivery for peritoneal disseminated ovarian cancer in nude mice was achieved using a stable chloramphenicol acetyl transferase (CAT)-expressing ovarian cancer cell line (OV CA-2774/CAT), which allowed us to quantify the exact tumor burden of organs. When luciferase and beta-galactosidase genes were used as reporter genes, O-Chol showed better efficiency than other commercial transfection reagents such as lipofectin, lipofectAMINE, DC-Chol, and FuGENE 6, both in vitro and in vivo. Moreover, the transfection efficiency of this new cationic lipid reagent remained high in serum-containing medium and under serum-free conditions. Furthermore, in vivo transfection with O-Chol showed high levels of gene expression specific to peritoneal tumor cells. Consequently, the O-Chol:DNA lipoplex appears to offer potential advantages over other commercial transfection reagents because of (1) its higher level of gene expression in vitro and in vivo; (2) its reduced susceptibility to serum inhibition; and (3) its highly selective transfection into tumor cells. These results suggest that the O-Chol:DNA lipoplex is a promising tool in gene therapy for patients with peritoneal disseminated ovarian cancer. PMID- 12080381 TI - Differential effects of angiostatin, endostatin and interferon-alpha(1) gene transfer on in vivo growth of human breast cancer cells. AB - The administration of different angiogenesis inhibitors by gene transfer has been shown to result in inhibition of tumor growth in animal tumor models, but the potency of these genes has been only partially evaluated in comparative studies to date. To identify the most effective anti-angiogenic molecule for delivery by retroviral vectors, we investigated the effects of angiostatin, endostatin and interferon(IFN)-alpha(1) gene transfer in in vivo models of breast cancer induced neovascularization and tumor growth. Moloney leukemia virus-based retroviral vectors for expression of murine angiostatin, endostatin and IFN-alpha(1) were generated, characterized, and used to transduce human breast cancer cell lines (MCF7 and MDA-MB435). Secretion of the recombinant proteins was confirmed by biological and Western blotting assays. Their production did not impair in vitro growth of these breast cancer cells nor their viability, and did not interfere with the expression of angiogenic factors. However, primary endothelial cell proliferation and migration in vitro were inhibited by supernatants of the transduced cells containing angiostatin, endostatin, and IFN-alpha(1). Stable gene transfer of the IFN-alpha(1) cDNA by retroviral vectors in both MCF7 and MDA MB435 cells resulted in a marked and long-lasting inhibition of tumor growth in nude mice that was associated with reduced vascularization. Endostatin reduced the in vivo growth of MDA-MB435, but not MCF7 cells, despite similar levels of in vivo production, and angiostatin did not impair the in vivo growth of either cell line. These findings indicate heterogeneity in the therapeutic efficacy of angiostatic molecules delivered by viral vectors and suggest that gene therapy with IFN-alpha(1) and endostatin might be useful for treatment of breast cancer. PMID- 12080382 TI - HER2 peptide-specific CD8(+) T cells are proportionally detectable long after multiple DNA vaccinations. AB - We prepared a plasmid encoding 147 amino acid residues from the N terminus of c erbB-2/HER2/neu (HER2), which included both a cytotoxic T lymphocyte (CTL) epitope (HER2p63) and a helper epitope (HER2p1), using the mammalian expression vector pCAGGS-New (pCAGGS147HER2). In a parallel analysis with a Tetramer assay and CTL assay, good specificity and sensitivity of a quantitative enzyme-linked immunospot (ELISPOT) assay to detect functional HER2p63-specific CD8(+) T cells were demonstrated after intramuscular immunization of pCAGGS147HER2. In an ELISPOT assay for HER2p63, spots of IFN gamma-producing cells were first detected 10 days after the first immunization, and additional immunizations increased the number of spots. HER2p63-specific CD8(+) T cells were detected over a period of more than 10 months after the last immunization. In hosts receiving more than three immunizations, surprisingly high numbers of specific CD8(+) T cells were persistently detectable. HER2 protein-specific antibodies of IgG class with dominance of IgG2a remain detectable 6 months after single or multiple immunizations. The antibodies however, were not reactive with cell surface HER2 antigens. Total suppression of tumor growth was observed when syngeneic HER2(+) tumor cells (2 x 10(6)) were injected subcutaneously 14 days after a single immunization with pCAGGS147HER2. Furthermore, the number of pulmonary metastases decreased significantly when DNA vaccination was initiated on the day of, or 3 days after, intravenous injection (1 x 10(6) cells). PMID- 12080383 TI - Lentiviral transduction of human T-lymphocytes with a RANTES intrakine inhibits human immunodeficiency virus type 1 infection. AB - Intrakines, modified intracellular chemokines, offer a novel strategy to prevent cellular entry of HIV-1 by blocking the surface expression of HIV-1 co-receptors. To investigate potential clinical applications of the RANTES-intrakine, we explored the use of HIV-1-based lentiviral vectors for therapeutic gene transfer into T-lymphocytes. RANTES-intrakine genes can be efficiently transduced into primary human T-lymphocytes by lentiviral vectors, especially when human T lymphocytes were stimulated with CD3 and CD28 antibodies. The transduced T cells showed decreased surface expression of the chemokine receptor CCR-5, as well as CCR-1 and CCR-3. This lentivirus-mediated approach to intrakine gene transfer protected human T-lymphocytes from infection by a variety of R5-tropic HIV-1 strains. A quantitative real-time PCR assay, developed to monitor cells for HIV entry and persistence, revealed persistent low copy numbers of proviral HIV DNA in RANTES intrakine-transduced T-lymphocytes during 3-week culture, suggesting that viruses produced from infected untransduced cell populations were unable to infect the surrounding transduced T-lymphocytes. We conclude that targeting HIV-1 co-receptors to block virus entry with lentiviral vectors is an attractive approach to the control of HIV-1 infection. PMID- 12080384 TI - Intratumoral administration of an adenovirus expressing a kinase dead form of ErbB-2 inhibits tumor growth. AB - ErbB-2 is amplified or overexpressed in a number of different cancers including breast, ovarian, lung, prostate and stomach. This overexpression leads to enhanced receptor dimer formation and stabilization allowing the receptor to remain in an active state. The clinical consequences of ErbB-2 overexpression include increased tumor aggressiveness, poor prognosis, decreased patient survival and resistance to chemotherapy. As a result, a variety of different strategies are being examined to inhibit its function or expression. In this study, we explored the efficacy of a type 5 recombinant adenovirus encoding a kinase dead form of ErbB-2, AderbB-2 Delta tk, as a potential therapeutic agent for breast cancer using a murine breast model expressing constitutively active ErbB-2. Co-expression in tumor cells of the kinase dead form of ErbB-2 inhibits receptor activity and induces the death of cells expressing constitutively active ErbB-2. In addition, AderbB-2 Delta tk exhibits antitumor activity in both immune competent and immune-deficient animals with increased antitumor activity in the immune-competent animals. The results suggest both immune and non-immune mechanisms contribute to the antitumor efficacy of this vector. PMID- 12080385 TI - USH3A transcripts encode clarin-1, a four-transmembrane-domain protein with a possible role in sensory synapses. AB - Usher syndrome type 3 (USH3) is an autosomal recessive disorder characterised by the association of post-lingual progressive hearing loss, progressive visual loss due to retinitis pigmentosa and variable presence of vestibular dysfunction. Because the previously defined transcripts do not account for all USH3 cases, we performed further analysis and revealed the presence of additional exons embedded in longer human and mouse USH3A transcripts and three novel USH3A mutations. Expression of Ush3a transcripts was localised by whole mount in situ hybridisation to cochlear hair cells and spiral ganglion cells. The full length USH3A transcript encodes clarin-1, a four-transmembrane-domain protein, which defines a novel vertebrate-specific family of three paralogues. Limited sequence homology to stargazin, a cerebellar synapse four-transmembrane-domain protein, suggests a role for clarin-1 in hair cell and photoreceptor cell synapses, as well as a common pathophysiological pathway for different Usher syndromes. PMID- 12080386 TI - The elastin gene is disrupted in a family with a balanced translocation t(7;16)(q11.23;q13) associated with a variable expression of the Williams-Beuren syndrome. AB - The Williams-Beuren syndrome (WBS) is a complex developmental disorder with multisystemic manifestations including supravalvular aortic stenosis (SVAS), a so called elfin face, a hoarse voice, and a specific cognitive phenotype. Most WBS patients have a >1 Mb deletion on one of their chromosomes 7 in q11 but except for elastin, whose haploinsufficiency causes the cardiovascular malformations, it is unknown which genes in the deletion area contribute to the phenotype. We have investigated a family with a cytogenetically balanced translocation t(7;16)(q11.23;q13) in which affected individuals manifested a broad spectrum of clinical phenotypes ranging from a hoarse voice as the only feature to the full WBS phenotype. Molecular cytogenetic and DNA sequence analyses of the translocation breakpoint showed that the cytogenetic rearrangement disrupts the elastin gene locus within intron 5 in the exact same manner in all translocation carriers. The recently described large inversion of the 7q11.23 region was not present in this family. Our data demonstrate that disruption of the elastin gene by a translocation breakpoint may cause classical WBS, atypical WBS, SVAS, or no recognisable phenotype, and provide a clear example for extensive phenotypic variability associated with a position effect in humans. PMID- 12080387 TI - Chromosome analysis of epididymal and testicular spermatozoa in patients with azoospermia. AB - Azoospermic patients can now father children once spermatozoa have been retrieved from the epididymis or the testis. However, there are concerns about the risk of chromosomal abnormalities since an increase in sperm aneuploidy rate has been reported in samples from patients with abnormal sperm parameters. The purpose of this study was therefore to evaluate the sperm aneuploidy and diploidy rates for chromosomes 8, 12, 18, X and Y in spermatozoa extracted from the epididymes (n=10) or the testes (n=6) of patients with azoospermia. Ejaculated spermatozoa of healthy men (n=14) served as control. Epididymal and testicular spermatozoa had an aneuploidy rate significantly higher than that found in ejaculated spermatozoa. The aneuploidy and diploidy rates of testicular spermatozoa were higher, but not significantly different, than those found in epididymal spermatozoa. This study has shown that azoospermic patients have an increased sperm aneuploidy rate. They should therefore be given appropriate genetic counselling before entering in-vitro fertilisation programs. PMID- 12080388 TI - Segregation analysis of HDL cholesterol in the NHLBI Family Heart Study and in Utah pedigrees. AB - We investigated the genetic determination of high-density-lipoprotein cholesterol (HDL-C) levels in the NHLBI Family Heart Study by segregation analysis. Included was a total of 3755 subjects from 560 randomly selected nuclear families and 522 families selected due to a high family risk of coronary heart disease (CHD). In the whole dataset, there was no evidence for an allele at a major gene locus responsible for HDL-C levels lower than the population mean or even for significant bimodality for low levels of HDL-C. However, we observed evidence for a recessive allele that was associated with higher HDL-C levels than average. This evidence for a recessive major gene was independent of triglyceride concentrations and was most strongly observed in families recruited for CHD. The environmental model was rejected (P=0.0027) while the codominant and recessive models were not rejected (P=0.085 and P=0.133, respectively). The dominant model was also rejected (P<0.0001). In the recessive segregation model, the means of those inferred to be homozygous for the high HDL-C allele and those without the high HDL-C allele were separated by about 25 mg/dl HDL-C (73.9+/-1.99 vs 48.2+/ 0.36 mg/dl). Because these results were unexpected, segregation was tested in a separate sample of 2013 individuals in 85 large pedigrees ascertained for early heart disease deaths, early stroke deaths, and early hypertension in Utah. Similar evidence for an allele at a major gene locus for high HDL-C was found. In summary, we did not find evidence for an allele at a major gene locus associated with low HDL-C levels segregating in pedigrees recruited for the NHLBI Family Heart Study, or in pedigrees ascertained in Utah for early CHD or related phenotypes. Instead we found some evidence for the segregation of an allele associated with high HDL-C. d PMID- 12080389 TI - Locus for susceptibility for familial capillary malformation ('port-wine stain') maps to 5q. AB - Capillary malformation (CM; 'port-wine stain'), is a common vascular malformation affecting cutaneous capillary vessels in 0.3% of newborns. Increased incidence of lesions in first-degree relatives of these patients and several reported familial cases suggest that genetic factors may play a role in the pathogenesis of CM. We report the first genome-wide linkage analysis of familial CM. In the non parametric linkage analysis, strong evidence of linkage (peak Z-score 6.72, P value 0.000136) was obtained in an interval of 69 cM between markers D5S407 and D5S2098, corresponding to 5q11-5q23. Parametric linkage analysis gave a maximum combined HLOD score of 4.84 (alpha-value 0.67) at marker D5S2044 on 5q15, and analysis using only the linked families, defined a smaller, statistically significant locus CMC1 of 23 cM (peak LOD score 7.22) between markers D5S1962 and D5S652 corresponding to 5q13-5q15. Interesting candidate genes implicated in vascular and neural development, such as MEF2C, RASA1, and THBS4, are in this locus. PMID- 12080390 TI - Linkage disequilibrium and demographic history of the isolated population of the Faroe Islands. AB - The isolated population of the Faroe Islands has a history of recent expansion after being limited to a small size for centuries. Such an isolated population may be ideal for linkage disequilibrium mapping of disease genes if linkage disequilibrium (LD) extends over large regions. Analyses of 18 markers on 12q24.3, spanning a region of 4.3 Mb (16 cM), revealed extensive LD in the Faroese population. Maximum LD was found between marker pairs separated by more than 3.8 Mb. The same region had a maximum LD of only 1.2 and 1.4 Mb respectively in two outbred Danish and British populations analysed here for comparison. The analyses of gene diversity excess at 15 unlinked microsatellite markers did not reveal any sign of a severe bottleneck to have occurred within approximately 1200 years' history of the Faroese population. The extensive LD in this population may, therefore, have arisen primarily by random genetic drift. The implications for future gene mapping studies are discussed. PMID- 12080392 TI - Non-syndromic recessive deafness in Jordan: mapping of a new locus to chromosome 9q34.3 and prevalence of DFNB1 mutations. AB - Non-syndromic recessive deafness (NSRD) is the most commonly encountered form of hereditary hearing loss. The majority of NSRD cases in the Mediterranean area are linked to the DFNB1 locus (the connexin 26 GJB2 gene). Unrelated NSRD patients issued from 68 Jordanian families, were tested for mutations of the GJB2 gene by sequencing. Sixteen per cent of the families tested were linked to the DFNB1 locus. The 35delG was the only GJB2 mutation detected in these families. One of these families, presenting with four affected members and not linked to the gene, was subjected to a genome-wide search and was found to be mapped to 9q34.3 with a multipoint lodscore of 3.9. One candidate gene in the interval, coding for the chloride intracellular channel 3, CLIC3, was tested and excluded. The identification of a new NSRD locus, DFNB33, in one Jordanian family, shows the wide genetic heterogeneity that characterizes hearing impairment and the genetic diversity in Middle-Eastern populations. PMID- 12080391 TI - C677T mutation in the 5,10-MTHFR gene and risk of Down syndrome in Italy. AB - The C677T polymorphism of the MTHFR gene has been associated to maternal risk of Down syndrome, due to the detection of an higher prevalence of the T allele among mothers of children with trisomy 21, compared to control mothers. In order to confirm this association, we studied the presence of the C677T in 64 mothers of Down syndrome children and 112 controls from central Italy. An higher incidence of the mutant T allele in controls (48.2%) than in Down syndrome children mothers (44%) was detected. These results do not support the presence of an increased risk of Down syndrome in mothers carriers of the T allele in the Italian population. PMID- 12080393 TI - A cautionary note: false homozygosity for BRCA2 6174delT mutation resulting from a single nucleotide polymorphism masking the wt allele. AB - Sequencing an amplification product of the terminal segment of BRCA2 exon 11 showed apparent homozygosity for the 6174delT mutation in two healthy sisters. Subsequent sequencing of an alternate overlapping amplicon revealed the presence of the 5972C >T polymorphism, which is within the standard upstream amplification primer. This mismatch was responsible for the failure to amplify the normal (5972T) allele in both sisters who were heterozygous for the 6174delT mutation. Though the unexpected finding of apparent homozygosity for the 6174delT mutation prompted re-evaluation of the assay, the potential for false negative results due to masking of a mutation-bearing allele by such a circumstance should be a cautionary note for the testing and also in the interpretation of the results published under such assay conditions. PMID- 12080394 TI - Reasons among older Swedish women of not participating in a food survey. AB - OBJECTIVES: To (a) examine participation rate as a function of municipality, age group and living status; and (b) investigate the main reasons for exclusion and declining as stated by the women themselves. DESIGN: Analysis of participation rate and content analysis of statements given in phone calls explaining exclusion or declination from a project in which 24 h recalls and food-diaries were used. SUBJECTS: Self-managing Swedish women (n=505) were systematically selected from a stratified random sample covering single living and cohabiting women aged 64-68, 74-78 and 84-88 y living in three municipalities. RESULTS: No significant differences were found among included and declining women when municipalities and living status was analysed, but significantly more women in the oldest group were excluded (P>0.01). Among those in their 80s living at home, the usual reasons for exclusion were illness, disability or dementia, and many lived in institutions for old people. The four most used explanations to decline participation were 'lack of time', 'tired, fragile, sick or having bad memory', 'not willing to participate in scientific studies' and 'too old and nothing to contribute'. CONCLUSIONS: The participation rate was, compared with other food surveys in the older generation, fairly good, especially among those in their 80s. However, the most active and the very ill and disabled did not participate. SPONSORSHIP: The Swedish Council for Social Research, the Swedish Council for Forestry and Agricultural Research, the Swedish Foundation for Health and Care Sciences and Allergy Research and Uppsala University. PMID- 12080395 TI - Food intake patterns and risk of coronary heart disease: a prospective cohort study examining the use of traditional scoring techniques. AB - OBJECTIVE: This study analyses the risk of coronary heart disease (CHD) associated with food intake patterns. DESIGN: A cohort study with follow-up in 1996 for first admission to hospital for a CHD diagnosis or death caused by CHD (280 cases). Three food patterns were identified from a food frequency questionnaire: (1) a predefined healthy food index; (2) a prudent diet (reflecting frequent intakes of wholemeal cereals, fruit and vegetables); and (3) a Western food pattern (reflecting frequent intakes of meat products, butter and white bread) derived by factor analysis. Both factor scores had a mean of zero and a standard deviation of 1. SETTING: Copenhagen County, Denmark. SUBJECTS: A random sample of 7316 adults participated in health examinations conducted either in 1982-1984, 1987, or 1991-1992. RESULTS: The healthy food index and the Western pattern were not associated with CHD. The prudent pattern was associated with a decreased risk of CHD (Hazard ratio (HR per score unit increase)=0.85; 95% confidence intervals (CI), 0.75, 0.96), but the association vanished (HR=1.06; 95% CI, 0.93, 1.21) after controlling for confounding. Body mass index (BMI) modified the effect of the prudent and the Western patterns on CHD risk, suggesting an inverse association between both patterns and CHD in persons with low BMI, while the risk of CHD seemed to be positively related to the prudent and the Western pattern in those with high BMIs. CONCLUSIONS: This study showed no association between dietary patterns and CHD risk, but suggests that BMI modifies the relation between diet and CHD risk. PMID- 12080396 TI - Energy requirements of urban Chinese adults with manual or sedentary occupations, determined using the doubly labeled water method. AB - OBJECTIVE: To examine total energy expenditure (TEE) in relation to occupation and reported leisure time activities in free-living Chinese adults, and to determine whether measured TEE values differ from current international dietary energy recommendations. SETTING AND SUBJECTS: Seventy three weight-maintaining adults aged 35-49 y, leading unrestricted lives in urban Beijing, with a wide variety of occupations. DESIGN AND METHODS: A cross-sectional study in which TEE was determined by doubly labeled water, body composition by deuterium oxide ((2)H(2)O) dilution, resting energy expenditure (pREE) by prediction equations, and occupational and leisure time activities by questionnaire. RESULTS: For men and women respectively, TEE averaged 12.10+/-0.32 and 9.53+/-0.23 MJ/day (P<0.001), and physical activity level (PAL=TEE/pREE) was 1.77+/-0.04 and 1.66+/ 0.02 (P<0.05). Fat-free mass (FFM) was the single best predictor of TEE (adjusted r(2)=0.71, P<0.001). Occupational category (light, moderate and heavy) further predicted TEE, independent of FFM (adjusted multiple r(2)=0.82, P<0.001). Both TEE adjusted for weight and PAL increased with occupational category. Measured TEE was slightly but significantly higher than the 1985 FAO/WHO/UNU estimates for women with light occupations, but did not differ from estimates for men with light occupations, or for adults with moderate or heavy occupations. CONCLUSION: Level of occupational activity, but not duration or type of leisure activity, significantly predicted TEE in free-living urban Chinese adults. Current energy requirement recommendations slightly underestimated the energy needs of women with light occupations but were accurate for men and women with moderate and heavy occupations. SPONSORSHIP: NIH grants DK53404 and F32-DK09747. PMID- 12080398 TI - Obesity, weight change and risks for hypertension, diabetes and hypercholesterolemia in Japanese men. AB - OBJECTIVE: To examine the effects of the degree of body mass index and weight gain as the risks for hypertension, hypercholesterolemia and diabetes in Japanese men, and to compare that to the corresponding effects in a Caucasian population. DESIGN: Prospective cohort study. SETTING: Cohort of all male employees at a company. SUBJECTS: A total of 4737 male employees followed until retirement or for 4 y (longest term 4 y). RESULTS: Increase of body mass index beyond 22 kg/m(2) related to an increased risk for hypertension even after being adjusted for possible confounding factors such as age, smoking status, alcohol intake, family history and baseline value of systolic blood pressure. The risks greatly increased in subjects with a body mass index above 27 kg/m(2) for hypertension, and 29 kg/m(2) for diabetes and hypercholesterolemia. Weight gain (more than 2 kg) was strongly related to an increased risk for hypertension and hypercholesterolemia after being adjusted for possible confounding factors. However, weight gain was not related to diabetes, and weight loss did not decrease the risks for any of the three diseases. CONCLUSIONS: Among Japanese, the degrees of body mass index associated with risks for hypertension, diabetes and hypercholesterolemia were lower than those in Caucasians. The risks for hypertension and hypercholesterolemia were strongly associated with weight gain in a Japanese male population who showed a low prevalence of severe obesity, and the risks were similar to or somewhat higher than those in a Caucasian population with a high prevalence of severe obesity. SPONSORSHIP: Health Science Research Grant by the Ministry of Health and Welfare of Japan. PMID- 12080397 TI - Clinical implications of leptin and its potential humoral regulators in long-term low-calorie diet therapy for obese humans. AB - OBJECTIVE: To address the clinical implications of leptin and to re-examine the relationship between leptin and its potential humoral regulators such as insulin, nonesterified fatty acids (NEFA) and triiodothyronine (T3) in low-calorie diet (LCD) for obese humans. DESIGN: Longitudinal study. SETTING: University and foundation hospitals. SUBJECTS: Ten obese men and 10 premenopausal obese women. INTERVENTIONS: Five men and five women took 800 kcal/day LCD and another five men and five women took 1400 kcal/day balanced deficit diet (BDD) during 4 weeks. RESULTS: Plasma leptin levels in the LCD group decreased more markedly (46.2+/ 14.6 to 13.2+/-3.6 ng/ml) than that expected for the decrement in percentage fat (39.0+/-1.7 to 35.9+/-1.7%) and body mass index (BMI; 35.4+/-2.4 to 33.1+/-2.2 kg/m(2)), while that in the BDD group did not decrease significantly (14.9+/-3.5 to 13.4+/-2.8 ng/ml). The ratio of the decrease in leptin levels to that of BMI during the first week was significantly greater than that during the following 3 weeks (39.5+/-2.7 vs 29.3+/-2.1%, P=0.017). The plasma insulin and T3 levels also fell substantially in the first week and continued to decrease during the entire course. Plasma leptin levels measured weekly in each subject were correlated well with insulin (r=0.586, P=0.0003) and T3 (r=0.785, P=0.0004). Multiple regression analyses after adjustment for the time course and BMI revealed that serum levels of T3 were independently correlated with plasma leptin levels (r=0.928, P<0.0001). The plasma NEFA level was markedly elevated during the first 2 weeks and decreased thereafter. CONCLUSIONS: A rapid fall in leptin during the first week of LCD, coordinated by insulin, T3 and NEFA, should be beneficial for responding to decreased energy intake. Inversely, in view of the powerful effect of leptin on energy dissipation, the present findings suggest the potential usefulness of leptin in combination with caloric restriction for the treatment of obesity. SPONSORSHIP: The Ministry of Education, Culture, Sports, Science and Technology of Japan and the Ministry of Health, Labour and Welfare of Japan. PMID- 12080399 TI - Reduced plasma homocysteine in obese red wine consumers: a potential contributor to reduced cardiovascular risk status. AB - BACKGROUND: Moderate alcohol consumption is associated with improved vascular risk profile and decreased mortality in the middle aged. An elevated homocysteine concentration is an independent risk factor for cardiovascular disease. OBJECTIVE: To examine the relationship between alcohol consumption and homocysteine concentrations in severely obese patients (body mass index (BMI)>35). DESIGN: A careful alcohol history was obtained from 350 (male:female 1:5) consecutive patients as part of preoperative assessment for surgical treatment of obesity. Data were obtained concerning amount, frequency, timing and type of alcohol consumption. Fasting homocysteine, serum folate and vitamin B(12) concentrations were measured. Differences between groups were assessed using Student t-test, and ANOVA. Linear regression was used to assess factors influencing homocysteine concentration. RESULTS: There is a U-shaped relationship between alcohol consumption and homocysteine concentrations, with light to moderate consumption being associated with lower concentrations. Those consuming <100 g/week (n=165) of alcohol had geometric mean (95% CI of mean) serum homocysteine concentrations of 8.5 (8.2-8.9) micromol/l compared with 9.5 (9.1 9.9) micromol/l for non or rare consumers (n=153; P=0.001). The lower concentrations of homocysteine in regular consumers were associated with higher folate concentrations of 9.4 (8.6-10.2) ng/ml when compared with non-consumers 7.5 (7.1-7.8) ng/ml (P=0.001). Red wine consumers (n=42) had lower fasting concentrations of homocysteine 7.8 (7.5-8.1) micromol/l compared with 153 non consumers 9.4 (9.0-9.8) micromol/l (P<0.001), 82 beer and spirit consumers 9.0 (8.4-9.7) micromol/l (P=0.005) and 73 white wine consumers 8.8 (8.2-9.4 micromol/l (P=0.013). Red wine consumption was an independent predictor for lower homocysteine concentrations. CONCLUSION: Mild to moderate alcohol consumption, especially red wine consumption, in obese subjects is associated with lower fasting homocysteine concentrations. This may reduce cardiovascular risk and help explain the 'French paradox'. PMID- 12080400 TI - Dietary and serum alpha-tocopherol, beta-carotene and retinol, and risk for colorectal cancer in male smokers. AB - OBJECTIVE: To study the association between dietary and serum antioxidant vitamins and carotenoids and risk for colorectal cancer in male smokers. DESIGN: A prospective cohort study within a randomised, double-blind, placebo-controlled trial testing supplementation with alpha-tocopherol (50 mg/day), beta-carotene (20 mg/day) or both in preventing cancer. SUBJECTS AND METHODS: Participants of the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study with complete dietary data and serum samples available from baseline. These included 26,951 middle-aged male smokers among whom 184 colorectal cancer cases were diagnosed during 8 y of follow-up. Relative risks were calculated with Cox proportional hazards models adjusting for trial supplementation, age, body mass index, serum cholesterol, cigarettes smoked per day and physical activity. RESULTS: There was no significant association between dietary vitamin C or E, alpha-or gamma tocopherol, retinol, alpha- or beta-carotene, lycopene or lutein+zeaxanthin and risk for colorectal cancer. Serum alpha-tocopherol, beta-carotene or retinol was also not associated with the risk, neither did the season when baseline blood was drawn modify the relationship between serum beta-carotene and colorectal cancer risk. CONCLUSIONS: Our data support the results from previous studies in which no association between dietary antioxidant vitamins and carotenoids and risk for colorectal cancer has been observed. Likewise, no association between baseline serum antioxidant concentrations and colorectal cancer risk was evident. SPONSORSHIP: The Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study was supported by a contract with the US National Cancer Institute (N01-CN-45165). PMID- 12080401 TI - Depression of the glycemic index by high levels of beta-glucan fiber in two functional foods tested in type 2 diabetes. AB - OBJECTIVES: To determine the extent to which beta-glucan reduces the glycemic index (GI) of oat products and whether high levels of beta-glucan impair palatability. DESIGN: The study design was an open-label, randomized cross-over study with six treatment segments. SETTING: Free-living outpatients. SUBJECTS: Sixteen volunteers with type 2 diabetes (10 men, six women, 61+/-2 y, body mass index 29+/-2 kg/m(2), HbA1c 7.4+/-0.4%) were recruited from the St Michael's Hospital diabetes clinic. INTERVENTIONS: Volunteers were given, in random order, 50 g available carbohydrate portions of: white bread; a commercial oat bran breakfast cereal (4.4 g% beta-glucan); and a prototype beta-glucan-enriched breakfast cereal and bar, both high in beta-glucan (8.1 and 6.5 g% beta-glucan, respectively) and sweetened with fructose. Capillary blood samples were taken fasting and then 30, 60, 90, 120, 150 and 180 min after the start of the meal. Palatability was recorded using two different methods. RESULTS: The glycemic indices of the prototype beta-glucan cereal (mean+/-s.e.m.; 52+/-5) and beta glucan bar (43+/-4.1) were significantly lower than the commercial oat bran breakfast cereal (86+/-6) and white bread (100; P<0.05). All foods were highly palatable and not significantly different. It was found that the GI of the test foods used in this study decreased by 4.0+/-0.2 units per gram of beta-glucan compared to our estimate of 3.8+/-0.6 for studies reported in the literature. CONCLUSION: Addition of beta-glucan predictably reduces the GI while maintaining palatability. In a 50 g carbohydrate portion each gram of beta-glucan reduces the GI by 4 units, making it a useful functional food component for reducing postprandial glycemia. SPONSORSHIP: Nestec, Switzerland. PMID- 12080402 TI - Serum lipid profiles in Japanese women and men during consumption of walnuts. AB - OBJECTIVE: To determine the serum cholesterol, apolipoproteins and LDL oxidizability in young Japanese women and men during walnut consumption and to evaluate its active principle. DESIGN: Experimental study with a randomized design. SUBJECTS: Twenty healthy women and 20 healthy men. INTERVENTIONS: Subjects were randomly assigned to consume each of two mixed natural diets for 4 weeks in a cross-over design. Reference and walnut diets were designed and the walnut diet had 12.5% of the energy derived from walnuts (44-58 g/day). RESULTS: The total cholesterol and serum apolipoprotein B concentrations, and the ratio of LDL cholesterol to HDL cholesterol was significantly lowered in women and men when fed on the walnut diet, than when on the reference diet (P28, as compared to none of the coastal boys. Other findings showed that the students had higher levels of blood cholesterol than in the south of Norway and several other countries (about 25% had total cholesterol >5.2 mmol/l), higher intake of dietary sugar and fat than the national recommendations and little intake of seawater fish with corresponding low levels of n-3 fatty acids. This did not change after 1 y of follow up. Some 10-20% reported daily smoking and 8-13% reported that they rarely or never did physical exercise. About 30% of the students presented no factors negatively related to development of cardiovascular disease. CONCLUSION: The findings demonstrate, in the majority of students at the age of 13 and 14 y, the appearance of factors contributing to development of cardiovascular disease (CHD). SPONSORSHIP: This study was supported by the program 'Medisinsk forskning i Finnmark, University of Tromso'. PMID- 12080413 TI - Pathophysiology of preeclampsia: linking placental ischemia/hypoxia with microvascular dysfunction. AB - Studies during the past decade have provided a better understanding of the potential mechanisms responsible for the pathogenesis of preeclampsia. The initiating event in preeclampsia has been postulated to be reduced uteroplacental perfusion as a result of abnormal cytotrophoblast invasion of spiral arterioles. Placental ischemia/hypoxia is thought to lead to widespread activation/dysfunction of the maternal vascular endothelium which results in enhanced formation of endothelin, thromboxane, and superoxide, increased vascular sensitivity to angiotensin II, and decreased formation of vasodilators such as nitric oxide and prostacyclin. These endothelial abnormalities, in turn, cause hypertension by impairing renal function and increasing total peripheral resistance. While recent studies support a role for cytokines and other factors such as lipid peroxides and reactive oxygen intermediates as potential mediators of endothelial dysfunction, finding the link between placental ischemia/hypoxia and maternal endothelial and vascular abnormalities remains an important area of investigation. The quantitative importance of the various endothelial and humoral factors in mediating the vasoconstriction and elevation in arterial pressure during preeclampsia has also not been completely elucidated. PMID- 12080414 TI - The physiology of endothelial xanthine oxidase: from urate catabolism to reperfusion injury to inflammatory signal transduction. AB - Xanthine oxidoreductase (XOR) is a ubiquitous metalloflavoprotein that appears in two interconvertible yet functionally distinct forms: xanthine dehydrogenase (XD), which is constitutively expressed in vivo; and xanthine oxidase (XO), which is generated by the posttranslational modification of XD, either through the reversible, incremental thiol oxidation of sulfhydryl residues on XD or the irreversible proteolytic cleavage of a segment of XD, which occurs at low oxygen tension and in the presence of several proinflammatory mediators. Functionally, both XD and XO catalyze the oxidation of purines to urate. However, whereas XD requires NAD+ as an electron acceptor for these redox reactions, thereby generating the stable product NADH, XO is unable to use NAD+ as an electron acceptor, requiring instead the reduction of molecular oxygen for this purine oxidation and generating the highly reactive superoxide free radical. Nearly 100 years of study has documented the physiologic role of XD in urate catabolism. However, the rapid, posttranslational conversion of XD to the oxidant-generating form XO provides a possible physiologic mechanism for rapid, posttranslational, oxidant-mediated signaling. XO-generated reactive oxygen species (ROS) have been implicated in various clinicopathologic entities, including ischemia/reperfusion injury and multisystem organ failure. More recently, the concept of physiologic signal transduction mediated by ROS has been proposed, and the possibility of XD to XO conversion, with subsequent ROS generation, serving as the trigger of the microvascular inflammatory response in vivo has been hypothesized. This review presents the evidence and basis for this hypothesis. PMID- 12080415 TI - Extravascular transport of fluorescently labeled albumins in the rat mesentery. AB - OBJECTIVE: Fluorescently labeled albumin is used frequently as a tracer when monitoring microvascular permeability. Several fluorescent dyes are available for labeling protein, including fluorescein isothiocyanate (FITC) and Texas Red (TR). Because differences in leakage of dye-labeled proteins have been reported, the objective of the present study was to compare the accumulation of these two tracers in interstitium and lymph after the inflammatory event of ischemia reperfusion. METHODS: Anesthetized rats were injected intravenously with FITC labeled albumin (FITC-alb) and TR-labeled albumin (TR-alb) before 30 minutes of mesenteric ischemia. Because the tracers leaked out of the microcirculation after reperfusion, accumulation in the surrounding buffer-superfused tissue, and in separate experiments, accumulation in lymph vessels, was defined as the ratio of tissue-to-plasma and lymph-to-plasma fluorescence. RESULTS: Reperfusion induced a significant increase in tissue-to-plasma fluorescence of FITC-alb; however, no increase was observed for TR-alb. In contrast, lymph-to-plasma fluorescence of TR alb tended to be greater than FITC-alb. Reperfusion-induced increases in tissue to-plasma fluorescence of TR-alb occurred only when the superfusate was replaced with mineral oil, in which case tissue-to-plasma TR-alb fluorescence tended to be greater than FITC-alb. CONCLUSIONS: Measurement of fluorescently labeled albumin leakage from mesenteric venules depends on the dye used to label the albumin and requires an assessment of losses from the extravascular measuring region. PMID- 12080416 TI - Blood flow and red blood cell deformation in nonuniform capillaries: effects of the endothelial surface layer. AB - OBJECTIVE: A theoretical model is used to examine the mechanics of red blood cell (RBC) motion in nonuniform capillaries. The model includes effects of the endothelial surface layer (ESL), which is a layer of macromolecules adjacent to the endothelium and which impedes plasma flow. METHODS: The motion of an RBC traversing a capillary with diameter varying sinusoidally between 5.4 microm and 7.4 microm is simulated numerically. The ESL is assumed to be 0.7-microm wide and deformable. Axisymmetric RBC shapes are assumed. Lubrication theory is used to analyze the motion of plasma around the RBC and through the ESL. RESULTS: In a nonuniform capillary with no ESL, moving RBCs undergo large transient deformations and predicted flow resistance is substantially higher than in a uniform capillary with the same mean diameter. The presence of a deformable ESL reduces the transient fluid shear stresses and deformations experienced by RBCs traversing a nonuniform capillary. With an ESL, the increase in flow resistance resulting from nonuniformity is less than twofold versus three- to fourfold with no ESL in vessel geometries with the same ESL-free luminal region. CONCLUSIONS: The presence of the ESL reduces the impact of capillary irregularity on flow resistance and may protect RBCs traversing irregular capillaries from damage due to large, rapidly fluctuating external stresses. PMID- 12080417 TI - Effect of the cytoskeletal fixation agent phalloidin on transcapillary albumin transport and interstitial fluid pressure in anaphylaxis in the wistar rat. AB - OBJECTIVE: Interstitial fluid pressure (P(if)) plays an important role in controlling interstitial fluid volume. In the early phase of rapid edema formation, such as in dextran-induced anaphylaxis in the Wistar rat, P(if) falls from -0.5 mm Hg to a value between -5 and -10 mm Hg. It is believed that P(if) is controlled by the interaction between connective tissue cells and the extracellular matrix. This hypothesis was tested by studying dextran-induced edema formation and the subsequent changes in P(if) in response to a pretreatment with phalloidin, a reagent that fixes the actin filaments within the cell. METHODS: P(if) was measured in anesthetized female Wistar rats by using a micropuncture technique. The rats were pretreated with phalloidin followed by dextran. Albumin extavasation (E(alb)) was measured as the extravascular space of (125)I-labeled human serum albumin (HSA) after 25 minutes. Total tissue water (TTW) was calculated as the difference between wet weight and dry weight divided by dry weight. Localization of phalloidin was determined histologically by using rhodamine-labeled phalloidin. RESULTS: Control P(if) values of -0.3 +/- 0.6 mm Hg dropped to -3.1 +/- 0.8 mm Hg after dextran treatment (p < 0.05). Pretreatment with phalloidin completely abolished this decrease in P(if), giving values of 0.6 +/- 0.2 mm Hg (p < 0.05, compared to dextran). The E(alb) in control rats of 0.02 +/- 0.02 mL/g DW increased to 1.35 +/- 0.43 mL/g DW after dextran treatment. Pretreatment with phalloidin before dextran treatment lowered the dextran value to 0.59 +/- 0.32 mL/g DW. CONCLUSION: This study shows that pretreatment with phalloidin, before the administration of dextran, abolishes the lowering of P(if) and edema formation, which is detected after i.v. injection of dextran alone. PMID- 12080418 TI - Evidence for dermal angiogenesis in breast cancer related lymphedema demonstrated using dual-site fluorescence angiography. AB - OBJECTIVE: To determine whether the skin expansion associated with breast cancer related lymphedema (BCRL) reduces microvessel density or causes a compensatory angiogenesis. METHODS: Dermal microvessel density was measured by simultaneous fluorescence video angiography in the ipsilateral and contralateral forearms of 25 patients treated for breast cancer, 12 with BCRL (excess volume 30 +/- 20%) and 13 without. RESULTS: Microvessel density was as high in the swollen arm (27.2 +/- 7.2 mm(-2)) as in the contralateral arm (24.2 +/- 6.2 mm(-2)) despite a 16 +/ 12% increase in local circumference. Total microvessels in a 1-mm annulus of skin was 30% greater in the swollen arm (752 +/- 211) than the contralateral arm (578 +/- 157; p = 0.005, t-test). Microvessel density was similar in each arm in control patients (ipsilateral 24.1 +/- 6.2 mm(-2), contralateral 26.1 +/- 8.8 mm( 2)). CONCLUSIONS: The number of microvessels increased in the expanded skin to maintain microvessel density at a normal level. Therefore, angiogenesis occurs during chronic limb swelling. The control group results indicate that angiogenesis is not caused by the cancer treatment. PMID- 12080419 TI - Use of degenerate primers and touchdown PCR to amplify a halogenase gene fragment from Streptomyces venezuelae ISP5230. AB - Consensus amino acid sequences of FADH(2)-dependent bacterial halogenases were used to design PCR primers amplifying a halogenase gene fragment from the chloramphenicol producer Streptomyces venezuelae ISP5230. The sequence-specific degenerate primers (MPF1 and MPR2) were used with a touchdown PCR procedure in the first PCR-assisted cloning of a halogenase gene fragment. In the region of the 290-bp PCR product containing the reverse primer, the deduced amino acid sequence exhibited characteristics of a beta-alpha-beta fold present in FAD binding sites of certain monooxygenases. When used to probe Southern blots of restriction-enzyme-digested DNA, the [alpha-(32)P]dCTP-labeled PCR product hybridized specifically with DNA fragments from genomic DNA of S. venezuelae ISP5230. Primers MPF1 and MPR2 also allowed amplification by PCR of approximately 290-bp DNA fragments from several other streptomycetes. The fragments from Streptomyces aureofaciens NRRL2209 and Streptomyces coelicolor A3(2) showed sequence identity with halogenase genes from these species. Thus, the PCR primers are of potential value for amplification and subsequent isolation of actinomycete halogenase genes. PMID- 12080420 TI - In vitro colonization of hydrophilic contact lenses by Aspergillus niger. AB - In vitro colonization of hydrophilic contact lenses by Aspergillus niger was investigated. Five strains of the fungus, four polymers, two culture media and four incubation periods were considered for analysis. Only the 2700 strain colonized the lenses. The degrees of adhesion and invasion varied significantly according to the characteristics of the culture under investigation. PMID- 12080421 TI - Role of dose concentration in biocide efficacy against Pseudomonas aeruginosa biofilms. AB - Pseudomonas aeruginosa entrapped in alginate gel beads to form artificial biofilms resisted killing by chlorine, glutaraldehyde, 2,2-dibromo-3 nitrilopropionamide (DBNPA), and an alkyl dimethyl benzyl ammonium compound (ADBAC). The degree of resistance was quantified by a resistance factor that compared killing times for biofilm and planktonic cells in response to the same concentration of antimicrobial agent. Resistance factors averaged 120 for chlorine, 34 for glutaraldehyde, 29 for DBNPA, and 1900 for ADBAC. In every case, resistance factors decreased with increasing concentration of the antimicrobial agent. An independent analysis of the concentration dependence of the apparent rates of killing of planktonic and biofilm bacteria showed that elevating the treatment concentration increased bacterial killing more in the biofilm than it did in a suspension culture. Calculation of a transport modulus comparing the rates of biocide reaction and diffusion suggested that at least part of the biofilm resistance to chlorine, glutaraldehdye, and DBNPA could be attributed to incomplete or slow penetration of these agents into the biofilm. Time-kill curves were nonlinear for biofilm bacteria in some cases. The shapes of these curves implicated retarded antimicrobial penetration for chlorine and glutaraldehyde and the presence of a tolerant subpopulation for DBNPA and ADBAC. The results indicate that treating biofilms with a concentrated dose of biocide is more effective than using prolonged doses of a lower concentration. PMID- 12080422 TI - Optimization of fed-batch fermentation for xylitol production by Candida tropicalis. AB - Xylitol, a functional sweetener, was produced from xylose by biological conversion using Candida tropicalis ATCC 13803. Based on a two-substrate fermentation using glucose for cell growth and xylose for xylitol production, fed batch fermentations were undertaken to increase the final xylitol concentration. The effects of xylose and xylitol on xylitol production rate were studied to determine the optimum concentrations for fed-batch fermentation. Xylose concentration in the medium (100 g l(-1)) and less than 200 g l(-1) total xylose plus xylitol concentration were determined as optimum for maximum xylitol production rate and xylitol yield. Increasing the concentrations of xylose and xylitol decreased the rate and yield of xylitol production and the specific cell growth rate, probably because of an increase in osmotic stress that would interfere with xylose transport, xylitol flux to secretion to cell metabolism. The feeding rate of xylose solution during the fed-batch mode of operation was determined by using the mass balance equations and kinetic parameters involved in the equations in order to increase final xylitol concentration without affecting xylitol and productivity. The optimized fed-batch fermentation resulted in 187 g l(-1) xylitol concentration, 0.75 g xylitol g xylose(-1) xylitol yield and 3.9 g xylitol l(-1) h(-1) volumetric productivity. PMID- 12080423 TI - Selection of microbial consortia for treating metal-working fluids. AB - The aim of this research was to determine the effectiveness of a strategy for constructing microbial consortia for treating chemically mixed industrial effluent, based on a more thorough understanding of communities within waste metal-working fluids (MWFs). Complementary phenotypic and genotypic methods revealed that the microbial communities in spent MWFs had low diversity and were very similar in species composition in samples originating from different locations and uses. Of 65 bacterial isolates studied, only 9 species were identified using fatty acid methyl ester (FAME) analysis. The results of genotypic analysis by denaturing gradient gel electrophoresis (DGGE) were congruent with observations made using FAME analysis. The metabolic potential of the isolates was assessed in terms of assimilation ability and tolerance of co contaminants. The three isolates, selected (Clavibacter michiganensis, Methylobacterium mesophilicum, and Rhodococcus erythropolis) to form a consortium, were representative of three of the four most abundant populations and when combined could utilise or tolerate all of the individual MWF components, including the biocide and the recalcitrant compound benzotriazole. PMID- 12080424 TI - Influence of Saccharomyces cerevisiae strains on fermentation and flavor compounds of white wines made from cv. Emir grown in Central Anatolia, Turkey. AB - The effect of inoculation with selected Saccharomyces cerevisiae strains was studied on fermentation and flavor compounds of wines made from Vitis vinifera L. cv. Emir grown in Central Anatolia, Turkey. Flavor compounds were analysed and identified by GC-FID and GC-MS, respectively. The total concentrations of flavor compounds did not increase with the addition of indigenous and commercial wine yeasts, but differences were noted in individual volatile compounds. Cluster and factor analyses of flavor compounds also showed that wines produced were different depending on the wine strain used. Wines were completely fermented to less than 1.4 g/l residual sugar. Yeasts other than S. cerevisiae survived longer than previously reported. Inoculation with selected strains increased the ethanol level. PMID- 12080425 TI - Modulation of gene expression from the arabinose-inducible araBAD promoter. AB - The arabinose-inducible P(BAD) promoter suffers from all-or-none gene expression in which cells harboring the natively controlled arabinose transport gene (araE) are either induced or uninduced, the relative fraction of which is controlled by the concentration of arabinose. The population-averaged variation in expression from P(BAD) as a function of inducer concentration is proportional to the percentage of cells that are fully induced (vs. uninduced) rather than the level of expression in individual cells. Because of its undesirable effects on the expression of heterologous genes, the all-or-none phenomenon was eliminated in Escherichia coli by expression of araE from arabinose-independent (either the Lactococcus lactis constitutive or IPTG-inducible lac) promoters. In these arabinose-transport engineered cells, variation in P(BAD) expression with arabinose concentration was a result of variation of the expression level in individual cells with all cells in the population having approximately the same induction level. PMID- 12080426 TI - Evaluating the potential efficacy of three antifungal sealants of duct liner and galvanized steel as used in HVAC systems. AB - Current recommendations for remediation of fiberglass duct materials contaminated with fungi specify complete removal, which can be extremely expensive, but in place duct cleaning may not provide adequate protection from regrowth of fungal contamination. Therefore, a common practice in the duct-cleaning industry is the postcleaning use of antifungal surface coatings with the implication that they may contain or limit regrowth. However, even the proper use of these products has generally been discouraged because little research has been conducted on the effectiveness of most products as used in heating, ventilating, and air conditioning (HVAC) systems. Three different coatings were evaluated on fiberglass duct liner (FGDL). Two of the three coatings were able to limit growth in the 3-month study; the third did not. One of the coatings that was able to limit growth was further evaluated in a comparison of FGDL or galvanized steel (GS) under conditions that mimicked their use in HVAC systems. The results showed that both moderately soiled and heavily soiled uncoated FGDL and GS duct material can support fungal growth, but that GS duct material was more readily cleaned. The use of an antifungal coating helped limit, but did not fully contain, regrowth on FGDL. No regrowth was detected on the coated GS. PMID- 12080427 TI - High-level production of D-mannitol with membrane cell-recycle bioreactor. AB - Ten heterofermentative lactic acid bacteria were compared in their ability to produce D-mannitol from D-fructose in a resting state. The best strain, Leuconostoc mesenteroides ATCC-9135, was examined in high cell density membrane cell-recycle cultures. High volumetric mannitol productivity (26.2 g l(-1) h(-1)) and mannitol yield (97 mol%) were achieved. Using the same initial biomass, a stable high-level production of mannitol was maintained for 14 successive bioconversion batches. Applying response surface methodology, the temperature and pH were studied with respect to specific mannitol productivity and yield. Moreover, increasing the initial fructose concentration from 100 to 120 and 140 g l(-1) resulted in decreased productivities due to both substrate and end-product inhibition of the key enzyme, mannitol dehydrogenase (MDH). Nitrogen gas flushing of the bioconversion media was unnecessary, since it did not change the essential process parameters. PMID- 12080428 TI - Mecamylamine (Inversine): an old antihypertensive with new research directions. AB - Mecamylamine (Inversine), the first orally available antihypertensive agent, is now rarely used. Although celebrated in the 1950s, mecamylamine fell out of favour because of its widespread ganglionic side effects at antihypertensive doses (30-90 mg/day). However, recent studies suggest that mecamylamine is very effective at relatively low doses (2.5-5 mg b.i.d.) for blocking the physiological effects of nicotine and improving abstinence rates in smoking cessation studies, particularly for women. When these lower doses of mecamylamine are given, patients do not experience the severity of side effects that made the drug unpopular for the treatment of hypertension. Tobacco smoking is a strong risk factor for cardiovascular morbidity, including accelerated atherosclerosis and increased risk of heart attacks. Though currently untested, the available evidence suggests that low-dose mecamylamine therapy might reduce blood pressure variability and atherogenetic lipid profile in smokers. With this in mind, mecamylamine should be an important research tool in the field of hypertension research, particularly in recalcitrant smokers with mild to moderate hypertension. PMID- 12080429 TI - Participation of renal and circulating endothelin in salt-sensitive essential hypertension. AB - Salt sensitivity of blood pressure is a cardiovascular risk factor, independent of and in addition to hypertension. In essential hypertension, a conglomerate of clinical and biochemical characteristics defines a salt-sensitive phenotype. Despite extensive research on multiple natriuretic and antinatriuretic systems, there is no definitive answer yet about the major causes of salt-sensitivity, probably reflecting the complexity of salt-balance regulation. The endothelins, ubiquitous peptides first described as potent vasoconstrictors, also have vasodilator, natriuretic and antinatriuretic actions, depending on their site of generation and binding to different receptors. We review the available data on endothelin in salt-sensitive essential hypertension and conclude that abnormalities of renal endothelin may play a primary role. More importantly, the salt-sensitive patient may have blood pressure-dependency on endothelin in all states of salt balance, thus predicting that endothelin receptor blockers will have a major therapeutic role in salt-sensitive essential hypertension. PMID- 12080430 TI - The role of home blood pressure measurement in managing hypertension: an evidence based review. AB - This article reviews the current literature regarding the role of home measurement of blood pressure (BP) in the management of hypertension. Subjects with hypertension can use simple automated devices to measure their own BP at home. The results can be accurate and reliable, and because multiple readings allow a mean value to be calculated, a better estimate of the underlying BP level is obtained. Home measurement of BP gives results which are equivalent to the accepted 'gold standard' measure of ambulatory BP values, whilst using a simpler and much less expensive technique which is therefore more widely available. Both methods are better than conventional office measurements in identifying the 'true' or underlying mean BP level and identifying falsely raised levels or 'white coat hypertension'. White coat hypertension confounds the treatment of hypertension, but may not be entirely harmless. Ambulatory BP is a better predictor of cardiovascular prognosis than clinic BP. The use of home BP measurement as an equivalent, feasible, and (apparently) more cost-effective technique to measure BP in hypertension, should enable groups of patients with a poorer prognosis to be identified and their treatment adjusted in order to improve their prognosis. PMID- 12080431 TI - Blood pressure and urinary excretion of electrolytes in Spanish schoolchildren. AB - Despite the importance of hypertension in adults, its effects on child health are poorly understood. This cross-sectional epidemiological study was designed to look for a relationship between elevated blood pressure (BP) in children and 24-h urinary excretion of sodium (Na) and potassium (K), and between BP and dietary salt intake. The study population was all 59 856 schoolchildren aged 6 to 14 years in the province of Almeria in southern Spain, among whom 613 participants were chosen randomly for study. We measured 24-h urinary Na and K concentrations, systolic and diastolic BP, body weight and height. There was a weak correlation between Na excretion and systolic BP (r = 0.18, 95% confidence interval 0.10 0.26), and between K excretion and systolic BP (r = 0.49, 95% CI = 0.04-0.20). Body weight was the variable that best correlated with systolic (r = 0.49, 95% CI = 0.43-0.55) and diastolic BP, and with Na excretion (r = 0.48, 95% CI = 0.42 0.55). Multiple regression analysis also showed that body weight was the variable that best correlated with systolic BP (b = 0.58), although the variables in the equation explained little of the total variability in BP (26%). These correlations were significant at P < 0.05. In conclusion urinary electrolytes correlated poorly with BP in a sample of Spanish schoolchildren. Body weight was the only variable that showed a weak relationship with BP and Na excretion. PMID- 12080433 TI - Exacerbation of insulin resistance and postprandial triglyceride response in newly diagnosed hypertensive patients with hypertriglyceridaemia. AB - The purpose of the study is to examine the differences in insulin resistance and postprandial triglyceride (TG) response between hypertensive patients with or without hypertriglyceridaemia. The study is a comparative cohort study with matching. Thirty-one newly diagnosed hypertensive patients without any medication were recruited from a health survey. The participants were further divided into two groups: those with fasting TG <2.26 mmol/L, and those with TG between 2.26 and 5.65 mmol/L. Both groups were matched in age, sex, body mass index and waist circumference. Each patient received a 75-g oral glucose tolerance test, an insulin suppression test, and a 1000 kcal high fat mixed meal test. The hypertriglyceridaemic hypertensive patients had significantly higher fasting insulin, 2-h plasma glucose, 2-h insulin, and steady-state plasma glucose (SSPG) (13.16 +/- 1.87 vs 9.76 +/- 3.18 mmol/L). They also had a greater postprandial TG response to the challenge of mixed meal (DeltaAUC 20.76 +/- 10.06 vs 7.97 +/- 3.18 mmol 8 h/L). The postprandial TG response was closely correlated (r = 0.72 0.95, P < 0.0001) with fasting TG in all hypertensive patients. Both fasting TG levels and postprandial TG response were significantly (P < 0.05) correlated with SSPG. In conclusion, the hypertensive patients with hypertriglyceridaemia were more insulin resistant than those without it. Exacerbation of postprandial hypertriglyceridaemia was identified in these patients. The TG response to the challenge of high fat meal was significantly correlated with fasting TG and insulin resistant in them. The results provide a rationale for the alleviation of insulin resistance and hypertriglyceridaemia in these atherosclerosis-prone hypertensive patients. PMID- 12080432 TI - Genetic differences in ethanol metabolizing enzymes and blood pressure in Japanese alcohol consumers. AB - Orientals have unique genetic polymorphisms in ethanol metabolizing enzymes, such as alcohol dehydrogenase-2 (ADH2), aldehyde dehydrogenase-2 (ALDH2) and cytochrome P450-2E1 (CYP2E1). Of the three studies conducted to clarify the influence of ALDH2 genotypes on sensitivity to the pressor effects of alcohol in Japanese, only one was suggested, though indirectly, higher sensitivity in drinkers having the genotype of inactive ALDH2. This discrepancy prompted us to determine ADH2, ALDH2 and CYP2E1 genotypes in the genomic DNA extracted from white blood cells of 855 healthy middle-aged Japanese men, and to analyse the associations with the alcohol-blood pressure (BP) relationship. No marked differences were found in the relationship among the genotypes of ALDH2, although the subjects with intact ALDH2 showed a slightly higher BP than those with inactive ALDH2 probably due to under-reporting of alcohol consumption in those with intact ALDH2 who could thus drink more. No significant influence of ADH2 genotypes was observed. A higher BP was noted in large volume alcohol consumers having c2/c2 genotype of CYP2E1. Multivariate regression analysis adjusting for the effects of age, body mass index and the volume of alcohol consumed, all of which are strong determinants of BP levels, showed only a marginal effect of c2 allele of CYP2E1 on diastolic BP elevations with increases in alcohol consumption. Thus it is concluded that the genetic polymorphisms in ethanol metabolizing enzymes do not greatly influence the alcohol-BP relationship in Japanese men. PMID- 12080434 TI - Hypertension and clusters of risk factors in different stroke subtypes (an analysis of Hungarian patients via Budapest Stroke Data Bank). AB - Our aim was to determine the frequency of hypertension in the Hungarian stroke population, and to compare it with the data of other stroke registries. We attempted to find characteristic cluster-like associations between hypertension and another nine risk factors in different stroke subtypes and to ascertain the role of hypertension in leukoaraiosis, in early mortality, in stroke recurrence and in the case fatality rate up to 10 years. Risk factor profile of 500 unselected acute stroke cases of the Budapest Stroke Data Bank were analysed. We compared data of hypertensive stroke patients to those of unaffected ones. LIFEREG procedure of the SAS software package, cluster analysis, logistic regression, Pearson's correlation coefficient and Student's t-test were used as statistical methods. Hypertension was documented in 75% of the patients. The largest clusters were formed by the following groups: atherosclerotic stroke, hypertension with ischaemic heart disease; lacunar stroke and haemorrhage, hypertension with elevated serum cholesterol, cardiogenic embolism, ischaemic heart disease with atrial fibrillation. The case fatality rate was significantly higher in the group of hypertensive patients with ischaemic heart disease relative to those without it during the 10 years follow-up period. In the leukoaraiosis-group, systolic blood pressure was significantly higher than in the non-leukoaraiosis group. The rate of hypertension was higher than in other stroke registries. Hypertension appears to be the most frequent risk factor in stroke patients but case fatality rate is determined by presence or absence of ischaemic heart disease. PMID- 12080435 TI - Renovascular disease in patients with hypertension: detection with duplex ultrasound. AB - The aim of this study was to evaluate the accuracy of duplex ultrasound for the diagnosis of renovascular disease in a cohort of hypertensive patients. In 78 patients suspected of renovascular hypertension on clinical grounds duplex ultrasound examination of the renal arteries was performed. Renal angiography was used as the standard of reference. Duplex ultrasound was inconclusive in 11 kidneys (7%). None of the supernumerary renal arteries was detected with duplex ultrasound. The overall prevalence of significant renovascular disease (> or =50% stenosis) was 20%. Based on the combination of parameters at thresholds commonly applied in current literature: ie PSV(max) >180 cm/sec and RAR >3.5 the overall sensitivity of duplex ultrasound for detection of haemodynamically significant renovascular disease was 50.0% with a specificity of 91.3% (PPV: 87.9%; NPV: 59.1). Lowering the thresholds for both parameters improved the test results at the cost of a significant increase of false positive examinations. In a population of hypertensive patients clinically suspected of renovascular hypertension, only limited results for duplex ultrasound could be acquired in the detection of renovascular disease. This result, in combination with the wide range of sensitivities and specificities published in international literature and the relatively large number of incomplete examinations does not support the general application of duplex ultrasound as a screening procedure for detection and assessment of renovascular disease. PMID- 12080436 TI - The effect of hormone replacement therapy on arterial blood pressure and vascular compliance in postmenopausal women with arterial hypertension. AB - Arterial pathology is a major contributor to cardiovascular disease, morbidity and mortality. Women are at higher risk of cardiovascular disease after menopause. Arterial stiffness determined by pulse wave velocity, increases with age both in men and women, whereas arterial compliance in premenopausal women is greater than in men of similar age. This difference is lost in the postmenopausal years, with evidence of rapid decline in arterial compliance in the perimenopausal period. Loss of hormonal modulation is a likely explanation for reduced arterial compliance in postmenopausal women. Long-term treatment with hormone replacement therapy (HRT) may be expected to partially reverse the increase in arterial stiffness. The aim of the study was to analyse the effect of HRT on blood pressure and arterial compliance in postmenopausal women with arterial hypertension receiving hypotensive drugs. The results in the present study of postmenopausal women with mild to moderate arterial hypertension receiving HRT showed only a transient tendency towards lower blood pressure. In our study HRT was found to improve arterial compliance at 3 months after HRT, and the effect was maintained throughout 12 months. The increased arterial compliance in women receiving HRT was independent of blood pressure. In parallel with decreasing pulse wave velocity women receiving HRT had lower total and low density lipoprotein cholesterol. The conclusions were that after 1 year HRT in postmenopausal women with arterial hypertension improves circadian blood pressure pattern, but it does not affect significantly blood pressure values and variability. The present study also shows that HRT significantly inhibits age related rigidity of large arteries. PMID- 12080437 TI - The effect of the combination of Mediterranean diet and leisure time physical activity on the risk of developing acute coronary syndromes, in hypertensive subjects. AB - The World Health Organization reports that the number of hypertensives, worldwide, is estimated to be 600 million people. In addition a considerable proportion of hypertensive subjects remains untreated or uncontrolled. In this work we investigated the combined effect of physical activity and Mediterranean diet on coronary risk, in hypertensives. Thus we randomly selected, from all Greek regions, 848 hospitalised patients (695 males, 58 +/- 10 years old and 153 females, 65 +/- 9 years old) with a first event of coronary heart disease (CHD) and 1078 paired, by sex, age, region controls, without any suspicions for CHD. Physically active were those who reported non-occupational physical activity more than once per week. Subjects 'closer' to the Mediterranean diet were assessed through a special nutrient questionnaire. A total of 418 (49%) of the patients and 303 (28%) of the controls were hypertensive. Of these, 115 (27%) patients and 70 (23%) controls were untreated, 148 (35%)-111 (36%) were uncontrolled and 155 (38%)-122 (41%) were controlled (P-value <0.01). One hundred and sixty-two (19%) of the patients and 265 (25%) of the controls (P < 0.01) were 'closer' to the combination of Mediterranean type of diet and physical activity. The analysis showed that the previous combination is related to a 25% reduction of the coronary risk in controlled hypertensive subjects (OR = 0.75, P < 0.01), a 11% reduction in untreated (OR = 0.89, P < 0.05) and 17% reduction (OR = 0.83, P < 0.05) in uncontrolled, after adjusting for age, sex, educational and financial level and the conventional cardiovascular risk factors. Consequently, the adoption of Mediterranean diet by physically active subjects seems to reduce significantly the coronary risk and prevent, approximately, the one-third of acute CHD, in controlled hypertensive subjects. PMID- 12080438 TI - The oscillometric wrist devices: an estimate of the 'supine' error. PMID- 12080439 TI - Aldosterone excess and essential hypertension. PMID- 12080440 TI - Obesity and the polycystic ovary syndrome. AB - The polycystic ovary syndrome (PCOS) is a condition characterized by hyperandrogenism and chronic oligo-anovulation. However, many features of the metabolic syndrome are inconsistently present in the majority of women with PCOS. Approximately 50% of PCOS women are overweight or obese and most of them have the abdominal phenotype. Obesity may play a pathogenetic role in the development of the syndrome in susceptible individuals. In fact, insulin possesses true gonadotrophic function and an increased insulin availability at the level of ovarian tissue may favour excess androgen synthesis. Obesity, particularly the abdominal phenotype, may be partly responsible for insulin resistance and associated hyperinsulinemia in women with PCOS. Therefore, obesity-related hyperinsulinemia may play a key role in favouring hyperandrogenism in these women. Other factors such as increased estrogen production rate, increased activity of the opioid system and of the hypothalamic-pituitary-adrenal axis, decreased sex hormone binding globulin synthesis and, possibly, high dietary lipid intake, may be additional mechanisms by which obesity favours the development of hyperandrogenism in PCOS. Irrespective of the pathogenetic mechanism involved, obese PCOS women have more severe hyperandrogenism and related clinical features (such as hirsutism, menstrual abnormalities and anovulation) than normal-weight PCOS women. This picture tends to be more pronounced in obese PCOS women with the abdominal phenotype. Body weight loss is associated with beneficial effects on hormones, metabolism and clinical features. A further clinical and endocrinological improvement can also be achieved by adding insulin-sensitizing agents and/or antiandrogens to weight reduction programmes. These obviously emphasize the role of obesity in the pathophysiology of PCOS. PMID- 12080441 TI - Impaired ('diabetic') insulin signaling and action occur in fat cells long before glucose intolerance--is insulin resistance initiated in the adipose tissue? AB - This review postulates and presents recent evidence that insulin resistance is initiated in the adipose tissue and also suggests that the adipose tissue may play a pivotal role in the induction of insulin resistance in the muscles and the liver. Marked impairments in insulin's intracellular signaling cascade are present in fat cells from type 2 diabetic patients, including reduced IRS-1 gene and protein expression, impaired insulin-stimulated PI3-kinase and PKB/Akt activities. In contrast, upstream insulin signaling in skeletal muscle from diabetic subjects only shows modest impairments and PKB/Akt activation in vivo by insulin appears normal. However, insulin-stimulated glucose transport and glycogen synthesis are markedly reduced. Similar marked impairments in insulin signaling, including reduced IRS-1 expression, impaired insulin-stimulated PI3 kinase and PKB/Akt activities are also seen in some (approximately 30%) normoglycemic individuals with genetic predisposition for type 2 diabetes. In addition, GLUT4 expression is markedly reduced in these cells, similar to what is seen in diabetic cells. The individuals with reduced cellular expression of IRS-1 and GLUT4 are also markedly insulin resistant and exhibit several characteristics of the Insulin Resistance Syndrome.Thus, a 'diabetic' pattern is seen in the fat cells also in normoglycemic subjects and this is associated with a marked insulin resistance in vivo. It is proposed that insulin resistance and/or its effectors is initiated in fat cells and that this may secondarily encompass other target tissues for insulin, including the impaired glucose transport in the muscles. PMID- 12080442 TI - Hormonal regulation of interleukin-6 production in human adipocytes. AB - OBJECTIVE: To elucidate the hormonal regulation of interleukin-6 (IL-6) production by human adipose tissue and its relation to leptin. DESIGN: In vitro study. Human adipocytes were incubated with dexamethasone (with or without RU486), norepinephrine and epinephrine (with or without propranolol), or insulin. MEASUREMENTS: IL-6 and leptin secretion by human adipocytes. RESULTS: A gradual increase in IL-6 secretion by adipocytes during differentiation was observed. A positive correlation was found between basal IL-6 release and both glycerol 3 phosphate dehydrogenase activity--a marker of adipocyte differentiation-and leptin release. Dexamethasone decreased IL-6 secretion and increased leptin secretion in a dose-dependent manner. Both catecholamines increased IL-6 and leptin secretion. The effects of dexamethasone and catecholamines on IL-6 and leptin were abrogated by RU486 and propranolol, respectively. Incubation with insulin resulted in a dose-dependent stimulation of IL-6 and leptin secretion. CONCLUSIONS: IL-6 is produced by human adipocytes and is a potential marker of adipocyte differentiation. Furthermore it is a hormonally regulated cytokine, suppressed by glucocorticoids, and stimulated by catecholamines and insulin in physiological concentrations. PMID- 12080443 TI - Effects of Trecadrine, a beta3-adrenergic agonist, on leptin secretion, glucose and lipid metabolism in isolated rat adipocytes. AB - OBJECTIVE: Leptin, a hormone produced in adipocytes, is a key signal in the regulation of food intake and energy expenditure. Beta-adrenergic agonists have been shown to inhibit leptin gene expression and leptin secretion. The mechanisms underlying the inhibitory effects of beta-adrenergic agonists have not been established. In this study, we examined the effects of Trecadrine, a novel beta3 adrenergic agonist, on basal and insulin-stimulated leptin secretion in isolated rat adipocytes. Because insulin-stimulated glucose metabolism is an important regulator of leptin expression and secretion by the adipocytes, the effects of Trecadrine on indices of adipocyte metabolism were also examined. MEASUREMENTS: Isolated adipocytes were incubated with Trecadrine (10(-8)-10(-4) M) in the absence or presence of insulin (1.6 nM). Leptin secretion, glucose utilization, lactate production, glucose incorporation into CO(2) and triglyceride, as well as lipolysis (glycerol release) were determined. RESULTS: Trecadrine induced a concentration-dependent inhibition of basal leptin secretion. Trecadrine also decreased insulin-stimulated leptin secretion; however, the effect was not as pronounced as in the absence of insulin. Treatment of adipocytes with Trecadrine increased basal glucose utilization and produced a further increase in insulin stimulated glucose utilization. Basal lactate production was also increased by Trecadrine; however, the proportion (percentage) of glucose carbon released as lactate was unaffected. In the presence of insulin, absolute lactate production was unaffected by Trecadrine at 96 h. However, the percentage of glucose carbon released as lactate was significantly decreased by insulin treatment, and was further decreased by the co-treatment with Trecadrine. Trecadrine induced a dose dependent increase of the absolute amount of glucose incorporated into triglyceride. However, the percentage of glucose utilized that was incorporated into triglyceride was unaffected by Trecadrine. Trecadrine did not modify the proportion of glucose utilized that was oxidized to CO(2). Trecadrine increased glycerol release after 96 h of treatment. Glycerol release was negatively correlated with leptin secretion. CONCLUSIONS: These results suggest that alterations of glucose metabolism are not directly involved in the effects of beta3-adrenergic agonists to inhibit leptin expression and secretion. The inverse relationship between leptin secretion and the increase of glycerol levels, which is an index of the activation of cAMP-dependent protein kinases, suggests that activation of the cAMP signaling pathway mediates the inhibitory effects of Trecadrine on leptin gene expression and secretion. PMID- 12080444 TI - Relationship between peroxisome proliferator-activated receptor gamma and retinoic acid receptor alpha gene expression in obese human adipose tissue. AB - OBJECTIVE: To investigate in human adipose tissue a possible relationship between per oxisome proliferator-activated receptor gamma (PPARgamma) and retinoic acid receptor alpha (RARalpha) gene expression, two genes involved in the control of adipocyte differentiation. SUBJECTS: Ten lean control women (age 31-60 y, body mass index (BMI) 18-24.7 kg/m(2)) and an obese group of 15 women (age 27-62 y, BMI 30-57.5 kg/m(2)), of whom 10 subjects were in weight-gain phase and five were in weight-loss phase. MEASUREMENTS: We assessed the relative PPARgamma and RARalpha mRNA levels in subcutaneous abdominal adipose tissue using a real-time PCR method. RESULTS: PPARgamma mRNA level were significantly increased (+91%; P<0.01) in obese women compared to lean control women. In the obese group, we observed a PPARgamma mRNA level 42% lower in weight-loss obese than in weight gain obese subjects. We obtained a positive correlation (r=0.56; P<0.01) between PPARgamma mRNA level and the BMI of all subjects. Relative mRNA abundance level of RARalpha in subcutaneous adipose tissue of obese subjects is significantly lower than in control subjects (-56%, P<0.01), and a negative correlation was found between PPARgamma and RARalpha mRNA levels in subcutaneous adipose tissue of subjects study (r=-0.75; P<0.01). CONCLUSION: Our findings suggest that obesity is associated with an inverse relationship between PPARgamma and RARalpha expressions in human subcutaneous adipose tissue. Modulations of nuclear receptor profile could be an important event in the body's early adaptive mechanisms promoting adipose tissue plasticity and leading to the onset of obesity. PMID- 12080445 TI - Influence of the beta2-adrenergic receptor Arg16Gly polymorphism on longitudinal changes in obesity from childhood through young adulthood in a biracial cohort: the Bogalusa Heart Study. AB - BACKGROUND: The beta2-adrenergic receptor (ADRB2) plays a major role in regulating energy expenditure by stimulating lipid metabolism in human adipose tissue. Polymorphisms in the ADRB2 gene have been associated with obesity and various weight-related traits in cross-sectional studies of adults, but little is known about the effects of the ADRB2 gene on childhood obesity or the propensity to gain weight over time. OBJECTIVE: To assess the effects of a polymorphism in codon 16 (Arg16-->Gly) of the ADRB2 gene, which has been associated with a decrease in beta2-receptor density and efficiency, on longitudinal changes in obesity from childhood to young adulthood in a biracial cohort. DESIGN: Seven cross-sectional screenings of children and five cross-sectional screenings of young adults who were previously examined as children produced longitudinal data from childhood to young adulthood. METHODS: Height, weight and subscapular and triceps skinfolds were measured by trained examiners following identical protocols over the course of the study. Gender- and age-stratified analyses using random coefficients models were used to examine longitudinal genetic effects on obesity in 1151 African-American and Caucasian males and females who attended an average of six examinations over a 24 y period from childhood to young adulthood. RESULTS: Age-stratified analyses showed no clear genetic relationships with changes in obesity measures over time in females, but an age-dependent association was observed in males, where the relationship between the Arg16Gly polymorphism and obesity became stronger with age. In males who were 4-9 y of age at the beginning of the study in 1973, body mass index (BMI) was 4% higher in Gly/Gly and Arg/Gly males compared to those with Arg/Arg by 26 y of age. Subscapular skinfold measurements in Gly/Gly males became significantly different from Arg/Arg males (20% higher) by age 20. In the oldest male cohort (10-14 y of age in 1973), BMI increased at a significantly greater rate (0.4%/y) in males carrying the Gly16 form of the receptor relative to Arg/Arg males. BMI was significantly different between homozygous genotypes by approximately 26 y of age, and reached 8% higher in Gly/Gly males by age 32. Subscapular skinfolds also increased at a significantly greater rate (2%/y) in Gly/Gly males compared to Arg/Arg males, becoming significantly different (27%) by approximately 22 y of age and reaching a maximum difference of 50% by age 32. CONCLUSIONS: Our data suggest that the beta2-adrenergic receptor is associated with the propensity to gain weight from childhood to young adulthood in males. An increased understanding of genetic influences on the development of obesity may improve the effectiveness of interventions designed to reduce excess body weight and help define the role of genetic factors in diabetes and cardiovascular disease. PMID- 12080446 TI - Allelic variants in the GABA(A)alpha6 receptor subunit gene (GABRA6) is associated with abdominal obesity and cortisol secretion. AB - INTRODUCTION: Cortisol is involved in the regulation of adipose-tissue differentiation, function and distribution, and in excess causes abdominal obesity. At the level of the brain, cortisol secretion is partly controlled by gamma-aminobutyric acid (GABA), which is the major inhibitory neurotransmitter in the vertebrate brain, and acts by binding to GABA(A) receptors. METHOD: We examined the potential impact of a 1519T>C polymorphism in the GABA(A)alpha6 receptor subunit (GABRA6) gene on obesity and obesity-related phenotypes as well as circulating hormones, including salivary cortisol in 284 unrelated Swedish men born in 1944. The subjects were genotyped by using PCR amplification of the 3' non-coding region of the GABRA6 gene followed by digestion with the restriction enzyme AlwNI. RESULTS: The frequency of allele T was 0.54 and 0.46 for allele C. Carriers for the T allele (n=211) had borderline significantly higher waist-to hip ratio (P=0.094) and abdominal sagittal diameter (P=0.084) compared to homozygotes for the C allele (n=56). The homozygotes for the T allele had, in comparison to heterozygotes, significantly (P=0.004-0.024) higher mean cortisol levels at 11:45 am, and 30, 45 and 60 min after a standardized lunch and, finally, at 5:00 pm. In addition, T/T subjects had significantly (P=0.031) higher diurnal cortisol secretion compared to T/C subjects. Other hormones, glucose and serum lipids were not different across the genotype groups. CONCLUSION: These findings suggest a role of the 1519T>C polymorphism in GABRA6 in the predisposition to hypercortisolism and perhaps abdominal obesity. The pathophysiology may involve various environmental factors, particularly stress, that destabilize the GABA-hypothalamic-pituitary-adrenal systems in those with genetic vulnerability. PMID- 12080447 TI - Importance of plasma leptin in predicting future weight gain in obese children: a two-and-a-half-year longitudinal study. AB - OBJECTIVE: To determine whether relatively low leptin levels predict changes in adiposity in prepubertal and pubertal obese children. RESEARCH METHODS AND PROCEDURES: In a biracial cohort of 68 obese children (33 male and 35 female; 46 Caucasians and 22 African-Americans, age range 7-18 y), we measured at baseline fasting insulin and leptin levels, height and weight and calculated body mass index (kg/m(2)) and expressed body mass index as (BMI) Z-score. After a 2.5-y follow-up, anthropometric measurements were repeated and changes in weight gain were calculated as changes in BMI Z-score. RESULTS: At baseline obese preadolescent boys and girls had similar age and BMI Z-score, fasting insulin and leptin levels. After an average follow-up of 2.5 y, mean weight change calculated by changes in BMI Z-score from baseline was similar in both groups. In obese adolescent boys and girls at baseline, no significant gender differences were observed for BMI Z-score and insulin levels. In contrast, plasma leptin levels were significantly higher in obese girls compared with obese adolescent boys. At follow-up, there was no significant difference in change in BMI Z-score between obese boys and girls. Multiple linear regression analysis revealed that high basal leptin levels were positively associated with greater changes in BMI Z score only in girls (r(2)=0.18, P<0.02), after adjusting for basal BMI Z-score, Tanner stage, years of follow-up and basal insulin. High basal leptin levels in girls explained 18% of the weight gain. CONCLUSION: High leptin levels are associated with excessive future weight gain only in girls. PMID- 12080448 TI - Adjustment of fat-free mass and fat mass for height in children aged 8 y. AB - OBJECTIVE: To explore the relationships between height and (a) fat-free mass (FFM) and (b) fat mass (FM) in children in order to determine the optimum means of adjusting body composition for height. DESIGN: Cross-sectional study. SUBJECTS: Sixty-nine children aged 8 y. MEASUREMENTS AND METHODS: Weight and height (HT) were measured, and total body water by deuterium dilution for estimation of fat-free mass and fat mass. The indices FFM/HT(2) and FM/HT(2) were calculated, as were the indices FFM/HT(p) and FM/HT(p) where P was selected in order to eliminate the correlation of these indices with height. RESULTS: FFM was optimally adjusted for height by calculating FFM/HT(2). FM was optimally adjusted by calculating FM/HT(6). However, height accounted for <8% of the variation in FM/HT(2), indicating that the bias of this simpler index is small. CONCLUSIONS: Different adjustments of FFM and FM for height are possible, depending on the study design. The indices FFM/HT(2) and FM/HT(2) are appropriate for many purposes, and have the advantage of expressing both aspects of body composition in common units. However, in some scenarios a more sophisticated approach is required for evaluating body fatness. PMID- 12080449 TI - Fat-free mass index and fat mass index percentiles in Caucasians aged 18-98 y. AB - OBJECTIVE: To determine reference values for fat-free mass index (FFMI) and fat mass index (FMI) in a large Caucasian group of apparently healthy subjects, as a function of age and gender and to develop percentile distribution for these two parameters. DESIGN: Cross-sectional study in which bioelectrical impedance analysis (50 kHz) was measured (using tetrapolar electrodes and cross-validated formulae by dual-energy X-ray absorptiometry in order to calculate FFMI (fat-free mass/height squared) and FMI (fat mass/height squared). SUBJECTS: A total of 5635 apparently healthy adults from a mixed non-randomly selected Caucasian population in Switzerland (2986 men and 2649 women), varying in age from 24 to 98 y. RESULTS: The median FFMI (18-34 y) were 18.9 kg/m(2) in young males and 15.4 kg/m(2) in young females. No difference with age in males and a modest increase in females were observed. The median FMI was 4.0 kg/m(2) in males and 5.5 kg/m(2) in females. From young to elderly age categories, FMI progressively rose by an average of 55% in males and 62% in females, compared to an increase in body mass index (BMI) of 9 and 19% respectively. CONCLUSIONS: Reference intervals for FFMI and FMI could be of practical value for the clinical evaluation of a deficit in fat-free mass with or without excess fat mass (sarcopenic obesity) for a given age category, complementing the classical concept of body mass index (BMI) in a more qualitative manner. In contrast to BMI, similar reference ranges seems to be utilizable for FFMI with advancing age, in particular in men. PMID- 12080450 TI - The effects of body mass index on age at menopause. AB - OBJECTIVE: We examined which of body mass index (BMI, kg/m(2)), serum cholesterol (mg/dl), or systolic blood pressure (SBP, mm Hg) affected age at natural menopause. DESIGN: A population-based follow-up program. METHODS: We determined the age at natural menopause in 1136 women followed biennially since their first examination in 1958-1959 through the 16th examination in 1988-1989. Four-hundred and ninety-three naturally menopausal women were classified into three groups by BMI, serum cholesterol and SBP measurement levels at age 40 or 41 y: the upper 25%, middle 50%, and lower 25%. We then studied whether there was a difference in age at menopause among the three groups thus classified. The 1136 natural menopausal women were also classified as early (n=454; 45-49 y at menopause (48.3+/-1.2 y)) or late (n=682; >or=50 y at menopause (52.3+/-1.6 y)) menopausal and compared for premenopausal trends in BMI, serum cholesterol and SBP in the early and late menopausal women by means of a longitudinal data analysis model. RESULTS: When women were classified into the three groups based on a BMI that was measured at 40 or 41 y, age at menopause in the upper 25% (50.4+/-2.8 y) was significantly higher (P<0.05) than that in the lower 25% (49.7+/-2.8 y). The entire premenopausal trend in BMI in late menopausal women shifted upward compared to that in early menopausal women. On the other hand, the premenopausal trend more than 4 y before menopause in serum cholesterol and the entire premenopausal trend in SBP in late menopausal women were identical to those in early menopausal women. CONCLUSION: Among the variables studied, only BMI is related to age at menopause, and the greater the BMI, the later the age at menopause. PMID- 12080451 TI - Prevalence and correlates of large weight gains and losses. AB - OBJECTIVE: To examine the prevalence and correlates of large weight gains and losses over a 3 y period in a heterogeneous population of participants in a study of weight gain prevention. DESIGN AND MEASURES: Analyses based on a cohort of 823 participants in a weight gain prevention study whose weight was measured on at least two of four annual examinations. RESULTS: Weight gains and losses of >or=5% body weight over 1 y were observed in 9.3 and 15% of the population, respectively. Weight gains among those initially losing weight were significantly greater (3.9 kg) than among those experiencing stable weight (0.8 kg) or a large weight gain (1.5 kg) over the following 2 y. Cumulative weight changes over 3 y were -2.6, 1.0 and 7.6 kg among large loss, weight stable and large gain groups, respectively. Large weight loss was more common in smokers, large gains were more common in younger people and in those with a more extensive weight loss history, and stable weight was observed more often in individuals with less extensive histories of weight loss. CONCLUSION: The high prevalence of large short-term weight gains and losses in this heterogeneous population, their apparent resistance to short-term reversal, and the strength of their relationship to longer-term weight trends suggest that rapid weight change over relatively short time intervals is a phenomenon that deserves more research attention. Short periods of rapid weight gain may contribute importantly to rapidly rising obesity rates. PMID- 12080452 TI - Acquired preference especially for dietary fat and obesity: a study of weight discordant monozygotic twin pairs. AB - OBJECTIVE: To determine the independent associations of dietary preference for fat with obesity without the confounding by genetic effects. DESIGN: Descriptive comparison of the responses of monozygotic twins discordant for obesity to questions concerning current and past preference for dietary fat, current overconsumption of fatty items and recalled food consumption compared to the co twin. SETTING: The Research and Development Centre of the Social Insurance Institution, Finland. SUBJECTS: Twenty-three healthy monozygotic twin pairs who were discordant for obesity (BMI difference at least 3 kg/m(2)). MAIN OUTCOME MEASURES: Obesity status of the twin, as a function of the current and recalled dietary preferences and selected psychosocial variables. RESULTS: The obese twins reported current preference for fatty foods three times more frequently than the lean co-twin. Moreover, when comparing recalled taste for fat at the time the twins left their parental homes, both the obese and lean co-twins consistently recalled that the obese twin had greater preference for fatty foods in young adulthood, and that the lean twin had less. Psychological characteristics of lean and obese co-twins did not differ. CONCLUSION: Acquired preference for fatty foods is associated with obesity, independent of genetic background. Modification of fat preferences may be an important step in the prevention of obesity in the general population. PMID- 12080453 TI - Are dual-energy X-ray absorptiometry regional estimates associated with visceral adipose tissue mass? AB - OBJECTIVE: Recent studies support the possibility of estimating abdominal fat using a region of interest (ROI) selected by conventional whole body dual-energy X-ray absorptiometry (DXA). This is an important observation as DXA ROI estimates have some advantages over waist circumference or computed tomography/magnetic resonance imaging (MRI) as a means of assessing visceral adipose tissue (VAT) and adipose tissue distribution. The aim of this study was to evaluate the usefulness of DXA abdominal ROI estimates in assessing VAT among non-obese men. DESIGN: Observational, cross-sectional study comparing correlations between MRI-measured total VAT and surrogate measures including DXA ROIs. A stepwise multiple regression model was applied to derive a predictive equation with total VAT mass. SUBJECTS: Ninety non-obese healthy men between the ages of 18 and 44 y with BMI<30 kg/m(2). MEASUREMENTS: Abdominal adipose tissue and total VAT were measured by whole body MRI; VAT area by single-slice MRI at the L4-5 level; specific DXA ROIs for abdominal regional fat defined as ROI A (L2-4), B (L2-upper iliac), C (lower costal-upper iliac), and D (ROI C excluding spine); and simple anthropometric measures. RESULTS: Correlations between total VAT and ROIs A (r=0.85) and B (r=0.84) were not significantly different from that of VAT area at L4-5 (r=0.87), but significantly higher (P <0.01) than that of waist circumference (r=0.77). The highest correlations with total abdominal adipose tissue were for DXA ROIs and conventional DXA trunk fat (r=0.95-0.97). A stepwise multiple regression analysis revealed that 86% of the variance in total VAT was predicted by VAT area at L4-5, ROI A, and waist-hip ratio. CONCLUSIONS: DXA ROIs (L2-4, L2-upper iliac) were associated with total VAT as well as MRI-derived VAT area at L4-5 in non-obese men. DXA ROI fat distribution estimates may be useful in the early detection of men with abdominal/visceral obesity. PMID- 12080454 TI - The prediction of visceral fat by dual-energy X-ray absorptiometry in the elderly: a comparison with computed tomography and anthropometry. AB - INTRODUCTION: Effective methods for assessing visceral fat are important to investigate the role of visceral fat for the increased health risks in obesity. Techniques for direct measurement of soft tissue composition such as CT or MRI are expensive, time-consuming or require a relatively high radiation dose. Simple anthropometric methods, such as waist-to-hip ratio, waist circumference or sagittal diameter are widely used. However, these methods cannot differentiate between visceral and subcutaneous fat and are less accurate. The aim of the present study is to investigate whether the dual-energy X-ray absorptiometry (DXA) method, possibly combined with anthropometry, offers a good alternative to CT for the prediction of visceral fat in the elderly. METHODS: Subjects were participants in the Health ABC-study, a cohort study of black and white men and women aged 70-79, investigating the effect of weight-related health conditions on disablement. Total body fat and trunk fat were measured by DXA using a Hologic QDR 1500. A 10 mm CT scan at the L4-L5 level was acquired to measure visceral fat and total abdominal fat. Weight, height, sagittal diameter and waist circumference were measured using standard methods. Fat in a manually defined DXA subregion (4 cm slice at the top of iliac crest) at the abdomen was calculated in a sub-group of participants (n=150; 50% male; 45.3% Afro-American/54.7% Caucasian, age 70-79 y). This subregion, the standard trunk region and total fat were used as indicators of visceral fat. RESULTS: Total abdominal fat by DXA (subregion) was strongly correlated with total abdominal fat by CT (r ranging from 0.87 in white men to 0.98 in black women). The DXA subregion underestimated total abdominal fat by 10% compared to the CT slice. The underestimation by DXA was seen especially in people with less abdominal fat. The association of visceral fat by CT with the DXA subregion (r=0.66, 0.78, 0.79 and 0.65 for white and black men and women, respectively) was comparable with the association of the CT measure with the sagittal diameter (r=0.74, 0.70, 0.84 and 0.68). Combining DXA measurements with anthropometry gave only limited improvement for the prediction of visceral fat by CT compared to univariate models (maximal increase of r(2) 4%). CONCLUSION: DXA is a good alternative to CT for predicting total abdominal fat in an elderly population. For the prediction of visceral fat the sagittal diameter, which has a practical advantage compared to DXA, is just as effective. PMID- 12080455 TI - The central roles of obesity-associated dyslipidaemia, endothelial activation and cytokines in the Metabolic Syndrome--an analysis by structural equation modelling. AB - HYPOTHESIS: The multi-faceted components of the metabolic syndrome now include markers of inflammation and endothelial activation. Despite this growing body of epidemiological data, standard statistical methods fail to evaluate the nature of these associations adequately. In this pilot study, we hypothesize that obesity may lead to endothelial activation which is in part mediated by dyslipidaemia and proinflammatory cytokines. These factors interact to give rise to hyperinsulinaemia, hypertension and an anti-fibrinolytic state. To test this hypothesis, we used confirmatory factor analysis and structural equation modelling to fit these data to a model designed on theoretical grounds. METHODS: Metabolic syndrome variables, cytokines (IL6 and TNFalpha), markers of inflammation and endothelial activation were measured in 107 Caucasian non diabetic subjects aged 40-75 y. Using confirmatory factor analysis, we identified six factors to represent composite measurements of blood pressure, obesity, dyslipidaemia, hyperinsulinaemia, endothelial activation and the anti fibrinolytic state. We fitted these variables to two separate models, one using IL-6 and the other TNFalpha as the cytokine, and examined the inter-relationships (path analysis) amongst these variables, based on the above hypothesis. RESULTS: Men were centrally more obese and had increased markers of endothelial activation, inflammation and the anti-fibrinolytic state as well as hyperinsulinaemia and dyslipidaemia, compared with women. Obesity indexes (both body mass index and waist-hip ratio) were strongly associated with multiple cardiovascular risk factors. Both IL6 and TNFalpha were correlated with age, male gender, obesity indexes and markers of endothelial activation. Only IL-6 was associated with smoking while TNFalpha was correlated with hyperinsulinaemia. In the TNFalpha model, 61% of the obesity variance was explained by male gender, 36% of TNFalpha variance by age and dyslipidaemia, 43% of dyslipidaemia variance by age and obesity, 33% of hyperinsulinaemia variance by dyslipidaemia and a non smoking state, 29% of anti-fibrinolytic state variance by hyperinsulinaemia, 65% of endothelial activation variance by TNFalpha, dyslipidaemia and hyperinsulinaemia, 34% of blood pressure variance by hyperinsulinaemia and endothelial activation. In the IL-6 model, we observed similar relationships except that 23% of IL6 variance was explained by smoking and age. CONCLUSIONS: Using confirmatory factor analysis and structural equation modelling, we found that obesity, dyslipidemia and cytokines were the principal explanatory variables for the various components of the metabolic syndrome, with IL6 and TNFalpha having different explanatory variables and effects. These complex inter relationships were in part mediated by hyperinsulinaemia and endothelial activation. While this hypothetical model was based on scientific evidence, supported by rigorous analysis, it requires further confirmation in large-scale prospective studies. Given the complexity of the biological system and its interactions with exogenous factors, structural equation modelling provides a useful scientific tool for hypothesis testing, complementary to the more traditional experimental and cohort studies. PMID- 12080456 TI - Physical activity and its relationship with obesity, hypertension and diabetes in urban and rural Cameroon. AB - OBJECTIVES: To evaluate and compare physical activity patterns of urban and rural dwellers in Cameroon, and study their relationship with obesity, diabetes and hypertension. METHODS: We studied 2465 subjects aged >or=15 y, recruited on the basis of a random sampling of households, of whom 1183 were urban dwellers from Yaounde, the capital city of Cameroon and 1282 rural subjects from Bafut, a village of western Cameroon. They all had an interviewer-administered questionnaire for the assessment of their physical activity and anthropometric measurements, blood pressure and fasting blood glucose determination. The procedure was satisfactorily completed in 2325 (94.3%) subjects. Prevalences were age-adjusted and subjects compared according to their region, sex and age group. RESULTS: Obesity was diagnosed in 17.1 and 3.0% urban and rural women, respectively (P<0.001), and in 5.4 vs 1.2% urban and rural men, respectively (P<0.001). The prevalence of hypertension was significantly higher in urban vs rural dwellers (11.4 vs 6.6% and 17.6 vs 9.1% in women and men, respectively; P<0.001). Diabetes was more prevalent in urban compared to rural women (P<0.05), but not men. Urban subjects were characterized by lower physical activity (P<0.001), light occupation, high prevalence of multiple occupations, and reduced walking and cycling time compared to rural subjects. Univariate analysis showed significant associations between both physical inactivity and obesity and high blood pressure. The relationship of physical inactivity with hypertension and obesity were independent in both urban and rural men, but not in women. Body mass index, blood pressure and glycaemia were higher in the first compared with the fourth quartiles of energy expenditure. CONCLUSION: Obesity, diabetes and hypertension prevalence is higher in urban compared to rural dwellers in the populations studied. Physical activity is significantly lower and differs in pattern in urban subjects compared to rural. Physical inactivity is associated with these diseases, although not always significant in women. PMID- 12080457 TI - The influence of individual choices and attitudes on adiposity. AB - OBJECTIVE: To determine which dietary and lifestyle choices and which attitudes toward diet and health most contribute to adiposity and whether those factors differ by gender. METHODS: Multiple regression was used to estimate the effect of energy intake, energy expenditure, resources, demographics, and attitudes toward diet and health on self-reported body mass index (BMI) in a nationally representative sample of the US population. SAMPLE: This study used secondary data from the 1994-1996 Continuing Survey of Food Intakes by Individuals (CSFII) and its telephone follow-up, the Diet and Health Knowledge Survey (DHKS). CSFII is a nationally representative survey of individuals which obtained 24 h recalls of all food intake on two nonconsecutive days. The DHKS data set obtained information on knowledge and attitudes toward dietary guidance and health from individuals 20 y of age and over, who participated in the CSFII. Our sample consisted of responses from 5512 individuals. RESULTS: We found that many of the variables previously found to or hypothesized to influence body weight do so. Our analysis shows that choices and attitudes do matter for adiposity. Moreover, the analysis shows that the choices males and females make often have quite different effects on BMI. For females all the variables we constructed to represent energy intake, energy expenditure, and financial and human resources are significant. Most of the variables that characterize attitudes toward diet and health are significant. For males, far fewer variables are significant. Our tests for gender related differences show that whether or not males believe their weight is predetermined has no influence on BMI. In contrast, women who do not believe that their weight is predetermined have lower BMI values than those who believe weight is genetically determined. DISCUSSION: While our qualitative conclusion--choices and attitudes affect BMI--is unsurprising, the quantitative aspects are important. The analysis shows that factors affecting BMI differ systematically by gender. Understanding these systematic differences is critical to the design of information campaigns. Our analysis suggests that entirely different campaigns are necessary to communicate the importance of diet and lifestyle choices to men and to women. PMID- 12080458 TI - Hysteria. PMID- 12080459 TI - Neurogenic heterotopic ossification in spinal cord injury. AB - Neurogenic heterotopic ossification (NHO) is a frequent complication in spinal cord injury (SCI) that is often difficult to treat. This review emphasizes the incidence, risk factors and clinical signs of NHO in SCI patients. Although the exact pathophysiology underlying NHO in neurologic patients is not yet understood, different pathogenic mechanisms have been proposed in the literature. A selection of the most important theories will be given and discussed. Moreover the different diagnostic, therapeutic, and preventive methods currently used in NHO management after SCI will be reviewed. PMID- 12080460 TI - Conversion motor paralysis disorder: overview and rehabilitation model. AB - It is important to consider a differential diagnosis between paralysis on an organic basis and paralysis and disability due to psychological mechanisms in people with physical impairment secondary to trauma, without evidence of organic etiology. We review the most dramatic type of conversion disorder (CD) 'Conversion Motor Paralysis'. Recent important medical literature concerning the accepted treatment and rehabilitation management will be reviewed and discussed. The inter-disciplinary in-patient team management approach in a rehabilitation setting offers the benefits of a comprehensive assessment and treatment. The diagnosis is temporary and conditional, since there may be a long delay until the appearance of organic findings. A complete medical assessment is essential in order to rule out any possibility of an organic etiology. In as many as 25% to 50% of patients diagnosed as conversion, an organic medical diagnosis was found. PMID- 12080461 TI - Conversion motor paralysis disorder: analysis of 34 consecutive referrals. AB - STUDY DESIGN: We present our cumulative experience with patients sustaining the most dramatic type of Conversion Disorder (CD) - Conversion Motor Paralysis. SETTING: Rehabilitation departments, Reuth Medical Center, Tel-Aviv and Sheba Medical Center, Tel-Hashomer, Israel. METHODS: During the period 1973-2000, 34 patients with neurological symptoms without any anatomical or physiological basis were admitted to both rehabilitation departments. This number consists of less than 1% of the total acute traumatic and non-traumatic spinal cord disorders admitted annually to these centers. RESULTS: Twenty-five of the subjects were men (mean age of 30 years) and nine were women (mean age of 31.4 years). Neurological symptoms included: paraplegia (complete or incomplete) (18), hemiplegia or hemi paresis (11), tetraplegia (complete or incomplete) (three), monoplegia (one), triplegia (one). The final diagnosis on discharge was CD in 30 of the 34 cases, the remaining four being diagnosed as malingering. Functionally, nine patients had a complete recovery, 10 a partial recovery and 15 remained unchanged. CONCLUSION: Disabled people who experienced traumatic events resulting in various disabilities are admitted usually to a rehabilitation center. However, some of them are later diagnosed as having Conversion Disorder or malingering. We believe that their participation in active regular and integrative rehabilitation process is beneficial to most of them. Most of these patients gain functional independence and return to the main stream of life. PMID- 12080462 TI - Measuring quality of life of persons with spinal cord injury: external and structural validity. AB - STUDY DESIGN: Measurement evaluation of the external and structural components of validity. OBJECTIVES: To examine the relationships between quality of life (QOL) as measured by the spinal cord injury (SCI) version of the Ferrans and Powers Quality of Life Index (QLI) and other constructs represented within the model of disablement; and to examine the domains and scoring model of the QLI by exploring item and overall score/section score relationships. SETTING: Community, Alberta, Canada. METHODS: A convenience sample of 98 individuals with SCI living in the community completed the QLI and measures representing the model of disablement including the ASIA motor index, Functional Independence Measure, Reintegration to Normal Living index, Rosenberg's Self-Esteem Scale and Rotter's Internal-External Locus of Control scale. RESULTS: Four of the five a priori hypotheses were supported. Locus of control was not significantly related to QOL as expected. Factor analysis resulted in a five-factor structure that differed from the four domain model of the original QLI. Scoring relationships indicated that both the satisfaction and importance ratings contribute to the overall score, although not equally. CONCLUSION: There is support for the external component of validity although further examination regarding locus of control for persons with SCI is warranted. The structural component of validity requires further investigation to elucidate the domains of the SCI version of the QLI and the contribution of the importance scores. PMID- 12080463 TI - A tale of two countries: environmental impacts on social participation after spinal cord injury. AB - STUDY DESIGN: Comparative analysis of survey data produced in two countries. OBJECTIVES: To assess the degree to which environmental barriers impact social participation, and to identify the aspects of participation most affected. SETTING: Community-dwelling individuals with spinal cord injury in the USA and Turkey. METHODS: Subjects completed the Functional Independence Measure (FIM) motor score, the Craig Hospital Inventory of Environmental Factors (CHIEF), and the Craig Handicap Assessment and Reporting Technique (CHART). Analysis of co variance was used to analyze CHIEF and CHART differences within and between country. RESULTS: US subjects reported higher participation scores (CHART) and lower barriers (CHIEF), however, when controls for age, gender, time since injury and motor ability (FIM) were applied, country differences in reports of barriers were limited. Motor ability was the major predictor of participation, which was minimally affected by barriers. CONCLUSION: Conceptualization and measurement issues may have been the reason for the minimal support for the hypothesis that environment affects participation. Suggestions for future research are made. SPONSORSHIP: National Institute on Disability and Rehabilitation Research, Centers for Disease Control and Prevention. PMID- 12080464 TI - Neurological outcome following laminectomy in spinal metastases. AB - STUDY DESIGN: Retrospective outcome measurement study. OBJECTIVES: Patients suffering from malignant tumour disease and metastases to the spine develop a variety of clinical complaints including radicular symptoms and/or spinal cord compression syndromes. Palliative decompressive laminectomy with total or partial tumour resection is an acknowledged method of treatment, despite controversy. SETTING: The Department of Neurosurgery of the University of Vienna. METHOD: Patients suffering from metastases with predominant infiltration of the dorsal epidural parts, or patients who could not be operated on via an anterior approach, were included. Eighty-four patients who met these criteria underwent decompressive laminectomy with total or partial tumour removal. The study analyzed motor function, pain relief and continence in a 2- and 4-month post operative follow-up. According to the criteria of motor performance, 20% of the patients had been mobile pre-operatively. RESULTS: In the immediate post operative period 45%, after 2 months 33% and after 4 months 26%, were considered mobile. None of the paraplegic patients showed functional improvement. The median survival time was 6.5 months. Pre-operatively, 56% of the patients had shown continence dysfunction. Post-operatively, 38%, and after 2 months 46% of the patients, developed continence disorders. A significant reduction in analgesic medication was also observed in the post-operative period. CONCLUSION: In our series, palliative laminectomy with total or subtotal tumour reduction in patients with malignant spinal metastatic disease resulted in amelioration of motor function, pain and continence and therefore improved the patients' quality of life. The improvement in quality of life shows that this method is a valuable option in neurosurgical therapy, except for cases with pre-operative paraplegia. However, in patients with severe pre-operative paraparesis, the authors recommend laminectomy only in very exceptional cases, because of the poor post-operative neurological results. PMID- 12080465 TI - Paraplegia: an unusual presentation of Ewing's sarcoma. AB - STUDY DESIGN: Single-subject case. OBJECTIVES: To describe the atypical presentation of a rare tumor and difficulties in diagnosis. SETTING: Spinal cord unit of a rehabilitation and care center in Ankara, Turkey. METHODS: A 22-year old male patient with paraplegia was admitted to our center for rehabilitation. He underwent various diagnostic procedures to explain his clinical situation. RESULTS: Abdominal ultrasound and computed tomography revealed a mass and vertebral destruction. The pathology report confirmed the diagnosis of Ewing's sarcoma. CONCLUSION: Spinal cord compression is an important complication of primary or metastatic malignant tumors. Although it is a late complication in most cases, some tumors including Ewing's sarcoma may present with paraplegia. Trauma may be found in the patient's history. An atypical clinical course in a musculoskeletal or neurological condition should alert us to a possible underlying malignant disease. PMID- 12080466 TI - The tumor suppressor activity of SOCS-1. AB - SOCS-1 is an inducible SH2-containing inhibitor of Jak kinases and as such can potently suppress cytokine signaling. SOCS-1 deficient mice die within the first three weeks of life from a myeloproliferative disorder driven by excessive interferon signaling. We report here that SOCS-1 inhibits proliferation signals induced by a variety of oncogenes active within the hematopoietic system. Ectopic expression of SOCS-1 abolished proliferation mediated by a constitutively active form of the KIT receptor, TEL-JAK2, and v-ABL, and reduced metastasis from BCR ABL transformed cells. SOCS-1, however, did not interfere with v-SRC or RASV12 mediated cellular transformation. A mutant form of SOCS-1 unable to bind through its SH2 domain to tyrosine phosphorylated proteins could still inhibit KIT, but not TEL-JAK2, indicating multiple mechanisms for SOCS-1-mediated tumor suppression. We show that the steady state levels of TEL-JAK2 and to a greater extent v-ABL are diminished in the presence of SOCS-1. Lastly, we show that SOCS 1 -/- fibroblasts are more sensitive than wild type fibroblasts to either spontaneous or oncogene-induced transformation. These data suggest that loss-of function of SOCS-1 may collaborate with a variety of hematopoietic oncogenes to facilitate tumor progression. PMID- 12080467 TI - Ubiquitination capabilities in response to neocarzinostatin and H(2)O(2) stress in cell lines from patients with ataxia-telangiectasia. AB - The human genetic disorder ataxia-telangiectasia (A-T) is due to lack of functional ATM, a protein kinase which is involved in cellular responses to DNA double strand breaks (DSBs) and possibly other oxidative stresses, as well as in regulation of several fundamental cellular functions. Studies regarding responses in A-T cells to the induction of DSBs utilize ionizing radiation or radiomimetic chemicals, such as neocarzinostatin (NCS), which induce DNA DSBs. This critical DNA lesion activates many defense systems, such as the cell cycle checkpoints. The cell cycle is also regulated through a timed and coordinated degradation of regulatory proteins via the ubiquitin pathway. Our recent studies indicate that the ubiquitin pathway is influenced by the cellular redox status and that it is the major cellular pathway for removal of oxidized proteins. Accordingly, we hypothesized that the absence of a functional ATM protein might involve perturbations to the ubiquitin pathway as well. We show here that upon treatment with NCS, there was a transient 50-70% increase in endogenous ubiquitin conjugates in A-T and wt lymphoblastoid cells. Ubiquitin conjugation capabilities per se and levels of substrates for conjugation were also similarly enhanced in wt and A-T cells upon NCS treatment. We also compared the ubiquitination response in A-T and wt cells using H(2)O(2) as the stress, in view of preexisting evidence of the effects of H(2)O(2) on ubiquitination capabilities in other types of cells. As with NCS treatment, there was an approximately 45% increase in endogenous ubiquitin conjugates by 2-4 h after exposure to H(2)O(2). Both cell types showed a rapid 50-150% increase in de novo formed 125I-ubiquitin conjugates. As compared with wt cells, unexposed A-T cells had higher endogenous levels of conjugates and enhanced conjugation capability. However, A-T cells mounted a more muted ubiquitination response to the stress. The enhanced ubiquitin conjugation in unstressed A-T cells and attenuated ability of these cells to respond to stress are consistent with the A-T cells being under oxidative stress and with their having an 'aged' phenotype. The indication that ubiquitin conjugate levels and ubiquitin conjugation capabilities are enhanced upon oxidative stress without significant changes in GSSG/GSH ratios indicates that assays of ubiquitination provide a sensitive measure of cellular stress. The data also add support to the impression that potentiated ubiquitination response to mild oxidative stress is a generalizable phenomenon. PMID- 12080468 TI - Transformation of Ba/F3 cells and Rat-1 cells by ETV6/ARG. AB - ETV6/ARG, a novel fusion gene composed of the ETV6 HLH oligomerization domain and most of sequences of the ARG protein tyrosine, was recently identified in human leukemia cells. The presence of the ETV6/ARG translocation raises the possibility that the resulting fusion protein functions as an oncogene. However, the transforming activity of the ETV6/ARG protein has not been determined and its contribution to leukemogenesis is therefore unknown. Here we address this question by analysing the oncogenic activity of ETV6/ARG in hematopoietic and fibroblast cells. It is demonstrated that expression of ETV6/ARG confers IL3 independent growth to Ba/F3 cells and anchorage independent growth to Rat-1 fibroblasts. It is also shown that multiple signaling molecules, including PI3K, SHC, ras-GAP and CRK-L, are tyrosine phosphorylated in Ba/F3 cells that express ETV6/ARG. Analysis of four different types of ETV6/ARG transcripts previously identified in the AML-M3 leukemia cell line HT93A suggest that ETV6 HLH domain is required for oncogenic activity. Based upon these results it is concluded that ARG can be activated as an oncogene in human malignancy and that the ETV6/ARG oncoprotein triggers some of the same signaling pathways associated with activated ABL oncogenes. PMID- 12080469 TI - Regulation of c-myc stability by selective stress conditions and by MEKK1 requires aa 127-189 of c-myc. AB - C-myc availability is central for its ability to serve as a regulator of cell growth and death. Here we study the regulation of c-myc protein stability and identify domains of c-myc that are important for its stabilization in response to stress kinases activated following selective stress conditions. UV-irradiation elicited an increase in c-myc protein levels, which could be attenuated by inhibitors of stress kinases but also by actinomycin D-inhibitor of transcription. Inhibition of protein synthesis results in a noticeable decrease in c-myc levels, further pointing to the short half-life of the protein. However, in combination with tumor necrosis factor-alpha (TNF-alpha), cycloheximide efficiently increases steady-state levels of c-myc, suggesting that selective stress conditions are required to increase c-myc protein stability. Expression of MEKK1, an upstream regulator of protein kinases that has been implicated in mediating the response to diverse stress conditions, also results in an efficient increase in the half-life of c-myc protein. To map c-myc domains that are responsive to stress kinases, we monitored changes in the level of c-myc deletion mutants following MEKK1 expression. Of the seven c-myc deletion mutants analysed, the domain spanning amino acids 127-189 was found to be required for MEKK1 dependent increase in c-myc stability. In all, the present study identifies a novel domain that is important for the regulation of c-myc stability by stress kinases in response to selective stress conditions. PMID- 12080470 TI - The c-Rel transcription factor can both induce and inhibit apoptosis in the same cells via the upregulation of MnSOD. AB - Rel/NF-kappaB transcription factors are involved in several physiological processes, including the regulation of apoptosis. These factors were shown to exhibit pro- or anti-apoptotic activities in different cellular models, but at present, the mechanisms underlying these opposite effects are poorly understood. In this study, we show that the constitutive expression of a transcriptionally active member of the Rel/NF-kappaB family, c-Rel, first induces a resistance against TNFalpha-induced apoptosis and later increases the level of spontaneous apoptosis of HeLa cells. Both the anti- and pro-apoptotic effects increase with the level of c-Rel overexpression. The up-regulation by c-Rel of the manganese superoxide dismutase (MnSOD) could explain both the rapid anti-apoptotic effect and the delayed pro-apoptotic one. Indeed, the enzymatic activity of MnSOD is to transform the toxic O(2)(*)(-) in H(2)O(2). Hence, on one hand, its induction helps cells to resist against the apoptogenic burst of O(2)(*)(-) produced upon TNFalpha stimulation, but on the other hand, it leads to a progressive H(2)O(2) accumulation that ultimately results in apoptosis. These results indicate that the anti- and pro-apoptotic effects of Rel/NF-kappaB factors are not necessarily alternative but can occur successively in the same cell, via the up-regulation of the same target gene. PMID- 12080471 TI - Identification of novel candidates for replicative senescence by functional proteomics. AB - To identify the underlying mechanisms that limit the mitotic potential of normal somatic cells, we have undertaken a high resolution differential proteomic analysis aimed at identifying proteins that were differentially expressed upon replicative senescence. Since replicative senescence in heterogeneous primary fibroblast cultures is asynchronous, we analysed a group of conditionally immortalized rat embryo fibroblast cell lines that have previously been shown to undergo synchronous senescence upon inactivation of SV40 tsA58 T antigen. This identified 43 spots that were differentially expressed in these cell lines. Comparison of the identity of these features with those identified in a complimentary independent differential proteomic analysis of replicative senescence, directly in primary rat embryo fibroblasts upon serial passaging, identified nine features that were in common between the two studies even though they had been conducted entirely separately. None of these proteins have previously been recognized to be involved with replicative senescence. Thus, they represent novel starting points for elucidating the underlying mechanism that regulates the finite mitotic life span of somatic cells and how it can be overcome in cancer cells. PMID- 12080472 TI - Mdm2 inhibition of p53 induces E2F1 transactivation via p21. AB - The transcription factor E2F1 functions as a key regulator for both cell-cycle progression and apoptosis. Mdm2, a major cellular regulator of the p53 tumor suppressor protein, is also closely involved in cell cycle and apoptosis. In addition to regulation of p53, Mdm2 has been reported to stimulate E2F1 transactivation by a mechanism that remains unclear. Here we examined how overexpression of Mdm2 alters E2F1/DP1 transactivation. Using a set of cell lines with differing p53 and Rb status we determined that Mdm2 induction of E2F1 transactivation was p53-dependent, resulting from release of repression by p53. While Mdm2 association with p53 was required to increase E2F1 transactivation, Mdm2 mediated degradation of p53 was not. p53 repression of E2F1 transactivation required a functional DNA binding and transactivation domain. Consistent with Mdm2 activation of E2F1 via an inhibition of p53 transactivation we demonstrate a concomitant reduction in p21 protein levels with Mdm2 overexpression. Furthermore, E2F1 repression by an Rb-phosphorylation mutant could not be reversed by Mdm2 overexpression. Mdm2 was also unable to enhance E2F1 transactivation in Mouse embryo fibroblasts lacking p21. Taken together, these results suggest that Mdm2 activation of E2F1 occurs through the repression of p53 dependent transcription of p21, a p53-target gene and cyclin dependent kinase inhibitor. PMID- 12080473 TI - Metastatic Lymph Node 51, a novel nucleo-cytoplasmic protein overexpressed in breast cancer. AB - Metastatic Lymph Node 51 (MLN51) cDNA was isolated by differential screening of a human breast cancer metastasis cDNA library. MLN51 cDNA encodes a novel human protein of 703 residues that shares no significant homology to any known protein. However MLN51 is well conserved between vertebrate and invertebrate species suggesting an important biological function. The amino terminal half of the protein contains a coiled-coil domain and two potential nuclear localization signals (NLS). The carboxy terminal half contains one SH2 and four SH3 binding motifs. The coiled-coil domain promotes MLN51 oligomerization in transfected cells. When transiently expressed, the MLN51 protein is mainly found in the cytoplasm with a weak nuclear staining. However, deletion of the carboxy terminal half of the protein allows the targeting of the protein to the nucleus, demonstrating that the NLSs are functional. MLN51 is ubiquitously expressed in normal tissues. Human breast carcinomas show MLN51 overexpression in malignant epithelial cells. The uncommon association of protein-protein interaction domains often found either in nuclear or in cytoplasmic signaling proteins raises a possible nucleo-cytoplasmic function for MLN51. PMID- 12080475 TI - R-Ras C-terminal sequences are sufficient to confer R-Ras specificity to H-Ras. AB - Activated versions of the similar GTPases, H-Ras and R-Ras, have differing effects on biological phenotypes: Activated H-Ras strongly transforms many fibroblast cell lines causing dramatic changes in cell shape and cytoskeletal organization. In contrast, R-Ras transforms fewer cell lines and the transformed cells display only some of the morphological changes associated with H-Ras transformation. H-Ras cells can survive in the absence of serum whereas R-Ras cells seem to die by an apoptotic-like mechanism in response to removal of serum. H-Ras can suppress integrin activation and R-Ras specifically antagonizes this effect. To map sequences responsible for these differences we have generated and investigated a panel of H-Ras and R-Ras chimeras. We found that the C-terminal 53 amino acids of R-Ras were necessary and sufficient to specify the contrasting biological properties of R-Ras with respect to focus morphology, reactive oxygen species (ROS) production and reversal of H-Ras-induced integrin suppression. Surprisingly, we found chimeras in which the focus formation and integrin mediated phenotypes were separated, suggesting that different effectors could be involved in mediating these responses. An integrin profile of H-Ras and R-Ras cell pools showed no significant differences; both activated H-Ras and R-Ras expressing cells were found to have reduced beta(1) activity, suggesting that the activity state of the beta(1) subunit is not sufficient to direct an H-Ras transformed cell morphology. PMID- 12080474 TI - CL100 expression is down-regulated in advanced epithelial ovarian cancer and its re-expression decreases its malignant potential. AB - Although early stage ovarian cancer can be effectively treated with surgery and chemotherapy, the majority of cases present with advanced disease, which remains essentially incurable. Unfortunately, little is known about the genes important for the development and progression of this disease. In this study, the expression of 68 phosphatases was determined in immortalized ovarian epithelial cells (IOSE) and compared to ovarian cancer cell lines. CL100, a dual specificity phosphatase, displayed 10-25-fold higher expression in normal compared to malignant ovarian cell lines. Immunohistochemical staining of normal ovaries and 68 ovarian cancer specimens confirmed this differential expression. Re-expression of CL100 in ovarian cancer cells decreased adherent and non-adherent cell growth and induced phenotypic changes including loss of filopodia and lamellipodia with an associated decrease in cell motility. Induced expression of CL100 in ovarian cancer cells suppressed intraperitoneal tumor growth in nude mice. These results show for the first time that CL100 expression is altered in human ovarian cancer, that CL100 expression changes cell morphology and motility, and that it suppresses intraperitoneal growth of human ovarian epithelial cancer. These data suggest that down-regulation of CL100 may play a role in the progression of human ovarian cancer. PMID- 12080478 TI - Interactions of stress and reproduction - a personal view. AB - In the first part of this article, the authors discuss the effect of stress upon reproduction. The paper begins with a discussion of the various stressful factors that influence the functions of reproductive organs under various circumstances, including menarche, gestation and lactation. In general, physical and emotional stress has a negative impact upon reproductive function. Chronic stress may delay pubescence and, during the reproductive years, may impair oogenesis and spermatogenesis. This process may result in transitory infertility in women and permanent sterility in men. If associated with pregnancy, stress may be conducive to spontaneous abortion or premature birth. The second part of the article deals with stressful stimuli attributable to the reproductive process, the problem of infertility, techniques of assisted reproduction, and the management of unwanted gestations including induced abortion, fetal reduction, spontaneous miscarriage as well as in utero fetal death. The stress generating effects of singleton and multiple pregnancies and births are also outlined in some detail. In the third part of this article discuss effect of stress upon the fetus directly or through the maternal organism. PMID- 12080476 TI - Allelic loss and reduced expression of the ING3, a candidate tumor suppressor gene at 7q31, in human head and neck cancers. AB - Loss of heterozygosity (LOH) has been frequently detected at chromosome 7q31 region in human head and neck squamous cell carcinomas (HNSCC) and many other cancers, suggesting the existence of tumor suppressor genes (TSG). We analysed LOH at 7q31 region in 49 HNSCC by using six polymorphic microsatellite markers and found allelic deletion in 48% (22/46) of the informative cases. We detected two preferentially deleted regions, one is around D7S643 and the other around D7S486. When we redefined the map of 7q31 region according to the contiguous sequences, a recently identified gene, ING3, was found in the proximity of D7S643. ING3 protein harbors the PHD domain highly homologous among ING family proteins, in which we previously found mutations in a related gene, ING1. As only one missense mutation of the ING3 gene was found in HNSCC, we examined the expression level. Reverse-transcription-PCR analysis demonstrated decreased or no expression of ING3 mRNA in 50% of primary tumors as compared with that of matched normal samples. Especially, about 63% of tongue and larynx tumors showed the decrease and a tendency of higher mortality was observed in cases with decreased ING3 expression. All these findings suggest a possibility that the ING3 gene functions as a TSG in a subset of HNSCC. PMID- 12080479 TI - [Is primary, secondary or tertiary prevention of coronary heart disease possible by means of oestrogens or gestagens?]. AB - Clinical trials in primary prevention of coronary atherosclerosis have demonstrated that estrogen application in postmenopausal women may reduce the relative risk by approximately 50 % when initiated at the time of decreasing endogenous estradiol production. Multifactorial estrogen actions (i. e. vasodilatation, inhibition of vascular smooth muscle cell proliferation) could explain these observations. Based on some investigations in secondary and tertiary prevention of coronary atherosclerosis in postmenopausal women it may be concluded that oestrogens alone or in combination with certain progestins may have a protective effect in reducing the rate of reinfarctions. From an epidemiological point of view it was emphasized that these studies were not randomised and therefore not suitable for drawing conclusions generally valid in all postmenopausal women. The HERS-study was initiated in order to get reliable informations. However, the results of this study are not comparable with any other investigation as in the HERS-study for the first time a continuous combined oestrogen/progestin application was chosen. The authors pointed out that several positive oestrogen actions may be antagonised by progestins as the oestrogen receptor density could be down regulated by progestins. The HERS study shows indeed no increase in reinfarctions and a decrease in total mortality after long term treatment. Consequently, the study shows that there is a demand to strongly emphasise the significance of primary prevention. PMID- 12080480 TI - [Differing aspects of human dignity in intensive medicine]. AB - Human dignity often is used as an ethical category in medical judgement of current medical and intensive care decisions. The short history of the intensive care medicine, however, demonstrates a certain drift in the use of the term "human dignity". Parallel to the more pronounced patient autonomy, articulated in patient testaments, therapeutic decisions of current intensive care medicine individualize therapeutic approaches, instead of preferentially maximizing life span. PMID- 12080481 TI - [Comparability of obstetric and perinatal data from England and Germany using standard-primip-groups - maternal and neonatal outcome]. AB - OBJECTIVE: Aim of the study was to assess whether different birth management of an english and german department can influence the maternal and neonatal outcome. MATERIAL AND METHODS: The database consisted of routinely published data from 1986-95 for two clinical units in Solihull (England) and Ibbenbueren (Germany) comprising 34 820 and 9 053 deliveries respectively. In order to standardise the obstetric risk profiles the heterogeneous primary groups were subdivided into "standard primip groups". A statistical comparison using the "binomial confidence interval method" was carried out. RESULTS: In the standardised comparison induction of labour, duration of labour 1-6 hours, vaginal delivery from cephalic presentation, elective caesarean section, both for cephalic and breech presentation, transfer to the childrens hospital were less frequent in Solihull. On the other hand, Solihull showed more frequent oxytocin administration, fetal blood analysis, epidural anaesthesia, episiotomies, duration of labour > 13 hours, forceps, ventouse and emergency caesarean section deliveries from cephalic presentation, vaginal deliveries or emergency caesarean sections from breech presentation, resuscitation of the newborn using mask and/or drugs, maternal blood loss > 1 000 ml as well as abnormalities of placental separation. Despite the different management of the departments being compared no significant differences in the incidence of perinatal hypoxia as determined by Apgar scores at 5 minutes nor in the fetal mortality rate between the units could be identified. CONCLUSION: Using standardised data the quality of obstetric and perinatal care in England and Germany can be reliably compared. Different birth management does not significantly influence the neonatal outcome. PMID- 12080482 TI - [Primary and secondary breast cancer prevention. Knowledge, assessment and participation among the female population of Schleswig-Holstein]. AB - OBJECTIVE: The intention was to find out to what extend women in Schleswig Holstein were informed about primary and secondary preventive measures concerning breast cancer, how they assess their efficiency and in how far they are willing to take part in them. From June 1999 to April 2000 1 520 women in Schleswig Holsteins were asked to complete a questionnaire about breast cancer prevention. There was a representative opinion poll of the market research company "Institut fur Gesundheitsforschung Munchen (Infratest)". RESULTS: Most of the women (90.6 %) believed in the efficiency of early recognition of breast cancer at the gynaecologist, only 79.3 % of them stated that they took the chance of taking part in the annual examination. According to the doctors Union "Kassenarztliche Vereinigung", only 50.4 % of the female members of health insurance schemes took part in examinations concerning the early recognition of breast cancer in 1999. The degree of knowledge about and participation in preventive measures depend on the age of the women. The gynaecologist was named as the most important source of information and counselling. CONCLUSION: In future the population needs more information and instruction concerning breast cancer preventive measures. PMID- 12080483 TI - A randomized controlled trial of levonorgestrel releasing IUD and thermal balloon ablation in the treatment of menorrhagia. AB - Our aim was to compare the treatment of menorrhagia either with a levonorgestrel releasing intrauterine device or with endometrial thermal balloon ablation. The primary endpoints of evaluation were menstrual blood flow reduction and the increase in hemoglobin values, while the secondary end points were adverse side effects; health related quality of life. After randomization, 36 women underwent outpatient thermal balloon ablation under local anesthesia and an intrauterine device releasing 20 microgram/day of levonorgestrel, were inserted within the first 7 days of menses to 36 women. Both techniques were found to be significantly effective in reducing the menstrual blood loss but in comparison thermal balloon ablation was more effective in Deltamean +/- SD decrease of pictorial sores (388.2 +/- 21 vs 343 +/- 27; p < 0.001). We noted a significant but similar increases in Deltamean +/- SD hemoglobin values (3.9 +/- 1.7 vs 3.7 +/- 1.4; p:.21). Patients treated by thermal balloon ablation reported fewer side effects and perceived a higher health related quality of life in physical role functioning. At one year of follow-up, the medicated device was effective but not as effective as thermal balloon ablation in reducing the menstrual blood loss. However it was found to be as effective as thermal balloon ablation in increasing the hemoglobin values. The side effect profile of the medicated device may alter its acceptability by reducing the perceived health related quality of life in menorrhagic women with no desire for further childbearing. PMID- 12080484 TI - Rupture of coliform pyosalpinx in a nine-year old girl. AB - The authors report on an atypical clinical picture of an acute abdomen syndrome caused by the rupture of an atypically located pyosalpinx in a 9-year old girl.A perforated right-sided pyosalpinx was found at two locations, firmly adhering to a vesicouterine excavation. Right-sided salpingectomy, appendectomy, pelvic cavity lavage and drainage of the Douglas cavity were performed. The pathohistology indicated a perforated pyosalpinx and catarrhal appendicitis. On bacteriological analysis, Escherichia coli was isolated from the vaginal discharge and from the pus collected from the small pelvis. PMID- 12080486 TI - [Analysis of the patients who missed their appointments at the neurology clinic of the Marina Alta]. AB - OBJECTIVES: To analyze the frequency, reasons and characteristics of patients who missed their appointments (NP) at the Neurology Clinic (CEN). To compare the characteristics of the NP with the patients who did not miss their appointment. Suggest measures to reduce avoidable causes of NP. PATIENTS AND METHODS: During the period 1/10/99 to 30/4/2000 the cases of NP were prospectively recorded and compared with a sample of patients who attended the CEN during the same period. We analyze the data on age, sex, distance of home from CEN, diagnosis, type of consultation and date of last consultation. The NP were contacted by phone and asked why they had not gone for their appointments. The reasons given were then classified as avoidable or unavoidable. RESULTS: Of the 1,842 consultations scheduled, 19% were NP. When the NP and the patients who did attend were compared we found statistically significant differences regarding the distance home/CEN. According to the diagnosis, the NP had more 'symptoms/signs' and other MIA, whilst those who attended the clinic had more 'neuropathies' and 'awaiting diagnosis'. The commonest avoidable reasons (55.8%) for NP were forgetting, administrative error and communication failure, and for unavoidable reasons (44.2%) physical disability, other priorities and improvement. CONCLUSIONS: There is a large proportion of NP in the CEN of the Marina Alta. The distance from home to CEN (over 25 km) was the main factor affecting the rate of NP. Since 56% of the NP were due to avoidable causes, strategies could be designed to reduce this percentage. In the NP group, the main causes were forgetting, administrative errors and communication failure. PMID- 12080485 TI - Mutations in two genes encoding different subunits of a receptor signaling complex result in an identical disease phenotype. AB - Polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy (PLOSL), also known as "Nasu-Hakola disease," is a globally distributed recessively inherited disease leading to death during the 5th decade of life and is characterized by early-onset progressive dementia and bone cysts. Elsewhere, we have identified PLOSL mutations in TYROBP (DAP12), which codes for a membrane receptor component in natural-killer and myeloid cells, and also have identified genetic heterogeneity in PLOSL, with some patients carrying no mutations in TYROBP. Here we complete the molecular pathology of PLOSL by identifying TREM2 as the second PLOSL gene. TREM2 forms a receptor signaling complex with TYROBP and triggers activation of the immune responses in macrophages and dendritic cells. Patients with PLOSL have no defects in cell-mediated immunity, suggesting a remarkable capacity of the human immune system to compensate for the inactive TYROBP-mediated activation pathway. Our data imply that the TYROBP-mediated signaling pathway plays a significant role in human brain and bone tissue and provide an interesting example of how mutations in two different subunits of a multisubunit receptor complex result in an identical human disease phenotype. PMID- 12080487 TI - [Clinico-epidemiological study of neurolisteriosis in the province of Segovia]. AB - INTRODUCTION: Listeria monocytogenes is a microorganism with marked tropism towards the central nervous system. One of the commonest forms of presentation is as a meningeal infection. The objective of this study is to describe the epidemiological, clinical and laboratory characteristics and therapeutic implications. PATIENTS AND METHODS: We made a retrospective analysis of four patients, admitted to hospital with the diagnosis of meningeal infection and positive culture for L. monocytogenes in the cerebrospinal fluid (CSF), who were admitted to our centre between 1 January 1996 and 31 December 2000. The overall rate of meningitis due to L. monocytogenes in the province of Segovia is similar to that reported from the USA and Europe (six cases per million inhabitants per year). All the patients were male, aged between 19 and 79 years. The commonest form of presentation was a meningeal syndrome. Analysis of the CSF showed pleocytosis with high protein and low glucose levels. Gram staining was negative in all cases. Initial empirical treatment proved to be correct in two of the four patients. The only predisposing factors observed were alcoholism and diabetes mellitus. All four cases recovered. CONCLUSION: We emphasize the difficulty in diagnosing meningitis due to L. monocytogenes, since the presenting symptoms are relatively non specific, CSF analysis gives variable results and Gram staining is of little use. Clinical suspicion is important so that specific antibiotic treatment may be given to improve the prognosis. PMID- 12080488 TI - [Dementia with frontal lobe atrophy: clinical study from 18 patients with cognitive impairment and frontal lobe involvement detected by structural and functional neuroimaging]. AB - OBJECTIVE: To characterize clinically patients with cognitive impairment and frontal lobe degeneration at neuroimaging. PATIENTS AND METHODS: Patients diagnosed of dementia or mild neurocognitive impairment by DSM IV criteria and neuropsychological tests with frontal lobe atrophy and hipoperfusion detected by computed tomography and single photon emission computed tomography. RESULTS: 18 patients, 5:1 on behalf of women; mean age at onset, 74 years; hereditary for dementia, 38%; mean duration of illness at first testing, 2 years; the most common initial symptoms, memory loss. At initial examination, psychotic or behavioral impairment was found in 61% and parkinsonism in 38%, the commonest cognitive troubles at this moment was the executive function, language and anterograde memory dysfunctions at the same rate. Frontal lobe atrophy and hypoperfusion was found in 100%, temporal in 88% and parietal in 38%. Electroencephalogram was pathological in 33%. The course of the disease was progressive but with fluctuation in 27%. Frontotemporal dementia clinical criteria in 13 patients, 4 of them clinical criteria of dementia with Lewy bodies too, and clinical criteria of Alzheimer s disease in 5. PMID- 12080489 TI - [Clinical and surgical aspects of the meningiomas of the base of the skull. III. Meningiomas of the posterior fossa]. AB - INTRODUCTION AND OBJECTIVE: The close relationship between the meninges of the posterior fossa and major vascular structures, the possible involvement of many cranial nerves and proximity of the brainstem means that surgery of lesions in this region is difficult. Minimal differences in the site of origin of the tumour and its anatomical relations may affect the surgical approach and postoperative results. In this study three basic sites were distinguished (clival and petroclival, petrous or pontocerellar angle and foramen magnum meningiomas). Significant differences were observed in the three types. PATIENTS AND METHODS: We present 26 successive patients whose meningiomas of the posterior fossa were resected and the results obtained. RESULTS AND CONCLUSIONS: 1. The patients operated on included 24 women and 2 men. 2. Age was not decisive when deciding on surgery. 3. The average time between onset of symptoms and diagnosis was 14 months. 4. The petroclival meningiomas were the most difficult to excise. 5. The size of the tumours worsened the preoperative clinical situation and postoperative morbidity. PMID- 12080490 TI - [Implication of the neuropeptides methionine enkephalin, neurotensin and somatostatin of the caudal trigeminal nucleus in the experimental migraine]. AB - INTRODUCTION: Primary peptidergic sensory neurons of the trigeminal ganglion that innervate the cerebral dura have been involved in the pathogenesis of headache, including the migraine. In addition, it is known that nociceptive central processes of the trigeminal neurons terminate in the caudal trigeminal nucleus. Moreover, the electrical stimulation of the trigeminal ganglion has been used as an experimental model in order to study the vascular headache, including the migraine. AIM: To study whether there is or not a decrease of the immunoreactivity for methionine enkephalin, somatostatin and neurotensin in the caudal trigeminal nucleus after electrical stimulation of the trigeminal ganglion. MATERIAL AND METHODS: The trigeminal ganglia of Wistar albino rats of both sexes were electrically stimulated (frequency, 5 Hz; duration, 5 ms; intensity, 0,8 1.4 mA) and unilaterally for five minutes. Sections of the medulla oblongata containing the caudal trigeminal nucleus were obtained and processed for immunocytochemistry, in which specific antibodies were used against methionine enkephalin, neurotensin and somatostatin 28. RESULTS: In stimulated animals, we observed a decrease in the immunoreactivity for the three neuropeptides studied in the stimulated (ipsilateral) side, in comparison with the not stimulated side (contralateral). In control animals (not stimulated) the degree of the immunoreactivity was the same on both sides. CONCLUSIONS: 1. The decrease of the immunoreactivity in the ipsilateral side (stimulated) suggests that methionine enkephalin, neurotensin and somatostatin 28 are released in the caudal trigeminal nucleus after electrical stimulation of the trigeminal ganglion; 2. Methionine enkephalin and somatostatin 28 could act in the caudal trigeminal nucleus as inhibitors (with antinociceptive action) of another released exciters neuropeptides (with nociceptive action); and 3. These data will allow in the future to try new therapeutic strategies (e.g., the inhibition of the receptors implicated.), in order to alleviate certain headaches. PMID- 12080491 TI - [Therapeutic attitude, evolution and prognosis of the cerebral hemorrhages of patients in dicumarinic treatment, in function of their supra or infratentorial localization]. AB - INTRODUCTION: The anticoagulants are drugs with possible serious secondary effects, being one of the most serious the appearance of intracraneal hemorrhages, being able to vary the clinical course as they were hemorrhages in the deep supratentorial compartment (ST) or in the infratentorial (IT). OBJECTIVES: Compare the therapeutic attitude, the evolution and the prognostic of the cerebral hemorrhages in patient with warfarin treatment, in function of their localization ST or IT. PATIENTS AND METHODS: Descriptive and retrospective analysis of 42 patients; in 28 of the cases (66,6%) the localization was ST, and in 14 of the patients (33,3%) the localization was IT. We study risk vascular factors, age and the sex of the patients, the volume of the hemorrhage, the initial clinical state of the patients, the therapeutic attitude, the average stay and the mortality. As statistical tools, the t of Student and the c2 test were used, demanding a confidence interval of 95%. RESULTS: The factors of cardiovascular risk of our serie were similar to the existent ones in other studies. The hemorrhages IT are more frequent in women, existing significant differences in the age among sexes in function of the localization. 100% of the hemorrhages IT exceeded the therapeutic INR, in front of 39% of the ST. A bigger half stay is also demonstrated in the ST hemorrhages so much in the surviving as in the exitus. Statistical significance was not reached in the rest of the studied parameters. CONCLUSIONS: The presence of a INR that surpasses the therapeutic is an indicator of hemorrhage risk IT, in patient dicumarinic drawees, and that in this same subgrup, the stays stockings are sensibly inferior, in all the subgrups (medical treatment and surgical treatment). Also the feminine sex has a bigger incidence of processes ST, with a smaller presentation age. PMID- 12080492 TI - [Prevalence and characteristics of multiple sclerosis in the health district of the Marina Alta]. AB - OBJECTIVES: To determine the prevalence and characteristics of multiple sclerosis (EM) in the Marina Alta health district. PATIENTS AND METHODS: A descriptive, retrospective study localizing cases (crossed register method). Day of prevalence: 1/05/01. Registers of the area analyzed: hospital admission, Neurology Clinic, Emergency Department of the hospital and Health Centres. The criteria of Poser were used for diagnosis. We analyzed the demographic, clinical and paraclinical aspects of the episodes and treatment (interferon-beta). RESULTS: Male/female ratio: 2.4. Average age: 46.6 years. Prevalence of definite EM: 40.3/100,000. Prevalence of autochthonous cases of EM: 28.7/100,000. Average age at onset: 32.6 years. Average duration of illness: 12.5 years. Commonest clinical form: remitting relapsing (48.1%), followed by progressive relapsing (7%), benign (5.5%) and primary progressive (1.8%). 5.5% died, all with the progressive secondary form. EDSS disability: greater in the progressive secondary form. Magnetic resonance (MR) was the commonest investigation used (84% of the results in concordance and 13% normal). Positive BOC in 70% of cases. We studied 202 episodes. The systems most affected were: sensory and pyramidal. Worsening of EDSS after recovery from the episode: 51.7%. 29% received interferon-beta, with five cases (31%) of major side effects (one psychotic outbursts, three with neutropenia and one generalized allergic reaction). CONCLUSIONS: In the Marina Alta the prevalence of EM adjusted for the autochthonous population is in the medium risk zone. The clinical features and course of the disease in our patients are similar to those seen in other national and foreign series. The most worthwhile tests were cranial MR and BOC CSF. Treatment with interferon-beta requires strict control in view of the high frequency of serious side effects seen. PMID- 12080494 TI - [Lipoma of the cerebral convexity and refractory focal epilepsy]. AB - INTRODUCTION: Intracranial lipomas make up approximately 0.1% of all cerebral tumours. They are considered to be congenital malformations, caused by poor differentiation and abnormal persistence of the primitive meninges, which has become adipose tissue during the development of the subarachnoid space. They are usually found in the midline, often in the cisterna pericallosa. It is very unusual for them to be in the fissure of Sylvius and cerebral convexity. Most cerebral lipomas are aymptomatic. Epileptic seizures are the commonest symptom, and are sometimes refractory to treatment. CASE REPORT: A 20 year old man with no previous clinical history. From the age of three years he had frequent senso motor seizures of the lower half of the right side of his body, which were resistant to anti-epileptic drugs. On magnetic resonance there was an extra axial lesion in the left posterior parietal and parasagittal convexity. This was hyperintense in T(1) sequences, not seen in STIR sequences, and was compatible with fatty tissue. Subtotal removal of the tumour was carried out and the histopathological diagnosis was lipoma. The patient had no seizures in the year after operation. CONCLUSIONS: Lipomas of the cerebral hemispheres are very uncommon. The usual symptoms are focal epileptic seizures, as a result of the cortical irritation caused by close adherence of the lipoma to the cerebral cortex or associated cortical or vascular dysplasias. They have easily recognized characteristics on computerized axial tomography and magnetic resonance. Treatment should be conservative unless it is associated with refractory epilepsy, in which case careful resection may be considered, although this is rarely complete. PMID- 12080493 TI - [Cognitive effects of therapy with topiramate in patients with refractory partial epilepsy]. AB - INTRODUCTION: Multiple studies have demonstrated that antiepileptic drug therapy may have consequences on cognitive function. Topiramate (TPM) is a new antiepileptic drug characterized by several mechanisms of action, which has proved to be clinically effective in the reduction of the quantity of crisis in epileptic patients. Clinical studies frequently report subjective cognitive complains, inconsistently sustained. The main objective of this study was to assess neuropsychological effects attributable to the addition of TPM to the current antiepileptic therapy, in patients with refractory partial epilepsy. PATIENTS AND METHODS: 44 patients were assessed through a cognitive battery applied before beginning of therapy with TPM and 6 months after the dosage had been stabilized. Tests had been chosen to assess verbal and non verbal memory, executive functions, language, visuomotor processing and psychomotor functions. The cognitive study was conducted in parallel with clinical trials TOPMAT-EPAJ 111 and TOP-POR1. RESULTS: The statistical analysis of results didn t show changes in motor tests, in sustained attention and in logical and visual memory. Deterioration of semantic verbal fluency, verbal learning, work memory and visuomotor skills was observed. Considering the effects of the dosage of topiramate and the total quantity of antiepileptic drugs, major commitment was observed in patients taking more than 400 mg/day. CONCLUSIONS: Like other antiepileptic drugs, topiramate, used in polytherapy in patients with refractory epilepsy, may have consequences on cognitive functions. These changes may be related with the potentiating action of this drug at the level of neurotransmission system of gamma amino butyric acid (GABA), substance that has inhibitory properties in the fore regions of the brain. PMID- 12080495 TI - [Wilson's disease: a clinical case is presented with neuropsychiatric dominant form and probable physiopathology interpretation with magnetic resonance of the brain]. AB - CASE REPORT: A clinical case of Wilson's disease is presented. She was a 26 years old woman who began to show psychological symptoms, and later developed neurological signs such as asymmetrical hand tremor, parkinsonism, dystonia and later dysphagia and mutism. The ophthalmological examination found a Kayser Fleischer ring in the Descemet membrane. There was disturbance of copper metabolism documented with reduction of serum ceruloplasmin and increase of the urinary excretion of copper. Cirrhosis was demonstrated through laparoscopy and liver biopsy. RESULTS: The brain magnetic resonance showed frontotemporal atrophy and a degenerative process at the basal ganglia, cerebellum and brain stem, data which could de used to suggest the probable neuropsychiatric physiopathology. The stenosis and intense cervical dysphagia, associated with crycopharyngeal membrane, has not been mentioned previously. PMID- 12080496 TI - [Calcification of the thoracic yellow ligament as a cause of myelopathy]. AB - INTRODUCTION: Ossified yellow ligaments are a rare cause of thoracic myelopathy, which has been reported mainly in Japanese patients. The lower thoracic spine is the most common site affected and symptomatic patients usually begin to develop spactic paraparesis in combination with reduced depth sensation. CLINICAL CASE: A 45 year old female that since 8 months presented progressive gait disturbance with paresthesias to both lower extremities. Neurologic examination revealed a spactic gait with exaggeration of lower extremities reflexes, positive bilateral Babinski sign and reduced depth sensation. The MRI revealed concentric narrowing of the spinal canal between D(8) to D(11) with spinal cord compression due to enlarged calcified yellow ligaments and hypertrophy of articular processes. A D(9)-D(10) laminectomy was performed, with removed of calcified yellow ligaments, and foraminotomy in affected levels. The postoperative follow up during 30 months has been uneventful. CONCLUSION: Calcified yellow ligaments are a poorly understood cause of thoracic myelopathy which should be treated as early as possible in symptomatic patients. PMID- 12080497 TI - [Paralysis of cranial nerves as a form of presentation of dissection of the internal carotid artery]. AB - INTRODUCTION: Dissection of the internal carotid artery (DIC) is a known cause of cerebral infarct, especially in young patients. The classical clinical syndrome consists of unilateral pain of the head or neck, homolateral oculo-sympathetic paresis and ischaemic symptoms of the cerebral hemisphere involved. Presentation as paralysis of cranial nerves is rare and occurs in less than 12% of cases. The neurological involvement seems to be due to compression caused by the increased diameter of the artery involved. CASE REPORTS: Two patients are reported with paralysis of the lower cranial nerves secondary to DIC. In the first case there was paralysis of the left cranial nerves IX, X, and XII which was diagnosed on angiography using computerized tomography with spiral acquisition. The second patient had clinical involvement of cranial nerves IX, X, XI and XII and magnetic resonance angiography showed the dissection. Both cases were confirmed after digital subtraction angiography. CONCLUSION: Diagnosis of DIC requires a high level of suspicion in cases with atypical onset. The use of new techniques of non invasive imaging diagnosis such as computerized tomography and magnetic resonance angiography permit effective diagnosis of this disorder. PMID- 12080498 TI - [Structure of the cerebral cortex. Intrinsic organization and comparative analysis of the neocortex]. AB - We review our present knowledge on the intrinsic organization of the neocortex based on studies carried out with the Golgi method in several mammalian species. An outline is presented on certain general aspects of the termination of specific afferent fibers in layer IV in insectivora, rodents, carnivora and primates. The principal components of the cerebral cortex have been classified in two broad types: pyramidal cells, which account for nearly 70% of the total population, and intrinsic neurons. We review different morphological characteristics of pyramidal cells and intrinsic neurons with a full description of cell varieties, dendritic morphology and several aspects of synaptic connectivity of intrinsic neurons in comparison between different species. Special attention has been paid to the description of spinous stellate cells which seems unique for a given animal species. We have reviewed intrinsic cortical circuits linking several neocortical layers, the distribution of dendritic plexuses, and the excitatory and inhibitory synaptic connections established between various neuronal categories. We finally review modern concepts of functional neocortical architecture based on the modular organization of the cerebral cortex, concluding that the cerebral cortex is rather uniform at en elementary level but the differences appear substantial when the comparison is made between different species. PMID- 12080499 TI - [Dementia associated with AIDS]. AB - INTRODUCTION: Dementia associated with AIDS or AIDS dementia complex (ADC) is characterized by neurocognitive defects with disorders of the functional ability of the patient for work and the activities of everyday life. The increasing number of AIDS patients has led to a significant rise in those with neurological involvement, especially in poorer countries. DEVELOPMENT: At present no data on the incidence of ADC is available. The prevalence varies with the population studied and criteria used for diagnosis. Some authors have suggested a frequency of 8 to 16%. However, other reports mention an incidence of up to 20.7% of encephalopathy in AIDS patients. The mechanism by which ADC occurs is still not clear. Many factors have been suggested but none confirmed as causal. CLINICAL FEATURES: symptoms are due to subcortical dementia with difficulty in concentration, memory and learning whilst language, recognition, basic attention and executive function are relatively well preserved. DIAGNOSIS: this is essentially clinical, by exclusion, in cases of HIV+ patients with marked immunological deterioration and compatible clinical features. TREATMENT: current clinical guidelines recommend that most AIDS/HIV patients, if not all, should receive treatment with potent combinations of anti retroviral drugs. CONCLUSION: Even in the age of modern, highly active anti retroviral treatment, ADC is still a devastating complication of HIV infection. PMID- 12080500 TI - [Considerations about late-onset epilepsy]. AB - OBJECTIVE: To make a revision of the general point of view on late-onset epilepsy, known as epilepsy that starts in the mature age after 25 years old; around 25% of the patients with epilepsy had their first crisis after that age, with an increase in the incidence in the course of the age. DEVELOPMENT: The main ethiologies are discussed, standing out: alcoholism (22%), stroke (18%), tumors (10%), metabolic imbalances (10%), infections of the CNS, trauma, atrophies and cisticercosis. For the diagnosis is required a complete clinical evaluation, cardiovascular examination, metabolic tests, EEG and neuroimage studies. CONCLUSIONS: Monotherapy with phenobarbital, carbamazepine and valproate, control 80% of cases and failure related to patients with wide spread cerebral damage. PMID- 12080501 TI - [An epidemiological study about ependymomas in Aragon and La Rioja]. PMID- 12080503 TI - [An epidemiological study about gliosarcomas in Aragon and La Rioja]. PMID- 12080502 TI - [An epidemiological study about cerebral epidermoid cysts in Aragon and La Rioja]. PMID- 12080504 TI - [An epidemiological study about craniopharyngiomas in Aragon and La Rioja]. PMID- 12080505 TI - [An epidemiological study of hypophyseal adenomas in Aragon and La Rioja]. PMID- 12080506 TI - [An epidemiological study of astrocytomas in Aragon and La Rioja]. PMID- 12080508 TI - [Functional magnetic resonance in the evaluation of the activity of the anterior cingulate cortex using Stroop's paradigm]. AB - INTRODUCTION: Stroop s paradigm has been used to evaluate the anterior attention system which regulates the inhibitory capacity of automatic responses. Functional neuroimaging techniques have shown a preponderant role for the anterior cingulate cortex in carrying out this paradigm. OBJECTIVE: To evaluate the activity of the anterior cingulate cortex in view of its clinical importance in the study of neurological and psychiatric disorders. PATIENTS AND METHODS: Eleven healthy volunteers took part in the study. The functional images were analyzed using the software SPM99 and by second order individual and group analysis. RESULTS: Initial local analysis showed activation in the right anterior cingulated cortex (Brodmann s area 32) and left central (areas 31 and 23); caudate nucleus (right body and left tail) and thalamus (bilateral). Overall there was significant activation of the left hemisphere, in areas 44 (Broca s area), 7, 40 (supra marginal gyrus and insular cortex, and in the right hemisphere in area 19. In spite of this there was great individual variation. CONCLUSIONS: The overall results are concordant with complex functional connections for attention and the control of automatic responses. In our study the anterior cingulated cortex was not selectively activated. The activation of the thalamus and caudate nucleus may be explained by their involvement in the frontostriatal circuits. The lack of individual consistency may be due to different personal cognitive styles of resolving conflicts. According to our results, Stroop s paradigm would not be clinically useful for showing good or bad functioning of the anterior cingulated cortex. PMID- 12080507 TI - [Carer's burden and depressive symptoms in patients with Alzheimer s disease. State after twelve months]. AB - OBJECTIVE: The objective of this study is to determine the effect of depressive symptoms of patients in the burden perceived by the carer, evaluate the course of this burden in relation to the disappearance, persistence or appearance of depressive symptoms after twelve months and find which factors are associated with a rise or fall in the carer s load after twelve months. PATIENTS AND METHODS: A prospective observational study for 12 months of a sample of 150 patients with the clinical diagnosis of probable Alzheimer s disease on the criteria of NINDCS ADDRA: The sociodemographic data of their carers were collected and tested on the following scales: CAMDEX, CAMCOG, NPI, RDRS 2 and Burden Interview (BI) initially and twelve months later. RESULTS: The existence of depression initially was associated with an increased burden (score of BI of 22.76 compared with 15.79), increased non cognitive symptoms (NPI score of 16.23 compared with 5.94) and greater functional disability (RDRS 2 score of 27.89 compared with 25.53). The burden increased in the group of patients whose depressive symptoms persisted and in those in whom they appeared after twelve months. CONCLUSION: The data suggest that depressive symptoms increase functional disability and are associated with an increase in non cognitive symptoms. At the same time the functional disability and non cognitive symptoms increase the carer's burden. PMID- 12080509 TI - [The evolution of use of anti-Parkinson drugs in Spain]. AB - INTRODUCTION: In recent years new anti Parkinson drugs have been marketed and there has been controversy over the safety of some drugs. OBJECTIVE: To analyze the evolution of the consumption of anti Parkinson drugs and the effect of the newer drugs. PATIENTS AND METHODS: A study of the consumption of anti Parkinson drugs (1989 1998). Data were obtained from the ECOM database of the Ministry of Health and TEMPUS of the National Statistics Institute. The drugs were classified using the Anatomo Therapeutic Clinical Classification (ATC). Consumption was expressed in defined daily dosage (DDD) and the costs in euros. The drugs marketed since 1990 were classified as new drugs and the others as classical drugs. RESULTS: The total consumption of drugs increased from 1.92 DDD/1,000 inhabitants/day in 1989 to 3.64 DDD/1,000 inhabitants/day in 1998. The drugs showing the greatest increase were selegiline, pergolide and levodopa. The total pharmaceutical expenses tripled. There was a smaller increase in the consumption of new drugs (1.2% of the total in 1991 and 6.6% in 1998) than in their costs (6.7% of the total in 1991 and 38.8% in 1998). The cost per DDD of the new drugs increased five times (1989: 2.55 euros and 1998: 13.59 euros) and that of the classical drugs was similar (1989: 0.54 euros and 1998: 0.62 euros). CONCLUSIONS: The total consumption of anti Parkinson drugs has progressively increased. The consumption of selegiline has also increased in spite of controversy over its safety. The new drugs have a major economic effect. PMID- 12080510 TI - [Clinical features of cerebral toxoplasmosis in Cuban patients with AIDS]. AB - INTRODUCTION: Neurotoxoplasmosis (NTX) is one of the commonest opportunist infections in patients infected by the human immunodeficiency virus. It presents with a variety of clinical features in the immunocompromised patient. PATIENTS AND METHODS: We made a horizontal retrospective descriptive study of 88 patients with a diagnosis of NTX treated in the Pedro Kour Institute during a three years period. Our aim was to find which were the most common forms of clinical presentation in our setting and to evaluate the use of paraclinical examinations in reaching the presumptive diagnosis. The patients had clinical examinations, CSF studies, detection of indirect immunofluorescence titres (IFI) for Toxoplasma, imaging studies (CAT) and lymphocyte counts. RESULTS: The commonest symptoms were: headache in 79%, fever in 55.68%, motor defect in 44.31% and disorders of consciousness in 29.54% of the patients. The IFI titres were negative in 31.54% of the patients, between 1/16 and 1/32 in 38.34% and between 1/64 and 1/1,024 in only 30.07% of cases. The T CD4+ lymphocyte count was less than 200 cells in 66.7% of the patients; 72% of imaging studies showed typical lesions, in association with other disorders in 38% of the cases (lymphomas, cryptococcosis, tuberculous meningoencephalitis and cytomegalovirus encephalitis). CONCLUSIONS: NTX in a patient with AIDS often presents in our setting with headache and fever, motor deficit and alterations of consciousness. The diagnosis should be confirmed by immunological or imaging studies since in this disease serology and CSF studies are less specific. PMID- 12080511 TI - [Type A behaviour and life events as risk factors for cerebral infarct]. AB - INTRODUCTION: The role of psychosocial factors in cerebral infarct has been little studied and is therefore still undefined. OBJECTIVE and methods. To evaluate the effect of a pattern of behaviour which predisposes to stress, and of life events, on the occurrence of cerebral infarct, we studied a group and control involving 88 patients with cerebral infarcts diagnosed clinically and on imaging studies, and a control group of 99 persons paired for age and sex who lived in our health district. To evaluate the behaviour pattern we used Jenkins abbreviated questionnaire and for life events the Holmes and Rahe questionnaire. RESULTS: The type A behaviour pattern had a highly statistically significant association with cerebral infarct, which was not associated with the effects of other variables. This relation was the greater the lower the age of the person. It was not seen after the age of 75 years. The behaviour pattern was not related to arterial hypertension or with the aetiopathological mechanisms of the patients. In relation to the life events questionnaire, the number of events was significantly associated in the group of patients, but not with the total score of the questionnaire. However, the latter was significantly related to hypertensive patients and persons with a type A behaviour pattern. The commonest life events in both groups were related to the death or health problems of family members, the previous illnesses of the person and to financial and work problems. CONCLUSION: The type A behaviour pattern is individually associated with cerebral infarct, whilst stressful life events were involved in the cerebral infarcts of hypertensive and type A behaviour patients. PMID- 12080512 TI - [Clinical and surgical aspects of meningiomas at the base of the skull. II. Meningiomas of the middle fossa]. AB - INTRODUCTION and objective. In spite of the advances made over the past two decades in neuro anesthesia, neuro radiology and neurosurgical microsurgery, meningiomas of the base of the skull are still a challenge in the field of neurosurgery. In this paper we describe the clinical and surgical characteristics of meningiomas of the middle fossa and our experience in their surgical treatment. PATIENTS AND METHODS. We describe 24 patients who were operated on consecutively for meningiomas of the temporal fossa, lesser wing of the sphenoid and cavernous sinus and the surgical results obtained. RESULTS and conclusions. The results obtained included: 1. Predominance of women for all sites. 2. Age was not a decisive factor when deciding whether to operate or not. 3. The average period between onset of symptoms and diagnosis was two years. 4. In most cases resection of the tumour was total, as confirmed on macroscopy studies. 5. There was a significant increase on the Kamofsky scale postoperatively whatever the site of the tumour resected. 6. The tumours of the cavernous sinus were technically the most difficult to resect surgically. 7. The size of the tumour worsened the preoperative clinical condition and postoperative morbidity. PMID- 12080513 TI - [Examination of the hypothesis 'the factors and mechanisms of superiority']. AB - INTRODUCTION. The hypothesis of Geschwind and Galaburda suggests that specific cognitive superiority arises as a result of an alteration in development of the nervous system. In this article we review the co existence of superiority and inferiority . PATIENTS AND METHODS. A study was made of six children aged between 6 and 8 years old at the Instituto de Belles Artes Antonio Maria Valencia in Cali,Columbia with an educational level between second and third grade at a primary school and of medium low socio economic status. The children were considered to have superior musical ability by music experts, which is the way in which the concept of superiority was to be tested. The concept of inferiority was tested by neuropsychological tests = 1.5 DE below normal for the same age. We estimated the perinatal neurological risk in each case. Subsequently the children s general intelligence and specific cognitive abilities were evaluated. In the first case the WISC R and MSCA were used. The neuropsychological profiles were obtained by broad evaluation using a verbal fluency test, a test using counters, Boston vocabulary test, the Wechster memory scale, sequential verbal memory test, super imposed figures test, Piaget Head battery, Rey Osterrieth complex figure and the Wisconsin card classification test. The RESULTS showed slight/moderate deficits in practical construction ability and mild defects of memory and concept abilities. In general the results supported the hypothesis tested. The mechanisms of superiority proposed in the classical hypothesis mainly involve the contralateral hemisphere: in this study the ipsilateral mechanism was more important. PMID- 12080514 TI - [The association between post anoxic encephalopathy and startle epilepsy]. AB - INTRODUCTION: The startle is a variant of primitive physiological reflex, present in developed animals. It could be defined as a sudden defensive reaction triggered by an adequate sensorial stimulus, most effective if auditory, consisting under a clinical basis on the sudden and sequential contraction of some muscular groups, in first place affecting the facial territory (orbicularis oculi bilaterally), followed by the rest of the cranial nerves, the axial musculature and finally the limbs, predominantly proximal and flexor regions of superior extremities. The pathological startle is that which is precipitated by inadequate stimulus and excessive in response to it (under a low level of intensity or duration). It appears on account of different causes and mechanisms, among them a peculiar form of reflex epilepsy, the startle epilepsy. This form of epilepsy commonly present on infant postanoxic encephalopathy, shares its clinical features with other causes of pathologic startle, even though it presents a very characteristic electroencephalographic semiology. CASE REPORT: We present the case of a child with a startle epilepsy clinically presented as a form of subintrant status epilepticus under the context of a postanoxic encephalopathy secondary to drowsiness. With an adequate treatment it was possible to eliminate the epileptic phenomena, thus notoriously improving his quality of life (vigil and asleep). CONCLUSION: We review the characteristics and classifications of a variant of reflex epilepsy, the startle epilepsy. PMID- 12080516 TI - [Fusiform aneurysm of the middle cerebral artery]. AB - Introduction. The main objective of the treatment of intracranial aneurysms is to isolate them from the cerebral blood circulation. A fusiform aneurysm, because of its shape, cannot be treated using the usual techniques and usually requires techniques of arterial reconstruction and revascularization using by pass. Currently it is possible to find the vascular territories with the greatest risk of causing neurological defects and where revascularization is necessary. CASE REPORT. A 20 year old man with no previous history of illness who had several transient ischaemic episodes. Emergency laboratory tests, ECG and plain chest Xray were all normal. Imaging investigations showed the presence of a fusiform aneurysm of the anterosuperior division of the right middle cerebral artery. No associated systemic disease was detected. Wada s test showed the vascular territory with the greatest risk of neurological deficit. Extra intra cranial by pass was done from the right superficial temporal artery to the distal portion of the anteriorsuperior branch of this artery. The operation was done using a right pterion approach with dissection of the superficial temporal artery, and the aneurysm, trapping and termino lateral anastomosis. Cerebral protectors and mild hypothermia were used during the operation. The post operative course was uneventful. Anatomo pathological diagnosis was of an atherosclerotic fusiform aneurysm with osseous and chondroid metaplasia. After six months follow up the patient remains asymptomatic. Discussion and conclusions. Treatment of fusiform cerebral aneurysms is complex and usually requires procedures for cerebral revascularization. Correct pre operative evaluation is essential to identify the vascular territory with the greatest risk of causing neurological deficit. Wada s test is useful for this, since it permits selective evaluation of the different vascular territories safely and quickly. High or low flow by pass of these territories prevents cerebral ischaemia and permits the treatment of choice for these aneurysms. PMID- 12080515 TI - [Cutaneous haemangioma associated with a hepatic haemangioma and neurofibromatosis type 1 (NF1)]. AB - OBJECTIVE: To present a girl with cutaneous cavernous hemangioma associated with hemangioma of the liver and neurofibromatosis type 1 (NF1). PATIENTS AND METHODS: A 3 year old girl showed cutaneous cavernous hemangioma on the right hypochondral region and numerous caf au lait spots. She was studied with abdominal echography and abdominal and thoracic magnetic resonance (MR) as well as with cerebral MR angiography. RESULTS: Both image studies, echography and MR, revealed a mass 2 x 2 cm of hemangiomatous type in the upper zone of the liver, near the external cutaneous cavernous hemangioma. Echographical follow up showed a progressive diminution of the hepatic hemangioma size. At 2 1/2 years of age a voluminous thoracic tumor extending between aorta and sternon very difficult of being removed was found. CONCLUSION: Hemangioma of the liver appears to be related with cutaneous cavernous hemangioma as a finding of the syndrome of Pascual Castroviejo II. Furthermore, the patient has NF1 which has not been previously related with the cutaneous and/or the liver pathology. PMID- 12080517 TI - [Neurobiological basis of drug addiction]. AB - INTRODUCTION. Drugs of abuse cause addiction, characterized by compulsive and out of control drug use. Several neurochemical and molecular changes take place in the brain during the first phase of drug use which lead to dependence, addiction and drug abuse. DEVELOPMENT. Every drug of abuse acts through similar neurophysiological mechanisms, mostly based on the abnormal activation of the mesolimbic dopamine system. Although the addicted brain tries to adapted to the drug action, there are permanent neural changes at the cellular and molecular levels which seem to underlie the addictive process and the emergence of abstinence symptoms after drug taking stops. Furthermore, these neural changes appear to subserve the emergence of cravings once the abstinent syndrome has been overcome. Cravings are the main factor leading to relapse, and they are often initiated by exposure to drug related cues. PMID- 12080518 TI - [Chronic hydrocephaly in adults. A diagnostic and therapeutic challenge]. AB - OBJECTIVE: To describe the main clinical features of the different conditions which present as chronic hydrocephaly, emphasising the diagnosis and treatment. DEVELOPMENT: In the study of any dementia syndrome, normotensive hydrocephaly (HN) is of particular interest since this condition may be treated and cured. However, prediction of which cases will benefit from surgery is one of the most important aspects of diagnosis. The development of paraclinical investigations of the dynamics of cerebrospinal fluid (CSF) has led to great advances in the management of these chronic forms of hydrocephaly, since they are the best parameters for prognosis when deciding whether to insert a by pass. The treatment of post traumatic hydrocephaly has also benefited from these studies, since the cases of simple atrophy may now be distinguished from those which follow a clinical course similar to HN. Recently, two types of hydrocephaly have been identified as being very different from the typical forms. These are external hydrocephaly, in which CSF accumulates in the convexity of the cerebral hemispheres, and long term ventriculomegaly, in which severe ventricular dilation is accompanied by cognitive alterations and features of acute hydrocephaly. CONCLUSIONS: Treatment of each of these conditions is a challenge. Therefore careful analysis of the clinical features, imaging studies and CSF dynamics is essential when deciding whether or not to use by pass treatment. PMID- 12080519 TI - [Executive functions: the need for the integration of concepts]. AB - INTRODUCTION: The new cognitive neuropsychology approaches have aroused an increasing interest in understanding the higher cognitive processes as well as the neural substrates linked to them. Particularly, the executive functions, reckoned to be essential to control the information processing and to co ordinate behaviour, have received preferential treatment from specialised literature on the subject. DEVELOPMENT: From obsessive compulsive disorder to schizophrenia, from Parkinson s disease to multiple sclerosis, there are many reports that show the affectation of these functions in all these morbid processes. On the other hand, the part that the prefrontal cortex plays in human behaviour in general, and in executive functions in particular, constitutes one of the most important fields of research of neurosciences nowadays. Thus, this cortical area appears closely linked to the executive processes, affecting different respects of the cognitive functions. Working memory, supervisory attentional system, somatic marker, information processing, behaviour planning, social judgement, are processes which have been related to the prefrontal cortex activity as a structure, and to the executive processes as a function. CONCLUSIONS: The aim of this article is to revise the concept of executive functions, and give rise reflections about the usefulness of the aforementioned concept and its practical applicability. It is essential that we understand the difference between structure and function, cognition and emotion, brain activity and behaviour, category and dimension, and between mind and brain, to achieve a more comprehensive approach to this concept of executive functions we all use, and many times find difficult to define and to understand. PMID- 12080520 TI - [Parkinson's disease and smoking: coherence and plausibility]. AB - INTRODUCTION: Many studies have shown that smoking is lower in patients with Parkinson s disease. However, in other investigations this was not observed. The various studies involved showed wide variation with regard to methodology, criteria for diagnosis and periods of observation and hence it is difficult to compare them. DEVELOPMENT: The first studies published were designed to examine the effects of smoking in general and information was obtained regarding the possible disorders related to tobacco smoking according to the records of mortality, which may contain errors due to selective mortality and mistaken diagnosis. Most of the studies of cases and controls included prevalent cases which accepted the study, mainly hospital cases. Also it is probable that the prevalent cases of Parkinson s disease do not smoke because of their disorders of movement. CONCLUSIONS: Many researchers have found important information about the pathophysiology of Parkinson s disease and its association with smoking. However, the hypothesis regarding the association between smoking and low risk of Parkinson s disease are various and independent, apart from the hypothesis of a truly biological mechanisms. Since the subject is still controversial, systematic reviews together with epidemiological and experimental studies are necessary. PMID- 12080522 TI - [Neuropathy of the superficial peroneal right nerve in the region of the ankle]. PMID- 12080523 TI - [Frontal lobe dementia of vascular aetiology. A case report]. PMID- 12080524 TI - [Rett's syndrome: clinical description and differential diagnosis]. PMID- 12080527 TI - Professional nurses rise above the call of service. PMID- 12080525 TI - [A reply to: Babinski reflex, yes; Babinski sign, no; Pseudobabinski never!]. PMID- 12080528 TI - Workplace advocacy = patient advocacy. PMID- 12080529 TI - EMTALA and the nurse. PMID- 12080535 TI - Bioterroism. PMID- 12080532 TI - Terrorism response resources for clinicians. PMID- 12080536 TI - The oncology nurse's role in educating patients on endocrine therapy for metastatic breast cancer--focus on fulvestrant. AB - Fulvestrant, an estrogen receptor downregulator, is an effective and a safe endocrine option for postmenopausal women with advanced breast cancer who progress or develop recurrent disease on prior endocrine therapy. The recommended dose is 250 mg given as a single 5-mL or 2 concurrent 2.5-mL monthly intramuscular injections. Common adverse events associated with fulvestrant are hot flashes, nausea, and mild injection site reactions. Applying warm or cold compresses to the injection site can minimize injection site reactions. In randomized double-blind clinical trials, the frequency of injection site reactions for fulvestrant are no different from the comparator (anastrozole) arm. Overall, fulvestrant has a safety profile similar to aromatase inhibitors. Advantages of this agent are its effectiveness in tamoxifen-resistant tumors and the lack of agonistic property, creating a favorable side-effect profile. One limitation is that there is no evidence regarding the safety and effectiveness of fulvestrant in premenopausal women. Among endocrine therapies used to treat breast cancer, fulvestrant is unique not only in its mechanism of action but also in its mode of administration. With oral therapies, the patient fills the prescription in a pharmacy and takes the medication home. In contrast, with monthly fulvestrant intramuscular injection, the patient will have increased contact with the nurse. The increased interaction between the patient and the nurse will affect the role of nurses providing patient education and monitoring. PMID- 12080537 TI - Estrogen receptor downregulators: new advances in managing advanced breast cancer. AB - Breast cancer is the most frequently diagnosed malignancy in American women. Although potentially curable in the early stage, metastatic breast cancer, however, is essentially incurable. Because metastatic breast cancer is unlikely to be cured with currently available therapies, the goals of treatment in this setting are prolonging survival and maintaining or improving the quality of life. In patients with receptor (estrogen or progesterone)-positive metastatic breast cancer, endocrine therapy is generally considered the preferred first-line therapy because of the superior tolerability and the similar efficacy when compared with chemotherapy. Today a variety of endocrine therapies are available for the treatment of metastatic breast cancer, among which tamoxifen is the most widely used. Despite its proven efficacy and safety, a significant proportion of patients do not respond to tamoxifen. Further, due to its partial estrogenic activity, tamoxifen is associated with an increased risk of thromboembolism and endometrial cancer. Agents without any estrogenic activity-estrogen receptor downregulators have therefore been developed. Fulvestrant is the first clinically proven agent of this class. Oncology nurses have a key role in managing and educating patients with breast cancer. This supplement provides nurses with an overview of existing and emerging endocrine therapies for postmenopausal patients with metastatic or advanced breast cancer. PMID- 12080538 TI - Existing and emerging endocrine therapies for breast cancer. AB - Endocrine therapy is first-line therapy for patients with estrogen receptor positive or progesterone receptor-positive metastatic breast cancer. Commonly used endocrine therapies are tamoxifen, megestrol acetate, and aromatase inhibitors. Although tamoxifen and megestrol acetate have a favorable therapeutic profile, there are risks associated with these agents. With tamoxifen, the partial agonist property can lead to thromboembolic events. An important adverse event of megestrol acetate is weight gain and fluid retention in some patients. The aromatase inhibitors are currently used as second-line therapy after tamoxifen failure. A recent study showed that anastrozole, an aromatase inhibitor, is as effective or even superior to tamoxifen when used as a first line therapy. However, not all patients will respond to currently available therapies. A new class of drug, the estrogen receptor downregulators, has been developed. Fulvestrant, the first agent in this new class, not only induces the degradation of the estrogen receptor but also is an estrogen antagonist; further, its lack of agonist activity provides a better safety profile. Two phase III trials have proven that fulvestrant is at least as effective as anastrozole in postmenopausal women with advanced breast cancer. Fulvestrant is an effective and safe endocrine therapy for postmenopausal women who have failed prior endocrine therapy. PMID- 12080539 TI - Senate panel probes, criticizes contracting practices of GPOs. PMID- 12080540 TI - GPO or go it alone? There's a third way. PMID- 12080541 TI - Acing a survey of competence standards. PMID- 12080542 TI - Tips on meeting competence standards. PMID- 12080543 TI - Myths and facts in competence assessment. PMID- 12080544 TI - Getting MD buy-in on cost management. PMID- 12080545 TI - Deal consolidates ASC software vendors. PMID- 12080546 TI - HIPAA: a to-do list for privacy rule. PMID- 12080547 TI - Plastic surgeons take steps for safety. PMID- 12080548 TI - "Managing the mix" key for OR business. PMID- 12080549 TI - Draft guideline on processing practices. PMID- 12080550 TI - Why should nurses care about substance abuse parity? PMID- 12080551 TI - Legislative connection: summary on the nursing shortage--March 6, 2002. AB - The panel was very well received and very informative to the legislators. The nursing shortage crisis in Colorado was succinctly identified. No single solution will fix the nursing shortage. Replacement of registered nurses by unlicensed assistive personnel, or using licensed personnel with less education, will add to increased patient safety issues, provide fewer learning experiences for nursing students, and decrease job satisfaction for current nurses. If a practice that decreases quality of care is put into law as a "quick fix," that standard is very difficult to change. Legislators must listen to experienced nurses so that the decisions they make on health policy will positively influence the nursing shortage issues. PMID- 12080552 TI - [Fetal thoraco-omphalopagus. Prenatal ultrasonographic diagnosis at 15 weeks]. AB - A case of ultrasonographic prenatal diagnosis of conjoined thoraco-omphalopagus twins is reported at the fifteenth week of amenorrhea. Ultrasonography is the only mean of antepartum diagnosis of conjoined twins; Several diagnostic criteria are recalled as well as etiopathogenic and epidemiological considerations of this rare malformation which may concern 1/15.000 to 1/100.000 pregnancies. PMID- 12080553 TI - [Splenic arteriovenous fistula and portal hypertension. A case report]. AB - The authors report a rare case of portal hypertension following to an arteriovenous fistula. The embolisation of the fistula permitted to treat portal hypertension and liver histologic alterations. PMID- 12080554 TI - [Leiomyosarcoma of the penis: a case report and review of the literature]. AB - The sarcoma of the penis is a very rare tumor more exceptional is the Leiomyosarcoma of the penis for which the diagnosis required an immunohistochemical study. The prognosis for this tumor is poor. We report a case, which arise to a 53 year-old-man in this instance we did a review of the medical literature. PMID- 12080555 TI - [Assessment of cancer incidence in Tunisia 1993-1997]. AB - The objective of this study is to estimate in Tunisia on a national scale the incidence of the main cancerous localizations for the period 1993-1997 while taking results of registers of the cancer of the region of the North, Sousse and Sfax as a basis. After having verified the relation of exponential type between the incidence of cancers and the life expectancy at the birth (Evo) in a certain number of country we extrapolated levels of governorates incidence included in registers of the North, Sousse and Sfax to the other governorates while taking the similarity of the Evo level as a basis. The incidence rate of in all localizations are 100.11/100.000 for male and 86.4/100.000 for female. The main cancerous localizations for male are the lung (20.8/100.000) the bladder (10.7/100.000), the cutaneous cancers (7.2/100.000) and the prostate (6.1/100.000). For female the main localization are the breast (19.7/100.000), skin (5.8/100.000) and the cervix uteri (4.8,100.000). These results illustrate the interest of tobacco control, screening of the breast cancer of and cervix uteri cancer, as components fundamental of the cancer control. PMID- 12080556 TI - [Antigliadin antibodies in rheumatoid arthritis]. AB - We had study on the antibodies antigliadin type IgA in 40 patients having rheumatoid arthritis (33 Women and 7 men; middle age 51 years; rheumatoid factors was positive in 60%; the follow up of patient is 5.9 years). The technique applied is by ELISA. All the patients benefited of malabsorption biological test; of the immunoglobulins; and complex of histocompatibility (HLA); and gastrointestinal exploration composed of: fibroscopy duodno-jejunal with systematic biopsies. The search for this antibodies antigliadin (AAG) showed itself negative at all patients. Has histology, all these patients have a height villies normal. The abnormalities had been found in 37.5%, are minor partial intestinal villi, cellular infiltration of chorion and intraepithelial infiltration by lymphocytes more then 40%. By the way, we discuss the different mechanisms of these digestive involvment. PMID- 12080558 TI - [Congenital diaphragmatic hernias in the adult: report of six cases]. AB - The anterior hernia or the fissure of Larrey and Bochdalek hernia are exceptional in adult. We present 6 cases of congenital diaphragmatic hernia revealed in adult. Three hernia of the fissure of Larrey and three Bochdalek hernia. We discuss the different clinical, radiological, and therapeutic aspects of these hernia. We insist on the gravity of strangulated hernia and their surgical treatment. PMID- 12080557 TI - [Profile of leg ulcers. A review of a series of 70 Tunisian cases]. AB - Leg ulcer often requires hospitalization in Dermatology and involves problems of therapeutic management. We intended to draw a clear epidemiological, clinical and therapeutic profile of leg ulcer through a series of 70 cases observed over a period of 46 months in the Dermatology department at Charles Nicolle Hospital in Tunis. Like other north african works, our survey stresses a particular epidemiological profile. In fact, leg ulcer affects young men at a ratio of 5 to 1 and an average age of 43.6 years. The vascular etiological factor accounts for 63.4% of all cases of leg ulcer whereas other etiologies make up a wade range owing to their multiplicity. Yet, they may reveal a potential pathology. Treatment is both symptomatic and etiological. Treatment with saccharose has been tried on 23 patients among whom 82% showed good tolerance and 47% complete healing. As they are carried out, our results are compared with the data of literature. PMID- 12080559 TI - [Prognostic value of echocardiographic parameters in chronic cardiac insufficiency]. AB - This work was interested in 72 patients (81% males) with chronic heart insufficiency. Ail these patients had a sinusal rythme. The etiology was mainly idiopathic (36%) but also ischemic (64%). 15% of patients were at stage I of NYHA, 37.5% at NYHA II, 40.5% at NYHA III and 7% at NYHA IV. All patients had taken a medical treatment (in 85% of cases, this treatment was based on diuretics and IEC). Transthoracic echocardiography was performed in these patients to determine the telediastolic diameter (TDD = 69 mm), the telesystolic diameter (TSD = 57), the velocity of E wave (0.66 mis) and A wave (0.55 mis) and the E wave deceleration time (DT = 0.196 s), 11 from ali patients were dead after a period of observation 21 months. The factors of pejorative prognostic were: the stages III and IV of NYHA, the cardiothoracic ratio > 0.60 and the echocardiographic parameters: TDD, TSD and DT (p = 0.03, p = 0.02 and p = 0.0001 respectively). But the E and A velocity seems don't influence the prognostic. PMID- 12080560 TI - [Bronchopulmonary aspergillosis: study of 17 cases]. AB - Aspergillosis is a fungic infection depending on the local or general physiologic and immunologic state of the host. We report the result of retrospective five year study (1995-1999) about 17 cases in the laboratory of Parasitology-Mycology of Rabta hospital in Tunis. Six aspergillomas were observed, they occurred after a pulmonary tuberculosis, two cases of allergic broncho-pulmonary aspergillosis described in two asthmatic patients, nine cases of invasive pulmonary aspergillosis complicating two cancers, one leukaemia, six chronic granulomatous disease. Aspergillus fumigatus is the most frequent species (67%). The clinical and biological characteristic of those will be studied, and compared with those of the literature. PMID- 12080561 TI - [Cerebral venous thrombosis and Behcet's disease]. AB - Behcet's disease is a chronic relapsing multisystem disorder of unknown etiology. Neurological complications are frequent, occurring in 10 to 49% of cases. We report 4 cases with Behcet's disease (3 females and 1 male) who had symptomatic intracranial hypertension due to cerebral venous sinus thrombosis within a mean delay of 2 years. The mean age at onset was 31 years and the mean age on referral was 39.5 years. The predominant manifestation in our series were headache, papilledema, seizures and pyramidal syndromes. CT Scan showed non specific abnormalities in all of them and the sinus venous thrombosis was confirmed by MRI in 3 cases. The authors emphasize on the importance of MRI with angio MRI for the diagnosis, the outcome and the evaluation of the cerebral venous thrombosis after treatment. PMID- 12080562 TI - [Chronic pancreatitis in the ventral pancreas in pancreas divisum]. AB - Pancreas divisum, partial or non fusion of the dorsal and ventral pancreatic ductal system, affects up to 10 percent of the population. Approximately 25 percent of patients with pancreas divisum will develop complications such as recurrent pancreatitis. Surgical and endoscopic therapy usually include minor papilla sphincterotomy or sphincteroplasty to facilitate drainage of the dorsal ductal system. The following case represents an unusual complication of the pancreas divisum with primarily ventral duct disease and pancreaticolithiasis. PMID- 12080563 TI - [Catastrophic antiphospholipid syndrome. Report of two cases]. AB - The antiphospholopid syndrome has been associated with thromboembolic events, thrombocytopenia and fetal loss. Some patients with the antiphospholipid syndrome may develop an acutely catastrophic syndrome characterized by multiple vascular occlusions which often results in death. Most patients dye as a result of a combination of cardiac and pulmonary failure. Although trigger factors are present in a minority, in the majority the condition develops quite suddenly. Precipitating factors include infections and trauma (surgical). Treatment of the condition, once recognized, needs to be heroic. Plasmapheresis seems to be useful in several cases who had not responded to conventional therapy (heparin, steroids, immunosuppressive) directed against immunologically mediated intravascular thrombosis. We report here two cases of fatal catastrophic antiphospholipid syndrome and highlight the need for vigilance in the management of patients with antiphospholipid syndrome. PMID- 12080564 TI - [The association between Takayasu's arteritis and pregnancy]. AB - We present about three observations of patients suffering from TAKAYASU's arteritis two of home have necessitated one carotidien's angioplasties. From these three cases and from a review of literature, we analyse the different materno-foetal risks of the association TAKAYASU'S arteritis and pregnancy. The high arterial pressure and the pre-eclampsia constitute the aim maternal complication. The vascular accident and the cardiac decompensation can be seen in five percent (5%) of cases. The back wordiness of the intrauterine growth the major foetal complication. Eventually, we suggest the behaviour to hold in order to improve the materno-foetal prognostic. PMID- 12080565 TI - The nursing shortage: Part 2. Healthcare facilities take innovative approaches to recruitment and retention. PMID- 12080566 TI - Competency in the care of older patients. PMID- 12080567 TI - A matter of time. PMID- 12080568 TI - A new transcontinental disease: the West Nile virus. AB - The West Nile Virus caught New Yorkers by surprise in the summer of 1999. The prompt reporting of cases, as well as swift responses from local and state health departments and the Centers for Disease Control and Prevention, can prevent larger outbreaks of the virus. PMID- 12080569 TI - Pharmacogenomics: an evolving paradigm for drug therapy. AB - Individual variations in drug responsiveness are becoming better understood as a result of advances in genetics. The new science of pharmacogenomics capitalizes on that knowledge and promises safer and more powerful drug therapies. PMID- 12080570 TI - Gender and acute myocardial infarction symptoms. AB - Heart disease is the nation's most significant health problem. Unfortunately, the vast majority of studies conducted to explore the underlying the pathophysiology, diagnosis, and treatment of cardiac disorders are based on male samples. Exploring gender-based differences in symptom experiences will lead to improved research, clinical, and prevention initiatives. PMID- 12080571 TI - Rhabdomyolysis: the hidden killer. AB - Rhabdomyolysis can strike anyone given the right circumstances. Combining vigorous exercise, drugs, and extreme heat puts one at risk. However, there are many causes for "rhabdo." With early recognition and treatment, a devastating outcome can be prevented. Education is the key! PMID- 12080572 TI - Teaching staff a brief cognitive-behavioral intervention. AB - Lifestyle behaviors contribute significantly to the onset and course of numerous diseases and affect well-being in general. Patient education is the principal strategy nurses use to modify individuals' knowledge, attitudes, and beliefs about their health behaviors. However, current approaches are failing to induce the behavior changes necessary to reduce the incidence and prevalence of lifestyle-related disorders. In this pilot project, a brief cognitive-behavioral program was developed. The project yielded important insights into ways to improve current and develop new and more effective patient education strategies. PMID- 12080573 TI - [Agreement on the Barthel Index. A rapid analysis of other and self-assessment in elderly stroke patients]. AB - The relationship between staff and patient ratings of activity restrictions (Barthel Index, BI) was investigated in 120 elderly stroke patients (on average 78 years old) using the Rasch model and the rating scale analysis. In addition, the relationship between the rated activity restrictions and measures from the Geriatric Assessment was analyzed. We found good patient-staff agreement (r = 0.90) with the poorest agreement in the item "bathing" of the BI. There was also a highly significant correlation between staff and patient ratings and the Tinetti Gait and Balance Scales (r = 0.72 and r = 0.76, respectively). Correlations between other measures of the Geriatric Assessment and the rated activity restrictions were low explaining less than 8% of variance. Our findings merit the use of patient ratings of activity restrictions in stroke outcome research. However, self-ratings of activity restrictions were measured by an unstructured interview and it cannot be ruled out that this method had an influence on the correlation between self-rating and staff rating. PMID- 12080574 TI - [Relevance of neuropsychological studies of depression in the elderly]. AB - Neuropsychological deficits are a common feature of depressive disorders. In the systems ICD 10 and DSM IV neuropsychological deficits constitute a relevant diagnostic criterion. Deficits in executive functions, attention and memory are documented. Primarily, mental flexibility seems to be impaired. The relation between neuropsychological deficits and depression is moderated by the patient's age. In the elderly, depression is more often accompanied by neuropsychological deficits. Clinically, the relevance of neuropsychological testing in depression has different aspects. Neuropsychological testing is the prerequisite for neuropsychological therapy when patients exhibit residual cognitive symptoms, and for professional and social rehabilitation. The neuropsychological outcome provides important information for the therapy process, and for predicting therapy success. In addition, neuropsychological testing can influence the choice of medication and psychotherapy. Neuropsychological data can also help to differ depression from dementia, an important problem in elder psychiatric and neurological patients. For this purpose a comprehensive neuropsychological test battery that covers different neuropsychological domains needs to be employed. Furthermore, the analysis of the patients behavior and errors during testing provides essential information. However, in many cases a neuropsychological re test and the analysis of the course of neuropsychological deficits is needed to differ depression from dementia. PMID- 12080575 TI - [Concept and effectiveness of a treatment program for patients with dementia and their relatives. Results from the Bad Aibling Alzheimer Disease Therapy Center]. AB - A short-term residential treatment programme designed to prepare patients with dementia and caregivers for life with a progressive disease was evaluated in a pre-treatment post-treatment design. The multicomponent programme included: 1) intensive rehabilitation for patients, based on the concept of Self-Maintenance Therapy, and 2) an intervention programme for caregivers. Based on the concept of Self-Maintenance Therapy, it was hypothesised that experiences which are in accordance with patients' self-system would support patients' well-being, reduce psychopathological symptoms and facilitate social participation. In this study treatment effects in FTLD are analysed and compared with treatment effects in AD. The results for both groups show a consistent improvement in patients' depression and in other psychopathological symptoms, which can be seen as directly beneficial for patients. Following treatment, caregivers also felt less depressed, less mentally fatigued and restless, and more relaxed. It was predicted that FTLD patients challenge caregivers in a different way compared to patients with AD due to the particular profile of cognitive and psychiatric problems but there are no significant differences between groups. Further controlled follow-up studies are needed to evaluate the effects of the intervention as dementia progresses and to address hypotheses about factors responsible for benefits as well as for treatment resistance. PMID- 12080576 TI - [Intergenerational equity is more than pension financing]. AB - Public old age pensions--often termed the "generational contract"--are part of a broader system of exchange among generations which must be assessed in its entirety when issues of generational equity are at stake. The paper discusses the elements of this exchange system on the basis of the German Aging Survey, a large representative survey of the German population aged 40-85. Its empirical foci are two domains that are usually neglected in the public discourse on generational equity: the productive activities of the elderly and the transfers between generations in the family. The results demonstrate that the elderly contribute substantially more to societal welfare than has become visible so far. PMID- 12080577 TI - [Oxidative stress, age-dependent [correction of age-related] cell damage and antioxidative mechanisms]. AB - Cellular oxidative stress is due to the production of reactive oxygen species (ROS), on the one hand, and weaknesses of the antioxidative defence, on the other. This is particularly true for cells with an active metabolism such as neurons and muscle cells, but it is also relevant for all other cell types. Hydrogen peroxide is an important member of ROS and is generated predominantly by mitochondria. In combination with reduced trace metals such as iron or copper, hydrogen peroxide is transformed into the highly reactive hydroxyl radical which causes damage to virtually all macromolecules. Oxidation of nucleic acids results in mutations while protein denaturation leads to enzyme defects and impairment of the cytoskeleton. Lipid peroxidation in cell membranes is strongly involved in the perturbation of ion homeostasis. Because this cell damage ultimatively causes cell death, oxidative stress initiates several diseases. Mitochondria play a major role in this context because they are the main source of endogenous oxidative stress and additionally function as an inducer of programmed cell death (apoptosis). Several strategies of antioxidative defence exist: While transition metals can be inactivated by chelating proteins (e.g., ferritin), ROS can be reduced enzymatically (e.g., by the glutathione peroxidase) or non-enzymatically by antioxidants (e.g., by vitamin E, vitamin C and glutathione). Stress proteins are implicated in the repair and transport of denatured proteins as well as in the inhibition of apoptosis. PMID- 12080578 TI - [Effects of physical training and age-induced structural and functional changes in cardiovascular system and skeletal muscles]. AB - Age-related morphological changes in the cardiovascular system refer to a thickening of the arterial wall as well as a diffuse increase in the wall thickness of the myocardium. In terms of function, this leads to a decrease of the myocardial contractility in systole and to a delay of the diastolic myocardial relaxation. At the arterial wall, an impairment of wall elasticity as well as a decrease of the vasoreagibility can be observed. The systolic blood pressure and the blood pressure amplitude rise; the blood flow-dependent vasodilatation mediated by endothelial cells decreases. The loss of cardiac pacemaker cells by approximately 90% from the 2nd to the 8th decade leads to a reduction of the heart rate variability and to a decrease of the maximum heart rate during exhaustive physical exercise. Hemodynamically these changes resulted in a continuous decline of the maximum cardiac output as well as the maximum oxygen transport capacity. Between the 3rd and 8th decade the peripheral skeletal muscle mass decreases as a result of a reduction of the size and the number of muscle fibers with an overproportional loss of 30-40% regarding the highly glycolytic type IIb fibers. Furthermore, the intramuscular capillarization declines so that, with a concomitant decrease in the number and size of the mitochondria, the supply with oxygen and energy sources and consecutively the aerobic muscular energy metabolism decreases by 8-10% per decade beginning with the 4th to 5th decade. Additionally, the maximum and explosive muscular strength decreases by 30-35% from the 4th decade onwards. In older, healthy men who perform a regular aerobic muscular training in endurance sport disciplines (e.g., walking, jogging or cycling), a decrease of the resting heart rate, a restoration of the primarily lowered heart rate variability, an improvement of the early diastolic left-ventricular filling as well as a significant increase in the VO2max could be proven. Other studies showed that regular endurance training on a cycle-ergometer resulted in a significant increase in capillarization within the exercising muscle groups. In older, trained individuals a reduced stiffness of the arterial vessel walls and an improved maximum exercise-induced muscular blood supply occurs. In other training studies performed with older subjects, a positive influence of regular endurance training on the carbohydrate metabolism along with an improvement of the primarily reduced insulin sensitivity could be demonstrated. In older men and women who regularly underwent muscular strength training, a significant increase in maximum strength with a significant increase in the proportion of type II muscle fibers was found. PMID- 12080579 TI - [Effects of autogenic training in elderly patients]. AB - Autogenic training (AT) is a widely available relaxation method with beneficial outcome on physiological and psychological functioning. In our study, we wanted to test the effects of an AT course in cognitively impaired, frail elderly. After a 3 month waiting period (control), AT courses (intervention) of 3 months duration were offered in 2 nursing homes. Thirty-two frail elderly took part in the study, 24 of them had a psychiatric diagnosis (mean age 82.1 +/- 7.2 years, CAMCOG 75.5 +/- 15.7, MMSE 23.3 +/- 4.3, HAMD 10.0 +/- 3.6, NOSGER 57.2 +/- 18.4, AT-SYM 32.9 +/- 17.6 points). Eight participants dropped out during the waiting period, 8 during the course. From the 16 participants, 15 (94%) were able to learn the AT according to subjective, 9 (54%) according to objective criteria. The ability to practice the AT successfully correlated with the CAMCOG (p = 0.001) and the NOSGER (p = 0.01) score. Participants with a dementia syndrome had major difficulties, whereas age, depressiveness, and number of complaints (AT SYM) had no influence on the ability to learn the AT. There was no intervention effect, measured with the HAMD, NOSGER, AT-SYM and MMSE. In the pre-post comparison of training sessions, a significant improvement in general well being was found (p < 0.001). Mentally impaired, frail elderly participants are able to learn the AT. Cognitive impairment is disadvantageous for a successful participation. PMID- 12080580 TI - [Number processing and acalculia in dementia]. AB - In this review, studies on number processing and calculation in demented patients are summarized. Single cases, group studies and our own examinations all show that acalculia is a frequent symptom of dementia and correlates with severity of dementia. While calculation and numeral transcoding deficits are often prominent in early courses of the disease, deficits in semantic processing and basic number processing are less severe. Still, single cases demonstrate that very specific dysfunctions in number processing and calculation can occur in dementia. In number transcoding, a characteristic error type occurs in demented patients that has not (or rarely) been described in other brain damaged people so far. In these "shift errors" or "intrusion errors" the wrong number code is used or the number codes are mixed (e.g. patients write "2hindert11" instead of zweihundertelf (two hundred eleven)). Due to the frequent occurence of acalculia in dementia and its specific features, it is suggested that acalculia should be integrated into the criteria for dementia and that number processing and calculation tests are used in the diagnosis of dementia. PMID- 12080581 TI - A great achievement: Latin American Journal of Nursing enters the SciELO collection. PMID- 12080582 TI - U.S. nursing students in Nicaragua: a community health clinical experience. PMID- 12080583 TI - [Trends and perspectives of nursing administration: a study in the Santa Casa hospital of Belo Horizonte-MG]. AB - The "Santa Casa" Hospital of Belo Horizonte has implemented a new management system that has characterized it as a flexible organization. Based on this context, the authors of this research defined the following objectives: to analyze nursing administration by considering the implementation of the Management System (SIGESC), to describe how the nursing team perceives communication, the decision-making process and interpersonal relationships, to identify possible trends and perspectives present in the administrative practice developed. A time of transition in nursing administration that presents characteristics of classical management and contemporaneous management was shown as well as trends and perspectives that will contribute to the reorganization of nursing work in hospitals. PMID- 12080584 TI - [Epidemiological surveillance and the perspective of field working -- Riberao Preto Municipal Health Secretariat]. AB - This study aimed at analyzing the conformation of the Epidemiological Surveillance Service in Ribeirao Preto Municipal Health Secretariat under the orientation of the principles established by the Unified Health System and the perspective of Health Surveillance in face of the health care municipalization process occurring in the 1990s through the following dimensions: territory; problems; intersectoriality based on positive health concepts and on the paradigm of social health production. By means of a qualitative approach, bibliographical and documentation sources as well as 17 semi-structured interviews were used. Data were analyzed through the thematic analysis technique by which the following themes were identified: from "Monitoring" to "Educating and Preventing": practice in Epidemiological Surveillance--a proposal to work in the territory. It was concluded that the potentiality of actions related to Epidemiological Surveillance is associated with the perspective of working in the territory for the construction of Health Surveillance in order to assure the technical content of those actions as well as the participation of different professional categories in the same work project, which will not necessarily be performed by all professionals in the same health production process, territory or time. PMID- 12080585 TI - [The use of home inquiry as a tool to follow up health actions in microareas -- analyzing the vaccination status of children under 1 year of age]. AB - One of the strategies on which local health services rely in order to follow up activities is home inquiry, a source of important information concerning the access to services and knowledge regarding the population's health conditions. This work aims at analyzing the use of inquiries as an instrument for diagnosis and follow-up of the vaccinal situation of children under one year old in a micro area of the City of Ribeirao Preto-SP, Brazil. In the first semester of 1998, 106 families were interviewed by using a family record. Nine children under 01 year old (4.35% of the population) were identified, of whom 4 (44.44%) presented vaccination delay. It was later identified that 8 children presented an updated record and that information was not obtained in only one case. It can be verified that home inquiry is a viable strategy to follow up and evaluate the care delivered, which, in the case of this study, concerns vaccination actions, so as to efficiently and effectively determine the directions to be taken in health care in local level. PMID- 12080586 TI - [The meaning of ostomy care among nurses: a case study]. AB - This study aimed to identify how nurses working in a hospital interpret ostomy care. The case-study methodology was used. The obtained results originated four categories: the patient needs special care, it is necessary to provide orientation, in order to provide orientation, the nurse needs..., the recognition of difficulties in delivering special care. These categories express that the care given to ostomy patient by the nurses is not appropriate due to the lack of specific knowledge. PMID- 12080587 TI - [Study of accidents involving children assisted at an emergency ward]. AB - The aim of this study was identify cases of accidents victimizing children assisted at the emergence ward of a general hospital. Data were collected from January to March, 1999 through a protocol with variables related to the identification of the accidents as well as of the children. The sample in the study consisted of 890 children of whom 56.1% were males and 43.9% were females. The most prevalent accident was fall (46.9%), the age with a major incidence was between 7 and 11 years old, the parents were present to 43.4% of the cases. The children were discharged from hospital after being given care in 95.7% of the cases. PMID- 12080588 TI - [Determinant beliefs nurses' intentions concerning the presence of parents in neonatal intensive care units]. AB - This study aims at learning about the beliefs, attitudes and social rules which determine the nurse's intention to allow the presence of parents in Neonatal Intensive Care Units. The content of semi-structured interviews conducted with 11 nurses was analyzed by using the Rationalized Action Theory as a theoretical framework. The thematic units enabled the comprehension of the relationship between the nurse's intention and behavior when allowing the presence of parents in Neonatal Intensive Care Units as well as the influence of the nurse's personal beliefs and subjective norms on her behavior. PMID- 12080589 TI - [Brazilian scientific production on nosocomial infections and nursing contribution: past, present, and perspectives]. AB - This article analyzes the Brazilian scientific production concerning nosocomial infection from 1950 to the present day and emphasizes the contribution from Nursing. The theoretical and analytical resource used was its contextualization with the country's social development and health policies, which enabled to demonstrate the close relationship between the development of nosocomial infection, the scientific production on such theme and the health care models adopted. The importance and limits of this scientific production are discussed as well as the need to include nosocomial infection in a larger field of care quality in general. The article also verified that the latest data on Nursing already show this tendency. PMID- 12080590 TI - [Occupational trauma of the vertebral column in nursing workers]. AB - All occupational accidents (CAT) reported at a University hospital, from January 1990 to December 1997, were analyzed and the characteristics of the vertebral column trauma caused by the occupational accidents involving members of the nursing team were investigated. During this period, 531 accidents were reported and 37 (7%) of these were typical vertebral column traumas. These results suggested that the number of accidents reported were below actual estimates and that the nursing auxiliaries were the most affected. The accidents were mainly caused by falls and during the transport or transfer of patients and equipments. PMID- 12080591 TI - [Psychiatric nursing consultation and liaison: how do nurses understand this activity?]. AB - This research aimed at identifying the knowledge of nurses working in medical and surgical clinics and with burned patients as to psychiatric nursing consultation by means of a semi-structured interview. Care situations considered to be difficult were related to instrumental resources and provision of psychological support. Most of the nurses try to listen to patients' complaints, but they do not know what else they could do. Whenever they need help, they think of colleagues, bosses, psychologists and psychiatrists. They know the specialty of the psychiatric nurse and believe that this professional can help with emotional support to the patient and the nursing team. They consider consultation and liaison to be an evaluation of the patient by another specialist. PMID- 12080592 TI - [Sexuality of individuals with the human immunodeficiency virus (HIV): study based on the crisis theory]. AB - The purpose of the investigation was to identify and analyze the sexual difficulties and their relation to the social and emotional difficulties experienced by subjects during a HIV crisis. Data were collected through semistructured interviews taped, transcribed, categorized and analyzed. Authors transcribed the conversations using thematic analysis and created five stories entitled: ... "Living with HIV". Data analysis was based on the interpretation of the content associated to the crisis theory. Data enabled authors to conclude that subjects undergoing a HIV crisis did not achieve a positive level of sexual, social or emotional adaptation. The speech of these subjects clearly showed the time of HIV crisis characterized by mechanisms of coping, usually negative. The results also indicate the need to rethink nurse's role, specially nursing intervention. Authors believe that the Model of Mental Health Nursing Procedures is adequate to help subjects in a HIV crisis to solve their sexual, emotional and social difficulties and to search for a positive level of adaptation, considering the difficulties that are inherent to the disease. PMID- 12080594 TI - [Ethical aspects concerning nurses' managerial activity]. AB - Aiming at addressing the ethical aspects concerning nurses' managerial activities, the authors of this work referred to Etzioni and adopted a hospital classification as a reference of organization in which consent is based on the internalization of rules accepted as legitimate. Regarding patients, their need to adapt to medical behavior and hospital rules through internalization is considered to be a result of physicians', nurses' and the hospital's power. However, the authors view that such internalization is naive, without consent, especially by taking into account that most Brazilian patients do not know that they should or should not consent and are used to obeying. Thus, the work focused on nurses' managerial actions, indicating that they must be based on professional values, the Nursing ethical code as well as on the rights of hospitalized patients, thus integrating qualified care guided by respect, free consent and promotion of patients as the protagonists and subjects of care. PMID- 12080593 TI - [Analysis of titles of research articles published in a Brazilian nursing journal]. AB - This study analyzes titles of research works published in a nursing journal in 1997 and 1998. Data were collected through forms and organized by using the EPI INFO 6.0 computer program. The analysis was based on recent national and international literature concerning the analysis of research reports. Results, which have been disposed in tables, showed that most of the analyzed articles presented a partly adequate titles in relation to variables, studied populations and/or key words. It was concluded that the evaluation of titles in nursing research still requires more attention since a large number of databases used by researchers only publicize research work means of its titles. PMID- 12080595 TI - [Analysis of the negotiation/accommodation concepts of M. Leininger's theory]. AB - This work aims at clarifying the method of implementation of the negotiation/accommodation care preconized by Madeleine Leininger's theory concerning the Universality and Diversity of Care. The conceptual analysis technique is used as a tool to analyze the literature. It was verified that the concept of accommodation defines the idea of adaptation, of something passive, whilst in negotiation, each party commands resources and needs to be fulfilled. Among the different works which were analyzed and used Leininger's theory, only one showed to approach negotiation. As a conclusion, accommodation is considered to be different from negotiation, the former being more commonly seen in practice than the latter. PMID- 12080596 TI - [Conceptual model of interactive open system and the theory of goal attainment of Imogene King]. AB - This work describes the Conceptual Model of Interactive Open Systems and the Theory of Goal Attainment by Imogene King by reflecting on the care process based on this proposal. It was accomplished as a result of the extensive study of King's theoretical structure and its application with patients. It was possible to observe that the theoretical structure supplies the necessary theoretical and conceptual framework which is necessary for humanized nursing care. Also, it remains updated and included in the current context of health care. PMID- 12080597 TI - Peer review in scientific dissemination. PMID- 12080598 TI - [Development and evaluation of an educational software for pediatric nursing education]. AB - Computer Science is being introduced in education at an accelerated pace, thus forcing educators and students to become familiarized with such technology. This study aimed at the development and evaluation of the educational software "Children and medication". The program utilizes multimedia resources in four topics: the child, medication, the person who administers and administration of medication. The evaluation showed a large acceptance of the program, with most of the items being rated as excellent. We emphasize the importance of the development of teaching strategies utilizing computer resources in nursing education and in the development of nurses for the new millenium. PMID- 12080599 TI - [Nurses' professional prestige: estimate of magnitudes and expanded categories]. AB - The prestige of professionals such as social workers, biologists, dentists, nurses, engineers, pharmacists, physicists, physical therapists, speech-language pathologists, physicians, psychologists, chemists and sociologists was scaled by the psychophysical methods of estimation of magnitudes and expanded categories. Results showed that: 1) when we increase the limited amplitude of categories, this method has the same characteristics as those of the estimation of magnitudes. 2) the relationship between the estimations of magnitudes and estimations of expanded categories is a power function with an exponent that is not significantly different from 1.0. These data enabled the following conclusions: 1--The nursing profession is in the seventh or eighth position regarding the prestige of the 13 professions whereas physicians are in the first position in the scale obtained by the used methods; 2--the orders resulting from the methods produce positions of prestige that highly agree for the different professions. PMID- 12080600 TI - [Analysis of difficulties related with the daily activities of adolescents and children with type-1 diabetes mellitus: mothers' accounts]. AB - This descriptive study aimed at analyzing the difficulties faced by mothers of type-1 diabetic teenagers and children in view of the child's daily activities and based on the principle that emotional, social, personal and economic factors concerning the family interfere with achieving good metabolic control. Data were obtained by means of a semi-structured interview guide applied to 30 mothers. Data analysis followed the standards preconized for descriptive studies in which directed interviews are used. Results showed that: mothers have difficulties in dealing with children's daily activities such as social gatherings, in motivating them to develop physical activities, diet, school adjustment, relationship with the health team as well as with the children's behaviors regarding their brothers/sisters. Results indicated the need for planned and integrated work by a multiprofessional team with the families, considering all the aspects addressed in this study and their interdependence, aiming at a successful care to diabetic children and adolescents. PMID- 12080601 TI - [Problems during the recovery of patients following coronary artery bypass surgery: the follow up by nurses in the first month after hospital discharge]. AB - Patients may present problems after coronary artery bypass surgery and nurses play an important role in the detection of such problems. With the purpose to determine the problems of 25 patients during the first month after hospital discharge, they were followed up via telephone as well as when they returned to hospital. Content analyses of detailed nurses' recording revealed eight categories of problems. The most frequently related problems were related to the surgical incision site (76%), pharmacological therapy (28%), other pathologies (24%), mood alterations (20%) and sleep pattern disturbances (16%). The strategy chosen showed to be efficient to analyze the recovery of these patients during the first month after hospital discharge. PMID- 12080602 TI - [Self-esteem in adolescents with and without cleft-lip and/or palate]. AB - Self-image is an important determinant of self-esteem among men and women, which makes us reflect about the process of rehabilitation of people with facial malformations. In order to measure self-esteem, the Janis and Field Scale was used. The sample consisted of 608 adolescents aged 17 to 20 years including males and females. Two hundred and thirty-five had a cleft lip and/or palate and 373 did not. The analysis of the self-esteem levels obtained enabled to conclude that the adolescents with a cleft lip and/or palate behave differently from those without it, that is, they present lower self-esteem scores. PMID- 12080603 TI - [Analysis of attitudes of nurses and nursing students concerning computer use in Paraiba-BR]. AB - In this study an attempt was made to analyze the attitudes of both nurses and Nursing students in relation to computer use. Demographic data and attitudinal scaling were used. Three factors were found for nurses: obstacles to computer use; the computer as a work tool; the effectiveness of information to Nursing. As to Nursing students, in addition to these three factors, another one was also found: the need for educational training in computer technology applied to Nursing. Nurses' and students' attitudes were similar. It was, therefore, concluded that both groups have moderate attitudes concerning computer use, although the students showed more familiarity. PMID- 12080604 TI - [Domiciliary assistance in natural delivery]. AB - This study presents the experience of an obstetric nurse and of a couple who had their third child. It focuses on care standards that value women and couples in the childbirth and delivery process. It describes the care given to a couple during the pregnancy stage and during home delivery. The experience enabled the active participation of the couple and their children in the childbirth process, which essentially provided satisfaction to the family and to the professional. PMID- 12080605 TI - [Tetanus in the geriatric population: is it a collective health problem?]. AB - Tetanus is an infectious non-contagious disease caused by the bacillus Clostridium tetani, which penetrates in the organism through wounds. Psychomotor dysfunction facilitates accidents among elderly people and vaccinal coverage is low in this population, thus contributing to high lethality. This study aimed at reflecting on the situation faced by elderly people in relation to tetanus in the perspective of Collective Health. It is a Case Study conducted with two elderly males who had accidental tetanus and were hospitalized in a hospital in the municipality of Fortaleza. Data collection took place from March to April, 1998. The analysis showed the absence of vaccinal coverage as well as of the implementation of emergency prophylaxis. The two patients' conditions developed to death, which confirmed the high mortality due to tetanus in this age group. The critical reflection pointed out the urgency of a collective health approach. PMID- 12080606 TI - [Nursing understood as a vocation and its relationship to nurses' attitudes towards work conditions]. AB - This study aimed at reflecting about nursing as job by disconsidering the widely accepted conception that identifies it as an assistance activity for that conception might hinder actions towards coping with and changing work conditions. The study attempted to place in history the context in which the vocational/religious model was elaborated as well as to identify the manifestations of that conception in the discourse of Nursing freshmen and seniors, pointing out the fact that the teaching activity must be permeated by the conception of nursing/work. PMID- 12080607 TI - [Neurological and neuropathological studies of amyotrophic lateral sclerosis/parkinsonism-dementia complex in the Kii Peninsula of Japan]. AB - Hohara and Kozagawa in the Kii peninsula of Japan are reported to be high incidence foci of amyotrophic lateral sclerosis (Kii ALS) and parkinsonism dementia complex (Kii PDC). During the period between 1996 and 1999, three Kii ALS patients and 19 Kii PDC patients were confirmed neurologically in Hohara among which, one Kii ALS patient and two Kii PDC patients were examined neuropathologically. The ratio of positive family history where ALS or PDC occurred within the fourth degree of the relatives was 33.3% in the patients with Kii ALS, 78.9% in those with Kii PDC, and 72.7% in total. The ages of onset were between 57 years and 63 years (mean age: 60.0 years) in the patients with Kii ALS and between 53 years and 74 years (mean age: 66.5 years) in those with Kii PDC. All of the Kii ALS patients were female, and the male to female ratio of the Kii PDC patients was 1:1.7. The clinical features of Kii ALS were basically similar to those of classical ALS. The core clinical features of Kii PDC consisted of dementia and parkinsonism, frequently accompanied by motor neuron symptoms. The cardinal neuropathological features of Kii ALS/PDC included many neurofibrillary tangles (NFTs) associated with loss of nerve cells in the cerebral cortex and the brain stem, as well as morphological alterations diagnostic of ALS. Ultrastructurally, NFTs consisted of paired helical filaments. When we compared the clinical features of these Kii ALS patients with those that had been surveyed in 1969, the male to female ratio changed from male dominance to female dominance and the mean age of the onset of the disease was delayed by approximately 10 years. The most frequent initial symptom had been weakness of the lower limbs in the survey in 1969 and was bulbar palsy in this study. As to Kii PDC, this is the first report of the clinical features of many cases. PMID- 12080608 TI - [A case of spontaneous intracranial hypotension with simultaneous CSF leaks at the upper and lower thoracic levels]. AB - We presented a patient of spontaneous intracranial hypotension (SIH), in which radionuclide cisternography demonstrated multiple CSF leaks. A 51-year-old previously healthy woman awoke with a severe headache and nausea that were almost completely relieved by lying flat (day 1). The headache was not relieved by analgesics. On day 10, she visited our clinic. Neurological examination showed no abnormalities. A lumbar puncture revealed an opening pressure of 50 mmH2O, protein level of 64 mg/dl, 21 erythrocytes 21/microliter, and 4 lymphocytes/microliter. Cranial MRI on day 11 with gadolinium infusion demonstrated diffuse thickening and enhancement of the meninges. No signs of venous sinus thrombosis were identified. There was no downward displacement of the cerebellar tonsils. Indium-111 radionuclide cisternography demonstrated early accumulation of the tracer in the urinary bladder and CSF leakage at the upper thoracic level on the left side, at the lower thoracic level on both sides, and at the lumbar level. It is noteworthy that the scintigram taken 4 hours after tracer injection demonstrated CSF leakage at the injection site but not in the upper thoracic area. The patient kept sitting for about 20 minutes just before the images were obtained. Spinal MRI did not show meningeal diverticulum and epidural fluid collection. Ten days of bed-rest and transfusion failed to relieve the headache. Epidural patching with 7 ml of autologous blood performed at the Th2/3 interspace partially relieved the headache. Five days later, a second epidural patching was performed at Th11/12, which was slightly effective. A third epidural patching at L3/4 was ineffective. On day 32, a fourth epidural patching was performed at Th1/2 while the patient was lying down on her left side. The postural headache disappeared. Repeated cranial MRI with gadolinium infusion performed on day 47 showed resolution of the abnormal findings. To our knowledge, this is the second case report of SIH with double CSF leaks. To localize multiple leaks, radionuclide cisternography should be performed with a patient in various positions just before images are obtained. PMID- 12080610 TI - [A patient with intravascular malignant lymphomatosis presenting subacute dementia one year after sustained urinary retention and high serum LDH activity from the onset]. AB - We report a 71-year-old man with intravascular malignant lymphomatosis who showed high serum LDH and urinary disturbance for one year before manifesting dementia. High serum LDH was found at a health check at age 70. Two months later, he had an onset of backache and urinary retention. MRI of the spinal cord was unremarkable. One year later, he showed decline of mental activities and was admitted to our hospital. He was agitated and confused. However cranial nerve palsy or limb weakness was not noted. The MRI of the brain showed T2-high signal in bilateral occipital, right temporal lobe and the left insular cortices. The abdominal CT scan showed swelling of the adrenals on both sides. Adrenal biopsy revealed diffuse large B cell lymphoma. He developed respiratory distress and he died two months after the admission. Post mortem examination revealed intravascular and extravascular proliferation of lymphoma cells in most of the internal organs including adrenals, spleen, liver and the kidneys. In the brain, the laminar necrosis was seen in the left occipital cortex and hemorrhagic infarctions were noted in the insular and temporal cortices and the medial temporal cortex. Sacral spinal cord showed necrosis of the gray matters and loss of myelinated fibers in the white matter. Intravascular proliferation of the lymphoma cells were also seen in the vessels of the brain and the spinal cord. This patient suggests the importance of survey for intravascular malignant lymphomatosis, when high serum LDH and myelopathy of lumbosacral area are seen. PMID- 12080611 TI - [A case of acute disseminated encephalomyelitis after renal transplantation]. AB - We report a patient of acute disseminated encephalomyelitis (ADEM) in a recipient of renal transplantation. A 43-year-old man suffered from generalized convulsion and consciousness disturbance followed by coma on day 53 of after the transplantation. He was receiving several immunosuppressants, and an increase of serum antigen for cytomegalovirus was observed one day before the ictus. T2 and diffusion-weighted image of MRI showed high intensity lesions in the bilateral cerebral white matter, basal ganglia, thalamus, midbrain, pons and cerebellum. Examination of cerebrospinal fluid revealed elevated myelin basic protein level. The patient was diagnosed as having ADEM and was treated with methylpredonisolone pulse therapy in combination with intravenous immune globulin. He gradually recovered and became capable to eat and sit on a wheel chair. White matter lesions on MRI were also diminished. ADEM may occur in recipients of organ transplantation even if they have immunosuppressive treatment. PMID- 12080609 TI - [A case of late-onset carbamoyl phosphate synthetase I deficiency, presenting periodic psychotic episodes coinciding with menstrual periods]. AB - Carbamoyl phosphate synthetase I deficiency (CPSID) is a rare metabolic disorder affecting the first enzymatic step of urea cycle. We report clinical manifestations of a female case of late-onset CPSID in Japan. An 18-year-old girl was admitted to emergency room due to acute comatose state. Her parents had no apparent consanguineous history. She had suffered from intermittent psychotic episodes (excitation, aggressive behavior and insomnia) with nausea and vomiting from the age of 13, mostly coinciding with menstrual period. She had minor learning disability without major neurological deficits and convulsions. Her mental status was estimated as normal in her intermenstrual period. She had been diagnosed as having hysteria and premenstrual syndrome. Her neurological findings on admission showed deep coma and hypotonic tetraparesis. Plasma ammonia level was markedly elevated (684 micrograms/dl) without significant liver dysfunction. Blood urea nitrogen decreased to 6 mg/dl. Continuous venovenous filtration with subsequential administration of sodium benzoate and l-arginine was started to eliminate blood ammonia. Although the plasma ammonia level decreased to 300 mu/dl in next 10 hours, severe cerebral edema was observed in head computed tomography subsequently, followed by marked cerebral atrophy. Finally, her consciousness status became almost alert a month after the onset, but her mental status was severely retarded. CPSI activity of her biopsied liver markedly decreased and she was diagnosed as having CPS ID. CPSI cDNA analysis of her biopsied liver demonstrated a V1149G mutation. Genomic DNA analysis showed that she was heterozygous in V1149G mutation. The mutation allele was derived from her father. The causative factor for absence or very low level of maternal CPSI mRNA will require further analysis. PMID- 12080612 TI - [A case of recurrent aseptic meningitis caused by high-dose intravenous gammaglobulin therapy for chronic inflammatory demyelinating polyneuropathy]. AB - We experienced a patient of CIDP who was twice complicated with aseptic meningitis following high-dose intravenous gammaglobulin therapy. The patient was a 29-year-old woman who first developed gait disturbance in September 1998. Neurological examination revealed muscle weakness and sensory disturbance in the distal parts of four extremities and decrease of deep tendon reflexes. Cerebrospinal fluid analysis revealed an elevated protein content and a normal cell count. Steroid therapy was effective in early stage, however, this effectiveness had been reduced gradually. She received high-dose intravenous gammaglobulin administrations in September 1999. On the fourth day after start of this therapy, she developed severe headache, nausea and nuchal rigidity without fever. Cerebrospinal fluid analysis revealed an increased cell count of mononuclear predominance and an elevated protein content. As bacterial culture remained negative and viral titers were not elevated, aseptic meningitis was diagnosed. The therapy was stopped, and thereafter her headache continued for 7 days. The muscle weakness and sensory disturbance were remarkably improved, but 9 months later, her symptoms became worse again. She received high-dose gammaglobulin administrations for 2 days in June 2000 and developed aseptic meningitis again. Over again, 9 months later, she received the same medication for only 1 day in March 2001 and she developed mild headache but not meningitis. Aseptic meningitis with CIDP following high-dose gammaglobulin therapy was rare, however, we should pay attention to this therapy in patients with CIDP and may prevent the occurrence of aseptic meningitis by reducing the total dose and shortening the administration periods of gammaglobulin. PMID- 12080613 TI - [Anti-GQ1b IgG-negative case of overlapping Fisher's and Gullain-Barre syndromes after Campylobacter jejuni (PEN 19) enteritis]. AB - We described a 70-year-old woman with overlapping Fisher's syndrome (FS) and Guillain-Barre syndrome (GBS), from whom Campylobacter jejuni had been isolated. In typical FS as well as GBS with ophthalmoplegia and acute ophthalmoparesis without ataxia, serum anti-GQ1b IgG antibody often is detected and ophthalmoplegia is characterized by the predominant abducens palsy. This patient, however, showed marked oculomotor nerve disturbance. Serum anti-GQ1b IgG antibody was negative and IgG antibodies against GM1, GM1b, and GD1a were strongly positive. Although FS and overlap of FS/GBS have been reported to be associated with PEN2 of C. jejuni, the isolate from our case belonged to PEN 19. C. jejuni serotype may be associated with clinical manifestations and anti-ganglioside antibody species. PMID- 12080614 TI - [Successful treatment of emotional disturbances following non-herpetic limbic encephalitis by serotonergic agents. A case report]. AB - We report a 19-year-old woman who developed refractory status epilepticus due to non-herpetic limbic encephalitis. Because ordinary anti-epileptics were ineffective, general anesthesia under mechanical ventilation was begun with pentobarbital, midazolam, and propofol. After 4 months, we could finally discontinue the intravenous anesthetics. Then, she gradually became conscious and several weeks later, could communicate verbally. Simultaneously, she began to manifest psychomotor agitation and emotional incontinence mimicking Kluver-Bucy syndrome. Brain MRI revealed cortical atrophy in the fronto-temporal lobes and dilated anterior and inferior horns of the lateral ventricles. Dopamine blockers and benzodiazepines failed to resolve these emotional symptoms. Oral tandospirone (30 mg/day dose) was initiated and a partial regression was observed. The following administration of fluvoxamine (started with 12.5 mg/day and maintained with 75 mg/day) resulted in a dramatic improvement of her symptoms within 3 days. This clinical course suggests that impaired serotonergic neurotransmission plays a key role in her emotional disturbances and that its modulation by serotonergic agents is useful to relieve such symptoms. PMID- 12080615 TI - [A case with left mesial temporal lobe epilepsy characterized by abnormal massive salivation]. AB - A 26-year-old woman was admitted because of a 12-year history of intractable epileptic seizures. Every seizure began with an upper abdominal discomfort, and often followed by massive salivation. Whenever the epileptic salivation happened, the patient lost consciousness, and sometimes she developed generalized convulsions. In terictal sphenoidal EEG recordings revealed abnormal discharges in the left mesial temporal region. MRI demonstrated left hippocampal atrophy. Since her seizures were medically intractable, left temporal lobectomy was performed. Neuropathologic examination revealed hippocampal sclerosis. After the operation, she became completely seizure-free with no episode of massive salivation. From the literature, epileptic salivation can originates from the rolandic area; fronto-orbital cortex & cingulate gyrus; insula & operculum; and mesial temporal structures. The abnormal massive salivation in our patient might be attributable to the activation of frontal limbic system triggered by hippocampal abnormal firing during the ictal period of TLE. PMID- 12080616 TI - [Giant-cell myocarditis without a symptom of heart failure seen in a patient with myasthenia gravis and concurrent Hashimoto's disease]. AB - A 69-year-old Japanese woman initially noticed difficulty in squatting in the last two years, followed by nasal voice, fatiguability in mastication, and blepharoptosis. On admission to our hospital, in addition to these neurological findings, we detected ectopic arrhythmia and Levine II systolic murmur at the apex region, without any subjective symptoms. The serum titer of anti acetylcholine receptor antibody was elevated to 28 nmol/l (normal: < 0.2), and she responded to repetitive nerve stimulation at a frequency of 3 Hz showing 13% waning, she had positive test for edrophonium administration, and was diagnosed as having myasthenia gravis (MG). There was no thymoma by radiographic examination. She also had Hashimoto's disease confirmed by the laboratory findings. Chest X-p revealed dilatation of the heart, and 24-hour Holter ECG revealed non-sustained ventricular tachcardia (VT). Ventriculography revealed prominent dilatation of the left ventricle and diffuse hypokinetic ventricular wall motility. Endomyocardial biopsy revealed muscle fiber degeneration, cellular infiltration, and scattered multinucleated giant cells, confirming a diagnosis of giant cell myocarditis (GCM). Quadriceps muscle biopsy revealed a small number of muscle fibers with giant nuclei, but no giant cells were seen. Immunological study revealed elevation of CD4/CD8 ratio and memory CD4 cells. Antibody to anti cardiac and anti-striate muscle were strongly positive in the serum. Four months later, she developed dyspnea on effort and hypoxia, accompanied by severe bradycardia leading to sinus arrest. For acute cardiac deterioration, steroid pulse therapy was started followed by oral predonisolone and azathioprine, which aggravated myasthenic symptoms. The patient was ventilated for respiratory hypercapnia. During immunoabsobent therapy, she developed VT which caused a cardiac arrest, leading to fatal outcome. In case of MG, especially overlapped with other autoimmune diseases, evaluation of cardiac function should be conducted to detect GCM. PMID- 12080618 TI - [Sequential magnetic resonance imaging and neurophysiological studies in a patient with inferior spinal cord infarction]. AB - A 66-year-old woman suddenly developed anterior spinal artery syndrome with complete flaccid paraplegia, superficial sensory disturbance caudally to the L5 dermatome level with preservation of deep sensation, incontinence, and absent deep tendon reflexes in both legs. An MRI of the whole spine and an analysis of the CSF 4 hours after onset were normal. The electrophysiological study showed an absence of F wave on the posterior tibial nerve stimulation on admission, while the peripheral nerve conduction velocities and amplitudes of upper and lower limbs were normal. T2-weighted MRI 4 days after onset demonstrated an area of high signal intensity in the gray and white matters of the epiconus and conus medullaris, and T1-weighted MRI showed the swelling. Three weeks later, F wave became evoked nd the high signal areas on axial T2-weighted MRI were localized in the bilateral anterior horns of the gray matter. T1-weighted MRIs after an administration of Gd-DTPA 3 and 7 weeks after onset demonstrated an enhancement of the ventral roots of the lumbar nerves and cauda equina, while the enhancements disappeared 8 months after onset. The patient was finally able to walk independently over 10 meter. An absent F wave was an only positive finding at the hyperacute (hours after onset) stage of the spinal cord infarction. PMID- 12080617 TI - [A case of myasthenia gravis accompanied by large thymoma and anti-GAD antibody]. AB - A 61-year-old woman had repeated episodes of muscle weakness of face, neck and limbs for 18 years. She was diagnosed as having myasthenia gravis (MG) by the positive anti-acetylcholine receptor antibody and findings of electromyogram. Simultaneously, she was noticed to have diabetes mellitus with high titers of anti-glutamic acid decarboxylase (GAD) antibody. Magnetic resonance imaging showed a large thymoma. In spite of the improvement of MG after thymectomy, the insulin secretion slowly exacerbated during next two years. The clinical course of her disease was characteristic as slowly progressive insulin dependent diabetes mellitus (SPIDDM). She continued to have positive autoantibody against beta-cell of pancreas. Recently, anti-GAD antibody is detected in patients with SPIDDM and stiffman syndrome (SS) in high rate, and it is closely associated with the cause of these syndromes. The patient did not reveal the symptoms of SS. From the clinical course, MG and SPIDDM in this patient may be caused by a common underlying autoimmune abnormality resulting from the long presence of the thymoma. MG and SPIDDM may be derived from organ-specific autoimmunopathy from the defect of self-tolerance. PMID- 12080619 TI - [Laryngeal carcinoma--a question of personal susceptibility]. PMID- 12080620 TI - [Approaches to organ preserving therapy of carcinomas of the larynx and hypopharynx]. PMID- 12080621 TI - [Hereditary hemorrhagic telangiectasia (Osler's disease). An interdisciplinary challenge]. AB - BACKGROUND: Hereditary hemorrhagic telangiectasia (HHT or Rendu-Osler-Weber disease) is an autosomal dominant inherited disease of the fibrovascular tissue. More than 90% of patients have recurrent nosebleeds. Therefore, ENT physicians often have a key position for diagnosis and management of the disease. Epistaxis can severely reduce the quality of life, but visceral lesions are also life threatening. METHODS: This paper provides a review of the literature about the disease, its diagnosis, preventive measures, and therapy of the different manifestations. RESULTS AND CONCLUSIONS: Physicians should be especially aware of gastrointestinal telangiectases, arteriovenous malformations (AVM) of the lungs, liver, and brain. A screening is recommended at least for pulmonary AVM. PMID- 12080622 TI - [Measuring beta-trace protein for detection of perilymph fistulas]. AB - BACKGROUND: beta-trace protein is an immunological marker for cerebrospinal fluid or inner ear fluids. We present two initial cases in which a beta-trace protein nephelometric assay provided immunological proof of perilymph in samples taken from the nasopharynx. PATIENTS AND METHODS: Two cases are presented of an acute vestibulocochlear disorder following stapedotomy. Fluid samples, taken from the nasopharynx, were investigated using beta-trace protein laser nephelometry. The samples were collected using Merocel tamponades, which were left in place over night. RESULTS: In both cases, the samples were positive for beta-trace protein. Tympanoscopy was performed and the oval window was closed. After tympanoscopy a partial recovery of the hearing impairment was observed. CONCLUSION: Using beta trace protein, it seems to be possible to diagnose a perilymphatic fistula by noninvasive means. PMID- 12080623 TI - [Tracheal ruptures in endotracheal intubation. Diagnosis and therapy]. AB - BACKGROUND: Tracheal ruptures are most frequently the result of a blunt trauma to the thorax or of forced intubation. They represent a rare, but life-threatening complication that requires immediate help. METHODS: The resulting pneumothorax and the skin/mediastinal emphysema are the most prominent clinical signs. Injuries of the tracheobronchial tract should be diagnosed endoscopically and treated surgically immediately after the trauma. RESULTS: The present paper reports on three cases of tracheal ruptures after forced intubation which could be successfully managed by tracheostomy and subsequent reconstruction of the defects. The postoperative care of the patients is critically discussed. PMID- 12080624 TI - [Radioprotective effectiveness of amifostine on the salivary glands of the rat during fractionated irradiation]. AB - BACKGROUND: The aim of this study was to evaluate the cytoprotective potency of amifostine during a fractioned irradiation of salivary glands but also on the tumor response of rhabdomyosarcomas R1H in rats. METHOD: The head-neck-area of male WAG/RijH rats and the tumor in the flank were irradiated with 60Co-gamma rays (60 Gy/30 f/6 weeks). Amifostine (250 mg/m2) was applicated 15 min before irradiation. The control group was treated with an equal volume of physiologic sodium chloride. The salivary glands were exstirpated and examined histopathologically. Tumour volume was measured, the body-weight of rats determined. RESULTS: A dose dependent radiosialadenitis developed in all salivary glands. Amifostine had no influence on the response of R1H tumours to radiation. The animal weight loss in the amifostine group was higher as compared to control group. CONCLUSIONS: No cytoprotective effects on acute toxicity of salivary glands of rats could be detected under irradiation with synchronous application of amifostine. In addition, no effects on tumor response were observed. The general animal constitution decreased by additional medication of amifostine. PMID- 12080626 TI - [Expert assessment of oropharyngeal dysphagia]. AB - The number of patients with chronic dysphagia is increasing as more patients with neurologic diseases and carcinomas of the head and neck can be successfully treated and survive. The expert assessment of a dysphagia should be based not only on anamnestic data, as proposed by the German guidelines "Anhaltspunkte fur die arztliche Gutachtertatigkeit im sozialen Entschadigungsrecht und nach dem Schwerbehindertengesetz," but also on clinical, endoscopic, and radiologic examinations. Modern procedures of treatment enable us to prevent aspiration or insufficient nutrition; nevertheless, dysphagia causes severe limitations for the patients. PMID- 12080627 TI - [Papillary tumor of the inner ear: adenoma or carcinoma?]. AB - A 46-year-old female patient presented relapsed left-sided hearing loss and progressive left facial nerve palsy. At first, the computed tomography (CT) scan and magnetic resonance (MR) imaging did not indicate a diagnosis of a tumorous lesion. Nine years after the first symptoms, a destructive cystic neoplasm of the pars petrosa of the temporal bone was then evident. The lesion was surgically removed by radical resection. Histologically, a papillary tumor of the endolymphatic sac was diagnosed. These tumors tend to become destructive and more or less aggressive, but they do not metastasize. Therefore, the best term for them should be "aggressive papillary tumors of the endolymphatic sac" to avoid classification as an adenoma or adenocarcinoma. PMID- 12080625 TI - [Organ preservation in advanced laryngeal and hypopharyngeal carcinoma by primary radiochemotherapy. Results of a multicenter phase II study]. AB - INTRODUCTION: Regarding the promising results of international trials we conducted the first German prospective multicentre phase II trial for organ preservation with primary simultaneous chemoradiation in advanced laryngeal and hypopharyngeal cancer. PATIENTS AND METHODS: 28 of 30 recruited patients suffering from stage II and III (UICC) laryngeal and hypopharyngeal cancer were treated with primary simultaneous chemoradiation within an organ preservation program and monitored in follow-up of one year. Exclusion criteria included tumor infiltration of the laryngeal cartilage, bilateral neck nodes (N2c) and need for flap reconstruction in case of laryngectomy. The protocol included an accelerated concomitant boost chemoradiation (66 Gy) with Carboplatinum (70 mg/m2 1st and 5th week) and a restaging procedure one month after therapy. In case of residual disease, salvage laryngectomy and/or neck dissection were performed. RESULTS: After follow-up of one year 20 of 28 patients (71%) were presented with stable complete remission and functionally preserved larynx. Of these 20 patients 3 developed pulmonary metastases, 1 secondary primary carcinoma of the lung and 3 neck metastases which needed neck dissections. The other patients showed in 4 cases relapsing tumor which was indicated for laryngectomy. One patient needed tracheotomy because of persisting edema and 2 patients died due to tumor progress. One patient died after complications due to salvage surgery. CONCLUSION: The organ preservation protocol was feasible with well tolerated early toxicity. Problems of screening for recurrent disease, salvage surgery and late toxicity should be noted and pronounced in patient information. Further studies should focus on the improvement of patient selection which could be realized by induction Chemotherapy (using new components like taxan) and/or use of prediction factors such as tumor volume and hemoglobin levels. PMID- 12080628 TI - [Sinugenic metastasis of bilateral renal cell carcinoma (hypernephroma)]. AB - The paranasal sinuses are a rare location of metastases. The most frequent infraclavicular primary tumor is the renal cell carcinoma (hypernephroma). We report a case of a 64-old-woman with a bilateral renal cell carcinoma and metastases in the paranasal sinuses with destruction of the skull base. If symptoms such as progressive indolent periorbital swelling or intermittent epistaxis occur in adults, one should consider a metastasis of a renal cell carcinoma to the paranasal sinuses even 10 years after nephrectomy. PMID- 12080629 TI - [Lightening-induced injury of the air. Lightening-induced tympanic membrane perforation left with subsequent temporary pancochlear inner ear hearing loss as well as left-sided tinnitus]. PMID- 12080630 TI - [Clinical management of reflux laryngitis]. PMID- 12080631 TI - [Secretory radical cavity]. PMID- 12080632 TI - [Thrombosis induced by homocysteine does not depend on platelet aggregation or heparin cofactor's antithrombin activity]. AB - OBJECTIVES: To partly elucidate the relationship between homocysteine(Hcy) and thrombosis, we observed Hcy's effects on platelet aggregation and antithrombin activity of heparin cofactor in vitro. METHODS: Platelet aggregation assay and chromogenic assay were used in this research. RESULTS: Homocysteine had no effects on platelet aggregation induced by ADP and thrombin or heparin cofactor's antithrombin activity in vitro (P > 0.05). CONCLUSION: There is no correlation between Hcy and the "ADP passway" of platelet aggregation. Thrombosis induced by Hcy does not depend on heparin cofactor's antithrombin activity either. PMID- 12080633 TI - [Specific cellular and humoral immune responses induced by intramuscular injection of DNA vaccine containing HBV HBsAg gene in mice]. AB - To know the influences on the function of cellular and humoral immunity in mice by DNA vaccine containing HBV HBsAg gene, the DNA vaccine, pcDNA3-HBsAg, in which the HBsAg gene sequences of HBV were inserted into downstream of CMV promoter in pcDNA3, were injected intramuscularly into Balb/C mice. The results were that the serum anti-HBsIgG was detected in all of the pcDNA3-HBsAg immunized mice, but none in control mice; serum levels of Th1 cytokines, interleukine 2, and interferon gamma, were significantly higher in pcDNA3-HBsAg immunized mice than these in controls. It is indicated that the good responses of cellular and humoral immunity can be induced by injection of pcDNA3-HBsAg in Balb/C mice. PMID- 12080634 TI - [Protective effects of ischemic preconditioning on ischemia reperfusion injuries of kidney: experimental studies]. AB - We studied the serum levels of SOD and creatinine(Cr) of venous blood and changes in renal tissue structures under electromicroscope in the ischemia referfusion(I/R), the ischemic preconditioning(PC1, PC2, PC3), and the control groups in rabbits. The results were that the serum levels of SOD and Cr in the I/R group were obviously higher than that in the PC and control groups after one hour ischemia and one-hour reperfusion(P < 0.001). There were no significant difference in Cr levels between the PC2 and control groups; there were significant degenerate and necrotic changes under the electromicroscope in renal tissues in the I/R group, but there were no significant change in the PC and control groups. The results suggest that PC plays a role in protecting the kidney from injury induced by ischemia and reperfusion. PMID- 12080635 TI - [Ischemic preconditioning reduces lung ischemia reperfusion injury in vivo rabbits]. AB - Thirty male Japanese white rabbits were randomized into two groups. Fifteen rabbits were treated with ischemic preconditioning(their left lung hilus clamped for 10 min and released for 15 min; Group IP). Fifteen rabbits were not treated with ischemic preconditioning(Group C). Then the left lung hilus of both groups were occlused for 60 min and reperfused for 60 min. The results were that after 60 min of reperfusion, in Group IP, the contents of angiotensin II (Ang II) and artery oxygen tension(PaO2) in blood samples and the contents of superoxide dismutase in lung tissue were significantly higher, while the mean pulmonary artery pressure in blood samples, the wet/dry ratio and malondialdehyde content in lung tissue were significantly lower than those in Group C; Histological findings showed less damage in Group IP. It is suggested that lung ischemic preconditioning can reduce normothermic rabbit lung ischemia-reperfusion injury. PMID- 12080636 TI - [Experimental study of oxygen supply and demand balance during total hepatectomy and partial liver replantation with venous bypass technique in young pigs]. AB - OBJECTIVE: To investigate the changes of oxygen supply and demand balance during total hepatectomy and partial liver replantation(TCH and PLRP) with venous bypass technique in pigs. METHODS: Seven young pigs[weight (24.2 +/- 4.8) kg] were anesthetized with fentanyl and pentobartal. Catheters were inserted into portal vein via splenic vein and into vena cava inferior via right femoral vein, respectively, and the two catheters were connected by a "Y" adaptor to right atrium via extracorporeal circulation pump. Venous bypass technique was used during occlusion of total hepatic blood stream(OTHB). SvO2, hemodynamics and blood gas analysis were measured after anesthesia, during and 30 minutes after venous bypass. RESULTS: SvO2 was decreased significantly(P < 0.05), vein bypass technique ameliorated partially the disturbance of hemodynamics, and there was correlation between CO and SvO2 during THC and PLRP in pigs. CONCLUSION: The results suggest that there is oxygen supply and demand unbalance during THC and PLRP with venous bypass technique in pigs, and it is beneficial to monitor oxygen ladder during operation. PMID- 12080637 TI - [Regulation of p53 and bcl-2 proteins to apoptosis and cell proliferation in liver cirrhosis and hepatocellular carcinoma]. AB - In situ apoptosis labelling was used for detecting apoptotic cells, and immunohistochemistry for p53, bcl-2 proteins and proliferation cell nuclear antigen(PCNA) in hepatocellular carcinoma(HCC) and liver cirrhosis tissues. The results were that in HCC, the number of apoptotic cells was higher, the density of proliferation cells lower, and expressions of p53 and bcl-2 protein were stronger than that in liver cirrhosis, and they were related to differentiation degree of HCC. The data indicate that overexpression of bcl-2 and mutant p53 proteins, which causes imbalance between cell proliferation and apoptosis, may bring about genesis and development of HCC by selecting proliferation of cells. PMID- 12080638 TI - [Endogenous protective effects of superoxide dismutases on infectious brain injury in rats]. AB - We studied the alterations of MDA and three forms of SOD activities such as T SOD, CuZn-SOD, and Mn-SOD in rat cerebral tissues injected by bordetella pertussis (BP) to elucidate protective mechanism of SOD against the infectious brain injury. The results were that water content(WC), Evans blue content(EB), MDA, and Mn-SOD activities in 4 h and 24 h BP-treated groups increased and T-SOD and CuZn-SOD decreased compared to corresponding normal saline(NS)-treated groups, respectively(P < 0.01); MDA increased and had a positive correlation with WC and EB in 4 h BP treated group (r = 0.9650, r = 0.9441, P < 0.01, P < 0.01, respectively); Mn-SOD activities were elevated and had a negative correlation with WC, EB, and MDA (r = -0.8650, r = -0.9021, r = -0.9346, P < 0.01, P < 0.01, P < 0.01, respectively) in 24 h BP-treated group. The results suggest that the increase of component Mn-SOD activities may play an important role in vivo endogenous protective mechanism against delayed infectious brain injury. PMID- 12080639 TI - [Effects of danshen and shengmaiye on glomerulosclerosis by adriamycin in rats]. AB - OBJECTIVE: To investigate the effects of danshen(Salvia plectranthoides Griff.) and shengmaiye (Panax ginseng C. A. Mey, Ophiopogon japonicus Ker-Gawl and Schisandra chinensis Baill) on glomerulosclerosis induced by adriamycin in SD rats. METHODS: Left kidney of the animals was removed and after 7 days, adriamycin(6 mg.kg-1) was injected through tail vein, so as to establish an animal disease model. Then danshen or shengmaiye was injected to peritoneal cavity of the rats. All rats were killed by the end of the 8th week. Hemoglobin, BUN, cholesterol and protein in urea within 24 hours(ur Pro/24 h) were detected. IV collagen(IV col.) and laminin(LN) of renal cortex were determined by ELISA. The quantitation of IV col and LN in glomerular mesangial area was analyzed by computer pictures. RESULTS: Compared with model-danshen group(Group III), model shenmaiye group(Group IV) and normal control group(Group I), BUN, cholesterol(Ch) and ur Pro/24 h were increased obviously and hemoglobin(Hb) was decreased in model control group(Group II). The values of BUN, Ch and Ur Pro were lower in Group III and Group IV than those in Group II and higher than those in Group I (P < 0.05). The quantitation of IV col and LN within renal cortex and mesangial area was less in Group III and Group IV than that in Group II and more than that in Group I (P < 0.05 or P < 0.01). CONCLUSION: Dansheng and shengmaiye may play an important role in the treatment of glomerulosclerosis in rats. PMID- 12080640 TI - [Electrocardiographic T-wave amplitude of normal population and its correlation with age, sex and race]. AB - OBJECTIVE: To establish a normal electrocardiographic T wave amplitude standard in Chinese population according to age and sex. METHODS: The electrocardiographic T wave amplitudes in 4,322 persons among normal population; aged from birth to eighty-five years old divided into 11 age-groups, were analyzed. The 97.5th percentile(P97.5) and 2.5th percentile(P2.5) were considered as the upper and the lower limit of normal T wave amplitudes and T/R ratio. Wilcoxon rank sum test was used to determine whether the normal electrocardiographic T wave amplitude and the T/R ratio were different in P97.5 value versus mean + 1.96 s value, in men versus women, and in Chinese versus Americans. The linear correlation analysis was used to examine the association between normal T amplitude and age as well as between T/R ratio and age. RESULTS: The normal T wave amplitudes and T/R ratio in all leads were greater in P97.5 value than in mean + 1.96 s value and were greater in men than in women. The normal T wave amplitudes of adults were greater in Chinese than in Americans except V1 lead in which the amplitudes were greater in Americans than in Chinese. There was moderate or marked correlation of normal T wave amplitude and T/R ratio with age in most of the leads. CONCLUSION: There are marked differences of age, sex and race in normal electrocardiographic T wave amplitudes. PMID- 12080643 TI - [Use of transcranial Doppler in monitoring cerebral blood flow velocity during peri-operation]. AB - Transcranial Doppler was used to monitor 45 patients' middle cerebral artery blood flow velocity during anesthesia and cardiopulmonary bypass. The result was that the change in middle cerebral artery blood flow velocity was in correspondence with cerebral blood flow. This paper suggests that transcranial Doppler is a simple and practical instrument for monitoring cerebral blood flow. PMID- 12080642 TI - [Roles of sex hormones and oxygen free radical in coronary heart disease]. AB - To examine the roles of sex hormones and oxygen free radical(OFR) in coronary heart disease (CHD), the serum estradiol(E2), testosterone(T), SOD, MDA and lipid levels were measured in 44 postmenopausal women with CHD and 22 health women. Mean levels of T and MDA were significantly higher while mean levels of SOD and E2 were significantly lower in CHD group than those in control group. On relative analysis, there was a positive correlation between E2 and SOD, while a negative correlation was observed between T and MDA in CHD group. Eighteen postmenopausal women with CHD had been receiving nilestriol replacement therapy for 12 weeks. The serum SOD and HDL-C/TC levels increased significantly, but LDL-C and MDA decreased significantly. The overall data suggest that OFR plays an important role in CHD; Low E2 levels is a risk factor of CHD in postmenopausal women; Nilestriol replacement therapy can improve the metabolism of serum lipids and inhibit lipid peroxidation, therefore, it may be of benefit to postmenopausal women with CHD. PMID- 12080641 TI - [Expression of WT1 gene mRNA in leukemia]. AB - WT1 gene expression was determined in acute leukemic patients by RT-PCR. RESULTS: 19 patients were found to have positive results among 28 patients, the positive rate is 67.9%, and in 16 ANLL patients, 11 cases got positive results, in 12 ALL patients, 8 cases got positive results. It suggested that WT1 was highly expressed in acute leukemia. It is a new and significant detective marker for monitoring the effect of chemotherapy, predicting the prognosis and in the research of minimal residual disease (MRD). PMID- 12080644 TI - [Relation of left ventricular mass and QT dispersion in patients with essential hypertension]. AB - To study the relation between the rate-corrected QT interval dispersion(QTcd) and left ventricular mass, the blood pressure(BP), left ventricular mass index(LVMI), and QTcd were measured in 79 essential hypertensive patients. The results were that QTcd and QTcd% in hypertension with left ventricular hypertrophy(LVH) group were significantly higher than that in hypertension without LVH group; The higher the LVMI, the higher the QTcd and QTcd%(r = 0.7720, r = 0.6798, respectively), which were not associated with BP. The results suggest that LVH, but not hypertension, leads to an increase in QTcd. PMID- 12080645 TI - [Effect of desflurane on quantitative electroencephalogram and hemodynamics]. AB - To evaluate the feasibility of monitoring depth of desflurane anesthesia by quantitative electroencephalogram(QEEG) and hemodynamics, eight ASA I-II patients were studied. Anesthesia was induced by propofol and vecuronium, after tracheal intubation and the lungs were ventilated with desflurane in oxygen. The results showed that desflurane significantly reduced bispectral index(BI), 95% spectral edge frequency(SEF) and median frequency(MF) in a negative linear dose-related manner. When the end-tidal desflurane concentration reached 1.5 MAC, mean artery pressure(MAP) was decreased significantly, and heart rate(HR) increased by 14.5% (but there was no statistic significance), compared with pre-anesthesia. The results suggest that QEEG can be used to monitor depth of desflurane anesthesia, but MAP and HR are not the ideal monitoring indices during desflurane anesthesia. PMID- 12080646 TI - [Use of ultrasonography to evaluate carotid atherosclerosis and hemodynamics]. AB - One hundred and one cases of carotid atherosclerosis(CAS) were diagnosed by ultrasonography. The intermediate(IMT) of common carotid endomembrane in CAS group was much thicker than that in the healthy group(P < 0.01). There was no significant difference between the two groups in the peak velocity in systolic stage(MAX), the velocity in diastolic stage(MIN), resistance index(RI) and vessel size. The atheromatous plagues were usually flat plaques, seen at the bifurcation of common carotid artery. They belong to early pathological changes. It seems that the plaques don't affect MAX, MIN and RI of the common carotid artery. PMID- 12080648 TI - [Piggyback allogenic orthotopic liver transplantation: one case report]. AB - Piggyback allogenic orthotopic liver transplantation was performed successfully on one patient with primary liver carcinoma. The operation took 11 h and 30 min. The resection of the liver in the patient was initiated by the ligation and transection of the veins within the third hepatic portal. The hepatic venous system was reconstructed with the end of the superhepatic inferior vena cava anastomosed to the side of the inferior vena cava at the angle of 30 degrees-45 degrees in the recipient. The hemodynamics was steadfast intraoperatively, without extracorporeal venous bypass. The patient had a good post-operative recovery. The patient died of respiratory failure caused by metastasis. PMID- 12080647 TI - [Clinical research of ischemic preconditioning on lung protection]. AB - Twenty patients, who only need to occlude main pulmonary artery underwent pulmonectomy, were randomized into two groups. Ten patients were treated with ischemic preconditioning(IP). Their main pulmonary artery was clamped for 10 minutes and released for 10 minutes(Group IP). Ten patients were not treated with ischemic preconditioning(Group C). The results showed that the content of calcitonin gene related peptide(CGRP) in Group IP was significantly higher than that in Group C after ischemic preconditioning. After lung reperfusion for 30 minutes, the value of superoxide dismutase and pulmonary vein blood oxygen tension in Group IP was significantly higher than that in Group C. It is evident that ischemic preconditioning could reduce normothermic human lung ischemia reperfusion injury. The possible mechanism is that ischemic preconditioning increases the production of endogenous CGRP. PMID- 12080649 TI - [Relationship between brain edema induced by freezing injury and concentration of Evan blues and cytosolic free calcium in brain tissues]. AB - The authors studied the changes of Evan blues(EB) of brain tissues and concentration of cytosolic free calcium ([Ca2+]i), and relationship between them and water content of brain tissues following freezing brain edema in rats, to elucidate mechanism and type of brain edema induced by freezing injury. The results indicated that there were close positive relationship between water content and EB of brain tissues, and between [Ca2+]i and water content. We found that not only cytotoxic brain edema(intracellular edema) occurred but also occurred vasogenic brain edema(extracellular edema) followed by blood brain barrier breakdown after brain edema. Freezing brain edema is mixed brain edema. PMID- 12080650 TI - [Plasma Lp (a) levels and correlation of Lp (a) with fibrinolysis activation in chronic renal failure]. AB - Plasma Lp (a) concentration and activation of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1(PAI-1) were determined in 50 patients with chronic renal failure (CRF) and in 50 healthy subjects. The results demonstrated that plasma concentration of Lp(a) was significantly higher and plasma t-PA activation was significantly lower in CRF patients than in healthy subjects. With multivariate analyses, plasma Lp(a) concentration was positively correlated with the total amount of 24 h uric protein, plasma BUN and uric protein concentration. Moreover, a negative significant correlation between plasma Lp(a) levels and t-PA activation was observed. Plasma Lp(a) levels did not show a correlation to PAI-1 activation. It is indicated that renal function may contribute to moderate plasma Lp(a) concentration and elevated Lp(a) levels may be related with fibrinolytic impairment. PMID- 12080651 TI - [Single-catheter approach to radiofrequency current ablation of accessory pathway in Wolff-Parkinson-White syndrome with recurrent rapid atrial fibrillation]. AB - Twelve patients with WPW were treated by single-catheter ablation. Eight patients had the histories of recurrent rapid atrial fibrillation(Af) and refractory to antiarrhythmic agents. Ventricular rate varied between 250-300 bpm. Defibrillation was needed in three patients, and one had syncope. All patients were treated successfully by ablation of accessory pathway with single-catheter approach and no Af occurred during the procedures. This result suggests that the single-catheter ablation may be an effective and feasible treatment for these patients with WPW and may be helpful in avoiding Af in multiple-catheter introducing procedure. PMID- 12080652 TI - [Study of calcitonin gene methylation in chronic myeloid leukemia by using Hpa II PCR]. AB - To investigate the role of calcitonin(CT) gene hypermethylation in the transformation from the initial chronic phase to blast crisis of chronic myeloid leukemia, 31 CML patients were studied by using Hpa II-PCR. The results showed that the 10.52%(2/19) chronic phase, 71.4%(5/7) accelerated phase and 80.0%(4/5) blast crisis of patients had CT gene hypermethylation and that the increased methylation of the CT gene were related with the disease progression. The reports indicated that the hypermethylation of CT gene might be a useful marker for predicting the evolution of CML and selecting chronic phase patients for BMT. PMID- 12080653 TI - [Influence of nitric oxide upon cerebral infarction in 15 cases]. AB - To study the role of the nitric oxide(NO) in patients with cerebral infarction, NO in cerebrospinal fluid was determined in cerebral infarction group(15 cases) and control group(10 cases). Infarct volume was determined by CT between the second and the 7th day after the onset of symptoms. The severity of neurological deficits was assessed with the stroke scale of Ministry of Public Healthy(China). The results showed that NO was higher in cerebral infarction group (3.12 +/- 1.60) than that in control group(1.19 +/- 1.01) (P < 0.01). The concentration of NO was positively correlated with infarction volume(r = 0.57, P < 0.05) and severity of neurological of deficits(r = 0.54, P < 0.05), respectively. The results support the conclusion that NO produced in large amounts in the postischemic tissue contributes to the progression of the brain damage, which was demonstrated in animal models of focal cerebral ischemia. PMID- 12080654 TI - [Effect of glycyrrhizin on plasma endothelin and serum nitric oxide in myocardial ischemia reperfusion rabbits]. AB - Effects of glycyrrhizin on plasma endothelin(ET), serum nitric oxide(NO) and creatine phosphokinase(CPK) activity of myocardial tissues were observed in myocardial ischemia/reperfusion rabbits. The results indicated that the CPK activity and plasma ET level increased and serum NO level decreased significantly after myocardial ischemia/reperfusion. In comparing with control, glycyrrhizin not only reduced significantly the increased plasma ET level and CPK activity of myocardial tissues, but also increased markedly the producing and releasing of NO(P < 0.01). All these facts suggest that glycyrrhizin exerts obviously protective effects on myocardial injury induced by ischemia/reperfusion. PMID- 12080655 TI - [Relationship of nitric oxide and immunal function of the patients with acute hepatitis and hepatic cirrhosis]. AB - The interleukin-12(IL-12), interleukin-10(IL-10), tumor necrosis factor(TNF) and nitric oxide (NO) levels in serum of 90 patients with hepatitis and cirrhosis were measured by the method of ELISA and colorimeter. The levels of IL-12, IL-10 and NO in serum of the patients with hepatitis were in close proximity to the level of healthy control, but the TNF level was significantly higher than that of healthy control. The IL-12, IL-10, TNF and NO levels in serum of the patients of hepatic cirrhosis were significantly higher than that of healthy control and patients with hepatitis. The results suggested that the NO level was not related with the hepatic damage of acute hepatitis. The dysfunction of immunology in the patients with cirrhosis was related with the increase of NO level. PMID- 12080656 TI - [Study on the relationship between plasma endothelin nitric oxide concentration and renal hypertension and renal function]. AB - OBJECTIVE: To investigate the relationship between plasma endothelin(ET), nitric oxide(NO) levels and, renal hypertension and renal function. METHODS: The plasma concentration of ET-1 was detected by immunofluorescence assay. The plasma concentration of NO was detected by biochemistry assay. RESULTS: 1. In renal disease patients, plasma concentration ET-1 was markedly elevated, and plasma concentration of NO was decreased, compared with the healthy subjects(P < 0.01). 2. Plasma concentration of ET-1 was markedly increased and plasma concentration of NO was decreased in the patients with renal hypertension. 3. Plasma level of ET-1 was higher, and plasma level of NO was lower in the patients with renal function damage than that of those without renal function damage. 4. BP, BUN and Scr were positively correlated with plasma ET-1, but they were negatively correlated with plasma concentration of NO. CONCLUSION: Plasma ET-1 and NO may play an important role in pathogenesis of renal hypertension; the change of their levels may be related to the progress of these renal diseases. PMID- 12080657 TI - [Study on combined clinical manifestation and myocardial perfusion imaging with 99mTc-MIBI SPECT for diagnosing myocarditis]. AB - In 568 cases suffering from upper-respiratory tract infection with sore throat and more than one of following clinical manifestations: arrhythmia, heart failure and chest pain or oppressive sensation over the chest(Group A), and another 108 cases without above clinical manifestations(Group B), myocardial perfusion imaging with 99mTc-MIBI SPECT was studied. The results revealed that the imaging in 404 cases (71.12%) of Group A supported diagnosis of myocarditis, but only 6 cases(5.56%) did so in Group B. Because of lacking the clinical manifestations of myocarditis but positive finding of CVB-IgM antibody was detected in their sera, we considered that the positive SPECT imaging in these six cases of Group B belonged to myocardial reaction due to virus infection. It was possible that some of them were suffering from subclinical type of myocarditis. PMID- 12080658 TI - [Measurement of cyclosporine A in whole blood by high performance liquid chromatography(HPLC)]. AB - Whole blood was extracted by ether under base using cyclosporine D as internal standard. The extraction was evaporated and reconstituted. The interferences were further removed by hexane. The reconstitution was injected into a C18 reversed phase column at 70 degrees C, which was eluted with a mobile phase of acetonitrite/methanol/water(60/10/30, v/v) at flow rate 1.6 ml.min-1 and monitored at 210 nm. Three levels of BIO-RAD cyclosporine A(whole blood) calibrator were measured, all the results were within expected range. The detection limit was 25 ng.ml-1. The linearity was at least within 50-1,000 ng.ml 1. Analytical recoveries were 99.6%-108.8%. The within-run cv and between-run CV were 7.2% and 9.3%, respectively. PMID- 12080659 TI - [Automatic hemacytometer applied for detecting white blood cell in the patients with severe jaundice hepatitis]. PMID- 12080660 TI - [Investigation of human herpes virus-6 antibodies of adults in Changsha region]. PMID- 12080661 TI - [Evaluation of rapid detection for serum lipoarabiomannan-IgG in the diagnosis of tuberculosis]. PMID- 12080662 TI - [Analysis of re-establishing cardiopulmonary bypass after neutralization of heparin during open heart surgery]. PMID- 12080663 TI - [Changes in nitric oxide and endothelin on experimental guinea pigs with asthma]. PMID- 12080664 TI - [Forensic clinical analysis of sciatic nerve injury induced by drug injection through buttock muscle]. PMID- 12080665 TI - [Monitoring the acute rejection after lung transplantation by flow cytometry]. PMID- 12080666 TI - [A case of arrest of erythrohematopoiesis]. PMID- 12080667 TI - [Chinese hamster emotionality rating scale]. AB - To establish the rating method of Chinese Hamster emotionality and verify its reliability of assessment result so as to investigate the role of emotionality in onset/incidence of diabetes in Chinese Hamster of non-insulin-dependent diabetes mellitus, Ader's rating methods of emotionality were revised, the level of emotionality for Chinese Hamster was measured, cut off score of emotionality was computed, and reliability of rating scale was analysed. The results were that the cut off score of emotionality which was 5 scores could divide the high/low emotionality response in individual animal; Some items were moderately and highly correlated to total score (r = 0.3268-0.7347); The test-retest reliabilities of total score after 1 and 8 days ranged 0.5874 to 0.6105, the inter-rater agreement for reliability trials was 95.2%, and its Kappa value was 0.905. The results suggest that Chinese Hamster Emotionality Rating Scale has the satisfactory reliability. PMID- 12080668 TI - [The effects of muscarinic and nicotinic receptor antagonists on compound action potentials in the cochleae of gerbils]. AB - To explore the mechanism of the medial olivocochlear(MOC) efferents and their neurotransmitter acetylcholine in regulating the cochlear mechanics and the auditory afferent activities, the effects of contralateral noise(CLN) and muscarinic(M) and nicotinic(N) receptor antagonist on the compound action potential(CAP) of the cochleae of gerbils were observed in our studies. The results showed that the CAP was significantly inhibited by CLN, and the suppressive effect of CLN on CAP was eliminated either by atropine or by gentamycin. These results support that MOC efferents play their physiological effect via both M and N receptors. PMID- 12080669 TI - [Study on the prevention of insulitis and diabetes by complete Freund's adjuvant in NOD female mice]. AB - The degrees of insulitis, the incidence of diabetes, and the counts of alpha,beta and delta cells in islets were studied by morphological method and avidin-biotin complex immunohistochemical method in complete Freund's adjuvant(CFA) or normal saline(NS)-treated non-obese diabetic(NOD) female mice. The results showed: The incidences of in-insulitis and the scoring means were significantly lower in CFA treated mice than those in NS-treated ones; none of 5 CFA-treated mice and 3 out of 5 NS-treated ones developed diabetes. The positive rates of alpha, and delta cells were significantly lower in CFA-treated mice than those in NS-treated ones, but the positive rate of beta cell was significantly higher in CFA-treated mice than that in NS-treated ones. The correlations were found among the scores of insulitis and the positive rates of alpha, beta, delta cells in islets (r alpha = 0.475, r beta = -0.878, r delta = 0.869). The results indicate that CFA may lessen the degrees of insulitis and the incidence of diabetes. Its effects might be related to lessen the damage of beta cell in islets. PMID- 12080670 TI - [Apoptosis of chondrocytes in experimental osteoarthritis]. AB - The right knees of rabbits were immobilized in full extension for up to eight weeks using plaster cast. Specimens of the articular cartilage obtained from tibial plateau were studied by histopathologic and TDT-mediated fluorescein-dUTP nick-end labelling(TUNEL) techniques. The results showed that TUNEL-positive chondrocytes with apoptosis specific morphology were detected on superficial and middle layer of the articular cartilage from one to two weeks after immobilization, and these changes progressed until 4 weeks after immobilization. Six weeks after immobilization, TUNEL-positive chondrocytes were seen through the entire thickness of the articular cartilage. Our findings indicate that apoptosis of chondrocytes could be induced by immobilization and might be responsible for articular cartilage degeneration, and which is one of the pathways involved in the pathophysiological mechanism of osteoarthritis. PMID- 12080671 TI - [Detection of bcr/abl gene expression on bone marrow cell colonies in chronic myelogenous leukemia by reverse transcriptase-polymerase chain reaction]. AB - The t(9;22) (q34;q11) between abl and bcr genes plays a pivotal role in the diagnosis and pathogenesis of chronic myelogenous leukemia(CML). To explore the bcr/abl fusion mRNA expression on hematopoietic precursors, reverse transcriptase polymerase chain reaction(RT-PCR) was applied to detect bcr/abl mRNA expression on bone marrow cell colonies. Meanwhile, bcr/abl mRNA expressions on 14- or 28 day colonies using HPP-CFC and CFU-GM semisolid agar culture assay were also determined in 4 cases of confirmed Ph-positive CML by karyotyping analysis. The results showed that the bcr/abl mRNA expressions on 14-day colonies and some 14- or 28-day colonies detected singly were positive at presentation by RT-PCR, in agreement with results by karyotype analysis. Thus, a sensitive and powerful technique was offered for studying gene expression on hematopoietic precursors, diagnosis and therapeutic monitoring of CML. Furthermore, this can be used as an ideal method for revealing molecular mechanisms of pathogenesis of CML and screening anti-CML drugs. PMID- 12080672 TI - [Observation of apoptosis and proliferation in nasopharyngeal carcinoma]. AB - In order to observe the apoptosis and proliferation of cells in nasopharyngeal carcinoma(NPC) and pericarcinomatous tissue(PCT), 49 cases of NPC and partly PCT were detected by in situ end-labelling (ISEL) technique and proliferating cell nuclear antigen(PCNA) immunohistochemical staining. The results showed that: 1. Massive distribution of ISEL positive cells was frequently exhibited in vesicular nucleus cell carcinoma(VNCC) of NPC, in which no apoptotic pattern character was found. 2. The positive rate and the staining intensity index(SII) of ISEL signals in NPC were higher than that in PCT(P < 0.05); the positive correlation between ISEL SII and PCNA SII was found in NPC(r = 0.341, P < 0.05), whereas the positive rate and the SII of ISEL signals were significantly lower than that of PCNA (P < 0.01). 3. The ISEL SII and the PCNA SII in VNCC were obviously higher than that in poorly differentiated squamous cell carcinoma(PDSCC) (P < 0.01, P < 0.05). Patients with VNCC had a high five-year survival rate. The results suggest that there is a relationship between cellular proliferation and cellular apoptosis in NPC. PMID- 12080673 TI - [Analysis of renal function on 1,743 people of different ages]. AB - OBJECTIVE: To understand and evaluate the currency and correlation between parameters about renal function. METHODS: We divided 1,743 people whose ages vary from 20-85 years into six groups(A, B, C, D, E, F). Urinary Mini Albumin, RBP, NAG, osmotic pressure, serum BUN, plasma UA and internal creatinine clearance were measured as parameter about renal function. RESULTS: There was no obvious difference between the Group A and Group B(P > 0.05); The mean value of urinary NAG and urinary RBP in the two group was 10.9650 +/- 6.5650 and 0.1885 +/- 0.4709; However, when they were compared with the values of C, D, E group respectively, there were prominent differences (P < 0.05 or P < 0.01). Urinary Alb value of Group A is quite different from other groups. No difference exists between urinary Alb value of Group E and Group F(P > 0.05). Internal creatinine clearance decrease by the rate of 8.5%, 17.8%, 29.9%, 42.9% and 56.3% when Groups B, C, D, E and F was compared with Group A; while urinary osmotic pressure and the value of plasma BUN, Cr and UA has no obvious difference between any two groups(P > 0.05). CONCLUSION: The clearance of internal creatinine decrease with the growing of the age, especially older than 50 years of age. The parameter about tubular injury increase obviously after 40 years old though it does not change prominently before 50 years old, while it tends to be stable after 70 years old. We think it is during period of 40 to 70 years old that the renal function decreases more rapidly. PMID- 12080674 TI - [Effects of plasma catecholamines contents on acute cerebral hemorrhage treated with nao yi-an granule]. AB - In order to investigate the efficacy mechanism of Nao Yi-an(NYA) granule on acute cerebral hemorrhage(ACH), 38 cases of ACH were treated with NYA granule(NYA group) or with Western Medicine (WM group). The contents of their plasmal NE, E, and DA before and after treatments were observed. The results showed that the contents of the plasmal NE, E and DA increase remarkably, compared with those of normal control group(P < 0.01), and accompanied by the increase of volume of hemorrhage or the gravity of the disease. After treatment, the contents of the plasmal NE, E and DA in NYA group are notably lower than those before the treatment(P < 0.01), but in WM group, they are not significantly lower than those before the treatment(P > 0.05), and they are strikingly lower in NYA group than in WM group. It is indicated that the NYA group can effectively decrease the contents of plasmal NE, E and DA, to remove cerebrovascular spasm, to relieve cerebral edema, to improve microcirculation of brain tissue, to increase blood and energy supplies, anticerebral secondary ischemia and hypoxia and improve the clinical effects. PMID- 12080676 TI - [Cerebral hemodynamic impairment and therapeutic effect of external counterpulsation on elderly patients with brain infarction]. AB - Hemodynamic parameters of middle cerebral artery(MCA) were measured with Transcranial Doppler (TCD) in 31 elderly patients with brain infarction during convalescence and 28 healthy elderly controls. Twenty-two elderly patients with brain infarction and cerebral hemodynamic impaired were treated by External Counterpulsation(ECP). The results showed that the Vp and Vm of MCA in patients were significantly lower than that in the healthy controls(P < 0.001), the DVp, DVm and asymmetry of MCA in patients were greater than that in the healthy elderly(P < 0.001, P < 0.005). The Vp and Vm of MCA in patients after ECP were significantly higher than before(P < 0.001). The DVp, DVm and asymmetry of MCA in patients after ECP were significantly lower than before(P < 0.001, P < 0.005). The hemodynamic therapeutic effect is 81.8%. PMID- 12080675 TI - [Efficacy of tianlong xifeng granule (TXG) in the treatment of acute cerebral infarction of ganyanghuafengzheng]. AB - In this article 412 patients with acute cerebral infarction of ganyanghuafengzheng are analyzed in order to compare the efficacy between TXG and xixian tongshuan pill(XTP). The patients were randomly divided into two groups: 305 patients in Group I were treated with TXG, and 107 patients in Group II treated with XTP. The results showed that the curative rate in Group I was 69.84%, while in Group II 46.73%. It is indicated that the efficacy of TXG is superior to that of XTP. No toxicity and side effect were found in the above two groups. PMID- 12080677 TI - [Study on regulators of the cell cycle in cutaneous squamous cell carcinoma]. AB - In order to investigate the relationship between the regulators of cell cycle and cutaneous squamous cell carcinoma(SCC), we used an immunohistochemical staining technique to examine the expression of cyclin D1, p16 and Rb in the paraffin embedded skin cancer tissues of 30 patients with SCC. The positive percentage of cyclin D1, CDK4, p16 and Rb was 66.7%, 53.3%, 33.3%, and 36.7%, respectively. The overexpression of cyclin D1 and CDK4 could be observed frequently in low grade of differentiated cells, especially in marginal cells of the cancer nest. The results indicate that the aberrant expression of positive and negative regulators of the cell cycle may be involved in the carcinogenesis and evolution of SCC. PMID- 12080678 TI - [Effects of apoptosis of synovial lining cells on pathogenesis of osteoarthropathy]. AB - The author counted synovial lining cells(SLCs) of apoptosis in the joints of osteoarthropathy(OA, 12 cases) and control(10 cases) patients. The counts of apoptotic SLC were investigated by gel electrophoretic analysis of DNA extracted from synovium. The apoptotic index(AI) of SLCs in OA(2.04%) was obviously higher than that in controls(0.60%); electrophorectogram of DNA revealed a characteristic ladder pattern of DNA fragments at 200 bp in the OA group. We conclude that apoptosis can be found in synovial lining layer, and the AI of SLCs in the OA group is much higher than that in the control group, implying the high turn-over rate of SLCs in OA; the increasing apoptosis of SLCs may increase lesion factors retained in joints and cause the injury of articular cartilage. PMID- 12080679 TI - [A role of nitric oxide and endothelin in bronchial hyperreactivity in patients with mild and middle asthma]. AB - OBJECTIVES: To probe into a role of nitric oxide(NO) and endothelin(ET) in bronchial hyperreactivity(BHR) in patients with mild and middle asthma. METHODS: MchPC20 was used as an index to evaluate BHR in the asthmatic patients. Endothelin and NO level in bronchial alveolar lavage fluid(BALF) obtained from 15 patients with attacking asthma(Group I), 8 patients with stable asthma (Group II) and 7 healthy control subjects (Group III) were examined by radioimmunoassay and copper coated cadmium reduction method, respectively. RESULTS: 1. MchPC20 and FEV1% in Group III were slightly higher than those in Group I (P < 0.05), and lower than those in Group III (P < 0.05), respectively; 2. NO and ET levels in BALF from Group I were higher than those from Group II and Group III (P < 0.05), respectively; 3. ET/NO ratio in Group I was higher than that in Group III (P < 0.05) and lower than that in Group II (P > 0.05); 4. NO and ET level in BALF from Group I were positively and negatively correlated with MchPC20, respectively(r = 0.83, P < 0.01; r = 0.75, P < 0.01); the correlation of NO and ET level in BALF from Group I was negative (r = 0.70, P < 0.01); 5. NO and ET levels in BALF and ET/NO ratio from Group II were slightly higher than those from Group III (P < 0.05), respectively. CONCLUSION: NO may improve and ET exacerbate BHR in patients with mild and middle asthma. Bronchial hyperreactivity of the patients with stable asthma displays lightly abnormal, which may be due to mild abnormality of NO, ET levels and ET/NO ratio in BALF. PMID- 12080680 TI - [Significance of the changes of plasma endothelin, norepinephrine and atrial natriuretic peptide in treatment of heart failure with beta-blocker]. AB - Thirty patients with chronic heart failure after conventional treatment for two weeks with cardiac diuretic and vasodilator were divided into two groups. There was addition of betaloc into Group A, but Group B kept the original treatment plan for 8 weeks. Group C consisted of 16 normal controls. The results showed that: 1. The level of plasma endothelin(ET), norepinephrine(NE), atrial natriuretic peptide(ANP) in Group A and B were higher than those in Group C. 2. The decreasing degrees of ET, NE, ANP, heart function score and the increasing degrees of CI and LVEF in Group A were more than those in Group B. 3. The change of ET was related to the changes of the others. Therefore, the beta-blocker may be associated with the decrease of the plasma concentration of ET, NE and ANP and improve heart function. It is considered that the degree of change of plasma ET may be the simple and reliable index which evaluate the degree of improving heart function. PMID- 12080681 TI - [Changes in lipoprotein (a) and the relation between lipoprotein (a) and endothelin in patients during upper abdominal surgery]. AB - OBJECTIVE: We observed the changes in both serum lipoprotein(a) [Lp(a)] and plasma endothelin (ET) during upper abdominal surgery under intravenous combined with inhalation anesthesia and we also analyzed their correlativity. METHODS: Fifteen patients, scheduled for upper abdominal surgery, were anesthetized with total does of fentanal 8-12 micrograms.kg-1 and inhalation of 1-2 MAC enflurane. Serum Lp(a) and plasma ET at pre-anesthesia, pre-operation, 1 h from the start of operation and 30 min after operation were determined. RESULTS: The average concentrations of Lp(a) and ET were increased at 1 h from the start of operation compared with pre-anesthesia, but had no statistical significance. Lp(a) and ET were significantly higher at 30 min after operation than pre-anesthesia(P < 0.05). CONCLUSION: Lp(a) and ET have a close relation in advancing cardio cerebral blood vessel diseases. PMID- 12080682 TI - [Changes in p50 and oxygen metabolism during mild hypothermia cardiopulmonary bypass]. AB - OBJECTIVE: To study the changes of p50 and oxygen metabolism during mild hypothermia cardiopulmonary bypass, arterial blood lactate(ABL) were measured, arterial and venous blood gas were analysed in 21 patients undergoing cardiac valve replacement. RESULTS: p50, oxygen consumption(VO2), and oxygen extract rate(ERO2) were decreased during hypothermia and increased during rewarming, respectively (P < 0.05 or P < 0.01), while the change of SvO2 was adversed. PvO2 was decreased significantly during rewarming(P < 0.01), and ABL was continuously increased. CONCLUSION: Both reduction in oxygen utilization during hypothermia and relative deficiency of oxygen supply during rewarming cause tissue hypoxia. PMID- 12080683 TI - [Clinical and pathological analysis in 327 cases with renal biopsy]. AB - The renal biopsy results of 372 cases are summarized in this article. Definite pathological diagnoses were made in 354 cases, among which most cases were of mesengioproliferative glomerulonephritis type (27.1%), followed by glomerulopathy type with minimal change(12.4%). The association between clinical manifestation and pathological change was discussed. The data indicate that renal biopsy plays pivotal role in confirming the clinical diagnosis and guiding treatment. PMID- 12080684 TI - [Investigation on acute stroke patients being admitted to hospital]. AB - One hundred and twelve patients with acute stroke were studied. The results revealed that about 42 percent of 112 patients could get to hospital within 6 hours after onset, in which included 60.4 percent of the hemorrhagic group and 28.1 percent of the infarction group. Comparatively, among 30.3 percent of 112 patients CT scan was carried out within 6 hours, which included 41.7 percent of the hemorrhagic group and 21.8 percent of the infarction group. Linear correlation analysis was studied between admission time(AT) and the assessment of neural function defect(ANFD). The results showed that there was significant negative correlation between AT and ANFD in stroke patients. After analysing the serial reasons of delaying hospitalization, the authors have found that the key factor is the ignorance of the importance of stroke in early stage. PMID- 12080685 TI - [Clinical observation on ondansetron in preventing nausea and vomiting after cholecystectomy under abdominoscope]. AB - OBJECTIVE: To evaluate the prophylactic effect of ondansetron on postoperative nausea and vomiting (PONV) of cholecystectomy under abdominoscope. METHODS: Sixty patients, scheduled for selective cholecystectomy under abdominoscope, were randomly and double-blindly allocated to receive an intravenous bolus of either 0.9% NS 10 ml(Group placebo, n = 30) or 8 mg of ondansetron in 10 ml solution(Group ondansetron, n = 30), respectively. RESULTS: The incidences of nausea and vomiting in ondansetron group(33.3% and 26.7%) were significantly lower than those in placebo group(73.3% and 66.7%) (P < 0.05). Administration of ondansetron had no obvious effect on the mean arterial pressure, SpO2, and respiratory frequency. CONCLUSION: Ondansetron can be used effectively and safely to prevent PONV in cholecystectomy patients under abdominoscope. PMID- 12080686 TI - [Protective effect of Radix Salvia Miltiorrhizae on radiation damage of the cochlea]. AB - In order to evaluate the preventive and therapeutic effects of Radix Salvia Miltiorrhizae on radiation damage of the cochlea, guinea pigs were divided into 3 groups. Group 1 was treated by radiation added with Radix Salvia Miltiorrhizae. Group 2 was radiated alone, and Group 3 was control. Group 1 and Group 2 were radiated with a single dose of gamma radiation(60 Gy). Morphological and functional observation of the cochleae was performed in two weeks after radiation. The result showed that the changes of complex action potential(CAP) and the cochlear structure were slight in Group 1, but obvious changes were found in Group 2. There was a significant difference in CAP response threshold and incidence of the cochlear hair cell loss between Group 1 and Group 2 (P < 0.01). These results suggest that Radix Salvia Miltiorrhiza may prevent radiation induced cochlea damage. Its protective mechanisms may be cleaning free radicals, blocking calcium channel and improving microcirculation of the cochlea. PMID- 12080687 TI - [Perhydrit and sodium bicarbonate improve maternal gases and acid-base status during the second stage of labor]. AB - Perhydrit and sodium bicarbonate were administered to 99 women during the second stage of labor. Gas analysis was done twice at the beginning and the end of the second stage. The results showed that 33 women received both perhydrit and sodium bicarbonate(Group I) had significantly higher pH, BE, pO2, and O2sat at the end of the second stage than those of 30 controls (Group III) (P < 0.05 and < 0.01). And 33 women received only sodium bicarbonate(Group II) showed merely higher pH and BE than those of Group III. The difference was significant(P < 0.05 and < 0.01), while PO2 and O2sat had no statistical difference between Group II and Group III. By comparing the values, we found that pH, PO2 and O2sat were much higher at the end of the second stage in Group I and Group II than those at the beginning of the second stage(P < 0.05 and < 0.01). No statistical difference of BE had been found between both groups. In contrary, BE, pH, PO2 and O2sat declined at the end of the second stage in Group III (P < 0.02 and < 0.01). There were no significant differences of PCO2 among the three groups. It is suggested that the administration of perhydrit and sodium bicarbonate may effectively improve the tendency of maternal metabolic acidosis and tissue oxygen debt caused by consumption and uterine contractions during the second stage, subsequently improve maternal gases and acid-base status. PMID- 12080688 TI - [Changes in calcium and magnesium levels of lymphocytes in patients with diabetes and hypertension]. AB - To investigate the cellular basis linking non-insulin-dependent diabetes mellitus(NIDDM) and hypertension, we used the atomic absorption spectrometry to detect Ca2+ and Mg2+ levels of serum and lymphocytes in patients with the above diseases. The results were that serum Ca2+ and Mg2+ levels had no change, but lymphocyte Ca2+ level increased obviously and lymphocyte Mg2+ level decreased obviously in both diseases, compared with normal controls. There were no difference between hypertension and NIDDM with and without hypertension. Ca2+ and Mg2+ levels of lymphocytes in patients with hypertension correlated with the systolic pressure respectively(Ca2+, r = 0.4633, P < 0.05; Mg2+, r = -0.4412, P < 0.05), but their levels in patients with NIDDM were not correlated with the fasting blood glucose. The results suggest that there is an abnormal Ca2+ and Mg2+ metabolism in lymphocytes in patients with hypertension and NIDDM. PMID- 12080689 TI - [Clinical observation and experimental study on the leukopenia treated with shengbaikuai granule]. PMID- 12080690 TI - [Ultrasound image in different locations of the intestinal obstruction]. PMID- 12080691 TI - [Protective effects of ischemic preconditioning on myocardium during patients undergoing heart valve replacement surgery]. PMID- 12080692 TI - [Relation among some cytokines, nitric oxide and the changing condition of cirrhosis of liver]. PMID- 12080693 TI - [Otoendoscope in the assistant treatment of chronic suppurative otitis media]. PMID- 12080694 TI - [Radioimmunoassay of serum vitamin B12 and folic acid in patients with pernicious hemopathy]. PMID- 12080695 TI - [Diagnosis of congenital unilateral ectopic small kidney with ectopic ureteral orifice by color ultrasonography]. PMID- 12080696 TI - [A report of mini-thoracotomy in 12 patients]. PMID- 12080697 TI - [Hysterectomy in a case of normal pregnancy complicated with brain tumor]. PMID- 12080698 TI - [Gastroduodenal ulcer, perforation after chemo-embolization in 4 cases of advanced hepatocellular carcinoma]. PMID- 12080699 TI - [Application of color Doppler ultrasonography for assessing renal hemodynamics in the patients with hepatic cirrhosis and portal hypertension]. PMID- 12080700 TI - [Tubulation of peripheral nerve defects of upper limb using silastic tube]. PMID- 12080701 TI - [Effect of ischemia-reperfusion injury on the expression of nitric oxide synthase in rat retina]. PMID- 12080702 TI - [A case of refractory hypertension complicated with thoracic and pericardial effusion by peritoneal dialysis]. PMID- 12080703 TI - [Effect of long-term experimental stress on blood glucose level in mice with diabetes-prone: the age difference and its mechanism]. AB - OBJECTIVE: To compare the effects of long-term experimental stressors on blood glucose level and insulin secretion in the aged and young mice with diabetes prone and to explore the mechanism. METHODS: Diabetes-prone was induced by streptozotocin (STZ) in 40 1.5-month-old mice and 40 15-month-old mice. The young and the aged STZ mice were divided randomly into the stress group and the control group, respectively. Each group contained 20 mice. Both the aged and the young stress mice were exposed to multiple stressors (restrain, rotation, crowding) for 6 weeks. Fasting blood glucose (FBG), fasting plasma insulin (FINS), pancreas lipid peroxidase (LPO) and the metabolic products of nitric oxide (NO2-, NO3-) levels and superoxide dismutase (SOD) activity were measured. The grade of insulitis in STZ mice was also evaluated. RESULTS: After 6-week stress, FBG level, pancreatic tissue LPO and NO2-, NO3- contents were significantly higher while FINS level and SOD activity lower in the aged stress mice than those in the youngs. The insulitis malignization in the aged stress group was severer than that in the young stress group. There were correlativities among the above variables. CONCLUSION: The effects of long-term stress in promoting hyperglycemia exacerbating insulitis, and inhibiting insulin secretion are more significant in the aged STZ mice than in the young ones. This difference may result from the excessive NO and the peroxide injury to the pancreas. PMID- 12080704 TI - [Cloning of a new cDNA: responded to all-trans retinoic acid in HL-60 cell differentiation]. AB - In this paper a new type of cDNA fragment named as W-1 gene was first cloned and sequenced from ATRA induced HL-60 cells by using differential display PCR (DD PCR) and DNA sequencing techniques. These differentially expressing products of the gene responding to ATRA were further confirmed by Northern blotting analysis. The results showed that the expressing level of this gene induced by ATRA (10(-6) mol.L-1) for 16 hrs was much higher than that in the control HL-60 cells, but its expression in HL-60 was reduced to an unestimable level after induction of ATRA (10(-6) mol.L-1) for 24 hrs. It suggests that ATRA may increase the expression of W-1 gene during the early stage of HL-60 cell differentiation. The exact mechanism of action is being studied further. PMID- 12080705 TI - [Changes in trace elements in epithelial cells of oral submucous fibrosis]. AB - OBJECTIVES: To assay the concentrations of the trace elements in intranuclei and extranuclei between oral submucous fibrosis (OSF) and normal buccal membrane, to observe the relationship between the trace elements and OSF, and finally to understand the mechanism of OSF. METHODS: The concentrations of the trace elements of internuclei and extranuclei in OSF epithelial cells were assayed in computer automatic process by electron probe micro-analysis. RESULTS: The concentrations of Cu, Zn, S, and As in OSF were significantly lower than those in the normal buccal membrane (P < 0.01). The concentrations of Se and Mo were significantly higher than those in the normal buccal membrane(P < 0.01). CONCLUSION: The epithelial cells of OSF are seriously damaged by the low level of Cu and Zn and high level of Se in intracell. These changes are the element bases that OSF has canceration trend. PMID- 12080706 TI - [Study on antifungal mechanism of alpha-pinene]. AB - The antifungal mechanism of alpha-pinene was studied using electronic microscopy and incorporation of isotope-labelled precursor. The results demonstrated that alpha-pinene had significant roles in inhibiting and killing Candida albicans. After treatment with alpha-pinene the fungal morphology and ultrastructure showed obvious changes: their cell wall and cytoplasmic membrane ruptured; intracellular components released out and the cell residue fused to form irregular masses. In addition, the synthesis of DNA, RNA, polysaccharide of cell wall and ergosterol of cytoplasmic membrane was inhibited. It is indicated that these changes are related to antifungal mechanism of alpha-pinene. PMID- 12080707 TI - [Effect of pertussis bacilli on the release of excitatory amino acids in brain slice of rats]. AB - OBJECTIVE: To investigate the effect of pertussis bacilli on the release of excitatory amino acids (EAAs) in rat-brain slice and to know whether MK 801(dizocipine) protects brain tissue from excitotoxicity. METHODS: SD rats were randomly divided into 3 groups: 1. Normal slice cultured group(NC); 2. 10% pertussis bacilli(PB) slice cultured group(PBC); 3. 10% PB slice cultured group after pretreatment with MK-801. The release of EAAs from brain slices was measured by high performance liquid chromatography. RESULTS: The concentrations of glutamate(Glu 29.53 +/- 8.00), aspartate(Asp 23.74 +/- 5.79) and LDH(38.03 +/- 8.45) were significantly higher in PBC group than those(Glu 16.46 +/- 4.00, Asp 15.27 +/- 5.31, LDH 23.10 +/- 5.11) in MPBC group and (Glu 16.09 +/- 3.74, Asp 11.47 +/- 2.51, LDH 25.82 +/- 4.50) NC group. CONCLUSION: Pertussis bacilli may elicit a large increase of extracellular EAAs and LDH in vitro, while MK-801 protects brain tissue from excitotoxicity. PMID- 12080708 TI - [Influence factors in the clearance of urea and creatinine through gastrointestinal tract]. AB - To investigate the affection of different osmotic density, pH value, and dialytic fluid with drugs on the clearance of urea and creatinine through gastrointestinal tract, we used the renal-artery-ligated rabbits as the model. After the intravenous administration of urea and creatinine through ear vein, fistulas in duodenum at about 5 cm to pylorus and in ileum at about 5 cm to the valva ileocecalis were made to carry out intestinal dialysis. The results showed that the clearance of the urea in treated groups with mannitol, regitine, and oxystarch was higher than that in the control group with normal saline solution(P < 0.05). The clearance of urea and creatinine in treated groups with atropine was higher than that in the control group(P < 0.05). There was no significant difference in the clearance of urea and creatinine among treated groups with different intestine pH value. PMID- 12080709 TI - [A model of closed epidural balloon inflation brain injury in rats]. AB - OBJECTIVES: To establish an animal model of closed brain epidural occupancy trauma in rats. METHODS: Sixty-eight Wistar rats were randomly divided into 7 groups: normal control group(Group A), sham-operated group(Group B), and 5 experimental groups. The last ones included: Group C in which water was injected 0.05 ml into balloon and kept for 1 hour; Group D injected 0.05 ml and kept for 3 hours; Group E injected 0.1 ml and kept for 1 hour; Group F injected 0.1 ml and kept for 2 hours. Group B, C, D were kept maintaining and observing for two days after operation, respectively. Group E was divided into EI (maintaining for two days) and EII (for three days). RESULTS: Of all the experimental groups, the mortality rate during acute phase was 100% in Group F, 28% in Group E, 50% in Group D, and 0% in Group C. Four rats died during the raising-up period in Group E. Both ICP and cerebral water content were increased in 46 survived rats, and nerve functions were damaged in all the experimental groups. Pathologic changes read as follows: Hemorrhage was found in damage-area; injury and dead cells were seen in compressed region. These changes became worse with time prolonged and injury degree developed. CONCLUSIONS: The method is simple, reproducible and economical, well showing the pathological classification of epidural occupancy trauma. PMID- 12080710 TI - [Effects of Ganoderma lucidum (Leyss ex Fr) Karst compound on the proliferation and differentiation of K562 leukemic cells]. AB - A series of experiments including cell culture and benzidine staining test were undertaken to investigate the effects of Ganoderma lucidum(Leyss ex Fr) Karst Compound(GLC) on the proliferation and differentiation of K562 leukemic cells. The results showed that different concentrations of GLC(from 4 mg.ml-1 to 12 mg.ml-1) could promote human bone marrow granulocyte-macrophage colony forming unit (CFU-GM) proliferation, but suppressed the growth of K562 leukemic cell colonies, and IC50 was 9.2 mg.ml-1. The data from liquid culture demonstrated that GLC could suppress K562 cells proliferation in a dose-dependent(from 4 mg.ml 1 to 20 mg.ml-1) and time-dependent(from 1-5 days) manner. K562 cells could be induced to differentiate into more mature erythrocytic cells by 4 mg.ml-1 and 8 mg.ml-1 GLC. It is concluded that GLC may be a good medicine for leukemia therapy. PMID- 12080711 TI - [Experimental studies on hyperthermic distilled water combining with saline and dextran for preventing intraperitoneal cancer recurrence due to neoplasm seeding]. AB - In order to know the efficacy of hyperthermic distilled water combining with saline and dextran (DSD) to prevent intraperitoneal cancer from recurrence due to neoplasm seeding, we carried out the experimental studies, including an observation on inhibiting cancer cells from implantation in vivo by abdominal cavity lavation, cytotoxic test and attachment test in vitro. The results of the tests in vitro showed no viable tumor cell exposed to 45 degrees C distilled water for 20 minutes in the tissue culture wells, meanwhile, dextran-40 partially blocked attachment of B16-F10 cells to gelatinized wells. In vivo test, DSD effectively prevented neoplasm seeding of B16-F10 viable melanoma cells in C57BL/6j mouse abdominal cavity and had no obvious toxicity. The efficiency was 70%; when cisplatin was added, the efficiency was enhanced to 88.9% with slight toxicity. These data indicate that the local application of hyperthermic distilled water combined with saline and DSD may prevent intraperitoneal neoplasm from recurring. PMID- 12080712 TI - [Mutation and expression of p16 in human ovarian neoplasms]. AB - OBJECTIVES: To detect mutation and expression of p16 in human ovarian neoplasms and to understand the relations between p16 and development of ovarian neoplasms. METHODS: Polymerase chain reaction-single strand polymorphism analysis and immunohistochemistry were respectively used to detect mutations in exon 1 and 2 and protein expression in 14 patients with ovarian neoplasms. Metastasis tissue of 5 mentioned patients were screened at the same time. RESULTS: No expression of p16 was found in 7 ovarian neoplasm tissues. No mutation in exon 1 of p16 was found in any patient tissues, but mutations in exon 2 were found in 4 patients tissues and expression of p16 was not found in 2 of them. There was correspondence between primary focus and metastasis in either gene mutation or protein expression. CONCLUSIONS: Inactivation of p16 may be involved in the development of ovarian neoplasms. Gene mutation is a kind of inactivation. Alterations of p16 probably occur before metastasis. PMID- 12080713 TI - [Effect of benazepril on renal interstitial inflammatory cell infiltration in rats with unilateral ureteral obstruction]. AB - Effect of benazepril on renal interstitial inflammatory cell infiltration was observed in rats with unilateral ureteral obstruction(UUO). The results showed that the relative volume of interstitium and number of infiltrating inflammatory cell on the fifth day were significantly less in rats with benazepril than the control group, but the difference between two groups lose significance on the tenth day. These data indicate that benazepril can ameliorate early phase of renal interstitial inflammation in UUO. PMID- 12080714 TI - [Apoptosis of CD4(+) T cells during experimental autoimmune neuritis in Wistar rats]. AB - The present study describes apoptosis of T cells in the sciatic nerve in Wistar rats with experimental autoimmune neuritis(EAN). Morphological characterizations of apoptosis were found at the 18th day from onset, peaked at the 22nd day by immunocytochemical analysis. In situ end labeling(ISEL) techniques confirmed the presence of DNA fragmentation in CD4(+) T cells. It is suggested that apoptosis is an important clearance mechanism of infiltrated T cells in the peripheral nervous system(PNS) in EAN. PMID- 12080715 TI - [Experimental and clinical study to remove lead and to prevent lead damage of kidney with salt of high dose KI]. AB - OBJECTIVE: To study and make a salt prevent lead poisoning. METHODS: SD rats were given 0.25% lead acetate by food intake and were given 10 mg.kg-1.d-1, 20 mg.kg 1.d-1 and 30 mg.kg-1.d-1 potassium iodide from drinking water in respective groups, in order to look for the dose of KI which prevents lead poisoning. The salt to prevent lead poisoning was made, 50 workers were given the salt as to flavouring of daily food intake. PbB, Pbu, ZPP, delta-ALA, NAG(N-acetyl-beta-D glucosaminidase), RP(Retinol protein) were observed. RESULTS: The dose of KI to prevent experimental lead poisoning of SD rats was 10 mg.kg-1.d-1 by oral intake. People who contacted lead had an increase of urine lead in the third day, and had most urine lead on ninth day after taking the salt with KI. Normal urine lead on the fourth week. Pre- and post-oral intake of the salt with high dose KI, Hb, ZPP, delta-ALA, PbB, NAG and Rp the people mentioned above had the following indexes, (101.2 and 128) g.L-1 (P = 0.05), (33.5 and 7.7) micrograms.g-1 Hb(P < 0.01), (123.12 and 65.35) mumol.L-1 (P < 0.05), (2.123 and 0.85) mumol.L-1 (P < 0.01), (20.9 and 6.7) U.L-1 (P < 0.01), (1.5 and 0.34) mg.L-1 (P < 0.05) respectively. CONCLUSION: The salt with 1% KI may remove lead and prevent lead damage of the kidney. PMID- 12080716 TI - [Relationship between nitric oxide and oxygen free radicals in different duration of diabetes in rat kidney]. AB - To study the relationship between nitric oxide(NO) and oxygen free radicals in rat kidney of diabetes mellitus(DM), the authors examined the changes of renal tissue NO level and nitric oxide synthase(NOS) activity, lipid peroxidation(LPO) level and superoxide dismutase(SOD), peroxidase(POD) and catalase (CAT) activities in streptozotocin(STZ)-induced diabetic rats and normal controls(NC) at the 2nd, 8th- and 16th-week. The results showed that renal-tissue NO level and NOS activity were increased at the 2nd week, but decreased at the 8th- and 16th week; LPO levels were significantly elevated in all DM groups than those in NC groups. SOD, POD and CAT activities decreased significantly in the 8th and 16th week DM groups compared with those in NC groups. The results suggest that NO level in kidney of STZ-induced diabetic rats dynamically change with the disease course of development. This change may be related with the change of anti oxidation in kidney which high glucose induces. PMID- 12080717 TI - [Protection of naoyi-an granule in ischemic brain injury secondary to intracerebral hemorrhage in rats]. AB - AIM: To explore the protective effect and its mechanism of the traditional Chinese medicine complex, naoyi-an granule(NYAG), on intracerebral hemorrhage(ICH) in rats. METHODS: Collagenase-induced ICH rats were used. Inducible nitric oxide synthase(iNOS) activity of ischemic cerebral cortex surrounding the intracerebral hematoma was assayed by monitoring the conversion of L-3H-arginine to L-3H-citrulline. Pathologic changes in the same area were observed with transmission electron microscope on the 2nd day, the 4th day and the 7th day after ICH. RESULTS: The iNOS activities of NYA-treated group decreased remarkably(P < 0.01) compared with those of the saline-treated group. Ultrastructural observation also indicated that NYAG ameliorated the brain edema, cell degeneration and necrosis. CONCLUSIONS: The inhibitory effect of NYAG on iNOS activity may be related to its protection against ischemic brain injury secondary to ICH. PMID- 12080718 TI - [Clinical analysis on 50 cases of heat invasion of blood chamber in the traditional Chinese medicine treatment according to differentiation]. AB - According to the clinical picture of 50 patients with invasion of blood chamber by heat, the patients were diagnosed for menstruation just coming type or just disappearing type or internal heat in yangming type in invasion of blood chamber by heat. They were treated with drugs to soothe and regulate the liver. The result that 16 cases(32%) were cured, 22 cases(20%) effective and 2 cases(4%) ineffective. The total effective rates were 96%. PMID- 12080719 TI - [Fluid therapy for acute brain edema in children]. PMID- 12080720 TI - [Application of intracoronary ultrasound in directional coronary atherectomy]. AB - Intracoronary ultrasound(ICUS) was successfully applied in 32 patients who accepted directional coronary atherectomy(DCA). After the procedure, the mean internal diameter of coronary artery increased from 1.06 mm to 3.59 mm, and the residual stenosis was < 25%. No severe complications, such as rupture of perforation, occurred. The clinical application of ICUS in DCA may improve the therapeutic effects as well as success rate of DCA, and may minimize the incidence of complications. PMID- 12080721 TI - [Chemoembolization of primary hepatic carcinoma with portal vein tumor thrombus]. AB - Forty cases of primary hepatic carcinoma(PHC) with portal vein tumor thrombus(PTT) underwent chemoembolization twice or more. The total effective rate was 67.5%. The survival rates of half, one, and three years were 75.0%, 12.5%, and 2.5% respectively after the treatment. The mean survival time was 9 months. The effective rate of the cases with the branch PTT was higher than other cases(P < 0.05). The influence caused by the average treatment interval had no significant difference(P > 0.05). It is suggested that (1) in case of controlling the applications, TAI + TAE is an effective therapy of PHC with PTT; (2) no matter where PTT is, an active management must be adopted; (3) the average treatment interval shall be decided according to conditions of the patients. PMID- 12080722 TI - [Relationship between the effect of carbamazepine on SCE frequencies and folic acid in epileptic patients]. AB - Peripheral blood lymphocyte sister chromatid exchange(SCE) frequencies, serum folic acid(FA) levels were examined in 15 epileptic patients treated with carbamazepine(CBZ), and another 15 epileptic patients treated with CBZ and folic acid(FA). The untreated epileptic patients and the healthy subjects served as control. The results showed that SCE frequencies were significantly higher in CBZ Group, compared with CBZ plus FA Group and control(P < 0.01). Serum FA levels were lower in CBZ Group compared with healthy control(P < 0.01). It suggests that CBZ can induce the increase of SCE frequencies. Supplementation with FA may effectively prevent chromosome DNA damage induced by CBZ. PMID- 12080723 TI - [Changes of left ventricular diastolic function in hypertensive patients with left atrial enlargement]. AB - In order to find out the relationship between the left atrial enlargement and left ventricular diastolic function in hypertension, radionuclide ventriculography was performed in 45 hypertensive patients with normal left ventricular mass index(LVMI), including 23 with normal left atrium(Group I), 22 with left atrial enlargement(Group II), and 28 matched subjects as control(Group III). The results showed that the left ventricular diastolic function in Group I was normal, but abnormal in Group II. It is considered that left atrial enlargement and left ventricular diastolic dysfunction are the early signs of the hypertensive heart disease. PMID- 12080724 TI - [Survey of jugular bulb with MRA in sudden sensorineural hearing loss]. AB - To evaluate the correlation between the size and position of the jugular bulb and the sudden sensorineural hearing loss(SSHL), the diameter and height of all jugular bulbs were measured with magnetic resonance angiography and analysed with statistics in 15 cases with SSHL and 35 cases without SSHL(as control). The ipsilateral and the contralateral bulbs of the SSHL were more asymmetrical than that of the control. The mean diameter and height of the ipsilateral bulbs of SSHL were larger than that of the dominant side bulbs of the control. It indicates that the heteroplasia of jugular bulbs which were greatly enlarged and bulgy upward might be related to SSHL. PMID- 12080725 TI - [Relationship between hypotension during hemodialysis and autonomic dysfunction in diabetic hemodialysis patients]. AB - OBJECTIVE: To observe the change of autonomic nerve function in diabetes mellitus(DM) patients who receive hemodialysis and to explore the relationship between this change and hypotension during hemodialysis. METHODS: Twenty-five DM patients and 40 chronic glomerulonephritis(CGN) patients who received hemodialysis were checked. We tested their coefficient of variation R-R(CVR-R) in ECG, difference of systolic blood pressure between lying and standing positions, and serum parathyroid hormone(PTH); monitored the decrease of blood pressure during hemodialysis. RESULTS: The rate(80.0%) of autonomic dysfunction(AD) in DM group is significantly higher than that(57.5%, P < 0.05) in CGN group. The rate(37.2%) of hypotension in those who had AD was significantly higher than that(18.2%, P < 0.05) in those who had normal autonomic nerve function during hemodialysis. No significant difference was found in PTH between each group. CONCLUSION: DM patients who receive hemodialysis are often complicated with AD, which is one of the reasons that causes hypotension during hemodialysis. PMID- 12080726 TI - [One hundred and twenty-four cases of diffuse pulmonary disease diagnosed by transbronchial lung biopsy via fiberbronchoscope]. AB - OBJECTIVE: To understand the diagnostic value in diffuse pulmonary disease examined with transbronchial lung biopsy(TBLB) via fiber bronchoscope. METHOD: One hundred and twenty-four cases of diffuse pulmonary disease were diagnosed with TBLB via fiberbronchoscope. RESULT: Confirmed diagnosis was obtained in 91 patients; the total diagnostic rate was 73.39%. The kinds of the diseases included lung cancer, lung tuberculosis, diffuse interstitial pulmonary fibrosis, pulmonary sarcoidosis, and pulmonary lesion in collageous diseases. There were no other serious complications except for bloody or blood-streaking sputum, and pneumothorax occurred occasionally. CONCLUSION: It is suggested that TBLB is an easy, safe and useful method for diagnosing diffuse pulmonary diseases. PMID- 12080727 TI - [Acute flare-up of senile cardiopulmonary disease and multi-system organ failure]. AB - In this paper we report 163 cases of senile cardiopulmonary disease. Among them, 60 sustained multi-system organ failure(36.9%). Of those 60, 19 patients died(death rate 31.7%). We believe that most of the acute attacks of senile cardiopulmonary disease were induced by pneumonia. Therefore it is very important to control infection while treating respiratory-circulation failure, pulmonary encephalopathia, shock, acid-base imbalance, and other complications. PMID- 12080728 TI - [Indirect determination of berberine hydrochloride by atomic absorption spectrometry]. AB - A method for the indirect determination of berberine in tabellae berberine hydrochloride by ion pair extraction and atomic absorption spectrometry was described. Berberine reacted with inorganic complex tetrathiocyanate cobalt(II) to form a stable ion pair associated complex at certain pH value. The ion pair associated complex was extracted with organic solvent and then transferred into aqueous solution. The content of cobalt in the solution was measured by flame atomic absorption spectrometry and the amount of berberine hydrochloride was calculated according to the content of cobalt. The result of the present method agreed with pharmacopeia method. This method is sensitive, dependable, precise, and applicable. PMID- 12080729 TI - [Sample preparation for electron microscope with tissues fixed with formalin and embedded in paraffin]. AB - A method of sample preparation for electron microscope is introduced. After paraffin-embedded tissues, fresh and fixed on time, were deparaffined, the tissue were fixed in 4% glutaraldehyde for 24 hours, in 1% osmic acid for 1-2 hours, and dehydrated with acetone and embedded in Epon812; then they were cut into ultrathin sections. By this method, better ultrastructure of paraffin-embedded tissues was obtained, and the method could satisfy the demand for observation under electron microscope. In addition, some important proceedings were also discussed in the paper. PMID- 12080731 TI - [8 cases of adrenal gland cysts]. PMID- 12080730 TI - [Momordica charantia proteins against coxsackievirus B3 infection in vitro]. PMID- 12080732 TI - [Relationship between the expression of bcl-2 and proliferation cell nuclear antigen in islet cells and prevention of complete Freund's adjuvant on insulitis and diabetes in nonobese diabetic mice]. PMID- 12080733 TI - [Relationship between T cell subgroups and HBV markers in the patients with chronic hepatitis B]. PMID- 12080734 TI - [Effect of hyperbaric oxygenation on cortisol and T lymphocyte subsets in patients with brain injuries]. PMID- 12080735 TI - [Effect of valociclovir hydrochloride on patients with herpes zoster]. PMID- 12080736 TI - [A novel method for rotated images registration based on wavelet transform]. AB - This paper presents a novel approach to image registration which addresses the problem of rotation correction. METHOD: 1. 2-DWT of medical images. 2. Maxima of wavelet coefficients (MWC) were used as feature points. 3. Starting from last level of decomposition and then iteratively for each level going up, find the best match between MWC of input image and rotated MWC of reference image. The best rotation is refined iteratively and with increasing accuracy. RESULTS: The performance of the method was compared to that of the traditional methods and found it to be superior. CONCLUSIONS: Images can be fast registrated with high accuracy and less sensitivity to noise. PMID- 12080737 TI - [Interactive 2D/3D image processing, registration and display system]. AB - This paper presents an integrated system for interactive 2D/3D image processing, registration and display. The main image processing algorithm in the system is image filtering, enhancement, adaptive B-snake model, image registration, pseudo color representation and 3D volume rendering. The system provides good visulization interface and image tool. PMID- 12080739 TI - [Segmentation of osteomas X-ray image using mathematical morphology]. AB - In this paper, we introduce a method for the segmentation of osteomas X-ray images. This method is mainly based on mathematical morphology. Structuring elements are selected according to the shape of osteomas. After gray scale morphology operations, osteomas boundary is extracted by histogram threshold. Further processing of resulting boundary with binary morphological operation produces a closed osteomas contour. So osteomas is isolated from background. PMID- 12080738 TI - [The study of multi-phrase multi-criterion steretactic radiotherapy treatment planning]. AB - Steretactic radiotherapy treatment planning is an important part of X-knife radiosurgery. In this paper the theory of multi-phrase-multi-criterion steretactic radiotherapy treatment planning is proposed, which is composed of three phrases. In the first stage, linear programming is used to reduce the resolution space. A kind of quadriatic programming is used in the second phrase in order to get high and well-distributed dose distribution. In the third phrase, an improved method of constrained simulated annealing is used to get the effect of conforming with the clinical practice. In correspondence with three-phrases optimization, three criterions are used respectively. The experiment results show that the theory proposed is correct. PMID- 12080740 TI - [The study of architecture of portable multiparameter physiological monitor]. AB - The portable multiparameter physiological monitor now becomes a hot point of competition of international manufactures. The paper uses the UPM-2001 patient monitor as an example, studies the key points of design of portable-type monitor which breaks through the traditional architecture of desk-type monitor. PMID- 12080741 TI - [Development of the IMP900 ECG & VT patient monitoring system]. AB - A new respiration monitor was developed with this paper, which can monitor tidal volume continuously based on the impedance technique as well ad ECG at the same time. Advanced digital storage display technique and modular hardware design are adopted in this system. One by one calibration method was introduced to solve problem of individual differences in the measurement of tidal volume, which make it viable to monitor Tidal volume with impedance technique. The system possesses characteristic of high ratio of performance to cost. Clinical tests of the system have been made for nonsurgical patients, the results show that the system meets the basic clinical requirements, and its wide clinical applications are expected in the future. PMID- 12080742 TI - [Analysis of oscillatory potentials of flectroretinogram]. AB - This study presents a method for the analysis of oscillatory potentials (OPs) of electroretinogram (ERG) in frequency domain and time domain. OPs from 9 cases (9 eyes) of branch retinal vein occlusion, 9 cases (9 eyes) of central retinal vein occlusion and 40 normal control eyes were tested. The frequency spectra of the OPs were derived through Fast Fourier Transform. The mean frequency spectra, the total power, the maximum magnitude, the maximum power and the dominant frequency in frequency domain of the OPs were calculated. The OPs waveforms were re constructed with the OPs frequency spectra data by counter-Fast Fourier Transform. In time domain, the latencies, the amplitudes and the sum of the amplitudes of the four peaks of OPs were measured. PMID- 12080743 TI - [Study on a new type of endoscope based on micro motor]. AB - The application of Micro Electro Mechanical System (MEMS) on the fields of biomedical engineering is a newly technological field. Applying a micro motor, whose diameter is only 2 mm, as driving device of this system, this paper presented a new type of medical endoscope whose front tip of diagnosing tube could work automatically. PMID- 12080744 TI - [The development of a new laryngoscope]. AB - A new type of laryngoscope, with powerful cold beam and fiber optic bundle, has been developed. It is a reliable aid of intubates during the field of emergency treatment and the foging anaesthesia can be done at the same time. The waterproof handle can be immersed in special liquids for thorough, effective cleaning and sterilization. PMID- 12080745 TI - [An ideal imaging quality can be obtained from the evenly scattering x-ray density]. AB - Since X-ray image quality was evaluated by the self-delivering function, the limitations in the application of fixative focus X-ray tube seem more outstanding. How to improve the quality of the x-ray image has been one of the focus problems in the field of the x-ray diagnosis. If we use the x-ray tube of the evenly scattering x-ray density as the radio source, we can get the ideal images. PMID- 12080746 TI - [Progress in the technology of medical ultrasonography equipments and current status of the market]. PMID- 12080747 TI - [Determination of the cutaneous viability of skin following incomplete avulsion and its treatment]. AB - OBJECTIVE: In the determination of the viability of skin following incomplete avulsion, subjective criteria such as color, skin temperature, pressure reaction and the stab bleeding would often give a high rate of failure. In order to resolve this problem, a retrospective study was carried out. METHODS: In 27 patients, there were 18 males and 9 females, the age ranged from 7 to 41 years old. In operation, the blood supply of the skin was determined by above subjective criteria carefully. RESULTS: After operation, 5 cases had total survival, 7 cases had peripheral or small area necrosis and 15 cases resulted in large area of necrosis. CONCLUSION: The conclusion was that if the incompletely avulsed skin showed sign of being compressed and squeezed, or the incompletely avulsed skin had uncertain or unstable circulatory status, even though the circulatory status being good, active attitude should be given to debridement in complete removal of the avulsed skin in order to improve the successful rate. For the other 2 cases with degloving injuries of large area of the limbs, the avulsed skin was made into a subdermal vascular network skin flap and several axial incisions were made to save the blood circulation of flap. The result was satisfactory and the vital tissues were preserved and used to the greatest extent. PMID- 12080748 TI - [Ilio-inguinal-hypogastric vascular network skin flap with common pedicle in the treatment of degloving injury of multiple fingers]. AB - OBJECTIVE: In order to solve the difficult problem of one-stage repair of degloving injury of multiple fingers, the common pedicled ilio-inguinal hypogastric subdermal vascular network skin flap was designed and the multi-lobes skin flap was performed subsequently. METHODS: From 1993 to 1996, there were 5 cases with degloving injuries of multiple fingers were treated by this flap. There were 2 males and 3 females and the age ranged from 7 to 19 years old. RESULTS: After operation, the pedicles of the flap was detached between 12 to 16 days and all of the flaps survived completely. Patients were followed up for 6-18 months. After repair, the contour and skin colour of the digits were excellent, and the motion of the interphalangeal joints and skin sensation were good. CONCLUSION: The conclusion was as follows: The newly designed skin flap was characterized by the advantages of duration of treatment being short, excellent contour and more rapid recovery of function. It could be used for one-stage repair of degloving injury of multiple fingers. PMID- 12080749 TI - [Reinnervation of skin flap by end-to-side neuro-anastomosis in the repair of soft tissue defect of heel]. AB - OBJECTIVE: The repair of soft tissue defect of heel by traditional operation did not restore the sensation of the heel. METHODS: In order to solve this problem, the saphenous neurovascular skin flap reinnervated by end-to-side neuro anastomosis was designed and 3 male patients with soft tissue defects of the heel were so treated. Grossgraft of saphenous neurovascular skin flap was employed for repairing the soft tissue defects of heel and the pedicle was divided at 21 days after operation. The end-to-side neuro-anastomosis was used to reinnervate not only the skim flap, but also the skin area of the medial malleolus, medial aspect of the foot and the big toe of the donor limb. RESULTS: After follow-up of 6-12 months, the walking and weight bearing functions of the affected limbs were good, the contour of the grafted area was satisfactory, and the recovery of sensation of the skin flap, the medial malleolus, the medial aspect of the foot and the big toe was observed. CONCLUSION: 1. Crossgraft of the saphenous skin flap was an effective method to repair the soft tissue defect of the heel; 2. End-to-side neuro-anastomosis could restore the sensation of the skin flap. PMID- 12080750 TI - [Repair of soft tissue defect of heel with myocutaneous flap of flexor digitorum brevis]. AB - OBJECTIVE: To investigate the repairing method of soft tissue defect of heel, pedicled island myocutaneous flap of flexor digitorum brevis was designed. METHODS: From 1984 to 1997, 26 cases with soft tissue defect of heel were adopted in the clinical trial. Among them, the were 18 males, 8 females and the age ranged from 15 to 60 years old. The area of wound ranged from 2.5 cm x 1.5 cm to 8.0 cm x 6.0 cm. RESULTS: After operation, all of the flaps survived. They were followed up for 9 to 72 months. All of the flaps had primary healing except in one there was infection of peripheral of the flap. The contour of heel was satisfactory the sensation of flap was good and the weight-bearing function was also successful. CONCLUSION: It was concluded that the myocutaneous flap of flexor digitorum brevis could be used to repair the soft tissue defect of heel because of its nearby position, hidden location, good recovery of skin sensation and weight-bearing function, Besides, the procedure of this operation was simple and the anti-inflammatory potential of the flap was high. However, Because of the limited donor area, the pre-operative design was important. PMID- 12080751 TI - [Reconstruction and repair after resection of soft tissue sarcoma]. AB - OBJECTIVE: To emphasize the importance of reconstruction and repair after resection of soft tissue sarcoma. METHODS: From November 1990 to November 1996, in 107 cases of soft tissue sarcoma 32 cases had received various reconstructive or reparative operations. Among the 32 cases, 4 cases were primary sarcomas and 28 cases were recurrent sarcomas. In surgical grading, 3 cases were of I B, 3 cases of II A and 26 cases of II B. Radical resection was performed in 13 cases, widen local resection in 17 cases and local excision in 2 cases. After operation, 13 cases received chemotherapy and 7 cases received radiotherapy. Reconstruction of blood vessels was performed in 3 cases, reconstruction of kinetic function in 16 cases, and repair of defect was carried out in 23 cases. RESULTS: Thirty patients were followed up for 4 months to 6 years and 6 months. The clinical results showed that the local control rate of sarcoma was 80%, limb-salvage rate after reconstruction of blood vessels was 100%, the excellent and good rate after reconstruction of kinetic function was 87.5%, and the survival rate of the tissue flap of transplantation and transposition was 96%. CONCLUSION: It was concluded that the reconstruction and repair after resection of soft tissue sarcoma was the extension of operative treatment, and was very important in lowering the recurrence rate and improving the life quality of the sufferings. PMID- 12080752 TI - [A new method for repair of huge incisional hernia of abdominal wall]. AB - OBJECTIVE: To repair the huge incisional hernia of abdominal wall, a new surgical method was introduced. METHODS: Eight cases of huge ventral incisional hernia, developed in 3 months to 12 months after operation, were treated in this new method with the defects ranged from 8 cm x 4 cm to 12 cm x 6 cm. RESULTS: They were followed up for 6 months to 18 months after operation. The clinical results showed that all of the 8 cases recovered satisfactorily without recurrence. CONCLUSION: The new method was recommendable for its advantages of easier manipulation, shortened time, no tissue reaction and less tissue trauma from operation. PMID- 12080753 TI - [Axial aponeurotic flap combined with skin graft in the treatment of lower lid ectropion and severe infra-orbital soft tissue depression]. AB - OBJECTIVE: The correction of ectropion of lower lid and severe infra-orbital soft tissue depression is very difficult. Former methods included simple skin graft, tubed graft, transfer of local skin flap and so on. These methods had some disadvantages, such as not enough tissue to fill the depression, too much damage done to the donor area and operation in stages required. METHODS: After investigation on the anatomy of temporal region, designed the following method. Combined transfer of the galea aponeurotica and temporal fascia was used to repair severe infra-orbital soft tissue depression and ectropion of lower lid in 6 cases. RESULTS: It was discovered that the combined transfer of the galea aponeurotica and temporal fascia was rich in blood circulation because they received blood supply from parietal branch of superficial temporal artery and could be transferred to a distance as far as 15-18 cm. The skin graft used to cover the fascia usually resulted in good survival. This technique was used in 6 cases with good success. CONCLUSION: This method had some merits such as the tissue flap had good blood supply, little damage done to the donor area, good correction of the severe depression, good appearance following correction, operation done under local anesthesia and completed in one-stage operation. PMID- 12080754 TI - [The clinical application of scalp expansion in the repair of cicatricial baldness in children]. AB - OBJECTIVE: To introduce the clinical application of the expanded graft from scalp in the repair of cicatricial baldness in children. METHODS: 45 cases with baldness following burn from 1988 to 1998 were reported. All of these patients (age ranged from 5-11 years) were treated by soft tissue expander. RESULTS: 5 cases were followed up for 1-2 years, the clinical results showed that the result from the graft of scalp expansion was satisfactory, and the long-term follow-up revealed that the hair in expended area and that in normal area was almost the same except the orientation of hair distribution had some difference. CONCLUSION: The head scalp expansion might be the first choice in the repair of cicatricial baldness following burn. PMID- 12080755 TI - [Interaction of cell adhesion to materials in tissue engineering]. AB - OBJECTIVE: To investigate the adhesive interactions of cells with materials and the effects of material properties on cell adhesion in tissue engineering. METHODS: By looking up the recent literatures dealt with adhesive interactions of cells with materials and reviewing previous work on the adhesion of tissue derived cells to materials. RESULTS: The adhesion characteristics of cells to materials not only depend on the nature of materials, including bulk and surface properties, surface modification, surface morphology, net charge, porosity and degradation rate, but also on the expression of cell surface molecules and their interaction with the material. CONCLUSION: The quantitative measure and biophysical mechanisms of cell adhesion to materials might be very important in tissue engineering. PMID- 12080756 TI - [Ceramic-like xenogeneic bone combining with bone marrow]. AB - OBJECTIVE: This experimental study was aim to investigate the osteogenesis of ceramic-like xenogeneic bone (CXB) combining with bone marrow (BM). METHODS: The CXB combining BM was implanted into the sacrospinalis muscle of rabbits, and CXB implanted alone was used as control. Eighteen Japanese rabbits with long ear were used. The size of CXB was 5 mm x 5 mm x 5 mm, and the implanted materials were taken out at 2, 4, 8, 12, 16 and 24 weeks after implantation. The histological and histochemical characteristics were investigated. RESULTS: There existed cartilage and new bone in the groups of CXB combining BM in 2 weeks. Later, be cartilage turned out to the bone and in eight weeks the medullary cavity appeared. However, as the time went on, new bone formation increased and typical osteogenesis could be found. While in the groups of CXB alone, no formation of new bone or cartilage was found. CONCLUSION: The implantation of CXB combined with BM could result in new bone formation in the way of osteoconduation, osteoinduction, and providing, osteoblasts or chondroblasts. It could be an ideal bone substitute, and its clinical use in future seemed very hopeful. PMID- 12080757 TI - [Analysis of the causes of necrosis of retained flap on tibial side after transplantation of the great toe nail flap]. AB - OBJECTIVE: In order to investigate the causes of necrosis of the retained flap after transplantation of the great nail flap, according to the principle of plastic surgery, the following operation was designed. METHODS: A tongue-shaped great toe flap was made with its base on the tibial side of the great toe and the width of the base was 1 cm to 1.5 cm. The flap contained the supporting vessels and nerves and the periosteum was also preserved. RESULTS: From June 1982 to November 1997, the flap was used to repair 267 case, 277 fingers defects. After operation, the retained flap on the tibial side developed necrosis in 13 sides, an incidence of 4.69%, and in 78 sides, there was necrosis on the grafted skin on the donor site, an incidence of 28.7%. Most of the wounds healed after changing dressing, while 37 wounds healed from another skin graft. Of these cases, 178 cases had been followed up for 5 months to 16 years. It was found that the retained tibial tongue-shaped flap showed hyperplasia which extended to the weightbearing area of metatarso-phalangeal of great toe. It showed that the taking off of great toe nail flap according to first dorsal metatarsal artery (FDMA) I, II or III type, the incidence of development of necrosis of retained flap on tibial side showed no significant difference (P > 0.05). CONCLUSION: It was believed that the necrosis of the retained flap was not related to the FDMA classification but had relation with the design, operative technique and presence of infection. PMID- 12080758 TI - [Preliminary evaluation of xenotransplantation and utilization of native pig breeds in China]. AB - OBJECTIVE: To evaluate the general situation of the native pig breeds and the relative present study conditions for better development of xenotransplantation in China. METHODS: By comparing the profits between the pigs and the non-human primates as the potential donors for xenotransplantation and emphasizing the source of the pigs as the organ donors from the transplantation studies, we analyzed the possible values of the native pig breeds as the donors. RESULTS: As one of the richest resources in the world, the species variation and relative genetic stability of Chinese native pig breeds could be the very valuable resources for xenotransplantation study and utilization. As a reverse, the xenotransplantation could provide opportunity for more economically and environmentally utilization of the pigs besides as the meat supply. CONCLUSION: As a very valuable and potential resource of organ donor, the native pig breeds of China might be noticed by the xenotransplantation colleagues in the world. It is necessary, to keep the balance among the risk vs. benefit and the protection vs. utilization of this valuable resource. PMID- 12080759 TI - [Influence of modulatory activity of wound fluid on the characteristics of fibroblasts from irradiation and the action of phenytoin sodium]. AB - OBJECTIVE: Influence of irradiation and phenytoin sodium on modulatory activities of wound fluid on proliferation of fibroblasts and collagen synthesis was studied. METHODS: The male Wistar rats were used in this study. The rats were divided into irradiated and non-irradiated groups, and in each of them it was subdivided into phenytoin group and control. A 7 cm long incisional wound was made on the back of each rat, in which a polyvinyl alcohol sponge (PVAS) with a size of 1.0 cm x 0.4 cm was implanted into the wound and the wound was sutured up. The PVAS was prepared by rinsing in running water over night and then was boiled for 30 minutes. Before implantation, the sponge was immersed in phenytoin sodium solution (10 mg/l ml) or normal saline (as control). From each wound the wound fluid and fibroblasts were collected. The methods of incorporation of 3H were adopted to assess the proliferation of fibroblasts and synthesis of collagen. RESULTS: It was shown that proliferation of fibroblasts and collagen synthesis were stimulated by wound fluid remarkably on 5 to 8 days after wounding, and that 6 Gy to total-body irradiation wound decrease this effect. It was also noted that topical phenytoin sodium increased the modulatory activity of wound fluid irrespective of being irradiated or not. CONCLUSION: It could be drawn that, after total-body irradiation, stimulation of hyperplasia of fibroblasts and collagen synthesis by wound fluid was markedly lowered indicating the total-body irradiation resulted in changes of local conditions of the wound which was unbenefitted to repair of tissue cells, while phenytoin sodium could enhance the stimulating action of wound fluid on proliferation of fibroblasts and synthesis of collagen which was beneficial to wound healing. PMID- 12080760 TI - [Acceleration of wound healing in diabetic rats by PDWHF and its relation with its activity to stimulate procollagen I (alpha 1) gene expression]. AB - OBJECTIVE: The effect of platelet-derived wound healing factor (PDWHF) on wound healing in diabetic rats was studied. METHODS: Forty-four male SD rats were randomly divided into 2 groups. Thirty-two rats of experimental group accepted intraperitoneal injection of alloxan (1.5 mg/10 g body weight). Within one or two days after injection, while the blood sugar of the rats was higher than 180 mg/dl, the animal model of diabetic rat should have been established. Then a dorsal incision was given to every rat. After the addition of PDWHF (the experimental group) or bovine albumin (the control group), the incision was sutured up. Seven, ten and fourteen days after operation, the breaking strength of the wound was measured. On another hand, specimen from the wound was taken for the culture of fibroblasts. When the cultured fibroblasts have been incubated with 10% PDWHF for 4, 8 and 12 hours, the procollagen I (alpha 1) mRNA levels were examined respectively, and compared with those of control. RESULTS: Significant difference in wound breaking strength had been observed between PDWHF treated incisions and the control on 7, 10 and 14 days after wounding (P < 0.01). Experiment in vitro demonstrated that the procollagen I (alpha 1) mRNA levels in wound fibroblasts incubated with 10% PDWHF for 4, 8 and 12 hours were 0.9, 3.7 and 2.2 folds higher than those in fibroblasts in control. CONCLUSION: It was suggested that direct stimulation of procollagen I (alpha 1) gene expression was one of the ways that PDWHF played its role in accelerating wound healing. PMID- 12080761 TI - [Ischemic reperfusion injury of peripheral nerves]. PMID- 12080762 TI - [Improved method of obtaining medial plantar cutaneous graft and its clinical application]. AB - OBJECTIVE: To introduce a skin flap containing the middle cutaneous branch of the medial plantar artery. METHODS: Microanatomic study was performed on 8 fresh cadaveric feet, the arteries were dissected and infused with methylene blue to observe their vascular distribution and the skin area supplied by the middle cutaneous branch. Furthermore, the clinical application was reported. A local pedicled flap containing the middle cutaneous branch was used to repair the soft tissue defects of the foot in 7 patients, and free cutaneous graft was used to repair the skin defects of the fingers in 6 patients. RESULTS: The results showed that the medical plantar artery gave off 3 cutaneous branches to supply the medial aspect of the foot, among which the middle branch was the largest one and anastomosed with the other two branches. The skin flaps used clinically were all survived completely. CONCLUSION: Medial plantar cutaneous graft had a reliable blood supply, and it's one of the best choice in repairing small to middle sized skin defects of the foot and the fingers. PMID- 12080763 TI - [The study of new bio-membrane-bred epidermal cell grafts]. AB - OBJECTIVE: The human epidermal cells were bred on a kind of bio-membrane, the bio brane, in engineering a kind of new epidermal substitute, the bio-membrane bred cell graft. METHODS: Fresh and frozen grafts of biomembrane bred epidermal cells were transplanted into the full-thickness wounds of nude mice and those received simple Bio-brane were served as control. The wounds of the two groups were observed daily and biopsy was taken on the 3, 5, 7, 10, 21 and 35 days respectively. RESULTS: Epidermal cells could be cultured in vitro on the bio membrane reaching the sub-saturated state of 60 to 70 percents. The bio-membrane after being grafted the epidermal cells continued to proliferate and differentiate to form a layer of new epidermis. There was no difference between the fresh and the frozen bio-membranes. CONCLUSION: Bio-membrane bred with epidermal cells could be a kind of ideal epidermal substitute. PMID- 12080764 TI - [Expression and distribution of human blood type antigens in Chinese Neijiang pig tissues]. AB - OBJECTIVE: The expression and distribution of human blood type A, B, O antigens in Chinese Neijiang porcine were investigated. METHODS: The anti-human blood type A, B, H monoclonal antibodies were used as the primary antibody, and the immunohistochemistry EnVision technique was used to detect the expression of A, B, H antigens in tissues from 10 pigs. RESULTS: The B and H antigens were not found in all 10 pigs' tissues investigated. While the A antigen was found in all above tissues except vascular endothelial cells. CONCLUSION: The A antigen did not exist on vascular endothelial cells, so it did not play a role as a hyperactive antigens in pig to human xenotransplantation. However, in other tissues it might play roles in acute cellular rejection or chronic rejection. The future study on its role in pig to human xenotransplantation was needed. The expression of human blood type antigens in Neijiang pigs was different from other pig strains reported, it was suggested that it was possible to find a pig strain that contained less xenoantigen. PMID- 12080765 TI - [Morphological changes and electrophysiological study of motor neuron of spinal cord following delayed repair of peripheral nerve injury]. AB - OBJECTIVE: Following the delayed repair of peripheral nerve injury, the cell number of anterior horn of the spinal cord and its ultrastructural changes, motorneuron and its electrophysiological changes were investigated. METHODS: In 16 rabbits the common peroneal nerves of both sides being transected one year later were divided into four groups randomly: the degeneration group and regeneration of 1, 3 and 5 months groups. Another 4 rabbits were used for control. All transected common peroneal nerves underwent epineural suture except for the degeneration group the electrophysiological examination was carried out at 1, 3 and 5 months postoperatively. Retrograde labelling of the anterior horn cells was demonstrated and the cells were observed under light and electronmicroscope. RESULTS: 1. The number of labelled anterior horn cell in the spinal cord was 45% of the normal population after denervation for one year (P < 0.01). The number of labelled cells increased steadily from 48% to 57% and 68% of normal values at 1, 3 and 5 months following delayed nerve repair (P < 0.01). 2. The ultrastructure of the anterior horn cells of the recover gradually after repair. 3. With the progress of regeneration the latency become shortened, the conduction velocity was increased, the amplitude of action potential was increased. CONCLUSION: Following delayed repair of injury of peripheral nerve, the morphology of anterior horn cells of spinal cord and electrophysiological display all revealed evidence of regeneration, thus the late repair of injury of peripheral nerve was valid. PMID- 12080766 TI - [Observation of microvascular casting of avulsion injured skin flap under electron microscope]. AB - OBJECTIVE: To investigate the mechanism of necrosis of avulsion injured skin flap. METHODS: Six swine were used. An avulsed skin flap and traditional island flap were made on each hind leg of the swine respectively, and the latter was used as control. Then the microvascular casting of each flap was made routinely immediately following the injury and 72 hours later, and the vascular casting were observed under electron microscopic scanning. RESULTS: There were three characteristic changes in the avulsed skin flap: tearing of blood vessels, formation of complete thrombosis and incomplete thrombosis. If these changes were excessive in the avulsed flap, then the necrosis was the result, otherwise, the flap survived. CONCLUSION: The necrosis of avulsed flap was related to the type and extent of the damage of microvascular endothelial structure. So the degree of endothelial damage was the keypoint in the viability of avulsed flap. PMID- 12080767 TI - [Effect of gamma-interferon on the fibroblasts in hypertrophic scars]. AB - OBJECTIVE: To study the influence and mechanism of gamma-IFN on fibroblasts in hypertrophic scars(HTS). METHODS: The cultured fibroblastic cells were isolated from the hypertrophic scars of 10 patients. The fibroblasts were divided into two groups, one group was treated with gamma-IFN (100 U/ml, 5 days) and the other without gamma-IFN as control. The proliferative activity in both groups was investigated and compared by blood cytometer, the proportion of myofibroblast (MFB) and the ratio of apoptosis were examined and analysed between two groups by flow cytometry using alpha-smooth muscle actin (alpha-SMA) as marker. RESULTS: The proliferative activity was downregulated with gamma-IFN. In gamma-IFN treated group, the differentiation of MFB were reduced and the decreasing ratio was 3.2% at the 2nd day and up to 10.5% at the 8th day, then it reduced gradually. The apoptosic ratio is 17.7% in gamma-IFN treated group, and is 10.9% in control group. The difference was statistically significant. CONCLUSION: gamma-IFN could downregulate the proliferation of fibroblasts, decrease the differentiation of MFB and induce the apoptosis. It has beneficial effect in the treatment of hypertrophic scars(HTS). PMID- 12080768 TI - [Experimental study of incompetency of deep vein of lower limb]. AB - OBJECTIVE: The purpose of this study was to find the best material for valvular wrapping operation of deep vein of lower limb and to study the diagnostic value of colored Ultrasonic-Doppler for valvular incompetency of the deep vein and the function of the popliteal valve. METHODS: Strips of autogenous saphenous vein, autogenous fascia lata and pieces of polytetrafluroethylene artificial vessel were used respectively as the wrapping material for narrowing the valve in 30 dogs. The results of three different wrapping material were obtained by colored Ultrasonic-Doppler and transpopliteal venography in 78 patients. The hemokinetics of the popliteal valve was examined in 20 normal persons. RESULTS: In the saphenous vein and fascia lata groups, diffuse fibrosis and marked narrowing of the femoral vein were found, while in the group of artificial vein graft, the graft was intact without prominent fibrosis and narrowing of the vein. In comparing with the result of venography, the accuracy of diagnosis by colored Ultrasonic-Doppler was 91.86%. The femoral and popliteal venous valves closed at the same time when holding the breath, but the popliteal venous valve opened more widely than the femoral venous valve when the calf muscles of the leg contracted. CONCLUSION: It was suggested that the graft was the best material for valvular wrapping operation, and colored Ultrasonic-Doppler was an important and non invasive method for the diagnosis of incompetency of deep vein. The popliteal venous valve was the important barrier for protection of the function of deep vein. PMID- 12080769 TI - [Comparative study of the late results between two types of operations in the treatment of old derangement of distal radio-ulnar joint]. AB - OBJECTIVE: To evaluate the clinical results of Darrach's procedure and Sauve Kapandji's procedure in the treatment of old derangement of distal radioulnar joint. METHODS: Twenty-three patients with old derangement of distal radioular joint were divided into two groups, in which 11 patients received resection of distal end of ulna by Darrach's procedure and 12 patients received pseudoarthrosis operation of the distal ulna by Sauve-Kapandji's procedure. RESULTS: Fifty-six months after Darrach's procedure, complete relief of pain was obtained in 6 cases (P < 0.01), the flexion-extension movement improved from 104 degrees to 125 degrees (P < 0.01), rotation movement of the forearm improved from 106 degrees to 128 degrees (P < 0.01) and grippig strength improved from 17 kg to 28 kg (P < 0.01). Fifty-five months after Sauve-Kapandji's procedure, complete relief of pain was obtained in 9 cases (P < 0.01), the movement flexion-extension improved from 108 degrees to 126 degrees (P < 0.01), rotation movement of the forearm improved from 101 degrees to 135 degrees (P < 0.01) and grippig strength improved from 17 kg to 35 kg (P < 0.01). CONCLUSION: 1. The two operation showed no difference in relief of the wrist pain, improvement of the movement of the wrist and that of the forearm; 2. Sauve-Kapandji's procedure was superior to Darrach's procedure in increasing gripping strength; and 3. So the Sauve Kapandji's procedure was superior to Darrach's procedure, especially in the long term result, in the treatment of old derangement of the distal radioulnar joint. PMID- 12080770 TI - [Resection of nerve of wrist combining replacement of lunate with bone cement prosthesis in treatment of Kienbock's disease]. AB - OBJECTIVE: There are various methods used to treat the Kienbock's disease with various outcomes. This study investigated the effect of combining acrylic lunate prosthesis replacement with resection of articular branches to wrist from anterior and dorsal interosseous nerve. METHODS: From January 1992 to January 1997, six cases were included. All of the patients were in stage III according to Lichtman's classification. Besides replacement of the acrylic lunate prosthesis, segment of the pure sensory branches from anterior and posterior interosseous nerves were resected. RESULTS: Follow-up for 10 to 54 months (an average of 32 months) the pain was relieved in all patients except one. The range of movement of wrists were as followings: The extension of the wrist was 20 to 35 degrees (an average of 26 degrees) and flexion was 20 to 35 degrees (an average of 32 degrees). There was no improvement in radial and ulnar deviation. CONCLUSION: Lunate prosthesis replacement combined with resection of articular branches to wrist resulted in a painless wrist and was a satisfactory method. PMID- 12080771 TI - [Electrophysiological criteria for selection of operative procedure in the treatment of late incomplete obstetrical palsy]. AB - OBJECTIVE: Using electrophysiological method to evaluate the severity of incomplete obstetric paralysis brachial plexus injuries, and establish an electrophysiological criteria for selection of operative procedure in the treatment of cases of late stage. METHODS: Neurolysis was performed in 16 patients, and during the operation, the neuroma at the upper trunk was discovered. The electrophysiological study was carried out before and during operation to evaluate the conducting function of neuroma. In the follow-up the operative result was analysed by Mallet test. RESULTS: The preoperative study showed that the compound muscular active potential (CMAP) amplitude of supraspinatus deltoid and biceps were decreased more than 60% in comparison with that of the healthy side. After external neurolysis of the neuroma, by stimulation of the proximal and distal ends of the neuroma the average decrease of CMAP amplitude of the above three muscles was 37.45% +/- 20.97%, 47.85% +/- 26.23%, 47.05% +/- 21.23%, respectively and no significant improvement was observed in the shoulder and elbow function in all of the 16 cases. CONCLUSION: When the preoperative electrophysiological study found that the CMAP amplitude decreased more than 60% in comparison with that of the healthy side, transposition of nerve or nerve grafting was indicated. PMID- 12080772 TI - [Selective ventral rhizotomy for the treatment of spastic cerebral palsy]. AB - OBJECTIVE: To understand the influence of selective ventral rhizotomy on the function of lower extremity and to investigate its effect in the treatment of spastic cerebral palsy. METHODS: Ten domestic dogs were used in this study. Under anesthesia, the lumbar spinal canal was reached through posterior approach. The ventral rami consisted of left L5, L6 and L7 spinal nerves was separated. The electromyographic changes of each branch of the rami was examined by electrical stimulation. 60 percents of the L6 ventral nerve root which was the major innervation of the gastrocnemius muscle in dogs, and 30 percents of the other two rami were culted off. The muscle tone, the coordinated movement and balanced capacity of the dogs were observed. After one month, the electrophysiological changes of the biceps femoris, semitendinous, gastrocnemius and anterior tibial muscles were examined and compared with that of the identical muscles of the contralateral limbs. RESULTS: Selective ventral rhizotomy was well correspondent to the innervated muscles. After operation, the coordinated movement and balanced capacity were good, but the muscle tone of the corresponding muscle was lowered. This technique was applied on 3 patients with satisfactory short-term results. CONCLUSION: For the treatment of spastic cerebral palsy, it is worth trying the selective ventral rhizotomy. PMID- 12080773 TI - [The result of splenopneumopexy on patients with portal hypertension in children]. AB - OBJECTIVE: The purpose of this study was to study the effect of splenopneumopexy for patients with portal hypertension in children. METHODS: From March 1993 to April 1998, splenopneumopexy was performed on six children with portal hypertension. Doppler ultrasound and radionuclide were used to demonstrate the portopulmonary shunt after operation. RESULTS: The bleeding from the esophageal varices was controlled and the esophageal varices were eliminated gradually. The symptoms pertaining to hypertension were disappeared. The patency of the shunt was maintained without the formation of thrombosis. No pulmonary complication was observed. CONCLUSION: The results indicated that splenopneumopexy was a safe and effective procedure for patients with portal hypertension in children. PMID- 12080774 TI - [Repair of vagina with segment of vascularized ileal graft following pelvic exenteration for carcinoma of rectum]. AB - OBJECTIVE: To introduce a method to repair the vagina following pelvic exenteration for carcinoma of rectum in which the posterior wall of the vagina and cervix of the uterus were often involved. METHODS: From 1990 to 1997 segment of the vascularized ileum was used to repair the vagina in 5 cases, and in 2 of which the whole vagina was repaired while in the other 3 cases only the posterior wall of the vagina was repaired. RESULTS: All of the patients had successful results after operation repair. CONCLUSION: Vascularized graft was an ideal material for the repair of vagina defect following pelvic exenteration for carcinoma of rectum, because this material was easily accessible, and its vascular pedicle was long enough for its transferring to the perineal region and the ileum had good blood supply which made healing easy. The vagina following repair had a thick posterior wall, good elasticity and very little scar tissue surrounded. PMID- 12080775 TI - [Study of concurrent reconstruction of posterior wall of vagina with pedicled muscular flap of uterus after resection of rectum carcinoma]. AB - OBJECTIVE: To investigate the reconstructional method of posterior wall of vagina after the resection of rectum carcinoma. METHODS: From August 1991 to March 1996, 10 patients with rectum carcinoma were adopted in this study, among them, there were 4 cases belong to B stage of Dukas, and 6 cases belong to C stage of Dukes. In operation, rectum carcinoma and posterior wall of vagina were resected, and concurrent reconstruction was finished by using pedicled muscular flap of uterus. RESULTS: The effect of operation were satisfactory except one case who was failed because of insufficient blood supply of the flap. Followed-up for 3-6 months, the posterior wall of vagina healed in 7 cases, the width of vagina was 2-3 fingers and the depth was 5-6 cm. CONCLUSION: Reconstruction of the posterior wall of vagina with pedicled muscular flap of uterus was available in clinic. For its simplicity and feasibility, it's suitable for the resection of rectum carcinoma in which the posterior wall of vagina was infiltrated. PMID- 12080776 TI - [Repair of huge skin defect on leg and foot with multiple pedicled blocking randomized fasciocutaneous flap]. AB - OBJECTIVE: In order to increase the survival area of pedicled fasciocutaneous flap, a multiple pedicled blocking randomized fasciocutaneous flap was designed. METHODS: From January 1991 to September 1998, this technique was used to repair 33 cases, including 27 males and 6 females and the ages ranged from 6 to 58 years. All of the patients were suffered from traffic accidents. In these cases, 22 cases had skin defects of legs and feet with bone, nerve and tendon exposed, 5 cases had osteomyelitis as well as internal fixaters exposed and the other 6 had deformity from scar. The size of the flap was 25.0 cm x 13.0 cm x 2.4 cm at its maximum and 6.0 cm x 3.5 cm x 1.5 cm at its minimum. Based on the traditional blocking flap, according to the severity of the wound and conditions of the neighboring tissues, a flap having 2 to 4 orthogonal pedicles with a width of 1.5 to 3.0 cm was designed. The medical-graded stainless steel sheet was implanted below the deep fascia, and after blocking for 3 to 6 days, the side pedicles were divided. 6 to 14 days later, one of the two remaining pedicles was divided and was transferred to repair the defect. RESULTS: 31 cases were followed up for 6 months to 5 years without any trouble of the joints. The flap had a good external appearance and was high pressure-resistant. CONCLUSION: The multiple pedicled blocking randomized fasciocutaneous flap increased the size of the flap and the length to width ratio. It had the following advantages: manage at will, high resistance to infection and a large survival area of flap. PMID- 12080777 TI - [The application of expanded polytetrafluoroethylene in the treatment of facial depression with facial nerve palsy]. AB - OBJECTIVE: In order to investigate the effect of expanded polytetrafluoroethylene (e-PT-FE) in repair of facial depression. METHODS: From February 1997 to July 1998. The e-PTFE was used as a filling and suspending material for facial depression and facial nerve palsy in 6 cases. They were followed up for 2-16 months. RESULTS: After filling of the facial depression, the deformed side of the face became symmetrical to that of contralateral side, and those following the suspending technique using this material the affected eyelids could close completely and the oral corners were symmetrical in its static status. CONCLUSION: The expanded PTFE was a safe, biocompatible and easily applied material, especially served as a filling or suspending material in the repair of facial depression with facial nerve palsy. PMID- 12080778 TI - [Culture of the transformed human embryonic tendon cells and its biological characteristics in vitro]. AB - OBJECTIVE: This paper was to study the biological characteristics of the transformed human embryonic tendon cells, the relation between cell growth and culture conditions, and to compare these features with that of human embryonic tendon cells. METHODS: The pts A58H plasmid had successfully used to transform a tendon cell line from human embryo in our past work. The human embryonic tendon cells and the transformed human embryonic tendon cells were cultured in vitro. In different culture conditions, the growth curve were drawn respectively. Population dependence and proliferation capability of the cells were investigated through plate cloning test and soft agar culture. The collagen secreted by cells was identified by immunohistochemical method. RESULTS: In routine culture condition, the growth properties of the human embryonic tendon cell and transformed cells were almost identical. The growth properties of the transformed cells were not changed when the cells were frozen storage. There were changes of growth characteristics of the transformed cells when the culture temperature was changed. The transformed cells could subcultured continually and permanently. The proliferation capability of the transformed cells were stronger than that of the human embryonic tendon cells. Moreover, the growth of the transformed cells was serum-dependent, and the phenomenon of contact inhibition was observed. The transformed cells were not able to grow on soft agar culture. They had the capacity of secreting collagen type I. CONCLUSION: The transformed human embryonic tendon cells could be subcultured continually and permanently, and their growth could be controlled by changing their culture conditions and they had no malignant tendency in biological characteristics. They could be taken as an ideal experimental material for tendon engineering. PMID- 12080779 TI - [The experimental study of membrane-guided tissue regeneration of bone]. AB - OBJECTIVE: To confirm membrane-guided tissue regeneration in the healing course of segmental bone defects and study the mechanism. METHODS: Segmental, 1 cm osteoperiosteal defects were produced in both radii of 12 rabbits. One side was covered with hydroxyapatite/polylactic acid(HA/PLA) membrane encapsulated as a tube. The contralateral side served as an untreated control. Healing courses were detected by radiographic and histologic examinations. RESULTS: All control sides showed nonunion, whereas there were consistent healing pattern in test sides. CONCLUSION: Membrane technique can promote bone regeneration. PMID- 12080780 TI - [Study of the bone formation and osteogenesis after transplantation of human periosteal mesenchymal stem cells]. AB - OBJECTIVE: To study the bone formation and osteogenesis after transplantation of human periosteal mesenchymal stem cells(PMSC). METHODS: Suspension of PMSC which obtained from cell culture of periosteal segments in vitro were injected into the backs of nude mice subcutaneously, and the fracture site of neck of femur in old person. RESULTS: Subdermal nodules were observed by naked eyes after 11 days of transplantation. 4 weeks later, their anatomic diameter reached 2-7 mm(averaged 3.6 mm). It was proved that the subdermal nodules were trabecular ball trapped with fibrous tissue. The nodules were investigated by human special apoB gene with PCR, and the test of anti-human-tissue precipitin reaction(AHTPR). The results of PCR and AHTPR were positive reaction. There were no subdermal nodules formed in the sites of injection of frozen-melted PMSC or culture medium. The new callus in the sites of fracture were tested by PCR test, and two kinds of apoB gene products were detected. CONCLUSION: The results indicated that the implanted PMSC could form new bone directly in nude mice, and the cells of donor and recipient all could form new bone. PMID- 12080781 TI - [The experimental study of repairing bone defects with allogeneic bone matrix gelatin and plaster]. AB - OBJECTIVE: To study the function of the composite of bone matrix gelatin(BMG) and plaster in the repairing process of bone defects. METHODS: Sixteen New Zealand rabbits which were defected in corpus radii were made as implant zone of bone. Sixteen sides of radii were implanted with the composite of BMG and plaster as experimental group. Others were implanted with BMG(8 sides) and bone stored in alcohol(8 sides) as control groups. The repairing process in bone defects were observed by X-ray and histological examination. RESULTS: There was an obvious osteogenesis in experimental group. The defects of radii were almost healed at 12th week after operation. There were osteogenesis in both control groups, but the repairing process was slower than that of the experimental group. CONCLUSION: The composite of BMG and plaster is a good material for bone transplantation. PMID- 12080782 TI - [Experimental study on osteoinduction of coral composited artificial bone]. AB - OBJECTIVE: To improve the osteoinduction of coral and provide a perfect bone graft substitute for clinical bone defects. METHODS: By combining coral with collagen and recombinant human bone morphogenetic protein-2(rhBMP-2), coral/collagen/rhBMP-2 composite was obtained. The composite was implanted into the back muscle pouches of mice, and coral/collagen or coral/rhBMP-2 were implanted as control. The osteoinduction of the composite was assessed by histology and image analysis system. RESULTS: The chondrocyte differentiation and matrix formation were observed in local sites after one week, lamellar bone with bone marrow were formed after 4 weeks, and coral were absorbed partially. The quantity of osteoinduction was time-related and rhBMP-2 dose-related(P < 0.01). Coral/collagen and coral/rhBMP-2 implants did not show any bone or cartilage formation. CONCLUSION: The coral/collagen/rhBMP-2 composite possesses a superior osteoinduction and will be a new type of bone substitute to be used in orthopedic and maxillofacial surgery. PMID- 12080783 TI - [Applied anatomy of bone flap pedicled with spina scapular branch of transverse cervical artery for mandibular reconstruction]. AB - OBJECTIVE: To study the anatomical basis of vascularized spina scapular bone flap, which was used in mandibular reconstruction. METHODS: Fifteen adult cadavers were adopted in this study. The two common carotid arteries of each cadaver were intubed and perfused with red emulsion respectively. Then the course and distribution of the transverse cervical artery(TCA) and its spina scapular branches were observed on 30 sides. RESULTS: The TCA was divided into two segments: the cervical segment originated from the origin of the artery to the superior margin of the trapezius muscle, and the dorsal segment originated from the superior margin of the trapezius muscle to the site where the TCA bifurcated into the superficial and deep branches. The average length and original caliber of the cervical segment were(4.7 +/- 0.1) cm and (4.0 +/- 0.1) mm. The average length and original caliber of the dorsal segment were (5.88 +/- 0.63) cm and (3.30 +/- 0.35) mm. 86.7% spina scapular branches originated from the superficial branch of TCA and 13.3% from TCA. The length of the spina scapular branch was (4.97 +/- 1.68) cm and its external diameter was (2.08 +/- 0.27) mm. It constantly sent 4-8 periosteal branches to spina with 0.20-1.25 mm in caliber. CONCLUSION: The spina scapular branch of TCA is one of the main blood supplier to the spina scapular area. The spina scapular flap pedicled with spina scapular branch of TCA may provide a new operation for mandibular reconstruction, whose circumpoint locates at the origin of the dorsal segment and the average length of the pedicle is 10.85 cm which enough to transposite to mandibular area. PMID- 12080784 TI - [Experimental study and clinical application on osteogenesis of percutaneous autogenous bone marrow grafting in bone defects]. AB - OBJECTIVE: To observe the osteogenesis of percutaneous autogenous bone marrow grafting in cicatricial bone defect, to seek a good method for treating fracture nonunion. METHODS: Eighteen rabbits were adopted in this study. 1 cm bone defect model was made in each side of radius, 6 weeks later, 2 ml autogenous bone marrow was injected in the right radial bone defect as experimental group, 2 ml autogenous peripheral blood in the left side as control group. X-ray features, histologic changes, Ca and P content in the site of bone defect were studied in various times. Also 15 patients were treated clinically for the nonunion fracture, the average time from nonunion to bone marrow grafting was 13 months. RESULTS: In experimental group, the increasing new bone tissue were observed in X ray and histologic examination. While in control group, no osteogenesis was observed. Ca and P content of experimental group was higher than that of control group. For the 15 patients, 13 cases healed in 5-9 months, 2 cases failed. CONCLUSION: Percutaneous autogenous bone marrow grafting is capable of osteogenesis in the cicatricial bone defects. It can be used in nonunion cases which are not fit for operation of bone grafting because of poor condition of the skin. PMID- 12080785 TI - [Application of repairing tibia and soft tissue defect with free fibula combined tissue grafting]. AB - OBJECTIVE: To investigate a good method for repairing the long bone defect of tibia combined with soft tissue defect. METHODS: From 1988-1998, sixteen patients with long bone defect of tibia were admitted. There were 12 males, 4 females and aged from 16 to 45 years. The length of tibia defect ranged from 7 cm to 12 cm, the area of soft tissue defect ranged from 5 cm x 3 cm to 12 cm x 6 cm. Free fibula grafting was adopted in repairing. During operation, the two ends of fibular artery were anastomosised with the anterior tibial artery of the recipient, and the composited fibular flap were transplanted. RESULTS: All grafted fibula unioned and the flap survived completely. Followed up for 6 to 111 months, 14 patients acquired the normal function while the other 2 patients received arthrodesis of the tibial-talus joint. In all the 16 patients, the unstable ankle joint could not be observed. CONCLUSION: The modified method is characterized by the clear anatomy, the less blood loss and the reduced operation time. Meanwhile, the blood supply of the grafted fibula can be monitored. PMID- 12080786 TI - [Application of fetal bone in repairing the defected part of osteoma resection]. AB - OBJECTIVE: To investigate the clinical results used fetal bone repairing the defect after osteoma resection. METHODS: In 8 hours after abortion, the fetal bones of four limbs were cut and kept in the normal saline with gentamycin. Then the graft was storaged under 6 degrees C-8 degrees C and the longest storage time was 48 hours. Since 1990, seven cases of osteoma had been cured with fetal bone graft. Among them, there were 5 males and 2 females, aged from 6 to 38 years, the bone defect volume ranged from 5.5 cm x 3.6 cm x 2.5 cm to 9.0 cm x 4.3 cm x 3.2 cm. During operation, the osteoma was resected radically, then the defected cavity was impacted with prepared fetal bone. RESULTS: After operation, the general condition was good, all the incision wound healed primarily. Sixteen weeks after operation, the defected cavity was repaired completely and the medullary cavity reopened. Follow-up for 8 months to 5 years, no recurrence or side-effect were observed. CONCLUSION: Because of the more rapid expansion and replacement effect, the stronger osteoinduction, and the lower immune response, the fetal bone is practical material for repairing the bone defect in osteoma resection. PMID- 12080787 TI - [A surgical treatment of early partial closure of the epiphysial plate]. AB - OBJECTIVE: To introduce the technique of epiphysiolysis and epiphysis grafting in the treatment of early partial closure of the epiphysial plate in children, and evaluate its effect. METHODS: Retrospective study was performed in 10 cases of cubitus varus and valgus, or genu varum and valgum due to early partial closure of the epiphysial plate, 6 cases received simple cuneiform osteotomy, and 4 cases received epiphysioloysis and epiphysis grafting. RESULTS: Clinical outcoming after 6-month to 5-year's follow-up showed 1 case deformity of recurrence and 2 cases of non-isometric limb after simple osteotomy, while no recurrence and isometric limb after epiphysiolysis and epiphysis grafting. CONCLUSION: Epiphysiolysis and epiphysis grafting may effectively prevent the recurrence of postoperative deformity, and restore the longitudinal growth of limb. PMID- 12080788 TI - [Application of skin-stretching devices in reparative and reconstructive surgery]. AB - OBJECTIVE: To testify the availability and the clinical efficacy of the skin stretching device. METHODS: The skin-stretching device designed by ZHOU Li-an, which was used in the treatment of the granulation wound, the donor site of flap, and the skin defect after the excision of tumor by skin traction technique at the wound edges. RESULTS: The usage time of skin-stretching device was 15-720 minutes, it could be used preoperatively, intraoperatively and postoperatively. Followed up 16 cases for 6 to 12 months, the device was effective. CONCLUSION: The skin stretching device is practical and effective in wound closure of skin defect. PMID- 12080789 TI - [The use of subdermal vascularized thin flap in the repair of the burned scar on neck]. AB - OBJECTIVE: To evaluate the clinical efficacy of thin flap with subdermal vascular network of the neck-pectoral region on repair of the contracture of the burn scar on the neck. METHODS: From March 1990 to May 1998, 21 cases of deformity of neck due to burn scar were repaired with the thin flap ranging from 8 cm x 5 cm to 14 cm x 8 cm, and all of the cases were followed up for 6 to 42 months. RESULTS: Except partial necrosis of the distal end of the flap in 1 case, the flaps in the other 20 cases all survived and presented a satisfactory appearance and function. CONCLUSION: The thin flap with subdermal vascular network in neck-pectoral region may provide a large area of flap, and could be easily transferred. It's an ideal flap for the repair of skin defect on the neck. PMID- 12080790 TI - [Repair of soft tissue defect on heel with island-shaped fascial flap of lateral foot]. AB - OBJECTIVE: To research the result of defect repairing on heel with island-shaped fascial flap of lateral foot. METHODS: Twelve cases of soft tissue defected on heel, who were resulted from trauma, chronic ulcer, chronic osteomyelitis, squamous carcinoma, and necrosis following frozen injury, were treated by transfer of island-shaped fasical flap of lateral foot. RESULTS: Only 2 patients suffered marginal necrosis of flap in early stage and healed after changing dressing. The others succeeded completely. All the cases were followed up for 8 to 78 months. There was no recurrence of squamous carcinoma, no fistula or necrosis bone formed. The sensation of the flap recovered. The repaired area was similar to the heel in skin texture. CONCLUSION: For the characteristics of heel skin, the transfer of island-shaped fascial flap of lateral foot has the following advantages: Similar structure of skin, reliable nerve and blood supply, simple operative techniques, and large area of donor flap. PMID- 12080791 TI - [Observation of reconstruction of wrist joint by replacement of distal end of radius with vascularized fibular head]. AB - OBJECTIVE: This paper is aimed to observe the long-term result of reconstruction of wrist joint by replacing distal end of radius with vascularized fibular head in patient of giant tumor. METHODS: From July 1978 to November 1993, 12 cases of giant tumor of distal end of radius were treated by this procedure and followed up for 6 months to 15 years, and the movement range of wrist, grip strength, and pain were evaluated. RESULTS: The painful wrist released in 11 cases, only one case still existed mild pain and felt discomfortable. The average range of movement of wrist joint was 34.05 degree in volar flexion, 52.80 degree in dorsal extension, 26.25 degree in ulnar deviation, and 19.75 degree in radial deviation, and average grip strength was 44.6 kg. Compared with the contralateral side, there were accounted for 46.2%, 72.7%, 76.1%, 80.6%, and 76.7% respectively. CONCLUSION: Replacement of distal end of radius with fibular head to reconstruct wrist joint can restore function of carpal joint, which is proved to be a safe and effective method. PMID- 12080792 TI - [A review on the application of myoblast on gene therapy and tissue engineering]. AB - OBJECTIVE: Because of its special biological characteristics, myoblast might play a role in gene delivery and cell-to-biomaterial interactions. In this paper, the biological features of myoblast and its application on gene therapy and tissue engineering was discussed. METHODS: Documents about proliferation and differentiation of myoblast were reviewed in details. The prospects of its application on gene therapy and tissue engineering were also presented. RESULTS: Myoblast was important in muscle regeneration. The activation of myoblast to proliferate and differentiate was the very beginning of regeneration after injury. The cultured myoblast had high potential to proliferate, it was ready to fuse with each other and to form myotube (the special behavior of myoblast differentiation). Myoblast transplantation had been studied as a possible treatment for inherited myopathies, such as Duchenne muscular dystrophy. The transplanted myoblast could fuse with host myofibers, so the delivered target gene integrated into host. Several myoblast-mediated gene delivery system had been established, including the gene delivery of human factor IX (hFIX), erythropoietin (EPO) and clony stimulating factor-1 (CSF-1). Results from animal experiments demonstrated that myoblast-mediated gene delivery could be used as gene therapy for some inherited diseases. And recently, some authors have shown great interest in the interaction between myoblast and type I collagen gels. It was found that myoblast could keep on proliferating and differentiating in collagen gels and could form discoid, tubular materials. CONCLUSION: Myoblast has great importance in gene therapy and tissue engineering. It is suggested that more efforts should be made in this field. PMID- 12080793 TI - [Comparative study of the character of cefazolin pharmacokinetics between Neijiang pig and human]. AB - OBJECTIVE: To study the character of cefazolin (CEZ) pharmacokinetics on Neijiang pig and human. METHODS: The serum concentration of CEZ in 8 normal Chinese adult men, 9 of 8-month male Neijiang pigs and 5 of 4-month male Neijiang pigs were detected by high-performance liquid chromatography (HPLC). RESULTS: The pharmacokinetic parameters suggested that two-compartment model was found in all groups after intramuscular injection of CEZ. In normal men, 8-month pigs and 4 month pigs, the peak time (Tmax) was (58.8 +/- 13.0), (19.7 +/- 9.9) and (18.2 +/ 8.6) min respectively, T1/2 alpha was (42.3 +/- 19.7), (19.0 +/- 7.7) and (9.3 +/- 1.9) min, the peak concentration (Cmax) was (101.6 +/- 14.6), (28.7 +/- 9.0) and (23.5 +/- 4.6) mg/L; Vd was (0.096 +/- 0.016), (0.374 +/- 0.184) and (0.386 +/- 0.211) L/kg; T1/2ka was (22.5 +/- 6.8), (8.6 +/- 4.8) and (10.6 +/- 10.2) min; T1/2 beta was (117.3 +/- 8.6), (84.2 +/- 9.8) and (45.1 +/- 11.5) min; clean rate of plasma Cl was (0.8 +/- 0.1), (6.8 +/- 1.2) and (11.0 +/- 3.0) ml/kg.min; AUC was (21,803 +/- 4,145), (2,407 +/- 443) and (1,636 +/- 685) mg.min/L. CONCLUSION: It could conclude that the Neijiang pigs could eliminate CEZ effectively, but the absorption, distribution and elimination of CEZ in pigs were quicker than that of in human while the absorption from muscle in both pig groups were lower than that in human. PMID- 12080794 TI - [A review on the treatment of spastic cerebral palsy with selective posterior rhizotomy]. AB - OBJECTIVE: To inquire the indications, contraindications, and operative methods of the treatment of spastic cerebral palsy with selective posterior rhizotomy (SPR), and to improve the therapeutic results and decrease the incidence rate of complications. METHODS: The documents about SPR were extensively consulted, and the mechanisms, indications, contraindications, operative methods, muscular tension changes and complications after operation were reviewed. RESULTS: With the methods of SPR, I alpha fibers of afferent nerve were selectively amputated, reflex circle of spinal cord was locked, and the muscular tension was decreased, so myospasm was removed. The results after operation and incidence rate of complications were closely related to the indications. The therapeutic results were better when the percent of spinal nerve rhizotomy was less than 50%. CONCLUSION: It is a good method for the treatment of spastic cerebral palsy. PMID- 12080795 TI - [Observation on the degeneration and regeneration of the Meissner's corpuscles in the monkey's denervated fingers following sensory nerve implantation under electron microscope]. AB - OBJECTIVE: To observe the degeneration and regeneration of the Meissner's corpuscles after implanted sensory nerve into the denervated monkey's fingers under electron microscope. METHODS: The two finger nerves of the monkey's fingers were denervated. Afterwards, one finger nerve was cut off, and the other was reimplanted into the denervated finger. After 1, 3, 5, 8 and 12 months, the finger skin was cut off and observed under electron microscope. RESULTS: The degenerative changes of nerve ending in Meissner's corpuscles were observed after 1 month of denervation, and the basic structure of the corpuscles had no obvious changes. After 3 months, the axons of corpuscles were disappeared, and the volume of corpuscles was shrunk. The basic structure of nerves was disappeared, and the lemmocyte and neurolemma plate were changed after 5 months. The collagen fibrils in the corpuscles were gradually increased in 8 months, the endoneurial structure and interneurial matrix were completely disappeared and replaced by collagen fibrils in 12 months. After 3 months of nerve implantation, unmyelinated nerve fibers were appeared and grew into the corpuscles. A part of corpuscles innervated in 5 months. Most of corpuscles innervated and myelinated nerve fibers were observed in 8 months. And in 12 months, corpuscles innervated to normal level. CONCLUSION: The implantative sensory nerve by means of reinnervating the original corpuscles and regenerating new corpuscles could innervate the degenerative Meissner's corpuscles. PMID- 12080796 TI - [The separation and detection of the bioactive proteins in nerve regeneration conditioned fluids]. AB - OBJECTIVE: To understand the biological activities of the nerve regeneration conditioned fluid (NRCF). METHODS: Nerve regeneration chamber was made by using silicone tube bridging distal and proximal ends of severed SD rat's sciatic nerve. The biological activities of the proteins in NRCF, which were separated by natural polyacrylamide gel electrophoresis (PAGE), were analysed by being cocultured with excised neonatal dorsal root ganglia (DRG). RESULTS: Eight separated protein bands of NRCF were observed between 67-669 ku in molecular weight, and the protein bands between 232-440 ku showed strong neurotrophic and chemotactic function. CONCLUSION: NRCF has the promoting effects on nerve regeneration. PMID- 12080798 TI - [Direct gene transfer into rabbit peripheral nerve in vivo]. AB - OBJECTIVE: To probe the possibility of direct transfer of exogenous gene into peripheral nerve and its following expression in vivo. METHODS: The PCMV beta plasmid containing cytomegalovirus (CMV) promoter and Escherichia Coli (E. Coli), beta-Galactosidease (beta-Gal) structural gene (lacZ gene) was constructed and injected into the rabbit sciatic nerve. The control group was injected PBS solution. The injected nerves were sampled and tested by beta-Gal enzyme activity assay of the 5-bromo-4-chloro-3-indolyl-beta-D-galactoside and beta-Gal histochemical stain. RESULTS: In the control group, no beta-Gal enzyme activity was detected in the different stages after operation, and beta-Gal histochemical stains showed positive. In the experimental group, enzyme activity could be detected from 2 days to 30 days after operation, and the histochemical stains showed negative. CONCLUSION: The exogenous gene can be transferred into peripheral nerve and expressed with bioactivity, thus the gene therapy to accelerate the recovery of nerve is practical. PMID- 12080797 TI - [Experimental study of morphology and electrophysiology on denervated skeletal muscle]. AB - OBJECTIVE: In order to provide the scientific basis to find out a practical and effective method to evaluate the degree of muscle atrophy and a better method of prevention and treatment of skeletal muscle atrophy. METHODS: Forty-two adult Spray-Dawley rats were used and the model of denervated gastrocnemius muscle was established by cutting off the tibial nerve. The muscle wet weight, diameter and cross section area of myocyte were measured. The motor end-plate, fibrillation potential amplitude and frequency of denervated skeletal muscle were observed. RESULTS: The muscle wet weight rapidly reduced within 4 weeks. Afterwards, it maintained about 30 per cent of normal value, and the diameter and cross section area of myocyte progressively reduced. The motor end-plate slightly changed within 4 weeks, but its degeneration accelerated in 6 weeks and disappeared after 16 weeks. The fibrillation potential amplitude was maximum at 2 weeks and it progressively reduced after 12 weeks of muscle denervation. The changes of amplitude and frequency were consistent with the degeneration of end-plate. CONCLUSION: The muscle wet weight, diameter and cross section area of myocyte, fibrillation potential amplitude and frequency could be considered as the morphological and electrophysiological indexes of muscle atrophy degree. It's suggested that the repairing operation of peripheral nerve should be performed before the disappearance of motor end-plate. PMID- 12080799 TI - [Immunohistochemical study of S-100 protein in degenerative nerve after different pathological brachial plexus injuries]. AB - OBJECTIVE: To explore the regularity of the change of S-100 protein in degenerative nerve after different pathological brachial plexus injuries. METHODS: Eighty SD rats were randomly divided into two groups, right C5, C6 preganglionic injury, and postganglionic injury. The distribution and content of S-100 protein in distal degenerative nerve were detected after 1, 2, 3 and 6 months of injury by immunohistochemical methods. RESULTS: The S-100 protein was mainly distributed along the axons. The S-100 protein positive axons of each time interval decreased after operation, with significant difference from normal nerves (P < 0.01). There was no statistically significant difference among 1, 2, 3 and 6 months group (P > 0.05). The S-100 protein stain of postganglionic group was negative. CONCLUSION: In preganglionic injury, the functional expression of Schwann's cells in the distal stump keeps at a certain level and for a certain period. Since Schwann's cell has inductive effect on nerve regeneration, it suggests that the distal nerve stump in preganglionic injury can be used as nerve grafts. PMID- 12080800 TI - [Applied anatomy of the paraumbilical flap with the lateral anterior branch of the thoracic nerve]. AB - OBJECTIVE: To provide the anatomical basis for the free paraumbilical flap with sensory nerve. METHODS: The morphology, branch and distribution of the inferior epigastric artery and inferior intercostal nerve were dissected and measured in 20 adult cadaver specimens. RESULTS: The diameter of inferior epigastric artery at the original point was (2.3 +/- 0.3) mm, and that of its accompanying vein was (3.6 +/- 0.4) mm. The anterial branch of inferior intercostal nerves transversed through their corresponding intercostal spaces of axilla anterior line and ran out of the superficial fascia at the midclavicular line. The lateral anterior branch of the eighth to tenth intercostal nerves ran out of superficial fascia in the range of 0-7 cm above umbilicus and innervated the paraumbilical flap. CONCLUSION: It is possible to design sensory paraumbilical flap with the lateral anterior branch of the eighth to tenth intercostal nerve. PMID- 12080801 TI - [Repair of recurrent laryngeal nerve injuries after thyroidectomy]. AB - OBJECTIVE: In order to investigate the opportunity of repair and prognosis of recurrent laryngeal nerve injuries after thyroidectomy. METHODS: Twelve cases with recurrent laryngeal nerve injuries after thyroidectomy were immediately and delayed operated on nerve repair and reinnervation. In immediate operation, 5 cases were repaired by direct recurrent laryngeal nerve suture, and 1 case was treated by transposition of the phrenic nerve to the recurrent laryngeal nerve and sutured the adductor branch to the branch of ansa cervicalis. In delayed operation, 3 cases were treated by anastomosis the main trunk of ansa cervicalis to the adductor branch of recurrent laryngeal nerve, and 3 cases were operated on neuromuscular pedicle to reinnervate posterior cricoarytenoid muscle. RESULTS: Followed up 6 months, the effect was excellent in 1 case who was immediately operated by selective reinnervation of the abductor and adductor muscles of the larynx, better in 9 cases, and poor in 2 cases who were delayed operated over 12 months. CONCLUSION: It can be concluded that the earlier reinnervation is performed, the better prognosis is. PMID- 12080802 TI - [The emergent repair of peripheral nerve injury of the wrist]. AB - OBJECTIVE: To investigate the effect of the emergent repair of peripheral nerve injury of the wrist. METHODS: From July 1993 to December 1997, 17 cases were admitted, which 21 injured peripheral nerves were repaired emergently. Among them, there were 11 cases of median nerve injury, 2 cases of ulnar nerve injury and 4 cases of median and ulnar nerve injury. All the nerves were ruptured completely except one which was partially ruptured. The emergent operation was taken and the injured nerves were repaired by microsurgical technique. RESULTS: Followed up 6 to 18 months after operation, 95.25% injured nerves had good outcome. CONCLUSION: Because of the specific structure of the wrist, nerve injury at this part need to be repaired emergently. It can enhance the regeneration of the injured nerve, preserve the function of the intrinsic muscle of hand, and decrease the local adhesion. PMID- 12080803 TI - [Compression of the palmar cutaneous branch of the median nerve at the wrist]. AB - OBJECTIVE: To study the compression factor and clinical manifestation of the compression of the palmar cutaneous branch of the median nerve. METHODS: Anatomic study was done on both sides of 2 cadavers and 6 cases of hand injury in the debridement, the origin, course, branch of the palmar cutaneous branch of the median nerve were observed. From 1995 to 1998, 12 patients of compression of the palmar cutaneous branch were treated by local blockade injection. Among them, there were 8 males and 4 females, aged from 23 to 65 years and the course of disease ranged 3 to 12 months. RESULTS: The palmar cutaneous branch of the median nerve was (1.3 +/- 0.1) mm in diameter, it could be pulled when the wrist dorsi extension. All cases showed good recovery of hand function and no recurrence after 4 to 12 months follow-up. CONCLUSION: The palmar cutaneous branch compression syndrome is closely related to the local anatomy. The diagnosis is definite according to the clinical symptoms and signs, and local blocking is effective on the most patients. PMID- 12080804 TI - [Compression of the deep branch of ulnar nerve at the wrist]. AB - OBJECTIVE: To investigate the compression factor and clinical manifestation of the compression of deep branch of the ulnar nerve at the wrist. METHODS: Anatomic study was done on both sides of 10 cadavers, the deep branch of ulnar nerve, the Guyon's canal and the flexor digiti minimi brevis pedis were observed. Then from Jan. 1990 to Jan. 1997, 5 patients with compression of the deep branch of ulnar nerve at the wrist were treated clinically. Among them, there were 4 males and 1 female, aged from 37 to 48 years and the course of disease ranged from 1 to 5 months. RESULTS: The motor branch of the ulnar nerve passed under the tendinous arcade of flexor digiti minimi brevis pedis. Occasionally, the branch of ulnar artery overpassed the motor branch. Clinically, the tendinous arcade compressed the motor branch was released, and after 2 to 4 years follow-up, the clinical results were satisfactory. CONCLUSION: The main compression factor of the ulnar nerve at the wrist is the tendinous arcade of the flexor digiti minimi brevis pedis, the tendinous arcade should be released sufficiently during the operation. PMID- 12080805 TI - [Effect of L-arginine supplementation on partial-thickness burned patients]. AB - OBJECTIVE: L-arginine is a semiessential dibasic amino acid for humans and animals. This paper aims to investigate the therapeutic effect of L-arginine supplementation on partial-thickness burned patients. METHODS: A randomized controlled clinical trial was designed to evaluate the cellular immune function (T cell count, ratio of CD4/CD8, natural killer cell activity and IL-2 level) and protein metabolism (transferrin, prealbium and nitrogen balance) of patients in the experimental group which daily given 15 g arginine and the control group which daily given 25 g glycine. RESULTS: The natural killer cell activity and IL 2 production in the experimental group were higher than that of the control group. The suppression of transferrin and prealbium was alleviated and the nitrogen balance was improved in the experimental group. CONCLUSION: It suggests that exogenous arginine supplementation is beneficial for recovery of cellular immunity function and protein anabolism in partial-thickness burned patients. PMID- 12080806 TI - [Musculocutaneous flap containing upper part of bilateral trapezius muscles in the treatment of cicatricial contracture deformity of neck]. AB - OBJECTIVE: To evaluate the efficacy of musculocutaneous flap containing the upper part of bilateral trapezius muscles in the treatment of cicatricial contracture deformity of neck. METHODS: From January 1990 to January 1997, twelve cases were treated by musculocutaneous flap containing upper part of bilateral trapezius muscles from 14 cm x 16 cm to 28 cm x 11 cm in size, and 9 of the 12 cases were followed up for 2-3 years. RESULTS: Primary healing of the wounds was observed in all of the cases. The function and the external appearance were satisfactory, and no graft contracture was found in the 9 cases followed up. CONCLUSION: The musculocutaneous flap grafting was an ideal method in the repair of soft tissue defect of neck, with the advantage of invariable blood supply and simple operative procedure. PMID- 12080807 TI - [Transposition of expanded superficial cervical artery skin flaps for the repair of cicatricial contracture of neck]. AB - OBJECTIVE: In order to reduce the complication of the transposition of superficial cervical artery skin flap for the repair of neck defect, the method of pre-expanded skin flap was designed, and its clinical result was observed. METHODS: From March 1995 to October 1997, 12 cases with cicatricial contracture of the neck were treated by the following methods, preexpanded superficial cervical artery skin flap, and then transposed it for the reconstruction the cervical scar after burns. There were 8 males and 4 females and the age ranged from 6 to 32 years. A maximal size of flap was 35 cm x 14 cm and a minimal size was 16 cm x 7 cm. RESULTS: All the flaps were survived except one, which partial necrosis occurred in the tip. Postoperative follow-up for 6 months to 3 years showed that the physiological angle of cervico-mandibular angle was recovered and the appearance of flap was satisfactory without swelling and contracture. CONCLUSION: The preexpanded superficial cervical artery skin flap has many advantages, and it is particularly suitable for reconstruction of severe cervical contracture after extensive burns. PMID- 12080808 TI - [The thumb reconstruction by transferring the injured index finger with pedicles]. AB - OBJECTIVE: In order to inquire the methods of thumb reconstruction by transferring the index finger with incomplete conditions of nerve or blood vessels. METHODS: From April 1987 to October 1997, 6 cases were treated by 3 kinds of operative methods according to the damage type of thumb and complications injures of the rest of hand: 1. transferring the index finger with pedicle without proximal phalanx, 2. transferring the index finger with palmar nerve and blood vessels, and dorsal skin pedicle, 3. transferring the index finger with compound pedicle. RESULTS: All 6 cases of thumb reconstruction were successful. Followed up 6 months to 2 years, the pinching and gribing functions in 6 cases were completely recovered, and the sensation were partly recovered. CONCLUSION: The operative method of thumb reconstruction had following advantages: Simple operation, high survival rate and certain function recovery. It can enlarge the indications of thumb reconstruction. PMID- 12080809 TI - [Membranectomy with intestinal-plasty for the treatment of duodenal and upper jejunal constrictive abnormalities]. AB - OBJECTIVE: To sum up the experience of diagnosis and treatment of intrinsic upper gastro-intestinal membrane, 13 cases in children were studied retrospectively. METHODS: There were 10 boys and 3 girls, the major symptoms were vomiting and epigastric distension. Eleven cases were treated by membranectomy with intestinal plasty, and 2 cases were treated by retrocolic side to end duodenojejunostomy. RESULTS: All cases had good results without severe complications. CONCLUSION: The children who have typical symptom of upper digestive tract should be considered duodental and upper jejunal membrane, and should be proved by contrast radiology. The membranectomy with intestinal plasty is the better operative method. PMID- 12080810 TI - [Vulvar reconstruction using gracilis myocutaneous flaps after radical vulvectomy]. AB - OBJECTIVE: To introduce the method using the gracilis myocutaneous flaps to repair of vulvar defect after radical vulvectomy. METHODS: The gracilis myocutaneous flaps were applied in 4 cases with vulvar carcinoma which located vestibule of vulva and bulk, and the carcinoma was large after radical resection to repair vulvar defect simultaneously. RESULTS: The incision of 3 cases was healing by first intention, and 1 case with type II diabetes mellitus was healing by second intention. After followed up 3-6 years, 1 case was recurrent in 2 years after operation, 3 cases were survival without complication. CONCLUSION: Vulvar reconstruction with the use of gracilis myocutaneous flaps in radical vulvectomy could markedly decrease the rate of complication. PMID- 12080811 TI - [The experimental study on optimal cell density and formation time of tissue engineered autologous cartilage]. AB - OBJECTIVE: This paper aims to investigate the suitable cell density and the best formation time of tissue engineered autologous cartilage and to provide theoretical basis and parameters for clinical application. METHODS: The chondrocytes isolated from mini swines' ears were mixed with injectable biocompatible matrix (Pluronic), and the density of cell suspensions were 10, 20, 30, 40, 50, 60, 70 x 10(4)/ml. The chondrocyte-polymer constructs were subcutaneously injected into the abdomen of autologous swine. The specimens were observed grossly and histologically after 6 weeks, and investigated the suitable cell density. Then the chondrocyte-polymer constructs with suitable cell density were transplanted into the abdomen of autologous swine and evaluated grossly and histologically in 1, 3, 6, 9, 15 weeks after transplantation to investigate the best formation time of tissue engineered cartilage. RESULTS: The experiments demonstrated that the tissue engineered autologous cartilage was similar to the natural cartilage on animals with normal immune system in histological characteristics. The optimal chondrocyte density is 50 x 10(6)/ml, and the proper harvest time is the sixth week. CONCLUSION: With tissue engineering skills, we have identified the optimal chondrocyte density and the proper harvest time. PMID- 12080812 TI - [Graft-versus-host disease and xenotransplantation]. AB - OBJECTIVE: The pathogenesis, mechanism, manifestation and diagnosis of graft versus-host disease(GVHD) are reviewed in this article. METHODS: The relevant articles in recent public magazines were reviewed and summarized. RESULTS: It was indicated that GVHD occurred in the conditioned recipients in animal experiments and clinical transplantations. Humoral and cellular factors were involved in GVHD, which could be diagnosed and classified according to their characteristics. CONCLUSION: As a kind of interactions between the host and donor, GVHD are severely harmful to the host. It may also occur in xenotransplantation, where GVHD can be utilized in the studies on transplant immunology, oncology etc. Xenogeneic GVHD is receiving more and more attentions. PMID- 12080813 TI - [Wound healing and growth factors]. PMID- 12080814 TI - [Transforming growth factor beta 1 and interleukin 6 mRNA expression in wound tissues of patients with diabetic ulcers]. AB - OBJECTIVE: To study the relationship between the changes of mRNA expression in wound tissues of diabetic ulcers and tissue repair. METHODS: The mRNA expression of TGF-beta 1 and IL-6 in eight bioptic samples of diabetic ulcers were detected by RT-PCR and pathologic methods, and the surrounding normal skins from the same patients were measured as control group. RESULTS: The mRNA expression levels of TGF-beta 1 were markedly decreased in the diabetic ulcers compared with control group, while the mRNA expression levels of IL-6 were increased at the same reaction conditions. CONCLUSION: The different changes of mRNA expression level of TGF-beta 1 and IL-6 in wound tissue result in low production and decreased activity of TGF-beta 1 and IL-6, which lower the reparative ability of wound tissue. PMID- 12080815 TI - [Continuous delivery of epidermal growth factor to wounds in vivo by genetically modified fibroblasts transfected with a novel chimeric construct]. AB - OBJECTIVE: This paper aims to explore the new method of continuous delivery of epidermal growth factor to wounds by transfected fibroblasts to promote wound repair. METHODS: It was constructed a novel chimeric expression plasmid in which the biologically active portion of the human epidermal growth factor (EGF) gene was fused in-frame to the human granulocyte colony-stimulating factor signal sequence. RESULTS: Clonally selected human fibroblasts transfected with this construct could secrete biologically active EGF. After the transplantation of irradiated gene-transfected fibroblasts suspended in fibrin glue to murine full thickness wounds, EGF could be demonstrated for at least seven days in the wounds, slowly decreasing from initially 470 ng/L to 140 ng/L in 7 days. No EGF was found in the wound at 14 days. CONCLUSION: A single application of irradiated EGF gene transfected fibroblasts to wounds can continuously deliver the transgene in vivo and can be used to administer drugs to the wound bed during the crucial first seven days of wound-healing. PMID- 12080816 TI - [Effects of basic fibroblast growth factor on the healing of cutaneous chronic wounds]. AB - OBJECTIVE: To observe the effects of basic fibroblast growth factor(bFGF) on the healing of cutaneous chronic wounds. METHODS: Twenty-eight cases with thirty three wounds from trauma, diabetes, pressure and radiation injuries were locally treated with bFGF in a dosage of 150 U/cm2 wounds. The healing time of wounds was used to evaluate the treatment results. RESULTS: The healing time in all of chronic wounds were accelerated. All wounds from trauma, diabetes and pressure were healed within 4 weeks and another 2 wounds from radiation injuries were healed over 4 weeks. The healing rate within 4 weeks was 93.9%. CONCLUSION: The results indicate that bFGF can be used as a promoter to accelerate the healing of chronic wounds in clinic. PMID- 12080817 TI - [Clinical study of various growth factors on the improvement of impaired healing ulcers in patients with diabetic disease]. AB - OBJECTIVE: To investigate clinical effects and possible mechanisms of various growth factors on impaired healing ulcers of patients with diabetic disease. METHODS: Seventy-eight patients were divided into three groups; saline control, epidermal growth factor(EGF) experimental group, and platelet-derived wound healing factor (PDWHF) experimental group. General healing conditions, wound closing index, healing rates and histological changes of the patient's ulcer wound were observed during 1-8 weeks after treatment. RESULTS: The wound closing index and healing rate of ulcers were significantly increased in the EGF and PDWHF experimental groups compared with the control group, while the angiogenesis, fibroblast hyperplasia, and collagen deposit were more obvious in EGF and PDWHF experimental groups than that of control group. The promoting effects on wound healing in PDWHF experimental group were better than in EGF group. CONCLUSION: It suggests that local application of certain growth factor alone or various growth factors together is an effective method to improve the condition of impaired healing of diabetic ulcers. PMID- 12080818 TI - [Comparative study of epidermal growth factor and basic fibroblast growth factor on wound healing]. AB - OBJECTIVE: The biological effects of recombinant human epidermal growth factor (rhEGF) and recombinant human fibroblast growth factor (rhFGF) were evaluated on the model of incised wounds in mini pigs. METHODS: Total of 160 incised wounds in 16 mini pigs were divided into two groups (rhEGF group and rhFGF group), each containing 80 wounds. In rhEGF group, 60 incised wounds were treated with different dosages of rhEGF (50, 10 and 0.5 micrograms/wound), and another 20 wounds were treated with solvent as control group. In rhFGF group, all wounds were treated in the same way as described in rhEGF group, the dosages of rhFGF were 150, 90 and 30 U/cm2 respectively. The measurements of cavity volume and area in wound, histological examination were used to evaluate the results of wound healing. RESULTS: The results showed that wound healing was accelerated in all wounds treated with rhEGF and rhFGF. In rhEGF group, the velocity of re epithelialization was faster than that of rhFGF group, however, new granulation tissue in rhFGF was more than that of rhEGF group. CONCLUSION: The results indicate that rhEGF and rhFGF can stimulate wound healing, however, the mechanisms and the biological effects involved in these processes are quite different. It suggests that it is better to use rhFGF in those wounds which need more granulation tissue formation and use rhEGF in the wounds which mainly need re-epithelialization. PMID- 12080819 TI - [Review on transforming growth factor beta and repair of tissue injury]. AB - OBJECTIVE: To review the progress of the relation between transforming growth factor beta (TGF-beta) and repair of tissue injury. METHODS: The recent articles about TGF-beta and tissue reconstruction were extensively reviewed. The gene identification, production, activation of TGF-beta and its role in the repair course of tissue injury were investigated. RESULTS: TGF-beta belongs to a family of multifunctional polypeptides, its gene structure is highly conservative. Many animal models about TGF-beta and tissue injury have been established. The research mainly focuses on the classification, distribution of TGF-beta receptors and their signal pathway. CONCLUSION: TGF-beta plays an important roles in the regulation of repair of tissue injury. PMID- 12080820 TI - [Effect of basic fibroblast growth factor on repairing transected sciatic nerve in rats]. AB - OBJECTIVE: To investigate the effects of basic fibroblast growth factor(bFGF) on repairing transected sciatic nerves in rats. METHODS: The animal models of the transected sciatic nerve of 40 SD rats were established, which divided into 4 groups: normal saline (NS) group, nerve growth factor (NGF) group, bFGF group and normal control group. The epineurium of the transected sciatic nerve was sutured under microscope, then bFGF or NGF was dropped into local sites and injected intramuscularly once a day for 30 days after operation. Functional repair for the transected sciatic nerves was studied by nerve conductive velocity (NCV) and sciatic nerve function index (SFI). RESULTS: As a criterion, the level of the normal control group was regarded as zero, SFI of NS group, NGF group and bFGF group were -114.30 +/- 10.34, -70.50 +/- 11.01, -50.45 +/- 7.82 respectively at 1 month after operation, and they were -54.96 +/- 16.46, -35.21 +/- 10.80, -27.53 +/- 11.23 respectively in 3 months after operation. NCV of bFGF group was significantly faster than NS group and NGF group. CONCLUSION: bFGF can significantly promote the functional repair of injured peripheral nerve, and its effects are better than NGF. PMID- 12080821 TI - [The early protective effects of basic fibroblast growth factor on acute spinal cord injury in rats]. AB - OBJECTIVE: To study the early protective effects of basic fibroblast growth factor(bFGF) on the experimental acute spinal cord injury. METHODS: Thirty-four SD rats were randomly divided into three groups, and were subjected to contusion of thoracolumbar spinal cord. A thin plastic tube was placed in subarachnoid space below the injury level for perfusion. The bFGF-treated rats were received 20 microliters bFGF(containing bFGF 100 U) at once, 30 min, 1, 2, 3, 4, 6, 12, 24 and 48 hours after injury, and an equal volume of normal saline was given to the control group at the same time. The injured spinal cord was detected by morphological observation and biochemical index after injury. RESULTS: The degree of ionic disorder in bFGF-treated rats was significantly ameliorated and the contents of H2O were also markedly decreased. The morphological finding showed that the damages of gray and white matter in bFGF-treated rats were slighter than those of saline-treated rats. CONCLUSION: bFGF has some protective effects on the secondary lesion of early spinal cord injury in rats. PMID- 12080822 TI - [Rescue of motoneuron from brachial plexus nerve root avulsion induced cell death by Schwann cell derived neurotrophic factor]. AB - OBJECTIVE: To study the protective effects of Schwann cell derived neurotrophic factor (SDNF) on motoneurons of spinal anterior horn from spinal root avulsion induced cell death. METHODS: Twenty SD rats were made the animal model of C6.7 spinal root avulsion induced motoneuron degeneration, and SDNF was applied at the lesion site of spinal cord once a week. After three weeks, the C6.7 spinal region was dissected out for motoneuron count, morphological analysis and nitric oxide synthase (NOS) enzyme histochemistry. RESULTS: 68.6% motoneurons of spinal anterior horn death were occurred after 3 weeks following surgery, the size of survivors was significantly atrophy and NOS positive neurons increased. However, in animals which received SDNF treatment, the death of motoneurons was significantly decreased, the atrophy of surviving motoneurons was prevented, and expression of NOS was inhibited. CONCLUSION: SDNF can prevent the death of motoneurons following spinal root avulsion. Nitric oxide may play a role in these injury induced motoneuron death. PMID- 12080823 TI - [Expression and significance of basic fibroblast growth factor in ovarian epithelial neoplasm]. AB - OBJECTIVE: To determine the effect of basic fibroblast growth factor (bFGF) on the biological behaviour of ovarian epithelial neoplasm. METHODS: Ten cases of normal ovarian tissues and eighty cases of ovarian epithelial tumor tissues were detected by immunohistochemical methods. Mias-2000 Picture Analysis System was used to study the relationship of bFGF expression intensity and microvessel count, FIGO stage, pathological grade and classification of ovarian epithelial neoplasm. RESULTS: 1. Expression of bFGF was mainly in cytoplasm and nucleus in several cells of borderline and malignant tumor. 2. The expression intensity of bFGF was closely related to the malignant degree of ovarian epithelial neoplasm. The density of bFGF expression was (3.35 +/- 3.52)% in normal ovarian epithelium, (19.25 +/- 21.73)% in benign tumor, (33.78 +/- 10.86)% in borderline tumor and (48.18 +/- 12.93)% in malignant tumor. The results indicated that bFGF might play an important role in carcinogenesis of ovarian epithelial neoplasm. 3. The expression intensity of bFGF was increased with the FIGO stage of ovarian tumor. 4. The expression intensity of bFGF was increased accompanying with the decrease of differentiation degree in ovian neoplasm. 5. In borderline tumor, expression intensity of bFGF in serous cystadenoma was significantly higher than in mucinous cystadenoma, which indicated bFGF might be an important factor in canceration of ovarian epithelial neoplasm. CONCLUSION: bFGF may play important roles in carcinogenesis, development, invasion and metastasis of ovarian epithelial neoplasm. PMID- 12080825 TI - [Experimental study on stimulation of guided bone regeneration by acid fibroblast growth factor]. AB - OBJECTIVE: To investigate the effect of acid fibroblast growth factor (aFGF) on guided bone regeneration (GBR), to study whether aFGF can promote the repairing ability of GBR in bone defect. METHODS: 10 mm long segmental defects were created in the diaphyses of both radii in 16 New Zealand rabbits. The defect was bridged with a silicon tube. Human recombinant aFGF was instilled into the tube on the experimental side, while the contralateral tube was instilled with saline as control group. The radiographic, gross and histologic examination of the samples were analyzed at 2, 4, 6 and 8 weeks after operation. RESULTS: On the experimental side, there was new bone formation in the bone medullary cavity, the endosteum and the section surface of the cortex at 2 weeks. At 4 weeks, at the center of the blood clot in the tube there was new bone formation and bone defect was completely healed at 8 weeks. On the control side, new bone formation was less in every period compared with that of the experimental side. At 8 weeks, there was only partial healing of the bone defect. CONCLUSION: It can be concluded that aFGF can promote new bone formation and facilitate GBR in bone defect. PMID- 12080824 TI - [Effects of basic fibroblast growth factor on repairing injury of intestinal mucosa in acute necrotic pancreatitis]. AB - OBJECTIVE: To observe the effects of basic fibroblast growth factor (bFGF) on repairing injury of intestinal mucosa in acute necrotic pancreatitis (ANP). METHODS: Sixteen dogs of ANP animal model were made by injection of 5% sodium taurocholate (0.5 ml/kg) with 3,000 U/kg trypsin into the pancreatic duct. The mucosa structure, content of protein, DNA and malondiethylaldehyde (MDA) were observed after ANP and treatment with bFGF, and the plasma lipopolysaccharide and endothelin-1 were detected. The organs of dogs were made to bacterial culture. Ileal mucosa was collected for histological and ultrastructural studies. RESULTS: The results showed that after treatment with bFGF, the injury of intestinal mucosa in ANP was abated. The length, height and area of mucosa microvillus, the content of DNA and protein of ileal mucosa were significantly increased, while the plasma endothelin-1 and lipopolysaccharide were reduced. The organ bacterial translocation rate was also decreased in 50%. CONCLUSION: bFGF has good effects on abating injury of intestinal mucosa, protecting gut barrier function, reducing the incidence of lipopolysaccharide and bacterial translocation after ANP. PMID- 12080826 TI - [Comparative study on osteogenesis of three types growth factors recombination artificial bones]. AB - OBJECTIVE: To compare the osteogenesis of recombination artificial bones, which are bovine deproteined bone (bDPB) and bovine bone morphogenetic protein (bBMP), combined with tumor necrosis factor alpha (TNF alpha), basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) respectively. METHODS: One hundred trephined skull bone defects in fifty rabbits were divided into four groups, which implanted with bDPB/bBMP/TNF alpha, bDPB/bBMP/bFGF, bDPB/bBMP/EGF, and bDPB/bBMP respectively. X-ray and histological changes were observed in the 1st, 2nd, 4th, 6th, 8th weeks after implantation. The content of 35S and 45Ca and ash weight were measured at 10 and 42 days after operation. RESULTS: The osteogenesis of bDPB/bBMP/TNF alpha group was stronger than that of bDPB/bBMP/bFGF group(P < 0.01), while bDPB/bBMP/bFGF group was stronger than that of bDPB/bBMP/EGF(P < 0.01). No significant statistical difference were found between bDPB/bBMP/EGF and bDPB/bBMP(P > 0.05). CONCLUSION: TNF alpha combined with bBMP and carrier can stimulate bone formation and increase the volume of new bone in vivo. It suggests that bDPB/bBMP/TNF alpha is a valuable biomaterial of bone graft. PMID- 12080827 TI - [Guided bone regeneration by using biodegradable co-polymer membranes in rabbits]. AB - OBJECTIVE: To repair long bone segmental defects using biodegradable poly epsilon caprolactone (PCL) and polylactic acid(PLA) co-polymer membranes, and explore its role and mechanism in guided bone regeneration (GBR). METHODS: Rabbit radial segmental defects (1.2 cm in length, retain the periosteum) were created in this study, 24 animals were divided into 2 groups. The membranes were used to enclose the defects in experimental group, and no treatment in control group. After 3, 6, and 12 weeks of operation, X-ray, gross and histological examinations were observed. RESULTS: The bone regeneration of experimental group was better than that of control group. Three weeks after operation, obvious external callus along the membrane were found in experimental group, and bony linking composed of external callus bridge were found in 6 weeks after operation. After 12 weeks of operation, callus bridge outside the membrane and bony reunion inside the membrane were achieved in experimental group. While in control group, typical nonunion was observed after 6 weeks of operation. CONCLUSION: Guided bone regeneration can be achieved by using biodegradable membrane. The defects are repaired by the means of outside membrane callus and relatively late inside membrane callus. The membrane can prevent the ingrowth of fibrous tissue into defect area, thus nonunion are avoid, and keep a high concentration of nutritive elements, also serve as a frame for osteocyte growth to enhance bone healing. PMID- 12080828 TI - [Wound repair with single or double reverse tongue-shaped skin flap]. AB - OBJECTIVE: To seek a simple, safe and effective method for the instant repair of soft-tissue defects after excision of superficial tumor, chronic ulcer and scar constructure as well as injury. METHODS: From August 1993 to October 1997, according to the location and size of the lesion, adjacent single or double reverse tongue-shaped skin flaps were designed to repair the defects of head, face, limb and shoulder in 31 cases, and the donor sites were closed directly. RESULTS: The wounds were healed by first intention and the skin flaps were all survived postoperatively. CONCLUSION: It suggests that the tongue-shaped flap have the following characteristics: abundant blood supply, high survival rate, wide application, less trauma, and adjacent tissue can be fully used. It is a simple, safe and effective method for the instant repair of soft-tissue defects. PMID- 12080829 TI - [Repair of multiple fingers degloving injury with abdominal "s"-type skin flap]. AB - OBJECTIVE: To explore a new surgical management of multiple fingers degloving injury. METHODS: In 1994 to 1997, 47 cases with multiple fingers degloving injury were sutured by two reverse "s"-type skin flaps on abdominal flank. RESULTS: The skin flaps in 46 cases survived and the wounds obtained primary heal. CONCLUSION: The application of abdominal flank "s"-type skin flap is reliable and convenient in the treatment of multiple fingers degloving injury. PMID- 12080830 TI - [Abdominal bipedicled subepidermal vascular-network flap in repair of degloving injury of hand]. AB - OBJECTIVE: To evaluate the clinical result of repairing degloving injury of hand with abdominal bipedicled subdermal vascular-network flap in emergence. METHODS: From 1994 to 1997, 19 cases with degloving injury of hand were treated with two flaps, one flaps with the inferior epigastric artery as pedicle, another with superficial epigastric artery. The two skin flaps were designed oppositely to cover the injured hands. RESULTS: All the flaps survived. Followed up for 1 to 3 years, the contour of hands were fine. Extension of fingers was normal and opponers of thumbs was good. Range of flexion of metacarpo-phalangeal joints was from 45 to 60 degrees, and the proximal interphalangeal joints was from 10 to 25 degrees. CONCLUSION: For the subdermal vascular network, the bipedicled flaps of abdomen in repair of degloving injury of hand have sufficient blood supply, strong resistance to infection, high survival rate, and good contour. PMID- 12080831 TI - [Local plantar rotatory flap for repairing of soft tissue defect of heel]. AB - OBJECTIVE: Soft tissue defect of heel is not uncommon. Transplantation of free cutaneous flap and transfer of axial cutaneous flap have been used in treating such defect successfully, but both of them are somewhat complicated. Local plantar rotatory flap might show great importance in this field. METHODS: Since March 1993 to March 1998, 9 cases with soft tissue defect of heel were repaired by local plantar rotation flaps. The size of defect ranged from 2 cm x 4 cm to 6 cm x 8 cm, and it was designed superficial to plantar fascia. The flap was medially based, and nutrilized by proximal plantar subcutaneous plexus of blood supply as well as lateral and medial plantar nerve. RESULTS: Followed up 4 months to 2 years, all the flaps were survived. Sensation of the flap was preserved in 7 patients, who had normal sensation of the donated area preoperatively. The transferred flap was endurable to body bearing. CONCLUSIONS: The flap is easily prepared with reliable blood supply and sensation of the flap preserved. The method is worthy to be recommended for widely use because of its advantages over other methods. PMID- 12080832 TI - [Reconstruction of hip joint function: old fracture-dislocation of hip joint complicated with deformed healing of upper 1/3 of fractured femur--a case report]. AB - OBJECTIVE: To explore a method of reconstruction of hip joint function after deformed healing of the upper 1/3 of fractured femur as a complication of old fracture dislocation of hip joint. METHODS: A patient with loss of function in hip joint and fusion of knee joint was treated with lock for femur intra medullary fixation in April 1997. RESULTS: Before operation, the diseased hip joint lost its most functions and the entire lower extremity was disabled because the knee joint had been fused. One year after operation, the follow-up examination revealed that the patient could walk by crutches without discomfort, his daily life and work recovered to normal. CONCLUSION: It is effective to treat a patient suffering old fracture-dislocation of hip joint complicated with deformed healing of the upper 1/3 of fractured femur by means of individualized artificial joint replacement and a prosthesis body with lock for femur intra medullary fixation, and it is helpful for the development of a new clinical idea to reconstruct functions in the management of some particular cases. PMID- 12080833 TI - [Clinical observation of phenytoin in promoting fracture healing]. AB - OBJECTIVE: To observe the promoting effect of phenytoin on fracture healing. METHODS: Fourty cases with close fractures of tibia and fibula were included and divided into two groups randomly: the experimental group was administrated with phenytoin and Chinese traditional drug-Jiegudan orally, while the control group was given Jiegudan only. Longitudinal percussion pain, clinical healing time of fracture, growth of calus in X-ray film were detected to evaluate the clinical result. RESULTS: All the fractures were healed in 3 months. But the experimental group was superior to the control group in all indexes. CONCLUSION: Administration of phenytoin orally can markedly promote fracture healing. PMID- 12080834 TI - [Resection of odontoid process by transoccipitocervical posterolateral approach to treat old atlantoaxial dislocation]. AB - OBJECTIVE: To explore a new surgical treatment of old atlantoaxial dislocation. METHODS: 5 patients aged 12 to 45 years, who suffered from old atlantoaxial dislocation for 1 to 4.5 years, received the new surgical treatment by the transoccipitocervical posterolateral approach, and expansion of the foramen magnum, odontectomy and bony fusion of the occipitoatlantoid region were performed. Neurological examinations and CT scan were undertaken after operation. RESULTS: All 5 patients survived. Sensory function recovered almost completely and motor function was better more than 1 degree. CT scan showed the diameter of the spinal canal restored well. CONCLUSION: The transoccipitocervical posterolateral approach is an ideal and safe pathway to reach the site of old atlantoaxial dislocation and the operating field is exposed completely. Anterior posterior decompression and occipitocervical spinal fusion can be performed simultaneously. PMID- 12080836 TI - [Reconstruction of elbow flexion and shoulder abduction with transfer of pectoralis major]. AB - OBJECTIVE: To evaluate the clinical application of primary transfer of pectoralis major to reconstruct the elbow flexion and shoulder abduction. METHODS: 12 cases of old injury of branchial plexus with dysfunction of both elbow and shoulder joints were received surgical operation to reconstruct the palsy joints by primary transfer of pectoralis major, shoulder abduction was reconstructed by clavicular head and elbow flexion by sternal head respectively. All cases were followed up for 5 to 18 months. RESULTS: The function of both joints recovered obviously, the total superior rate is 91.7%. CONCLUSION: Only if the palsy joints, shoulder or elbow, remained normal or almost normal passive motion, and the muscle power of pectoralis major over 4 degrees, the primary transfer of pectoralis major should be a simple, reliable and convenient technique to reconstruct the palsy joints. PMID- 12080835 TI - [Resection of spinal tumor and reconstruction of spinal stability]. AB - OBJECTIVE: Both primary and metastatic tumor of spine can influence spinal stability, spinal cord and nerves. The principles of dealing spinal tumor are resection of tumor decompression on spinal cord and reconstruction of spinal stability. METHODS: Since Aug. 1993 to Oct. 1996, 15 cases with spinal tumor were treated, including 4 primary spinal tumor and 11 metastatic tumor. Tumor foci were mainly in thoracic and lumbar spine. Graded by Frankel classification of spinal injuries, there were 1 case of grade A, 1 of grade B, 3 of grade C, 5 of grade D and 5 of grade E. Tumors of upper lumbar spine and thoracic spine were resected through anterior approach. Posterior approach also was adopted once posterior column was affected. Tumors of lower lumbar spine were resected by two staged operation: firstly, operation through posterior approach to reconstruct spinal stability: secondly, operation through anterior approach. After resection of tumor, the spines were fixed by Kaneda instrument, Steffee plate or Kirschner pins. To fuse the spine, bone grafting was used in benign tumor and bone cement used in malignant tumor. RESULTS: Except one patient died from arrest of bone marrow, the others were followed up for 3 to 20 months. Postoperatively, 11 patients could sit up on one foot with the help of body supporter, and 9 patients could walk in two weeks under careful monitoring. There was no exacerbation of symptom and failure of fixation. The function of spinal cord was improved: 1 case from grade B to grade E, 1 from A to C, 2 from C to E and 4 from D to E. CONCLUSION: The spine can be reconstructed for weight bearing early by internal fixation. The symptom can be relieved and the nervous function can be improved by resection of tumor and decompression. PMID- 12080837 TI - [Application of sartorius muscle in the quadricepsplasty]. AB - OBJECTIVE: Extension stiffness of knee joint is always treated by the quadricepsplasty, but the main deficiency of this method is that patient feels weakness of lower limb and easily kneels down. The aim of this article is to explore the method to resolve the complications after quadricepsplasty. METHODS: Since 1978 to 1997, on the basis of traditional procedures of quadricepsplasty, sartorius muscle was used to reinforce the extension of knee joint. The lower 2/3 of sartorius muscle was fully dissociated only with its insertion intact. A tendon-periosteal-bone flap, about 2 cm in width, was managed on the anterior surface of patella, with its pedicle on the medial edge of patella. The tendon periosteal-bone flap was used to fix the dissociated sartorious into patella to reinforce the extension of knee joint. The very lower part of sartorius was mainly aponeurosis, with the help of an aponeurosis bundle of iliotibial tract, it was fixed into the insertion of patellar ligament, through a bony tunnel chiseled adjacent to the insertion of patellar ligament. By now the movement of knee joint extension was strengthened by the transferred sartorius muscle. Postoperatively, every patient was required to extend and flex knee joint actively and/or passively. Altogether 12 patients were treated, 9 of them were followed up with an average of 14 months. RESULTS: The average movement was increased from 15 degrees to 102 degrees, and the average myodynamia was improved from grade II to grade IV. CONCLUSION: Traditional quadricepsplasty co-operated with transfer of sartorius muscle can strengthen the myodynamia of knee joint extension. It is simple method and can really achieve good function. PMID- 12080838 TI - [Comparison of operation mode and clinical effect of different surgical treatments of gluteus muscle contracture (with 98 cases reports)]. AB - OBJECTIVE: To explore a better surgical mode to treat gluteal muscle contracture. METHODS: In 98 cases, superior cross incision of large tuberosity was performed in 20 cases, posterosuperior curve incision of large tuberosity was performed in 20 cases, and exteroinferior cross incision of iliac spine in the other 58 cases. The patients were observed after operation and followed up. RESULTS: The patients who received exteroinferior cross incision of iliac spine recovered soon without bleeding and infection after operation, which indicated the lowest incidental rate of postoperative complication. CONCLUSION: Exteroinferior cross incision of iliac spine is a safe and effective surgical operation to loosen the gluteal muscle contracture. PMID- 12080839 TI - [Measurement of strength of thigh muscles through arthroscope after reconstruction of anterior cruciate ligament]. AB - OBJECTIVE: To observe the strength of thigh muscles after reconstruction of anterior cruciate ligament by autogenous bone-patellar tendon-bone graft. METHODS: Twenty-three patients, 9 males and 14 females, were followed up one year after reconstruction of the anterior cruciate ligament with autogenous bone patellar tendon-bone graft. Through arthroscope, no intra-articular derangement was found. The strengths of isometric and isotonic contractions of the quadri ceps and the hamstrings muscles of the affected and contralateral thighs were recorded. RESULTS: The donor side for autogenous bone-patellar tendon-bone graft showed significant decrease (P < 0.01), but no effect on that of the hamstrings muscle(P > 0.05). CONCLUSION: To reconstruct the anterior cruciate ligament, harvest of the bone-patellar tendon-bone graft as a reparative material may markedly lower the strength of the quadriceps femoris muscle. PMID- 12080840 TI - [Review on transformed cell and tumorigenicity]. AB - OBJECTIVE: To explore the relationship between characteristics of transformed cell and tumorigenicity. METHODS: Documents about transformed cell and tumorigenicity were reviewed in detail. RESULTS: Normal biological characteristics and cell function could be maintained in non-tumorigenic transformed cell, but it was changed markedly in malignant transformed cell. CONCLUSION: Non-tumorigenic transformed cell can be served as a standard cell line to study the function and growth characteristics of normal cell. PMID- 12080841 TI - [Direct and indirect recognition in pig-to-man xenotransplantation]. AB - OBJECTIVE: To investigate the role of direct and indirect recognition in pig-to man xenotransplantation. METHODS: Taken the peripheral blood lymphocytes (PBLC) from three Neijiang pigs and two humans as stimulators and respondors, the one way mixed lymphatic reactions (MLR) of xenograft were carried out, and allo- and self-PBLC as control. RESULTS: Among the three patterns of MLR, syngeneic was MLR the lowest in proliferation, the allogenic MLR was the highest, and the xenogenic MLR was medium. The PBLCs from humans and pigs were matched on HLA-A, B, DR and DQ by means of modified Terasaki assay. The match on pigs was failure because of the pre-existing natural xenogenic antibody in the testing serum. CONCLUSION: The results suggest that the degree of MHC matching still affect the rejection in xenotransplantation, but the present serum assay of MHC matching is not fit for pig. PMID- 12080842 TI - [Establishment of the animal models of mandibular distraction osteogenesis]. AB - OBJECTIVE: To establish the animal models of mandibular distraction osteogenesis in rabbits and study its osteogenetic mechanism. METHODS: The right mandibles just anterior to the first molars of 12 rabbits were performed osteotomies, and the mandibles were positioned with distractors. The left mandibles were control group without operation. After 1 week, the distractors were stretched 0.9 mm every day for 10 days progressively. One day, 2, 4, 8 weeks after distraction, the mandibles were studied with gross measurement, X-ray, and histological examination. RESULTS: The right mandible were lengthened 8.3 mm on average without bone nonunion and deformity healing. It was observed that the gaps between the distracted bone edges were first occupied by fibrous tissue. Two weeks after distraction, it was found that the gaps were bridged by callus in X ray, the new bone and the normal bone could not be differentiated clearly after 8 weeks. In histological sections, there were collagen bundles in early distraction, then those collagen bundles were calcificated and become trabeculaes. No Cartilage was found during distraction. CONCLUSION: It suggests that the rabbit mandible can be lengthened by distraction osteogenesis, and the new bone is formed by intramembranous ossification. PMID- 12080843 TI - [Effect of chitosan membrane on tendon adhesion and healing]. AB - OBJECTIVE: To investigate the effects of chitosan membrane on tendon adhesion and healing and obtain experimental data for clinical use in preventing postoperative tendon adhesion. METHODS: The long flexor tendon of 55 adult legborn hens were cut and sutured encapsulated by chitosan membrane. Movement and anti-tension capability of tendon were assessed at 2, 4, 6, 8 and 10 weeks after operation by SWD-10 type tendon stretcher. Tendon healing and adhesion were observed with light microscope. RESULTS: Tendon healing could be effected by chitosan membrane within 4 weeks, and tendon anti-tension strength was increased after 4 weeks. CONCLUSION: Chitiosan membrane possesses the following characteristics: no side effects, good permeability, resolvable, absorbable and selective inhibition the growth of fibroblast. It is a desirable biological material to prevent tendon adhesion. PMID- 12080844 TI - [The effect of combined treatment of recombinant human growth hormone and insulin like growth factor-1 on wound healing and protein catabolism in burned rats]. AB - OBJECTIVE: To investigate the effect of combined treatment of recombinant human growth hormone (rhGH) and insulin-like growth factor-1 (IGF-1) on wound healing and protein catabolism in burned rats. METHODS: Forty Wistar rats with deep II degree scald injury were divided randomly into four groups and received rhGH (0.1 U/kg.d), rhGH (0.1 U/kg.d) plus IGF-1 (2.0 mg/kg.d), IGF-1 (2.0 mg/kg.d) and Ringer's solution (2 ml/kg.d, as control group) respectively. The wound healing time and protein catabolism levels of every groups were compared after 2 weeks. RESULTS: Total body weight began to increase after 2 weeks in rhGH group and rhGH plus IGF-1 group, but in control group, it was occurred after 4-5 weeks. The body weight of rhGH plus IGF-1 group was 1.65 times than that of rhGH group. The wound healing time in rhGH plus IGF-1 group (17.1 +/- 4.4) days was significantly lower than that of rhGH (20.5 +/- 4.8) days and control group (29.7 +/- 6.3) days. The protein level of rhGH plus IGF-1 group was significantly higher than that of control group and rhGH group. CONCLUSION: It suggests that rhGH plus IGF-1 with synergism is more effective in promoting wound healing and increasing the protein catabolism. PMID- 12080845 TI - [Prevention of postoperative deep venous thrombosis in lower limb after operation by intermittent pneumatic compression]. AB - OBJECTIVE: To investigate the effect of intermittent pneumatic compression on prevention of deep venous thrombosis after operation of lower limbs. METHODS: From Oct. 1997 to Aug. 1998, forty cases were received Doppler examination preoperatively, which showed no deep venous thrombosis in all the lower limbs of 40 cases. Among them, 24 cases were received total hip arthroplasties, 4 cases were received total knee arthroplasties and 12 cases were received dynamic hip screw. Postoperatively, every case were continuously received intermittent pneumatic compression for 14 to 21 days (2 hours, qid), and venography were performed on the operated lower limb on the 7th day after operation to check the presence of deep venous thrombosis. RESULTS: Among the 40 cases, there were 4 cases of deep venous thrombosis without symptom of pulmonary embolism, the incidence rate was 10%. CONCLUSION: Intermittent pneumatic compression can significantly reduce the incidence rate of deep venous thrombosis after the operation of the lower limbs. PMID- 12080846 TI - [Longitudinal biomechanical effect on repair options of artery injury]. AB - OBJECTIVE: To investigate the relationship between the different defect length of vessels and the options of vascular repair, and to compare the different options of repair because of the longitudinal biomechanical effect. METHODS: A clinical analysis was undertaken to evaluate the major arterial and venous injuries in human extremities repaired by end-to-end anastomoses or venous autograft(177 cases, 185 vessels). Compared the defect length of the same kind of vessels repaired by different options (Student-t test). Evaluated the defect length to repair arterial injuries between by end-to-end anastomoses and by vein graft by means of 95% confidence interval. RESULTS: There was significant difference between the defect length of brachial artery repaired by end-to-end anastomosis and femoral artery and popliteal artery repaired by autogenous vein graft (P < 0.01). The upper limit of confidence interval in the defect length of brachial artery, femoral artery and popliteal artery was 3.17 cm, 2.81 cm and 2.44 cm respectively by end-to-end anastomosis by means of 95% confidence interval. The lower limit of confidence interval in the defect length of brachial artery, femoral artery and popliteal artery was 2.82 cm, 2.41 cm and 2.17 cm respectively by vein graft by means of 95% confidence interval. The defect length of brachial artery, femoral artery and popliteal artery repaired by vein graft was linear correlation with the length of graft. CONCLUSION: Because of the longitudinal biomechanical difference of arteries and veins in human extremities, different options of repair are necessary to different arterial injuries. PMID- 12080847 TI - [Experimental study on the extent of injured blood vessels in an avulsion amputation model]. AB - OBJECTIVE: To measure the length and extent of the injured blood vessels in an avulsion amputation model. METHODS: Twenty rabbits were randomly divided into 2 groups. Group A was a sharp amputation group, and group B was an avulsion amputation group. The length and extent of the injured blood vessel was observed with naked eye, operation microscope and electron microscope, and the limbs were replanted. Group A and B were explored at three days and ten days after the replantation respectively. The patency rate and healing process were compared. RESULTS: All the severed ends of vessels in group A were neat with almost the same injured range in the three layers of the vessel wall about 1 mm away from the severed end. The vessels of group B were damaged seriously, the endothelial cells were deleted. The "jumping-like" damage could be observed in the elastic fibers. The injury of 2 to 3 mm away from the normal vessel wall could be observed by operation microscope. CONCLUSION: The damage of avulsion amputation vessels was irregular, 2 to 3 mm or more tissues should be excised under the microscope in the process of operation in order to ensure the healthy intact blood vessel walls. PMID- 12080848 TI - [Experimental study of mechanism and measures of prevention and treatment of hypothermal vasoconstriction]. AB - OBJECTIVE: To investigate the mechanism and to explore the measures of prevention and treatment of hypothermal vasoconstriction. METHODS: By the techniques of endothelial cell culture and scanning electron microscopy, and vasomotor functional test of isolated vascular vessels, the relation of hypothermal vasoconstriction and the release of endothelium-derived contractile and vasodilative factors were observed. RESULTS: Hypothermia obviously induced vasoconstriction of isolated vascular vessels, whether endothelium was intact or removed, the lower the temperature, the higher the vascular tension. Removal of endothelium could decrease the effect of vasoconstriction by hypothermia. The conditioned medium of bovine aortic endothelial cell could induce significantly vasoconstriction of isolated rat common neck arterial ring in hypothermia. It indicated that the bovine aortic endothelial cells secreted contractile factors into the medium. Reheating to 37 degrees C or vasodilator or reheating plus vasodilator did not obviously influence the hypothermia-induced vasoconstriction within 2 hours. When reheating to 50 degrees C, vascular tension was decreased, but only changed in range of 28% to 42%. CONCLUSION: Hypothermia vasoconstriction is relative to vasoconstrictor factors secreted by endothelium. Reheating to 37 degrees C or vasodilator does not antagonize the constriction of vascular vessels. Reheating to 50 degrees C only partially eliminates the constrict effect of blood vessels, so the prevention of hypothermia vasoconstriction should be emphasized. PMID- 12080849 TI - [Property of compliance and change of structural components in anastomosed artery]. AB - OBJECTIVE: To explore the relationship between the properties of compliance and the change of structure of components in anastomosed arteries. METHODS: The arterial pressure and diameter of femoral arteries of dogs were measured in vivo before and after arterial anastomosis in different time intervals to deduce the arterial compliance. The anastomosed arteries were removed and evaluated through light microscopic examination and various staining methods, the relative contents of elastin, collagen and smooth muscles were measured through image analysis system. RESULTS: The compliance of arteries was gradually decreased after anastomosis with peak-time on the 14th day. The content of elastin at different time had no significant difference, while the content of collagen increased gradually, the ratio of them was increased. CONCLUSION: The property of compliance of anastomosed arteries is closely related to the contents of the structural components. PMID- 12080851 TI - [Changes of the responses of arterioles to noradrenaline, acetylcholine and nitroglycerin after limb ischemia-reperfusion]. AB - OBJECTIVE: To explore the mechanism of microvascular spasm after limb ischemia reperfusion. METHODS: The rabbit hindlimb normothermic tourniquet ischemia model was employed. The tendon on the dorsum of the foot was exposed for observation of microvessels. The responses of arterioles on tendon surface to topical application of 10(-6) mol/L noradrenaline (NE) (a vasoconstrictor), 10(-6) mol/l acetylcholine(Ach) (an endothelium-dependent vasodilator) and 10(-4) mol/L nitroglycerin(NTG) (an endothelium-independent vasodilator) were observed at the period of ischemia and following 30 minutes of reperfusion after 2 hours and 5 hours of ischemia by use of intravital microscopy. RESULTS: No significant changes in the responses of arterioles to NE, Ach and NTG were noted following 30 minutes of reperfusion after 2 hours of ischemia compared with pre-ischemia. The constrictor responses of arterioles to NE were still not significantly altered following 30 minutes of reperfusion after 5 hours of ischemia, however, the dilation responses to Ach and NTG were significantly decreased (to Ach P < 0.01; to NTG, P < 0.05). CONCLUSION: Reperfusion after 5 hours of ischemia significantly impairs both the endothelium-dependent and endothelium-independent vasodilation, meanwhile preserves constrictor responses to NE, these may contribute to the genesis of the vasospasm in ischemia reperfusion. PMID- 12080850 TI - [Experimental evaluation of low-dose aspirin used in microvascular surgery]. AB - OBJECTIVE: To evaluate the effect of low-dose aspirin on the deposition of platelet at the anastomotic site and the function of coagulation system in order to provide experimental data for clinical use. METHODS: (1) Twenty-eight SD rats were divided into experimental group (n = 21) and control group (n = 7), aspirin were administered through a catheter placed in the femoral vein in dose of 4 mg/kg in the experimental group and the same dose of normal saline in the control group. The experimental group was subdivided into 3 groups, with 7 rats in each group, according to survival time of 24, 48 and 72 hours after dose. Samples of 4 ml blood were taken by heart puncture from each rat to investigate the maximal platelet aggregation rate(MAR), prothrombin time(PT) and kaolin partial thromboplastin time(KPTT). (2) Sixteen New Zealand White rabbits were divided into experimental and control group, 8 rabbits in each group. Drugs were given in the same way. Forty-eight hours later, the bilateral femoral arteries of each rabbit were exposed and arteries between inguinal ligament and the origin of the superficial epigastric arteries were transected and end-to-end anastomosis was completed with interrupted suturing technique. Fifteen and 120 minutes after the recovery of blood flow, the left and the right vessels containing anastomotic sites were harvested respectively and treated with 125I-labeled anti-GP IIb/III a antibody (SZ-21) using radioimmunobinding method. The radioactivities of the anastomosed vessels were measured. RESULTS: The KPTT in the experimental group was longer than that of the control group at 24- and 48-hour group, the mean percentages of increase were 42.56% and 35.33% respectively, and there were very significant differences between the experimental and control group in 24-hour group (P < 0.001). The PT value in experimental group was longer than that of the control group, but there was no significant difference (P > 0.05), and the maximal aggregation rate of platelet in the experimental group was significantly lower than that of the control group after 72 hours (P < 0.001). The radioactivity of the anastomosed arteries in the experimental group were significantly higher than that of the control group (P < 0.001) at 15 minutes after the recovery of blood flow, the mean percentage of increase was 110%. CONCLUSION: Low-dose aspirin can significantly affect the function of the intrinsic coagulation system, prevent the aggregation of platelets, but no effect on the function of the extrinsic coagulation system. On the other hand, it can also increase the deposition of platelet on the anastomotic sites after end-to end anastomosis, especially in the early stage when it is intravenously injected, but it is strong enough to cause thrombosis at the anastomotic sites. The effects of low dose aspirin on the coagulation system are inconsistent with its local effects on anastomotic sites. PMID- 12080852 TI - [Transposition of muscular skeletal flap pedicled with straight head of rectus femoris for treatment of avascular necrosis of femoral head]. AB - OBJECTIVE: To investigate the clinical results of transposition of muscular skeletal flap pedicled with straight head of rectus femoris for treatment of avascular necrosis of adult femoral head. METHODS: Eight patients with avascular necrosis of femoral head were adopted in this study. There were 6 males and 2 females, the ages were ranged from 24 to 56 years. According to the criteria of Ficat, there were 5 cases in stage II and 3 cases in stage III. The Smith Peterson incision was used to expose the capsule of the hip. After complete curettage of the necrotic bone from the femoral head, the muscular skeletal flap pedicled with straight head of rectus femoris was resected and transposited into femoral head. Finally, conventional decompression of head was performed. RESULTS: All the cases were followed up for 1 to 3 years. There were excellent results in 5 cases, good in 2 cases and moderate in 1 case. The rate of excellent and good results were 87.5%. CONCLUSION: Comparing with other pedicled bony flaps, the muscular skeletal flap pedicled with straight head of rectus femoris is characterized by its convenience and efficacy. It is suitable for the treatment of avascular necrosis of femoral head in stage II or III, but the contour of the femoral head should be nearly normal. PMID- 12080853 TI - [Treatment of tibia and femur massive defect with pedicled fibula transposition]. AB - OBJECTIVE: To investigate the repairing result for the massive bony defects of upper and middle tibia and lower femur. METHODS: Since 1974, four types of pedicled-fibula transposition were performed to repair the massive bone defect of tibia and femur in 25 cases, which included; 9 cases with benign tumor of upper part of tibia were performed muscle-pedicled fibula transposition and knee fusion after tumor resection; 9 cases with extensive benign tumor or tumoroid lesion of tibia shaft were performed muscle-pedicled fibula transposition and tibia-fibula fusion after tumor resection; 2 cases with extensive benign tumor or tumoroid lesion of middle and lower parts of tibia were performed vascular pedicled fibula transposition and tibia-fibula fusion; 5 cases with benign tumor of distal femur were performed vascular pedicled fibula reversal transposition and knee fusion. RESULTS: After 3 months to 11 years follow-up, 23 cases showed bone healing at 6 months postoperatively. The other 2 cases showed bone healing at 12 months postoperatively. All cases had satisfactory functional rehabilitation. CONCLUSION: Pedicled-fibula transposition is a choice method for repairing massive defects of tibia and femur. PMID- 12080854 TI - [Bone dynamics of repair of mandibular defect with collagen/hydroxyapatite]. AB - OBJECTIVE: To study the effect of collagen/hydroxyapatite(CHA) instead of autogenous bone transplantation on repairing the mandibular defects. METHODS: Ten Chinese experimental minipigs were made 2 cm bone defects in diameter in the mandible. The experimental group was implanted CHA, while the control group was implanted autogenous bone. The basic parameters of bone dynamics were determined by bone metrology. RESULTS: There was remarkable difference between the two groups in the mean distance and mineralization apposition rate of double label bands marked by tetracycline(P < 0.05), while the mean osteoid seam width and mineralization lag time had no remarkable difference(P > 0.05). It suggested that CHA had good osteogenesis. The collagen in CHA offered the condition of bone mineralization, and the mineralization peak of experimental group was present at 4 weeks earlier than that of control group (8 weeks). CONCLUSION: CHA may be a substitute of autogenous bone transplantation in repairing the mandibular defects, and the second operation for offering the implanting bone is avoidable, therefore, CHA may be an ideal material to repair bone defects. PMID- 12080856 TI - [Comparison of several rib rings with intercostal muscles for tracheal replacement in dogs]. AB - OBJECTIVE: To compare the effect of several types of rib rings with intercostal muscles for the replacement of trachea in thorax. METHODS: The surface layer of the third rib of dogs were ripped off and curved into triangular, quadrilateral and polygonal form. These three types of rib rings with intercostal muscles were used to replace a segment of trachea in thorax. RESULTS: The stability of triangular rib ring was very well, but stricture of ring were often happened because of its smaller internal diameter. These stability of quadrilateral rib ring was the worst. The polygonal rib ring presented the biggest diameter and good stability compared to the other two kinds of rings. If silicone tube was supplemented in the polygonal rib ring, the quality of artificial trachea was excellent. CONCLUSION: The rib rings with intercostal muscles are successfully used for replacing the defect of trachea in canine thorax. The polygonal rib rings have the best quality in the three types of rib ring for tracheal replacement. PMID- 12080855 TI - [The effect of cefazolin loaded bone matrix gelatin on repairing large segmental bone defects and preventing infection after operation]. AB - OBJECTIVE: To explore the possibility of repair long segmental bone defects and preventing infection with cefazolin loaded bone matrix gelatin (C-BMG). METHODS: C-BMG was made from putting cefazolin into BMG by vacuum adsorption and freeze drying techniques. The sustaining period of effective drug concentration in vitro and in vivo was detected by inhabition bacteria, and the drug concentration in local tissues (bone and muscle) and plasma after implantation of C-BMG was examined by high performance liquid chromatography(HPLC). RESULTS: The effective inhibition time to staphylococcus aureus of C-BMG was 22 days in vitro, while 14 days in vivo. The drug concentration in local tissues(bone and muscle) were higher than that of plasma, and the drug concentration in local tissues was higher in early stage, later it kept stable low drug release. It suggested that C BMG had excellent ability to repair segmental long bone defects. CONCLUSION: C BMG can gradually release cefazolin with effective drug concentration and has excellent ability to repair segmental long bone defects. It may be a novel method to repair segmental long bone defects and prevent infection after the operation. PMID- 12080857 TI - [SV40 and cell immortalization]. AB - OBJECTIVE: To explore the SV40-mediated immortalization, the related factors and their roles in cell immortalization. METHODS: The original articles about cell immortalization and replicative senescence in recent decade were reviewed. RESULTS: Cell immortalization was a multifaceted phenomenon, it was involved in viral DNA integration, activation of telomerase, inactivation of growth suppressors, and so on, and their roles were closely related. CONCLUSION: The research on cell immortalization may be expected to provide important insights into a broad range of cellular biological phenomenon, and the immortalized cells can play important roles in the research of cell engineering and tissue engineering as standard cells. PMID- 12080858 TI - [Development of biodegradable polymer scaffolds for bone tissue engineering]. AB - OBJECTIVE: To investigate the selection and manufacture of ideal extracellular matrix materials in bone tissue engineering. METHODS: The recent literatures about biodegradable polymers served as culture scaffolds of osteoblasts were widely reviewed, the advantages and disadvantages of biodegradable synthetic polymers and natural polymers were analysed. RESULTS: The ideal extracellular matrix material in bone tissue engineering should be made up of inorganic materials, synthetic polymers and natural polymers, which possesses morphological structure of three-dimensional foam with self-mediated drug slow delivery system of bone growth factors. CONCLUSION: The design and manufacture of combined extracellular matrix materials in bone tissue engineering is a very important and urgent challenge. PMID- 12080859 TI - [Xenogeneic graft-versus-host disease]. AB - OBJECTIVE: To evaluate the histological characteristics of xenogeneic graft versus-host disease (GVHD), and to differentiate it from the allogeneic GVHD. METHODS: The original articles related to the characteristics of xenogeneic GVHD and the relationship between xenogeneic GVHD and allogeneic GVHD were widely reviewed in the past decade. RESULTS: Xenogeneic GVHD was different from allogeneic GVHD in the speed of development, position and features of pathological changes, due to the incompatibility of common simulative signals, adhesive molecules and difference of antigens. Xenogeneic spleen transplantation could induce xenogeneic GVHD. CONCLUSION: Xenogeneic GVHD can be used as a model to mimic rejection in xenotransplantation and to evaluate the compatibility of xenotransplantation. PMID- 12080860 TI - [Research progress on preservation of severed limbs]. AB - OBJECTIVE: To define how to preserve the severed limbs to prolong the period of replantation. METHODS: The original articles about preservation of severed limbs in recent years were reviewed, it was suggested that the period of replantation was determined by the injury of skeletal muscle. RESULTS: When the environment of severed limbs was changed, the injures of skeletal muscle could be decreased. CONCLUSION: After the severed limbs are reasonably preserved, the period of replantation may be prolonged. PMID- 12080861 TI - Health care systems may offer 'open health management'. PMID- 12080862 TI - Greater nursing staff levels result in better care for hospital patients. PMID- 12080863 TI - Hospitals must raise rates, make profits. PMID- 12080864 TI - Reaching out. PMID- 12080865 TI - To know what sort of person has a disease. PMID- 12080866 TI - Leaving a legacy. PMID- 12080867 TI - A story of Pick's disease: a rare form of dementia. AB - Pick's disease is a progressive illness that affects brain function, eventually causing loss of verbal skills and problem-solving abilities. Pick's disease accounts for 5% of all dementias. The most common symptoms involve personality, behavior, and language changes. Diagnosis can be difficult; a differential diagnosis often requires several years. Frequently, the disorder is initially diagnosed as stress, depression, or Alzheimer's disease. A magnetic resonance imaging scan will show atrophied portions of the frontal and temporal lobes. One of the most difficult aspects of care for someone with Pick's disease is communication. The communication styles of nurses can alleviate the anxiety experienced by a person with Pick's disease. Nurses should try different strategies as an approach to communicating and caring for someone with Pick's disease, remembering that each person suffering from dementia is different. Pick's disease can be emotionally devastating to the families of those who develop this disease. Working with families confronting the disability of a loved one is perhaps the greatest challenge for nurses. Nurses have the responsibility of educating the primary caregiver about nutrition, skin protection, incontinence care, safety, and end-of-life decisions. PMID- 12080868 TI - Addressing spasticity-related pain in children with spastic cerebral palsy. AB - Children with spastic cerebral palsy (CP) often suffer pain as a result of their spasms. Some studies have suggested that the pain experienced by these children may be poorly controlled or respond poorly to treatment. Limited research is available that addresses the treatment of pain due to spasms in children. Therefore, clinicians do not have empirical evidence to guide their decisions. Many current treatment options have not been tested for their effect on pain. Previous research has been guided more by the ability of these treatments to increase functional abilities. Pain also is an important indicator of quality of life; however, this has been overlooked in research. PMID- 12080870 TI - Reproductive decision making in women with multiple sclerosis. AB - Multiple sclerosis (MS), the most common acquired neurological disorder of young adults, often strikes young women in their childbearing years. Despite the overlap of MS onset with the childbearing years of women, little is known about how women with MS make decisions about pregnancy and childbearing. In an effort to understand the process of decision making in these women, an exploratory descriptive qualitative study was undertaken; 15 pregnant women with MS were interviewed about their decisions to become pregnant and the factors that entered into their decision-making processes. Content analysis was used to identify and describe the fears and concerns of women with MS around the process of making decisions and about how the presence of MS had influenced that process. The unpredictability of MS and the effect that pregnancy might have on MS was a strong theme that emerged from the analysis. Although participants in this study had proceeded to become pregnant despite their fears related to the effect of pregnancy on the course of their MS, they continued to perceive their decision as risky. The diagnosis of MS affected their previous plans for number of children as well as spacing of pregnancies. PMID- 12080869 TI - The impact of aneurysmal subarachnoid hemorrhage on functional outcome. AB - Despite advances in the management of aneurysmal subarachnoid hemorrhage (SAH), a significant percentage of survivors are left with persistent cognitive, behavioral, and emotional changes that affect their day-to-day lives. This article describes outcome at 3 months after aneurysmal SAH in 61 patients, using the Extended Glasgow Outcome Scale (GOSE) and the Functional Status Examination (FSE). The GOSE provides a measure of overall functional outcome but does not address the specifics of functional limitations. The FSE, in addition to identifying functional limitations, provides insight into factors contributing to them and the extent to which SAH survivors perceive them as affecting their day to-day activities. The findings of this study demonstrate that SAH survivors have considerable limitations in functional status in almost all areas of daily living at 3 months following SAH. The limitations were attributed to a variety of physical, cognitive, and emotional factors, and they were reported to be moderately to severely bothersome in almost half of the individuals. The findings highlight the need for appropriate rehabilitation, education, and support for SAH survivors and their families to enhance coping and improve quality of life, given the substantial and persistent impact of SAH. PMID- 12080871 TI - Kernohan's notch phenomenon: a case study. AB - In a patient suffering from brain herniation due to a right-sided subdural hematoma, a neurological examination should show left-sided deficits, known as localizing signs, and a decreased level of consciousness. However, false localizing signs may be present, attributed to pressure on Kernohan's notch. A case study demonstrates these false localizing signs, known as Kernohan's notch phenomenon. PMID- 12080872 TI - Pharmacology update. Dietary supplements: applying the knowledge. PMID- 12080873 TI - [Clinical and molecular study of Chilean patients with McCune-Albright syndrome]. AB - BACKGROUND: McCune-Albright Syndrome (MAS) is characterized by precocious puberty, "cafe au lait" skin lesions and polyostotic fibrous dysplasia. It is caused by 4 post-zygotic mutations of G alpha s protein with a mosaic distribution. AIM: To describe the clinical presentation and to investigate the presence of the Arg by his substitution (R201H) in 14 girls with MAS. PATIENTS AND METHODS: We performed a clinical analysis of the patients and specific allele PCR in DNA obtained from leukocytes. RESULTS: Twelve of 14 patients presented with precocious puberty, one with cyclical vaginal bleeding and one with pathological bone fractures. Eight girls had polyostotic fibrous dysplasia, one had hyperthyroidism, four had pathological fractures, ten had ovarian cysts, six had breast hyperpigmentation and ten had "cafe au lait" skin lesions. We detected the R2O1H mutation in 10 of 14 patients. We found no difference in the severity of symptoms or in the age of presentation between the patients with and without the mutation. CONCLUSIONS: The R201H mutation can be detected in white blood cells, in approximately 70% of cases. Patients exhibit wide clinical variability with the same molecular defect. This suggests that tissues have different proportions of mutant cells. PMID- 12080874 TI - [heparin and insulin treatment of acute pancreatitis caused by hypertriglyceridemia. Experience of 5 cases]. AB - BACKGROUND: Hypertriglyceridemia over 1,000 mg/dl can provoke acute pancreatitis and its persistence can worsen the clinical outcome. On the contrary, a rapid decrease in triglyceride level is beneficial. Plasmapheresis has been performed in some patients to remove chylomicrons from the circulation, while heparin and/or insulin have been administered in some other cases to rapidly reduce blood triglycerides. Heparin and insulin stimulate lipoprotein-lipase activity and accelerate chylomicron degradation. AIM: To report five patients with acute pancreatitis treated with heparin and insulin. PATIENTS AND METHODS: Five patients (4 females and 1 male) seen in the last two years, who suffered acute pancreatitis induced by hypertriglyceridemia are reported. Initial blood triglyceride levels were above 1,000 mg/dl (range 1,590-8,690 mg/dl). Besides the usual treatment of acute pancreatitis, heparin and/or insulin were administered intravenously in continuous infusion. Heparin dose was guided by usual parameters of blood coagulation, and insulin dose, by serial determinations of blood glucose. Pancreatic necrosis was demonstrated in 4 patients. RESULTS: Serum triglyceride levels decreased to < 500 mg/dl within 3 days in all cases. No complication of treatment was observed and all patients survived. Early and late complications of pancreatitis occurred in one patient. CONCLUSION: Administration of heparin and/or insulin is an efficient alternative to reduce triglyceride levels in patients with acute pancreatitis and hypertriglyceridemia. PMID- 12080875 TI - [Is the epidemiological surveillance system of nosocomial infections recommended by the American CDC applicable in a Chilean hospital?]. AB - BACKGROUND: The National Nosocomial Infections Surveillance System (NNIS system) is the method for surveying nosocomial infections used by the Centers for Disease Control (CDC). This strategy allows the comparison of different hospitals, using rate adjustments. In Chile, this system is not used. AIM: To report the application of this system in a tertiary reference hospital in Chile. MATERIALS AND METHODS: We performed a six months prospective cohort study. The active surveillance was carried out by using the intensive care unit and surgery components of the NNIS system. Tabulation and analysis of the data were done according to the NNIS system. In a parallel prevalence study, we determined the NNIS system sensitivity to detect nosocomial infections. RESULTS: A total of 492 patients were followed with a global nosocomial infection rate of 14%, for discharged patients. The calculated sensitivity and specificity of the system was 84.2 and 97% respectively. In the intensive care unit, 45 of 169 patients had nosocomial infections, with an adjusted rate of 2.8% for mean hospitalization time and severity of illness. In the cardiovascular and thoracic surgical units, 216 and 107 procedures were surveyed, respectively. The global rates of nosocomial infections were 7.4 and 7.5%, respectively. The adjusted rates according to risk factors were 0.9 and 2.3%, respectively. CONCLUSIONS: These data indicate that the surgical units had surgical site infections rates similar to those reported by the CDC. Nosocomial infections rates in Chile can be compared with rates observed in other countries. The epidemiological data collected can be useful to focus intervention or preventive strategies. PMID- 12080876 TI - [Endoscopic ligation plus sclerotherapy in esophageal varices treatment. Experience in active bleeding and in the elective treatment]. AB - BACKGROUND: Endoscopic ligation is the treatment of choice for bleeding esophageal varices. The usefulness of additional sclerotherapy is not clear. AIM: To assess the effectiveness of ligation followed by sclerotherapy in the treatment of variceal bleeding. PATIENTS AND METHODS: Forty eight patients with variceal bleeding admitted for emergency treatment and 73 patients admitted for elective treatment were studied. Varices were ligated until a significant reduction in size was achieved. Eradication was completed with the injection of 1% polidocanol. RESULTS: In 34 of 48 patients admitted for emergency treatment, the site of variceal rupture was identified. In all these subjects, and in 13 of 14 patients in whom the rupture site was not identified, hemorrhage was stopped with the procedure. Varices were eradicated in 108 of the 121 patients. Hemorrhage recidivated in 12.5% of patients admitted for emergency treatment, in a period of 14 months of follow up and in 9.6% of those admitted for elective treatment, in a period of 16 months follow up. Mortality was 14, 18 and 57% among patients classified as Child Pugh A, B or C, respectively. CONCLUSIONS: Endoscopic ligation is effective in the treatment of variceal bleeding. Adding sclerotherapy, variceal eradication is achieved in a high percentage of patients. Survival depends on hepatic function. PMID- 12080877 TI - [Comparative study of health care costs for patients using home oxygen therapy or those in waiting lists]. AB - BACKGROUND: Home oxygen therapy is the only intervention that significantly prolongs survival of patients with chronic obstructive pulmonary disease. Since this therapy is expensive, it is imperative to calculate its cost-benefit ratio. AIM: To compare health care costs for patients in a home oxygen therapy program, with those of a similar group of patients in a waiting list for this therapy. MATERIAL AND METHODS: A retrospective analysis of 21 patients on home oxygen therapy for 36 months or less and a group of 13 patients in the waiting list during a similar lapse. Medical consultations, drugs delivered and the cost of oxygen therapy were considered among ambulatory costs. Hospitalization costs included the number of days on regular or intermediary care beds, laboratory tests and drugs delivered. RESULTS: The annual health care costs for patients on home oxygen therapy were 709,656 Chilean pesos and the costs for patients in the waiting list were 797,320 Chilean pesos. CONCLUSIONS: The overall health care costs of home oxygen therapy are similar to the cost required by patients with chronic obstructive pulmonary disease remaining in waiting lists. PMID- 12080879 TI - [Child abuse in in the city Temuco. Prevalence study and associated factors]. AB - BACKGROUND: Child maltreatment is recognised as a significant health problem in developed countries. There is increasing awareness on family violence in Chile, becoming a health priority in the last five years, but there is scant information about its prevalence. AIM: Determine prevalence of child abuse, and associated factors in a population sample from Temuco, Chile. MATERIAL AND METHODS: A standardised questionnaire was applied to a sample of 422 women at their homes. Instrument had questions about disciplinary practices by caretakers, a standardised questionnaire to assess mental health, as measures of risk and protective factors for violence RESULTS: The prevalence of psychological aggression delivered by mothers or fathers was 17.5% and 6.8% respectively. The figures for corporal punishment delivered by mother or fathers were 42.3% and 17% respectively. Three percent of mothers and 1.2% of fathers recognised severe physical abuse. Associated factors were mother impaired mental health, antecedents of child abuse in both parents, parents alcohol abuse and child emotional/behavioral problem. CONCLUSION: Child abuse is a extended significant problem in Temuco that requires multidisciplinary intervention programs. PMID- 12080878 TI - [Conjugal violence in the city Temuco. Prevalence studies and associated factors]. AB - BACKGROUND: Violence against women is a recognized public health problem in developed countries. There is increasing awareness on family violence in Chile, but there is scant information about its prevalence. AIM: To determine the prevalence of family violence against women in a population sample in Temuco, Chile. MATERIAL AND METHODS: A standardized questionnaire about family violence was applied to a sample of 422 women at their homes. The questionnaire had six sections that included a list of violent behaviors from husbands or partners, a standardized instrument to assess mental health and alcohol consumption by the couple, history of child abuse, community support, type of employment and years of school education. RESULTS: Forty nine percent of women reported psychological aggression, 13% reported physical violence and 5.5%, sexual violence. The presence of anxiety or depressive symptoms, being witness of violence between parents during childhood, a lower educational level, being a housewife, alcohol abuse and lack of community support were risk factors for violence among women. Among men, the history of child abuse, a lower education level, and alcohol abuse were identified as risk factors for violent behaviors. CONCLUSIONS: Conjugal violence is a significant mental health problem in Temuco, Chile. PMID- 12080881 TI - [Ruptured thoracic aorta aneurysm: endovascular treatment]. AB - In 1991, a technique to exclude aortic aneurysms from circulation inserting an endoluminal graft through the femoral artery, was described. This procedure, usually used for elective abdominal aneurysms, can also be used in the thoracic aorta. We report a 41 years old male with a Marfan syndrome, presenting with a descending aorta aneurysm that ruptured to the mediastinum and pleural cavity. He was compensated hemodynamically and an endovascular stent-graft was deployed at the ruptured zone, through the femoral artery. The postoperative evolution of the patient was uneventful. This technique will allow a less invasive treatment of ruptured aortic aneurysms. PMID- 12080880 TI - [Primary cholangiocarcinoma associated with Caroli disease]. AB - A 68 years old male presented with right hypochondrium abdominal pain and jaundice with no other clinical finding. CAT Scan and Ultrasonography showed right lobe bile duct dilatation. Magnetic Resonance Cholangiopancreatography gave an outstanding vision of a restricted right lobe bile duct dilatation associated to choledocholithiasis. A right hepatectomy and bile duct exploration were performed. The histopathological study disclosed a Caroli disease associated to a primary cholangiocarcinoma. Caroli disease is a congenital disorder characterized by intrahepatic cystic bile duct dilatation with a high risk association with cholangiocarcinoma. PMID- 12080882 TI - [Spinal cord metastasis: an uncommon location. Experience in 2 cases]. AB - Spinal cord metastases are an uncommon secondary location of a malignant neoplasm. They are rarely diagnosed during life and when that is the case, it is in the clinical setting of a disseminated cancer and very seldom as the first clinical manifestation. We report two patients, with no previous disease, who developed a progressive myelopathy. An intramedullary spinal cord tumor was diagnosed, based on the clinical picture and imaging studies. They were operated and biopsies showed spinal cord metastases whose primary tumor was a lung neoplasm. We discuss the clinical features in these patients, the diagnosis of progressive myelopathy in cancer patients, treatment and prognosis of this unusual secondary cancer location. PMID- 12080883 TI - [Febrile neutropenia in the child with cancer. Current concepts about risk assessment and selective management]. AB - The risk for invasive bacterial infection (IBI) in cancer pediatric patients with febrile neutropenia, is variable. Clinicians worldwide are increasingly considering selective strategies for children at low risk for IBI, including shortened antimicrobial course, early hospital discharge, oral antimicrobial treatment, and management as outpatients. These strategies would significantly benefit these children and health care systems. The critical issue is to identify the most reliable risk factors useful for selection of those individuals who are at low risk for IBI. In Chile, during the past 10 years, a group of physicians from the Subcommittee of Infectious Diseases of the National Child Program of Antineoplastic Drugs and the University of Chile have worked to develop more selective strategies for pediatric patients with cancer, fever and neutropenia. During 1996-1997 we identified risk factors of IBI in a group of 447 febrile neutropenic episodes. During 1999-2000 we validated these risk factors in a prospective study that included 263 febrile neutropenic episodes. A model of risk prediction was developed and is currently being evaluated for the selection of low risk patients who are treated as inpatients for 24 hours, followed by outpatient treatment. PMID- 12080884 TI - [Medical errors]. AB - Untoward effects can result from the various procedures involved in patients care. Because of its risks and consequences it is very important to determine the causal factors of these events to improve the procedures at stake and be sure to obtain from them the results to be expected. Several recent and somehow disturbing reports about the magnitude and type of these said medical errors, have raised interest to review the subject and phase this problem under the scope of a quality control system, as to prevent its occurrence or mitigate its effects when there is no chance to avoid them properly. PMID- 12080885 TI - [Sources of error in clinical practice]. AB - The clinical setting is everything that contact us with the patient's reality, including physical, instrumental and laboratory exploration, and we use it for diagnostic and treatment purposes. Diagnosis is the synthesis by which physicians conclude an elaborated analysis of subjective and objective data, obtained by physical exam and technology. This procedure involves data collection about the patient and his disease, the results of various tests and laboratory findings, the interpretation of the data and a diagnostic summary. Medical errors may be derived from insufficiency in any of these phases. It is evident that technology will never replace the intellectual data recollection, analysis and reasoning that is the physician's art. This contribution underlines some of the defects frequently observed in clinical practice that may cause medical errors. PMID- 12080886 TI - [Ethical reflections on errors in medicine]. AB - It is usually inevitable to save the physician-patient relationship from the effects of medical errors. It is so because either the professional responsibility, the patients trust and the principles that govern the medical action are compromised by such events. The possibility to learn from it, prevent its appearance, improve the procedures and curtail its consequences, is linked to developing ethical attitudes to overcome these circumstances without further legal accountability. PMID- 12080887 TI - [Definitions regarding medical errors]. AB - Medical errors can be defined as value laden facts, in relation to guidelines or expectations, to the responsibility involved, to the obligations abated, to the pertinence of excuses afforded because of complexity, compliance and prudence or its aggravation through recklessness. Due care is related to prevention and corresponding attitudes. Professionalism, continuous education and quality control systems to detect errors without dispensing inexcusable faults, are the main tools to avoid medical errors. PMID- 12080888 TI - [Management of errors in medicine]. AB - If negligent actions are ruled out, medical errors must be prevented or attenuated enhancing security in all the health care system. Prudent and confidential information and assessment are crucial to obtain good results, discarding to look for whom to blame and assuming collectively the supervision of suspected conditions. Massive introduction of technology, intervention of multiple agents in the care of patients and the number of alternatives offered to manage health problems, contribute to increase medical errors. Not of lesser importance are the increasing expectations of patients on health benefits, that must be synchronized to the continuous improvement in medical care. PMID- 12080889 TI - [Planning medical education for the next decades]. AB - Accelerated changes in source and contents of knowledge, methodologies and scenarios used in learning, and in the meaning and goals of the educational process, are typical of these times. To characterize future needs, roles and expectations of the various actors involved, is also very unreliable and unpredictable. Performing reiterative strategic analysis, monitoring procedures, doing continuous assessment of results and making permanent adaptations in the educational process to the environmental conditions, are unavoidable means to get secure and pertinent effects from the implemented actions. From this perspective, the University of Chile Faculty of Medicine is formulating its proposals to innovate in curriculum, and strengthen its academic resources and students development. PMID- 12080890 TI - [HIV blood test and informed consent]. PMID- 12080891 TI - An occupational health nursing education program. Relevance to nurses in nonoccupational practice settings. AB - A collaborative effort between community health faculty in an upper division nursing program and nurses from a nearby agricultural health and research center resulted in an educational program focused on occupational health and safety issues in the agricultural industry. The 3 hour class was presented each semester between 1997 and 1999 to RN students enrolled in a community health organization class. In addition to information about the health and safety hazards in agriculture, the nursing students learned about the center's research projects and clinical services available to the farming community. A follow up survey was mailed to the nursing students to evaluate the effectiveness and relevance of the occupational health program by identifying the proportion of the nurses' caseloads spent on occupational health problems, and whether the information and materials presented in the class were relevant to practice. Results showed that even though the surveyed nurses were employed in nonoccupational practice settings, nearly everyone rated the occupational health information as useful. Findings also revealed that although nurses in nonoccupational practice can spend nearly a quarter of their time caring for clients with work related health problems, they may lack adequate educational preparation to do so. PMID- 12080892 TI - Certified occupational health nurse specialists practicing as nurse practitioners. Administrative practices in the California workers' compensation system. PMID- 12080893 TI - Primary prevention of agricultural injuries. Use of standardized nursing diagnosis, interventions, and outcomes. AB - 1. The use of standardized nursing language is important to the advancement of nursing knowledge and practice. The standardization of nursing diagnoses, interventions, and outcomes provides a means for collecting and analyzing large clinical databases. 2. The occupational health nurse has a clear role in developing health and safety programs. Standardized language using North American Nursing Diagnosis Association diagnoses, nursing outcomes classification, and nursing intervention classification can be incorporated into these programs. 3. The use of standardized taxonomies in the primary prevention of cumulative trauma disorders demonstrates how these systems are integrated into the documentation of the nursing process in a nursing care plan. PMID- 12080894 TI - Improving return to work outcomes. Formalizing the process. AB - Potential for decreasing absences, saving disability costs, and improving productivity are important motivators for convincing employers to formalize their RTW program. The first step in formalizing a RTW program is developing a written corporate RTW policy statement. Another essential step is reviewing jobs and completing job analyses quantifying the physical and environmental requirements of the position. These valuable job analysis tools can be used to assist employers and health care providers with proactive transitional work options. Occupational health nurses must focus their skills on assessing, monitoring, and evaluating improvements in RTW outcomes, as well as translating their interventions into measurable value added services and cost savings. PMID- 12080895 TI - An approach for effective group teaching. PMID- 12080896 TI - Emergency contraception (post-coital Contraception). AB - Emergency Contraception is a post-coital contraceptive for women who have had unprotected intercourse or have reason to believe that their contraceptive method has failed. The article focuses mainly on Emergency Contraceptive Pills (ECPs) because they are the most frequently used form of post-coital contraception. In 1997 the FDA approved the "off-label" use of high dose of oral contraceptives for use as post-coital contraception. Since then, they have been approved for repackaging and marketed solely for use as post-coital emergency contraception. The first dose of ECPs must be administered within 72 hours of the act of unprotected intercourse. The second dose is taken 12 hours later. ECPs are believed to work in one of three ways depending on where the woman is in her menstrual cycle when she seeks treatment. It can delay or prevent ovulation, impair formation of the corpus luteum, or cause histological or biochemical changes within the endometrium, thus preventing implantation. Their effectiveness is approximately 75%, being most effective when administered as quickly as possible after the act of unprotected intercourse. The pills can cause nausea and vomiting, so the pre-administration of an anti-emetic may help alleviate these symptoms. A major issue concerning the ECPs is the lack of knowledge and availability. Very few health-care providers discuss ECPs with their patients. Most people cited the media as the primary source of information. The 72-hour window in which the ECPs must be administered makes it important for women to have easy access to these post-coital contraceptives. However, women who seek treatment will often find that their health-care provider will require a physical exam and/or a pregnancy test before writing a prescription. Yet, studies show that ECPs do not affect an implanted fetus, and there are no emergency contraceptive protocols that require pregnancy tests or physical exams prior to treatment. The AMA is encouraging physicians to better educate their patients about emergency contraceptives. Several health-care organizations are encouraging providers to supply women with an advance prescription for ECPs so that they will have immediate access to them, especially since most acts of unprotected intercourse occur at night or on weekends, when most clinics are closed. There is a possibility that ECPs might be available over-the-counter for women to have better access to the emergency contraceptive pills. PMID- 12080897 TI - Watershed events in the improvement of surgical outcomes in the elderly transplant (QLT) recipients. AB - The enactment of the Medicare legislation and the establishment of a National Institute for Aging in the 1960s and 1970s has spurred a number of developments which have proven to represent watershed advances in the surgical care of the elderly. The development of a multidisciplinary approach to both geriatric surgical care and research ultimately may prove to be the greatest advance yet seen. PMID- 12080898 TI - Case report: right-sided diverticulitis misdiagnosed as appendicitis both clinically and on plain film radiography. PMID- 12080899 TI - Prevention of intrauterine growth retardation. AB - Infant birth weight is the single most important factor affecting neonatal mortality. The risk factors are many, but several are preventable including smoking, alcohol use and drug abuse. The physician plays a vital role in both prevention and early detection of potential problems. PMID- 12080900 TI - No one is motivated to work any longer. PMID- 12080902 TI - Journal of the Oklahoma State Medical Association. PMID- 12080901 TI - Attitudes towards labor unions. PMID- 12080903 TI - Introduction to the special issue on repetitive behavior problems. PMID- 12080904 TI - Skin picking in German students. Prevalence, phenomenology, and associated characteristics. AB - Severe skin picking (SP) is a repetitive, intentionally performed behavior that causes noticeable tissue damage and results in clinically significant distress or impairment. To date, SP has received little attention in the psychiatric literature. This study was conducted to further investigate SP and its characteristics in a German student population. The participants (N = 133) completed various self-report questionnaires. More than 90% (n = 122) reported occasional SP, with six students (4.6%) endorsing significant impairment from recurrent, self-injurious SP. SP was triggered by specific cutaneous stimuli, situations, and emotions. The students primarily squeezed (85%) and scratched (77.4%) the skin, with a primary focus on the face (94.7%) and cuticles (52.6%). About 20% (n = 26) ate the picked tissue afterward. Results suggest SP is an underrecognized problem that occurs on a continuum ranging from mild to severe with generally stable clinical characteristics across cultures. PMID- 12080906 TI - Cognitive-behavior therapy for self-injurious skin picking. A case series. AB - Self-injurious skin picking is characterized by repetitive, ritualistic, or impulsive skin picking that leads to tissue damage and causes significant distress or impairment in daily functioning. Little is known about effective behavioral or cognitive-behavioral treatments for self-injurious skin picking. As described by Azrin and colleagues, habit reversal is a promising behavioral treatment for modifying nervous habits or tics. To the authors' knowledge, only one case series currently exists in the literature that shows self-injurious skin picking, in the absence of an underlying dermatological condition or without psychiatric comorbidity, can be successfully treated with habit reversal. In the current article, the authors describe the implementation and outcome of cognitive behavior therapy for three patients with severe self-injurious skin picking, two of which had psychiatric comorbidity. PMID- 12080905 TI - Body-focused repetitive behavior problems. Prevalence in a nonreferred population and differences in perceived somatic activity. AB - Body-focused repetitive behaviors (BFRBs) such as skin picking or scratching and nail biting can be physically and socially detrimental. Given the potential consequences associated with these behaviors, it is unfortunate this area has received relatively limited attention. The two purposes of the current study were (a) to determine the prevalence of BFRBs among typically developing persons and (b) to examine the contribution of reported somatic activity to the occurrence of BFRBs. Results indicated that 13.7% of the sample met criteria for at least one BFRB, of which the most common topography was nail biting. Persons with a BFRB reported significantly more somatic activity than persons without a BFRB. Further analyses revealed consistent findings across three separate topographies in which persons with a BFRB for nail biting, mouth chewing, and/or skin picking reported significantly more somatic activity than did persons without a BFRB. Clinical implications and diagnostic considerations are discussed. PMID- 12080907 TI - Acceptability of treatments for trichotillomania. Effects of age and severity. AB - Based on the literature, trichotillomania (TTM, chronic hair pulling) in children and adults appears to be responsive to behavioral interventions such as habit reversal. However, some have questioned the generality and acceptability of such procedures. This study compared the acceptability ratings of four interventions targeting TTM (habit reversal, hypnosis, medication, and punishment). In the study, 233 college students read case vignettes in which the age of the analogue client and the severity of the hair pulling were manipulated. Results showed significant differences between the four treatment conditions, with hypnosis and habit reversal being rated most acceptable. Age of the analogue client and severity of TTM did not significantly influence acceptability ratings. PMID- 12080908 TI - Treatment of early onset hair pulling as a simple habit. AB - The authors evaluated the effects of response prevention, a treatment previously shown to be effective for routine thumb sucking and suggested to be effective for early onset trichotillomania, applied to hair pulling in a 2-year-old. Response prevention was used alone in two settings (bedtime and naptime) and combined with a brief time out in another (daytime). The authors also used a novel assessment, weight of hairs pulled, and the results indicated complete cessation of hair pulling. Corresponding photographic evidence indicated complete regrowth of hair lost to pulling. These results add to a growing literature suggesting early onset hair pulling may be more appropriately classified as a benign habit than as trichotillomania. PMID- 12080909 TI - Sibling thumb sucking. Effects of treatment for targeted and untargeted siblings. AB - In this study, removal of a transitional object (pillow) was applied as the sole intervention for one of two siblings who sucked their thumbs. The intervention was applied only to the older sibling, whereas data were collected on the thumb sucking of both participants. Results indicated that removal of the transitional object for the older sibling reduced thumb sucking in both siblings and increased thumb sucking when the pillow was reintroduced. Discussion focuses on possible explanations for the effects of the intervention across both participants and the limitations of this study. PMID- 12080910 TI - A functional analysis of finger sucking in children. AB - In this investigation, the authors experimentally assessed the functions of finger sucking for 3 typically developing children ages 6, 7, and 14. In Experiment 1, a parent-conducted functional analysis, completed in each child's natural environment, showed that each of the children's finger sucking was most likely to occur when the child was alone, suggesting that the behaviors were maintained by automatic reinforcement. Experiment 2 involved assessing the nature of the sensory stimulation that maintained finger sucking by attenuating the sensory stimulation to the fingers via the use of Band-Aids and attenuating the sensory stimulation to the mouth with a mild numbing agent for 2 of the 3 children. For the 3rd child, mouthing objects were made available noncontingently to determine whether access to such items would result in low levels of finger sucking, suggesting reinforcer substitutability. Results of these analyses suggested that finger sucking was maintained by both oral and digital stimulation for 2 children and by oral stimulation for the 3rd. PMID- 12080911 TI - Paying for preparedness. PMID- 12080912 TI - Quality. Your results rewarded. AB - A new program is intended to sidestep the strained relationship between hospitals and managed care companies by awarding monetary grants to hospitals for their proven quality programs. PMID- 12080913 TI - Legal issues. After an assault. AB - In California, a health care worker who was attacked on the job by a patient sued her attacker. She won the lawsuit but lost her job. Hospitals are torn between obligations to people who come to them for care and employees who deliver it. PMID- 12080914 TI - Rural health care. Against the odds. PMID- 12080915 TI - Energy management. Hybrid heats up. AB - Managing energy costs can be a pain at best. But an Illinois hospital has found a way to take advantage of fluctuations in gas and electric prices by installing a hybrid HCAC system. PMID- 12080916 TI - HIPAA. Regs ramp up. PMID- 12080917 TI - International care. Crossing borders. AB - Searching for a new line of business to boost revenues, some providers look abroad. It's a decent strategy, but don't expect the big score right now: instead, keep looking at the long-term benefit. PMID- 12080919 TI - Tool time nurses. As handhelds proliferate, ease of use, logistics and the role of the device in direct patient care are keys to success. PMID- 12080918 TI - 1Q[3a]. How can we encourage more men to become nurses? PMID- 12080920 TI - . . . By a thread--a fragile, fraying safety net is everybody's problem. AB - With rising health insurance premiums, surging numbers of uninsured people, tighter state budgets and federal threats to cut back on Medicaid and other funding, the nation's already fragile safety net is unraveling. If you think that's a problem for public hospitals alone, think again. When the safety net frays, every hospital feels the pressure. PMID- 12080921 TI - CPOE (computerized physician order entry). An executive's guide. AB - Computerized physician order entry is controversial. Backers say it helps reduce medication errors significantly. Critics note that it's expensive, as well as complex to design and implement. This guide considers the advantages and costs of CPOE. PMID- 12080922 TI - NFP Investor Conference. Growth is back in sight. AB - In a whirlwind of high-stakes meetings in New York City in mid-May, two dozen of the nation's largest hospitals and health systems strutted their stuff with a single goal: securing their future. Their message was clear: back-to-basics works, and now it's time to grow the business. Executives from 23 providers outlined their work on increasing revenue, boosting margins and shoring up balance sheets. They presented their market positions, strategic initiatives and financial results for investment analysts, credit raters and bond traders. The third annual Non-Profit Healthcare Investor Conference was co-sponsored by the American Hospital Association, Health Forum, the Healthcare Financial Management Association and Salomon Smith Barney. The focus on growth doesn't come without challenges Several common themes emerged from the presentations, among them pressure on reimbursement, workforce shortages, liability insurance issues and capacity constraints. Yet executives agree: it all comes down to the basics, and building the strength to keep their missions alive. As Thomas Meier, vice president and treasurer of Oakland, Calif.-based Kaiser Permanente, put it: "No margin, no mission, no mas." PMID- 12080923 TI - Info tech's next step. What's driving the race to market this new generation of clinical systems? AB - A new generation of clinical IT systems has hit the market all at once. Is this vigorous competition vendor driven, or has health care--and the technology needed to support it--fundamentally changed? And what does it all mean for hospitals struggling to decide where to spend the big bucks it takes to remain technologically viable? PMID- 12080924 TI - Closing the gap. A new consortium seeks to end health care disparities. AB - New evidence indicates that minorities receive lower-quality health care than whites do, even accounting for economic differences. A new coalition intends to do something about it. PMID- 12080925 TI - Early development of the adrenal glands in the grass snake Natrix natrix L. (Lepidosauria, Serpentes). AB - The aim of the study was to investigate the development and differentiation of the adrenal glands in the grass snake (Natrix natrix L.) during the early stages of ontogenesis, i.e., from egg-laying to hatching of the first specimens. The material used for the studies consisted of a collection of embryos of the grass snake. The Natrix eggs were incubated in the laboratory at a constant temperature of 30 degrees C and 100% relative humidity. Embryos were isolated in a regular sequence of time from egg-laying to hatching. The age of the embryos was qualified according to normal tables for this species. For histological and histochemical investigations, the smallest embryos were fixed in toto. From the oldest embryos, the medial region with the mesonephros and adrenal primordium were resected. Depending on the requirements of histochemical methods, the material was fixed in various fixatives, namely, 10% formalin solution, Bouin, Wood and Millonig fluid, embedded in paraffin and sectioned into serial transversal, sagittal and longitudinal sections. The sections for review were stained with H&E and azan. For detection of adrenaline and noradrenaline in chromaffin tissue, the Wood and Honore methods were used. SGC cells were detected with the silver stain method after Bodian. For electron microscopic studies, the adrenal gland was fixed in 2.5% glutaraldehyde and 2.0% paraformaldehyde 1:1 in 0.1 M phosphate buffer at pH 7.4 and post-fixed in 1.5% osmic acid in the same buffer. The fixed sections of the adrenal glands were embedded in Epon 812. Semithin and ultrathin sections were cut on ultramicrotome ultratome IV. Semithin sections were stained with methylene blue and ultrathin sections were routinely contrasted with uranyl acetate and lead citrate, then examined and photographed with the JEM JEOL 1220 electron microscope. According to morphological and metrical observation in the course of the grass snake embryo development, one can distinguish 12 stages of development. The primordia of the adrenal cortex appear at the first trimester of egg incubation as two asymmetrical strands between the mesonephros and aorta dorsalis. They are made of dense mesenchymal cells. At the second trimester of development, primordia are penetrated by chromaffinoblasts and capillaries. The mesenchymal cells differentiate into interrenal cells, while chromaffinoblasts are arranged dorsally of the gland. The glands are enclosed by the capsule which separates them from the mesonephros. At the third trimester of the eggs incubation, only noradrenaline appears in a chromaffin tissue. At the moment of snake hatching, the adrenal glands are completely differentiated, both in their structure and their function. The primordia of the interrenal tissue differentiate from mesenchymal cells similarly to mammals. During the development of the snake interrenal tissue, several types of cells can be recognized, varying in the degree of differentiation and in ultrastructural features: 1. Undifferentiated cells with features of mesenchymal cells 2. Differentiating mesenchymal cells 3. Transitional cells with features of mesenchymal and steroidogenic cells 4. Differentiating interrenal cells with pleomorphic mitochondria and numerous lipid droplets 5. Embryonic interrenal cells containing circular lipid droplets and underdeveloped smooth endoplasmic reticulum 6. Transitional interrenal cells containing mitochondria with tubular and vesicular cristae, large lipid droplets, numerous myelin structures, and well-developed smooth endoplasmic reticulum 7. Degenerating cells of embryonic interrenal tissue 8. Differentiating mesenchymal cells with features of fibroblasts The above classification is very schematic and presumptive. In developing adrenal glands at the first trimester of egg incubation type 1 and 2 cells predominate. Type 3 and 4 cells were observed at the second trimester of the adrenal primordia development. At the third trimester of egg incubation, embryonic adrenal glands were composed of the type 5 cells. At the moment of snake hatching, interrenal tissue contained type 5 and 6 cells. In the next days of the adrenal gland development, at the border between the cortex and in medulla as under the capsule, numerous cells were degenerated. During the entire development period the adrenal capsule was built from type 7 cells. The chromaffin tissue of the adrenal glands is derived from the neural crest. These findings agree with the findings of all embryologists. The first chromaffinoblasts infiltrated the adrenal cortex primordium around stage IV of development. They were mixed with interrenal cells and just at hatching they were localized dorsally of the gland. The chromaffinoblasts differentiated gradually from neuron-like cells to typical chromaffinocytes. All the chromaffinoblasts contained the chromaffin granules. The size and numerical density of the chromaffin granules increased with development. Just before hatching, the cells of the chromaffin tissue contained only noradrenaline. Differentiation chromaffinoblasts into chromaffin cells are probably stimulated and controlled by the influence of hormones, which are produced by the cells of the interrenal tissue. According to morphological, histochemical and ultrastructural observation in the course of adrenal differentiation and development in the grass snake, six morphological phases can be distinguished. PMID- 12080926 TI - What to look for in HIPAA technology upgrades. PMID- 12080927 TI - First-time CRA exam to be offered at annual meeting. PMID- 12080928 TI - Employee retention: a customer service approach. AB - Employee retention is a huge problem. There are staff shortages in radiology because not enough people are entering the profession; too many people are leaving the profession for retirement, higher-paying jobs or jobs with less stress; and there are not enough opportunities for career advancement. Staff shortages are exacerbated by difficulty in retaining people who enter the profession. While much work has been focused on recruitment and getting more people "in the front door," I suggest that the bulk of future efforts be focused on employee retention and "closing the back door." Employee retention must be an ongoing process, not a program. Approaches to employee retention that focus on external things, i.e., things that the company can do to or for the employee, generally are not successful. The truth is that employee retention processes must focus on what the employee gets out of the job. The process must be a benefits based approach that helps employees answer the question, "What's in it for me?" The retention processes must be ongoing and integrated into the daily culture of the company. The best way to keep your employees is to treat them like customers. Customer service works for external customers. We treat them nicely. We work to satisfy them. We help them achieve their goals. Why not do the same for our employees? If positive customer service policies and practices can satisfy and keep external customers, why not adapt these policies and practices for employees? And, there is a service/satisfaction link between employee retention and higher levels of customer satisfaction. Customers prefer dealing with the same employees over and over again. Employee turnover destroys a customer's confidence in the company. Just like a customer does not want to have to "train and educate" a new provider, they do not want to do the same for your "revolving door" employees. So, the key is to keep employees so they in turn will help you keep your customers. Because the techniques of this process mirror the activities of customer service and customer relationship management, I call the combined process C/ERM for customer/employee relationship management. Both activities must be going on simultaneously to create a loyalty link that ensures customer satisfaction and retention through employee service, satisfaction and retention. PMID- 12080929 TI - The case for RIS/PACS integration. AB - Why integrate PACS with the RIS? To improve workflow, of course, but what workflow? Much of the focus is on improving the flow of images for the radiologist, which is certainly a good thing to do, but what about the rest of the order process? Typical PACS system architecture begins with the HIS since this is where the correct patient demographic information and in many cases the orders originate. Correct patient and order information is sent from the HIS to the RIS using HL7 commands for Admission/Discharge/Transfer (ADT) and Order/Entry. HL7 is the communications protocol used in virtually all information systems. For the first step in communicating with PACS, patient and order information from the RIS is sent to a device called a PACS broker. This is necessary because most PACS systems do not support HL7 directly, and a translation is required. Images from each imaging modality are also sent to the broker using the DICOM standard. If an imaging modality does not support DICOM, then an additional box is used to convert the images to a DICOM file. The broker then sends completed DICOM files to the PACS for storage, distribution and viewing. That approach has worked well for the first stage of PACS utilization. However, experienced PACS users have identified the need to improve workflow, and many feel that closer communication with the RIS will solve many of the current limitations. This approach is sometimes called a "brokerless" solution but is probably better described as incorporating broker functions into the RIS. There are several potential advantages of incorporating the broker functions into the RIS: Access to all RIS information on patients, orders and results is available and can be used in many ways to improve workflow. Supporting all DICOM services directly from the RIS ensures that the latest and most complete information is always used. For example, DICOM Modality Worklists can be provided directly from the RIS, which guarantees that they are updated immediately. The RIS can manage the complete order workflow, not just images. License, implementation and support costs can be reduced by eliminating HL7 interfaces to an external broker. Managing workflow is the key to improved productivity and patient care from PACS. However, coordinated management of order workflow from the RIS and image workflow from the PACS is required to get the full benefit. The RIS has immediate and broad access to patient and order information. As a result, it is the natural place to take the lead in managing this coordinated workflow. While many older RIS and PACS systems are not yet capable of some of the integration features described above, several new systems are moving rapidly in that direction. PMID- 12080930 TI - Digital image archiving: challenges and choices. AB - In the last five years, imaging exam volume has grown rapidly. In addition to increased image acquisition, there is more patient information per study. RIS PACS integration and information-rich DICOM headers now provide us with more patient information relative to each study. The volume of archived digital images is increasing and will continue to rise at a steeper incline than film-based storage of the past. Many filmless facilities have been caught off guard by this increase, which has been stimulated by many factors. The most significant factor is investment in new digital and DICOM-compliant modalities. A huge volume driver is the increase in images per study from multi-slice technology. Storage requirements also are affected by disaster recovery initiatives and state retention mandates. This burgeoning rate of imaging data volume presents many challenges: cost of ownership, data accessibility, storage media obsolescence, database considerations, physical limitations, reliability and redundancy. There are two basic approaches to archiving--single tier and multi-tier. Each has benefits. With a single-tier approach, all the data is stored on a single media that can be accessed very quickly. A redundant copy of the data is then stored onto another less expensive media. This is usually a removable media. In this approach, the on-line storage is increased incrementally as volume grows. In a multi-tier approach, storage levels are set up based on access speed and cost. In other words, all images are stored at the deepest archiving level, which is also the least expensive. Images are stored on or moved back to the intermediate and on-line levels if they will need to be accessed more quickly. It can be difficult to decide what the best approach is for your organization. The options include RAIDs (redundant array of independent disks), direct attached RAID storage (DAS), network storage using RAIDs (NAS and SAN), removable media such as different types of tape, compact disks (CDs and DVDs) and magneto-optical disks (MODs). As you evaluate the various options for storage, it is important to consider both performance and cost. For most imaging enterprises, a single-tier archiving approach is the best solution. With the cost of hard drives declining, NAS is a very feasible solution today. It is highly reliable, offers immediate access to all exams, and easily scales as imaging volume grows. Best of all, media obsolescence challenges need not be of concern. For back-up storage, removable media can be implemented, with a smaller investment needed as it will only be used for a redundant copy of the data. There is no need to keep it online and available. If further system redundancy is desired, multiple servers should be considered. The multi-tier approach still has its merits for smaller enterprises, but with a detailed long-term cost of ownership analysis, NAS will probably still come out on top as the solution of choice for many imaging facilities. PMID- 12080931 TI - Planning for the Mercy Center for Breast Health. AB - During the last months of 2000, administrators at the Mercy San Juan Medical Center in Carmichael, Calif., convened a steering committee to plan the Mercy Center for Breast Health. The Steering Committee was composed of the director of ancillary and support services, the oncology clinical nurse specialist, the RN manager of the oncology nursing unit, the RN surgery center manager, and me, the manager of imaging services. The committee was responsible for creating a new business with five specific objectives: to position the Center as a comprehensive diagnostic and resource center for women; to generate physician referrals to the Breast Center through various vehicles; to create awareness of the Breast Center's capabilities among area radiologists; to create awareness of the Breast Center among employees of six sister facilities; to create "brand awareness" for the Mercy Center for Breast Health among referring physicians and patients who could use competing centers in the area. The Steering Committee's charter was to design a center with a feminine touch and ambience and to provide a "one-stop shopping" experience for patients. A major component of the Breast Center is the Dianne Haselwood Resource Center, which provides patients with educational support and information. The Steering Committee brought its diverse experience and interests to bear on arranging for equipment acquisition, information and clerical systems, staffing, clinic office design, patient care and marketing. Planning the Mercy Center for Breast Health has been a positive challenge that brought together many elements of the organization and people from different departments and specialties to create a new business venture. Our charge now is to grow and to live up to our vision of offering complete breast diagnostic, education and support services in one location. PMID- 12080932 TI - The financial importance of medical imaging. PMID- 12080933 TI - And the sign said, "Y chromosomes need not apply". PMID- 12080934 TI - The more things change,.... PMID- 12080935 TI - [Corticosteroids for treating Kawasaki disease]. PMID- 12080936 TI - Patient Group Directions for midwives. PMID- 12080937 TI - Ready, steady, change! PMID- 12080938 TI - Chemokine/chemokine receptor nomenclature. PMID- 12080939 TI - Open your mouth. PMID- 12080940 TI - Photo quiz. Huntley's papules (diabetic finger pebbling). PMID- 12080941 TI - Congenital triangular alopecia: a case report and review. AB - Congenital triangular alopecia is a nonscarring loss of hair mass on the scalp's temporal regions. The area of hair diminution commonly is described as triangular or lancet shaped. Although previously considered congenital, this condition usually is noticed after 2 years of age and, more recently, is thought to be acquired. We propose that this entity be renamed triangular alopecia. Because this condition involves normal rather than inflamed skin, it does not respond to topical or intralesional steroids. It is important to make the correct diagnosis to avoid unnecessary and potentially harmful interventions. We present the case of a 10-year-old boy with triangular alopecia. PMID- 12080942 TI - What's eating you? Latrodectus mactans (the black widow spider). PMID- 12080943 TI - Pyoderma faciale: gram-negative recovery by means of needle aspiration. AB - Pyoderma faciale is an inflammatory disease that until now was believed to occur only in women. Bacterial pathogens have not been found in most cases, and those found included only gram-positive microorganisms. We present 2 cases consistent with pyoderma faciale--one of them a 17-year-old adolescent boy--in which gram negative bacteria were found. Enterobacter cloacae was found in one patient, and Klebsiella oxytoca was found in the other. Needle aspiration is the technique of choice in looking for pathogens in this disease. PMID- 12080944 TI - Bullae and sweat gland necrosis after an alcoholic deep slumber. AB - A 37-year-old man developed edematous areas and blisters on the right side of his face, chest, and arm after an alcoholic deep slumber. It was revealed that the affected body parts were those pressed during his alcoholic sleep. Histopathological findings of the patient's skin lesions showed typical sweat gland necrosis. Serum enzyme level studies of aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and creatine phosphokinase were characteristic of muscular damage. This case report is an example of the typical findings of the effects of body pressure on soft tissue that can be seen in a dermatology clinic. PMID- 12080945 TI - Henoch-Schonlein purpura secondary to subacute bacterial endocarditis. AB - Henoch-Schonlein purpura (HSP), a systemic, small-vessel vasculitic syndrome, is characterized by a nonthrombocytopenic purpuric rash, arthralgia, abdominal pain, and nephritis. These signs and symptoms may occur in any order, and not all are necessary for the diagnosis. Although most common in 4- to 7-year-olds, HSP is well documented in adults and is often preceded by a history of mucosal-based infections, especially of the upper respiratory tract. We report a case of HSP that occurred coincident with the onset of subacute bacterial endocarditis (SBE) in an otherwise healtny 41-year-old white woman. The patient presented with a purpuric rash and arthralgia and was found to have left-sided streptococcal SBE. She subsequently developed abdominal pain and immune complex glomerulonephritis. The bacterial endocarditis was treated with antibiotics and mitral valve replacement, followed by a spontaneous resolution of the associated signs and symptoms of HSP. PMID- 12080946 TI - An epidemicity of Paederus species in Cukurova region. AB - Two hundred four patients (117 females, 87 males; age range: 3-80 y) were admitted to our facility between May 1995 and June 1997 and studied to determine the endemicity of the Paederus species, which has been increasing for the last 6 years (especially in May and June) in the Cukurova region of southern Turkey. Clinically, infection with the Paederus species mimics contact dermatitis, herpes zoster, bullous impetigo, and phytophotodermatitis. Definitive diagnosis is made by historical and clinical findings. To determine the main histopathologic features of this infestation, biopsy specimens were obtained from 9 patients and stained with hematoxylin and eosin (H&E). In most patients, the skin lesions were located on the exposed parts of the body. Clinically, these lesions were linear, vesicular, bullous, and/or pustular on erythematous bases and resembled either phytophotodermatitis, herpes zoster, or impetigo rather than classic insect bites. Pederin, which is released from the Paederus species, may cause these lesions. The number of cases has increased markedly during the last 5 years. In the coming years, we expect this number to increase significantly. PMID- 12080947 TI - Centrifugal lipodystrophy presenting with serpiginous erythema and alopecia. AB - We describe serpiginous erythema with alopecia developing on the scalp of a 10 year-old boy during follow-up of centrifugal lipodystrophy. Because the clinical and histopathologic features of these lesions were identical to those of centrifugal lipodystrophy, we conclude that involvement of a hairy region by this disorder could cause alopecia and that the hair loss might be an indirect effect of interstitial inflammatory infiltrates around the hair follicles and in the subcutaneous fat. PMID- 12080948 TI - Lichen sclerosus: an atypical presentation. AB - We describe the case of a 66-year-old Hispanic man with an atypical presentation of lichen sclerosus (LS). The unusual presentation included bilateral axilla involvement (not previously reported to our knowledge), scrotal involvement (not common in men, despite common vulvar involvement in women), and an uncommonly thick plaque on his back. PMID- 12080949 TI - Nodulocystic acne and hidradenitis suppurativa treated with acitretin: a case report. AB - Nodulocystic acne is a dermatologic disease that can result in significant damage to the skin of the face, chest, and back. Hidradenitis suppurativa is a scarring disease of the skin that causes deep cysts and abscesses on the axillae and anogenital areas. We review a case of a patient with severe nodulocystic facial acne and hidradenitis suppurativa that was treated with 2 full courses of isotretinoin. Although the patient's condition improved, some draining cysts persisted on the face and groin. Because of the inability of isotretinoin to achieve long-term remission of the patient's condition, acitretin was considered as a possible maintenance drug. The patient was almost completely improved after 5 months' therapy with acitretin, which also was effectively used for ongoing maintenance. Acitretin may be a promising treatment for severe nodulocystic acne and hidradenitis suppurativa, which require long-term suppression when isotretinoin fails to give long-term remission. PMID- 12080950 TI - Herpetic finger infection. AB - We present a case of herpetic gingivostomatitis and finger infection. Vesicular hand lesions may result from autoinoculation of oral herpes simplex virus (HSV) infection in children, which may be evident or asymptomatic. PMID- 12080951 TI - Aleukemic monocytic leukemia cutis. AB - Aleukemic leukemia cutis is a rare condition in which leukemic cells invade the skin before they appear in peripheral blood or bone marrow specimens. The condition frequently is misdiagnosed as atypical lymphoma. Generally, the diagnosis is made retrospectively, after the leukemic cells appear in peripheral blood or bone marrow samples. Immunohistochemical studies are the primary methods for diagnosis. Prognosis is usually poor. We describe the case of a 75-year-old woman with acute aleukemic monocytic leukemia cutis who developed systemic disease 1 1/2 years after skin involvement. PMID- 12080952 TI - Superficial mycosis superimposing on isolated lichen planus of the lip: a case report and review of the literature. AB - We report a case of superficial mycosis superimposing on isolated lichen planus (LP) of the lower lip that was successfully treated. A 36-year-old woman had 2 ulcerated, scaling, erythematous plaques on the lower lip for several months with mild painful sensation. Results of histologic study showed typical features of LP, but fungal hyphae were found in the horny layer. The lesion healed with only very mild residual erythema after 6 weeks of treatment. A review of the literature shows only one noted case of tinea of the lip, which was reported in 1968 by a dental practitioner, and only several cases of isolated LP of the lip. The possible mechanism of coexistence of tinea and LP of the lip is reviewed. PMID- 12080953 TI - Linear porokeratosis with underlying bony abnormalities. AB - A 58-year-old woman with linear porokeratosis involving the right hand and arm had distal digital narrowing and nail dystrophy with radiographic changes. Whereas isolated cases of bone resorption and flexion deformities with porokeratosis of Mibelli are known to occur, to our knowledge, bony abnormalities in association with linear porokeratosis have not been reported. PMID- 12080954 TI - Body odor in dermatologic diagnosis. PMID- 12080956 TI - Proper HIV/AIDS care not possible without basic safety in health set-up. PMID- 12080955 TI - Preventing cardiovascular disease. A despondent view. PMID- 12080958 TI - Withdrawal of "Caveolae and their coat proteins, the caveolins: from electron microscopic novelty to biological launching pad". PMID- 12080957 TI - Fabrazyme--recombinant protein treatment for Fabry's disease. PMID- 12080959 TI - In vitro demonstration of anaerobic oxidation of methane coupled to sulphate reduction in sediment from a marine gas hydrate area. AB - Anaerobic oxidation of methane (AOM) and sulphate reduction were examined in sediment samples from a marine gas hydrate area (Hydrate Ridge, NE Pacific). The sediment contained high numbers of microbial consortia consisting of organisms that affiliate with methanogenic archaea and with sulphate-reducing bacteria. Sediment samples incubated under strictly anoxic conditions in defined mineral medium (salinity as in seawater) produced sulphide from sulphate if methane was added as the sole organic substrate. No sulphide production occurred in control experiments without methane. Methane-dependent sulphide production was fastest between 4 degree C and 16 degree C, the average rate with 0.1 MPa (approximately 1 atm) methane being 2.5 micro mol sulphide day(-1) and (g dry mass sediment)( 1). An increase of the methane pressure to 1.1 MPa (approximately 11 atm) resulted in a four to fivefold increase of the sulphide production rate. Quantitative measurements using a special anoxic incubation device without gas phase revealed continuous consumption of dissolved methane (from initially 3.2 to 0.7 mM) with simultaneous production of sulphide at a molar ratio of nearly 1:1. To test the response of the indigenous community to possible intermediates of AOM, molecular hydrogen, formate, acetate or methanol were added in the absence of methane; however, sulphide production from sulphate with any of these compounds was much slower than with methane. In the presence of methane, such additions neither stimulated nor inhibited sulphate reduction. Hence, the experiments did not provide evidence for one of these compounds acting as a free extracellular intermediate (intercellular shuttle) during AOM by the presently investigated consortia. PMID- 12080960 TI - [Ivory watchers]. PMID- 12080961 TI - Updated results of a randomized, double-blind, multicenter sham-controlled trial of microwave thermotherapy with the Dornier Urowave in patients with symptomatic benign prostatic hyperplasia. Urowave Investigators Group. AB - A total of 220 patients with clinical benign prostatic hyperplasia (BPH), an AUA symptom index (AUA SI) of > 13 points, and a peak flow rate of < 12 ml/s were randomly assigned to either active or sham treatment. All treatments were conducted as an outpatient procedure without general anesthesia. Sham-treated patients underwent a simulated 60-min treatment with an indwelling treatment catheter. Patients were followed at 1 week and at 1, 3, and 6 months. The treatments were well tolerated. The AUA SI dropped from 23.6 to 12.6 points at 6 months (P < 0.05) in the active group and from 23.9 to 17.9 points in the sham treated group (P < 0.05 for the difference between groups). There was a significant difference between the groups at 6 months (P < 0.001). Similar and statistically significant changes occurred regarding the AUA bother index and the quality of life scores. Statistically significant improvements were also noted for the peak flow rate (from 7.7 to 10.6 ml/s at 6 months for the active group and from 8.1 to 9.6 ml/s for the sham-treated group; P < 0.05 for the difference between groups) and for the average flow rate. An improvement in the AUA SI of > 30% was achieved by 72% versus 40% (active versus sham) and an improvement of > 50% was accomplished by 48% versus 21% of the respective patients. Actively treated patients reported more dysuria, urgency, and ejaculatory dysfunction following treatment than did sham-treated patients. Urinary retention occurred in 5.4% of patients. The Dornier Urowave is better in improving symptoms, bother, quality of life, and flow rates than is a sham treatment in patients with symptomatic BPH. The treatments can be given on an outpatient basis with local anesthesia. Adverse events are in general transient and mild in nature. This device compares favorably with other second-generation devices. Extended follow up is necessary to document the long-term durability of these improvements. PMID- 12080962 TI - Noncontact detection of ultrasonic waves using fiber optic Sagnac interferometer. AB - This paper describes a fiber optic sensor suitable for noncontact detection of ultrasonic waves. This sensor is based on the fiber optic Sagnac interferometer, which has a path-matched configuration and does not require active stabilization. Quadrature phase bias between two interfering laser beams in the Sagnac loop is applied by controlling the birefringence using a fiber polarization controller. A stable quadrature phase bias can be confirmed by observing the interferometer output according to the change of phase bias. Additional signal processing is not needed for the detection of ultrasonic waves using the Sagnac interferometer. Ultrasonic oscillations produced by conventional ultrasonic piezoelectric transducers were successfully detected, and the performance of this interferometer was investigated by a power spectrum analysis of the output signal. Based on the validation of the fiber optic Sagnac interferometer, noncontact detection of laser-generated surface waves was performed. The configured Sagnac interferometer is very effective for the detection of small displacement with high frequency, such as ultrasonic waves used in conventional nondestructive testing (NDT). PMID- 12080963 TI - [Adaptation of Child Abuse Potential Inventory in Chile: analysis of the difficulties and challenges in the application in Chilean studies]. AB - OBJECTIVE: This paper describes two studies aimed at validating the Chilean version of Child Abuse Potential (CAP). The paper also discusses problems in what respects conceptualizing and detecting maltreatment within a process of research as it relates to a specific culture. METHOD: An instrument based on the Child Abuse Potential Inventory was administered to two different samples of 134 people and 280 people, respectively. Both studies are contrasted in terms of internal consistency, factor analysis, and the instrument's ability to discriminate between abusers and non-abusers. RESULTS: Both studies feature good internal consistency (.98 and .95) and a factor structure similar to that of the original instrument; however, the second study shows problems concerning correct subject classification. The contrast of both studies suggests that the instrument is suitable for discriminating between severe child physical abuse and very well treated children. The indecision area of classification is 67%. CONCLUSION: The results of both studies highlight the need to conceptualize and operationalize child abuse within a specific cultural context. The distinction between abusers and non-abusers becomes more blurred in the less extreme cases. PMID- 12080965 TI - Africa can solve its own health problems. . . but can it solve these problems itself? PMID- 12080964 TI - Renal Transplantation. Cadaveric heart beating donor rates vary from region to region in United Kingdom. PMID- 12080966 TI - Denmark: debate outline on fetal diagnostics (1999). PMID- 12080967 TI - The European protocol on organ transplantation does not meet public expectations. PMID- 12080968 TI - Justice and equity in transplantation medicine. PMID- 12080969 TI - Draft protocol on transplantation of organs and tissues of human origin. PMID- 12080970 TI - Draft explanatory report to the draft protocol. PMID- 12080971 TI - Diagnosis of malaria using thick bloodsmears: definition and evaluation of a faster protocol with improved readability. AB - The value of some inexpensive modifications to the standard method of preparing thick bloodsmears, involving rapid drying, an isotonic fixative and a haemolysing solution containing saponin, was evaluated. The drying, haemolysing, fixing and staining steps, together called the fast-thick-smear method (FTS), can be completed in < 10 min. The FTS and a more classical thick-smear method (CTS) were both used on each of 1185 samples of venous blood samples from 1034 cases of suspected malaria (all international travellers returning to France). The results indicated that there was no statistically significant differences between the two methods in terms of their sensitivity, specificity or predictive values for parasite detection. However, estimates of the intensities of the Plasmodium falciparum infections observed, based on counts of trophozoites against 200 leucocytes, were markedly higher (37.8% higher overall) with the FTS than with the CTS (P < 0.0001). Moreover, the concordance between results obtained by inexperienced and experienced microscopists was excellent when the FTS was used, with a kappa value of 0.96 (95% confidence interval = 0.93-0.98). PMID- 12080972 TI - Cardiac denervation in mice infected with Trypanosoma cruzi. AB - The neuronal features of the hearts of mice that were acutely or chronically infected with the Y strain of Trypanosoma cruzi were compared with those of control hearts from uninfected mice. Whole-mount preparations of the murine atria, isolated by microdissection, were stained to reveal neurons with NADH diaphorase activity. Counts, by a microscopist who was blind to the infection status of the donor mouse, revealed that there were significantly (38%) fewer such neurons in the atria from the acutely infected mice than in the atria from the control hearts. The ganglia of the infected mice were also irregularly distributed, severely damaged ganglia being found beside slightly degenerated or morphologically normal ones. Although the ganglia contained small, medium and large neurons, the apparent destruction caused by T. cruzi was confined to the large ones. As neuron counts in preparations of hearts from mice with chronic infections were 32% lower than those in the control hearts, there appears to be no additional loss of cardiac neurons as the acute infection in mice progresses to the chronic phase. PMID- 12080973 TI - The rarity of infection with Leishmania (Viannia) braziliensis among patients from the Manaus region of Amazonas state, Brazil, who have cutaneous leishmaniasis. AB - The frequency of Leishmania ( Viannia) braziliensis infection was assessed in 79 of the 138 patients with cutaneous leishmaniasis who attended a reference outpatient unit in Manaus, Amazonas state, between the August and December of 1997. The disease was characterized by one or more cutaneous ulcers, the skin lesions being frequently associated with satellite lymph-node enlargement. All parasite isolates were identified using monoclonal antibodies and enzyme electrophoresis. Only two (2.8%) of the 71 patients from whom parasites were successfully isolated were found to be infected with L. ( V.) braziliensis, the other 69 isolates being identified, from their isoenzyme profiles, as L. ( V.) guyanensis. In the Manaus region, therefore, almost all human cutaneous leishmaniasis is the result of infection with L. (V.) guyanensis, and L. ( V.) braziliensis is a relatively rare cause of the disease. PMID- 12080975 TI - Comparative cost-effectiveness of diagnostic tests for urinary schistosomiasis and the implications for school health programmes. AB - The use of self-reported schistosomiasis or blood in urine has received a great deal of interest as a cheap and simple technique for diagnosing individuals infected with Schistosoma haematobium and identifying schools with a high prevalence of infection. Although the answers to questions about the signs and symptoms of urinary schistosomiasis have been shown to be good markers of parasitological infection, a formal cost-effectiveness analysis of their performance in relation to urine filtration and parasitological examination (assumed to be the gold standard) is lacking. Using empirical data on the costs and effectiveness of these techniques in 15 schools in Tanzania, the cost for every correct diagnosis or for every infected child identified was assessed. Although self-reported schistosomiasis was shown to be three times more cost effective than urine filtration in identifying infected individuals, it would have resulted in a third of the infected children being missed. Use of self reported schistosomiasis first to identify high-risk schools for mass treatment and then to identify infected children in low-risk schools (for individual treatment) also appeared more cost-effective than urine filtration and would have resulted in only 8% of the infected children not being treated. The use of self reported schistosomiasis or self-reported blood in urine should be continually supported as a simple, cheap and cost-effective tool for identifying schools at high-risk of schistosomiasis. PMID- 12080974 TI - Lymphatic filariasis in the Karonga district of northern Malawi: a prevalence survey. AB - In Malawi, two main foci of lymphatic filariasis (LF) are known to exist: one in the south, in the Shire valley, and the other in the north, along the Songwe River, on the border with Tanzania. There have been no formal surveys in the Songwe area since the 1960s but an opportunity arose in 2000-2001 to map LF in this area, in the context of a leprosy survey that formed part of the follow-up of a large leprosy and tuberculosis vaccine trial. Overall 687 immunochromatographic (ICT) tests were carried out. Wuchereria bancrofti antigenaemia was found in > 25% of adults in each of the 12 villages sampled (four in the Songwe area and eight in the rest of the Karonga district), with village prevalences varying from 28%-58%. Of the 685 adult male residents of the Songwe area who were each given full-body clinical examinations, 80 (11.7%) were identified as cases of hydrocele. Lymphoedema was found in seven (1.0%) of these adult males and in 29 (3.7%) of the 769 adult female residents of the Songwe area who were also examined. Microfilariae were detected in 33 (30.8%) of the 107 thick smears of night-blood samples that were made from individuals with positive ICT cards. The W. bancrofti infection focus in Karonga district is therefore wider than was previously known. This has important implications for the implementation and eventual impact of LF-control activities in this area. PMID- 12080976 TI - Revisiting the question of limited genetic variation within Schistosoma japonicum. AB - Recent electrophoretic data have indicated that Schistosoma japonicum in mainland China may be a species complex, with the existence of a cryptic species being predicted from the analysis of schistosome populations from Sichuan province. To investigate the Sichuan form of S. japonicum, 4.9 kbp of mitochondrial DNA from each of three samples of the parasite from China (two from Sichuan and one from Hunan) and one from Sorsogon in the Philippines were amplified, sequenced and characterized. The sequence data were compared with those from the related South east Asian species of S. mekongi (Khong Island, Laos) and S. mlayensis (Baling, Malaysia) and that from S. japonicm from Anhui (China). At both the nucleotide and amino-acid levels, the variation among the five S. japonicum samples was limited (< 1%). This was consistent with the conclusions drawn from previous molecular studies, in which minimal variation among S. japonicum populations was also detected. In contrast, S. mekongi and S. malayensis, species recognized as separate but closely related, differ from each other by about 10%, and each differs by 25%-26% from S. japonicum. Phylogenetic trees provided a graphic representation of these differences, showing all S. japonicum sequences to be very tightly clustered and distant from S. mekongi and S. malayensis, the last two being clearly distinct from each other. The results thus indicate no significant intra-specific genetic variation among S. japonicum samples collected from different geographical areas and do not support the idea of a distinct form in Sichuan. PMID- 12080977 TI - Snail-parasite compatibility and prevalence of Schistosoma haematobium on the shores of Lake Kariba, Zambia. AB - Reduced snail-parasite compatibility might be at least partially responsible for the decrease observed, over the last three decades, in the prevalence of urinary schistosomiasis on the shores of Lake Kariba, Zambia. To explore this possibility, the prevalences of urinary schistosomiasis in schoolchildren were investigated in three disparate areas of Zambia (Lake Kariba, Lake Bangweulu and Lusaka), and attempts were made to infect the snails that developed from the eggs of snails collected from each of these sites with Schistosoma haematobium from each of the sites. The prevalence of S. haematobium infection in schoolchildren ranged from 0% around Lake Bangweulu to 76% around Lake Kariba. The F1 progeny of Siavonga (Lake Kariba) snails showed good compatibility with the local parasite but were also susceptible to all of the geographical strains of S. haematobium tested. The interaction between the S. haematobium and S. mansoni found along the shores of Lake Kariba may favour S. mansoni, since prevalences of urinary schistosomiasis tend to be relatively high where S. mansoni is absent. Although Lake Bangweulu was confirmed to be an area of very low endemicity for urinary schistosomiasis, the snails bred from snails collected in this area were compatible with isolates of S. haematobium from the same region and also those from the other regions. In general, sympatric snail-parasite combinations were the most successful. All the snails were identified as Bulinus globosus, on the basis of common enzyme profiles, although polymorphism was evident for malate dehydrogenase (MDH): two samples had type-1 MDH and the rest were of type 3. Isoenzyme patterns for both acid phosphatase (AcP) and MDH could be used to distinguish between infected and uninfected snails. PMID- 12080978 TI - Implication of Bulinus truncatus in the transmission of urinary schistosomiasis in Senegal, West Africa. AB - Studies on the compatibility between Bulinus truncatus and Schistosoma haematobium from various areas in the Senegal River basin (SRB) were carried out, to investigate the role of B. truncatus in the epidemiology of urinary schistosomiasis in Senegal. The results show that B. truncatus from the Lower Valley is not compatible with S. haematobium from the Middle Valley. Interestingly, the study reveals that B. truncatus from the Upper Valley is compatible with S. haematobium from the same area, and that S. haematobium from the Middle Valley is compatible with B. truncatus from the Lower Valley. Bulinus truncatus naturally infected with S. haematobium have been collected from the Upper Valley of the SRB. These results indicate that B. truncatus is involved in the urinary schistosomiasis in Senegal. PMID- 12080979 TI - Excess mortality associated with blindness in the onchocerciasis focus of the Mbam Valley, Cameroon. AB - The association between blindness, mortality and nutritional status was investigated in a retrospective cohort study in villages of central Cameroon where onchocerciasis is hyper-endemic. Overall, 101 blind subjects and 101 non blind controls matched with the blind for age, sex and (generally) village of residence were followed for 10 years. Blindness gave rise to a significant increase in mortality (relative risk = 2.3; P = 0.012), the life expectancy of the blind adults being reduced by 4 years compared with that of their controls. For a given age, excess mortality was found to be associated with a late onset of blindness. The causes of death were similar for the blind and the controls but blind subjects had relatively low body mass indices, which may lead to relatively early fatal disease outcomes. These results are similar to those obtained in other parts of Africa and emphasise, once more, the demographic impact of blindness in developing countries. PMID- 12080980 TI - A survey of the infective larvae of Gnathostoma spinigerum in swamp eels bought in a local market in Bangkok, Thailand. AB - The livers were separated from the viscera of 2738 swamp eels (Monopterus alba) purchased from Klong Toey market, the largest market in Bangkok, between June 1999 and May 2000. When these livers were digested in artificial gastric juice, 524 (19.1%) were found to be infected with the human-infective, third-stage larvae (L3) of Gnathostoma spp. All the identified larvae were confirmed morphologically to be G. spinigerum. Prevalence of the infection varied with season, from a high of 38.3% in September to a low of 7.0% in April, being generally high during the rainy season and winter (June-February). The mean (S.E.) number of L3 recovered/infected liver, which was 3.99 (0.52) overall, also varied with the season, peaking at 5.38 (1.89) in January, but the month-on-month variation was not statistically significant. Although the results of an earlier study had indicated that the prevalence of eel infection decreased in November, after the rainy season, the most abrupt decrease observed in the present study occurred at the beginning of summer (March). However, the period covered by the present study was unusually wet, and the prevalence of eel infection may depend on rainfall more than season. PMID- 12080982 TI - Are intestinal helminths a risk factor for non-typhoidal Salmonella bacteraemia in adults in Africa who are seropositive for HIV? A case-control study. AB - In Africa, invasive, non-typhoidal Salmonella (NTS) infections are a common but life-threatening complication in adults who are seropositive for HIV. The high prevalence of human infection with intestinal helminths which penetrate the gut could explain the greater importance of NTS bacteraemia in Africa compared with that in industrialized countries. If helminth infection is a major risk factor for NTS it would provide a locally relevant, public-health target. Intestinal helminth carriage in 57 HIV-positive patients with NTS bacteraemia (the cases) was compared with that in 162 HIV-positive controls who were similar to the cases in terms of age, sex, urban dwelling and socio-economic factors. The prevalence of helminth infection, 29% overall, was lower among the cases (18%) than among the controls (33%), giving a crude odds ratio of 0.40 [with a 95% confidence interval (CI) of 0.21-0.9] and an adjusted odds ratio (aOR) of 0.79 (CI = 0.4 1.8). Five (9%) of the cases and 12 (7%) of the controls were infected with nematodes which penetrate the gut (Ascaris lumbricoides and/or Strongyloides stercoralis). The aOR for infection with these penetrating worms, corrected for age, sex, urban dwelling and phase of study, was 1.40 (CI = 0.4-4.5). The present results do not exclude the possibility that helminths play a role in invasive NTS infections, but are not consistent with helminths being a sufficient risk factor in this population to be a public-health target. Anthelmintics are unlikely to have a major impact on preventing NTS bacteraemia in patients diagnosed HIV positive in Africa. PMID- 12080981 TI - Hepatitis-C-virus genotypes and hepatitis-G-virus infection in Lebanese thalassaemics. AB - Exposure to hepatitis C virus (HCV), hepatitis G virus (HGV) and the carrier 'rate' for hepatitis B virus (HBsAg) were investigated in thalassaemia patients in Lebanon, a group that has not been studied in the past. The HCV genotypes and their distribution in the 395 thalassaemics, all of whom had been registered at the Chronic Care Center (CCC) in Hazmieh since 1996, were also studied. Of the 55 samples (14%) found positive for anti-HCV, 19 were also positive for HCV RNA. The 19 samples of HCV RNA were mostly of genotype 4 (37%), followed by 1a and 3a (21% each), lb (16%) and 2b (5%). Most (14; 74%) of the 19 HCV-RNA-positive samples, but only 13 (36%) of the 36 samples that were negative for HCV RNA although anti HCV-positive, were positive for anti-HGV. Among 100 anti-HCV-negative samples, eight (8%) were anti-HGV positive. Only one (0.28%) of all 395 patients investigated was found to be HBsAg-positive. All of the HBV- and HCV-positive patients had initially been found positive in 1996, when they were first registered at the CCC, and none of the remaining patients had seroconverted since. As none of the patients had been checked for anti-HGV until the present study, the history of their exposure to HGV was unknown. These results emphasise the importance of screening all blood donations collected in Lebanon for HBsAg and anti-HCV. This and stringent infection-control measures are necessary steps to limit the spread of HBV, HCV and perhaps HGV to thalassaemics. PMID- 12080983 TI - Molluscicidal activity of plants from Puerto Rico. AB - Overall, 173 tropical plants from 72 different families, collected from the north western and western regions of Puerto Rico, were screened for their molluscicidal properties against Biomphalaria glabrata, the intermediate host of Schistosoma mansoni. Six plant extracts were effective at 50 ppm. The two most effective extracts were those from the leaves of Didymopanax morototoni (Araliaceae) and Mammea americana (Guttiferae), which, at 50 ppm, killed all snails after 24 h of exposure and a day for recovery. Under the same conditions, extracts of Furcraea tuberosa, Argemone mexicana and Paullinia pinnata killed 50% of the snails and that of Solanum americanum killed 33%. The most effective extracts (or their active components or compounds based on them) may have potential as molluscides for the relatively cheap control of human schistosomiasis. PMID- 12080984 TI - Cystic echinococcosis in a rural Peruvian family. PMID- 12080985 TI - Electronic journals: looking ahead. PMID- 12080986 TI - History of the imagery of tympanic membrane pathology: from first drawings to rod lens endoscopic photography. AB - This study was performed to determine the historical development of the techniques used to represent the pathologic conditions of the tympanic membrane. The first coloured attempts were the drawings of Adam Politzer in 1865. Over the following 50 years other otologists produced their own atlases of drawings. At the beginning of the 20th century some otologists tried to photograph the tympanic membrane but without success. This method was progressively improved and by 1960 two techniques were used: through a speculum or with a microscope. The quality was not optimal, and for this reason, atlases produced during this period were with retouched or water-coloured photographs. It was with the discovery of new endoscopic fibres at the beginning of the 1960s that endoscopic photography progressively replaced all other methods. The precision of these pictures is very important for improving the quality of teaching of the different tympanic membrane pathology. PMID- 12080987 TI - Histochemical, immunohistological and scanning electron microscope analysis of tissue retained on spontaneously extruded ventilation tubes. AB - The aim of this study was to analyse the tissue surrounding prematurely extruded ventilation tubes (grommets). Thirty-one ventilation tubes, including 21 Shah and six Shepherd tubes, that had been extruded naturally into the ear canal were examined. After formalin fixation, material adhering to the tubes was retrieved and processed to paraffin wax. Five tubes were processed for scanning electron microscope (SEM) analysis. Sections were stained using histochemical methods for collagen, keratin and keratohyaline. In addition, the presence of collagen (type I and III), keratin, vimentin, fibronectin, tenascin, factor VIII-related antigen, CD31 and CD45 was tested for by immunohistochemistry. Results showed that all specimens consisted of an acellular fibrous material, oriented in concentric rings parallel to the tube surface that was often associated with small collections of CD45+ inflammatory cells. Two specimens contained collagen that was detectable by histochemical and immunohistological methods. Twelve specimens contained identifiable desquamated epithelial cells containing keratin and keratohyaline. Only one specimen stained positively for connective tissue markers (vimentin, fibrous fibronectin, tenascin) or showed the presence of vascular epithelium. SEM revealed adherent clusters or sheets of plate-like structures, consistent with the presence of epithelial squames, on three of the five ventilation tubes examined. It is concluded that tissue retained on extruded grommets consists of orthokeratinitizing epithelium infiltrated by inflammatory cells, a finding that is consistent with the theory that tube extrusion occurs as a result of squamous epithelial proliferation, medial migration, differentiation and desquamation. PMID- 12080988 TI - Staging and management of primary cerebellopontine cholesteatoma. AB - Primary cerebellopontine angle (CPA) cholesteatoma grows slowly and silently in the subarachnoidal spaces. The diagnosis is often late, when the lesion has reached large dimensions. Surgical removal is the only available therapy. Fifteen consecutive cases of CPA cholesteatoma managed at a tertiary otoneurosurgical referral unit between September 1985 and April 1999 were reviewed. The study population, consisting of seven males and eight females, had a mean age of 44 years of age (range 21-69) at the time of surgery. The clinical, audiological and radiological presentations were examined. The tumours were classified according to the Moffat classification of CPA cholesteatomas. In 67 per cent of cases the presenting symptom was related to the vestibulo-cochlear nerve. The average duration of symptoms was 23 months (ranging from one month-10 years). The hearing preservation approaches were utilized the most (11 cases), while the translabyrinthine approach alone, or in association with a middle fossa craniotomy, was performed in four cases. Tumour removal was total in 12 cases and subtotal in three cases. In cases undergoing hearing preservation surgery the mean pre-operative pure tone average (PTA) for the frequencies 0.5, 1, 2, and 3 kHz was 19.3 dB HL (SD 13.84) and the mean pre-operative speech discrimination score (SDS) was 89.8 per cent (SD 5.97). In 44.4 per cent of patients the hearing was preserved and the mean post-operative PTA was 20.29 dB HL (SD 15.84). In five patients post-operative complications occurred. No peri- or post-operative death occurred in this series, one patient developed a recurrence 15 years after the initial surgery. PMID- 12080989 TI - Management of nasal injuries: a postal questionnaire survey of UK ENT consultants. AB - Nasal injuries are one of the commonest conditions seen in ENT out-patients. There is no uniform policy or protocol for the management of this condition. A postal questionnaire was sent to all UK-based ENT consultants. The aim of this study was to ascertain current practice in the management of this common problem. We were also interested to know if consultants believed that the septum should be addressed and dealt with at the same time as manipulation to improve cosmetic and functional results. Manipulation of nasal bones without anaesthesia, under local anaesthesia, and general anaesthesia with digital pressure or using instruments are all practised. The digital pressure under general anaesthesia with use of septal and nasal forceps, if needed, is practised by the majority of consultants (55.8 per cent). Only a minority considered the role of the septum to be important in the initial management of the patient. PMID- 12080990 TI - The prevalence of latex allergy in patients with rhinitis. AB - Natural rubber latex hypersensitivity to the constituent proteins of natural rubber latex is now an international health problem. This study investigates the prevalence of latex allergy in patients presenting with rhinitis. Fifty-nine consecutive patients, who presented to a general Otolaryngology out-patient clinic with symptoms suggestive of rhinitis, completed a questionnaire and underwent skin prick testing. Three out of 59 (5.1 per cent) tested positive to latex. The overall positive rate (all standard aeroallergens) was 45.8 per cent. Our data supports the hypothesis that the prevalence of latex allergy in patients with rhinitis is higher than the general population and that rhinitis may be a presenting symptom. PMID- 12080991 TI - Expression of GSTM1 in angiofibromas. AB - The aetiology of juvenile nasopharyngeal angiofibroma (JNA) is largely unknown. In this study, we have investigated the expression of glutathione S transferase M1 (GSTM1) gene in angiofibroma. The GSTM1 allele gene locus is normally found in all human beings. When this is not expressed there is an increased risk of developing a malignancy of the upper aerodigestive tract. In this study, we have assessed eight samples of JNA for the expression of GSTM1, by the polymerase chain reaction (PCR) followed by polyacrylamide gel electrophoresis (PAGE), three of the eight patients studied failed to express this gene. Further investigation in this area is warranted. PMID- 12080992 TI - Significance of p53 expression in non-tumoral epithelium adjacent to oral squamous cell carcinomas. AB - Alterations of the p53 gene are the most frequently documented genetic abnormalities in human cancer. The aim of the present study was to analyse if this alteration is an early event in oral tumorigenesis and if the suprabasal expression of p53 is a marker of the presence and severity of epithelial dysplasia. Immunohistochemical p53 expression in 78 specimens of oral squamous cell carcinoma and non-tumoral adjacent epithelium was analysed. Non-tumoral epithelium was observed in 53 cases (67.9 per cent), being normal in six cases (7.6 per cent), hyperplastic in 24 cases (30.7 per cent) and dysplastic in 48 cases (61.5 per cent). Epithelial dysplasia was mild (23 cases, 47.9 per cent); moderate (23 cases, 47.9 per cent) and severe (two cases, 4.1 per cent). Twenty one cases of the dysplasias (43.8 per cent) expressed p53. No p53 expression appeared in any normal epithelium. Basal p53 expression always appeared in mild dysplasias (two cases). Suprabasal p53 expression appeared in mild and moderate dysplasias in nine cases and in one severe dysplasia. No statistical correlation was observed between suprabasal expression of p53 and the presence or severity of the dysplasia. The expression of p53 is an early event in oral tumorigenesis but it does not behave as an objective marker of the presence or severity of epithelial dysplasia. PMID- 12080993 TI - Parotid neoplasms: diagnosis, treatment, and intraparotid facial nerve anatomy. AB - The demographics of parotid neoplasms in different populations have been reported by various centres. In this investigation, we reviewed retrospectively all the in patient and out-patient charts and records of 108 patients who were diagnosed with parotid neoplasms and received parotidectomies in our department from 1 January 1993 to 15 April 2000. Patient age, gender, tumour pathology, fine-needle aspiration cytology results, and the intraparotid anatomy of the facial nerve were noted. We showed that despite the difference between our Taiwanese and previously studied patient populations, both populations had a similar distribution, diagnosis and treatment of parotid neoplasms, although the incidence of parotid tuberculosis was higher in our patient group. In addition, the facial nerve anatomy within the parotid gland had three main branching patterns in the upper and lower division. PMID- 12080994 TI - Citation analysis of otorhinolaryngology journals. AB - Bibliometric analysis is used to assess the 'impact' of scientific journals. The commonest method of evaluation is impact factor. The aim of this study was to analyse the citation data for otorhinolaryngology journals of the years 1994 to 1998. Data on the total number of citations and impact factor of journals was obtained from the CD-ROM editions 1994-98 of the Journal Citation Reports and 'Web of Science' database. The adjusted impact factor and five-year impact factor has been calculated. Fifteen otorhinolaryngology journals have been identified and ranked according to the impact factor. Head and Neck has the highest adjusted impact factor. Archives of Otolaryngology - Head and Neck Surgery has the highest five-year impact factor. There is considerable variation in the ranking of journals calculated by the five-year impact factor. Impact factors of otolaryngology journals can help to direct readers to those journals that have a track record of publishing data that are frequently cited. Although there are several limitations to the use of citation data to rank journals, the authors recommend the use of the five-year period for calculation of the impact factor for ranking of otolaryngology journals. PMID- 12080995 TI - An audit of post-operative analgesia in children following tonsillectomy. AB - Pain following tonsillectomy in children is a significant problem that tends to be underestimated. We audited post-operative pain in 33 children and found significant proportions of children in moderately severe to severe pain, particularly at, or soon after returning to the ward. These findings led to changes in practice that included the administration of pre-operative paracetamol and diclofenac as well as the uniform post-operative prescription of them, with printed instructions to reduce the scope for mistakes by medical staff. Other changes included a uniform policy, with which the nursing staff could become familiar, and the regular formal recording of pain to assist in the recognition of pain and analgesic requirements. On re-auditing two years later, in a group of 100 children, post-operative pain was significantly improved. The proportion experiencing moderately severe to severe pain on returning to the ward, as their maximum pain score and pain score at discharge fell from 70 per cent to 48 per cent, 85 per cent to 56 per cent, and six per cent to two per cent respectively. PMID- 12080996 TI - Intracochlear insertion of electrodes using hyaluronic acid in cochlear implant surgery. AB - Hyaluronic acid has been proposed as a lubricant for improving the outcome of cochlear implant surgery. We describe a method which includes coating the electrodes with a thin layer of hyaluronic acid as well as using a droplet of the lubricant on the cochleostomy to provide additional magnification, like a lens, that facilitates the insertion of the electrode array. The use of hyaluronic acid reduces trauma to the cochlea and avoids contaminating perilymph by blood or by saline used during the operation. It is an efficacious and reliable method for cochlear implant surgery. PMID- 12080997 TI - Cochlear implantation in maternal inherited diabetes and deafness syndrome. AB - We describe a rare case of a gradually progressive hearing loss in a patient with maternally inherited diabetes and deafness (MIDD) syndrome successfully rehabilitated with a cochlear implant. The possible aetiology of the hearing loss in these cases is discussed. PMID- 12080999 TI - Sterile abscess mimicking recurrent tumour in the cerebellopontine angle. AB - Vestibular schwannomas are the most common tumours encountered in the cerebellopontine angle (CPA) region, accounting for 90 per cent of all lesions. Early pyogenic abscess following surgery and delayed abscesses due to retained foreign bodies have been reported. We describe a case of sterile abscess of unknown aetiology in the CPA region, occurring 13 years after surgical excision of a vestibular schwannoma. The clinical and radiological features were suggestive of recurrent vestibular schwannoma or malignant transformation. We believe this is the first reported case of delayed occurrence of sterile abscess in the CPA region. Further the diagnostic difficulties of such rare lesions occurring in the CPA after vestibular schwannoma surgery are discussed. PMID- 12081000 TI - Pneumosinus dilatans of the frontal sinuses: two cases and a discussion of its aetiology. AB - Pneumosinus dilatans is a rare condition with the dilatation of aerated paranasal sinuses. We present two cases and discuss its aetiology, the radiological classification of enlarged aerated sinuses, and its treatment. One much quoted hypothesis for the cause of this condition has been that it is due to a 'ball valve' effect of mucosal closure at the frontal recess. However, one of our subjects has started, and done, a considerable amount of subaqua diving since presenting with their condition and has had no symptoms on diving, or progression of their frontal swelling. This observation, along with the fact that retained secretions are not seen within the sinuses in this condition, raises doubt about the theory that a one-way valve is responsible. PMID- 12080998 TI - Middle cerebral artery aneurysm presenting as isolated hyperacusis. AB - We present the first case of a middle cerebral artery aneurysm presenting as isolated hyperacusis. The patient had a Guglielmi detachable coil (GDC) embolization of his aneurysm with complete resolution of his symptoms. The pathophysiological mechanism is discussed. We suggest further radiological investigation in young patients presenting with this symptom. PMID- 12081001 TI - Melkersson-Rosenthal syndrome. AB - Melkersson-Rosenthal syndrome (MRS) is a rare idiopathic non-caseating granulomatous condition. It is rarely described in otorhinolaryngology-related journals, although facial palsy, lip-swelling, and lingua plicata, are its most common presenting features. This classical triad however is not always present in MRS. Other forms of orofacial swelling exist. This paper describes a patient with a variant of MRS, treated by us with intra-lesional steroid injection. We also discuss the other differential diagnoses that may mimic MRS. PMID- 12081002 TI - Hypoglossal nerve paralysis following tonsillectomy. AB - While tonsillectomy is the commonest operation performed by otolaryngologists, paralysis of the hypoglossal nerve following tonsillectomy is not well recognized in the otolaryngology text or literature. We report a case of hypoglossal nerve paralysis following tonsillectomy and discuss the theories on the pathoaetiology as described in the predominantly anaesthetics literature. The likely causes of nerve injury are described and precautions are suggested to help avoid this problem. PMID- 12081003 TI - An unusual case of dysphagia: retained Groningen valve. AB - A Groningen speaking valve was lodged in the oesophagus in a post-laryngectomy patient and was neglected for nine years. The patient presented with dysphagia. It was diagnosed and removed on rigid endoscopy. The procedure was complicated by a primary tear of the oesophagus, that was managed conservatively. The case has many interesting features, that are discussed below. PMID- 12081004 TI - Multiple infiltrating lipomas of the tongue. AB - A rare case of multiple infiltrating lipomas in the tongue of a 54-year-old man is reported. There have been only five previously reported cases of infiltrating lipoma of the tongue. They were all solitary in nature. The present lesion is the first case reported in the English literature of multiple infiltrating lipoma of the tongue. Because of its infiltrating nature, this tumour may be mistaken for a liposarcoma. Achievement of adequate surgical margins is essential, as the recurrence rate may be as high as 62.5 per cent without complete excision. PMID- 12081005 TI - Endolymphatic sac tumours. AB - Endolymphatic sac tumours (ELST) are rare tumours of the petrous temporal bone. They may arise sporadically or be associated with von Hippel-Lindau disease. Their differential diagnosis is discussed. We present the clinical and histopathological features of two new patients with ELST and outline the management of their condition. In addition, we review a third case previously reported as a choroid plexus papilloma in which the histology has been re assessed and the diagnosis changed to ELST. The controversy regarding the cellular origins of adenomatous tumours of the temporal bone is highlighted. PMID- 12081006 TI - The JLO Travelling Fellowship 2nd report 2001. Visit to the House Ear Clinic, Los Angeles, USA and the Institut Georges Portmann, Bordeaux, France. PMID- 12081009 TI - Immune responses to myxoma virus. AB - Myxoma virus causes the systemic disease myxomatosis in the European rabbit (Oryctolagus cuniculus). Originating in the South American rabbit Sylvilagus brasiliensis, where it causes a relatively localized fibroma, myxoma virus is a classic example of a virus that has jumped species to produce an exotic disease and then coevolved with its new host. Like other poxviruses, myxoma virus encodes multiple proteins capable of downregulating the host innate and acquired immune responses. Other virus-encoded proteins enable replication in host lymphocytes and monocytes, for example, by inhibiting apoptosis. Detailed studies on these proteins have demonstrated novel methods of interactions with the host immune system and added substantially to the understanding of the interaction of large DNA viruses with their hosts. Despite the increasingly detailed molecular knowledge of myxoma virus, relatively little is known about the dynamics of the interaction of the virus with the integrated host-immune system during infection and, in particular, about the evolution of resistance to the virus in wild rabbits or the species barrier. This review will focus on the detailed molecular studies that have been done with myxoma virus and discuss the more limited knowledge of the pathogenesis of myxoma virus in rabbits and the ways that the consolidated immune responses may determine genetic resistance to myxomatosis. PMID- 12081007 TI - Guidelines for preventing opportunistic infections among HIV-infected persons- 2002. Recommendations of the U.S. Public Health Service and the Infectious Diseases Society of America. AB - In 1995, the U.S. Public Health Service (USPHS) and the Infectious Diseases Society of America (IDSA) developed guidelines for preventing opportunistic infections (OIs) among persons infected with human immunodeficiency virus (HIV); these guidelines were updated in 1997 and 1999. This fourth edition of the guidelines, made available on the Internet in 2001, is intended for clinicians and other health-care providers who care for HIV-infected persons. The goal of these guidelines is to provide evidence-based guidelines for preventing OIs among HIV-infected adults and adolescents, including pregnant women, and HIV-exposed or infected children. Nineteen OIs, or groups of OIs, are addressed, and recommendations are included for preventing exposure to opportunistic pathogens, preventing first episodes of disease by chemoprophylaxis or vaccination (primary prophylaxis), and preventing disease recurrence (secondary prophylaxis). Major changes since the last edition of the guidelines include 1) updated recommendations for discontinuing primary and secondary OI prophylaxis among persons whose CD4+ T lymphocyte counts have increased in response to antiretroviral therapy; 2) emphasis on screening all HIV-infected persons for infection with hepatitis C virus; 3) new information regarding transmission of human herpesvirus 8 infection; 4) new information regarding drug interactions, chiefly related to rifamycins and antiretroviral drugs; and 5) revised recommendations for immunizing HIV-infected adults and adolescents and HIV exposed or infected children. PMID- 12081010 TI - Effects of interferon-gamma on neuronal infections. AB - IFN-gamma is released in the brain, not only during acute infectious diseases or immunological reactions, but also for extended periods of time after clearance of infectious virus and during viral latency. In this review, we focus on the role of IFN-gamma in the interaction between a neuron and a pathogen, that is, the role in implementation of microbial destruction, stasis or persistence, its potential neuroprotective or toxic effects, and how this cytokine may affect neuronal function and the behavior of an individual. PMID- 12081011 TI - Mouse hepatitis virus infection of the central nervous system: chemokine-mediated regulation of host defense and disease. AB - Infection of the central nervous system (CNS) of susceptible mice with mouse hepatitis virus (MHV), a positive-strand RNA virus that is a member of the Coronaviridae family, reproducibly results in an acute encephalomyelitis followed by a demyelinating disease similar to the human demyelinating disease multiple sclerosis (MS). MHV infection triggers a robust cell-mediated response in which both CD4+ and CD8+ T cells are essential in controlling viral replication and spread. However, viral clearance is incomplete and viral RNA and protein can persist within white matter tracts, areas of viral persistence are often associated with demyelinating lesions, and recent studies have indicated an important role for both T cells and macrophages in contributing to myelin destruction. The molecular mechanisms governing leukocyte trafficking and accumulation within the CNS of MHV-infected mice are just now being understood and recent studies indicate that chemokines and chemokine receptors have an important role in this process. This article will provide an overview on how these molecules regulate T cell and macrophage trafficking into the CNS of MHV infected mice and illustrate the delicate balance that exists with regards to expression of chemokines and their receptors as it relates to both host defense and disease development. PMID- 12081012 TI - Regulation of immune recognition molecules by flavivirus, West Nile. AB - We have shown the flaviviruses can up-regulate the cell surface expression of the immune recognition molecules, major histocompatability complex class-I and class II (MHC-I, MHC-II), ICAM-1, VCAM, and E-selectin, in an interferon-independent and tumor necrosis factor-independent manner. This up-regulation is associated with an increased transcription of the relevant genes and is due to activation of the transcription factor, nuclear factor-kappa B. The level of up-regulation is determined in part by the cell cycle position of the cell when infected with the flavivirus, as quiescent cells show a greater increase in the level of expression of the immune recognition molecules, MHC-I and ICAM-1, than cells in other phases of the cell cycle. The resultant increased cell surface expression is functional with the increased expression resulting in increased recognition by flavivirus specific and allo-specific cytotoxic T cells. PMID- 12081013 TI - T cell failure in hepatitis C virus infection. PMID- 12081014 TI - Principles underlying the development and use of live attenuated cold-adapted influenza A and B virus vaccines. PMID- 12081015 TI - A longitudinal study of plasma and salivary antibodies in HIV-1 infection. AB - The patterns of plasma and salivary IgG and IgA antibodies reacting to HIV-1 proteins were followed in seven HIV-1-infected individuals for a period of 18-40 months. Western blot analyses revealed diversities in specificity of these antibodies among subjects; however, for the same subject, the specificity profile remained consistent throughout the entire follow-up period. The staining intensities of plasma IgG from two subjects were associated with plasma viral load. The band intensities of salivary IgG were mostly determined by plasma IgG; the health of the oral cavity might also influence the transudation of salivary IgG antibodies. The binding intensities of plasma and salivary IgA antibodies specific for certain viral proteins were associated with plasma viral load in some subjects as well. PMID- 12081016 TI - Induction of a strong HIV-specific CD8+ T cell response in mice using a fowlpox virus vector expressing an HIV-1 multi-CTL-epitope polypeptide. AB - Recombinant avipoxvirus vectors are attractive candidates for use in vaccination strategies for infections such as human immunodeficiency virus type 1 (HIV-1), where induction of a CD8+ T cell response is thought to be an important component of protective immunity. Here, we report the expression of a multiepitope polypeptide (TAB9) composed of the central 15 amino acids of the V3 loop from six different isolates of HIV-1 in a fowlpox virus (FWPV) vector, and the use of this vector (FPTAB9LZ) to induce strong HIV-specific CD8+ T cell responses in mice. In animals immunized twice intravenously with FPTAB9LZ, almost 2% of the CD8+ T cells in the spleen were shown to produce IFN-gamma in response to stimulation with HIV-1 peptides 1 week after the second immunization. The most dominant response was to the HIV-1 IIIB peptide. A strong HIV-specific response was also induced by intraperitoneal immunization of mice with FPTAB9LZ, whilst subcutaneous immunization elicited a weaker response. Intraperitoneal immunization with FPTAB9LZ was also shown to provide protection against challenge with a recombinant vaccinia virus expressing antigens, including those in TAB9. These results confirm the potential of FWPV vectors for use in HIV vaccination strategies. PMID- 12081017 TI - Respiratory syncytial virus and pulmonary surfactant. AB - Respiratory syncytial virus (RSV) infections peak in young infants and are associated with significant morbidity. The collectins surfactant protein-A (SP-A) and SP-D are pattern recognition molecules that belong to the innate immune system of the lungs, forming a first line of defense. On the one hand, SP-A and SP-D levels are reduced during RSV infection. This may critically influence the invasion of RSV and also the virus-induced cytokine patterns of the host. Both collectins enhance the in vivo elimination of RSV. Thus, interactions before the virus enters the epithelial cells may determine the course of the infection. On the other hand, during severe RSV infection in infants, the biophysical surfactant function is reduced and exogenous surfactant substitution may be a valid therapeutic option for selected infants. Thus, all components of the pulmonary surfactant system are involved during severe RSV infection. Especially the collectins SP-A and SP-D may play a pivotal role determining the short- and long-term course of RSV infections in early infancy. PMID- 12081018 TI - Innate immunity in viral encephalitis: role of C5. AB - The complement system is a critical component of both the innate and acquired immune systems. It is important in host defense against viruses, bacteria, and fungi for opsonization and for lysis of pathogens. However, activated complement can also cause tissue damage. There is compelling evidence that complement factors are presented in the central nervous system (CNS). Complement activation (by any of the three pathways: classical, alternate, and lectin) can lead to inflammation and tissue damage, while at the same time may also restrict certain pathogens in the CNS. C5a is formed by proteolytic cleavage C5. C5a is considered the most potent proinflammatory mediator, often called an anaphylotoxin. In this communication, we examine the roles of C5 (C5a) in vesicular stomatitis virus (VSV)-induced encephalitis. We found that C5a is produced during VSV infection, but C5-deficient mice had similar pathology as their controls. We concluded that C5 is not a critical factor in mediating the host response during VSV encephalitis. PMID- 12081019 TI - A longitudinal study of cell-mediated immunity in pigs infected with porcine parvovirus. AB - Porcine parvovirus (PPV) is an ubiquitous pathogen causing reproductive failure in swine. Protection against reproductive failure caused by acute PPV infection has commonly been related to the presence of specific antibodies in the dam. However, the role of cell-mediated immunity during chronic PPV infection remains to be elucidated, and may be relevant to the pathogenesis of novel diseases such as postweaning multisystemic wasting syndrome (PMWS), which may be triggered by coinfection with PPV and porcine circovirus type 2 (PCV2). To investigate whether pigs infected with PPV generate a cell-mediated immune response, a longitudinal infection experiment was performed, using swine leukocyte antigens (SLA) class I characterized growing pigs (haplotype H7/H7). Pigs were intranasally inoculated with PPV at 0, 80, and 136 days. At predetermined time points, peripheral blood mononuclear cells (PBMC) were isolated, and virus-specific lymphoproliferative responses and the cytolytic activities of cytotoxic T-lymphocytes (CTL) and natural killer (NK) cells were examined. Cytolytic assays were performed by the chromium release method, using as targets a syngeneic porcine kidney cell line established for the purpose (CTL assays) and K562 cells (NK assays). A specific proliferative response of PBMC from virus-infected pigs to PPV was observed from day 101 onwards. In contrast, PBMC from mock-infected pigs did not proliferate in response to PPV. Flow cytometric analysis indicated that the CD4+CD8+ T-cell subset of PBMC proliferated in response to virus antigen, in keeping with the assumed role for these cells in immunological memory. This is, to our knowledge, the first indication of a cellular immune response following PPV infection. A weak CTL activity, which peaked on days 80 and 87, was observed in PPV-infected pigs. In vitro restimulation of PBMC with live PPV did not induce further CTL activity. A pronounced NK cell activity was detected in both virus-infected and control pigs throughout the experiment, and may have negatively affected the sensitivity of the CTL assay. In conclusion, the findings of a late lymphoproliferative response together with weak CTL activity are in keeping with an effective control of acute PPV infection by humoral immunity, but open the possibility that cellular immunity may play a role in controlling PPV reinfection. Finally, we find that the established experimental model using SLA characterized pigs may constitute a valuable tool for future studies of CTL activity in pigs. PMID- 12081020 TI - BHV-1-Specific CD4+, CD8+, and gammadelta T cells in calves vaccinated with one dose of a modified live BHV-1 vaccine. AB - Expression of the high-affinity interleukin 2 receptor alpha chain (CD25) was used to monitor antigen-specific activation of T lymphocyte subsets (CD4+, CD8+, and gammadelta T cells) from cattle immunized with modified live bovine herpesvirus-1 (BHV-1). Calves seronegative for BHV-1 were either vaccinated with one dose of modified live vaccine containing BHV-1 or not vaccinated to serve as negative controls. Two animals vaccinated 7 and 5 weeks before the start of the experiment with two doses of modified live vaccine containing BHV-1 served as positive controls. Blood samples were taken from the nonvaccinate group, the positive control group, and the vaccinate group at 0, 21, 35, 60, and 90 days postinoculation (PI). Isolated peripheral blood mononuclear cells from immunized and control animals were incubated for 5 days with and without live BHV-1 ISU99. Compared to the nonvaccinates, a significant (p < 0.05) increase in expression of CD25 by CD4+, CD8+, and gammadelta T lymphocytes from the vaccinate group was detected following in vitro exposure to live BHV-1 after vaccination. This is apparently the first report using live BHV-1 to stimulate lymphocytes in vitro and showing CD8+ T cell activation. Peripheral blood from the positive control animals was depleted of CD4+, CD8+, or gammadelta T lymphocytes prior to incubation with BHV-1 to assess bystander activation in the CD25 expression assay. When incubated with live BHV-1, depletion of CD4+ T cells depressed the expression of CD25 by CD8+ T cells, but not gammadelta T cells. Depleting CD8+ or gammadelta T cells prior to in vitro culture with BHV-1 did not affect the expression of CD25 by the remaining T lymphocyte subsets. Vaccinates were protected from challenge with virulent BHV-1 at 110 days postvaccination compared to nonvaccinates. Expression of CD25 appears to be a useful marker for evaluating induction of antigen-specific T lymphocyte subset responses following vaccination. PMID- 12081021 TI - Effect of ultraviolet-absorbing vinyl film on organophosphorus insecticides dichlorvos and fenitrothion residues in spinach. AB - Dichlorvos and fenitrothion residues found in spinach grown in greenhouse covered either by regular vinyl film or UV-absorbing vinyl film (UV-A) were analyzed by gas chromatography. After one day, dichlorvos residues in spinach covered with regular vinyl film and UV-A degraded by 97% and 80%, respectively, and degraded 100% after three days covered with regular vinyl film and six days covered with UV-A. After three days, fenitrothion residue in spinach covered with regular vinyl film degraded by 72% and then by 97% after six days; residue in spinach covered with UV-A degraded by 50% after three days and by 95% after six days. These results indicate that UV-A used to prevent the occurrence of insects and fungi in greenhouses reduced the degradation rates of dichlorvos and fenitrothion. PMID- 12081022 TI - Bioavailability and biodegradation of prosulfocarb in soil. AB - Active microbial degraders of the herbicide prosulfocarb (PSC) were isolated to evaluate their performance in soil with a view to their use for bioremediation. The isolated cultures (a microbial consortium and a Pseudomonas sp. strain) were active when tested in mineral medium with PSC as the only carbon source, but had an adverse effect on the soil indigenous microflora. Biodegradation in the inoculated soils was thus lower than in the uninoculated soil when only the indigenous microflora was present. Further tests showed that the strong affinity of PSC for soil organic matter affected its bioavailability and hence its biodegradation by the inocula. Bioremediation of PSC contaminated soils could thus be undertaken by biostimulation of indigenous microflora. PMID- 12081023 TI - Development of ELISA technique for the analysis of atrazine residues in water. AB - A highly sensitive enzyme immunoassay is described for the detection of atrazine residues in water. Atrazine derivative was conjugated to Bovine Serum Albumin (BSA) to obtain an immunizing antigen and to Horseradish Peroxidase enzyme (POD) to obtain a marker for immunoassay. The formation of these conjugations was confirmed by UV spectroscopy as well as by gel-electrophoresis. Polyclonal antibodies were raised in rabbits by immunization with an atrazine-BSA conjugate containing 29 atrazine residues per BSA molecule. An ELISA on microtitration plates was optimized with peroxidase-atrazine conjugate. The middle of the test (50% B/Bo) was found to be at 90 ng/l, which is well below the maximum concentration permitted by the EC guidelines for drinking water. Detection limits for atrazine of about 1 ng/l could be reached. The assay did not require concentration or cleanup steps for drinking or ground water samples. Validation experiments showed good accuracy and precision. No cross-reactivities were shown by other s-triazines like terbutryn, ametryn, terbuthylazine, des isopropylatrazine, and de-ethylatrazine except hydroxyatrazine. The latter was present at very low levels that can be calibrated/standardized before analysis or it may be considered as leftover residues of atrazine. Based on these results, it is suggested that this test can be applied to obtain fairly accurate results for atrazine concentration in water samples from different sources. PMID- 12081024 TI - Pesticide residues in cow milk consumed in Sao Paulo City (Brazil). AB - Residues of pesticides were surveyed in 132 samples of cow milk collected from bulk transports (38 samples of raw milk) and market (94 samples of pasteurised milk). These samples were analysed by the multiresidue analytical method DFG S19 for pesticide contamination. More than 70 active substances were studied and the identification and quantification were made by gas chromatographic technique. The results showed that 0.76% of samples were contaminated with HCH (alpha isomer) and 10.60% with endosulfan (alpha and beta isomers). Both pesticides, endosulfan and HCH, found in milk samples, indicated their use in agriculture practices, although legislation in Brazil does not permit the use of HCH since 1985 and endosulfan can be used only in a few crops. These compounds should not be present in milk because there are implications on human health. Organophosphorus, carbamates, pyrethroids, herbicides, and fungicides were not detected in cow's milk samples. PMID- 12081025 TI - The effects of surfactants on adhesion, spreading, and retention of herbicide droplet on the surface of the leaves and seeds. AB - Droplet adhesion, spreading, and retention on leaf and seed surfaces are studied in the presence of surfactants for successful formulation of a herbicide whereby efficient coverage, less wastage, and environment protection are achieved. Cationic, anionic, and nonionic surfactants, e.g., HTAB, SDBS, and tergitol, were added to water and the static contact angle of water droplet was measured on cauliflower, cabbage, spinach leaves, French bean, wheat, and Bengal gram seeds. Droplet adhesion, spreading, and retention were analysed in terms of static contact angle and physical properties of the system. The presence of a small quantity (0.1 wt.%) of tergitol in water increased the adhesion, spreading, and retention of droplet on all leaf and seed surfaces tested. Tergitol, a nonionic surfactant, is likely to be adsorbed more than the ionic surfactants HTAB and SDBS at the water/leaf (or seed)/air interface since plant leaves and seeds are nonionic in nature. The addition of 0.1% wt. of tergitol to a common herbicide, derosal, increases the adhesion, spreading, and retention of derosal on leaf and seed surfaces, consequently, efficient coverage and minimum wastage are attained. PMID- 12081026 TI - Persistence and bioactivity of metsulfuron-methyl in three soils. AB - The persistence of metsulfuron-methyl (methyl 2-[[(4-methoxy-6-methyl-1,3,5 triazin-2-yl)aminocarbonyl]aminosul fonyl]benzoate) in nonautoclaved and autoclaved Selangor, Lating, and Serdang series soils incubated at different temperatures and with different moisture contents was investigated under laboratory conditions using cucumber (Cucumis sativus L.) as the bioassay species. Significant degradation of metsulfuron-methyl was observed in nonautoclaved soil compared with the autoclaved soil sample, indicating the importance of microorganisms in the breakdown process. At higher temperatures the degradation rate in nonautoclaved soil improved with increasing soil moisture content. In nonautoclaved Selangor, Lating and Serdang series soils, the half life was reduced from 4.79 to 2.78 days, 4.9 to 3.5, and from 3.3 to 1.9 days, respectively, when the temperature was increased from 20 degrees to 30 degrees C at 80% field capacity. Similarly, in nonautoclaved soil, the half-life decreased with an increasing soil moisture from 20% to 80% at 30 degrees C in the three soils studied. In the autoclaved soil, the half-life values were slightly higher than those obtained in the nonautoclaved soils, perhaps indicating that the compound may be broken down by nonbiological processes. The fresh weight of the bioassay species was reduced significantly in Serdang series soil treated with metsulfuron-methyl at 0.1 ppm. However, the reduction in fresh weight of the seedlings was least in Lating series soil, followed by Selangor series soil. PMID- 12081027 TI - Adsorption, desorption, and mobility of two insecticides in Malaysian agricultural soil. AB - Laboratory studies utilizing radioisotopic techniques were conducted to determine the adsorption, desorption, and mobility of endosulfan (6,7,8,9,10,10-hexachloro 1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxanthiepin3-oxide) and methamidophos (O,S-dimethyl phosphorothioate) in sandy loam and clay soils of the Cameron Highlands and the Muda rice-growing area, respectively. High Freundlich adsorption distribution coefficients [Kads(f)] for endosulfan (6.74 and 18.75) and low values for methamidophos (0.40 and 0.98) were obtained in the sandy loam and clay soils, respectively. The observed Koc values for endosulfan were 350.85 (sandy loam) and 1143.19 (clay) while Koc values of 20.92 (sandy loam) and 59.63 (clay) were obtained for methamidophos. Log Kow of 0.40 and 1.25 were calculated for endosulfan as well as -1.96 and -1.21 for methamidophos in the sandy loam and clay soils, respectively. Desorption was common to both pesticides but the desorption capacity of methamidophos from each soil type far exceeded that of endosulfan. Soil thin layer chromatography (TLC) and column studies showed that while methamidophos was very mobile in both soils, endosulfan displayed zero mobility in clay soil. PMID- 12081028 TI - Dynamic air sampling of volatile organic compounds using solid phase microextraction. AB - A new dynamic air sampling system was devised and evaluated in conjunction with solid phase microextraction (SPME) fiber materials for extracting odor-causing volatile organic compounds (VOCs) present in swine building environments. Utilizing a standard solution consisting of 11 compounds (i.e., volatile fatty acids, indoles, and phenol), sampling times, volumes, and flow rates were adjusted to establish optimal extraction conditions. Results indicated that the sampling system was effective with the Carboxen/Polydimethylsiloxane (CAR/PDMS) fiber in extracting all 11 standard compounds. The best sampling conditions for the extraction were a 100-mL sampling vial subjected to a continuous flow of 100 mL/min for 60 min. The gas chromatographic analysis showed that the reproducibility was within acceptable ranges for all compounds (RSD=4.24-17.26% by peak areas). In addition, field tests revealed that the sampling system was capable of detecting over 60 VOCs in a swine house whose major components were identified by gas chromatography-mass spectrometry (GC-MS) and by their retention times as volatile fatty acids, phenols, indole, and skatole. The field tests also showed that considerably different levels of VOCs were present in various parts of the swine building. PMID- 12081029 TI - Sequence analysis and molecular characterization of a nitrocatechol dioxygenase gene from Pseudomonas putida. AB - A Pseudomonas putida capable of degrading polychlorinated biphenyl was also found to transform 4-nitrocatechol to 3-nitro-2-hydroxy-6-oxohexa-2,4-dienoic acid (NHODA). Crude cell extract of this bacterium exhibited an enzyme (nitrocatechol dioxygenase, Ndo) activity catalyzing this transformation. The gene encoding Ndo was cloned in E. coli. The cloned gene (ndo) expressed in E. coli had enzyme activity that degraded not only 4-nitrocatechol but also 4-chlorocatechol, 4 methylcatechol, 2,3-dihydroxybiphenyl, and 4'-chloro-2,3-dihydroxybiphenyl. Nucleotide sequence analysis of the cloned ndo exhibited an open reading frame of 939 base pairs. This sequence can encode a 313 amino acids protein of approximately molecular weight of 35 kd, which was confirmed by in vitro transcription and translation assay and SDS-PAGE analysis. A putative ribosomal binding site (GAGGAGA) was present 7 base pairs upstream from the AUG start codon and a promotor site homologous to E. coli '-10' and '-35' regulatory region was located at '-123' and '-174' area of our clone with sequences of TTGAAG and GTGACA, respectively. The deduced amino acid sequence showed 69% homology with Cdo from Burkholderia cepacia AAI. A unique insertion of 21 amino acids was found towards the N-terminal of the Ndo. Expression of ndo in strain OU83 was repressed in presence of 3-chlorobenzoic acid as judged by the decrease in the expression of ndo specific transcript. PMID- 12081030 TI - Detection of infectious and toxigenic bacteria. PMID- 12081031 TI - Metabolic profiling of rodent biological fluids via 1H NMR spectroscopy using a 1 mm microlitre probe. AB - The application of a 1 mm TXI (1H/13C/15N) microlitre NMR probe with z-gradient for metabolic profiling of biofluids is described. The probe was used to provide spectral profiles for rat blood plasma using only approximately 2 microl of fluid with a range of solvent suppression techniques. Using a similar amount of fluid, spectra were obtained from rat and mouse cerebrospinal fluid, demonstrating that the probe could be used to profile rodents metabolically via biofluids previously inaccessible to NMR analysis without the need for termination. PMID- 12081032 TI - Construction and control of plasmid DNA network. AB - The influences of different cations on plasmid DNA network structures on a mica substrate were investigated by atomic force microscopy (AFM). Interactions between the DNA strands and mica substrate, and between the DNA strands themselves were more strongly influenced by the complex cations (Fe(phen)3(2+), Ni(phen)3(2+), and Co(phen)3(3+)) than by the simple cations (Mg2+, Mn2+, Ni2+, Ca2+, Co3+). The mesh height of the plasmid DNA network was higher when the complex cations were added to DNA samples. The mesh size decreased with increasing DNA concentration and increased with decreasing DNA concentration in the same cation solution sample. Hence, plasmid DNA network height can be controlled by selecting different cations, and the mesh size can be controlled by adjusting plasmid DNA concentration. PMID- 12081033 TI - Using flow injection analysis to time-resolve rhythmic and pulsatile signals. AB - Continuous monitoring can be used to detect rhythms, an important aspect of biology. But peaks of concentration are broadened by dispersion so that they overlap their neighbours and obscure high frequency chemoperiodicities. In this study, flow injection was found experimentally to be useful in resolving these. A rhythmically varying pattern of permanganate concentration was measured spectrophotometrically. The rhythm (frequency 0.08 Hz) was observable at a dispersion coefficient of 3.0 but not at 3.9 (when only a single peak was recorded). It was again observable using the same high dispersion manifold but positioned after an injection valve that subsampled the stream at intervals. A design based on this work is proposed for an automated instrument that outputs a time series of concentration measures. PMID- 12081034 TI - Electrochemical solid-phase microextraction of anions and cations using polypyrrole coatings and an integrated three-electrode device. AB - A method for the extraction, transfer and desorption of anions and cations under controlled potential conditions employing a new integrated three-electrode device is described. The device, containing working, reference and counter electrodes, was prepared from tubes that could be moved vertically with respect to each other. In this way, a small amount of solvent, held by capillary force, remained between the electrodes when the device was lifted out of a solution after an extraction. This design allowed the potential control to be maintained at all times. With the new integrated device, it was possible to perform potential controlled desorption into vials containing as little as 200 microl of solution. The required ion exchange capacity was obtained by electrodeposition of a polypyrrole coating on the surface of the glassy carbon working electrode. Solid phase microextractions of several cations or anions were performed simultaneously under potentiostatic control by doping the polypyrrole coating with different anions such as perchlorate and p-toluenesulfonate. The efficiency of the extractions, which could be altered by varying the potential of the working electrode, could be increased by 150 to 200% compared to extractions using normal solid-phase microextraction conditions under open circuit conditions. A constant potential of +1.0 V and -0.5 V with respect to the silver pseudo reference electrode, was found to be well-suited for the extraction of samples containing ppm concentrations of anions (chloride, nitrite, bromide, nitrate, sulfate and phosphate) and cations (cadmium, cobalt and zinc), respectively. PMID- 12081035 TI - On-fiber derivatization for direct immersion solid-phase microextraction. Part II. Acylation of amphetamine with pentafluorobenzoyl chloride for urine analysis. AB - On-fiber derivatization was used for solid-phase microextraction (SPME) in order to increase the detectability and extractability of drugs in biological samples. Amphetamine, which was used as a model compound, was derivatized with pentafluorobenzoyl chloride (PFBCl) and subjected to gas chromatography with electron capture or mass spectrometric detection. Extraction was performed by direct immersion of a 100 microm polydimethylsiloxane-coated fiber into buffered human urine. On-fiber derivatization was performed either after or simultaneously with extraction. The former procedure gave cleaner chromatograms but the latter turned out to be superior with respect to linearity and repeatability. For the on fiber derivatization of amphetamine an excess of reagent is required. Because a considerable part of the PFBCl loaded on to the fiber is used up by reaction with matrix compounds and water, a reagent loading time of 5 min was needed to obtain a linear range (r = 0.9756) from 250 pg mL(-1) to 15 ng mL(-1). Due to an interfering matrix compound, the limit of detection was also found to be dependent on the reagent loading time, i.e., the limit of detection for a PFBCl loading time of 5 min is 250 pg mL(-1) whereas that for a 1 min loading time it is 100 pg mL(-1). The relative standard deviation (n = 7) of the method was about 11% at an amphetamine concentration of 1 ng mL(-1). The applicability of the method for the determination of drugs in biological samples is shown. PMID- 12081036 TI - A new procedure for solid-phase microextraction using a flow-through system with air segmentation. AB - Three different procedures for solid-phase microextraction (SPME) of trace organic compounds from liquids were evaluated: simple extraction with magnetic stirring, use of a laboratory-made flow-through extraction cell and a new system based on a laboratory-made flow-through extraction cell in which the sample stream is segmented with air bubbles. In this new procedure, the sample flows through the cell as a regular sequence of small plugs separated by air bubbles instead of in a continuous stream. This new system combines the advantages of SPME, as a simple, fast, sensitive and solvent-free sample concentration/introduction technique, with the advantages of on-line processing of aqueous sample as a less time-consuming, efficient and continuous technique. For testing the new flow-through procedure, a mixture of phthalate esters was used for direct absorption from an aqueous solution with a 100 microm polydimethylsiloxane fiber. The efficiency and precision of the procedure were compared with those of extraction with magnetic stirring and the usual flow through cell concept. The results indicate better contact of the analytes with the extraction film when the air-segmented flow-through extraction cell is employed. PMID- 12081037 TI - Liquid-phase microextraction of protein-bound drugs under non-equilibrium conditions. AB - Recently, we introduced an inexpensive and disposable hollow fiber-based device for liquid-phase microextraction (LPME) where ionic analytes typically were extracted and preconcentrated from 1-4 mL aqueous samples (such as plasma and urine) through an organic solvent immobilized in the pores of a polypropylene hollow fiber and into a 10-25 microL volume of acceptor phase present inside the lumen of the hollow fiber. Subsequently, the acceptor phase was directly subjected to the final analysis by a chromatographic or electrophoretic method. In the present work, attention was focused on LPME of the basic drugs amphetamine, pethidine, promethazine, methadone and haloperidol characterized by substantial differences in the degree of protein binding. Drug-protein interactions in plasma resulted in reduced recoveries and substantially increased extraction times compared with extraction of the drugs from a pure water matrix. However, by addition of 5-50% methanol to the plasma samples, recoveries were comparable with LPME from water samples and ranged between 75 and 100%. The addition of methanol was found not to speed up the LPME process and extractions from plasma were performed in 45 min to reach equilibrium. Because approximately 55-70% of the final analyte concentrations were achieved within the initial 10 min of the LPME process, validation was accomplished after 10 and 45 min of LPME. In general, the results with 10 and 45 min were almost comparable, with precision data in the range 1.2-11.1% (RSD) and with linearity in the concentration range 20-1000 ng mL(-1) (r = 0.999). In conclusion, excellent LPME results may be achieved in a short time under non-equilibrium conditions with a minor loss of sensitivity. In cases of drug-protein interactions, methanol may be added to ensure a high extraction recovery. PMID- 12081038 TI - Measurement of carbonaceous aerosols: validation and comparison of a solvent extraction-gas chromatographic method and a thermal optical transmittance method. AB - A solvent extraction-gas chromatographic method was developed for the measurement of four classes of solvent-extractable organic compounds (SEOC), viz., aliphatic hydrocarbons, polynuclear aromatic hydrocarbons, fatty acids and alkanols, in ambient carbonaceous aerosols. The selectivity and recovery of the method using different solvents commonly used in the analysis of SEOC was evaluated. The composition of SEOC in aerosol and urban dust samples was found to vary with the choice of solvent. Such variation was not observed in filters spiked with standards of SEOC. The developed method was validated by the analysis of reference urban dust SRM 1649a from the National Institute of Standards and Technology. The concentrations of selected polynuclear aromatic hydrocarbons in the reference dust were found to fall within the certified and reference concentrations. The thermal characteristics of aerosol samples collected in an urban area of Hong Kong and the four classes of identified SEOC were also studied using a thermal optical transmittance (TOT) method as specified in NIOSH Method 5040. Correlations were made to compare the method-dependent SEOC content obtained by the solvent extraction method and the results for total carbon, organic carbon and elemental carbon obtained by the TOT method. PMID- 12081039 TI - Inner chromatogram projection (ICP) for resolution of GC-MS data with embedded chromatographic peaks. AB - The chromatographic peak located inside another peak in the time direction is called an embedded or inner peak in contradistinction with the embedding peak, which is called an outer peak. The chemical components corresponding to inner and outer peaks are called inner and outer components, respectively. This special case of co-eluting chromatograms was investigated using chemometric approaches taking GC-MS as an example. A novel method, named inner chromatogram projection (ICP), for resolution of GC-MS data with embedded chromatographic peaks is derived. Orthogonal projection resolution is first utilized to obtain the chromatographic profile of the inner component. Projection of the two-way data matrix columnwise-normalized along the time direction to the normalized profile of the inner component found is subsequently performed to find the selective m/z points, if they exist, which represent the chromatogram of the outer component by itself. With the profiles obtained, the mass spectra can easily be found by means of a least-squares procedure. The results for both simulated data and real samples demonstrate that the proposed method is capable of achieving satisfactory resolution performance not affected by the shapes of chromatograms and the relative positions of the components involved. PMID- 12081040 TI - A novel approach for speciation of airborne chromium by convective-interaction media fast-monolithic chromatography with electrothermal atomic-absorption spectrometric detection. AB - A new analytical procedure using an anion-exchange separation support based on convective-interaction media (CIM) was developed for the speciation of chromium. The separation of Cr(VI) was performed on a weak anion-exchange CIM diethylamine (DEAE) fast-monolithic chromatographic disc. Buffer A (0.005 mol dm(-3) TRIS-HCl, pH 8.0) and buffer B (buffer A plus 3 mol dm(-3) NH4NO3) were employed in the separation procedure. The separated chromium species were determined 'off-line' by ETAAS in 0.5 cm3 fractions. The applicability of the CIM DEAE-ETAAS procedure was investigated for the determination of airborne Cr(VI) at a plasma cutting workplace. Aerosols were collected on polycarbonate membrane filters of 8 and 0.4 microm pore size (inhalable and respirable aerosols). Alkaline extraction of filters in a heated ultrasonic bath was applied to leach chromium. Good repeatability of measurement (+/-3.0%) of the alkaline extracts was obtained for Cr(VI). The LOD (3s) was found to be 0.30 microg m(-3) Cr(VI), when 0.25 m3 of air was collected on the filter. The validation of the procedure was performed by spiking filters with Cr(VI) and by the analysis of the standard reference material CRM 545, Cr(VI) in welding dust loaded on a filter. Good recoveries for spiked samples (101-102%) and good agreement between Cr(VI) found and the reported certified value for CRM 545 were obtained. The extracts were also analysed by the FPLC-ETAAS technique. Good agreement between two techniques (r2 = 0.9978) confirmed the reliability of the CIM DEAE-ETAAS procedure developed. The main advantage of the procedure lies in the speed of the chromatographic separation (chromatographic run completed in 15 min). PMID- 12081041 TI - Arsenic speciation in chinese seaweeds using HPLC-ICP-MS and HPLC-ES-MS. AB - Three common Chinese edible seaweeds, one brown (Laminaria japonica) and two red (Porphyra crispata and Eucheuma denticulatum), were examined for their total arsenic content. The As species were extracted with yields of 76.4, 69.8 and 25.0%, respectively. Anion-exchange and cation-exchange high-performance liquid chromatography (HPLC) in combination with inductively coupled plasma mass spectrometry (ICP-MS) were used for the separation of the different arsenic species in two of the three seaweed extracts (Laminaria and Porphyra). The main arsenic species in the algal extracts are arseno sugars, although it has been shown that the Laminaria seaweed contains significant amounts of dimethylarsinic acid (DMA). HPLC was coupled with electrospray mass spectrometry (ES-MS) for structural confirmation of the arsenic species. The mass spectrometer settings for the arseno sugars were optimised using standards. The conclusions drawn on the basis of HPLC-ICP-MS were confirmed by the HPLC-ES-MS data. The HPLC-ES-MS method is capable of determining both arseno sugars and DMA in the seaweeds. The unknown compounds seen in the HPLC-ICP-MS chromatogram of Laminaria could not be ascribed to trimethylarsenic oxide or tetramethylarsonium ion. PMID- 12081042 TI - Liquid chromatography-electrospray mass spectrometric studies of ginkgolides and bilobalide using simultaneous monitoring of proton, ammonium and sodium adducts. AB - A liquid chromatographic-electrospray mass spectrometric method was developed for the determination of ginkgolides and bilobalide and was applied to the analysis of commercial products of Ginkgo biloba leaf extracts. Adducts of these compounds with ammonium, proton and sodium were detected and their relative abundance depended on the electrospray fragmentor voltage. The relative standard deviation (RSD) was improved from > 17% to < 6%, when three adduct ions of (M + H)+, (M + NH4)+ and (M + Na)+ were used for quantification compared with single ion monitoring. The characteristic mass spectra of bilobalide were different from those of ginkgolides; in particular, dimers of (2M + Na)+ were observed for bilobalide only. Analysis of 26 commercial ginkgo products revealed large variations in the composition and concentrations of ginkgolides and bilobalide in herbal products. PMID- 12081043 TI - Amperometric detection of sulfur-containing compounds in alkaline media. AB - A copper-based chemically modified glassy carbon electrode (GC-Cu) was developed to be used as an amperometric sensor for electrochemically detecting several sulfur-containing compounds in alkaline media. Under optimised flow injection conditions the calibration curves for sulfite, sulfide, thiosulfate, cysteine, cystine, etc., were linear over three orders of magnitude of concentration. Detection limits were of the order of 0.04-1.5 microM. A simple and rapid method for determining sulfite in red and white wines by anion-exchange chromatography with electrochemical detection is described. PMID- 12081044 TI - Carbon nanotube-modified electrodes for the simultaneous determination of dopamine and ascorbic acid. AB - The voltammetric separation of dopamine and ascorbic acid was studied with cyclic voltammetry at two kinds of carbon nanotube-modified electrodes (coated and intercalated). The anodic peak difference reached 270 mV under the present conditions. The separation mechanism and effect factors were carefully studied. Using various types of surfactants as coating dispersants of carbon nanotubes, it was demonstrated that the charge nature of the surfactants had a strong effect on the electrochemical behavior of dopamine and ascorbic acid. When the oxidation solution of carbon nanotubes was changed from the most commonly used mixed concentrated nitric acid and sulfuric acid (1 + 3 v/v) to dilute nitric acid and to hydrochloric acid, the anodic peak separation value of dopamine and ascorbic acid increased significantly, and it was shown that carboxylic acid groups attached to the carbon nanotubes were an adverse factor for the discrimination of DA from AA. These results indicated that the resolution of DA and AA was mainly attributable to the stereo porous interfacial layer formed from aggregated pores and inner cavities of the carbon nanotubes. The modified electrodes exhibited an attractive ability to measure DA and AA simultaneously and showed good stability and reproducibility. PMID- 12081045 TI - Estimation of second order rate constants using chemometric methods with kinetic constraints. AB - Several methods are described for determining rate constants for second order reactions of the form U + V --> W using chemometrics and hard modelling to analyse UV absorption spectroscopic data, where all species absorb with comparable concentrations and extinctions. An interesting feature of this type of reaction is that the number of steps in the reaction is less than the number of absorbing species, resulting in a rank-deficient response matrix. This can cause problems when using some of the methods described in the literature. The approaches discussed in the paper depend, in part, on what knowledge is available about the system, including the spectra of the reactants and product, the initial concentrations and the exact kinetics. Sometimes some of this information may not be available or may be hard to estimate. Five groups of methods are discussed, namely use of multiple linear regression to obtain concentration profiles and fit kinetics information, rank augmentation using multiple batch runs, difference spectra based approaches, mixed spectral approaches which treat the reaction as two independent pseudospecies, and principal components regression. Two datasets are simulated, one where the spectra are quite different and the other where the spectrum of one reactant and the product share a high degree of overlap. Three sources of error are considered, namely sampling error, instrumental noise and errors in initial concentrations. The relative merits of each method are discussed. PMID- 12081046 TI - Fluorimetric analysis of lipase hydrolysis of intermediate- and long-chain glycerides. AB - For the purpose of deducing the digestive behavior of dietary fat in the digestive organs, a fluorimetric method for the measurement of hydrolysis by porcine pancreatic lipase was performed using intermediate- and long- acyl chain glycerides as substrates. Insoluble glycerides constituted by C10-C16 acyl chains were mechanically dispersed in 100% buffer and hydrolyzed by porcine pancreatic lipase. After the reaction, fatty acid released by the enzyme was extracted and its carboxyl group was fluorescently labeled with 9-bromomethylacridine. The 9 acridinylmethyl derivative of the fatty acid was separated and determined by HPLC. The sensitivity of this method was about 1000 times higher than that of the titrimetric method. Only 0.5 ng of porcine pancreatic lipase was sufficient for one routine assay. This assay method was successfully applied to investigate the enzymatic properties of porcine pancreatic lipase with respect to dietary lipids. The effects of some physiological factors concerned with lipid digestion, such as bile salt and colipase, on the lipase hydrolysis were also examined. The method established in the present study could contribute to a highly sensitive assay of some hydrolases containing lipase with regard to insoluble substrates. PMID- 12081047 TI - Sample collection, filtration and preservation protocols for the determination of 'total dissolved' mercury in waters. AB - The objective of the work carried out was to recommend protocols for the collection, filtration (0.45 microm) and preservation of surface water samples for the subsequent determination of total 'dissolved' Hg. Cold vapour (CV) ICP-MS was employed to determine Hg; samples were acidified to a strength of 4 mol l(-1) HCl and 1% NaBH4 was used as the reducing agent in-line. Four types of 125 ml bottles were studied (Teflon, fluorinated ethene propene copolymer, FEP; high density polyethylene, HDPE; polyethylene terephthalate copolyester, PET; polypropylene, PP), together with three cleaning methods (EPA Methods 1631, 1638 and a rinse with reverse osmosis deionised water, 'MilliQ'). The transmission properties of the four materials were also studied to evaluate the potential for contamination from atmospheric Hg0. Results of this bottle study (n = 195), all below the detection limit of 0.5 ng l(-1), indicate that the bottles of choice, from an economic and time-saving perspective, are HDPE and PP, the latter being preferable if the sample is to be stored in a contaminated atmosphere. The bottles would be used on a once-only basis, negating the need for labourious and costly cleaning on repeat use. A simple rinse with MilliQ water would suffice prior to use. Twelve 0.45 microm filter systems (mostly Millipore and Gelman) were studied for (a) their potential Hg contamination properties and (b) their retention of Hg, possibly in colloidal form, during filtration. Ottawa River water, spiked at 50 ng l(-1) Hg, was used as a control sample. Again blank values were all negative, indicating contamination was not a concern but different recoveries of Hg were obtained across the different systems. The optimum systems to use, in that they provided maximum recovery (ca 80%) of Hg, are the Millipore Sterivex capsule and the Millipore Millex disc, both based on the hydrophilic Durapore membrane. The lowest recoveries (23-36%) were found with the Gelman AquaPrep systems and the Millipore HN nylon filter. Four control samples, Ottawa, Rideau and Gatineau Rivers, and a MilliQ blank, were used to study three preservation approaches, in the media: 0.5% BrCl, 2% HCl and 0.04% K2Cr2O7 in 1% HNO3. Mercury was stable for 28 days in a medium of 0.5% BrCl in all four samples but the other two media showed a loss of up to ca 20% Hg over this time frame, the loss being sample and time dependent. This may be a species transformation which is not detected by CV-ICP-MS from an HCl medium but this requires further investigation. Mercury was stable over the 28 days in the spiked blank MilliQ sample for all three preservation media. PMID- 12081048 TI - Comparison of single extraction procedures, using either conventional shaking or microwave heating, and the Tessier sequential extraction method for the fractionation of heavy metals from environmental samples. AB - The conventional four-step sequential extraction method and the EDTA and acetic acid single extraction procedures were applied to sewage sludge and sediment samples. The results obtained with these samples for Cu, Cr, Ni, Pb and Zn using the Tessier method were compared with those supplied by the two single extraction procedures employed. In addition, the Tessier method was also applied to a reference material, CRM 483, and these results were also compared with the certified EDTA and acetic acid values for this sample. As a result, good agreement was found between the metal contents released in the first three fractions of the Tessier method and those leached by the simpler single extraction procedures for the most of the elements studied. Subsequently, the conventional EDTA and acetic acid extraction methods were accelerated by means of microwave energy, in order to reduce the operating time. The extraction efficiency of the first three fractions of the Tessier method was compared with that obtained using the optimised microwave single extraction procedures and only in sewage sludge and CRM 483 samples were satisfactory results found for all the elements studied, except Cr and Pb. This means that the microwave single extraction procedures optimised in this work could be employed as screening methods to evaluate rapidly the easiest mobilizable heavy metals in these samples, although more samples should be analysed to determine their general applicability. The application of the accelerated single extraction procedures to a reference material, CRM 483, provided satisfactory results for all the elements studied, except for Cr in both methods and for Pb in the acetic acid extracts. PMID- 12081049 TI - Collaborative trials of the sampling of two foodstuffs, wheat and green coffee. AB - Two foodstuffs, wheat and green coffee, have been sampled from bulk lots by multiple samplers in the sampling analogue of a collaborative trial. For wheat the variation between samplers, contributing to the standard deviation of sampling reproducibility, was significant for two analytes. No significant sampling reproducibility variation was found in the coffee results, although significant sampling repeatability variation was detected. PMID- 12081050 TI - Editorial--Expectations for the quality and originality of articles submitted to The Analyst. PMID- 12081051 TI - Quality of life. PMID- 12081052 TI - The effect of complete decongestive therapy on the quality of life of patients with peripheral lymphedema. AB - Lymphedema is a chronic disorder which can adversely affect quality of life (QOL). The purpose of this study was 1) to evaluate whether QOL was improved in patients with lymphedema following Complete Decongestive Therapy (CDT), and 2) whether limb volume change as a result of treatment correlated with change in QOL. Thirty-six patients with peripheral lymphedema from varying causes were enrolled in the study. The QOL of each participant, with regard to physical, functional, and psychosocial concerns, was measured by pre- and post-treatment questionnaires. Percent edema volume reduction was calculated for each patient with only one affected limb. QOL pre- and post-treatment scores were assessed by multivariate repeated measures analysis. QOL scores differed significantly (p<0.05) between pre- and posttreatment in all areas of inquiry. Patients with lower extremity lymphedema had significantly greater mean improvement in QOL scores compared with patients with upper extremity lymphedema (p=0.02). There was no correlation between percent edema volume reduction and post-treatment QOL improvement. This study suggests that significant improvements are made in the QOL of patients exhibiting peripheral lymphedema following CDT, which is not necessarily correlated with limb volume reduction. PMID- 12081053 TI - Factors associated with the development of arm lymphedema following breast cancer treatment: a match pair case-control study. AB - We examined factors that may influence the development of arm lymphedema following breast cancer treatment including the specific mode of therapy, patient occupation and life style. Medical record data and a questionnaire were used to collect information after surgery concerning such issues as wound seroma, infection, adjuvant treatment, vessel string (phlebitis), body mass index, smoking habits and stress. Occupational workload was assessed after surgery whereas housework, exercise, hobbies and body weight were assessed both before and after surgery. Seventy-one breast cancer treated women with arm lymphedema lasting more than 6 months but less than 2 years were matched to women similarly treated for breast cancer but without arm lymphedema (controls). The matching factors included axillary node status, time after axillary dissection, and age. In the lymphedema group, there was a higher body mass index at time of surgery (p=0.03) as well at time of study (p=0.04). No differences were found in occupational workload (n=38) or housework, but the lymphedema group reduced their spare time activities including exercise after surgery compared with the controls (p<0.01). In conclusion, women treated for breast cancer with axillary node dissection with or without adjuvant radiotherapy could maintain their level of physical activity and occupational workload after treatment without an added risk of developing arm lymphedema. On the other hand, a higher BMI before and after operation increases the lymphedema risk. PMID- 12081054 TI - Quality of life and peripheral lymphedema. AB - Evaluation of the health-related quality of life (QOL) is becoming commonplace, seeking to provide information about the everyday well-being of a patient. This work examined the QOL of 23 consecutive patients with lymphedema of the upper or lower extremities. Five were men and the other eighteen were women. Their ages ranged from 19 to 74 years (mean 48.3). After clinical examination, and with informed consent of each patient, they were assessed by a psychologist and submitted to a QOL test (SF-36). For controls, twenty-three women and five men with age range of 28 to 66 years (mean 47) were similarly evaluated. The results showed the lymphedema group had a statistically significantly reduced QOL in both physical and mental health as well as social interaction. PMID- 12081055 TI - Changes in mesenteric lymph node T cell phenotype and B and T cell homing properties after murine AIDS infection. AB - We studied MLN (mesenteric lymph nodes) T cell phenotype and MLN T and B cells homing properties after murine AIDS (acquired immunodeficiency syndrome) infection. Our results showed an increase in the percentage of CD4+ cells expressing CD44, CD54 and LPAM-1. There was also a decrease in the proportion of CD8+ cells but an increase in the percentage of CD8+ CD54+ cells. An increased proportion of CD11b+ (Mac1) cells suggested the recruitment of macrophages. Murine AIDS MLN cells labeled with 125I-UDR migrated back to the MLN but did not preferentially migrate to the ILP (intestinal lamina propria). Simultaneous staining for BRDU and IgA confirmed the inability of murine AIDS MLN cells to home to the ILP. These data indicate that murine AIDS infection altered the mucosal immune system while modifying MLN T cell phenotype and MLN T and B cells migratory properties. PMID- 12081056 TI - Treatment of peripheral lymphedema by concomitant application of magnetic fields, vibration and hyperthermia: a preliminary report. AB - Although treatment of peripheral lymphedema has included magnetic fields, vibration, and hyperthermia individually, no one has administered all three at the same time. Accordingly, ten patients with unilateral leg secondary lymphedema were treated using daily therapy for 20 days with each modality for a duration of 60 minutes for a total of 3 hours. The clinical response was evaluated by the coefficient rate of contraction, i.e., by measuring the circumference of the leg 10 cm above and below the patella edge before and after therapy. This preliminary uncontrolled trial resulted in 8 highly effective, 1 improved, and 1 unaffected outcome. In 6 of the 8 highly effective outcomes, the volume of the treated leg became nearly the same as that of the untreated, non-edematous contralateral leg. One patient developed skin erythema which subsided spontaneously several hours after therapy ended. This combined modality therapy for treatment of lymphedema should be explored further. PMID- 12081057 TI - Preliminary evaluation of a new, more simplified physiotherapy technique for lymphatic drainage. AB - The aim of the present study was to evaluate a new, more simplified physiotherapy technique for management of peripheral lymphedema. Fifteen patients (11 female, 4 male) with ages varying from 22 to 63 years (average 54 years) were included in this study. All presented with stage I or II lower limb lymphedema confirmed by lymphoscintigraphy. Each patient underwent a newly designed modified manual lymph drainage technique regularly performed five times a week, one hour per session after one month. The new technique consisted of utilizing sticks, rollers or other cylindrical, flexible, and malleable material which served as a lymph promoting drainage device. All patients demonstrated a highly significant uniform reduction in girth of the leg affected by lymphedema (circumference reduction 2 to 4 cm; average 2.4 cm). After treatment, repeat lymphoscintigraphy suggested a generalized improvement in the scintiscans compared with the initial patterns. This new manual lymph drainage technique efficiently reduces peripheral lymphedema and is less cumbersome and time consuming than standard combined ("decongestive") physiotherapy methods. PMID- 12081058 TI - Lymph-venous shunt in the management of chylous disease (Tosatti-Cariati intervention). PMID- 12081059 TI - Predictors of mortality in adult trauma patients: the physiologic trauma score is equivalent to the Trauma and Injury Severity Score. AB - BACKGROUND: Several statistical models (Trauma and Injury Severity Score [TRISS], New Injury Severity Score [NISS], and the International Classification of Disease, Ninth Revision-based Injury Severity Score [ICISS]) have been developed over the recent decades in an attempt to accurately predict outcomes in trauma patients. The anatomic portion of these models makes them difficult to use when performing a rapid initial trauma assessment. We sought to determine if a Physiologic Trauma Score, using the systemic inflammatory response syndrome (SIRS) score in combination with other commonly used indices, could accurately predict mortality in trauma. STUDY DESIGN: Prospective data were analyzed in 9,539 trauma patients evaluated at a Level I Trauma Center over a 30-month period (January 1997 to July 1999). A SIRS score (1 to 4) was calculated on admission (1 point for each: temperature > 38 degrees C or < 36 degrees C, heart rate > 90 beats per minute, respiratory rate > 20 breaths per minute, neutrophil count > 12,000 or < 4,000. SIRS score, Injury Severity Score (ISS), Revised Trauma Score (RTS), TRISS, Glasgow Coma Score, age, gender, and race were used in logistic regression models to predict trauma patients' risk of death. The area under the receiver-operating characteristic curves of sensitivity versus 1-specificity was used to assess the predictive ability of the models. RESULTS: The study cohort of 9,539 trauma patients (of which 7,602 patients had complete data for trauma score calculations) had a mean ISS of 9 +/- 9 (SD) and mean age of 37 +/- 17 years. SIRS (SIRS score > or = 2) was present in 2,165 of 7,602 patients (28.5%). In single-variable models, TRISS and ISS were most predictive of outcomes. A multiple-variable model, Physiologic Trauma Score combining SIRS score with Glasgow Coma Score and age (Hosmer-Lemenshow chi-square = 4.74) was similar to TRISS and superior to ISS in predicting mortality. The addition of ISS to this model did not significantly improve its predictive ability. CONCLUSIONS: A new statistical model (Physiologic Trauma Score), including only physiologic variables (admission SIRS score combined with Glasgow Coma Score and age) and easily calculated at the patient bedside, accurately predicts mortality in trauma patients. The predictive ability of this model is comparable to other complex models that use both anatomic and physiologic data (TRISS, ISS, and ICISS). PMID- 12081060 TI - Perioperative risk factors for posterior ischemic optic neuropathy. AB - BACKGROUND: Infarction of the optic nerve posterior to the lamina cribrosa, called posterior ischemic optic neuropathy (PION), is a condition that can result in profound bilateral blindness. Cases of PION treated at this institution and those described in the literature were analyzed to identify clinical features that profile those individuals at risk of PION in an attempt to identify major contributing factors that could be addressed prophylactically to enable effective prevention. STUDY DESIGN: Salient clinical features in seven cases of PION diagnosed at the Doheny Eye Institute between 1989 and 1998 are compared with 46 cases of PION reported in the literature. RESULTS: In the Doheny series there were six men and one woman aged 12 to 66 years (mean, 47 years). Five patients were status-post spine surgery, one was status-post knee surgery, and one had a bleeding stomach ulcer. Vision loss was simultaneously bilateral in six of seven patients (85.7%) and was apparent immediately after surgery. There were no abnormal retinal or choroidal findings including diabetic retinopathy, in any of the patients. Notable contributing factors were blood loss in all seven patients, ranging from 2,000 to 16,000 mL, with a drop in hematocrit of 9.5% to 19% (mean, 14%), and intraoperative systemic hypotension in all patients. Facial edema was a factor in three of six spine surgery patients (50%). Patients reported in the literature had a mean age of 50 years and were also predominantly men (34 of 46, 74%) who underwent spine surgery (30 of 46, 65.2%). CONCLUSIONS: Middle-aged men undergoing spine surgery with prolonged intraoperative hypotension and postoperative anemia and facial swelling are at risk of developing PION from hypovolemic hypotension. Avoiding or immediately correcting these contributory factors can reduce the incidence of PION. PMID- 12081061 TI - Well-differentiated thyroid carcinoma in Hong Kong Chinese patients under 21 years of age: a 35-year experience. AB - BACKGROUND: Sporadic well-differentiated thyroid cancer is an uncommon condition in children, adolescents, and young adults. It is associated with distinct clinicopathologic features and prognosis. The present study reviews our experience in management of this condition in ethnic Chinese in Hong Kong. STUDY DESIGN: A retrospective study was performed to review the clinicopathologic features and outcomes of 34 patients less than 21 years of age with well differentiated thyroid carcinoma over a 35-year period. Median followup was 15.2 years (range 1 to 32.5 years). RESULTS: There were 27 girls and 7 boys with a median age of 19 years (range 10 to 21 years). None had previous history of irradiation. Twenty-eight patients had papillary and six had follicular carcinomas. Operative procedures included total thyroidectomy (n = 27) and unilateral lobectomy (n = 7) with concomitant neck dissection performed in nine patients. The median tumor size was 2 cm and extrathyroidal invasion was present in 19 tumors. Adjuvant radioactive iodine treatment was administered to 18 patients after total thyroidectomy. Disease progression or recurrence rate was 24% and 27% at 5 and 10 years, respectively. The presence of lymph node metastases was associated with a higher incidence of disease recurrence. One patient with advanced local disease died from tumor bed recurrence with anaplastic transformation 18 years after the initial operation. CONCLUSIONS: Well differentiated thyroid carcinoma is a relatively indolent tumor associated with good prognosis in young patients. Although death from this condition is rare, recurrence is frequent and longterm followup is necessary. PMID- 12081062 TI - Bleeding esophagogastric varices from extrahepatic portal hypertension: 40 years' experience with portal-systemic shunt. AB - BACKGROUND: This article discusses the largest and longest experience reported to date of the use of portal-systemic shunt (PSS) to treat recurrent bleeding from esophagogastric varices caused by extrahepatic portal hypertension associated with portal vein thrombosis (PVT). STUDY DESIGN: Two hundred consecutive children and adults with extrahepatic portal hypertension caused by PVT who were referred between 1958 and 1998 after recovering from at least two episodes of bleeding esophagogastric varices requiring blood transfusions were managed according to a well-defined and uniformly applied protocol. All but 14 of the 200 patients were eligible for and received 5 or more years of regular followup (93%); 166 were eligible for and received 10 or more years of regular followup (83%). RESULTS: The etiology of PVT was unknown in 65% of patients. Identifiable causes of PVT were neonatal omphalitis in 30 patients (15%), umbilical vein catheterization in 14 patients (7%), and peritonitis in 14 patients (7%). The mean number of bleeding episodes before PSS was 5.4 (range 2 to 18). Liver biopsies showed normal morphology in all patients. The site of PVT was the portal vein alone in 134 patients (76%), the portal vein and adjacent superior mesenteric vein in 10 patients (5%), and the portal and splenic veins in 56 patients (28%). Postoperative survival to leave the hospital was 100%. Actuarial 5-year, 10-year, and 15-year survival rates were 99%, 97%, and 95%, respectively. Five patients (2.5%), all with central end-to-side splenorenal shunts, developed thrombosis of the PSS, and these were the only patients who had recurrent variceal bleeding. During 10 or more years of followup, 97% of the eligible patients were shown to have a patent shunt and were free of bleeding. No patient developed portal systemic encephalopathy, liver function tests remained normal, liver biopsies in 100 patients showed normal architecture, hypersplenism was corrected. CONCLUSION: PSS is the only consistently effective therapy for bleeding esophagogastric varices from PVT and extrahepatic portal hypertension, resulting in many years of survival, freedom from recurrent bleeding, normal liver function, and no encephalopathy. PMID- 12081064 TI - Early debridement for necrotizing pancreatitis: is it worthwhile? AB - BACKGROUND: The timing for debridement of necrotizing pancreatitis is controversial. We reviewed our experience with early and delayed surgical debridement in patients with necrotizing pancreatitis. STUDY DESIGN: The records of patients diagnosed with acute necrotizing pancreatitis from January 1993 through June 2000 were reviewed retrospectively. Data were analyzed with respect to Ranson's, APACHE II, and multiple organ failure scores, etiology, presence of infection, overall and ICU length of stay, time to first debridement, number of debridements, fluid requirements, days to enteral feeding, transfusion requirements, complications, and mortality. RESULTS: Twenty-six patients (18 males, 8 females, mean age 51 years) were diagnosed with acute necrotizing pancreatitis. The admission Ranson's score was 4.8, the APACHE II score was 11.7, and multiple organ failure score was 4.2. All but one patient underwent pancreatic debridement (4.3 debridements per patient). Eighteen patients (69%) had infected pancreatic necrosis. The timing of debridement was based on patients' condition and surgeon's preference. The presentation and demographics of patients who underwent early (<2 weeks) or late (>2 weeks) debridement did not differ significantly. Patients debrided early had a trend toward higher mortality (29% versus 18%) and experienced a higher number of major complications (p < 0.05). The six patients (23%) who died were older, had multiple organ failure scores, and more often had Candida in the infected necrosis (p < .05). CONCLUSIONS: Early debridement for acute necrotizing pancreatitis might not improve survival and might even be associated with increased number of complications. Most patients diagnosed with necrotizing pancreatitis eventually need debridement, but it might be beneficial to delay debridement if the patient's condition allows for it. PMID- 12081063 TI - Endotoxin potentiates hepatocyte apoptosis in cholestasis. AB - BACKGROUND: Cholestasis is a component of liver disease of almost any etiology, including septic liver injury. The cellular mechanisms of liver injury in cholestasis and sepsis remain unresolved. We evaluated apoptosis, a well orchestrated and potentially reversible mechanism of cell death, in bile duct ligated and endotoxin-injected rats. STUDY DESIGN: Male Sprague-Dawley rats were randomly assigned to six groups (n = 6-9): bile duct-ligated+endotoxin (B+E), sham+endotoxin (S+E), bile duct-ligated (B), sham (S), endotoxin (E), and normal (N). On day 1, the bile ducts of B+E and B rats were ligated and severed. S+E and S animals underwent biliary manipulation only. On day 3, B+E, S+E, and E groups received 3 mg/kg endotoxin i.v.. On day 4, livers from all rats were excised, fixed, and stained (hematoxylin and eosin and terminal deoxynucleotidyl transferase dUTP nick-end labeling [TUNEL]). Portions were frozen for DNA fragmentation analysis. Caspase 3 activity was determined using isolated hepatocytes. RESULTS: Livers from all groups (B+E, S+E, E, and B) except normal and sham displayed apoptosis by hematoxylin and eosin staining, TUNEL staining, and DNA fragmentation. Histologic evaluation revealed 10% to 20% necrosis in endotoxin-treated animals (B+E, S+E, and E). Caspase 3 activity was significantly higher in endotoxin-treated animals versus animals without endotoxin (treated 0.450 +/- 0.08 versus nontreated 0.135 +/- 0.05, p < 0.0001) (mean +/- SD). CONCLUSIONS: Cholestatic livers had apoptosis without progression to necrosis. When exposed to the second insult of endotoxin, cholestatic livers received an acute on chronic apoptotic trigger, and proceeded to necrosis. Endotoxin was a potent hepatotoxic insult because all treated rat livers displayed both apoptosis and necrosis. PMID- 12081065 TI - Prospective trial of a blood supply-based technique of pancreaticojejunostomy: effect on anastomotic failure in the Whipple procedure. AB - BACKGROUND: Anastomotic failure at the pancreaticojejunostomy after a Whipple procedure, manifested either as a pancreatic fistula or intraabdominal abscess, is still an unacceptably common postoperative complication. STUDY DESIGN: A prospectively collected series of 123 patients underwent a Whipple procedure. During the pancreaticojejunostomy, the blood supply at the cut surface of the pancreas was evaluated, and if deemed inadequate, the pancreas was cut back 1.5 to 2.0 cm to improve the blood supply. The anastomosis was performed under magnification with meticulous technique. RESULTS: There were 123 Whipple procedures performed. In 47 (38%), the blood supply was considered inadequate and the pancreas was cut back. Postoperatively, there were 2 pancreatic fistulas (1.6%) and 2 intraabdominal abscesses (1.6%). There was 1 (0.8%) postoperative death from aspiration pneumonia. CONCLUSIONS: Pancreatic fistula, the most serious complication of the Whipple procedure, can be almost entirely eliminated by a technique that combines meticulous attention to placement and tying of sutures under magnification with optimization of blood supply to the anastomosis. PMID- 12081066 TI - Blind distal pancreatectomy for occult insulinoma, an inadvisable procedure. AB - BACKGROUND: Fasting hypoglycemia with neuroglycopenic symptoms corrected by administration of glucose are the hallmarks for the diagnosis of insulinoma. Surgical resection is the treatment of choice for insulinomas, but localization of these lesions can be challenging. Blind distal pancreatectomy has been advocated for occult insulinomas not detected on imaging studies or during abdominal exploration. With the advent of newer localization techniques, we challenge the wisdom of this approach. STUDY DESIGN: The records of patients (multiple endocrine neoplasia excluded) with pathologically proved insulinoma who were screened at our institution or referred to us after a failed blind distal pancreatectomy were reviewed. All records included patient history and results of physical examination and routine blood and urine tests. The diagnosis of insulinoma was confirmed during a supervised fast. Patients with biochemically proved insulinoma underwent localization studies and operation. Studies included CT scans, MRI, transabdominal ultrasound, intraoperative ultrasonography, angiography (more recently, Ca++-stimulated arteriography), and venous sampling. RESULTS: From 1970 to 2000, 99 patients (34 men, 65 women; mean age 43 years) underwent operation. All patients with benign tumors (92) were cured after operation. Seventeen patients were referred to the NIH after a failed blind distal pancreatectomy. Of these, 5 were diagnosed as having factitious hypoglycemia. In the other 12 patients a tumor was localized in the pancreatic head. Two patients incorrectly diagnosed with nesidioblastosis after initial surgery were subsequently cured by resection of an insulinoma. CONCLUSIONS: The use of preoperative imaging studies, most notably Ca++-stimulated arteriography, and intraoperative ultrasonography permits detection of virtually all insulinomas, including reopcrated cases. When a tumor is not detected, the procedure should be terminated and the patient referred to a center capable of performing advanced preoperative and intraoperative localization techniques. With the preoperative and intraoperative imaging strategies currently available, the use of blind distal pancreatectomy for occult insulinoma should be abolished. PMID- 12081067 TI - Non-anti-Gal alpha1-3Gal antibody mechanisms are sufficient to cause hyperacute lung dysfunction in pulmonary xenotransplantation. AB - BACKGROUND: Hyperacute lung dysfunction, which is always associated with pulmonary pig-to-primate xenotransplantation is not well understood. The mechanisms associated with its occurrence seem to differ from mechanisms involved in hyperacute xenograft rejection seen in porcine hearts or kidneys transplanted into primates. To determine the contribution of anti-Gal alpha1-3Gal antibodies (alphaGAb) in such a process, we performed a set of orthotopic pig lung transplants into baboons depleted of alphaGAb and compared graft function and survival with those receiving only immunosuppression. STUDY DESIGN: Pigs expressing human membrane cofactor protein served as donors. All baboons received triple immunosuppressive therapy. Depletion of alphaGAb in the experimental group (n = 4) was done by way of immunoadsorption using immunoaffinity membranes. Controls (n = 4) did not undergo immunoadsorption. Orthotopic lung transplants were performed through a left thoracotomy. Main pulmonary artery blood flow and pressure, left pulmonary artery blood flow, and left atrial pressure were recorded. RESULTS: At 1 hour after reperfusion, pulmonary artery graft flows and pulmonary vascular resistances (PVR) were better in animals depleted of alphaGAb than in controls (605 +/- 325.2 mL/min versus 230 +/- 21 mL/min; 27.1 +/- 41.3 mmHg/L/min versus 63 +/- 1 mmHg/L/min). But at 3 hours after reperfusion average graft flows in baboons depleted of alphaGAb had decreased to 277.6 +/- 302.2 mL/min and PVRs had increased 58.3 +/- 42.0 mmHg/L/min. On the other hand, controls maintained stable flows and PVRs (223 +/- 23 mL/min; 61 +/- 3 mmHg/L/min). Survival was ultimately better in control baboons when compared with alphaGAb depleted ones (12.2 +/- 3.3 h versus 4.4 +/- 3.2 h). CONCLUSION: Unlike heart and kidney xenograft transplants, hyperacute lung xenograft dysfunction seems to be mediated by factors other than alphaGAb. PMID- 12081068 TI - Middle aortic syndrome: distal thoracic and abdominal coarctation, a disorder with multiple etiologies. AB - BACKGROUND: Congenital coarctation of the thoracic aorta at the ligamentum arteriosum or the aortic arch is well recognized. But a much less common variety (0.5% to 2.0%) of aortic coarctation is located in the distal thoracic aorta, or abdominal aorta, or both and is often called "middle aortic syndrome" or "mid aortic dysplastic syndrome." These types of aortic coarctation are most often secondary to a form of granulomatis vasculitis commonly known as Takayasu's disease in this country or aortitis syndrome in Japan. No single genesis explains every case and beside vasculitis as a cause, some are thought to be congenital in origin and others are associated with von Recklinghausen's disease. STUDY DESIGN: Eight patients with severe hypertension or claudication secondary to middle aortic coarctation were studied with aortograms and subsequently treated by vascular reconstruction procedures. RESULTS: Vascular reconstructions consisted of aortoaortic bypass, aortic resection with interposed grafting, reanastomotic resection of renal arteries into prosthetic grafts, and renal artery bypass with autogenous material. All eight patients' grafts have remained patent, with followups of 4 to 9 years, with relief of hypertension and claudication. Although Takayasu's disease can be progressive, aggressive surgical treatment in eight patients followed for 4 to 8 years postoperatively demonstrates that severe hypertension, claudication, or both are important indications for revascularization. CONCLUSIONS: Whatever the cause, assuming that active aortic inflammation has been medically treated and is in a burned-out state, patients with abdominal coarctation who have symptomatic renovascular hypertension, claudication, or both are good candidates for revascularization. Although surgical repair is more difficult than with congenital thoracic coarctation, because aortic walls are fibrotic and often also involve the renals, all eight of our patients had successful longterm correction of their hypertension and coarctation. PMID- 12081069 TI - What's new in neurological surgery. PMID- 12081070 TI - What's new in orthopaedic surgery. PMID- 12081071 TI - Ethical regulations for innovative surgery: the last frontier? AB - BACKGROUND: There are no clear federal regulations governing innovative surgery, even though general guidelines regulating research with human subjects do exist. We hypothesized that US surgeons are unaware of Department of Health and Human Services regulations, rarely seek IRB review, generally oppose outside regulation of innovative surgery, and are uncertain what constitutes innovation and research. These circumstances, if true, would pose a significant ethical problem and present potential harm to patients as unwitting subjects of research. STUDY DESIGN: In a pilot study we reviewed 527 issues of US surgical and medical journals, selecting 59 articles published between 1992 and 2000, that described innovative surgery. Corresponding authors from university hospitals (71%) and other facilities (29%) were sent an anonymous questionnaire. RESULTS: The survey was conducted between November 2000 and May 2001. Twenty-one questionnaires were returned, completed with responses, constituting a 35% overall response rate. Fourteen authors confirmed their work was research, yet only six had sought prior IRB review. The majority of authors (15 of 21) did not submit their protocol to IRB. Only seven authors had mentioned the innovative nature of the procedure in the informed consent form. Seven authors claimed familiarity with Office for Human Research Protections definitions of research and human subject. Two-thirds of the respondents stated that government regulations for the protection of human subjects of innovative surgery would not be appropriate. CONCLUSIONS: The current system of definitions, ethical theories, and voluntary professional guidelines may be inadequate to meet the challenge of surgical innovation. Further research is proposed to examine the adequacy of the existing guidelines. PMID- 12081072 TI - A review of surgical care in space. PMID- 12081073 TI - Radiofrequency ablation of unresectable primary liver cancer. PMID- 12081074 TI - Mammillary fistula as a manifestation of acne inversa (hidradenitis suppurativa): report of two cases. PMID- 12081075 TI - The paradox of hydration in advanced terminal illness. PMID- 12081076 TI - Primary melanoma of the esophagus. PMID- 12081077 TI - Traumatic carotid cavernous fistula. PMID- 12081079 TI - Lymphoscintigraphic anatomy of sentinel lymphatic channels. PMID- 12081078 TI - Ex vivo right trisegmentectomy with reconstruction of inferior vena cava and "flop" reimplantation. PMID- 12081080 TI - Scientific data from clinical trials: investigators' responsibilities and rights. PMID- 12081081 TI - Corporal punishment by parents and associated child behaviors and experiences: a meta-analytic and theoretical review. AB - Although the merits of parents using corporal punishment to discipline children have been argued for decades, a thorough understanding of whether and how corporal punishment affects children has not been reached. Toward this end, the author first presents the results of meta-analyses of the association between parental corporal punishment and 11 child behaviors and experiences. Parental corporal punishment was associated with all child constructs, including higher levels of immediate compliance and aggression and lower levels of moral internalization and mental health. The author then presents a process-context model to explain how parental corporal punishment might cause particular child outcomes and considers alternative explanations. The article concludes by identifying 7 major remaining issues for future research. PMID- 12081082 TI - Ordinary physical punishment: is it harmful? Comment on Gershoff (2002). AB - E. T. Gershoff (2002) reviewed processes that might mediate and contexts that might moderate the associations between corporal punishment (CP) and child behaviors and provided an account of the methodological weaknesses of the research reviewed in her meta-analyses. In this examination of Gershoff, the authors argue that the biases and confounds in the meta-analyses further limit any causal inferences that can be drawn concerning the detrimental "effects" of CP on associated child behaviors. The authors suggest that undesirable child outcomes are associated with CP because the construct marks inept harsh parenting and conclude that although the harmful effects of physical abuse and other extreme punishments are clear, a blanket injunction against spanking is not justified by the evidence presented by Gershoff. PMID- 12081083 TI - Perspectives on the effects of corporal punishment: comment on Gershoff (2002). AB - Parental use of corporal punishment has been a contentious child-rearing topic for thousands of years. E. T. Gershoff's (2002) meta-analyses undoubtedly will rekindle the debate over whether parents should spank their children. The quality of the existing data is discussed as well as several surprising findings from the meta-analyses. The effects of corporal punishment are then considered from the perspectives of learning theory, socialization theory, and the child's point of view. Her ecological-process model is then evaluated in light of these perspectives. Research directions and social policy implications are addressed. PMID- 12081084 TI - Punishment revisited--science, values, and the right question: comment on Gershoff (2002). AB - In this comment on E. T. Gershoff (2002), the author notes the historical phases of punishment research. Punishment as a disciplinary tactic is best viewed as a packaged variable and therefore needs to be investigated in the context of other socialization practices. The role of parental values in this debate about punishment utilization and effectiveness merits more consideration. New directions in punishment research are also noted. These include the need for a family-systems perspective, a family-typology approach, a transactional model of punishment, the use of innovation observation and self-report methods, and more culturally sensitive paradigms. PMID- 12081086 TI - Self-focused attention and negative affect: a meta-analysis. AB - This meta-analysis synthesized 226 effect sizes reflecting the relation between self-focused attention and negative affect (depression, anxiety, negative mood). The results demonstrate the multifaceted nature of self-focused attention and elucidate major controversies in the field. Overall, self-focus was associated with negative affect. Several moderators qualified this relationship. Self-focus and negative affect were more strongly related in clinical and female-dominated samples. Rumination yielded stronger effect sizes than nonruminative self-focus. Self-focus on positive self-aspects and following a positive event were related to lower negative affect. Most important, an interaction between foci of self attention and form of negative affect was found: Private self-focus was more strongly associated with depression and generalized anxiety, whereas public self focus was more strongly associated with social anxiety. PMID- 12081085 TI - When and where do we apply what we learn? A taxonomy for far transfer. AB - Despite a century's worth of research, arguments surrounding the question of whether far transfer occurs have made little progress toward resolution. The authors argue the reason for this confusion is a failure to specify various dimensions along which transfer can occur, resulting in comparisons of "apples and oranges." They provide a framework that describes 9 relevant dimensions and show that the literature can productively be classified along these dimensions, with each study situated at the intersection of various dimensions. Estimation of a single effect size for far transfer is misguided in view of this complexity. The past 100 years of research shows that evidence for transfer under some conditions is substantial, but critical conditions for many key questions are untested. PMID- 12081087 TI - Preserving the ethical propriety of statistical devices. AB - C. D. Herrera (1996) introduced an innovative argument against the use of deception in psychological research. In essence, Herrera contended that because of the presumed problems with null hypothesis statistical testing, researchers could not justify their continued use of deception in research. Although this is an interesting argument, there are several alternative perspectives that must be considered. In examining these alternatives, the author concluded that psychologists may continue to use deception under certain circumstances outlined in the American Psychological Association's ethical code of conduct. PMID- 12081088 TI - The role of suggestions and personality characteristics in producing illness reports and desires for suing the responsible party. AB - For this project, 92 students entered an abandoned theater room in an old basement of the university where sand was scattered throughout. The purpose of the study was to experimentally demonstrate that psychological suggestions could produce illness reports and to explore who is most likely to say that they would sue for personal damages. The students filled out the Trait-State Anger Scale and two subscales, Anger Temperament and Anger Reaction (C. D. Spielberger, G. A. Jacobs, S. Russell, & R. S. Crance, 1983) as well as the Costello-Comrey Anxiety Scale (G. C. Costello & A. L. Comrey, 1967), the Hardiness Inventory (S. C. Kobasa, 1982), the Pennebaker Inventory of Limbic Languidness (J. W. Pennebaker, 1982), and, embedded in the Hardiness Inventory, measures of current illness as a result of exposure to the basement room. Half the participants were met by a confederate student who claimed to be cleaning up the remains of a production of "Lawrence of Arabia," and the other half were met by a confederate construction worker who claimed that "The stuff will tear up your skin and your lungs." Those in the experimental groups who perceived danger and scored low in the hardiness dimension of challenge were more likely to report symptoms of illness. Willingness to file a law suit was predicted by a model including perceived danger and the personality characteristic of anger reactivity. PMID- 12081089 TI - The accuracy of abstracts in psychology journals. AB - This article provides an empirically supported reminder of the importance of accuracy in scientific communication. The authors identify common types of inaccuracies in research abstracts and offer suggestions to improve abstract article agreement. Abstracts accompanying 13% of a random sample of 400 research articles published in 8 American Psychological Association journals during 1997 and 1998 contained data or claims inconsistent with or missing from the body of the article. Error rates ranged from 8% to 18%, although between-journal differences were not significant. Many errors (63%) were unlikely to cause substantive misinterpretations. Unfortunately, 37% of errors found could be seriously misleading with respect to the data or claims presented in the associated article. Although deficient abstracts may be less common in psychology journals than in major medical journals (R. M. Pitkin, M. A. Branagan, & L. F. Burmeister, 1999), there is still cause for concern and need for improvement. PMID- 12081090 TI - Specificity of measures of thought control. AB - The rebound effect (D. M. Wegner, 1994) essentially involves the persistence of thought when an effort has been made to suppress the cognitive activity. Some of the research in this area has been directed at determining personal factors that predict differential levels of rebound. To this end, measures of thought control have emerged. In the present study the authors examined the properties of 2 measures of thought control: the Thought Control Questionnaire (TCQ; A. Wells & M. I. Davies, 1994) and the White Bear Suppression Inventory (WBSI; D. M. Wegner & S. Zanakos, 1994). Each measure was examined in relation to measures of obsessive-compulsive symptoms and worry, as well as combined factorial structure. Scores generally indicated that items on the Worry and Punishment subscales of the TCQ were highly related to worry and obsessive-compulsive symptoms, whereas scores on the WBSI showed an association between Worry and 1 subscale (Slowness) of obsessive-compulsive symptoms. Results of a principal components analysis indicate that, in the aggregate, the items from the TCQ and WBSI form 2 factors: Dysfunctional and Functional Thought Control Strategies. From these data, it appears that the TCQ possesses greater potential to predict performance in rebound experiments and has greater clinical usefulness. Future research directions involving the TCQ are recommended. PMID- 12081091 TI - Empathy theory and practice: a survey of psychologists and counselors. AB - Psychologists and counselors completed a questionnaire devised by the authors to explore the relationships between their extent of endorsement of empathy definitions, their use and views of empathy, and their identification with theories of psychotherapy. Results from 565 respondents suggested that those who identified with humanistic/experiential and psychodynamic theories seemed to have similar views of how empathy is defined and viewed and reported that they use empathy more than those with other theoretical inclinations. These findings suggest that there is some consistency between theoretical identification and definition, as well as reported use and views of empathy. PMID- 12081092 TI - Perceived attributes of health-promoting laughter: a cross-generational comparison. AB - A small but growing body of empirical data support the popular belief that laughter benefits health. However, there are many varieties of laughter and no reason to assume all varieties should be, or would be, perceived as equally beneficial. The authors examined which types of laughter and which characteristics of laughter people associate with health and whether there are generational differences in this perception. Young adults and older participants rated 12 laughter terms (e.g., chuckle, giggle, belly laugh) on their contribution to health and assessed various aspects of each laughter type on 11 scales. The young adults characterized health-promoting laughter as strong, active, uninhibited, and involving movement; the older participants characterized it as socially appropriate. Both groups associated health-promoting laughter strongly with positive emotion and absence of malice. Implications for placebo effects in laughter interventions are discussed. Differences between humor and laughter and among the proposed mechanisms by which they affect health are clarified. PMID- 12081093 TI - The relationship between emotional intelligence and ego defense mechanisms. AB - The author examined the relationship between the components of emotional intelligence (perception of emotion, affect regulation, and emotional knowledge) and personality factors associated with adaptation, represented by the hierarchical model of defense mechanisms (M. Bond, S. Gardner, J. Christian, & J. Sigal, 1983). Bivariate correlation analyses yielded mixed results; the adaptive defense styles were correlated with overall emotional intelligence but not with the emotional perception and regulation components, as was hypothesized. Emotional knowledge was correlated with both adaptive and maladaptive defense styles and with general intelligence, as was expected. Implications for counseling and psychoeducational interventions are suggested. PMID- 12081094 TI - Leadership styles and occupational stress among college athletic directors: the moderating effect of program goals. AB - The interaction between an individual's abilities and the perceived demands of the workplace appears to make a unique contribution to job-related stress above and beyond that of dispositional or situational factors alone (R. S. Lazarus, 1990). In the present study, the author evaluated this contention among 245 male intercollegiate athletic directors by assessing the combined influence of leadership style and program goals on occupational stress. Regression analyses revealed the presence of both significant main effects and interaction effects of leadership style and program goals in the prediction of emotional exhaustion, daily job stress, and personal accomplishment. Findings are discussed in terms of person-environment fit theory (J. R. P. French, R. D. Caplan, & R. V. Harrison, 1982) and the notion of perceived control within the occupational setting. PMID- 12081095 TI - The relationships among Black consciousness, self-esteem, and academic self efficacy in African American men. AB - The purpose of this study was to investigate the relationships among Black consciousness, self-esteem, and academic self-efficacy in African American men. The participants were 120 African American male college students at a predominantly African American university. The authors administered 3 instruments -the Developmental Inventory of Black Consciousness (DIB-C; J. Milliones, 1980), the M. Rosenberg (1965) Self-Esteem Scale, and R. E. Wood and E. A. Locke's (1987) Academic Self-Efficacy Scale--to test the hypotheses. They used an independent-measures t test and a Pearson r correlation to analyze the data. The results of the study supported the hypotheses under investigation. Significant positive relationships were found between Black consciousness and self-esteem and Black consciousness and academic self-efficacy. The results of the study showed that Black consciousness appears to be an important construct to use in understanding self-esteem and academic self-efficacy in African American men. PMID- 12081096 TI - An introduction to Chinese psychology--its historical roots until the present day. AB - In this article the authors review the historical development of Chinese psychology. China's long history as a country immersed in the study of psychological issues is well known and has had an influence on developments in the field of psychology around the world. Modern Chinese psychology, however, was imported from the West and the Soviet Union and has been closely linked with China's social environment and changes of national policy. Today Chinese psychology is still in a preliminary stage. Despite difficulties that may impede its future development, the importance of psychology in the modernization of China has been widely recognized. PMID- 12081097 TI - HCV and GBV-c/HGV infection in HIV positive patients in southern Italy. AB - Flaviviridae-hepatitis C virus (HCV) and GB virus C/hepatitis G virus (GBV-C/HGV) -and human immunodeficiency virus (HIV) frequently show similar modes of transmission. HCV and GBV-C/HGV infection was assessed in 134 consecutive patients with evidence of HIV infection, living in Campania, Italy. Data obtained from this cohort were compared with those obtained from 252 age- and sex-matched HCV infected patients without evidence of HIV infection (HCV control group). Following enzymatic immunoassays, samples were tested for the presence of HCV-RNA by RT-PCR. The HCV-RNA positive sera were genotyped by LiPA procedure. The prevalence of HCV infection in HIV patients was 19.40% and the largest group of HIV-HCV co-infected patients (84.62%) was represented by intravenous drug users (IVDU). The distribution of HCV genotypes in HIV-HCV patients was different, compared to that observed in HCV control group. HCV genotypes la (50%) and 3a (23.08%) were more frequently detected in HIV HCV patients, compared to HCV control group (5.16 and 5.56% for la and 3a, respectively). Conversely, HCV genotypes lb (55.70%) and 2a/2c (30.26%) were more represented in HCV control group, compared to HIV-HCV patients (15.38 and 0% for lb and 2a/2c, respectively). GBV-C/HGV seroprevalence was 41.04% in HIV patients and 6.54% in healthy control individuals. Differently from HCV, GBV-C/HGV infection did not correlate to a preferential risk behaviour in the HIV cohort. Comparative analysis of HCV and GBV-C/HGV infection indicates that the use of injecting drugs might play a key role in the epidemiology of HCV and, in particular, of la and 3a HCV genotypes, in HIV patients. PMID- 12081098 TI - Does non-responder bias have a significant effect on the results in a postal questionnaire study? AB - BACKGROUND AND AIM: In epidemiological questionnaire studies results can be influenced by non-responder bias. However, in respiratory epidemiology this has been analysed in very few recently published papers. The aim of our paper is to assess if the results found in our previous postal questionnaire study in an adult population in Northern Finland were biased by non-response. METHODS: A random sample of 385 persons from the 1,284 non-responders in a previous postal questionnaire study was examined. The same questionnaire as in the original study was again mailed to these persons, and those still not answering were contacted by phone. RESULTS: Totally 183 complete answers (48%) were collected. Lack of interest (56%) and forgetting to mail the response letter (22%) were the most common reasons to non-response. Typical non-responders were young men and current smokers who less frequently reported respiratory symptoms in exercise and asthma than the responders in the original study. Answers collected by phone gave for some questions higher prevalence rates than postal answers. CONCLUSION: Firstly, in this population the response rate (83.6%) in the original study was high enough to provide reliable results for respiratory symptoms and diseases, only the prevalence of current smoking was biased by non-response. Secondly, the methods used for collecting responses in a non-response study may influence the results. PMID- 12081100 TI - Helicobacter pylori colonization and diarrhoeal illness: results of a population based cross-sectional study in adults. AB - It has been suggested that Helicobacter pylori colonization may protect against diarrhoeagenic gastrointestinal infections. The aim of this analysis was to investigate the association between H. pylori infection and the frequency of diarrhoeal episodes among adults. Helicobacter pylori infection status was determined by 13C-urea breath test. Overall, 784 adults (mean age: 48.7 +/- 17.7; range 18-85 years) who participated in two epidemiological studies were included in the analysis. Overall H. pylori prevalence was 25.5%. Episodes of diarrhoea within prior 3 months were less often reported for H. pylori infected subjects compared with H. pylori negative subjects (40.2 vs. 51.6%, p = 0.016). Compared to H. pylori negative subjects the odds ratio (OR) for the occurrence of diarrhoea within the prior 3 months was 0.63 (95% CI: 0.45-0.87) for H. pylori infected subjects. After adjustment for covariates the OR was 0.67 (95% CI: 0.47 0.95). These results support the hypothesis that colonization with H. pylori may protect from gastrointestinal infections that cause diarrhoea. PMID- 12081099 TI - Decrease in human fascioliasis in Gipuzkoa (Spain). AB - From 1981 to 2000 sixty-one cases of human fascioliasis were detected in the province of Gipuzkoa (Spain), most of them during the period of parenchymal invasion. Four family outbreaks were recorded. Thirty, 15, 13 and three cases were detected during the four consecutive 5-year periods. The decrease in this infection over the years is probably related to the change in eating habits. PMID- 12081101 TI - Indicators of fetal growth and infectious disease in childhood--a birth cohort with hospitalization as outcome. AB - We evaluated the association between indicators of fetal growth and hospitalization with infectious disease during childhood in a cohort of 10,400 newborns. The cohort was based on children born to mothers who at about 36 weeks of gestation attended the midwife centres in Odense and Aalborg, Denmark for a routine examination. Women were recruited to the study from April 1984 to April 1987. After linkage with the National Hospital Registry, the first hospitalization with infectious disease from 6 months up to 12 years of age was identified. The cumulative incidence of hospitalization with infectious disease during follow-up was 18.9%. Preterm birth was associated with an increased risk of being hospitalized with infections during childhood (incidence rate ratio: 1.67, 95% CI: 1.33-2.10); low birth weight had a similar association, but only in preterm birth. Reduced birth length related to the head was correlated with an increased risk of hospitalization with infections. The effect of gestational age was mainly seen in the period close to the time of birth, but the children who were short at birth appeared to remain at increased risk throughout the age interval under analysis. In conclusion, the study suggests that preterm birth was the main factor underlying the association between low birth weight and the increased risk of hospitalization with infectious disease during childhood. However, it could not explain the increased risk in children who were short at birth. PMID- 12081102 TI - Predictive factors of mental disorders in patients with suspected ischaemic cardiopathy. AB - OBJECTIVE: To identify the predictive factors of mental disorders in patients with suspected Ischaemic Cardiopathy (IC). METHOD: The patients admitted to the Cardiology Service of an University Hospital (SW Spain) for acute thoracic pain with suspected IC were studied. Patients were classified into four diagnostic groups (acute myocardial infarction, unstable angina, non-ischaemic cardiological process, non-cardiological) based on clinical, electrocardiographical and biochemical data. The sociodemographic and clinical variables were obtained by a 'ad hoc' questionnaire and the clinical records of the patient. The GHQ-28 questionnaire was used to assess the status of mental health, and a score of > or = 6 was taken as the cut-off point for being a 'probable psychiatric case'. Crude odds ratios (OR +/- 95% CI), and adjusted OR according to a logistic model, were calculated in order to study the variables associated with the mental disorders. The tabulation and analysis of data was carried out with the SPSS v.8 (for Windows) program. RESULTS: From June 1996 to November 1997, 185 patients were studied; 72% were males and the mean age was 60.2 years (SD = 10.4). 49.2% of the patients were scored at > or = 6 on the GHQ-28, and the sub-scale accounting for the highest scores was that of somatic symptoms of psychological origin. The adjusted ORs showed that female sex (OR: 2.5; 95% CI: 1.2-5.0), previous personal history of IC (OR: 2.3; 95% CI: 1.1-4.4), and the presence of arterial hypertension (OR: 2.0; 95% CI: 1.1-3.9), were the predictive variables for being a 'probable psychiatric case', whereas neither age nor comorbidity were predictive. CONCLUSIONS: The percentage of subjects considered to be a 'probable psychiatric case' among patients with suspected ischaemic cardiopathy was high. Screening for mental disorders and intervention to control the risk factors associated are measures recommended in these patients. PMID- 12081103 TI - Premature avoidable deaths by road traffic injuries in Belgium: trends and geographical disparities. AB - Road traffic injuries are a major public health problem. In this study, we chose Years of Potential Life Lost (YPLL) to analyse the trends during the period 1974 1994 and the relative impact of the traffic injuries death on total mortality and on total avoidable mortality in Belgium. We analysed the geographical trends over a 20-year period at the district level. The YPLL age-adjusted rates were analysed for four successive 5-year periods: 1974-1978, 1980-1984, 1985-1989, 1990-1994 and the ratios of YPLL rates were used to describe changes between 1974 and 1994 at district level. The YPLL rates decrease for 'all causes mortality', 'total avoidable causes' and 'road traffic injuries'. This trend can be observed during the four periods of 5 years. A slowing down of the decrease of the YPLL rates for road traffic injuries, both for men and women is observed: 11.7% between periods 2 and 1, and only 3% between periods 4 and 3 for men (16.2 and 7.5% for women). The geographical analysis shows marked differences between districts. Even though a favourable trend is observed for the traffic injuries deaths in Belgium it is important to highlight the important slowing down of this trend during the most recent years. It is also necessary to underline the importance of geographical disparities in the distribution of YPLL rates within the entire population. PMID- 12081104 TI - Balkan endemic nephropathy in Vratza, Bulgaria, 1964-1987: an epidemiologic analysis of population-based disease registers. AB - Balkan Endemic Nephropathy (BEN), first described in 1956 in Vratza region, Bulgaria, may result from prolonged, chronic exposure to environmental toxicants, but the underlying etiologic factors remain elusive. There has been no recent systematic characterization of the epidemiology of this disease. Recently, it has been suggested that the incidence of the disease is decreasing. We therefore abstracted data from registers of patients in 21 affected villages and the town of Vratza, Bulgaria maintained from 1964 through 1987. In 1964, the prevalence of BEN was 6.0 per 1000 inhabitants; among residents of the affected villages, the prevalence was 12.3 per 1000. From 1965 to 1975 the incidence rate was 0.7 per 1,000 person-years, and from 1976 to 1987 the incidence rate was 0.3 per 1,000 person-years (rate ratio 0.43; p < 0.001). Incidence was much lower in Vratza town; among residents of affected villages, the period-specific rates were 1.7 and 0.8 per 1,000 per year, respectively (rate ratio 0.47; p < 0.01). These trends were consistent across all villages for which registers were maintained. Median survival following registration increased from 2.0 to 5.0 years over the same period (p < 0.001). BEN appears to be decreasing in incidence in this region. PMID- 12081105 TI - Influence of hypertension and smoking as the single vascular risk factors on the intima-media thickness. AB - In order to compare the extent of early carotid atherosclerosis between patients with single vascular risk factors (smoking = SMO or hypertension = HYP) and risk free normal subjects (= HCS), we used intimal-medial wall thickness (IMT) measurements of the common carotid artery. The age- and sex-adjusted mean IMTs were significantly higher in the HYP and SMO groups than in the HCS control group (p < 0.005 and p < 0.001, respectively). The estimated yearly progression rate of IMT (average difference in IMT/year age difference) was significantly higher in the HYP and SMO groups than in the HCS group (p < 0.05). In the total group, the age, hypertension, smoking and sex accounted for 52% of the variability of IMT. Our results show that the estimated yearly progression rate of IMT is slower in healthy and risk-free volunteers than previously detected and that the smoking and hypertension have a similar effect to the progression of early atherosclerosis. This also emphasises the influence of smoking in initial vascular remodelling process. PMID- 12081106 TI - Physical activity and cause-specific mortality in men: further evidence from the Whitehall study. AB - Reports of studies relating physical activity to stroke and cancer sub-types indicate inconsistent findings. Some are hampered by low statistical power, owing to a low number of events, and a failure to adjust for potential confounding variables. The purpose of this study was to relate physical activity to 12 mortality endpoints in a prospective cohort study of 11,663 men aged 40-64 years who responded to an enquiry about travel activity during a baseline medical examination conducted between 1967 and 1969. During 25 years of follow-up there were 4672 deaths. Travel activity was inversely related to mortality attributable to all-causes, coronary heart disease, respiratory disease and lung cancer, whereas the association with stroke was positive. There was evidence for attenuation of some of these associations on adjustment for potentially confounding variables. Our simplistic measure of physical activity may, in part, explain the weak associations seen. PMID- 12081108 TI - Carriers of Neisseria meningitidis in household contacts of meningococcal disease cases in Catalonia (Spain). AB - A population-based study was carried out in Catalonia (Spain) from May 1998 to April 1999 to determine the prevalence of Neisseria meningitidis strains in meningococcal disease (MD) cases and their contacts, as well as the prevalence of meningococci in close contacts of patients with MD, and risk factors for its carriage. A total of 364 close contacts of 87 patients with MD were studied. Throat samples were collected by hospital staff before rifampicin chemoprophylaxis was begun. For each contact, a questionnaire was completed for sociodemographic and epidemiological data. A total of 61 contacts (an overall prevalence of 16.8%) were carriers of meningococcal strains (40 B, 10 C, 1 Z and 10 non-groupable isolates). This prevalence is two to three times higher than in the general population (5-10%). In 33/61 microbiologically confirmed cases (54%) and in 9/26 probable cases (35%), contacts carrying N. meningitidis were found. In 22/33 confirmed cases with carrier contacts, it was possible to study the phenotype of the carrier and patient strains (sero-group, serotype and serosubtype). In 14 cases (64%), both strains were identical, in four cases, only a minor change was observed, in three cases, some strain (from the case or from his contact) was non-serotypable and non-serosubtypable, and in one case, both isolates were completely different. Bivariate analysis identified five statistically significant risk factors for meningococcal carriage: age (5-9 years old), meningococcal A+C vaccination, severe household overcrowding, social class and heavy active smoking (>20 cigarettes a day). Multivariate analysis revealed that of these five variables, only heavy active smoking remained statistically significant when the other factors were controlled. PMID- 12081107 TI - A seroprevalence study of human herpesvirus type 8 (HHV8) in eastern and Central Africa and in the Mediterranean area. AB - Human herpes virus type 8 (HHV8) is the major determinant of Kaposi's sarcoma (KS), a neoplasm with wide geographic variations in incidence rates. To assess the prevalence of HHV8 infection among populations with differing rates of KS, we used sera from 1,402 persons (Central Africa: Cameroon, n = 293, age range: 5-40; eastern Africa: Uganda, n = 315, age range: 1-64: Mediterranean area: Egypt, n = 236, age range: 13-19: Italy, blood donors n = 134, age range: 20-67: Italy. HIV seroconverters n = 424, age range: 16-65). Serum samples were tested for antibodies to lytic and latent antigens of HHV8 using two immunofluorescence assays. HHV8 prevalence was evaluated according to geographic area, gender and age groups. Overall, the highest prevalence of HHV8 lytic antigens (47.5%) was recorded among children and adults in Africa. Approximately 40% of children and adolescents from Egypt and of Italian HIV-positive persons (39.9%) were HHV8 seropositive. In eastern and Central Africa and in Egypt, no differences emerged between males and females for both types of HHV8 antibodies. Conversely, Italian females were at lower HHV8 risk than their male counterparts. Moreover the prevalence of HHV8 infection tended to increase with age. This investigation partially confirms that HHV8 infection mirrors incidence rates of KS. The high prevalence of HHV8 infection in newborns, children and adolescents in Egypt, in eastern and in Central Africa strongly suggests the existence of transmission modes other than sexual. PMID- 12081109 TI - Risk factors of herpes simplex type 1 (HSV-1) infection and lifestyle factors associated with HSV-1 manifestations. AB - This study investigated risk factors for herpes simplex virus type 1 (HSV-1) infection in a population of university students in Germany and Spain. In addition, factors associated with the occurrence of oral lesions were studied. Serum samples were collected from 596 Spanish students from the Navarra Public University in Pamplona and 174 German students from the University of Bielefeld aged 17-41 years and tested by a HSV-1 type-specific immunoassay using monoclonal antibody-selected gG1 as antigen. Information on clinical manifestations and risk factors were obtained by a standardized questionnaire. The rate of HSV-1 infection was 55.3 and 27.4% of these infected students reported having had oral lesions within the last 12 months. Prevalence of HSV-1 infection did not differ between study sites, and did not vary according to gender or age. Students with coitus experience were more likely to be infected with HSV-1 (Odds ratio (OR), 1.88; 95%CI: 1.31-2.69), while other lifestyle factors were not associated with HSV-1 infection. Risk factors for the occurrence of oral lesions included HSV-1 seropositivity (OR: 6.90; 95%CI: 3.84-12.37) and a higher level of perceived stress, as measured by the Cohen scale of perceived stress (OR: 1.79; 95%CI: 1.15 2.78). Drinking alcohol was found to be a protective factor (OR: 0.59; 95%CI: 0.37-0.93). There was no difference in the clinical pattern and course of infection between the young adults in Germany and Spain. We conclude, that lifestyle factors appear to play an important role in the epidemiology and clinical manifestations of HSV-1. PMID- 12081111 TI - Clinical outcome of native valve infective endocarditis in Khon Kaen: 1990-1999. AB - OBJECTIVE: To compare the survival of infective endocarditis (IE) patients following different treatment strategies and to determine the predictors of patient survival. BACKGROUND: IE is a life-threatening infectious disease that is often difficult to manage. Studies on long-term outcome are limited. METHOD: Data on 152 patients with IE from 1990 to 1999 were collected from two hospitals. The main outcome is death after definite diagnosis of native valve IE. RESULTS: The overall case fatality rate was 38 per 100 patient-years. Survival curves showed better survival for patients treated with surgery compared with patients treated medically (p <0.0001). Survival rate at year 1 was 72 per cent for surgically treated patients and 33 per cent for medically treated patients. Five-year survival rates were 66 per cent and 27 per cent in the two groups, respectively. Based on Cox proportional hazards regression analysis, surgery to be an independent predictor of survival (relative risk [RR] = 0.23; 95% confidence interval [CI] 0.14 to 0.39, p < 0.0001), while the presence of congestive heart failure (RR = 2.55; 95% CI 1.61 to 4.02, p < 0.0001), and being male (RR = 1.76; 95% CI 1.04 to 2.82, p <0.05) were independent predictors of mortality. CONCLUSION: Patients with native valve endocarditis have a high long-term mortality rate. The most common types of cardiac death are post-operative and sudden death. Surgical treatment was the preventive factor of mortality. PMID- 12081110 TI - Seroprevalence of Borrelia burgdorferi in Romania. AB - In order to assess the seroprevalence of Borrelia burgdorferi in Romania and to define associated risk factors, a cross-sectional, observational study was performed in 13 districts during 1999. Sera from healthy blood donors (1,598) and from forestry workers (1,048) were tested. A two-step testing strategy was used in which sera were tested for anti-B. burgdorferi antibodies by a commercially available passive hemagglutination assays (PHA). All PHA positive sera were then evaluated by Western blot IgG. Demographic data regarding age, sex, profession, work place/residence, duration of employment (forestry workers), animals in the environment, and tick bites history were collected using a questionnaire. Data obtained from serological study were matched with that obtained from the questionnaire. The seroprevalence of B. burgdorferi in blood donors was of 4.3% (range 1.4-8.7%) and 9.3% (range 2.8-31.7%) in forestry workers. Seroprevalence was higher in forestry workers with a tick bite history (10.7 vs. 4.3%, p < 0.05). The highest seroprevalence in blood donors (8.7%) was noted in Maramures, a northern district of the country, whereas in forestry workers the highest seroprevalence (31.7%) was observed in a western district (Arad), where a previous study in entomology has demonstrated the highest density of Ixodes ricinus ticks in Romania. PMID- 12081112 TI - Evaluation of screening tests and prevalence of glaucoma: integrated health research program for the Thai elderly. AB - The purposes of this study were to evaluate the accuracy of glaucoma screening tests and to assess the prevalence of glaucoma in an elderly Thai population over 60 years of age. This was a cross-sectional study. 3706 subjects were recruited from 33 communities in the vicinity of Siriraj Hospital. All received an eye examination from ophthalmologists and general practitioners including visual acuity, Schiotz tonometry and optic disc evaluation in their residential communities. Subjects who were suspected of having glaucoma were re-examined at the Glaucoma Clinic, Siriraj Hospital. They were then classified as having definite glaucoma of a specific type or as not having glaucoma. The overall prevalence of glaucoma was 6.1 per cent (128/2092). The percentages of primary open angle glaucoma, primary angle closure glaucoma, normotension glaucoma and secondary glaucoma were 47.7 per cent, 41.4 per cent, 9.4 per cent, and 1.6 per cent respectively. The sensitivity and specificity of the screening tests performed by ophthalmologists were 100 per cent and 97.2 per cent and those performed by general practitioners were 21.7 per cent and 96.5 per cent. PMID- 12081113 TI - The epidemiology of osteoarthritis of the knee in elderly patients living an urban area of Bangkok. AB - An epidemiological study of 392 elderly Thai patients with osteoarthritis (OA) of the knee, 86 males and 306 females, with a mean age of 67.8 years, was performed. The prevalence of knee OA was 34.5-45.6 per cent. Most of the patients had completed primary education and had sufficient income. More than half of the patients had pain in both knees, but many of the elderly people used neither knee supports nor walking aids (82.7 and 90.6%). Sixty-four per cent had no previous knowledge concerning the value of exercise. Sixty-five per cent of the patients had never used any pain medication. Three-fourths of the subjects evaluated themselves to have fair to good health. Sixty per cent presented with knee pain when climbing the stairs, and 64.8 per cent had a fair quality of life. Almost all the subjects had mild to moderately severe grade of OA and their X-ray findings were level II-III. The average quadriceps strength and six-minute walking distance were 12.95 +/- 5.51 kilograms and 363.69 +/- 99.34 meters respectively. The five factors that significantly correlated with walking ability were age, sex, the functional incapacity score, using walking aids and level of severity found on X-ray. Previous knowledge about knee exercise did not affect walking ability, but may have been due to the irregularity of the exercise schedule they performed. The elderly should be encouraged to exercise regularly in addition to receive knowledge about proper exercise. PMID- 12081114 TI - Relationship between corneal thickness and level of myopia. AB - A retrospective study of 533 eyes, which underwent complete pre-operative evaluation for refractive surgery, was done. Regression Analysis was performed to find the correlation between corneal thickness and level of myopia and between corneal thickness and corneal curvature. There was statistically significant correlation between corneal thickness and level of myopia (p = 0.039) and also in corneal thickness and corneal curvature (p = 0.04). No clinical correlation was demonstrated (R2 = 0.014 and R2 = 0.0153, respectively). There was no clinical correlation between corneal thickness and level of myopia and also between corneal thickness and corneal curvature. PMID- 12081115 TI - Knowledge-attitude-practice of sexual intercourse of post-menopausal women using hormone replacement therapy. AB - OBJECTIVE: To study knowledge, attitude, practice and concern related to sexual intercourse of post-menopausal women using hormone replacement therapy. MATERIAL AND METHOD: One hundred and thirty subjects were included in the study from the 1st of May 2000 to the 31st of August 2000 at the Menopause Clinic, Srinagarind Hospital, Khon Kaen. All subjects were interviewed by a researcher with structured questionnaires. RESULTS: Sixty seven per cent of the subjects had an acceptable level of knowledge, whereas, 87.7 per cent thought that they could have sexual intercourse similar to what they had in their pre-menopausal period. It was found that 85.4 per cent and 88.5 per cent of the subjects had either less or absent sexual desire and less sexual activity, respectively. In terms of their concern, 73.9 per cent of the subjects were not worried about sexual intercourse. CONCLUSION: The majority of the post-menopausal women using hormone replacement therapy had a good level of knowledge and attitude concerning sexual intercourse. Although most of them had decreased sexual desire and activity, this sexual activity did not cause them concern. PMID- 12081116 TI - Laparoscopic cholecystectomy versus open cholecystectomy in children. AB - INTRODUCTION: Laparoscopic cholecystectomy (LC) is well accepted as the standard cholecystectomy only in adult patients. However, the advantages of LC over open cholecystectomy have never been proved in pediatric patients because the number of pediatric cholecystectomies is limited as well as the faster ability of pediatric patients to resume their normal activity. MATERIAL AND METHOD: Retrospective study of 42 pediatric cholecystectomies (laparoscopic cholecystectomy (n = 8) (LCs), open cholecystectomy alone (n = 8) (OCs) and open cholecystectomy concomitant with splenectomy (n = 26)(OCs + S)) done in Siriraj University Hospital, Bangkok, Thailand between 1992 and 2000 was conducted. RESULTS: Statistical comparison revealed that LC was superior to OC in regard to diet resumption. LCs resumed soft diet on 1.38 days, whereas OCs and OCs + S could resume soft diet on 3.38 and 3.35 days respectively. The average length of hospitalization following LCs was significantly shorter than OCs' and OCs + S' ones (3.00 vs 8.38 and 4.85 days respectively). There was no morbidity and mortality in LCs, whereas two OCs and three OCs + S had complications. CONCLUSION: In this preliminary study, laparoscopic cholecystectomy is a preferred method of cholecystectomy in children because it has a shorter post operative interval of diet resumption and shortens hospitalization with minimal morbidity. However, this study has a limited number of patients and further study is still required to conclude the benefits of LC. PMID- 12081117 TI - Penile fracture: long-term outcome of treatment. AB - OBJECTIVE: Penile fracture is a rare injury, usually resulting from direct trauma to the erect penis during sexual intercourse. Immediate surgical treatment is the basis for the treatment of this injury due to the high rate of complications associated with delayed management. The aim of this study was to evaluate the clinical presentations, diagnostic methods, and outcomes of the treatment. MATERIAL AND METHOD: We retrospectively studied patients with penile fracture treated at Ramathibodi Hospital from 1975 to 2000. Clinical presentation, diagnostic methods, technique of treatment and outcomes of treatment were noted. RESULTS: Twelve patients were found in this study. The mean patient age was 32 years old (range 19-42). The interval from time of injury to presentation was 3 48 hours. Of these patients 10 had been injured during sexual intercourse (83%) while 2 had been injured during masturbation. All patients presented with a very suggestive clinical picture (pain, detumescence and hematoma). No further investigation was needed for confirming the diagnosis. One case had urethral bleeding, therefore, retrograde urethrogahpy was performed but no extravasation of contrast media was noted. All patients were treated by immediate surgery, through a circular subcoronal incision and degloving of the penis to allow a thorough exploration. All of the patients had a tunica albuginea tear that was promptly repaired. No associated urethral larceration was noted. All of the patients did very well after surgery and two had mild curvature, which had not hindered intercourse at follow-up (mean time of 24 months). CONCLUSIONS: Penile fracture has very typical clinical signs and no further investigation is usually needed. Early surgical treatment is associated with a low incidence of late complications. PMID- 12081118 TI - Epidemiological correlation between chromium content in gallstones and cholesterol in blood. AB - The authors measured the chromium in gallstones and bile from patients in three areas (Kawasaki (a city adjacent to Tokyo) in Japan, Chiang Mai and Bangkok in Thailand) by means of neutron activation analysis. The chromium in three types of gallstones (cholesterol, pigment, and rare stones) and bile from patients living in Bangkok were evidently larger than those from patients living in Kawasaki and Chiang Mai. The high chromium intake by Bangkok patients continued from the start of gallstone formation until the time the stones were removed. The total cholesterol, triglyceride, and hemoglobin A(1C) levels in the blood from Bangkok residents with high chromium intake over a long period were clearly lower than those of Japanese and Chiang Mai residents. The authors showed that the high dietary intake of chromium over a long period may play a role in the lowering of total-cholesterol, triglyceride, and hemoglobin A(1C) in blood. PMID- 12081119 TI - Efficacy of the ramathibodi nasal filter in a simulated human airway evaluated by a laser diode portable dust monitor under low laser smoke particle concentration. AB - The Ramathibodi nasal filter attached to a simulated human airway was proposed to filter laser smoke particles. The simulated human airway composed of nasal and pharyngeal model, airway passage and lung model machine which mimicked the human respiratory system. The laser smoke particles represented a suspended particulate matter in a highly air-pollutted area such as at a main roadside in Bangkok. The experiment was done in the Department of Otolaryngology, Ramathibodi Hospital, from January to March 2000. The simulated human airway got an equal amount of laser smoke particles in a sealed plastic box for 1 min. The residual amount of laser smoke particles in a closed system of the simulated human airway was measured by a laser diode portable dust monitor for 1 min in each cycle and calculated as a mean and standard deviation. Without the Ramathibodi nasal filter of 39 sample pairs, the amounts of PM15, PM10 and PM2.5 were 52.3 +/- 6.8, 43.0 +/- 4.9 and 37.0 +/- 3.5 mcg/m3 respectively. With the Ramathibodi nasal filter of 39 pairs sample, the amounts of PM15, PM10 and PM2.5 were 48.1 +/- 9.9, 39.1 +/- 9.1 and 33.2 +/- 7.2 mcg/m3 respectively. Ramathibodi nasal filter efficacy for all laser smoke particle sizes evaluated statistically using t-test showed significant differences from those without the filter. Filtration efficacy should be tested further in higher concentrations of laser smoke particles and applied in human nasal vestibules under a critical air-polluted condition. PMID- 12081120 TI - Pelvic fractures: experience in management of 170 cases at a university hospital in Thailand. AB - BACKGROUND, OBJECTIVE AND METHOD: Management of patients with pelvic fractures requires a multidisciplinary team approach. Currently, survival has been dramatically improved but some controversies still remain. The purpose of this study was to examine management and results of treatment of patients with pelvic fractures who were admitted to the Trauma Unit, King Chulalongkorn Memorial Hospital, Bangkok, Thailand from January 1991 to December 2000. RESULTS: There were 170 patients in the study. The age ranged from 15 to 91 years (mean 33.89 +/ 16.14). The most common cause of injuries was motorcycle accidents (50.0%). There were 27 (15.9%), 47 (27.6%), 80 (47.1%) and 16 (9.4%) patients with Type I, II, III and IV pelvic fractures, respectively. Forty per cent of patients were in shock when they first arrived at the emergency room. Seventy two patients (42.4%) had 274 associated injuries. Sixteen patients (9.4%) had open pelvic fractures. The Injury Severity Score (ISS) ranged from 4 to 75 (mean 17.55 +/- 12.86). Eighty two patients (48.2%) received blood transfusion from 1 to 40 units (mean 10.04 +/- 8.47). Sixteen patients (9.4%) underwent pelvic angiography, 10 bleeding points were demonstrated and successfully treated by transcatheter embolization. One hundred and thirty two patients (77.6%) received no specific treatment for the pelvic fractures. The remainder (22.4%) were treated with pelvic sling in 6 patients (3.5%), skeletal traction in 21 patients (12.4%), external fixation in 6 patients (3.5%), internal fixation in 4 patients (2.4%), and right hemipelvectomy in 1 patient (0.6%). Fifteen patients (8.8%) died. Causes of death were exsanguination in 6 patients (40% of death), severe head injuries in 6 patients (40% of death) and sepsis with multisystem organ failure in 3 patients (20% of death). Nonsurvivors had a significantly higher ISS and units of blood transfusion than survivors (P < 0.001). The hospital stay ranged from 1 to 300 days (mean 24.7 +/- 34.19). CONCLUSION: Approximately 75 per cent of patients in our study had major pelvic fractures (Type II and Type III pelvic fractures). The majority of bleeding from pelvic fractures could be treated conservatively. Angiography with transcatheter embolization was extremely helpful when conservative treatment failed to stop pelvic bleeding. External fixation for early control of bleeding pelvic fractures was infrequently employed. PMID- 12081121 TI - Prevention of phantom sensations after spinal anesthesia. AB - BACKGROUND AND OBJECTIVES: Phantom limb sensation (PLS) after the spinal anesthesia is an unpleasant experience. The occurrence rate was 80 to 83 per cent from previous studies. The purpose of this study is to evaluate the incidence of PLS, comparing between placing the lower limbs in the neutral position when analgesic level reach L1 dermatome (L1 group) and T6 dermatome (T6 group). METHODS: Ninety ASA physical status I-II patients, 19-50 year of age, scheduled for lower abdominal surgery under spinal anesthesia were enrolled. Immediately after the spinal injection, the patients were placed supine with one of lower limbs flexed (both hip and knee) and the other kept straight in the neutral position. The analgesic levels were checked every minute; the flexed limbs were turned to the neutral position when the analgesic level reached L1 and T6, the L1 group and the T6 group respectively. The images of the lower limbs and patients' satisfactions were evaluated 15 minutes after the blockage. RESULTS: Twenty per cent of the patients in the L1 group experienced PLS of the flexed limbs while 82.2 per cent of the patients in the T6 group did (P<0.05). None of the patients in the L1 group felt unsatisfied with PLS, on the other hand, 11.1 per cent of the patients in the T6 group expressed their dissatisfaction and would like to have their limbs extended. CONCLUSIONS: This study revealed that by placing the lower limbs in the neutral position immediately after the spinal injection, before the unwanted positions are memorized, could effectively minimize phantom limb sensation after spinal anesthesia. PMID- 12081122 TI - Strategy to prevent falls in the Thai elderly: a controlled study integrated health research program for the Thai elderly. AB - Instability or falls are one of the important warning symptoms of underlying serious illness among the elderly, so many studies have concentrated on the risk factors for falls. However, a study involving a strategic method to reduce the incidence of falls is the next step and is the main objective of this study. 1,043 elderly subjects living in the urban area around Siriraj Hospital Medical School, Bangkok, were recruited, 585 of them were allocated to the study group and 458 subjects to the control group. A leaflet containing information on important risk factors of falls within their community was enclosed with a follow up postcard in the study group only. In addition, this particular group was allowed free access to the geriatric clinic at Siriraj Hospital if there was any health problem. All of them received a postcard asking about any falls which had occurred over the previous 2 months on 6 occasions and a telephone call if the postcards were not returned to the team. The percentage of elderly who kept in contact was 92.5 per cent, 90.6 per cent, 89.3 per cent, 89.2 per cent, 86.2 per cent and 85.45 per cent for the first to final follow-up respectively. After one year of longitudinal study, the overall incidence of falls was 6.6 per cent in the study group and 10.1 per cent in the control group. The incidence of falls began to show a statistically significant difference between the two groups at the fourth and sixth episodes of follow-up (P = 0.002 and 0.004). The Kaplan Meier survival analysis also revealed a statistically significant difference in the incidence of falls between the two groups (P =0.01). In conclusion, the incidence of falls was significantly reduced in the study group and a repeated campaign to alert the elderly to the risk of falling is a cost-effective way of fall prevention among the healthy elderly in the community. PMID- 12081123 TI - Evaluation of transurethal alprostadil for safety and efficacy in men with erectile dysfunction. AB - A multicenter study conducted in 5 trial centers, for the safety and efficacy of transurethral alprostadil (Pellet) in 90 subjects. The end results show quite good and the product was satisfactory and safe for all ages. Although there are multiple side effects that seemed to discourage the participants and caused them to dropout, MUSE does work but needs good understanding of the use of the device. MUSE is another alternatives for those who unable to use oral medication or those who require immediate action but need some dexterity. PMID- 12081124 TI - Antiseptics for preventing omphalitis. AB - BACKGROUND: Omphalitis may cause serious complications and contribute to neonatal morbidity and mortality. From January 1997 to August 1998, the incidence of omphalitis in the Department of Obstetrics and Gynecology, Faculty of Medicine Siriraj Hospital had been increased from 0.9 to 17.4 per 1,000 live births. A prospective randomized trial using antiseptic applied directly to the umbilical stump was conducted aiming to reduce an epidemic outbreak of omphalitis in the newborn nursery. OBJECTIVE: To determine which antiseptic is appropriate for preventing omphalitis in the newborn infants. PATIENTS AND METHOD: Newborn infants delivered in the Department of Obstetrics and Gynecology, Faculty of Medicine Siriraj Hospital were randomized into group A (Triple dye) or group B (70% Alcohol). The infant with omphalitis was assessed by a pediatrician or a neonatology fellow. At home, the same antiseptic will be continually applied to the umbilical stump daily until a few days after cord detachment. Relative risk was calculated and statistical significance was tested by Chi-square test. RESULTS: Four hundred and twenty-seven infants were enrolled. Birth weight, gestational age and gender of the infants in both groups were not different. There were no known maternal risk factors for omphalitis. Omphalitis was observed in 9/213 (4.2%) infants in group A and 23/214 (10.7%) infants in group B. The relative incidence rate between each group was statistically significant (p<0.01). Triple dye group was 60 per cent less likely to develop omphalitis compared to 70 per cent Alcohol group (RR 0.39, 95% CI: 0.19-0.83). The mean duration for cord detachment were 13.6 and 11.5 days in group A and group B, respectively. CONCLUSION: During an epidemic outbreak of omphalitis, Triple dye was the most appropriate and effective antiseptic to prevent omphalitis but could delay cord separation. PMID- 12081126 TI - Nuchal translucency in normal fetuses in a Thai population. AB - The purpose was to study nuchal translucency thickness of normal Thai fetuses in the first trimester. From January 1996 to June 1999, there were 1,970 pregnant women, whose gestational ages were between 10 to 13(+6) weeks of gestation, who attended the Down syndrome screening clinic. Nuchal translucency (NT) thickness measurement was used as the screening method for Down syndrome. When the result showed an increased risk for Down syndrome, the patient was counselled and the invasive test for the definite diagnosis was optional. Pregnant women who had had an abortion and/or congenital anomaly babies were excluded. Only pregnant women who had normal newborns were recruited in this study. There were 1,631 pregnant women who had normal newborns in this study. The mean maternal age was 28.70 +/- 5.14 years. There were 222 (13.6%) pregnant women whose ages were more than or equal to 35 years. There was quadratic correlation of an increase in nuchal translucency and the crown-rump length (CRL) in normal fetuses in the first trimester. The quadratic equation was log10 NT = 1,215 x 10(-4) CRL-8.7 x 10(-4) CRL(2)-3.7037. The normal fetuses with nuchal translucency thickness more than or equal to 2.5 mm were found in 1.23 per cent in this study. In conclusion, there was quadratic correlation of nuchal translucency thickness and crown-rump length of first trimester fetuses in a Thai population. PMID- 12081125 TI - Bone mineral density in rural Thai adults living in Khon Kaen province. AB - OBJECTIVES: To define the bone status and pattern of bone loss in a normal adult population living in a rural area of Khon Kaen province. STUDY DESIGN: A descriptive study. SETTINGS: Department of Medicine, Faculty of Medicine, Khon Kaen University, Thailand. Subjects 436 subjects (181 men and 255 women), aged 20 80 years. MAIN OUTCOME MEASURES: Bone parameter including total bone mineral density (BMD), the bone mineral density at different sites such as forearm, lumbar spine and hip, bone mineral content (BMC) were measured by dual energy X ray absorptiometry (DEXA). RESULTS: The result revealed that women had lower bone mass at various bone sites than men. The peak bone mass in women was observed between 30-39 years of age except at the distal radius which occurred between 20 29 years of age while the peak bone mass in men was observed between 20-29 years of age. It was also shown that bone loss occurred in both men and women with advancing age. The rate of decline at all sites in women was greater than men especially when they were over 60 years old. When comparisons were made between pre and post-menopausal women, the mean of bone mineral density in pre-menopausal women was 1.18 +/- 0.08, 0.69 +/- 0.06, 0.69 +/- 0.06, 1.0 +/- 0.13, 1.15 +/- 0.13 g/cm2 at total body, mid-shaft radius, ultra distal radius, femoral neck and lumbar spine, respectively while the mean of bone mineral density in post menopausal women was 1.02 +/- 0.12, 0.54 +/- 0.11, 0.54 +/- 0.11, 0.75 +/- 0.16, 0.88 +/- 0.2 g/cm2 at total body, mid-shaft radius, ultra distal radius, femoral neck and lumbar spine, respectively which were lower than BMD in pre-menopausal women (p<0.05). The mean bone mineral content (BMC) in pre-menopausal women was 2401+318.3 g while in post-menopausal women it was 1915.4+421.7 g (p<0.05). The rate of bone loss correlated with the duration after menopause. In this study, using World Health Organization criteria for diagnosis of osteopenia and osteoporosis and a reference value obtained from Khon Kaen young adults, the prevalence of osteopenia in Khon Kaen women subjects was 37.4 per cent at femoral neck, 30.2 per cent at lumbar spine, 44.5 per cent at ultra distal radius, 31.5 per cent at mid-shaft radius and the prevalence of was osteoporosis 19.3 per cent at femoral neck, 24.7 per cent at lumbar spine, 18.5 per cent at ultra distal radius and 26.4 per cent at mid-shaft radius. CONCLUSION: The result of this study demonstrated the bone parameters in rural Thai adults living in Khon Kaen province, the pattern of bone loss, the difference between men and women and finally the prevalence of osteopenia and osteoporosis. PMID- 12081127 TI - Congenital megacolon: diagnosis using size of sub-mucosal nerve trunk as a criteria. AB - Cholinergic hyperinnervation in submucosa of aganglionic segment of Hirschsprung's disease has been described. However, objective measurement of the nerve fibers was insufficiently reported. OBJECTIVES: To study the amount and size of hypertrophic submucosal nerve trunks in the aganglionic segment of Hirschsprung's disease, semiquantitatively and evaluate the possibility of using the size as objective diagnostic criteria. MATERIAL AND METHOD: Thirteen specimens from aganglionic segments from Hirschsprung's disease and six specimens from the age-matched control subjected without colonic innervation disorders were studied with the S-100 immunohistochemical technic. The submucosal nerve trunks were counted per ten HPF and measured in width, semiquantitatively. Fiber sizes were stratified into small (<20 micrometers), medium (20-40 micrometers), and large (>40 micrometers). Maximum fiber size in each specimen was also recorded. RESULTS: Average density of submucosal nerve trunk in the aganglionic segment and the control were 12.5 and 3.4 fibers per ten HPF, respectively. The large nerve trunks were detected in 12 of 13 aganglionic specimens and none was detected in the controls. The maximum fiber size in aganglionic segments was 78.2 micrometers, compared to 24.1 micrometers in the control group. The sensitivity and specificity to set the presence of hypertrophic nerve trunk, larger than 40 micrometers as a diagnostic criteria for Hirschsprung's disease were 92.3 and 100 per cent, respectively. The data suggest the use of this size as an objective diagnostic criterion, which may be valuable in rectal suction biopsy specimens. PMID- 12081128 TI - Unenhanced ultrafast computerized tomography for the evaluation of patients with acute flank pain. AB - OBJECTIVE: To determine the value of unenhanced ultrafast computerized tomography (CT) in the diagnosis of acute flank pain in 43 patients evaluated for suspected stone disease. MATERIAL AND METHOD: Noncontrasted ultrafast CT was performed in 43 consecutive patients seen in the emergency department to evaluate acute flank pain. All CT studies were reviewed for the presence of ureteral and renal calculi, perinephric and periureteral stranding, presence and degree of pelvicalicectasis or other radiological findings. If necessary, an excretory urogram was performed to confirm the presence or absence of urinary stone. Patients were followed to determine clinical outcome including the need for urological intervention. RESULTS: Of the 28 patients determined to have stones 16 (57.14%) had spontaneous stone passage, 7 (25%) had improved symptoms without documented stone passage and 4 (14.29%) required surgical intervention. In 6 of 14 patients (42.86%) with negative CT readings for stone disease a diagnosis was established by other intra-abdominal findings. In 7 patients (50%) no clinical diagnosis could be established, and 1 scan in a patient with a ureteral calculus was interpreted as falsely negative. These findings yielded a sensitivity of 96.63 per cent, Specificity 92.85 per cent and overall accuracy 95.24 per cent for diagnosing ureteral stones. CONCLUSIONS: Unenhanced ultrafast CT is an accurate, safe and rapid imaging modality for the detection of urinary tract calculi and obstruction. The majority of patients required no further imaging to determine the need for urological intervention. Ultrafast CT could be used as the standard method to evaluate patients with acute flank pain. PMID- 12081129 TI - Drug eruptions: the value of oral rechallenge test and patch test. AB - All in-and out-patients, who came for drug eruption consultation at the Dermatology Clinic, Ramathibodi Hospital from December 1997 to November 1998 were included in this study. Medical histories and physical examinations were performed by one of the authors. In suspected cases, a skin biopsy was performed to confirm the diagnosis. Patch test and oral challenge test were performed in some patients who had maculopapular, fixed drug eruption and acute generalized exanthematous pustulosis, with informed consent. Among 80 patients, the most common cutaneous reaction was maculopapular rash. Antimicrobial, drugs were the most common causative agents. The patch test was positive in only one patient from 12 cases. The oral provocative test was positive in two patients from 4 cases. It is concluded that oral provocative test is still necessary to get a definite diagnosis of causative agent. The value of patch test needs further study. PMID- 12081130 TI - Carcinoma of the breast with unusual clinical presentation: a case report. AB - A common presentation of breast cancer is a palpable or non-palpable breast mass that might or might not demonstrate clinical signs of skin and/or nipple involvement. These malignant lesions frequently receive correct diagnosis only by physical examination under modestly experienced observers. We, hereby, reported the case of a 66-year-old woman who presented with a rapidly growing skin lesion at the left breast for three months. A 3.6x3x2.5 cm reddish, shiny, rubbery, sessile mass was seen on the upper outer quadrant of her left breast. The cytologic and histologic features of the tumor were typical for infiltrating ductal carcinoma. This case demonstrated a very unusual clinical presentation of breast cancer. No similar clinical feature of such cancer was mentioned in the reviewed literature. PMID- 12081131 TI - Medical ethics and infertility. PMID- 12081132 TI - Practical aspects of the ligand-binding and enzymatic properties of human serum albumin. AB - Recent work with approaches like recombinant mutants and X-ray crystallography has given much new information about the ligand-binding properties of human serum albumin (HSA). The information increases the understanding of this unique transport and depot protein and could give a structural basis for the possible construction of therapeutic agents with altered HSA-binding properties. A tabulation of high-affinity binding sites for both endogenous and exogenous compounds has been made; it could be useful for the above-mentioned purpose, but it could also be of value when trying to predict potential drug interactions at the protein-binding level. Drug displacement is not always a complication to therapy; it can be used to increase the biological effect of a drug. However, due to rebinding at other sites, the increase in the free concentration of a displaced ligand can be less than expected. Drugs and drug metabolites can also interact covalently with HSA; thiol-containing drugs often bind to the single free cysteine residue of HSA, and glucuronidated drugs react irreversibly with other residues of the protein. Reversible binding of ligands is often stereospecific, and therefore immobilized HSA can be used to separate drug isomers. Albumin-containing dialysates are useful for extracorporeal removal of endogenous toxins and in the treatment of drug overdoses. HSA has different types of hydrolytic activities, which also can be stereospecific. The esterase-like property seems especially useful in converting prodrugs to active drugs in plasma. PMID- 12081133 TI - cDNA cloning of the Sry-related gene Sox6 from rat with tissue-specific expression. AB - A cDNA encoding rat homologue of the previously characterized mouse Sox6 was isolated by a polymerase chain reaction (PCR) cloning strategy. Comparison of this eDNA with homologous mouse, human and rainbow trout cDNA exhibited an overall amino acid sequence identity of 99.6, 89.3 and 76.3% respectively. The leucine-zipper and HMG-box motif were almost completely conserved between these homologues. The expression of Sox6 was determined in rat by Northern hybridization and Real-time quantitative reverse transcription (RT)-PCR. rSox6 (rat Sox6) was specifically expressed in the neonatal brain and adult testis with Northern blotting. Real-time quantitative RT-PCR for the determination of Sox6 mRNA was examined. The rSox6 was expressed in the neonatal brain and adult testis as well as by Northern blotting and also expressed in the adult eyeball and slightly in the ovary. PMID- 12081134 TI - Participation of the arachidonic acid cascade pathway in macrophage binding/uptake of oxidized low density lipoprotein. AB - Arachidonic acid cascade inhibitors, including phospholipase A2 inhibitors, dexamethasone and quinacrine (mepacrine), cyclooxygenase inhibitors, indomethacin and aspirin, and lipoxygenase inhibitor AA861, prevented foam cell formation and cholesterol accumulation in the incubation of thioglycollate-induced mouse peritoneal macrophages with oxidized low density lipoprotein (LDL) at 37 degrees C for 24 h. These inhibitors similarly prevented foam cell formation of fibronectin- and Ca ionophore A23187-stimulated macrophages. Binding and/or uptake of Dil (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine)-acetyl LDL by macrophages at 37 degrees C for 3h and arachidonic acid release from macrophages at 37 degrees C for 4h were inhibited by dexamethasone. Exogenously added phospholipase A2 of bee venom and Crotalus adamanteous venom increased arachidonic acid release during incubation for 2 h, and increased macrophage binding and/or uptake of Dil-acetyl LDL at 37 degrees C for 3 h, and foam cell formation at 37 degrees C for 24 h. Protein kinase inhibitors, ML-9 and staurosporine, that inhibited macrophage binding and/or uptake of Dil-acetyl LDL did not inhibit arachidonic acid release, indicating that protein phosphorylation was not involved in the arachidonic acid pathway in the macrophage scavenger receptor activation. Nordihydroguaiaretic acid that inhibited arachidonic acid release prevented binding and/or uptake of Dil-acetyl LDL. The release of arachidonic acid was not enhanced by fibronectin-stimulation, indicating that Ca influx-dependent stimulation of macrophage activity was not through the activation of phospholipase A2. These results indicate that, as well as the fibronectin-stimulated Ca influx pathway and protein phosphorylation pathway, the arachidonic acid pathway participated in the activation of macrophages to bind and take up oxidized LDL. PMID- 12081135 TI - Evidence of autophosphorylation in Txk: Y91 is an autophosphorylation site. AB - We have previously shown that Txk, a member of Tec family tyrosine kinase, is expressed in Th1 and ThO cells and directly contributes to gene transcription of Th1-related proteins, including interferon (IFN)-gamma, through nuclear translocation in response to mitogenic stimuli. Btk, another member of Tec family tyrosine kinase, has been shown to have a Src family tyrosine kinase-dependent transphosphorylation site and an autophosphorylation site. However, little is known about the phosphorylation mechanism of Txk, except that 420 tyrosine residue was identified as the transphosphorylation site. In this study, we found that Txk autophosphorylated itself by using an in vitro kinase assay. To elucidate the role of phosphorylation in Txk function, we studied IFN-gamma secretion by Jurkat T cells expressing mutant Txk proteins. While transfection with the wild-type Txk resulted in increased IFN-gamma production, the function was abrogated by disruption of the ATP biding site, which is presumably involved in the autophosphorylation mechanism. The results suggest that phosphorylated Txk is an active form to promote IFN-gamma synthesis. The 91 tyrosine residue of Txk is deduced to be an autophosphorylation site by comparing its structure with Btk. In Jurkat cells transfected with Txk Y91A, IFN-gamma production was decreased in comparison with the wild-type Txk transfected Jurkat cells. These data suggest that phosphorylation of the 91 tyrosine residue in Txk plays a positive regulatory role in Txk function. PMID- 12081136 TI - Effect of replacing the aspartic acid/glutamic acid residues of bullfrog sialic acid binding lectin with asparagine/glutamine and arginine on the inhibition of cell proliferation in murine leukemia P388 cells. AB - The sialic acid binding lectin from bullfrog oocytes (cSBL) is known to have anti tumor activity. In a previous report, to elucidate the relationship between the net charge and anti-tumor activity of cSBL, we examined the effect of chemical modifications of cSBL with a water-soluble carbodiimide in the presence of various nucleophiles. The results suggested that the anti-tumor activity and internalization into tumor cells increased with an increase in the net charge of cSBL. However, in the chemically modified cSBL, a modification site was observed on average in two of the carboxyl groups of cSBL. To confirm these previous results and to determine which modified carboxyl group contributes to the increase in anti-tumor activity, we prepared mutants with substitutions of Asn/Gln and Arg at three acidic amino acid residues of cSBL and studied their anti-tumor activity and internalization efficiency. The results showed the enhancing effect of charge on anti-tumor activity and internalization, and suggested that the replacement of D24 and E88 of cSBL with arginine is more effective than that of E97. The double mutant D24RE88R showed comparable anti tumor activity to the ethylenediamine-modified cSBL reported previously. The mutant was well-characterized as a pure cSBL derivative suitable for studying the mechanism of the anti-tumor action of cSBL. PMID- 12081137 TI - G1-dependent prion protein expression in human glioblastoma cell line T98G. AB - Human glioblastoma cell line T98G produced a cellular form of prion protein (PrP(C)), and we confirmed expression of PrP mRNA by RT-PCR. Immunoblot analysis of whole cell lysate revealed one major (35 kDa) and two faint bands (31, 25 kDa) that reacted with monoclonal anti-human PrP antibody 3F4. Cells treated with tunicamycin produced only a 25 kDa band, representing a deglycosylated form of PrP. Similarly, peptide: N-glycosidase F treatment of whole cell lysate altered the Asn-linked form to the deglycosylated form. When T98G cells were cultured for a longer period, the amount of PrP(C) per cell increased on Day 4 to 16 in a time dependent manner. When the cells were cultured at high cell-density, the cells on Day 4 produced the same amount of PrP(C) as those on Day 16 of the usual culture. Moreover, in a serum-free medium, cells cultured at a low cell-density produced the same amount of PrP(C) as those cultured at the high cell-density. These results demonstrate that PrP(C) production in T98G cells was dependent on the phase of the cell cycle, probably the G1 phase. PMID- 12081138 TI - Comparison of subcellular distribution of mevalonate pyrophosphate decarboxylase between stroke-prone spontaneously hypertensive rat and Wistar Kyoto rat. AB - We previously reported that the lower activity of mevalonate pyrophosphate decarboxylase (MPD) was caused by the reduced amount of this enzyme in stroke prone spontaneously hypertensive rat (SHRSP) by immunoblot analysis using 20,000 x g supernatant containing cytosol and microsomes. A recent study showed that at least three different subcellular compartments, including peroxisomes, are involved in cholesterol synthesis. In this study, we examined the subcellular distribution of 45- and 37-kDa MPD in the liver of SHRSP and compared normotensive Wistar Kyoto rat (WKY) and SHRSP. 45-kDa MPD was detected in the cytosol and peroxisomes of SHRSP, while 37-kDa MPD was detected in the cytosol of SHRSP, but not in the peroxisomes. The relative enrichment of 45-kDa MPD in peroxisomes was lower than that of LDH, suggesting the possibility that 45-kDa MPD of SHRSP did not exist in the peroxisomes. Also, 45-kDa MPD was decreased in the crude extract containing 1% Triton X-100, cytosol and peroxisomes of SHRSP, and 37-kDa MPD was decreased in the crude extract containing 1% Triton X-100 and cytosol of SHRSP, as compared with WKY. These data indicate that the cholesterol synthesis in the liver of SHRSP by the reduced amount of MPD is significantly reduced. PMID- 12081139 TI - Effects of osthole on postmenopausal osteoporosis using ovariectomized rats; comparison to the effects of estradiol. AB - The purpose of this study was to examine effects of osthole on postmenopausal osteoporosis using ovariectomized (OVX) rats. All of the rats were divided into sham and OVX groups. At 2 weeks post-operation, the sham-operated rats received solvent vehicle (97% corn oil and 3% ethanol, 1.0 ml/kg, subcutaneously); the OVX rats were divided into three groups which were treated with solvent vehicle (same the sham rats, 1.0 ml/kg, subcutaneously), 17beta-estradiol (30 microg/kg, subcutaneously) or osthole (9.0 mg/kg, orally) 5 d/week for 4 weeks, respectively. In OVX rats, the increases of body weight, spleen and thymus weight were significantly decreased and the atrophy of uterus was preserved by 17beta estradiol treatment, but not by osthole. Treatment with either 17beta-estradiol or osthole significantly protected cancellous bone loss owing to estrogen deficiency and significantly increased the maximal load in the femoral neck of OVX rats. In addition, the increases of serum osteocalcin (OC) and urinary deoxypyridinoline (DPD) levels caused by ovariectomy were all significantly suppressed by 17beta-estradiol. However, only urinary DPD was significantly reduced by osthole and no change was found in serum OC. Our results demonstrate that osthole may be just as effective as 17beta-estradiol in suppressing bone loss due to ovariectomy but osthole perhaps does not work through the estrogen pathway. PMID- 12081140 TI - Metabolism of ginsenoside R(c) by human intestinal bacteria and its related antiallergic activity. AB - When ginsenoside R(c) was anaerobically incubated with human fecal microflora, all specimens metabolized ginsenoside R(c) to compound K and protopanaxadiol. The main metabolite was compound K. Among the bacteria isolated from human fecal microflora, most bacteria, such as Bacteroides sp., Eubacterium sp., and Bifidobacterium sp. potently transformed ginsenoside R(c) to compound K. Bifidobacterium K-103 and Eubacterium A-44 transformed it to compound K via ginsenoside R(d) and Bacteroides HJ-15 and Bifidobacterium K-506 metabolized to compound K via ginsenoside Mb, which was isolated as a new metabolite (M.W. 940[+Na]). Among ginsenoside R(c) and its metabolites, compound K exhibited the most potent antiallergic activity on the IgE-induced RBL cell line as well as potent cytotoxic activity against tumor cell lines. PMID- 12081141 TI - Study on the components of luobuma with peroxynitrite-scavenging activity. AB - The origin of the antioxidant activity of Luobuma aqueous extract was examined by measuring the peroxynitrite (ONOO-)-eliminating activities of fractions of this extract obtained by Sephadex LH-20 column chromatography. Three of the four fractions obtained, i.e., those excluding the H2O-eluted fraction, were found to possess (ONOO-)-eliminating activity. These three fractions were combined and fractionated again by Sephadex LH-20 column chromatography, which yielded five fractions. Seven different compounds were isolated from two of these five fractions with high activity. Epigallocatechin-(4beta-8)-epicatechin showed the highest (ONOO-)-eliminating activity. PMID- 12081142 TI - Antiproliferative activity of Vietnamese medicinal plants. AB - Methanol, methanol-water (1:1) and water extracts were prepared from seventy seven Vietnamese medicinal plants and tested for their antiproliferative activities against human HT-1080 fibrosarcoma cells. Among them, fifteen extracts including seven methanol extracts of Caesalpinia sappan, Catharanthus roseus, Coscinium fenestratum, Eurycoma longifolia, Hydnophytum formicarum and Streptocaulon juventas (collected at two areas), six methanol-water (1:1) extracts of Cae. sappan, Cat. roseus, Co. fenestratum, H. formicarum and S. juventas (at two areas), and two water extracts of Cae. sappan and S. juventas exhibited antiproliferative activities in a concentration-dependent manner. Their antiproliferative activities against human cervix HeLa adenocarcinoma, human lung A549 adenocarcinoma, murine colon 26-L5 carcinoma, murine Lewis lung carcinoma (LLC) and murine B16-BL6 melanoma cells were then examined. Co. fenestratum showed selective activity against lung carcinoma and/or lung metastatic cell lines, A549, LLC and B16-BL6, while H. formicarum and S. juventas showed selective activity against human tumor cell lines, HeLa and A549. Characteristic morphological change and DNA fragmentation indicated the antiproliferative activity to be due to the induction of apoptosis. PMID- 12081143 TI - Study on glutathionesulfonic acid sodium salt as biodistribution promoter for thiopental sodium. AB - The effects of glutathione (GSH) and glutathionesulfonic acid sodium salt [N-(N gamma-L-glutamyl-L-beta-sulfoalanyl)glycine sodium salt, GSO3Na], which is a minor metabolite of GSH, on the pharmacokinetics of thiopental sodium were investigated in rats. The concomitant use of GSO3Na with thiopental sodium significantly increased the tissue-to-plasma concentration ratio (Kp) of thiopental sodium 60 min after its administration in the heart, lung, brain, liver, kidney, and spleen, while GSH did not affect them. On the other hand, the Kp value of thiopental sodium 5 min after its administration with concomitant GSO3Na decreased significantly only in the spleen. Neither GSO3Na nor GSH changes the pharmacokinetic parameters of thiopental sodium. Significant change of the binding ratio of thiopental sodium to bovine serum albumin (BSA) was not observed by the addition of less than 5-fold GSO3Na. About 50% of thiopental sodium was bound to the brain, lung or liver, however, no significant change of this binding ratio was observed by the concomitant use of GSO3Na. The partition coefficient of thiopental sodium apparently increased by the concomitant use of GSO3Na but not by GSH. This phenomenon seemed to be concerned with a mechanism to increase the Kp values of thiopental sodium in the tissues. The increment in the drug distribution to tissues with concomitant GSO3Na observed in this study is useful information for the application of drug combinations as a biodistribution promoter. PMID- 12081144 TI - Liver targeting liposomes containing beta-sitosterol glucoside with regard to penetration-enhancing effect on HepG2 cells. AB - The aim of this study was to examine the interaction of soybean-derived sterylglucoside (SG) with the human hepatoblastoma cell line HepG2 with regard to the penetration-enhancing effect of beta-sitosterol glucoside (Sit-G) to clarify the accumulation of SG-containing liposomes (SG-liposomes) to the liver in vivo. The approach was based on measurement of the association of SG-liposomes labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (Dil) in terms of asialoglycoprotein receptor (ASGP-R)-mediated endocytosis, affinity of Sit-G using lAsys and the association of FITC-dextran 4400 (FD-4) increased by Sit-G with the cells. The association of SG-liposomes was decreased by addition of asialofetuin, suggesting that SG-liposomes might be taken up via ASGP-R. Sit-G showed higher affinity with HepG2 cells than HeLa cells, and enhanced the association of FD-4 depending on the incubation time and Sit-G concentrations. Significant positive correlations were found between Sit-G and FD-4 association with the cells, indicating that Sit-G enhanced the drug penetration by distribution in cell membranes. The high degree of liver association of SG liposomes in vivo might be related to recognition of glucose residues of SG by ASGP-R and to the high affinity and penetration-enhancing effect of Sit-G with hepatocytes. PMID- 12081145 TI - Cytotoxic effects of 27 anticancer drugs in HeLa and MDR1-overexpressing derivative cell lines. AB - The cytotoxic effects of 27 anticancer drugs including amrubicin, vinorelbine, paclitaxel, docetaxel, gemcitabine, and irinotecan were evaluated in human cervical carcinoma HeLa cells, and drug-resistant HeLa-derived Hvrl-1, HvrlO-6, and Hvr100-6 cells, which were newly established by stepwise exposure to vinblastine. FACS and RT-PCR analysis indicated that MDR1 (P-glycoprotein) was induced without any alterations in expression of its related transporters. Hvrl00 6 cells showed 2- to 200-fold higher resistance to anthracyclines than HeLa cells, and unexpectedly showed slight resistance to idarubicin and amrubicin. The relative resistance to vinca-alkaloids was 300- to 600,000-fold, and HvrlOO-6 cells showed the highest relative resistance to vinorelbine. HvrlOO-6 cells also showed 4000- and 60000-fold resistance to the taxanes paclitaxel and docetaxel, respectively. Hvr100-6 cells were also resistant to 6-mercaptopurine, actinomycin D, etoposide, and mitomycin C, with relative resistance of 8-, 45000-, 12-, and 9 fold, respectively. In contrast, HvrlOO-6 cells showed no or slight resistance to platinum derivatives, pyrimidine analogues, and alkylating agents or to irinotecan and its active form, or tamoxifen. The cytotoxicity of anthracyclines, vinca-alkaloids, taxanes, actinomycin D, and etoposide was extensively reversed by cyclosporin A. Cyclosporin A had no effect on the cytotoxicity of 6 mercaptopurine or mitomycin C, suggesting that resistance to these drugs was not mediated via MDR1. The alterations in cytotoxicity by overexpression of MDR1 and effects of cyclosporin A could be also qualitatively explained by [3H]vinblastine uptake experiments. The 27 anticancer drugs analyzed here could be classified into substrates and nonsubstrates for MDR1. This will be useful for designing effective regimens for chemotherapy. PMID- 12081146 TI - Transdermal delivery of indomethacin by iontophoresis. AB - The objective of this study was to construct a modified equation for the delivery of a drug by iontophoresis. Indomethacin was selected as a model since it has been widely used as a non-steroidal anti-inflammatory drug (NSAID) for external pharmaceutical preparations. The experiments were performed under a constant current in vivo using rat abdominal skin, and the plasma concentration was monitored by HPLC. Pharmacokinetic parameters were obtained from the plasma concentration profiles after intravenous injection. A theoretical value of the transdermal delivery of drug by iontophoresis was calculated from the plasma concentration and pharmacokinetic parameters. The experimental value was evidently higher than the theoretical one, suggesting the enhancement of passive diffusion with an increase of applied current. The modified equation was proposed for the delivery of a drug by iontophoresis incorporating enhanced passive diffusion. PMID- 12081147 TI - Potential usage of liposomal 4beta-aminoalkyl-4'-O-demethyl-4 desoxypodophyllotoxin (TOP-53) for cancer chemotherapy. AB - To enhance the therapeutic efficacy as well as to reduce the side effect, we attempted to liposomalize 4beta-aminoalkyl-4'-O-demethyl-4-desoxypodophyllotoxin (TOP-53), a novel and effective topoisomerase II inhibitor. More than 90% of TOP 53 was efficiently incorporated into the liposomes composed of dipalmitoylphosphatidylcholine and cholesterol by remote-loading method. Anti tumor activity of liposomal TOP-53 against solid tumor was examined in vivo using colon26 NL-17 carcinoma model mice. Three doses of liposomal TOP-53 (12 mg/kg/dose) showed significant tumor growth suppression (97.5% reduction determined at day 25) and the increase in life span (33%) of tumor-bearing mice. Furthermore, one mouse out of 5 was completely cured after treatment. Since similar efficacy was observed in the free TOP-53 treated group, liposomalization does not contribute much to the enhancement of therapeutic efficacy. However, a slight but measurable damage at the injection site was observed when free TOP-53 was injected, and the damage was diminished by the liposomalization. Taken together, liposomalization reduces the side effect rather than enhancing the therapeutic efficacy when TOP-53 is used. PMID- 12081148 TI - A novel oxidized low-density lipoprotein-binding protein, Asp-hemolysin, recognizes lysophosphatidylcholine. AB - Oxidized low-density lipoprotein (Ox-LDL) plays an important role in the initiation and progression of atherosclerosis. Asp-hemolysin, a hemolytic toxin from Aspergillus fumigatus, is a specific, high affinity binding protein for Ox LDL. We have previously shown that Ox-LDL strongly inhibits the hemolytic activity of Asp-hemolysin, and that the removal of lysophosphatidylcholine (lysoPC) from Ox-LDL abolished the inhibition. In the present study, to clarify the binding mechanism of Asp-hemolysin to Ox-LDL, we investigated the interaction between Asp-hemolysin and lysoPC as a typical lipid moiety of Ox-LDL. Based on western blot analysis, the binding of Asp-hemolysin to LDL, oxidized for different times, depended on the lysoPC content in each Ox-LDL. In addition, the inhibition of lysoPC production in Ox-LDL by phenylmethylsulfonyl fluoride (PMSF) pretreatment of LDL resulted in a marked decrease of Asp-hemolysin binding to PMSF-pretreated Ox-LDL. Furthermore, the binding analysis of Asp-hemolysin to lysoPC using ion-exchange chromatography revealed that Asp-hemolysin directly binds to lysoPC. PMID- 12081149 TI - Neuritogenesis of herbal (+)- and (-)-syringaresinols separated by chiral HPLC in PC12h and Neuro2a cells. AB - Syringaresinol isolated from Epimedium koreanum NAKA1 and Magnolia officinalis REHD. et WILS. was subjected to optical resolution by chiral HPLC to give (+)- and (-)-enantiomers. The two syringaresinol enantiomers, as well as a mixture of their glucosides, showed dose-dependent neuritogenesis in a concentration range from 0.24 to 24 microM in PC12h cells. PMID- 12081150 TI - Effects of ketoconazole on progesterone and cAMP production in MA-10 mouse Leydig tumor cells. AB - The effects of ketoconazole (KCZ) on secretion of progesterone and cAMP in Leydig cells were investigated in vitro. MA-10 mouse Leydig tumor cells were used to conduct the experiments. KCZ significantly inhibited the progesterone production from MA-10 cells in a dose dependent fashion between 2 and 20 microM among 1, 2 and 3 h of incubation. There was a statistically significant difference in luteinizing hormone (LH) stimulated progesterone production inhibited by 2 and 20 microm KCZ treatment compared to the control. The effect of KCZ on progesterone biosynthesis in MA-10 cells was mediated by cAMP, since KCZ suppressed basal and LH stimulated cAMP production and content within the same dose range. The stimulatory effects of forskolin and sodium fluoride on the adenylate cyclase system were also inhibited by KCZ. Moreover, dibutyryl cAMP blocked the inhibitory effect on steroidogenesis of KCZ in MA-10 cells. These data indicated that KCZ induced the inhibition of a catalytic component of adenylate cyclase holoenzyme in MA-10 mouse Leydig tumor cells. PMID- 12081151 TI - Synthesis and pharmacological activities of 1,8-naphthyridine derivatives. AB - In the present study, a series of 2-substituted-4-methyl-7-amino/4,7-dimethyl-1,8 naphthyridines were synthesized and characterized by IR, 1H-NMR and elemental analysis. The compounds were investigated for anticonvulsant (125, 250 mg/kg), cardiac and antimicrobial activities. The compounds were screened for antibacterial activity against gram (+) bacteria (Staphylococcus epidermidis, Bacillus subtilis, Enterococcusfaecalis and Micrococcus luteus) and gram (-) bacteria (Proteus vulgaris, Pseudomonas aeruginosa, Escherichia coli and Salmonella typhi). All the compounds except 2-(3'-phenylaminopropyloxy)-4-methyl 7-amino-1,8-naphthyridine exhibited significant anticonvulsant activity. The anticonvulsant activity of 2-(3-morpholino-2'-hydroxypropyloxy)-4-methyl-7-amino 1,8-naphthyridine, 2-(3'-diphenylamino-2'-hydroxypropyloxy)-4-methyl-7-amino-1,8 naphthyridine and 2-(3'-diethanolamino-propyloxy)-4,7-dimethy-1,8-naphthyridine at the dose of 250 mg/kg were found to be equivalent to diazepam (5 mg/kg). Sympathetic blocking activity was observed with 2-(3'-phenylamino-2' hydroxypropyloxy)-4-methyl-7-amino-1,8-naphthyridine, 2-(3'-diethanolamino-2' hydroxypropyloxy)-4-methyl-7-amino-1,8-naphthyridine and 2-(3'-diphenylamino-2' hydroxypropyloxy)-4-methyl-7-amino-1,8-naphthyridine only. All the compounds were devoid of antibacterial activity against the tested bacteria. PMID- 12081152 TI - Detection of oxidant-induced slight chromosomal damage in cells by subsequent exposure to X-rays. AB - We examined whether slight oxidative stress and/or damage in cells could be amplified by subsequent ionizing irradiation and thus become detectable as obvious chromosomal damage. WIL2-NS cells, a human B lymphoblastoid cell line, were pretreated with an oxidant and then exposed to X-rays at 0.25 or 0.5 Gy. The chromosomal damage in the cells was evaluated by cytokinesis-block micronucleus (CBMN) assay. Pretreatment with a superoxide-generating system (hypoxanthine (HX)/xanthine oxidase (XO), 1 and 2 mU/ml of XO), tert-butyl hydroperoxide (t BuOOH, 10 and 100 microM) or H2O2 (5 microM) alone did not induce significant chromosomal damage, but the oxidant-induced damage increased significantly with subsequent irradiation. The tested dose of these oxidants did not induce significant changes in cell viability, the nuclear division index, and the concentration of antioxidants, indicating that only weak oxidative stress was introduced into the cells. These results suggest that low-dose oxidant-induced chromosomal damage becomes detectable as obvious chromosomal damage with subsequent ionizing irradiation in vitro. PMID- 12081153 TI - Nobiletin as a tyrosinase inhibitor from the peel of Citrus fruit. AB - A tyrosinase inhibitor was isolated from the peel of Citrus fruit by activity guided fractionation, and identified as 3',4',5,6,7,8-hexamethoxyflavone (nobiletin) by comparison with reported spectral data. Nobiletin (IC50 of; 46.2 microM) exhibited more potency than Kojic acid (IC50; 77.4 microM) used as a positive control, and it was found to be potentially an effective inhibitor of the production of melanin. PMID- 12081154 TI - Anti-allergic effects of cnidii monnieri fructus (dried fruits of Cnidium monnieri) and its major component, osthol. AB - Anti-allergic effects (types I and IV) of the 70% ethanol extract (CM-ext) obtained from Cnidii Monnieri Fructus (dried fruits of Cnidium monnieri) were investigated on 48 h homologous passive cutaneous anaphylaxis (PCA), 2, 4 dinitrofluorobenzene (DNFB)-induced contact dermatitis and picryl chloride (PC) induced contact dermatitis in experimental animals. CM-ext showed inhibitory effects on these allergic models. Osthol isolated from CM-ext also had the inhibitory effects. These results suggested that Cnidii Monnieri Fructus might be useful as an agent for allergic diseases and that its anti-allergic effect was partially attributable to a coumarin derivative, osthol. PMID- 12081155 TI - Pharmacokinetics of propofol in elderly coronary artery bypass graft patients under total intravenous anesthesia. AB - The present paper investigates the pharmacokinetics of propofol in the plasma of two elderly patients operated on under total intravenous anesthesia using propofol. A 78-year-old (patient A) and a 76-year-old (patient B), both Japanese men with unstable angina pectoris, were operated on for coronary artery bypass grafts. For the induction of anesthesia, 1.5 mg/kg propofol was administered as a single bolus infusion, and anesthesia was maintained using the step-down infusion regimens of propofol. Propofol concentration in the plasma was measured by HPLC with a fluorescence detector. The simulation curves, following the two compartment model, fitted well to the profiles of the individual data of propofol concentrations in the plasma. When 4 mg/kg/h of propofol was administered to both patients while maintaining anesthesia, propofol concentrations in the plasma were maintained at over 1.0 microg/ml. In patient A, the propofol concentration in the plasma was 140 ng/ml at 6 h after the end of the infusion. In patient B, the propofol concentrations in the plasma were 73 ng/ml at 6 h and 35 ng/ml at 12 h after the end of the infusion. The apparent distribution volumes of patients A and B were 1.43 and 1.62 l/kg, respectively. The half-lives of propofol in the plasma of patients A and B were estimated to be 13.3 and 17.4 min as the a phase, and 10.1 and 10.5 h as the beta phase, respectively. In elderly patients with cardiac surgery, the maintenance concentrations of propofol in the plasma were enough to maintain a concentration of 1.0 microg/ml, and the half-life may be longer than previously reported values in adult patients. PMID- 12081156 TI - Systemic inflammatory response associated with augmentation and activation of leukocytes in Candida/indomethacin administered mice. AB - We have previously shown that the combination of beta-glucan and nonsteroidal anti-inflammatory drugs (NSAIDs) was lethal to mice. In this study, we examined the effect of Candida on this treatment and found that Candida showed similar lethal toxicity when used in combination with indomethacin. It was also confirmed that LPS preparations from various origins and by various procedures did not show lethality. Candida/indomethacin induced lethality was found to be associated with increased number of leukocytes in various organs and activation of these cells. These findings strongly suggest that pathogenic fungi augment the side effects of non-steroidal anti-inflammatory drugs. PMID- 12081157 TI - Histamine and cutaneous allergy: old friend, new player. PMID- 12081158 TI - Dapsone in Behcet's disease: a double-blind, placebo-controlled, cross-over study. AB - The study was designed to investigate the effects of dapsone in the treatment of mucocutaneous manifestations of Behcet's disease and the possible prophylactic role of dapsone in a double blind/placebo controlled clinical trial. Twenty patients diagnosed according to the International Study Group criteria as Behcet's disease were included in the study. Patients were randomly allocated to receive either dapsone 100 mg daily or placebo for three months in a double-blind manner. After three months, patients were crossed over and followed for a further three months. Patients were followed up in each visit by assessing the number, size, duration and frequency of oral and genital ulcers, other cutaneous manifestations, and systemic manifestations of the disease. A pathergy test was done on each visit. Laboratory investigations included hemoglobin concentration, white blood cell count, ESR, and C-reactive protein. In dapsone-treated patients, there were significant reductions in the oral and genital ulcer parameters as well as the incidence of other cutanous and systemic manifestations. In the placebo-treated group, there were no significant changes in these parameters. The pathergy test result as well as those of other laboratory tests were all decreased in the dapsone-treated group. Although this study was a small scale study, it shows that dapsone was effective in treatment of mucocutaneous manifestations of Behcet's disease and possibly in prophylaxis against systemic manifestations of the disease. This result should lead to a larger scale study with a longer duration of follow-up. PMID- 12081159 TI - Activation of mast cells within a tumor of angiosarcoma: ultrastructural study of five cases. AB - The accumulation of mast cells around tumors is a well-recognized phenomenon in a number of malignancies, including basal cell carcinoma, melanoma, and breast cancer. However, little information exists regarding mast cells within tumor nests. To clarify the role of mast cells infiltrating in skin cancers, we examined the morphological features of mast cells within tumors of five cases of angiosarcoma, including two patients with Stewart-Treves syndrome, by electron microscopy. In light microscopy, mast cells were observed within tumor nests at various densities and exhibited weak staining intensity with toluidine blue. By electron microscopy, most of the tumor-infiltrating mast cells exhibited anaphylactic or piecemeal degranulation, indicating that the mast cells had been activated in situ. Some mast cells were noted in close apposition to tumor cells, suggesting the existence of direct cell-to-cell interactions. Tumor cells adjacent to mast cells showed no degenerative changes. In conclusion, these results suggest that careful histologic examination in combination with electron microscopy should enable us to identify more mast cells within cancer lesions with greater sensitivity than in a number of prior reports. Furthermore, the close proximity of mast cells and surrounding tumor cells suggests some biologically significant role of mast cells in the development of angiosarcoma, including tumor growth as well as host immunity and stromal reaction. PMID- 12081160 TI - Report of eight infants with acute infantile hemorrhagic edema and review of the literature. AB - Acute infantile hemorrhagic edema (AIHE) is a cutaneous leukocytoclastic vasculitis, clinically characterized by the symptom triad of fever, large purpuric skin lesions, and edema. The clinical picture has a violent onset, a short benign course, and spontaneous complete recovery. In this article, we present eight patients who were admitted with rashes on the skin and edema on the eyelids and extremities, and were diagnosed with AIHE according to their clinical and histopathological features (immunohistological study was also performed in three of them). Our purpose was to emphasize that, aside from Henoch-Schonlein purpura, meningococcemia, septicemia, and purpura fulminans, AIHE benign disorder should also be considered in the differential diagnosis to determine the clinical course and treatment protocol in patients with purpuric rashes. PMID- 12081161 TI - Multiple cutaneous metastases from carcinoma of the penis. AB - A 65-year-old male patient with carcinoma of the penis in association with bilateral inguinal lymphadenopathy and widespread cutaneous dissemination is described. Histopathological examination of the primary tumor and metastatic sites was suggestive of undifferentiated squamous cell carcinoma. The patient was administered combination chemothrapy using cisplatin, methotrexate, and bleomycin with a good response after three cycles. The case is unique because of the rapid onset and widespread nature of skin metastasis and rare presentation. PMID- 12081162 TI - A case of cutaneous ganglioneuroma developing within a lesion of seborrheic keratosis. AB - A 67-year-old man noticed on the abdominal skin a coin-sized verrucous plaque, in which a small granulation-like lesion developed several weeks before the first consultation at our clinic. The histology of the biopsied granulation-like lesion showed a well demarcated, but not associated with the collagenous capsule, mass of proliferating eosinophilic cells, consisting of intervening spindle-shaped Schwann cells and mature ganglion cells scattered in the tumor nest. We diagnosed this tumor nest as a ganglioneuroma, which rarely develops in the skin tissue. The simply resected surrounding verrucous lesion was histologically seborrheic keratosis. PMID- 12081163 TI - Co-existence of variants of porokeratosis: a case report and a review of the literature. AB - Rarely, different variants of porokeratosis may coexist in an individual patient or their family members. A patient with the linear form of porokeratosis present since birth subsequently developed the disseminated superficial actinic form at a later age. A review of the literature pertaining to the coexistence of variants of porokeratosis suggests a significant association between the linear and disseminated superficial actinic forms. Genetic linkage between different variants and the basis for their association is discussed. PMID- 12081164 TI - Intradermal nodular fasciitis of the face. AB - Nodular fasciitis represents reactive myofibroblastic proliferative lesions in the subcutaneous tissue, skeletal muscles, and fascia. Uncommon clinical and pathological variants of nodular fasciitis, such as intravascular, cranial, ossifying, and proliferative fasciitis, have been described. Six cases documenting a dermal location of nodular fasciitis have been reported in the English literature. Some authors have described that the histopathological features of intradermal nodular fasciitis are virtually identical to those of postoperative/posttraumatic spindle cell nodule of the skin. We herein report a 41-year-old Korean woman presenting with a 0.5 cm facial papule in which nodular fasciitis occurred in the dermis. PMID- 12081165 TI - Chromoblastomycosis: report of two cases from Nepal. AB - Chromoblastomycosis is a chronic fungal infection of the skin and subcutaneous tissue caused by dematiaceous fungi. The first case, a 67-year-old male farmer, presented with itchy hyperkeratotic, scaly plaques with scarring and black dots on the lateral aspects of his left arm and dorsum of his left hand of 28 years duration. The case was clinically diagnosed as chromoblastomycosis. The second case, a 75-year-old farmer, presented with erythematous, crusted, scaly plaques on the dorsum of the left foot of 30 years duration. Initially, a clinical diagnosis of lupus vulgaris was made, but treatment with anti tuberculosis therapy showed no improvement. On the basis of histopathological examinations of skin biopsies and isolation of fungus on culture, both cases were diagnosed as chromoblastomycosis. To the best of our knowledge, these two cases are the first case reports of chromoblastomycosis from Nepal and are presented for their academic interest. PMID- 12081166 TI - A case of hyper-IgE syndrome complicated by cutaneous, nodal, and liver peripheral T cell lymphomas. PMID- 12081167 TI - Cutaneous malignant hemangiopericytoma with unusual clinical behavior. PMID- 12081168 TI - Outreach health adviser in a community clinic screening programme improves management of genital chlamydia infection. AB - OBJECTIVE: To assess the effectiveness of an outreach health adviser on treatment, partner notification and outcome for clients diagnosed with genital chlamydia (CT) infection at a community young people's clinic. METHODS: From August 1999 to March 2000, a genitourinary medicine (GUM) based health adviser helped to develop testing and undertook outreach management of clients aged under 26 years diagnosed with CT infection. In addition to facilitating referral to GUM, she gave antibiotic treatment based on a GUM derived patient group direction to those not wishing to travel to the GUM clinic. She also advised them on contact tracing and the need for a compliance check (CC). RESULTS: Chlamydia positive tests with ligase chain reaction (LCR), on first void urine, were obtained for 62 (12.9%) of 481 female clients, one (5%) of 20 male clients, and nine (53%) of 17 male contacts of female positive cases. All 72 testing positive received their result and were treated. Two urine samples positive for CT showed positive LCR tests for gonorrhoea. Proportions of named contacts seen (67%) and reattendances for compliance checks (60%) were similar to those for women seen in GUM services. CONCLUSIONS: Health adviser input with the ability to treat can be effective in reducing the growth of identified but untreated genital chlamydia infection consequent upon community based screening. Such a strategy appears comparable with, and can add to, GUM based treatment of infection. It helps to address the need for alternative management strategies in the light of the national sexual health strategy. PMID- 12081169 TI - Gonorrhoea in London: usefulness of first line therapies. AB - OBJECTIVES: To determine the true prevalence and patterns of resistance of Neisseria gonorrhoeae to antimicrobial agents used for therapy in London. METHODS: A longitudinal study of a representative sample of isolates of N gonorrhoeae linked to demographic data of infected patients was undertaken. Isolates were collected from consecutive patients attending genitourinary medicine clinics in the North and South Thames regions of London during 3 months of each of 4 years, 1997-2000. Prevalence of plasmid mediated resistance to penicillin and chromosomally mediated resistance to penicillin and ciprofloxacin was determined by susceptibility testing. The association of antimicrobial resistance among gonococci with sexual orientation of the patient and country of acquisition of infection was determined. RESULTS: Numbers of gonococcal isolates collected over the same time period between 1997-2000 have increased by 74%. Plasmid mediated resistance to penicillin was low but has risen above 5% in 2000. Chromosomal resistance to penicillin has fallen below the 5% level but total resistance to penicillin, plasmid and chromosomally mediated, was above 5% in all 4 years. The incidence of resistance and reduced susceptibility to the alternative therapeutic choice, ciprofloxacin, is low but numbers are increasing in each year. High level resistance, to both penicillin and ciprofloxacin, has generally been found among heterosexual men and is often acquired abroad. However, there is some evidence of a change to endemic spread in 2000. CONCLUSIONS: This surveillance programme shows that the epidemiology of gonorrhoea in London is changing with increasing numbers and changing patterns of resistance. If gonorrhoea is to be controlled and targets set by the sexual health strategy are to be met, intervention with effective and appropriate antimicrobial agents is essential. PMID- 12081170 TI - Decreasing STD incidence and increasing condom use among Chinese sex workers following a short term intervention: a prospective cohort study. AB - OBJECTIVE: To describe the impact of a repeating behavioural intervention focused on preventive education and provision of STD testing and treatment services to female sex workers. METHODS: A prospective cohort study of 966 sex workers (first of its kind in China) was conducted in Guangzhou from March 1998 to October 1999. At each visit information was collected on sexual behaviour, condom use and knowledge about HIV transmission and condom use, education was given, STD were diagnosed, and treatment was provided free of charge. We evaluated trends in condom use, knowledge about HIV transmission and condom use, and STD incidences. Generalised estimating equations were applied to control for repeated measurements. RESULTS: The proportion of consistent condom use increased from the intake through the third follow up visit (from 30% to 81%), as well as the proportion of having good knowledge on HIV transmission (4.3% to 98.6%) and condom use (23.6% to 79.3%). The incidence of gonorrhoea, trichomoniasis, and chlamydia decreased over each follow up visit; from 17.5/100 person years (PY), 22.4/100PY, 65.9/100PY at the first follow up visit to 5.1/100PY, 3.0/100PY, 16.1/100PY at the third follow up visit, for each STD respectively. CONCLUSIONS: STD care and prevention programmes targeting sex workers are feasible in China and, more importantly, intervention consisting of prevention education and STD care is likely to reduce unprotected sexual behaviour and STD incidence in this group at high risk for HIV and other STD infection. PMID- 12081171 TI - Chlamydia trachomatis infections in eastern Europe: legal aspects, epidemiology, diagnosis, and treatment. AB - OBJECTIVES: Knowledge concerning genital Chlamydia trachomatis infections in eastern Europe is scarce. Data on the legal aspects, epidemiology, diagnosis, and treatment of the infection have never been collected, summarised, and presented to the international scientific community. The aim of this study was to present the current situation on the main aspects of chlamydial infections in the countries of eastern Europe. METHODS: Written questionnaires concerning legal aspects, epidemiology, diagnosis, and treatment of the infection were distributed among national STI operating administrators as well as researchers who had presented papers at earlier meetings of European chlamydia or STI societies. RESULTS: Most of the countries have not legalised reporting of chlamydial infections and in those who have done so, the quality of the reporting system is poor. Contact tracing is mostly done on a voluntary basis. Reported chlamydia incidence varies from 21 to 276 per 100000 inhabitants. The most commonly used diagnostic test remains the direct immunofluorescence test; however, some tendencies towards nucleic acid amplification are in evidence. Diagnostic services are paid for by the patient himself, while treatment in many countries is partially or completely covered by public insurance. CONCLUSIONS: This is the first report summarising data concerning the situation on C trachomatis infections in eastern Europe. The reporting system and diagnosis of C trachomatis infections remain suboptimal, which allows neither control of the epidemiological situation nor optimal treatment of the patients. The most urgent work currently necessary is the education of professionals and the general population. PMID- 12081172 TI - Sexually transmitted infections in Bangladeshis resident in the UK: a case control study. AB - OBJECTIVE: To evaluate the prevalence of sexually transmitted infections (STIs) and mode of presentation in patients originating from Bangladesh and resident in the United Kingdom in comparison with non-Bangladeshi patients attending an inner London genitourinary medicine (GUM) clinic. METHODS: A retrospective, cross sectional study with comparator group was carried out at an open access GUM clinic in east London. 104 consecutive newly attending Bangladeshi men were compared with 199 consecutive newly attending non-Bangladeshi men and 115 consecutive newly attending Bangladeshi women were compared with 218 consecutive newly attending non-Bangladeshi women. Any diagnosed sexually transmitted infections, sexual history characteristics, reasons for presentation, and referral patterns were noted. RESULTS: Bangladeshi men (28.8% compared with 7.5%; p<0.0001) and women (42.7% compared with 12.8%; p<0.0001) were more likely to be referred by their general practitioners or other medical services. Bangladeshi men were more likely to present with sexual dysfunction (12.5% compared with 2.5%; p=0.001). The prevalence of STIs was broadly similar across the study groups; however, syphilis was significantly more common in the Bangladeshi men (10.9% compared with 4%; p=0.04) and nongonococcal urethritis (NGU) in the control men (35% compared with 20.2%; p=0.02). Bacterial vaginosis was an infrequent diagnosis in the Bangladeshi women (3.5% compared with 22.4%; p<0.0001). CONCLUSIONS: STI prevalence in Bangladeshis attending GUM services is similar to other populations although patterns of presentation and referral do show variation. Bangladeshi men are more likely to access GUM clinics for psychosexual services. The presence of STIs in Bangladeshis particularly those imported from Bangladesh provides an opportunity for HIV transmission between the United Kingdom and Bangladesh. PMID- 12081173 TI - Low incidence of syphilis among factory workers in Ethiopia: effect of an intervention based on education and counselling. AB - BACKGROUND: The prevalence and incidence of syphilis infection were examined in a cohort study of factory workers in Ethiopia. METHOD: Between February 1997 and March 1999, 409 men and 348 women were enrolled and followed in the cohort study. RESULTS: The prevalence (95% CI) of past/current syphilis (positive TPPA serology) was 28.9% (25.7% to 32.3%), and factors associated with past/current syphilis were markers of risky sexual behaviours including HIV infection. In this cohort of factory workers subject to public information/education meetings, testing for HIV antibodies, and individual counselling, the incidence (97.5% one sided CI) of new syphilis infections was 0/691 = 0 (0 to 0.5) per 100 person years. CONCLUSION: This study has documented a reduction in risky sexual behaviours and a low syphilis incidence among factory workers participating in a cohort study on HIV infection progression in Addis Ababa. PMID- 12081174 TI - Health seeking and sexual behaviour in patients with sexually transmitted infections: the importance of traditional healers in Thyolo, Malawi. AB - OBJECTIVES: To describe health seeking and sexual behaviour including condom use among patients presenting with sexually transmitted infections (STIs) and, to identify sociodemographic and behavioural risk factors associated with "no condom use" during the symptomatic period. METHODS: A cross sectional study of consecutive new STI cases presenting at the district STI clinic in Thyolo, Malawi. They were interviewed by STI counsellors after obtaining informed consent. All patients were treated according to national guidelines. RESULTS: Out of 498 new STI clients, 53% had taken some form of medication before coming to the STI clinic, the most frequent alternative source being the traditional healer (37%). 46% of all clients reported sex during the symptomatic period (median 14 days), the majority (74%) not using condoms. 90% of all those who had not used condoms resided in villages and had seen only the traditional healer. Significant risk factors associated with "no condom use" included visiting a traditional healer, being female, having less than 8 years of school education, and being resident in villages. Genital ulcer disease (GUD) was the most common STI in males (49%) while in females this comprised 27% of STIs. CONCLUSIONS: These findings, and especially the extremely high GUD prevalence is of particular concern, considering the high national HIV prevalence in Malawi (9%) and the implications for STI and HIV transmission. There is an urgent need to integrate traditional healers in control activities, encourage their role in promoting safer sexual behaviour, and to reorient or even change existing strategies on condom promotion and STI control. PMID- 12081175 TI - Post-exposure prophylaxis for non-occupational exposure to HIV: current clinical practice and opinions in the UK. AB - OBJECTIVES: To assess the frequency and nature of requests for post-exposure prophylaxis following nonoccupational exposure (NONOPEP) to HIV and to describe variations in practice and opinions on the need for its administration at UK genitourinary medicine (GUM) clinics. METHOD: A retrospective survey was carried out of physicians representative of all UK GUM clinics using self completed questionnaires requesting information for January to December 1999. The number of requests for NONOPEP, reasons for the requests, the number prescribed, and physician opinions regarding the justification for its administration were noted. RESULTS: The number of requests and prescriptions for NONOPEP increased fourfold and sevenfold respectively in comparison with a survey from 1997. Of 242 requests, 130 people were prescribed NONOPEP. Half the requests followed sexual exposures between known HIV discordant couples. Requests for NONOPEP were received by 56 of 132 (42%) clinics, with nine clinics receiving over half of them (145/242, 60%). Similarly, over half the prescriptions for NONOPEP (83/130, 64%) were given by six of 39 prescribing clinics. Most physicians thought that post-exposure prophylaxis (PEP) was justified for people exposed to a known HIV positive source patient resulting from sexual assault or unprotected receptive anal or penovaginal sex. CONCLUSION: The use of NONOPEP has increased since the last survey and there is considerable variation between GUM clinics in practice and beliefs regarding administration of NONOPEP. PMID- 12081176 TI - Offering routine antenatal testing for HIV and hepatitis B in the rural setting of Cornwall. AB - Before Department of Health directives to offer HIV and hepatitis B (HBV) testing to all pregnant women, there was little such screening in Cornwall. Through a multiagency collaborative approach a new antenatal screening programme for HIV/HBV has been introduced with high uptake in the first year (HIV 92% and HBV 93%). We also report the findings of a questionnaire survey of community midwives who alone offer antenatal HIV/HBV testing in Cornwall. PMID- 12081178 TI - Lactic acidosis and abnormal liver function in advanced HIV disease. AB - A 48 year old man receiving HAART presented with late stage HIV disease, non specific symptoms, a normal sized liver, ascites, and lactic acidosis. Following a failed liver biopsy worsening acidosis developed, requiring ICU support. Progressive liver failure occurred. Endoscopy showed oesophageal varices and a transjugular liver biopsy showed non-cirrhotic cholestasis; findings that were ascribed to HAART. PMID- 12081177 TI - Contamination of environmental surfaces by genital human papillomaviruses. AB - OBJECTIVE: To investigate contamination of environmental surfaces with human papillomaviruses (HPV) DNA in two genitourinary medicine (GUM) clinics and in an on-site staff leisure and fitness centre. METHODS: Samples were collected from the treatment rooms and patients' toilets of two GUM clinics situated at two hospital sites and from the toilets of the staff leisure and fitness centre on one of the sites. Samples were tested for the presence of HPV DNA by nested polymerase chain reaction (PCR), and HPV amplicons were typed by reverse line hybridisation using HPV type specific oligonucleotide probes complementary to 35 HPV types. All samples were also tested for beta globin DNA by PCR in order to assess their quality. RESULTS: HPV DNA was found to be present at more than 50% of the sites sampled in one of the GUM clinics, but was absent in the second, and also from the staff leisure and fitness centre. All HPV DNA detected was found to be cell associated. The most commonly found HPV types were 6, 11, and 16, respectively. HPV infected cells were found to be localised mainly to surfaces used predominantly by medical staff. CONCLUSIONS: This study has identified contamination of the environment of a GUM clinic. Possible sources for the contamination of the clinic toilets were from genital sites via hands to the environment. Within the treatment rooms the most likely route of HPV DNA contamination of the environment was via the doctor's gloved hands. PMID- 12081179 TI - Genital ulcers, stigma, HIV, and STI control in sub-Saharan Africa. AB - HIV associated stigma is still prevalent throughout Africa despite the spread of the epidemic. Stigma is also attached to sexually transmitted infections (STIs). Despite the importance of STIs, particularly genital ulceration in facilitating heterosexual HIV transmission, policymakers continue to focus mainly on other priorities. It would appear that this lack of public health initiative in tackling genital ulcers is itself an example of stigmatisation. Possible explanations for this include geographical variation in the data and a perception that genital ulcers are not a topic that can be discussed freely and openly. HIV policymakers in countries worst affected by HIV in Africa should examine their own opinions for bias when determining public health priorities for HIV prevention. The importance of genital ulcers should be reassessed and publicised. PMID- 12081180 TI - Prostatic tuberculosis in an HIV infected male. PMID- 12081181 TI - 8th European Conference on Clinical Aspects and Treatment of HIV Infection, Athens, 28-31 October 2001. PMID- 12081182 TI - Report on the MSSVD Spring meeting, Belfast, 24-26 May 2001. PMID- 12081185 TI - Vaginitis emphysematosa. PMID- 12081186 TI - Management of screened chlamydia positive women. PMID- 12081187 TI - Short term interventions are not enough. PMID- 12081188 TI - Revised UK national guidelines on sexually transmitted infections and closely related conditions 2002. PMID- 12081189 TI - The national strategy for sexual health and HIV: implications for genitourinary medicine. AB - The first ever national strategy for sexual health and HIV in England was published in July 2001 and proposes a comprehensive and holistic model for dealing with an increasing public health problem. The strategy covers the issues of prevention, service provision, commissioning of services, and the necessary requirements to support change. This paper concentrates on service issues and developments in relation to genitourinary medicine/HIV services, and outlines a model for providing these which attempts to do so around patients' needs, delivered through comprehensive and interlinked local networks of services. PMID- 12081190 TI - The application of qualitative research methods to the study of sexually transmitted infections. AB - Qualitative methods have gained support in health sciences research and have a role to play in STI studies. This article reviews the main techniques used by qualitative researchers, with illustrations taken from the field of STIs. PMID- 12081191 TI - Lymphogranuloma venereum. AB - Lymphogranuloma venereum is caused by serovars of Chlamydia trachomatis. It is rare in industrialised countries, but is endemic in parts of Africa, Asia, South America, and the Caribbean. We discuss its clinical features, diagnosis, and management. PMID- 12081192 TI - Sexual health clinics for women led by specialist nurses or senior house officers in a central London GUM service: a randomised controlled trial. AB - OBJECTIVES: To assess the care process and clinical outcomes for two different models of GUM clinic for women: one led by specialist nurses and the other by senior house officers (SHOs). METHOD: An open randomised controlled trial was carried out in a central London genitourinary medicine (GUM) women's clinic. Of 1172 women telephoning for an appointment, 880 were randomised to provide 169 eligible patients in the specialist nurse arm and 178 in the SHO arm. Of the eligible patients a total of 224 attended their appointment. The clinical records of the randomised women were audited for adequacy of care according to local guidelines. 30 key variables were objectively assessed and recorded on a standard audit form. An overall unitary index score (%) was calculated for each patient. The main variables associated with the outcome of specialist nurse and SHO decision making (diagnostic test request, preliminary diagnosis, and treatment provided) were then analysed independently. RESULTS: The median documentation audit scores for specialist nurses (n=103) and SHOs (n=121) were 92% and 85% respectively (p<0.0001). The specialist nurses' documentation was significantly (p<0.05) more complete than the SHOs' for five variables: details of menstrual cycle, physical examination, medication instructions given to patients, health promotion discussion, and provision of condoms. Specialist nurses performed equally to the SHOs with regard to requesting the correct diagnostic tests, providing the correct preliminary diagnosis, and providing the correct treatment. CONCLUSIONS: A model of care using trained GUM nurses working within agreed protocols can provide comprehensive patient care for female patients that is equal to care provided by SHOs. Our results raise important issues regarding advanced GUM nursing education and training, protocol development, and accountability. PMID- 12081193 TI - A postal survey to identify and describe nurse led clinics in genitourinary medicine services across England. AB - BACKGROUND: Nurses in genitourinary medicine (GUM) services are progressively extending their roles to conduct "comprehensive care" nurse led clinics. In such roles the nurse coordinates the first line, comprehensive care of patients presenting with sexual health conditions and issues. OBJECTIVES: To identify and describe comprehensive care nurse led clinics in GUM services across England. METHODS: A postal questionnaire consisting of 17 closed response questions was sent to 209 GUM services across England. A second questionnaire was sent to non responders to increase the response rate. Data were single entered and analysed using SPSS. RESULTS: Of the 190 GUM clinic respondents (91% response rate), 44 (23%) reported providing some form of comprehensive care nurse led clinic, 90% of which were initiated since 1995. Key results show staff development featured as the main reason for initiating such services and there was general consistency in the aspects of care undertaken by these nurses. There was evidence of guideline development specific to nurse led care and some patient group direction use for supplying medication. The level of support from medical staff while nurse led clinics were being conducted varied between services. Few services have conducted any audit or research to monitor/evaluate nurse led care. There was little consistency in the clinical experience and educational prerequisites to undertake comprehensive care nurse led clinics. Continuing professional development opportunities also varied between services. CONCLUSIONS: The steady growth of comprehensive care nurse led clinics indicates that the skills of GUM nurses are being recognised. Nurses working in advanced practice roles now require courses and study days reflecting these changes in practice. Locally agreed practice guidelines can define nursing practice boundaries and ensure accountability, as will the development of patient group directions to supply medication. Monitoring and evaluation of nurse led clinics also require attention. PMID- 12081194 TI - The glomerulo-tubular junction: a target in renal diseases. AB - Both global and segmental glomerulopathies may damage specific areas of the renal glomerulus. Diseases associated with glomerular hyperperfusion cause lesions at the vascular pole, while diseases associated with proteinuria often damage the tubular pole. Atubular glomeruli are now known to be plentiful in a variety of common renal diseases. These glomeruli are disconnected from their tubule at the tubular pole and therefore cannot participate in the production of urine. It is widely believed that the disconnection is a result of external compression by periglomerular fibrosis. However, the variable anatomy and cell populations within both the glomerulus and the beginning of the proximal tubule at the glomerulo-tubular junction may also have important roles to play in the response to damage at this sensitive site of the nephron. PMID- 12081195 TI - Expression of the duffy antigen/receptor for chemokines (DARC) by the inflamed synovial endothelium. AB - The expression of chemokine binding sites on the endothelial cells of venules in inflamed synovia was examined and whether the Duffy antigen/receptor for chemokines (DARC) was involved. In situ binding assays were performed to determine the expression of chemokine binding sites from rheumatoid (n = 10) and non-rheumatoid (n = 10) synovia. The expression of DARC protein and mRNA was examined by immunohistochemistry and northern blotting. The involvement of DARC in chemokine binding was studied by incubating sections with blocking antibodies to DARC (Fy3 and 6), to find out if these reduced 125I-IL-8 binding. Binding of radiolabelled chemokines IL-8, RANTES, MCP-1, but not MIP-1alpha, was found on venular endothelial cells in inflamed synovia from both rheumatoid and non rheumatoid patients. Excess homologous unlabelled chemokine displaced binding and excess unlabelled RANTES could displace radiolabelled IL-8 binding. DARC protein expression was demonstrated on venular endothelial cells in all samples and DARC mRNA could be detected in extracts from synovia. There was downregulation of DARC protein and mRNA in rheumatoid samples. Binding of IL-8 to both rheumatoid and non-rheumatoid synovia was significantly reduced in the presence of anti-DARC Fy3 and Fy6 monoclonal antibodies. These findings show the expression of a multispecific chemokine binding site on the inflamed synovial endothelium, with evidence for involvement of DARC. This suggests a potential role for DARC in the inflammatory processes involved in synovitis. PMID- 12081196 TI - Skin flap-induced regression of granulation tissue correlates with reduced growth factor and increased metalloproteinase expression. AB - Previous studies have shown that covering granulation tissue of a full-thickness skin wound by a vascularized skin flap induces tissue remodeling, with a rapid loss of granulation tissue cells by apoptosis. In the present study, in situ hybridization has been used to examine mRNA expression for several factors that may be implicated in the apoptosis seen in this tissue. Skin wounds were made on the dorsal skin of 8-week-old rats. Ten days after wounding, skin flaps were created surgically and sutured over the granulation tissue. Tissue sections of granulation tissue from various times after addition of the skin flap were hybridized with 33P-labelled cRNA probes for transforming growth factor-beta1 (TGF-beta1), beta-inducible gene H3 (beta-ig-h3), alpha1 (1) procollagen, alpha smooth muscle actin, matrix metalloproteinase-13 (MMP-13) and -2 (MMP-2), tissue inhibitor of metalloproteinase-1 (TIMP-1), and inducible nitric oxide synthase (iNOS). Control granulation tissue prior to addition of the skin flap showed high levels of TGF-beta1, beta-ig-h3, alpha1 (1) procollagen, alpha-smooth muscle actin, and TIMP-1 expression. MMP-13, MMP-2, and iNOS mRNA were low in 10-day granulation tissue. Addition of a skin flap resulted in a decrease in the expression of TGF-beta1, beta-ig-h3, alpha1 (I) procollagen, alpha-smooth muscle actin, and TIMP-1, but increased expression of MMP-13 and MMP-2. Similarly, an increase in iNOS mRNA expression was observed in the granulation tissue after addition of the skin flap. Addition of a vascularized skin flap may result in rapid remodelling of granulation tissue due to a decrease in expression of the trophic growth factor TGF-beta1 and increased degradation of extracellular matrix due to an alteration in the balance between MMPs and their inhibitor, TIMP-1. Additionally, increased iNOS expression may also favour apoptosis through the generation of free radicals. The additive effect of reduced growth factor expression, increased extracellular matrix turnover, and nitric oxide generation may result in the fibroblast and vascular cell apoptosis seen during the rapid remodelling of this tissue. PMID- 12081197 TI - Cyclin D1 overexpression in colorectal carcinoma in vivo is dependent on beta catenin protein dysregulation, but not k-ras mutation. AB - Cyclin D1 protein overexpression is commonly found in colorectal carcinomas (CRCs) and is associated with a poorer prognosis, but the mechanism underlying overexpression remains uncertain. Both dysregulation of beta-catenin protein expression and k-ras mutation have recently been shown to promote cyclin D1 expression in human in vitro and rodent in vivo studies. In this study, 53 sporadic CRCs were examined by immunohistochemistry for cyclin D1 and beta catenin protein expression, and with PCR and direct DNA sequencing for k-ras gene status. The study also addressed whether cyclin Dl overexpression might associate with poorer prognosis because of a relationship with poorer response to 5 fluorouracil (5FU) chemotherapy. Cyclin D1 overexpression was demonstrated in 34/53 (64%) CRCs, was significantly associated with higher Dukes' stage, and was particularly prominent at the invasive edges of carcinomas. Furthermore, cyclin D1 overexpression was always and only seen in association with nuclear expression of beta-catenin. There were no significant associations between cyclin D1 overexpression and k-ras mutation or response to 5FU. Amongst 17 microsatellite unstable CRCs, a smaller proportion of tumours showed cyclin D1 overexpression (18%), but again cyclin D1 overexpression was only seen in cases showing nuclear beta-catenin expression. In conclusion, beta-catenin protein dysregulation, but not k-ras mutation, appears to be required for cyclin D1 overexpression in colorectal carcinoma in vivo. PMID- 12081198 TI - Glomerulo-tubular junction stenosis as a factor contributing to glomerular obsolescence in IgA nephropathy. AB - Periglomerular fibrosis (PF) is an interstitial injury observed in various renal diseases. It is speculated that this lesion, by occluding the glomerulo-tubular junction (GTJ) and causing atubular glomeruli, may result functionally in a reduction of the glomerular filtration rate (GFR) and may be a factor contributing to the progression of renal disease. In the present study, 340 renal biopsy specimens were analysed to determine whether or not there was nephron injury derived from such a mechanism, as well as direct glomerular injury, in IgA nephropathy (IgAN). The patients were divided into five groups according to the degree of glomerular sclerosis. The average age was lower in groups with milder sclerosis and serum creatinine (Cr) was elevated in groups with more severe sclerosis. Because the GTJ was assumed to disappear when an atubular glomerulus was formed, the ratio of the number of glomeruli with discernible GTJ to the total number of glomeruli was evaluated. As glomerular sclerosis progressed, discernible GTJ reduced significantly (p <0.001) and the degree of PF increased significantly (p <0.05). By serial section study in cases with pronounced PF, transitions between the stages of stenosis of the GTJ and atubular glomeruli were observed. It is speculated that the occlusion of the GTJ eventually hyalinizes the glomerulus; in such cases, glomerular obsolescence of the collapse type might be formed. On the other hand, obsolescence of the mesangial proliferative type might be formed in the hyalinization derived from direct glomerular injury. In this context, glomerular obsolescence of the collapse type was observed more frequently and was accompanied by more increased PF than obsolescence of the mesangial proliferative type (p <0.001). These results suggest that in addition to direct glomerular injury, nephron injury derived from interstitial damage of this type plays an important contributory role in the progression of IgAN. PMID- 12081199 TI - Short-term preoperative radiotherapy interferes with the determination of pathological parameters in rectal cancer. AB - Short-term preoperative radiotherapy in combination with surgery has been shown to decrease the rate of local recurrence in rectal cancer patients. The effects of this type of radiotherapy on the histopathology of rectal carcinoma has been hitherto unknown. Since various histopathological factors are associated with prognosis, the study of alterations induced by irradiation is an important issue. This paper examines the histopathology of resection specimens from 1306 patients who were treated in a randomized trial that evaluated the benefits of preoperative radiotherapy. In this trial, patients were treated with short-term radiotherapy (5 x 5 Gy) and operated on within 5 days after radiation. Histopathological parameters were determined by the Pathology Review Committee of the trial and we compared tumours of patients with and without preoperative radiotherapy. Tumours of patients who were treated with preoperative radiotherapy were smaller, more often mucinous carcinomas (13% versus 7%, p < 0.001) and more often poorly differentiated (35% versus 24%, p<0.001). After radiotherapy, there was less inflammatory reaction around the tumour (extensive in 7% versus 18%, p<0.001), which was mainly caused by a decrease in T lymphocytes and neutrophil granulocytes. The fibroblastic reaction was more pronounced in the radiotherapy group (extensive in 22% versus 10%, p <0.001). Remarkable histological alterations occurred within a week after 5 days of irradiation of rectal carcinomas. The prognostic value of these factors therefore needs to be re evaluated for irradiated patients. PMID- 12081200 TI - Mucin expression in the ileoanal reservoir reflects incomplete mucosal adaptation. AB - Restorative proctocolectomy is regarded as a standard surgical procedure for patients who require a proctocolectomy for ulcerative colitis and familial adenomatous polyposis. The ileal mucosa undergoes colonic phenotypic change with time, but the extent and relevance of these changes to the long-term safety of the ileoanal pouch are unclear. The aim of this study was to study the mucin biology of this adaptive process in order to assess its extent and possible impact on pouch safety. Ileoanal pouch biopsies from a cohort of patients and normal ileal and colonic controls were subjected to histological, biochemical, histochemical, and immunohistochemical mucin analysis. Mucin sulphation and sialic acid O-acetylation were studied as parameters of colonic phenotypic change. Fifty-one patients, 16 ileal, and 22 colonic controls were studied. Seventy per cent of biopsies retained villous mucosal architecture, with no cases of dysplasia detected. Ileoanal pouch mucosal sulphation and sialic acid O acetylation did not reach colonic levels, thus indicating limited evidence for a more colonic phenotype. The data from this study suggest that colonic phenotypic change within the ileoanal reservoir is incomplete, with no cases of dysplasia detected. The degree of phenotypic change is less than in previous studies, which may support, but not prove, our hypothesis that there may be a process of reversion to an ileal type mucosa in the ileoanal reservoir with time. PMID- 12081201 TI - Expression of mucins (MUC1, MUC2, MUC5AC, and MUC6) and type 1 Lewis antigens in cases with and without Helicobacter pylori colonization in metaplastic glands of the human stomach. AB - Helicobacter pylori (H. pylori) causes gastritis and intestinal metaplasia (TM) that may evolve to gastric carcinoma. Paradoxically, IM leads to clearing of H. pylori, except for some cases in which it persists in damaging the mucosa. The objective of this study was to compare the profile of mucins and type 1 Lewis antigens in IM cases with and without H. pylori. Gastric biopsies (n=32) were double-stained using immunohistochemistry (MUC1, MUC2, MUC5AC, MUC6, Le(a), sialyl-Le(a), and Le(b)) and histochemistry for H. pylori. H. pylori was observed in association with IM in 4 of 22 biopsies with IM (complete IM - 6; incomplete IM - 16). The four biopsies with IM and H. pylori displayed a particular pattern of incomplete IM: expression of MUC1 and MUC5AC and little/no expression of MUC2. The 18 biopsies with IM and without H. pylori had high levels of MUC2 expression, regardless of the IM type. The pattern of expression of type 1 Lewis antigens was similar in IM, regardless of the presence or absence of H. pylori. It is concluded that H. pylori is able to colonize incomplete IM whenever it contains foci expressing MUCI and MUC5AC and no MUC2, independently from Le(a), sialyl Le(a) and Le(b). The results suggest, furthermore, that MUC2 expression affects the ability of H. pylori to colonize IM areas, regardless of the levels of expression of MUC1 and MUC5AC. PMID- 12081202 TI - Radiotherapy and rectal cancer. AB - Pre-operative radiotherapy is being used to reduce local recurrence and improve survival following surgery for rectal cancer. A report by Nagtegaal et al. analyses the histopathological effects in a large, randomized study employing short course radiotherapy. In future, changes in some of these tumour parameters will have to be considered when auditing rectal cancer resections and identifying prognostic factors. PMID- 12081203 TI - Loss of heterozygosity at 18q21 region in gastric cancer involves a number of cancer-related genes and correlates with stage and histology, but lacks independent prognostic value. AB - Several studies support a role of 18q21 LOH, involving the DCC locus, in colorectal cancer progression; however, its contribution to the natural history of gastric cancer is less clear. Recently, a number of cancer-related genes have been mapped in the 18q21 region, either centromeric or telomeric to DCC. This study searched for 18q21 LOH in 161 gastric cancers representative of all tumour stages and main histological types. To this purpose, seven highly polymorphic markers were used flanking the 18q21 band and spanning the entire region. Thirty four out of 147 (23.1%) informative cases showed LOH. In 27 of 34 cases (79%), LOH involved all the informative loci. The remaining seven cases showed LOH at more telomeric sites and retained heterozygosity at more centromeric markers, mostly those proximal to the DCC gene. A strong correlation between 18q21 LOH and level of gastric wall invasion, lymph node metastases, or stage was found in cohesive (glandular+solid) and mixed tumours, but not in diffuse cancers. Cox univariate and multivariate analysis showed that invasion level, lymph node metastases, distant metastases, TNM stage, and histology were effective predictors of survival, whereas 18q21 LOH did not show predictive power. The simultaneous deletion of a variety of cancer-related genes with different and even opposite roles might explain why, apparently, 18q21 LOH does not per se contribute significantly to the natural history of gastric cancer, despite strong correlation with stage. PMID- 12081204 TI - BAG-i expression in human breast cancer: interrelationship between BAG-1 RNA, protein, HSC70 expression and clinico-pathological data. AB - BAG-1 (BCL-2 athanogene-1), a multifunctional protein which associates with steroid hormone receptors (including the oestrogen receptor) and the anti apoptotic BCL-2 protein, regulates steroid hormone-dependent transcription and apoptosis. Direct interaction with 70 kD heat-shock proteins, HSC70 and HSP70, may mediate the diverse functions of BAG-1. Immunohistochemistry was used to examine the expression of BAG-1 and HSC70 in 160 cases of invasive breast cancer. BAG-1 was expressed in 92% of cases; most tumours exhibited cytoplasmic BAG-1, while a smaller proportion also had nuclear immunostaining. There was a significant inverse correlation between histological grade and nuclear BAG-1 expression, with higher-grade tumours tending to have reduced nuclear BAG-1 expression, but there was no association with cytoplasmic BAG-1. There was also no significant correlation between nuclear or cytoplasmic BAG-1 expression and oestrogen receptor positivity. Since BAG-1 may be influenced by hormonal background, the relationship between grade and oestrogen receptor was examined separately in pre-menopausal and post-menopausal women. The statistically significant correlation between nuclear BAG-1 expression and low tumour grade was strong in pre-menopausal, but not apparent in postmenopausal women. A statistically significant correlation was observed between cytoplasmic, but not nuclear, BAG-1 expression and oestrogen receptor status in pre-menopausal, but not postmenopausal, women. There was no correlation between BAG-1 protein expression and RNA, suggesting that important post-transcriptional mechanisms control BAG-1 expression in vivo. HSC70 was also detected in the majority (97%) of cases, although expression was not correlated with BAG-1 levels, oestrogen receptor status or tumour grade. Overall survival in cases with high levels of nuclear BAG-1 expression was improved, though not significantly. These results are consistent with the hypothesis that BAG-1 plays an important but variable role in breast cancers developing in pre-menopausal and post-menopausal women. PMID- 12081205 TI - Does MSI-low exist? AB - Microsatellite instability is a well-recognised phenomenon. Ten to 15% of sporadic colorectal cancers with a high level of MSI form a well defined group with distinct clinicopathological features. The set of tumours with low level of microsatellite instability (MSI-low), though widely referred to, is not a clearly defined group. The definitions of MSI-low have varied among groups and between different studies from the same group. Some studies have found associations between the MSI-L phenotype and molecular features, notably a higher frequency of K-ras mutations, and, possibly, methylation of methylguanine methyltransferase. Two recent independent studies, however, showed respectively that 68% and 79%, non-MSI-H cancers showed some MSI and could therefore be classed nominally as MSI L. There was no evidence for a qualitatively discrete MSI-L group, but quantitative differences in the level of MSI were found. PMID- 12081206 TI - Deregulated Bag-1 protein expression in human oral squamous cell carcinomas and lymph node metastases. AB - Bag-1 is an anti-apoptotic protein that promotes metastasis in some tumour cell types. To determine whether Bag-1 expression is altered in 64 oral squamous cell carcinomas, tumour samples were compared with 17 samples of normal oral epithelium. Normal oral epithelia had pronounced nuclear staining in the basal and maturation layers and weak cytoplasmic staining that was most pronounced in the basal and suprabasal layers. Oral squamous cell carcinomas demonstrated a tendency for reduced nuclear staining intensity (p=0.036). Cytoplasmic staining intensity was not significantly different between tumour and normal tissue. However, many tumours were observed to have less of a difference between nuclear staining intensity and cytoplasmic staining intensity than normal oral epithelium. Furthermore, in lymph node metastases, cytoplasmic Bag-1 staining was stronger in 8/13 cases than in corresponding primary tumours (p=0.021). Western blotting using nine oral primary carcinoma cell lines and four normal keratinocyte cultures showed that the isoforms Bag-1s, Bag-1M, and Bag-1L were expressed in normal and malignant oral epithelial cells. Bag-1L unique sequences were shown to adopt an exclusively nuclear, and predominantly nucleolar, localization by use of transiently transfected N-terminal Bag-1L-EGFP. However, levels of Bag-1L in carcinoma cells did not differ significantly from those of normal keratinocytes. Therefore the reduced nuclear staining observed in oral squamous cell carcinomas compared with normal epithelium may reflect changes in the localization of Bag-1 isoforms, rather than decreased expression of Bag-1L. Alterations in the relative proportions of Bag-1S, Bag-1M, and Bag-1L were detected in 6/9 oral carcinoma cell lines; 5/9 oral carcinoma cell lines had a significantly greater proportion of Bag-1M than normal keratinocytes and in another cell line, Bag-1L was significantly underrepresented. Overall, the results suggest that Bag-1 deregulation plays a role in oral carcinogenesis at two different stages: during primary carcinoma development and during lymph node metastasis. PMID- 12081207 TI - Expression and regulation of collagenase-2 (MMP-8) in head and neck squamous cell carcinomas. AB - MMP-8 (collagenase-2) is the most effective collagenase to initiate type I collagen degradation. Since initiation of lysis of the surrounding collagen matrix is an essential prerequisite for carcinoma cells to spread, this study investigated the expression of MMP-8 in squamous cell carcinoma (SCC) of the head and neck in vivo and in vitro. Most of the recently established head and neck carcinoma cell lines (22/25), corresponding tumour (5/7) and dermal (2/2) fibroblasts, commercial tongue carcinoma (HSC-3 and SCC-25), and transformed keratinocyte cell lines of the tongue (IHGK) and skin (HaCaT) expressed MMP-8 mRNA analysed by the PCR method. Western blotting revealed a latent 50 kD band in concentrated culture media of carcinoma cells and corresponding tumour and dermal fibroblasts. The expression of immunoreactive MMP-8 protein was reduced 30% by transforming growth factor beta-1 (TGF-beta1) at 1 ng/ml concentration and 60% at 10 ng/ml concentration, but up-regulated 2- and 2.5-fold after 10 nM and 100 nM phorbol 12-myristate 13 acetate (PMA), respectively. Immunohistological staining localized MMP-8 protein in a few malignant invading tumour cell islands, certain fibroblasts, polymorphonuclear neutrophils (PMNs), and plasma cells. In situ hybridization revealed a faint sporadic signal in carcinoma cells of all eight tissue sections analysed. It is concluded that tissue from head and neck carcinomas can express MMP-8 both in vivo and in vitro. Since the amount of MMP-8 in carcinoma and stromal cells is rather low, MMP-8 may have a potential role, with other collagenases, in the proteolysis of connective tissue associated with the spreading of invasive carcinoma. PMID- 12081208 TI - Frequent phosphorylation at serine 392 in overexpressed p53 protein due to missense mutation in carcinoma of the urinary tract. AB - Post-transcriptional modification of p53 by phosphorylation has been proposed to be an important mechanism of p53 stabilization and functional regulation. Phosphorylation of p53 Ser392, in particular, activates specific DNA binding functions by stabilizing p53 tetramer formation. This study evaluated the relationship between p53 Ser392 phosphorylation and various types of p53 missense mutation detected in urothelial transitional cell carcinomas (TCCs), with stratification of the mutations according to the functional domains elucidated by the crystal structure of the p53 protein. Of 41 TCCs with missense mutations, 26 (63.4%) exhibited immunopositivity with Ser392 phospho-specific p53 antibody. In comparison to structural mutations, the missense mutations at exon 7 (p=0.0307) or located in regions that affect direct DNA binding ability (p = 0.0273) were significantly associated with Ser392 immunopositivity. No statistically significant relationship was found between Ser392 immunoreactivity and other different types of p53 mutation. The prevalence of cases exhibiting Ser392 positive immunostaining was higher for high-grade (p <0.0001) and advanced-stage TCCs (p =0.0119) than for TCCs with wild-type p53. No significant relationship was found between Ser392 immunoreactivity and apoptotic index in urothelial TCCs. These in vivo findings indicate that Ser392 phosphorylation frequently occurs in mutant form p53 in TCCs. Because mutant form p53 can act dominant-negatively by heterooligomerization with wild-type p53, these findings also suggest that Ser392 phosphorylation might activate tetramer formation to promote the dominant negative effects of mutant form p53, and thereby contribute to proliferation of aggressive TCCs. PMID- 12081210 TI - Frequent genomic imbalances in chromosomes 17, 19, and 22q in peripheral nerve sheath tumours detected by comparative genomic hybridization analysis. AB - Comparative genomic hybridization (CGH) was used to detect changes in relative chromosome copy number in 50 cases of peripheral nerve sheath tumour (PNSTs), including nine malignant peripheral nerve sheath tumours (MPNSTs), 27 neurofibromas (with three plexiform neurofibromas) and 14 schwannomas. Chromosome imbalances were frequently detected in benign as well as malignant PNSTs. In both NF1-associated and sporadic MPNSTs, the number of gains was higher than the number of losses, suggesting proto-oncogene activation during MPNST progression. NF1-asociated MPNSTs exhibited gains of chromosomes 17q and X (2/4 cases each), whereas sporadic MPNSTs showed gains of chromosome 4q (3/5 cases). On the other hand, in benign neurofibromas and schwannomas, the number of losses was higher than the number of gains, suggesting a predominant role of tumour suppressor genes in tumourigenesis. Both sporadic and NF1-associated neurofibromas exhibited losses at chromosome 22q in more than 50% of cases. These chromosomal regions may contain common chromosomal abnormalities characteristic of both types of neurofibromas. In NF1-associated neurofibromas, most frequent losses were found in chromosomes 17 [17p11.2-p13 in nine cases (60%); 17q24-25 in 6 cases (40%)] and 19 [19p13.2 in eight cases (53%); 19q13.2-qter in eight cases (53%)], whereas in sporadic neurofibromas and schwannomas losses of chromosomes 17 and 19 were detected in less than 50% of cases. Since this 17p11.2-p13 region is known to contain the tumour suppressor gene TP53, patients with NF1 may be at high risk of malignant neoplasms including MPNSTs. Gains were more frequently detected in plexiform neurofibromas (2/3 cases) than other benign tumours, suggesting proto oncogene activation in tumourigenesis of plexiform neurofibroma. The significance of the losses of chromosome 19 in these cases is not clear at present, but in NF1 associated neurofibromas, the presence of some as yet unknown tumour suppressor genes on chromosome 19 cannot be ruled out. PMID- 12081209 TI - Expression of CXC chemokine IP-10 in testicular germ cell tumours. AB - Tumour-infiltrating T lymphocytes (TILs) possess discrepant properties ranging from anti- to protumour activities. Understanding precisely which mechanisms navigating T lymphocytes into the tumour site will help to further the anti tumour or to disrupt the pro-tumour activities of TILs. The present study asked what enables TILs to migrate into testicular germ cell tumours (TGCTs). TILs were characterized and the expression of a large panel of T-lymphocyte-attracting chemokines was investigated in 21 TGCT cases. Flow cytometry revealed that approximately 80% of TGCT-infiltrating T lymphocytes express CXCR3, a receptor for the chemokine interferon-inducible protein-10 (IP-10). RT-PCR and immunohistochemistry indicated that IP-10 was the only chemokine investigated which was constantly expressed in TGCT. As IP-10 was found to be expressed by endothelial cells of TGCT-associated blood vessels, the question arose whether the IP-10-regulating cytokine interferon-gamma (IFNgamma) is produced by tumour cells and if so, whether tumour-derived IFNgamma can induce IP-10 in endothelial cells. Applying in situ hybridization, IFNgamma transcripts were found in neoplastic germ cells. Analyses of two TGCT cell lines indicated that the tumour cells not only express IFNgamma mRNA, but also produce and secrete IFNgamma protein; tumour-derived IFNgamma provokes IP-10 expression and secretion by endothelial cells in vitro, as assessed by PCR and ELISA. Together, the data suggest that neoplastic germ cells secret IFNgamma and thereby stimulate tumour associated endothelial cells to express IP-10, which contributes to the recruitment of CXCR3+ T lymphocytes to the site of TGCTs. PMID- 12081211 TI - Osteoporosis, the deafening silent epidemic. PMID- 12081212 TI - Osteoporosis 2002: headline news. PMID- 12081213 TI - Bisphosphonates, statins, osteoporosis, and atherosclerosis. PMID- 12081214 TI - Vertebroplasty and kyphoplasty. PMID- 12081215 TI - Osteoporotic fractures. PMID- 12081216 TI - Selected annotated bibliography. PMID- 12081217 TI - Selected ongoing clinical trials. PMID- 12081218 TI - Recommendations for prevention and management of fire ant infestation of health care facilities. AB - The range of imported fire ants now includes most of the southeastern United States, parts of the Southwest, portions of the East and West Coasts, and Puerto Rico. Increasingly, fire ant attacks on patients in health care facilities have been reported. In this paper, we provide recommendations for fire ant control in and around health care facilities that should help prevent building infestation and further attacks. In addition, we provide algorithms detailing fire ant prevention strategies (indoors and outdoors) and patient management after fire ant stings. Physicians in areas endemic for fire ants should be aware of the possibility that patients may be harmed by these insects, and also should be generally familiar with measures used to control fire ants. PMID- 12081219 TI - Evaluation of a portable clinical analyzer in the pediatric emergency department: analysis of cost and turnaround time. AB - BACKGROUND: The objective of this study was to evaluate a portable clinical analyzer (PCA) in the pediatric emergency department (ED), examining (1) turnaround time, (2) cost, and (3) sample amounts for PCA versus standard laboratory. METHODS: Twenty children were studied. Laboratory measurements were taken from the study group using PCA and central laboratory as the control. RESULTS: The PCA turnaround times were 54.2 minutes faster than the central laboratory. Cost was $1.65 less for the PCA. The PCA sample size was smaller (0.29 mL vs 2.62 mL). CONCLUSIONS: Laboratory values obtained by PCA were available to treating physicians significantly faster than those from the central laboratory. The PCA costs less and uses a smaller blood sample than the central laboratory. Use of PCA technology has the potential to decrease laboratory turnaround time and cost. Further investigation on the influence of patient care is needed. PMID- 12081220 TI - Community-wide medical entomology training for medical practitioners. PMID- 12081221 TI - Sports medicine approach to low back pain. AB - Low back pain is a common, recurrent affliction in the general population. Research has indicated that, contrary to traditional beliefs, bed rest is not an effective therapy. Existing evidence supports the use of early activity and exercise in treatment of acute and chronic low back pain. Since the sports medicine model of rehabilitation is based on early, progressive activity, its principles can be usefully applied to the management of low back pain in the general population. PMID- 12081222 TI - Spontaneous remission of B-cell chronic lymphocytic leukemia associated with T lymphocytic hyperplasia in bone marrow. AB - Spontaneous remissions in B-cell chronic lymphocytic leukemia (B-CLL) are rare and none of them has been studied with immunophenotyping (by flow cytometry and immunohistochemistry) and genotyping. In this patient, studied after spontaneous remission had occurred, there was a residual T-lymphocytic hyperplasia in the bone marrow with a normal CD4:CD8 ratio. Absolute CD4 and CD8 counts and CD4:CD8 ratio in the peripheral blood were normal. Flow cytometry revealed no B-CLL cells in the peripheral blood and less than 2% B-CLL cells in the bone marrow. PMID- 12081223 TI - Severe hepatitis associated with oxacillin therapy. AB - A 6-year-old girl had fever, abdominal pain, and severe anicteric hepatitis during intravenous oxacillin therapy for staphylococcal osteomyelitis. She had greatly elevated liver enzymes, prolonged prothrombin time, leukopenia, and eosinophilia. Clinical symptoms resolved and laboratory data returned to normal after withdrawing oxacillin and substituting cefazolin. This hepatotoxicity appears to be specific to oxacillin and not to other beta-lactams. Monitoring liver function tests during oxacillin therapy, especially in patients receiving prolonged treatment, may be warranted. PMID- 12081224 TI - Person-to-person transmission of Pseudomonas pneumonia in the community: documentation by pulsed-field electrophoresis. AB - Pseudomonas aeruginosa is a rare cause of community-acquired infection. The source of this organism has usually been inapparent or environmental (ie, contaminated humidifiers). We documented transmission of P aeruginosa leading to cavitary pneumonia and lung abscess from daughter to mother and confirmed the clonal identity of our two patients' isolates using pulsed-field electrophoresis. PMID- 12081225 TI - Highlights from the annual scientific assembly: mechanisms to stop the epidemic of obesity: surgical therapy for obesity. PMID- 12081226 TI - Legionella pneumonia: an unusual cause of rhabdomyolysis and acute renal failure. PMID- 12081227 TI - Unfiltered coffee raises cholesterol. PMID- 12081228 TI - Transgenic approach toward leptin biology: the clinical implications of leptin for the treatment of obesity-associated diabetes and obesity-related hypertension. PMID- 12081229 TI - Regulation of the insulin gene transcription by glucose. PMID- 12081230 TI - Sex-related differences in gene expression in neonatal rat hypothalamus assessed by cDNA microarray analysis. AB - Sexual differentiation of the rodent brain is recognized to involve transcriptional activation of multiple genes induced by gonadal steroids at developmental stages. To identify the genes differing in expression level between sexes, we analyzed gene expression in male and female rat hypothalami at postnatal day 5 by means of a cDNA microarray consisting of 2352 genes. By comparing the expression pattern between sexes, we identified 12 male-enriched genes and 20 female-enriched genes. Among them, the expression pattern of 1 male enriched gene, jagged homolog 1, and those of 2 female-enriched genes, p27Kip1 and p130, were confirmed to be consistent with microarray data by RT-PCR. Investigation of these genes should help to elucidate the molecular and cellular mechanisms underlying sexual differentiation of the rodent central nervous system. PMID- 12081231 TI - Effect of octreotide acetate on the plasma concentration and urinary excretion of uridine and purine bases. AB - To determine the effect of octreotide acetate on urinary excretion of uric acid and plasma concentration of uridine, we subcutaneously administered octreotide acetate (1 microg/kg of body weight) to 5 healthy subjects. Ninety minutes after administration, octreotide acetate increased the plasma concentration of uridine by 15% and decreased the plasma concentration of glucagon by 24% and that of insulin to below the detection limits. In addition, octreotide acetate decreased the urinary excretion of uric acid, sodium, and chloride by 60%, 40%, and 38%, respectively, at 1 hour after administration. However, octreotide acetate did not affect the concentrations of hypoxanthine, xanthine, uric acid, cyclic AMP in plasma, lactic acid and pyruvic acid in blood, urinary excretion of hypoxanthine and xanthine, or creatinine clearance. From these results, we speculated that octreotide acetate decreases the urinary excretion of uric acid by decreasing the concentration of glucagon and/or urinary excretion of sodium, and increases the plasma concentration of uridine via decreased concentrations of glucagon and insulin. PMID- 12081232 TI - Characterization of GH pulsatility in male Shiba goats: effects of postpubertal castration and KP102. AB - The present study was conducted in order to characterize the secretory pattern of GH in the Shiba goat, a native Japanese miniature goat, and to examine the effects of castration and KP102, a GH secretagogue, on this pattern. Blood samples were taken from an indwelling jugular catheter every 15 min for 24 h, and plasma GH was measured by radioimmunoassay. In intact males, GH was secreted in a pulsatile manner with very regular 5-h periodicity, which consisted of a distinctive GH pulse and a trough of virtually no GH secretion. Postpubertal castration increased the height and decreased the width of GH pulses, though it did not affect the interpulse interval and area under the curve. Modification of the shape of each GH pulse by testicular androgen might play a role in the expression of GH action in the male. KP102 (10 microg/kg, i.v.) immediately induced a robust GH pulse, which was followed by a spontaneous GH pulse of normal characteristics at regular intervals, suggesting that the clock generating GH pulses was reset by KP102. From these observations, we concluded that the Shiba goat is a very suitable experimental model for elucidating the mechanisms underlying GH pulse generation, and in particular, the involvement of androgen and GH secretagogues. PMID- 12081233 TI - Adrenocorticotropin-producing pituitary carcinoma with metastasis to the liver in a patient with Cushing's disease. AB - A 61-year-old woman who had a nonfunctioning pituitary tumor removed was later found to have a pituitary carcinoma with metastasis to the liver that produced adrenocorticotropic hormone (ACTH), causing Cushing's disease. Despite the second removal of the pituitary tumor, Cushing's disease persisted. Endocrinologic examination demonstrated marked elevation of plasma ACTH, beta-endorphin, and serum cortisol concentrations, which were not suppressed by low- or high-dose dexamethasone administration. A computed tomographic scan of the abdomen revealed a solitary hypodense area in the liver consistent with a metastatic lesion. Immediately after resection of the liver tumor, plasma ACTH concentrations fell to normal range. Such a decline in ACTH secretion was not observed even after the pituitary tumor was removed. Histologic characteristics of both the pituitary and liver tumors were similar. The immunohistochemical study using anti-ACTH antibody showed similar ACTH-positive cells in the pituitary and liver tumors. Based on these findings, the patient was diagnosed as having liver metastasis of ACTH secreting pituitary carcinoma. This is the first case report in which a metastatic liver tumor of a pituitary carcinoma caused ACTH. PMID- 12081234 TI - A modified hMG-GnRH method for the induction of ovulation in infertile women with severe hypogonadotropic amenorrhea. AB - The objective of this study was to compare, in infertile women suffering from severe hypogonadotropic amenorrhea, the therapeutic utility and the incidence of complications arising from fertility treatment by the conventional human menopausal gonadotropin/human chorionic gonadotropin (hMG-hCG) method, the hMG step-down method, the sequential hMG/gonadotropin-releasing hormone (GnRH) method and a new, modified hMG-GnRH method that has been developed by us. In the step down method, the daily dose of hMG was decreased from 150 IU to 75 IU when the follicle diameter reached 11-13 mm. In the sequential hMG-GnRH, hMG injection was switched to pulsatile GnRH administration (20 microg/120 min SC), when the follicle diameter reached 11-13 mm. In our new modified hMG-GnRH, pulsatile GnRH was injected together with hMG. Daily hMG was stopped and the GnRH dosage was changed from 10 microg to 20 microg when the follicle diameter reached 11-13 mm. Initially, the three established methods were applied randomly to treat 34 cycles in 20 women; and subsequently, five patients who failed to conceive following treatment by sequential hMG-GnRH were then treated by the modified hMG-GnRH method. More than eight growing follicles and multiple pregnancies were observed during treatment by the conventional method. The incidence of ovarian hyperstimulation syndrome (OHSS) was 25.7% with the conventional method, 20.0% with the step-down method and 0% with the sequential hMG-GnRH method; however, the rate of ovulation was only 50% with the sequential hMG-GnRH method. By contrast, with the modified hMG-GnRH method, less than three growing follicles occurred in 81.8% of patients, there was a 100% rate of ovulation, and neither OHSS nor multiple pregnancies were observed. Moreover, the modified hMG-GnRH method induced pregnancy in 3 out of 5 patients. These data indicate that this new method is favorable for the treatment of severe hypogonadotropic amenorrhea. PMID- 12081235 TI - Bright pituitary stalk on MR T1-weighted image: damming up phenomenon of the neurosecretory granules. AB - Characteristic findings of the pituitary stalk on magnetic resonance (MR) imaging, which suggest a damming-up phenomenon of neurosecretory granules, were reported. Neurosecretory granules containing vasopressin influence the signal intensity on MR T1-weighted image (T1WI). The normal posterior lobe of the pituitary gland appears as a bright signal on T1WI. The bright signal of the posterior lobe represents the normal content of neurosecretory granules and disappears in patients with central diabetes insipidus. The normal pituitary stalk appears as a low-intermediate intensity signal on sagittal and coronal T1WIs with 3 mm-slice thickness. The pituitary stalk appeared as a bright signal in 20 patients; 13 with pituitary adenoma, 4 with an intrasellar cystic lesion, one with cavernous sinus mass, and 2 with no abnormal MR findings. The pituitary stalk was not severed in any of the cases. The normal bright signal of the posterior lobe disappeared in 17 patients. No patients suffered from symptoms of central diabetes insipidus when the bright pituitary stalk appeared. It is suggested that the origin of the bright signal in the pituitary stalk is the damming up and accumulation of neurosecretory granules in the nerve fibers of the hypothalamohypophyseal tract obstructed by adenoma, postoperative scarring, cystic mass and so on. Probably, the damming-up phenomenon on MR imaging represents the functional integrity of the hypothalamo-neurohypophyseal system, and should be distinguished from an ectopic posterior lobe formation which is caused by stalk transection. PMID- 12081236 TI - Multiple messenger ribonucleic acid transcripts and revised gene organization of the human TSH receptor. AB - Northern blot analysis of human TSH receptor (hTSHR) messenger ribonucleic acid (mRNA) expression has previously demonstrated multiple species of transcripts in the thyroid gland, suggesting the presence of multiple transcription initiation sites, alternatively spliced forms or alternate polyadenylation (poly(A)) sites. The first two have already been reported elsewhere. To clarify alternate poly(A) sites in the hTSHR gene, the present study was designed to characterize three full-length hTSHR cDNAs with distinct poly(A) signals that we have previously cloned. The comparison of the nucleotide sequencing data on the 3'UTR of these three clones to the Draft Human Genome in NCBI database revealed that the 3' segment of exon 10 of hTSHR gene contains three tandem repeats of the poly(A) sites, from which are expressed three full-length TSHR mRNAs with distinct 3'UTR length. The longest one appears to be a predominant transcript. From these data, together with (i) the previously reported organization of hTSHR genome and (ii) use of the Draft Human Genome to localize the unidentified sequence in the alternatively spliced form of truncated hTSHR, we propose the complete structure of hTSHR gene. Rather than 10 exons, our analysis suggests that hTSHR gene seems to contain 13 exons and 12 introns. At least three full-length TSHR mRNAs with distinct poly(A) sites and five alternatively spliced forms of TSHR mRNAs are expressed from the single hTSHR gene. PMID- 12081237 TI - A case of von Hippel-Lindau disease with bilateral pheochromocytoma, renal cell carcinoma, pelvic tumor, spinal hemangioblastoma and primary hyperparathyroidism. AB - A rare case of von Hippel-Lindau (VHL) disease with bilateral pheochromocytomas, right renal cell carcinoma, right pelvic carcinoma, spinal hemangioblastoma and primary hyperparathyroidism is described. A 78-year-old woman had a history of hypertension from her forties. She suffered from headache and body weight loss. Abdominal CT revealed bilateral adrenal tumors and right external renal tumors enhanced in early stage. MIBG scintigraphy exhibited a high accumulation of tracer in both adrenal glands. On the basis of the radiographic findings and endocrinological results, the patient was diagnosed as having bilateral pheochromocytomas and right renal cell carcinoma. A bilateral adrenectomy was performed, followed by surgery for resection of the renal cell carcinoma. The other resected right kidney showed a clear cell subtype that was determined to be renal cell carcinoma, and proved that the pelvic tumor was transient cell carcinoma. Spinal MRI showed spinal hemangioblastoma. von Hippel-Lindau (VHL) gene mutation for the patient was found. We diagnosed the patient as VHL because of the existence of spinal hemangioma and a VHL disease gene. Parathyroid echo revealed a hypoechoic space on the back of the left lobe, and serum calcium and intact PTH to be elevated. The patient was diagnosed as primary hyperparathyroidism. We report the first case of a patient with VHL disease complicated with bilateral pheochromocytomas, right renal cell carcinoma, right renal pelvic carcinoma and primary hyperparathyroidism. The life expectancy of affected individuals has been less than 50 years. Since the prognosis may be improved by an early diagnosis, affected individuals with VHL complexes should undergo cranial, spinal MRI and abdomen CT. The families may benefit from presymptomatic detection of affected gene carriers and the exclusion of at-risk family members by negative test results. PMID- 12081238 TI - Accelerated production of nucleosome in cultured human mononuclear cells in untreated Graves' disease. AB - The apoptosis of lymphocytes, which occurs in autoimmune diseases, is usually induced by the Fas/Fas ligand system. As the assay of nucleosomes produced by apoptotic cells can be used to quantitate apoptosis, we evaluated nucleosome and soluble Fas ligand (sFasL) levels of cultured mononuclear cells to clarify the apoptosis of mononuclear cells in patients with autoimmune thyroid diseases by enzyme-linked immunosorbent assay. Nucleosome levels of cultured mononuclear cells in patients with untreated Graves' disease were significantly higher (3.27+/-2.90 U/ml) than those of control subjects (1.39+/-0.24 U/ml) and euthyroid patients with treated Graves' disease (1.53+/-0.33 U/ml). Nucleosome levels of cultured mononuclear cells were positively correlated with sFasL levels (r=0.544, p<0.01). It is therefore likely that increased sFasL levels elicit apoptosis of these cells in untreated Graves' disease. PMID- 12081240 TI - Familial occurrence of subacute thyroiditis. AB - Although subacute thyroiditis (de Quervain's thyroiditis) is presumed to be caused by a viral infection, only 2 familial occurrences of subacute thyroiditis have been reported in the literature. Typical and severe subacute thyroiditis was diagnosed in an older sister who was hospitalized for 8 days. During this period her younger sister nursed her. Three weeks after, the same clinical picture occurred in the younger sister. We postulate that subacute thyroiditis might occur by transmission of possible viral infection in genetically predisposed individuals. PMID- 12081239 TI - Temporal expression of type I interferon receptor in the peri-implantation ovine extra-embryonic membranes: demonstration that human IFNalpha can bind to this receptor. AB - Interferon-tau (IFNtau), produced by the trophectoderm of ruminant ungulates, binds to the type I IFN receptor (IFNAR) located at the uterine endometrium in a paracrine manner. Since IFNtau attenuates the secretory pattern of an endometrial luteolysin, prostaglandin F2alpha, IFNtau has been considered as a conceptus factor implicated in the process of maternal recognition of pregnancy. Here we report the presence of IFNAR subunit (IFNAR1) in ovine conceptuses during the period of peri-implantation development and demonstrate that 125I-human (h) IFNalpha binds to membrane preparations from ovine corpus luteum and conceptus. Using an antibody against hIFNAR1, immunohistochemical analysis revealed that IFNAR1 protein was present in day 14 and 16 conceptuses (day 0 = day of estrus) and luminal and glandular epithelia of the endometrium. Conceptus membrane proteins analyzed by western blot with the same antibody displayed immunoreactive bands at 95, 60 and 55 kDa while endometrial membrane proteins showed bands at 200, 95 and 55 kDa. Northern blot analysis revealed that IFNAR1 mRNA was present in days 15-19 conceptuses and day 18-19 allantoic membranes. Receptor binding studies indicated that 125I-hIFNalpha binding to day 16, but not earlier, conceptus membrane proteins could be displaced with hIFNalpha or ovine IFNtau. Based on Scatchard analysis, day 16 conceptus membranes contained 28 fmol IFNAR/mg protein with a dissociation constant of 300 pM. Cross-linking experiments demonstrated that 125I-hIFNalpha-receptor complex migrated at 120 kDa, indicating that the receptor component(s) was approximately 100 kDa. These data provide evidence that although the binding does not occur until day 16, ovine conceptuses possess IFNAR1 near or at the time of implantation, suggesting that IFNtau, a factor produced by the trophectoderm of ruminant ungulates, could act on the conceptus in an autocrine manner. In addition to functioning as an antiluteolytic factor, therefore, IFNtau may have a direct effect on conceptus development. PMID- 12081241 TI - Plasma lipids and osteoporosis in postmenopausal women. AB - Many clinical studies have shown that osteoporosis is associated with atherosclerosis and cardiovascular death. Although both high plasma levels of low density lipoprotein cholesterol (LDL-C) and low plasma levels of high density lipoprotein cholesterol (HDL-C) are known to be risk factors for atherosclerosis, it is unclear whether such lipid derangements are also associated with the pathogenesis of osteoporosis. In this study, we evaluated the relationships between plasma levels of total C, LDL-C, HDL-C, or triglyceride (TG) versus bone mineral density (BMD) at the lumbar spine, femoral neck, radius, or total body as well as the presence of vertebral fractures in 214 Japanese postmenopausal women (age range, 47-86 years, mean 62.7). Multiple regression analysis was performed between BMD at each skeletal site versus each lipid level adjusted for age, years after menopause, body mass index (BMI), and %fat. Plasma LDL-C levels were significantly and inversely correlated with the absolute values of both one-third radial (1/3R) and distal radial (UDR) BMD (p<0.01), and tended to be inversely correlated with the absolute values of L-BMD (p=0.051). In contrast, plasma HDL-C levels were significantly and positively correlated with the absolute values of L, 1/3R and UDR BMD (p<0.05). On the other hand, plasma TG levels were significantly lower in women with vertebral fractures than in those without fractures (97.0+/-36.5 vs. 126.4+/-65.8 mg/dl, mean+/-SD, p<0.05). When multivariate logistic regression analysis was performed with the presence of vertebral fractures as a dependent variable and each lipid level adjusted for age, years after menopause, BMI, and %fat as independent variables, TG alone was selected as an index affecting the presence of vertebral fractures (odds ratio: 0.51, 95% confidential interval: 0.29-0.89 per SD increase, p<0.05). Our study showed that plasma LDL-C and HDL-C levels were inversely and positively correlated with both R- and L-BMD values, respectively, while low plasma TG levels were associated with the presence of vertebral fractures in postmenopausal women. Thus, plasma lipids might be related to bone mass and bone fragility, and might be the common factor underlying both osteoporosis and atherosclerosis. PMID- 12081242 TI - Alterations in nitrate/nitrite and nitric oxide synthase in preovulatory follicles in gonadotropin-primed immature rat. AB - Nitric oxide synthase (NOS) and nitric oxide (NO) play important roles in ovulation. The purpose of this study was to investigate the changes of intrafollicular nitrate/nitrite concentration and NOS mRNA expression in preovulatory follicles during equine CG (eCG) and human CG (hCG) induced ovulation in immature rats. Immature Sprague-Dawley rats received 15 IU eCG and then 15 IU hCG 48 h later. Rats were killed immediately before, 5 h after or 10 h after hCG injection, and their preovulatory follicles were dissected. Follicular fluid, granulosa cell, and theca cell layers were collected from preovulatory follicles and assayed for NO or NOS mRNA or for in vitro incubation study. Nitrate/nitrite concentration in the follicular fluid decreased significantly 5 and 10 h after hCG injection. Inducible NOS (iNOS) mRNA expression, which was greater in granulosa cell than in the theca cell layer, decreased significantly 5 and 10 h after hCG injection. However, endothelial NOS (eNOS) mRNA expression was detected mainly in the theca cell layer and further increased 5 and 10 h after hCG injection but remained low in granulosa cells. In vitro treatment of granulosa cells with 10(-4) or 5x10(-4) M S-nitroso-L-acetyl penicillamine (NO donor) decreased progesterone production and increased DNA fragmentation. We concluded that the decrease in nitrate/nitrite concentration in preovulatory follicles after hCG injection was due mainly to decreased iNOS expression in granulosa cells. These changes in nitrate/nitrite concentration may prevent apoptosis in preovulatory follicles. PMID- 12081243 TI - A case of ACTH-independent bilateral macronodular adrenal hyperplasia successfully treated by subtotal resection of the adrenal glands: four-year follow-up. AB - We report ACTH-independent bilateral macronodular adrenal hyperplasia (AIMAH) seen in a 48-year-old male with a history of rectal and skin cancer. Bilateral multiple adrenal nodular lesions on abdominal CT scans, elevated early morning plasma cortisol levels with undetectable plasma ACTH, increased 24-hour urinary free cortisol excretion, and loss of the normal circadian rhythm in cortisol secretion established the diagnosis of AIMAH. Subtotal resection of the adrenal glands preserving lower one third of the left side was performed, considering the unfavorable effects of total adrenalectomy on future treatment of his malignancies. He has been doing well for four years with normal plasma ACTH and cortisol levels. This case suggests that subtotal resection of the adrenal glands may be applicable to selected patients. PMID- 12081244 TI - Late onset of adrenocortical failure in GH deficiency with invisible pituitary stalk: a case report of a 48-year-old Japanese man and review of the literature. AB - ACTH deficiency gradually develops in patients with growth hormone deficiency (GHD) who have abnormalities of the pituitary stalk on magnetic resonance imaging (MRI) following perinatal complications. We report here a rare case of GHD manifesting ACTH deficiency in middle age. A 48-year-old male patient was admitted to our hospital due to fever and hyponatremia. He was diagnosed as GHD and hypothyroidism at the age of 9, and had received lysine treatment until age 20, which was then replaced by thyroid hormone. He was not mentally retarded, but was the shortest in his class throughout his schooldays, reaching a final height of 148 cm. Hormonal examination revealed the presence of hypoadrenalism as indicated by poor responses of plasma cortisol to intravenous administration of corticotropin-releasing hormone (CRH) and insulin-induced hypoglycemia. Plasma ACTH responded well to CRH, but not to insulin-induced hypoglycemia, indicating that his hypoadrenalism was of hypothalamic origin. MRI showed an invisible pituitary stalk and relatively small pituitary gland. Since he had a perinatal abnormality, the damage around the pituitary and GHD could have originated from birth. In the literature, around 60% of GHD patients with pituitary stalk abnormalities develop hypoadrenalism due to ACTH deficiency, and more than 90% of such cases are diagnosed by age 30. We suggest that the appearance of hypoadrenalism should be carefully monitored in GHD patients with pituitary stalk abnormalities even after they reach middle age. PMID- 12081245 TI - A case of TSH receptor antibody-positive hyperthyroidism with functioning metastases of thyroid carcinoma. AB - The presence of TSH receptor antibody (TRAb) is rarely responsible for hyperthyroidism due to metastatic lesions of thyroid carcinoma. A 70-year-old woman was incidentally found to be thyrotoxic around the time that external irradiation was performed for multiple bone metastases 9 years after subtotal thyroidectomy for follicular carcinoma. Hyperthyroidism persisted after oral administration of thiamazole. Relevant laboratory data were as follows: FT4 9.6 ng/L, FT3 7.3 ng/L, TSH <0.19 mU/L, TBII 70, TSAb 735, and Tg 32,000 microg/L. 131I-total body scan showed 131I accumulation in the occipital bone, cervical vertebra, thoracic vertebra, ilium, and residual thyroid gland. Since the ilium uptake (11.6) was markedly higher compared to the residual thyroid gland uptake (0.14), four subsequent 131I therapies were performed. The patient became hypothyroid, and TBII became negative. TSAb became negative after the first 131I therapy but has increased again to 204 at present. Tg was 1,962 microg/L despite high TSH levels. 131I accumulation in the residual thyroid, cervical vertebra, and thoracic vertebra disappeared. Also 131I accumulation in the ilium has gradually decreased, but the image in the occipital bone has become markedly distinctive. This is a rare case characterized by TRAb-positive hyperthyroidism, by T3-predominant thyrotoxicosis, and by stronger accumulation of 131I in the metastatic tumor than in the residual thyroid gland. Thus, the response to TRAb and 131I-therapy is different among metastatic thyroid tissues. PMID- 12081246 TI - Cancer-specific mRNAs in thyroid carcinomas: detection, use, and their implication in thyroid carcinogenesis. AB - Molecular-based diagnosis ofthyroid carcinomas can be more easily establishedby utilizing specific mRNAs that are expressed in a restricted manner in cancer tissues. Accordingly, several cancer-specific mRNAs in thyroid carcinomas have been identified by means of sequence specific-differential display (SS-DD), serial analysis of gene expression (SAGE) and other new techniques. By using these cancer-specific mRNAs, some new methods of preoperative diagnosis of thyroid carcinomas have been developed. In one such method, Aspiration Biopsy Reverse Transcription-Polymerase Chain Reaction (ABRP), RNA is extracted from leftover cells within the needle used for fine needle aspiration biopsies (FNABs), thereby allowing cytological and molecular-based diagnoses to be performed simultaneously. ABRP provides both RNA information and a cytological diagnosis without further invasion to the patient. By ABRP detection of cancer specific mRNAs, papillary, anaplastic and medullary carcinomas and a part of malignant lymphomas can be accurately diagnosed preoperatively. It remains to be clarified why cancer-specific mRNAs, especially those that are overexpressed in fetal tissues, can clearly distinguish benign tissues from carcinomas, while genomic alternations, such as mutations in the RAS or P53 gene cannot. Further, the widely accepted hypothesis of multi-step carcinogenesis cannot explain some of the clinical and experimental findings of thyroid carcinomas. Considering these facts, we propose a novel hypothesis of thyroid carcinogenesis, the "germ cell carcinogenesis" hypothesis, in which cancer cells derive from the remnants of fetal thyroid germ cells (thyroblasts) instead of normal thyroid follicular cells. PMID- 12081247 TI - The achievements and challenges of the African Programme for Onchocerciasis Control (APOC). AB - The main strategy of APOC, of community-directed treatment with ivermectin (CDTI), has enabled the programme to reach, empower and bring relief to remote and under-served, onchocerciasis-endemic communities. With CDTI, geographical and therapeutic coverages have increased substantially, in most areas, to the levels required to eliminate onchocerciasis as a public-health problem. Over 20 million people received treatment in 2000. APOC has also made effective use of the combination of the rapid epidemiological mapping of onchocerciasis (REMO) and geographical information systems (GIS), to provide information on the geographical distribution and prevalence of the disease. This has led to improvements in the identification of CDTI-priority areas, and in the estimates of the numbers of people to be treated. A unique public-private-sector partnership has been at the heart of APOC's relative success. Through efficient capacity-building, the programme's operations have positively influenced and strengthened the health services of participating countries. These laudable achievements notwithstanding, APOC faces many challenges during the second phase of its operations, when the full impact of the programme is expected to be felt. Notable among these challenges are the sustainability of CDTI, the strategy's effective integration into the healthcare system, and the full exploitation of its potential as an entry point for other health programmes. The channels created for CDTI, could, for example, help efforts to eliminate lymphatic filariasis (which will feature on the agenda of many participating countries during APOC's Phase 2). However, these other programmes need to be executed without compromising the onchocerciasis-control programme itself. Success in meeting these challenges will depend on the continued, wholehearted commitment of all the partners involved, particularly that of the governments of the participating countries. PMID- 12081248 TI - Rapid epidemiological mapping of onchocerciasis (REMO): its application by the African Programme for Onchocerciasis Control (APOC). AB - One of the fundamental challenges that the African Programme for Onchocerciasis Control (APOC) has had to face is how to identify the endemic communities where its mass ivermectin-treatment operations are to be carried out in conformity with its stated objective of targetting the most highly endemic, affected and at-risk populations. This it has done by adopting a technique, known as the rapid epidemiological mapping of onchocerciasis (REMO), that provides data on the distribution and prevalence of onchocerciasis. Integration of the REMO data into a geographical information system (GIS) enables delineation of zones of various levels of endemicity, and this is an important step in the planning process for onchocerciasis control. Zones are included in (or excluded from) the APOC-funded programme of community-directed treatment with ivermectin (CDTI), depending on whether or not their levels of onchocercal endemicity reach the threshold set by APOC. This review describes the application of the REMO/GIS technique by APOC in its operations, and identifies the remaining related challenges. PMID- 12081249 TI - APOC at mid-point: so far so good. PMID- 12081250 TI - The challenges of community-directed treatment with ivermectin (CDTI) within the African Programme for Onchocerciasis Control (APOC). AB - The principal strategy adopted by the African Programme for Onchocerciasis Control (APOC), for the control of onchocerciasis in the 19 countries of Africa that now fall within the programme's remit, is that of community-directed treatment with ivermectin (CDTI). Halfway through its 12-year mandate, APOC has gathered enough information on the main challenges to guide its activities in Phase 2. An analysis of reports and other documents, emanating from consultants, scientists, monitors and national and project-level implementers, indicates that there are three broad categories of challenge: managerial; technical; and socio political. Under these three categories, this review identifies the most pertinent concerns that APOC must address, during Phase 2, to enhance the prospects of establishing sustainable systems for ivermectin distribution. The major challenges include: (1) maintaining timely drug-collection mechanisms; (2) integrating CDTI with existing primary-healthcare services; (3) strengthening local health infrastructure; (4) achieving and maintaining an optimal treatment coverage; (5) establishing and up-scaling community self-monitoring; (6) designing and implementing operations research locally; (7) ensuring the adequacy of community-directed distributors; (8) increasing the involvement of local non govemmental develop organizations in the programme; (9) achieving financial sustainability; (10) implementing equitable cost-recovery systems; and (11) engaging in effective advocacy. The implications of the challenges and suggestions about how they are being (or could be) addressed are also highlighted in this brief review, which should be of value to other programmes and agencies that may be contemplating the adoption of this unique strategy. PMID- 12081251 TI - Partnership and promise: evolution of the African river-blindness campaigns. AB - This article describes the evolution of the partnership, between various health and developmental agencies, that has sustained the campaign against river blindness in Africa. The international community was oblivious to the devastating public-health and socio-economic consequences of onchocerciasis until towards the end of the 1960s and the beginning of the 1970s. Then a 'Mission to West Africa', supported by the United Nations Development Programme, and a visit to the sub region by the president of the World Bank culminated, in 1974, in the inauguration of the Onchocerciasis Control Programme in West Africa (OCP). OCP was a landmark event for the World Bank as it represented its first ever direct investment in a public-health initiative. The resounding success of the OCP is a testimony to the power of the partnership which, with the advent of the Mectizan Donation Programme, was emboldened to extend the scope of its activities to encompass the remaining endemic regions of Africa outside the OCP area. The progress that has been made in consolidating the partnership is discussed in this article. The prospects of adapting the various strategies of the African Programme for Onchocerciasis Control, to entrench an integrated approach that couples strong regional co-ordination with empowerment of local communities and thereby address many other health problems, are also explored. PMID- 12081252 TI - Gender issues in the community-directed treatment with ivermectin (CDTI) of the African Programme for Onchocerciasis Control (APOC). AB - This paper reviews the issues relating to compliance and participation among the men and women of three countries within the remit of the African Programme for the Control of Onchocerciasis (APOC): Cameroon, Nigeria and Tanzania. Project monitoring data from 109 focus-group discussions, 6069 household-survey respondents and 89 interviews with ivermectin distributors were analysed to gain an insight into the attitudes and behaviours of men and women in relation to ivermectin treatment and their participation in the programme. Although there are no statistically significant gender differences in coverages for ivermectin treatment, culturally prescribed gender relationships influence the ways in which men and women express and experience treatment-related behaviours. Gender roles also affect participation in the programme. Decision-making in communities on the selection of distributors tends to follow socio-cultural hierarchies based upon patriarchy and gerontocracy. Relatively few ivermectin distributors (21%) are women. Although they receive less support than their male counterparts, the female distributors are just as willing to continue ivermectin distribution in the community, and they perform as well or better than men in this regard. The terms 'community-directed', 'community participation' and even 'compliance' obfuscate important gender differences that are inherent in the implementation of onchocerciasis control. Development of strategies that recognize these gender differences will have important implications for long-term adherence to treatment and for the overall quality and sustainability of the programme. PMID- 12081253 TI - Monitoring community-directed treatment programmes for sustainability: lessons from the African Programme for Onchocerciasis Control (APOC). AB - Community-directed treatment is a relatively new strategy that was adopted in 1997 by the African Programme for Onchocerciasis Control (APOC), for large-scale distribution of ivermectin (Mectizan). Participatory monitoring of 39 of the control projects based on community-directed treatment with ivermectin (CDTI) was undertaken from 1998-2000, with a focus on process implementation of the strategy and the predictors of sustainability. Data from 14,925 household interviews in 2314 villages, 183 complete treatment records, 382 focus-group discussions, and the results of interviews with 669 community leaders, 757 trained community directed drug distributors (CDD) and 146 health personnel (in 26 projects in four countries) were analysed. The data show that CDD dispensed ivermectin to 65.4% of the total population (71.2% of the eligible population), with no significant gender differences in coverage (P > 0.05). Treatment coverage ranged from 60.2% of the eligible subjects in Cameroon to 76.9% in Uganda. There was no significant relationship between the provision of incentives to CDD and treatment coverage (P > 0.05). The frequency of treatment refusal was highest in Cameroon (29.2%). Although most (72.1%) of the communities investigated selected their CDD on the basis of a community decision at a village meeting, only 37.9% chose their distribution period in the same way. There is clearly a need to improve communication strategies, to address the issues of absentees and refusals, to emphasise community ownership and to de-emphasise incentives for CDD. The investigation of the 'predictor indicators' of sustainability should enable APOC to understand the determinants of project performance and to initiate any appropriate changes in the programme. PMID- 12081254 TI - APOC's strategy of community-directed treatment with ivermectin (CDTI) and its potential for providing additional health services to the poorest populations. African Programme for Onchocerciasis Control. AB - Since its inauguration in 1995, the African Programme for Onchocerciasis Control (APOC) has made significant progress towards achieving its main objective: to establish sustainable community-directed treatment with ivermectin (CDTI) in onchocerciasis-endemic areas outside of the remit of the Onchocerciasis Control Programme in West Africa (OCP). In the year 2000, the programme, in partnership with governments, non-governmental organizations and the endemic communities themselves, succeeded in treating 20,298,138 individuals in 49,654 communities in 63 projects in 14 countries. Besides the distribution of ivermectin, the programme has strengthened primary healthcare (PHC) through capacity-building, mobilization of resources and empowerment of communities. The community-directed treatment approach is a model that can be adopted in developing other community based health programmes. The approach has also made it possible to bring to the poor some measure of intervention in some other healthcare programmes, such as those for malaria control, eye care, maternal and child health, nutrition and immunization. CDTI presents, at all stages of its implementation, a unique window of opportunity for promoting the functional integration of healthcare activities. For this to be done successfully and in a co-ordinated manner, adequate funding of CDTI within PHC is as important as an effective sensitization of the relevant policy-makers, healthworkers and communities on the value of integration (accompanied by appropriate training at all levels). Evaluation of the experiences in integration of health services, particularly at community level, is crucial to the success of the integration. PMID- 12081255 TI - The potential of parathyroid hormone as a therapy for osteoporosis. AB - Over the past 10 years, therapeutic optimism in osteoporosis has been fueled by the development of agents that inhibit the resorption of bone. An increase in bone density with the bisphosphonates alendronate and risedronate, for example, is associated with significant reductions in vertebral and hip fracture incidence. Although bone is clearly stronger when these agents are used, there is no conclusive evidence that they improve the microarchitectural defects that characterize the disorder. It has been known for years that parathyroid hormone (PTH) can be an anabolic agent in animals. Recently, it has been confirmed that low-dose, intermittent administration of PTH is associated with selective anabolic effects in human subjects, whereas higher doses administered continuously are associated with catabolic effects. This observation has led to an experimental design in which PTH is administered to human subjects once daily in doses that do not regularly lead to adverse events, such as hypercalcemia. Two year studies using PTH alone have been directed at postmenopausal osteoporosis (PMO) in women and at men with idiopathic osteoporosis. In both women and men, PTH leads to marked increases in bone density at the lumbar spine and also, significantly, in the hip. In PMO, PTH is associated with significant reductions in vertebral and nonvertebral fractures. When PTH is used in combination with an anti-resorptive agent, like estrogen, the markedly positive effects on bone mass are also seen in PMO and in those with glucocorticoid-induced osteoporosis. After PTH is withdrawn, but estrogen therapy is continued, the gains in bone mass are maintained. It is not known whether gains in bone mass are maintained if PTH is withdrawn without continuing therapy with an anti-resorptive agent; however, preliminary data suggest that the gains in bone mass begin to be lost. Bone biopsy samples in PMO and in men with idiopathic osteoporosis, before and after therapy with PTH, when subjected to histomorphometric analysis show marked increases in cortical thickness and improvements in indices of trabecular connectivity, thus allaying concerns that PTH may cause cortical thinning. It would appear that the increase in cortical wall thickness is due, in part, to stimulation by PTH of periosteal apposition at a rate greater than its effect to cause endocortical resorption. In all studies reported to date, PTH appears to be well tolerated. The era of anabolic therapy for osteoporosis is at hand, with PTH being the first to be tested with sufficient rigor to demonstrate its efficacy. PMID- 12081256 TI - The role of the primary care physician in diagnosis and management of osteoporosis. AB - It is estimated that 28,000,000 women in the United States have low bone mass (osteopenia) or osteoporosis. More than 1.5 million osteoporotic fractures occur annually. Because osteoporosis is asymptomatic until fracture occurs, early diagnosis requires both an awareness of risk and specific testing. The National Osteoporosis Foundation (NOF) has published guidelines for diagnosis and treatment, but these have important limitations. Bone mineral density (BMD) testing is the method of choice for diagnosis, and is more predictive of fracture risk than hypertension is of stroke or hypercholesterolemia is of myocardial infarction. Although the diagnosis of osteoporosis, as defined by the World Health Organization, is a T-score of < or =-2.5, the association between BMD and fracture risk is a continuous rather than threshold effect, and NOF guidelines suggest treating at higher T-scores, depending on risk factors. Important risk factors include age, current smoking, low body weight (<127 lbs.), maternal history of fracture, and personal history of fracture. Data from the National Osteoporosis Risk Assessment (NORA) study are presented, demonstrating the usefulness of peripheral BMD measurements in identifying postmenopausal women at risk of fracture. Several therapeutic options, including hormone replacement therapy, raloxifene, calcitonin, alendronate, and risendronate, are now available to the clinician. It can be argued that we currently have all necessary tools to eliminate osteoporosis and osteoporotic fracture. PMID- 12081257 TI - Hypothyroidism and women's health. AB - Hypothyroidism affects 4% to 10% of women, increasing with age. Symptoms, which are often nonspecific and subtle, can include: lethargy, mild weight gain, edema, cold intolerance, constipation, mental impairment, dry skin, depression, irregular menses, hoarseness, myalgias, hyperlipidemia, and bradycardia. TSH determination is usually warranted when some of these are present. Confirmation of diagnosis is based on laboratory tests, not symptoms. With Hashimoto's thyroiditis, the most common cause, the thyroid is usually firm and irregular to palpation. Screening is recommended by the American Thyroid Association, American Association of Clinical Endocrinologists and ACOG. Studies indicate that even slight hypothyroidism is associated with increased miscarriage, late fetal demise and lower IQ of offspring. Many believe all women intending conception or pregnancy should be screened. Most patients with even slightly elevated TSH should be treated, though controversy remains. Hypothyroidism is usually permanent, and treatment is life-long; postpartum thyroiditis may be transient. Levothyroxine is the standard treatment. Because of variable bioavailability of some levothyroxine preparations, designation of a specific high-quality brand is appropriate; TSH should be monitored to titrate the dose. The third-generation super-sensitive TSH is the most accurate test for both hypo- and hyperthyroidism. Because estrogen increases levels of thyroid binding globulin, women who are on HRT or OCs, or are pregnant, usually have total T4 levels above the reference range. PMID- 12081258 TI - Issues and management of herpes in pregnancy. AB - Genital herpes simplex virus (HSV) infection during pregnancy poses a significant risk to the developing fetus and newborn. In the United States, the incidence of this sexually transmitted disease has increased since 1970. Since many women of childbearing age are infected or are becoming infected, the risk of maternal transmission of this virus to the fetus or newborn is a major health concern. This review defines the stages of herpetic infection, outlines the spectrum of maternal and neonatal infection, including rates of transmission and risks, and provides management guidelines that have been validated by appropriately conducted outcome-based research. PMID- 12081259 TI - Gender issues in the management of inflammatory bowel disease and irritable bowel syndrome. AB - Crohn's disease and ulcerative colitis are two chronic inflammatory diseases involving the gastrointestinal tract. While many aspects of the management of these diseases are identical for all patients, there are some issues that are specific to women that are not necessarily part of routine care. Such gender specific issues as menses, fertility, pregnancy and menopause are often overlooked and mismanaged. Women have different psychological concerns as compared with men with regard to their self-image and impact of disease. Health care providers, regardless of their primary focus, should be aware of these differences and be familiar with general information. Irritable bowel syndrome is a prevalent condition that affects more women than men. Theories as to its underlying pathophysiology are still evolving, but what does appear clear is that women pose specific challenges to the management of this condition. PMID- 12081260 TI - Relationship between training volume and mood states in competitive swimmers during a 24-week season. AB - Mood states have been shown to be sensitive to periods of overtraining among endurance athletes. To examine the relationship between training volume and specific mood states, the Profile of Mood States was administered biweekly to 12 male and 17 female collegiate swimmers over the course of a competitive season. Pearson correlations were estimates of the relationship between mean subscores and training volume. Mean training volume for the comparison periods ranged from 3,868 to 12,500 meters/day. Mean ratings of both Anger and Vigor were significantly negatively correlated with training volume (-.58 and -.54, respectively). In addition, Fatigue was significantly correlated with training volume (r=.53), but the mean subscores for Tension, Depression, Confusion, and Global Mood were not significantly correlated with training volume. Results support previous research reporting mood disturbances with increased training volumes but do not support suggestions of an increased tendency towards higher scores on depression. PMID- 12081261 TI - Historical literacy of advanced undergraduate psychology students: pedagogical implications for courses in the history of psychology. AB - 139 junior and senior students enrolled in courses in the history of psychology at two universities were tested on the first day of class to assess general historical literacy, literacy in the history of psychology, and recognition of 51 important figures in the history of psychology. Serious deficiencies in their historical knowledge present important pedagogical implications for the teaching of the history of the discipline. PMID- 12081262 TI - Testing the modifiability of the state sport confidence inventory. AB - Completing the State Sport Confidence Inventory requires participants to compare themselves with the most confident athlete they know. Inconsistency is inherent in participants' choices that may result in unsystematic variance across such responses. This study tested the modifiability of the inventory by removing the "comparison" component from the instructions and from each of the 13 questions. Using data from 31 female figure skaters (M age=16.1 yr., SD=1.0), the original and modified versions were compared. Adequate reliabilities were found for both measures (both rs=.94), but a paired sample t test indicated significant differences between mean scores on the two versions. The total score and all of the item scores were higher for the modified version than the original, and coefficients of determination showed variance shared by the modified and original items ranged only from 9 to 59% per item. These results suggest that the modified version was not an adequate representation of the original inventory. PMID- 12081263 TI - Cortical representation of swallowing: a modified dual task paradigm. AB - It is unclear whether the cortical representation of swallowing is lateralized to the left cerebral hemisphere, right hemisphere, or bilaterally represented. As dysphagia is common in acute stroke, it is important to elucidate swallowing lateralization to facilitate earlier detection of stroke patients who may be at greater risk for dysphagia and aspiration. In this study, a modified dual task paradigm was designed to study laterality of swallowing in a group of 14 healthy, young, right-handed, male adults. The subjects were studied at baseline and with interference. Baseline conditions, performed separately, were continuous swallowing, finger tapping using the right and left index fingers, and word repetition. Interference tasks, including tapping with the right index finger, tapping with the left index finger, and word repetition, were completed with and without swallowing. Finger-tapping rate was measured, and x-ray samples of the swallowing task were taped to measure swallowing rate and volume swallowed. At baseline, the rate of tapping the right index finger was significantly faster than that of the left index finger. There was a significant decline in the tapping rates of both left and right index fingers with swallowing interference. The volume per swallow was significantly reduced during the interfering language task of silent repetition. These results offer partial support for a bilateral representation of swallowing as well as suggest an important left hemispheric contribution to swallowing. However, it cannot be concluded that the left hemisphere is more important than the right, as a comparable right hemisphere task was not studied. PMID- 12081264 TI - Karate and aggressiveness among eight-year-old boys. AB - This report is an addendum to Reynes and Lorant's study (2002). From a small karate group of 9 boys assessed for aggressiveness analysis yielded no significant variations of aggressiveness after one year of karate training but showed differences between karate and judo training which requires further investigation. PMID- 12081265 TI - Young Korean athletes' goal orientation and sources of enjoyment. AB - Feeling of enjoyment plays a critical role in sport participation and adherence. The purposes of this study were to (a) identify specific sources of enjoyment in the context of Korean youth sport and (b) examine the conceptual link between goal orientation and the sources of enjoyment in youth sport. A total of 334 middle school athletes (244 boys, 90 girls; ages 12 to 18 years), representing 17 schools in Seoul, participated. They responded to the Korean version of the Task and Ego Orientation Questionnaire in Sport and an open-ended item designed to identify sources of enjoyment. Content analysis of the responses resulted in nine enjoyment categories, including winning and competition, social recognition and rewards, health and fitness, and perceived competence. Chi squared analyses provided initial evidence for the conceptual link between goal orientation and sources of enjoyment. The athletes scoring high on task orientation more frequently reported self-referenced sources of enjoyment, e.g., health and fitness, psychological benefit, while the athletes high on ego orientation more frequently identified social recognition and rewards as their enjoyment sources. These results were discussed within the frame of goal orientation theory and cross-cultural perspectives. PMID- 12081266 TI - Active inhibition of stuttering results in pseudofluency: a reply to Craig. AB - Craig discussed fluency outcomes following stuttering therapy that involved retraining the speech system, on the assumption that the speech end product is truly fluent. As previously outlined by Dayalu and Kalinowski, we strongly disagree with the notion that behavioral paradigms can ever result in automatic, long lasting natural sounding fluent speech. Fluent speech is within the grasp of one who stutters as seen in the effects of choral speech and derivatives such as delayed auditory feedback. PMID- 12081267 TI - No sex differences in contrast sensitivity and reaction time to spatial frequency. AB - This study investigated the possibility of sex differences in spatial frequency processing by measuring contrast sensitivity and reaction time to spatial frequency in the same 20 men and 20 women. This is the first study to investigate sex differences in reaction time to spatial frequency and the first to study sex differences in contrast sensitivity and reaction time within the same participants. No sex differences were found in either contrast sensitivity or reaction time measures, suggesting that women and men process spatial frequency information similarly. PMID- 12081268 TI - Relationships between performance toward accomplishment and self-efficacy in amateur boxing. AB - Combined ratings of magnitude and strength of self-efficacy by 59 20-yr.-old amateur boxers in a multiple regression were predicted only by Perceived Performance, i.e., their working toward successful performance. PMID- 12081269 TI - Judo training and aggression: comment on Reynes and Lorant. AB - The Reynes and Lorant longitudinal study suggests evidence of an increase in aggression in 8-year-old male judo students after one year of traditional judo training. However, several methodological problems make these results difficult to interpret. Nonetheless, the authors make a valuable contribution to the area of the effects of traditional martial arts training and aggression. PMID- 12081270 TI - Right-left discrimination in younger and older children measured with two tests containing stimuli on different abstraction levels. AB - Two paper-and-pen tests, consisting of line drawings of human figures, viewed from the back, the front, or randomly alternating between back and front drawings, were used to study right-left discrimination in younger and older children. One test, called the Abstract test, had just a circle to indicate heads of the figures. The other test, called the Concrete test, showed facial characteristics and hair in the head circle. The sample consisted of 280 younger and older children, ages 7-8 and 12-13 years, respectively. The main findings were that participants who responded to the Concrete test solved more items correctly compared to those using the old figures in the Abstract test. The error scores did not differ between the tests. The older children solved more items correctly than the younger children. The older children also showed a significant decrease in correct scores on the subtest with the alternating views subtest compared with the other subtests. Their error scores did not differ across the subtests. The younger children solved a similar number of correct items on all subtests; however, they made significantly more errors on the subtest using the front view than on the other two subtests. The results are discussed in relation to theories of hemispheric lateralization, brain development, and cognitive development. PMID- 12081271 TI - Psychophysiological comparison of self-efficacy and resting heart rate. AB - This study was used to assess the premise that lower resting heart rates should be associated with higher scores on self-efficacy for exercisers. A total of 64 participants, 29 men and 35 women ranging in age from 18 to 38 years old (M = 23.22, SD = 4.19), currently participating in an exercise program at a fitness center, were assessed using the Exercise Specific Self-efficacy Scale to measure general individual self-efficacy, along with obtaining resting heart rates. Participants were separated into three different groups based on individual resting heart-rate values. Results for a 2 (sex) by 3 (heart rate) two-way analysis of variance indicated a significant difference in self-efficacy scores (F(2,58) = 3.24, p<.05) among the three groups. No interaction or main effect for sex was found. PMID- 12081272 TI - Ratings of perceived exertion during low- and high-intensity resistance exercise by young adults. AB - Ratings of perceived exertion (RPE) are commonly used to monitor the intensity of aerobic exercise. Whether ratings of perceived exertion can be used similarly during resistance exercise is unclear. To examine this question, perceived exertion was measured at 30% and 90% of the one-repetition maximum (1-RM), while holding work constant between intensities. Ratings for the active muscles and for the overall body were examined during both intensities. 10 male (age=23.2 +/- 3.6 yr.) and nine female (age=21.8 +/- 2.7 yr.) volunteers underwent a one-repetition maximum procedure for each of the following exercises: bench press, leg press, latissimus pull down, triceps press, biceps curl, shoulder press, and calf raise. All subjects then completed two experimental trials on separate days. The high intensity trial consisted of one set of five repetitions at 90% of the one repetition maximum. The low-intensity trial consisted of one set of 15 repetitions at 30% of the one-repetition maximum. Active muscle and overall body ratings of perceived exertion were obtained immediately at termination of each of the seven exercises at both intensities. A two-factor (RPE x Intensity) repeated measures analysis of variance was performed separately for each exercise. Both active muscle and overall body ratings of perceived exertion were higher (p<.01) for the high-intensity trial than for the low-intensity trial. Active muscle ratings were higher (p<.01) than overall body ratings for all exercises. Ratings of perceived exertion during resistance exercise are related to intensity of the resistance exercise (percentage of the one-repetition maximum). This information suggests that ratings of perceived exertion can provide information regarding the intensity of resistance exercise. Furthermore, sensations of exertion in the active muscles during resistance exercise are greater than sensations for the overall body. PMID- 12081273 TI - Mood changes following exercise. AB - 26 participants completed a mood measure to assess anger, confusion, depression, fatigue, tension, and vigor immediately before and immediately after two exercise sessions. Analysis showed significant mood enhancement for each exercise session. Follow-up univariate results indicated that Depressed mood scores were reduced significantly and Fatigue scores increased significantly following the first exercise session. Scores after the second exercise session indicated that Depressed mood decreased significantly. There was no interaction. Results lend support for the notion that exercise reduces depressed mood scores. It is suggested that researchers should consider the mechanisms that produce changes in mood following exercise. PMID- 12081274 TI - Effects of differing durations and intensities of cardiovascular exercise on aversion and feeling states in new women exercisers. AB - 20 previously sedentary women completed three assigned cardiovascular exercise protocols, each of varying lengths (15 to 25 min.) and intensities. No statistically significant difference was found on measures of pre- to postexercise changes in feeling states, or aversion, between protocols. Findings suggest that, contrary to previous research, longer exercise sessions may not always be less acceptable to participants than shorter ones, especially when a "cool down" period is added at the end. Assigned exercise durations and intensities may, however, be less likely to be associated with positive postexercise changes in feelings than self-selected amounts. PMID- 12081275 TI - Debilitative and facilitative perceptions of trait anxiety among students in a college golf class. AB - This study examined the debilitative and facilitative perceptions of trait anxiety among 94 students enrolled in a college golf class. Analyses indicated no differences in performance between those classified as debilitating interpreters, i.e., those with negative perceptions of trait anxiety, versus those classified as facilitating interpreters, i.e., those with positive perceptions of trait anxiety. Also, no differences were found in perceived intensity and direction of trait anxiety as a function of performance. Results differ from previous research on direction of trait anxiety among elite and high level performers so direction of perceptions may be correlated with competition or skill of the performer. PMID- 12081277 TI - Rehearsal of task variations and contextual interference effect in a field setting. AB - The present study showed a consistent pattern with a typical contextual interference effect in motor skill learning (N=38 adults). Individuals trained under the high contextual interference condition may receive greater learning benefits than those under the low contextual interference condition during the rehearsal of task variations of a single motor skill in a field setting. PMID- 12081276 TI - Sex-role stereotyping for selected sport and physical activities across age groups. AB - This investigation was designed to assess sex-role stereotyping across age groups. Participants (N=668) were girls and boys, students from Grades 3, 5, 8, and 10 at local public schools. All participants completed the Sport and Physical Activities Questionnaire on which were displayed pictures of 31 sport and physical activities. Participants were instructed to designate each activity as a boys' activity, a girls' activity, or a boys' and girls' activity. Chi-square analysis showed age-related differences in distribution of stereotyping of the activities. Over age groups there were more discrepancies between boys' and girls' ratings of activities as sex-specific. These findings suggest that sex role stereotyping of sports and physical activities becomes more predominant across age groups. PMID- 12081278 TI - Changes in sleep patterns during simulated nitrox saturation diving to 20 and 30 meters. AB - This study assessed the effect of a nitrogen-oxygen (N2-02) saturation diving environment on nocturnal sleep and the differences in sleep on diving depth. We measured and recorded the standard polysomnograph and heart rate for a total of 459 nights on 27 divers who performed the N2-O2 saturation simulation dive an 20- and 30-m equivalent depths as well as their subjective feelings of fatigue. From the last half of the period on the bottom to the postdive period of both saturation diving depths, our main findings included a prolongation of sleep latency, shortened total sleep time, a decreased sleep efficiency index, and increased feelings of fatigue. During the bottom period and decompression period, the total number of awakenings and the awakenings from rapid eyes movement (REM) sleep increased. This was slightly greater at the 30-m equivalent depth. From these facts, we assumed that the deterioration of sleep at the 20- and 30-m nitrox saturation dives was predominantly relative to being restricted in a closed environment for a long time and the lack of physical activity of the divers. No recovery of sleep was observed in the postdive period. A difference in the effects on sleep by the dive depth was not found. PMID- 12081279 TI - College-age males' ability to produce the acoustic properties of an aging voice.. AB - The purpose of this study was to examine college-age males' ability to produce the acoustic properties of the normally aging voice when reading. The 17 subjects (M age=21.13 yr., SD=1.0) selected for this study were undergraduates who were placed into a single group. The procedure involved recording the subjects while reading The Rainbow Passage aloud. The first reading was in the subject's natural speaking voice. During the second reading, the reader imitated the voice of a normally aging 70-yr.-old man. Fundamental frequency and temporal measures were analyzed for each voice sample. Mean scores for each measure were compared for the natural speaking-voice production and the production when imitating the voice of a normally aging 70-yr.-old man. Analysis showed that temporal measures appear to have the most significant influence on subjects' production when imitating the normally aging voice as seen in the overall increase in all temporal measures. PMID- 12081280 TI - Fluency outcomes following treatment for those who stutter. AB - The efficacy of treatment therapies for stuttering has been questioned recently by Dayalu and Kalinowski. Aspects put forward that raise doubts about stuttering treatment efficacy included posttreatment unnatural speech and high relapse rates. However, this paper examines these claims and puts forward counterclaims suggesting that at least some stuttering therapies have been shown by the best available research to be efficacious. PMID- 12081281 TI - Self-reported anxiety of adults who do and do not stutter. AB - This is a preliminary investigation of the differences in self-reported anxiety of people who do and do not stutter. Participants and experimental procedures were the same as those used by Gabel, Colcord, and Petrosino in 2001. 10 people who stuttered, all of whom had experience with stuttering therapy, were matched with 10 participants who did not stutter. Each participant in both groups was engaged in a session to evaluate speech in a format similar to a professional diagnostic session for stuttering. Each participant was stopped and asked to rate his anxiety during specific moments during the session. This procedure is similar to in vivo cognitive assessment used in cognitive-behavioral treatments and research. The participants' reports were taken during a baseline period, during a period in which they were thinking about their speech, and during three different speaking tasks. The anxiety reported by the group of people who stuttered was compared to the anxiety reported by the group of individuals who did not stutter. Statistical analyses indicated a significant main effect for the group who stuttered as they reported more anxiety during the entire session. There was no main effect for condition. The interaction of group by condition fell short of significance. Possible explanations and suggestions for research were discussed. PMID- 12081282 TI - Predictive effectiveness of mood measures. AB - The present study compared the predictive effectiveness of mood states and the Positive and Negative Affect Schedule models of mood. 38 junior high school students completed the Brunel University Mood Scale to assess anger, confusion, depression, fatigue, tension, and vigor and the PANAS 5 min. before a fitness test. Performance was self-referenced by comparing fitness test scores with a previous best score assessed four weeks earlier. Participants who equaled or bettered their previous best score were classified as Successful. Those who did not match their previous score were classified as Unsuccessful. Discriminant function analysis indicated that 31 participants (81.6%) could be correctly classified as Successful or Unsuccessful based on preperformance Brunel Mood scores. Discriminant function coefficients indicated that Confusion, Fatigue, and Vigor contributed significantly to the discrimination. For the Positive and Negative Affect Schedule, discriminant function analysis indicated that 26 participants (68.4%) could be correctly classified as Successful or Unsuccessful, with Negative Affect and Positive Affect contributing significantly to the discrimination. Summarizing negative mood into a single factor may lead to a substantial loss of information. Research on mood and performance relationships should use the mood dimensions assessed in the Brunel University Mood Scale rather than the Positive and Negative Affect Schedule. PMID- 12081283 TI - Note on learners' control of the frequency of model presentation during skill acquisition. AB - Learners (30 men, 15 women in three groups), given an opportunity to control the frequency with which they viewed a model during initial practice of the badminton long serve, acquired and retained a level of movement form that was equivalent to learners who viewed the model 100% of the time and significantly superior to learners who never viewed the model. PMID- 12081284 TI - Relation of burnout with lack of time for being with significant others, role conflict, cohesion, and self-confidence among Norwegian Olympic athletes. AB - We studied the relationship of burnout with environmental and personal characteristics such as lack of time for being with significant others outside sport, cohesion in training groups, role conflict, and self-confidence among 136 elite athletes. Analysis indicated that the mean Burnout scores were in the low range. Cohesion in training groups and Self-confidence were negatively associated with Burnout, whereas Lack of time to be with significant others and Role conflict were positively ssociated with Burnout scores. Results are in accordance with Coakley's 1992 conception of burnout explained as a social problem, Kahn's 1978 hypothesis that role conflict is positively correlated with stress and burnout, and Smith's 1986 hypothesis that personality factors such as self confidence should be associated with cognitive appraisal of situational demands related to burnout. PMID- 12081285 TI - Performance satisfaction and postcompetition mood among runners: moderating effects of depression. AB - This study examined the influence of postcompetition depressed mood on the intensity of other mood dimensions assessed by the Profile of Mood States and relationships between mood and satisfaction with performance. 195 distance runners completed the Brunel Mood Scale which is a 24-item measure assessing anger, confusion, depression, fatigue, tension, and vigor, and a Performance Satisfaction questionnaire immediately after running a 10-mi. race. To examine the proposed moderating effect of depression, participants were separated into a No-depression group of 133 and a Depressed Mood group of 62. Multivariate analysis of variance indicated that the Depressed Mood group reported significantly higher scores on Anger, Contusion, Fatigue, and Tension and lower Vigor scores than the No-depression group t ratios indicated that poorer performance was associated with higher Depression scores. Standard multiple regression to predict Performance Satisfaction scores from postcompetition scores on Anger, Confusion, Fatigue, Tension, and Vigor in the No-depression group indicated that only 1% (Adj. R2=.01, p>.05) of the variance was explained. By contrast, in the Depressed Mood group, the same mood dimensions predicted 27.5% of the variance in Performance Satisfaction scores (Adj. R2=.275, p<.01) with Anger (Beta=.63, p<.001), and Confusion (Beta=.44, p<.05) being significant predictors. Present findings support the notion that depressed mood is related to the intensity of other mood dimensions assessed by the Profile of Mood States and moderates relationships between those scores and performance satisfaction. PMID- 12081286 TI - Determinants for stair climbing by elderly from muscle morphology. AB - The relationship between muscle cross-sectional area (CSA) of the hip joint and lower extremities and speed of stair ascent/descent was investigated in a sample of 127 elderly women (ages: 60-70 years old). The participants performed ascent and descent of stairs with 12 steps for which the speeds for ascent and descent separately from video film were calculated. The ascent speed significantly correlated with CSAs of knee extensor (r=.21, p<.05) and psoas major muscle (r=.25, p<.01), which were assessed by magnetic resonance imaging. The descent speed also correlated with CSAs of the same muscle groups (with knee extensor: r=.20, p<.05, with m. psoas major: r=.18, p<.05). The rest of the muscle groups were not related to the ascent/descent speeds of the step. These results suggested that the muscle mass of the knee extensor muscle group and also the psoas major muscle could be a factor in potential capacity in stair ascent/descent by humans. PMID- 12081287 TI - Reactivity and event-related potentials in attentional tests: effect of training. AB - To study the effects of training on reactivity and event-related potentials a complex attentional shifting test involving reaction time was administered (Test 1) to 24 healthy, young students. After five days, 12 subjects were tested with the same procedure (Test 2) without training (Untrained Subjects) while 12 repeated the test at the fifth day after four days of training (Trained Subjects). During Tests 1 and 2, event-related potentials were recorded by electroencephalogram. The task consisted of each subject responding to a stimulus of a letter appearing in the centre of a geometric figure on the screen of a computer monitor. In the prestimulus period black points were drawn and crowded randomly into a zone of the screen. The geometric figure and the letter were shown in the centre of the crowding. There were two letters and four geometric figures randomly combined in different ways. The subject had to press different keys of the computer keyboard when specific combinations appeared. The averaged event-related potentials were characterized by a negative wave with a close relationship to selective attention before the onset of the stimulus of a geometric figure followed by letters. After the stimulus onset, a P3 complex was recorded. Trained subjects were no different from untrained subjects in Test 1, while in Test 2 they had a shorter reaction time, an earlier peak of the selective attention related wave and P3, and a higher amplitude for the P3 complex. These measures and the correlations between them can be considered an index of the training effect. Thus, these tests could be used for evaluation of the attentional style and its modification with training. PMID- 12081288 TI - Effect of added mass on human unipedal hopping. AB - Although hopping is considered a children's activity, it can be used to provide insight into the neuromuscular and biomechanical performance of adults. This study investigated whether mass added during unipedal hopping altered the vertical stiffness, hopping period, and angular kinematics of the lower extremity of adults. Measures of two-dimensional kinematics and vertical force were made from 10 healthy men during hopping at a preferred period under three conditions: Body Mass, Body Mass+10%, and Body Mass+20%. Adding mass significantly increased hopping period and hip flexion without significantly affecting vertical stiffness, ankle dorsiflexion, or knee flexion. Overall, the findings agreed with predictions based on a simple-mass spring model. The results indicate unique kinetic and kinematic responses to increased mass during hopping may have potential application in neuromuscular assessment and training for the lower extremities. PMID- 12081289 TI - Phantom smelling. AB - A case of phantom smelling (phantosmia) is described in a 28-yr.-old man who developed permanent bilateral anosmia after a serious injury to olfaction-related brain structures at the age of 25 years. The findings indicate that, even years after loss of input from olfactory receptors, the neural representation of olfactory perception can still recreate olfactory sensations without any conscious recall of them. This indicates that the neural representation of olfactory sensations remains functional and implies that neuronal activity in the olfactory organ or in other brain structures gives rise to olfactory experiences perceived as originating from the perception of original odor substances. The report suggests the intriguing possibility that the olfactory perception is not a passive process that merely reflects its normal input from the olfactory system but is continuously generated by a neural representation in the olfactory organ or in other olfaction-related brain structures, based on both genetic and sensory determinants. To the author's knowledge this is the first reported case of its kind. PMID- 12081290 TI - Measurement of ventilatory threshold by respiratory frequency. AB - This study was conducted to assess whether respiratory frequency can be used as a valid parameter for estimating ventilatory threshold and for examining differences in exercise modes such as a cycle ergometer and a treadmill. 24 men and 12 women performed an incremental exercise test to exhaustion on a cycle ergometer and on a treadmill. Oxygen uptake, carbon dioxide output, pulmonary ventilation, ventilatory frequency, and heart rate were measured continuously every 30 sec. during the test. Three different and independent reviewers detected the ventilatory threshold point and break point of respiratory rate, which were then compared. Analysis indicated that (1) ventilatory threshold was well correlated with break point of respiratory rate for both cycle (r=.88, p<.001) and treadmill exercise (r=.96, p<.001). However, on the average, ventilatory threshold was only 71% (cycle) or 88% (treadmill) of break point of respiratory rate. (2) The regression equation for treadmill exercise was more accurate than that for cycling, but the detected data samples were smaller. The break point of respiratory rate was more easily detected for the cycle ergometer test 33 of 36 subjects) than for the treadmill test (only 15 of 36). The cycle ergometer test identified the break point of respiratory rate more easily than did the treadmill test. (3) There was an association between physical fitness and whether the break point of respiratory rate was detectable, and the more fit the subject (above average), the more likely the break point was to be undetected. Our study demonstrates that the break point of respiratory rate is closely associated with ventilatory threshold and that the cycle ergometer test is more conducive than the treadmill test to the detectability of break point of respiratory rate. PMID- 12081291 TI - Relationship between phonemic and lingual awareness. AB - The purpose of this preliminary investigation was to assess the relationship between knowledge of speech sounds (phonemes) and lingual awareness in normal adult speakers of English. The study also examined subjects' descriptions of lingual contact. 36 subjects (M age=19.6, SD=.71) who were enrolled in an undergraduate phonetics class participated. All passed a speech and hearing screening and reported having received no prior speech therapy or classes in articulation, phonetics, or speech science. During the first class meeting, the subjects were given two tests. The measure of phonemic awareness consisted of (a) judging which two (out of three) printed words began (or ended) with the same phoneme, (b) counting, substituting, reversing phonemes in words, and (c) indicating primary stress in two-syllable words. The lingual awareness test consisted of subjects imitating a syllable, then responding to a multiple-choice question regarding (a) tongue position (front to back), (b) tongue height (high to low), (c) contact with the teeth, and (d) contact with other oral cavity structures. Subjects were requested to imitate the syllable prior to answering each question, e.g., "Say tuh. Did you feel your tongue in the front, middle, or back of your mouth?" Seven English phonemes (t, k, s, sh, r, l, and voiceless th) were presented in a consonant-vowel syllable with the central vowel "uh." Subjects were aided by a line drawing of the oral cavity. A significant correlation of .53 was found between the two tests, suggesting that individuals who possess greater awareness of speech sounds tend to exhibit heightened lingual awareness. Sound-symbol knowledge was the best predictor of lingual awareness. Subjects had the most difficulty describing lingual contact for phonemes that are often troublesome for children to articulate (sh, r, l, and s). PMID- 12081292 TI - Urine alarms and prompts for fostering daytime urinary continence in a student with multiple disabilities: a replication study. AB - The study evaluated a treatment approach based on urine-alarm signals and staff prompts to foster daytime urinary continence of a student with multiple disabilities. Reinforcement was used for urination in the toilet. The study alternated two baseline with two treatment phases. A 3-mo. follow-up also occurred. The student became almost totally free from large urinary accidents, replacing them with self-initiated toileting and small accidents (wetting only a disposable tissue), and retained his improvement over time. Implications of the findings were discussed. PMID- 12081293 TI - Training and assessing complex decision-making in a virtual environment. AB - The current report describes a procedure for training and assessing complex decision-making in a virtual environment. Focusing on small unit leaders, 7 experienced and 7 inexperienced Army platoon leaders performed missions in a combat simulator, where they were required to direct the activities of 3 subordinate leaders and computer-generated forces in 4 different operations in a virtual urban setting. Objective and subjective assessments of the training value of the simulations showed that both experienced an inexperienced platoon leaders improved their decision-making across the four missions, and both groups rated this "virtual environment" training procedure as useful and positive. Lessons learned and implications for designing similar training protocols for other domains are discussed. PMID- 12081294 TI - Application of Fitts' law to individuals with cerebral palsy. AB - Cerebral palsy is a condition that results in motor abnormalities as a direct consequence of injury to the developing brain. Fitts' law, which describes a speed-accuracy tradeoff in visually guided movements, has been shown to characterize the motor behavior of normal subjects during aiming tasks. To assess whether Fitts' law can also describe the aimed movements of persons with cerebral palsy, eight cerebral palsied adults participated in an aimed movement study. 12 targets were used with Indices of Difficulty ranging from 2.19 to 6.00 bits. The impact of Gan and Hoffmann's 1988 ballistic movement factor, square root(A) , and Fitts' 1954 Index of Difficulty on subject's movement and reaction times was examined using multivariate linear models. The analysis of the full data set yielded a significant effect of square root(A) on movement times and no significant adherence to Fitts' law. However, high error rates that could be the result of oculomotor problems among the subject group were noted, and the method of handling errors had a large effect on the results. Tracking eye position during a Fitts' law task would provide information regarding the effect of oculomotor difficulties on aiming tasks in the cerebral palsied subject group. PMID- 12081295 TI - Closure principle: relations across domains. AB - The present study examined a possible relationship between perceptual closure and social need for closure. These closure abilities were not correlated across domains of application. PMID- 12081296 TI - Development of an apparatus to estimate coordinated exertion of force. AB - The purpose of this study was to develop a practical apparatus for estimation of the coordinated exertion of force and to develop methods for display, test duration, and sampling interval. A total of 84 healthy male college students were required to match their grip strength to a changing demand value appearing on the display of a personal computer (using two descriptions in the form of a bar chart and wave form). The variables were the maximal difference and the total sum of the differences between demanded and produced grip strength over time. In estimating coordinated exertion of force, an actual force (kgf) scale was more effective than a relative force (%) scale. Reliability tests indicated that a 30 sec. test was effective. Both bar chart and wave form displays were effective. PMID- 12081297 TI - Velocity, occlusion, and sex of subjects in coincidence of anticipation. AB - The purpose of this study was to assess effects of velocity, occlusion distance, and sex on coincidence of anticipation. The Bassin anticipation timer was used to test 25 men and 25 women when velocity was .69 m/sec. and 1.38 m/sec. or "slow" and "fast", and occlusion distances of zero, 1.06 m and 2.12 m were "zero," "small," and "large" on a 4.25-m length of track. There were six test conditions. Analysis showed that increased occlusion distances produced increased absolute, constant, and variable error scores. A significant interaction of velocity and occlusion appeared for all error measures. With an increase in occlusion distance at the slow velocity constant error increased in the positive direction or early response while at the fast velocity a similar increase in constant errors was seen but in the negative direction or late response. Under zero occlusion the expectation that performance at the fast velocity would be more accurate and show less variability than at the slow velocity was not supported. Also, sex did not significantly affect anticipation time in any condition or on any error measures. PMID- 12081298 TI - Long-term predictive validity of the Miller assessment for preschoolers. AB - The purpose of this study was to examine the predictive validity of the Miller Assessment for Preschoolers in a sample of children in Israel. 30 children (M age 7 yr., 8 mo.) were selected from a group of typical children registered in 4 Mother and Child Health Care Centers in central Israel; they were originally tested as preschoolers (M age 3 yr., 10 mo.) on this procedure. They were retested after an interval of 5 to 7 yr. on academically related tests. Correlations between the Total scores and most of the follow-up test scores indicate significant correspondence (.41-.70). Regression analysis indicated that visuomotor integration and handwriting accounted for 58% of the variance. These results contribute to the empirical evidence regarding the predictive abilities of this procedure and indicate its validity cross-culturally over an extended follow-up interval of 5 to 7 yr. PMID- 12081299 TI - Remote viewing with the artist Ingo Swann: neuropsychological profile, electroencephalographic correlates, magnetic resonance imaging (MRI), and possible mechanisms. AB - In the present study, the artist Ingo Swann, who helped develop the process of remote viewing (awareness of distant objects or places without employing normal senses), was exposed during a single setting of 30 min. to specific patterns of circumcerebral magnetic fields that significantly altered his subjective experiences. Several times during subsequent days, he was asked to sit in a quiet chamber and to sketch and to describe verbally distant stimuli (pictures or places) beyond his normal senses. The proportions of unusual 7-Hz spike and slow wave activity over the occipital lobes per trial were moderately correlated (rho=.50) with the ratings of accuracy between these distal, hidden stimuli and his responses. A neuropsychological assessment and Magnetic Resonance Imaging indicated a different structural and functional organization within the parieto occipital region of the subject's right hemisphere from organizations typically noted. The results suggest that this type of paranormal phenomenon, often dismissed as methodological artifact or accepted as proofs of spiritual existence, is correlated with neurophysiological processes and physical events. Remote viewing may be enhanced by complex experimentally generated magnetic fields designed to interact with the neuromagnetic "binding factor" of consciousness. PMID- 12081301 TI - Perceptual judgment of voice pitch during pitch-matching tasks. AB - This study investigated the perceptual judgment of voice pitch. 24 individuals were assigned to two groups to assess whether there is a difference in perceptual judgment of voice pitch during pitch-matching tasks. Group I, Naive listeners, had no previous experience in anatomy, physiology, or voice pitch-evaluation methods. Group II, Experienced listeners, were master's level speech-language pathologists having completed academic training in evaluation of voice. Both groups listened to identical stimuli, which required matching audiotaped voice pitch samples of a male and female voice to a note on an electronic keyboard. The experiment included two tasks. The first task assessed pitch range, which required matching of the lowest and highest voice pitch of both a male and female speaker singing /a/ to a note on a keyboard. The second task assessed habitual pitch, which required matching of the voice pitch of a word spoken by a male and female speaker to a note on a keyboard. A one-way analysis of variance indicated a significant difference between groups occurred for only one of four conditions measured, perceptual judgment of the female pitch range. No differences between groups were found in the perceptual judgments of the male pitch range or during perceptual judgment of the female or male habitual pitch, suggesting that the skill possessed by speech-language pathology students is no different from that of inexperienced listeners. PMID- 12081300 TI - Two ways related to performance in elite sport: the path of self-confidence and competitive anxiety and the path of group cohesion and group goal-clarity. AB - A model tested among 136 Norwegian Olympic-level athletes yielded two paths related to performance. The first path indicated that self-confidence, modeled as an antecedent of competitive anxiety, is negatively correlated with anxiety. Competitive anxiety in turn is negatively correlated with performance. The second path indicated that group cohesion is positively correlated with group goal clarity, which in turn is positively correlated with performance. Competitive anxiety mediates the relation between self-confidence and performance, whereas group goal-clarity mediates the relation between group cohesion and performance. Results from multiple regression analyses supported the model in the total sample and among individual sport athletes organized in training groups (n = 100). Among team sport athletes (n = 36), personality and group measures are more strongly intercorrelated than among individual sport athletes, and the relation with performance is more complex for the former group. The interaction of self confidence and competitive anxiety is related to performance among team sport athletes. PMID- 12081302 TI - Arguments against the cognitive dysmetria hypothesis of schizophrenia. AB - Recently, the 'cognitive dysmetria' theory for schizophrenia has been formulated. According to this theory, a primary neurocognitive dysfunction is the core of schizophrenia and underlies symptom formation. The suggested perceptual fragmentation of external stimuli and inability to connect such perceptions with internal schemata is suggested to lead to positive symptoms, while defensive self restriction and the exhaustion of the mental apparatus lead to negative symptomatology. Objections to this theory include observations (i) that patients with dominant positive symptoms, e.g., delusions, hallucinations, manifest better neurocognitive function and (ii) that typically antipsychotics significantly reduce positive symptoms and thus improve both the clinical picture and the functioning (to the extent it is reduced with positive symptoms) of the patients, yet have little or no effect on negative, e.g., loss of volition, emotional blunting, and neurocognitive symptomatology, e.g., attentional and memory deficit. The literature suggests that neurocognitive symptoms group independently of other symptomatology. It is suggested that there is currently more evidence against than in favor of the 'cognitive dysmetria' theory. PMID- 12081303 TI - Nicotine deprivation effects on the dissociated components of simple reaction time. AB - The effect of smoking abstinence on the dissociated components of a simple reaction-time response was investigated. 7 subjects completed two experimental sessions conducted one week apart. Each experimental session was comprised of two tests. The first test consisted of abstinent baseline measures, while the second test was administered after smoking either an investigator-supplied nicotinised or denicotinised cigarette. A within-subjects design was utilized which permitted each subject to be their own control. Interestingly, the smoking of either experimenter-controlled cigarette (nicotinised or denicotinised) reliably decreased the subjects' desire to smoke. More importantly, however, was that only the nicotinised cigarette had any influence on simple reaction time. Moreover, this decrease in reaction time was isolated within the premotor or cognitive processes. In fact, there was no evidence that the smoking of a nicotinised cigarette had any effect on the motor-time component of this task. These findings are relevant for two reasons, (1) performance decrements due to smoking abstinence were independent of the urge or craving to smoke and (2) nicotine affects cognitive and motor processes of a simple reaction-time task differently. PMID- 12081304 TI - Self-rating inventory for posttraumatic stress disorder: review of the psychometric properties of a new brief Dutch screening instrument. AB - The Self-rating Inventory for Posttraumatic Stress Disorder of 22 items was developed for use with populations without identified traumatic experiences. The inventory has been used extensively in survey research in The Netherlands. This paper examines the psychometric properties. In four different groups (trauma and psychiatric patients, elderly Dutch subjects, former peacekeepers, and medical students) internal consistency, test-retest reliability, concurrent and discriminant validity, and sensitivity and specificity are analyzed. The inventory showed good internal consistency, test-retest reliability, concurrent and discriminant validity, and high sensitivity and specificity. It appears to be valuable for survey research on posttraumatic stress in nonselected populations. As a screening device, high sensitivity for PTSD symptoms is evident even when the traumatic event has not been defined. PMID- 12081305 TI - Fertility of dairy cows in Northern Ireland. AB - A comprehensive database was established on the milk production and reproductive performance of dairy cows in 19 selected herds in Northern Ireland, varying in size, management system and genetic merit. Data were obtained for 2471 cows, 1775 of which calved in a second year, and 693 were culled from the herd for specific reasons. The estimated mean rate of heat detection (assessed by the interheat interval during the main breeding season) in all the herds was 71 per cent, with a range from 53 to 92 per cent The average conception rate to first insemination was 37.1 per cent (range 21 to 66 per cent). The average calving interval for the retained cows was 407.2 days (range 359 to 448 days). Twenty-eight per cent of the cows that calved were culled, with infertility being the largest single reason (26.8 per cent of the cows culled). There were major differences in reproductive performance between the herds, but heat detection rate, conception rate and calving interval did not appear to be affeded by a herd's genetic merit. The herds with shorter calving intervals were characterised by better heat detection efficiency (83 v 61 per cent, P<0.01), a shorter interval from calving to first insemination (74 v 97 days; P<0.05), a higher conception rate to first insemination (45 v 34 per cent, P>0.10) and a lower removal rate (23 v 37 per cent, P<0.01). Furthermore, the cows in these herds had lower body condition scores (BCS) in the dry period (3.0 v 3.3; P<0.05) but lost less body condition in early lactation (0.3 v 0.6 BCS units, P<0.05). These results show that dairy herd fertility in Northern Ireland is generally low and similar to that previously reported for England and the USA, but that in some herds changes in herd management practices improved the cows' fertility. PMID- 12081306 TI - Evaluation of the efficacy of oral lufenuron combined with topical enilconazole for the management of dermatophytosis in catteries. AB - The efficacy of oral lufenuron, a chitin synthetase inhibitor, combined with topical enilconazole, was evaluated for the management of Microsporum canis infection in 100 cats housed in two catteries in France. The cats were treated with weekly rinses with enilconazole (0.2 per cent) for four weeks and, in each cattery, one group (A) was also treated with micronised griseofulvin (25 mg/kg administered orally twice a day for five weeks) and a second group (B) was treated with 60 mg/kg lufenuron administered orally once on day 0 and again after 30 days. All the cats were examined individually for cutaneous lesions and mycological cultures were made when the treatment began and after 15, 30, 60 and 90 days. In the first cattery, the cats' clinical scores after 30 and 60 days were significantly lower in group B than in group A. In both catteries and both treatment groups, the mean number of fungal colonies decreased rapidly during the first 15 days of treatment, remained stable for the following 45 days but increased from day 60 to the end of the experiment on day 90. PMID- 12081307 TI - Phenobarbitone concentrations in the hair, saliva and plasma of eight epileptic dogs. AB - Samples of plasma, saliva and hair were collected from eight dogs receiving phenobarbitone for idiopathic epilepsy. The concentrations of phenobarbitone in hair and plasma were correlated with the daily dose rate, and the concentrations in hair were also correlated with the concentration in plasma, the dose rate of the drug over the preceding six months and the ratio of the six-month dose to body surface area. The concentration of phenobarbitone in saliva was not correlated with either the concentration in plasma or the dose rate of the drug. PMID- 12081308 TI - Foot-and-mouth disease virus infection of sheep: implications for diagnosis and control. PMID- 12081309 TI - Cutaneous candidiasis in a dog caused by Candida guilliermondii. PMID- 12081310 TI - Competition Commission inquiry into the supply of POMs. PMID- 12081311 TI - Large animal parasitology. PMID- 12081312 TI - Euthanasia of NGRC-registered greyhounds. PMID- 12081313 TI - Cost of animal disease epidemics. PMID- 12081314 TI - Centre for Tropical Veterinary Medicine. PMID- 12081315 TI - Binding affinities of commonly employed sensitizers of viral inactivation. AB - Methylene blue (MB), riboflavin (RB) and psoralen sensitizers (4' aminomethyl 4,5',8-trimethylpsoralen [AMT] and derivatives) are under study as sensitizers of viral inactivation of blood products such as plasma proteins, platelets and red cells, all of which lack genomic nucleic acid. To predict where these sensitizers accumulate in viruses and in cells, their relative affinities for calf thymus DNA, neutral and negatively charged phospholipids and albumin were determined by dialysis. MB has a strong affinity for nucleic acid and negatively charged phospholipid, but little affinity for albumin or neutral phospholipid. RB has modest affinity for nucleic acid and little affinity for albumin or either phospholipid. AMT has substantial affinity for nucleic acid, neutral and negatively charged phospholipids and albumin. Neither AMT nor RB binds to poly G, although MB has some affinity for this polymer. Evidence of association of RB with guanosine monophosphate, adenosine monophosphate and tryptophan methylester hydrochloride in PBS buffer in the presence and absence of formamide was obtained from nonlinear Stern-Volmer plots and shifts in the ground state absorption spectrum of RB. PMID- 12081316 TI - Formation of singlet oxygen by urocanic acid by UVA irradiation and some consequences thereof. AB - Singlet oxygen-initiated decomposition of urocanic acid (UCA) (3-(1H-imidazol 4(5)-yl)-2-propenoic acid) was used to successfully confirm the report that UCA generates singlet oxygen when irradiated with ultraviolet A light (UVA). The UCA generated singlet oxygen converts UCA to one or more products that then catalyze the further destruction of the UCA with UVA light by singlet oxygen formation. Some nicking of the phiX-174 supercoiled plasmid DNA was observed when UCA was irradiated with UVA to complete destruction of the starting material, and the product mixture was then mixed with the plasmid in the dark. More extensive nicking was seen when the photoproduct mixture and the plasmid were irradiated with UVA light. An "aged" (4 days) solution of UCA photoproduct no longer caused nicking in the dark but retained the capability to nick the plasmid when irradiated. There is evidence for the presence of hydroperoxides in the UCA photolysis product mixture, and the quenching studies with 2-propanol indicate that free radicals are involved in the plasmid-nicking photochemistry. Singlet oxygen does not appear to play a role in the nicking of the plasmid. PMID- 12081317 TI - Heterogeneity of diffusion inside microbial biofilms determined by fluorescence correlation spectroscopy under two-photon excitation. AB - Fluorescence correlation spectroscopy (FCS) under two-photon excitation was applied successfully to characterize the penetration and diffusion capabilities of fluorescent probes (latex beads and fluorescein isothiocyanate-dextran) of different size and electrical charge in two models of monomicrobial biofilms with low (Lactococcus lactis biofilm) or high (Stenotrophonas maltophilia biofilm) contents of extracellular polymeric substance (EPS). FCS measurements performed on each biofilm can show deviation from Brownian diffusion, depending on the local structure of the biofilm and the fluorophore size. In this case, we fitted the data to an anomalous diffusion model and determined apparent diffusion coefficients, which can be 50 times smaller than the values in aqueous solutions. This result was interpreted as steric hindrance of the diffusion of the fluorescent particles within the biofilm that can lead to a total inhibition as observed particularly in the mushroom-like structure of the S. maltophilia biofilm. Alternatively, mechanisms for the absence of FCS signal behavior were related to attractive electrostatic interactions between cationic particles and negatively charged bacteria or to specific interactions between dextrans and EPS of the biofilm matrix. PMID- 12081318 TI - Photoinduced transannular reaction of 1,3-dimethylcyclooctapyrimidine-2,4-diones. AB - This article reports further elaboration of the authors' investigations on the acid-catalyzed photorearrangements undergone by derivatives of 1,3 dimethylcyclooctapyrimidine-2,4-dione. In this case the results are represented for compounds containing a simple methyl or fluorine substituent at different positions in the eight-membered ring. Upon photolysis in frozen benzene containing trifluoroacetic acid, the monomethylated compounds give varying yields of pentalenopyrimidinedione products that are not generated from the fluorinated compounds, which are either inert or undergo hydration accompanied by dehyrofluorination. Plausible mechanisms are advanced to account for the formation of the observed photoproduct species. The work represents an interesting extension and uncovered new aspects of the photorearrangement process. PMID- 12081319 TI - Protective effect of Boldo and tea infusions on the visible light-mediated pro oxidant effects of vitamin B2, riboflavin. AB - The effect of Boldo and black tea infusions on the pro-oxidant effects of vitamin B2, riboflavin (RF), when exposed to the action of visible light was studied. The amounts of antioxidants present in Boldo and tea infusions were evaluated by a procedure based on the bleaching of preformed 2,2'-azinobis(3 ethylbenzothiazoline-6-sulfonic acid) radical cations and were expressed as 6 hydroxy-2,5,7,8-tetramethyl-chroman-2-carboxylic acid equivalent concentrations. The quenching rate constants of singlet oxygen (1O2; [kq]Boldo = 6.0 x 10(7) M( 1) s(-1) and [kq]Tea = 3.2 x 10(7) M(-1) s(-1)) and triplet RF (3RF; [3RFkq]Boldo = 10 x 10(8) M(-1) s(-1) and [3RFkq]TEA = 3.2 x 10(8) M(-1) s(-1)) with Boldo and tea were determined by flash photolysis. These data allow a quantitative interpretation of the results obtained. Our data suggest that most of the oxygen consumption observed in the photolysis of RF in the presence of tea and Boldo infusions is caused by 1O2 reactions. The oxygen consumption quantum yield is considerably smaller than the fraction of RF triplets trapped by the additives (AH) present in the infusion, indicating that their interaction with 3RF does not lead to chemical reactions or that the AH*+ radical ions initially formed participate in secondary processes that do not consume oxygen. Boldo and tea infusions have a significant protective effect when a system containing RF and tryptophan (Trp) is exposed to visible light, not only by quenching the 1O2 and interfering with the Type-I mechanism but also by repairing the damage to Trp molecules associated with the latter mechanism. PMID- 12081320 TI - The quenching of CdSe quantum dots photoluminescence by gold nanoparticles in solution. AB - The photoluminescence (PL) of CdSe quantum dots (QD) in aqueous media has been studied in the presence of gold nanoparticles (NP) with different shapes. The steady state PL intensity of CdSe QD (1.5-2 nm in size) is quenched in the presence of gold NP. Picosecond bleach recovery and nanosecond time-resolved luminescence measurements show a faster bleach recovery and decrease in the lifetime of the emitting states of CdSe QD in the presence of quenchers. Surfactant-capped gold nanorods (NR) with aspect ratio of 3 and surfactant-capped and citrate-capped nanospheres (NS) of 12 nm diameter were used as quenchers in order to study the effect of shape and surface charge on the quenching rates. The Stern-Volmer kinetics model is used to examine the observed quenching behavior as a function of the quencher concentration. It was found that the quenching rate of NR is more than 1000 times stronger than that of NS with the same capping material. We also found that the quenching rate decreases as the length of the NR decreases, although the overlap between the CdSe emission and the NR absorption increases. This suggests that the quenching is a result of electron transfer rather than long-range (Forster-type) energy transfer processes. The quenching was attributed to the transfer of electron with energies below the Fermi level of gold to the trap holes of CdSe QD. The observed large difference between NR and NS quenching efficiencies was attributed to the presence of the [110] facets only in the NR, which have higher surface energy. PMID- 12081321 TI - Modifications of in vitro skin penetration under solar irradiation: evaluation on flow-through diffusion cells. AB - The effect of solar irradiation on ex vivo dermatomed hairless rat skin samples maintained in culture on flow-through diffusion cells for at least 24 h was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and by histological observations. Transepidermal water loss (TEWL) measurements and kinetic analysis of the permeation of both tritiated water and 14C caffeine through the skin were performed after full-spectrum solar exposure involving the use of a xenon arc solar simulator. After a UV exposure of less than 420 mJ/cm2, skin integrity and permeation of both water and caffeine did not change significantly. In contrast, after a 420 mJ/cm2 UV exposure, the epidermis appeared more contracted, associated with an increase of 55% of TEWL and 220% of the skin permeation of tritiated water after 6 h. The data suggested a dramatic alteration of the skin barrier integrity. Moreover, the flux of 14C caffeine increased rapidly by 338% of the absorption of water 12 h after irradiation. These results reveal the presence of a threshold UV exposure that would not modify skin penetration. PMID- 12081323 TI - Photoinhibition in vivo and in vitro involves weakly coupled chlorophyll-protein complexes. AB - In the present study the analysis of the relation between the excited state population in the photosystem II (PSII) antenna and photoinactivation has been extended from an in vitro system, isolated thylakoids, to an in vivo system, Chlamydomonas reinhardtii cells. The results indicate that the excited state quenching by an added singlet quencher induces maximal protection against photoinhibition of about 30% of that expected on the basis of the observed light intensity-treatment time reciprocity rule. Similar results, obtained previously with thylakoids, have been interpreted in terms of damaged or incorrectly assembled complexes that play an important role in photoinhibition in the thylakoid membranes (Santabarbara, S., K. Neverov, F. M. Garlaschi, G. Zucchelli and R. C. Jennings [2001] Involvement of uncoupled antenna chlorophylls in photoinhibition in thylakoids. FEBS Lett. 491, 109-113.). In an attempt to better define this aspect, the photoinhibition action spectra were determined for mutant barley thylakoids, lacking the chlorophyll (Chl) a-b complexes of the outer antenna, and for its wild type. The results indicate that in both systems the action spectra are significantly blueshifted (2-4 nm) and are broader than the PSII absorption in the membranes. These data are interpreted in terms of a heterogeneous population of outer and inner antenna pigment-protein complexes that contain significant levels of uncoupled Chl. PMID- 12081322 TI - Light- and redox-dependent thermal stability of the reaction center of the photosynthetic Bacterium rhodobacter sphaeroides. AB - Irreversible loss of the photochemical activity and damage of the pigments (bacteriochlorophyll [Bchl] monomer, Bchl dimer [P] and bacteriopheophytin) by combined treatment with intense and continuous visible light and elevated temperature have been studied in a deoxygenated solution of reaction center (RC) protein from the nonsulfur purple photosynthetic bacterium Rhodobacter sphaeroides. Both the fraction of RC in the charge-separated redox state (P+Q-, where Q is a quinone electron acceptor) and the degradation of the pigments showed saturation as a function of increasing light intensity up to 400 mW cm(-2) (488/515 nm) or 1100 microE m(-2) s(-1) (white light). The thermal denaturation curves of the RC in the P+Q- redox state demonstrated broadening and 10-20 degrees C shift to lower temperature (after 30-90 min heat treatment) compared with those in the PQ redox state. Similar but less striking behavior was seen for RC of other redox states (P+Q and PQ-) generated either by light or by electrochemical treatment in the dark. These experiments suggest that it is not the intense light per se but the changes in the redox state of the protein that are responsible for the increased sensitivity to photo- and heat damage. The RC with a charge pair (P+Q-) is more vulnerable to elevated temperature than the RC with (P+Q or PQ-) or without (PQ) a single charge. To reveal both the thermodynamic and kinetic aspects of the denaturation, a simple three-state model of coupled reversible thermal and irreversible kinetic transitions is presented. These effects may have relevance to the heat stability of other redox proteins in bioenergetics. PMID- 12081324 TI - Self-assembly of [Et,Et]-bacteriochlorophyll cF on highly oriented pyrolytic graphite revealed by scanning tunneling microscopy. AB - The chlorosomal light-harvesting antennae of green phototrophic bacteria consist of large supramolecular aggregates of bacteriochlorophyll c (BChl c). The supramolecular structure of (3(1)-R/S)-BChl c on highly oriented pyrolytic graphite (HOPG) and molybdenum disulfide (MoS2) has been investigated by scanning tunneling microscopy (STM). On MoS2, we observed single BChl c molecules, dimers or tetramers, depending on the polarity of the solvent. On HOPG, we observed extensive self-assembly of the dimers and tetramers. We propose C=O...H-O...Mg bonding networks for the observed dimer chains, in agreement with former ultraviolet-visible and infrared spectroscopic work. The BChl c moieties in the tetramers are probably linked by four C=O...H-O hydrogen bonds to form a circle and further stabilized by Mg...O-H bondings to underlying BChl c layers. The tetramers form highly ordered, distinct chains and extended two-dimensional networks. We investigated semisynthetic chlorins for comparison by STM but observed that only BChl c self-assembles to well-structured large aggregates on HOPG. The results on the synthetic chlorins support our structure proposition. PMID- 12081325 TI - DNA and protein changes caused by disease in human breast tissues probed by the Kubelka-Munk spectral functional. AB - Malignant, fibroadenoma, normal and adipose breast tissues were studied using diffuse reflectance spectroscopy. The absorption spectra of the breast tissues were extracted from the diffuse reflectance spectra using the Kubelka-Munk function (K-M function). The spectral features of the K-M function were identified and compared with those of the absorption spectra. The spectral features of the K-M function were assigned to DNA, protein, beta-carotene and hemoglobin (oxygenated and deoxygenated) molecules in the breast tissue. The amplitudes of the K-M function averaged from 275 to 285 nm and from 255 to 265 nm and were found to be different for malignant, fibroadenoma and normal tissues. These differences were attributed to changes in proteins and DNA. A set of critical parameters was determined for separating malignant tissues from fibroadenoma and normal tissues. This approach should hold for other tissue types such as cervix, uterus and colon. PMID- 12081326 TI - Near-infrared photosensitizer based on a cycloimide derivative of chlorin p6: 13,15-N-(3'-hydroxypropyl)cycloimide chlorin p6. AB - The 13,15-N-(3'-hydroxypropylcycloimide) chlorin p6 (CIC), which absorbs at 711 nm, possesses considerable photoinduced cell-killing activity. It is 43-, 61- and 110-fold more active than chlorin p6, 3-formyl-3-devinyl chlorin p6 and Photogem, respectively, and has no cytotoxicity without irradiation as estimated on A549 human adenocarcinoma cells. To attain the highest intracellular penetration and activity the monomeric form of CIC should be stabilized. This stabilization in an aqueous environment can be achieved using 0.002-0.005% of Cremophor EL emulsion (polyoxyethylene derivative of hydrogenated castor oil). The intracellular accumulation of CIC occurs in cytoplasm in a monomeric form bound to cellular membranes. This form of the dye is characterized by a high quantum yield of singlet oxygen generation (0.66 +/- 0.02). Besides diffuse staining of intracellular membranous structures, CIC accumulates 3- to 4-fold more intensely in mitochondria and Golgi apparatus, thus indicating these organelles to be the initial targets of its photodynamic action. The incubation time providing 50% accumulation level of CIC in cells is 30 +/- 5 min. The time for 50% release of CIC from the cells is 60 +/- 10 min. A 10-fold decrease in CIC intracellular penetration at 22 degrees C proves that temperature-sensitive mechanisms of transport, rather than diffusion, are responsible for the dye uptake. The average cytoplasmic concentration of CIC was seven times the extracellular concentration in the 0.2-1.6 microM range, used for the photodynamic activity measurements. The concentration of CIC and the light dose that correspond to ca 50% level of phototoxicity induce predominantly an apoptotic-type of cell death, whereas the conditions providing 100% level of phototoxicity induced necrosis. The results obtained indicate that cycloimide derivatives of chlorin p6 may serve as a base for the development of an efficient near-IR photosensitizer. PMID- 12081327 TI - Photodynamic therapy with pyropheophorbide-a methyl ester in human lung carcinoma cancer cell: efficacy, localization and apoptosis. AB - Pyropheophorbide-a methyl ester (MPPa) is a semisynthetic photosensitizer derived from chlorophyll a. The absorption peak of MPPa in organic solvent and in cells was at 667 and 674 nm, respectively. 3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide reduction assay showed that MPPa had no dark cytotoxicity. In vitro photodynamic activity was extensively evaluated using a human lung carcinoma cancer cell line (NCI-h446). MPPa exhibited no genotoxicity, as assayed by single-cell gel electrophoresis. Using confocal laser scanning microscopy and organelle-specific fluorescent probes, MPPa was found to localize in the intracellular membrane system, namely the endoplasmic reticulum, Golgi apparatus, lysosomes and mitochondria, in the NCI-h446 cells. Furthermore, nuclear staining and DNA gel electrophoresis revealed that DNA condensation and fragmentation occurred post-photodynamic therapy, indicating the cell death was in the apoptotic mode. PMID- 12081329 TI - Tuning up or down the UV-induced apoptosis in Chinese hamster ovary cells with cell cycle inhibitors. AB - Exposure to UVC induces apoptosis in Chinese hamster ovary (CHO.K1) cells. While studying the underlying mechanism, we found that a variety of cell cycle inhibitors, including colcemid, hydroxyurea and mimosine, enhance the UV-induced apoptosis in these cells. Such enhancement was not dependent on the cell cycle progression nor was it related to the difference in UV sensitivity at different phases of the cell cycle. The expression of p21(waf1/cip1), a general cyclin dependent kinase (CDK) inhibitor, was deficient in CHO.K1 cells. Ectopic overexpression of the human p21 markedly increased the survival rates of the UV irradiated cells in the presence of colcemid. In addition, roscovitine, a small molecule inhibitor of CDK, also inhibited the UV-induced apoptosis. These observations suggest that deregulation of CDK activity may be critical in the UV induced apoptosis in CHO.K1 cells. PMID- 12081328 TI - The peripheral benzodiazepine receptor in photodynamic therapy with the phthalocyanine photosensitizer Pc 4. AB - The peripheral benzodiazepine receptor (PBR) is an 18 kDa protein of the outer mitochondrial membrane that interacts with the voltage-dependent anion channel and may participate in formation of the permeability transition pore. The physiological role of PBR is reflected in the high-affinity binding of endogenous ligands that are metabolites of both cholesterol and heme. Certain porphyrin precursors of heme can be photosensitizers for photodynamic therapy (PDT), which depends on visible light activation of porphyrin-related macrocycles. Because the apparent binding affinity of a series of porphyrin analogs for PBR paralleled their ability to photoinactivate cells, PBR has been proposed as the molecular target for porphyrin-derived photocytotoxicity. The phthalocyanine (Pc) photosensitizer Pc 4 accumulates in mitochondria and structurally resembles porphyrins. Therefore, we tested the relevance of PBR binding on Pc 4-PDT. Binding affinity was measured by competition with 3H-PK11195, a high-affinity ligand of PBR, for binding to rat kidney mitochondria (RKM) or intact Chinese hamster ovary (CHO) cells. To assess the binding of the Pc directly, we synthesized 14C-labeled Pc 4 and found that whereas Pc 4 was a competitive inhibitor of 3H-PK11195 binding to the PBR, PK11195 did not inhibit the binding of 14C-Pc 4 to RKM. Further, 14C-Pc 4 binding to RKM showed no evidence of saturation up to 10 microM. Finally, when Pc 4-loaded CHO cells were exposed to activating red light, apoptosis was induced; Pc 4-PDT was less effective in causing apoptosis in a companion cell line overexpressing the antiapoptotic protein Bcl-2. For both cell lines, PK11195 inhibited PDT-induced apoptosis; however, the inhibition was transient and did not extend to overall cell death, as determined by clonogenic assay. The results demonstrate (1) the presence of low-affinity binding sites for Pc 4 on PBR; (2) the presence of multiple binding sites for Pc 4 in RKM and CHO cells other than those that influence PK11195 binding; and (3) the ability of high supersaturating levels of PK11195 to transiently inhibit apoptosis initiated by Pc 4-PDT, with less influence on overall cell killing. We conclude that the binding of Pc 4 to PBR is less relevant to the photocytotoxicity of Pc 4-PDT than are other mitochondrial events, such as photodamage to Bcl-2 and that the observed inhibition of Pc 4-PDT induced apoptosis by PK11195 likely occurs through a mechanism independent of PBR. PMID- 12081330 TI - Photoreduction of bacteriorhodopsin Schiff base at low humidity. A study with C13=C14 nonisomerizable artificial pigments. AB - The retinal protonated Schiff base of bacteriorhodopsin is photoreactive to reducing agents such as NaBH4. In the present work we have studied the effect of different protein hydration levels on the photoreductive reaction, as well as the consequences of preventing isomerization around the critical C13=C14 retinal double bond. It was revealed that the rate of light-induced NaBH4 reaction can be fitted to three phases, between 100 and 87%, from 87 to 35% and below 35% relative humidities (r.h.). The three phases are attributed to three protein regions characterized by different water affinities. Furthermore, it is shown that the PSB reduction reaction is light catalyzed even in artificial pigments derived from retinal analogs, in which isomerization around the C13=C14 double bond is prevented. It is suggested that the protein experiences light-induced conformational alterations that are not associated with C13=C14 double bond isomerization. In the 13-cis locked pigment the rate of reduction reaction is affected by r.h. levels only below 35%. The relatively low r.h. required for withdrawing water from the protein is attributed to the increased protein-water affinity in this specific pigment. PMID- 12081331 TI - Regulation of cAMP-mediated photosignaling by a phytochrome in the cyanobacterium Anabaena cylindrica. AB - Changes in cellular adenosine 3',5'-cyclic monophosphate (cAMP) content induced by monochromatic light of various wavelengths were determined in the cyanobacterium Anabaena cylindrica. Irradiation with monochromatic red light caused a rapid decrease in cAMP content. In contrast, far-red light caused a rapid increase in its content. The effects of red and far-red light were reversible, suggesting the involvement of a prototype phytochrome as the photoreceptor for cAMP-mediated light-responsive signal transduction. PMID- 12081332 TI - The damage repair role of He-Ne laser on plants exposed to different intensities of ultraviolet-B radiation. AB - Light-grown broad bean (Vicia faba L.) seedlings were subjected to different intensities of UV-B radiation (0, 0.05, 0.15, 0.45, 0.90, 1.45 and 1.98 W m(-2)) for 7 h under photosynthetically active radiation (70 micromol m(-2) s(-1)) and then exposed to He-Ne laser (632.8 nm, 5.43 mW mm(-2)) radiation for 5 min or red light radiation for 4 h without ambient light radiation. When He-Ne laser radiated leaves were treated using lower intensity UV-B, the activities of superoxide dismutase (EC 1.15.1.1), ascorbate peroxidase (EC 1.11.1.11) and catalase (EC 1.11.1.6) improved significantly. Moreover, the UV-B-injured plants treated with laser light recovered faster from UV-B treatment because the concentration of malondialdehyde and the rate of electrolyte leakage from leaf disks reached control levels (no UV-B or laser treatment) early compared with those exposed only to ambient light or in dark conditions. Laser treatment, however, had no repair effect on seedling damage induced by higher UV-B radiation (1.45 and 1.98 W m(-2)), even with higher laser flux rates and longer laser treatment. In addition, the red light treatment had no repair effect on UV-B induced damage. Meanwhile, the long-term physiological effect of He-Ne laser treatment on UV-B damaged plants was presented and evaluated. The results showed that the laser had a long-term positive physiological effect on the growth of UV B-damaged plants. With the exception of the severe damage caused by higher UV-B radiation, a laser with the proper flux rate and treatment time can repair UV-B induced damage and shorten the recovery time. PMID- 12081333 TI - Endoscopic evaluation of upper abdominal symptoms in adult patients, Saudi Aramco Ai Hasa Health Center, Saudi Arabia. AB - Upper oesophago-gastro-duodenoscopies (UGI endoscopies) were performed on adult patients who attended Saudi Aramco Health Centre at Al Hasa, Saudi Arabia, between June 1986 and December 1993, with complaint of upper abdominal pain. During this period, three hundred and fifteen (315) patients were examined. Sixty one percent were females and the rest males. Peptic ulcer disease including gastritis was the most common diagnosis made. Helicobacter pylori-like organisms were documented historically in biopsy samples in 113 of 178 (63%) patients. Nearly all were associated with gastritis; no organisms were reported in normal histology specimens. This confirms the association of this organism with gastritis. Gastric cancer suspected radiologically were not confirmed endoscopically or histologically. It is important to endoscope and biopsy for histology suspected gastric cancer and ulcer; a negative result may save the patient from unnecessary surgery, ideally all normal looking gastric mucosa should be biopsied so that histological gastritis is not missed. PMID- 12081334 TI - Aetiological significance of alcohol in liver cirrhosis on the Jos Plateau. AB - Fifty-one patients with histologically diagnosed cirrhosis were studied prospectively over one year period. There were forty-one males and ten females. The majority of patients (75.59%) took alcohol in significant quantity and it was thought to be the most likely cause of the cirrhosis in them. Eighty percent of them had micronodular cirrhosis and 51.3% of them drank only "burukutu" a local alcoholic brew brewed mostly in iron pots and contains two other hepatotoxic agents--aflatoxin and iron. Eight (5.7%) of the patients and five (9.8%) of controls were positive for hepatitis B surface antigen (HBs.Ag). It is likely that alcohol is very important cause of cirrhosis of the liver seen in Jos. PMID- 12081335 TI - A prospective evaluation of Medizip in Nigeria. AB - In a one year period between April, 1997 and March 1998 we have carried out a randomized controlled prospective study to evaluate the outcome of Medizip Surgical Zipper technique in closing 50 surgical wounds in Nigeria. There were similar sex, age and wound distribution between the Medizip study group and the conventional (needle and thread) control group. The cosmetic outcome of scar was rated as good or bad. It was good if the emerging scar was a thin linear (< or = 2 mm wide) scar, and bad if the scar was a broad flat (> 2 mm wide) or a heap-up (hypertrophic or keloidal) scar. The cosmetic outcome of scar was good in 45 (90%) of the Medizip study group as compared with 22 (44%) of the conventional control group. The observed difference between the two groups was statistically significant (P < 0.05). We observed superficial wound infection in one (2%) of the Medizip study group as compared with 10 (20%) of the conventional control group. The results of our study clearly demonstrated that the use of Medizip surgical Zipper would be a superior technique of closing surgical wounds in terms of the cosmetic outcome of scars and associated problems. PMID- 12081336 TI - Hospital prevalence of pulmonary tuberculosis and co-infection with human immunodeficiency virus in Ilorin: a review of nine years (1991-1999). AB - This is a hospital based retrospective study that determines the prevalence of pulmonary tuberculosis (PTB) and seroprevalence of Human Immunodeficiency Virus (HIV) infection amongst PTB cases. The study was over a nine year period. The annual distribution of all PTB and PTB/HIV co-infected cases were estimated, so was the annual distribution of all medical cases seen over the same period. A total of 104,240 medical cases were seen, out of which 9697 were confirmed PTB. This gives a prevalence rate of 9.2%. Their sex ratio was M:F 1.3:1 and about 81.6% of them were between 15-54 years of age. Only 189 out of 1496 cases of PTB screened for HIV antibody were seropositive giving a seroprevalence rate of 12.6%. The annual distribution of PTB/HIV co-infection showed a rising trend from only one case in 1991 to 20 cases in 1999. Majority of those who were seropositive were traders, housewives and drivers. PMID- 12081337 TI - The effect of parental presence at induction of anaesthesia on the behaviour of unsedated Nigerian children presenting for day-case anaesthesia. AB - The effect of parental presence at induction of anaesthesia on one hundred and eighteen children presenting for day-case inguinal operations and the behaviour of the attending parents at induction were studied. Of the 24 children aged 5 years and below whose parents at were present at induction, 25% were distressed, while 80% of 36 children of this age group whose parents were absent showed distress. Better behaviour was observed among the children who were above 5 years of age. When parents were present 7% of 28 children cried while 20% of 30 children cried when parents were absent. In both age groups induction heart rates were significantly higher in the children whose parents were absent. All parents except one who cried and another one who was too anxious, were helpful at induction and they would welcome the same opportunity to be present at the children's induction. PMID- 12081338 TI - Spectrum of paediatric malignancies in eastern Nigeria (1989-1998). AB - A 10-year retrospective survey of childhood malignancies in eastern Nigeria was carried out based on data from the University of Nigeria Teaching Hospital Cancer Registry. The aim was to discover the current pattern of paediatric malignancies in the study area. A total of 313 children aged less than 15 years, diagnosed with cancer by means of histological or cytological examination during the study period (January 1989-December 1998) were analyzed. The male-to-female ratio was 1.6:1. The lymphomas constituted the highest prevalence (38.3%) with Burkitt's lymphoma being the commonest (65.8%) of all the lymphomas. These were followed by sarcomas (14.7%), nephroblastoma (14.4%), retinoblastoma (12.1%) and leukaemias (8.6%). The less common ones were teratomas (2.9%) and neuroblastoma (1.9%). Comparison of two clinicopathological studies of childhood cancer in Enugu between 1976-1980 and 1989-1998 showed an increase in the relative frequencies of sarcomas, retinoblastoma, leukaemias and a decline in the frequencies of Burkitt's lymphoma, nephroblastoma, teratomas and neuroblastoma. Further epidemiological studies are necessary in order to determine whether these changes actually reflect the distribution of childhood cancer in the general population and also to elicit the possible mechanisms of carcinogenesis. PMID- 12081339 TI - Treatment outcome of otomycosis in Ilorin, Nigeria. AB - Among 141 patients suspected of having otomycosis, 76 (53.9%) were mycologically confirmed. The fungi isolated were Aspergillus sp (63.4%), Candida (35.5%) and Mucor (1.3%). Ninety-six per cent were symptom free within 2 weeks of topical application of 1% clotrimazole cream, after thorough failures in the cleaning of debri in the ear canal. Treatment failures were minimal, including recurrence (2.6%), acute otitis externa (1.3%), foreign body in the ear (1.3%) and blocking of ear by therapeutic agent (2.6%). This treatment regimen is simple, efficacious, cost effective and safe; hence it is recommended for adoption in the management of otomycosis. PMID- 12081340 TI - Assessment of educational services available to blind and low vision school children in Lagos, Nigeria. AB - A descriptive study to assess the educational services available to children attending the Pacelli School for the Blind in Lagos, Nigeria. A standardised methodology was used with structured and semi-structured questionnaires. Observations were also made in some areas. Of the 26 children aged 10-15 years who were identified as having low vision, 15 (57.7%) of them had significant improvement in distance and near vision after refraction while one third (31%) had improvement in their reading with low power magnifying lenses. All except 4 (25%) teachers were well educated but none seemed to have appreciable knowledge about the teaching of low vision children as all these children were taught Braille like their blind peers. While educational facilities available were reasonably adequate for blind children, no visual aids were available for low vision children. It is recommended that appropriate educational facilities, optical and non-optical devices should also be provided particularly for low vision children to enable them achieve their optimal potential in life. PMID- 12081341 TI - Internal fixation at ECWA Hospital, Egbe, Kogi State, Nigeria. AB - Forty two (42) cases of fractures treated by internal fixation at Egbe Hospital ECWA, Kogi State, Nigeria were analysed retrospectively. Most of the patients requiring internal fixation were young male patients in their reproductive years 26 patients were under 50 years of age out of which 22 patients were males (84.6%) whilst only 4 patients (15.4%) were females. Internal fixation was found to be commoner in males: 31 patients (73.8%) than in females: 11 patients (26.2%). Fractures requiring internal fixation are commoner in the lower limbs than in the upper limbs (Ratio 6:1). The femur is the commonest site of internal fixation, 22 cases (54.2%) followed by the tibia, 6 cases (14.3%). Majority of the patients, 28 cases (67%) left the hospital under 50 days whereas when treated conservatively most will be in the hospital for up to 100 days. Only about 60% of the cases require blood transfusion and even a more cautious approach could reduce this percentage. 13 (31%) of the cases had fever post-operatively which was not necessarily due to infection. Based on its advantages, internal fixation is therefore encouraged and should only be performed by a surgeon who has adequate experience, equipment, instruments and a good operating theatre setting. PMID- 12081342 TI - Beliefs and perceptions about HIV infection and AIDS among mothers of infants in Ibadan, Nigeria. AB - In view of the growing importance of the human immunodeficiency virus (HIV) infection and the Acquired Immunodeficiency Syndrome (AIDS) in Nigeria, 221 mothers were interviewed by means of semi-structured questionnaires in an infant welfare clinic in Ibadan, Nigeria, about their perceptions and beliefs about HIV/AIDS infection with particular emphasis on how the infection could be acquired, how the risk of its acquisition can be reduced and whether how children could be infected. The findings revealed that 81% of the respondents believe that HIV infection does exist in Nigeria but only 17.6% think they have seen an AIDS patient. About two-thirds (68.8%) believe that children can be infected with HIV, 70.6% believe that a mother can transmit the infection to her child and 58% believe that HIV can be transmitted through breast milk. Eighty percent believe that people can protect themselves from having the infection by various means, including avoiding sharing needles or reusing hypodermic needles (83.3%), receiving unscreened blood (79.2%), mutual fidelity with a single sexual partner (73.8%), use of condoms (67.9%), avoiding the use of public toilets (56.1%) and avoiding mosquito bites (35.3%). Those with higher education showed better knowledge than those with lower education. In spite of the strong correlation with formal education, some misconceptions existed in all groups. For example, those without any formal education compared with those with some formal education believed that use of public toilet can cause HIV to be transmitted, the difference was not statistically significant (chi2 = 8.87; p = 0.064). Interestingly more of the highly literate women believed that HIV infection can be acquired by mosquito bites (chi2 = 16.82; p = 0.002). It was concluded that awareness of HIV infection and AIDS is high among mothers of infants attending the infant welfare clinic of the Institute of child Health, Ibadan. Whatever educational interventions is planned for this facility should take these findings into account and specially target those with none or little formal education. PMID- 12081343 TI - Carriage of antimicrobial resistant Escherichia coli in adult intestinal flora. AB - Knowledge of antibiotic resistance in bacteria strains colonizing healthy people is important for several reasons, one of which is that; these organisms form one of the largest reservoirs of resistant genes. Frequency of resistance to eleven different antimicrobial agents was examined in faecal flora of adults with no history of recent antimicrobial treatment. Using the disc diffusion sensitivity test, 106 strains of Escherichia coli were examined, 68% of these were resistant to tetracycline, and 57% were resistant to ampicillin and cotrimoxazole respectively. There was no resistance to cefuroxime but resistance to ceftazidime was 13%. Fifty six out of the eighty eight (64%) isolates, which showed any resistance, were resistant to three or more antimicrobials. The most common resistant pattern was to three drugs tetracycline, ampicillin and cotrimoxazole. Six strains were susceptible to all antibiotics. One strain of Escherichia coli was resistant to eight antimicrobials. Thirty per cent of the Escherichia coli were resistant to gentamicin. This study reveals a high prevalence of resistant bacteria in faecal flora of healthy adults. PMID- 12081344 TI - Residual and recurrent parotid gland neoplasm after surgical excision. AB - Ten patients who presented with residual or recurrent parotid gland neoplasm after elective curative surgical excision between January 1992 to December 1999 were studied. It aimed at finding the contributory factors responsible for the persistence or recurrence of the tumour after surgical excision. The 10 patients comprised of four males and six females. Their ages ranged from 33 to 73 years with a mean of 52 years. Seven were of malignant histologic cell types while three were benign. Eight patients were treated with secondary parotidectomy while two received combination chemotherapy. In five patients, both the superficial and deep lobes of the parotid gland were found during secondary parotidectomy. This confirms that the extent of the initial surgery was less than superficial parotidectomy. This confirms that the extent of the initial surgery was less than superficial parotidectomy. Four patients were later referred to radiotherapy centers for further treatment It is concluded that most recurrent parotid neoplasm is a result of inadequate primary surgical excision, and best results are obtained by doing a minimum of superficial parotidectomy in the surgical treatment of parotid gland neoplasms. PMID- 12081345 TI - An epidemiological survey of the use of cosmetic skin lightening cosmetics among traders in Lagos, Nigeria. AB - The use of skin lightening creams has become a socially acceptable phenomenon widely practised by both men and women in Lagos, Nigeria. Four hundred and fifty traders were selected to ascertain the extent of this practice and what socio cultural and economic factors determine the use of these cosmetics. Prevalence of use of skin lightening cosmetics was 77.3% comprising ninety-six males (27.6%) and two hundred and fifty two females (72.4%). The trend cuts across all socioeconomic strata, age, sex and marital status. Hydroquinolone based products were the most commonly used products although cortico-steroids and mercury-based products were also widely used. Of eleven dermatological side effects that were reported, exogenous ochronosis was the commonest. Recommendations on how to correct this ill in the society have been proffered. PMID- 12081346 TI - Should routine ultrasonography in pregnancy be mandatory? AB - A review of the first 500 obstetric ultrasound scans performed at the National Hospital, Abuja between September 1999 and February 2000 was carried out. Five hundred pregnant women aged 18-41 years, mean age of 30 +/- 3.41 years; who had obstetric scans were studied. 287 (57.4%) had routine scans while 213 (42.6%) had specific obstetric reasons for scanning. The former served as the control group. Of the total, positive findings were seen in 180 cases (36%). 71.8% of those with specific indications for scanning had positive findings of clinical significance, while only 9.4% of the control group had incidental findings of clinical significance. Positive findings on ultrasound were significantly greater in the group with specific reasons for scanning, P < 0.001. We suggest that ultrasound examination in pregnant women should be performed based on established medical indications, elucidated clinically. PMID- 12081347 TI - How to adapt an ordinary drilling machine for orthopaedic surgery. AB - An orthopaedic drilling machine has number of revolutions per minute (r/min) < or = 750. This low r/min. accounts for low friction and less tissue damage during bone drilling. This is an advantage over the ordinary drilling machine whose r/min. > 2800. A method of converting it into an orthopaedic drilling machine is described here. PMID- 12081348 TI - Spontaneous intracranial haemorrhage: computed tomographic patterns in Accra. AB - The diagnosis of stroke and the ability to distinguish the subtypes is central in the management of patients. This CT study has confirmed an increased prevalence of stroke (CVA) among Ghanaians. It has also reaffirmed a relatively higher incidence (52.9%) of spontaneous intracranial haemorrhage over cerebral infarcts among the 1,172 stroke patients studied. The study points to a male preponderance and a mean age of 55.7 years. Parenchymal haemorrhage was found to be the commonest variety of these haemorrhages. It occurred in 83.6% of cases while primary subarachnoid haemorrhage was reported in only 8.1% of cases. Ventricular extension of the parenchymal haemorrhage was reported in 22.7% of cases. The latter were mostly unilateral and on the left side especially in the parietal lobe (70.9%), subdural haemorrhage like the parenchymal variety was also reported to be more on the left, mainly unilateral and acute. Haemorrhages in the cerebellum and pons which are normally difficult to diagnose were also outlined with ease in the CT images. Other CT findings in these patients include parilesional oedema and mass effect found in 87.10% and 77.4% respectively. PMID- 12081349 TI - Causes of low vision and blindness in children in a blind school in Lagos, Nigeria. AB - A descriptive study to determine the causes of low vision and blindness in children attending the Pacelli School for the Blind in Lagos State, Nigeria. A standardised methodology was used with structured and semi-structured questioinaires. Twenty-six children aged below 16 years who were identified as having low vision and blindness were examined to determine the causes of the low vision and blindness. The anatomical sites of diseases leading to low vision and blindness in these children were retina (30.8%), lens (23.1%), glaucoma (19.2%), cornea (11.5%) and optic nerve (7.7%). Aetiologically, 38.5% of low vision was due to hereditary factors, 23.1% intra-uterine, 15.4% others and unknown in 23%. Retinal dystrophy was the most common cause of low vision and blindness, while congenital cataract and glaucoma were the major causes of avoidable blindness. Very few cases of corneal scar (Measles/Vitamin A deficiency) were seen. The causes of low vision appeared to be different from those of blindness as hereditary and intra-uterine factors were mainly responsible for low vision. It is recommended that these children be identified early, through low vision care programmes and those with avoidable causes treated accordingly. PMID- 12081350 TI - Exclusive breast-feeding practice and associated factors in Enugu, Nigeria. AB - A cross-sectional questionnaire survey was conducted among 235 infant-mother pairs in five Baby Friendly pairs in five Baby Friendly Hospitals in Enugu Nigeria in 1998. The aims were to study their breast-feeding practices and associated factors. The exclusive breast-feeding rate was 33.3% while the predominant breast-feeding rate was 50.2%. Factors associated with exclusive breast-feeding included infants' birth order (P = 0.015), fathers' education (P =0.0244), mothers' education (P = 0.000001), occupation (P = 0.0069) and parity (P = 0.004). However, the infants' age (P = 0.054) and sex (P = 0.403), mothers' age (P = 0.2005), number of breast-feeding counseling attendances (P = 0.0883) and the breast-feeding initiator (P = 0.473) were comparable irrespective of breast-feeding practice. In the mothers' perspectives, the commonest reasons for not breastfeeding exclusively included; insufficient breast milk (58,37.0%) and the sociocultural practice of giving water to babies because of the hot climate (52,33.1%). For an improvement in the exclusive breast-feeding rate of this population, health workers should highlight to mothers the dangers of water supplementation and the dynamics of breastmilk supply through health education, home visits and the formation of community based lactation support groups. PMID- 12081351 TI - Injection safety: knowledge and practice among health workers. AB - The increase in the spread of human immune-deficiency virus (HIV) infection among our population calls for measures to check its transmission. Health workers are at risk especially those in clinical practice who work with needles and other sharp objects that could aid transmission of the virus. Professionalism is at its lowest at Primary Health Care level and workers responsible for giving immunization with injections need clear understanding of principles of safe injection. To appreciate how to formulate a training programme, the knowledge and practice of injection safety among the workers were assessed through the use of a questionnaire. Fifty eight percent thought only children receiving immunization were at risk of a disease, 58.3% know the diseases transmissible by needle stick injury. On reasons for prevalence of unsafe injection practices, 27% suggested inadequate provision, 18.3% poverty, 17.7% ignorance, 17.2% improper disposal, 11.8% lack of surveillance and health supervision for workers and 8% negligence. On syringes and needles disposal, 53.2% suggested burning only, 2.1% burying and 44.7% both burning and burying. Less than 9% knew that auto-destruct syringes exist while 38.9% were aware of the joint WHO and UNICEF statement on "bundling approach" in vaccine packaging. Only about 50% of respondents had facilities for disposal of needles and syringes in their health centers, 45.6% wear protective clothing during service delivery. On making injection safer 33.7% suggested re training of health workers, 22% retraining of workers and public re-education, 16.8% suggested planning budgeting and purchasing of auto-destruct syringes, 16.5% social mobilization and advocacy and 11% suggested the provision of facilities for disposal. It was concluded that awareness on safe injection practices was low among the health workers and that their practices could put them at greater risk of needles stick injury and hence the spread of HIV and other infection among them. Training on safe injection practices is therefore recommended. PMID- 12081352 TI - A ten year review of maternal mortality in Sokoto, northern Nigeria. AB - OBJECTIVE: To determine the institutional maternal mortality ratio, risk factors, causes and to suggest ways of preventing or reducing the maternal deaths. SUBJECTS & METHODS: A descriptive study based on hospital case records of all maternal deaths at the Usmanu Danfodiyo University Teaching Hospital, Sokoto, between January 1990 and December 1999. RESULTS: There were 197 maternal deaths and 9,158 live births during the study period. The maternal mortality ratio was 2,151/100,000 live births. Mean age at death was 27 years. Risk factors included nulliparity, poverty; illiteracy and lack of prenatal care. Main causes of death were ruptured uterus, eclampsia, infection and haemorrhage. CONCLUSION: The maternal mortality ratio is high. Most of the deaths could have been prevented by prenatal care and prompt treatment of the obstetric emergencies. Unless somebody, somewhere, is prepared to cater for the poor at times of dire need, the maternal death toll will remain high. PMID- 12081353 TI - Inflammatory orbital pseudotumor simulating retinoblastoma in a one year old girl. AB - The case report of a one year old girl with a clinical diagnosis of retinoblastoma who following enucleation was histologically diagnosed as a case of inflammatory orbital pseudotumour is presented. The literature is reviewed and the conclusion is reached that inflammatory orbital pseudotumour be considered in the list of deferential diagnosis of proptosis in early childhood. PMID- 12081354 TI - Matchet cut injury at the wrist as a cause of traumatic radial artery aneurysm: a case report. AB - A sixty year old Yoruba farmer presented at Ela Memorial Medical Centre, Ilorin with recurrent spontaneous bleeding from traumatic left radial artery aneurysm. This was due to a clash between a Fulani Cattle rearer and the Yoruba farmer. This sort of situation is rampant across Nigeria. It has led to unnecessary death and bodily damage in several cases. The sixty year old man was cut on the left wrist with a matchet. This was poorly treated and this resulted into a saccular radial aneurysm which was complicated with recurrent bleeding. Careless treatment of vascular damage could result into aneurysm formation. Medical personnel treating injuries should be aware of vascular injuries and refer the patient promptly for appropriate management. Communal disputes should be settled quickly before they lead to communital clashes which could lead to vascular injuries for which surgeons who could cope with such injuries are far-in-between in Nigeria. PMID- 12081355 TI - The vermiform appendix in the inguinal canal. An unusual abnormal position--a case report and review of literature. AB - A three year old boy was operated on for a right communicating hydrocele. The vermiform Appendix was found in the inguinal canal, outside the hydrocele sac. Appendicectomy was performed. This is an abnormal position for an appendix. PMID- 12081356 TI - HIV-related oral diseases in Benin City, Nigeria. AB - A prospective study, carried out at the University Teaching Hospital Benin City, in which one hundred and twenty-three patients with confirmed HIV/AIDS were examined showed that sixty patients (48.8%) had oral diseases; fungal (80.0%), bacterial (28.3%), viral (10.0%), neoplastic (11.6%), recurrent oral ulcers (10.0%) and non-recurrent oral ulcers (3.3%). The common sites of involvement are also described. The oral manifestations of HIV/AIDS among Nigerians were similar to those reported from other regions. The need for thorough oral examination as part of the general physical examination of patients is emphasized. PMID- 12081357 TI - Using marketing research methods to evaluate a stage-specific intervention. AB - OBJECTIVE: To show how marketing methods can be used to distribute and evaluate a health promotion intervention. METHODS: Mass media promotion was used to communicate a physical activity resource. Brief telephone interviews were used to screen callers and recruit participants into a controlled trial. Follow-up was conducted 3 months later. RESULTS: Information was gained about the attitudes and motivation of callers. The majority of participants (study and control) made significant changes in their activity levels. CONCLUSION: The study demonstrated that even when mass media channels are used, market segmentation can be achieved and program evaluation conducted. PMID- 12081358 TI - Developing effective school-based drug abuse prevention programs. AB - OBJECTIVE: To research effective drug prevention strategies for school-aged populations from drug prevention programs funded by the USDHHS Center for Substance Abuse Prevention (CSAP). METHOD: Nine directors and staff members from model CSAP-funded programs were interviewed. RESULTS: Fifteen strategies that focus on building trusted relationships, selecting well-qualified staff using existing communication networks, and providing timely evaluation feedback are discussed, with examples from the agencies cited. CONCLUSION: Formulating effective partnerships that support both the goals of the outside agency with drug prevention resources and the mission of a school can contribute toward effective school-based drug prevention programs. PMID- 12081359 TI - Rural elementary students', parents', and teachers' perceptions of bullying. AB - OBJECTIVE: To examine the prevalence and correlates of bullying in 7 rural elementary schools from students', parents', and teachers' perspectives. METHOD: Surveys were completed by 739 fourth, fifth, and sixth grade students, 367 parents, and 37 teachers. RESULTS: Students tended to report higher prevalence of bullying than did parents or teachers, and their reports were associated with aggression, attitudes toward violence, and perceptions of school safety. CONCLUSION: Bullying behavior is prevalent in rural elementary schools and is indicative of aggression and proviolence attitudes. Parents and teachers need to pay closer attention to bullying behavior among schoolchildren and to impart their knowledge to children in a comprehensive, coordinated manner. PMID- 12081360 TI - Seat-belt use by trauma center employees before and after a safety campaign. AB - OBJECTIVES: To determine whether employees of a regional trauma center wore seat belts any more often than did visitors to the medical center and residents of the state. To demonstrate whether an intensive safety campaign would improve seat belt compliance among trauma center employees. To determine the duration of improvement. METHODS: Hospital employees and visitors were observed as they exited the medical center's parking ramps over a 3-month period. RESULTS: After a hospital-wide seat-belt campaign, employee compliance rose by 7.5%, to 81.5% at 14 days, but fell back to preintervention levels at one month (76.7%) and 3 months (77%) after the intervention. CONCLUSIONS: An intensive seat-belt safety campaign resulted in only modest and transient improvement in the rate of seat belt use. PMID- 12081361 TI - Survey development for assessing correlates of young adolescents' eating. AB - OBJECTIVE: To describe the development and psychometric testing of a survey to assess multiple influences on young adolescents' eating behaviors. METHODS: A 204 item survey was piloted with middle-school students. We incorporated a broad view of determinants of eating behavior in an effort to expand the current field of predictors. RESULTS: The survey was reduced to 147 items and administered to 3,878 students. Internal consistencies were generally >0.70. Test-retest reliabilities were 0.30-0.98, mostly >0.60. CONCLUSIONS: We estimated psychometric properties for a range of measures in a diverse sample. This is important as the increasing demand for outcomes-based research expands the need for psychometrically sound survey measures. PMID- 12081362 TI - Clarifying the contribution of subjective norm to predicting leisure-time exercise. AB - OBJECTIVE: To clarify the contribution of subjective norm to exercise intention and behavior by considering the influence of descriptive as well as injunctive social norms related to family and friends. METHODS: A sample of 530 college students completed a questionnaire that assessed descriptive and injunctive social norms related to family and to friends, perceived behavioral control, attitude, intention, and leisure-time exercise. RESULTS: Friend descriptive social norm was a significant predictor of both intention (p<.05) and leisure time exercise (p<.001). CONCLUSION: Descriptive norms should be incorporated into tests of the theory of planned behavior in the exercise domain. PMID- 12081363 TI - Oral sexual experience among young adolescents receiving general health examinations. AB - OBJECTIVE: To examine whether oral sex occurs among young adolescents receiving general health examinations and warrants inclusion in sexual risk assessments. METHODS: Confidential, interviewer-assisted questionnaires completed by 12 to 15 year-olds (n=335). RESULTS: Overall, 18% of adolescents reported having oral sex. Among adolescents who had oral sex, 25% reported no vaginal sex. Few adolescents reported using barrier protection during oral sex. Most adolescents thought that penile-vaginal sex (96%) and penile-anal sex (81%) can transmit HIV, but 68% thought that penile-mouth sex can transmit HIV. CONCLUSION: Unprotected oral sex among young adolescents warrants attention in research and clinical sexual assessment. PMID- 12081364 TI - An Arabidopsis callose synthase. AB - Beta-1,3-glucan polymers are major structural components of fungal cell walls, while cellulosic beta-1,4-glucan is the predominant polysaccharide in plant cell walls. Plant beta-1,3-glucan, called callose, is produced in pollen and in response to pathogen attack and wounding, but it has been unclear whether callose synthases can also produce cellulose and whether plant cellulose synthases may also produce beta-1,3-glucans. We describe here an Arabidopsis gene, AtGsl5, encoding a plasma membrane-localized protein homologous to yeast beta-1,3-glucan synthase whose expression partially complements a yeast beta-1,3-glucan synthase mutant. AtGsl5 is developmentally expressed at highest levels in flowers, consistent with flowers having high beta-1,3-glucan synthase activities for deposition of callose in pollen. A role for AtGsl5 in callose synthesis is also indicated by AtGsl5 expression in the Arabidopsis mpk4 mutant which exhibits systemic acquired resistance (SAR), elevated beta-1,3-glucan synthase activity, and increased callose levels. In addition, AtGsl5 is a likely target of salicylic acid (SA)-dependent SAR, since AtGsl5 mRNA accumulation is induced by SA in wild type plants, while expression of the nahG salicylate hydroxylase reduces AtGsl5 mRNA levels in the mpk4 mutant. These results indicate that AtGsl5 is likely involved in callose synthesis in flowering tissues and in the mpk4 mutant. PMID- 12081365 TI - Apple has two orthologues of FLORICAULA/LEAFY involved in flowering. AB - Two orthologues of FLORICAULA/LEAFY AFL1 and AFL2 (apple FLO/LFY), were isolated from the floral buds of apple trees. Their expression was detected in various tissues and during differentiation of the floral buds. Furthermore, the flowering effectiveness of each gene was assessed with transgenic Arabidopsis. Both AFL1 and AFL2 showed high homology to each other (90%) and a high degree of similarity to PTLF and PEAFLO (70%), which are homologues of FLO/LFY from poplar and pea, respectively. RNA blot analysis showed that AFL1 was expressed only in the floral bud during the transition from vegetative to reproductive growth, whereas AFL2 was expressed in vegetative shoot apex, floral buds, floral organs and root. Genomic Southern analysis showed that apple had other homologues in addition to AFL1 and AFL2. The transgenic Arabidopsis with over-expressed AFL2 showed accelerated flowering and gave rise to several solitary flowers from rosette axils directly. AFL1 had similar effects, but the phenotypes of the transgenic Arabidopsis with AFL1 were weaker than those with AFL2. These results suggest that both genes are involved in flower differentiation in apple. PMID- 12081366 TI - Characterization of cis-regulatory regions responsible for developmental regulation of the gibberellin biosynthetic gene GA1 in Arabidopsis thaliana. AB - The Arabidopsis GA1 gene encodes copalyl diphosphate synthase, which catalyzes the first committed step in the gibberellin biosynthetic pathway. Previous studies indicated that the expression pattern of the GA1 gene is tissue-specific and cell-type-specific during development. Here we showed that expression of GA1 cDNA driven by the 2.4 kb 5'-upstream sequence plus the GA1 genomic coding region into the third exon was able to rescue the gal-3 mutant phenotype. To understand the mechanism controlling GA1 gene expression, cis-regulatory regions in the GA1 promoter were identified by promoter deletion analysis with the GA1-beta glucuronidase (GUS) gene fusion system. The second intron and the region from 1391 to -997, with respect to the translation initiation site, positively regulate overall GA1-GUS expression level in all tissues examined. Several additional regulatory regions are involved in GA1-GUS expression in all the stages except in seeds: two positive regulatory regions in the first intron and the sequence between -425 and -207, and a negative regulatory region between 1848 and -1391. We also found that the region between -997 and -796 is essential for a high level of GA1 expression in developing seeds. PMID- 12081367 TI - Ntlim1, a PAL-box binding factor, controls promoter activity of the horseradish wound-inducible peroxidase gene. AB - To understand molecular mechanisms underlying wound-induced expression of plant peroxidase genes, the promoter of a horseradish C2 peroxidase (prxC2) gene was analyzed. We had previously isolated a tobacco nuclear protein, Ntlim1, as a trans factor binding to a PAL-box motif of the prxC2 promoter; however, the function of the Ntlim1 trans factor and the PAL-box motif in wound-responsive expression of the prxC2 gene remains unclear. Here, we found that the prxC2 promoter without the intact PAL-box motif failed to direct a normal level of both the basal and the wound-induced expression of beta-glucuronidase (GUS) reporter gene in transgenic tobacco plants, indicating that the PAL-box motif functions as an essential cis element of the prxC2 promoter. We also found that antisense expression of Ntlim1 in transgenic plants carrying the prxC2 promoter::GUS chimeric construct decreased not only the level of the basal and the wound induced expression of the GUS reporter gene but also the extent of wound inducibility of the prxC2 promoter itself. This result indicates that Ntlim1 is required for the basal level of prxC2 promoter activity as well as its up regulation under wound stress. Moreover, consistent with the results obtained in planta, result from super-shift assay indicates that the Ntlim1 binds to the PAL box motif independently of wound stress. PMID- 12081368 TI - Characterization of five novel dehydration-responsive homeodomain leucine zipper genes from the resurrection plant Craterostigma plantagineum. AB - Homeodomain leucine zipper (HDZip) genes encode putative transcription factors that are unique to plants. A function in regulating processes that are specific for plants is postulated, such as responses to environmental cues and developmental signals. This is supported by a growing body of evidence resulting from studies of HDZip genes in a variety of species. In addition to the previously isolated CPHB-1 and -2 genes, this paper reports the isolation of members of five families of Craterostigma plantagineum homeobox leucine zipper genes (CPHB) via a yeast one-hybrid screening approach. Based on the sequence homology and protein interactions the encoded proteins (CPHB-3/4/5/6/7) were classified into HDZip class II and I genes. Homo- and heterodimerization of CPHB proteins within the same structurally related class has been demonstrated and the DNA-binding activity of CPHB proteins to two homeodomain recognition elements (HDE1 and HDE2) has been compared in yeast. All families of CPHB genes were modulated in their expression in response to dehydration in leaves and roots. CPHB-6 and CPHB-7 transcripts accumulated in leaves during early stages of dehydration and decreased after prolonged dehydration. Both transcripts were also induced in ABA-treated callus. CPHB-3/4/5 were down-regulated by dehydration in both leaves and roots. The results support the role of HDZips in regulating programs of gene expression in C. plantagineum that lead to desiccation tolerance. PMID- 12081369 TI - A novel pollen-specific alpha-tubulin in sunflower: structure and characterization. AB - We describe here a new alpha-tubulin isoform from sunflower we named alpha(pi) tubulin. Alpha(pi)-tubulin is the most divergent higher-plant alpha-tubulin described so far, having an unusual deletion in the H1/B2 loop and a glutamine rich C-terminus. We constructed a three-dimensional model and discuss its implications. Using specific antibodies, we show that alpha(pi)-tubulin expression is restricted to the male gametophyte. Alpha(pi)-tubulin mRNA represents 90% of alpha-tubulin mRNA and a small percentage of total pollen mRNA. Among the plants tested, alpha(pi)-tubulin was only detected in sunflower and in Cosmos. Since both plants are Asteraceae, we propose that alpha(pi)-tubulin is specific to this family. Our results suggest that alpha(pi)-tubulin can inhibit tubulin assembly in pollen. This hypothesis is reinforced by the fact that alpha(pi)-tubulin is found in a complex with beta-tubulin in mature sunflower pollen. PMID- 12081370 TI - The DNA-compacting protein DCP68 from soybean chloroplasts is ferredoxin:sulfite reductase and co-localizes with the organellar nucleoid. AB - The multiple copies of the chloroplast genome (plastome) are condensed and organized into nucleoids by a set of proteins. One of these, the DNA-binding protein DCP68 from soybean, has previously been shown to compact DNA and to inhibit DNA synthesis in vitro. N-terminal amino acid analysis and the absorption spectrum of the purified protein suggest that DCP68 is the siroheme protein sulfite reductase, a ferredoxin-dependent enzyme that participates in sulfur assimilation for cysteine and methionine biosynthesis. The in vivo association of this protein with chloroplast nucleoids was confirmed by immuno-colocalization with antibodies against sulfite reductase from Arabidopsis thaliana. These results suggest that DCP68 is a bifunctional chloroplast protein that participates in reductive sulfur assimilation and plays a role in organellar nucleoid organization. The fact that dephosphorylation by alkaline phosphatase affects the binding of purified DCP68 to DNA in vitro might be indicative of the way the interaction of the protein with the nucleoid is regulated in vivo. PMID- 12081372 TI - Cell cycle-dependent and lysine residue-specific dynamic changes of histone H4 acetylation in barley. AB - Histone acetylation affects chromatin conformation and regulates various cellular functions, such as transcription and cell cycle progression. Although mitosis dependent transcriptional silencing and large-scale chromatin structural changes are well established, acetylation of histone H4 during the mitosis is poorly understood in plants. Here, the dynamics of acetylation of histone H4 in defined genome regions has been examined in the fixed barley cells throughout the mitosis by three-dimensional microscopy. Patterns of strong acetylation of the two lysine residues K5 and K16 of histone H4 in the barley genomes were found to be different. In interphase nuclei, H4 acetylated at K 16 was associated with the gene-rich, telomere-associated hemispheres, whereas K5 acetylation was detected in centromeric regions where the heterochromatin is distributed. Regions of strong K5 acetylation changed dynamically as the cell cycle proceeded. At prometaphase, centromeric acetylation at K5 decreased suddenly, with accompanying rapid increases of acetylation in the nucleolar organizing regions (NORs). Reverse changes occurred at telophase. On the other hand, the strongly acetylated regions of the K16 showed changes compatible with transcriptional activities and chromosome condensation throughout the cell cycle. Telomeric acetylation at K16 was detected throughout the cell cycle, although it was reduced at metaphase which corresponds to the most condensed stage of the chromosomes. It is concluded that dynamic changes in H4 acetylation occur in a lysine residue-, stage-, and region-specific manner and that they correlate with changes in the chromosome structure through the cell cycle. PMID- 12081371 TI - AtHVA22 gene family in Arabidopsis: phylogenetic relationship, ABA and stress regulation, and tissue-specific expression. AB - HVA22 is an ABA- and stress-inducible gene first isolated from barley (Hordeum vulgare L.). Homologues of HVA22 have been found in plants, animals, fungi and protozoa, but not in prokaryotes, suggesting that HVA22 plays a unique role in eukaryotes. Five HVA22 homologues, designated AtHVA22a, b, c, d and e, have been identified in Arabidopsis. These five AtHVA22 homologues can be separated into two subfamilies, with AtHVA22a, b and c grouped in one subfamily and AtHVA22d and e in the other. Phylogenetic analyses show that AtHVA22d and e are closer to barley HVA22 than to AtHVA22a, b and c, suggesting that the two subfamilies had diverged before the divergence of monocots and dicots. The distribution and size of exons of AtHVA22 homologues and barley HVA22 are similar, suggesting that these genes are descendents of a common ancestor. AtHVA22 homologues are differentially regulated by ABA, cold, dehydration and salt stresses. These four treatments enhance AtHVA22a, d and e expression, but have little or even suppressive effect on AtHVA22c expression. ABA and salt stress induce AtHVA22b expression, but cold stress suppresses ABA induction of this gene. Expression of AtHVA22d is the most tightly regulated by these four treatments among the five homologues. In general, AtHVA22 homologues are expressed at a higher level in flower buds and inflorescence stems than in rosette and cauline leaves. The expression level of these homologues in immature siliques is the lowest among all tissues analyzed. It is suggested that some of these AtHVA22 family members may play a role in stress tolerance, and others are involved in plant reproductive development. PMID- 12081373 TI - The upstream Sal repeat-containing segment of Arabidopsis thaliana ribosomal DNA intergenic region (IGR) enhances the activity of adjacent protein-coding genes. AB - The sequence containing 'upstream Sal repeats' (USR) from the Arabidopsis thaliana ribosomal DNA intergenic region (IGR) was tested for its influence on the in vivo activity of nearby protein coding genes. On average, the presence of the IGR fragment leads to a four-fold increase in the expression of a reporter gene, beta-glucuronidase, under control of the strong CaMV 35S promoter. With the help of the site-specific cre-lox recombination system, we have also obtained pairs of transgenic lines with or without the USR-containing fragment, both integrated at the same chromosomal position. Results with these transgenic lines, which contain an NPT II (kanamycin resistance) gene under control of the nos promoter as a test gene, confirmed the results obtained with the CaMV 35S-driven GUS gene. Moreover, they show that the IGR sequence can oppose tendencies of gene silencing. We hypothesize that the described effect relates to features of the chromatin structure in the proximity of the upstream Sal repeats. PMID- 12081374 TI - 5' deletion of a gbss1 promoter region from wheat leads to changes in tissue and developmental specificities. AB - Expression of granule-bound starch synthase 1 (GBSS1) in wheat is restricted to the grain filling process. In order to identify promoter regions which are involved in transcriptional control of the observed expression pattern, we isolated about 8 kb of a wheat gbss1-upstream region. Within this sequence several putative cis-acting elements were identified. In addition, an untranslated leader region is located in the 5' region of the gbss1 gene. To investigate promoter activity of the isolated region, the proximal 4.0 kb and progressively 5'-deleted fragments were transcriptionally fused to a beta glucuronidase reporter gene. The function of the promoter constructs was tested by transient expression assays in various wheat tissues and in transgenic wheat plants, which were selected for low number and integrity of transgene copies. Analysis of stable transformants revealed that the -4.0 kb promoter region mediates reporter gene expression that is in accordance with the endogenous gbss1 expression. Promoter deletion to -1.9 kb or to -1.0 kb did not change the expression profile with regard to grain and pollen specificity. However, the profile of beta-glucuronidase expression during the grain filling process is altered in such a way that the level of beta-glucuronidase activity declines due to the decreasing promoter length. It is proposed that enhancer elements and cis acting elements, which are involved in gbss1 transcription during the grain filling process, are located -1.9 kb upstream of the promoter. In addition, participation of the untranslated leader region in tissue-specific gene expression is discussed. PMID- 12081376 TI - Proceedings of the 2nd International Mental Lexicon Conference. Montreal, Canada, October 2000. PMID- 12081375 TI - Inducibility by pathogen attack and developmental regulation of the rice Ltp1 gene. AB - Using a genomic clone encoding a rice lipid transfer protein, LTP1, we analysed the activity of the 5' region of the Ltp1 gene in transgenic rice (Oryza sativa L.) during plant development and under pathogen attack. The -1176/+13, -556/+13 and -284/+13 regions of the promoter were fused upstream from the uidA reporter gene and nos 3' polyadenylation signal, resulting in the pdelta1176Gus, pdelta556Gus and pdelta284Gus constructs which were transferred to rice by microprojectile bombardment. Histochemical and fluorometric GUS assays and in situ detection of uidA transcripts in transgenic homozygous lines harbouring the pdelta1176Gus construct demonstrated that the Ltp1 promoter is preferentially active in aerial vegetative and reproductive organs and that both specificity and level of expression are regulated during organ development. In leaf sheath, GUS activity which is initially strictly localized in the epidermis of growing tissue, becomes restricted to the vascular system in mature tissues. In expanded leaf blade, expression of the uidA gene was restricted to the cutting level suggesting inducibility by wounding. Strong activity was detected in lemma and palea, sterile glumes, and immature anther walls and microspores but not in female reproductive organs. No GUS activity was detected during seed embryo maturation whereas the uidA gene was strongly expressed at early stages of somatic embryogenesis in scutellum tissue. The Ltp1 transcripts were found to strongly accumulate in response to inoculation with the fungal agent of the blast disease, Magnaporthe grisea, in two rice cultivars exhibiting compatible or incompatible host-pathogen interactions. Analysis of pdelta1176Gus leaf samples inoculated with the blast fungus demonstrated that the Ltp1 promoter is induced in all cell types of tissues surrounding the lesion and notably in stomata guard cells. In plants harbouring the Ltp1 promoter deletion construct pdelta556Gus, activity was solely detected in the vascular system of mature leaves whereas no uidA gene expression was observed in pdelta284Gus plants. These observations are consistent with the proposed role of LTP1 in strenghtening of structural barriers and organ protection against mechanical disruption and pathogen attack. PMID- 12081377 TI - Mental lexicon research in the new millennium. AB - This article presents an overview of current research on the mental lexicon, as it is represented by articles in the spring 2002 Special Issue of Brain and Language. We examine current findings in terms of language, task, and population effects associated with how words are presented and processed in the mind. We discuss how such mental representations may be linked to neurological instantiations and address the issue of ecological validity in the field. These discussions are organized in order to both provide an overview of the issues and to enable the reader to locate specific articles that bear on these issues. Finally, we present an organizational framework for the characterization of mental lexicon research within which challenges for advancement are isolated. PMID- 12081379 TI - Does prime modality influence morphological processing? AB - Facilitation among morphologically related words generally is impervious to the prefixed or suffixed structure of primes and targets. A notable exception arises, however, when both primes and targets are suffixed. More specifically, when primes are auditory and targets are visual, facilitation for a suffixed target (e.g., payment) is absent when it follows a prime (e.g., payable) that is morphologically related and suffixed (Marslen-Wilson, Tyler, Waksler, & Older, 1994). To account for null facilitation (viz., the "suffix-suffix" effect), Marslen-Wilson and his colleagues posit inhibitory links between suffixes of morphological relatives. The present study assesses the generality of the "suffix suffix" effect. When morphological facilitation is assessed relative to an orthographically related baseline, suffixed primes facilitate derivationally as well as inflectionally related morphological targets when primes are visual as well as auditory in both the lexical decision and naming tasks. The present findings call into question lexical models that posit inhibitory links between suffixes of morphological relatives. PMID- 12081378 TI - Graded aspects of morphological processing: task and processing time. AB - Effects on targets of orthographically (O) and semantically (S) related primes were compared with morphologically related (M) primes in the lexical decision, naming, and go/no go naming tasks. The overall pattern typified the graded nature of morphological processing. Morphological relatedness produced facilitation whose magnitude varied across a range of stimulus onset asynchronies (SOAs of 66 300 ms) and tasks. The effect of semantic and orthographic similarity also depended on SOA and on task. Importantly, the effects of morphological relatedness and orthographic similarity diverged along a time course that reflected semantic processing but could only be approximated by the effect of semantic relatedness between prime and target. PMID- 12081380 TI - Recall of morphologically complex forms is affected by memory task but not dyslexia. AB - The authors studied the effect of morphological complexity on working memory in list recall tasks with base words (boy), inflected words (boy + 's) and derived words (boy + hood) in a morphologically rich language: Finnish. Simple serial recall was compared to complex working memory tasks, combining word recall with sentence verification in 8-year-old normally reading participants, dyslexic children, and adults. The normally reading children performed better than dyslexic children on both memory tasks and a test of morphology. Base words were better recalled than morphologically complex words. Memory was better for derived than inflected words in simple but not complex span tasks. There was no interaction between word type and reading group and thus no suggestion of dyslexia being associated with specific problems to represent complex morphology in working memory. Morphological processing in working memory appeared to depend on the task. PMID- 12081381 TI - The subjects as a simple random effect fallacy: subject variability and morphological family effects in the mental lexicon. AB - This is a methodological study addressing the appropriateness of standard by subject and by-item averaging procedures for the analysis of repeated-measures designs. By means of a reanalysis of published data (Schreuder & Baayen, 1997), using random regression models, we present a proof of existence of systematic variability between participants that is ignored in the standard psycholinguistic analytical procedures. By applying linear mixed effects modeling (Pinheiro & Bates, 2000), we call attention to the potential lack of power of the by-subject and by-item analyses, which in this case study fail to reveal the coexistence of a facilitatory family size effect and an inhibitory family frequency effect in visual and auditory lexical processing. PMID- 12081382 TI - The probability of the least likely non-length-controlled bigram affects lexical decision reaction times. AB - The frequency effect, by which high frequency words are recognized with more ease than low frequency words, is one of the most robust effects in cognitive psychology. Frequency interacts with many word-level variables, to the extent that most effects reported in word recognition literature have an impact only on low frequency words. This has been taken as evidence that high frequency words are accessed in a special way, via either an addressed pathway as in the dual route model or an assembled pathway as in a PDP model. Under either model, sublexical effects should have no bearing on the ease with which representations for high frequency words are accessed. In this article, however, we describe a series of studies that examine a sublexical effect (namely nonlength controlled minimal bigram frequency) that is only found for high frequency words, suggesting that sublexical processing must play a role in the recognition of even high frequency words. PMID- 12081383 TI - Stem homographs and lemma level representations. AB - Previous studies have found that priming a target like cerr-o ("hill") with an unmasked stem-homograph (e.g., cerr-ar, "to close") or with an unmasked allomorph of a stem-homograph (e.g., cierr-a, "he/she/it closes") has an inhibitory effect on target recognition. We report evidence from stem-homograph priming studies that implicate both morphological parsing and lexical selection processes at the lemma level. We argue that stem-homograph inhibition is the product of lexical selection mechanisms that are engaged just when a fully ambiguous stimulus (here, an inflectional stem) must be assigned one of its viable interpretations in order to afford conscious perception of lexical identity. PMID- 12081384 TI - Cross-modal morphological priming in French. AB - We investigated the lexical representation of morphologically complex words in French using a cross-modal priming experiment. We asked if the lexical representation for derivationally suffixed and prefixed words is morphologically structured and how this relates to the phonological transparency of the surface relationship between stem and affix. Overall we observed a clear effect of the morphological structure for derived words, an effect that is not explicable by a formal effect. Prefixed words prime their stems, even when they have a phonologically opaque relationship, and a prefixed word primes another prefixed word derived from the same stem. However, suffixed words prime their stems only if their relationship is phonologically transparent. Two suffixed words derived from the same stem prime each other. These two latter results differ from those observed in English by Marslen-Wilson, Tyler, Waksler, and Older (1994). We argue that it is the specific properties of the language, such as rhythm, that could explain the differences between the results observed for the two languages and we propose a model where prefixed and suffixed words are decomposed at different stages during their identification process. PMID- 12081385 TI - Regularity revisited: evidence from lexical access of verbs and nouns in Greek. AB - The present study investigates the relationship between morphological regularity and form during lexical processing using a visual priming paradigm varying the prime duration. We addressed the effect of regularity on morphological facilitation in nouns and verbs by exploiting particular characteristics of a highly inflected language, Greek, in which it is possible to manipulate morphological regularity while controlling the degree of orthographic overlap between morphological relatives. The effects of morphological regularity were found to crucially depend on the time course of lexical access. Moreover, morphological regularity was found to affect nouns and verbs differentially. We interpret these findings with respect to the distinction between affix processing and allomorph retrieval and discuss the issues of form overlap and orthographic boundaries in morphological processing. PMID- 12081386 TI - Rapid activation of the lexicon: a further investigation with behavioral and computational results. AB - Priming effects were observed in a categorization task for both prime-target synonym pairs (e.g., boat-ship) and first-associate pairs (e.g., boat-sea). However, the amount and onset of priming were different for synonyms and associated pairs. The effect appeared sooner for synonyms (at prime duration of 43 ms) than for associated words (57 ms onset) but was present for both these relationships at 71 ms of prime presentation. A prime visibility pretest was conducted with the same participants in order to determine the rate of recognition of our prime words. Last, a French matrix of the HAL model was built, which showed that synonyms pairs were semantically closer than associated pairs. These results are in accordance with our previous study (Frenck-Mestre & Bueno, 1999) and are discussed in relation with semantic models, such as Plaut's (1995) distributed model. PMID- 12081387 TI - Hemispheric contributions to lexical ambiguity resolution: evidence from individuals with complex language impairment following left-hemisphere lesions. AB - Nine individuals with complex language deficits following left-hemisphere cortical lesions and a matched control group (n = 9) performed speeded lexical decisions on the third word of auditory word triplets containing a lexical ambiguity. The critical conditions were concordant (e.g., coin-bank-money), discordant (e.g., river-bank-money), neutral (e.g., day-bank-money), and unrelated (e.g., river-day-money). Triplets were presented with an interstimulus interval (ISI) of 100 and 1250 ms. Overall, the left-hemisphere-damaged subjects appeared able to exhaustively access meanings for lexical ambiguities rapidly, but were unable to reduce the level of activation for contextually inappropriate meanings at both short and long ISIs, unlike control subjects. These findings are consistent with a disruption of the proposed role of the left hemisphere in selecting and suppressing meanings via contextual integration and a sparing of the right-hemisphere mechanisms responsible for maintaining alternative meanings. PMID- 12081389 TI - The recognition of reduced word forms. AB - This article addresses the recognition of reduced word forms, which are frequent in casual speech. We describe two experiments on Dutch showing that listeners only recognize highly reduced forms well when these forms are presented in their full context and that the probability that a listener recognizes a word form in limited context is strongly correlated with the degree of reduction of the form. Moreover, we show that the effect of degree of reduction can only partly be interpreted as the effect of the intelligibility of the acoustic signal, which is negatively correlated with degree of reduction. We discuss the consequences of our findings for models of spoken word recognition and especially for the role that storage plays in these models. PMID- 12081388 TI - The role of the syllable in lexical segmentation in French: word-spotting data. AB - Three word-spotting experiments assessed the role of syllable onsets and offsets in lexical segmentation. Participants detected CVC words embedded initially or finally in bisyllabic nonwords with aligned (CVC.CVC) or misaligned (CV.CCVC) syllabic structure. A misalignment between word and syllable onsets (Experiment 1) produced a greater perceptual cost than a misalignment between word and syllable offsets (Experiments 2 and 3). These results suggest that listeners rely on syllable onsets to locate the beginning of words. The implications for theories of lexical access in continuous speech are discussed. PMID- 12081390 TI - Ageing and lexical access to common and proper names in picture naming. AB - The question of whether lexical access for proper names is more impaired by ageing than lexical acess for other words is controversial. The aim of the present investigation was to evaluate the effect of age on proper and common name retrieval in long-term memory. The word retrieval paradigm used to achieve this goal consisted of the naming of photographs representing celebrities (production of a certain kind of proper names: names of people) and objects (production of common names). Compared with younger adults, elderly people experienced more tip of-the-tongue states for proper names, but not for common names. Thus, the present study provides support for disproportionate age-related problems in lexical access to proper names. This finding is interpreted with reference to the cognitive model of speech production proposed by Burke et al. (1991). PMID- 12081391 TI - How are inflectional affixes organized in the mental lexicon?: evidence from the investigation of agreement errors in agrammatic aphasics. AB - Recent psycholinguistic studies have provided evidence that regularly inflected words are decomposed into stems and affixes, both of which have their own representations in the mental lexicon. Specific models of the lexical organization of inflectional affixes have, however, only rarely been investigated in psycho- or neurolinguistic work. We test two recently proposed theoretical models: a representation of affixes (i) in default inheritance trees (Corbett and Fraser, 1993) and (ii) in underspecified paradigms (Wunderlich, 1996). Based on an analysis of agreement errors in elicited speech-production data of German agrammatic aphasics, we argue that affixes are organized with respect to the morphosyntactic features they encode. Specifically, our data indicate that inflectional affixes are best captured within an underspecified paradigm. PMID- 12081392 TI - English derivational suffix frequency and children's stress judgments. AB - Considering the importance of word and stem frequency in the adult lexical processing literature, and the effect of input frequency on children's acquisition of words (Tardif, Shatz, and Naigles, 1997), it was hypothesized that children's acquisition of English morphologically conditioned stress alternations would be affected by the frequency with which children were exposed to different stress-changing suffixes (e.g., -tion, -ity, and -ic). Study 1 determined the proportional representation of suffixes in a children's literature corpus, thereby allowing the suffix variable to be established. Study 2 empirically examined the effect of suffix frequency on school-aged children's judgments of primary stress placement. Findings suggest that age and suffix frequency both play a role in children's awareness of stress placement. PMID- 12081393 TI - The processing of lexical ambiguity: homonymy and polysemy in the mental lexicon. AB - Under the theoretical assumption that lexical ambiguity is not a homogeneous phenomenon, but rather that it is subdivided into two distinct types, namely homonymy and polysemy, the present study investigated whether these different types of lexical ambiguity are psychologically real. Four types of ambiguous words, homonymous words (e.g., "pen"), polysemous words with metaphorical extensions (e.g., "eye"), polysemous words with a count/mass metonymic extension (e.g., "turkey"), and polysemous words with a producer/product metonymic extension (e.g., "Dali"), were used in a cross-modal sentence-priming lexical decision task. Overall, the theoretical distinction between homonymy and polysemy was reflected in the results of the present study, which revealed differential processing depending on the type of ambiguity. PMID- 12081394 TI - Class specificity and the lexical encoding of participant information. AB - It is commonly assumed across the language sciences that some semantic participant information is lexically encoded and some is not. In this article, we propose that semantic obligatoriness and verb class specificity are criteria which influence whether semantic information is lexically encoded. We present a comprehensive survey of the English verbal lexicon and two continuation studies which confirm that both factors play a role in the lexical encoding of participant information. PMID- 12081395 TI - Grammatical gender in the production of single words: some evidence from Greek. AB - The present study investigated the effect of prior grammatical gender information provided by the production of a bare noun on the production of a syntactically unrelated, gender-inflected color adjective. The target language was Greek. A lexical priming task involving picture naming was employed. Participants saw a series of pictures, some in color and some in black-and-white; they had to name a black-and-white picture with the single noun (prime) and a color picture with the appropriately inflected color adjective (target). Prior gender information was shown to affect subsequent production of gender-marked words. The effect was restricted to nouns of one gender class (masculine) only. The implications of these results for the representation and processing of gender in production are discussed. PMID- 12081396 TI - Derivation by prefixation in Slovenian: two aphasia case studies. AB - The present study is concerned with two Slovenian-speaking patients who were asked to produce, in various tasks, verbs, nouns, and adjectives derived by prefixation with prepositions. Despite differences due to their specific linguistic difficulties, both patients' performance was characterized by the differential processing of prefixes and remaining components of complex words. Prepositions in prefixation were mostly preserved, and less frequently substituted, regardless of the numerous errors produced in the remaining portion of the words. These errors seem clearly determined by the morphological structure of the words and therefore appear to be authentic morphological errors. These findings contribute to the theoretical debate on mental lexical representation, speaking in favor of a morphological decomposition in processing of prefixed complex words at different processing levels. PMID- 12081397 TI - Lexical representations of written nouns and verbs in Italian. AB - This study investigates the differences in the input representations of verbs and nouns in the Italian mental lexicon. One visual lexical decision experiment in which the stem homograph effect (Laudanna, Badecker, and Caramazza, 1989) was evaluated on noun and verb targets is reported. Strong inhibitory effects on verb targets preceded by stem homographs relative to nonhomographic word pairs were found. The effects on verb targets were also much more reliable than the effects on noun targets. The pattern of results provides support for the hypothesis that noun and verbs are differently represented in the orthographic input lexicon. PMID- 12081398 TI - The retrieval of syntactic and semantic information in the production of verbs. AB - In Italian, there are two aspectual auxiliaries: essere (to be) and avere (to have). According to the syntactic hypothesis, aspectual auxiliary assignment in Italian is a syntactic phenomenon. Using the picture-word interference paradigm, Experiment 1 tested the predictions of this view. Results failed to support the syntactic hypothesis but could be explained within a semantic framework: A verb's thematic grid is involved in aspectual auxiliary assignment. Experiment 2 tested this hypothesis. The findings corroborated the predictions. The evidence is interpreted as suggesting the involvement of thematic information in the assignment of the aspectual auxiliary during the production of Italian verbs. PMID- 12081399 TI - Processing and Representation of German -n Plurals: A Dual Mechanism Approach. AB - Four subclasses of German noun plurals affixed with -n, exhibiting different degrees of predictability, are investigated in a lexical decision experiment with visual stimuli and in a cross-modal priming experiment. The lexical decision experiment compared -n plurals and irregular -er plurals to plurals with the default affix -s. Whereas the -s plurals yielded no word-form frequency effects, such effects could be observed for all other plural forms, including all subclasses of -n plurals. The priming experiment produced a different pattern. We found that the prior presentations of fully predictable -n plurals prime the stems as effectively as the stem itself, whereas -n plurals with limited predictability yielded only reduced priming effects toward their stems. These results are explained in terms of an extended dual mechanism approach, taking into account an access level and a central level of lexical processing. PMID- 12081400 TI - Dissociation in Italian conjugations: a single-route account. AB - Say and Clahsen (S and C) report an experiment involving assignment of past participle suffixes to nonce words in Italian. Their evidence suggests a dual route model that assigns the theme vowel of the 1st conjugation, while storing it lexically in other conjugations. However, these nonce words were assigned suffixes by a computer algorithm that determined the past participle of the nonce items on the basis of phonological similarity to existing inflected forms. The outcome of the simulation mirrors that of the subjects closely, suggesting that the dissociations found by S and C are not adequate evidence for a dual-route model of Italian stem formation. PMID- 12081401 TI - German noun plurals: a challenge to the dual-mechanism model. AB - In this article, the authors test one of the central claims of the Dual-Mechanism Model (Pinker and Prince, 1994), that is, that regular inflection equals default inflection. Based on results from an elicitation task with eight agrammatic Broca's aphasics and a lexical decision task with unimpaired subjects, the authors show that this assumption is not borne out. Their data on German plural inflection rather indicate that regular inflection is not necessarily identical to default inflection. To capture the German data, they have to assume regular but input-restricted inflection besides regular default inflection. PMID- 12081402 TI - The contribution of frequency and semantic similarity to morphological processing. AB - This research investigated the role of frequency and semantic factors in the processing of morphologically complex words. Unaffixed targets were immediately preceded by either inflected or derived primes that were either high or low in frequency. In Experiment 1, low-frequency inflections and derivations produced equivalent priming effects, whereas high-frequency inflections produced more priming than derivations. In Experiment 2, both semantic and morphological priming varied as a function of SOA (50, 150, and 250 ms). The difference in priming of inflections and derivations increased as SOA increased. Results suggest that frequency and semantic similarity interactively modulate the magnitude of morphological priming and, by inference, affect the processing of morphologically complex words. Dual-route and connectionist accounts are discussed. PMID- 12081403 TI - Identity avoidance in the Hebrew lexicon: implications for symbolic accounts of word formation. AB - Hebrew frequently exhibits geminates in the root but strictly constrains their location: Root-initial gemination is rare (e.g., ssm), whereas root-final gemination (e.g., smm) is frequent. Four experiments demonstrate that Hebrew speakers generalize this constraint to novel roots. When speakers are encouraged to form a triliteral root from a biconsonantal input (e.g., sm), they frequently reduplicate the root's final radical (e.g., smm), but not its initial radical (e.g., ssm). Likewise, the rejection of novel root foils with root initial geminates is easier than roots with final geminates. In both cases, speakers' performance is inexplicable by the statistical structure of the Hebrew language. Speakers' ability to freely generalize the constraint on root structure suggests that their linguistic competence appeals to mental variables. PMID- 12081404 TI - The selection of grammatical features in word production: the case of plural nouns in German. AB - Two experiments investigate the effect of number congruency using picture-word interference. Native German participants were required to name pictures of single objects (Nase 'nose') or two instances of the same object (Nasen 'noses') while ignoring simultaneously presented distractor words. Distractor words either had the same number or were different in number. In addition, the type of plural formation (same or different inflectional plural suffix) and the semantic relationship (same or different semantic category) between target and distractor were varied in Experiments 1 and 2. Results showed no effect of number congruency in either experiment. Furthermore, the type of inflectional suffix did not exert an influence on naming latencies in Experiment 1, but semantic relationship led to a significant interference effect in Experiment 2. The results indicate that selection of the number feature diacritic in noun production is not a competitive process. The implications of the results for models of lexical access are discussed. PMID- 12081405 TI - Where and how morphologically complex words interplay with naming pictures. AB - Two picture-word experiments are reported in which a delay of 7 to 10 was introduced between distractor and picture. Distractor words were either derived words (Experiment 1) or compounds (Experiment 2), morphologically related to the picture name. In both experiments, the position of morphological overlap between distractor (e.g., rosebud vs tea-rose) and picture name (rose) was manipulated. Clear facilitation of picture naming latencies was obtained when pictures were paired with morphological distractors, and effects were independent of distractor type and position of overlap. The results are evaluated against "full listing" and "decomposition" approaches of morphological representation. PMID- 12081406 TI - Sonority constraints on onset-rime cohesion: evidence from native and bilingual Filipino readers of English. AB - Research in English suggests that syllables can be analyzed in terms of two subunits-the onset (defined as the initial consonant or consonant cluster) and the rime (the unit formed by the vowel and following consonant/s). This study investigated whether nonnative readers of English, which in the case of the present study were native Filipino speakers, also make use of onset-rime units, particularly when some features of their native language (namely infixation and reduplication) appear to foster no awareness of such units. In two lexical decision experiments, monosyllabic English words were presented, divided in between their first and second consonants (e.g., B LIND), at their onset-rime boundary (e.g., BL IND), or at their antibody boundary (e.g., BLI ND). Results indicated that the processes of infixation and reduplication did not affect the English word processing of native Filipino speakers. Rather, results for both native Filipino and native English speakers suggest that onsets composed of an "s + consonant" sequence (e.g., STAMP) are less cohesive than onsets comprised of a stop-liquid sequence (e.g., BLIND). It was concluded that not only may sonority constraints underlie onset cohesiveness, but that such phonetic properties may also be involved in visual word recognition. PMID- 12081407 TI - Is phonological information mapped onto semantic information in a one-to-one manner? AB - Spoken word recognition models have to explain the influence of mismatching information on lexical activation. The effect of mismatching information is usually addressed with cross-modal semantic priming experiments using priming effects as a measure of the degree of lexical activation. Pseudowords phonologically related to a semantic associate of the target, e.g., *domato PAPRIKA, serve as primes. Mismatch effects at the word form level are supposed to percolate unaltered to the semantic level. We show that cross-modal semantic priming might underestimate activation at the word form level. Targets (e.g., PAPRIKA) were preceded by either phonologically related pseudoword primes (e.g., *baprika) or semantically related pseudoword primes (e.g., *domato). Different priming and RT patterns were obtained for the two priming relations. PMID- 12081408 TI - The influence of age of acquisition, root frequency, and context availability in processing nouns and verbs. AB - In the Italian language there is a higher number of inflectional suffixes in verbs than in nouns, and this might imply that verbs are more likely to undergo a morphological analysis in terms of root and suffix as compared to nouns (Traficante & Burani, unpublished observations). Moreover, verbs tend to be more abstract than nouns, and this aspect might make verb processing more difficult. Finally, the developmental gap in the production of nouns and verbs suggests that age of acquisition might affect noun and verbs differently. Nouns and verbs were presented in a lexical decision and in a naming task. The morphological variable root frequency in addition to word frequency, length and word age of acquisition, and the semantic variables concreteness and context availability (Schwanenflugel, Harnishfeger, & Stowe, 1988) were used as predictors in multiple-regression analyses in which lexical decision and naming latencies were the dependent variables. The results showed that age of acquisition, context availability, and root frequency are all important in predicting both lexical decision and naming latencies for nouns and verbs, but age of acquisition and root frequency are better predictors of the differences in processing Italian nouns and verbs. PMID- 12081409 TI - Form-based representation in the mental lexicon: priming (with) bound stem allomorphs in Finnish. AB - A series of four visual-visual priming experiments investigates the role of bound stem allomorphs in the representation and processing of Finnish case inflected nouns. Niemi et al. (1994) and Laine et al. (1994) argue that Finnish nouns are parsed into stem and affix in reception and that the bound stem allomorphs have separate (visual) lexical representations. Recently Jarvikivi and Niemi (in press) have provided converging evidence for their claim based on a series of lexical decision experiments with Finnish stem allomorphs. The results from the present series of four follow-up experiments reported here showed that (isolated) bound stem allomorphs primed the recognition of the corresponding monomorphemic nouns significantly compared both to phonologically unrelated pseudowords and to phonologically minimally different pseudowords. Furthermore, not only did both phonologically transparent and opaque case inflected nouns prime the corresponding nominative singulars, but also there was no difference in priming between the two. The results are well in accordance and corroborate the hypotheses drawn from the earlier investigations. Moreover, they further indicate that bound stem allomorphs have representations on a purely formal level only, i.e., they serve as indices of and entering points to morphological/morphosyntactic information. PMID- 12081410 TI - Experimental evidence for serial models of lexical access: a judgment task. AB - The hypothesis that the lemma and lexeme levels of lexical access are in a feed forward serial relation is supported. Subjects judge the degree of semantic relatedness of pairs of words. Stimuli are all functionally synonymous; they are pairs of words implicated in natural word blends. Half the stimuli are phonologically related and the other half are not. In one experiment the pairs are presented in a sentential context and in the other no context is provided. In both the experiments the phonologically unrelated pairs are judged closer in meaning than the phonologically related pairs. This is interpreted as support for serial models of lexical access. PMID- 12081411 TI - Verb-noun double dissociation in aphasic lexical impairments: the role of word frequency and imageability. AB - Neurolinguistic studies have provided important evidence regarding the organization of lexical representations and the structure of underlying conceptual knowledge; in particular, it has been shown that the retrieval of verbs and nouns can be damaged selectively. Dissociated lexical damage is proof of an independent mental organization of lexical representations and/or of the underlying processes. The aim of the present study is to estimate the rate of dissociated impairments for nouns and verbs on a large sample of mild to moderate aphasic patients and to investigate the mechanisms underlying such phenomena. In addition, the authors wished to verify to what degree the impairment for nouns and verbs is related to a specific type of language disorder. A confrontation naming task for verbs and nouns was administered to 58 aphasic patients. The major lexical (word frequency and age of acquisition) and semantic variables (familiarity and imageability of the underlying concept) were considered for each noun and verb used in the task. Verbs were distinguished by major functional classes (transitive, intransitive, and ergative verbs). The data collected from this task were analyzed twice: (i) as a group study comparison of major aphasic subgroups and (ii) as a multiple single case study to evaluate the differences on the naming of verbs and nouns and the effect of the lexical semantic variables on each individual patient. The results confirm the existence of dissociated naming impairments of verbs and nouns. Selective impairment of verbs is more frequent (34%) than that of nouns (10%). In many cases, the dissociated pattern of naming impairment disappeared when the effect of the concomitant variables (word frequency and imageability) was removed, but in approximately one-fifth of the cases the noun or verb superiority was preserved. Noun superiority emerged in five of six agrammatic patients. Both the naming of verbs (n = 9) or of nouns (n = 6) could be impaired selectively in fluent aphasic patients. The results lend support to the hypothesis of an independent mental organization of nouns and verbs, but a substantial effect of imageability and word frequency suggests an interaction of the naming impairment with underlying lexical and semantic aspects. PMID- 12081412 TI - Interactive processing of phonological information in reading Japanese Kanji character words and their phonemic radicals. AB - Kanji are categorized into four types based on the combinations of "subword validity" (when the right phonemic radical represents the same On-reading as the whole Kanji character) and "radical-neighbor consistency" (when the whole Kanji character represents the same On-reading as all of its neighbor characters). The study demonstrated that both subword validity and radical-neighbor consistency affect naming latencies and error rates regardless of character frequency. The study also demonstrated that the subword validity affects ease of extraction of a subword's phonology. These results suggest that the phonology of the whole Kanji competes with the subword's phonology. Moreover, the competition is stronger when the radical-neighbors aid in extracting the phonology of the whole Kanji. PMID- 12081413 TI - Whole word morphologizer: expanding the word-based lexicon: a nonstochastic computational approach. AB - Whole Word Morphologizer is a small computer implementation of word-based morphology. The program automatically identifies morphological relations in a small word-based lexicon, literally learning its morphology, and uses the knowledge it acquires to generate new words. It is based on a model of the mental lexicon in which all entries are whole, entire, fully fledged words and relies solely on basic cognitive principles (differentiation and generalization) for the automatic acquisition of morphological relations and the population of the lexicon. PMID- 12081414 TI - Schemas and competing paradigms in Swedish plural formation. AB - According to autonomous descriptions of Swedish morphology, for the majority of nouns the gender of the lexeme and the phonological nature of the last segment or segments of the base form (indefinite singular) determine the inflectional category of the noun. The purpose of the present study is to find out if Swedish may resort to the use of larger shapes than the terminal segment or segments in the allegedly ambiguous monosyllabic CVC words. It has been shown in other languages-here giving English as an example-that items in semiproductive classes (like the English ablaut verbs string:strung and spin:spun) are associated via formal phonological links or bundles of links that constrain the manner in which native speakers pattern these form-to-form connections within morphological paradigms (see Bybee & Slobin, 1982; Bybee, 1985). The present wug-type experiment with legal nonwords was administered to three groups of children, i.e., 8- to 9-year-olds, 10- to 11-year-olds, and 15-year-olds. It was hypothesized that the ambiguous monosyllabic CVC lexemes would stabilize themselves slowly and late during language acquisition. The results show a vowel and consonant effect in the en-gender CVC nonwords. Swedish thus resorts to the use of larger shapes (schema analogies) than the terminal segment or segments. Moreover, a development of paradigm membership toward adultlike patterns is seen at late stages of language acquisition of both the ambiguous and the unambiguous nonword types. PMID- 12081415 TI - Morphological representation in an endangered, polysynthetic language. AB - This article presents the results from an initial psycholinguistic study of patterns of morphological representation in Dene Suline (Chipewyan), an indigenous and highly endangered language spoken in Northwestern Canada. Our investigation focused on how morphological knowledge in this polysynthetic language is affected across various levels of language attrition by employing a morphological segmentation task and an off-line lexical decision task. We discuss the manner in which these tasks target different aspects of morphological ability and then turn to methodological issues associated with conducting psycholinguistic studies with language users that differ in levels of age, education, literacy, and bilingualism (Dene- English). Finally, we report on the challenges of doing psycholinguistic research outside the confines of a university setting and make some recommendations to other researchers who might wish to undertake similar studies. PMID- 12081416 TI - Auditory verb recognition in developmental language impairment. AB - This research addresses the issues of frequency and regularity in auditory verb recognition and their importance for the organization of the mental lexicon in DLI francophones. A primed auditory lexical decision task probes reaction times and response accuracy in DLI and control participants on frequent and infrequent, and regular and irregular, inflected forms of verbs. Results show no priming for inflectionally related forms as well as strong frequency effects in DLI participants. These findings lend additional support to the hypothesis that developmental language impairment is a deficit in lexical representation. PMID- 12081417 TI - The effect of number of kanji radical companions in character activation with a multi-radical-display task. AB - The majority of Japanese complex kanji consist of two subword units (radicals), and each radical is differentiated according to its combinability. Radical-type frequency is defined as the number of radical companions (NC) functioning as the number of characters containing that radical. The current study is intended to address the question of whether the NC plays a role in kanji character activation under a multi-radical-display (MRD) instead of a tachistoscopic display. An MRD task was used in which the NC was manipulated and the frequency of use of the whole characters was matched on average across the experimental conditions. Participants were divided into two groups (good knowledge vs poor knowledge) according to the score of a kanji completion task. The participants' task was to select an appropriate combination of radicals to compose a legitimate character under the MRD condition. The poor-knowledge group yielded relatively more errors than did the good-knowledge group in the low-NC condition, whereas the good knowledge group yielded more errors in the high-NC condition than in the low-NC condition. These results demonstrate the different influences of NC as metaknowledge (tacit knowledge of radical productivity) in both good and poor readers. PMID- 12081418 TI - Activation of phonological codes during reading: evidence from errors detection and eye movements. AB - This experiment explored the role of phonology in the activation of word meanings when homophonic and nonhomophonic errors were embedded in meaningful texts. The resulting data supported the position that phonological codes are activated very early in eye fixation and are compatible with the verification model of Van Orden (1987). PMID- 12081419 TI - Conceptual representation of verbs in bilinguals: semantic field effects and a second-language performance paradox. AB - It is well known that bilinguals perform better in their first language (L1) than in their second lanaguage (L2) in a wide range of linguistic tasks. In recent studies, however, the authors have found that bilingual participants can demonstrate faster response times to L1 stimuli than to L2 stimuli in one classification task and the reverse in a different classification task. In the current study, they investigated the reasons for this "L2-better-than-L1" effect. English-French bilinguals performed one word relatedness and two categorization tasks with verbs of motion (e.g., run) and psychological verbs (e.g., admire) in both languages. In the word relatedness task, participants judged how closely related pairs of verbs from both categories were. In a speeded semantic categorization task, participants classified the verbs according to their semantic category (psychological or motion). In an arbitrary classification task, participants had to learn how verbs had been assigned to two arbitrary categories. Participants performed better in L1 in the semantic classification task but paradoxically better in L2 in the arbitrary classification task. To account for these effects, the authors used the ratings from the word relatedness task to plot three-dimensional "semantic fields" for the verbs. Cross-language field differences were found to be significantly related to the paradoxical performance and to fluency levels. The results have implications for understanding of how bilinguals represent verbs in the mental lexicon. PMID- 12081421 TI - Homophonic forms of regularly inflected verbs have their own orthographic representations: a developmental perspective on spelling errors. AB - In previous research (Sandra, Frisson, & Daems, 1999) we demonstrated that experienced writers of Dutch (18-year-olds) make spelling errors on regularly inflected homophonic verb forms. Intrusion errors, i.e., spelling of the homophonic alternative, occurred more often when the low-frequency homophone had to be written. In the present article we report error data for three groups of less experienced spellers, who have not yet fully mastered the rules for verb suffix spelling: 12-year-olds, 13-year-olds, and 14-year-olds. Younger spellers obviously make many more errors than experienced ones. Whereas this is in part due to inadequate rule mastery/application, their error patterns are also clearly influenced by the frequency relationship between the homophonic forms, i.e., the same factor accounting for the errors of experienced spellers. The conclusion of our present and past research is that homophonic forms of regularly inflected verbs have their own orthographic representations in the mental lexicon and that these representations cause interference in writing (spelling errors), whereas they might cause facilitation in reading (a claim made by dual-route models of reading). PMID- 12081420 TI - Orthographic processing of polysyllabic words by native and nonnative English speakers. AB - How polysyllabic English words are analyzed in silent reading was examined in three experiments by comparing lexical decision responses to words physically split on the screen. The gap was compatible either with the Maximal Onset Principle or the Maximal Coda Principle. The former corresponds to the spoken syllable (e.g., ca det), except when the word has a stressed short first vowel (e.g., ra dish), while the reverse is true for the latter (giving cad et and rad ish). Native English speakers demonstrated a general preference for the Max Coda analysis and a correlation with reading ability when such an analysis did not correspond with the spoken syllable. Native Japanese speakers, on the other hand, showed a Max Onset preference regardless of the type of word, while native Mandarin Chinese speakers showed no preference at all. It is concluded that a maximization of the coda is the optimal representation of polysyllabic words in English and that poorer native readers are more influenced by phonology than are better readers. The way that nonnative readers mentally represent polysyllabic English words is affected by the way such words are structured in their native language, which may not lead to optimal English processing. PMID- 12081422 TI - The processing and representation of Dutch and English compounds: peripheral morphological and central orthographic effects. AB - In this study, we use the association between various measures of the morphological family and decision latencies to reveal the way in which the components of Dutch and English compounds are processed. The results show that for constituents of concatenated compounds in both languages, a position-related token count of the morphological family plays a role, whereas English open compounds show an effect of a type count, similar to the effect of family size for simplex words. When Dutch compounds are written with an artificial space, they reveal no effect of type count, which shows that the differential effect for the English open compounds is not superficial. The final experiment provides converging evidence for the lexical consequences of the space in English compounds. Decision latencies for English simplex words are better predicted from counts of the morphological family that include concatenated and hyphenated but not open family members. PMID- 12081423 TI - How early does morpholexical reading develop in readers of a shallow orthography? AB - In this research, lexical and morpholexical reading in Italian children ages 8 to 10 years were investigated. Children and control adults were administered two tasks on words and pseudowords: visual lexical decision and naming. Word frequency effects in both lexical decision and naming were found in both children and adults. For all age groups pseudowords made up of roots and derivational suffixes were decided more frequently as possible words and were named more quickly and accurately than matched pseudowords with no morphological constituency. These results show that morpholexical reading is available and efficient in young readers of a shallow orthography, with similar patterns in children and adults. PMID- 12081424 TI - Masked priming of prefixes and the influence of spelling-meaning consistency. AB - Two experiments are reported which examined whether a prefixed word can be primed by a word sharing only the prefix letters in a masked priming paradigm. In addition the studies examined whether the size of the priming effect is influenced by the consistency with which a prefix letter pattern appears in real prefixed words. The ratio of real prefixed words to all words containing a prefix letter pattern was calculated and used to identify high-consistency prefixes, which are frequently used as prefixes (e.g., UN in UNHAPPY) and rarely appear as pseudoprefixes (e.g., UNCLE), and low-consistency prefixes, which appear in many pseudoprefixed words (e.g., DE in DESERT) but fewer truly prefixed words (e.g., DECODE). In Experiment 1, decision latencies for both types of prefixes were facilitated when real prefixed target words were preceded by real prefixed prime words in a short SOA masked priming paradigm, although the size of the priming effect for low-consistency prefixes was similar in size to that for orthographic controls. In Experiment 2, real prefixed target words were preceded by pseudoprefixed prime words. Facilitation in performance remained for high consistency prefixes but was absent for low-consistency prefixes. These results support models of morphological processing that are sensitive to the statistical nature of the relationships between orthographic and semantic representations in a language. PMID- 12081425 TI - Selective impairment of lexical stress assignment in an Italian-speaking aphasic patient. AB - Psycholinguistic speech production models assume that lexical stress is stored and accessed separately during phonological encoding. We address the questions of the storage and computation of lexical stress in a case study of an Italian speaking patient with an impairment of lexical stress assignment in naming, reading, and repeating single words. The patient's stress error pattern and his performance on tasks examining lexical stress in perception suggest an impairment in the retrieval of the stress pattern of irregular words. In contrast, his assignment of stress to nonstored phonological forms suggests that the computation of stress is unimpaired. PMID- 12081426 TI - Activation of multiple phoneme associates of graphemes in visual word recognition. AB - One general issue in the domain of visual word recognition is to delineate the nature of readers' knowledge of the print-sound mapping. A more specific question is to determine whether multiple grapheme-phoneme associations are available and activated during the phonological transcoding of a letter string. Evidence for the activation of irregular associations during print-to-sound transcoding, independently from lexical influences, was assessed in a letter detection task by examining performance on target-absent pseudowords. We contrasted two types of pseudowords that could be considered homophone with a real word by application of either grapheme-phoneme correspondence rules or of multiple phonemic activation. Performance on both types of homophones was compared to nonhomophone control pseudowords, strictly equivalent in terms of orthographic similarity to the base words. The finding of a homophony disadvantage for the homophones by multiple activation was interpreted as evidence for multiple phonemic activation in the print-to-sound conversion system. PMID- 12081428 TI - The cues that children use in acquiring adjectival phrases and compound nouns: evidence from bilingual children. AB - This study explores the cues used in acquisition of two semantically similar structures that are ordered differently in French and English: adjectival phrases and compound nouns. One possible prediction is that children attend primarily to meaning, so they should order both structures similarly. Another is that children attend primarily to structure, so they should learn these structures independently. Two kinds of data were obtained from eight 4-year old bilinguals: (1) spontaneous production and (2) experimental elicitation. The children were nearly perfect in their ordering of adjectival phrases, but less so in compounds. These results conform to neither prediction, leaving open the question of which cues children use in acquisition. PMID- 12081427 TI - Why is "Red Cross" different from "Yellow Cross"?: a neuropsychological study of noun-adjective agreement within Italian compounds. AB - This study investigates the performance of two Italian nonfluent aphasic patients on noun-adjective agreement in compounds and in noun phrases. A completion, a reading, and a repetition task were administered. Results show that both patients were able to correctly inflect adjectives within compounds, but not in noun phrases. Moreover, they were sensitive to constituent order (noun-adjective vs adjective-noun) within noun phrases, but less so within compounds. These results suggest differential processing for compounds as compared to noun phrases: While the latter require standard morphosyntactic operations that are often impaired in aphasic patients, the former can be accessed as whole words at the lexical level. PMID- 12081429 TI - Interaction of morphological structure and prefix transparency in the processing of Bulgarian aspectual verb forms. AB - This study investigates the processing of polymorphemic words in a highly inflected language, Bulgarian. The roles of semantic transparency, aspect, and bound or free root status in the recognition of aspectual verb forms were probed in the visual modality in a simple lexical decision and a masked priming experiment at a short SOA. Results from the two experiments yielded effects of semantic transparency and morphological complexity, demonstrating that both factors influence the recognition of prefixed aspectual forms in Bulgarian and pointing toward different access procedures. In contrast, the status of the root did not influence recognition patterns, suggesting that free-standing and bound roots are equivalent lexical units of access and representation in Bulgarian. PMID- 12081430 TI - Neuromagnetic evidence for the timing of lexical activation: an MEG component sensitive to phonotactic probability but not to neighborhood density. AB - Evidence from electrophysiological measures such as ERPs (event-related potentials) and MEG (magnetoencephalography) suggest that the first evoked brain response component sensitive to stimulus properties affecting reaction times in word recognition tasks occurs at 300-400 ms. The present study used the stimulus manipulation of Vitevich and Luce (1999) to investigate whether the M350, an MEG response component peaking at 300-400 ms, reflects lexical or postlexical processing. Stimuli were simultaneously varied in phonotactic probability, which facilitates lexical activation, and in phonological neighborhood density, which inhibits the lexical decision process. The present results indicate that the M350 shows facilitation by phonotactic probability rather than inhibition by neighborhood density. Thus the M350 cannot be a postlexical component. PMID- 12081431 TI - Exploring word recognition in a semi-alphabetic script: the case of Devanagari. AB - Unlike other writing systems that are readily classifiable as alphabetic or syllabic in their structure, the Indic Devanagari script (of which Hindi is an example) has properties of both syllabic and alphabetic writing systems. Whereas Devanagari consonants are written in a linear left-to-right order, vowel signs are positioned nonlinearly above, below, or to either side of the consonants. This fact results in certain words in Hindi for which, in a given syllable, the vowel precedes the consonant in writing but follows it in speech. The current research exploited this property of the script to examine when the disparity between spatial and temporal sequencing would incur a processing cost and the implications of the findings from naming speed, accuracy, and writing order for the level at which words in Devanagari are segmented. The results support a partly phonemic and partly syllabic level of segmentation, consistent with the structural hybridity of the script. PMID- 12081432 TI - The bilingual mental lexicon and speech production process. AB - The Chinese/English intrasentential code-switching data provide evidence that the bilingual mental lexicon involves language contact between language-specific semantic/pragmatic feature bundles. Lemmas in the mental lexicon are tagged for specific languages and contain semantic, syntactic, and morphological information about lexemes. In a bilingual mode, the speaker makes choices at the preverbal level of lexical-conceptual structure, and these choices activate the lemmas in the mental lexicon for the speaker's preverbal message to be morphosyntactically realized at the functional level of predicate-argument structure. The result will be language-specific surface forms at the positional level of morphological realization patterns. The languages involved in the bilingual's mixed speech are never equally activated, with one language projecting the sentential frame and the other supplying a particular type of morphemes for the speaker's communicative intentions. PMID- 12081433 TI - Linking elements in Dutch noun-noun compounds: constituent families as analogical predictors for response latencies. AB - This study addresses the choice of linking elements in novel Dutch noun-noun compounds. Previous off-line experiments (Krott, Baayen, & Schreuder, 2001) revealed that this choice can be predicted analogically on the basis of the distribution of linking elements in the left and right constituent families, i.e., the set of existing compounds that share the left (or right) constituent with the target compound. The present study replicates the observed graded analogical effects under time pressure, using an on-line decision task. Furthermore, the analogical support of the left constituent family predicts response latencies. We present an implemented interactive activation network model that accounts for the experimental data. PMID- 12081434 TI - Lexical and relational influences on the processing of novel compounds. AB - To interpret a novel compound (e.g., chocolate twig), one must access the concepts denoted by the words and select a relation that links them together. To examine the role of lexical and relation information on conceptual combination, target combinations were preceded by one of three prime combinations. In Experiment 1, the prime used a semantically similar head noun and either the same or different relation. The third prime was semantically unrelated to the target. Experiment 2 was identical, except the modifier was the semantically related constituent. Although semantic priming was observed in both experiments, relation priming was obtained only when the modifier was similar. PMID- 12081435 TI - The role of typological variation in the processing of interfixed compounds. AB - This study investigates the processing of interfixed compounds within a primed lexical decision paradigm. Results from an experiment on German are compared with those from a previous study on Greek and Polish, demonstrating that patterns of morphological priming in German differ from those obtained in the other two languages. In all experiments compounds were primed by initial morphemes with and without the interfix. In Greek and Polish priming was significantly facilitated in cases where the mophological prime was homophonous with a real word. In German, however, the effect of wordness was found for un-interfixed but not for interfixed primes. Our data suggest that patterns of morphological priming for a given structure may not be generalizable to analogous structures in other languages without a consideration of language-specific morphological properties. PMID- 12081437 TI - Short planar gradient coils for MR microscopy using concentric return paths. AB - The aim of this work is to design a set of gradient coils with an optimal geometry for magnetic resonance microscopy studies. Designs for a three-axis gradient coil system particularly suited for studies with small radiofrequency coils are presented. The novel geometry involves a planar section with concentric return paths to keep the coil short. Reduction of the external field has been attempted by varying the positions of the return paths using a simulated annealing algorithm. A biplanar version of x- and z-directed prototype gradients was built and tested. A 2D-MR image of a grid phantom has been obtained on a 7-T MR instrument to demonstrate the theory. A three-axis set used as a surface gradient set has also been built and used to obtain high-resolution MR images. PMID- 12081438 TI - Correlation of backbone amide and side-chain (13)C resonances in perdeuterated proteins. AB - Side-chain carbon resonance assignments are difficult to obtain for larger proteins. While standard methods require protons for excitation and detection of magnetization, their presence is often unacceptable and often leads to unacceptable relaxation losses at the directly bound carbon sites. In this paper, pulse sequences are presented which provide connectivities between aliphatic side chain (13)C and amide (1)H and (15)N chemical shifts in fully deuterated, (13)C/(15)N-enriched proteins. Magnetization either starts off from carbons or from both nitrogens and protons and is passed along the side-chain via (13)C (13)C isotropic mixing. Direct rather than (13)CO-relayed (15)N-->(13)C(alpha) or (13)C(alpha)-->(15)N transfer steps allow the detection of intraresidual as well as sequential correlations. To avoid ambiguities between these two types in the three-dimensional version of the experiments, a fourth dimension can be introduced to achieve their separation along a (13)C(alpha) frequency axis. The novel methods are demonstrated with the uniformly (2)H/(13)C/(15)N labeled 35-kDa protein diisopropylfluorophosphatase from Loligo vulgaris. PMID- 12081439 TI - WAVEWAT-improved solvent suppression in NMR spectra employing wavelet transforms. AB - WAVEWAT is a new processing algorithm to suppress the on-resonance water signal in NMR spectra. It is based on a multiresolution analysis (MRA) of the free induction decay (FID) using a dyadic discrete wavelet transform (DWT). The width of the suppressed signal can be adjusted so that signals close to water are recovered without distortion of the signal shape and intensity. Computational efficiency is comparable to that of convolution filters employing a Fourier transform. PMID- 12081440 TI - Universal equations for linear adiabatic pulses and characterization of partial adiabaticity. AB - A numerical analysis of the sech/tanh (or hyperbolic secant) and tanh/tan adiabatic inversion pulses provides a set of master equations for each type of pulse that guarantee their optimal implementation over a wide range of practical conditions without needing to further simulate the inversion profiles of the pulses. These simple equations determine the necessary maximum RF amplitude (RF(max)) required for a preselected degree of inversion across a chosen effective bandwidth (bw(eff)) and for a chosen pulse length (T(p)). The two types of pulse function differently: The sech/tanh pulse provides a rectangular inversion profile with bw(eff) being a large fraction of the adiabatic frequency sweep (bwdth), whereas for tanh/tan bw(eff) is < or =bwdth/20. If the quality of inversion is defined as the minimum allowable extent of inversion, iota(bw), at the boundaries of bw(eff), two basic linear equations are found for both types of pulse and these are of the form (RF(max)T(p))(2)=m(1)T(p)bwdth+c(1) and T(p)bwdth=m(3)T(p)bw(eff)+c(3). The different behavior of the two pulses is expressed as different dependencies of the slopes m(n) and intercepts c(n) on iota(bw) and allowances are made for second order effects within these equations. The availability of these master relationships enables a direct comparison of the two types of adiabatic pulse and it is found that tanh/tan requires about half the pulse length of an equivalent sech/tanh pulse and also has the advantage of being less sensitive to the effects of scalar coupling. In contrast sech/tanh delivers about half the total RF power of an equivalent tanh/tan pulse. It is expected that the forms of these two basic linear equations are generally applicable to adiabatic inversion pulses and thus define the concept of "linear adiabaticity." At low values of T(p)bwdth or T(p)bw(eff) the linear equations no longer apply, defining a region of "partial adiabaticity." Normal adiabatic pulses in the middle of this partial region are more efficient in terms of RF(max) or T(p) but are moderately less tolerant to RF inhomogeneity. A class of numerically optimized pulses has recently been developed that specifically trades adiabaticity in an attempt to gain RF(max) or T(p) efficiency. In comparison to normal adiabatic pulses implemented under optimal conditions, these new partially adiabatic pulses show only marginal improvements; they are restricted to single values of T(p)bw(eff), and they are vastly less tolerant to RF inhomogeneity. These comparisons, and direct comparisons between any types of inversion pulse, adiabatic or otherwise, can be made using plots of (RF(max)T(p))(2) or (Total Power) T(p) versus T(p)bw(eff). PMID- 12081441 TI - Enhanced signal intensities obtained by out-of-phase rapid-passage EPR for samples with long electron spin relaxation times. AB - To understand the signals that are observed under rapid-passage conditions for samples with long electron spin relaxation times, the E' defect in irradiated vitreous SiO(2) was studied. For these samples at room temperature, T(1) is 200 mciro s and T(2) ranged from 35 to 200 micro s, depending on spin concentration. At X band with 100-kHz modulation frequency and 1-G modulation amplitude there was minimal lineshape difference between the low-power, in-phase spectra and high power spectra detected 90 degrees out-of-phase with respect to the magnetic field modulation. Signal enhancement, defined as the ratio of the intensities of the out-of-phase to the in-phase signals when B(1) for both observation modes is adjusted to give maximum signal, was 3.4 to 9.5 at room temperature. The origin of the out-of-phase signal was modeled by numerical integration of the Bloch equations including magnetic field modulation. The waveforms for the E' signal, prior to phase sensitive detection, were simulated by summing the contributions of many individual spin packets. Good agreement was obtained between experimental and calculated waveforms. At low B(1) the experimental values of T(1) and T(2) were used in the simulations. However, at higher B(1), T(2) was adjusted to match the experimental signal intensity and increased with increasing B(1). At high B(1), T(2)=T(1), consistent with Redfield's and Hyde's models. For the spin concentrations examined, the out-of-phase signals at very high power (B(1) approximately 0.33 G) displayed a linear relationship between peak-to-peak signal amplitude and spin concentration. Under the conditions used for spin quantitation the signal-to-noise for these spectra was up to 5 times higher than for the in phase signal, which greatly facilitates quantitation for these types of samples. For samples in which T(2) is dominated by electron spin-spin interaction, lower spin concentration results in longer T(2) and the enhancement is increased. PMID- 12081442 TI - Deconvolution of compartmental water diffusion coefficients in yeast-cell suspensions using combined T(1) and diffusion measurements. AB - An NMR method is presented for measuring compartment-specific water diffusion coefficient (D) values. It uses relaxography, employing an extracellular contrast reagent (CR) to distinguish intracellular (IC) and extracellular (EC) (1)H(2)O signals by differences in their respective longitudinal (T(1)) relaxation times. A diffusion-weighted inversion-recovery spin-echo (DW-IRSE) pulse sequence was used to acquire IR data sets with systematically and independently varying inversion time (TI) and diffusion-attenuation gradient amplitude (g) values. Implementation of the DW-IRSE technique was demonstrated and validated using yeast cells suspended in 3 mM Gd-DTPA(2-) with a wet/dry mass ratio of 3.25:1.0. Two-dimensional (2D) NMR data were acquired at 2.0 T and analyzed using numerical inverse Laplace transformation (2D- and sequential 1D-ILT) and sequential exponential fitting to yield T(1) and water D values. All three methods gave substantial agreement. Exponential fitting, deemed the most accurate and time efficient, yielded T(1):D (relative contribution) values of 304 ms:0.023x10(-5) cm(2)/s (47%) and 65 ms:1.24x10(-5) cm(2)/s (53%) for the IC and EC components, respectively. The compartment-specific D values derived from direct biexponential fitting of diffusion-attenuation data were also in good agreement. Extension of the DW-IRSE method to in vivo models should provide valuable insights into compartment-specific water D changes in response to injury or disease. (c) 2002 Elsevier Science (USA). PMID- 12081443 TI - Multiple-quantum NMR spectra of partially oriented indene: a new approach to estimating order in a nematic phase. AB - Multiple-quantum proton NMR spectra of the 8-spin system of indene partially oriented in a nematic mesophase were acquired using a magnetic field gradient (MFG) method. Six-, seven- and eight-quantum spectra were solved to yield all intramolecular dipolar couplings (D(ij)) which were used to determine the molecular shape. The spectral widths of multiple-quantum spectra are dependent on molecular orientation and geometry and a novel method was developed to estimate the Saupe order parameters from the observed spectral widths of the high-order multiple-quantum spectra. PMID- 12081444 TI - Multiple spin echoes in heterogeneous systems: physical origins of the observed dips. AB - Dipolar interactions in liquids have recently offered a new challenge to investigate porous media by exploiting intermolecular quantum coherences, which are obtained through a simple two-pulse sequence (90 degrees -tau-120 degrees ). This sequence, in the presence of an external gradient (G), refocuses a train of echoes at multiple integer values of time tau. The first and second echo amplitudes are acquired for heterogeneous systems such as porous media at different time values (tau). In our first experiments on bovine bone samples we have observed unpredicted dips on the second echo time behavior. We argue that a strict relation occurs between the average pore dimensions and the dips time position through the correlation distance d=pi/(gamma G tau) (defined as half a cycle of the magnetization helix, which originates in the presence of an external gradient). Although the experimental results have revealed an exceptional connection between the porous structure and the correlation distance, no physical explanation was so far provided. In this paper we propose a possible physical cause of the observed phenomenon. In addition we report an accurate analysis of new experiments performed on glass beads phantoms, which confirms our conclusions. PMID- 12081445 TI - Homonuclear zero-quantum recoupling in fast magic-angle spinning nuclear magnetic resonance. AB - Solid-state magic-angle-spinning NMR pulse sequences which implement zero-quantum homonuclear dipolar recoupling are designed with the assistance of symmetry theory. The pulse sequences are compensated on a short time scale by the use of composite pulses and on a longer time scale by the use of supercycles. (13)C dipolar recoupling is demonstrated in powdered organic solids at high spinning frequencies. The new sequences are compared to existing pulse sequences by means of numerical simulations. Experimental two-dimensional magnetization exchange spectra are shown for [U-(13)C]-L-tyrosine. PMID- 12081446 TI - NMR difference probe: a dual-coil probe for NMR difference spectroscopy. AB - A unique probe designed to acquire nuclear magnetic resonance difference spectra of two samples is presented. The NMR Difference Probe contains two sample coils in a resonant circuit that switches between parallel excitation and serial acquisition to cancel common signals such as solvent peaks and impurities. Two samples containing a common analyte, acetonitrile, were used to demonstrate signal cancellation in a difference spectrum collected with a single pulse experiment. The cancellation was over 96% effective. The approach described has applications in the areas of solvent subtraction and spectral simplification. PMID- 12081447 TI - Structural determination of spin label immobilization and orientation: a Monte Carlo minimization approach. AB - Electron paramagnetic resonance (EPR) is often used in the study of the orientation and dynamics of proteins. However, there are two major obstacles in the interpretation of EPR signals: (a) most spin labels are not fully immobilized by the protein, hence it is difficult to distinguish the mobility of the label with respect to the protein from the reorientation of the protein itself; (b) even in cases where the label is fully immobilized its orientation with respect to the protein is not known, which prevents interpretation of probe reorientation in terms of protein reorientation. We have developed a computational strategy for determining whether or not a spin label is immobilized and, if immobilized, predicting its conformation within the protein. The method uses a Monte Carlo minimization algorithm to search the conformational space of labels within known atomic level structures of proteins. To validate the method a series of spin labels of varying size and geometry were docked to sites on the myosin head catalytic and regulatory domains. The predicted immobilization and conformation compared well with the experimentally determined mobility and orientation of the label. Thus, probes can now be targeted to report on various modes of molecular dynamics: immobilized probes to report on protein backbone and domain dynamics or floppy probes to report on the extent of steric restriction experienced by the side chain. PMID- 12081448 TI - Frequency dependence of EPR signal intensity, 250 MHz to 9.1 GHz. AB - Experimental EPR signal intensities at 250 MHz, 1.5 GHz, and 9.1 GHz agree within experimental error with predictions from first principles. When both the resonator size and the sample size are scaled with the inverse of RF/microwave frequency, omega, the EPR signal at constant B(1) scales as omega(-1/4). Comparisons were made for three different samples in two pairs of loop gap resonators. Each pair was geometrically scaled by a factor of 6. One pair of resonators was scaled from 250 MHz to 1.5 GHz, and the other pair was scaled from 1.5 GHz to 9 GHz. All terms in the comparison were measured directly, and their uncertainties estimated. The theory predicts that the signal at the lower frequency will be larger than the signal at the higher frequency by the ratio 1.57. For 250 MHz to 1.5 GHz, the experimental ratio was 1.52 and for the 1.5-GHz to 9-GHz comparison the ratio was 1.14. PMID- 12081449 TI - Water content profiles with a 1D centric SPRITE acquisition. AB - The purpose of this work is to develop a rapid MRI method amenable to profiling with minimal or no T(1) relaxation weighting. The behavior of a signal during a centric SPRITE acquisition is analyzed. It is shown that the technique can be made immune to a broad range of T(1) changes. In a properly executed measurement, only T(2)* and proton density parameters define the image intensity. A T(2)* mapping technique can be easily applied, separating T(2)* and proton density contributions to the image. A drying soil sample with low initial water content is experimentally studied as a demonstration of the technique. A characteristic baseline artifact is easily removed from the profiles by a simple operation. PMID- 12081450 TI - Optimizing STMAS. AB - The 2D satellite transition magic angle spinning (STMAS) experiment generates efficiently high-resolution isotropic NMR spectra of half-integer quadrupolar nuclei. The experiment involves excitation and coherence transfer of satellite transitions into the central transition. It requires efficient refocusing of satellite transitions and sample spinning at a very accurate magic angle to cancel the first-order quadrupolar interaction effect. A review of all parameters relevant to optimizing the STMAS experiment is presented, including pulse sequence calibration, regulating spinning speed, magic angle adjustment, optimization of satellite transition excitation, and coherence transfer for both I = 3/2 and I > or =5/2 nuclei. PMID- 12081451 TI - The 2D-J-DOSY experiment: resolving diffusion coefficients in mixtures. AB - Many diffusion-ordered spectroscopy (DOSY) NMR techniques have recently been developed to aid in the deconvolution of complex mixtures. Spectroscopic separation based on chemical and physical properties facilitates the identification of mixture components while eliminating time-consuming separation steps and preserving the chemical environment. One way to improve resolution in such experiments is to spread the spectroscopic information into two dimensions. The 2D-J-DOSY experiment has been designed to resolve mixture components in terms of a chemical shift and proton coupling constant as well as distinguishing them on the basis of translational diffusion. Acquiring a series of spectra as a function of gradient amplitude permits the determination of diffusion coefficients for components that cannot be resolved in the one-dimensional (1D) (1)H NMR spectrum. Comparison of the resulting values with those obtained through the traditional 1D diffusion experiment for a mixture of sugars validates The 2D J-DOSY technique. PMID- 12081452 TI - Two-dimensional high-resolution NMR spectra in matched B(0) and B(1) field gradients. AB - In a recent publication we presented a method to obtain highly resolved NMR spectra in the presence of an inhomogeneous B(0) field with the help of a matched RF gradient. If RF gradient pulses are combined with "ideal" 90 degrees pulses to form inhomogeneous z rotation pulses, the line broadening caused by the B(0) gradient can be refocused, while the full chemical shift information is maintained. This approach is of potential use for NMR spectroscopy in an inhomogeneous magnetic field produced by an "ex-situ" surface spectrometer. In this contribution, we extend this method toward two-dimensional spectroscopy with high resolution in one or both dimensions. Line narrowing in the indirect dimension can be achieved by two types of nutation echoes, thus leading to depth sensitive NMR spectra with full chemical shift information. If the nutation echo in the indirect dimension is combined with a stroboscopic acquisition using inhomogeneous z-rotation pulses, highly resolved two-dimensional correlation spectra can be obtained in matched field gradients. Finally, we demonstrate that an INEPT coherence transfer from proton to carbon spins is possible in inhomogeneous B(0) fields. Thus, it is possible to obtain one-dimensional (13)C NMR spectra with increased sensitivity and two-dimensional HETCOR spectra in the presence of B(0) gradients of 0.4 mT/cm. These schemes may be of some value for ex-situ NMR analysis of materials and biological systems. PMID- 12081453 TI - A 4-mm probe for (13)C CP/MAS NMR of solids at 21.15 T. AB - The design of a broadband 4-mm magic-angle spinning (MAS) X-(1)H/(19)F double resonance probe for cross-polarization (CP)/MAS NMR studies at 21.15 T ((1)H at 900 MHz) is described. The high-frequency (1)H/(19)F channel employs a new and efficient transmission line tuning design. The first (13)C CP/MAS NMR spectra recorded at 21.15 T have been obtained with this probe and exhibit the best S/N per milligram sample of hexamethylbenzene achieved so far for a 4-mm rotor. PMID- 12081454 TI - Intraresidue HNCA and COHNCA experiments for protein backbone resonance assignment. AB - Two novel experiments, intra-HNCA and intra-COHNCA, are presented for sequential backbone resonance assignment of (13)C, (15)N labeled proteins. The advantage with respect to conventional pulse schemes is the suppression of the sequential (15)N-->(13)C(alpha) coherence transfer pathway, which can be separately obtained from a HNCOCA correlation experiment. This results in a two-fold reduction of the number of detected correlation peaks. Spectral simplification is especially important for efficient automated assignment protocols as required in the context of high-throughput protein studies by NMR. The performance of the new experiments is demonstrated on an 18-kDa protein fragment of the E. coli sulfite reductase and compared to conventional techniques in terms of sensitivity and resolution. PMID- 12081455 TI - Phoneme awareness is a better predictor of early reading skill than onset-rime awareness. AB - We present the results of a short-term longitudinal study. Children in the early stages of learning to read (5 and 6 year olds) were administered three different tasks (deletion, oddity, and detection) tapping awareness of four phonological units (initial phoneme, final phoneme, onset, and rime). Measures of phoneme awareness were the best concurrent and longitudinal predictors of reading skill with onset-rime skills making no additional predictive contribution once phonemic skills were accounted for. The findings are related to recent controversy over the role of large versus small phonological units as predictors of children's reading skills. PMID- 12081456 TI - Reflections on onset-rime and phoneme sensitivity as predictors of beginning word reading. AB - Rival claims have been made concerning the importance of rime sensitivity as a predictor of early word reading skill. Hulme et al. (2002) suggested that phoneme sensitivity is more strongly predictive of word reading ability than is onset rime sensitivity. An examination of two independent data sets suggests that, although onset-rime sensitivity typically predicts school entrants' later word reading skill, phoneme sensitivity does predict more variation. However, multiple regression analyses do not reveal the level of phonological sensitivity that children need in order to understand alphabetic reading instruction. This issue is crucial to the detection of children at risk for reading failure and for the design of intervention programs for these children. A different analytic strategy is described for addressing this issue. PMID- 12081457 TI - It doesn't matter whether onset and rime predicts reading better than phoneme awareness does or vice versa. AB - Hulme et al. argue against our hypothesis that there are two routes from onset and rime awareness to reading: an indirect route whereby onset-rime awareness feeds into the development of phoneme awareness which in turn affects children's reading, and a direct route by which onset-rime awareness makes an independent contribution to children's reading. The evidence that Hulme et al. present against this hypothesis is not convincing, partly because our hypothesis actually predicts most of their results and partly because of weaknesses in the design of Hulme et al.'s study and in the unusual procedures that they employed. PMID- 12081458 TI - In the beginning was the rhyme? A reflection on Hulme, Hatcher, Nation, Brown, Adams, and Stuart (2002). AB - Phonological sensitivity at different grain sizes is a good predictor of reading acquisition in all languages. However, prior to any explicit tuition in alphabetic knowledge, phonological sensitivity develops at the larger grain sizes syllables, onsets, and rimes-in all languages so far studied. There are also developmental differences in the grain size of lexical representations and reading strategies across orthographies. Phoneme-level skills develop fastest in children acquiring orthographically consistent languages with a simple syllabic (CV) structure, such as Finnish and Italian. For English, however, both "large" and "small" units are important for the successful acquisition of literacy. PMID- 12081460 TI - Structure of preschool phonological sensitivity: overlapping sensitivity to rhyme, words, syllables, and phonemes. AB - Phonological sensitivity is an important causal variable in reading acquisition; however, there is controversy concerning its nature. One view holds that sensitivity to various linguistic units reflects independent abilities, whereas another holds sensitivity to these units reflects one ability. We examined relations among sensitivity to words, syllables, rhymes, and phonemes in 149 older preschool children (4- and 5-year-olds) and 109 younger preschool children (2- and 3-year-olds) who completed eight measures of phonological sensitivity and measures of print knowledge. Confirmatory factor analyses of all combinations of word, syllable, rhyme, and phoneme factors found that a one-factor model best explained the data from both groups of children (CFIs >.98). Only variance common to all phonological sensitivity skills was related to print knowledge and rudimentary decoding. Findings support a developmental conceptualization of phonological sensitivity. PMID- 12081461 TI - Protein import into mitochondria of Neurospora crassa. AB - Biogenesis of mitochondria requires import of several hundreds of different nuclear-encoded preproteins needed for mitochondrial structure and function. Import and sorting of these preproteins is a multistep process facilitated by complex proteinaceous machineries located in the mitochondrial outer and inner membranes. The translocase of the mitochondrial outer membrane, the TOM complex, comprises receptors which specifically recognize mitochondrial preproteins and a protein conducting channel formed by TOM40. The TOM complex is able to insert resident proteins into the outer membrane and to translocate proteins into the intermembrane space. For import of inner membrane or matrix proteins, the TOM complex cooperates with translocases of the inner membrane, the TIM complexes. During the past 30 years, intense research on fungi enabled the identification and mechanistic characterization of a number of different proteins involved in protein translocation. This review focuses on the contributions of the filamentous fungus Neurospora crassa to our current understanding of mitochondrial protein import, with special emphasis on the structure and function of the TOM complex. PMID- 12081462 TI - Mitochondrial dynamics in filamentous fungi. AB - Mitochondria are essential organelles of eukaryotic cells. They grow continuously throughout the cell cycle and are inherited by daughter cells upon cell division. Inheritance of mitochondria and maintenance of mitochondrial distribution and morphology require active transport of the organelles along the cytoskeleton and depend on membrane fission and fusion events. Many of the molecular components and cellular mechanisms mediating these complex processes have been conserved during evolution across the borders of the fungal and animal kingdoms. During the past few decades, several constituents of the cellular machinery mediating mitochondrial behavior have been identified and functionally characterized. Here, we review the contributions of fungi, with special emphasis on the filamentous fungus Neurospora crassa, to our current understanding of mitochondrial morphogenesis and inheritance. PMID- 12081463 TI - Isogamous, hermaphroditic inheritance of mitochondrion-encoded resistance to Qo inhibitor fungicides in Blumeria graminis f. sp. tritici. AB - A mutation of glycine to alanine at position 143 in the mitochondrial cytochrome b amino acid sequence of Blumeria graminis f. sp. tritici cosegregated with the QoI-resistant phenotype in a ratio of 1:1 in a cross between a sensitive and a resistant isolate. This mutation was used as a mitochondrial marker to determine whether mitochondrial inheritance in B. graminis was anisogamous, as in heterothallic Neurospora sp., or isogamous and hermaphroditic, as in Aspergillus nidulans. Segregation of mitochondrial genotypes in B. graminis f. sp. tritici was consistent with inheritance of mitochondria being hermaphroditic and isogamous, in that all ascospores from an individual cleistothecium had the same mitochondrial genotype and that either parent could act as the maternal parent of a cleistothecium. Within each cleistothecium, nuclear segregation occurred independently of mitochondrial inheritance, as shown by segregation of resistance to the fungicide triadimenol and by segregation of avirulences to the wheat cultivars Galahad (Pm2), Armada (Pm4b), and Holger (Pm6). PMID- 12081464 TI - Functional analysis of the C6 zinc finger gene pro1 involved in fungal sexual development. AB - The pro1 gene, controlling fruiting body development in the homothallic ascomycete Sordaria macrospora, encodes a C6 zinc finger protein with a typical DNA binding domain of GAL4-like C6 zinc finger proteins as well as a putative nuclear targeting signal. In the corresponding mutant pro1, the pro1 gene is deleted, and the transition of primordia into mature fruiting bodies is prevented. To further characterize the PRO1 polypeptide, the yeast system was used for identifying a transactivation domain in the N-terminal half of PRO1, which probably also functions in S. macrospora. The functional analysis was extended by using truncated versions of the pro1 gene in complementation transformations of a deltapro1 mutant. Interestingly, the 5' part of the pro1 gene encoding the DNA binding and transactivation domain as well as putative nuclear targeting signals was sufficient to restore fertility in the sterile pro1 mutant. In vitro mutagenesis verified that the DNA binding domain is essential for normal fruiting body development. This was concluded from transformation experiments with eight pro1 derivatives containing triplet substitutions in conserved codons of the DNA binding domain; some, but not all, failed in restoring the wild-type phenotype in mutant pro1. Using a PCR-based cloning strategy, pro1 homologs from the two related heterothallic species Neurospora crassa and Sordaria brevicollis were isolated, showing similarities in the predicted amino acid sequences of 91 and 90%, respectively. When a N. crassa pro1 cDNA clone was used in complementation transformations, we succeeded in restoring the wild-type phenotype to the S. macrospora pro1 mutant. These data suggest that pro1 homologs from heterothallic species can provide the pro1 function in homothallic ascomycetes. Based on the published sequence of the N. crassa genome, we identified hpro1A, another transcriptionally expressed gene, with a similarity of 40% to the pro1 genes, which is present as a single copy gene in N. crassa as well as in S. macrospora. PMID- 12081465 TI - Involvement of Candida albicans NADH dehydrogenase complex I in filamentation. AB - The gene encoding the 51-kDa subunit of nicotinamide adenine dinucleotide (NADH) dehydrogenase complex I, a principal component of the mitochondrial electron transport chain, was cloned in Candida tropicalis. The homolog in C. albicans, CaNDH51, was identified, and each allele was successively disrupted by PCR mediated gene disruption. Wild type, heterozygote, reintegrant, and homozygous null mutants grew as blastoconidia in rich medium containing 3% glucose, but the homozygous null mutant failed to grow in ethanol or acetate. When glucose concentration was varied from 1 mM (0.018%) to 200 mM (3.6%) in a basal salts medium, all strains grew equally well at all glucose concentrations; the wild type strain, the heterozygote, and the reintegrant exhibited abundant germ tubes, pseudohyphae, and hyphae. In contrast, the ndh51/ndh51 strain failed to display any type of filamentous growth, even in glucose concentrations as low as 1 mM. These results suggest a previously unexplored relationship between mitochondrial electron transport and morphogenesis. PMID- 12081466 TI - Distribution of mating type alleles in the wheat pathogen Mycosphaerella graminicola over spatial scales from lesions to continents. AB - A total of 2035 Mycosphaerella graminicola strains collected from 16 geographic locations on four continents were assayed for the mating type locus. RFLP fingerprints were used to identify clones in each population. At the smallest spatial scale analyzed, both mating types were found among fungal strains sampled from different lesions of the same leaf as well as from different pycnidia in the same lesion. At larger spatial scales, the two mating types were found at equal frequencies across spatial scales ranging from several square meters to several thousand square kilometers. Though the absolute frequencies of the two mating types sometimes varied for different sampling units within the same spatial scale in the hierarchy (plots within a field, fields within a country, or different continents of the world), none of the differences were statistically significant from the null hypothesis of equal frequencies for the two mating types. The evolutionary forces likely to maintain the even distribution of the two mating types in this pathogen were discussed. PMID- 12081467 TI - Differential expression of G protein alpha and beta subunit genes during development of Phytophthora infestans. AB - A G protein alpha subunit gene (pigpa1) and a G protein beta subunit gene (pigpb1) were isolated from the oomycete Phytophthora infestans, the causal agent of potato late blight. Heterotrimeric G proteins are evolutionary conserved GTP binding proteins that are composed of alpha,beta, and gamma subunits and participate in diverse signal transduction pathways. The deduced amino acid sequence of both pigpa1 and pigpb1, showed the typical conserved motifs present in Galpha or Gbeta proteins from other eukaryotes. Southern blot analysis revealed no additional copies of Galpha or Gbeta subunit genes in P. infestans, suggesting that pigpa1 and pigpb1 are single copy genes. By cross-hybridization homologues of gpa1 and gpb1 were detected in other Phythophthora species. Expression analyses revealed that both genes are differentially expressed during asexual development, with the highest mRNA levels in sporangia. In mycelium, no pigpa1 mRNA was detected. Western blot analysis using a polyclonal GPA1 antibody confirmed the differential expression of pigpa1. These expression patterns suggest a role for G-protein-mediated signaling during formation and germination of asexual spores of P. infestans, developmental stages representing the initial steps of the infection process. PMID- 12081468 TI - Transmission of double-stranded RNA in Heterobasidion annosum. AB - Transmission of dsRNA viruses between homo- and heterokaryotic mycelia paired on agar plates and into conidia has been studied in Heterobasidion annosum. Horizontal transmission of dsRNA occurred between both homo- and heterokaryotic isolates, as well as between isolates belonging to different intersterility groups. The proportions of vertical transmission into conidia were 3% and 55%, respectively, for the two isolates included in the study. RT-PCR of dsRNA and PCR RFLP of mitochondrial markers were used to confirm transmission of dsRNA between the cytoplasms of different mycelia. The identity of nuclei and nuclear migration during experiments were verified using PCR-RFLP of several nuclear markers. PMID- 12081469 TI - The SOD2 gene, encoding a manganese-type superoxide dismutase, is up-regulated during conidiogenesis in the plant-pathogenic fungus Colletotrichum graminicola. AB - The SOD2 gene, encoding a manganese-type superoxide dismutase (MnSOD), was identified from Colletotrichum graminicola among a collection of cDNAs representing genes that are up-regulated during conidiogenesis. The SOD2 gene consists of a 797-bp open reading frame that is interrupted by three introns and is predicted to encode a polypeptide of 208 amino acids. All conserved residues of the MnSOD protein family, including four consensus metal binding domains, are present in the predicted SOD2 protein. However, the predicted protein does not appear to contain a signal peptide that would target it to the mitochondria. Northern hybridizations revealed that expression of the approximately 900-bp SOD2 transcript is closely associated with differentiation of both oval and falcate conidia. Southern analysis indicated that there is only a single copy of the gene. SOD2 disruption strains were morphologically and pathogenically indistinguishable from wild-type strains. The dispensability of the MnSOD enzyme may be due to the activities of two other SOD enzymes, a highly expressed iron type superoxide dismutase and a much less abundant copper/zinc type, that were also detected in C. graminicola. PMID- 12081470 TI - Multiple conformational changes in enzyme catalysis. AB - Understanding the molecular mechanisms of enzyme catalysis and allosteric regulation has been a primary goal of biochemistry for many years. The dynamics of these processes, approached through a variety of kinetic methods, are discussed. The results obtained for many different enzymes suggest that multiple intermediates and conformations are general characteristics of the catalytic process and allosteric regulation. Ribonuclease, dihydrofolate reductase, chymotrypsin, aspartate aminotransferase, and aspartate transcarbamoylase are considered as specific examples. Typical and maximum rates of conformational changes and catalysis are also discussed, based on results obtained from model systems. The nature and rates of interconversion of the intermediates, along with structural information, can be used as the bases for understanding the incredible catalytic efficiency of enzymes. Potential roles of conformational changes in the catalytic process are discussed in terms of static and environmental effects, and in terms of dynamic coupling within the enzyme-substrate complex. PMID- 12081471 TI - Structural aspects of aldehyde dehydrogenase that influence dimer-tetramer formation. AB - Aldehyde dehydrogenases are isolated as dimers or tetramers but have essentially identical structures. The homotetramer (ALDH1 or ALDH2) is a dimer of dimers (A-B + C-D). In the tetrameric enzyme, Ser500 from subunit "D" interacts with Arg84, a conserved residue, from subunit "A". In the dimeric ALDH3 form, the interaction cannot exist. It has been proposed that the formation of the tetramer is prevented by the presence of a C-terminal tail in ALDH3 that is not present in ALDH1 or 2. To understand the forces that maintain the tetramer, deletion of the tail in ALDH3, addition of different tails in ALDH1, and mutations of different residues located in the dimer-dimer interface were made. Gel filtration of the recombinantly expressed enzymes demonstrated that no change in their oligomerization occurred. Urea denaturation showed a diminution to the stability of the ALDH1 mutants. The K(m) for propionaldehyde was similar to that of the wild-type enzyme, but the K(m) for NAD was altered. A double mutant of D80G and S82A produced an enzyme with the ability to form dimers and tetramers in a protein-concentration-dependent manner. Though stable, this dimeric form was inactive. The tetramer exhibited 10% activity compared with the wild type. Sequence alignment demonstrated that the hydrophobic surface area is increased in the tetrameric enzymes. The hydrophobic surface seems to be the main force that drives the formation of tetramers. The results indicated that residues 80 and 82 are involved in maintaining the tetramer after its assembly but the C-terminal extension contributes to the overall stability of the assembled protein. PMID- 12081472 TI - Conversion of a plant oxidosqualene-cycloartenol synthase to an oxidosqualene lanosterol cyclase by random mutagenesis. AB - A random mutagenesis/in vivo selection approach was applied to generate and identify mutations that alter the product specificity of oxidosqualene cycloartenol synthase (CAS) from Arabidopsis thaliana. This work complements previous studies of triterpene cyclase enzymes and was undertaken to provide knowledge of the frequency and locations at which point mutations can alter cyclase product specificity. Random mutations were introduced by treatment with hydroxylamine or passage through a mutator strain of bacteria. Libraries of mutated plasmids carrying the cas1 gene were transformed into a cyclase-deficient strain of Saccharomyces cerevisiae (CBY57) bearing a complementing plasmid (pZS11) carrying an Erg7 gene that encodes wild-type yeast oxidosqualene lanosterol cyclase and a URA3 marker that could be counterselected by growth in media containing 5-fluoroorotic acid (5-FOA). This allowed use of a plasmid shuffle to select for cas1 mutants that could substitute for ERG7 activity. Five of approximately 73,000 transformants were observed to grow in media containing 5 FOA but lacking ergosterol. pTKP5-derived plasmids isolated from these transformants were sequenced, revealing five distinct and unique point mutations: Tyr410Cys, Ala469Val, His477Tyr, Ile481Thr, and Tyr532His. Analysis of the nonsaponifiable lipids from CBY57 cells expressing these mutants suggests that the Tyr410Cys and His477Tyr mutants produce lanosterol as the dominant product, whereas the Ala469Val, Ile481Thr, and Tyr532His mutants produce a mixture of lanosterol and achilleol A, a product of monocyclization. Sequence and structural homology modeling of CAS indicate that the observed product specificity-altering mutations occur both within (Tyr410Cys, Ile481Thr, and Tyr532His) and outside of (Ala469Val and His477Tyr) the cyclase active site. PMID- 12081473 TI - The aromatic "trapping" of the catalytic histidine is essential for efficient catalysis in acetylcholinesterase. AB - While substitution of the aromatic residues (Phe295, Phe338), located in the vicinity of the catalytic His447 in human acetylcholinesterase (HuAChE) had little effect on catalytic activity, simultaneous replacement of both residues by aliphatic amino acids resulted in a 680-fold decrease in catalytic activity. Molecular simulations suggested that the activity decline is related to conformational destabilization of His447, similar to that observed for the hexamutant HuAChE which mimics the active center of butyrylcholinesterase. On the basis of model structures of other cholinesterases (ChEs), we predicted that catalytically nonproductive mobility of His447 could be restricted by introduction of aromatic residue in a different location adjacent to this histidine (Val407). Indeed, the F295A/F338A/V407F enzyme is 170-fold more reactive than the corresponding double mutant and only 3-fold less reactive than the wild-type HuAChE. However, analogous substitution of Val407 in the hexamutant HuAChE (generating the heptamutant Y72N/Y124Q/W286A/F295L/F297V/Y337A/V407F) did not enhance catalytic activity. Reactivity of these double, triple, hexa, and hepta mutant HuAChEs was monitored toward covalent ligands such as organophosphates and the transition state analogue TMFTA, which probe, respectively, the facility of the enzymes to accommodate Michaelis complexes and to undergo the acylation process. The findings suggest that in the F295A/F338A mutant the two His447 conformational states, which are essential for the different stages of the catalytic process, seem to be destabilized. On the other hand, in the F295A/F338A/V407F mutant only the state involved in acylation is impaired. Such differential effects on the His447 conformational properties demonstrate the general role of aromatic residues in cholinesterases, and probably in other serine hydrolases, in "trapping" of the catalytic histidine and thereby in optimization of catalytic activity. PMID- 12081474 TI - Binding of protoporphyrin IX and metal derivatives to the active site of wild type mouse ferrochelatase at low porphyrin-to-protein ratios. AB - Resonance Raman (RR) spectroscopy is used to examine porphyrin substrate, product, and inhibitor interactions with the active site of murine ferrochelatase (EC 4.99.1.1), the terminal enzyme in the biosynthesis of heme. The enzyme catalyzes in vivo Fe(2+) chelation into protoporphyrin IX to give heme. The RR spectra of native ferrochelatase show that the protein, as isolated, contains varying amounts of endogenously bound high- or low-spin ferric heme, always at much less than 1 equiv. RR data on the binding of free-base protoporphyrin IX and its metalated complexes (Fe(III), Fe(II), and Ni(II)) to active wild-type protein were obtained at varying ratios of porphyrin to protein. The binding of ferric heme, a known inhibitor of the enzyme, leads to the formation of a low-spin six coordinate adduct. Ferrous heme, the enzyme's natural product, binds in the ferrous high-spin five-coordinate state. Ni(II) protoporphyrin, a metalloporphyrin that has a low tendency toward axial ligation, becomes distorted when bound to ferrochelatase. Similarly for free-base protoporphyrin, the natural substrate of ferrochelatase, the RR spectra of porphyrin-protein complexes reveal a saddling distortion of the porphyrin. These results corroborate and extend our previous findings that porphyrin distortion, a crucial step of the catalytic mechanism, occurs even in the absence of bound metal substrate. Moreover, RR data reveal the presence of an amino acid residue in the active site of ferrochelatase which is capable of specific axial ligation to metals. PMID- 12081475 TI - Determination of active concentrations and association and dissociation rate constants of interacting biomolecules: an analytical solution to the theory for kinetic and mass transport limitations in biosensor technology and its experimental verification. AB - Accurate determination of kinetic rate constants for interacting biomolecules requires knowledge of the active concentrations of the participating molecules. Also, in other biomedical and clinical applications, sensitive, precise and accurate methods are needed to determine the concentration of biologically active molecules, which frequently constitute only a fraction of the total molecular pool. Here we report a novel development of the approach to determining active concentrations based on surface plasmon resonance (SPR) technology. The method relies on changes in binding rates with varying flow rates under conditions of partial mass transport, and does not require standards of known concentrations, given that the molecular mass of the molecule of interest is known. We introduce an analytical solution to the differential equations describing the formation of a 1:1 bimolecular complex, taking into account both the association and dissociation reactions, under partial mass transport limitations. This solution can be used in global fitting to binding curves obtained at different flow rates. The accuracy, precision, and sensitivity of this approach were determined in experiments involving binding of tyrosine-phosphorylated recombinant proteins to anti-phosphotyrosine antibodies, where the active concentration could be determined independently by in vitro phosphorylation with (33)P. There was an excellent agreement between the active concentrations determined by the analytical SPR-based method and by determination of the level of radioactivity of the phosphorylated protein. The SPR-based method allows determination of protein concentrations at picomolar levels. A procedure for accurate determinations of association and dissociation rate constants, based on the analytical solution of the mass transport and binding theory, is outlined. PMID- 12081476 TI - Dominant effect of protein charge rather than protein shape in apparent DNA bending by engineered bZIP domains. AB - We are interested in how asymmetric charge neutralization of DNA by proteins results in DNA bending. We have previously reported electrophoretic phasing experiments utilizing homodimer peptides derived from GCN4, a yeast basic zipper (bZIP) transcription factor. Here we report the results of experiments that examine the importance of peptide sequence context in DNA bending and test the hypothesis that peptide structural asymmetry causes electrophoretic anomalies in the absence of DNA bending. We prepared two new series of bZIP peptides that differed dramatically in overall size, structure, and peptide sequence near the DNA. The magnitude of apparent DNA bending is independent of the structure of the protein. This result reduces the concern that bZIP protein structure causes electrophoretic anomalies in the absence of DNA bending. In all cases, both the magnitude and direction of the apparent DNA bend angle are strongly dependent on the local peptide charge. We attempted to validate independently our results with a minicircle competition binding assay. Binding preferences of severalfold for properly phased circular versus linear DNA templates were predicted. However, no binding preferences were observed. We propose that the minicircle binding assay may be intrinsically insensitive to DNA bending or flexibility induced by the bZIP peptides studied, and we provide a unifying explanation for the discrepancies between the cyclization and electrophoretic experiments. PMID- 12081477 TI - Engineering autoactivating forms of matrix metalloproteinase-9 and expression of the active enzyme in cultured cells and transgenic mouse brain. AB - Matrix metalloproteinases (MMPs) are hypothesized to play an important role in the pathogenesis of several central nervous system disorders. Increased levels of expression of MMP-9 (gelatinase B) and MMP-2 (gelatinase A) have been observed in Alzheimer's disease, stroke, multiple sclerosis, and amyotrophic lateral sclerosis. This suggests an aberrant regulation of MMPs that could lead to inappropriate expression of MMP activity. To allow us to evaluate the effect of increased levels of active MMP-9 in the central nervous system, mutant forms of the enzyme were designed to autocatalytically remove the pro domain, yielding active enzyme. This was accomplished by modifying residues in the cysteine switch autoinhibitor region of the propeptide. Stable cell lines and transgenic mice that express G100L and D103N autoactive forms of human MMP-9 were developed to study the role of dysregulation of MMP-9 in disease. PMID- 12081478 TI - Evidence that helix 8 of rhodopsin acts as a membrane-dependent conformational switch. AB - The crystal structure of rhodopsin revealed a cytoplasmic helical segment (H8) extending from transmembrane (TM) helix seven to a pair of vicinal palmitoylated cysteine residues. We studied the structure of model peptides corresponding to H8 under a variety of conditions using steady-state fluorescence, fluorescence anisotropy, and circular dichroism spectroscopy. We find that H8 acts as a membrane-surface recognition domain, which adopts a helical structure only in the presence of membranes or membrane mimetics. The secondary structural properties of H8 further depend on membrane lipid composition with phosphatidylserine inducing helical structure. Fluorescence quenching experiments using brominated acyl chain phospholipids and vesicle leakage assays suggest that H8 lies within the membrane interfacial region where amino acid side chains can interact with phospholipid headgroups. We conclude that H8 in rhodopsin, in addition to its role in binding the G protein transducin, acts as a membrane-dependent conformational switch domain. PMID- 12081479 TI - Probing the interaction of bovine cytochrome P450scc (CYP11A1) with adrenodoxin: evaluating site-directed mutations by molecular modeling. AB - The present study was undertaken to evaluate the role of positively charged amino acid residues proposed to reside on the proximal surface of bovine cytochrome P450 cholesterol side chain cleavage (P450scc, CYP11A1) and to determine which residues may be involved in protein-protein interactions with the electron carrier adrenodoxin (Adx). In previous studies, nine different lysine residues were identified by chemical and immunological cross-linking experiments as potentially interacting with Adx, while in the present study, two arginine residues have been identified from sequence alignments. From these 11 residues, 13 different P450scc mutants were made of which only seven were able to be expressed and characterized. Each of the seven mutants were evaluated for their ability to bind Adx, to be reduced, and for their enzymatic activity. Among these, K403Q and K405Q showed a consistent decrease in Adx binding, the ability to be reduced by Adx, and enzymatic activity, with K405Q being affected to a much greater extent. More dramatic was the complete loss of Adx binding by R426Q, while still retaining its ability to be chemically reduced and bind carbon monoxide. Independently, a homology model of P450scc was constructed and docked with the structure of Adx. Four potential sites of interaction were identified: P450scc:K403 with Adx:D76, P450scc:K405 with Adx:D72; P450scc:R426 with Adx:E73, and P450scc:K267 with Adx:E47. Thus, the biochemical and molecular modeling studies together support the hypothesis that K267, K403, K405, and R426 participate in the electrostatic interaction of P450scc with Adx. PMID- 12081481 TI - Mutating the four extracellular cysteines in the chemokine receptor CCR6 reveals their differing roles in receptor trafficking, ligand binding, and signaling. AB - CCR6 is the receptor for the chemokine MIP-3 alpha/CCL20. Almost all chemokine receptors contain cysteine residues in the N-terminal domain and in the first, second, and third extracellular loops. In this report, we have studied the importance of all cysteine residues in the CCR6 sequence using site-directed mutagenesis and biochemical techniques. Like all G protein-coupled receptors, mutating disulfide bond-forming cysteines in the first (Cys118) and second (Cys197) extracellular loops in CCR6 led to complete elimination of receptor activity, which for CCR6 was also associated with the accumulation of the receptor intracellularly. Although two additional cysteines in the N-terminal region and the third extracellular loop, which are present in almost all chemokine receptors, are presumed to form a disulfide bond, this has not been demonstrated experimentally for any of these receptors. We found that mutating the cysteines in the N-terminal domain (Cys36) and the third extracellular loop (Cys288) neither significantly affected receptor surface expression nor completely abolished receptor function. Importantly, contrary to several previous reports, we demonstrated directly that instead of forming a disulfide bond, the N terminal cysteine (Cys36) and the third extracellular loop cysteine (Cys288) contain free SH groups. The cysteine residues (Cys36 and Cys288), rather than forming a disulfide bond, may be important per se. We propose that CCR6 forms only a disulfide bond between the first (Cys118) and second (Cys197) extracellular loops, which confines a helical bundle together with the N-terminus adjacent to the third extracellular loop, creating the structural organization critical for ligand binding and therefore for receptor signaling. PMID- 12081480 TI - Identification of the principal binding site for RGD-containing ligands in the alpha(V)beta(3) integrin: a photoaffinity cross-linking study. AB - By superimposing data obtained by photo-cross-linking RGD-containing ligands to the human alpha(V)beta(3) integrin onto the crystal structure of the ectopic domain of this receptor (Xiong et al. (2001) Science 294, 339-345), we have identified the binding site for the RGD triad within this integrin. We synthesized three novel analogues of the 49-amino acid disintegrin, echistatin: [Bpa(21),Leu(28)]-, [Bpa(23),Leu(28)]-, and [Bpa(28)]echistatin. Each contains a photoreactive p-benzoyl-phenylalanyl (Bpa) residue in close proximity to the RGD motif which spans positions 24-26; together, the photoreactive positions flank the RGD motif. The analogues bind with high affinity to the purified recombinant alpha(V)beta(3) integrin, but very poorly to the closely related human alpha(IIb)beta(3) platelet integrin. While echistatin analogues containing Bpa in either position 23 or 28 cross-link specifically and almost exclusively to the beta(3) subunit of alpha(V)beta(3), [Bpa(21),Leu(28)]echistatin cross-links to both the alpha(V) and the beta(3) subunits, with cross-linking to the former favored. [Bpa(23),Leu(28)]echistatin cross-links 10-30 times more effectively than the other two analogues. We identified beta(3)[109-118] as the domain that encompasses the contact site for [Bpa(28)]echistatin. This domain is included in beta(3)[99-118] (Bitan et al. (2000) Biochemistry 39, 11014-11023), a previously identified contact domain for a cyclic RGD-containing heptapeptide with a benzophenone moiety in a position that is similar to the placement of the benzophenone in [Bpa(28)]echistatin relative to the RGD triad. Recently, we identified beta(3)[209-220] as the contact site for an echistatin analogue with a photoreactive group in position 45, near the C-terminus of echistatin (Scheibler et al. (2001) Biochemistry 40, 15117-14126). Taken together, these results support the hypothesis that the very high binding affinity of echistatin to alpha(V)beta(3) results from two distinct epitopes in the ligand, a site including the RGD triad and an auxiliary epitope at the C-terminus of echistatin. Combining our results from photoaffinity cross-linking studies with data now available from the recently published crystal structure of the ectopic domain of alpha(V)beta(3), we characterize the binding site for the RGD motif in this receptor. PMID- 12081482 TI - Mutational analysis of endonuclease V from Thermotoga maritima. AB - Endonuclease V nicks damaged DNA at the second phosphodiester bond 3' to inosine, uracil, mismatched bases, or abasic (AP) sites. Alanine scanning mutagenesis was performed in nine conserved positions of Thermotoga maritima endonuclease V to identify amino acid residues involved in recognition or endonucleolytic cleavage of these diverse substrates. Alanine substitution at D43, E89, and D110 either abolishes or substantially reduces inosine cleavage activity. These three mutants gain binding affinity for binding to double-stranded or single-stranded inosine substrates in the absence of a metal ion, suggesting that these residues may be involved in coordinating catalytic metal ion(s). Y80A, H116A, and, to a lesser extent, R88A demonstrate reduced affinities for double-stranded or single stranded inosine substrates or nicked products. The lack of tight binding to a nicked inosine product accounts for the increased rate of turnover of inosine substrate since the product release is less rate-limiting. Y80A, R88A, and H116A fail to cleave AP site substrates. Their activities toward uracil substrates are in the following order: H116A > R88A > Y80A. These residues may play a role in substrate recognition. K139A maintains wild-type binding affinity for binding to double-stranded and single-stranded inosine substrate, but fails to cleave AP site and uracil substrate efficiently, suggesting that K139 may play a role in facilitating non-inosine substrate cleavage. PMID- 12081483 TI - Caught in the act: the structure of phosphorylated beta-phosphoglucomutase from Lactococcus lactis. AB - Phosphoglucomutases catalyze the interconversion of D-glucose 1-phosphate and D glucose 6-phosphate, a reaction central to energy metabolism in all cells and to the synthesis of cell wall polysaccharides in bacterial cells. Two classes of phosphoglucomutases (alpha-PGM and beta-PGM) are distinguished on the basis of their specificity for alpha- and beta-glucose-1-phosphate. beta-PGM is a member of the haloacid dehalogenase (HAD) superfamily, which includes the sarcoplasmic Ca(2+)-ATPase, phosphomannomutase, and phosphoserine phosphatase. beta-PGM is unusual among family members in that the common phosphoenzyme intermediate exists as a stable ground-state complex in this enzyme. Herein we report, for the first time, the three-dimensional structure of a beta-PGM and the first view of the true phosphoenzyme intermediate in the HAD superfamily. The crystal structure of the Mg(II) complex of phosphorylated beta-phosphoglucomutase (beta-PGM) from Lactococcus lactis has been determined to 2.3 A resolution by multiwavelength anomalous diffraction (MAD) phasing on selenomethionine, and refined to an R(cryst) = 0.24 and R(free) = 0.28. The active site of beta-PGM is located between the core and the cap domain and is freely solvent accessible. The residues within a 6 A radius of the phosphorylated Asp8 include Asp10, Thr16, Ser114, Lys145, Glu169, and Asp170. The cofactor Mg(2+) is liganded with octahedral coordination geometry by the carboxylate side chains of Asp8, Glu169, Asp170, and the backbone carbonyl oxygen of Asp10 along with one oxygen from the Asp8-phosphoryl group and one water ligand. The phosphate group of the phosphoaspartyl residue, Asp8, interacts with the side chains of Ser114 and Lys145. The absence of a base residue near the aspartyl phosphate group accounts for the persistence of the phosphorylated enzyme under physiological conditions. Substrate docking shows that glucose-6-P can bind to the active site of phosphorylated beta-PGM in such a way as to position the C(1)OH near the phosphoryl group of the phosphorylated Asp8 and the C(6) phosphoryl group near the carboxylate group of Asp10. This result suggests a novel two-base mechanism for phosphoryl group transfer in a phosphorylated sugar. PMID- 12081484 TI - S-nitrosothiols react preferentially with zinc thiolate clusters of metallothionein III through transnitrosation. AB - Metallothionein (MT) is a two-domain protein with zinc thiolate clusters that bind and release zinc depending on the redox states of the sulfur ligands. Since S-nitrosylation of cysteine is considered a prototypic cellular redox signaling mechanism, we here investigate the reactions of S-nitrosothiols with different isoforms of MT. MT-III is significantly more reactive than MT-I/II toward S nitrosothiols, whereas the reactivity of all three isoforms toward reactive oxygen species is comparable. A cellular system, in which all three MTs are similarly effective in protecting rat embryonic cortical neurons in primary culture against hydrogen peroxide but where MT-III has a much more pronounced effect of protecting against S-nitrosothiols, confirms this finding. MT-III is the only isoform with consensus acid-base sequence motifs for S-nitrosylation in both domains. Studies with synthetic and zinc-reconstituted domain peptides demonstrate that S-nitrosothiols indeed release zinc from both the alpha- and the beta-domain of MT-III. S-Nitrosylation occurs via transnitrosation, a mechanism that differs fundamentally from that of previous studies of reactions of MT with NO*. Our data demonstrate that zinc thiolate bonds are targets of S-nitrosothiol signaling and further indicate that MT-III is biologically specific in converting NO signals to zinc signals. This could bear importantly on the physiological action of MT-III, whose biological activity as a neuronal growth inhibitory factor is unique, and for brain diseases that have been related to oxidative or nitrosative stress. PMID- 12081485 TI - Proximity of the protein moiety of a GPI-anchored protein to the membrane surface: a FRET study. AB - GPI-anchored proteins are ubiquitous on the eukaryotic cell surface, where they are involved in a variety of functions ranging from adhesion to enzymatic catalysis. Indirect evidence suggests that the GPI anchor may hold the protein close to the plasma membrane; however, there is a lack of direct information on the proximity of the protein portion of GPI-anchored proteins to the bilayer surface. The present study uses fluorescence resonance energy transfer (FRET) to address this important problem. The GPI-anchored ectoenzyme placental alkaline phosphatase (PLAP) was purified from a plasma membrane extract of human placental microsomes without the use of butanol. The protein was fluorescently labeled at the N-terminus with 7-(dimethylamino)coumarin-4-acetic acid succinimidyl ester (DMACA-SE) or Oregon Green 488 succinimidyl ester (OG488-SE), and each was reconstituted by detergent dilution into defined lipid bilayer vesicles containing an increasing mole fraction of a fluorescent lipid probe. The fluorescence of the labeled PLAP donors was quenched in a concentration-dependent manner by the lipid acceptors. The energy transfer data were analyzed using an approach that describes FRET between a uniform distribution of donors and acceptors in an infinite plane. The distance of closest approach between the protein moiety of PLAP and the lipid-water interfacial region of the bilayer was estimated to be smaller than 10-14 A. This indicates that the protein portion of PLAP is located very close to the lipid bilayer, possibly resting on the surface. This contact may allow transmission of structural changes from the membrane surface to the protein, which could influence the behavior and catalytic properties of GPI-anchored proteins. PMID- 12081486 TI - Neuronal nitric oxide synthase ligand and protein vibrations at the substrate binding site. A study by FTIR. AB - Improvements in sensitivity and data processing of Fourier transform infrared (FTIR) spectroscopy enable it to be used to detect changes in protein structure at the atomic level. This paper reports a study of neuronal nitric oxide synthase (nNOS) by FTIR difference spectroscopy in the 1000-2500 cm(-1) range where vibrational bands of ligands, prosthetic groups, and protein and amino acid side chains are found. We have exploited the photolyzable CO compound of the ferrous heme of nNOS to produce light-induced CO photolysis difference spectra and to compare spectra after hydrogen/deuterium exchange. In (reduced) minus (reduced plus CO) difference spectra, negative bands at 1931 and 1907 cm(-1) are observed due to photolysis of multiple forms of ferrous heme-ligated CO, similar to those observed by resonance Raman spectroscopy [Wang et al. (1997) Biochemistry 36, 4595-4606]. Photolysis of the ferrous heme CO compound is accompanied by hitherto unreported changes in the 1000-2000 cm(-1) region that arise from changes of protein backbone, substrate, amino acid side chain, and cofactor vibrations. Preliminary assignments of vibrations are made on the basis of frequencies and the effects of hydrogen/deuterium exchange, and in the light of known atomic structures. PMID- 12081487 TI - Secondary structure and lipid interactions of the N-terminal segment of pulmonary surfactant SP-C in Langmuir films: IR reflection-absorption spectroscopy and surface pressure studies. AB - Pulmonary surfactant, a thin lipid/protein film lining mammalian lungs, functions in vivo to reduce the work of breathing and to prevent alveolar collapse. Analogues of two hydrophobic surfactant proteins, SP-B and SP-C, have been incorporated into therapeutic agents for respiratory distress syndrome, a pathological condition resulting from deficiency in surfactant. To facilitate rational design of therapeutic agents, a molecular level understanding of lipid interaction with surfactant proteins or their analogues in aqueous monolayer films is necessary. The current work uses infrared reflection-absorption spectroscopy (IRRAS) to determine peptide conformation and the effects of S palmitoylation on the lipid interactions of a synthetic 13 residue N-terminal peptide [SP-C13(palm)(2)] of SP-C, in mixtures with 1,2 dipalmitoylphosphatidylcholine (DPPC) or 1,2-dipalmitoylphosphatidylglycerol (DPPG). Two Amide I' features, at approximately 1655 and approximately 1639 cm( 1) in the peptide IRRAS spectra, are assigned to alpha-helical peptide bonds in hydrophobic and aqueous environments, respectively. In binary DPPC/SP C13(palm)(2) films, the proportion of hydrated/hydrophobic helix increases reversibly with surface pressure (pi), suggestive of the peptide being squeezed out from hydrophobic regions of the monolayer. No such effect was observed for DPPG/peptide monolayers, indicative of stronger, probably electrostatic, interactions. Depalmitoylation produced a weakened interaction with either phospholipid as deduced from IRRAS spectra and from pi-area isotherms. S Palmitoylation may modulate peptide hydration and conformation in the N-terminal region of SP-C and may thus permit the peptide to remain in the film at the high surface pressures present during lung compression. The unique capability of IRRAS to detect the surface pressure dependence of protein or peptide structure/interactions in a physiologically relevant model for surfactant is clearly demonstrated. PMID- 12081488 TI - The effects of hydrophobic mismatch between phosphatidylcholine bilayers and transmembrane alpha-helical peptides depend on the nature of interfacially exposed aromatic and charged residues. AB - In this study, we investigated the extent to which different aromatic and positively charged side chains, which often flank transmembrane segments of proteins, can influence lipid-peptide interactions. Model systems consisting of phosphatidylcholine and hydrophobic alpha-helical peptides with different flanking residues were investigated. The peptides were incorporated in relatively thick and in relatively thin lipid bilayers to create a peptide-bilayer hydrophobic mismatch, and the compensating effects on lipid structure were analyzed. When relatively long with respect to the thickness of the bilayer, the peptides that are flanked by the aromatic side chains, Trp, Tyr, and Phe, all induce a significant ordering of the lipid acyl chains, while the peptides flanked by the charged residues Lys, Arg, and His do not. However, when the peptides are relatively short with respect to the thickness of the bilayer, their effect on lipid organization does not depend primarily on their aromatic or charged character. Peptides flanked by Trp, Tyr, Lys, or (at low pH) His residues are effective in inducing mismatch-relieving cubic and inverted hexagonal phases, while analogues flanked by Phe, Arg, or (at neutral pH) His residues cannot induce an inverted hexagonal phase. The different responses to mismatch might reflect the different interfacial affinities of the residues that were investigated. PMID- 12081489 TI - Pathway of oxidative folding of alpha-lactalbumin: a model for illustrating the diversity of disulfide folding pathways. AB - The pathway of oxidative folding of alpha-lactalbumin (alpha LA) (four disulfide bonds) has been characterized by structural and kinetic analysis of the acid trapped folding intermediates. In the absence of calcium, oxidative folding of alpha LA proceeds through highly heterogeneous species of one-, two-, three-, and four-disulfide (scrambled) intermediates to reach the native structure. In the presence of calcium, the folding intermediates of alpha LA comprise two predominant isomers (alpha LA-IIA and alpha LA-IIIA) adopting exclusively native disulfide bonds, including the two disulfide bonds (Cys(61)-Cys(77) and Cys(73) Cys(91)) located within the beta-sheet calcium binding domain. alpha LA-IIA is a two-disulfide species consisting of Cys(61)-Cys(77) and Cys(73)-Cys(91) disulfide bonds. alpha LA-IIIA contains Cys(61)-Cys(77), Cys(73)-Cys(91), and Cys(28) Cys(111) disulfide bonds. The underlying mechanism of the contrasting folding pathways of calcium-bound and calcium-depleted alpha LA is congruent with the cause of diversity of disulfide folding pathways observed among many well characterized three-disulfide proteins, including bovine pancreatic trypsin inhibitor and hirudin. Our study also reveals novel aspects of the folding mechanism of alpha LA that have not been described previously. PMID- 12081490 TI - Nature of the displaceable heme-axial residue in the EcDos protein, a heme-based sensor from Escherichia coli. AB - The EcDos protein belongs to a group of heme-based sensors that detect their ligands with a heme-binding PAS domain. Among these various heme-PAS proteins, EcDos is unique in having its heme iron coordinated at both axial positions to residues of the protein. To achieve its high affinities for ligands, one of the axial heme-iron residues in EcDos must be readily displaceable. Here we present evidence from mutagenesis, ligand-binding measurements, and magnetic circular dichroism, resonance Raman, and electron paramagnetic resonance spectroscopies about the nature of the displaceable residue in the heme-PAS domain of EcDos, i.e., EcDosH. The magnetic circular dichroism spectra in the near-infrared region establish histidine-methionine coordination in met-EcDos. To determine whether in deoxy-EcDos coordination of the sixth axial position is also to methionine, methionine 95 was substituted with isoleucine. This substitution caused the ferrous heme iron to change from an exclusively hexacoordinate low-spin form (EcDosH) to an exclusively pentacoordinate high-spin form (M95I EcDosH). This was accompanied by a modest acceleration of the dissociation rates of ligands but a dramatic increase (60-1300-fold) in the association rate constants for binding of O(2), CO, and NO. As a result, the affinity for O(2) was enhanced 10-fold in M95I EcDosH, but the partition constant M = [K(d)(O(2))/K(d)(CO)] between CO and O(2) was raised to about 30 from the extraordinarily low EcDosH value of 1. Thus a major consequence of the increased O(2) affinity of this sensor was the loss of its unusually strong ligand discrimination. PMID- 12081491 TI - Activation of horse PLRP2 by bile salts does not require colipase. AB - Although structurally similar to pancreatic lipase (PL), the key enzyme of intestinal fat digestion, pancreatic lipase-related protein type 2 (PLRP2) differs from PL in certain functional properties. Notably, PLRP2 has a broader substrate specificity than PL, and unlike that of PL, its activity is not restored by colipase in the presence of bile salts. In the studies presented here, the activation mechanism of horse PLRP2 was studied through active site directed inhibition experiments, and the results demonstrate fundamental differences with that of PL. The opening of the horse PLRP2 flap occurs as soon as bile salt monomers are present, is accelerated in the presence of micelles, and does not require the presence of colipase. Moreover, in contrast to PL, horse PLRP2 is able to directly interact with a bile salt micelle to form an active binary complex, without the micelle being presented by colipase, as evidenced by molecular sieving experiments. These findings, together with the sensitivity of the horse PLRP2 flap to partial proteolysis, are indicative of a higher flexibility of the flap of horse PLRP2 relative to PL. From these results, it can be concluded that PLRP2 can adopt an active conformation in the intestine, which could be important for the further understanding of the physiological role of PLRP2. Finally, this work emphasizes the essential role of colipase in lipase catalysis at the lipid-water interface in the presence of bile. PMID- 12081492 TI - Carrier protein recognition in siderophore-producing nonribosomal peptide synthetases. AB - Nonribosomal peptide synthetases (NRPSs) use phosphopantetheine (pPant) bearing carrier proteins to chaperone activated aminoacyl and peptidyl intermediates to the various enzymes that effect peptide synthesis. Using components from siderophore NRPSs that synthesize vibriobactin, enterobactin, yersiniabactin, pyochelin, and anguibactin, we examined the nature of the interaction of such cofactor-carrier proteins with acyl-activating adenylation (A) domains. While VibE, EntE, and PchD were all able to utilize "carrier protein-free" pPant derivatives, the pattern of usage indicated diversity in the binding mechanism, and even the best substrates were down at least 3 log units relative to the native cofactor-carrier protein. When tested with four noncognate carrier proteins, EntE and VibE differed both in the range of substrate utilization efficiency and in the distribution of the efficiencies across this range. Correlating sequence alignments to kinetic efficiency allowed for the construction of eight point mutants of VibE's worst substrate, HMWP2 ArCP, to the corresponding residue in its native VibB. Mutants S49D and H66E combined to increase activity 6.2-fold and had similar enhancing effects on the downstream condensation domain VibH, indicating that the two NRPS enzymes share carrier protein recognition determinants. Similar mutations of HMWP2 ArCP toward EntB had little effect on EntE, suggesting that the position of recognition determinants varies across NRPS systems. PMID- 12081493 TI - Role of protein flexibility in the catalytic cycle of p-hydroxybenzoate hydroxylase elucidated by the Pro293Ser mutant. AB - Proline 293 of p-hydroxybenzoate hydroxylase from Pseudomonas aeruginosa is in a highly conserved region of the flavoprotein aromatic hydroxylases. It is thought to impart rigidity to the backbone, as it partially cradles the FAD in these hydroxylases. Thus, this residue has been substituted with serine by site directed mutagenesis to investigate the importance of flexibility of the peptide segment in catalysis. Differential scanning calorimetry demonstrated that the mutation has decreased the stability of the folded mutant protein compared to the wild-type PHBH. The increased flexibility in the protein backbone enhanced the accessibility of the flavin hydroperoxide intermediate to the solvent, causing an increase in the elimination of H(2)O(2) from this labile intermediate and, consequently, a decrease in the efficiency of substrate hydroxylation. Additionally, the increased accessibility of this mutant form of the enzyme makes it more susceptible than the wild-type enzyme to being trapped in the hydroxyflavin intermediate form in the presence of high levels of p hydroxybenzoate. The mutation also lowers the pK(a) of the phenolic oxygen of bound p-hydroxybenzoate, and eliminates the pH dependence of the rate constant for flavin reduction by NADPH. These experimental observations lead to a model that explains how the wild-type protein can sense the charge of the 4-substituent of the aromatic ligand and link this charge to a flavin conformational change that is required for reaction with NADPH: (i) The peptide oxygen of Pro 293 is repelled by the negative charge of the phenolic oxygen of p-hydroxybenzoate. (ii) This repulsion is transmitted through the peptide backbone, causing the movement of Asn 300. (iii) The change in the position of Asn 300 triggers the movement of the flavin from the largely buried "in" conformation to the exposed, reactive "out" conformation. PMID- 12081494 TI - Selective inhibition of the collagenolytic activity of human cathepsin K by altering its S2 subsite specificity. AB - The primary specificity of papain-like cysteine proteases (family C1, clan CA) is determined by S2-P2 interactions. Despite the high amino acid sequence identities and structural similarities between cathepsins K and L, only cathepsin K is capable of cleaving interstitial collagens in their triple helical domains. To investigate this specificity, we have engineered the S2 pocket of human cathepsin K into a cathepsin L-like subsite. Using combinatorial fluorogenic substrate libraries, the P1-P4 substrate specificity of the cathepsin K variant, Tyr67Leu/Leu205Ala, was determined and compared with those of cathepsins K and L. The introduction of the double mutation into the S2 subsite of cathepsin K rendered the unique S2 binding preference of the protease for proline and leucine residues into a cathepsin L-like preference for bulky aromatic residues. Homology modeling and docking calculations supported the experimental findings. The cathepsin L-like S2 specificity of the mutant protein and the integrity of its catalytic site were confirmed by kinetic analysis of synthetic di- and tripeptide substrates as well as pH stability and pH activity profile studies. The loss of the ability to accept proline in the S2 binding pocket by the mutant protease completely abolished the collagenolytic activity of cathepsin K whereas its overall gelatinolytic activity remained unaffected. These results indicate that Tyr67 and Leu205 play a key role in the binding of proline residues in the S2 pocket of cathepsin K and are required for its unique collagenase activity. PMID- 12081495 TI - Mechanism-based inhibitors of CD38: a mammalian cyclic ADP-ribose synthetase. AB - The soluble domain of human CD38 catalyzes the conversion of NAD(+) to cyclic ADP ribose and to ADP-ribose via a common covalent intermediate [Sauve, A. A., Deng, H. T., Angelletti, R. H., and Schramm, V. L. (2000) J. Am. Chem. Soc. 122, 7855 7859]. Here we establish that mechanism-based inhibitors can be produced by chemical stabilization of this intermediate. The compounds nicotinamide 2' deoxyriboside (1), 5-methylnicotinamide 2'-deoxyriboside (2), and pyridyl 2' deoxyriboside (3) were synthesized and evaluated as inhibitors for human CD38. The nicotinamide derivatives 1 and 2 were inhibitors of the enzyme as determined by competitive behavior in CD38-catalyzed conversion of nicotinamide guanine dinucleotide (NGD(+)) to cyclic GDP-ribose. The K(i) values for competitive inhibition were 1.2 and 4.0 microM for 1 and 2, respectively. Slow-onset characteristics of reaction progress curves indicated a second higher affinity state of these two inhibitors. Inhibitor off-rates were slow with rate constants k(off) of 1.5 x 10(-5) s(-1) for 1 and 2.5 x 10(-5) s(-1) for 2. Apparent dissociation constants K(i(total)) for 1 and 2 were calculated to be 4.5 and 12.5 nM, respectively. The similar values for k(off) are consistent with the hydrolysis of common enzymatic intermediates formed by the reaction of 1 and 2 with the enzyme. Both form covalently attached deoxyribose groups to the catalytic site nucleophile. Chemical evidence for this intermediate is the ability of nicotinamide to rescue enzyme activity after inactivation by either 1 or 2. A covalent intermediate is also indicated by the ability of CD38 to catalyze base exchange, as observed by conversion of 2 to 1 in the presence of nicotinamide. The deoxynucleosides 1 and 2 demonstrate that the chemical determinants for mechanism-based inhibition of CD38 can be satisfied by nucleosides that lack the 5'-phosphate, the adenylate group, and the 2'-hydroxyl moiety. In addition, these compounds reveal the mechanism of CD38 catalysis to proceed by the formation of a covalent intermediate during normal catalytic turnover with faster substrates. The covalent 2'-deoxynucleoside inactivators of CD38 are powerful inhibitors by acting as good substrates for formation of the covalent intermediate but are poor leaving groups from the intermediate complex because hydrolytic assistance of the 2'-hydroxyl group is lacking. The removal of the adenylate nucleophile required for the cyclization reaction provides slow hydrolysis as the only exit from the covalent complex. PMID- 12081496 TI - DNA-mediated charge transport as a probe of MutY/DNA interaction. AB - MutY is an Escherichia coli DNA repair enzyme that binds to 8-oxo-G:A and G:A mismatches and catalyzes the deglycosylation of the mismatched 2'-deoxyadenosine. We have applied DNA-mediated charge transport to probe the interaction of MutY with its DNA substrate. Oligonucleotides synthesized with a tethered rhodium intercalator and guanine doublets placed before and after the MutY binding site are used to assay for base flipping activity by MutY. On the basis of this assay, we find no evidence that MutY uses progressive base flipping as a means to find its binding site; protein binding does not perturb long-range DNA charge transport. DNA-mediated charge transport can be utilized to promote protein-DNA cross-linking from a distance. Long-range oxidation of 8-oxo-G within the MutY binding site using tethered rhodium intercalators promoted cross-linking and yielded information on MutY side chains that interact with this base. On the basis of photooxidative cross-linking of the wild type but not K142A mutant, it is evident that, within the protein complex, lysine 142 makes important contacts with 8-oxo-G. PMID- 12081497 TI - Inhibition of macrophage iNOS by selective targeting of antisense PNA. AB - Peptide nucleic acids (PNAs) are synthetic polynucleobases that bind to DNA and RNA with high affinity and specificity and with poor membrane permeability. Although PNAs have an enormous potential as antisense agents, the success of antisense PNA applications will require efficient cellular uptake. In this study, a unique antisense 14-mer anti-inducible nitric oxide synthase (iNOS) was encapsulated into erythrocytes (RBC) by hypotonic dialysis. RBC loaded with PNA (10.5 +/- 3.5 micromol/mL RBC) were targeted specifically to murine macrophages, taking advantage of an in vitro opsonization induced by ZnCl(2) and bis sulfosuccynimidil-suberate (BS(3)). This in vitro opsonization enhanced the phagocytosis of loaded RBC and the delivery of PNA into macrophages (0.72 pmol/10(6) macrophages). The efficacy of this delivery system is demonstrated by decreases in NO production and iNOS protein expression inside the macrophage. Therefore, we can suggest this novel approach for biomedical application. PMID- 12081498 TI - The stratagem utilized by the plasminogen activator from the snake Trimeresurus stejnegeri to escape serpins. AB - The plasminogen activator isolated from the venom of the snake Trimeresurus stejnegeri (TSV-PA) triggers plasmin production, along with tissue-type plasminogen activators (t-PA) and urokinase (u-PA). The half-life of TSV-PA in plasma is remarkable. We unveil in this paper two of the molecular mechanisms allowing TSV-PA to escape inhibition by plasma serpins. The first involves a phenylalanine at position 193 (chymotrypsinogen numbering system). Phe(193) distinguishes TSV-PA from nearly all trypsin-like proteinases, having glycine at this position. A mutant of TSV-PA (F193G), in which Phe(193) had been replaced by a glycine, was inactivated by plasminogen activator inhibitor 1 (PAI-1) and alpha(2)-antiplasmin 100-fold more rapidly than the wild-type enzyme. The second mechanism originates from the 37-loop of TSV-PA. Swapping the 37-loop of TSV-PA for either that of t-PA or that of u-PA also increased dramatically the rate of inactivation by PAI-1. Loop swapping and F193G mutations were additive, resulting in a rate of inactivation by PAI-1 that was 4 orders of magnitude higher than for the wild-type enzyme. The potential role of Phe(193) and of the 37-loop in the immunity of TSV-PA toward alpha(1)-antitrypsin and antithrombin is also discussed. PMID- 12081499 TI - Ca(2+) binding to both the heavy and light chains of factor VIII is required for cofactor activity. AB - Previously, we demonstrated that Ca(2+) was necessary for the generation of cofactor activity following reconstitution of factor VIII from its isolated light chain (LC) and heavy chain (HC) but that Ca(2+) did not affect HC-LC binding affinity (Wakabayashi et al. (2001) Biochemistry 40, 10293-10300). Titration of EDTA-treated factor VIII with Ca(2+) followed by factor Xa generation assay showed a two-site binding pattern, with indicated high-affinity (K(d) = 8.9 +/- 1.8 microM) and low-affinity (K(d) = 4.0 +/- 0.6 mM) sites. Analysis by equilibrium dialysis using (45)Ca and <400 microM free Ca(2+) verified a high affinity binding (K(d) = 18.9 +/- 3.7 microM). Preincubation of either HC or LC with 6 mM Ca(2+) followed by reassociation with the untreated complementary chain in the presence of 0.12 mM Ca(2+) failed to generate significant cofactor activity (<0.5 nM min(-1) (nM LC)(-1)). However, pretreatment of both HC and LC with 6 mM Ca(2+) followed by reassociation (at 0.12 mM Ca(2+)) generated high activity (7.5 +/- 0.4 nM min(-1) (nM LC)(-1)). Progress curves for activity regain following factor VIII-Ca(2+) association kinetics fitted well to a series reaction scheme rather than one of simple association (p < 0.0001), suggesting a multistep process which may include a Ca(2+)-dependent conformational change. These results suggest that factor VIII contains two Ca(2+) binding sites with different affinities and that active factor VIII can be reconstituted from HC and LC only when both chains are preactivated by Ca(2+). PMID- 12081500 TI - Cysteine biosynthetic enzymes are the pieces of a metabolic energy pump. AB - Understanding the mechanisms of free energy transfer in metabolism is fundamental to understanding how the chemical forces that sustain the molecular organization of the cell are distributed. Recent studies of molecular motors (1-3) and ATP driven proton transport (4-6) describe how chemical potential is transferred at the molecular level. These systems catalyze energy transfer through structural change and appear to be dedicated exclusively to their coupling tasks (7, 8). Here we report the discovery of a new class of energy-transfer system. It is a biosynthetic pump composed of cysteine biosynthesis enzymes, ATP sulfurylase and O-acetylserine sulfhydrylase, each with its own catalytic function and from whose interactions emerge new function: the hydrolysis of ATP. The hydrolysis is kinetically and energetically linked to the chemistry catalyzed by ATP sulfurylase, the first enzyme in the cysteine biosynthetic pathway, in such a way that each molecule of ATP hydrolyzed, each stroke of the pump, produces 1 equivalent of that enzyme's product. These findings integrate cysteine metabolism and broaden our understanding of the ways in which higher order allostery is used to effect free energy transfer. PMID- 12081501 TI - Characterization of two partially unfolded intermediates of the molecular chaperone DnaK at low pH. AB - In this study, the effect of pH on the conformation and the reactivity of the Escherichia coli Hsp70 molecular chaperone DnaK was investigated using spectroscopic and chemical assays. DnaK exhibits negligible binding of the hydrophobic dye 1-anilino-naphthalene-8-sulfonate (ANS) between pH 7 to 5.0, whereas appreciable binding occurs between pH 4.5 to 4.0. The binding of ANS to a protein is diagnostic of the presence of accessible ordered hydrophobic surfaces. Such hydrophobic surfaces are often displayed by partially folded protein intermediates such as molten globules. Nucleotide inhibits 70% of the ANS binding at pH 4.5 but none of the ANS binding at pH 4.0. Proteolysis of nucleotide-free DnaK at pH 4.5 with cathepsin D yields detectable fragments (masses > 20 kDa) of the C-terminal peptide-binding domain but none of the N-terminal ATPase domain, thus the ATPase domain is preferentially targeted for proteolysis. In contrast, proteolysis of nucleotide-free DnaK at pH 4.0 with cathepsin D cuts near the linker region, yielding both functional domains. Our interpretation of these data is that incubation of DnaK at pH 4.5 produces a partially unfolded form of the ATPase domain, in which secondary structure is mainly intact, but tertiary structure is reduced. Incubation of the protein at pH 4.0 produces an intermediate in which both functional domains have collapsed and possibly separated. Nucleotide inhibits the conformational change that occurs at pH 4.5 but not at 4.0. PMID- 12081502 TI - Kinetic characterization of the weak binding states of myosin V. AB - Myosin V is a molecular motor shown to move processively along actin filaments. We investigated the properties of the weak binding states of monomeric myosin V containing a single IQ domain (MV 1IQ) to determine if the affinities of these states are increased as compared to conventional myosin. Further, using a combination of non-hydrolyzable nucleotide analogues and mutations that block ATP hydrolysis, we sought to probe the states that are populated during ATP-induced dissociation of actomyosin. MV 1IQ binds actin with a K(d) = 4 microM in the presence of ATP gamma S at 50 mM KCl, which is 10-20-fold tighter than that of nonprocessive class II myosins. Mutations within the switch II region trapped MV 1IQ in two distinct M.ATP states with very different actin binding affinities (K(d) = 0.2 and 2 microM). Actin binding may change the conformation of the switch II region, suggesting that elements of the nucleotide binding pocket will be in a different conformation when bound to actin than is seen in any of the myosin crystal structures to date. PMID- 12081503 TI - Kinetics and pathways of charge recombination in photosystem II. AB - The mechanism of charge recombination of the S(2)Q(A)(-) state in photosystem II was investigated by modifying the free energy gap between the quinone acceptor Q(A) and the primary pheophytin acceptor Ph. This was done either by changing the midpoint potential of Ph (using mutants of the cyanobacterium Synechocystis with a modified hydrogen bond to this cofactor), or that of Q(A) (using different inhibitors of the Q(B) pocket). The results show that the recombination rate is dependent on the free energy gap between Ph and Q(A), which confirms that the indirect recombination pathway involving formation of Ph(-) has a significant contribution. In the mutant with the largest free energy gap, direct electron transfer from Q(A)(-) to P(+) predominates. The temperature dependence of the recombination rate was investigated, showing a lower activation enthalpy in this mutant compared with the WT. The data allow the determination of the rate of the direct route and of its relative weight in the various strains. The set of currently accepted values for the midpoint potentials of the Q(A)/Q(A)(-), Ph/Ph( ), and P(+)/P* couples is not consistent with the relatively rapid rate of the indirect recombination pathway found here, nor with the 3% yield of delayed fluorescence as previously estimated by de Grooth and van Gorkom (1981, Biochim. Biophys. Acta 635, 445-456). It is argued that a likely explanation is that the midpoint potentials of the two latter couples are more positive than believed due to electrostatic interactions. If such is the case, the estimation of the midpoint potential of the P(+)/P and S(2)/S(1) couples must also be revised upward, with values of 1260 and 1020 mV, respectively. PMID- 12081504 TI - Activation mechanism of CDK2: role of cyclin binding versus phosphorylation. AB - Activation of the cyclin-dependent kinases is a two-step process involving cyclin binding followed by phosphorylation at a conserved threonine residue within the kinase activation loop. In this study, we describe the separate roles of cyclin A binding versus phosphorylation in the overall activation mechanism of CDK2. Interaction of CDK2 with cyclin A results in a partially active complex that is moderately defective in the binding of the protein substrate, but not ATP, and severely defective in both phosphoryl group transfer and turnover. Alternatively, phosphorylation of the CDK2 monomer also results in a partially activated species, but one that is severely (> or = 480-fold) defective in substrate binding exclusively. Catalytic turnover in the phosphorylated CDK2 monomer is largely unimpaired (approximately 8-fold lower). Our data support a model for the activation of CDK2 in vivo, in which interaction of unphosphorylated CDK2 with cyclin A serves to configure the active site for ground-state binding of both ATP and the protein substrate, and further aligns ATP in the transition state for phosphoryl transfer. Optimizing the alignment of protein substrates in the phosphoryl transfer reaction is the principal role of phosphorylation at Thr(160). PMID- 12081506 TI - Patterns of computer usage among medical practitioners in rural and remote Queensland. AB - As part of a more detailed needs analysis, patterns of computer usage among medical practitioners in rural and remote Queensland were investigated. Utilising a questionnaire approach, a response rate of 23.82% (n = 131) was obtained. Results suggest that medical practitioners in rural and remote Queensland are relatively sophisticated in their use of computer and information technologies and have embraced computerisation to a substantially higher extent compared with their urban counterparts and previously published estimates. Findings also indicate that a substantial number of rural and remote practitioners are utilising computer and information technologies for clinical purposes such as pathology, patient information sheets, prescribing, education, patient records and patient recalls. Despite barriers such as bandwidth limitations, cost and the sometimes unreliable quality of Internet service providers, a majority of rural and remote respondents rated an Internet site with continuing medical education information and services as being important or very important. Suggestions that "rural doctors are slow to adapt to new technologies" are questioned, with findings indicating that rural and remote medical practitioners in Queensland have adapted to, and utilise, information technology to a far higher extent than has been previously documented. PMID- 12081507 TI - Developing strategies to gather information about the maternity experiences of indigenous women in an acute care setting. AB - This project emerged from the concerns of health care professionals at a large acute care hospital in the Northern Territory of Australia. The aim of the project was to develop educational resources through information and experiences provided by Aboriginal and Torres Strait Islander women and health care professionals. The objectives are to optimise collaboration and participation by Indigenous and Torres Strait Islander women in sharing their maternity experiences about the birthing of their baby either in hospital, or out of hospital before arrival. Stage one of a three stage project is reported here. In stage one a modified Delphi method was used to interview ten purposefully chosen professional Indigenous women with insight into the research process and changing social arrangements. Stage two involved the development of an interview schedule and face-to-face interviews. Stage three includes the development of a questionnaire to be answered by health care professionals. Discussion of the background to the study, reviewed literature and issues identified from interviews with the professional Indigenous women regarding best methods and appropriate data collection processes is presented. Research described here explores issues of concern for Indigenous women from the Northern Territory around their child birthing experiences in an acute care setting. PMID- 12081505 TI - Characterization of Ca2+/calmodulin-dependent protein kinase I as a myosin II regulatory light chain kinase in vitro and in vivo. AB - Ca(2+)/calmodulin (CaM)-dependent protein kinase I (CaM-KI), which is a member of the multifunctional CaM-K family, is thought to be involved in various Ca(2+) signalling pathways. In this report, we demonstrate that CaM-KI activated by an upstream kinase (CaM-K kinase), but not unactivated CaM-KI, phosphorylates myosin II regulatory light chain (MRLC) efficiently ( K (cat), 1.7 s(-1)) and stoichiometrically (approximately 0.8 mol of phosphate/mol) in a Ca(2+)/CaM dependent manner in vitro. One-dimensional phosphopeptide mapping and mutational analysis of MRLC revealed that the activated CaM-KI monophosphorylates only Ser 19 in MRLC. Transient expression of the Ca(2+)/CaM-independent form of CaM-KI (CaM-KI(1-293)) in HeLa cells induced Ser-19 phosphorylation of myosin, II accompanied by reorganization of actin filaments in the peripheral region of the cells. CaM-KI-induced reorganization of actin filaments was suppressed by co expression of non-phosphorylatable MRLC mutants (S19A and T18AS19A). Furthermore, a kinase-negative form of CaM-KI (CaM-KI(1-293,K49E)) significantly reduced reorganization of actin filaments, indicating a dominant negative effect. This is the first demonstration that the activation of the CaM-KI cascade induces myosin II phosphorylation, resulting in regulation of actin filament organization in mammalian cells. PMID- 12081508 TI - The maternity experiences of indigenous women admitted to an acute care setting. AB - This is the report of stage two of a three-stage project. The aim of the project is to develop educational resources through information and experiences provided by Aboriginal and Torres Strait Islander women and health care professionals. The objectives were to optimise collaboration and participation by Indigenous and Torres Strait Islander women in sharing their maternity experiences about the birthing of their babies either in hospital, or out of hospital before arrival. The descriptive research of stage two explores issues that existed for twelve Indigenous women during and after their birthing experiences in an acute care setting. The results will inform patient and professional educational programs and policy in order to enhance the experiences for Indigenous women admitted to the acute care setting before or following birth. The results show broad variation in responses related to antenatal care, preparation for hospital and for birth, coming into town, accommodation, family support, experiences while in hospital, relationship with hospital staff and being away from home. Miscommunication and lack of cultural and spiritual understanding by health care professionals generally is a constant theme. The need for preparation of Indigenous women generally, prior to admission to an acute care setting, is confirmed. PMID- 12081510 TI - Challenges to rural medical education: a student perspective. AB - The key feature of the Greater Murray Clinical School model is the attachment of students to patients. Students follow their patients through the health care system, in contrast to the standard approach where students are attached to doctors or specialty based clinical units. The patient/student coupling occurs at the primary care level, which mostly but not exclusively will occur in the GP's office. Students anchor their knowledge by seeing the natural progression of common illnesses, the impact of behavioural aspects on health and disease, and by experiencing continuity of care. Along their path they develop good problem solving skills and learn to understand the health care system they will become part of. The main obstacle in teaching a medical undergraduate curriculum in a rural setting is that large geographical distances separate students, teachers and resources. Consequently, information technology will play an important role in terms of delivery of the GMCS curriculum. Moreover, there is potential for flow-on benefits to the community following integration of new information technology into the local health infrastructure. PMID- 12081509 TI - Opinions of healthcare professionals regarding the maternity experiences of indigenous women in an acute care setting. AB - This is the report of stage three of a three stage project. The aims of the overall project were to seek advice from expert Indigenous women on how to gather information from birthing Indigenous women while in hospital. Information was gathered from Indigenous women who gave birth in hospital or were admitted after the birth of their baby. The opinions of healthcare professionals on the care of these women were investigated. This paper describes the opinions and concerns of healthcare professionals regarding identified cultural, social and economical issues for Indigenous women at a large acute care hospital in the Northern Territory, Australia. The opinions of healthcare professionals are vital if the needs and concerns of Indigenous women are to be considered within the hospital setting. Stages one and two of this project have been presented previously. Stage three, reported here, describes a survey of 18 non-Indigenous and one Indigenous healthcare professionals regarding their opinions of the maternity experiences of Indigenous women admitted to an acute care setting. The consultative process for designing the questionnaire and reflections on issues raised by Indigenous women during their interviews in the previous two stages is presented. The methodology of the survey, demographics and perceptions of healthcare professionals who participated in the project are examined and the similarities and differences between the issues raised by the healthcare professionals and the Indigenous women in stage two of the project are discussed. Suggestions made by the healthcare professionals regarding improving quality of care for Indigenous women when admitted to an acute healthcare setting are presented. PMID- 12081511 TI - Strengthening primary health care: building the capacity of rural communities to access health funding. AB - Present health funding models can place onerous pressures on rural health services. Staff may lack the time, resources, access to data, and the expertise needed to complete complex and lengthy funding submissions. This present study describes an innovative capacity-building approach to working with Victorian rural communities seeking to access health care funding through the Regional Health Services Program. This approach used several strategies: engaging stakeholders in targeted rural communities, developing an information kit and running a workshop on preparing submissions to the Regional Health Services Program, facilitating community consultations, and providing ongoing support with submissions. Six rural communities were supported in this way. Four have been funded to date, with a combined annual recurrent budget for new primary health care services of over $2.5 million. Each community has developed a service delivery model that meets the particular needs of their local area. This capacity building approach is both effective and replicable to other health funding opportunities. PMID- 12081512 TI - Why nurses are attracted to rural and remote practice. AB - This is the second paper of a larger study that examined the factors influencing the decisions of rural and remote area nurses, formerly employed by Queensland Health, to leave or to remain in this area of nursing. The study was a cross sectional survey that gathered data from nurses who had resigned from permanent positions in Queensland Health during the period February 1999 to May 2000. This paper reports only those factors that influenced their decision to remain in rural and remote area practice, which can be categorised into personal, professional and rural influences. The results of this study are congruent with previous national and international research findings into these issues for nurses in rural and remote areas. The paper makes recommendations to attract nurses to the rural and remote work force. PMID- 12081513 TI - The role of computational fluid dynamics for artificial organ design. PMID- 12081514 TI - Computational fluid dynamics and vascular access. AB - Anastomotic intimal hyperplasia caused by unphysiological hemodynamics is generally accepted as a reason for dialysis access graft occlusion. Optimizing the venous anastomosis can improve the patency rate of arteriovenous grafts. The purpose of this study was to examine, evaluate, and characterize the local hemodynamics and, in particular, the wall shear stresses in conventional venous end-to-side anastomosis and in patch form anastomosis (Venaflo) by three dimensional computational fluid dynamics (CFD). We investigated the conventional form of end-to-side anastomosis and a new patch form by numerical simulation of blood flow. The numerical simulation was done with a finite volume-based algorithm. The anastomotic forms were constructed with usual size and fixed walls. Subdividing the flow domain into multiple control volumes solved the fundamental equations. The boundary conditions were identical for both forms. The velocity profile of the patch form is better than that for the conventional form. The region of high static pressure caused by flow stagnation is reduced on the vein floor. The anastomotic wall shear stress is decreased. The results of this study strongly support patch form use to reduce the incidence of intimal hyperplasia and venous anastomotic stenoses. PMID- 12081515 TI - Numerical calculation of hemolysis levels in peripheral hemodialysis cannulas. AB - Hemolysis in extracorporeal life support systems presents an underestimated problem. In this article, we investigate the hemolytic potential of peripheral hemodialysis cannulas numerically. An axisymmetrical finite element model of 3 cannula sizes was built (13G, 14G, and 16G) that was refined sufficiently in the vicinity of the cannula tip to compute accurately scalar shear stresses. Scalar shear stresses were utilized in Giersiepen's equation to calculate the red blood cell damage (RBD) along streamlines. The streamlines were chosen such that they bound a percentage of the blood flow through the cannula. By integration of the RBD results, the total damage of the cannula was determined and expressed in modified index of hemolysis for comparison with published results. Calculated RBD was overestimated by Giersiepen's equation. The ranking of the cannulas according to their hemolytic potential was preserved. This indicates that power-law equations may be suited for hemolysis prediction of laminar flow devices. PMID- 12081516 TI - Contributions of a finite element model for the geometric optimization of an implantable bioartificial pancreas. AB - The extravascular implantation of islets of Langerhans immunoprotected within a permselective membrane is a promising method to treat diabetes mellitus. However, oxygen limitation due to purely diffusive solute transport was considered to provoke tissue necrosis and graft failure. We built a solute transport model based on a finite element method aiming at optimizing the hollow fiber geometry. With a low islet density, the influence of oxygen axial flux inside the fiber was underlined and a characteristic length for oxygen supply was introduced. This study allowed the conclusion that islet density must be adapted to the fiber diameter chosen for implantation. PMID- 12081517 TI - Computational flow modeling in hollow-fiber dialyzers. AB - A three-dimensional finite volume model of the blood-dialysate interface over the complete length of the dialyzer was developed. Different equations govern dialyzer flow and pressure distribution (Navier-Stokes) and radial transport (Darcy). Blood was modeled as a non-Newtonian fluid with a viscosity varying in radial and axial direction determined by the local hematocrit, the diameter of the capillaries, and the local shear rate. The dialysate flow was assumed to be an incompressible, isothermal laminar Newtonian flow with a constant viscosity. The permeability characteristics of the membrane were calculated from laboratory tests for forward and backfiltration. The oncotic pressure induced by the plasma proteins was implemented as well as the reduction of the overall permeability caused by the adhesion of proteins to the membrane. From the calculated pressure distribution, the impact of flow, hematocrit, and capillary dimensions on the presence and localization of backfiltration can be investigated. PMID- 12081518 TI - Predicting membrane oxygenator pressure drop using computational fluid dynamics. AB - Three-dimensional computational fluid dynamic (CFD) simulations of membrane oxygenators should allow prediction of spatially dependent variables and subsequent shape optimization. Fiber bed complexity and current computational limitations require the use of approximate models to predict fiber drag effects in complete device simulations. A membrane oxygenator was modified to allow pressure measurement along the fiber bundle in all cardinal axes. Experimental pressure drop information with water perfusion was used to calculate the permeability of the fiber bundle. A three-dimensional CFD model of a commercial membrane oxygenator was developed to predict pressure drops throughout the device. Darcy's Law was used to account for the viscous drag of the fibers and was incorporated as a momentum loss term in the conservation equations. Close agreement was shown between experimental and simulated pressure drops at lower flow rates, but the simulated pressure drops were lower than experimental results at higher flows. Alternate models of fiber drag effects and flow field visualization are suggested as means to potentially improve the accuracy of the flow simulation. Computational techniques coupled with experimental verification offer insight into model validity and show promise for the development of accurate three-dimensional simulations of membrane oxygenators. PMID- 12081519 TI - Computational fluid dynamics insights in the design of mechanical heart valves. AB - Computational fluid dynamics (CFD) analysis can provide detailed, three dimensional predictions of blood flow through mechanical heart valves, which can help to optimize valve hemodynamics and reduce the potential for blood clotting. A number of CFD studies, considering both forward and retrograde flow through valves, have been published. In this paper, a geometrically accurate CFD model capable of predicting the three-dimensional, time-dependent flow through an open ATS bileaflet valve is presented. A detailed picture of the blood flow is obtained, including small-scale flow features in the pivot regions. Results from the model can also be used to investigate the opening position of the ATS valve leaflets. Future work will be aimed toward improved models that provide valuable design information while minimizing the development time and computational resources required. Such practical CFD models clearly have the potential to reduce the costs, time scales, and risks associated with development of new heart valve designs. PMID- 12081520 TI - Computational fluid dynamics and stent design. AB - Stents are small, usually metallic tubes that are intended to prop open arteries blocked with atherosclerotic plaques. While stents have been used successfully in recent years, they still suffer from failure due to development of new tissue in stented segment (restenosis). Variations in the failure rates associated with different stent designs have led researchers to investigate the role of near-wall flow patterns. While there is no direct evidence yet, the patterns of flow stagnation as the blood flows past the stent struts may affect the restenosis process. Computational fluid dynamics (CFD) approaches are well suited for obtaining detailed information on stent flow patterns. Many CFD simulations make use of a two-dimensional model. The strong dependence of flow stagnation on stent strut spacing has been clearly demonstrated. These results have been employed to interpret the results of in vitro experiments designed to elucidate the mechanisms of restenosis. PMID- 12081521 TI - Validation of the coupling of magnetic resonance imaging velocity measurements with computational fluid dynamics in a U bend. AB - Magnetic resonance imaging (MRI) can be used in vivo in combination with computational fluid dynamics (CFD) to derive velocity profiles in space and time and accordingly, pressure drop and wall shear stress distribution in natural or artificial vessel segments. These hemodynamic data are difficult or impossible to acquire directly in vivo. Therefore, research has been performed combining MRI and CFD for flow simulations in flow phantoms, such as bends or anastomoses, and even in human vessels such as the aorta, the carotid, and the abdominal bifurcation. There is, however, no unanimity concerning the use of MRI velocity measurements as input for the inflow boundary condition of a CFD simulation. In this study, different input possibilities for the inflow boundary conditions are compared. MRI measurements of steady and pulsatile flow were performed on a U bend phantom, representing the aorta geometry. PAMFLOW (ESI Software, Krimpen aan den Ussel, The Netherlands), an industrial CFD software package, was used to solve the Navier-Stokes equations for incompressible flow. Three main parameters were found to influence the choice of an inflow boundary condition type. First, the flow rate through a vessel should be exact, since it proves to be a determining factor for the accuracy of the velocity profile. The other decisive parameters are the physiology of the flow profile and the required computer processing unit time. Our comparative study indicates that the best way to handle an inflow boundary condition is to use the velocities measured by MRI at the inflow plane as being fixed velocities. However, before using these MRI velocity data, they first should be corrected for the partial volume effect by filtering and second scaled in order to obtain the correct flow rate. This implies that a reliable flow rate measurement absolutely is needed for CFD calculations based on MRI velocity measurements. PMID- 12081522 TI - An infection-preventing bilayered collagen membrane containing antibiotic-loaded hyaluronan microparticles: physical and biological properties. AB - An infection-preventing bilayered membrane consisting of a dense and porous collagen membrane has been developed. The membrane was fabricated using a combined freeze-drying/air-drying method. Hyaluronan (HA) microparticles containing silver sulfadiazine (AgSD) were fabricated by gelling an HA solution with calcium chloride and were incorporated into collagen layers to allow the sustained release of AgSD. In vitro biodegradability of the membrane and the release of AgSD from the membrane could be controlled by cross-linking the membrane with ultraviolet (UV) irradiation. In a cytotoxicity test, cellular damage was minimized by the sustained release of AgSD from dressings. The antibacterial capacity of the material against Pseudomonas aeruginosa was investigated using the Bauer-Kirby disk diffusion test, and bacterial growth was found to be inhibited for 4 days. In vivo tests showed that the bilayered membrane was associated with greater tissue regeneration than a polymeric membrane and with no infection-related biological signs. PMID- 12081523 TI - Evaluation of cellular affinity and compatibility to biodegradable polyesters and Type-II collagen-modified scaffolds using immortalized rat chondrocytes. AB - Immortalized rat chondrocytes (IRCs) were employed to evaluate the cytocompatibility of different biodegradable polyester scaffolds for chondrocyte seeding and cartilage tissue engineering in vitro due to the limitation of using freshly harvested chondroctyes. Cells were seeded onto the films and the porous substrates as well as into the three-dimensional scaffolds made of the biodegradable polyesters including poly(l-lactide) (PLLA) and two poly(lactide-co glycolide)s (PLGAs). The materials were characterized by water contact angle, electron spectroscopy for chemical analysis (ESCA), and microscopy. PLGA50/50, one of the PLGAs, had the largest cell numbers at 24 h and 96 h (close to the tissue culture polystyrene control), possibly due to its lower contact angle, higher oxygen/carbon (O/C) atomic ratio, and larger degradation rate. When the surface was further modified by cross-linked Type-II collagen, cell population was significantly enhanced (two- to fourfold). The adhesion and proliferation behavior of IRCs on different materials was parallel to that of rabbit chondrocytes, but was more reproducible in general. IRCs are thus suitable for evaluation of different polymer scaffolds. Despite the favorable cytocompatibility of PLGA50/50, blending with a small portion of PLLA is required for easy fabrication and collagen modification. Scaffolds made of blended materials by freeze-drying procedure with the surface modified by cross-linked Type-II collagen were demonstrated as the ideal templates for chondrocyte seeding in our study. PMID- 12081525 TI - The tomato ethylene receptor gene family: Form and function. AB - Phytohormones are essential for integrating many aspects of plant development and responses to the environment. Regulation of hormonally controlled events occurs at multiple levels: synthesis, catabolism and perception (Trewavas 1983, Bradford and Trewavas 1994). At the level of perception, sensitivity to hormones can be regulated both spatially and temporally during the life cycle. An example of spatial regulation is the differential response to a hormone that occurs during organ abscission. Temporally, sensitivity of an organ to a hormone may change during maturation, as occurs during fruit ripening. In this review, we will focus on the initial event in recognition of one hormone, ethylene. The ethylene receptor was the first plant hormone receptor to be unambiguously identified. Over the last few years, great progress has been made in elucidating the genes involved in ethylene action. Nonetheless, the mechanisms of signal transduction remain to be established. Here, we will address the status of the tomato receptor gene family and the evidence that regulation of receptor gene expression can influence the response of the plant to the hormone. PMID- 12081524 TI - Networking of phospholipases in plant signal transduction. AB - Phospholipases are activated in response to various cellular and environmental cues. Their activation can affect many cellular processes through their roles in signal transduction. Recent advances in the biochemical and molecular understanding of phospholipase D (PLD) have provided insights into potential networks of PLDs and other phospholipases in plants. PLDs are a family of heterogeneous enzymes, and the activities of the multiple types of PLDs are regulated in distinctly different manners. Phosphoinositides, free fatty acids, lysophospholipids, and calcium are differential modulators of PLDs. Since these modulators are substrates, products, or downstream targets of phospholipase As and phospholipase Cs, there are many potential regulatory and metabolic interrelationships among the various PLDs and other phospholipases. PMID- 12081526 TI - From common signalling components to cell specific responses: insights from the cereal aleurone. AB - Studies into the molecules underlying plant signal transduction events continue to reveal the involvement of highly conserved factors such as Ca2+, calmodulin, cyclic GMP and phospholipases in a remarkably diverse array of physiological processes. The hormonal response systems in the aleurone cells of the cereal grain and in the stomatal guard cell are beginning to reveal how diversity of response can be hard wired into these cells despite the use of these common signalling intermediates. In both the aleurone and the guard cell ABA signalling operates through the action of phospholipase D and alterations in a Ca2+ dependent signalling system. The role of phospholipase D is highly analogous in these two divergent cell types, perhaps reflecting the closeness of this enzyme to a conserved ABA receptor. However, specificity in response becomes evident in elements downstream from PLD, such as in the Ca2+ signalling system. For example, ABA has opposite effects on cytoplasmic Ca2+ in the aleurone and guard cell. Combining the Ca2+-dependent signalling activities in networks with parallel regulatory activities such as cyclic GMP appears to underlie the flexible regulatory systems that are the hallmark of plant cell function. PMID- 12081527 TI - Isolation of photorespiratory mutants from Lotus japonicus deficient in glutamine synthetase. AB - A mutagenesis programme using ethyl methanesulphonate (EMS) was carried out on Lotus japonicus (Regel) Larsen cv. Gifu in order to isolate photorespiratory mutants in this model legume. These mutants were able to grow in a CO2-enriched atmosphere [0.7% (v/v) CO2] but showed stress symptoms when transferred to air. Among them, three mutants displayed low levels of glutamine synthetase (GS; EC 6.3.1.2) activity in leaves. The mutants accumulated ammonium in leaves upon transfer from 0.7% (v/v) CO2 to air. F1 plants of back crosses to wild type were viable in air and F2 populations segregated 3 : 1 (viable in air : air-sensitive) indicative of a single Mendelian recessive trait. Complementation tests showed that the three mutants obtained were allelic. Chromatography on DEAE-Sephacel used to separate the cytosolic and plastidic GS isoenzymes together with immunological data showed that: (1) mutants were specifically affected in the plastidic GS isoform, and (2) in L. japonicus the plastidic GS isoform eluted at lower ionic strength than the cytosolic isoform, contrary to what happens in most plants. The plastidic GS isoform present in roots of wild type L. japonicus was also absent in roots of the mutants, indicating that this plastidic isoform from roots was encoded by the same gene than the GS isoform expressed in leaf tissue. Viability of mutant plants in high-CO2 conditions indicates that plastidic GS is not essentially required for primary ammonium assimilation. Nevertheless, mutant plants did not grow as well as wild type plants in high-CO2 conditions. PMID- 12081528 TI - Carbon assimilation, partitioning and export in mature cladophylls of two asparagus (Asparagus officinalis) cultivars with contrasting yield. AB - To assess the physiological aspects underpinning cultivar difference in asparagus (Asparagus officinalis L.) yield, diel carbon exchange, carbon partitioning and export and sucrose phosphate synthase (SPS) activity were examined in mature cladophyll tissue of two asparagus cultivars (ASP-69 and ASP-03) under field conditions. Both cultivars exhibited similar diel patterns in carbon exchange rate (CER) and carbohydrate partitioning. Rates of carbon export estimated from CER and dry mass changes were the highest at midday and coincided with maximum assimilation rate. Carbon export accounted for about 74% of carbon fixation during the photoperiod. No diel fluctuations were observed in SPS activity in either cultivar. A positive correlation between day CER and carbon export rate (r2 = 0.87) was found and this relationship did not differ between the two cultivars studied. The greater carbon export rate measured in the high-yielding cultivar (ASP-69) was associated with significantly higher CER in comparison to the low-yielding cultivar (ASP-03). However, a correlation between sucrose concentration and carbon export rate did not exist. Biochemical evidence indicated that the greater CER in ASP-69 was associated with a significantly greater SPS activity (P 0.05). Phloem 14C exudate analysis revealed that 14C flux out of cladophyll tissue in ASP-69 was significantly greater than in ASP-03. These results indicate a feed-forward effect of rate of photosynthesis on assimilate export in the two cultivars studied. PMID- 12081529 TI - ABA and polyamines act independently in primary leaves of cold-stressed tomato (Lycopersicon esculentum). AB - The effects of ABA and putrescine, a polyamine, on cold-induced membrane leakage were investigated using primary leaves of wild-type and an ABA-deficient mutant, flacca, of tomato (Lycopersicon esculentum Mill.). The amount of chilling-induced electrolyte leakage from flacca leaves was much higher than that from the wild type leaves. When applied exogenously ABA reduced cold-induced electrolyte leakage from leaves of both wild-type and the flacca mutant. However, the cold induced electrolyte leakage from flacca leaves was not as pronounced as in the wild-type indicating that ABA is an important mediator in response to cold stress in the leaves. Putrescine reduced cold-induced electrolyte leakage from both wild type and flacca leaves. Synthesis of putrescine in the leaves was increased by cold treatment. DFMO, a biosynthetic inhibitor of the polyamine, increased electrolyte leakage from cold-treated leaves, and exogenously applied putrescine decreased the enhanced leakage to the control level. Therefore, this polyamine is thought also to be involved in the response to cold stress of tomato leaves. Both ABA and putrescine were protective against cold stress, but exogenously applied ABA decreased the endogenous level of putrescine in the leaves. Furthermore, the DMFO-increased electrolyte leakage in cold-stressed leaves was completely abolished by the application of ABA. These results suggest that ABA is a major regulator in the response to cold stress in tomato leaves and that it does not exert its role via putrescine in the response to cold stress. PMID- 12081530 TI - Effects of Pb-EDTA and EDTA on oxidative stress reactions and mineral uptake in Phaseolus vulgaris. AB - Sequestration of Pb by synthetic chelates has been reported to increase bioavailability, uptake, and translocation of this metal in plants. In this work the potential phytotoxic effects of Pb-EDTA were investigated in Phaseolus vulgaris L. cv. Limburgse vroege plants grown on hydroponics. Addition of 50 microM Pb-EDTA to the nutrient solution caused a significant induction of syringaldazine peroxidase (SPOD; EC 1.11.1.7) in roots and primary leaves and guaiacol peroxidase (GPOD; EC 1.11.1.7) in leaves. Addition of 100 microM Pb-EDTA further exacerbated ascorbate peroxidase (APOD; EC 1.11.1.11), GPOD, dehydroascorbate reductase (DHAR; EC 1.8.5.1), glutathione reductase (GR; EC 1.6.4.2) and malic enzyme (ME; EC 1.1.1.40) in roots and APOD and ME in primary leaves. Addition of 200 microM Pb-EDTA also induced DHAR in leaves. This induction of peroxidases (SPOD, GPOD, APOD), enzymes of the ascorbate-glutathione cycle (DHAR, GR in roots) and of an NADP+ reducing enzyme in roots and primary leaves indicates that oxidative stress has been initiated. At 200 microM Pb-EDTA, chlorophyll a and b content in leaves was significantly reduced while visible effects on root morphology and shoot length were observed, while no significant morphological effects were found in the leaves, confirming the sensitive character of the measured enzymes as plant stress indicators. Elevation of the Pb EDTA concentration in the growth medium significantly reduced the content of Ca, Fe, Mn and Zn taken up by plants, probably due to ion leakage as a result of observed toxicity. Addition of up to 200 microM EDTA increased chelation of divalent cations in nutrient solution resulting in reduced plant uptake of Zn, Cu, Fe and Mn. This did not result in phytotoxicity. PMID- 12081531 TI - The electrical impedance spectroscopy of Scots pine needles during cold acclimation. AB - The electrical impedance spectroscopy (EIS) was applied to current-year needles of Scots pine (Pinus sylvestris L.) in an 8-year provenance field trial in central Finland during frost hardening. The EIS analysis of the needles using a Model-A equivalent circuit indicated a sequence of events in the needles during their cold acclimation. Some of the EIS-parameters referred to maturation phenomena occurring during the pre-hardening phase at the end of the growing season, and some parameters displayed a clear coincidence with the frost hardening itself. Significant differences between provenances were found in several of the Model-A parameters. Extracellular resistance (re) and beta coefficient decreased in all provenances in the pre-hardening phase in August and until mid-September. In the same phase, both the intracellular resistance (ri) and the cell membrane time constant (taum) first increased and then decreased. According to taum, re and beta there was a clear gradation between provenances in the pre-hardening phase. From the end of September significant differences were found in the intracellular resistance between provenances, corresponding with the differences in their hardening pattern. The dry weight (DW) content of needles increased during the study period but no clear differences were found between the provenances. PMID- 12081532 TI - Salt stress induces up-regulation of an efficient chloroplast antioxidant system in the salt-tolerant wild tomato species Lycopersicon pennellii but not in the cultivated species. AB - The response of the chloroplastic antioxidant system of the cultivated tomato Lycopersicon esculentum (Lem) and its wild salt-tolerant related species L. pennellii (Lpa) to NaCl stress was studied. An increase in H2O2 level and membrane lipid peroxidation was observed in chloroplasts of salt-stressed Lem. In contrast, a decrease in these indicators of oxidative stress characterized chloroplasts of salt-stressed Lpa plants. This differential response of Lem and Lpa to salinity, correlates with the activities of the antioxidative enzymes in their chloroplasts. Increased activities of total superoxide dismutase (SOD), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), glutathione-S transferase (GST), phospholipid hydroperoxide glutathione peroxidase (PHGPX) and several isoforms of non-specific peroxidases (POD) were found in chloroplasts of salt-treated Lpa plants. In these chloroplasts, in contrast, activity of lipoxygenase (LOX) decreased while in those of salt-stressed Lem it increased. Although total SOD activity slightly increased in chloroplasts of salt-treated Lem plants, differentiation between SOD types revealed that only stromal Cu/ZnSOD activity increased. In contrast, in chloroplasts of salt-treated Lpa plants FeSOD activity increased while Cu/ZnSOD activity remained unchanged. These data indicate that salt-dependent oxidative stress and damage, suffered by Lem chloroplasts, was effectively alleviated in Lpa chloroplasts by the selective up regulation of a set of antioxidative enzymes. Further support for the above idea was supplied by leaf discs experiments in which pre-exposure of Lpa plants to salt-treatment conferred cross-tolerance to paraquat-induced oxidative stress while increased oxidative damage by paraquat-treatment was found in salt-stressed Lem plants. PMID- 12081533 TI - Isolation and identification of a potent allelopathic substance in rice root exudates. AB - A search for growth inhibitors in rice root exudates was undertaken in order to clarify the allelopathic system in rice (Oryza sativa L.). Rice seedlings inhibited the growth of cress (Lepidium sativum L.) and lettuce (Lactuca sativa L.) seedlings when the cress and lettuce were grown with rice seedlings. The putative compound causing the inhibitory effect of rice seedlings was isolated from their culture solution, and the chemical structure of the inhibitor was determined by spectral data as momilactone B. Momilactone B inhibited the growth of cress and lettuce seedlings at concentrations greater than 3 and 30 microM, respectively. The concentration of momilactone B was 3.4 and 1.1 nmol per seedling in the culture solutions of husked and non-husked rice seedlings, respectively. These results suggest that rice seedlings may release momilactone B into the environment and the stress caused by the husk-treatment may increase the amount of momilactone B released. Thus, momilactone B may play an important role in rice allelopathy. PMID- 12081534 TI - Water entry into detached root systems saturates with increasing externally applied pressure; a result inconsistent with models of simple passive diffusion. AB - The most widely accepted model of radial water entry from the soil into the xylem of roots is based on principles of ordinary passive diffusion. However, long standing problems with this model remain unresolved, which concern variable intrinsic properties of conductivity, Lp, changing reflection coefficients, sigma, and inaccurate resolution of osmotic differentials between the soil and xylem. Our study re-examined pressure flow relationships in isolated roots of tomato (Lycopersicon esculentum Mill. cv. Montfavet), pea (Pisum sativum cv. Baccara) and soybean (Glycine max L. Merryl cv. Essor) manipulated in a pressure chamber. In addition to problems previously recognized with the simple passive diffusion model, a new conflict, flow saturation, was observed at high pressures. Experiments revealed that the plateau in flow, Jmax seen at high pressures followed natural rhythms diurnally and developmentally, and was not due to root damage or unnatural flow restriction. Near the end of the photoperiod, Jmax closely correlated with root dry mass. The above inconsistencies between observations in pressure-flow kinetics and ordinary passive diffusion indicate that either the current model should be adjusted or a new model should be proposed. PMID- 12081535 TI - Jasmonic acid affects plant morphology and calcium-dependent protein kinase expression and activity in Solanum tuberosum. AB - The effect of jasmonic acid (JA) on plant growth and on calcium-dependent protein kinase (CDPK) activity and expression was studied in non-photoperiodic potato plants, Solanum tuberosum L. var. Spunta, grown in vitro. Stem cuttings were grown for 45 days (long treatment, LT) in MS medium with increasing concentrations of JA. For short treatments (ST) adult plants grown in MS were transferred for 1, 4 and 20 h to JA containing media. During the LT, low concentrations of JA promoted cell expansion and shoot elongation while higher concentrations caused growth inhibition. Under these conditions, treated plants showed root shortening and tuber formation was not induced. Morphological and histochemical studies using light microscopy and TEM analysis of leaves from treated plants revealed that JA also affected subcellular organelles of mesophyll cells. Peroxisomes increased in size and number, and an autophagic process was triggered in response to high concentrations of the hormone. CDPK activity, determined in crude extracts of treated plants (LT), was inhibited (up to 80%). Plant growth and CDPK inhibition were reverted upon transfer of the plants to hormone-free medium. Soluble CDPK activity decreased in response to JA short treatment. Concomitantly, a decline in the steady state levels of StCDPK2 mRNA, a potato CDPK isoform that is expressed in leaves, was observed. These data suggest that the phytohormone down-regulated the expression and activity of the kinase. PMID- 12081536 TI - Abscisic acid, phaseic acid and gibberellin contents associated with dormancy and germination in barley. AB - Analyses of abscisic acid (ABA), ent-kaurenoids and gibberellins (GAs) showed that there were major changes in the contents of these compounds associated with germination of after-ripened barley (Hordeum vulgare cv. Schooner and cv. Proctor) grain but not in hydrated dormant grain. Embryos from dormant and after ripened dry grain contained similar amounts of ABA, of ent-kaurenoids and of GAs, determined by gas chromatography-mass spectrometry-selected ion monitoring. In embryos of after-ripened grain, ABA content decreased rapidly after hydration and ABA appeared to be metabolized (inactivated) to phaseic acid (PA) rather than diffusing into the endosperm or the surrounding medium as previously thought. Similar changes in ABA occurred in hydrated dormant grain during germination in darkness. Accumulation of ent-kaurenoids and GAs, including GA1, the first biologically active GA in the early 13-hydroxylation biosynthetic pathway, occurred to a much greater extent in after-ripened than in dormant grain and these changes occurred mainly after 18 h of hydration when ABA had already decreased and germination was occurring. The block in ent-kaurenoid and GA synthesis in dormant grain appeared to occur prior to ent-kaurene in the biosynthetic pathway. These results are consistent with the view that ABA is the primary effector of dormancy and that after-ripening involves the development of the ability to reduce the amount of ABA quickly following hydration. Accumulation of GAs does not appear to be causally related to loss of dormancy but it does appear to be related to germination. PMID- 12081537 TI - Gibberellin regulates post-microsporogenesis processes in petunia anthers. AB - Previous studies have suggested that gibberellins (GAs) are produced in petunia anthers and transported to the corolla to induce growth and pigmentation. In this work, we studied the role of GA in the regulation of anther development. When petunia plants were treated with the GA-biosynthesis inhibitor paclobutrazol, anther development was arrested. Microscopic analysis of these anthers revealed that paclobutrazol inhibits post-meiotic developmental processes. The treated anthers contained pollen grains but the connective tissue and tapetum cells were degenerated. A similar phenotype was obtained when the Arabidopsis GA-signal repressor, SPY, was over-expressed in transgenic petunia plants, i.e. anther development was arrested following microsporogenesis. The expression of the GA induced gene, GIP, can be used in petunia as a molecular marker to study GA responses. GA3 treatment of young anthers promoted, and paclobutrazol inhibited, GIP expression, suggesting that the hormone controls the natural activation of the gene in the anthers. Analyses of GIP expression during anther development revealed that the gene is induced only after microsporogenesis. This observation further suggests a role for GA in the regulation of post-meiotic processes during petunia anther development. PMID- 12081538 TI - Far red end-of-day treatment restores wild type-like plant length in hybrid aspen overexpressing phytochrome A. AB - Shoot elongation in woody plants is modulated by a multitude of light signals, including irradiance, photoperiod and spectral composition, for which the phytochrome system is the probable photoreceptor. In hybrid aspen (Populus tremula x tremuloides) overexpression of the oat phytochrome A (PHYA) prevents growth cessation in response to short photoperiod, and plants exhibit dwarf growth that is related to reduced cell numbers and reduced gibberellin contents. End-of-day far-red treatment significantly enhances internode elongation in PHYA overexpressors as well as in the wild type, and this was found here to be caused by stimulation of cell division and cell extension. In PHYA overexpressors the effects were substantially larger than in the wild type, and resulted in complete restoration of wild type-like plant length as well as cell numbers, and gibberellin content was greatly increased. No clear effect of far-red end-of-day treatment on gibberellin levels could be detected in the wild type. It thus appears that the far-red end-of-day treatment might modify the responsiveness of the tissue to GA rather than the GA levels. The observed effects were completely reversed by a subsequent irradiation with red light. The present data show that dwarfism due to PHYA overexpression can be completely overcome by far red end-of day treatment, and the observations indicate that effects of far red end-of-day treatments appear to be mediated by phytochrome(s) other than phytochrome A. PMID- 12081539 TI - Pea rms6 mutants exhibit increased basal branching. AB - Our studies on two branching mutants of pea (Pisum sativum L.) have identified a further Ramosus locus, Rms6, with two recessive or partially recessive mutant alleles: rms6-1 (type line S2-271) and rms6-2 (type line K586). Mutants rms6-1 and rms6-2 were derived from dwarf and tall cultivars, Solara and Torsdag, respectively. The rms6 mutants are characterized by increased branching from basal nodes. In contrast, mutants rms1 through rms5 have increased branching from both basal and aerial (upper stem) nodes. Buds at the cotyledonary node of wild type (WT) plants remain dormant but in rms6 plants these buds were usually released from dormancy. Their growth was either subsequently inhibited, sometimes even prior to emergence above ground, or they grew into secondary stems. The mutant phenotype was strongest for rms6-1 on the dwarf background. Although rms6 2 had a weak single-mutant phenotype, the rms3-1 rms6-2 double mutant showed clear transgression and an additive branching phenotype, with a total lateral length almost 2-fold greater than rms3-1 and nearly 5-fold greater than rms6-2. Grafting studies between WT and rms6-1 plants demonstrated the primary action of Rms6 may be confined to the shoot. Young WT and rms6-1 shoots had similar auxin levels, and decapitated plants had a similar magnitude of response to applied auxin. Abscisic acid levels were elevated 2-fold at node 2 of young rms6-1 plants. The Rms6 locus mapped to the R to Gp segment of linkage group V (chromosome 3). The rms6 mutants will be useful for basic research and also have possible agronomical value. PMID- 12081540 TI - Photoinhibition and loss of photosystem II reaction centre proteins during senescence of soybean leaves. Enhancement of photoinhibition by the 'stay-green' mutation cytG. AB - The 'stay-green' mutation cytG in soybean (Glycine max) partially inhibits the degradation of the light-harvesting complex II (LHCII) and the associated chlorophyll during monocarpic senescence. cytG did not alter the breakdown of the cytochrome b6/f complex, thylakoid ATP synthase or components of Photosystem I. In contrast, cytG accelerated the loss of oxygen evolution activity and PSII reaction-centre proteins. These data suggest that LHCII and other thylakoid components are degraded by separate pathways. In leaves induced to senesce by darkness, cytG inhibited the breakdown of LHCII and chlorophyll, but it did not enhance the loss of PSII-core components, indicating that the accelerated degradation of PSII reaction centre proteins in cytG was light dependent. Illumination of mature and senescent leaves of wild-type soybean in the presence of an inhibitor (lincomycin) of chloroplast protein synthesis revealed that senescence per se did not affect the rate of photoinhibition in leaves. Likewise, mature leaves of the cytG mutant did not show more photoinhibition than wild-type leaves. However, in senescent cytG leaves, photoinhibition proceeded more rapidly than in the wild-type. We conclude that the cytG mutation enhances photoinhibition in senescing leaves, and photoinhibition causes the rapid loss of PSII reaction-centre proteins during senescence in cytG. PMID- 12081541 TI - Prevalence of Escherichia coli O157:H7 in industrial minced beef. AB - AIMS: The lack of baseline data on the prevalence of Escherichia coli O157:H7 in retail minced beef in France prompted this survey of industrial minced beef production. METHODS AND RESULTS: An automated enzyme-linked fluorescence immunoassay (ELFA), the VIDAS E. coli O157 method, was used to detect E. coli O157 in industrial minced beef samples. Confirmation of samples positive according to the ELFA was performed using an automated immunoconcentration (ICE) system, VIDAS ICE, which allows the selective capture and release of target organisms. The ICE was followed by culture on cefixime tellurite sorbitol MacConkey agar and a chromogenic medium, O157:H7 ID. Of the 3450 minced beef samples tested, 175 samples were positive with the ELFA method and, of these, four were confirmed by the ICE method. They were identified as sorbitol-negative, O157-positive, H7-positive, mobile, verotoxin-producing E. coli. CONCLUSIONS: The prevalence of E. coli O157:H7 in industrial French minced beef was 0.12%, consistent with many other reports. SIGNIFICANCE AND IMPACT OF THE STUDY: The low infective dose of E. coli O157:H7 presents a major threat. The main means of combating this organism are thermal destruction and good food hygiene covering activities on-farm, in the abattoir and in minced beef industries. PMID- 12081542 TI - In vitro studies on the effects of herbicides on the growth of rhizobia. AB - AIMS: To study the possible adverse effect of herbicides on nodulation and nitrogen fixation in legumes by affecting the nitrogen-fixing rhizobia. METHODS AND RESULTS: Experiments were conducted to study the effect of four herbicides (terbutryn/terbuthylazine, trietazine/simazine, prometryn and bentazone) on the growth of nitrogen-fixing pea rhizobia (Rhizobium leguminosarum) in vitro by measuring optical density. Terbutryn/terbuthylazine, trietazine/simazine and prometryn had an adverse effect on the growth of rhizobia whereas bentazone was safe to rhizobia. CONCLUSIONS: The above herbicides could be used in pea at the recommended rates. SIGNIFICANCE AND IMPACT OF THE STUDY: The adverse effects of herbicides on rhizobia were observed at concentrations not normally expected to occur under field conditions. Further, previously observed adverse effects of these herbicides on nodulation and nitrogen fixation of peas were, possibly, not due to their effects in rhizobia but to their adverse effects on the plant growth itself. PMID- 12081543 TI - Fungal chitosan production and its characterization. AB - AIMS: The objective of this investigation was to evaluate the chitosans produced by several species of fungi. METHODS AND RESULTS: Representatives of four species of filamentous fungi, Aspergillus niger, Rhizopus oryzae, Lentinus edodes and Pleurotus sajo-caju, and two yeast strains, Zygosaccharomyces rouxii TISTR5058 and Candida albicans TISTR5239, were investigated for their ability to produce chitosan in complex media. Fungal chitosan was produced at 10-140 mg g-1 cell dry weight, had a degree of deacetylation of 84-90% and a molecular weight of 2.7 x 104-1.9 x 105 Da with a viscosity of 3.1-6.2 centipoises (cP). CONCLUSIONS: Rhizopus oryzae TISTR3189 was found to be the producer of the highest amounts of chitosan. SIGNIFICANCE AND IMPACT OF THE STUDY: Commercial chitosan could be obtained from Rhizopus mycelia and would have potential applications for medical and agricultural uses. PMID- 12081544 TI - A study of antifungal antibiotic production by Streptomyces chattanoogensis MTCC 3423 using full factorial design. AB - AIMS: To determine the correct levels of the three process parameters identified as the most important for maximum production of antifungal antibiotic by the newly isolated strain of Streptomyces chattanoogensis MTCC 3423. METHODS AND RESULTS: The three independent variables, namely concentration of carbon source (glucose), nitrogen source (soybean meal) and temperature of incubation, were found to be the most important for production of antifungal antibiotic by the isolate Streptomyces chattanoogensis MTCC 3423 from a one-factor-at-a-time study. These variables were varied at three levels in 27 experiments using full factorial design. The results were analysed using statistical software for social sciences, SPSS (version 6.0). They indicated that the optimum combination of the three factors for maximum yield (263.63 U) of the antibiotic was concentration of carbon source (glucose) 5%, concentration of nitrogen source (soybean meal) 1% and temperature of incubation 30 degrees C. The antifungal antibiotic activity of the isolate was estimated against Candida albicans by bioassay. CONCLUSIONS: The S. chattanoogensis MTCC 3423 isolate was found to yield a maximum of 263.63 IU Nystatin ml-1 medium by manipulating the three process parameters of glucose, soybean meal and temperature of incubation. SIGNIFICANCE AND IMPACT OF THE STUDY: As process optimization by single-dimensional search is laborious and time consuming, and does not guarantee determination of optimal conditions, an attempt was made to use a more practical full factorial method. Even though antifungal antibiotic treatment and resistance data have been analysed using SPSS software, there are no reports on antifungal antibiotic production using SPSS. PMID- 12081545 TI - Respective effects of sodium and chloride ions on filament formation and growth and ethanol production in Zymomonas mobilis fermentations. AB - AIMS: To elucidate the separate effects of the cation and anion in Zymomonas mobilis cultures inhibited by NaCl. METHODS AND RESULTS: In batch cultures containing 0.175 mol l-1 NaCl, Z. mobilis developed filaments and growth and ethanol production were inhibited. Chloride ions (added as 0.175 mol l-1 NH4Cl) produced similar filamentous growth, while sodium ions at the same concentration (as Na2SO4) did not. Growth and ethanol production were more strongly inhibited by Na2SO4 than by NH4Cl. Ammonium and sulphate ions had no inhibitory effects at these concentrations. CONCLUSIONS: Filament formation in NaCl-inhibited Z. mobilis is due entirely to the chloride ion, while the sodium ion has the major effect on growth and ethanol production. SIGNIFICANCE AND IMPACT OF THE STUDY: To avoid filamentous growth, chloride ions should be excluded from ethanol fermentations employing Z. mobilis, e.g. sulphuric is preferable to hydrochloric acid when hydrolysing lignocellulosic substrates prior to fermentation. PMID- 12081546 TI - Typing of human and bovine Staphylococcus aureus by RAPD-PCR and ribotyping-PCR. AB - AIMS: To investigate genetic diversity among Staphylococcus aureus and to delineate the geographical distribution of the strains found. METHODS AND RESULTS: RAPD-PCR and ribotyping-PCR were employed for the characterization of Staph. aureus isolates from bovine and nosocomial origin. Among the strains, five to nine groups were distinguished by RAPD-PCR, depending on which primer was used, while ribotyping-PCR distinguished seven ribotypes. CONCLUSIONS, AND SIGNIFICANCE AND IMPACT OF THE STUDY: These results demonstrate the genetic heterogeneity of the strains studied, and the large dissemination of some clones throughout different regions and hosts, findings that may allow the monitoring of Staph. aureus infections. PMID- 12081547 TI - Influence of octanoic acid addition to medium on some volatile compounds and PR toxin biosynthesis by Penicillium roqueforti. AB - AIMS: The effect of wheat kernel medium supplementation with octanoic acid on the formation of PR toxin and some volatiles by Penicillium roqueforti was investigated. METHODS AND RESULTS: Octanoic acid was added to the medium once, prior to inoculation (4.55 mg g-1), or periodically (total 27.3 mg g-1) during the 10 day course of incubation. No octanoic acid was added to the reference sample. Levels of 2-heptanone, 2-heptanol and aristolochene, a volatile intermediate in PR toxin biosynthesis, were monitored using a solid phase microextraction (SPME) technique. The contents of PR toxin and ergosterol were determined after incubation. Aristolochene was observed in the reference sample, and 10.4 mg kg-1 of PR toxin was detected after 10 days. In cultures periodically supplemented with octanoic acid, no aristolochene or PR toxin were observed. However, in samples supplemented with octanoic acid only prior to incubation, the aristolochene level was 25% that in the reference sample, and PR toxin content was 3.4 mg kg-1. SIGNIFICANCE AND IMPACT OF THE STUDY: These data suggest that a high level of octanoic acid in the medium prevents PR toxin formation by P. roqueforti. PMID- 12081548 TI - Determination of Helicobacter pylori vacA allelic types by single-step multiplex PCR. AB - AIMS: To develop and evaluate a novel multiplex PCR assay that enables definition of Helicobacter pylori vacA allelic type in a single reaction. METHODS AND RESULTS: Application of the one-step system to DNA extracts from 22 cultures of known vacA genotype demonstrated that it was highly accurate. Analysis of 15 matched gastric biopsy/culture pairs generated exactly correlating genotype profiles. vacA genotypes were determined from an additional 62/70 gastric biopsies from dyspeptic patients of known H. pylori positive status by the one step assay, compared with 63/70 by the original two-reaction test. Types s1/m1, s1/m2 and s2/m2 were identified in 51.9%, 31.2% and 16.9% of biopsies, respectively. CONCLUSIONS: The multiplex PCR system developed enables rapid one step vacA genotyping that is accurate, easy to interpret and more economical than the alternative multiple-reaction tests. Application of this system to gastric biopsies from patients in South-east England demonstrated that s1/m1 was the most common genotype, while s1/m2 and s2/m2 were less prevalent. SIGNIFICANCE AND IMPACT OF THE STUDY: This simple one-step system can be applied direct to antral gastric biopsies without the need for culture, thereby facilitating rapid surveillance of vacA genotype in relation to geographical location and disease status. PMID- 12081549 TI - Biological activities of Saccharomyces cerevisiae and fermented rice bran as feed additives. AB - AIMS: The objective of the present study was to examine for the various biological activities for feed additives of Saccharomyces cerevisiae and fermented rice bran. METHODS: The feed additives (100 g of each: YE, brewery yeast; PM, mixture of YE and SF; SF, fermented rice bran) were decocted with 2 l water at 100 degrees C to a half volume. RESULTS: The hot-water extract of PM was found to increase the stimulation of the macrophage activation (2.0-fold) and the intestinal immune modulating activity (1.8-fold), compared with those of control. When the hot-water extracts of PM, SF and YE were fed to the rats for 8 d (1 g kg 1 d-1), PM and SF showed the more effective anti-stress effect on spleen, thyroid and thymus gland. SIGNIFICANCE AND IMPACT OF THE STUDY: The hot-water extract of PM can have a significant impact on strengthening the living body's immune system and proliferating anti-stress activities with relation to enhance immunity. PMID- 12081550 TI - Evaluation of the diversity of Paenibacillus polymyxa strains by using the DNA of bacteriophage IPy1 as a probe in hybridization experiments. AB - AIMS: To evaluate the genetic diversity within the species Paenibacillus polymyxa. METHODS: Southern hybridization was performed on 102 strains of P. polymyxa using DNA from the phage IPy1 as a probe. RESULTS: All 102 strains hybridized to phage IPy1 DNA. Data from different hybridization patterns obtained were used to construct a dendrogram in which 53 genotypic groups were split into two main clusters. One cluster contained strains from the rhizospheres of sorghum and maize planted in Cerrado soil, Brazil, and the majority of strains received from two culture collections. The other cluster contained strains isolated from different Brazilian soils and rhizospheres and strains deposited in a third culture collection. SIGNIFICANCE AND IMPACT OF THE STUDY: The approach used in this study appears to be a new and a very useful tool to study the diversity within this species. PMID- 12081552 TI - Comparison of arbitrarily primed-polymerase chain reaction and pulse-field gel electrophoresis for characterizing methicillin-resistant Staphylococcus aureus. AB - AIMS: The aim of this study was to analyse genotypes for clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA), including hetero-vancomycin resistant Staph. aureus (VRSA), at a Japanese university hospital. METHODS AND RESULTS: Seventy-eight clinical isolates of MRSA were analysed by arbitrarily primed-polymerase chain reaction (AP-PCR) using ERIC2 primer and by pulse-field gel electrophoresis (PFGE) following SmaI digestion. Analyses of the nine genotypes and 28 subtypes defined by PFGE, and of the three genotypes and 22 subtypes defined by AP-PCR, both facilitated epidemiological tracing. Used in combination, AP-PCR and PFGE provided more precise classification than the use of a single genotyping method. The six hetero-VRSA isolates were classified into four genotypes defined by the combination of both methods, but these genotypes contained non-VRSA isolates. CONCLUSIONS: The results suggest that both PFGE and AP-PCR are useful in discriminating MRSA, but not hetero-VRSA, isolates for epidemiological analysis. SIGNIFICANCE AND IMPACT OF THE STUDY: Combining the results of PFGE with the results of AP-PCR can provide more detail differentiation of MRSA and hetero-MRSA isolates than either method alone. PMID- 12081551 TI - Identification of Lactobacillus crispatus by polymerase chain reaction targeting S-layer protein gene. AB - AIMS: This study aimed to develop a polymerase chain reaction (PCR) method to identify Lactobacillus crispatus. METHODS AND RESULTS: A primer set (CbsA2F CbsA2R) for amplifying conserved regions of S-layer genes was designed to identify Lact. crispatus and the specificity of this set was compared with that of another primer set (Cri 16SI-Cri 16SII) which has been reported as a species specific primer set targeting the 16S rRNA gene. Among species in the Lact. acidophilus A1-A4 groups, when KOD polymerase was used for amplification, the primer set CbsA2F-CbsA2R gave PCR products with Lact. crispatus strains only. However, when Taq polymerase was used, this primer set gave products with one Lact. amylovorus strain as well as with Lact. crispatus strains. The primer set Cri 16SI-Cri 16SII gave PCR products with Lact. crispatus strains and two Lact. acidophilus strains, regardless of whether the polymerase used was KOD or Taq. CONCLUSIONS: A PCR targeting the S-layer gene and amplified with KOD polymerase can identify Lact. crispatus accurately and rapidly. SIGNIFICANCE AND IMPACT OF THE STUDY: To the authors' knowledge, this is the first paper to provide a PCR method for the specific identification of Lact. crispatus. PMID- 12081553 TI - Identification and characterization of an oligopeptide transport system in Leuconostoc mesenteroides subsp. mesenteroides CNRZ 1463. AB - AIMS: To identify and characterize an oligopeptide transport system in Leuconostoc mesenteroides CNRZ 1473. METHODS AND RESULTS: The uptake of a model substrate was monitored by determining intracellular concentrations of the corresponding amino acids by means of reversed-phase HPLC analysis. The oligopeptide transport system is specific for peptides containing at least four amino acid residues and operative under physiological conditions of growth. It is expressed maximally in the presence of oligopeptides, enhanced in the presence of Mg2+ or Ca2+ ions, and driven by ATP or a related energy-rich phosphorylated intermediate. CONCLUSIONS: The study showed evidence for and characterized the oligopeptide transport system of Leuc. mesenteroides for the first time. SIGNIFICANCE AND IMPACT OF THE STUDY: The potential of the findings is discussed with reference to the growth of Leuc. mesenteroides in mixed-strain cultures for the dairy industry. PMID- 12081554 TI - Conidium production by insect pathogenic fungi on commercially available agars. AB - AIMS: Conidium production by three species of insect pathogenic fungi, Metarhizium anisopliae, Beauveria bassiana and Verticillium lecanii, was assessed on various depths and types of commercially available agars. METHODS: Conidium production was assessed after 14 d of growth on commercially available media as well as at three different agar depths. RESULTS: Metarhizium anisopliae and B. bassiana isolates showed greatest conidium production on potato dextrose agar (PDA) at a depth of 2 mm, whereas V. lecanii showed greatest conidium production on yeast extract-peptone-dextrose agar (YPDA) regardless of agar depth. Optimum conidium production for M. anisopliae and B. bassiana was not only dependent upon the isolate used but also on the medium type and agar depth. SIGNIFICANCE AND IMPACT OF THE STUDY: Conidia are the infective structures for insect pathogenic fungi and this study suggests a rationale basis for consistent conidium production for laboratory and commercial practices. PMID- 12081555 TI - Characterization of a salt-tolerant extracellular a-amylase from Bacillus dipsosauri. AB - AIMS: The aims of this study were to purify and characterize an extracellular alpha-amylase from the salt-tolerant bacterium Bacillus dipsosauri. METHODS AND RESULTS: An extracellular alpha-amylase from B. dipsosauri strain DD1 was studied using the synthetic substrate 2-chloro-4-nitrophenyl-alpha-D-maltotrioside. Formation of the enzyme was induced by starch, repressed by D-glucose and highest after growth in medium containing 1.0 mol l-1 KCl. The alpha-amylase activity increased with KCl concentration, showed a pH optimum of 6.5, was stable up to 60 degrees C and was stimulated by 1.0 mol l-1 Na2SO4. The enzyme was purified from spent culture medium to apparent homogeneity by precipitation with ethanol, ion exchange chromatography on DEAE-cellulose, centrifugal membrane filtration and gel-filtration chromatography on BioGel P-100. The purified enzyme had a denatured molecular mass of about 80 kDa but behaved on non-denaturing polyacrylamide gels as if it had a mass of about 30 kDa. The enzyme was partially inhibited by glucose-containing oligosaccharides of increasing length and strongly inhibited by the divalent cations Cd2+ and Zn2+. CONCLUSIONS: The extracellular alpha-amylase from B. dipsosauri strain DD1 was purified to homogeneity and found to exhibit an unusually high degree of salt tolerance. SIGNIFICANCE AND IMPACT OF THE STUDY: The alpha-amylase from B. dipsosauri differs from previously described enzymes and may be useful for the processing of starches under high-salt conditions. PMID- 12081556 TI - Occurrence and strain diversity of thermophilic campylobacters in cattle of different age groups in dairy herds. AB - AIMS: To investigate the occurrence and numbers of thermophilic campylobacters excreted by cattle in dairy herds, and to assess the strain diversity within herds. METHODS AND RESULTS: Faecal samples from 15 animals at each of 24 cattle farms were cultured quantitatively for thermophilic campylobacters and 23% of animals and 83% of farms were positive for Campylobacter jejuni. Young animals had a higher prevalence and higher faecal concentration than older animals. Serotyping and pulsed field gel electrophoresis (PFGE) of isolates showed that the most common serotypes were 2, 4-complex and 11. Serotype 2 was especially prevalent among calves (68% of the positive calves). In eight of the 20 positive herds, all isolates had the same sero- and PFGE type while, in the other herds, two to five different types were isolated. CONCLUSIONS: Significant differences were found between age groups in relation to the prevalence and numbers of excreted campylobacters, serotype distribution and strain diversity. The relatively few different strains in each herd indicate that transmission between animals is common. SIGNIFICANCE AND IMPACT OF THE STUDY: The high prevalence on cattle farms of the human pathogen C. jejuni and the wide distribution of serotype 2, the most common serotype among Danish patients, indicate that cattle might be an important reservoir for human infections. The ability of this serotype to colonize calves in high numbers further indicates that serotype 2 strains may have an advantage over other serotypes. PMID- 12081557 TI - Pulsed high voltage electric discharge disinfection of microbially contaminated liquids. AB - AIMS: To examine the use of a novel multielectrode slipping surface discharge (SSD) treatment system, capable of pulsed plasma discharge directly in water, in killing micro-organisms. METHODS AND RESULTS: Potable water containing Escherichia coli and somatic coliphages was treated with pulsed electric discharges generated by the SSD. The SSD system was highly efficient in the microbial disinfection of water with a low energy utilization (eta approximately 10-4 kW h l-1). CONCLUSIONS: The SSD treatment was effective in the destruction of E. coli and its coliphages through the generation of u.v. radiation, ozone and free radicals. SIGNIFICANCE AND IMPACT OF THE STUDY: The non-thermal treatment method can be used for the eradication of micro-organisms in a range of contaminated liquids, including milk, negating the use of pasteurization. The method utilizes multipoint electric discharges capable of treating large volumes of liquid under static and flowing regimes. PMID- 12081558 TI - Hyperkalemia: An adaptive response in chronic renal insufficiency. AB - BACKGROUND: Hyperkalemia is a common feature of chronic renal insufficiency, usually ascribed to impaired K+ homeostasis. However, various experimental observations suggest that the increase in extracellular [K+] actually functions in a homeostatic fashion, directly stimulating renal K+ excretion through an effect that is independent of, and additive to, aldosterone. METHODS: We have reviewed relevant studies in experimental animals and in human subjects that have examined the regulation of K+ excretion and its relation to plasma [K+]. RESULTS: Studies indicate that (1) extracellular [K+] in patients with renal insufficiency correlates directly with intracellular K+ content, and (2) hyperkalemia directly promotes K+ secretion in the principal cells of the collecting duct by increasing apical and basolateral membrane conductances. The effect of hyperkalemia differs from that of aldosterone in that K+ conductances are increased as the primary event. The changes in principal cell function and structure induced by hyperkalemia are indistinguishable from the effects seen in adaptation to a high K+ diet. CONCLUSIONS: We propose that hyperkalemia plays a pivotal role in K+ homeostasis in renal insufficiency by stimulating K+ excretion. In patients with chronic renal insufficiency, a new steady state develops in which extracellular [K+] rises to the level needed to stimulate K+ excretion so that it again matches intake. When this new steady state is achieved, plasma [K+] remains stable unless dietary intake increases, glomerular filtration rate falls, or drugs are given that disrupt the new balance. PMID- 12081559 TI - Band 3 mutations, distal renal tubular acidosis, and Southeast Asian ovalocytosis. AB - Familial distal renal tubular acidosis (dRTA) and Southeast Asian ovalocytosis (SAO) may coexist in the same patient. Both can originate in mutations of the anion-exchanger 1 gene (AE1), which codes for band 3, the bicarbonate/chloride exchanger in both the red cell membrane and the basolateral membrane of the collecting tubule alpha-intercalated cell. Dominant dRTA is usually due to a mutation of the AE1 gene, which does not alter red cell morphology. SAO is caused by an AE1 mutation that leads to a nine amino acid deletion of red cell band 3, but by itself does not cause dRTA. Recent gene studies have shown that AE1 mutations are responsible for autosomal recessive dRTA in several countries in Southeast Asia; these patients may be homozygous for the mutation or be compound heterozygotes of two different AE1 mutations, one of which is usually the SAO mutation. PMID- 12081560 TI - Cyclosporine withdrawal and mycophenolate mofetil treatment effects on the progression of chronic cyclosporine nephrotoxicity. AB - BACKGROUND: Recent clinical trials of mycophenolate mofetil (MMF) in chronic allograft nephropathy (CAN) demonstrated that the dose of cyclosporine A (CsA) is one of the critical factors in determining graft function in CAN, but the effect of MMF on chronic CsA nephropathy is undetermined. We undertook this study to evaluate the effect of MMF on CsA-induced nephrotoxicity in an animal model of chronic CsA nephropathy. METHODS: In the first experiment, Sprague-Dawley rats on a low-salt diet were treated with CsA (7.5 mg/kg per day) for 10 weeks, or were treated with CsA for five weeks and then MMF (20 mg/kg per day) was administered five weeks later. In the second experiment, rats were treated with CsA for five weeks, and CsA was then withdrawn for five weeks with or without MMF treatment. Renal function, histologic parameters (tubulointerstitial fibrosis, arteriolopathy, ED-1-positive cells, renin-positive glomeruli, TUNEL-positive cells) and the expression of osteopontin and transforming growth factor (TGF) beta1 mRNA expressions were compared for different treatment groups. RESULTS: CsA treated rats showed decreased renal function and increased histologic parameters compared with the vehicle (VH)-treated rats. The addition of MMF did not improve these parameters compared with the CsA-treated rats. With CsA withdrawal, renal function and histologic parameters were significantly improved compared with the CsA-treated rats, and MMF treatment after CsA withdrawal further improved the histologic parameters. At the molecular level, the addition of MMF did not decrease the expression of osteopontin and transforming growth factor-beta1 (TGF beta1) mRNAs, which were increased in the CsA-treated rat kidney. With CsA withdrawal, the expression of both mRNAs was significantly decreased compared with the CsA group, and a further decrease was observed with MMF treatment after CsA was withdrawn. CONCLUSION: The combined treatment of CsA and MMF does not prevent the development of chronic CsA nephrotoxicity, but MMF treatment after CsA withdrawal does improve chronic CsA nephrotoxicity. This finding provides a rationale for MMF treatment in chronic CsA nephrotoxicity. PMID- 12081561 TI - Interleukin-1beta induces human proximal tubule cell injury, alpha-smooth muscle actin expression and fibronectin production. AB - BACKGROUND: Tubulointerstitial lesions, characterized by tubular injury, interstitial fibrosis and the appearance of myofibroblasts, are the strongest predictors of the degree and progression of chronic renal failure. These lesions are typically preceded by macrophage infiltration of the tubulointerstitium, raising the possibility that these inflammatory cells promote progressive renal disease through fibrogenic actions on resident tubulointerstitial cells. The aim of the present study, therefore, was to investigate the potentially fibrogenic mechanisms of interleukin-1beta (IL-1beta), a macrophage-derived pro-inflammatory cytokine, on human proximal tubule cells (PTC). METHODS: Confluent, quiescent, passage 2 PTC were established in primary culture from histologically normal segments of human renal cortex (N = 11) and then incubated in serum- and hormone free media supplemented with either IL-1beta (0 to 4 ng/mL) or vehicle (control). RESULTS: IL-1beta significantly enhanced fibronectin secretion by up to fourfold in a time- and concentration-dependent fashion. This was accompanied by significant (2.5- to 6-fold) increases in alpha-smooth muscle actin (alpha-SMA) expression, transforming growth factor beta (TGF-beta1) secretion, nitric oxide (NO) production, NO synthase 2 (NOS2) mRNA and lactate dehydrogenase (LDH) release. Cell proliferation was dose-dependently suppressed by IL-1beta. NG methyl-l-arginine (L-NMMA; 1 mmol/L), a specific inhibitor of NOS, blocked NO production but did not alter basal or IL-1beta-stimulated fibronectin secretion. In contrast, a pan-specific TGF-beta neutralizing antibody significantly blocked the effects of IL-1beta on PTC fibronectin secretion (IL-1beta, 268.1 +/- 30.6 vs. IL-1beta+alphaTGF-beta 157.9 +/- 14.4%, of control values, P < 0.001) and DNA synthesis (IL-1beta 81.0 +/- 6.7% vs. IL-1beta+alphaTGF-beta 93.4 +/- 2.1%, of control values, P < 0.01). CONCLUSION: IL-1beta acts on human PTC to suppress cell proliferation, enhance fibronectin production and promote alpha-smooth muscle actin expression. These actions appear to be mediated by a TGF-beta1 dependent mechanism and are independent of nitric oxide release. PMID- 12081562 TI - Role of c-SRC and ERK in acid-induced activation of NHE3. AB - BACKGROUND: In the renal proximal tubule, chronic acidosis causes increases in apical membrane NHE3 activity, which serve to increase transepithelial H+ secretion and return systemic pH to normal levels. Incubation of cultured renal epithelial cells in acid media activates c-Src. METHODS: OKP cells were incubated in control (pH 7.4) or acid (7.0) media, and NHE3 activity measured as cytoplasmic pH (pHi) recovery from an acid load using BCECF. c-Src, ERK, and JNK kinase activities were measured by immune complex kinase assays with enolase, MBP, and GST-c-Jun, respectively, as substrates in the in vitro assays. To determine the role of c-Src in acid-induced NHE3 activation, cells were transfected with vector alone or a dominant negative c-Src (c-SrcK295M). RESULTS: Expression of dominant negative c-srcK295M in OKP cells prevented acid-induced activation of NHE3. Incubation of OKP cells in acid media increased ERK activity and c-fos expression, but did not increase JNK activity. Acidosis in vivo also activated renal cortical c-Src and ERK kinases, whereas incubation of 3T3 cells in acid media activated c-Src but not ERK kinase. Expression of c-srcK295M did not affect ERK or c-fos activation by acid incubation. Inhibition of MEK with PD98059 inhibited activation of NHE3 by acid incubation. CONCLUSIONS: These studies suggest that acidosis activates c-Src and MEK/ERK/c-fos. While both pathways are necessary for activation of NHE3, they are activated independently. PMID- 12081563 TI - Intraperitoneal protein injection in the axolotl: the amphibian kidney as a novel model to study tubulointerstitial activation. AB - BACKGROUND: A substantial body of experimental evidence suggests that protein loading causes activation of proximal tubular epithelial cells with consecutive interstitial fibrosis. These studies have mostly been performed using mammalian in vivo models of glomerular damage or tissue cultures of mammalian tubulointerstitial cells. The kidney of the axolotl contains not only closed nephrons, but also nephrons with ciliated peritoneal funnels called nephrostomes that have access to the peritoneal fluid. Injection of protein into the peritoneal cavity fails to expose closed nephrons to a protein load, but causes selective uptake and transient storage of proteins in tubular epithelial cells of nephrons with nephrostomes. The purpose of the present study was to determine whether (a) the axolotl kidney can be used as a model to assess protein uptake by tubular cells in vivo in the absence of glomerular damage, and (b) this is accompanied by any evidence of tubular epithelial cell activation and interstitial fibrosis. METHODS: Male and female axolotl (80 to 120 g of weight) were given a daily intraperitoneal injection of 1.5 mL endotoxin-free calf serum or saline as control. Kidneys were harvested after 4 or 10 days using perfusion fixation for light microscopy (fibrous tissue stain) and saline perfusion for immunohistochemistry (fibronectin, TGF-beta and collagen I). RESULTS: The findings document selective storage of protein and lipids, progressive with time, in proximal tubular epithelial cells of nephrons draining the coelomic cavity. In addition, progressive focal accumulation of fibrous tissue was noted around protein-storing tubules. Immunohistochemical staining demonstrated the presence of fibronectin and TGF-beta in the tubular epithelial cells and interstitial cells. CONCLUSION: The axolotl kidney provides a novel in vivo model to study tubulointerstitial activation and induction of interstitial fibrosis by protein loading. The findings are independent of alterations of glomerular function that may have potential confounding effects on peritubular hemodynamics, pO2, cell traffic, etc. PMID- 12081564 TI - Inhibition of apoptosis in rat mesangial cells by tissue inhibitor of metalloproteinase-1. AB - BACKGROUND: Tissue inhibitor of metalloproteinase-1 (TIMP-1) is an important inhibitor of extracellular matrix degradation. Recently, it was reported that TIMP-1 also could inhibit apoptosis in B type lymphocyte. This study was designed to examine the effects of TIMP-1 on mesangial cell apoptosis. METHODS: The full length cDNA of TIMP-1 was cloned and used to construct two recombinant vectors, TIMP-1S and TIMP-1AS, encoding sense TIMP-1 and antisense TIMP-1, respectively. The vectors were transfected into rat mesangial cells (RMC) and their expressions detected by Northern and Western blotting. Apoptosis was induced by serum deprivation, and was monitored for DNA fragmentation by TUNEL assay and DNA laddering. In addition, the expression of endogenous TIMP-1, matrix metalloprotein-2 (MMP-2), and MMP-9, as well as apoptosis-related genes Bcl-2 and Bax were investigated. RESULTS: TIMP-1AS transfection induced a suppression of TIMP-1 expression accompanied by an earlier onset of apoptosis, and TIMP-1S transfection induced TIMP-1 over-expression accompanied by a much later onset of apoptosis. A neutralizing antibody of TIMP-1 restored the sensitivity of TIMP-1S transfected RMC to serum deprivation, but a synthetic matrix metalloproteinase inhibitor BB-94 did not influence the sensitivity of TIMP-1S-transfected RMC to serum deprivation. Finally, TIMP-1 over-expression inhibited the expression of Bax but with no effect on the expression of Bcl-2. CONCLUSION: TIMP-1 inhibits the serum deprivation-induced apoptosis in RMC, in which Bax might be involved. PMID- 12081565 TI - Cytoprotective effects of nitrates in a cellular model of hydronephrosis. AB - BACKGROUND: The earliest insult to the kidney following the onset of ureteral obstruction is a marked elevation in collecting system pressure. This imparts a mechanical stress that is transmitted directly from the collecting system to the kidney substance. Renal tubular injury is the principal functional and histological change encountered, with glomerular changes being less marked and occurring later. Nitric oxide (NO) has been shown to protect against renal injury in UO, but its mode of action has not been clearly defined. METHODS: MDCK (canine) and HK-2 (human) renal tubular cells were grown under control conditions or subjected to mechanical strain for periods of 24 and 48 hours. Cells were studied treated with or without Fas-antibody, etoposide or diethyl maleate (DEM) alone or in combination with NG-monomethyl l-arginine (L-NMMA), sodium nitroprusside (SNP) or l-arginine. Cell proliferation and apoptosis was determined using propidium iodide DNA staining. NO production and inducible NO synthase (iNOS) expression were measured by the Griess reaction and Western blotting, respectively. RESULTS: Cells subjected to mechanical strain displayed a decrease in the proportion of cells undergoing cell division. They also showed an increased susceptibility to apoptosis. Associated with this was a decrease in Bcl 2 expression. An increase in iNOS expression was seen in cells subjected to mechanical strain, but no increase in NO production. The cellular effects of mechanical strain were reversed by SNP and l-arginine. CONCLUSIONS: Culture of renal tubule cells in an environment of mechanical strain results in an imbalance in homeostasis and a net cell loss. This can be reversed by the administration of an NO donor or precursor. PMID- 12081567 TI - The flux of arginine after ischemia-reperfusion in the rat kidney. AB - BACKGROUND: Renal arginine synthesis is regulated by arginine plasma levels. The amino acid arginine is synthesized in the proximal tubule of the kidney. Renal ischemia reperfusion (I-R) injury as seen after shock, trauma and major vascular surgery, leading to acute tubular necrosis, might reduce arginine production. METHODS: Wistar rats received either bovine liver arginase (ASE), to lower arginine plasma levels, or saline (SAL). Following the ASE or SAL infusion, rats were randomized to receive a renal artery clamp for 70 minutes, followed by 150 minutes of reperfusion. Renal arteriovenous blood samples were measured and plasma flow was calculated in the I-R kidney (SAL/I-R and ASE/I-R) and the contralateral kidney (SAL/C-L and ASE/C-L) in order to determine renal arginine metabolism. RESULTS: Arginase infusion resulted in lower arginine plasma levels compared to SAL treatment (SAL/I-R vs. ASE/I-R, P < 0.005, and SAL/C-L vs. ASE/C L, P < 0.005). Renal plasma flow was similar for all groups. The kidney switched from arginine production into arginine uptake after ischemia reperfusion (SAL/I-R vs. SAL/C-L, P < 0.01, and ASE/I-R vs. ASE/C-L, P < 0.01). Renal uptake of glutamine and citrulline increased after ischemia reperfusion (SAL/I-R vs. SAL/C L and ASE/I-R vs. ASE/C-L, both P < 0.01). Histopathological slices of the kidney showed significantly higher counts of hyperchromasia, pyknosis, nuclear fragmentation and mitoses in individual kidney cells after ischemia reperfusion. CONCLUSION: Decreased renal arginine production is observed with unilateral ischemia-reperfusion, and this change in arginine flux could contribute to or slow the recovery from the low plasma levels of arginine seen in conditions like trauma, shock, or after vascular procedures. PMID- 12081566 TI - Effects of cyclosporine in osteopontin null mice. AB - BACKGROUND: Osteopontin (OPN) is a macrophage adhesive and cell survival factor that is up-regulated in tubules in tubulointerstitial disease. We have previously reported that rats with cyclosporine (CsA) nephropathy have increased tubular osteopontin that correlates with the infiltration of macrophages and interstitial fibrosis. This study tested the hypothesis that the absence of OPN would ameliorate CsA nephropathy. METHODS: OPN knockout (-/-) and wild type (+/+) mice were fed a low salt diet (Na+ 0.01%) for one week and then received daily CsA injections (30 mg/kg, SC) until sacrifice at two weeks. Afferent arteriolopathy, tubulointerstitial injury, macrophage infiltration, collagen III deposition, transforming growth factor-beta (TGF-beta) expression, and tubular and interstitial cell proliferation and apoptosis were evaluated. RESULTS: Wild type mice developed early features of CsA nephropathy, with arteriolar hyalinosis and cortical and tubulointerstitial fibrosis. Despite comparable CsA levels, OPN-/- mice had less arteriolopathy (15 vs. 24%, P < 0.05), a 20% reduction in cortical macrophage infiltration (P < 0.05), and 20% reduction in interstitial collagen deposition (P < 0.05). OPN-/- mice also showed less cortical interstitial cell proliferation but no differences in tubular cell proliferation or apoptosis. OPN+/+ mice also developed some neurotoxicity, consisting of ataxia, and this was associated with increased mortality at two weeks. CONCLUSION: OPN partially mediates arteriolopathy, early macrophage recruitment and fibrosis in murine CsA nephropathy. OPN also may be involved in CsA associated neurotoxicity. PMID- 12081568 TI - P-selectin-dependent macrophage migration into the tubulointerstitium in unilateral ureteral obstruction. AB - BACKGROUND: Interstitial infiltration of macrophages (Mo) is one of the main causal factors for the tubulointerstitial injury. However, precise mechanisms of Mo infiltration into tubulointerstitium have not been fully explored. The purposes of this study were to assess the role of selectins in the acute infiltration of Mo in rats with unilateral ureteral obstruction (UUO) and to evaluate the role of vasa recta, that is, whether they facilitate massive influx of Mo into the interstitium by functioning as specialized vessels. METHODS: To evaluate the role of selectins in Mo infiltration into tubulointerstitium, the expression of selectins and L-selectin ligands was examined by immunohistochemistry and immunoelectron microscopy. The functional role of P selectin in vasa recta was studied by Stamper-Woodruff assay, in vivo p-Mo migration assay and in vivo blocking experiments with the monoclonal antibody (mAb) ARP2-4. RESULTS: Selective expression of P-selectin was detected in vasa recta as early as one hour after UUO, and the expression increased thereafter for 96 hours. In contrast, endothelial expression of L-selectin ligands and E selectin were not detectable. In the Stamper-Woodruff assay on kidney sections of rats with UUO, the adhesion of isolated rat peritoneal Mo (p-Mo) to vasa recta was significantly inhibited by the mAb ARP2-4 (P-selectin blocker; P < 0.01), but not by mAb ARE-5 (E-selectin blocker) or rLECIg (rat L-selectin chimeric protein). In the in vivo transfer experiments with fluorescein-labeled p-Mo into rats 48 hours after UUO, labeled p-Mo had accumulated around vasa recta at three minutes and had infiltrated predominantly into the outer medulla at 180 minutes. The number of labeled p-Mo was reduced when the rats were pretreated with ARP2-4 (P < 0.01). Finally, ARP2-4 (10 mg/kg), injected 15 minutes before UUO, reduced the number of infiltrated Mo (P < 0.01). CONCLUSION: The results suggest that vasa recta, which express P-selectin, contribute to massive infiltration of Mo into the interstitium by functioning as specialized post-capillary venules. PMID- 12081569 TI - Cloning and expression of rat caspase-6 and its localization in renal ischemia/reperfusion injury. AB - BACKGROUND: Caspase-6 is an important member of the executioner caspases in the caspase family of cell death proteases. The executioner caspases are the major active caspases detected in apoptotic cells and are generally considered to mediate the execution of apoptosis by cleaving and inactivating intracellular proteins. However, the complete characterization of mRNA and protein of caspase-6 in rat and its expression in normal kidney and in disease state has not been previously elucidated. METHODS: A rat kidney cortex lambdagt10 cDNA library was screened to isolate the full-length caspase-6 cDNA. The recombinant caspase-6 protein was characterized by expression in bacteria and by transient transfection in mammalian cells. The expression in various tissues was analyzed by Northern blot, and localization in normal and ischemic kidney was performed by immunohistochemistry. RESULTS: The predicted amino acid sequence of rat caspase-6 contains 277 amino acids, with two potential glycosylation sites, an integrin binding site (KGD), the caspase active site pentapeptide QACRG and the caspase family signature, HX2-4(S,C) X4(L,I,V,M,F)2(S,T)HG (HVDADCFVCVFLSHG). Rat caspase 6 is unique among known caspases by possessing a relatively long 5' untranslational region. Among various tissues tested, cas-pase-6 was expressed in varying levels in kidney, liver, spleen, heart, muscle, testis, and lung. Bacterial expression of recombinant rat caspase-6 resulted in production of both of the pro-form and active form of the enzyme suggesting autoactivation. Transient overexpression of rat caspase-6 in COS-1 cells induced DNA fragmentation, a hallmark of apoptosis. We also examined the localization and expression of caspase-6 by immunohistochemistry in kidneys subjected to 40 minutes of ischemia followed by 24 hours of reperfusion injury. Normal kidney showed mostly cytoplasmic and some nuclear staining of the tubules. Kidneys 24 hours after 40 minutes of ischemia showed more intense and diffused cytoplasmic staining with prominent nuclear staining, indicating increased expression and translocation from the cytoplasm to the nuclei. The staining in glomeruli was negative in both normal and ischemic kidney. CONCLUSIONS: These studies demonstrate cloning, expression and characterization of the full-length rat caspase-6 and its localization in normal kidneys and kidneys subjected to ischemia/reperfusion injury. Since caspase-6 is involved in the degradation of nuclear matrix proteins and in activation of caspase-3, it may play an important role during renal ischemic injury. PMID- 12081570 TI - Antibody-induced albuminuria and accelerated focal glomerulosclerosis in the Thy 1.1 transgenic mouse. AB - BACKGROUND: Podocytes play an important role in the development of proteinuria and focal glomerulosclerosis. Previously we have demonstrated that a combination of two monoclonal antibodies (mAb) against aminopeptidase A (APA), an enzyme present on podocytes, induces a massive acute albuminuria in mice. The present study examined the relationship between the acute antibody-induced albuminuria and the development of focal glomerulosclerosis in the Thy-1.1 transgenic mouse. This mouse expresses a hybrid human-mouse Thy-1.1 antigen on the podocytes, and slowly but spontaneously develops albuminuria and focal glomerulosclerosis. METHODS: Five-week-old non-albuminuric Thy-1.1 transgenic and non-transgenic control mice were injected with anti-APA and anti-Thy-1.1 mAb or saline. Albuminuria was measured at days 1, 7, 14 and 21. At day 21 kidneys were processed for light microscopy, immunofluorescence, and electron microscopy. RESULTS: Injection of anti-APA and anti-Thy1.1 mAb in Thy-1.1 transgenic mice induced an albuminuria at day 1 that persisted at day 21. The acute albuminuria after injection of anti-APA mAb was more prominent but transient in non transgenic mice. In non-trangenic mice no albuminuria could be induced with anti Thy 1.1 mAb. Light microscopy revealed normal glomeruli at day 1 in all transgenic mice, however, at day 21 advanced glomerulosclerotic lesions were seen in mice injected with either anti-APA mAb (37+/-19% of glomeruli affected) or anti-Thy-1.1 mAb (71+/-5%). Non-transgenic mice did not reveal sclerotic lesions at any time investigated. In the transgenic mice the percentage of focal glomerulosclerosis at day 21 did not correlate with albuminuria at day 21. However, we found a highly significant correlation between percentage of focal glomerulosclerosis and the time-averaged albuminuria over the three-week study period (P < 0.001). CONCLUSION: Injection of a combination of anti-APA or anti Thy-1.1 mAb into one mo old, non-albuminuric Thy-1.1 transgenic mice induces an acute albuminuria at day 1 that is accompanied by an accelerated focal glomerulosclerosis at day 21. We suggest that the Thy-1.1 transgenic mouse is an excellent model to study specifically the relation between podocytic injury, albuminuria and the development of focal glomerulosclerosis. PMID- 12081571 TI - Studies of renal injury IV: The GLUT1 gene protects renal cells from cyclosporine A toxicity. AB - BACKGROUND: Renal cells activate the GLUT1 gene when exposed to stress. This response promotes glucose influx and glycolysis, which protects cells and preserves viability. We tested the hypothesis that cytotoxicity from cyclosporine A (CsA), a valuable but nephrotoxic immunosuppressor, also activated the GLUT1 gene. Methods and Results. GLUT1 nuclear transcription was increased in LLCPK1 cells injured with CsA, 10-5 mol/L or more for 24 hours, with increases of GLUT1 mRNA and protein levels, resulting in greater glucose consumption and glycolysis. The integrated stress response to CsA toxicity was cytoprotective, as blockade of glucose influx and glycolysis with 10-4 mol/L phloretin magnified CsA toxicity. Remarkably, whereas phloretin reduced GLUT1 transcription, it still increased GLUT1 protein and mRNA levels, and even amplified their responses to CsA. Interestingly, intracellular pH was preserved despite of greater lactic acid production in the face of Na+/H+ exchange inhibition from CsA toxicity. However, further inhibition of Na+/H+ exchange with amiloride greatly magnified CsA toxicity and GLUT1 gene transcription. CONCLUSION: Activation of the GLUT1 gene during renal cell injury is mediated by at least two redundant systems. CsA stimulates GLUT1 gene transcription when membrane transport delivers glucose to the cell. However, when glucose delivery is compromised, GLUT1 gene expression is still supported by alternative mechanisms that remain operational even after cellular energy metabolism is compromised further by inhibition of glucose and glycolytic fluxes. PMID- 12081572 TI - Epithelial-mesenchymal transition of tubular epithelial cells in human renal biopsies. AB - BACKGROUND: In recent studies performed on cultured cells and experimental nephropathies, it has been hypothesized that tubular epithelial cells (TEC), via epithelial-mesenchymal transformation (EMT), can become collagen-producing cells. According to this theory, they should proceed through several activating steps, such as proliferation and phenotype changes, to eventually synthesize extracellular matrix (ECM). METHODS: To evaluate whether EMT operates in human TECs, 133 renal biopsies of different renal diseases were studied, analyzing by immunohistochemistry and in situ hybridization the possible expression of markers of proliferation (PCNA, Mib-1), cellular phenotype (vimentin, alpha-SMA, cytokeratin, ZO-1) and ECM production (prolyl 4-hydroxylase, HSP47, interstitial collagens). RESULTS: Independently of histological diagnosis, variable degrees of TEC positivity for PCNA (2.7 +/- 2.4 cells/field) and Mib-1 (1.9 +/- 2.3) were present. TECs expressing vimentin (1.4 +/- 4.7) and alpha-smooth muscle actin (alpha-SMA; 0.04 +/- 0.4) also were detected. It was possible to observe loss of epithelial antigens from 8 to 10% of the tubular cross sections. Moreover, TECs were stained by prolyl 4-hydroxylase (3.6 +/- 4.3), heat shock protein-47 (HSP47; 2.9 +/- 5.4), collagen type I (0.2 +/- 2.7) and type III (0.3 +/- 2.0). Collagen types I and III mRNAs were found in 0.8 to 1.4 cells/field. The number of TEC with EMT features were associated with serum creatinine and the degree of interstitial damage (P< or = 0.03), and even considering the 45 cases with mild interstitial lesions, the tubular expression of all markers remained strictly associated with renal function (P< or = 0.01). CONCLUSIONS: Our results suggest that, via transition to a mesenchymal phenotype, TEC can produce ECM proteins in human disease and directly intervene in the fibrotic processes. Moreover, the association of EMT features with serum creatinine supports the value of these markers in the assessment of disease severity. PMID- 12081573 TI - Expression of the iron transporter DMT1 in kidney from normal and anemic mk mice. AB - BACKGROUND: DMT1 (Nramp2/DCT1) is the major apical iron transporter in absorptive cells of the duodenum, but also transports transferrin-iron across the membrane of acidified endosomes in peripheral tissues. DMT1 mRNA and protein expression has been detected in rat and mouse kidney, but its role at that site remains to be clarified. METHODS: Immunoblotting and immunohistochemistry with specific affinity purified anti-DMT1 polyclonal antibodies were used to study expression and localization of DMT1 in mouse kidney. Possible regulation of DMT1 protein expression by the body iron stores also was examined in normal mice deprived of dietary iron, and in the genetically anemic mk mice that bear a loss of function mutation at DMT1 (G185R). RESULTS: In microsomal kidney fractions, DMT1 isoform I (encoded by the iron responsive element (IRE)-containing mRNA) is detected as an abundant 70 to 75 kD membrane protein. DMT1 is expressed in the cortex and not in the medulla, and is present at the brush border and apical pole of epithelial cells of proximal tubules. In contrast to the intestine, DMT1 protein expression in kidney is only slightly increased upon deprivation of dietary iron, suggesting different regulation at the two sites. In kidneys from mk/mk mice, the level of detectable DMT1(G185R) protein is drastically decreased compared to mk/+ controls. CONCLUSION: These results suggest that DMT1 may act as a re-uptake system for divalent cations at the brush border of kidney proximal tubules. A pathological mutation at DMT1 affects targeting/expression of the protein in the kidney. PMID- 12081574 TI - Na+/H+ exchange activity and NHE-3 expression in renal tubules from the spontaneously hypertensive rat. AB - BACKGROUND: The NHE-3 isoform of the Na+/H+ antiporter, in the apical membrane of renal proximal tubule, is responsible for the bulk transport of Na+ and fluid reabsorption. Studies have reported that apical NHE-3 translocates to internal pools, thereby facilitating natriuresis when blood pressure increases abruptly. METHODS: The present study examined Na+/H+ exchange activity and NHE-3 expression in renal cortical tubules from the spontaneously hypertensive rat (SHR) and WKY rats before and after the development of hypertension. SHR 4 to 6 weeks of age were pre-hypertensive, 6 to 7 weeks old had mild hypertension, and 8 to 13 weeks old had severe hypertension. Renal proximal tubules (PTs) were isolated and purified by Percoll gradient centrifugation. NHE-3 protein and mRNA levels were determined by Western and Northern blots, respectively. Apical brush border membrane vesicles (BBMV) were prepared using the MgSO4 aggregation method and Na+/H+ exchange activity assessed using the acridine orange method. RESULTS: Na+/H+ exchange activity, determined as the rate of Na+-dependent intracellular pH (pHi) recovery assessed using BCECF after an acute acid load, was significantly greater in PTs from SHR than in WKY rats at all age groups (4 to 6 weeks, 0.30 +/- 0.04 vs. 0.24 +/- 0.02 pH U/30 sec, P < 0.05; 6 to 7 weeks, 0.42 +/- 0.07 vs. 0.29 +/- 0.05 pH U/30 sec, P < 0.05; and 8 to 13 weeks, 0.48 +/- 0.07 vs. 0.40 +/- 0.07 pH U/30 sec, P < 0.05). The Na+-dependent recovery in BBMV was also greater in SHR than WKY rats (1464 +/- 62 vs. 1042 +/- 79 fluorescence. U/5 sec, P < 0.001) and was unaffected by cariporide, a specific NHE-1 inhibitor. NHE-3 protein levels also were significantly higher in SHR than age-matched WKY rats at all stages during the development of hypertension (pre-hypertensive 1.8 fold; early onset hypertension twofold; established hypertension 1.5-fold; each P < 0.05). By contrast, NHE-3 mRNA levels were not different between SHR and WKY rats at each age group. CONCLUSIONS: Na+/H+ exchange activity and NHE-3 protein abundance in renal proximal tubules from the SHR are increased while NHE-3 mRNA is not. A post-transcriptional event(s) best explains the increase in NHE-3 protein expression since mRNA levels were not increased. The alterations in the SHR antedate the development of hypertension and fail to decrease as blood pressure increases with age in the SHR, which likely results in inappropriate renal sodium retention in the face of a chronic rise in blood pressure. PMID- 12081575 TI - Central aortic pressure augmentation in stable renal transplant recipients. AB - BACKGROUND: Premature cardiovascular disease is the leading cause of death in renal transplant recipients and classical risk factors significantly underestimate the risk. The increased effect of arterial wave reflections on central arteries has recently been shown to be an important independent predictor of cardiovascular mortality in chronic hemodialysis patients. The aim of this study was to assess the contribution of several classical and potential non classical cardiovascular risk factors on aortic pressure augmentation by the reflected arterial wave in stable renal transplant recipients. METHODS: Using the non-invasive technique of pulse wave analysis aortic augmentation was investigated in 250 stable renal transplant recipients. Peripheral pulse waveforms were recorded from the radial artery. Central aortic waveforms were then generated and the aortic augmentation index calculated. RESULTS: In multivariate analysis, female sex (regression coefficient 7.5 +/- 1.7%; P < 0.001), heart rate (-4.8 +/- 0.5% per 10 beats/min; P < 0.001), mean arterial pressure (4.2 +/- 0.6% per 10 mm Hg; P < 0.001), the persistence of an arteriovenous fistula (4.1 +/- 1.3%; P < 0.005), total time on renal replacement therapy (3.8 +/- 0.9% per 10 years; P < 0.001), height (-3.1 +/- 0.8% per 10 cm; P < 0.001), immunosuppression with cyclosporine (2.8 +/- 1.3%; P < 0.005) and age (2.5 +/- 0.5% per 10 years; P < 0.001) were all important correlates of aortic augmentation index. CONCLUSIONS: Our findings suggest, to our knowledge for the first time, that both the presence of a functioning arteriovenous fistula and immunosuppressive treatment with cyclosporine are associated with an increased aortic augmentation index in renal transplant recipients and could, therefore, be potential reversible contributors to the high cardiovascular risk profile in these patients. PMID- 12081576 TI - Hypertensive nephrosclerosis in African Americans versus Caucasians. AB - BACKGROUND: Extensive global glomerulosclerosis (GS) has been reported in African Americans with hypertension and renal insufficiency, far exceeding that in Caucasians. To assess and compare severity and phenotype of injury in biopsied African Americans and Caucasians who morphologically had hypertensive nephrosclerosis, we performed a retrospective biopsy study. METHODS: All renal biopsies with a histological diagnosis of hypertensive nephrosclerosis from the last 11 years were identified from our clinical files. Lesions of global and segmental sclerosis, interstitial fibrosis and vascular sclerosis were semiquantitatively analyzed as percent involved, or on a 0 to 3 scale, respectively. The phenotypes of global glomerulosclerosis also were categorized as either the solidified (that is, the entire tuft is solidified) or the obsolescent type (that is, Bowman's space is occupied by collagenous material and the tuft is retracted). RESULTS: Sixty-two patients (19 African Americans, 43 Caucasians) were included in the study. At biopsy, African Americans were younger than Caucasians with higher serum creatinine, but no difference in proteinuria or mean arterial pressure (MAP). African Americans had a marked increase in the solidified form of GS (25 +/- 6 in African Americans vs. 8 +/- 2% in Caucasians, P < 0.01). This extensive solidification of glomeruli was associated with segmental sclerosis in African Americans (38 +/- 10%), contrasting low prevalence of solidified GS in Caucasians with segmental sclerosis (10 +/- 3%, P < 0.05) and in African Americans without segmental sclerosis (10 +/- 4%, P < 0.05). African Americans with segmental sclerosis were younger and clinically expressed a more severe renal disease than Caucasians with this lesion. Interstitial fibrosis was greater in African Americans than in Caucasians (54 +/- 6 vs. 33 +/- 3%, P < 0.01) and correlated with proteinuria and serum creatinine levels, especially in African Americans, and also with GS. Vascular sclerosis was worse in African Americans than in Caucasians (0.96 +/- 0.04 vs. 0.77 +/- 0.08 score, P < 0.05) and did not correlate with GS. By modeling, neither MAP nor age was useful in predicting any morphological lesions and proteinuria accounted only minimally for the variability of GS. CONCLUSIONS: Blood pressure levels and proteinuria did not account for morphological lesions, suggesting other factors (such as genetic factors, microvascular disease) may play a role. The phenotype of GS differs in biopsied African Americans versus Caucasians with hypertensive nephrosclerosis, with a marked increase in the solidified form of GS in African Americans. The association of extensive solidified GS with segmental sclerosis lesions in African Americans, but not in Caucasians, suggests different mechanisms may contribute to the development and progression of sclerosis in these two patient groups. PMID- 12081577 TI - Impaired coronary endothelial function in a rat model of spontaneous albuminuria. AB - BACKGROUND: Albuminuria is an independent risk factor of coronary artery disease and has been proposed to reflect a general endothelial disorder. The Munich Wistar Fromter (MWF) rat strain develops spontaneous albuminuria and, therefore, may be an interesting experimental model to study alterations of endothelial function under conditions of increased albuminuria. Our aim was to investigate if the MWF strain shows generalized endothelial dysfunction or endothelial dysfunction localized to the coronary vascular bed, and if so, determine which endothelial dilative mediators are involved. METHODS: Coronary and mesenteric arteries were investigated for endothelium-dependent relaxation and the contribution of prostacyclin, nitric oxide (NO) and endothelium-dependent hyperpolarization (EDH) in MWF rats compared to normal Wistar rats. In addition, as MWF rats show increased blood pressure, spontaneously hypertensive rats (SHR) with similar hypertension but without increased albuminuria also were studied. RESULTS: Maximal total endothelium-dependent relaxation of coronary arteries was strongly impaired in MWF rats (55 +/- 3%) compared to Wistar (89 +/- 5%) and SHR (89 +/- 2%) P < 0.05, respectively. The NO-mediated relaxation as well as the relaxation mediated by EDH were significantly lower in coronary arteries from MWF compared to Wistar. In mesenteric arteries of MWF the endothelium-dependent relaxation was intact. CONCLUSIONS: The strong impairment of coronary endothelium dependent relaxation in the MWF model of spontaneous albuminuria may be due to defects in production or activity of NO and EDH. The intact mesenteric endothelium-dependent relaxation suggests that increased albuminuria may not be related to generalized endothelial vasodilator dysfunction in this model. Selective impairment of coronary endothelial function in a setting of spontaneous albuminuria may be a feature of the MWF that may be employed to further study cause-effect relations between albuminuria and coronary artery disease. PMID- 12081578 TI - Renoprotective effects of losartan in diabetic nephropathy: interaction with ACE insertion/deletion genotype? AB - BACKGROUND: The beneficial short- and long-term renoprotective effects of angiotensin I-converting enzyme (ACE) inhibition are lower in albuminuric diabetic patients homozygous for the deletion compared to the insertion polymorphism of the ACE gene. In an attempt to overcome this interaction, we evaluated the short-term renoprotective effect in diabetic nephropathy of the angiotensin II receptor antagonist losartan in patients homozygous for the insertion or the deletion allele. METHODS: Fifty-four hypertensive type 1 diabetic patients with diabetic nephropathy homozygous for the insertion (I; N = 26) or the deletion (D; N = 28) allele of the ACE/ID polymorphism were included. After four weeks of washout, the patients received losartan 50 mg daily followed by 100 mg in two treatment periods each lasting two months. Patients and investigators were blinded to ACE genotypes. At baseline and in the end of the treatment periods, 24-hour blood pressure, albuminuria and glomerular filtration rate values were determined. RESULTS: At baseline, blood pressure, albuminuria and glomerular filtration rate (GFR) values were similar in the two genotype groups [II vs. DD, 1134 (238 to 5302) vs. 1451 (227 to 8129) mg/24 h, median (range); 156/82 (17/9) vs. 153/80 (17/11) mm Hg, mean (SD); and 86 (22) vs. 88 (24) mL/min/1.73 m2, respectively]. Both doses of losartan significantly lowered blood pressure, albuminuria, and GFR (P < 0.05 vs. baseline). Losartan 100 mg was more effective than 50 mg in reducing albuminuria, 51% (95% CI; 40 to 61) versus 33% (23 to 42), respectively (P < 0.01). No differences in the impact of losartan between the II and DD groups were observed: Losartan 100 mg lowered systolic/diastolic blood pressure by 12/6 and 10/4 mm Hg, whereas albuminuria decreased by 55% (35 to 68) and 46% (28 to 61), in the II and DD groups, respectively (P = NS). CONCLUSION: Our data suggest that losartan offers similar short-term renoprotective and blood pressure lowering effects in albuminuric hypertensive type 1 diabetic patients with the ACE II and DD genotypes. However, the long-term renoprotective effects remain to be evaluated. PMID- 12081579 TI - Depression as a predictor of mortality and hospitalization among hemodialysis patients in the United States and Europe. AB - BACKGROUND: Depression is not uncommon among patients with end-stage renal disease (ESRD) being treated by hemodialysis. We investigated whether risk of mortality and rate of hospitalization may be predicted from physician-diagnosed depression and patients' self-reports of depressive symptoms. METHODS: Data were analyzed from the Dialysis Outcomes and Practice Patterns Study (DOPPS) for randomly selected ESRD patients being treated by hemodialysis in the United States (142 facilities, 2855 patients) and five European countries (101 facilities, 2401 patients). The diagnosis of depression during the past year was abstracted from the medical records. In addition, the patients were asked to indicate how much of their time over the previous four weeks they had felt (1) "so down in the dumps that nothing could cheer you up" and (2) "downhearted and blue." A response of "a good bit,""most," or "all" of the time were classified as depressed. RESULTS: The prevalence of depression was nearly 20%. The relative risks of mortality and hospitalization among depressed (vs. non-depressed), adjusted for time on dialysis, age, race, socioeconomic status, comorbid indicators and country were, respectively: 1.23 and 1.11 for physician-diagnosed depression, 1.48 and 1.15 for the "so down in the dumps" question, and 1.35 and 1.11 for the "downhearted and blue" question (P < 0.05 for all six relative risks). These associations were not significantly different between US and European patients. CONCLUSIONS: Self-reported depression by two simple questions was associated with increased risks of mortality and hospitalization for hemodialysis patients. Future research needs to assess whether early identification and treatment of depression may help to improve quality of life and survival in hemodialysis patients. PMID- 12081580 TI - Prevention of obesity-linked renal disease: age-dependent effects of dietary food restriction. AB - BACKGROUND: Hyperphagic obese Zucker rats develop glomerular injury and die of renal disease, an outcome prevented by food restriction at an early age. We examined the effects of food restriction imposed at different ages on systemic, renal hemodynamic, and hormonal changes to gain insight into the mechanisms of obesity-linked glomerular injury. METHODS: At 6 weeks of age obese Zucker rats were either fed ad libitum or were restricted in food intake at various ages (6, 12, 26, or 50 weeks) to that consumed by lean Zucker rats (14 g/day). Every four weeks 24-hour urine collections, blood pressure, and venous blood samples were obtained until the end of study (60 weeks). RESULTS: Food restriction at 6 or 12 weeks of age prevented glomerular injury and hypertrophy and delayed the development of hypertension, hypercholesterolemia, and hyperinsulinemia. Food restriction at 26 weeks of age reduced proteinuria, while restriction at 50 weeks prevented further increases in proteinuria without altering pre-existing hypercholesterolemia, hypertension, or hyperinsulinemia. Hypertriglyceridemia and glomerular hyperfiltration in the obese animals were reversed at any age by food restriction. Plasma leptin levels were elevated in all obese groups. CONCLUSIONS: (1) Early food restriction provided the greatest metabolic and renal benefits; (2) glomerular injury correlated with hyperphagia-induced hyperfiltration and hypertriglyceridemia and both were prevented by food restriction; (3) hypercholesterolemia was due to an increase in LDL and/or VLDL cholesterol; and (4) leptin does not directly contribute to glomerular injury in the obese Zucker rat. PMID- 12081581 TI - Effect of dietary protein restriction on prognosis in patients with diabetic nephropathy. AB - BACKGROUND: Recent data suggest that dietary protein restriction improves survival and delays the progression to end-stage renal disease (ESRD) in non diabetic nephropathies. The purpose of our study was to determine the effect of dietary protein restriction on survival and progression to ESRD in diabetic nephropathy. METHODS: A four-year prospective, controlled trial with concealed randomization was performed comparing the effects of a low-protein diet (0.6 g/kg/day) with a usual-protein diet. The study included 82 type 1 diabetic patients with progressive diabetic nephropathy [pre-study mean decline in glomerular filtration rate (GFR) 7.1 mL/min/year (95% CI, 5.8 to 8.5)]. The main outcome measures were decline in GFR and development of ESRD or death. RESULTS: During the follow-up period the usual-protein diet group consumed 1.02 g/kg/day (95% CI; 0.95 to 1.10) as compared with 0.89 (0.83 to 0.95) in the low-protein diet group (P = 0.005). The mean declines in GFR were 3.9 mL/min/year (2.7 to 5.2) in the usual-protein diet group and 3.8 (2.8 to 4.8) in the low-protein diet group. ESRD or death occurred in 27% of patients on a usual-protein diet as compared with 10% on a low-protein diet (log-rank test; P = 0.042). The relative risk of ESRD or death was 0.23 (0.07 to 0.72) for patients assigned to a low protein diet, after an adjustment at baseline for the presence of cardiovascular disease (P = 0.01). Blood pressure and glycemic control were comparable in the two diet groups during the follow-up period. CONCLUSION: Moderate dietary protein restriction improves prognosis in type 1 diabetic patients with progressive diabetic nephropathy in addition to the beneficial effect of antihypertensive treatment. PMID- 12081582 TI - Health care utilization among patients with chronic kidney disease. AB - BACKGROUND: Higher hospitalization rates among end-stage renal disease (ESRD) patients impose a substantial burden on the U.S. health care system. Early identification of patients with chronic kidney disease (CKD) and determination of factors associated with increased morbidity may lead to appropriate interventions to attenuate the complications of CKD and possibly reduce future resource utilization. METHODS: This retrospective cohort study of CKD patients in an outpatient nephrology clinic was performed to identify risk factors for hospitalization. The study population consisted of adults with elevated serum creatinine (females > or =1.5 mg/dL, males > or =2.0 mg/dL). Hospitalizations, hospital days and outpatient nephrology visits were examined. RESULTS: Among the 259 patients, 123 (47%) were hospitalized during a median follow-up of 11.4 months. The number of hospitalizations and hospital days per patient-year at risk were 0.96 and 6.6, respectively. Cardiovascular disease/hypertension accounted for the majority of hospitalizations. In a multivariate regression analysis, older age (RR 1.01, 95% CI 1.00, 1.03) and presence of cardiac disease (RR 1.91, 95% CI 1.19, 3.07) were associated with higher risk of hospitalization while higher serum albumin (RR 0.58, 95% CI 0.35, 0.95) and higher hematocrit (RR 0.92, 95% CI 0.87, 0.97) were associated with lower risk of hospitalization. Higher serum albumin (RR 0.34, 95% CI 0.21, 0.55), higher hematocrit (RR 0.87, 95% CI 0.81, 0.93) and use of ACE-inhibitors (RR 0.63, 95% CI 0.47, 0.84) were associated with lower risk of subsequent hospital days. Erythropoietin (RR 1.47, 95% CI 1.11, 1.82) use was associated with higher risk of outpatient nephrology visits. CONCLUSION: Certain potentially modifiable factors appear to be associated with increased resource utilization. It is hypothesized that attention to these factors may lead to improved outcomes in this patient population, which could result in reduced utilization. PMID- 12081583 TI - Kidney Injury Molecule-1 (KIM-1): a novel biomarker for human renal proximal tubule injury. AB - BACKGROUND: Traditional blood and urine markers for the diagnosis of various renal diseases are insensitive and nonspecific. Kidney Injury Molecule-1 (KIM-1) is a type 1 transmembrane protein, with an immunoglobulin and mucin domain, whose expression is markedly up-regulated in the proximal tubule in the post-ischemic rat kidney. The ectodomain of KIM-1 is shed from cells. The current studies were carried out to evaluate whether KIM-1 is present in human acute renal failure and might serve as a urinary marker of acute renal tubular injury. METHODS: Kidney tissue samples from six patients with biopsy-proven acute tubular necrosis (ATN) were evaluated by immunohistochemistry for expression of KIM-1. Urine samples were collected from an additional thirty-two patients with various acute and chronic renal diseases, as well as from eight normal controls. Urinary KIM-1 protein was detected by immunoassay and was quantified by ELISA. RESULTS: There was extensive expression of KIM-1 in proximal tubule cells in biopsies from 6 of 6 patients with confirmed ATN. The normalized urinary KIM-1 levels were significantly higher in patients with ischemic ATN (2.92 +/- 0.61; N = 7) compared to levels in patients with other forms of acute renal failure (0.63 +/- 0.17, P < 0.01; N = 16) or chronic renal disease (0.72 +/- 0.37, P < 0.01; N = 9). Adjusted for age, gender, length of time delay between the initial insult and sampling of the urine, a one-unit increase in normalized KIM-1 was associated with a greater than 12-fold (OR 12.4, 95% CI 1.2 to 119) risk for the presence of ATN. Concentrations of other urinary biomarkers, including total protein, gamma glutamyltransferase, and alkaline phosphatase, did not correlate with clinical diagnostic groupings. CONCLUSIONS: A soluble form of human KIM-1 can be detected in the urine of patients with ATN and may serve as a useful biomarker for renal proximal tubule injury facilitating the early diagnosis of the disease and serving as a diagnostic discriminator. PMID- 12081584 TI - Sevelamer attenuates the progression of coronary and aortic calcification in hemodialysis patients. AB - BACKGROUND: Cardiovascular disease is frequent and severe in patients with end stage renal disease. Disorders of mineral metabolism may contribute by promoting cardiovascular calcification. METHODS: We conducted a randomized clinical trial comparing sevelamer, a non-absorbed polymer, with calcium-based phosphate binders in 200 hemodialysis patients. Study outcomes included the targeted concentrations of serum phosphorus, calcium, and intact parathyroid hormone (PTH), and calcification of the coronary arteries and thoracic aorta using a calcification score derived from electron beam tomography. RESULTS: Sevelamer and calcium provided equivalent control of serum phosphorus (end-of-study values 5.1 +/- 1.2 and 5.1 +/- 1.4 mg/dL, respectively, P = 0.33). Serum calcium concentration was significantly higher in the calcium-treated group (P = 0.002), and hypercalcemia was more common (16% vs. 5% with sevelamer, P = 0.04). More subjects in the calcium group had end-of-study intact PTH below the target of 150 to 300 pg/mL (57% vs. 30%, P = 0.001). At study completion, the median absolute calcium score in the coronary arteries and aorta increased significantly in the calcium treated subjects but not in the sevelamer-treated subjects (coronary arteries 36.6 vs. 0, P = 0.03 and aorta 75.1 vs. 0, P = 0.01, respectively). The median percent change in coronary artery (25% vs. 6%, P = 0.02) and aortic (28% vs. 5%, P = 0.02) calcium score also was significantly greater with calcium than with sevelamer. CONCLUSIONS: Compared with calcium-based phosphate binders, sevelamer is less likely to cause hypercalcemia, low levels of PTH, and progressive coronary and aortic calcification in hemodialysis patients. PMID- 12081585 TI - Role of cyclooxygenase-2 in hyperprostaglandin E syndrome/antenatal Bartter syndrome. AB - BACKGROUND: Hyperprostaglandin E syndrome/antenatal Bartter syndrome (HPS/aBS) is a congenital salt-losing tubulopathy with an induced expression of cyclooxygenase 2 (COX-2) in the macula densa probably leading to hyperreninemia. Inhibition of stimulated prostaglandin E2 (PGE2) formation with indomethacin results in a significant improvement of clinical symptoms and is therefore standard therapy. Using the COX-2 selective inhibitor rofecoxib, we investigated the role of COX-2 in the pathophysiology of HPS/aBS. METHODS: Six clinically well-characterized patients with HPS/aBS (3 girls) were enrolled into the study. Four patients had mutations in the renal potassium channel ROMK, one patient in the furosemide sensitive cotransporter NKCC2, whereas in one patient no molecular abnormality could be detected. Median age was 15.8 years (range: 9.1 to 19.0 years). Patients were evaluated on indomethacin treatment, 3 days after indomethacin withdrawal, and after 4 days of treatment with rofecoxib. Therapeutic drug monitoring was performed. RESULTS: COX-2-selectivity of rofecoxib was confirmed in vivo and ex vivo. Both indomethacin and rofecoxib ameliorated clinical symptoms, the typical laboratory findings, and significantly suppressed PGE2 and PGE-M excretion to normal values while it was elevated under withdrawal conditions. Rofecoxib suppressed hyperreninemia to a similar extent as indomethacin. CONCLUSION: In patients with HPS/aBS, excessive PGE2 synthesis and hyperreninemia is dependent on COX-2 activity. This observation proves the stimulatory role of COX-2 on renin secretion in salt-depletion in humans. Clinical long-term efficacy and potential side effects of rofecoxib need to be evaluated in a larger cohort of HPS/aBS patients. PMID- 12081586 TI - A novel interleukin-8 polymorphism is associated with severe systemic lupus erythematosus nephritis. AB - BACKGROUND: Interleukin-8 (IL-8) is a potent neutrophil chemokine that has been implicated in the pathogenesis of renal inflammation in human glomerulonephritis. To explain inter-patient variations in renal inflammation during diseases such as systemic lupus erythematosus (SLE), it was postulated that the promoter region of the IL-8 gene contains polymorphic residues that influence the level of IL-8 expression in response to immune-complex deposition, and thereby affect the severity of renal injury. This study was undertaken to identify polymorphisms in the 5'-flanking region of the IL-8 gene that correlate with the severity of SLE nephritis. METHODS: A 1526 base pair segment of the IL-8 5'-flanking region was PCR amplified from the genomic DNA of 100 individuals and sequenced on an automated capillary electrophoresis system. Sequence data were compared with the published IL-8 sequence to identify polymorphisms. Allelic variations were verified by cloning and re-sequencing, and also by restriction enzyme analysis. Patients with SLE nephritis were genotyped for IL-8 polymorphisms, and associations between specific alleles and severity of SLE nephritis [based on the World Health Organization (WHO) classification] were determined. RESULTS: Three single nucleotide polymorphisms were identified in the IL-8 flanking region. Labeled relative to the IL-8 translational start site, these are T-845C, T-738A, and A-353T. T-845C and T-738A are novel, and found primarily in African Americans. The C for T change at position -845 was found to be 3.6 to 7.5 times more frequent in African Americans with severe (WHO Class IV) SLE nephritis, than in African American controls, or patients with less severe forms of SLE nephritis, respectively. CONCLUSIONS: IL-8-845C might predispose African Americans with SLE nephritis to more severe renal injury, perhaps by influencing IL-8 expression. Genotyping patients with glomerulonephritis for IL-8 polymorphisms may be useful in predicting disease outcome and individualizing immunosuppressive therapy. PMID- 12081587 TI - Effects of feeding on albumin synthesis in hypoalbuminemic hemodialysis patients. AB - BACKGROUND: Hypoalbuminemia is a powerful predictor of morbidity and mortality in hemodialysis (HD) patients and results from a reduction in albumin synthesis. It is not known if this is associated with any impairment of the normal response to feeding. METHODS: Protein turnover and albumin synthesis were measured in the fasting and fed state using a primed constant infusion of L-[1-(13)C]leucine in seven hypoalbuminemic (albumin < or = 36 g/L) HD patients (HHD), seven normoalbuminemic (albumin > or = 40 g/L) HD patients (NHD) and nine age-matched normal controls. RESULTS: The increase in albumin synthesis on feeding was impaired in HHD patients (fasting 15.0 +/- 1.5 vs. fed 17.7 +/- 2.9%, P = NS) compared to NHD (fasting 13.7 +/- 0.9 vs. fed 17.4 +/- 1.0%, P < 0.05) and controls (fasting 12.9 +/- 0.6 vs. fed 15.2 +/- 0.6%, P < 0.05). In addition, body mass index and percent body fat were significantly (P < 0.05) lower in HHD (20.8 +/- 1.3 kg/m2, 23.4 +/- 2.0%) than NHD (26.7 +/- 1.3 kg/m2, 33.1 +/- 3.2%) or controls (26.2 +/- 1.1 kg/m2, 32.6 +/- 1.8%). There was no difference in dietary protein or energy intake in the three groups. CONCLUSIONS: There are differences of body composition and protein metabolism in HHD patients that may be related to an impaired metabolic response to feeding. PMID- 12081588 TI - Proactive monitoring of pediatric hemodialysis vascular access: effects of ultrasound dilution on thrombosis rates. AB - BACKGROUND: Hemodialysis vascular access thrombosis (VAT) is a significant cause of morbidity for hemodialysis patients and results, in part, from decreased access flow potentially caused by venous outflow stenosis. We have previously shown ultrasound dilution (UD) to be a practical and reliable predictor of venous outflow in children receiving hemodialysis. METHODS: The current study is the first to our knowledge to assess the impact of a proactive UD monitoring program upon VAT in pediatric patients. Nine patients experienced 18 VAT over the two year study. Mean values for variables potentially associated with VAT were compared to values from a size-matched seven patient group without VAT during the study period. VAT rates were compared between the year-before (pre-UD era) and year-after (UD era) UD was initiated. During the latter half of the UD era (rapid referral period), patients with VA flow rate (QAcorr) <650 mL/min/1.73 m2 were referred for balloon angioplasty within 48 hours. RESULTS: Mean QAcorr was lower for patients with subsequent VAT (562 +/- 290 mL/min/1.73 m2) versus patients without VAT (1005 +/- 372 mL/min/1.73 m2; P = 0.02). The VAT rate was significantly lower in the UD era (4.1 VAT/100 patient-months) versus the pre-UD era (11.0 VAT/100 patient-months; P = 0.03). The decrease in VAT rates was caused predominantly in the rapid referral period, where the VAT rate dropped to 0.96 VAT/100 patient-months (P < 0.001). Cost of vascular access management was 65% higher ($1264 vs. $765/patient-month) in the pre-UD era, reflecting the increased cost for treatment of VAT. CONCLUSIONS: Monthly QAcorr <650 mL/min/1.73 m2 is predictive of imminent VAT in children receiving hemodialysis. Prompt referral for angioplasty of VA with QAcorr <650 mL/min/1.73 m2 leads to decreased VAT rates in children. PMID- 12081589 TI - Serum prealbumin is higher in peritoneal dialysis than in hemodialysis: a meta analysis. AB - BACKGROUND: Although not widely appreciated, the reported concentration of serum prealbumin, like that of serum cholesterol, tends to be higher in patients on peritoneal dialysis (PD) than on hemodialysis (HD), despite the substantial loss of protein during PD. METHODS: The mean difference in serum prealbumin was quantified by meta-analysis of the mean differences found in six cohorts with both PD and HD patients (set 1; N = 639) using a fixed-effects model, and meta analysis of the mean prealbumin values reported in 23 cohorts of unselected dialysis patients on a single modality (set 2; 9 PD cohorts, 14 HD cohorts; N = 12,256) using a mixed model. For comparison, the mean difference in serum albumin concentration between PD and HD also was estimated in sets 1 and 2 using the same methods. RESULTS: In set 1, the mean prealbumin difference (PD-HD) in the individual cohorts ranged from 3.6 to 14.7 mg/dL (P < 0.05 in five cohorts), and the weighted mean difference was 5.4 mg/dL (95% CI, 3.8 to 7.0 mg/dL). In set 2, weighted mean prealbumin was 8.1 mg/dL (95% CI, 5.2 to 10.9 mg/dL) higher in PD than in HD in the entire data set, and 6.9 mg/dL (95% CI, 5.2 to 8.6 mg/dL) higher in a sensitivity analysis that excluded two outlying HD studies. By contrast, weighted mean serum albumin concentration was significantly lower in PD than in HD in both sets 1 and 2; the mean difference was 0.25 g/dL (95% CI, 0.14 to 0.36 g/dL) in set 1 and 0.28 g/dL (95% CI, 0.14 to 0.42 g/dL) in set 2. CONCLUSIONS: Serum prealbumin level is approximately 6 mg/dL higher in PD than HD, perhaps due to the stimulation of hepatic synthesis by PD albumin loss, while serum albumin is approximately 0.3 g/dL lower in PD. Different reference ranges and clinical targets (such as, K/DOQI guidelines) are needed for PD and for HD. PMID- 12081590 TI - Randomized comparison of split tip versus step tip high-flow hemodialysis catheters. AB - BACKGROUND: Our purpose was to compare the function and complications of two high flow polyurethane hemodialysis catheters. METHODS: This prospective, randomized trial compared the Ash-Split (MedComp) and Opti-Flow (Bard Access Systems) catheters. All patients referred for tunneled hemodialysis catheter placement were offered entry in the study, provided they met inclusion criteria. Catheters were placed by interventional radiologists using ultrasound and fluoroscopic guidance. Procedure time and initial complications were recorded. Effective (QbEff) catheter flow rates and recirculation were studied at baseline, one month, three and six months using ultrasonic dilution (Transonic) at various pump speeds (Qb). Episodes of catheter malfunction and infection were recorded. Catheter removal or six months was the study endpoint. RESULTS: A total of 132 patients were enrolled in the trial. The groups did not differ as to age, sex distribution, height or weight (P> 0.05). Initial complications included kinking resulting in catheter failure (Optiflow N = 3), and tunnel bleeding (Optiflow N = 1; Ash N = 3). Adjusted mean flow rates (QbEff) at Qb300 were 299 mL/min Ash and 305 mL/min Optiflow (P = 0.06), at Qb400 were 365 mL/min Ash and 382 mL/min Optiflow (P = 0.01), and at QbMax were 414 mL/min Ash and 433 mL/min Optiflow (P = 0.03). Recirculation was significantly higher with the Optiflow catheter at most measurement points. Total late complications were lower in the Ash group (P = 0.04), and catheter survival was significantly higher in the Ash group (P = 0.02). CONCLUSIONS: Both catheters can deliver flow rates well beyond those recommended by the Dialysis Outcomes Quality Initiative. While the Optiflow delivered higher flow rates at some measurement points, this was offset by higher recirculation. The Ash catheter showed a long-term survival advantage and fewer late complications. PMID- 12081591 TI - Absence of donor MHC antigen expression ameliorates chronic kidney allograft rejection. AB - BACKGROUND: In previous studies, we have demonstrated that a subset of mouse kidney allografts has prolonged survival without any immunosuppressive treatment. Chronic rejection (CR) develops in these long surviving grafts. The pathologic features of CR in this model are similar to CR in human kidney grafts. METHODS: To explore the role of donor major histocompatibility complex (MHC) antigens in the development of CR, we performed vascularized kidney transplants using kidneys from donor mice that lack expression of both MHC class I and II antigens (MHC-/ ). RESULTS: Survival was significantly improved in recipients of MHC-/- allografts. This enhanced survival was associated with higher glomerular filtration rate (GFR) in MHC-/- allografts (4.92 +/- 0.54 cc/min/kg) compared to controls (2.19 +/- 0.63 cc/min/kg; P = 0.004). The typical histologic features of CR were markedly reduced in MHC-/- allografts. Semiquantitative histopathological scores for MHC-/- grafts (13.3 +/- 2.1) were significantly lower than in control allografts (19.0 +/- 1.0; P = 0.04). Along with this improvement in structural abnormalities, significantly fewer CD4+ T (38.3 cells/mm(2) vs. 75.0 cells/mm(2); P = 0.008), CD8+ T cells (38.7 vs. 96 cells/mm(2), respectively; P = 0.008) and macrophages (60 vs. 134 cells/mm(2), respectively; P = 0.04) infiltrated MHC-/- allografts compared to controls. The levels of intragraft cytokine mRNA expression also were reduced in MHC-/- allografts compared to control allografts. Finally, serum alloantibodies were virtually undetectable in recipients of MHC-/- kidney allografts. CONCLUSIONS: Cell surface expression of donor MHC antigens promotes the development of CR. Donor antigen expression promotes the accumulation of infiltrating cells in the graft and the development of donor specific alloantibodies. Abrogation of these responses is associated with improved graft survival and reduced CR in MHC-/- grafts. PMID- 12081592 TI - Advanced glycation end products and mortality in hemodialysis patients. AB - BACKGROUND: Advanced glycation and oxidation end products (AGEs) cause oxidative stress and trigger cytokine driven inflammatory reactions in vitro. The net effects on markers of inflammation and acute phase proteins in vivo as well as their influence on survival in hemodialysis patients are unknown. METHODS: We conducted a cross-sectional study in 312 stable hemodialysis patients and analyzed the interrelationships of AGEs and C-reactive protein (CRP) and their predictive effect on all-cause as well as cardiovascular mortality. Mortality was monitored prospectively over a period of 32 months. AGEs were determined by measuring total serum fluorescent AGEs (AGE-fl) and Nepsilon-(carboxymethyl) lysine (CML). RESULTS: The levels of AGE-fl, CML and CRP were 3.2-, 3.8- and 10 fold higher as compared to healthy controls. AGE-fl and CML levels correlated significantly with each other but not with CRP or serum albumin. Patients with high (above median values) AGE-fl or CML levels (109 x 103 AU and 1.4 microg/mL, respectively) had a significant better survival than those with low (below median values) AGE-fl or CML levels. Patients with high CRP levels (above 7.7 mg/L = median value) had a better survival than those with low CRP (below median value) when AGE-fl or CML levels were high in parallel. CONCLUSIONS: In contrast to in vitro data and to current hypotheses, the presence of high serum AGEs, as measured by AGE-fl and CML, were not linked to increased mortality. Statistically, high serum AGEs partly overcame the negative impact of the acute phase response on mortality in hemodialysis patients. Whether the benefit of high serum AGEs is an epiphenomenon or reflects a better nutritional support needs further studies. PMID- 12081593 TI - Post-transplant renal function in the first year predicts long-term kidney transplant survival. AB - BACKGROUND: Improvements in long-term kidney graft survival have been recently noted. However, the reasons for this were unclear. This study examined post transplant renal function within the first year as an independent variable influencing long-term survival. METHODS: The influence of demographic characteristics (age, sex, race); transplant variables (cadaver versus living donor, cold ischemia time, HLA mismatching, delayed graft function and transplant year), and post-transplant variables (immunosuppressive agents for the prevention of acute rejection, clinical acute rejection and post-transplant renal function in the first year) on graft survival were analyzed for 105,742 adult renal transplants between 1988 and 1998. Renal function in the first year was expressed as serum creatinine at six months and one year and delta creatinine (change in serum creatinine between 6 months and 1 year). Graft half-life was used to measure long-term survival. RESULTS: During this 11-year period, the one-year serum creatinine values for cadaver recipients steadily improved, from 1.82 +/- 0.82 mg/dL in 1988 to 1.67 +/- 0.82 mg/dL in 1998 (P < 0.001), as did the graft half-life. There was a progressive decline in graft half-life for each incremental increase of six month, one year and Delta creatinine for living and cadaver donor transplants as well for cadaver transplants with donor age > and < or =50 years. The Relative Hazard (RH) for graft failure was 1.63 (1.61, 1.65; P < 0.0001) with each increment of 1.0 mg/dL of serum creatinine at one year post transplant and it increased to 2.26 (2.2, 2.31; P < 0.0001) when the Delta creatinine was 0.5 mg/dL. The RH reduction for graft failure was substantially lower for the years 1993, 1996, 1997 and 1998 when post-transplant renal function was not included in the model (P < 0.05). However, the RH reduction per year was not different when post-transplant creatinine was included in the model, 1.01 (0.94 to 1.05; P = 0.89). CONCLUSION: In conclusion, one-year creatinine and Delta creatinine values predict long-term renal graft survival. Recent improvements in graft half-life are related to conservation of renal function within the first year post-transplantation. PMID- 12081594 TI - Immune responsiveness in renal transplant recipients: mycophenolic acid severely depresses humoral immunity in vivo. AB - BACKGROUND: Current immunosuppressive drug treatments for renal transplant recipients result in high one-year graft survival rates. Despite adequate suppression of the immune response directed to the allograft, the immune system remains able to cope with many infectious agents. METHODS: To define the influence of distinct immunosuppressive treatment protocols, primary and secondary cellular and humoral immune responses in groups of renal transplant recipients were studied: patient treated with prednisolone and cyclosporine A (P/CsA); with IgA CD3 monoclonal antibody as a rejection treatment superimposed on prednisolone and cyclosporine A (IgA CD3 mAb+P/CsA); and with prednisolone, cyclosporine A and mycophenolate mofetil (P/CsA/MMF). RESULTS: Primary in vitro proliferative responses to the protein antigen keyhole limpet hemocyanin (KLH) were not significantly disturbed in P/CsA treated patients, or in IgA CD3 mAb+P/CsA and P/CsA/MMF treated patients. In vitro proliferative responses to the recall antigen tetanus toxoid (TT) were similarly unaffected. Antigen-specific antibody responses to immunization with KLH and TT were not affected by treatment with P/CsA, or by IgA CD3 mAb+P/CsA, but were severely disturbed in patients treated with P/CsA/MMF. All patients displayed a profound inhibition of the delayed-type hypersensitivity skin reactivity to KLH and recall antigens. Nevertheless, in most patients with P/CsA treatment, T cell infiltrates were observed in skin biopsies from the site of KLH challenge, while expression of intercellular cell adhesion molecule-1 (ICAM-1) expression in challenged skin was significantly decreased in these patients. The balance between T helper 1 and T helper 2 cells was unaffected by immunosuppressive treatments during one year of follow-up. CONCLUSIONS: Immunosuppressive drug treatment with P/CsA inhibits delayed-type hypersensitivity skin reactions to both primary and frequently encountered antigens. Histological studies indicate an effect on ICAM-1 expression, leaving the influx of CD3pos T cells unaffected. Administration of a 10-day course of IgA CD3 mAb does not add profound immunosuppressive effects on the measured parameters. In contrast, addition of treatment with MMF profoundly decreases both primary and secondary humoral immune responsiveness in vivo. Finally, no effect of the studied immunosuppressive drugs on Th1/Th2 balance in vivo was measured. PMID- 12081595 TI - Tailoring the initial vascular access for dialysis patients. AB - BACKGROUND: Creating a functioning initial arteriovenous (AV) access for aging and diabetic end-stage renal disease (ESRD) hemodialysis patients has been a challenge. METHODS: This study describes 748 consecutive primary AV access creations and their primary (unassisted) and secondary (assisted) access survival at a single center. Twenty-four percent of the patients had diabetes as their cause of ESRD and the average age was 59.6 years. No patient receiving an initial AV access required synthetic graft material. All received an AV fistula. Three types of fistulae were created and their distribution varied significantly for diabetic and non-diabetic patients (respective percentages): forearm AV fistula (24%, 62%), perforating vein fistula (PVF) at the elbow (48%, 21%) and non-PVF at the elbow (29%, 17%). RESULTS: Results of access survival for age groups <65 and 65+ years, male and female, diabetic and non-diabetic subgroups ranged from 51 to 75% for unassisted and from 75 to 96% for assisted two year access survival. PVF appeared to be advantageous over non-PVF access at the elbow. First intervention for peripheral steal syndrome was required at a rate of 7 and 0.6 per 100 patient years at risk for diabetic and non-diabetic patients, respectively. The thrombosis rates per patient year of 0.03 for non-diabetics and 0.07 for diabetics are superior to previously published results for AV fistulae or for a combined AV fistula-AV graft approach. CONCLUSIONS: Potential explanations for these excellent results among elderly and diabetic patients include preoperative evaluation, exclusive use of native vessels, a variable surgical approach including PVF, and the experience of a single operator. PMID- 12081596 TI - Left ventricular hypertrophy, cardiac remodeling and asymmetric dimethylarginine (ADMA) in hemodialysis patients. AB - BACKGROUND: The endogenous inhibitor of nitric oxide (NO), asymmetric dimethylarginine (ADMA), is a strong predictor of adverse cardiovascular outcomes in patients with end-stage renal disease (ESRD). METHODS: Since arterial and cardiac remodeling is associated with altered endothelial microcirculatory responses to forearm ischemia (a NO-dependent response), interference of ADMA with the NO system may be important for the pathogenesis of left ventricular hypertrophy (LVH) in these patients. This study sought to identify the relationship between plasma ADMA and LV geometry and function in a cohort of 198 hemodialysis patients. RESULTS: Plasma ADMA was significantly higher (P = 0.008) in patients with LVH (median 3.00 micromol/L, inter-quartile range 1.73 to 3.97 micromol/L) than in those without this alteration (1.88 micromol/L, 1.15 to 3.56 micromol/L) and was significantly related to left ventricular (LV) mass (r = 0.26, P < 0.001). Interestingly, ADMA was much higher (P < 0.001) in patients with concentric LVH (3.60 micromol/L, 2.90 to 4.33 micromol/L) than in patients with eccentric LVH (2.17 micromol/L, 1.47 to 3.24 micromol/L) or normal LV mass (1.76 micromol/L, 1.13 to 2.65 micromol/L). Furthermore, plasma ADMA was higher (P = 0.02) in patients with systolic dysfunction (3.52 micromol/L, 2.08 to 5.87 micromol/L) than in those with normal LV function (2.58 micromol/L, 1.53 to 3.84 micromol/L) and inversely related to ejection fraction (EF; r = -0.25, P < 0.001). The link between ADMA and LV mass and EF was confirmed by multivariate analysis (ADMA vs. LVMI, beta = 0.17, P = 0.006; ADMA vs. EF, beta = -0.24, P < 0.001). CONCLUSIONS: Overall, this study indicates that raised plasma concentration of ADMA is associated to concentric LVH and LV dysfunction. Intervention studies are needed to see whether the link between ADMA and concentric LVH remodeling and LV dysfunction is a causal one. PMID- 12081597 TI - Interleukin-1beta as a mediator of tubulointerstitial fibrosis. PMID- 12081598 TI - Osteopontin in cyclosporine toxicity. PMID- 12081599 TI - Fragmentation of filtered proteins and implications for glomerular protein sieving in Fanconi syndrome. PMID- 12081600 TI - End-stage renal disease in developing countries. PMID- 12081601 TI - Occult hepatitis B virus infection and its clinical implications. AB - Occult hepatitis B virus (HBV) infection is characterized by presence of HBV infection with undetectable hepatitis B surface antigen (HBsAg). Serum HBV level is usually less than 104 copies/mL in these patients. Diagnosis of occult HBV infection requires sensitive HBV-DNA PCR assay. Several possibilities have been hypothesized as the mechanisms of occult HBV infection. These include: (i) mutations of HBV-DNA sequence; (ii) integration of HBV-DNA into host's chromosomes; (iii) infection of peripheral blood mononuclear cells by HBV; (iv) formation of HBV-containing immune complex; (v) altered host immune response; and (vi) interference of HBV by other viruses. The precise prevalence of occult HBV infection remains to be defined. The clinical implications of occult HBV infection involve different clinical aspects. First of all, occult HBV infection harbours potential risk of HBV transmission through blood transfusion, haemodialysis, and organ transplantation. Second, it may serve as the cause of cryptogenic liver disease, contribute to acute exacerbation of chronic hepatitis B, or even fulminant hepatitis. Third, it is associated with development of hepatocellular carcinoma. Fourth, it may affect disease progression and treatment response of chronic hepatitis C. Most of the previous studies utilized retrospective observation without control groups, and lacked direct association of occult HBV infection with specific pathological changes and disease progression. Highly sensitive, quantitative, and functional molecular analyses of HBV, combined with a well-designed prospective clinical assessment will provide the best approach for the future study of occult HBV infection. PMID- 12081602 TI - T-cell antigenic determinants within hepatitis C virus nonstructural protein 3 and cytokine production profiles in hepatitis C. AB - The aim of this study was to further investigate the role of T-helper cells in hepatitis C virus (HCV) infection, focusing on the T-cell antigenic determinants and cytokine profiles of nonstructural 3 (NS3) protein-stimulated peripheral blood mononuclear cells (PBMCs) of HCV patients. A total of 12 recombinant proteins of theNS3 region were purified and used to test T-cell proliferative response and antigenic determinants of HCV-seropositive patients. In addition, cytokines produced by antigen stimulated PBMCs were measured. Our data showed that PBMCs from 55.7% (34/61) of HCV patients proliferated to at least one antigen, but PBMCs of HCV seronegative patients did not. In addition, PBMCs from about 82.0% (32/39) HCV-seropositive patients produced significant amounts of cytokines (10 pg/mL). Interestingly, PBMCs from 66% of patients produced TH2 related cytokines such as interleukin (IL)-4 and IL-5. In mappingexperiments, the data showed multiple T-cell antigenic determinants. Our data demonstrated that NS3 antigen-stimulated PBMCs of HCV patients recognized multiple T-cell antigenic determinants and produced significant amounts of TH0 or TH2-related cytokines, which might play a critical role in the chronicity of HCV infection. PMID- 12081603 TI - In vitro infection of human peripheral blood mononuclear cells by a defective hepatitis B virus with a deletion in the PreS1 region of the viral genome. AB - Previously, we identified a defective hepatitis B virus (HBV) which contains a 183 nucleotide deletion in the PreS1 region of the viral genome affecting the S gene promoter in sera from hepatitis B surface antigen (HBsAg)-negative patients with serum HBV-DNA. The aim of this study was to analyse the infectivity of this mutant. Peripheral blood mononuclear cells (PBMC) from a healthy donor were incubated with serum samples from 2 HBsAg-negative patients with serum HBV-DNA (infected with wild-type and deletion mutant HBV), from an HBsAg carrier (infected with wild-type HBV) and from a healthy donor. After 1 week, HBV-DNA was detected by polymerase chain reaction (PCR) in all supernatants and cells incubated with the HBV-DNA-positive inocula. DNase and trypsin pretreatment confirmed intracellular localization of HBV-DNA in cells. HBV-RNA and covalently closed circular HBV-DNA were also detected in PBMC, indicating that the viral DNA infecting these cells was transcriptionally active. Deletion mutant and wild-type HBV were detected in the supernatants and cells infected with the two HBsAg negative sera, while only wild-type HBV was detected in the supernatant and cells incubated with the serum from the HBsAg-carrier. In conclusion, this HBV deletion mutant can infect, replicate and release viral particles in in vitro infected PBMC. PMID- 12081604 TI - Replication of hepatitis B in HBsAg-positive siblings. AB - Persistent hepatitis B virus (HBV) replication is important for progression of chronic liver diseases. To understand whether there is a trend of HBV replication in siblings or not, 1850 relatives of patients with hepatocellular carcinoma (HCC) were examined prospectively for liver function test, viral markers and HBV DNA. The prevalence of HBsAg in the parents', siblings', children's and grandchildren's generations were 43.4%, 57.2%, 35.5% and 32.1%, respectively. The prevalence of hepatitis B e antigen (HBeAg) in sibling's generation (mean age 44.4 years) was 19%, which is higher than that of asymptomatic HBsAg carriers. For siblings in the children's generation, the prevalence of HBeAg in hepatitis B surface antigen (HBsAg) carriers declined from 40% in the eldest siblings to 19% in the youngest siblings. In 75 families clustered with three or more HBsAg carrier siblings, the mean age for seven families of which all siblings remained HBeAg + was younger, whereas the mean age for 35 families of which all siblings had cleared HBeAg was older. For the remaining 33 families, in only 10 families had the older siblings cleared the HBeAg earlier than the younger siblings. Twenty families showed that younger siblings cleared the HBeAg earlier than the older or middle siblings. We concluded that HBV replication in HCC relatives cannot be explained by familial tendency alone. A significant number of younger siblings appeared to have a shorter HBV replication phase than their older siblings. The possible role of this in maternal-fetal transmission is discussed. PMID- 12081605 TI - Thymosin-alpha1 and famciclovir combination therapy activates T-cell response in patients with chronic hepatitis B virus infection in immune-tolerant phase. AB - We examined whether combination therapy with thymosin-alpha1 and famciclovir would induce hepatitis B e antigen seroconversion in patients with chronic hepatitis B infection in the immune-tolerant phase without inducing significant hepatic necro-inflammation. We studied 32 hepatitis B e antigen positive patients in the immune-tolerant phase of infection, treated with 26-weeks combination therapy of famciclovir and thymosin-alpha1 (group 1). Thirty-two patients who received 26-weeks famciclovir monotherapy (group 2) and another 32 patients who received no treatment (group 3), served as controls. Peripheral blood mononuclear cell proliferation and cytokine secretion in response to recombinant HBV core and surface antigen and serial serum HBV-DNA, were assayed. No significant difference in adverse events were observed among the three groups. By week 26, the median reduction in group 1 (0.94 log10 copies/mL) was greater than group 2 (0.70 log10 copies/mL, P < 0.001). Five (15.6%) patients in group 1 at 52 weeks (median range 13-78 weeks) and none in group 2 or 3 experienced hepatitis B e antigen seroconversion (P = 0.053). Sustained serological clearance of hepatitis B e antigen was associated with activation of CD4 positive HBV-specific T-cell reactivity and were of T-helper 1. Hence combination therapy with immunomodulatory agents and nucleoside analogues should be explored. PMID- 12081606 TI - Alcohol abstinence does not offset the strong negative effect of lifetime alcohol consumption on the outcome of interferon therapy. AB - Heavy alcohol consumption has been reported to negatively affect the outcome of interferon therapy. We studied the impact of lifetime alcohol consumption in patients with chronic hepatitis C treated with interferon after 6 months of alcohol withdrawal. Alcohol intake was measured when patients with chronic hepatitis C were referred to us for the first time, and from that moment complete abstinence was recommended. After 6 months of abstinence, 150 patients with persistent elevated serum alanine aminotransferase (ALT) have been treated with interferon (IFN)-alpha, 3 or 6 microU three times per week for 12 months. Univariate and multivariate analysis were performed to identify the predictors of treatment response. Carbohydrate-deficient transferrin was employed to assess alcoholic abstinence. The sustained response rate felt from 33% in nondrinkers to 20% of mild-drinkers and to only 9% in heavy drinkers. Drinker patients showed a relapse rate twice as high as that of nondrinkers. According to the multivariate analysis, the strongest independent predictors of nonresponse were genotype 1b infection, age of the patients and their lifetime alcohol intake. Carbohydrate deficient transferrin detected at baseline, at 3 months of therapy and at the end of follow-up gave a positive result only in eight determinations (1.77%), confirming the compliance of patients to our recommendation of alcohol abstinence. Lifetime alcohol consumption has a strong negative effect on the outcome of interferon treatment, mainly in heavy drinkers. A 6-month period of abstinence may not be sufficient to offset this negative effect on treatment outcome. PMID- 12081607 TI - Fatigue in patients with chronic hepatitis C. AB - In numerous studies of symptoms in patients with chronic hepatitis C there has been no systematic assessment of both fatigue and extrahepatic manifestations. Our objective was to assess the prevalence of fatigue in patients with hepatitis C virus (HCV) infection, and to identify associations between fatigue and clinical and biological hepatic and extrahepatic manifestations. We studied 1614 patients. Data were prospectively recorded during the first visit of patients infected with HCV and the prevalence of fatigue and its association with dermatological, rheumatological, neurological and nephrological manifestations; diabetes; arterial hypertension; auto-antibodies, and cryoglobulinaemia were assessed. Then, using multivariate analysis, we identified demographic, biochemical, immunological, virological, and histological factors associated with the presence of fatigue. Fatigue was present in 53% of patients (95% confidence interval 51-56). In 17% of patients (95% confidence interval 15-19) fatigue was severe, impairing activity. Five other extrahepatic manifestations had a prevalence above 10% including, in decreasing order: arthralgia, paresthesia, myalgia, pruritus, and sicca syndrome. In univariate and multivariate analyses, fatigue, in comparison with the absence of fatigue, was associated with female gender, age over 50 years, cirrhosis, depression and purpura. Independent of these associations, fatigue was associated with arthralgia, myalgia, paresthesia, sicca syndrome and pruritus. The prevalence of fibromyalgia (as defined by the association of fatigue with arthralgia or myalgia) was 19% (95% confidence interval 17-21). There was no significant association between fatigue and the following characteristics: viral load or genotype, alcohol consumption, abnormal thyroid function, and type and level of cryoglobulinaemia. Hence, fatigue is the most frequent extrahepatic manifestation in patients infected with HCV. Fatigue is independently associated with female gender, age over 50 years, cirrhosis, depression and purpura. PMID- 12081608 TI - Antiviral treatment responses in patients with chronic hepatitis C virus infection evaluated by a third generation anti-hepatitis C virus assay. AB - Antibodies to hepatitis C virus (HCV) may decrease or disappear after viral clearance in treated or spontaneously resolved infection. We evaluated the usefulness of serial antibody assays in predicting the antiviral treatment responses. One hundred and four chronic hepatitis C patients who received 24 weeks of interferon and ribavirin combination therapy were assayed with a third generation enzyme immunoassay anti-HCV. The mean titre of anti-HCV decreased by more than 50% (from 89.5 +/- 10.8 to 43.6 +/- 17.5) at 48 weeks post-treatment in patients with a sustained virological response, while in nonsustained virological responders and nonresponders, the titres remained over 85% of the pretreatment level at 48 weeks post-treatment. There was a divergence of anti-HCV titres between sustained and nonsustained virological responders during 6-9 months. By using the ratio of 9-month to 6-month titres as an index and receiver operator characteristic curve analysis with the cut-off point set at 90%, we could differentiate sustained virological responders from nonsustained virological responders with a sensitivity and specificity of 91.7% and 90.9%, respectively, 3 months after treatment. The titre of this third generation anti-HCV decreased progressively in sustained virological responders and this assay may be used to monitor and predict antiviral treatment responses. PMID- 12081609 TI - Prevalence and risk factors for intrafamilial transmission of hepatitis C virus in Karachi, Pakistan. AB - Household contacts of hepatitis C virus (HCV)-positive patients are considered at increased risk of HCV infection. This cross-sectional study during April through June 1999 assessed the prevalence and risk behaviours associated with HCV seropositivity among the household contacts of HCV seropositive thalassaemic children in Karachi, Pakistan. Among the 341 household contacts of 86 thalassaemic HCV seropositive children who were tested, 70 (20.5%) were positive for anti-HCV antibodies. The stratified analysis showed that HCV seroprevalence among the contacts did not differ significantly by the gender of the index patient and the type of relationship of contact with the index patient. However, HCV seroprevalences among the fathers and mothers of male index patients was substantially higher compared to those of female index patients. HCV RNA was recovered and genotyped from nine index patients and corresponding nine HCV seropositive household contacts. HCV genotype 3a and 3b were found in 89% (8/9) and 11% (1/9) of the pairs, respectively. The final multivariable conditional logistic regression model revealed that after adjusting for the effect of ethnicity and past hospital admission history, the HCV-seropositive household contacts were more likely than HCV seronegative household contacts to have been bitten by the carrier [adjusted matched odds ratio (mOR)=2.6, 95% CI 1.3-5.2] or have shared a toothbrush with the carrier (adjusted mOR=8.2; 95% CI 1.56-43.5). Control efforts should focus on the risk behaviours. PMID- 12081610 TI - Changing scenario of hepatitis A virus and hepatitis E virus exposure among the primitive tribes of Andaman and Nicobar Islands, India over the 10-year period 1989-99. AB - The Andaman and Nicobar Islands, Union Territory of India, are home to six primitive tribes. Studies carried out earlier among these tribes revealed very high rates of hepatitis B infection. We have now studied hepatitis A and E infection among them. A total of 951 serum samples were collected from four accessible tribes (Nicobarese, Shompens, Onges and Great Andamanese) and tested for antibodies against hepatitis A and E viruses. In addition, 240 serum samples collected a decade earlier from age-stratified Nicobarese were also screened. Hepatitis A virus (HAV) infection was found to be highly endemic among all the tribes, whereas hepatitis E virus (HEV) infection was common among the Nicobarese and Shompens. The age group-wise prevalence of these infections among the Nicobarese showed different patterns, HAV prevalence rising significantly from those aged 10 years and thereafter reaching a plateau, whereas HEV prevalence was found to be more evenly distributed over all age groups, but rising somewhat after 30 years of age. Over the last decade, the prevalence of HAV among the Nicobarese has declined slightly, particularly in those aged 10 years or less whereas HEV infection has more than doubled over all age ranges. Different HEV prevalence observed among the tribes could not be attributed to differences in sanitation or water supply. This fact and the different age-wise patterns of HAV and HEV prevalences is suggestive of different modes of transmission of HEV that are not shared. The highest rates for HEV were among those tribes which reared pigs suggesting that pigs might serve as reservoir of HEV. Further studies are needed, however, to validate these findings. PMID- 12081611 TI - Pharmacokinetics and pharmacokinetic/pharmacodynamic integration of marbofloxacin in calf serum, exudate and transudate. AB - Marbofloxacin is a fluoroquinolone antimicrobial drug used in cattle for the treatment of respiratory infections. In this investigation the pharmacokinetics (PK) of marbofloxacin were determined after intravenous and intramuscular dosing at a dosage of 2 mg/kg. In addition the ex vivo pharmacodynamics (PD) of the drug were determined in serum and three types of tissue cage fluid (transudate, inflammatory exudate generated by carrageenan and exudate generated by lipopolysaccharide). Marbofloxacin PK was characterized by a high volume of distribution after dosing by both routes (1.28 L/kg intravenous and 1.25 L/kg intramuscular). Corresponding area under the concentration-time curve (AUC) and elimination half-life (t(1/2)el) values were 9.99 and 10.11 microg h/mL and 4.23 and 4.33 h, respectively. Values of AUC for carrageenan-induced exudate, lipopolysaccharide-induced exudate and transudate were, respectively, 8.28, 7.83 and 7.75 microg h/mL after intravenous and 8.84, 8.53 and 8.52 microg h/mL after intramuscular dosing. Maximum concentration (Cmax) values were similar for the three tissue cage fluids after intravenous and intramuscular dosing. For in vivo PK data values of AUC: minimum inhibitory concentration (MIC) (AUIC) ratio for serum were 250 and 253, respectively, after intravenous and intramuscular dosing of marbofloxacin against a pathogenic strain of Mannheimia haemolytica (MIC=0.04 microg/mL). For all tissue cage fluids AUIC values were >194 and >213 after intravenous and intramuscular dosing, and Cmax/MIC ratios were 9 or greater, indicating a likely high level of effectiveness in clinical infections caused by M. haemolytica of MIC 0.04 microg/mL or less. This was confirmed by both in vitro (serum) and ex vivo (serum, exudate and transudate) measurements, which demonstrated a concentration-dependent killing profile for marbofloxacin against M. haemolytica. Ex vivo, after 24-h incubation, virtually all bacteria were killed (<10 cfu/mL) in all samples collected up to 9 h (serum), 24 h (carrageenan induced exudate and transudate) and 36 h (lipopolysaccharide-induced exudate). Application of the sigmoid Emax equation to the ex vivo antibacterial data provided, for serum, AUIC24 h values of 37.1 for bacteriostasis, 46.3 for bactericidal activity and 119.6 for elimination of bacteria. These data may be used as a rational basis for setting dosing schedules which optimize clinical efficacy and minimize the opportunities for emergence of resistant organisms. PMID- 12081612 TI - Dose determination and confirmation of a long-acting formulation of ceftiofur (ceftiofur crystalline free acid) administered subcutaneously for the treatment of bovine respiratory disease. AB - The objective of this work was to determine and confirm an effective dose of ceftiofur crystalline free acid sterile oil suspension (CCFA-SS, 100 mg ceftiofur equivalents (CE)/mL], a long-acting single-administration ceftiofur formulation, for the treatment of the bacterial component of bovine respiratory disease (BRD). Study 1 was a dose determination study that used an intratracheal Mannheimia haemolytica (Pasteurella haemolytica) challenge model to evaluate single administration doses of CCFA-SS at 0.0, 1.1, 2.2, 3.3, 4.4 or 5.5 mg CE/kg body weight (BW) for the treatment of BRD. Data from this study were used to select doses for field testing in three multi-location clinical studies. In Study 2, the efficacy of a single administration dose of CCFA-SS at 4.4 mg CE/kg BW was compared with a negative control for the treatment of naturally occurring BRD in feedlot cattle. Treatments were administered when uniform clinical signs of BRD were present. Study 3 used a design similar to Study 2, and compared single administration doses of CCFA-SS at 3.0 or 4.4 mg CE/kg BW with the positive control tilmicosin (Micotil(R) 300 Injection, Elanco Animal Health) at 10 mg/kg BW. Study 4 compared the efficacy of single doses of CCFA-SS of 1.1-8.8 mg CE/kg BW with tilmicosin at 10 mg/kg BW. A total of 1176 cattle were included in these clinical studies. In Study 1, a dose of 4.55 mg CE/kg BW was determined to be effective. This was rounded to 4.4 mg CE/kg for field testing. In Study 2, a single dose of CCFA-SS at 4.4 mg CE/kg BW had a higher treatment success rate on day 14 (61%) than negative controls (26%, P < 0.01). However, in Study 3 this dose was judged to be at the beginning of an efficacious dose range for the treatment of BRD when compared with tilmicosin. In Study 4, day 28 treatment success rates were higher for CCFA-SS at 4.4-8.8 CE/kg BW than for tilmicosin (P=0.002) or the noneffective CCFA-SS dose of 1.1 mg CE/kg BW (P < 0.001). Based on decision criteria for Study 4, the effective dose was determined to be 4.4-5.5 mg CE/kg BW. These clinical studies demonstrated that a single dose of CCFA-SS (100 mg CE/mL) administered subcutaneously (s.c.) in the neck at 4.4-5.5 mg CE/kg BW is an effective treatment for BRD in feedlot cattle. However, this route of administration is no longer being considered for this formulation because of the ceftiofur residues that are present at the injection site for extended periods of time. PMID- 12081613 TI - The effect of intravenous administration of variable-dose flumazenil after fixed dose ketamine and midazolam in healthy cats. AB - The effects of intravenous administration of variable-dose flumazenil (0, 0.001, 0.005, 0.01, and 0.1 mg/kg) after ketamine (3 mg/kg) and midazolam (0.0 and 0.5 mg/kg) were studied in 18 healthy unmedicated cats from time of administration until full recovery. End-points were chosen to determine whether flumazenil shortened the recovery period and/or modified behaviors previously identified and attributed to midazolam. Overall, flumazenil administration had little effect on recovery or behaviors. One minute after flumazenil administration, all cats were recumbent but a greater proportion of cats which received the highest dose assumed sternal recumbency with head up than any other group. Although not significant, those cats that received the highest flumazenil dose also had shorter mean times for each of the initial recovery stages (lateral recumbency with head up, sternal recumbency with head up and walking with ataxia) than any of the other treatment groups that received midazolam. For complete recovery, flumazenil did decrease the proportion of the cats that was sedated, but did not shorten the time to walking without ataxia. Based on this study, the administration of flumazenil in veterinary practice, at the doses studied, to shorten and/or improve the recovery from ketamine and midazolam in healthy cats cannot be recommended. PMID- 12081614 TI - Bioavailability of transdermal methimazole in a pluronic lecithin organogel (PLO) in healthy cats. AB - The antithyroid drug methimazole is widely used for the medical management of feline hyperthyroidism. Recently, custom veterinary pharmacies have offered methimazole in a transdermal gel containing pluronic and lecithin (PLO), with anecdotal evidence of efficacy. The purpose of this study was to determine the bioavailability, relative to i.v. and oral routes of administration, of transdermal methimazole in a PLO gel in cats. Six healthy adult cats were assigned to receive 5 mg of methimazole by the i.v., oral, or transdermal routes, in a randomized triple crossover protocol with 1 week washout between doses. Blood samples were taken for high performance liquid chromatography (HPLC) determination of serum methimazole, at 0, 5, 15, 30, 60 min, and 2, 4, 6, 12 and 24 h after dosing. Methimazole absorption following transdermal administration was poor and variable, with only two of six cats achieving detectable serum methimazole concentrations at any time point following transdermal administration. Area under the concentration-time curve (AUC), maximum concentration (Cmax), and absolute bioavailability were all significantly lower for the transdermal route (0.39 +/- 0.63 microg h/mL, 0.05 +/- 0.09 microg/mL, and 11.4 +/- 18.7%, respectively) than for either i.v. (7.96 +/- 4.38 microg h/mL, 3.34 +/- 2.00 microg/mL, 100%) or oral routes (2.94 +/- 1.24 microg h/mL, 0.51 +/- 0.15 microg/mL, 40.4 +/- 8.1%). The results of this study indicate generally low to undetectable bioavailability of methimazole in a lecithin/pluronic gel given as a single transdermal dose to healthy cats, although one individual cat did achieve nearly 100% transdermal bioavailability relative to the oral route. PMID- 12081615 TI - Pharmacokinetics of phenylbutazone in beef steers. AB - Phenylbutazone was administered intravenously to a group of 11 beef steers at a dosage of 6 mg/kg of body weight. Whole plasma and protein-free plasma were analyzed for phenylbutazone residues. Pharmacokinetic parameters of total and free phenylbutazone in plasma were calculated using a noncompartmental method. In regards to whole plasma data, the mean volume of distribution at steady state (Vss), was 140 mL/kg body weight, with a mean (+/-SEM) terminal elimination half life (t1/2) of 34 +/- 9 h. The mean clearance was 3.2 mL/h/kg body weight. The Vss, as determined from the protein-free plasma fraction, was 54093 mL/kg body weight. This larger Vss of free phenylbutazone compared with total plasma phenylbutazone was attributed to a high degree of plasma protein binding, as well as the greater penetration of free phenylbutazone into tissues. The mean t1/2 of free phenylbutazone was 35 +/- 12 h. This similarity to the t1/2 estimated from total plasma phenylbutazone data is attributed to an equilibrium between free and plasma phenylbutazone during the terminal elimination phase. The pharmacokinetic parameters of free and total plasma phenylbutazone in beef steers are statistically similar to those previously reported for lactating dairy cows. PMID- 12081616 TI - Challenges associated with the evaluation of veterinary product bioequivalence: an AAVPT perspective. AB - The Generic Animal Drug Patent Term Restoration Act (GADPTRA) enacted in 1988 provided the same benefits to animal drug products that were granted to human generic products. It has been over 13 years since the GADPTRA was enacted, and veterinary drug sponsors and regulators have gained enormous insight and experience into some of the unique challenges associated with the determination of product bioequivalence for veterinary dosage forms. Moreover, advances in information and technology have opened both new issues that must be addressed and new mechanisms for demonstrating product bioequivalence. While many aspects of the existing Center for Veterinary Medicine Bioequivalence Guidance continue to provide invaluable guidance to the animal drug industry, there are also aspects of this guidance that are being called into question. Therefore, during the 2001 annual meeting of the American Academy of Veterinary Pharmacology and Therapeutics, participants were asked to address issues and concerns associated with the evaluation of veterinary product bioequivalence. This manuscript provides a summary of the concerns and discussions that transpired. PMID- 12081617 TI - Acute effects of escalating doses of amiodarone in isolated guinea pig hearts. AB - Cardiac effects of escalating concentrations of amiodarone were determined on isolated perfused guinea pig hearts (Langendorff preparations). Spontaneously beating hearts were instrumented for the measurement of RR, PQ, QRS, QT and QTc durations (from a bipolar electrogram), and dP/dtmax and dP/dtmin from an isovolumetric left ventricular pressure curve. Ten hearts were exposed to escalating concentrations of amiodarone (10-7, 10-6, 10-5 and 10-4 M) in dimethyl sulfoxide (DMSO)/Krebs-Henseleit or to DMSO/Krebs-Henseleit (vehicle). Measurements were collected during the last minute of a 15-min concentration. Means of all parameters were compared by ANOVA with repeated measures design. When compared with vehicle, amiodarone prolonged QT and QTc durations at concentrations >10-6 M. The apparent lengthening of RR, PQ and QRS at concentrations >10-6 M did not achieve statistical significance. Similarly, the apparent decreases in dP/dtmax and dP/dtmin at concentrations >10-6 M did not achieve statistical significance. The putative therapeutic concentration of amiodarone is between 2 and 4 x 10-6 M. In this study, at a concentration of 10-6 M, only RR and dP/dtmin tended to change, but they were not different from vehicle. Thus, amiodarone in this preparation has little potential for cardiac toxicity at therapeutic concentrations. PMID- 12081618 TI - The effects of body composition on the pharmacokinetics of subcutaneously injected ivermectin and moxidectin in pigs. AB - Macrocyclic lactones are characterized by their long persistence in animals because of their extensive distribution into fat. This study examined the influence of body condition on the disposition of ivermectin (IVM) and moxidectin (MXD) in blood and fat following subcutaneous (s.c.) drug administration. 'Fat' and 'thin' lines of pigs were established using two different diets. All animals were then injected with either MXD or IVM at 300 microg/kg and blood samples were taken at regular intervals until slaughter. Two IVM-treated animals from each diet group were slaughtered at either 3 days or 3 weeks posttreatment. Two MXD treated animals from each diet group were slaughtered at 3 days, 3, 6 or 9 weeks after treatment. Samples of backfat were taken from all animals at slaughter. Fluorescence HPLC was used to determine the concentrations of MXD or IVM in the plasma and fat samples. The plasma IVM concentration peaked more rapidly in the thin IVM treated pigs compared with the fat pigs. The concentration of IVM in backfat was significantly lower in the thin animals slaughtered 3 weeks after treatment. The MXD plasma concentration peaked within the first hour in both the thin and fat groups, but from 12 h posttreatment there was a higher MXD concentration in the plasma of the fat pigs resulting in MXD being detectable in these pigs for 28 days compared with only 17 days in the thin pigs. Despite this difference in plasma persistence no differences were seen in the MXD concentration of backfat between fat and thin animals. Body condition influenced the kinetic disposition of IVM and MXD following s.c. drug administration with both drugs being less persistent in thin compared with fat animals. PMID- 12081619 TI - The effect of tiletamine and zolazepam on isoflurane minimum alveolar concentration in goats. PMID- 12081620 TI - Disposition of oxytetracycline in pigs after i.m. administration of two long acting formulations. PMID- 12081621 TI - Effects of altered plasma alpha1-acid glycoprotein levels on pharmacokinetics of some basic antibiotics in pigs: simulation analysis. PMID- 12081622 TI - Solution structure of a cathelicidin-derived antimicrobial peptide, CRAMP as determined by NMR spectroscopy. AB - CRAMP was identified from a cDNA clone derived from mouse femoral marrow cells as a member of cathelicidin-derived antimicrobial peptides. This peptide shows potent antimicrobial activity against gram-positive and gram-negative bacteria but no hemolytic activity against human erythrocytes. CRAMP was known to cause rapid permeabilization of the inner membrane of Escherichia coli. In this study, the structure of CRAMP in TFE/H2O (1 : 1, v/v) solution was determined by CD and NMR spectroscopy. CD spectra showed that CRAMP adopts a mainly alpha-helical conformation in TFE/H2O solution, DPC micelles, SDS micelles and liposomes, whereas it has a random structure in aqueous solution. The tertiary structure of CRAMP in TFE/H2O (1 : 1, v/v), as determined by NMR spectroscopy, consists of two amphipathic alpha-helices from Leu4 to Lys10 and from Gly16 to Leu33. These two helices are connected by a flexible region from Gly11 to Gly16. Previous analysis of series of fragments composed of various portion of CRAMP revealed that an 18 residue fragment with the sequence from Gly16 to Leu33 was found to retain antibacterial activity. Therefore, the amphipathic alpha-helical region from Gly16 to Leu33 of CRAMP plays important roles in spanning the lipid bilayers as well as its antibiotic activity. Based on this structure, novel antibiotic peptides having strong antibiotic activity, with no hemolytic effect will be developed. PMID- 12081623 TI - Effect of asymmetric modification on the conformation of ascidiacyclamide analogs. AB - Ascidiacyclamide (ASC), cyclo(-Ile1-Oxz2-d-Val3-Thz4-)2 (Oxz=oxazoline and Thz=thiazole) has a C2-symmetric sequence, and the relationships between its conformation and symmetry have been studied. In a previous study, we performed asymmetric modifications in which an Ile residue was replaced by Gly, Leu or Phe to disturb the symmetry [Doi et al. (1999) Biopolymers49, 459-469]. In this study, the modifications were extended. The Ile1 residue was replaced by Gly, Ala, aminoisobutyric acid (Aib), Val, Leu, Phe or d-Ile, and the d-Val3 residue was replaced by Val. The structures of these analogs were analyzed by X-ray diffraction, 1H NMR and CD techniques. X-Ray diffraction analyses revealed that the [Ala1], [Aib1] and [Phe1]ASC analogs are folded, whereas [Val1]ASC has a square form. These structures are the first examples of folded structures for ASC analogs in the crystal state and are similar to the previously reported structures of [Gly1] and [Phe1]ASC in solution. The resonances of amide NH and Thz CH protons linearly shift with temperature changes; in particular, those of [Aib1], [d-Ile1] and [Val3]ASCs exhibited a large temperature dependence. DMSO titration caused nonlinear shifts of proton resonances for all analogs and largely affected [d-Ile1] and [Val3]ASCs. A similar tendency was observed upon the addition of acetone to peptide solutions. Regarding peptide concentration changes, amide NH and Thz CH protons of [Gly1]ASC showed a relatively large dependence. CD spectra of these analogs indicated approximately two patterns in MeCN solution, which were related to the crystal structures. However, all spectra showed a similar positive Cotton effect in TFE solution, except that of [Val3]ASC. In the cytotoxicity test using P388 cells, [Val1]ASC exhibited the strongest activity, whereas the epimers of ASC ([d-Ile1] and [Val3]ASCs), showed fairly moderate activities. PMID- 12081624 TI - Conformation and interaction of the cyclic cationic antimicrobial peptides in lipid bilayers. AB - To investigate the role of peptide-membrane interactions in the biological activity of cyclic cationic peptides, the conformations and interactions of four membrane-active antimicrobial peptides [based on Gramicidin S (GS)] were examined in neutral and negatively charged micelles and phospholipid vesicles, using CD and fluorescence spectroscopy and ultracentrifugation techniques. Moreover, the effects of these peptides on the release of entrapped fluorescent dye from unilamellar vesicles of phosphatidylcholine (PC) and phosphatidylethanolamine/phosphatidylglycerol (PE/PG) were studied. The cyclic peptides include GS10 [Cyclo(VKLdYP)2], GS12 [Cyclo(VKLKdYPKVKLdYP)], GS14 [Cyclo(VKLKVdYPLKVKLdYP)] and [d-Lys]4GS14 [Cyclo(VKLdKVdYPLKVKLdYP)] (underlined residues are d-amino acids), were different in their ring size, structure and amphipathicity, and covered a broad spectrum of hemolytic and antimicrobial activities. Interaction of the peptides with the zwitterionic PC and negatively charged PE/PG vesicles were distinct from each other. The hydrophobic interaction seems to be the dominant factor in the hemolytic activity of the peptides, as well as their interaction with the PC vesicles. A combination of electrostatic and hydrophobic interactions of the peptides induces aggregation and fusion in PE/PG vesicles with different propensities in the order: [d-Lys]4GS14 > GS14 > GS12 > GS10. GS10 and GS14 are apparently located in the deeper levels of the membrane interfaces and closer to the hydrophobic core of the bilayers, whereas GS12 and [d-Lys]4GS14 reside closer to the outer boundary of the interface. Because of differing modes of interaction of the cyclic cationic peptides with lipid bilayers, the mechanism of their biological activity (and its relation to peptide-lipid interaction) proved to be versatile and complex, and dependent on the biophysical properties of both the peptides and membranes. PMID- 12081625 TI - Design and characterization of a membrane permeable N-methyl amino acid containing peptide that inhibits Abeta1-40 fibrillogenesis. AB - Alzheimer's disease, Huntington's disease and prion diseases are part of a growing list of diseases associated with formation of beta-sheet containing fibrils. In a previous publication, we demonstrated that the self-association of the Alzheimer's beta-amyloid (Abeta) peptide is inhibited by peptides homologous to the central core domain of Abeta, but containing N-methyl amino acids at alternate positions. When these inhibitor peptides are arrayed in an extended, beta-strand conformation, the alternating position of N-methyl amino acids gives the peptide two distinct faces, one exhibiting a normal pattern of peptide backbone hydrogen bonds, but the other face having limited hydrogen-bonding capabilities due to the replacement of the amide protons by N-methyl groups. Here, we demonstrate, through two-dimensional NMR and circular dichroic spectroscopy, that a pentapeptide with two N-methyl amino acids, Abeta16-20m or Ac-K(Me)LV(Me)FF-NH2, does indeed have the intended structure of an extended beta strand. This structure is remarkably stable to changes in solvent conditions and resists denaturation by heating, changes in pH (from 2.5 to 10.5), and addition of denaturants such as urea and guanindine-HCl. We also show that this peptide, despite its hydrophobic composition, is highly water soluble, to concentrations > 30 mm, in contrast to the nonmethylated congener, Abeta16-20 (Ac-KLVFF-NH2). The striking water solubility, in combination with the hydrophobic composition of the peptide, suggested that the peptide might be able to pass spontaneously through cell membranes and model phospholipid bilayers such as unilamellar vesicles. Thus, we also demonstrate that this peptide is indeed able to pass spontaneously through both synthetic phospholipid bilayer vesicles and cell membranes. Characterization of the biophysical properties of the Abeta16-20m peptide may facilitate the application of this strategy to other systems as diverse as the HIV protease and chemokines, in which there is dimerization through beta-strand domains. PMID- 12081626 TI - A new B-chain mutant of insulin: comparison with the insulin crystal structure and role of sulfonate groups in the B-chain structure. AB - The solution structure of a new B-chain mutant of bovine insulin, in which the cysteines B7 and B19 are replaced by two serines, has been determined by circular dichroism, 2D-NMR and molecular modeling. This structure is compared with that of the oxidized B-chain of bovine insulin [Hawkins et al. (1995) Int. J. Peptide Protein Res.46, 424-433]. Circular dichroism spectroscopy showed in particular that a higher percentage of helical secondary structure for the B-chain mutant is estimated in trifluoroethanol solution in comparison with the oxidized B-chain. 2D-NMR experiments confirmed, among multiple conformations, that the B-chain mutant presents defined secondary structures such as a alpha-helix between residues B9 and B19, and a beta-turn between amino acids B20 and B23 in aqueous trifluoroethanol. The 3D structures, which are consistent with NMR data and were obtained using a simulated annealing protocol, showed that the tertiary structure of the B-chain mutant is better resolved and is more in agreement with the insulin crystal structure than the oxidized B-chain structure described by Hawkins et al. An explanation could be the presence of two sulfonate groups in the oxidized insulin B-chain. Either by their charges and/or their size, such chemical groups could play a destructuring effect and thus could favor peptide flexibility and conformational averaging. Thus, this study provides new insights on the folding of isolated B-chains. PMID- 12081628 TI - Aspirin and Reye's syndrome: is the response appropriate? PMID- 12081627 TI - Study of the binding environment of alpha-factor in its G protein-coupled receptor using fluorescence spectroscopy. AB - Mating in Saccharomyces cerevisiae is induced by the interaction of alpha-factor (W1H2W3L4Q5L6K7P8G9Q10P11M12Y13) with its cognate G protein-coupled receptor (Ste2p). Fifteen fluorescently labeled analogs of alpha-factor in which the 7 nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) group was placed at the alphaN-terminus and in side-chains at positions 1, 3, 4, 6, 7, 12 and 13 were synthesized and assayed for biological activity and receptor affinity. Eleven of the analogs retained 6 60% of the biological activity of the alpha-factor, as judged using a growth arrest assay. The binding affinities depended on the position of NBD attachment in the peptide and the distance of the tag from the backbone. Derivatization of the positions 3 and 7 side-chains with the NBD group resulted in analogs with affinities of 17-35% compared with that of alpha-factor. None of the other NBD containing agonists had sufficient receptor affinity or strong enough emission for fluorescence analysis. The position 3 and 7 analogs were investigated using fluorescence spectroscopy and collisional quenching by KI in the presence of Ste2p in yeast membranes. The results showed that the lambda max of NBD in the position 7 side-chain shifted markedly to the blue (510 nm) when separated by 4 or 6 bonds from the peptide backbone and that this probe was shielded from quenching by KI. In contrast, separation by 3, 5, 10 or more bonds resulted in lambda max ( approximately 540 nm) and collisional quenching constants consistent with increasing degrees of exposure. The NBD group in the position 3 side-chain was also found to be blue shifted (lambda max=520 nm) and shielded from solvent. These results indicate that the position 7 side-chain is likely interacting with a pocket formed by extracellular domains of Ste2p, whereas the side-chain of Trp3 is in a hydrophobic pocket possibly within the transmembrane region of the receptor. PMID- 12081629 TI - Indirect modulation of dopamine D2 receptors as potential pharmacotherapy for schizophrenia: III. Retinoids. AB - Present antipsychotic drugs, whose clinical activity correlates with direct binding to dopamine D2 or other receptors, alleviate some of the symptoms of schizophrenia, but not all and not completely in many patients. In continuation of our overview of potential novel antipsychotic pharmacotherapy that would be based upon indirect modulation of dopamine or other neurotransmitter functioning, we focus in this article on the postulated use of retinoid analogs as novel antipsychotic agents. Several lines of evidence can be viewed as implicating retinoid dysregulation in schizophrenia, either as a causative or contributory factor. It has been proposed that using retinoid analogs to alter the downstream expression of dopamine D2 receptors might represent a novel approach to the treatment of the disease or amelioration of symptoms when used either as monotherapy or as adjunct pharmacotherapy to dopamine D2 receptor antagonists. PMID- 12081630 TI - Potential novel targets for Alzheimer pharmacotherapy: I. Secretases. AB - The prevailing major theory of Alzheimer's disease (AD) is that insoluble amyloid beta-peptide (Abeta) found in the cerebral plaques characteristic of the disease is causative or is at least a contributing factor. According to this theory, inhibition of aberrant Abeta production should prevent or at least limit the extent of AD pathophysiology. As three 'secretase' enzymes (alpha, beta and gamma) catalyse the proteolytic cleavage of amyloid precursor protein (APP) (the precursor protein of Abeta), one or more secretases have become targets for potential novel AD pharmacotherapy. Secretase inhibitors have been designed and are in various stages of development. The clinical trials of these compounds will, if positive, result in drugs with dramatically better clinical efficacy or, if negative, will force a reassessment of the theory about the role of Abeta in AD. PMID- 12081631 TI - Co-phenylcaine as an alternative to Brompton's solution in rigid nasendoscopy: a pilot study. AB - BACKGROUND: Rigid nasendoscopy is a diagnostic aid that is frequently used in otorhinolaryngology clinics. Topical local anaesthetic and vasoconstriction is advised prior to the procedure. Options include both Brompton's solution and co phenylcaine. Brompton's solution contains 10% cocaine, a controlled drug that has potentially serious side-effects. Even deaths have been reported as a consequence of its use as a nasal preparatory agent. OBJECTIVE: To compare the efficacy of cocaine in the form of Brompton's solution versus co-phenylcaine as both a vasoconstrictor and a local anaesthetic in out-patient diagnostic rigid nasendoscopy. METHOD: Thirty-three patients awaiting diagnostic rigid nasendoscopy were recruited to the study. Each patient served as his own control. Co-phenylcaine was delivered as a metered dose of two sprays to one nasal passage and Brompton's solution was delivered on a cotton wool pledget placed in the other nasal passage with the aid of Tilly's forceps. An endoscopist who was unaware of which nasal passage was treated with which agent scored the vasoconstricion at endoscopy 10 min after drug application using a scoring sheet. The patients gave a numerical assessment of how painful the procedure was for each nasal passage. Mean scores were compared using Student's t-test. RESULTS: There was no difference between the two agents with regard to nasal analgesia and vasoconstriction during rigid nasal endoscopy. PMID- 12081632 TI - Bayesian population model of methotrexate to guide dosage adjustments for folate rescue in patients with breast cancer. AB - BACKGROUND: Methotrexate (MTX) infusions may induce severe side-effects, and alkaline hydration along with folinic acid rescue is a common way to reduce such toxic risks. The purpose of this study was to develop an adaptive rescue strategy based upon the early detection of patients with impaired MTX elimination. METHODS AND RESULTS: In this study, we propose a simple population-based Bayesian approach for predicting MTX plasma concentration from a limited number of samples, so as to adapt both duration and dosage of the rescue agent to be used next. Ten kinetic profiles obtained after 10 courses of MTX (1.5 g/m2) in seven patients with inflammatory breast cancer were used to establish the population pharmacokinetic parameters (Cl, 8.16 L/h; t1/2, 12.7 h). This population was next involved in the Bayesian estimation of MTX individual pharmacokinetic parameters from only two blood samples (T24 and T36 h), thus allowing one to forecast the elimination of this drug by predicting MTX levels at 48 h. According to the MTX concentrations predicted during the elimination phase, folinic acid rescue was then prolonged in patients likely to be overexposed. CONCLUSION: The Bayesian estimation presented in this study was an easy and convenient method to efficiently detect patients with impaired MTX elimination in routine clinical practice. This information enabled the introduction of strategies for minimizing the risk of severe drug toxicity. PMID- 12081633 TI - Prescribing indicators and their use by primary care groups to influence prescribing. AB - OBJECTIVE: To examine the prescribing incentive schemes used by primary care groups (PCGs); to determine the prescribing indicators used under these schemes; and to assess whether the schemes were seeking to improve the quality of prescribing as well as controlling prescribing costs. DESIGN: Cross-sectional survey. SETTING: A total of 145 PCGs in the London and South-East NHS regions. PARTICIPANTS: Prescribing advisers in each PCG. METHODS: Descriptions of the prescribing indicators monitored by each PCG were obtained from a questionnaire survey of PCGs at the end of the 1999-2000 financial year. Financial information on prescribing and details about the implementation of prescribing incentive schemes for this period became available 6 months later and were obtained by a further questionnaire, follow-up telephone and E-mail surveys. OUTCOME MEASURES: Prescribing indicators, prescribing budgets and spend. RESULTS: One hundred and twenty-one out of 145 (83%) PCGs replied to the questionnaires about prescribing indicators and 129 out of 145 (89%) replied with details about their prescribing costs. The most frequently monitored prescribing indicator was generic prescribing, used by 106 out of 121 (88%) PCGs. The most frequently used clinical areas for prescribing indicators were antibiotics (76% of PCGs), gastro intestinal prescribing (68%), non-steroidal anti-inflammatories (37%) and cardiovascular prescribing (32%). Seventy-six (63%) schemes also used non prescribing analysis & cost (PACT) based data for their incentive schemes such as information from prescribing audits and reviews of repeat prescribing protocols. Only 33 (23%) had reached agreement with their practices enabling all prescribing indicator information to be disseminated on a named basis to allow practices to examine each others' prescribing data. CONCLUSIONS: Prescribing incentive schemes usually include targets for improvements in prescribing quality as well as cost. PACT-based data were used for cost control and quality improvement but non-PACT data were almost entirely used to promote prescribing quality improvements. The validity of non-PACT data was questioned as was the choice of some indicators that appeared to have been selected without full consideration of current expert opinion. Further work is needed on which indicators are most likely to act as catalysts to prescribing change. PMID- 12081634 TI - Comparison of ceftazidime degradation in glass bottles and plastic bags under various conditions. AB - OBJECTIVE: To study the effect of type of container on ceftazidime stability in intravenous solutions. METHODS: One hundred millilitre polypropylene (PP) and polyvinyl chloride (PVC) bags and 100-mL glass bottles were filled with 5% dextrose or 0.9% sodium chloride solutions containing ceftazidime (Fortumset) at 40 mg/mL. Three containers of each solution were stored at 20 and 35 degrees C. One millilitre samples were drawn from each container at 0 and 20 h and assayed. Pyridine concentrations, the main degradation product of ceftazidime, were determined by high-pressure liquid chromatography. RESULTS: Pyridine levels increased during storage and were higher in PVC and PP bags than in glass bottles in both diluents. Solutions stored in PP bags showed better stability than in PVC bags. CONCLUSION: This study shows that ceftazidime undergoes slower degradation in PP than PVC containers although the difference is small. Glass bottles seems to be the better container for storing ceftazidime solutions, whatever storage temperature and diluent used. PMID- 12081635 TI - The single nucleotide polymorphism T1128C in the signal peptide of neuropeptide Y (NPY) was not identified in a Korean population. AB - OBJECTIVE: A single nucleotide polymorphism (SNP) T1128C, which causes an amino acid change from leucine(7) to proline(7) in the signal peptide of neuropeptide Y (NPY), has been found to be associated with multiple clinical parameters or disease. However, this SNP was not identified in the Japanese population. We asked if this is also true in the Korean population. METHODS: Genotyping was conducted by pyrosequencing, a newly developed real-time high-throughput SNP scoring technique. RESULTS: All 242 Korean subjects showed T1128/T1128 genotype. CONCLUSIONS: The T1128C SNP does not appear to exist in this Korean population. PMID- 12081636 TI - Variation in pharmacy prescription refill adherence measures by type of oral antihyperglycaemic drug therapy in seniors in Nova Scotia, Canada. AB - OBJECTIVE: To examine the association between pharmacy prescription refill adherence by type of oral antihyperglycaemic medications used in seniors in Nova Scotia, Canada. RESEARCH DESIGN AND METHODS: Pharmacy and health care utilization data from April 1993 to March 1996 for Nova Scotia Seniors' Pharmacare beneficiaries treated with 1st and 2nd generation sulphonylureas and biguanides was analysed. Refill adherence was quantified by two proportions: number of days beneficiaries had a medication surplus compared with the total period of observation and gaps in treatment compared with the total period of observation. Analysis examined association of type of oral antihyperglycaemic agent and dosing on refill adherence, after adjustment for age, gender and hospital use. RESULTS: A total of 3358 beneficiaries met the study criteria. The mean refill adherence rate [continuous multiple-interval measure of medication availability (CMA)] was 86 +/- 0.4% SE and continuous measure of medication gaps (CMG) was 16 +/- 0.4% SE. Use of biguanides was associated with lower odds of having a medication surplus. The use of 2nd generation sulphonylureas and biguanides, and use of agents with a dosage frequency of more than one dose per day was associated with medication gaps. CONCLUSIONS: Many beneficiaries taking antihyperglycaemic agents adhered well to prescribed therapy. The proportion of days not covered by medications averaged 16%. Beneficiaries taking medications once a day were more likely to have good refill adherence. Further work is needed to compare prescription refill adherence rates with other adherence measures and clinical outcomes. These methods are useful for establishing baseline adherence, monitoring the success of programmes designed to improve adherence, and determining cost-effectiveness of drug regimens. PMID- 12081637 TI - The effects on GP prescribing of joining a commissioning group. AB - BACKGROUND: It is not known to what extent general practitioners (GP) can change their prescribing upon joining a commissioning group and what features of a commissioning group may promote prescribing change. The opportunity to study potential prescribing change arose with the formation of a limited number of Primary Care Commissioning Groups (PCCGs), a precursor of Primary Care Groups (PCGs) and Primary Care Trusts (PCTs). METHODS: This was a controlled study of general practice prescribing costs. All practices (n=24) within one inner city PCCG were compared with matched controls that were not part of a PCCG. Cross sectional survey data was collected from the PCCG practices to determine possible reasons for prescribing change. RESULTS: The total annual prescribing cost rose by 4.0% in the PCCG practices and by 6.9% in controls (P=0.01). Significant cost containment was found for gastrointestinal prescribing (P=0.03), attributable to differences in the cost of proton pump inhibitors (PPIs) which fell by 0.7% in the PCCG but rose by 7.3% in controls (P=0.03). Total relative savings in the PCCG practices amounted to around pound 220,000. General practitioners making the greater savings in PPI costs within the PCCG, were more likely to report being influenced by information from the prescribing adviser. CONCLUSION: General practice prescribing costs were contained to a greater degree in practices participating in the PCCG. The differences in gastrointestinal prescribing were most marked for PPIs which were specifically targeted by the prescribing adviser. The GPs themselves attributed their own prescribing change to information provided by the prescribing adviser. Other factors operating within the PCCG may also have influenced prescribing such as a more locally based management system, different financial incentives and a greater degree of co-operative working amongst GPs. PMID- 12081638 TI - Paeoniae Radix, a traditional Chinese medicine, and CYP2C9 activity. AB - Paeoniae Radix (PR) is a commonly used traditional Chinese medicine. A slight effect of PR on the pharmacokinetics of phenytoin that is mainly metabolised by CYP2C9 has been reported. The aim of this pilot study was to clarify if PR has an effect on losartan oxidation used as a measure of CYP2C9 activity. Three healthy volunteers received a single oral dose of losartan before and after PR treatment. Losartan and E-3174, an active metabolite of losartan, were analysed in 8 hour urine. PR did not seem to have an effect on CYP2C9 activity when the losartan/E 3174 ratios were compared before and after PR treatment (P = 0.56) although a larger study would need to be undertaken to confirm this finding. PMID- 12081640 TI - Calcineurin phosphatase in signal transduction: lessons from fission yeast. AB - Calcineurin (protein phosphatase 2B), the only serine/threonine phosphatase under the control of Ca2+/calmodulin, is an important mediator in signal transmission, connecting the Ca2+-dependent signalling to a wide variety of cellular responses. Furthermore, calcineurin is specifically inhibited by the immunosuppressant drugs cyclosporin A and tacrolimus (FK506), and these drugs have been a powerful tool for identifying many of the roles of calcineurin. Calcineurin is enriched in the neural tissues, and also distributes broadly in other tissues. The structure of the protein is highly conserved from yeast to man. The combined use of powerful genetics and of specific calcineurin inhibitors in fission yeast Schizosaccharomyces pombe (S. pombe) identified new components of the calcineurin pathway, and defined new roles of calcineurin in the regulation of the many cellular processes. Recent data has revealed functional interactions in which calcineurin phosphatase is involved, such as the cross-talk between the Pmk1 MAP kinase signalling, or the PI signalling. Calcineurin also participates in membrane traffic and cytokinesis of fission yeast through its functional connection with members of the small GTPase Rab/Ypt family, and Type II myosin, respectively. These findings highlight the potential of fission yeast genetic studies to elucidate conserved elements of signal transduction cascades. PMID- 12081641 TI - SsrA-mediated protein tagging in the presence of miscoding drugs and its physiological role in Escherichia coli. AB - BACKGROUND: We have shown recently that read-through of a normal stop codon by a suppressor tRNA in specific genes possessing a Rho-independent terminator leads to SsrA-mediated tagging of extended proteins in Escherichia coli cells. Miscoding antibiotics such as kanamycin and streptomycin reduce translational fidelity by binding to the 30S ribosomal subunit. The aim of the present study was to address how miscoding antibiotics affect the read-through of stop codons and SsrA-mediated protein tagging. RESULTS: Miscoding antibiotics caused translational read-through of stop codons when added to the culture medium at sublethal concentrations. Under the same conditions, the drugs enhanced SsrA mediated tagging of bulk cellular proteins, as observed in cells carrying an ochre suppressor tRNA. Translational read-through products generated from the crp gene in the presence of the antibiotics was efficiently tagged by the SsrA system, presumably because the ribosome reached the 3' end of the mRNA defined by the terminator hairpin. The SsrA-defective cells were more sensitive to the miscoding antibiotics compared to the wild-type cells. CONCLUSION: We conclude that the SsrA system contributes to the survival of cells by dealing with translational errors in the presence of low concentrations of miscoding antibiotics. PMID- 12081642 TI - Over-expression of human DNA polymerase lambda in E. coli and characterization of the recombinant enzyme. AB - BACKGROUND: DNA polymerase lambda (Pol lambda) was recently identified as a new member of the family X of DNA polymerases in eukaryotic cells. Pol lambda contains a nuclear localization signal (NLS), a BRCA1-C terminal (BRCT) domain, a proline-rich region, helix-hairpin-helix (HhH) and pol X motifs. Since the amino acid sequence for Pol lambda shares a high degree of homology to Pol beta, Pol lambda is considered to have a similar enzymatic nature to Pol beta. RESULTS: Recombinant human Pol lambda was shown to possess template-directed DNA polymerase activity in its monomeric form. Pol lambda required either Mn2+ or Mg2+ as a metal co-factor to catalyse this activity, and optimal activity was detected at pH 8.5-9.0. Pol lambda was insensitive to aphidicolin, but was sensitive to dideoxynucleoside triphosphates or N-ethylmaleimide. By constructing the truncated Pol lambda, the proline rich region was shown to act in a suppression of its polymerization activity. A chimeric enzyme comprised of the Pol lambda N-terminal region and Pol beta also showed a reduced Pol beta activity. Proliferating cell nuclear antigen (PCNA) directly interacts with Pol lambda through its Pol beta like region in vitro. CONCLUSIONS: Pol lambda possesses similar enzymatic nature to Pol beta; requirements of cations and optimal conditions for pH and NaCl concentration, aside from sensitivity to N ethylmaleimide and template preference. The proline rich region of Pol lambda functions as a suppressor domain for its polymerization activity (SDPA). Pol lambda interacts directly with PCNA through its Pol beta like region. The functional consequence of this interaction is the negative regulation of Pol lambda activity. PMID- 12081643 TI - The Cpx stress response system of Escherichia coli senses plasma membrane proteins and controls HtpX, a membrane protease with a cytosolic active site. AB - BACKGROUND: The abnormal accumulation of misfolded proteins outside the plasma (cytoplasmic or inner) membrane up-regulates the synthesis of a class of envelope localized catalysts of protein folding and degradation. The pathway for this transmembrane signalling is mediated by the CpxR-CpxA two-component phospho-relay mechanism. RESULTS: We now show that an abnormality in the plasma membrane proteins, due either to the impairment of FtsH, a protease acting against integral membrane proteins, or to the overproduction of a substrate membrane protein of FtsH, activates this stress response pathway. Under such conditions, the cpxR gene function becomes essential for cell growth. We further show that the expression of a putative protease, HtpX, in the plasma membrane, is under the control of CpxR. Synthetic growth inhibition was observed when the ftsH and htpX disruption mutations had been combined, suggesting that these gene products have some complementary or overlapping proteolytic functions. Topology analyses indicated that the metalloproteinase active site of HtpX is located on the cytosolic side of the membrane. CONCLUSIONS: Taken together, these results suggest that the Cpx "extracytoplasmic" stress response system controls the quality of the plasma membrane, even on its cytoplasmic side. PMID- 12081644 TI - Fission yeast chk1 mutants show distinct responses to different types of DNA damaging treatments. AB - BACKGROUND: Chk1 kinase is activated by phosphorylation at serine-345 by Rad3 checkpoint kinase and is required for DNA damage checkpoint in late S and G2 phase of S. pombe cell cycle. We studied the ability of two chk1 mutants, chk1-1 and chk1-2, to undergo phosphorylation and to delay cell cycle progression in response to different types of DNA lesions. RESULTS: Both the Chk1-1 and Chk1-2 mutant proteins are phosphorylated to various extents when DNA is damaged in early G2 phase of cell cycle by either UV irradiation or gamma irradiation. However, chk1-2 mutant does not delay cell cycle progression in a dose dependent manner specifically upon gamma irradiations. This defect is not associated with an important loss of survival. Furthermore, both chk1 mutants survive to Camptothecin treatment despite undetectable Chk1-1 or Chk1-2 phosphorylated forms. We show that both mutant proteins are not phosphorylated in cds1 devoid cells treated with ribonucleotide reductase inhibitor hydroxyurea or when the replisome is affected by a thermosensitive mutation in DNA polymerase delta. This inability is associated with the loss of checkpoint function. We found that an increased level of Crb2/Rhp9 protein specifically complements the defect of the chk1-1 mutant allowing Chk1-1 phosphorylation upon treatment with hydroxyurea of dcds1 cells. CONCLUSIONS: Mutants chk1-1 and chk1-2 behave differently according to the type of lesion generated on DNA. PMID- 12081646 TI - Characterization of the chicken L-Maf, MafB and c-Maf in crystallin gene regulation and lens differentiation. AB - BACKGROUND: Members of the Maf family, including L-Maf, MafB and c-Maf, are "basic region/leucine zipper" (bZIP) transcription factors. Maf proteins contain a highly conserved acidic transactivation domain (AD), and a bZIP region that mediates DNA-binding activity. The hinge region between AD and bZIP varies considerably in length between different proteins. Recent studies reveal that L Maf, c-Maf and MafB play key roles in vertebrate lens development. RESULTS: We investigated the transactivation activity of individual factors in culture cells to analyse their specific functions. In transient transfection assays with a reporter gene containing Maf responsive elements, MafB and c-Maf activated higher levels of the reporter gene than L-Maf. However, L-Maf transactivated the alphaA crystallin promoter as effectively as MafB and c-Maf, and induced the expression of the endogenous delta-crystallin gene more efficiently than the other two proteins. Domain-swapping experiments reveal that the bZIP region of MafB takes part in strong transcriptional activity, while the acidic and hinge regions (AH) of c-Maf collectively serve as a strong transactivation domain. The AH region of L-Maf (but not c-Maf) conferred transactivation activity to induce delta crystallin gene expression. CONCLUSIONS: These results suggest that despite their similar DNA binding properties, L-Maf, MafB and c-Maf regulate different sets of target genes by complex interactions with multiple factors that recognize cis elements in promoters. The AH region of L-Maf has a distinct role in inducing endogenous delta-crystallin gene. PMID- 12081645 TI - The CLN3/SWI6/CLN2 pathway and SNF1 act sequentially to regulate meiotic initiation in Saccharomyces cerevisiae. AB - BACKGROUND: IME1, which is required for the initiation of meiosis, is regulated by Cln3:Cdc28 kinase, which activates the G1-to-S transition, and Snf1 kinase, which mediates glucose repression. Here we examine the pathway by which Cln3:Cdc28p represses IME1 and the relationship between Cln3:Cdc28p and Snf1p in this regulation. RESULTS: When wild-type yeast cease growth, they express IME1 to moderate levels, intermediate between the low levels expressed during growth and the high levels expressed during sporulation. Moderate IME1 expression occurred in cln3Delta, cln1Delta cln2Delta, cdc28-4 and swi6Delta mutants, even during growth. These mutants also induced IME1 expression more rapidly than the wild type. CLN3 required SWI6 and CLN2 to repress IME1 and IME2, but CLN1 was much less active than CLN2 in this repression. The phenotype of the cln3Delta snf1Delta double mutant indicated that Cln3:Cdc28p regulates IME1 independently of SNF1. CONCLUSION: Entry into meiosis involves two independent but sequential controls, which regulate IME1 via a three position switch: (i) during growth IME1 is repressed by the CLN3/SWI6/CLN2 pathway, (ii) once growth ceases, this repression is released and IME1 is expressed at moderate levels, and (iii) subsequently, nutritional conditions that activate Snf1p allow high IME1 expression. PMID- 12081647 TI - The SH3, HOOK and guanylate kinase-like domains of hDLG are important for its cytoplasmic localization. AB - BACKGROUND: hDLG, the human homologue of the Drosophila tumour suppressor dlg, functions as a scaffolding protein that facilitates the transmission of diverse downstream signals. hDLG possesses multiple protein-binding domains, including three PDZ domains, an SH3 domain, a HOOK domain and a guanylate kinase-like (GK) domain. RESULTS: We studied the significance of the PDZ, SH3, HOOK and GK domains in the cytoplasmic localization of hDLG. We found that mutation of the SH3 or GK domain, but not the PDZ domain, resulted in a re-localization of hDLG to the nucleus. Furthermore, hDLG was found to possess a potential nuclear localization signal in the HOOK domain. CONCLUSION: These results suggest that the SH3, HOOK and GK domains of hDLG are important for its cytoplasmic localization. PMID- 12081648 TI - Sexually dimorphic expression of Dax-1 in the adrenal cortex. AB - BACKGROUND: The DAX-1 (also known as AHC) gene encodes an unusual member of the nuclear receptor superfamily, and the gene product plays a pivotal role during gonadal and adrenal differentiation. Mutations of the human DAX-1 gene cause X linked adrenal hypoplasia congenita associated with hypogonadotropic hypogonadism. However, the molecular mechanisms underlying the transcriptional regulation of the gene are not well understood. RESULTS: Sexually dimorphic expression of Dax-1 (NR0B1) in the adrenal cortex was observed by RT-PCR, Western blotting and immunohistochemistry. The differential expression was abolished by gonadectomy and was restored again by sex steroid replacement. Our results suggested that the Dax-1 gene transcription is suppressed by androgens and androgen receptor (AR/NR3C4). Dax-1 gene transcription is regulated by Ad4BP/SF-1 (NR5A1), therefore we investigated the functional correlation between A4BP/SF-1 and AR. Interestingly, AR down-regulated the Dax-1 gene transcription mediated by Ad4BP/SF-1 in the presence of the ligand. DNA binding by AR was not essential for the suppressive action, however the suppression seemed to be dependent on the promoter contexts. An interaction between Ad4BP/SF-1 and AR was detected in the presence of the ligand for AR. CONCLUSION: The present study revealed that the expression of Dax-1 in the adrenal cortex is regulated by androgen and the receptor. Interestingly, AR acts as a suppressor in the presence of the ligand through interaction with Ad4BP/SF-1. PMID- 12081649 TI - A role of the C-terminus of aquaporin 4 in its membrane expression in cultured astrocytes. AB - BACKGROUND: Aquaporin 4 (AQP4) is a predominant water channel protein in mammalian brains, which is localized in the astrocyte plasma membrane. Membrane targeting of AQP4 is essential to perform its function. The mechanism(s) of membrane targeting is not clear in astrocytes. RESULTS: We investigated the role of the C-terminus of AQP4 (short isoform) in its membrane targeting by an expression study of C-terminal mutants of AQP4 in cultured astrocytes. The deletion of 26 C-terminal residues of AQP4 (AQP4[Delta276-301aa]) results in the intracellular localization of the protein. However, smaller deletions than 21 C terminal residues did not alter its plasma membrane localization. These results suggest that C-terminal residues between Val(276) and Ile(280) play an important role in the expression of AQP4 in the plasma membrane. However, the plasma membrane localization of the AQP4(A(276)AAAA(280)) mutant (alanine substitution of Val(276)-Ile(280) of AQP4) suggests that another signal for membrane targeting exists in the C-terminus of AQP4. The deletion or point mutations of the PDZ binding motif of the AQP4(A(276)AAAA(280)) mutant resulted in the intracellular localization of the proteins. These results suggest that the PDZ binding motif may also be involved in the membrane targeting of AQP4. CONCLUSIONS: We found that the C-terminal sequence of AQP4 contains two important signals for membrane expression of AQP4 in cultured astrocytes. One is a hydrophobic domain and the other is a PDZ binding motif that exists in the C-terminus. PMID- 12081650 TI - Vacuolar membrane dynamics revealed by GFP-AtVam3 fusion protein. AB - BACKGROUND: The plant vacuole is a multifunctional organelle that has various physiological functions. The vacuole dynamically changes its function and shape, dependent on developmental and physiological conditions. Our current understanding of the dynamic processes of vacuolar morphogenesis has suffered from the lack of a marker for observing these processes in living cells. RESULTS: We have developed transgenic Arabidopsis thaliana expressing a vacuolar syntaxin related molecule (AtVam3/SYP22) fused with green fluorescent protein (GFP). Observations using confocal laser scanning microscopy demonstrated that the plant vacuole contained a dynamic membrane system that underwent a complex architectural remodelling. Three-dimensional reconstitution and time-lapse analysis of GFP-fluorescence images revealed that cylindrical and sheet-like structures were present in the vacuolar lumen and were moving dynamically. The movement, but not the structure itself, was abolished by cytochalasin D, an inhibitor of actin polymerization. This moving structure, which sometimes penetrated through the vacuolar lumen, possessed a dynamic membrane architecture similar to the previously recognized "transvacuolar strand." CONCLUSION: We propose two possible models for the formation of the vacuolar lumenal structure. Membrane structures including protruding tubules and reticular networks have recently been recognized in many other organelles, and may be actively involved in intra- and/or inter-organelle signalling. PMID- 12081651 TI - The contribution of the intrinsic excitability of vestibular nucleus neurons to recovery from vestibular damage. AB - Damage to the peripheral vestibular system results in a syndrome of ocular motor and postural abnormalities that partially and gradually abate over time in a process known as 'vestibular compensation'. The first, rapid, phase of compensation has been associated with a recovery of spontaneous resting activity in the ipsilateral vestibular nucleus complex (VNC), as a consequence of neuronal and synaptic plasticity. Increasing evidence suggests that normal VNC neurons in labyrinthine-intact animals, as well as ipsilateral VNC neurons following unilateral vestibular deafferentation (UVD), rely to some extent on intrinsic pacemaker activity provided by voltage-dependent conductances for their resting activity. Modification of this intrinsic pacemaker activity may underlie the recovery of resting activity that occurs in ipsilateral VNC neurons following UVD. This review summarizes and critically evaluates the 'intrinsic mechanism hypothesis', identifying discrepancies amongst the current evidence and suggesting experiments that may test it further. PMID- 12081652 TI - Modulation by adenosine of both muscarinic M1-facilitation and M2-inhibition of [3H]-acetylcholine release from the rat motor nerve terminals. AB - The crosstalk between adenosine and muscarinic autoreceptors regulating evoked [3H]-acetylcholine ([3H]-ACh) release was investigated on rat phrenic nerve hemidiaphragm preparations. Motor nerve terminals possess facilitatory M1 and inhibitory M2 autoreceptors that can be activated by McN-A-343 (1-30 microm) and oxotremorine (0.3-100 microm), respectively. The muscarinic receptor antagonist, dicyclomine (3 nm-10 microm), caused a biphasic (inhibitory/facilitatory) effect, indicating that M1-facilitation prevails during 5 Hz stimulation trains. Concomitant activation of AF-DX 116-sensitive M2 receptors was partially attenuated, as pretreatment with M1 antagonists, muscarinic toxin 7 (MT-7, 0.1 nm) and pirenzepine (1 nm), significantly enhanced inhibition by oxotremorine. Activation of A2A-adenosine receptors with CGS 21680C (2 nm) (i) potentiated oxotremorine inhibition, and (ii) shifted McN-A-343-induced facilitation into a small inhibitory effect. Conversely, the A1-receptor agonist, R-N6 phenylisopropyl adenosine (R-PIA, 100 nm), attenuated the inhibitory effect of oxotremorine, without changing facilitation by McN-A-343. Synergism between A2A and M2 receptors is regulated by a reciprocal interaction with facilitatory M1 receptors, which may be prevented by pirenzepine (1 nm). During 50 Hz-bursts, facilitation (M1) of [3H]-ACh release by McN-A-343 disappeared, while the inhibitory (M2) effect of oxotremorine became predominant. This muscarinic shift results from the interplay with A2A receptors, as it was precluded by the selective A2A receptor antagonist, ZM 241385 (10 nm). In conclusion, when the muscarinic M1 positive feedback loop is fully operative, negative regulation of ACh release is mediated by adenosine A1 receptors. During high frequency bursts, tonic activation of A2A receptors promotes M2 autoinhibition by braking the M1 receptor operated counteraction. PMID- 12081653 TI - Intrinsic optical signals in the dorsal horn of rat spinal cord slices elicited by brief repetitive stimulation. AB - With repetitive electrical stimulation of the dorsal root (20 Hz for 1 s at C fibre strength), intrinsic optical signals (IOSs), measured as changes in light transmittance, were recorded in the superficial dorsal horn of rat spinal cord slices using a photodiode array imaging device. The mechanism underlying the induction of IOSs was investigated. IOSs elicited by brief repetitive stimulation persisted for 1-2 min and were decreased by reducing external Cl- concentration or by cation-chloride cotransport inhibitors. Furosemide was most effective whilst bumetanide was least effective among the inhibitors tested. A 1-min elevation of external K+ concentration evoked IOSs in the dorsal horn in the absence of stimulation, and K+-induced IOSs were inhibited by furosemide. These results suggest that the uptake of excess K+ via the furosemide-sensitive, cation chloride cotransporters underlies the induction of the IOSs. One-minute exposure to hypotonic solutions, which would cause cell swelling, induced IOSs in the superficial dorsal horn. Whilst osmotic-induced IOSs were not affected by furosemide, they were inhibited by HgCl2 in a 2-mercaptoethanol-sensitive manner. The stimulation-induced IOSs were similarly depressed by HgCl2. In contrast, voltage-sensitive dye signals and field potentials, evoked by single electrical stimuli, were significantly less affected by HgCl2. These results suggest that there is a specialized water transport pathway in the superficial dorsal horn, and that IOSs elicited by brief repetitive activation of C-fibres are attributable to cell swelling caused by water influx through this pathway, as an osmotic gradient is established by the uptake of K+ via the furosemide-sensitive cotransporters. PMID- 12081654 TI - Expression and dendritic mRNA localization of GABAC receptor rho1 and rho2 subunits in developing rat brain and spinal cord. AB - The cellular distribution of GABAC receptor rho1 and rho2 subunits in the rat central nervous system remains controversial. We investigated how these subunits were distributed in cerebellum, hippocampus and spinal cord at postnatal day 1, 7 or in adult life. We found that in the adult cerebellum rho1 and rho2 mRNAs were expressed in Purkinje cells and basket-like cells only. In the hippocampus both subunits were expressed throughout the CA1 pyramidal layer, dentate gyrus and scattered interneurons with maximum staining intensity at P7. In the adult hippocampus in situ staining was predominantly found on interneurons. GABAC antibody labelling in P7 and adult hippocampus was largely overlapping with the in situ staining. Western blot analysis showed GABAC receptor in retina, ovary and testis. In the spinal cord the rho2 signal was consistently stronger than rho1 with overlapping expression patterns. At P1, the most intensely labelled cells were the motoneurons while on P7 and adult sections, interneurons and motoneurons were likewise labelled. On spinal neurons both rho1 and rho2 mRNAs showed somatodendritic localization, extending out for >100 microm with punctate appearance especially in adult cells. A similar spinal distribution pattern was provided with polyclonal antibody labelling, suggesting close correspondence between mRNA and protein compartmentalization. Electrophysiological experiments indicated that P1 spinal motoneurons did possess functional GABAC receptors even though GABAC receptors played little role in evoked synaptic transmission. Our results suggest a pattern of rho1 and rho2 subunit distribution more widespread than hitherto suspected with strong developmental regulation of subunit occurrence. PMID- 12081655 TI - Reminder effects - reconsolidation or retrieval deficit? Pharmacological dissection with protein synthesis inhibitors following reminder for a passive avoidance task in young chicks. AB - It is generally accepted that memory formation involves an irreversible passage via labile phases to the stable form of 'long-term memory' impervious to amnestic agents such as protein synthesis inhibitors. However, recent experiments demonstrate that reactivation of memory by way of a reminder renders it labile to such inhibitors, suggesting that such retrieval is followed by a so-called reconsolidation process similar or identical in its cellular and molecular correlates to that occurring during the initial consolidation. We compared the effects of the protein synthesis inhibitor anisomycin and the glycoprotein synthesis inhibitor 2-deoxygalactose on the temporal dynamics and pharmacological sensitivity of initial consolidation and memory expression following a reminder in a one-trial passive-avoidance task in day-old chicks. This comparison revealed three differences between the action of the inhibitors on newly formed compared with reactivated memory. First, the recall deficit after the reminder was temporary, whilst the amnesia following inhibitor treatment during training was stable. Second, the sensitive period for the effect of anisomycin was shorter in the reminder than in the training situation. Third, the effective dose for either inhibitor for reminder-associated amnesia was several times lower than for amnesia developing after training. Thus though like initial consolidation, memory expression at delayed periods following reminder depends on protein and glycoprotein synthesis, the differences between the temporal and pharmacological dynamics in the two situations point to the distinct character of the molecular processes involved in postreminder effects. PMID- 12081656 TI - Expression and distribution of metabotropic GABA receptor subtypes GABABR1 and GABABR2 during rat neocortical development. AB - To understand the possible contribution of metabotropic gamma-aminobutyric acid receptors (GABABR) in cortical development, we investigated the expression pattern and the cellular and subcellular localization of the GABABR1 and GABABR2 subtypes in the rat neocortex from embryonic day 14 (E14) to adulthood. At the light microscopic level, both GABABR1 and GABABR2 were detected as early as E14. During prenatal development, both subtypes were expressed highly in the cortical plate. Using double immunofluorescence, GABABR1 colocalized with GABABR2 in neurons of the marginal zone and subplate, indicating that these proteins are coexpressed and could be forming functional GABABRs during prenatal development in vivo. In contrast, only GABABR1 but not GABABR2 was detected in the tangentially migratory cells in the lower intermediate zone. During postnatal development, immunoreactivity for GABABR1 and GABABR2 was distributed mainly in pyramidal cells. Discrete GABABR1-immunopositive cell bodies of interneurons were present throughout the neocortex. In addition, GABABR1 but not GABABR2 was found in identified Cajal-Retzius cells in layer I. At the electron microscopic level, immunoreactivity for GABABR1 and GABABR2 was found in dendritic spines and dendritic shafts at extrasynaptic and perisynaptic sites throughout postnatal development. We further demonstrated the presynaptic localization of GABABR1 and GABABR2, as well as the association of the receptors with asymmetrical synaptic junctions. These results indicate potentially important roles for the GABABRs in the regulation of migratory processes during corticogenesis and in the modulation of synaptic transmission during early development of cortical circuitry. PMID- 12081657 TI - The age-related increase in IL-1 type I receptor in rat hippocampus is coupled with an increase in caspase-3 activation. AB - Evidence from several studies indicates that expression of interleukin-1beta (IL 1beta) and IL-1 type I receptor is particularly high in hippocampus, and it has recently been shown that the concentration of IL-1beta is increased in the hippocampus of the aged rat. Here we report that this increase is coupled with an increase in expression of IL-1 type I receptor and increased activity of IL-1 receptor-associated kinase. The evidence presented indicates that the age-related increase in activity of the mitogen-activated protein kinases, Jun N-terminal kinase (JNK) and p38, was accompanied by enhanced caspase-3 activity. Analysis of colocalization of activated caspase-3 with activated p38 (p-p38) suggested that p p38 was necessary for activation of caspase-3; while in vitro analysis indicated that the IL-1beta-induced increase in caspase-3 activity was abrogated by the p38 inhibitor, SB203580. The IL-1beta-induced increase in caspase-3 activity in vitro was also abrogated by vasoactive intestinal peptide, which is a JNK inhibitor; however, colocalization of activated JNK (p-JNK) and activated caspase-3 did not clearly identify JNK as an upstream activator of caspase-3. We propose that these changes are indicative of cell death in aged hippocampus and suggest that they contribute to the age-related decrease in long-term potentiation in perforant path granule cell synapses. PMID- 12081658 TI - In vivo regulation of dopamine and noradrenaline release by alpha2A-adrenoceptors in the mouse prefrontal cortex. AB - The present study investigated the role of alpha2A-adrenoceptor subtype in the regulation of noradrenaline and dopamine release in the medial prefrontal cortex. The effect of local introduction of the alpha2-adrenoceptor agonist dexmedetomidine (10-9-10-8 m) on noradrenaline and dopamine release was investigated in alpha2A-adrenoceptor knockout and control mice by using in vivo microdialysis. Furthermore, to reveal a possible distinction between regulation of baseline and peak release, we sampled the dialysate during both rest and handling-induced mild stress. Baseline noradrenaline and dopamine concentrations did not differ between alpha2A-adrenoceptor knockout and control mice. Dexmedetomidine decreased, in a concentration-dependent manner, noradrenaline and dopamine levels in both genotypes. However, the effect of dexmedetomidine on noradrenaline release was attenuated in the alpha2A-adrenoceptor knockout mice, whereas the effect on dopamine release did not differ between the genotypes. The first handling episode increased noradrenaline and dopamine levels to the same extent in both genotypes. However, in alpha2A-adrenoceptor knockout mice the noradrenaline and dopamine levels remained elevated in the samples following the first handling whilst, in the control mice, transmitter levels returned to baseline levels. In control mice the handling-induced peak noradrenaline and dopamine levels were lower after the administration of dexmedetomidine than during the first handling episode, but in alpha2A-adrenoceptor knockout mice no drug effect on handling-induced peak noradrenaline and dopamine levels was found. Our results suggest that the release of noradrenaline in the medial prefrontal cortex is mainly regulated via alpha2A-adrenoceptors, whilst other alpha adrenoceptor subtypes play a significant role in the regulation of dopamine release. PMID- 12081659 TI - Cannabinoids inhibit excitatory inputs to neurons in the shell of the nucleus accumbens: an in vivo electrophysiological study. AB - The nucleus accumbens (NAc) represents a critical site for the rewarding properties of diverse classes of drugs of abuse. Glutamatergic afferents to the NAc are involved in the actions of psychostimulants and opioids, while the potentiation of dopaminergic neurotransmission in the NAc is a common feature of abused drugs, including cannabinoids. Cannabinoid receptors (CB1) are densely expressed in regions that provide excitatory innervation to the NAc, such as the amygdala, the cortex and the hippocampus. Recent in vitro evidence suggests that indeed cannabinoids modulate glutamatergic synapses in the NAc. In this study we recorded extracellularly from neurons in the shell of the NAc which responded to the stimulation of the baso-lateral amygdala (BLA) or the medial prefrontal cortex (PFC) in urethane anaesthetized rats. BLA or PFC stimulation induced generation of action potentials in NAc neurons. This excitatory effect was strongly inhibited by the synthetic cannabinoid agonists WIN 55212,2 (0.062-0.25 mg/kg, i.v.) and HU-210 (0.125-0.25 mg/kg, i.v.) or the psychoactive principle of Cannabis delta(9)-tetrahydrocannabinol (1.0 mg/kg, i.v.). Neither the D1 or D2 dopamine receptor antagonists (SCH23390 0.5-1.0 mg/kg, sulpiride 5-10 mg/kg, i.v.) or the opioid antagonist naloxone (1.0 mg/kg, i.v.) were able to reverse the action of cannabinoids, while the selective CB1 receptor antagonist/reverse agonist SR141716A (0.5 mg/kg, i.v.) fully suppressed the action of cannabinoid agonists, whereas per se had no significant effect. These results provide evidence that cannabinoids, in common with other drugs of abuse, in vivo strongly inhibit the excitability of neurons in the shell of the NAc. PMID- 12081660 TI - Effect of systemic nitroglycerin on CGRP and 5-HT afferents to rat caudal spinal trigeminal nucleus and its modulation by estrogen. AB - Systemic administration of nitroglycerin, a nitric oxide donor, triggers in migraine patients a delayed attack of unknown mechanism. After puberty migraine is more prevalent in women. Attacks can be triggered by abrupt falls in plasma estrogen levels, which accounts in part for sexual dimorphism, but lacks an established neurobiological explanation. We studied the effect of nitroglycerin on the innervated area of calcitonin gene-related peptide (CGRP) and serotonin immunoreactive afferents to the superficial laminae of the spinal portion of trigeminal nucleus caudalis, and its modulation by estrogen. In male rats, nitroglycerin produced after 4 h a significant decrease of the area innervated by CGRP-immunoreactive afferents and an increase of that covered by serotonin immunoreactive fibres. These effects were not observed in the superficial laminae of thoracic dorsal horns. The effect of nitroglycerin was similar in ovariectomized females. In estradiol-treated ovariectomized females the area in the spinal portion of trigeminal nucleus caudalis laminae I-II covered by CGRP immunoreactive fibres was lower and that of serotonin-immunoreactive fibres was higher than in males and for both transmitters not significantly changed after nitroglycerin. The bouton size of CGRP profiles was smaller in estradiol-treated ovariectomized females, whereas after nitroglycerin it decreased significantly but only in males and ovariectomized females. Nitroglycerin, i.e. nitric oxide, is thus able to differentially influence afferent fibres in the superficial laminae of rat spinal trigeminal nucleus caudalis. Estradiol modulates the basal expression of these transmitters and blocks the nitroglycerin effect. These data may contribute to understanding the mechanisms by which estrogens influence migraine severity and the triggering of attacks by nitric oxide. PMID- 12081662 TI - NMDA receptor-mediated metaplasticity during the induction of long-term depression by low-frequency stimulation. AB - Metaplasticity refers to the activity-dependent modification of the ability of synapses to undergo subsequent synaptic plasticity. Here, we have addressed the question of whether metaplasticity contributes to the induction of long-term depression (LTD) by low-frequency stimulation (LFS). The experiments were conducted using standard extracellular recording techniques in stratum radiatum of area CA1 in hippocampal slices made from adult Sprague-Dawley rats. The degree of LTD induction was found to be a nonlinear function of the number of pulses during a 1-Hz LFS. Little LTD was observed following 600 or 900 pulses, but a significant LTD occurred following 1200 pulses of LFS, whether delivered in one episode, or in two bouts of 600 pulses given 10 min apart. A similar pattern was observed for 3 Hz LFS. The data support the suggestion that pulses occurring early in the LFS train prime synapses for LTD induction, as triggered by later occurring stimuli. The priming effect lasted at least 120 min, when tested by giving two bouts of 1 Hz LFS (600 pulses each) at different intervals. Neither heterosynaptic nor homosynaptic stimulation by itself was sufficient to prime LTD. However, a combination of the stimuli, induced by increased stimulus strength during the LFS, appeared necessary for inducing the effect. An N-methyl d-aspartate (NMDA) receptor antagonist markedly reduced total LTD induction, regardless of whether it was administered during the first or second LFS in a protocol employing two bouts of 600 pulse LFS, 30 min apart. These findings strongly support the hypothesis that NMDA receptor-dependent metaplasticity processes contribute to the induction of LTD during standard LFS protocols. PMID- 12081661 TI - Acute and long-term changes in the mesolimbic dopamine pathway after systemic or local single nicotine injections. AB - We have examined several neurochemical and behavioural parameters related to the function of the mesolimbic dopamine (DA) pathway in animals treated with nicotine following three modes of drug administration, i.e. systemic intraperitoneal injection, intra-accumbens (Acb) infusion or intraventral tegmental area (intra VTA) microinjection. The present modes of systemic, intra-Acb and intra-VTA nicotine administration elicited comparable acute increases in dialysate DA levels from the Acb. The increase in extracellular DA levels was paralleled by a significant enhancement of locomotion in a habituated environment in the case of systemic or intra-VTA nicotine administration, whereas unilateral or bilateral intra-Acb nicotine infusion was ineffective, showing that accumbal DA increase is not sufficient to elicit locomotion in this experimental paradigm. Intra-VTA, but not systemic or intra-Acb, nicotine administration caused a long-term (at least 24-h) increase in basal dialysate DA levels from the Acb. In addition, significant increases in tyrosine hydroxylase (TH) and GluR1 (but not dopamine transporter or NR1) mRNA levels in the VTA were detected 24 h after intra-VTA nicotine administration. Systemic nicotine injection caused only an increase in TH mRNA levels while intra-Acb infusion did not modify any of the mRNAs tested. The long-term increase in basal DA levels in the Acb and TH, and GluR1 mRNA levels in the VTA upon intra-VTA nicotine microinjection indicates that even a single nicotine injection can induce plastic changes of the mesolimbic DA pathway. PMID- 12081663 TI - Donepezil reverses a mnemonic deficit produced by scopolamine but not by perforant path lesion or transient cerebral ischaemia. AB - The purpose of these studies were threefold. Firstly, to further characterize the effect of perforant path transection on a test of short-term memory: delayed matching (or nonmatching)-to-position [D(N)MTP]. Secondly, to evaluate the effect of a transient cerebral ischaemia in the same task. Both surgical procedures were chosen as they produce a CNS lesion similar to that described in Alzheimer's Disease (AD). Thirdly, the effect of the acetylcholinesterase inhibitor, donepezil (Aricept(R), E2020), on the resulting cognitive impairment was studied. Perforant path transection produced a robust, delay-dependent impairment of choice accuracy in rats performing either a delayed matching- or nonmatching-to position task. Sample latency was also reduced following lesion, yet the lesion induced impairment was not affected by increasing the response requirement at the sample stage. An 11-min period of transient ischaemia (two-vessel occlusion model) resulted in almost complete loss of hippocampal CA1 pyramidal cells and a delay-dependent impairment in DMTP performance. However, unlike perforant path lesions, this deficit was unstable and declined in magnitude over the experimental period. Increasing the delay interval restored this deficit. Donepezil, at doses that robustly attenuated a scopolamine (0.06 mg/kg s.c.) induced DMTP accuracy impairment in naive, unoperated rats, had no effect against either lesion-induced impairment. The results are considered in terms of the effectiveness of acetylcholinesterase inhibitors in noncholinergic-based preclinical cognitive models. PMID- 12081664 TI - The basolateral complex of the amygdala is necessary for acquisition but not expression of CS motivational value in appetitive Pavlovian second-order conditioning. AB - The basolateral complex of the amygdala (ABL) is involved in processing information about stimulus motivational value. However, it is not clear whether the ABL is critical for acquisition, maintenance, or expression of this information. Our previous work has shown that ABL lesions made prior to training, block acquisition of an appetitive Pavlovian second-order conditioning task, in which performance is thought to depend on the acquisition of motivational (conditioned reinforcement) value by the first-order conditioned stimulus (CS). The present experiments examined the effects of ABL lesions made after first order conditioning, when the CS acquires motivational value, but before second order conditioning, the test for acquired value of that CS. Rats received pairings of a visual CS with a food reinforcer. They then received bilateral sham or excitotoxic lesions of the ABL. After recovery, they received pairings of a second-order auditory CS with the previously conditioned visual CS. In two experiments, both sham and lesioned rats acquired normal second-order conditioned behaviours. Some of the same rats were then given another round of second-order conditioning with novel CSs. In this case, when first-order training occurred after surgery, some second-order conditioned behaviours were impaired in lesioned rats. Tests of the associative underpinnings of second-order conditioned behaviours showed that those behaviours impaired by ABL lesions were based on stimulus-response associations. The results show that although the ABL is critical for second-order conditioning, this role is limited to acquisition of information about the motivational value of the first-order CS, and it is not critical for maintenance of this information or for its use in forming second order associations. PMID- 12081665 TI - Multiple zonal projections of the nucleus reticularis tegmenti pontis to the cerebellar cortex of the rat. AB - Compartmentalization (alternating labelled and unlabelled stripes) of mossy fibre terminals was found in the cerebellar cortex after iontophoretic injections of biotinylated dextran amine into discrete regions of the nucleus reticularis tegmenti pontis (NRTP). The zonal pattern was only observed when volumes of nuclear tissue ranging from 4.5 x 106 to 17.66 x 106 microm3 were impregnated. Up to nine compartments (i.e. up to five stripes separated by four interstripes) were found in crus I and in vermal lobule VI. Up to seven compartments (four stripes and three interstripes) were found in crus II; up to five compartments (three stripes and two interstripes) were identified in the lobulus simplex, the paraflocculus and vermal lobules IV, V and VII; up to three compartments (two stripes and one interstripe) were identified in the paramedian lobule and, finally, up to two compartments (one stripe and one interstripe) were identified in the copula pyramidis, in the flocculus and in vermal lobules II, III, VIII and IX. The projections of the NRTP are arranged according to a divergent/convergent projection pattern. From single injections in the NRTP, projections were traced to a set of cortical stripes widely distributed over the cerebellar cortex. The set of stripes labelled from different regions of the NRTP partially overlapped but complete overlap was never found. This finding revealed that the topographic combination of the projections of the NRTP to the cerebellar cortex is specific for each region of the NRTP. Finally, the projections to single cortical areas were arranged according to a pattern of compartmentalization that is specific for each cortical area, independent of the site of injection in the NRTP and of the number of stripes evident in the cortex. PMID- 12081666 TI - Rapid regrowth of hippocampal mossy fibres and preceding maturation of NMDA receptor-mediated neurotransmission. AB - Early in postnatal development, glutamatergic synapses contain primarily NMDA receptors and progressively acquire AMPA receptor function. To determine whether this transformation occurs in a process of regenerative synaptogenesis following axotomy, we investigated the recovery of AMPA and NMDA receptor-mediated neurotransmission after the transection of mossy fibres (MF) in organotypic hippocampal cultures. An NMDA component could already be elicited 1 day after the lesion and reached a saturated level after 3 days. Thereafter, an AMPA component appeared and slowly matured after 10 days. The preceding establishment of NMDA receptor function implies that immature MF synapses are functionally silent at least for the first several days of recovery. The appearance of AMPA receptor mediated neurotransmission was unchanged in the presence of an NMDA-receptor antagonist or tetrodotoxin, which suggests that the AMPA receptor maturation is virtually independent of neuronal activity. Thus, the conversion of silent to functional synapses is not unique to synaptic plasticity or developmental processes but also occurs in recovery after brain damage, but its mechanism is likely to differ from NMDA receptor-dependent recruitment of AMPA receptors in synaptic plasticity. PMID- 12081667 TI - Selective enhancement of spatial learning under chronic psychosocial stress. AB - The hippocampus has long been proved to be implicated in several learning and memory processes. Being integrated into the limbic-hypothalamus-pituitary-adrenal axis, the hippocampus also plays an active role in the regulation of the stress response. Long lasting elevated levels of glucocorticoids resulting from a prolonged stress exposure affect hippocampal functions and structure, inducing learning and memory alterations and suppressing cell proliferation in the adult dentate gyrus. Here, adult male tree shrews (Tupaia belangeri) exposed to chronic psychosocial stress were tested repeatedly on a holeboard apparatus using two different learning tasks devised to evaluate hippocampal-dependent and hippocampal-independent cognitive function. We show that chronic stress enhanced learning in animals performing the hippocampal-dependent task, whereas no stress induced effect was found in the hippocampal-independent task. Additionally, after five weeks of stress, cell proliferation was reduced in the hippocampal dentate gyrus. These results indicate that specific memory processes not only may remain intact, but indeed are facilitated by chronic stress, despite elevated cortisol levels and suppressed hippocampal cell proliferation. PMID- 12081668 TI - Use of alternative therapies by patients undergoing surgery for nonmelanoma skin cancer. AB - BACKGROUND: The use of alternative therapies is becoming increasingly common and may result in unwanted side effects and drug interactions. OBJECTIVE: To determine the frequency of alternative medicine use by patients undergoing Mohs surgery for nonmelanoma skin cancer. METHODS: A written survey concerning use of alternative therapies was given to patients undergoing Mohs surgery for nonmelanoma skin cancer. A follow-up telephone survey was used to clarify positive responses. RESULTS: Thirty-six of 192 (18.8%) patients with nonmelanoma skin cancer reported using some type of alternative therapy. Almost all patients were using these therapies for conditions unrelated to their nonmelanoma skin cancer. Herbs and botanicals were the most common type of alternative therapy utilized. CONCLUSION: Physicians performing Mohs surgery should be aware that a significant number of their patients use alternative therapies. Since most patients do not readily volunteer information about alternative medicine usage, specific steps should be taken to obtain this information. PMID- 12081669 TI - Dietary supplements in the setting of mohs surgery. AB - BACKGROUND: The use of dietary supplements has become increasingly popular. While many are safe in small doses, others may have potentially harmful effects, particularly in surgical patients. OBJECTIVE: To study the incidence of dietary supplement use in patients presenting for Mohs surgery. METHODS: One hundred consecutive patients presenting for Mohs surgery completed a questionnaire providing all current medications. During the consultation, the patients were then asked specifically about their current use of any dietary supplements. Responses differing from those on the questionnaire were recorded. RESULTS: Forty nine of 100 patients (49%) were currently taking dietary supplements. Of this group, 17 patients (35%) self-reported the use of supplements; 32 patients (65%) did not. Thirty women (59%) were currently using dietary supplements regularly compared to 19 men (39%). Women were also more likely to self-report the use of supplements compared to men: 14 women (47%) versus three men (15%). Forty-eight of the 100 study patients (48%) were currently taking anticoagulant medications such as aspirin, warfarin, nonsteroidal anti-inflammatory drugs (NSAIDs), or clopidogrel bisulfate. Fifty instances were noted where patients were taking one or more dietary supplements that have demonstrated anticoagulant properties. Of this group, 21 instances (42%) where patients took a combination of prescription and over-the-counter (OTC) anticoagulants and one or more dietary supplements shown to have effects on coagulation were recorded. CONCLUSION: Of the almost 50% of patients taking dietary supplements, one-third reported usage, while two thirds did not. Women used dietary supplements more frequently than men and were more than three times more likely to offer this information. Furthermore, many supplements have been shown to have effects on coagulation, including vitamin E, garlic, ginkgo, feverfew, and fish oils. Use of these substances alone or in combination may potentiate the anticoagulant effects of each other or prescribed medications. It is therefore important for the dermatologic surgeon to communicate openly with patients regarding dietary supplements to avoid potential complications during or following surgery. PMID- 12081670 TI - The laser-assisted neck lift: modifications in technique and postoperative care to improve results. AB - BACKGROUND: Conventional submental tumescent liposuction has proved disappointing for some patients with anterior neck laxity, ptotic platysma muscles, and increased subplatysmal fat. Many of these patients are facelift candidates but are unwilling to undergo this extensive procedure. We describe our hybrid approach, which offers consistently improved results and enhanced patient satisfaction. OBJECTIVE: To establish a sharper cervicomental angle by more completely removing subplatysmal fat. We also wished to achieve more consistent, smoother results, minimizing ripples, folds, and hematomas with a novel postoperative dressing system. METHODS: Extensive tumescent liposuction of the lower face, jowls, and anterior neck was performed. Following this, subplatysmal fat was removed by dissection, the platysma muscle was imbricated, and the CO2 laser utilized in a defocused, low-power mode to partially treat the dermal undersurface and underlying muscle. Our postoperative dressing included a 10 cm mineral oil polymer gel disc in the submental location, covered by tape, silicone foam, and a lower face and neck garment to provide both support and even compression over the entire neck for at least the first 24 hours. RESULTS: Results proved uniformly satisfying for most patients, even those in their senior years. Benefits included an improved cervicomental angle, a decrease in jowling, and a marked reduction in the laxity and wrinkling of the neck skin and horizontal neck creases. Problems related to postoperative rippling or folding of the redraped skin and hematoma formation were minimized. conclusion. Laser neck tightening combined with tumescent liposuction and an advanced postoperative dressing for superior support and uniform compression has resulted in consistently excellent outcomes with improved patient satisfaction. PMID- 12081671 TI - Lifestyle outcomes of tumescent liposuction surgery. AB - OBJECTIVE: To assess the lifestyle benefits of liposuction surgery using tumescent local anesthesia and to attempt to establish any significant factors affecting these lifestyle outcomes. METHODS: Standardized questionnaires were sent to 332 patients who had liposuction performed at the clinic in 1999 and 2000 for purely cosmetic indications. Questions were designed to evaluate specific lifestyle benefits generally attributed to liposuction surgery by intending patients and the general public. RESULTS: Of the 332 patients, 123 (37%) completed and returned the survey; 87.8% of respondents were female. The mean body mass index (BMI) was 26.16. A large proportion of patients experienced positive lifestyle outcomes from the procedure: 80.5% were more confident, 74.8% noted an increase in self-esteem, and 87% were more comfortable in clothes. The time since surgery did not influence results. CONCLUSION: Liposuction using the tumescent technique is associated with a wide range of beneficial lifestyle outcomes for the majority of patients. PMID- 12081672 TI - Microprocessor-controlled local anesthesia versus the conventional syringe technique in hair transplantation. AB - BACKGROUND: Local anesthesia by the traditional injection technique is central to optimizing patient comfort during hair transplant procedures. However, even when carefully and properly performed, some patients still describe their treatment sessions as painful. OBJECTIVE: To determine if patients undergoing hair transplantation experience less pain and discomfort when their local anesthesia is performed with the Wand (as of spring 2001, the Wand has been renamed "CompuMed- Featuring the Wand Handpiece"), a microprocessor pump that precisely controls the infusion rate and pressure of local anesthetic solution. METHODS: We used combined retrospective studies and a prospective blinded study of men and women who received hair transplantation treatments between January 1999 and March 2001 at our private outpatient clinics in the United States. A total of 101 patients compared their experiences during their hair transplant sessions performed by us before and after we started using the Wand. Thirty-nine patients compared their experiences during treatments done by 32 other medical groups and when we used the Wand. In addition, 88 patients compared the Wand to injection while blinded to technique. The outcomes were measured with patient pain rating questionnaires using the Wong-Baker Faces Pain Rating Scale (0, no pain-5, severe pain). RESULTS: Patients in all three study groups reported that anesthesia with the Wand was less painful. Wand anesthesia was overwhelmingly (68%) described as associated with none (0) or very little (1) pain. Surprisingly, Wand anesthesia was associated with superior comfort not only during the administration of anesthesia (P =.005), but throughout the treatment session and during the first 48 postoperative hours. CONCLUSION: Microprocessor-controlled local anesthesia with the Wand is superior to traditional injection in consistently producing comfortable hair transplant treatments. PMID- 12081673 TI - Reduction of conchal enlargement and/or anterolateral rotation: otoplasty by the cosmetic dermatologic surgeon. AB - BACKGROUND: Otoplasty is the correction of protuberant ears. One of the most common malformations responsible for prominauris is conchal enlargement and/or anterolateral rotation. The surgical technique we utilize for the correction of this type of auricular malformation is not unlike the removal of auricular skin and cartilage frequently performed by dermatologic surgeons in the treatment of cutaneous malignancies of the ear. OBJECTIVE: To introduce the reduction of conchal enlargement and/or anterolateral rotation for the correction of protuberant ears into the armamentarium of the cosmetic dermatologic surgeon. METHODS: We describe in detail the development and anatomy of the ear along with indications, preoperative considerations, surgical technique, postoperative care, and potential complications for this type of cosmetic surgery. RESULTS: The technique described herein is an effective approach in the surgical management of protuberant ears that are caused by conchal enlargement and/or anterolateral rotation. CONCLUSION: With proper patient selection, thorough knowledge of this step-by-step surgical approach to otoplasty, and some prior surgical experience involving the skin and cartilage of the external ear, the successful correction of protuberant ears is well within the scope of the cosmetic dermatologic surgeon's practice. PMID- 12081674 TI - Glycolic acid peels compared to microdermabrasion: a right-left controlled trial of efficacy and patient satisfaction. AB - BACKGROUND: Glycolic acid peels and microdermabrasion are widely recommended by dermatologists and others for facial skin rejuvenation, but few studies have assessed their clinical efficacy. OBJECTIVE: To compare the relative efficacy of glycolic acid peels and microdermabrasion for facial skin rejuvenation. METHODS: An unblinded, randomized controlled trial was used. Each of 10 patients received paired treatments with glycolic acid peels (20%) and microdermabrasion (mild setting) for 6 consecutive weeks. The right and left sides of the face were treated with different modalities. Once a patient was assigned to receive a particular modality to a particular side of the face, all subsequent treatments were delivered in the same manner. Patient ratings, investigator ratings, and photographs were obtained before the first treatment and 1 week after the last treatment. RESULTS: In terms of overall preference for a given type of treatment, seven patients chose glycolic acid peels, one selected microdermabrasion, and two had no favorite treatment. The relative preference for glycolic acid peels approached significance (P =.0578). Investigator ratings and photographic comparisons did not reveal treatment-specific differences or significant improvement from baseline. No serious side effects were seen with either treatment, and even mild effects were seldom reported. CONCLUSION: In this study, patients appeared to prefer low-strength glycolic acid peels to low-intensity microdermabrasion for facial rejuvenation. Differences in patient satisfaction were subtle and may be technique dependent. PMID- 12081675 TI - Long-term quantitative benefits of botulinum toxin type A in the treatment of axillary hyperhidrosis. AB - BACKGROUND: Although axillary hyperhidrosis is readily treated with botulinum toxin, the time course of benefits is not well established. OBJECTIVE: To quantify the long-term effectiveness of botulinum toxin type A (BTX-A) for the treatment of axillary hyperhidrosis. METHODS: This was a double-blind, placebo controlled study. Eighteen patients received intradermal injections of either 100 U BTX-A (50 U/ml/axilla) or placebo. Sweating per surface area was quantified monthly for 5 months. RESULTS: The BTX-A group had an average reduction in sweat production of 91.6% at 2 weeks (from 5.03 ml/min/m(2) to 0.42 ml/min/m(2), P <.05). The average reduction over 5 months was 88.2%. At the end of the study, only 1 of 12 BTX-A-treated patients had returned to baseline sweat production. CONCLUSION: These quantitative results demonstrate that BTX-A is a safe and effective treatment for axillary hyperhidrosis and that the benefits last for at least 5 months. PMID- 12081676 TI - Phagocytosis of different particulate dermal filler substances by human macrophages and skin cells. AB - BACKGROUND: Foreign substances have been introduced into the human body with varying degrees of success. Polymethylmethacrylate (PMMA) microspheres of different sizes recently have been manufactured for use as a filler substances in the skin and other organs. OBJECTIVE: To establish whether the size of PMMA microspheres determines whether various cell types initiate phagocytosis. METHODS: The capacity of three different cell lines-U-937 cells, XS 106 and XS 52 Langerhans cells, and HaCaT keratinocytes-to phagocytose microspheres of varying sizes was examined using light and confocal microscopy as well as fluorescence activated cell sorter (FACS) analysis. Tumor necrosis factor (TNF)-alpha secretion was also determined. RESULTS: The U-937 cells, keratinocytes, and Langerhans cells could phagocytose PMMA particles of 20 microm or smaller. Microspheres larger than 20 microm were not ingested by any of the cells. CONCLUSION: Microspheres larger than 20 microm have a lower likelihood of being phagocytosed. Thus this study suggests that microspheres 40-50 microm in diameter are less likely to initiate an inflammatory reaction when injected into the dermis and subdermis as a filler substance. On the other hand, microparticles made of silicone and polymethacrylate were phagocytosed, possibly because of their different structure. PMID- 12081677 TI - Safety data of injectable nonanimal stabilized hyaluronic acid gel for soft tissue augmentation. AB - BACKGROUND: Nonanimal hyaluronic acid gel was recently developed for soft tissue augmentation and volume expansion and has been shown to offer several advantages in comparison to other augmentation materials. There are rare reports of adverse events believed to be secondary to trace amounts of proteins in the hyaluronic acid raw material. OBJECTIVE: To determine the safety profile of nonanimal stabilized hyaluronic acid gel (Restylane, Perlane, Restylane Fine Lines, Q-Med AB, Uppsala, Sweden) for soft tissue augmentation using a retrospective review of all adverse events data from Europe, Canada, Australia, South American, and Asia from 1999 and 2000. RESULTS: Data from an estimated 144,000 patients treated in 1999 indicated the major reaction to injectable hyaluronic acid was localized hypersensitivity reactions, occurring in approximately 1 of every 1400 patients treated. In 1999 there was an adverse event reported for 1 of every 650 patients (0.15%) treated. These were temporary events that included redness, swelling, localized granulomatous reactions, bacterial infection, as well as acneiform and cystic lesions. For 2000 there was an estimated 262,000 patients treated with hyaluronic acid gel. The total number of adverse events was 144, corresponding to one adverse event for every 1800 patients (0.06%) treated. The major adverse event was again hypersensitivity, occurring in 1 of every 5000 patients treated. CONCLUSION: According to the reported worldwide adverse events data, hypersensitivity to nonanimal hyaluronic acid gel is the major adverse event and is most likely secondary to impurities of bacterial fermentation. According to data from 2000, the incidence of hypersensitivity appears to be declining after the introduction of a more purified hyaluronic acid raw material. PMID- 12081678 TI - Long-term efficacy and quality of life in the treatment of focal hyperhidrosis with botulinum toxin A. AB - BACKGROUND: Botulinum toxin A has been used increasingly in the treatment of focal hyperhidrosis. OBJECTIVE: To assess the long-term efficacy of botulinum toxin A in the treatment of hyperhidrosis and the changes in quality of life and patient satisfaction with treatment. METHODS: A questionnaire was designed to assess the efficacy using visual analog scales and the quality of life both before and after treatment using a modified Dermatology Life Quality Index scale. RESULTS: There was a reduction in the hyperhidrosis and a statistically significant improvement in the quality of life scores for the axillae, palms, and forehead. CONCLUSION: Botulinum toxin A injections are safe and effective for the treatment of hyperhidrosis of the axillae, palms, and forehead, resulting in an improved quality of life for patients. PMID- 12081679 TI - The distribution of follicular units in the Chinese scalp: implications for reconstruction of natural-appearing hairlines in Orientals. AB - BACKGROUND: Follicular transplantation using hair in its naturally occurring groups, called follicular units (FUs), has become the most popular technique in hair restoration surgery. Recently follicular transplantation was performed with a qualitative and quantitative concept to achieve the best clinical result. The characteristics and distribution of FUs are well studied in Caucasians and widely applied in hair transplantation surgery. OBJECTIVE: In order to understand the normal distribution of FUs in the Chinese scalp, we counted the number of hairs and FUs in normal Chinese scalp to provide general information for surgical planning and design in bald Chinese patients. METHODS: A total of 50 normal and 50 bald Chinese adults were enrolled to count the hairs on their scalp. One hundred bald patients receiving hairline reconstruction were also prospectively quantitatively evaluated. RESULTS: In normal Chinese scalp, an average 71.78 FUs/cm(2) and 137.08 hairs/cm(2) were calculated with a follicular density of 1.91 hairs/FU. Two-hair FUs are the predominate group (50.29%). In bald patients, an average of 68.07 FUs/cm(2) was found, which was less than that of the occipital scalp in normal nonbald patients. In reconstruction of the frontal hairline, a total of 700-1000 FUs were implanted with an average density of 30 FUs/cm(2). CONCLUSION: We found the average number of FUs (0.72 FU/mm(2)) was less than that in Caucasian patients (1 FU/mm(2)). The average density of 30 FUs/cm(2) implanted was suitable to reconstruct a natural frontal hairline in bald Chinese patients, which can achieve about 40% of normal hair density. Our results could provide the hair surgeon with general information about hair distribution on the Chinese scalp for surgical planning and design in their patients. PMID- 12081680 TI - Clinical improvement of solar lentigines and ephelides with an intense pulsed light source. AB - BACKGROUND: A noncoherent, broadband, intense pulsed light (IPL) source has been used for the symptoms of photoaging skin as a nonablative method. OBJECTIVE: To investigate the efficacy and tolerability of IPL in solar lentigines and ephelides on the face. METHODS: An open study was performed in patients with solar lentigines and ephelides who received three to five treatments of IPL. RESULTS: Forty-eight percent of patients had more than 50% improvement and 20% had more than 75% improvement. In the group of solar lentigines, 40% of patients showed more than 50% improvement and 16% had more than 75% improvement. Patients with small plaques of solar lentigines responded well, whereas patients with small + large and large plaques showed poor response. Patients with solar lentigines + ephelides and ephelides responded remarkably with 75% and 71% of patients, respectively, having more than 50% improvement. CONCLUSION: IPL was well tolerated and may be a new modality for the therapy of solar lentigines and ephelides. PMID- 12081681 TI - Recurrent ingrown big toenails are efficiently treated by CO2 laser. AB - BACKGROUND: Surgery for onychocryptosis has a high rate of recurrence. OBJECTIVE: To evaluate CO2 laser partial matricectomy for recurrent onychocryptosis. METHODS: One hundred ninety-six consecutive patients (predominantly teenagers) previously unsuccessfully treated by surgery underwent CO2 laser for recurrent onychocryptosis. After a digital nerve block and a simple partial nail plate avulsion, the laser was used (5 W, defocused 2 mm beam in continuous mode) to vaporize the matrix, the lateral horn, and the lateral nail groove, including local granulation tissue if present. Follow-up was at least 12 months. RESULTS: Three hundred forty-four matricectomies were performed. Disease was mostly at stage II and III, with severe local infection in 24 cases (12.2%). All wounds healed in 21.9 +/- 3.2 days, with no postoperative local infection or prolonged exudative drainage. Onychocryptosis reoccurred in 5 of 344 treated margins (1.45%, average 15 months), all after primary bilateral matricectomy. Spicules in the lateral nail groove occurred in 14 of 344 treated margins (4%, average 5.9 months), mostly after primary bilateral matricectomy (7 cases) and in infected margins (8 cases). One patient developed a neuroma in the lateral nail groove. CONCLUSION: CO2 laser is effective for the treatment of recurrent onychocryptosis. Bilateral matricectomy and local infection seem to be the predisposing factors for recurrence and postoperative spicule growth. PMID- 12081682 TI - Perioperative management of anticoagulants and platelet inhibitors for cutaneous surgery: a survey of current practice. AB - BACKGROUND: Perioperative management of therapy with anticoagulants or platelet inhibitors for patients having cutaneous surgery presents dilemmas for dermatologic surgeons. OBJECTIVE: To outline the current spectrum of practice for perioperative management. METHODS: Questionnaires were mailed to 504 dermatologic surgeons. Data included use of warfarin, aspirin, and nonsteroidal anti inflammatory drugs (NSAIDs) perioperatively and involvement of other physicians in making management decisions. RESULTS: Of the responding physicians, 83% routinely ask primary physicians or cardiologists for recommendations about perioperative management, 80% discontinue warfarin therapy perioperatively at least some of the time, 26% always discontinue aspirin therapy, 38% manage aspirin and NSAIDs in the same manner, and 53% withhold therapy with NSAIDs for less time than with aspirin. CONCLUSION: Dermatologic surgeons use various perioperative management strategies. Despite no published evidence of increased hemorrhagic risk with anticoagulant or platelet inhibitor therapy during cutaneous surgery, many physicians discontinue therapy perioperatively. PMID- 12081683 TI - Therapeutic response of a brother and sister with xeroderma pigmentosum to imiquimod 5% cream. AB - BACKGROUND: Xeroderma pigmentosum (XP) is an autosomal recessive disease marked by solar sensitivity, photophobia, early onset of freckling, and solar-induced cutaneous neoplastic changes. These patients can often develop hundreds of cutaneous tumors, making surgical therapy difficult. Imiquimod 5% cream has been shown to have activity in treating various cutaneous malignancies. OBJECTIVE: To examine the effectiveness and tolerability of imiquimod 5% cream in treating facial basal cell carcinomas (BCCs) in a brother and sister with XP. These patients were developing skin cancers faster than could be managed surgically and had failed 6 months of chemoprophylaxis with isotretinoin. METHODS: Imiquimod 5% cream was applied to the faces of these two patients as frequently as tolerated, with the goal of gaining control over the many clinically evident BCCs present on the faces of these siblings. We also examined whether we could reduce the rate of new neoplasm development. RESULTS: The brother in our study tolerated imiquimod 5% cream twice a day every day with minimal inflammatory response. He had clinical resolution of many of the BCCs present within the treatment area as well as shrinking of many of the remaining lesions. He has continued to produce new tumors at a substantially reduced rate relative to his pretreatment baseline. The sister in our study exhibited a severe inflammatory response to imiquimod 5% cream, with facial swelling and erosion of the treated area with application as infrequent as three times a week. In spite of the vastly different inflammatory response, her cutaneous tumors responded favorably to therapy as well. CONCLUSION: Imiquimod 5% cream was effective in treating facial BCCs in these siblings with XP. As well, we have noted a significant reduction in the development of new tumors within the imiquimod-treated area. The inflammatory response to this medicine was at opposite extremes among these two siblings. However, this did not appear to alter the therapeutic benefit of this therapy. PMID- 12081684 TI - Management of large surgical defects of the forehead and scalp by imbrication of deep tissues. AB - BACKGROUND: When facing surgical defects, the dermatologic surgeon follows certain basic principles that help reduce the inherent tension to allow for a better cosmetic outcome. These commonly include the use of undermining, releasing technique such as galeotomy if applicable, selection of suture material of appropriate tensile strength, and closure along relaxed skin tension lines. OBJECTIVE: To review the imbrication of deep tissues, another surgical principle aimed at wound tension reduction and widely utilized by cosmetic surgeons in forehead lifts and scalp reductions, as it applies to dermatologic surgeons in the repair of large surgical defects of the upper face and scalp. The latter may be utilized both with primary closure and with local flaps. METHODS: We describe in detail the technique of imbrication of deep tissues and provide illustrations for a better understanding of how to correctly use this surgical principle. RESULTS: The dermatologic surgeon has an additional tool, termed imbrication of deep tissues, available to aid in the closure of sizable wounds of the forehead and scalp as seen following Mohs surgery for cutaneous malignancies by providing deep tissue support. CONCLUSION: Imbrication of deep tissue is an effective tool that may be used by dermatologic surgeons in conjunction with the more commonly utilized basic surgical principles to enhance the cosmetic outcome in the closure of large, high-tension defects of the forehead and scalp. PMID- 12081685 TI - Granuloma faciale successfully treated with long-pulsed tunable dye laser. AB - BACKGROUND: Granuloma faciale has been treated in the past with different modalities but the majority have had the risk of scarring. OBJECTIVE: Based on the principle of selective photothermolysis, we considered using the newer long pulsed tunable dye laser for the treatment of granuloma faciale to target the vessels and minimize scarring. METHODS: Confirmation of the diagnosis by a punch biopsy of the lesion was followed by three treatments on separate occasions 6 weeks apart with the long-pulsed tunable dye laser. RESULTS: There was significant flattening of the lesions after two treatments, with complete clearing after the third. No scarring was detectable and there was no recurrence in the 9-month follow-up. CONCLUSION: We conclude that granuloma faciale may be successfully treated with the long-pulsed tunable dye laser with minimal risk of scarring, especially in cosmetically sensitive areas. PMID- 12081686 TI - Treatment of inverse psoriasis with the 308 nm excimer laser. AB - BACKGROUND: The treatment of inverse psoriasis can be both challenging and frustrating. Conventional topical and systemic treatments can be limited in terms of efficacy, acceptability and safety. The 308 nm excimer laser has been shown to be effective in the treatment of psoriatic plaques. METHODS: A patient with chronic inverse psoriasis was treated with 308 nm excimer laser using a 3.5 cm spot, a dosage of 2 minimal erythema doses and a pulse width of 30 ns. RESULTS: Complete clearance of the psoriatic lesions was obtained after 3 weeks of treatment. Remission duration was at least 6 months. CONCLUSION: The 308 nm excimer laser is safe and effective for the treatment of inverse psoriasis. Treatments are well tolerated with minimal side effects. PMID- 12081687 TI - Diagnosis, workup, and prognosis of cutaneous metastases of unknown primary origin. AB - BACKGROUND: Metastatic cancer of unknown primary (MCUP) accounts for 5-10% of presenting oncology patients. Premorbid identification of the primary source occurs in only 20-25% of cases. A small fraction of these tumors present in the skin and are thus likely to be encountered by dermatologists and dermatologic surgeons. OBJECTIVE: To present a practical schema for the diagnosis and evaluation of patients with MCUP. METHODS: Case report and literature review. CONCLUSION: The diagnosis of MCUP is associated with a poor prognosis, with median survival rates of less than 1 year. Efforts to identify subgroups of patients with understanding of these subgroups will enable early identification of patients with treatable disease and more favorable prognoses. PMID- 12081688 TI - Microcystic adnexal carcinoma (MAC) of the scalp with extensive pilar differentiation. AB - BACKGROUND: Microcystic adnexal carcinoma (MAC), or sclerosing sweat duct tumor, is an uncommon, locally aggressive tumor. It typically involves the upper lip or face of middle-aged adults and rarely involves the scalp. It is characterized histologically by both pilar and eccrine differentiation and is associated with frequent perineural invasion. MAC is frequently misdiagnosed because of its bland and asymptomatic clinical presentation. In addition, its defining histologic features may be missed with a superficial biopsy. OBJECTIVE: To describe a case of MAC in the scalp treated with Mohs surgery. We describe a case with extensive pilar differentiation and clear cell changes. In addition, the clinical and histologic characteristics of this rare neoplasm are reviewed. METHODS: Mohs micrographic surgery (MMS) was performed on this neoplasm. RESULTS: The tumor was successfully excised in one surgery (five stages) and is without evidence of recurrence at 18 months. CONCLUSION: We present a case of MAC of the scalp in a 64-year-old white man treated successfully with MMS. PMID- 12081689 TI - Lessons on dermoscopy. PMID- 12081690 TI - Attaching instruments on surgical drapes. PMID- 12081691 TI - A new tie-over method improves cosmetic results. PMID- 12081692 TI - Botulinum toxin type A as an effective adjuvant therapy for Hailey-Hailey disease. PMID- 12081693 TI - Mucous extravasation phenomenon following vermilionectomy. PMID- 12081694 TI - Hematomas are underrepresented in studies on complications of ambulatory phlebectomy. PMID- 12081695 TI - Risk assessment failures of chemicals commonly used in consumer products. PMID- 12081696 TI - An assessment of the comparative sensitization potential of some common isothiazolinones. AB - The isothiazolinones are known contact sensitizers. Data are presented which demonstrate that, in comparison with the chlorinated and dichlorinated compounds which share immunological cross-reactivity, the unchlorinated isothiazolinones have a lower potential for sensitization and no documented immunological cross reaction with the chlorinated isothiazolinones. PMID- 12081697 TI - Surveillance scheme for occupational skin disease in the Saarland, FRG. First report from BKH-S. AB - Occupational skin disease (OSD) is a significant public health concern. To keep OSDs under surveillance, a register of OSDs (BKH-S) was implemented in 1999 in the Saarland, FRG. We conducted a population-based register study to analyse all initial reports of OSDs reported to the BKH-S. 336 cases were prospectively assessed from July 1999 to June 2001. In 78% (n = 263) of the cases an occupational cause for the skin disease was stated, 91% (n = 240) of which fell within 16 occupational groups. This is equivalent to 6.8 cases per 10,000 workers per year within these occupational groups. Notably, young age groups (median age 35 years) were affected with a female predominance. The most frequent final diagnosis was irritant contact dermatitis (75%) followed by allergic contact dermatitis (34%) and atopic dermatitis (19%). The overall prevalence of occupationally relevant delayed-type reactors was 21% and of immediate-type reactors 17%. Work-related delayed-type sensitizations to potassium dichromate (n = 8) and immediate-type sensitizations to natural rubber latex (n = 11) were the most frequently observed. Questionnaire results showed frequent occupational skin contact with wet work (59%), detergents (48%), and disinfectants (38%). The second valid surveillance scheme for OSD in the FRG has been introduced and can be made available for further analysis. PMID- 12081699 TI - A skin sensitization safety assessment of a new bleach activator technology in detergent applications. AB - A new chemical called nonanoyl amido caproylacid oxybenzenesulphonate (NACAOBS) is being developed for use as a bleach activator in laundry detergents. Bleach activators, like NACAOBS, are typically used at levels between 2% and 6% in laundry detergents. NACAOBS is stable in aqueous solutions, but undergoes rapid perhydrolysis when combined with water and peroxygen bleach in laundry detergents. Animal testing demonstrated that NACAOBS, as a raw material, is a weak skin sensitizer. Clinical testing, including extended simulated laundry pretreatment, human repeat insult patch testing and home use testing was then undertaken, following sufficient reassurance of 1) the weak sensitization potential of the substance, 2) its rapid degradation in laundry wash solutions and, consequently, 3) low-to-negligible consumer dermal exposures to the native substance. Results confirmed the skin sensitization safety profile of laundry detergents containing NACAOBS, namely the absence of any reaction suggestive of contact sensitization (even under exaggerated dermal exposure conditions in a detergent matrix), and a skin compatibility profile comparable to that of current detergents. Further confirmation of the skin safety profile was obtained from a successful 12-month market test of a granular detergent containing 3.6% of the new substance, during which not a single adverse skin reaction was reported. In addition, NOBS (an oxybenzenesulphonate structural analogue to NACAOBS) has similar toxicological properties and has been safely marketed in detergents at similar levels for many years. It can be concluded that the likelihood of NACAOBS to induce skin sensitization or even elicit allergic reactions in consumer detergent use scenarios is negligible. PMID- 12081698 TI - Monitoring levels of preservative sensitivity in Europe. A 10-year overview (1991 2000). AB - A 10-year multicentre analysis of the frequency of sensitivity to common preservatives collected in 16 centres in 11 countries has shown stable but persisting high levels of sensitivity to formaldehyde and 5-chloro-2-methyl-4 isothiazolin-3-one + 2-methyl-4-isothiazolin-3-one (MCI/MI). It has also revealed a significant increase in the level of reactivity to methyldibromoglutaronitrile (MDBGN) from 0.7% in 1991 to 3.5% in 2000. The current high level of sensitivity to MDBGN requires an urgent safety re-evaluation and risk assessment update along with consideration of immediate lowering of use concentrations, especially in leave-on products. PMID- 12081700 TI - Evaluation of biophysical and clinical parameters of skin barrier function among hospital workers. AB - The goal of this study was to evaluate biophysically and clinically the hand skin of nurses working in the operating room in comparison with the hand skin of a control group of female administrative employees. 19 nurses and 14 employees were included in this trial. Transepidermal water loss (TEWL) and stratum corneum (SC) hydration were evaluated on 4 different test sites - dominant and non-dominant pulse, and dominant and non-dominant hand dorsum. The clinical score Dermatology Life Quality Index (DLQI) and subjective evaluation of skin sensitivity were also recorded. In comparison with the control group, TEWL values among nurses were significantly higher on all test sites, while SC hydration was significantly lower on 3 of the 4 test sites. In addition, clinical score showed the presence of irritant contact dermatitis (ICD). DLQI reported a higher level of disability among the nurses group, while the subjective evaluation of skin sensitivity was identical in both groups. The results suggest the presence of skin barrier alterations and clinical signs of chronic ICD on the hands of nurses working in operating room units. PMID- 12081701 TI - The sensitizing potential of various biocides in the guinea pig maximization test. AB - Biocides are chemicals used as preservatives added to most cosmetics and toiletries and to industrial water-based products such as cutting fluids, paints and glues. There are numerous biocide formulations used in water-based metalworking fluids. All these products, characterized by a low molecular weight, can react with endogenous proteins and may induce allergic contact dermatitis. Thus, numerous reports on occupational allergic contact dermatitis have been published. The purpose of this experimental study was to compare the sensitizing potential of the various classes of biocides used in metalworking fluids by means of the maximization test of Magnusson & Kligman. The results show that, from the 9 samples tested, 7 biocides induced sensitization varying in grade from moderate to strong, while 2 benzisothiazoline-based samples did not induce allergic contact dermatitis. PMID- 12081702 TI - Type IV allergy in the food processing industry: sensitization profiles in bakers, cooks and butchers. AB - Data about the impact of allergic contact dermatitis in food processing occupations are scarce. The objective of this study was to identify the most frequent allergens and sensitization profiles in employees of the food processing industry. The results of patch testing performed in 873 bakers, cooks and butchers suspected of having occupational allergic contact dermatitis from 33 dermatological departments of the Information Network of Departments of Dermatology (IVDK) from 1992 to 1999 were analysed. A final diagnosis of allergic contact dermatitis was made in 213 patients (24.4%). Qualitative and quantitative differences in allergen profiles could be identified. Significantly higher sensitization rates were found in employees in the food processing industry, compared to the total test population, for nickel sulphate (22.4% versus 17.2%, p < 0.0005), thiuram mix (4.9% versus 2.6%, p < 0.0005), formaldehyde (3.5% versus 2.1%, p < 0.005) and Compositae mix (6.2%, p < 0.0005). Significantly lower sensitization rates were found for thimerosal (4.5% versus 6.9%, p < 0.05). For patch testing in food workers, we recommend the standard, the rubber and the Compositae series, as well as patients' own products according to the individual history. PMID- 12081703 TI - Allergic contact dermatitis from phthalic anhydride/trimellitic anhydride/glycols copolymer in nail varnish. PMID- 12081704 TI - Brown palm pseudochromhidrosis. PMID- 12081705 TI - Occupational allergic contact dermatitis from cured and used casting sand. PMID- 12081706 TI - A second florist with dermatitis from Trachelium caeruleum. PMID- 12081707 TI - Ammonium thiolactate and thiolactic acid: important hairdressers' allergens? PMID- 12081708 TI - Allergic contact dermatitis from prime yellow carnauba wax and coathylene in mascara. PMID- 12081709 TI - Eyelid dermatitis and conjunctivitis as sole manifestations of allergy to nickel in an orthodontic appliance. PMID- 12081710 TI - Pigmented contact dermatitis from topical minoxidil 5%. PMID- 12081712 TI - Occupational allergic contact dermatitis from the extreme pressure (EP) additive zinc, bis ((O,O'-di-2-ethylhexyl) dithiophosphate) in neat oils. PMID- 12081711 TI - Allergic contact dermatitis from urea-formaldehyde resin in medium-density fibreboard (MDF). PMID- 12081713 TI - Contact allergy to methyldibromo glutaronitrile in Euxyl K 400 in a cosmetic cream for protection against a permanent wave solution. PMID- 12081714 TI - Inherited hypertrichoses. AB - Hypertrichosis is a rare condition characterized by excessive growth of hair (terminal, vellus or lanugo) in areas of the body that are not predominantly androgen dependent, and it is independent of age, race or sex. It can be congenital, late-onset, generalized, localized, inherited or acquired. More than 50 different OMIM entries related to hypertrichosis exist, few of them with a localized gene locus or with a candidate gene. The review of generalized hypertrichoses from a historical point of view, including a review of their clinical and genetic features, shows heterogeneity with at least nine different entities. A short analysis of other forms of hypertrichosis is presented. PMID- 12081716 TI - Attention-deficit/hyperactivity disorder (ADHD): feasibility of linkage analysis in a genetic isolate using extended and multigenerational pedigrees. AB - Segregation analyses converge in explaining the predisposition to attention deficit/hyperactivity disorder (ADHD) as the consequence of a major gene and exclude purely environmental or cultural transmission. As a result of the ADHD phenotype restrictions, collection of extended families or design of linkage studies using families has been extremely difficult and thus currently linkage studies have been performed using only concordant or discordant sib-pairs rather than large families. On the other hand, intergenerational studies are represented by the transmission disequilibrium test (TDT) using trios. We collected pedigree data on ADHD from the Paisa community from Antioquia, Colombia, a genetic isolate. The goal of this study was to genetically map a putative gene predisposing to ADHD in a set of 27 multigenerational Paisa families. Here we present the results of a power simulation using SIMLINK to detect linkage of ADHD. ADHD was assumed to be a dichotomous trait with incomplete penetrance and a phenocopy rate of 3% in males and 0.2% in females. We simulated cosegregation of the trait and a marker locus in our pedigrees. We assumed Hardy-Weinberg and linkage equilibrium, equally frequent marker alleles and evaluated power at several recombination fractions between the trait and marker loci. Also, the ADHD trait was assumed to be genetically heterogeneous and different functions of age dependent penetrance were simulated. We found exceptionally good power to detect linkage (expected LOD > 14 if theta is 0.1 or less), and that the presence of heterogeneity up to 50% does not affect substantially the projected LOD scores even for a theta recombination value of 0.05 (eLOD > 5.87). Having now obtained blood samples and confirmatory interviews in five families (representing 20% of the projected number of families), we performed a new analysis. The expected mean LOD in these five families reached values close to 10 and remained invariant when heterogeneity and different penetrance models were considered. We discuss the relative benefits of using extended and multigenerational families for genetic mapping studies as opposed to using nuclear families, affected sib pairs or sporadic cases which require the collection of over 1000 analytical units to get the same power exhibited by the small number of pedigrees described here. PMID- 12081717 TI - Inducible nitric oxide synthase gene and diabetic retinopathy in Asian Indian patients. AB - Nitric oxide, a signal transduction molecule, when modulated causes various diseases including diabetic retinopathy. In diabetes, allelic polymorphism of the inducible nitric oxide synthase (iNOS) gene is associated with retinopathy in the Northern Irish population. In the present study we investigated the Asian Indian population. One hundred and ninety-nine unrelated Asian Indian patients with 15 or more years of type 2 diabetes were divided into two groups: (a) diabetic retinopathy (DR) and (b) diabetic nonretinopathy (DNR) subjects. In these groups the pentanucleotide microsatellite repeat located 2.5 kb upstream of the transcription start site of the iNOS gene was amplified by polymerase chain reaction and analyzed. Eleven alleles, 175-225 bp, were identified. Allele 210 bp was significantly associated with retinopathy (p = 0.044). Individuals carrying this allele had twice the risk of developing retinopathy compared with those who did not carry this allele [odds ratio (OR) - 2.03; 95% CI 0.96-4.35]. Alleles 200 and 220 bp were also significantly associated with no retinopathy and no serious retinopathy complications, respectively. In the Asian Indian population, allele 210 bp of the iNOS gene is a high-risk allele for developing retinopathy and alleles 200 and 220 bp protect an individual from developing retinopathy or its complications. PMID- 12081718 TI - New mutations in the CBFA1 gene in two Mexican patients with cleidocranial dysplasia. AB - Cleidocranial dysplasia (CCD) is an autosomal dominant skeletal disorder exhibiting a wide clinical spectrum ranging from minimal anomalies to classic CCD. Mutations scattered throughout the entire CBFA1 gene have been related to this disorder. However, it seems that most of them affect the highly conserved Runt domain, abolishing the DNA-binding ability of this transcription factor. Moreover, no systematic effect has been found to relate the type of mutation to the severity of the clinical features. In this paper, we studied two unrelated patients with classic CCD. DNA analysis revealed two novel mutations and three undescribed polymorphisms. One of the substitutions was a missense mutation in the Q/A domain leading to the replacement of a polar residue by a nonpolar one (158 A --> T [Q53L]). The second was an uncommon heterozygous stop codon mutation (1565 G --> C [X522S]) which theoretically results in a longer protein with 23 additional amino acids. This is the first report of this type of mutation in CBFA1. We discuss the possible consequences of these mutant sequences, although no phenotype-genotype correlation could be established. Our findings expand the existing number of allelic variants in this pathology. PMID- 12081719 TI - Deafness resulting from mutations in the GJB2 (connexin 26) gene in Brazilian patients. AB - Congenital deafness occurs in approximately 1 in 1000 live births. In developed countries about 60% of hearing loss is genetic. However, in Brazil most cases of hearing loss are due to environmental factors, such as congenital infections (mainly rubella), perinatal anoxia, kernicterus and meningitis. Recently, it has been demonstrated that the GJB2 gene is a major gene underlying congenital sensorial deafness. Mutations in this gene cause 10-20% of all genetic sensory hearing loss. One specific mutation, 35delG, accounts for the majority of mutant alleles. The extent of the hearing impairment varies from mild/moderate to profound, even within the patients homozygous for the common 35delG mutation. There may also be progression with age. Mutation analysis in the GJB2 gene was performed on 36 families (group A) presenting with at least one individual with non-syndromic deafness (NSD). An unselected series of 26 deaf individuals referred by other services where the environmental factors were not completely excluded was also part of the study (group B). Mutations in the GJB2 gene were found in 22% (eight patients) of the families tested in group A, and 11.5% (three patients) of individuals within group B. This finding should facilitate diagnosis of congenital deafness and allow early treatment of the affected subjects. PMID- 12081720 TI - De novo MECP2 frameshift mutation in a boy with moderate mental retardation, obesity and gynaecomastia. AB - Rett syndrome (RTT) is an X-linked neurodevelopmental disorder caused by mutations in the MECP2 gene, with apparent lethality in male embryos. However, recent studies indicate that mutations in the MECP2 gene can cause congenital encephalopathy, an Angelman-like phenotype and even nonspecific mental retardation in males. We report on a 10-year-old boy with moderate mental retardation, hypotonia, obesity and gynaecomastia and a de novo 2-bp deletion in the MECP2 gene that resulted in a frameshift and premature stop codon. As some of the clinical features were suggestive of the Prader-Willi syndrome, it might be worthwhile screening for MECP2 mutations in patients with an atypical Prader Willi phenotype but without the characteristic abnormalities on chromosome 15q. This report contributes to the phenotypic knowledge of male patients with MECP2 mutations. Moreover, this is the first reported male case of a de novo MECP2 mutation. PMID- 12081721 TI - Angiotensinogen genotype, plasma protein and mRNA concentration in isolated systolic hypertension. AB - Isolated systolic hypertension (ISH) occurs predominantly in the elderly, with a considerable morbidity and mortality. Its etiology is unknown but is likely to involve a significant genetic component. The aim of this study was to examine the angiotensinogen gene in ISH. The M235T and G(- 6)A polymorphisms were genotyped by polymerase chain reaction (PCR) in 86 ISH patients and 120 normotensive controls. Plasma angiotensinogen concentration was determined in 198 subjects by an indirect radioimmunoassay technique. Angiotensinogen mRNA concentration was determined by quantitative competitive reverse transcription (RT)-PCR in subcutaneous adipose tissue from a subset of these patients (n = 8) and controls (n = 6). Both the M235T (p = 0.0015) and G(- 6)A (p = 0.029) polymorphisms were associated with ISH. Plasma angiotensinogen concentration was higher in patients than controls (p < 0.0001), but was not associated with genotype. Angiotensinogen mRNA concentration in adipose tissue from ISH subjects was significantly lower than in adipose tissue from normotensive subjects (p = 0.033). The association of angiotensinogen gene variants with ISH and the elevation of plasma angiotensinogen concentration in these patients suggests a role of the angiotensinogen gene in this form of hypertension. Angiotensinogen gene expression may be altered in ISH, but this requires further examination. PMID- 12081722 TI - Familial lethal skeletal dysplasia with cloverleaf skull and multiple anomalies of brain, eye, face and heart: a new autosomal recessive multiple congenital anomalies syndrome. AB - We report on a 'new' lethal familial short-limb bone dysplasia associated with multiple anomalies in three sibs born to Arabic-Muslim consanguineous healthy parents. Clinical abnormalities included short limbs and short hands, cloverleaf skull, frontal bossing, wide anterior fontanel, hypertelorism, bilateral microphthalmia, cataract, low-set ears, narrow chest, ambiguous genitalia, cardiac ventricular septal defect (VSD) and agenesis of the corpus callosum. Radiological abnormalities included cloverleaf skull, hypoplastic clavicles and scapulae, thin, wavy cupped ribs, flat vertebral bodies with coronal clefting and several unossified vertebral pedicles and hypo-ossification of the pubic bone. The main changes noted in the long bones consisted of short-bowed long bones with abnormal metaphyses and unossified epiphyses. Chondro-osseous morphology documented degenerating chondrocytes with disorganization of the hypertrophied cartilage and short disorganized columns of hypertrophied areas. An autosomal recessive mode of inheritance seems most likely. PMID- 12081723 TI - A patient homozygous for the SCA6 gene with retinitis pigmentosa. AB - The present authors studied a 55-year-old-patient homozygous for the SCA6 gene who experienced frequent attacks of positional vertigo at 37 years of age with subsequent staggering gait and night blindness. Retinitis pigmentosa (RP), as well as cerebellar ataxia and vertical antidirectional nystagmus, were detected. The subject's parents were first cousins, and two of his three male cousins, whose parents were also first cousins, had RP without ataxia or nystagmus. The numbers of CAG repeats in the expanded alleles of the SCA6 gene found by molecular analysis were 21 and 21. The genetic results were negative for SCA1, SCA2, SCA3, SCA7 and dentatorubral pallidoluysian atrophy. The retinal degeneration in this patient is most likely to be secondary to a genetic disorder of autosomal or X-linked recessive inheritance rather than SCA6. Other reported cases of patients homozygous for the SCA6 gene are also reviewed. PMID- 12081724 TI - Recurrent 22q11.2 deletion in a sibship suggestive of parental germline mosaicism in velocardiofacial syndrome. AB - Deletions of chromosome 22q11.2 are recognized as the main cause of a number of clinical phenotypes, including velocardiofacial syndrome (VCFS) and DiGeorge syndrome (DGS). Velocardiofacial syndrome is a relatively common developmental disorder that is characterized by craniofacial anomalies and conotruncal heart defects. Most 22q11.2 deletions occur sporadically, although the deletion may be transmitted in some cases. The present performed a molecular analysis in one family including a patient with clinical diagnosis of VCFS and his sister with a suggestive phenotype. Six polymorphic 22q11.2 markers (i.e. D22S420, D22S264, D22S941, D22S306, D22S425 and D22S257) were used for genotype analysis of the DNA from the patients and unaffected relatives. The results revealed a 22q11.2 deletion in the patient and his sister from one of six markers (i.e. D22S941). Genotype analysis demonstrated that the deletion in this sib was of maternal origin. The results suggest that the mother probably has gonadal mosaicism. The other relatives present normal DNA profiles for all markers. These results have implications for genetic counseling because of a risk of transmission by germ cells carrying the deletion, even when parents present with a normal DNA profile in their blood cells. PMID- 12081725 TI - MECP2 mutations in Swedish Rett syndrome clusters. PMID- 12081726 TI - Familial pericentric inversion of chromosome 4: inv(4)(p16.1q12). PMID- 12081727 TI - High prevalence of I179S mutation in patients with late-onset metachromatic leukodystrophy. PMID- 12081728 TI - Early development of megarectum in myotonic dystrophy. PMID- 12081729 TI - Contrast-enhanced magnetic resonance angiography. PMID- 12081730 TI - Patient volumes and complications in thyroid surgery. PMID- 12081731 TI - Genetic pathways involved in the progression of Barrett's metaplasia to adenocarcinoma. AB - BACKGROUND: The prediction of which patients with Barrett's metaplasia will develop cancer is difficult. Better genetic characterization of the condition may aid clinicians in devising more effective management and follow-up strategies. METHODS: A review was undertaken of the accumulated genetic data relating to the progression of squamous epithelium to adenocarcinoma. The normal functions of a number of cancer-related genes are described and an explanation is given of how alterations in these genes interfere with normal cell processes and lead to cancer. RESULTS AND CONCLUSION: The main genetic alterations accompanying the progression through dysplasia to adenocarcinoma were collated from 135 papers. The principal genetic changes implicated are the loss of p16 gene expression (by deletion or hypermethylation), the loss of p53 expression (by mutation and deletion), the increase in cyclin D1 expression, the induction of aneuploidy and the losses of the Rb, DCC and APC chromosomal loci. PMID- 12081732 TI - Homocysteine and occlusive arterial disease. AB - BACKGROUND: An increased plasma level of homocysteine has been proposed as an independent risk factor for atherosclerosis; this review examines the evidence. METHODS: A Medline search was undertaken for English language articles on homocysteine and vascular disease. Further papers were identified by cross referencing from the reference lists of relevant major articles. RESULTS: Although much interest has been generated about homocysteine and atherosclerotic disease, contradictory data exist regarding its role in disease progression. There is insufficient current evidence to regard increased homocysteine level as a causative factor in atherosclerotic disease. CONCLUSION: It is not known whether lowering plasma homocysteine concentration will reduce cardiovascular risk in the long term. Until such data become available, there is no evidence for the widespread use of folic acid therapy to reduce cardiovascular disease risk. PMID- 12081733 TI - The colorectal adenoma-carcinoma sequence. AB - BACKGROUND: It is widely accepted that the adenoma-carcinoma sequence represents the process by which most, if not all, colorectal cancers arise. The evidence supporting this hypothesis has increased rapidly in recent years and the purpose of this article is to review this evidence critically and highlight its clinical significance. METHODS: Medline searches were used to identify recent key articles relating to the adenoma-carcinoma sequence. Further pertinent articles were obtained by manual scanning of the reference lists of identified papers. RESULTS: The evidence supporting the adenoma-carcinoma sequence can be classified as epidemiological, clinicopathological and genetic. The most recent and largest body of data relates to molecular genetic events and their cellular effects; however, many other approaches, such as cytogenetics, molecular cytogenetics and cytometry, have also yielded valuable information. CONCLUSION: Recent work continues to support the adenoma-carcinoma sequence, but there is a paucity of data on the interrelationship between different genetic mutations and on the relationship between molecular and other types of genetic abnormalities. The clinical utility of the observations described has yet to be fully realized and global genetic analysis of colorectal tumours may prove to be central in rational adenoma management. PMID- 12081734 TI - Late results concerning feasibility and compliance from a randomized trial of ultrasonographic screening for abdominal aortic aneurysm. AB - BACKGROUND: The study was an update at 10 years of a randomized trial of the efficacy of screening for abdominal aortic aneurysm (AAA). The extent of benefit, feasibility and compliance were examined, and reasons why this intervention may fail a proportion of those screened were identified. METHODS: A total of 6058 men aged 65 years and over were randomized to a group invited to attend ultrasonographic screening or to a control group. The mortality rate from AAA in the two arms of the trial was compared using a Poisson model. Analyses were by intention to treat. RESULTS: There was a 21 per cent reduction in mortality rate from AAA over the 10-year follow-up (relative risk 0.79 (95 per cent confidence interval 0.53 to 1.40)). The observed relative mortality reduction peaked at 4 years with a 52 per cent reduction in the study group. Eighteen of 24 AAA deaths in the study group were among those who did not attend the first screen, or failed to comply with the follow-up protocol. CONCLUSION: A greater awareness of the benefits of full participation in a screening programme could provide a larger and sustained mortality reduction. PMID- 12081735 TI - New insight into bile flow dynamics in anomalous arrangement of the pancreaticobiliary duct. AB - BACKGROUND: In an anomalous arrangement of the pancreaticobiliary duct (AAPBD), two-way regurgitation, including pancreaticobiliary reflux and biliopancreatic reflux (BPR), might occur. Although pancreaticobiliary reflux has been shown by high levels of pancreatic enzymes in bile, there have been few reports of clinical evidence of BPR in AAPBD. Recently, spiral computed tomography combined with intravenous infusion cholangiography (IVC-SCT) has been reported to be useful for visualizing the biliary anatomy, which may allow a new insight into bile flow dynamics in AAPBD. METHODS: Fifteen paediatric patients with AAPBD were examined before operation using both IVC-SCT and endoscopic retrograde cholangiopancreatography. Visualization of the main pancreatic duct on IVC-SCT defined the presence of BPR. Blood samples were also analysed when the initial symptom was strongly present. RESULTS: BPR was observed in six of 15 patients. Pancreatic enlargement and hyperamylasaemia were significantly more frequent in patients without BPR than in those with BPR. Levels of enzyme markers of cholestasis were also higher in the former group. CONCLUSION: The present study, using IVC-SCT, has identified clinical evidence of BPR in AAPBD. Further studies are necessary to determine the correlation between BPR and clinical manifestations. PMID- 12081736 TI - Transoesophageal echocardiography shows high risk of gas embolism during laparoscopic hepatic resection under carbon dioxide pneumoperitoneum. AB - BACKGROUND: The ultrasonically activated scalpel (UAS) enables safe and effective laparoscopic tissue dissection, making hepatic resection feasible. This study compared blood loss and risk of gas embolism using the UAS during open hepatic resection and laparoscopic hepatic resection. METHODS: Female pigs were divided into two groups for laparoscopic (n = 7) and open (n = 5) left hepatic lobectomy. The UAS was used for both tissue cutting and coagulation. Laparoscopic liver resection was performed under carbon dioxide pneumoperitoneum (intraperitoneal pressure 12 mmHg). During surgery animals were monitored haemodynamically by an arterial line and Swan-Ganz catheter. Two-dimensional transoesophageal echocardiography (2D-TEE) was used to detect gas emboli with special attention to the right atrium and ventricle. Gas emboli were graded according to size, and correlated with haemodynamic and blood gas data. RESULTS: During open and laparoscopic hepatic resection the UAS resulted in minimal blood loss and effective tissue dissection. No air embolism was seen during open surgery. With laparoscopic hepatic resection 2D-TEE revealed gas embolism in all animals. Gas embolism was accompanied by cardiac arrhythmia in four of seven animals. No direct correlation was observed between embolism episodes and blood gas variables. There were no deaths after episodes of embolization. A significant decrease in arterial partial pressure of oxygen was seen at the end of the laparoscopic procedure in all animals. CONCLUSION: The UAS causes minimal blood loss during both open and laparoscopic hepatic resection. Laparoscopic liver dissection under carbon dioxide pneumoperitoneum carries a high risk of gas embolism. PMID- 12081737 TI - Complications and functional outcome following artificial anal sphincter implantation. AB - BACKGROUND: The postoperative complications and functional outcome following 24 consecutive implantations of an artificial anal sphincter were assessed prospectively. METHODS: A total of 24 artificial anal sphincters (Acticon Neosphincter) were implanted in 22 patients (mean age 47 years). The mean follow up period was 28 (range 6-48) months. Results were assessed prospectively by two independent observers at 4-month intervals. The cumulative probability of artificial anal sphincter removal was analysed by the Kaplan-Meier method. RESULTS: Five patients were free of complications. During the postoperative period, complications occurred in nine patients, two of whom required reoperation. During follow-up, complications developed in ten patients, nine of whom were reoperated. Definitive device explantation was necessary in seven patients. The cumulative probability of device explantation was 44 per cent at 48 months. The 15 patients with functioning implants were followed up for a mean of 26 (range 7-48) months. Continence grading improved from a mean of 18 (range 14 20) in the preoperative period to 4 (range 0-14) after operation (P < 0.001). Resting anal canal pressure in patients with a functioning implant increased from a mean of 35 (range 8-87) mmHg before operation to 54 (range 34-70) mmHg after implantation (P < 0.01). CONCLUSION: An artificial anal sphincter is a useful alternative for refractory faecal incontinence but the incidence of late postoperative complications is high. PMID- 12081738 TI - Permanent sacral nerve stimulation for treatment of idiopathic constipation. AB - BACKGROUND: Constipation can usually be managed using conservative therapies. A proportion of patients require more intensive treatment. Surgery provides variable results. This paper describes an alternative approach, in which the neural control of the bowel and pelvic floor is modified, using permanent sacral nerve stimulation. METHODS: Four women (aged 27-36 years), underwent temporary and then permanent stimulation. All had idiopathic constipation, resistant to maximal therapy, with symptoms for 8-32 years. Clinical evaluation, bowel diary, Wexner constipation score, symptom analogue score, quality of life questionnaire and anorectal physiology were completed. RESULTS: There was a marked improvement in all patients with temporary, and in three with permanent, stimulation. Median follow-up was 8 (range 1-11) months. Bowel frequency increased from 1-6 to 6-28 evacuations per 3 weeks. Improvement occurred, at longest-follow-up, in median (range) evacuation score (4 (0-4) versus 1 (0-4)), time with abdominal pain (98 (95-100) versus 12 (0-100) per cent), time with bloating (100 (95-100) versus 12 (5-100) per cent), Wexner score (21 (20-22) versus 9 (1-20)), analogue score (22 (16-32) versus 80 (20-98)) and quality of life. Maximum anal resting and squeeze pressures increased. Rectal sensation was altered. Transit time normalized in one patient. CONCLUSION: Permanent sacral nerve stimulation can be used to treat patients with resistant idiopathic constipation. PMID- 12081739 TI - Retrospective study of acute toxicity following short-course preoperative radiotherapy. AB - BACKGROUND: The use of short-course preoperative radiotherapy (25 Gy in five fractions over 1 week) in resectable rectal cancer reduces local recurrence but is associated with an increased risk of postoperative complications and late toxicity. This study aimed to identify those patients who are unlikely to benefit from short-course preoperative radiotherapy and the factors associated with acute toxicity. METHODS: All patients who received short-course preoperative radiotherapy at a university hospital in 1998 and 1999 were included in this retrospective study. The association between complications occurring within 3 months and patient demographics, radiotherapy technique, surgical details and overall treatment time (OTT) was assessed by univariate and multivariate analysis. RESULTS: The mortality rate at 30 days was 6 per cent in the 177 patients identified. Thirty-seven per cent of patients had either Dukes' A tumours, surgically incurable disease or positive circumferential margins. One or more complications occurred in 38 per cent of patients. On multivariate analysis an OTT of more than 13 days (P = 0.03), age (P = 0.02) and length of the radiotherapy field (P = 0.05) were associated with an increased risk of complications. CONCLUSION: Surgery within 1 week of completing short-course preoperative radiotherapy improved preoperative staging and use of an optimal radiotherapy technique will result in fewer patients at risk of acute toxicity. PMID- 12081740 TI - Medium-term results of permanent sacral nerve stimulation for faecal incontinence. AB - BACKGROUND: Anal sphincter surgery for faecal incontinence is associated with significant morbidity and a variable outcome. Sacral nerve stimulation may provide a good functional outcome with minimal morbidity. This paper reports the experience in a single centre over 5 years. METHODS: Fifteen consecutive patients (14 women), median age 60 (range 37-71) years, underwent temporary, and subsequent permanent, stimulation. All had incontinence to solid or liquid stool; the aetiology was obstetric injury (seven patients), scleroderma (four), idiopathic (two), fistula surgery (one) and repaired rectal prolapse (one). Median duration of symptoms was 6 (range 2-15) years. Clinical evaluation, endoanal ultrasonography, bowel diary, quality of life questionnaire (Short Form (SF) 36) and anorectal physiological testing were performed before and after stimulation. RESULTS: Continence had improved in all patients at median follow-up of 24 (range 3-60) months. Eleven patients were fully continent. Episodes of faecal incontinence decreased from median (range) 11 (2-30) per week before stimulation to 0 (0-4) per week after permanent stimulation (P < 0.001). Urgency improved in all patients (median (range) ability to defer less than 1 (0-1) versus 8 (1-15) min; P = 0.01). 'Social function' and 'role-physical' subscales of the SF36 improved significantly. Mean resting pressure (35 versus 49 cmH2O with temporary stimulation; P < 0.05) and squeeze pressure increment (43 versus 69 cmH2O with permanent stimulation; P < 0.01) increased. Rectal sensitivity to initial distension changed (mean 47 versus 34 ml air; P < 0.05). There were no major complications. CONCLUSION: Sacral nerve stimulation is a safe and effective treatment for faecal incontinence when conventional treatment has failed. There is minimal morbidity. The benefit is maintained in the medium term. PMID- 12081741 TI - Consequences of axillary recurrence after conservative breast surgery. AB - BACKGROUND: The aim was to study the incidence, time course and prognosis of patients who developed axillary recurrence after breast-conserving surgery, and to evaluate possible risk factors for axillary recurrence and prognostic factors after axillary recurrence. METHODS: In a population-based cohort of 6613 women with invasive breast cancer who had breast-conserving surgery between 1981 and 1990, 92 recurrences in the ipsilateral axilla were identified. Risk factors for axillary recurrence were studied in a case-control study nested in the cohort, and late survival was documented in the women with axillary recurrence. RESULTS: The overall risk of axillary recurrence was 1.0 per cent at 5 years and 1.7 per cent at 10 years. The risk of axillary recurrence increased with tumour size (P = 0.033) and was highest in younger women (odds ratio (OR) 3.9 for women aged less than 40 years compared with those aged 50-59 years). Radiotherapy to the breast reduced the risk of axillary recurrence (OR 0.1 (95 per cent confidence interval 0.1 to 0.4)). The breast cancer-specific survival rate after axillary recurrence, as measured from primary treatment, was 78.0 per cent at 5 years and 52.3 per cent at 10 years. Tumour size and node status had a statistically significant effect on death from breast cancer. CONCLUSION: Axillary recurrence is rare, although more common in younger women with large tumours. Radiotherapy to the breast was protective. Tumour size and node status were the most important prognostic factors in women with axillary recurrence. PMID- 12081742 TI - Prognostic factors after oesophagectomy and extended lymphadenectomy for squamous oesophageal cancer. AB - BACKGROUND: The prognosis of patients without nodal metastasis of oesophageal cancer is generally good, but recurrence develops in some cases. METHODS: Data on 88 consecutive patients with squamous oesophageal cancer who underwent three field lymph node dissection from 1986 to September 1998 and who had no evidence of nodal disease were reviewed retrospectively. Disease status was based on histological examination of the section of each node with the largest surface area, stained with haematoxylin and eosin. RESULTS: The 3- and 5-year survival rates of patients without lymph node metastasis were 85 and 81 per cent respectively, better than in patients with metastasis. Twelve patients died from recurrence. Recurrence was haematogenous in nine patients and locoregional in three. Survival was worse in men, for patients with lesions located in the upper thoracic oesophagus, and in those with lymphatic or blood vessel invasion. Only the presence of lymphatic invasion correlated with survival on multivariate analysis (P = 0.04). CONCLUSION: Although survival was generally good in patients without nodal metastasis from oesophageal cancer following three-field lymph node dissection, patients with lymphatic invasion remained at risk for haematogenous dissemination. PMID- 12081743 TI - Cohort study in South and West England of the influence of specialization on the management and outcome of patients with oesophageal and gastric cancers. AB - BACKGROUND: To evaluate specialization in National Health Service (NHS) cancer care, volume-outcome relationships were examined. METHODS: This was a cohort study of 1512 patients with oesophageal or gastric cancer in 23 acute NHS hospitals. Outcomes were survival time and operative (30 day) mortality. Multiple regression analysis was performed, adjusted for diagnoses, prognoses and treatments. RESULTS: For oesophageal cancer, the operative mortality rate decreased by 40 per cent (odds ratio 0.60 (95 per cent confidence interval (c.i.) 0.36 to 0.99 per cent); P = 0.047) for each increase of ten patients in doctors' annual surgical caseloads, and the risk of death decreased by 8 per cent (hazard ratio 0.92 (95 per cent c.i. 0.85 to 0.99); P = 0.021) for each increase of ten patients in doctors' annual caseloads. For gastric cancer, the operative mortality rate decreased by 41 per cent (odds ratio 0.59 (95 per cent c.i. 0.32 to 1.07)) for each increase of ten patients in doctors' annual surgical caseloads, and the risk of death decreased by 7 per cent (hazard ratio 0.93 (95 per cent c.i. 0.89 to 0.98); P = 0.009) for each increase of ten patients in hospitals' annual caseloads. Patients of higher-volume doctors were more likely to receive most investigations and treatments, independently of presenting features. CONCLUSION: The study supports concentration of services for oesophageal and gastric cancers. Specialization of doctors and their teams is at least as important as specialization of hospitals. PMID- 12081744 TI - Revascularization of the superior mesenteric artery after acute thromboembolic occlusion. AB - BACKGROUND: The outcome and prognostic factors after revascularization of acute thromboembolic occlusion of the superior mesenteric artery (SMA) are poorly documented. METHODS: Sixty patients with acute thromboembolic occlusion of the SMA had revascularization procedures at 21 hospitals from 1987 to 1998. They were registered prospectively in the Swedish Vascular Registry. Patient files were analysed retrospectively. RESULTS: The median age of the patients was 76 years; 73 per cent suffered from cardiac disease and 23 per cent had previous vascular surgery. Onset of symptoms was classified as sudden (30 per cent), acute (33 per cent) or insidious (37 per cent). The occlusions were thought to be either embolic (67 per cent) or thrombotic (33 per cent). The diagnosis was suspected on first examination in 32 per cent of patients, a group whose median time to operation was shorter (P = 0.01). Fifty-eight patients had an exploratory laparotomy and subsequent revascularization, and two were treated with thrombolysis alone. Second-look laparotomy was performed in 41, and third look in eight patients; 19 required an additional bowel resection. The overall mortality rates were 43, 52, 60 and 67 per cent at 30 days, discharge, 1 and 5 years, respectively. No patient was dependent on intravenous nutrition after 1 year. Previous vascular surgery resulted in a higher institutional mortality rate (79 per cent; P = 0.02). Patients who had a sudden onset of symptoms outside hospital had a better outcome (mortality rate 27 per cent; P = 0.02). CONCLUSION: Many non diagnostic radiological examinations were performed and a routine second-look is warranted. The results suggest that attempts at revascularization procedures for acute mesenteric ischaemia may improve the outcome. PMID- 12081745 TI - Experimental study of the material composition of laparoscopic ports on tumour cell adherence. AB - BACKGROUND: Laparoscopic resection of intra-abdominal malignancies has yet to be widely adopted, partly because of concerns over the possible increase in the rate of port-site metastasis. The aetiology of these is unclear, but the laparoscopic instrumentation used may influence the deposition of tumour cells at the port sites during operation. An in vitro model to examine tumour cell adherence to laparoscopic ports and to port sites was developed to examine this hypothesis. METHODS: A pilot study (study 1) was performed in which six smooth plastic, six ribbed plastic and six metal ports were introduced through the shaved abdominal wall of a cadaveric sheep and suspended in a water-bath containing radiolabelled LIM 1215 human colonic cancer cells for 30 min. Radioactivity on both ports and port sites was measured and the number of cells adherent to each structure was calculated. The study was expanded to include a further 36 smooth plastic ports and 36 metal ports (study 2). RESULTS: In study 1 metal ports were found to have significantly more adherent cells than plastic ports (P = 0.004), as did ribbed ports when compared with smooth ports (P < 0.05). In study 2 increased numbers of cells were again detected on metal ports (P < 0.001) when compared with plastic ports. Significantly greater numbers of cells were also detected on the sites through which metal ports had passed than on sites through which plastic ports had passed (P = 0.03). CONCLUSION: In this model, the use of metal ports as opposed to plastic ports resulted in increased deposition of tumour cells on both ports and port sites. PMID- 12081746 TI - Renal transplant fibrosis correlates with intragraft expression of tissue inhibitor of metalloproteinase messenger RNA. AB - BACKGROUND: Chronic renal allograft nephropathy is characterized by an abnormal accumulation of extracellular matrix proteins in the glomeruli and tubulo interstitium. The aim of this study was to determine the relationship between intragraft expression of the genes controlling the accumulation of extracellular matrix and the development of chronic renal allograft nephropathy in human renal transplants. METHODS: Forty renal allografts with stable renal function were biopsied 6 months after transplantation. Single glomeruli were plucked from the surface of these protocol biopsies and total messenger RNA (mRNA) was extracted. Reverse transcriptase-polymerase chain reaction was used to study the intragraft expression of several fibrosis-associated genes (collagen III, collagen IValpha2, matrix metalloproteinase (MMP) 2, tissue inhibitors of metalloproteinases (TIMPs) 1 and 2, tenascin and transforming growth factor (TGF) beta1). The level of tubulo-interstitial fibrosis was measured by quantitative immunostaining of collagen III. RESULTS: There were positive correlations between the level of tubulo-interstitial collagen III immunostaining and intragraft expression of the genes for TIMP-1 (rs= 0.70, P < 0.02) and TIMP-2 (rs = 0.59, P < 0.02). Interstitial fibrosis was also strongly correlated with the levels of TGF-beta mRNA (rs = 0.67, P < 0.002). Finally, TIMP-1 expression increased with TGF-beta expression (rs = 0.77, P < 0.002). CONCLUSION: Failure of extracellular matrix degradation may be an important molecular mechanism in the pathogenesis of chronic renal allograft damage. PMID- 12081747 TI - Role of the neutrophil in the development of systemic inflammatory response syndrome and sepsis following abdominal aortic surgery (Br J Surg 2001; 88: 1583 9). PMID- 12081748 TI - Role of the neutrophil in the development of systemic inflammatory response syndrome and sepsis following abdominal aortic surgery (Br J Surg 2001; 88: 1583 9). PMID- 12081750 TI - Cytoreduction including total gastrectomy for pseudomyxoma peritonei (Br J Surg 2002; 89: 208-12). PMID- 12081752 TI - Liver fibrosis increases the risk of intrahepatic recurrence after hepatectomy for hepatocellular carcinoma (Br J Surg 2002; 89: 57-62). PMID- 12081756 TI - The aetiology, pathogenesis and management of priapism. PMID- 12081757 TI - The management of stress urinary incontinence after radical prostatectomy. AB - Up to 30% of patients complain about urine leakage after radical prostatectomy, but persistent stress incontinence (beyond 1 year) affects <5% of them. This complication is mainly caused by sphincter dysfunction. Some preventive measures have been described to decrease the risk of incontinence after radical prostatectomy, but with conflicting results. The effectiveness of preoperative and early postoperative physiotherapy is controversial. Moreover, while meticulous apical dissection of the prostate significantly improves postoperative continence, the benefit of other surgical techniques, e.g. preserving the bladder neck and the neurovascular bundles, is under debate. The treatment of persistent stress urinary incontinence is mainly based on surgery, as this type of incontinence usually does not respond to physiotherapy and anticholinergic medication. While injection therapy is safe and well tolerated, its effect on postoperative continence is limited and decreases with time. The best results are achieved by implanting an artificial urinary sphincter, but with significant complication and revision rates. PMID- 12081758 TI - Patterns and trends in prostate cancer incidence, survival, prevalence and mortality. Part I: international comparisons. AB - The international patterns and trends in prostate cancer incidence, survival, prevalence and mortality were examined. Age-standardized incidence and death rates among men in a variety of countries worldwide were obtained from various sources, survival rates from European sources and elsewhere, and prevalence estimates from the EUROPREVAL study. Results from many published studies were summarized. The incidence of prostate cancer varies widely around the world, with by far the highest rates in the USA and Canada. There has been a gradual increase in the incidence of prostate cancer since the 1960s in many countries and in most continents; there were large increases in the late 1980s and early 1990s in the USA, but increases have also occurred in countries with comparatively low incidence, e.g. India. Survival from prostate cancer improved during the 1970s and 1980s; further increases in the 1990s may be largely a result of earlier diagnosis. There were wide differences in survival across Europe, with rates in the UK well below the average, but all European rates were far below those in the USA. There was wide variation in the prevalence of prostate cancer in Europe; in some countries with high incidence and high life-expectancy, prostate cancers formed approximately 15% of all prevalent cancers in men. Mortality from prostate cancer has also increased in many countries, but to a lesser extent than incidence; this is consistent with the observed trends in survival. Mortality decreased slightly in the mid to late 1990s in several countries, including the USA, Canada, England, France and Austria. Part of the apparent increases in the incidence of prostate cancer has been associated with diagnostic artefacts (particularly detecting preclinical tumours through the increased use of transurethral resection) which may also have had an effect on death certification through the incorrect attribution of prostate cancer as the underlying cause of death. However, the greatest effect on the registration of new cases of prostate cancer has been the increased availability of prostate specific antigen testing during the early- to mid-1990s. Possibly, in addition to the effect of attribution bias, the earlier diagnosis of prostate cancers has contributed to the recent slight decreases in mortality. However, this is unlikely to account for much of the reduction, given the slow development of the disease from onset to death. Changes in disease management are probably more important. There are many strong arguments against introducing population-based screening for prostate cancer. PMID- 12081759 TI - Patterns and trends in prostate cancer incidence, survival, prevalence and mortality. Part II: individual countries. PMID- 12081760 TI - The 'top 10' urological procedures: a study of hospital episodes statistics 1998 99. AB - OBJECTIVE: To summarize the urological procedures recorded in Hospital Episodes Statistics (HES, 1998-99) and to examine the accuracy of coding and HES. MATERIALS AND METHODS: Data on the 10 commonest urological procedures from the Department of Health website (www.doh.gov.uk/hes) were extracted, summarized and presented. RESULTS: Urethral catheterization, endoscopic procedures on the bladder, prostate, urethra and ureter (excluding ureteric stone extraction), minor open procedures on the foreskin and the vas, bladder instillation, extracorporeal shockwave lithotripsy and prostatic biopsy are the 10 commonest procedures, according to finished consultant episodes. There is published evidence that the data from coding and HES are not completely accurate. CONCLUSION: This study highlights shortfalls in the HES data (1998-99) which may directly affect the funding of urological services. PMID- 12081762 TI - Clinical experience with a new ultrasonic and LithoClast combination for percutaneous litholapaxy. AB - OBJECTIVE: To assess a new lithotripter for intracorporal lithotripsy, which combines the mechanically driven pneumatic LithoClast (Electro-Medical Systems, Nyon, Switzerland) and a new ultrasonic device (Electro-Medical Systems), for use in percutaneous nephrolitholapaxy (PNL). PATIENTS AND METHODS: The new lithotripter consists of a LithoClast Master with 12 Hz repetition rate and a new ultrasonic device. The 1.0 mm LithoClast probe is advanced off-centre through the hollow 3.3 mm ultrasonic probe and protrudes about 1 mm. A new irrigation system with a pinch valve compressing the irrigation tube, a foot-switch for activating the ultrasound, the LithoClast and both together, and a stone bucket at the outlet tube are new features. Between February 1999 and August 2001, 68 patients were treated by PNL with the new device; 35 had complete and 33 had partial staghorn calculi. PNL was administered under fluoroscopic control and with the patient prone. RESULTS: The mean (range) duration of surgery was 61 (42-119) min. The complete stone-free rate was 66% after the first PNL; of the 68 patients, 16 received a second PNL, giving a final stone-free rate of 76% and 80%. The stone was composed of calcium oxalate monohydrate (COM) in 13%, COM with uric acid in 35%, apatite in 20% and cystine in 11%. Clinically the new lithotripter was very effective, producing smaller stone particles and thus fewer residual stone fragments after PNL than with the LithoClast or ultrasonic fragmentation alone. CONCLUSION: The new lithotripter provides easily managed and highly effective stone fragmentation of all stones, regardless of their composition. PMID- 12081761 TI - Changing treatments for primary urolithiasis: impact on services and renal preservation in 16,679 patients in Western Australia. AB - OBJECTIVES: To examine changing treatments for the primary presentation of urinary lithiasis and their effects on re-admissions, repeat procedures, cumulative hospital use and renal preservation. PATIENTS AND METHODS: Linked hospital morbidity records were used to identify first-time admissions for renal and ureteric calculi from 1980 to 1997 in the population of Western Australia. The cases were followed to mid-1999 and actuarial methods used to estimate risks of further hospital admissions and procedures, including the loss of a renal moiety. RESULTS: Between 1980 and 1997 the total rate of inpatient procedures for urinary stones more than doubled, at a time when the rate of first-time hospital admissions increased by only 13% and the conservative management of stones remained constant at approximately 59%. The predominant procedure for stone management was initially open lithotomy, replaced in the early 1980s by percutaneous nephrolithotomy and soon supplemented by extracorporeal shock wave lithotripsy (ESWL). The changes in technology led to a four-fold increase in procedural re-admissions within 30 days of primary separation. This was a result of repeated, staged or postponed interventions, often involving the use of stents or a second treatment with ESWL. The risk of surgical intervention decreased from 48% to 32%, whilst the cumulative length of stay over the first year decreased from 7.8 to 3.9 days. The risk of kidney loss declined significantly from 2% to <0.1% during the period. CONCLUSIONS: The main reason for more interventions were short-term procedural re-admissions. ESWL reduced the need for invasive procedures and decreased cumulative hospital stay, despite more re-admissions. Renal preservation improved by a factor of 10. PMID- 12081763 TI - Prognostic significance of nuclear morphometry in renal cell carcinoma. AB - OBJECTIVE: To assess nuclear morphometry as a predictor of prognosis in patients with renal cell carcinoma (RCC). PATIENTS AND METHODS: The study included 65 consecutive patients with RCC who underwent radical nephrectomy and were followed up for a median (range) of 80 (27-138) months. Nuclear morphometry was assessed using a computer-assisted image analysis system on histological sections and characterized by five nuclear variables (area, perimeter, major and minor diameter, and form factor). From the patients' records and pathology specimens, the clinicopathological prognostic variables (histological type, Fuhrman grade and pathological stage) were recorded. The proliferative activity was assessed using immunohistochemical staining with Ki-67 antibody. RESULTS: Higher values of mean nuclear area, perimeter, and major and minor diameter were significantly related to higher nuclear grade, proliferative activity and advanced tumour stage. They were significant predictors of disease progression and survival, together with grade, stage, sarcomatoid histology and proliferative activity. Of all significant prognostic factors predicting progression-free survival, only stage was independent (T4 vs T1, hazard ratio 6.55, 95% CI 1.63-26.13, P=0.008). CONCLUSION: Although the significance of these preliminary results must not be overstated, nuclear morphometry might provide significant prognostic information in predicting survival and tumours at high risk of progression in RCC. PMID- 12081764 TI - Contribution of grade, vascular invasion and age to outcome in clinically localized renal cell carcinoma. AB - OBJECTIVE: To determine the relative prognostic importance of microvascular invasion in apparently localized renal cell carcinoma (RCC). PATIENTS AND METHODS: A retrospective clinical and pathological review was conducted of 176 consecutive patients identified from pathology records who had a nephrectomy for RCC with a median follow-up of 44 months. Vascular invasion was recorded and categorized by the level of microvascular invasion (MVI), renal vein invasion (RVI) and inferior vena cava invasion (IVCI). Tumour type, grade and size were also assessed. These variables were assessed by univariate and multivariate analysis to determine their effect on disease-free survival. RESULTS: In the univariate analysis tumour size, grade, vascular invasion and young age each predicted reduced disease-free survival. On multivariate analysis for all 176 patients, grade, vascular invasion and young age were the significant independent predictors of reduced disease-free survival. In a subgroup of 149 patients from whom those with very high risk determinants were excluded (those with grade 4 tumours and/or IVCI) most of the risk of metastasis could be accounted for by vascular invasion and young age alone (MVI vs no vascular invasion, hazard ratio 3.18, 95% confidence interval 1.29-7.84; RVI vs no vascular invasion 2.41, 0.989 5.89; and age per year 0.963, 0.94-0.992). CONCLUSIONS: Grade, vascular invasion and young age are the main independent predictors of relapse in clinically localized RCC after nephrectomy. For most patients, who do not have very high risk indicators, the main adverse predictors are vascular invasion and young age. These findings are important when selecting patients for trials of adjuvant therapy and have implications for pathological staging. PMID- 12081765 TI - Gender as a prognostic factor in patients with renal cell carcinoma. AB - OBJECTIVE: To evaluate gender as a prognostic factor in patients with renal cell carcinoma (RCC), using a retrospective review of patients with RCC stratified according to gender and analysing factors affecting prognosis. PATIENTS AND METHODS: From January 1957 to December 1995, 768 patients with pathologically defined RCC (all of whom underwent nephrectomy) were classified as having clear cell carcinoma in 662 (follow-up in 648), papillary RCC in 43 (follow-up in 42), chromophobe cell carcinoma in 36 (follow-up in 34) and cyst-associated RCC in 27 (all followed up) according to the criteria proposed by the World Health Organization. The survival rates were compared between men and women, calculated and stratified according to the subtype of RCC. RESULTS: There tended to be a more favourable prognosis in women than in men but the difference was not quite significant (P=0.061). Of those with clear cell carcinoma, women had a more favourable prognosis than men and the difference in survival was significant (P=0.012). No other subtype of RCC was associated with a significant difference in survival between the sexes. There was a smaller proportion of patients with stage IV and a larger proportion with stage I disease in women than in men (P<0.05). Of stage I patients, women had a more favourable prognosis than men (P<0.011). Women had better survival after recurrence than had men, the difference being significant (P=0.007). CONCLUSION: The prognosis is significantly better in women than men with clear cell carcinoma. The factors that contribute to a favourable prognosis in women are the greater proportion of lower stage disease and better survival after recurrence. PMID- 12081766 TI - Reliability and validity of the Malay version of the Health-Related Quality of Life instrument in a Malaysian population. AB - OBJECTIVE: To validate the Malay version of the Health-Related Quality of Life (Mal-HRQOL-20) questionnaire in patients with and without urinary symptoms in a Malaysian population. PATIENTS AND METHODS: The validity and reliability of the Mal-HRQOL-20 were assessed in patients with and without lower urinary tract symptoms (LUTS). The reliability was evaluated using the test-retest method and the internal consistency using Cronbach's alpha. Sensitivity to change was expressed as the effect size in the score before and after intervention in additional patients with LUTS who underwent transurethral resection of the prostate. RESULTS: The internal consistency was excellent; there was a high degree of internal consistency for each of the 20 items and for the overall score (Cronbach's alpha > or = 0.57 and 0.79, respectively) in the population study. The test-retest correlation coefficient for the 20 item scores was highly significant. The intra-class correlation coefficient was high (> or = 0.55). The sensitivity and specificity were high for the effects of treatment. There was a very significant agreement between scores before and after treatment across all domains in the treatment cohort, but not in the control group. CONCLUSION: The Mal-HRQOL-20 is suitable, reliable, valid and sensitive to clinical change in the Malaysian population. PMID- 12081767 TI - Alternative medications for benign prostatic hyperplasia available on the Internet: a review of the evidence for their use. PMID- 12081768 TI - Nurse-led cystoscopy: the next step. AB - OBJECTIVES: To determine and compare the accuracy of flexible cystoscopy and subsequent management by a specialist urology registrar (SpR) and a specialist urology nurse (SUN). PATIENTS AND METHODS: Flexible cystoscopy was undertaken on 50 patients by both endoscopists each unaware of the results of the other's examination. The findings and management plans were independently declared and then compared. Any cystoscopic discrepancies were assessed by one consultant with the patients under a general anaesthetic. RESULTS: Tumours were found in 20% of patients; there was a consensus of the endoscopic findings and subsequent management plan between the SpR and the SUN in 94% of the patients. Any missed tumours (all surveillance) were minute and clinically insignificant. The chance corrected proportional agreement (kappa value) between the SUN and final diagnosis and management was 0.94. CONCLUSION: A properly trained SUN can undertake both diagnostic and surveillance flexible cystoscopy, and decide upon further management to the same degree as can a urology SpR. PMID- 12081769 TI - Ileal T-pouch as a urinary continent cutaneous diversion: clinical and urodynamic evaluation. AB - OBJECTIVE: To report the functional results of continent cutaneous urinary diversion with ileum, using the serous-lined extramural valve and Mitrofanoff procedure for the continence mechanism. PATIENTS AND METHODS: From April 1999 to October 2001, 18 patients (mean age 68 years) underwent radical cystectomy for invasive bladder cancer involving the bladder neck, urethra or prostate; they then had an ileal continent cutaneous T pouch constructed. The mean (range) follow-up was 12 (4-20) months. The first five patients had reconstructions using the original orthotopic T-pouch configuration, but in subsequent patients the technique was simplified, reducing the ileal segments. The ureteric-intestinal anastomosis was made using a split-cuff nipple technique. RESULTS: All patients were continent day and night, and there were no late complications. All cutaneous continent T pouches had a good capacity and low pressure, with no urinary reflux. No catheterization difficulties were reported and the evacuation intervals were approximately 4 h. CONCLUSIONS: This ileal cutaneous continent diversion is a versatile technique; the T-valve can be used successfully as a continent mechanism, ensuring continence day and night. The simplified technique maintains a reservoir of good capacity and compliance, thus preserving approximately 13 cm of ileal tract. The ureteric intestinal anastomosis by the split-cuff nipple technique is suitable in undilated and peristaltic ureters. PMID- 12081770 TI - A new core-biopsy instrument with an end-cut technique provides prostate biopsies with increased tissue yield. AB - OBJECTIVE: To evaluate the tissue yield of a new core-biopsy needle with an end cut mechanism when used for transrectal prostate biopsies. PATIENTS AND METHODS: The end-cut instrument has an adjustable stroke length (13, 23 or 33 mm) and an inner and an outer cannula, but no trocar. It was compared with the conventional side-notch needle (Biopty, Bard, Covington, GA, USA) with trocar and fixed stroke length (22 mm). In 60 men, octant biopsies were taken including the apex, mid medial, mid-lateral and basal positions. At random, the left side of the prostate was biopsied with one of the instruments and the right side with the other. The length and weight of the biopsy specimens were measured. In 40 men, the stroke length of the end-cut instrument was set to 23 mm, and in 20 men it was set to 33 mm. RESULTS: The weight and the weight per length of the biopsies provided by the end-cut instrument with a stroke length of 23 mm were 18.4% and 13.7%, respectively, greater than in the biopsies from the side-notch instrument (P<0.001). In 21.9% of the attempts no tissue was obtained by the end-cut needle, compared with 1.9% with the side-notch needle. With a 33 mm stroke length, the length and weight of the end-cut biopsies were 38.4% and 33.0%, respectively, greater than in biopsies from the side-notch instrument. CONCLUSION: The end-cut instrument provided a greater tissue yield than the side-notch needle, but a significant number of biopsies were lost. PMID- 12081771 TI - The prevalence and predictors of psychological distress in patients with early localized prostate cancer. AB - OBJECTIVE: To determine the level of psychopathology, traumatic distress and quality of life in men with newly diagnosed clinically localized prostate cancer, the effect on these of a consultation in a combined-specialist early-prostate clinic, and predictors of psychopathology. PATIENTS AND METHODS: Eighty-eight patients were recruited from the combined clinic; they completed a battery of questionnaires including the Hospital Anxiety and Depression Scale (HADS), the revised Impact of Event Scale (IES) and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire, before their first appointment. Two weeks later they completed the HADS, IES and a patient satisfaction survey. RESULTS: The overall level of psychopathology varied among the questionnaires used, from 0% on the HADS depression scale, 8% on the HADS anxiety scale and 14% on the IES. Anxiety and traumatic stress symptoms were commoner than depressive symptoms. The quality-of-life scores showed a relatively good level of functioning. Pre-morbid factors and disease status did not predict psychological distress. Younger age was mildly predictive of poorer psychological functioning. Anxiety symptoms reduced slightly after a joint clinic appointment, whereas depressive symptoms showed a slight increase. CONCLUSION: This study suggests that men with early localized prostate cancer have low levels of psychopathology overall. However, some men experience distressing psychological symptoms and it is important that future research is conducted to help develop clear guidelines on the optimal methods of detecting and managing men with prostate cancer who have mental health difficulties. PMID- 12081772 TI - Urodynamic findings 3 months after radiotherapy in patients treated with conformal external beam radiotherapy for prostate carcinoma. AB - OBJECTIVE: To quantify the effect of radiotherapy (RT) on urodynamic function 3 months after RT in patients with prostate cancer undergoing definitive external beam RT. PATIENTS AND METHODS: Seventeen patients with clinically localized prostate cancer were accrued into a single-arm prospective study. Sixteen of the patients completed a scheduled multichannel video-urodynamic study at baseline and again 3 months after RT; the urodynamic variables were then compared to assess the nature and extent of urodynamic change caused by RT. Correlations were assessed between these quantitative changes and those in self-assessed qualitative urinary function measured by International Prostate Symptom Score (IPSS), Quality of Life assessment index (QoL) and urinary functional enquiry. RESULTS: There were significant changes detected by the urodynamic study 3 months after RT in bladder volume at capacity (mean decrease 70 mL) and bladder volume at first sensation when supine (mean decrease 85 mL), and a lower postvoid residual volume (mean 50 mL). There was no significant change in the remaining urodynamic variables (including maximum flow rate and voided volume), nor in bladder compliance, bladder instability or bladder outlet obstruction. The self assessed qualitative urological function measured by the IPSS, QoL and median urinary frequency/24 h showed no significant change after RT. CONCLUSIONS: This is the first quantitative study to prospectively evaluate the effect of RT on urodynamics in patients with prostate cancer. Only a few urodynamic variables changed significantly 3 months after RT, while most, including self-assessed qualitative urinary function, did not. This finding corresponds well with the notion that most patients tolerate RT well and that acute RT-induced urinary symptoms resolve successfully, with the return of lower urinary tract function to baseline levels by 3 months after RT. PMID- 12081773 TI - End-to-end urethroplasty: long-term results. AB - OBJECTIVE: To retrospectively review patients with strictures (<3 cm) of the bulbous urethra who had undergone urethroplasty with excision of the stenotic segment and end-to-end anastomosis. PATIENTS AND METHODS: The review included 74 patients (all men, mean age 39 years, range 18-70) treated between 1989 and 1999 for strictures 5-30 mm long. Forty-one of the patients (55%) had been treated previously, 39 endoscopically (urethrotomy and/or dilatation) and two surgically. Surgical access was perineal, with the patient in an exaggerated lithotomy position; the stenotic segment was excised and the stumps spatulated for end-to end anastomosis. The mean (range) duration of surgery was 140 (75-280) min. There were no complications during or after surgery, and none related to the duration in the lithotomy position. RESULTS: At a mean follow-up of 60 months, 93% of the patients had no recurrence of the stricture and were therefore considered cured. There were no treatment-related complications. CONCLUSION: End-to-end anastomosis is confirmed as the treatment of choice for short bulbous urethral strictures, giving cure rates close to 100%. PMID- 12081774 TI - Does delaying pyeloplasty affect renal function in children with a prenatal diagnosis of pelvi-ureteric junction obstruction? AB - OBJECTIVE: To evaluate retrospectively children with an antenatal diagnosis of pelvi-ureteric junction (PUJ) obstruction who required pyeloplasty for deteriorating renal function during the follow-up, specifically assessing the recovery of function after surgery. PATIENTS AND METHODS: Between 1988 and 2000, 44 consecutive patients with a prenatal diagnosis of PUJ obstruction who were initially treated by observation underwent pyeloplasty because their renal function deteriorated during the follow-up. Patients with bilateral disease, a single kidney or vesico-ureteric reflux were excluded from the study. The mean (sd) age of the patients was 13.3 (4.5) months and their mean (range) initial renal function 39.7 (30.4-45)%. In all patients the diagnostic criteria and indications for surgery were identical. The main indication for surgery was deteriorating renal function of >5%, confirmed by renal scintigraphy. Of 44 patients, 35 (77%) had severe dilatation of the renal pelvis by >3 cm. The mean (sd) deterioration in renal function during the follow-up, just before surgery, was 8.2 (2.4)%, and the mean time between the last acceptable renogram and the study showing deterioration was 8.3 (1.3) months; the mean follow-up was 5.1 (1.6) years. The results were assessed statistically using the unpaired nonparametric Mann-Whitney U-test. RESULTS: An improvement in hydronephrosis was confirmed in all patients, with renal function returning to the initial levels in 36 of 44 (81%) patients 6-12 months after surgery, although in two patients renal function did not improve after surgery. There was no significant difference between the preoperative characteristics and the degree of hydronephrosis, renal function or patient age with the level of renal functional improvement after surgery. CONCLUSION: In patients with an antenatal diagnosis of PUJ obstruction, expectant management is recommended and spares the children unnecessary surgery. Even if renal function deteriorates, delayed pyeloplasty recovers the initial functional level. PMID- 12081775 TI - Does monosymptomatic enuresis exist? A molecular genetic exploration of 32 families with enuresis/incontinence. AB - OBJECTIVE: s To confirm linkage to microsatellite markers on chromosome 8q, 12q, 13q and 22q in families with nocturnal enuresis/incontinence segregating with an autosomal dominant pattern, and to determine if there is an association between the clinical subtype and these linked loci. PATIENTS AND METHODS: Families with at least three members with nocturnal enuresis in two generations were included in the study. The index patient was > or = 7 years old and had evidence of bladder dysfunction; all other family members were > or = 5 years old. Bladder dysfunction in the index patients was documented by video-urodynamics when indicated. A nycthemeral rhythm of diuresis was documented in all index patients. The clinical diagnosis of all family members was based on a questionnaire on voiding problems and micturition habits, uroflowmetry, measurement of functional bladder capacity and nocturnal diuresis. Linkage was analysed using an autosomal dominant model with a gene frequency equal to 0.05 and a penetrance of 0.9. RESULTS: Thirty-two families with nocturnal enuresis/incontinence (one with four, 25 with three and six with two generations) were included. The mean number of persons included per family was 10 and on average five members were symptomatic. Linkage of nocturnal enuresis to a region on chromosome 22q11 was found in nine families, to 13q13-14 in six and to 12q in four. There was no convincing evidence for linkage to chromosome 8q. Clinical findings in the proband and their family members with possible linkage to a given locus were heterogeneous, and hence no clear genotype/phenotype correlation could be postulated. CONCLUSION: These findings support the hypothesis of the genetic and phenotypic heterogeneity of nocturnal enuresis/incontinence. Putative linkage was confirmed to the same chromosomal loci as in previous studies of 'monosymptomatic' enuresis and different phenotypes were linked to the same loci. PMID- 12081776 TI - High initial efficacy of full-spectrum therapy for nocturnal enuresis in children and adolescents. AB - OBJECTIVE: To investigate the initial efficacy and predictive factors of full spectrum therapy in the treatment of children and young adolescents with nocturnal enuresis (NE). PATIENTS AND METHODS: Combined therapy for NE comprises an enuresis alarm, bladder training, motivational therapy and pelvic floor muscle training, and is more effective than each of the components alone or than medical intervention. A total of 60 children and adolescents (aged 4-20 years) with NE were treated once a week with full-spectrum therapy for a maximum of 6 months. RESULTS: Overall the therapy was successful (14 consecutive dry nights) in 52 of 60 patients. At 30 days the cure rate was 33%, after 60 days 72% and after 98 days, 87%. The remaining 13% did not achieve 14 consecutive dry nights; seven patients improved, having fewer dry nights/week. One patient discontinued the treatment because of lack of motivation. In children with an initial maximum bladder capacity less than normal for age, the capacity increased from 53% of the normal maximum bladder capacity in week 1 to 88% at the end of treatment. Neither age, gender, sleep arousal, bladder capacity, family history and pathophysiological profile had any association with the success rate. CONCLUSION: The short-term success rate of full-spectrum therapy for NE is high. Age, gender, sleep arousal, bladder capacity, family history and pathophysiological profile of enuresis are unrelated to the success of the intervention. PMID- 12081777 TI - Distal hypospadias: which repair? AB - OBJECTIVE: To compare the function, complications and cosmesis after a modified Thiersch-Duplay and Mathieu unstented urethroplasty. PATIENTS AND METHODS: Over a 5-year period 381 consecutive patients (not randomized) with distal hypospadias were evaluated. A modified Thiersch-Duplay with dorsal incised urethral plate tubularization (group A) was undertaken in 170 (45%) and a parameatal based flip flap Mathieu hypospadias repair (group B) in 211 (55%). No urinary diversion or stent was used in any of the 381 patients. RESULTS: In both groups the mean (sd) follow-up was 3.1 (1.4) years. All 381 patients voided spontaneously after surgery and none developed urinary retention needing catheterization. In groups A and B, respectively, the overall late complications were 12 (7.1%) and 32 (15.2%) (P=0.001), with urethrocutaneous fistula in six (3.5%) and 26 (12.3%; P=0.001); secondary surgery for fistula repair was successful in all boys in A and 89% in B. The glanular meatus was a vertical slit in all in group A and 86% in B (P=0.02). In both groups, 71% of the children who are now toilet-trained and standing to void have a good calibre, single and straight urinary stream in a forward direction. CONCLUSIONS: Stenting or urinary diversion is unnecessary after distal hypospadias surgery. The functional results were good in those standing to void. A more natural vertical slit-like glanular meatus was easily created using the modified Thiersch-Duplay urethroplasty, with a lower fistula rate. PMID- 12081778 TI - Paediatric penile trauma. AB - OBJECTIVE: To report our experience with paediatric penile trauma in a retrospectively evaluated series. PATIENTS AND METHODS: The records of 64 boys (mean age 7 years, sd 4) who were hospitalized over the last 20 years because of penile trauma were reviewed. The cause of trauma was circumcision in 43 (67%), a human hair-tie strangulation injury in 10 (16%), an animal attack in four (6%), a bicycle accident in four (6%), a zipper injury in two (3%) and electrical injury in one (2%). Patients were managed according to the severity of the injury. Eight (12%) with minimal skin loss or meatal injury underwent primary skin closure or meatoplasty; 40 (62%) with urethrocutaneous fistulae underwent repair and five (8%) with a glans hanging on a thin pedicle had the glans and the urethra reconstructed. Patients with partial or complete amputation of the glans (10) underwent primary haemostasis and meatoplasty; the penis was lengthened in one. One child with complete avulsion of the penis underwent perineal urethrostomy. RESULTS: Fifty-four patients (84%) were followed for a mean (sd) of 5.7 (4) years; there were good cosmetic and functional results in 45 (83%). Fifteen patients are now adults; 13 (86%) reported normal sexual function. Of the 40 patients assessed with circumcision-related injuries, six (15%) had functional disability (short penis in one and fistulae in five). Of the 10 patients with a hair-tie injury, none lost their glans. Of the four injuries caused by animal attacks, three had poor results (emasculation in one, short penis in one and severe curvature in the remaining patient). There was no functional disability in the remaining forms of trauma. CONCLUSIONS: In our region, ritual circumcision and hair-tie strangulation injuries are the most common causes of penile trauma in children. Good functional and cosmetic results are possible in most cases. However, animal attacks are associated with the highest rate of long-term functional and cosmetic disability. PMID- 12081779 TI - The presence or absence of an impalpable testis can be predicted from clinical observations alone. AB - OBJECTIVE: To evaluate, in a prospective study, the accuracy of predicting the presence or absence of unilateral or bilateral impalpable testes from a clinical examination, particularly whether the contralateral descended testis (CDT) is hypertrophied. PATIENTS AND METHODS: Whether the ipsilateral scrotal appendages were palpable, and the size of the CDT, were determined before surgery in a series of patients, and compared with age-matched controls. Between 1992 and 2000, 100 impalpable testes in 86 consecutive patients (mean age at orchidopexy 45 months, range 6-223; 66% <36 months) were evaluated and treated. In addition to the presence or absence of palpable ipsilateral scrotal appendages, the size of the CDT, when present, and the intraoperative findings were recorded. Logistic regression analysis was used to model the probability of the presence or absence of the testis, as determined by the preoperative clinical findings alone. RESULTS: Of the 86 testes that were located at surgery, 50 (58%) were intracanalicular, 28 (32%) intra-abdominal and the remainder (10%) were in the superficial inguinal pouch. Of 13 patients with the 'vanishing testis syndrome', the atrophic testicle was intracanalicular in nine, in the upper scrotum in three and intra-abdominal in only one. All viable testes were successfully relocated in the scrotum, with one atrophic after surgery. The positive predictive value (PPV, with 95% confidence interval) of a testis being present when the ipsilateral appendages were palpable and the CDT was not hypertrophied was 0.93 (0.83-0.97). Conversely, the PPV of the impalpable testis being absent when the appendages were impalpable and the CDT was hypertrophied was 0.95 (0.64-0.99). CONCLUSION: When evaluating and surgically treating impalpable testes, the presence of palpable ipsilateral scrotal appendages and a CDT with no hypertrophy is associated with a 93% likelihood of discovering a testis that can be successfully relocated to the scrotum. Conversely, when the ipsilateral scrotal appendages cannot be palpated and the CDT is hypertrophied, there is a 96% probability that the impalpable testis is absent (vanishing testis syndrome). This readily available information may be valuable in preoperative counselling and planning. PMID- 12081780 TI - Diuretic Doppler ultrasonography in chronic unilateral partial ureteric obstruction in dogs. AB - OBJECTIVE: To compare the effects of diuresis induced by a loop diuretic (frusemide), an osmotic diuretic (mannitol) and a high-osmolar radio-contrast medium (sodium iothalamate) on the intrarenal resistive index (RI) in dogs with chronic unilateral partial ureteric obstruction (UPUO). MATERIALS AND METHODS: The split renal clearance and intrarenal RI were estimated in 11 dogs with chronic UPUO. Doppler ultrasonography measurements of the interlobar arteries were obtained before and 10 min after the intravenous infusion of 1 mg/kg frusemide, 1 g/kg mannitol or 20 mL 60% sodium iothalamate. The same experimental protocol was repeated with another drug at 1-week intervals. RESULTS: There was a significant difference between the intrarenal RI of obstructed and unobstructed kidneys in the chronic phase of UPUO. While mannitol and sodium iothalamate significantly increased the RI in both kidneys, differences in RI between the kidneys decreased after infusing the two drugs. However, while frusemide insignificantly increased the RI in the obstructed kidney, it decreased the RI in unobstructed one. Consequently, the difference in RI between the kidneys increased significantly after administering frusemide. There were no significant differences in urinary volume after administering each of the drugs. CONCLUSIONS: As there were no significant differences in the diuretic effects of the drugs, frusemide may have additional effects on the RI of unobstructed kidneys other than diuresis. Frusemide increased the difference between the intrarenal RI of the kidneys and therefore may improve the detection of unilateral urinary obstruction in humans. PMID- 12081781 TI - Localization of peripheral dopamine D1 and D2 receptors in rat corpus cavernosum. AB - OBJECTIVE: To detect and locate anatomically peripheral dopamine D1 and D2 receptors in rat cavernosa, as dopamine is important in sexual drive and penile erection through receptors located in the central nervous system. MATERIALS AND METHODS: Corpora cavernosa were obtained from Sprague-Dawley rats; total RNA and membrane proteins were extracted and cryostat sections prepared. The rat brain hypothalamus was used as a control for dopamine D1 and D2 receptors. The presence and expression of peripheral dopamine D1 and D2 receptor mRNAs in rat corpus cavernosa was assessed using reverse transcription and polymerase chain reaction (RT-PCR), and Northern blot hybridization using (32)P-UTP-labelled RNA probes. Concurrently, corresponding proteins from D1 and D2 receptors were assayed and detected by a Western blotting technique. The anatomical location of dopamine D1 and D2 receptor mRNAs in rat penile tissues was identified by in situ hybridization using (35)S-UTP-labelled RNA probes in cryostat sections. Immunohistochemical staining was used to locate peripheral dopamine D1 and D2 receptor proteins in rat corpora cavernosa. RESULTS: Dopamine D1 and D2 receptor gene expression was detected in rat corpora cavernosa. In situ hybridization signals for dopamine D1 and D2 receptor mRNAs were localized to corpus cavernosal tissues and dorsal vessels in the rat penis. Western blot analyses showed peripheral dopamine D1 and D2 receptor proteins in rat corpora cavernosa. Immunohistochemically, peripheral dopamine D1 and D2 receptor proteins were detected in dorsal nerves, dorsal vessels and corpus cavernosal smooth muscle of the rat penile tissues. CONCLUSIONS: Peripheral dopamine D1 and D2 receptors are present in the corpora cavernosa of rats. The functional significance of these receptors and signal transduction pathways in modulating the vascular tone of the penis warrants further investigation. PMID- 12081783 TI - Gap junctions and connexin expression in human suburothelial interstitial cells. AB - OBJECTIVE: To determine whether suburothelial interstitial cells of the human bladder express gap junctions, and if so, to establish their extent and composition, using immunocytochemistry, confocal microscopy and electron microscopy. MATERIALS AND METHODS: Bladder tissue was obtained at cystectomy; the tissue was: (i) frozen for cryosectioning and Northern blot analysis; (ii) fixed and embedded for standard thin-section electron microscopy; and (iii) processed using low-denaturation conditions in Lowicryl for immunogold-label electron microscopy. Cryosections were immunofluorescently labelled using antibodies against connexins 43, 40 and 45, vimentin, desmin and c-Kit ligand, and examined by confocal microscopy. Double labelling was used to determine the spatial relationship of labelling for connexin43 with that of vimentin and desmin. Thin section electron microscopy was used to investigate interstitial cell ultrastructure and permit unequivocal identification of gap junctions, and immunogold labelling of Lowicryl sections was applied to localize connexin43. RESULTS: Immunoconfocal microscopy showed prominent labelling for the gap junction protein, connexin43, in a suburothelial band of cells that was also strongly positive for vimentin. The connexin43/vimentin-positive cells showed only weak labelling for desmin and c-Kit ligand, and were immunonegative for connexins 40 and 45. Northern blotting showed a corresponding abundance of connexin43 transcript in the mucosal layer but not the detrusor layer of the bladder wall. Electron microscopy revealed abundant gap junctions, recognized by their pentalaminar structure, between the cell processes of interstitial cells in the suburothelial zone. That these interstitial cell gap junctions were the source of the connexin43 immunolabelling observed by immunoconfocal microscopy was confirmed by immunogold labelling in sections of Lowicryl-embedded tissue examined by electron microscopy. CONCLUSION: A network of interstitial cells, extensively linked by connexin43-containing gap junctions, is located beneath the urothelium in human bladder. As gap junctions provide pathways for direct cell-to cell communication, the interstitial cellular network may operate as a functional syncytium, integrating signals and responses in the bladder wall. PMID- 12081782 TI - Antisense Bcl2 oligonucleotide in cisplatin-resistant bladder cancer cell lines. AB - OBJECTIVE: To determine the change of expression of Bcl2 in cisplatin-resistant bladder cancer cell lines and the reversibility of chemoresistance to cisplatin with antisense oligonucleotide against Bcl2, as higher expression of Bcl2 is associated with drug resistance in many different cancer cell lines. MATERIALS AND METHODS: In the cisplatin-resistant bladder tumour cell lines T24R1 and T24R2, the expression of Bcl2 was determined by reverse transcription-polymerase chain reaction and Western blot assay, and antisense oligonucleotide targeting of the Bcl2 coding sequence was administered with lipofectin. RESULTS: The expression of Bcl2 mRNA and protein was greater in T24R1 and T24R2 cells than in the parent T24 cells. Short-term exposure to cisplatin up-regulated Bcl2 mRNA and protein expression in parent T24 cells. Treatment with antisense oligonucleotide down-regulated Bcl2 protein expression and significantly enhanced the cytotoxicity of cisplatin. CONCLUSIONS: Up-regulation of Bcl2 protein expression might be one of the mechanisms of cisplatin resistance in bladder cancer cells, and antisense Bcl2 oligonucleotide may be helpful in chemotherapy for bladder cancer by reversing cisplatin resistance. PMID- 12081784 TI - Congenital variations in mucomuscular development of the ureter. PMID- 12081785 TI - Misplacement of an intrauterine contraceptive device into the ureter after uterine wall perforation. PMID- 12081786 TI - Percutaneous cryoablation of renal cell carcinoma in a transplanted kidney. PMID- 12081788 TI - Benign lateralizing haematuria: the impact of upper tract endoscopy. PMID- 12081790 TI - Can diet affect prostate cancer? PMID- 12081791 TI - Can diet affect prostate cancer? PMID- 12081794 TI - Laparoscopy in urology: indications and training. PMID- 12081797 TI - Laparoscopic lymph-node sampling in locally advanced prostate cancer. PMID- 12081799 TI - A review on SNP and other types of molecular markers and their use in animal genetics. AB - During the last ten years, the use of molecular markers, revealing polymorphism at the DNA level, has been playing an increasing part in animal genetics studies. Amongst others, the microsatellite DNA marker has been the most widely used, due to its easy use by simple PCR, followed by a denaturing gel electrophoresis for allele size determination, and to the high degree of information provided by its large number of alleles per locus. Despite this, a new marker type, named SNP, for Single Nucleotide Polymorphism, is now on the scene and has gained high popularity, even though it is only a bi-allelic type of marker. In this review, we will discuss the reasons for this apparent step backwards, and the pertinence of the use of SNPs in animal genetics, in comparison with other marker types. PMID- 12081800 TI - Prediction error variance and expected response to selection, when selection is based on the best predictor - for Gaussian and threshold characters, traits following a Poisson mixed model and survival traits. AB - In this paper, we consider selection based on the best predictor of animal additive genetic values in Gaussian linear mixed models, threshold models, Poisson mixed models, and log normal frailty models for survival data (including models with time-dependent covariates with associated fixed or random effects). In the different models, expressions are given (when these can be found - otherwise unbiased estimates are given) for prediction error variance, accuracy of selection and expected response to selection on the additive genetic scale and on the observed scale. The expressions given for non Gaussian traits are generalisations of the well-known formulas for Gaussian traits - and reflect, for Poisson mixed models and frailty models for survival data, the hierarchal structure of the models. In general the ratio of the additive genetic variance to the total variance in the Gaussian part of the model (heritability on the normally distributed level of the model) or a generalised version of heritability plays a central role in these formulas. PMID- 12081802 TI - A Bayesian approach for constructing genetic maps when markers are miscoded. AB - The advent of molecular markers has created opportunities for a better understanding of quantitative inheritance and for developing novel strategies for genetic improvement of agricultural species, using information on quantitative trait loci (QTL). A QTL analysis relies on accurate genetic marker maps. At present, most statistical methods used for map construction ignore the fact that molecular data may be read with error. Often, however, there is ambiguity about some marker genotypes. A Bayesian MCMC approach for inferences about a genetic marker map when random miscoding of genotypes occurs is presented, and simulated and real data sets are analyzed. The results suggest that unless there is strong reason to believe that genotypes are ascertained without error, the proposed approach provides more reliable inference on the genetic map. PMID- 12081803 TI - Results of a whole genome scan targeting QTL for growth and carcass traits in a Pietrain x Large White intercross. AB - We herein report the results of a whole genome scan performed in a Pietrain x Large White intercross counting 525 offspring to map QTL influencing economically important growth and carcass traits. We report experiment-wide significant lod scores (>4.6) for meatiness and fat deposition on chromosome SSC2, and for average daily gain and carcass length on chromosome SSC7. Additional suggestive lod scores (>3.3) for fat deposition are reported on chromosomes SSC1, SSC7 and SSC13. A significant dominance deviation was found for the QTL on SSC1, while the hypothesis of an additive QTL could not be rejected for the QTL on SSC7 and SSC13. No evidence for imprinted QTL could be found for QTL other than the one previously reported on SSC2. PMID- 12081801 TI - Use of maternal information for QTL detection in a (grand)daughter design. AB - In a (grand)daughter design, maternal information is often neglected because the number of progeny per dam is limited. The number of dams per maternal grandsire (MGS), however, could be large enough to contribute to QTL detection. But dams and MGS usually are not genotyped, there are two recombination opportunities between the MGS and the progeny, and at a given location, only half the progeny receive a MGS chromosomal segment. A 3-step procedure was developed to estimate: (1) the marker phenotypes probabilities of the MGS; (2) the probability of each possible MGS haplotype; (3) the probabilities that the progeny receives either the first, or second MGS segment, or a maternal grandam segment. These probabilities were used for QTL detection in a linear model including the effects of sire, MGS, paternal QTL, MGS QTL and maternal grandam QTL. Including the grandam QTL effect makes it possible to detect QTL in the grandam population, even when MGS are not informative. The detection power, studied by simulation, was rather high, provided that MGS family size was greater than 50. Using maternal information in the French dairy cattle granddaughter design made it possible to detect 23 additional QTL genomewise significant. PMID- 12081804 TI - Cytogenetic and molecular characterization of eight new reciprocal translocations in the pig species. Estimation of their incidence in French populations. AB - Eight new cases of reciprocal translocation in the domestic pig are described. All the rearrangements were highlighted using GTG banding techniques. Chromosome painting experiments were also carried out to confirm the proposed hypotheses and to accurately locate the breakpoints. Three translocations, rcp(4;6)(q21;p14), rcp(2;6)(p17;q27) and rcp(5;17)(p12;q13) were found in boars siring small litters (8.3 and 7.4 piglets born alive per litter, on average, for translocations 2/6 and 5/17, respectively). The remaining five, rcp(5;8)(p12;q21), rcp(15;17)(q24;q21), rcp(7;8)(q24;p21), rcp(5;8)(p11;p23) and rcp(3;15)(q27;q13) were identified in young boars controlled before entering reproduction. A decrease in prolificacy of 22% was estimated for the 3/15 translocation after reproduction of the boar carrier. A parental origin by inheritance of the translocation was established for the (5;8)(p11;p23) translocation. The overall incidence of reciprocal translocations in the French pig populations over the 2000/2001 period was estimated (0.34%). PMID- 12081805 TI - A method to optimize selection on multiple identified quantitative trait loci. AB - A mathematical approach was developed to model and optimize selection on multiple known quantitative trait loci (QTL) and polygenic estimated breeding values in order to maximize a weighted sum of responses to selection over multiple generations. The model allows for linkage between QTL with multiple alleles and arbitrary genetic effects, including dominance, epistasis, and gametic imprinting. Gametic phase disequilibrium between the QTL and between the QTL and polygenes is modeled but polygenic variance is assumed constant. Breeding programs with discrete generations, differential selection of males and females and random mating of selected parents are modeled. Polygenic EBV obtained from best linear unbiased prediction models can be accommodated. The problem was formulated as a multiple-stage optimal control problem and an iterative approach was developed for its solution. The method can be used to develop and evaluate optimal strategies for selection on multiple QTL for a wide range of situations and genetic models. PMID- 12081806 TI - Optimal selection on two quantitative trait loci with linkage. AB - A mathematical approach to optimize selection on multiple quantitative trait loci (QTL) and an estimate of residual polygenic effects was applied to selection on two linked or unlinked additive QTL. Strategies to maximize total or cumulative discounted response over ten generations were compared to standard QTL selection on the sum of breeding values for the QTL and an estimated breeding value for polygenes, and to phenotypic selection. Optimal selection resulted in greater response to selection than standard QTL or phenotypic selection. Tight linkage between the QTL (recombination rate 0.05) resulted in a slightly lower response for standard QTL and phenotypic selection but in a greater response for optimal selection. Optimal selection capitalized on linkage by emphasizing selection on favorable haplotypes. When the objective was to maximize total response after ten generations and QTL were unlinked, optimal selection increased QTL frequencies to fixation in a near linear manner. When starting frequencies were equal for the two QTL, equal emphasis was given to each QTL, regardless of the difference in effects of the QTL and regardless of the linkage, but the emphasis given to each of the two QTL was not additive. These results demonstrate the ability of optimal selection to capitalize on information on the complex genetic basis of quantitative traits that is forthcoming. PMID- 12081807 TI - A further look at quantitative trait loci affecting growth and fatness in a cross between Meishan and Large White pig populations. AB - A detailed quantitative trait locus (QTL) analysis of growth and fatness data from a three generation experimental cross between Large White (LW) and Meishan (MS) pig breeds was carried out to search for sex x QTL interactions, imprinting effects and multiple linked QTLs. A total of 530 F(2) males and 573 F(2) females issued from 6 F(1) boars and 23 F(1) sows were typed for a total of 137 markers covering the entire porcine genome. Nine growth traits and three backfat thickness measurements were analysed. All analyses were performed using line cross regression procedures. A QTL with sex-specific expression was revealed in the proximal region of chromosome 8, although some confusion between herd and sex effects could not be discarded. This previously undetected QTL affected male growth during the fattening period, with a favourable additive effect of the LW allele. The analyses also revealed the presence of two linked QTLs segregating on chromosome 1, affecting growth traits during the post-weaning period. The first QTL, previously detected using a single QTL model, was located at the end of the q arm of chromosome 1 and had a favourable MS allele. The second QTL had a favourable LW allele and was located in the proximal extremity of the q arm of chromosome 1. Suggestive genomic imprinting was found in the distal region of chromosome 9 affecting growth during the fattening period. PMID- 12081808 TI - Heritability of susceptibility to Salmonella enteritidis infection in fowls and test of the role of the chromosome carrying the NRAMP1 gene. AB - 373 thirteen-week-old chicks issued from a commercial cross and 312 chickens from the L2 line were intravenously inoculated with 10 (6) Salmonella enteritidis and the numbers of Salmonella in the spleen, liver and genital organs were assessed 3 days later. Heritabilities of the number of Salmonella were estimated at 0.02 +/ 0.04 and 0.05 +/-0.05 in the liver; at 0.29 +/-0.07 and 0.10 +/-0.06 in the spleen; and at 0.16 +/- 0.05 and 0.11 +/-0.08 in the genital organs, in the first and second experiments, respectively. The difference between the two experiments could result from sampling variations and from differences in the genetic structure of the two populations possibly including both heterosis and additive effects as well as their interaction in the first experiment. Genetic correlations between the number of bacteria in the genital organs and liver (0.56 +/-0.58 and 0.76 +/- 0.32 in the first and second experiments, respectively) and spleen (0.37 +/-0.24 and 0.79 +/-0.23) were positive. Moreover a significant within-sire effect of VIL1, a marker gene for NRAMP1, was observed in 117 progeny resulting from 25 informative matings. These results indicate that there are genetic differences in the resistance to visceral infection by S. enteritidis in these commercial egg-laying flocks, and suggest that these differences are at least partly due to genetic polymorphism in the NRAMP1 region. PMID- 12081809 TI - Muscle-related traits in cattle: The role of the myostatin gene in the South Devon breed. AB - In this paper, we examined the effects of an 11-bp mutation within the GDF-8 gene, originally identified in Belgian Blue cattle, in the South Devon breed. The mutation was found at moderate frequency (0.37) in the South Devon population. We quantified the effects of this mutation on growth, body composition and calving traits in South Devon cattle. We found that the mutation significantly increased muscle score and calving difficulty and decreased fat depth. The mutation did not significantly affect weight at 200 and 400 days or muscle depth. Its effect on muscle score and fat depth was additive while its effect on calving difficulty was recessive. The mutation accounted for a significant proportion of the phenotypic variance in muscle score and calving difficulty. There was an economic benefit of the mutation for this data set, however, calculations were sensitive to changes in the parameter values. Additional data would be required to refine these calculations. PMID- 12081810 TI - Microsatellite loci in Japanese quail and cross-species amplification in chicken and guinea fowl. AB - In line with the Gifu University's initiative to map the Japanese quail genome, a total of 100 Japanese quail microsatellite markers isolated in our laboratory were evaluated in a population of 20 unrelated quails randomly sampled from a colony of wild quail origin. Ninety-eight markers were polymorphic with an average of 3.7 alleles per locus and a mean heterozygosity of 0.423. To determine the utility of these markers for comparative genome mapping in Phasianidae, cross species amplification of all the markers was tested with chicken and guinea fowl DNA. Amplification products similar in size to the orthologous loci in quail were observed in 42 loci in chicken and 20 loci in guinea fowl. Of the cross-reactive markers, 57.1% in chicken and 55.0% in guinea fowl were polymorphic when tested in 20 birds from their respective populations. Five of 15 markers that could cross-amplify Japanese quail, chicken, and guinea fowl DNA were polymorphic in all three species. Amplification of orthologous loci was confirmed by sequencing 10 loci each from chicken and guinea fowl and comparing with them the corresponding quail sequence. The microsatellite markers reported would serve as a useful resource base for genetic mapping in quail and comparative mapping in Phasianidae. PMID- 12081811 TI - A higher resolution radiation hybrid map of bovine chromosome 13. AB - In this paper, we present a radiation hybrid framework map of BTA13 composed of nine microsatellite loci, six genes and one EST. The map has been developed using a recently constructed 12'000 rad bovine-hamster whole-genome radiation hybrid panel. Moreover, we present a comprehensive map of BTA13 comprising 72 loci, of which 45 are microsatellites, 20 are genes and seven are ESTs. The map has an estimated length of 2694.7 cR(12'000). The proposed order is in general agreement with published maps of BTA13. Our results only partially support previously published information of five blocks of conserved gene order between cattle and man. We found no evidence for the existence of an HSA20 homologous segment of coding DNA on BTA13 located centromeric of a confirmed HSA10 homologous region. The present map increases the marker density and the marker resolution on BTA13 and enables further insight into the evolutionary development of the chromosome as compared to man. PMID- 12081813 TI - Fruit and vegetables--building a solid foundation. PMID- 12081814 TI - Too little vitamin D in premenopausal women: why should we care? PMID- 12081815 TI - International table of glycemic index and glycemic load values: 2002. AB - Reliable tables of glycemic index (GI) compiled from the scientific literature are instrumental in improving the quality of research examining the relation between GI, glycemic load, and health. The GI has proven to be a more useful nutritional concept than is the chemical classification of carbohydrate (as simple or complex, as sugars or starches, or as available or unavailable), permitting new insights into the relation between the physiologic effects of carbohydrate-rich foods and health. Several prospective observational studies have shown that the chronic consumption of a diet with a high glycemic load (GI x dietary carbohydrate content) is independently associated with an increased risk of developing type 2 diabetes, cardiovascular disease, and certain cancers. This revised table contains almost 3 times the number of foods listed in the original table (first published in this Journal in 1995) and contains nearly 1300 data entries derived from published and unpublished verified sources, representing > 750 different types of foods tested with the use of standard methods. The revised table also lists the glycemic load associated with the consumption of specified serving sizes of different foods. PMID- 12081816 TI - A single daily dose of soybean phytosterols in ground beef decreases serum total cholesterol and LDL cholesterol in young, mildly hypercholesterolemic men. AB - BACKGROUND: Consumption of phytosterol-supplemented margarine lowers total plasma cholesterol (TC) and LDL-cholesterol concentrations in older middle-aged hypercholesterolemic individuals. The effects of incorporating phytosterols into lower-fat foods on the plasma lipids of young men at increased risk of developing cardiovascular disease have not been studied. OBJECTIVE: We tested the hypothesis that a single daily dose of soybean phytosterols added to ground beef will lower plasma TC and LDL-cholesterol concentrations in mildly hypercholesterolemic young men. DESIGN: In a triple-blind, 4-wk study, 34 male college students with elevated plasma TC (5.85 +/- 0.70 mmol/L), LDL cholesterol (4.02 +/- 0.60 mmol/L), and TC:HDL cholesterol (5.5 +/- 1.2) were randomly assigned to the control (ground beef alone) or treatment (ground beef with 2.7 g of phytosterols) group. The phytosterol mixture was two-thirds esterified and one-third nonesterified and consisted of beta-sitosterol (48%), campesterol (27%), and stigmasterol (21%). RESULTS: Consumption of phytosterol-supplemented ground beef lowered plasma TC and LDL-cholesterol concentrations and TC:HDL cholesterol from baseline by 9.3%, 14.6%, and 9.1%, respectively (P < 0.001). The LDL particle size did not change, suggesting that the decrease was primarily of particle number. The decreases were similar in subjects with (n = 8) and without (n = 9) a family history of premature cardiovascular disease. No significant changes were found in the control group. CONCLUSION: Phytosterol-supplemented ground beef effectively lowers plasma TC and LDL cholesterol and has the potential to become a functional food to help reduce the risk of cardiovascular disease. PMID- 12081817 TI - Breath acetone is a reliable indicator of ketosis in adults consuming ketogenic meals. AB - BACKGROUND: Ketogenic diets are used therapeutically to treat intractable seizures. Clinically, it appears that the maintenance of ketosis is crucial to the efficacy of the diet in ameliorating seizures. To understand how ketosis and seizure protection are related, a reliable, noninvasive measure of ketosis that can be performed frequently with minimal discomfort is needed. OBJECTIVE: The objective was to determine which index, breath acetone or urinary acetoacetate, is more strongly related to the plasma ketones acetoacetate and beta hydroxybutyrate. DESIGN: After fasting overnight for 12 h, 12 healthy adults consumed 4 ketogenic meals over 12 h. Blood, breath, and urine samples were collected hourly. Blood was analyzed for plasma acetoacetate and beta hydroxybutyrate, breath for acetone, and urine for acetoacetate. RESULTS: By the end of the 12-h dietary treatment, plasma acetoacetate, plasma beta hydroxybutyrate, and breath acetone had increased 3.5-fold, whereas urinary acetoacetate increased 13-fold when measured enzymatically and 25-fold when measured with urinary ketone dipsticks. Plasma acetoacetate was best predicted by breath acetone (R(2) = 0.70, P < 0.0001). Plasma beta-hydroxybutyrate was equally predicted by breath acetone and urinary acetoacetate (R(2) = 0.54, P = 0.0040). CONCLUSIONS: Breath acetone is as good a predictor of ketosis as is urinary acetoacetate. Breath acetone analysis is noninvasive and can be performed frequently with minimal discomfort to patients. As an indicator of ketosis in epilepsy patients consuming a ketogenic diet, breath acetone may be useful for understanding the mechanism of the diet, elucidating the importance of ketosis in seizure protection, and ultimately, enhancing the efficacy of the diet by improving patient monitoring. PMID- 12081818 TI - Timing of vagal stimulation affects postprandial lipid metabolism in humans. AB - BACKGROUND: Vagal stimulation combined with an oral fat load enhances postprandial lipemia in animals and humans. OBJECTIVE: We assessed whether the observed postprandial increase in plasma lipids could be explained by changes in exogenous (chylomicron) or endogenous (VLDL) lipid metabolism and whether the timing of vagal stimulation in relation to fat intake was important. DESIGN: Vagal stimulation was achieved by using the modified sham feeding (MSF) technique, in which food is tasted and chewed but not swallowed. Seven healthy men consumed an oral fat load (50 g) on one occasion (control protocol). On 2 other occasions, they consumed an oral fat load combined with MSF of an appetizing meal. MSF was performed for either 1 h before or 1 h after the oral fat load. Blood was collected for 7 h and was analyzed for hormones and metabolites. RESULTS: The postprandial triacylglycerol response differed significantly (P < 0.001) between the 3 protocols. Both MSF studies resulted in significantly higher plasma pancreatic polypeptide concentrations compared with the control. Compared with MSF for 1 hour after fat intake, MSF for 1 h before fat intake resulted in significantly higher plasma insulin concentrations (P = 0.013), a more rapid rise in chylomicron triacylglycerol concentrations (P = 0.04), and higher VLDL triacylglycerol and apoliprotein B-100 concentrations. CONCLUSIONS: Vagal stimulation enhanced postprandial lipemia via effects on both chylomicron and VLDL metabolism. MSF before fat intake had more dramatic effects on postprandial lipemia than did MSF after fat intake, possibly because of increased parasympathetic activity at the time of ingestion. PMID- 12081819 TI - A comparison of the effects of 2 doses of soy protein or casein on serum lipids, serum lipoproteins, and plasma total homocysteine in hypercholesterolemic subjects. AB - BACKGROUND: Studies have shown that soy protein reduces some atherogenic lipid and lipoprotein concentrations, although lipoprotein(a) concentrations may be increased. The dose response of soy protein has not been established; neither has its effect on plasma total homocysteine. OBJECTIVE: Our objective was to evaluate the effect of 2 doses of soy protein on lipid, lipoprotein, and homocysteine concentrations. DESIGN: Four to 24 wk after being instructed to consume a lipid lowering diet, 130 men and women with LDL-cholesterol concentrations > or = 4 mmol/L were studied during a parallel group trial in which 4 interventions were assigned randomly. Thirty grams isolated soy protein (ISP) and 10 g cotyledon fiber or 50 g ISP and 16.6 g cotyledon fiber or equivalent doses of casein and cellulose were consumed daily as a beverage for 16 wk. RESULTS: When the 2 groups who consumed ISP were compared with the 2 groups who consumed casein, the differences in the net changes from baseline to week 16 in the concentrations of LDL cholesterol and plasma total homocysteine were -0.26 mmol/L (95% CI: -0.43, 0.09 mmol/L; P = 0.01) and -0.8 micromol/L (-1.4, -0.2 micromol/L; P = 0.005), respectively. The effect of the ISP dose was not significant. There were no significant differences between the 2 ISP and the 2 casein groups in changes in lipoprotein(a), HDL-cholesterol, or triacylglycerol concentrations. CONCLUSIONS: Adding 30-50 g soy protein/d to a lipid-lowering diet significantly reduced LDL cholesterol concentrations without increasing lipoprotein(a) concentrations. Plasma total homocysteine concentrations also decreased, suggesting a novel, possibly antiatherosclerotic effect. PMID- 12081820 TI - Cardiovascular disease risk factors and n-3 fatty acid status in the adult population of James Bay Cree. AB - BACKGROUND: Canadian native populations, which traditionally consume large amounts of fish, have lower rates of mortality from heart disease than do Canadian nonnative populations, which have low fish intakes. Fish oils rich in n 3 fatty acids may have a protective effect against cardiovascular disease (CVD) risk factors. OBJECTIVES: The purposes were to examine the profile of plasma phospholipid concentrations of the n-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) among James Bay Cree and to verify the relation between these concentrations and CVD risk factors. DESIGN: The study population consisted of 917 subjects aged 18-74 y who participated in the 1991 Sante Quebec Health Survey. Data were obtained through home interviews and clinic visits. Plasma samples were analyzed for phospholipid fatty acid composition. RESULTS: The mean fish consumption on the day before the survey was 60 g among the adult Cree population. Expressed as a percentage of total fatty acids, relative concentrations of EPA and DHA were 0.65% and 2.80%, respectively. n-3 Fatty acids were higher among coastal residents than among inland residents. A positive association was observed between plasma HDL and n-3 fatty acids. EPA and EPA+DHA were inversely associated with triacylglycerols. Among subjects aged 50-74 y, an inverse association between EPA and EPA:AA and total:HDL cholesterol was observed. CONCLUSIONS: n-3 Fatty acids may favorably influence some CVD risk factors. The Cree population must be encouraged to maintain their traditional fish-based diet, which may be one of the factors protecting them against mortality from CVD. PMID- 12081821 TI - Fruit and vegetable intake and risk of cardiovascular disease in US adults: the first National Health and Nutrition Examination Survey Epidemiologic Follow-up Study. AB - BACKGROUND: Epidemiologic studies report inconsistent findings on the association of fruit and vegetable intake with the risk of cardiovascular disease. OBJECTIVE: The objective was to examine the relation between fruit and vegetable intake and the risk of cardiovascular disease. DESIGN: We studied 9608 adults aged 25-74 y participating in the first National Health and Nutrition Examination Survey Epidemiologic Follow-up Study and free of cardiovascular disease at the time of their baseline examination between 1971 and 1975. Fruit and vegetable intake at baseline was measured with a food-frequency questionnaire. The incidence of and mortality from cardiovascular disease were obtained from medical records and death certificates. RESULTS: Over an average of 19 y, 888 strokes (218 fatal), 1786 ischemic heart disease events (639 fatal), 1145 cardiovascular disease deaths, and 2530 all-cause deaths were documented. Consuming fruit and vegetables > or = 3 times/d compared with <1 time/d was associated with a 27% lower stroke incidence [relative risk (RR): 0.73; 95% CI: 0.57, 0.95; P for trend = 0.01), a 42% lower stroke mortality (0.58; 0.33, 1.02; P for trend = 0.05), a 24% lower ischemic heart disease mortality (0.76; 0.56, 1.03; P for trend = 0.07), a 27% lower cardiovascular disease mortality (0.73; 0.58, 0.92; P for trend = 0.008), and a 15% lower all-cause mortality (0.85; 0.72, 1.00; P for trend = 0.02) after adjustment for established cardiovascular disease risk factors. CONCLUSION: We showed an inverse association of fruit and vegetable intake with the risk of cardiovascular disease and all-cause mortality in the general US population. PMID- 12081823 TI - Food intake and meal patterns of weight-stable and weight-gaining persons. AB - BACKGROUND: Weight gain is a result of changes in the regulation of short-term meal-to-meal intake. An investigation of the short-term intake and activity levels of weight-gaining persons may provide insight into the nature of the cues signaling weight gain. OBJECTIVE: The basic hypothesis was that the investigation of energy balance during periods of dynamic weight gain should provide clues to the regulatory differences that result in obesity. DESIGN: The eating behavior and activity levels of 19 weight-gaining men and women and of weight-stable, matched control subjects were compared with the use of 7-d diet diaries. Participants recorded their activity levels, everything that they ate or drank, and the environmental and psychological factors surrounding each eating episode for 7 consecutive days. RESULTS: The weight-gaining group ingested 1645 kJ/d more than did the weight-stable group because of a greater consumption of carbohydrate and fat and larger meal sizes. CONCLUSION: The greater food intake in the weight gaining group did not result from environmental, social, or psychological factors, suggesting that the overeating associated with weight gain might be physiologically based. PMID- 12081822 TI - Dietary intake and nutritional status of young vegans and omnivores in Sweden. AB - BACKGROUND: Adolescents sometimes become vegetarian for ethical rather than health reasons. This may result in health problems caused by lack of interest in and knowledge of nutrition. OBJECTIVE: We compared the dietary intake and nutritional status of young Swedish vegans and omnivores. DESIGN: The dietary intakes of 30 vegans (15 males and 15 females; mean age: 17.5 +/- 1.0 y) and 30 sex-, age-, and height-matched omnivores were assessed with the use of a diet history interview and validated by the doubly labeled water method and by measuring nitrogen, sodium, and potassium excretion in urine. Iron status and serum vitamin B-12 and folate concentrations were measured in blood samples. RESULTS: The diet-history method underestimated energy intake by 13% and potassium intake by 7% compared with the doubly labeled water method and 24-h urine excretion, respectively. Reported dietary nitrogen and sodium intakes agreed with the 24-h urinary excretion measure. Vegans had higher intakes of vegetables, legumes, and dietary supplements and lower intakes of cake and cookies and candy and chocolate than did omnivores. Vegans had dietary intakes lower than the average requirements of riboflavin, vitamin B-12, vitamin D, calcium, and selenium. Intakes of calcium and selenium remained low even with the inclusion of dietary supplements. There was no significant difference in the prevalence of low iron status among vegans (20%) and omnivores (23%). Two vegans with low intakes of vitamin B-12 had low serum concentrations. CONCLUSION: The dietary habits of the vegans varied considerably and did not comply with the average requirements for some essential nutrients. PMID- 12081825 TI - Carbohydrate administration during a day of sustained aerobic activity improves vigilance, as assessed by a novel ambulatory monitoring device, and mood. AB - BACKGROUND: The brain requires a continuous supply of glucose to function adequately. During aerobic exercise, peripheral glucose requirements increase and carbohydrate supplementation improves physical performance. The brain's utilization of glucose also increases during aerobic exercise. However, the effects of energy supplementation on cognitive function during sustained aerobic exercise are not well characterized. OBJECTIVE: The effects of energy supplementation, as liquid carbohydrate, on cognitive function during sustained aerobic activity were examined. DESIGN: A double-blind, placebo-controlled, between-subjects design was used. Young, healthy men (n = 143) were randomly assigned to 1 of 3 treatment groups. The groups received either a 6% (by vol) carbohydrate (35.1 kJ/kg), 12% (by vol) carbohydrate (70.2 kJ/kg), or placebo beverage in 6 isovolumic doses, and all groups consumed 2 meals (3200 kJ). Over the 10-h study, the subjects performed physically demanding tasks, including a 19.3-km road march and two 4.8-km runs, interspersed with rest and other activities. Wrist-worn vigilance monitors, which emitted auditory stimuli (20/h) to which the subjects responded as rapidly as possible, and a standardized self report mood questionnaire were used to assess cognitive function. RESULTS: Vigilance consistently improved with supplemental carbohydrates in a dose-related manner; the 12% carbohydrate group performed the best and the placebo group the worst (P < 0.001). Mood-questionnaire results corroborated the results from the monitors; the subjects who received carbohydrates reported less confusion (P = 0.040) and greater vigor (P = 0.025) than did those who received the placebo. CONCLUSIONS: Supplemental carbohydrate beverages enhance vigilance and mood during sustained physical activity and interspersed rest. In addition, ambulatory monitoring devices can continuously assess the effects of nutritional factors on cognition as individuals conduct their daily activities or participate in experiments. PMID- 12081824 TI - Muscle fiber type IIX atrophy is involved in the loss of fat-free mass in chronic obstructive pulmonary disease. AB - BACKGROUND: Although the loss of peripheral muscle mass has been shown convincingly in chronic obstructive pulmonary disease (COPD), the underlying pathogenesis remains unclear. OBJECTIVE: The aim of the present study was to determine the relations between skeletal muscle fiber types, fiber cross sectional area (CSA), enzyme activities, and fat-free mass (FFM) in patients with COPD and in control subjects. DESIGN: In 15 patients with COPD and 15 healthy, age-matched control subjects, FFM was determined by dual-energy X-ray absorptiometry and bioelectrical impedance analysis. In biopsy specimens from the vastus lateralis fiber types, fiber CSA and activities of cytochrome oxidase (EC 1.9.3.1), succinate dehydrogenase (EC 1.3.99.1), and glycogen phosphorylase (EC 2.4.1.1) were examined immunohistochemically and histochemically. RESULTS: Compared with control subjects, patients with COPD had less FFM (49 compared with 59 kg, P = 0.030) and lower mean fiber CSA (3839 compared with 4647 microm(2), P = 0.037). A strong correlation (r = 0.87, P < 0.001) was observed between the FFM measured by bioelectrical impedance analysis and mean fiber CSA in patients with COPD. Within fiber-type categories the mean CSA of only the IIA/IIX and IIX fiber types was lower in patients than in control subjects [3358 compared with 4428 microm(2) (P = 0.022) and 2566 compared with 4248 microm(2) (P = 0.003), respectively]. In COPD, 20% of the type IIX fibers lacked stainable activities of cytochrome oxidase, succinate dehydrogenase, and glycogen phosphorylase, and this proportion correlated negatively with type IIX fiber CSA (r = -0.65, P = 0.012). CONCLUSIONS: Muscle fiber atrophy occurs in the vastus lateralis in patients with COPD and contributes to the loss of muscle mass in COPD. Atrophy is specific to fiber types IIA/IIX and IIX and is associated with a disturbed metabolic capacity. PMID- 12081826 TI - Overexpression of arginase I in enterocytes of transgenic mice elicits a selective arginine deficiency and affects skin, muscle, and lymphoid development. AB - BACKGROUND: Arginine is required for the detoxification of ammonia and the synthesis of proteins, nitric oxide, agmatine, creatine, and polyamines, and it may promote lymphocyte function. In suckling mammals, arginine is synthesized in the enterocytes of the small intestine, but this capacity is lost after weaning. OBJECTIVE: We investigated the significance of intestinal arginine production for neonatal development in a murine model of chronic arginine deficiency. DESIGN: Two lines of transgenic mice that express different levels of arginase I in their enterocytes were analyzed. RESULTS: Both lines suffer from a selective but quantitatively different reduction in circulating arginine concentration. The degree of arginine deficiency correlated with the degree of retardation of hair and muscle growth and with the development of the lymphoid tissue, in particular Peyer's patches. Expression of arginase in all enterocytes was necessary to elicit this phenotype. Phenotypic abnormalities were reversed by daily injections of arginine but not of creatine. The expression level of the very arginine-rich skin protein trichohyalin was not affected in transgenic mice. Finally, nitric oxide synthase-deficient mice did not show any of the features of arginine deficiency. CONCLUSIONS: Enterocytes are important for maintaining arginine homeostasis in neonatal mice. Graded arginine deficiency causes graded impairment of skin, muscle, and lymphoid development. The effects of arginine deficiency are not mediated by impaired synthesis of creatine or by incomplete charging of arginyl-transfer RNA. PMID- 12081827 TI - The effect of etomoxir on 24-h substrate oxidation and satiety in humans. AB - BACKGROUND: The carnitine O-palmitoyltransferase I (EC 2.3.1.21) inhibitor etomoxir inhibits fatty acid oxidation, and hepatic fatty acid oxidation has been suggested to be a metabolic satiety signal in subjects who consume high-fat diets. OBJECTIVE: We investigated substrate oxidation and satiety after repeated administrations of etomoxir or placebo in subjects who consumed a high-fat diet. DESIGN: In a randomized crossover design consisting of three 5-d treatments, we fed 10 healthy men [mean +/- SE age: 25.6 +/- 1.7 y; mean +/- SE body mass index (in kg/m(2)): 21.8 +/- 0.3] a high-fat diet twice and a low-fat diet once. The subjects consumed each diet at home for 3 consecutive days, after which they spent 36 h in energy balance in a respiration chamber. During the chamber stays with the high-fat treatments, etomoxir or placebo was administered in 5 doses (600 mg etomoxir in total). Blood samples were obtained on the mornings of days 4 and 5 of each treatment, and appetite profiles were assessed. RESULTS: Mean (+/ SE) 24-h respiratory quotients were significantly (P < 0.05) higher with repeated administrations of etomoxir (0.833 +/- 0.004) than with repeated administrations of placebo (0.814 +/- 0.006), and mean (+/-SE) 24-h whole-body fat oxidation tended to be less (13.7%, P = 0.06) with administration of etomoxir (136.0 +/- 5.2 g/d) than with administration of placebo (157.5 +/- 5.6 g/d). With the etomoxir treatment, fat balance was positive (P < 0.0001) and carbohydrate balance was negative (P < 0.001), whereas with the placebo treatment, neither of the balances was significantly different from zero. Hunger and satiety ratings were not affected under these conditions. CONCLUSIONS: Etomoxir decreased whole body fat oxidation, as indicated by the respiratory quotients in the healthy subjects. With the current protocol, however, hunger and satiety ratings were not affected. PMID- 12081828 TI - Glucose and insulin responses to dietary chromium supplements: a meta-analysis. AB - BACKGROUND: Several authors, mostly on the basis of nonrandomized studies, have suggested dietary trivalent chromium supplementation as an attractive option for the management of type 2 diabetes and for glycemic control in persons at high risk of type 2 diabetes. OBJECTIVE: The study aimed to determine the effect of chromium on glucose and insulin responses in healthy subjects and in individuals with glucose intolerance or type 2 diabetes. DESIGN: The study design was a systematic review and meta-analysis of randomized clinical trials (RCTs). RESULTS: The authors identified 20 reports of RCTs assessing the effect of chromium on glucose, insulin, or glycated hemoglobin (Hb A(1c)). This review summarizes data on 618 participants from the 15 trials that reported adequate data: 193 participants had type 2 diabetes and 425 were in good health or had impaired glucose tolerance. The meta-analysis showed no association between chromium and glucose or insulin concentrations among nondiabetic subjects. A study of 155 diabetic subjects in China showed that chromium reduced glucose and insulin concentrations; the combined data from the 38 diabetic subjects in the other studies did not. Three trials reported data on Hb A(1c): one study each of persons with type 2 diabetes, persons with impaired glucose tolerance, and healthy subjects. The study of diabetic subjects in China was the only one to report that chromium significantly reduced Hb A(1c). CONCLUSIONS: Data from RCTs show no effect of chromium on glucose or insulin concentrations in nondiabetic subjects. The data for persons with diabetes are inconclusive. RCTs in well characterized, at-risk populations are necessary to determine the effects of chromium on glucose, insulin, and Hb A(1c). PMID- 12081829 TI - Diet and iron status of nonpregnant women in rural Central Mexico. AB - BACKGROUND: Few studies have examined the relation of iron status to diet in populations from developing countries with high levels of iron deficiency and diets of poor quality. OBJECTIVE: The objective was to identify nutrients, dietary constituents, and foods that are associated with better iron status in a rural Mexican population. DESIGN: A prospective cohort study was conducted in rural central Mexico. The subjects were 125 nonpregnant women aged 16-44 y. During the 12 mo before blood collection, food intakes were assessed repeatedly by a combination of dietary recalls, food weighing, and food diaries [mean (+/ SD) days of food intake data: 18.8 +/- 5.9 d]. Hemoglobin, hematocrit, and plasma ferritin were measured at the end of the study. RESULTS: Higher plasma ferritin concentrations were associated with greater intakes of nonheme iron and ascorbic acid after control for age, BMI, breast-feeding, season, and the time since the birth of the last child. Higher ascorbic acid intakes, but not higher intakes of heme and nonheme iron, predicted a lower risk of low hemoglobin and hematocrit values after control for the background variables. Consumption of the alcoholic beverage pulque predicted a lower risk of low ferritin and low hemoglobin values. Seasonal variation in ferritin, hemoglobin, and hematocrit values was observed. CONCLUSION: Better iron status was associated with greater intakes of foods containing nonheme iron and ascorbic acid. PULQUE:a beverage containing iron, ascorbic acid, and alcohol-may influence the iron status of women in rural central Mexico. PMID- 12081830 TI - Iron indexes and total antioxidant status in response to soy protein intake in perimenopausal women. AB - BACKGROUND: Elevated iron stores, oxidative stress, and estrogen deficiency may place postmenopausal women at greater risk of heart disease and cancer than premenopausal women. OBJECTIVE: The objective was to determine the effect of soy protein isolate (SPI) intake and iron indexes on plasma total antioxidant status (TAS) in perimenopausal women after control for other contributing factors. DESIGN: Perimenopausal women (n = 69) were randomly assigned (double blind) to treatment: isoflavone-rich SPI (SPI+; n = 24), isoflavone-poor SPI (SPI-; n = 24), or whey protein (control; n = 21). Each subject consumed 40 g soy or whey protein daily for 24 wk. Plasma TAS, serum ferritin, serum iron, transferrin saturation, and hemoglobin were measured at baseline, week 12, and week 24. RESULTS: No significant time-by-treatment interactions on iron indexes or TAS were observed, whereas time had an effect on serum ferritin (P < or = 0.0001) and hemoglobin (P = 0.004) but not on TAS. Multiple regression analysis showed that at week 12, 48% (P < or = 0.0001) of the variability in TAS was accounted for by baseline TAS, alcohol intake, soy intake (soy compared with control; P = 0.016), plasma lipoprotein(a), and dietary iron. At week 24, 47% of the variability in TAS was accounted for by baseline TAS, serum ferritin, serum estrone, dietary zinc, and dietary meat, fish, and poultry. CONCLUSIONS: SPI intake had no significant effect on iron status, but our results suggest that dietary soy protein and low iron stores may protect perimenopausal women from oxidative stress. PMID- 12081831 TI - Individual carotenoid concentrations in adipose tissue and plasma as biomarkers of dietary intake. AB - BACKGROUND: Plasma and adipose tissue concentrations of carotenoids are thought to reflect short- and long-term intakes of carotenoids, respectively. The ability of adipose tissue carotenoid concentrations to reflect dietary intake in population studies is unknown. OBJECTIVE: We examined the relation between intakes of the major dietary carotenoids and their concentrations in plasma and adipose tissue. DESIGN: A blood sample and an adipose tissue biopsy sample were collected from 115 women and 344 men in Costa Rica after they had fasted overnight, and a dietary interview based on a 135-item food-frequency questionnaire was administered. After carotenoid intake was adjusted for total energy intake and plasma concentrations were adjusted for HDL-, LDL-, and VLDL cholesterol concentrations, we calculated partial Spearman correlation coefficients that were adjusted for age, sex, smoking, and body mass index. RESULTS: In women, the correlations (r) between intakes and concentrations of alpha-carotene, beta-carotene, beta-cryptoxanthin, and lutein+zeaxanthin were 0.25, 0.29, 0.44, and 0.17, respectively (P < 0.05 for r > or = 0.19), in adipose tissue and 0.26, 0.13, 0.55, and 0.22 in plasma. In men, these values were 0.04, 0.07, 0.23, and 0.06 in adipose tissue and 0.24, 0.22, 0.44, and 0.20 in plasma. In women and men, correlations for lycopene were higher in plasma (r = 0.19 and 0.35, respectively) than in adipose tissue (r = 0.14 and 0.26). The relative abundance of each carotenoid in the diet was similar to its distribution in plasma but not in adipose tissue. CONCLUSION: The usefulness of adipose tissue and plasma carotenoids as biomarkers of intake is similar, although correlations for individual carotenoids vary substantially. PMID- 12081832 TI - Methylenetetrahydrofolate reductase 677C-->T genotype modulates homocysteine responses to a folate-rich diet or a low-dose folic acid supplement: a randomized controlled trial. AB - BACKGROUND: Low folate status and elevated plasma homocysteine are associated with increased risk of neural tube defects and cardiovascular disease. Homocysteine responses to folate may be influenced by genetic variants in folate metabolism. OBJECTIVE: We determined the effect of folate-enhancing dietary interventions on plasma folate and plasma total homocysteine (tHcy) with respect to the methylenetetrahydrofolate reductase 677C-->T genotype. DESIGN: A total of 126 healthy subjects (42 TT, 42 CT, and 42 CC genotypes) completed 3 dietary interventions (4 mo each) in random order: 1) exclusion diet (avoidance of folic acid-fortified foods and ingestion of a placebo daily), 2) folate-rich diet (increased intake of fortified and naturally folate-rich foods to achieve 400 microg folate/d), and 3) supplement (exclusion diet plus a folate supplement of 400 microg/d). RESULTS: Plasma folate was higher (P < or = 0.001) and plasma tHcy lower (P < or = 0.001) after the folate-rich and supplement interventions than after the exclusion diet. Plasma folate was significantly greater after supplementation than after the folate-rich diet, but there was no significant difference in tHcy concentration (P = 0.72). TT homozygotes had higher plasma tHcy (14.5 compared with 8.9 micromol/L, P < or = 0.001) and lower plasma folate (14.8 compared with 19.0 nmol/L, P < or = 0.01) than did subjects with the CC genotype after the exclusion diet. CT heterozygotes had intermediate concentrations. The trend toward higher tHcy in TT homozygotes persisted throughout the study but was less marked with increasing folate intake (TT compared with CC after supplementation, P = 0.097). CONCLUSIONS: A folate-rich diet including folic acid-fortified foods or low-dose supplements effectively increases folate status. TT homozygotes require higher folate intakes than do individuals with the CT or CC genotype to achieve similar tHcy concentrations but are responsive to folate intervention. PMID- 12081833 TI - Hypovitaminosis D prevalence and determinants among African American and white women of reproductive age: third National Health and Nutrition Examination Survey, 1988-1994. AB - BACKGROUND: Recent reports of rickets among African American children drew attention to the vitamin D status of these infants and their mothers. African American women are at higher risk of vitamin D deficiency than are white women, but few studies have examined determinants of hypovitaminosis D in this population. OBJECTIVE: We examined the prevalence and determinants of hypovitaminosis D among African American and white women of reproductive age. DESIGN: We examined 1546 African American women and 1426 white women aged 15-49 y who were not pregnant and who participated in the third National Health and Nutrition Examination Survey (1988-1994). Hypovitaminosis D was defined as a serum 25-hydroxyvitamin D concentration < or =37.5 nmol/L. Multiple logistic regression was used to examine the independent association of dietary, demographic, and behavioral determinants of hypovitaminosis D. RESULTS: The prevalence of hypovitaminosis D was 42.4 +/- 3.1% ( +/- SE) among African Americans and 4.2 +/- 0.7% among whites. Among African Americans, hypovitaminosis D was independently associated with consumption of milk or breakfast cereal <3 times/wk, no use of vitamin D supplements, season, urban residence, low body mass index, and no use of oral contraceptives. Even among 243 African Americans who consumed the adequate intake of vitamin D from supplements (200 IU/d), 28.2 +/- 2.7% had hypovitaminosis D. CONCLUSIONS: The high prevalence of hypovitaminosis D among African American women warrants further examination of vitamin D recommendations for these women. The determinants of hypovitaminosis D among women should be considered when these women are advised on dietary intake and supplement use. PMID- 12081834 TI - Carotenoid composition of human milk during the first month postpartum and the response to beta-carotene supplementation. AB - BACKGROUND: Information is lacking regarding normal changes in milk carotenoid concentrations in healthy, well-nourished women during the first month of lactation. OBJECTIVES: This study investigated milk carotenoid concentrations during days 4-32 postpartum and assessed the effects of maternal beta-carotene supplementation. DESIGN: Subjects (n = 21; aged 19-39 y) were randomly assigned to receive beta-carotene (30 mg/d) or placebo from days 4 to 32 postpartum. Each subject provided 8 diet records and 8 milk samples during the study. Diet records were analyzed for energy, macronutrients, vitamins A and E, and carotenoids. Milk samples were analyzed with HPLC for concentrations of carotenoids, retinol, and alpha-tocopherol. Data were analyzed by using repeated-measures analysis and orthogonal contrasts. RESULTS: No significant differences in average dietary intakes, body mass index, age, or parity were found between groups at baseline or after supplementation. Milk carotenoid concentrations decreased over time (P < 0.01), as did retinol and alpha-tocopherol concentrations (P < 0.003). Concentrations of most carotenoids decreased to those reported for mature milk by day 32 postpartum. Milk lutein concentrations remained elevated throughout the study compared with values reported for mature milk, whereas plasma lutein concentrations decreased significantly over time. beta-carotene supplementation did not significantly change the milk concentrations of beta-carotene, the other carotenoids, retinol, or alpha-tocopherol. CONCLUSIONS: The lack of increase in milk beta-carotene despite supplementation suggests that transitional milk may be already nearly saturated with beta-carotene. The elevated milk lutein concentration and simultaneous decrease in plasma lutein suggest that lutein metabolism may be altered during early lactation. PMID- 12081836 TI - Variation in newborn size according to pregnancy weight change by trimester. AB - BACKGROUND: The timing of maternal weight change in pregnancy may be an important determinant of the newborn's size. OBJECTIVE: The purpose was to identify effects of maternal weight change by trimester on newborn size, under the hypothesis that low weight gain early in pregnancy predicts proportionately smaller newborns. DESIGN: Women planning to become pregnant were followed by clinic visits and questionnaires through delivery. This study includes 389 women and their singleton infants born at a gestational age of > or = 241 d. RESULTS: In multiple regression analyses including a variety of potential confounders, maternal weight gain in the first and second trimesters predicted newborn weight (1-kg weight gain in the first trimester predicted a 31-g increase in newborn weight, P < 0.0007, and 1-kg weight gain in the second trimester predicted a 26-g increase in newborn weight, P < 0.007), but weight gain in the third trimester did not. Newborn ponderal index (in kg/m(3)) was predicted by weight gain in the first (1 kg weight gain predicted an added 0.21 units, P < 0.0003) and third (1-kg weight gain predicted an added 0.12 units, P < 0.03) trimesters but not in the second trimester. Newborn weight was 211 g lower (P < 0.006) and ponderal index 1.2 units lower (P < 0.02) in infants born to women who lost weight in the first trimester. CONCLUSIONS: The use of measured prepregnancy weight in tests of the effect on newborn size of weight gain by time in pregnancy produces different results than does the use of recalled prepregnancy weight. Maternal weight change in the first trimester of pregnancy more strongly influences newborn size than does weight change in the second or third trimester. PMID- 12081835 TI - Iron absorption in breast-fed infants: effects of age, iron status, iron supplements, and complementary foods. AB - BACKGROUND: Iron supplements are often recommended for older breast-fed infants, but little is known about factors affecting iron absorption from human milk or supplements. OBJECTIVE: We investigated the effects of age, iron status, and iron intake on iron absorption in healthy, term, breast-fed infants. DESIGN: Twenty five infants were randomly assigned to receive either 1) iron supplements (1 mg x kg(-1) x d(-1)) from 4 to 9 mo of age, 2) placebo from 4 to 6 mo and iron supplements from 6 to 9 mo, or 3) placebo from 4 to 9 mo. Infants were exclusively breast-fed to 6 mo and partially breast-fed to 9 mo of age. Iron absorption was assessed by giving (58)Fe with mother's milk at 6 and 9 mo. Blood samples were obtained at 4, 6, and 9 mo, and complementary food intake was recorded at 9 mo. RESULTS: At 6 mo, mean (+/-SD) fractional iron absorption from human milk was relatively low (16.4 +/- 11.4%), with no significant difference between iron-supplemented and unsupplemented infants. At 9 mo, iron absorption from human milk remained low in iron-supplemented infants (16.9 +/- 9.3%) but was higher (P = 0.01) in unsupplemented infants (36.7 +/- 18.9%). Unexpectedly, iron absorption at 9 mo was not correlated with iron status but was significantly correlated with intake of dietary iron, including supplemental iron. CONCLUSIONS: Changes in the regulation of iron absorption between 6 and 9 mo enhance the infant's ability to adapt to a low-iron diet and provide a mechanism by which some, but not all, infants avoid iron deficiency despite low iron intakes in late infancy. PMID- 12081837 TI - Household food security and nutritional status of Hispanic children in the fifth grade. AB - BACKGROUND: Food insecurity is a critical variable for understanding the nutritional status of low-income populations. However, limited research is available on the relation between household food insecurity and children's nutritional status. OBJECTIVE: Our objective was to examine the relations among household food insecurity, household food supplies, and school-age children's dietary intakes and body mass indexes (BMIs). DESIGN: A sample of 124 predominantly Hispanic, fifth-grade children and their mothers were surveyed as part of a school-based obesity-prevention program. Data on the children's weights and heights were collected and three 24-h dietary recalls were conducted. The mothers provided reports of household food insecurity and household food supplies. RESULTS: Food insecurity was negatively associated with the children's BMIs and household food supplies but not with the children's food intakes. However, a secondary analysis showed that as payday approached, children from the most food-insecure households had significant decreases in energy intakes and meat consumption. CONCLUSIONS: This is one of the first studies to report a significant association between food insecurity and children's nutritional status. The ages and sex-adjusted BMIs of the food-insecure children were lower than those of the food-secure children but were still within the normal range. The lower BMIs in the food-insecure children may have been due to short-term, yet periodic food restrictions that resulted as household food supplies diminished before payday. Future research is needed to assess the physiologic and psychological effects of periodic food restriction on children's health. PMID- 12081838 TI - Body composition in children with sickle cell disease. AB - BACKGROUND: Impaired growth, poor nutritional status, and delayed skeletal and sexual maturation are common in children with sickle cell disease (SCD), yet the nature of associated body-composition deficits has not been fully described. OBJECTIVE: The objective was to assess growth, nutritional status, and body composition in 36 African American children with type SS SCD (20 females and 16 males) and 30 healthy control children (15 females and 15 males) of similar age (5-18 y) and ethnicity. DESIGN: Height, weight, bone age, pubertal status, skinfold thickness, and arm circumference were assessed. Height and weight were converted to z scores by comparison with national reference data and skinfold thickness measurements were converted to z scores by comparison with African American- specific reference data. Fat-free mass (FFM) and fat mass (FM) were estimated by using 4 methods. Prepubertal children, pubertal males, and pubertal females were analyzed separately. RESULTS: Relative to the control subjects and to a national sample, children with SCD had significantly lower z scores for weight, height, arm circumference, and upper arm fat and muscle areas. Relative skeletal maturation was significantly delayed. After adjustment for age, children with SCD had significantly lower FM (prepubertal children and pubertal males only) and FFM (all 3 groups). CONCLUSIONS: Children with SCD have impaired growth, delayed puberty, and poor nutritional status. Low z scores for upper arm fat area indicate deficits in fat (energy) stores, and low FFM coupled with low upper arm muscle area indicate muscle wasting and low protein stores. These body composition abnormalities suggest that the nutritional needs of the African American children with SCD were not being met. PMID- 12081839 TI - Eating in the absence of hunger and overweight in girls from 5 to 7 y of age. AB - BACKGROUND: Eating when exposed to large portions of palatable foods in the absence of hunger has been suggested to contribute to overweight. OBJECTIVE: This research evaluated whether young girls' eating in the absence of hunger was stable across a 2-y period in middle childhood, was associated with an increased risk of overweight, and could be predicted by parents' use of restriction in child feeding. DESIGN: The participants were 192 non-Hispanic white girls and their parents, assessed when the girls were 5 and 7 y of age. The girls' eating when exposed to palatable foods in the absence of hunger was measured after they consumed a standard lunch and indicated that they were no longer hungry. RESULTS: Eating in the absence of hunger showed moderate stability across the 2-y period for most of the girls. The girls who ate large amounts of snack foods in the absence of hunger at 5 and 7 y of age were 4.6 times as likely to be overweight at both ages. Parents' reports of restricting their daughter's access to foods at age 5 y predicted girls' eating in the absence of hunger at age 7 y, even when the girls' weight status and eating in the absence of hunger at age 5 y were controlled for. CONCLUSIONS: This study provides the first evidence that young girls' eating in the absence of hunger may represent a stable phenotypic behavior of young overweight girls. In addition, these findings are consistent with previous work indicating that parents' restrictive feeding practices may contribute to this behavior. PMID- 12081840 TI - Acute ingestion of a meal rich in n-3 polyunsaturated fatty acids results in rapid gastric emptying in humans. AB - BACKGROUND: n-3 Polyunsaturated fatty acids (PUFAs) have proven benefits for both the development of atherosclerosis and inflammatory conditions. The effects on atherosclerosis may be partly mediated by the observed reduction in fasting and postprandial triacylglycerol concentrations after both acute and chronic n-3 PUFA ingestion. OBJECTIVE: The aim of this study was to assess gastric emptying and gastrointestinal hormone release after the consumption of mixed meals rich in n-3 PUFAs or other classes of fatty acids. DESIGN: Ten healthy women (aged 50-62 y) completed 4 separate study visits in a single-blind, randomized design. On each occasion, subjects consumed 40 g oil rich in either saturated fatty acids, monounsaturated fatty acids, n-6 PUFAs, or n-3 PUFAs as part of a mixed meal. [1 (13)C]Octanoic acid (100 mg) was added to each oil. Gastric emptying was assessed by a labeled octanoic acid breath test, and concentrations of gastrointestinal hormones and plasma lipids were measured. RESULTS: Recovery of (13)C in breath was enhanced after n-3 PUFA ingestion (P < 0.005). The cholecystokinin response after the n-3 PUFA meal was significantly delayed (P < 0.001), and the glucagon like peptide 1 response was significantly reduced (P < 0.05). CONCLUSION: The inclusion of n-3 PUFAs in a meal alters the gastric emptying rate, potentially as the result of changes in the pattern of cholecystokinin and glucagon-like peptide 1 release. PMID- 12081841 TI - Serum concentrations of sex hormone binding globulin are elevated in kwashiorkor and anorexia nervosa but not in marasmus. AB - BACKGROUND: Customary blood protein markers for malnutrition are of limited value in the diagnosis of protein-energy malnutrition or anorexia nervosa in children and in the follow-up to refeeding in such children. OBJECTIVES: For these diseases, we compared the diagnostic value of sex hormone binding globulin (SHBG) with that of albumin, transferrin, transthyretin, and retinal binding protein and determined the relations between concentrations of insulin, insulin-like growth factor I, and SHBG. DESIGN: SHBG was assayed in children with protein-energy malnutrition (29 children with kwashiorkor and 28 with marasmus), in 29 anorectic girls (before and after refeeding), and in age- and sex-matched control subjects. RESULTS: Mean (+/-SE) serum SHBG concentrations were higher in the children with kwashiorkor (0.18 +/- 0.07 micromol/L) than in the children with marasmus (0.11 +/- 0.05 micromol/L, P < 0.0001) or the control subjects (0.11 +/- 0.03 micromol/L, P < 0.0005). In the children with anorexia nervosa before weight gain, serum SHBG concentrations were significantly higher (0.10 +/- 0.04 micromol/L) than in the age-matched control subjects (0.06 +/- 0.03 micromol/L, P < 0.001) and decreased significantly after 30 d of refeeding (0.04 +/- 0.01 micromol/L, P < 0.0001). This decrease was negatively correlated with insulin like growth factor I but not with insulin. Mean serum SHBG concentrations were influenced neither by inflammation, as indicated when C-reactive protein was used as a marker (0.27 +/- 0.27, 0.34 +/- 0.42, and <0.04 micromol/L in the children with marasmus, kwashiorkor, and anorexia nervosa, respectively), nor by glomerular filtration, as indicated when cystatin-C was used as a marker (68.46 +/- 23.08, 66.90 +/- 43.08, and 49.23 +/- 7.69 micromol/L, respectively). CONCLUSIONS: The high SHBG concentration observed in anorexia nervosa and kwashiorkor seems to be of multifactorial origin. For these 2 diseases, SHBG is a reliable marker of nutritional status, is unrelated to either C-reactive protein or cystatin-C, and may be helpful in distinguishing kwashiorkor from marasmus and as a follow-up marker after refeeding. PMID- 12081842 TI - Bone mineral density and dietary patterns in older adults: the Framingham Osteoporosis Study. AB - BACKGROUND: Several nutrients are known to affect bone mineral density (BMD). However, these nutrients occur together in foods and dietary patterns, and the overall effects of dietary choices are not well understood. OBJECTIVE: We evaluated associations between dietary patterns and BMD in older adults. DESIGN: Of the original Framingham Heart Study subjects, 907 aged 69-93 y completed food frequency questionnaires as part of an osteoporosis study. We defined dietary patterns by cluster analysis. BMD was measured at the proximal right femur (femoral neck, trochanter, Ward's area) with a dual-photon absorptiometer and at the 33% radial shaft with a single-photon absorptiometer. We regressed BMD measures onto the cluster variable, adjusting for potential confounders. RESULTS: Six dietary patterns were identified, with relatively greater proportions of intake from meat, dairy, and bread; meat and sweet baked products; sweet baked products; alcohol; candy; and fruit, vegetables, and cereal. After adjustment for multiple comparisons, men in the last group had significantly (P = 0.05) greater BMD than did 2-4 other groups at the hip sites and the candy group at the radius. Men in the candy group had significantly (P < 0.05) lower BMD than did those in the fruit, vegetables, and cereal group for 3 of the 4 sites. Women in the candy group had significantly (P < 0.01) lower BMD than did all but one other group at the radius. CONCLUSIONS: Dietary pattern is associated with BMD. High fruit and vegetable intake appears to be protective in men. High candy consumption was associated with low BMD in both men and women. PMID- 12081843 TI - Glutamine-enriched enteral nutrition decreases infectious complications in trauma patients. PMID- 12081845 TI - Clinical nutrition: commercially or clinically essential? PMID- 12081847 TI - Dairy products, calcium, and 1,25-dihydroxyvitamin D(3) concentrations. PMID- 12081849 TI - The glycemic index at 20 y. PMID- 12081850 TI - Glycemic index: overview of implications in health and disease. AB - The glycemic index concept is an extension of the fiber hypothesis, suggesting that fiber consumption reduces the rate of nutrient influx from the gut. The glycemic index has particular relevance to those chronic Western diseases associated with central obesity and insulin resistance. Early studies showed that starchy carbohydrate foods have very different effects on postprandial blood glucose and insulin responses in healthy and diabetic subjects, depending on the rate of digestion. A range of factors associated with food consumption was later shown to alter the rate of glucose absorption and subsequent glycemia and insulinemia. At this stage, systematic documentation of the differences that exist among carbohydrate foods was considered essential. The resulting glycemic index classification of foods provided a numeric physiologic classification of relevant carbohydrate foods in the prevention and treatment of diseases such as diabetes. Since then, low-glycemic-index diets have been shown to lower urinary C peptide excretion in healthy subjects, improve glycemic control in diabetic subjects, and reduce serum lipids in hyperlipidemic subjects. Furthermore, consumption of low-glycemicindex diets has been associated with higher HDL cholesterol concentrations and, in large cohort studies, with decreased risk of developing diabetes and cardiovascular disease. Case-control studies have also shown positive associations between dietary glycemic index and the risk of colon and breast cancers. Despite inconsistencies in the data, sufficient, positive findings have emerged to suggest that the dietary glycemic index is of potential importance in the treatment and prevention of chronic diseases. PMID- 12081851 TI - Glycemic index, glycemic load, and risk of type 2 diabetes. AB - The possibility that high, long-term intake of carbohydrates that are rapidly absorbed as glucose may increase the risk of type 2 diabetes has been a long standing controversy. Two main mechanisms have been hypothesized, one mediated by increases in insulin resistance and the other by pancreatic exhaustion as a result of the increased demand for insulin. During the past decade, several lines of evidence have collectively provided strong support for a relation between such diets and diabetes incidence. In animals and in short-term human studies, a high intake of carbohydrates with a high glycemic index (a relative measure of the incremental glucose response per gram of carbohydrate) produced greater insulin resistance than did the intake of low-glycemic-index carbohydrates. In large prospective epidemiologic studies, both the glycemic index and the glycemic load (the glycemic index multiplied by the amount of carbohydrate) of the overall diet have been associated with a greater risk of type 2 diabetes in both men and women. Conversely, a higher intake of cereal fiber has been consistently associated with lower diabetes risk. In diabetic patients, evidence from medium term studies suggests that replacing high-glycemic-index carbohydrates with a low glycemic-index forms will improve glycemic control and, among persons treated with insulin, will reduce hypoglycemic episodes. These dietary changes, which can be made by replacing products made with white flour and potatoes with whole grain, minimally refined cereal products, have also been associated with a lower risk of cardiovascular disease and can be an appropriate component of recommendations for an overall healthy diet. PMID- 12081852 TI - Glycemic index and obesity. AB - Although weight loss can be achieved by any means of energy restriction, current dietary guidelines have not prevented weight regain or population-level increases in obesity and overweight. Many high-carbohydrate, low-fat diets may be counterproductive to weight control because they markedly increase postprandial hyperglycemia and hyperinsulinemia. Many high-carbohydrate foods common to Western diets produce a high glycemic response [high-glycemic-index (GI) foods], promoting postprandial carbohydrate oxidation at the expense of fat oxidation, thus altering fuel partitioning in a way that may be conducive to body fat gain. In contrast, diets based on low-fat foods that produce a low glycemic response (low-GI foods) may enhance weight control because they promote satiety, minimize postprandial insulin secretion, and maintain insulin sensitivity. This hypothesis is supported by several intervention studies in humans in which energy-restricted diets based on low-GI foods produced greater weight loss than did equivalent diets based on high-GI foods. Long-term studies in animal models have also shown that diets based on high-GI starches promote weight gain, visceral adiposity, and higher concentrations of lipogenic enzymes than do isoenergetic, macronutrientcontrolled, low-GI-starch diets. In a study of healthy pregnant women, a high-GI diet was associated with greater weight at term than was a nutrient-balanced, low-GI diet. In a study of diet and complications of type 1 diabetes, the GI of the overall diet was an independent predictor of waist circumference in men. These findings provide the scientific rationale to justify randomized, controlled, multicenter intervention studies comparing the effects of conventional and low-GI diets on weight control. PMID- 12081853 TI - Glycemic index and heart disease. AB - A diet high in carbohydrates with high glycemic indexes (GI) and glycemic load were linked to risk of coronary heart disease development in women in a large prospective study. Two cross-sectional studies showed that low-GI diets are associated with high HDL-cholesterol concentrations, especially in women. In a tightly controlled study of patients with type 2 diabetes, serum total cholesterol, LDL cholesterol, and apolipoprotein B concentrations fell more significantly after a low-GI diet than after a high-GI diet. In the same study, plasminogen activator inhibitor-1 concentrations were reduced by 58% after the low-GI diet. Insulin-stimulated glucose uptake by adipocytes was significantly higher in patients undergoing coronary artery bypass graft surgery after 4 wk of consuming a low-GI diet than after consuming a high-GI diet. The effects of low GI diets may be mediated by changes in postprandial fatty acid concentrations or by hormonal signals from adipocytes, but a possible association of low-GI diets with some other dietary factor such as chromium must not be excluded. Proof of the clinical value of low-GI diets awaits prospective trials, which should include short-term observations covering periods of metabolic stress induced by surgery as well as long-term trials with clinical endpoints. PMID- 12081854 TI - Glycemic index and disease. AB - It has been suggested that foods with a high glycemic index are detrimental to health and that healthy people should be told to avoid these foods. This paper takes the position that not enough valid scientific data are available to launch a public health campaign to disseminate such a recommendation. This paper explores the glycemic index and its validity and discusses the effect of postprandial glucose and insulin responses on food intake, obesity, type 1 diabetes, and cardiovascular disease. Presented herein are the reasons why it is premature to recommend that the general population avoid foods with a high glycemic index. PMID- 12081856 TI - Tissue engineering and reparative medicine. AB - Reparative medicine is a critical frontier in biomedical and clinical research. The National Institutes of Health Bioengineering Consortium (BECON) convened a symposium titled "Reparative Medicine: Growing Tissues and Organs," which was held on June 25 and 26, 2001 in Bethesda, Maryland. The relevant realms of cells, molecular signaling, extracellular matrix, engineering design principles, vascular assembly, bioreactors, storage and translation, and host remodeling and the immune response that are essential to tissue engineering were discussed. This overview of the scientific program summarizes the plenary talks, extended poster presentations and breakout session reports with an emphasis on scientific and technical hurdles that must be overcome to achieve the promise of restoring, replacing, or enhancing tissue and organ function that tissue engineering offers. PMID- 12081857 TI - In vitro systems for tissue engineering. AB - Tissue engineering, by necessity, encompasses a wide array of experimental directions and scientific disciplines. In vitro tissue engineering involves the manipulation of cells in vitro, prior to implantation into the in vivo environment. In contrast, in vivo tissue engineering relies on the body's natural ability to regenerate over non-cell-seeded biomaterials. Cells, biomaterials, and controlled incubation conditions all play important roles in the construction and use of modern in vitro systems for tissue engineering. Gene delivery is also an important factor for controlling or supporting the function of engineered cells both in vitro and post implantation, where appropriate. In this review, systems involved in the context of in vitro tissue engineering are addressed, including bioreactors, cell-seeded constructs, cell encapsulation, and gene delivery. Emphasis is placed upon investigations that are more directly linked to the treatment of clinical conditions. PMID- 12081858 TI - The use of cells in reparative medicine. AB - Cells are the functional elements of reparative medicine and tissue engineering. The use of living cells as a therapy presents several challenges. These include identification of a suitable source, development of adequate methods, and proof of safety and efficacy. We are now well aware that stem or pluripotent cells offer an exciting potential source for a host of functional cell types. Their true potential will only be realized through continued effort to increase basic scientific understanding at all levels, the development of adequate methods to achieve a functional phenotype, and attention to safety issues associated with adequate control of cell localization, proliferation, and differentiation. There is also new understanding regarding the immunology of parenchymal cells and new promising approaches to immune modulation, which will open the door to broader therapies using allogeneic cell sources without prohibitive immune suppression. Control of cell growth and phenotypic expression does not end in the culture vessel, but goes beyond to the patient. A living therapy is not static but dynamic, as is the host response. The cells or tissue construct in most cases will not behave as a whole-organ transplant. It is therefore important that we understand a cell or tissue therapy's ability to react and interact within the host since clinical effectiveness has proven to be one of the most difficult milestones to achieve. A living cell therapy offers great potential to alter the human condition, encompassing alteration of the current biological state of a targeted tissue or organ, augmentation of depleted or lost function, or absolute functional tissue replacement. The extent to which we are able to achieve effective cell therapies will depend on assimilating a rapidly developing base of scientific knowledge with the practical considerations of design, delivery, and host response. PMID- 12081859 TI - Reversal of established autoimmune diabetes by in situ beta-cell regeneration. PMID- 12081860 TI - Pancreatic islet cell replacement: successes and opportunities. PMID- 12081861 TI - Building animals from stem cells. PMID- 12081862 TI - Cells for repair: breakout session summary. PMID- 12081863 TI - Genetic approaches to craniofacial tissue repair. AB - This review discusses in some detail the opportunities and challenges of applying gene therapy to the important clinical problem of wound repair and regeneration. PMID- 12081864 TI - Genetic approaches to tissue repair. PMID- 12081865 TI - Gene transfer in tissue regeneration and reparative medicine. PMID- 12081866 TI - Gene-based approaches to the treatment of hemophilia. PMID- 12081868 TI - Genetic approaches to tissue repair. PMID- 12081867 TI - Genetic approaches to the repair of connective tissues. PMID- 12081869 TI - Genetic approaches to tissue engineering: breakout session summary. PMID- 12081870 TI - Different osteochondral potential of clonal cell lines derived from adult human trabecular bone. AB - Cells derived from human trabecular bones have been shown to have multipotential differentiation ability along osteogenic, chondrogenic, and adipogenic lineages. In this study, we have derived two clonal sublines of human trabecular bone cells by means of stable transduction with human papilloma virus E6/E7 genes. Our results showed that these clonal sublines differ in their osteochondral potential, but are equally adipogenic, indicative of the heterogeneous nature of the parental cell population. The availability of these cell lines should be useful for the analysis of the mechanisms regulating the differentiation of adult mesenchymal progenitor cells. PMID- 12081872 TI - Emerging design principles in biomaterials and scaffolds for tissue engineering. AB - Biomaterials and scaffolds play an essential role in tissue engineering by guiding new tissue growth in vivo and in vitro. While adaptation of existing surgical materials has fulfilled some needs in the field, new applications demand better control of bulk properties such as degradation and of surface properties that control cell interactions. Advances in molecular cell biology are driving the incorporation of new biological moieties into materials, and a set of design principles based on quantitative analysis of key cellular processes involved in regeneration is emerging. At the same time, new materials-processing methodologies are emerging to allow fabrication of these fragile materials into devices appropriate for delivery. PMID- 12081871 TI - Therapeutic potential of implanted tissue-engineered bioartificial muscles delivering recombinant proteins to the sheep heart. AB - Tissue-engineered primary adult sheep muscle cells genetically engineered to express either rhVEGF or rhIGF-1 secreted the bioactive proteins locally in the sheep heart for at least 30 days. PMID- 12081874 TI - Hybrid, composite, and complex biomaterials. PMID- 12081873 TI - Biomaterials and scaffolds in reparative medicine. AB - Most approaches currently pursued or contemplated within the framework of reparative medicine, including cell-based therapies, artificial organs, and engineered living tissues, are dependent on our ability to synthesize or otherwise generate novel materials, fabricate or assemble materials into appropriate 2-D and 3-D forms, and precisely tailor material-related physical and biological properties so as to achieve a desired clinical response. This paper summarizes the scientific and technological opportunities within the fields of biomaterials science and molecular engineering that will likely establish new enabling technologies for cellular and molecular therapies directed at the repair, replacement, or reconstruction of diseased or damaged organs and tissues. PMID- 12081875 TI - Biomaterials in reparative medicine: biorelevant structure-property analysis. PMID- 12081876 TI - Bioscaffolds for tissue repair: breakout session summary. PMID- 12081877 TI - Manufacturing and characterization of 3-d hydroxyapatite bone tissue engineering scaffolds. AB - Internal architecture has a direct impact on the mechanical and biological behaviors of porous hydroxyapatite (HA) implants. However, traditional processing methods provide very minimal control in this regard. This paper reviews a novel processing technique developed in our laboratory for fabricating scaffolds with controlled internal architectures. The preliminary mechanical property and in vivo evaluation of these scaffolds are also presented. PMID- 12081878 TI - Biological response of chondrocytes to hydrogels. AB - Primary bovine chondrocytes were encapsulated in alginate and alginate combined with cartilage matrix extract, Cartrigel, for the purpose of cartilage tissue engineering. The cell constructs were incubated in vitro and gene expression of cartilage-specific extracellular matrix molecules was quantitated and localized with in situ hybridization with a decrease in expression observed in the alginate Cartrigel constructs. Further understanding of cell response to scaffolds will allow rational design and development of hydrogels for cartilage tissue engineering. PMID- 12081879 TI - Engineering a biological joint. PMID- 12081880 TI - The efficacy of bone marrow stromal cell-seeded knitted PLGA fiber scaffold for Achilles tendon repair. PMID- 12081881 TI - Effects of alginate composition on the growth and overall metabolic activity of betaTC3 cells. PMID- 12081882 TI - Polymer/alginate amalgam for cartilage-tissue engineering. AB - Marrow stroma-derived cells (MSC) are highly proliferative, multipotential cells that have been considered as ideal candidate cells for autologous tissue engineering applications. In this study, we have characterized the chondrogenic potential of human MSCs in both a PLA/alginate amalgam and pure PLA macrostructure as model three-dimensional constructs to support both chondrogenic differentiation and proliferation following TGF-beta treatment. MSCs were seeded in experimental groups that consisted of PLA-loaded constructs and PLA/alginate amalgams with and without recombinant human TGF-beta1. Chondrogenesis of the PLA and the PLA/alginate amalgam cultures was assessed at weekly intervals by histology, immunohistochemistry, scanning electron microscopy, sulfate incorporation, and RT-PCR. Chondrogenic differentiation occurs within a polymeric macrostructure with TGF-beta1 treatment as indicated by histological, immunohistochemical, sulfate incorporation, and gene expression profiles. This macrostructure can be further encased in an alginate gel/solution to optimize cell shape and to confine growth factors and cells within the polymer construct, while the polymeric scaffold provides appropriate mechanical/tissue support. The stable three-dimensional PLA/alginate amalgam represents a novel candidate system of mesenchymal chondrogenesis, which is amendable to investigation of mechanical and biological factors that normally modulate cartilage development and formation as well as a potential tissue engineering construct for cartilage repair. PMID- 12081883 TI - Accelerated and improved osteointegration of implants biocoated with bone morphogenetic protein 2 (BMP-2). AB - A concept and methodology are presented for the direct biocoating of implantable metals like titanium and stainless steel with bone morphogenetic protein 2 (BMP 2) for future applications as cementless bone or dental prostheses. Such bioactive surfaces can influence cells and tissues by chemotactic as well as juxtacrine mechanisms. Reference is made to first experiments in sheep and rabbits in which BMP-2 coatings impressively increased the osteoinductive potential of titanium implants. PMID- 12081884 TI - Molecular signaling in bioengineered tissue microenvironments. AB - Biological tissues and organs consist of specialized living cells arrayed within a complex structural and functional framework known generally as the extracellular matrix (ECM). The great diversity observed in the morphology and composition of the ECM contributes enormously to the properties and function of each organ and tissue. For example, the ECM contributes to the rigidity and tensile strength of bone, the resilience of cartilage, the flexibility and hydrostatic strength of blood vessels, and the elasticity of skin. The ECM is also important during growth, development, and wound repair: its own dynamic composition acts as a reservoir for soluble signaling molecules and mediates signals from other sources to migrating, proliferating, and differentiating cells. Artificial three-dimensional substitutes for ECM, called tissue scaffolds, may consist of natural or synthetic polymers or a combination of both. Scaffolds have been used successfully alone and in combination with cells and soluble factors to induce tissue formation or promote tissue repair. Appropriate numbers of properly functioning living cells are central to many tissue-engineering strategies, and significant efforts have been made to identify and propagate pluripotent stem cells and lineage-restricted progenitor cells. The study of these and other living cells in artificial microenvironments, in turn, has led to the identification of signaling events important for their controlled proliferation, proper differentiation, and optimal function. PMID- 12081885 TI - Cell-cell and cell-extracellular matrix adhesion receptors. PMID- 12081886 TI - Convergence of molecular signaling and tissue engineering. PMID- 12081887 TI - The role of extracellular matrix heparan sulfate glycosaminoglycan in the activation of growth factor signaling pathways. PMID- 12081888 TI - Cell adhesion molecules activate signaling networks that influence proliferation, gene expression, and differentiation. PMID- 12081889 TI - Molecular signaling. PMID- 12081890 TI - Mechanical signaling. PMID- 12081891 TI - Molecular signaling: breakout session summary. PMID- 12081892 TI - The Provant Wound Closure System induces activation of p44/42 MAP kinase in normal cultured human fibroblasts. PMID- 12081893 TI - p38 MAP kinase regulation of AP-2 binding in TGF-beta1-stimulated chondrogenesis of human trabecular bone-derived cells. AB - Collagenase-treated, explanted human trabecular-bone chips are an excellent source of osteoblast-like cells. We have recently shown the multiple differentiation potential of these cells; in addition to osteogenesis and adipogenesis, these cells also undergo chondrogenesis when maintained as high density pellet cultures (250,000 cells/pellet) in a serum-free, chemically defined medium stimulated with TGF-beta1 (10 ng/mL). In this investigation, we have analyzed how transactivating nuclear transcription factors, specifically AP 2 and SP-1, may interact with common cis-acting elements found in the regulatory region of cartilage-specific genes as part of the signal transduction mechanism of TGF-beta1 and p38 during chondrogenesis of human trabecular bone-derived multipotential cells. Both TGF-beta1 stimulation and p38 MAP kinase activation affect the binding of AP-2 as well as SP-1 to oligonucleotides with sequence similarity to the overlapping AP-2/SP-1 sites found in the putative 52-bp immediate upstream regulatory region and the 5'-untranslated region of the human aggrecan gene. Electrophoretic mobility shift assays show that TGF-beta1 treatment of the bone-derived cells inhibits AP-2 DNA binding but enhances the DNA binding ability of SP-1. Additionally, treatment of these TGF-beta1 stimulated cells with p38 MAP kinase inhibitor, SB203580, rescued the AP-2 DNA binding but did not affect SP-1 DNA binding. These findings indicate that AP-2 DNA binding is the target of both TGF-beta1 and p38 MAP kinase signaling pathways and suggest a possible signal transduction cascade whereby TGF-beta1 induction of chondrogenesis involves the activation of p38 MAP kinase and the subsequent inhibition of DNA binding by AP-2, thereby preventing the transcriptional repression of the aggrecan gene. PMID- 12081895 TI - Tissue engineering: functional assessment and clinical outcome. AB - The issues that should be considered as part of the design and evaluation of tissue engineering constructs with respect to their targeted clinical application are reviewed. This paper provides a general framework for the process of bringing tissue-engineering constructs from the laboratory bench to the patient's bedside, rather than presenting a detailed review of the engineering or biologic principles or mechanisms that are necessary for successful tissue engineering. Many of the principles are animated by using examples from current studies being developed in my laboratory or those of my collaborators. In all likelihood, multiple solutions or approaches will be found that lead to successful tissue engineering constructs. The focus here is on the identification of critical parameters to be considered rather than specific design solutions. The review is therefore organized to reflect feasible sequences of activities formulated to take tissue engineering from concept to clinical reality. PMID- 12081894 TI - TGF-beta1-stimulated osteoblasts require intracellular calcium signaling for enhanced alpha5 integrin expression. AB - The osteoactive factor, transforming growth factor beta1 (TGF-beta1), influences osteoblast activity and bone function. We recently characterized a Smad independent TGF-beta1-induced Ca(2+) signal in human osteoblasts (HOB) and demonstrated its importance in cell adhesion. Here, we further elucidate the role of the TGF-beta1 Ca(2+) signal in the mechanics of HOB adhesion. Osteoblast interaction with fibronectin (FN) through alpha5beta1 integrin is principally responsible for osteoblast-substrate adhesion. Our results show that the TGF beta1 intracellular Ca(2+) signal is responsible, in part, for stimulation of alpha5 integrin expression, but not beta1 integrin or FN expression. Increased alpha5 integrin protein and mRNA expression was seen as early as 12 h after TGF beta1 treatment, but was inhibited by cotreatment with nifedipine, a Ca(2+) channel blocker. TGF-beta1 increased both FN and beta1 integrin protein production within 48 h, independent of nifedipine cotreatment. Immunofluorescence observations revealed that TGF-beta1 increased alpha5 integrin staining, clustering, and colocalization with the actin cytoskeleton, effects that were blocked by nifedipine. The TGF-beta1 Ca(2+) signal, a pathway crucial for HOB adhesion, enhances alpha5 integrin expression, focal contact formation, and cytoskeleton reorganization. These early events are necessary for osteoblast adhesion; thus they determine the fate of the cell and ultimately affect bone function. PMID- 12081896 TI - Functional tissue engineering: the role of biomechanics in reparative medicine. PMID- 12081897 TI - Fluorescence imaging and engineered biosensors: functional and activity-based sensing using high content screening. PMID- 12081898 TI - Functional considerations in tissue-engineering whole organs. PMID- 12081899 TI - Functional assessment and tissue design of skeletal muscle. PMID- 12081900 TI - Functional assessment of tissues with magnetic resonance imaging. PMID- 12081901 TI - Determination of responsiveness in biological systems is dependent on highly sensitive quantitative measures. PMID- 12081903 TI - Bioreactors and bioprocessing for tissue engineering. AB - Bioreactor design in tissue engineering is complex, and at the early stages of its development. Design of biologically effective, yet scalable, devices requires intimate collaboration between engineers and biologists to ensure that all aspects are considered fully. Growth conditions, harvesting time, scale-up, storage, and sterility issues all need to be considered and incorporated into the design of bioreactors. Each tissue-engineered product will likely require individualized bioreactor design. However, without a comprehensive understanding of each of these components, bioreactor design and tissue growth to manufacture product will remain at a relatively rudimentary and limited level. Increased fundamental understanding of the issues can have a dramatic impact on the ability to generate tissue-engineered product safely, economically, and in the numbers that are required to fully address the patient populations in need. PMID- 12081902 TI - Functional assessment of engineered tissues and elements of tissue design: breakout session summary. PMID- 12081904 TI - Tissue engineering as a subdivision of bioprocess engineering: reparative tissue engineering as a subspecialty of tissue engineering. PMID- 12081905 TI - Bioreactors and bioprocessing: breakout session summary. PMID- 12081906 TI - Vascular assembly in natural and engineered tissues. AB - With the advent of molecular embryology and exploitation of genetic models systems, many genes necessary for normal blood vessel formation during early development have been identified. These genes include soluble effectors and their receptors, as well as components of cell-cell junctions and mediators of cell matrix interactions. In vitro model systems (2-D and 3-D) to study paracrine and autocrine interactions of vascular cells and their progenitors have also been created. These systems are being combined to study the behavior of genetically altered cells to dissect and define the cellular role(s) of specific genes and gene families in directing the migration, proliferation, and differentiation needed for blood vessel assembly. It is clear that a complex spatial and temporal interplay of signals, including both genetic and environmental, modulates the assembly process. The development of real-time imaging and image analysis will enable us to gain further insights into this process. Collaborative efforts among vascular biologists, biomedical engineers, mathematicians, and physicists will allow us to bridge the gap between understanding vessel assembly in vivo and assembling vessels ex vivo. PMID- 12081907 TI - In vivo and in silico approaches for analysis and design of multisignal, multicomponent assembly processes in vascular systems. PMID- 12081908 TI - Vascular assembly in engineered and natural tissues. PMID- 12081909 TI - Angiogenesis: initiation and maintenance. PMID- 12081910 TI - The tissue-engineered small-diameter artery. PMID- 12081911 TI - Vascular assembly in engineered and natural tissues: breakout session summary. PMID- 12081913 TI - Neural tissue transplantation in syringomyelia: feasibility and safety. PMID- 12081912 TI - Storage and translational issues in reparative medicine. PMID- 12081915 TI - Engineering human tissues for in vivo applications. PMID- 12081914 TI - Stabilization of tissue-engineered products for transportation and extended shelf life. PMID- 12081916 TI - Cytoprotection by stabilization of cell membranes. PMID- 12081917 TI - Storage and translational issues in reparative medicine: breakout session summary. PMID- 12081918 TI - Development of artificial blood vessels: seeding and proliferation characteristics of endothelial and smooth muscle cells on biodegradable membranes. PMID- 12081919 TI - In vivo remodeling of surgically constructed vascular anastomoses: nonpenetrating, arcuate-legged clips versus standard suture. PMID- 12081921 TI - Engineering of capillary patterns in muscle by a nonmitogenic copper ribonucleoprotein angiomorphogen [angiotropin CuRNP ribokine]. PMID- 12081920 TI - Simple method for increasing cell-attachment ability of biodegradable polyester. PMID- 12081922 TI - Noninvasive monitoring of a retrievable bioartificial pancreas in vivo. PMID- 12081923 TI - In vivo studies to evaluate tissue engineering techniques. PMID- 12081925 TI - In vivo remodeling. PMID- 12081924 TI - Transition of stem cells to therapeutically functional tissue-specific cells. PMID- 12081926 TI - Regulated in vivo remodeling. PMID- 12081927 TI - Consideration of mechanical factors. PMID- 12081928 TI - Is in vivo remodeling necessary or sufficient for cellular repair of the heart? PMID- 12081929 TI - In vivo remodeling: breakout session summary. PMID- 12081930 TI - Immunological concerns with bioengineering approaches. PMID- 12081931 TI - Islet cell allotransplantation as a model system for a bioengineering approach to reparative medicine: immunological concerns. PMID- 12081932 TI - Immune responses to tissue-engineered extracellular matrix used as a bioscaffold. PMID- 12081934 TI - An innate sense of danger. PMID- 12081933 TI - Immunological barriers (opportunities?) to the use of bioengineered tissue: taking note of the innate immune system. PMID- 12081935 TI - Tolerogenic antigen-presenting cells. PMID- 12081936 TI - Chemokines in neurological trauma models. PMID- 12081937 TI - Immune response to engineered tissues and cells: breakout session summary. PMID- 12081938 TI - Tissue engineering of meniscal cartilage using perfusion culture. PMID- 12081939 TI - PROVANT Wound-Closure System accelerates closure of pressure wounds in a randomized, double-blind, placebo-controlled trial. PMID- 12081940 TI - Interconnections between inflammatory and immune responses in tissue engineering. PMID- 12081941 TI - Modulation of interferon-gamma response by dermal fibroblast extracellular matrix. PMID- 12081942 TI - Differential inflammatory and immunological responses in tissue regeneration and repair. AB - Repair and regeneration, contrasting resolutions to injury, evoke distinct inflammatory responses. Wound repair evokes a more robust peripheral leukocytosis and is accompanied by a larger inflammatory infiltrate into the wound site. Proliferation of thymocytes and splenocytes was enhanced following injury; however, the ability to be stimulated by myoblasts in coculure is lost by splenocytes harvested from rats undergoing wound repair. These data reflect differences in inflammatory and immunological responses between repair and regeneration and suggest potentially important dialogues occur between injured tissues and inflammatory cells that direct the course of injury resolution. PMID- 12081943 TI - From lab bench to market: critical issues in tissue engineering. AB - Revolutionary advances in tissue engineering are redefining approaches to tissue repair and transplantation through the creation of replacement tissues that remain biointeractive after implantation, imparting physiologic functions as well as structure to the tissue or organ damaged by disease or trauma.(1,2) Over the last decade this field has moved from "science fiction" to "science fact" with the research-oriented acceptance of its potential to regulatory approvals allowing commercial products to be available for use in many countries. The maintenance of tissue integrity, functionality, and viability from cell seeding through product manufacture, shipping, and end-use has been accomplished through innovations in design and scale-up of both tissue growth and preservation processes. These unique systems have enabled the delivery of tissue-engineered products that are uniform inter- and intra-lot, readily available as off-the shelf products, easy to use, and efficacious. Skin replacement products are the most advanced, with several tissue-engineered wound care materials on the market in the U.S. and in several international communities.(3-5) The potential impact of this field is far broader, offering novel solutions to the medical field for drug screening and development, genetic engineering, and total tissue and organ replacement. PMID- 12081944 TI - Symposium summary. PMID- 12081945 TI - Functional analysis and regulation of the divergent spuABCDEFGH-spuI operons for polyamine uptake and utilization in Pseudomonas aeruginosa PAO1. AB - A multiple-gene locus for polyamine uptake and utilization was discovered in Pseudomonas aeruginosa PAO1. This locus contained nine genes designated spuABCDEFGHI (spu for spermidine and putrescine utilization). The physiological functions of the spu genes in utilization of two polyamines (putrescine and spermidine) were analyzed by using Tn5 transposon-mediated spu knockout mutants. Growth and uptake experiments support that the spuDEFGH genes specify components of a major ABC-type transport system for spermidine uptake, and enzymatic measurements indicated that spuC encodes putrescine aminotransferase with pyruvate as the amino group receptor. Although spuA and spuB mutants showed an apparent defect in spermidine utilization, the biochemical functions of the gene products have yet to be elucidated. Assays of lacZ fusions demonstrated the presence of agmatine-, putrescine-, and spermidine-inducible promoters for the spuABCDEFGH operon and the divergently transcribed spuI gene of unknown function. Since the observed induction effect of agmatine was abolished in an aguA mutant where conversion of agmatine into putrescine was blocked, putrescine or spermidine, but not agmatine, serves as the inducer molecule of the spuA-spuI divergent promoters. S1 nuclease mappings confirmed further the induction effects of the polyamines on transcription of the divergent promoters and localized the transcription initiation sites. Gel retardation assays with extracts from the cells grown on putrescine or spermidine demonstrated the presence of a polyamine responsive regulatory protein interacting with the divergent promoter region. Finally, the absence of the putrescine-inducible spuA expression and putrescine aminotransferase (spuC) formation in the cbrB mutant indicated that the spu operons are regulated by the global CbrAB two-component system perhaps via the putative polyamine-responsive transcriptional activator. PMID- 12081946 TI - A Salmonella enterica serovar typhimurium hemA mutant is highly susceptible to oxidative DNA damage. AB - The first committed step in the biosynthesis of heme, an important cofactor of two catalases and a number of cytochromes, is catalyzed by the hemA gene product. Salmonella enterica serovar Typhimurium hemA26::Tn10d (hemA26) was identified in a genetic screen of insertion mutants that were sensitive to hydrogen peroxide. Here we show that the hemA26 mutant respires at half the rate of wild-type cells and is highly susceptible to the effects of oxygen species. Exposure of the hemA26 strain to hydrogen peroxide results in extensive DNA damage and cell death. The chelation of intracellular free iron fully abrogates the sensitivity of this mutant, indicating that the DNA damage results from the iron-catalyzed formation of hydroxyl radicals. The inactivation of heme synthesis does not change the amount of intracellular iron, but by diminishing the rate of respiration, it apparently increases the amount of reducing equivalents available to drive the Fenton reaction. We also report that hydrogen peroxide has opposite effects on the expression of hemA and hemH, the first and last genes of heme biosynthesis pathway, respectively. hemA mRNA levels decrease, while the transcription of hemH is induced by hydrogen peroxide, in an oxyR-dependent manner. The oxyR-dependent induction is suppressed under conditions that accelerate the Fenton reaction by a mechanism that is not yet understood. PMID- 12081947 TI - Inactivation of cytochrome o ubiquinol oxidase relieves catabolic repression of the Pseudomonas putida GPo1 alkane degradation pathway. AB - Expression of the alkane degradation pathway encoded by the OCT plasmid of Pseudomonas putida GPo1 is regulated by two control systems. One relies on the transcriptional regulator AlkS, which activates expression of the pathway in the presence of alkanes. The other, which is a dominant global regulation control, represses the expression of the pathway genes when a preferred carbon source is present in the growth medium in addition to alkanes. This catabolite repression control occurs through a poorly characterized mechanism that ultimately regulates transcription from the two AlkS-activated promoters of the pathway. To identify the factors involved, a screening method was developed to isolate mutants without this control. Several isolates were obtained, all of which contained mutations that mapped to genes encoding cytochrome o ubiquinol oxidase, the main terminal oxidase of the electron transport chain under highly aerobic conditions. Elimination of this terminal oxidase led to a decrease in the catabolic repression observed both in rich Luria-Bertani medium and in a defined medium containing lactate or succinate as the carbon source. This suggests that catabolic repression could monitor the physiological or metabolic status by using information from the electron transport chain or from the redox state of the cell. Since inactivation of the crc gene also reduces catabolic repression in rich medium (although not that observed in a defined medium), a strain was generated lacking both the Crc function and the cytochrome o terminal oxidase. The two mutations had an additive effect in relieving catabolic repression in rich medium. This suggests that crc and cyo belong to different regulation pathways, both contributing to catabolic repression. PMID- 12081948 TI - Family shuffling of a targeted bphA region to engineer biphenyl dioxygenase. AB - In this work we used a new strategy designed to reduce the size of the library that needs to be explored in family shuffling to evolve new biphenyl dioxygenases (BPDOs). Instead of shuffling the whole gene, we have targeted a fragment of bphA that is critical for enzyme specificity. We also describe a new protocol to screen for more potent BPDOs that is based on the detection of catechol metabolites from chlorobiphenyls. Several BphA variants with extended potency to degrade polychlorinated biphenyls (PCBs) were obtained by shuffling critical segments of bphA genes from Burkholderia sp. strain LB400, Comamonas testosteroni B-356, and Rhodococcus globerulus P6. Unlike all parents, these variants exhibited high activity toward 2,2'-, 3,3'-, and 4,4'-dichlorobiphenyls and were able to oxygenate the very persistent 2,6-dichlorobiphenyl. The data showed that the replacement of a short segment (335TFNNIRI341) of LB400 BphA by the corresponding segment (333GINTIRT339) of B-356 BphA or P6 BphA contributes to relax the enzyme toward PCB substrates. PMID- 12081949 TI - Evidence from terminal recombination gradients that FtsK uses replichore polarity to control chromosome terminus positioning at division in Escherichia coli. AB - Chromosome dimers in Escherichia coli are resolved at the dif locus by two recombinases, XerC and XerD, and the septum-anchored FtsK protein. Chromosome dimer resolution (CDR) is subject to strong spatiotemporal control: it takes place at the time of cell division, and it requires the dif resolution site to be located at the junction between the two polarized chromosome arms or replichores. Failure of CDR results in trapping of DNA by the septum and RecABCD recombination (terminal recombination). We had proposed that dif sites of a dimer are first moved to the septum by mechanisms based on local polarity and that normally CDR then occurs as the septum closes. To determine whether FtsK plays a role in the mobilization process, as well as in the recombination reaction, we characterized terminal recombination in an ftsK mutant. The frequency of recombination at various points in the terminus region of the chromosome was measured and compared with the recombination frequency on a xerC mutant chromosome with respect to intensity, the region affected, and response to polarity distortion. The use of a prophage excision assay, which allows variation of the site of recombination and interference with local polarity, allowed us to find that cooperating FtsK dependent and -independent processes localize dif at the septum and that DNA mobilization by FtsK is oriented by the polarity probably due to skewed sequence motifs of the mobilized material. PMID- 12081950 TI - The RNA polymerase alpha subunit from Sinorhizobium meliloti can assemble with RNA polymerase subunits from Escherichia coli and function in basal and activated transcription both in vivo and in vitro. AB - Sinorhizobium meliloti, a gram-negative soil bacterium, forms a nitrogen-fixing symbiotic relationship with members of the legume family. To facilitate our studies of transcription in S. meliloti, we cloned and characterized the gene for the alpha subunit of RNA polymerase (RNAP). S. meliloti rpoA encodes a 336-amino acid, 37-kDa protein. Sequence analysis of the region surrounding rpoA identified six open reading frames that are found in the conserved gene order secY (SecY) adk (Adk)-rpsM (S13)-rpsK (S11)-rpoA (alpha)-rplQ (L17) found in the alpha proteobacteria. In vivo, S. meliloti rpoA expressed in Escherichia coli complemented a temperature sensitive mutation in E. coli rpoA, demonstrating that S. meliloti alpha supports RNAP assembly, sequence-specific DNA binding, and interaction with transcriptional activators in the context of E. coli. In vitro, we reconstituted RNAP holoenzyme from S. meliloti alpha and E. coli beta, beta', and sigma subunits. Similar to E. coli RNAP, the hybrid RNAP supported transcription from an E. coli core promoter and responded to both upstream (UP) element- and Fis-dependent transcription activation. We obtained similar results using purified RNAP from S. meliloti. Our results demonstrate that S. meliloti alpha functions are conserved in heterologous host E. coli even though the two alpha subunits are only 51% identical. The ability to utilize E. coli as a heterologous system in which to study the regulation of S. meliloti genes could provide an important tool for our understanding and manipulation of these processes. PMID- 12081951 TI - Natural resistance to inhibitors of the ubiquinol cytochrome c oxidoreductase of Rubrivivax gelatinosus: sequence and functional analysis of the cytochrome bc(1) complex. AB - Biochemical analyses of Rubrivivax gelatinosus membranes have revealed that the cytochrome bc(1) complex is highly resistant to classical inhibitors including myxothiazol, stigmatellin, and antimycin. This is the first report of a strain exhibiting resistance to inhibitors of both catalytic Q(0) and Q(i) sites. Because the resistance to cytochrome bc(1) inhibitors is primarily related to the cytochrome b primary structure, the petABC operon encoding the subunits of the cytochrome bc(1) complex of Rubrivivax gelatinosus was sequenced. In addition to homologies to the corresponding proteins from other organisms, the deduced amino acid sequence of the cytochrome b polypeptide shows (i) an E303V substitution in the highly conserved PEWY loop involved in quinol/stigmatellin binding, (ii) other substitutions that could be involved in resistance to cytochrome bc(1) inhibitors, and (iii) 14 residues instead of 13 between the histidines in helix IV that likely serve as the second axial ligand to the b(H) and b(L) hemes, respectively. These characteristics imply different functional properties of the cytochrome bc(1) complex of this bacterium. The consequences of these structural features for the resistance to inhibitors and for the properties of R. gelatinosus cytochrome bc(1) are discussed with reference to the structure and function of the cytochrome bc(1) complexes from other organisms. PMID- 12081952 TI - Analysis of DNA regulatory elements required for expression of the Legionella pneumophila icm and dot virulence genes. AB - To investigate the regulation of the Legionella pneumophila icm and dot genes required for intracellular growth, a series of nine icm::lacZ fusions were constructed. These icm::lacZ fusions were found to have different levels of expression in L. pneumophila, and five of them were more highly expressed at stationary phase than at exponential phase. When the expression of these fusions in Escherichia coli was tested, all of them were found to be expressed but three of them had dramatic changes in their levels of expression in comparison to those in L. pneumophila. Site-directed and PCR random mutagenesis with these icm::lacZ fusions was used to identify DNA regulatory elements of icm genes. Four icm genes (icmT, icmP, icmQ, and icmM) that had low levels of expression in L. pneumophila were found to contain a 6-bp sequence (TATACT) essential for their expression. This sequence was shown by primer extension to serve as their -10 promoter elements. A similar sequence, which constitutes the -10 promoter elements of the icmV, icmW, and icmR genes which had high levels of expression in L. pneumophila, was also identified. In addition, regulatory elements that probably serve as binding sites for transcription regulators were found in these genes. Altogether, 12 regulatory elements, 7 of which constitute the -10 promoter elements of the icm genes, were found. Even though all the icm and dot genes are part of one system required for L. pneumophila intracellular growth and even though their promoters are probably recognized by the vegetative sigma factor, it seems that they are subjected to different regulation mediated by several regulatory factors. PMID- 12081953 TI - DNA polymerases of low-GC gram-positive eubacteria: identification of the replication-specific enzyme encoded by dnaE. AB - dnaE, the gene encoding one of the two replication-specific DNA polymerases (Pols) of low-GC-content gram-positive bacteria (E. Dervyn et al., Science 294:1716-1719, 2001; R. Inoue et al., Mol. Genet. Genomics 266:564-571, 2001), was cloned from Bacillus subtilis, a model low-GC gram-positive organism. The gene was overexpressed in Escherichia coli. The purified recombinant product displayed inhibitor responses and physical, catalytic, and antigenic properties indistinguishable from those of the low-GC gram-positive-organism-specific enzyme previously named DNA Pol II after the polB-encoded DNA Pol II of E. coli. Whereas a polB-like gene is absent from low-GC gram-positive genomes and whereas the low GC gram-positive DNA Pol II strongly conserves a dnaE-like, Pol III primary structure, it is proposed that it be renamed DNA polymerase III E (Pol III E) to accurately reflect its replicative function and its origin from dnaE. It is also proposed that DNA Pol III, the other replication-specific Pol of low-GC gram positive organisms, be renamed DNA polymerase III C (Pol III C) to denote its origin from polC. By this revised nomenclature, the DNA Pols that are expressed constitutively in low-GC gram-positive bacteria would include DNA Pol I, the dispensable repair enzyme encoded by polA, and the two essential, replication specific enzymes Pol III C and Pol III E, encoded, respectively, by polC and dnaE. PMID- 12081954 TI - Characterization of genes involved in modulation of conjugal transfer of the Bacteroides conjugative transposon CTnDOT. AB - In previous studies we identified an 18-kb region of the Bacteroides conjugative transposon CTnDOT that was sufficient for mobilization of coresident plasmids and unlinked integrated elements, as well as self-transfer from Bacteroides to Escherichia coli. When this 18-kb region was cloned on a plasmid (pLYL72), the plasmid transferred itself constitutively in the absence of a coresident conjugative transposon. However, when this plasmid was present in a Bacteroides strain containing a coresident conjugative transposon, conjugal transfer was repressed in the absence of tetracycline and enhanced in the presence of tetracycline. These results suggested that a negative and a positive regulator of conjugal transfer were encoded outside the transfer region of the CTnDOT element. In this work, a minimal and inducible transfer system was constructed and used in transfer and Western blot analyses to identify the differentially regulated genes from CTnDOT responsible for the enhancement and repression of pLYL72 conjugal transfer. Both of these regulatory functions have been localized to a region of the CTnDOT element that is essential for CTn excision. In the presence of tetracycline, the regulatory protein RteC activates the expression of a putative topoisomerase gene, exc, which in turn results in an increase in transfer protein expression and a concomitant 100- to 1,000-fold increase in the frequency of pLYL72 transfer. Our results also suggest that since exc alone cannot result in enhancement of transfer, other factors encoded upstream of exc are also required. Conversely, in the absence of tetracycline, a gene located near the 3' end of exc is responsible for the repression of transfer protein expression and also results in a 100- to 1,000-fold decrease in the frequency of pLYL72 transfer. PMID- 12081955 TI - Mutations in the CCGTTCACA DnaA box of Mycobacterium tuberculosis oriC that abolish replication of oriC plasmids are tolerated on the chromosome. AB - The origin of replication (oriC) region in some clinical strains of Mycobacterium tuberculosis is a hot spot for IS6110 elements. To understand how clinical strains with insertions in oriC can replicate their DNA, we characterized the oriC regions of some clinical strains. Using a plasmid-based oriC-dependent replication assay, we showed that IS6110 insertions that disrupted the DnaA box sequence CCGTTCACA abolished oriC activity in M. tuberculosis. Furthermore, by using a surface plasmon resonance technique we showed that purified M. tuberculosis DnaA protein binds native but not mutant DnaA box sequence, suggesting that stable interactions of the DnaA protein with the CCGTTCACA DnaA box are crucial for replication of oriC plasmids in vivo. Replacement by homologous recombination of the CCGTTCACA DnaA box sequence of the laboratory strain M. tuberculosis H37Ra with a mutant sequence did not result in nonviability. Together, these results suggest that M. tuberculosis strains have evolved mechanisms to tolerate mutations in the oriC region and that functional requirements for M. tuberculosis oriC replication are different for chromosomes and plasmids. PMID- 12081956 TI - FtsA mutants of Bacillus subtilis impaired in sporulation. AB - Spore formation in Bacillus subtilis involves a switch in the site of cell division from the midcell to a polar position. Both medial division and polar division are mediated in part by the actin-like, cytokinetic protein FtsA. We report the isolation of an FtsA mutant (FtsA(D265G)) that is defective in sporulation but is apparently unimpaired in vegetative growth. Sporulating cells of the mutant reach the stage of asymmetric division but are partially blocked in the subsequent morphological process of engulfment. As judged by fluorescence microscopy and electron microscopy, the FtsA(D265G) mutant produces normal looking medial septa but immature (abnormally thin) polar septa. The mutant was unimpaired in transcription under the control of Spo0A, the master regulator for entry into sporulation, but was defective in transcription under the control of sigmaF, a regulatory protein whose activation is known to depend on polar division. An amino acid substitution at a residue (Y264) adjacent to D265 also caused a defect in sporulation. D265 and Y264 are conserved among endospore forming bacteria, raising the possibility that these residues are involved in a sporulation-specific protein interaction that facilitates maturation of the sporulation septum and the activation of sigmaF. PMID- 12081957 TI - Cleavage of Treponema denticola PrcA polypeptide to yield protease complex associated proteins Prca1 and Prca2 is dependent on PrtP. AB - Analysis of potential virulence factors of oral spirochetes focuses on surface and secreted proteins. The Treponema denticola chymotrypsin-like protease (CTLP) is implicated in degradation of host cell molecules and contributes to tissue invasion. The CTLP complex, composed of the 72-kDa PrtP protein and two auxiliary proteins with molecular masses of approximately 40 and 30 kDa, is also involved in localization and oligomerization of the T. denticola major surface protein (Msp). The larger auxiliary protein was reported to be encoded by an open reading frame (ORF2) directly upstream of prtP. The deduced 39-kDa translation product of ORF2 contains a sequence matching the N-terminal sequence determined from one of the CTLP complex proteins. No proteins with significant homology are known, nor was information available on the third protein of the complex. DNA sequence analysis showed that ORF2 extended an additional 852 bp upstream of the reported sequence. The complete gene, designated prcA, encodes a predicted N-terminally acylated polypeptide of approximately 70 kDa. Isogenic mutants with mutations in prtP, prcA, and prcA-prtP all lacked CTLP protease activity. The prcA mutant lacked all three CTLP proteins. The prcA-prtP mutant produced only a C-terminally truncated 62-kDa PrcA protein. The prtP mutant produced a full-length 70-kDa PrcA. Immunoblot analysis of recombinant PrcA constructs confirmed that PrcA is cleaved to yield the two smaller proteins of the CTLP complex, designated PrcA1 and PrcA2. These data indicate that PrtP is required for cleavage of PrcA and suggest that this cleavage may be required for formation or stability of outer membrane complexes. PMID- 12081958 TI - The PhlA hemolysin from the entomopathogenic bacterium Photorhabdus luminescens belongs to the two-partner secretion family of hemolysins. AB - Photorhabdus is an entomopathogenic bacterium symbiotically associated with nematodes of the family Heterorhabditidae. Bacterial hemolysins found in numerous pathogenic bacteria are often virulence factors. We describe here the nucleotide sequence and the molecular characterization of the Photorhabdus luminescens phlBA operon, a locus encoding a hemolysin which shows similarities to the Serratia type of hemolysins. It belongs to the two-partner secretion (TPS) family of proteins. In low-iron conditions, a transcriptional induction of the phlBA operon was observed by using the chloramphenicol acetyltransferase reporter gene, causing an increase in PhlA hemolytic activity compared to iron-rich media. A spontaneous phase variant of P. luminescens was deregulated in phlBA transcription. The phlA mutant constructed by allelic exchange remained highly pathogenic after injection in the lepidopteran Spodoptera littoralis, indicating that PhlA hemolysin is not a major virulence determinant. Using the gene encoding green fluorescent protein as a reporter, phlBA transcription was observed in hemolymph before insect death. We therefore discuss the possible role of PhlA hemolytic activity in the bacterium-nematode-insect interactions. PMID- 12081959 TI - Network of hydrogenase maturation in Escherichia coli: role of accessory proteins HypA and HybF. AB - We have studied the roles of the auxiliary protein HypA and of its homolog HybF in hydrogenase maturation. A mutation in hypA leads to the nearly complete blockade of maturation solely of hydrogenase 3 whereas a lesion in hybF drastically but not totally reduces maturation and activity of isoenzymes 1 and 2. The residual level of matured enzymes in the hybF mutant was shown to be due to the function of HypA; HybF, conversely, was responsible for a minimal residual activity of hydrogenase 3 in the mutant hypA strain. Accordingly, a hypA DeltahybF double mutant was completely blocked in the maturation process. However, the inclusion of high nickel concentrations in the medium could restore limited activity of all three hydrogenases. The results of this study and of previous work (M. Blokesch, A. Magalon, and A. Bock, J. Bacteriol. 189:2817-2822, 2001) show that the maturation of the three functional hydrogenases from Escherichia coli is intimately connected via the activity of proteins HypA and HypC and of their homologs HybF and HybG, respectively. The results also support the suggestion of Olson et al. (J. W. Olson, N. S. Mehta, and R. J. Maier, Mol. Microbiol. 39:176-182, 2001) that HypA cooperates with HypB in the insertion of nickel into the precursor of the large hydrogenase subunit. Whereas HypA is predominantly involved in the maturation of hydrogenase 3, HybF takes over its function in the maturation of isoenzymes 1 and 2. PMID- 12081960 TI - Patterns of sequence conservation in the S-Layer proteins and related sequences in Clostridium difficile. AB - Clostridium difficile is the etiological agent of antibiotic-associated diarrhea. Among the factors that may play a role in infection are S-layer proteins (SLPs). Previous work has shown these to consist mainly of two components, resulting from the cleavage of a precursor encoded by the slpA gene. The high-molecular-weight (MW) subunit is related both to amidases from B. subtilis and to at least another 28 gene products in C. difficile strain 630. To gain insight into the functions of the SLPs and related proteins, we have further investigated the pattern of variability both at the slpA locus and at six nearby paralogs. Sequencing of the slpA gene from an S-layer group II strain and a variant S-layer group strain confirms a high degree of divergence in the low-MW SLP, which may result from diversifying selection. A highly conserved motif, however, is found at the C terminus in all low-MW subunits and may be essential for SlpA precursor cleavage. In strain 167, a variant cleavage product is present, suggesting a secondary processing site. Southern blotting analysis shows slpA-like open reading frames (ORFs) 2 to 7 to be conserved in all nine strains tested, with one exception: ORF2, which encodes a 66-kDa polypeptide coextracted at low pH with the main SLPs in strain 630, may be partially deleted in strain 167. Polymorphism within the slpA-ORF7 cluster may be more pronounced in the region proximal to the slpA gene. Unexpectedly, a high-MW subunit probe cross hybridizes to sequences outside the slpA locus, which appear to vary in number in different strains. PMID- 12081961 TI - Identification of a new class of cytochrome P450 from a Rhodococcus sp. AB - A degenerate set of PCR primers were used to clone a gene encoding a cytochrome P450 (the P450RhF gene) from Rhodococcus sp. strain NCIMB 9784 which is of unique primary structural organization. Surprisingly, analysis of the translation product revealed that the P450 is fused to a reductase domain at the C terminus which displays sequence conservation for dioxygenase reductase proteins. The reductase partner comprises flavin mononucleotide- and NADH-binding motifs and a [2Fe2S] ferredoxin-like center. The gene was engineered for heterologous expression in Escherichia coli, and conditions were found in which the enzyme was produced in a soluble form. A recombinant strain of E. coli was able to mediate the O dealkylation of 7-ethoxycoumarin in good yield, despite the absence of any recombinant redox proteins. This unprecedented finding leads us to propose that P450RhF represents the first example of a new class of cytochromes P450 in which the reducing equivalents are supplied by a novel reductase in a fused arrangement. PMID- 12081962 TI - The glycolytic flux in Escherichia coli is controlled by the demand for ATP. AB - The nature of the control of glycolytic flux is one of the central, as-yet uncharacterized issues in cellular metabolism. We developed a molecular genetic tool that specifically induces ATP hydrolysis in living cells without interfering with other aspects of metabolism. Genes encoding the F(1) part of the membrane bound (F(1)F(0)) H(+)-ATP synthase were expressed in steadily growing Escherichia coli cells, which lowered the intracellular [ATP]/[ADP] ratio. This resulted in a strong stimulation of the specific glycolytic flux concomitant with a smaller decrease in the growth rate of the cells. By optimizing additional ATP hydrolysis, we increased the flux through glycolysis to 1.7 times that of the wild-type flux. The results demonstrate why attempts in the past to increase the glycolytic flux through overexpression of glycolytic enzymes have been unsuccessful: the majority of flux control (>75%) resides not inside but outside the pathway, i.e., with the enzymes that hydrolyze ATP. These data further allowed us to answer the question of whether catabolic or anabolic reactions control the growth of E. coli. We show that the majority of the control of growth rate resides in the anabolic reactions, i.e., the cells are mostly "carbon" limited. Ways to increase the efficiency and productivity of industrial fermentation processes are discussed. PMID- 12081963 TI - Group I self-splicing intron in the recA gene of Bacillus anthracis. AB - Self-splicing introns are rarely found in bacteria and bacteriophages. They are classified into group I and II according to their structural features and splicing mechanisms. While the group I introns are occasionally found in protein coding regions of phage genomes and in several tRNA genes of cyanobacteria and proteobacteria, they had not been found in protein-coding regions of bacterial genomes. Here we report a group I intron in the recA gene of Bacillus anthracis which was initially found by DNA sequencing as an intervening sequence (IVS). By using reverse transcriptase PCR, the IVS was shown to be removable from the recA precursor mRNA for RecA that was being translated in E. coli. The splicing was visualized in vitro with labeled free GTP, indicating that it is a group I intron, which is also implied by its predicted secondary structure. The RecA protein of B. anthracis expressed in E. coli was functional in its ability to complement a recA defect. When recA-negative E. coli cells were irradiated with UV, the Bacillus RecA reduced the UV susceptibility of the recA mutant, regardless of the presence of intron. PMID- 12081964 TI - RelA protein is involved in induction of genetic competence in certain Bacillus subtilis strains by moderating the level of intracellular GTP. AB - We found that the ability to develop genetic competence of a certain relaxed (relA) aspartate-auxotrophic strain of Bacillus subtilis is significantly lower than that of the isogenic stringent (relA+) strain. Transcriptional fusion analysis utilizing a lacZ reporter gene indicated that the amount of the ComK protein, known as the key protein for competence development, is greatly reduced in the relaxed strain than in the stringent strain. We also found that the addition of decoyinine, a GMP synthetase inhibitor, induces expression of a competence gene (comG) in the relaxed strain, accompanied by a pronounced decrease in the level of intracellular GTP as measured by high-performance liquid chromatography. The transformation efficiency of the relaxed strain increased 100 fold when decoyinine was added at t0 (the transition point between exponential to stationary growth phase). Conversely, supplementation of guanosine together with decoyinine completely abolished the observed effect of adding decoyinine on competence development. Furthermore, the impaired ability of the relaxed strain for competence development was completely restored by disrupting the codY gene, which is known to negatively control comK expression. Our results indicate that the RelA protein plays an essential role in the induction of competence development at least under certain physiological conditions by reducing the level of intracellular GTP and overcoming CodY-mediated regulation. PMID- 12081966 TI - An AraC/XylS family member at a high level in a hierarchy of regulators for phenol-metabolizing enzymes in Comamonas testosteroni R5. AB - Comamonas testosteroni strain R5 expresses a higher level of phenol-oxygenating activity than any other bacterial strain so far characterized. The expression of the operon encoding multicomponent phenol hydroxylase (mPH), which is responsible for the phenol-oxygenating activity, is controlled by two transcriptional regulators, PhcS and PhcR, in strain R5. In this study, we identified a third transcriptional regulator for the mPH operon (PhcT) that belongs to the AraC/XylS family. While the disruption of phcT in strain R5 significantly reduced the expression of the mPH operon, it did not eliminate the expression. However, the disruption of phcT in strain R5 increased the expression of phcR. The phenol oxygenating activity was abolished by the disruption of phcR, indicating that PhcT alone was not sufficient to activate the expression of the mPH operon. The disruption of phcS has been shown in our previous study to confer the ability of strain R5 to express the mPH operon in the absence of the genuine substrate for mPH. PhcT was not involved in the gratuitous expression. Strain R5 thus possesses a more elaborate mechanism for regulating the mPH operon expression than has been found in other bacteria. PMID- 12081965 TI - Novel DNA-binding proteins in the cyanobacterium Anabaena sp. strain PCC 7120. AB - As an approach towards elucidation of the biochemical regulation of the progression of heterocyst differentiation in Anabaena sp. strain PCC 7120, we have identified proteins that bind to a 150-bp sequence upstream from hepC, a gene that plays a role in the synthesis of heterocyst envelope polysaccharide. Such proteins were purified in four steps from extracts of vegetative cells of Anabaena sp. Two of these proteins (Abp1 and Abp2) are encoded by neighboring genes in the Anabaena sp. chromosome. The genes that encode the third (Abp3) and fourth (Abp4) proteins are situated at two other loci in that chromosome. Insertional mutagenesis of abp2 and abp3 blocked expression of hepC and hepA and prevented heterocyst maturation and aerobic fixation of N(2). PMID- 12081967 TI - Export of L-isoleucine from Corynebacterium glutamicum: a two-gene-encoded member of a new translocator family. AB - Bacteria possess amino acid export systems, and Corynebacterium glutamicum excretes L-isoleucine in a process dependent on the proton motive force. In order to identify the system responsible for L-isoleucine export, we have used transposon mutagenesis to isolate mutants of C. glutamicum sensitive to the peptide isoleucyl-isoleucine. In one such mutant, strong peptide sensitivity resulted from insertion into a gene designated brnF encoding a hydrophobic protein predicted to possess seven transmembrane spanning helices. brnE is located downstream of brnF and encodes a second hydrophobic protein with four putative membrane-spanning helices. A mutant deleted of both genes no longer exports L-isoleucine, whereas an overexpressing strain exports this amino acid at an increased rate. BrnF and BrnE together are also required for the export of L leucine and L-valine. BrnFE is thus a two-component export permease specific for aliphatic hydrophobic amino acids. Upstream of brnFE and transcribed divergently is an Lrp-like regulatory gene required for active export. Searches for homologues of BrnFE show that this type of exporter is widespread in prokaryotes but lacking in eukaryotes and that both gene products which together comprise the members of a novel family, the LIV-E family, generally map together within a single operon. Comparisons of the BrnF and BrnE phylogenetic trees show that gene duplication events in the early bacterial lineage gave rise to multiple paralogues that have been retained in alpha-proteobacteria but not in other prokaryotes analyzed. PMID- 12081968 TI - The bacteriophage T4 transcription activator MotA interacts with the far-C terminal region of the sigma70 subunit of Escherichia coli RNA polymerase. AB - Transcription from bacteriophage T4 middle promoters uses Escherichia coli RNA polymerase together with the T4 transcriptional activator MotA and the T4 coactivator AsiA. AsiA binds tightly within the C-terminal portion of the sigma70 subunit of RNA polymerase, while MotA binds to the 9-bp MotA box motif, which is centered at -30, and also interacts with sigma70. We show here that the N terminal half of MotA (MotA(NTD)), which is thought to include the activation domain, interacts with the C-terminal region of sigma70 in an E. coli two-hybrid assay. Replacement of the C-terminal 17 residues of sigma70 with comparable sigma38 residues abolishes the interaction with MotA(NTD) in this assay, as does the introduction of the amino acid substitution R608C. Furthermore, in vitro transcription experiments indicate that a polymerase reconstituted with a sigma70 that lacks C-terminal amino acids 604 to 613 or 608 to 613 is defective for MotA dependent activation. We also show that a proteolyzed fragment of MotA that contains the C-terminal half (MotA(CTD)) binds DNA with a K(D(app)) that is similar to that of full-length MotA. Our results support a model for MotA dependent activation in which protein-protein contact between DNA-bound MotA and the far-C-terminal region of sigma70 helps to substitute functionally for an interaction between sigma70 and a promoter -35 element. PMID- 12081970 TI - Properties of 2-oxoglutarate:ferredoxin oxidoreductase from Thauera aromatica and its role in enzymatic reduction of the aromatic ring. AB - Benzoyl coenzyme A (benzoyl-CoA) reductase is a key enzyme in the anaerobic metabolism of aromatic compounds catalyzing the ATP-driven reductive dearomatization of benzoyl-CoA. The enzyme from Thauera aromatica uses a reduced 2[4Fe-4S] ferredoxin as electron donor. In this work, we identified 2 oxoglutarate:ferredoxin oxidoreductase (KGOR) as the ferredoxin reducing enzyme. KGOR activity was increased 10- to 50-fold in T. aromatica cells grown under denitrifying conditions on an aromatic substrate compared to that of cells grown on nonaromatic substrates. The enzyme was purified from soluble extracts by a 60 fold enrichment with a specific activity of 4.8 micromol min(-1) mg(-1). The native enzyme had a molecular mass of 200 +/- 20 kDa (mean +/- standard deviation) and consisted of two subunits with molecular masses of 66 and 34 kDa, suggesting an (alphabeta)(2) composition. The UV/visible spectrum was characteristic for an iron-sulfur protein; the enzyme contained 8.3 +/- 0.5 mol of Fe, 7.2 +/- 0.5 mol of acid-labile sulfur, and 1.6 +/- 0.2 mol of thiamine diphosphate (TPP) per mol of protein. The high specificity for 2-oxoglutarate and the low K(m) for ferredoxin ( approximately 10 microM) indicated that both are the in vivo substrates of the enzyme. KGOR catalyzed the isotope exchange between (14)CO(2) and C(1) of 2-oxoglutarate, representing a typical reversible partial reaction of 2-oxoacid oxidoreductases. The two genes coding for the two subunits of KGOR were found adjacent to the gene cluster coding for enzymes and ferredoxin of the catabolic benzoyl-CoA pathway. Sequence comparisons with other 2-oxoacid oxidoreductases indicated that KGOR from T. aromatica belongs to the Halobacterium type of 2-oxoacid oxidoreductases, which lack a ferredoxin-like module which contains two additional [4Fe-4S](1+/2+) clusters/monomer. Using purified KGOR, ferredoxin, and benzoyl-CoA reductase, the 2-oxoglutarate-driven reduction of benzoyl-CoA was shown in vitro. This demonstrates that ferredoxin acts as an electron shuttle between the citric acid cycle and benzoyl-CoA reductase by coupling the oxidation of the end product of the benzoyl-CoA pathway, acetyl-CoA, to the reduction of the aromatic ring. PMID- 12081969 TI - The gonococcal fur regulon: identification of additional genes involved in major catabolic, recombination, and secretory pathways. AB - In this study, we have characterized the in vitro binding of Neisseria gonorrhoeae Fur to several well-defined iron transport genes, as well as to additional genes involved in major catabolic, secretory, and recombination pathways of gonococci. The gonococcal Fur protein was recombinantly expressed in Escherichia coli HBMV119. Fur was isolated from inclusion bodies and partially purified by ion-exchange chromatography. Gonococcal Fur was found to bind to the promoter/operator region of a gene encoding the previously identified Fur regulated periplasmic binding protein (FbpA) in a metal ion-dependent fashion, demonstrating that purified Fur is functional. In silico analysis of the partially completed gonococcal genome (FA1090) identified Fur boxes in the promoters of several genes, including tonB, fur, recN, secY, sodB, hemO, hmbR, fumC, a hypothetical gene (Fe-S homolog), and the opa family of genes. By using purified gonococcal Fur, we demonstrate binding to the operator regions of tonB, fur, recN, secY, sodB, hemO, hmbR, fumC, the Fe-S homolog gene, and the opa gene family as determined by an electrophoretic mobility shift assay. While gonococcal Fur was demonstrated to bind to the promoter regions of all 11 opa genes (opaA through -K), we did not detect binding of purified E. coli Fur with 8 of the 11 opa members, indicating that target DNA sequence specificities between these two closely related proteins exist. Furthermore, we observed differences in the relative strengths of binding of gonococcal Fur for these different genes, which most likely reflect a difference in affinity between gonococcal Fur and its DNA targets. This is the first report that definitively demonstrates the binding of gonococcal Fur to its own promoter/operator region, as well as to the opa family of genes that encode surface proteins. Our results demonstrate that the gonococcal Fur protein binds to the regulatory regions of a broad array of genes and indicates that the gonococcal Fur regulon is larger than originally proposed. PMID- 12081971 TI - Activation of antibiotic biosynthesis by specified mutations in the rpoB gene (encoding the RNA polymerase beta subunit) of Streptomyces lividans. AB - We found that the biosynthesis of actinorhodin (Act), undecylprodigiosin (Red), and calcium-dependent antibiotic (CDA) are dramatically activated by introducing certain mutations into the rpoB gene that confer resistance to rifampin to Streptomyces lividans 66, which produces less or no antibiotics under normal growth conditions. Activation of Act and/or Red biosynthesis by inducing mutations in the rpoB gene was shown to be dependent on the mutation's position and the amino acid species substituted in the beta-subunit of the RNA polymerase. Mutation analysis identified 15 different kinds of point mutations, which are located in region I, II, or III of the rpoB gene and, in addition, two novel mutations (deletion of nucleotides 1287 to 1289 and a double substitution at nucleotides 1309 and 1310) were also found. Western blot analyses and S1 mapping analyses demonstrated that the expression of actII-ORF4 and redD, which are pathway-specific regulatory genes for Act and Red, respectively, was activated in the mutants able to produce Act and Red. The ActIV-ORF1 protein (an enzyme for Act biosynthesis) and the RedD protein were produced just after the upregulation of ActII-ORF4 and RedZ, respectively. These results indicate that the mutation in the rpoB gene of S. lividans, resulting in the activation of Act and/or Red biosynthesis, functions at the transcription level by activating directly or indirectly the key regulatory genes, actII-ORF4 and redD. We propose that the mutated RNA polymerase may function by mimicking the ppGpp-bound form in activating the onset of secondary metabolism in STREPTOMYCES: PMID- 12081972 TI - A repressor protein, PhaR, regulates polyhydroxyalkanoate (PHA) synthesis via its direct interaction with PHA. AB - Phasins (PhaP) are predominantly polyhydroxyalkanoate (PHA) granule-associated proteins that positively affect PHA synthesis. Recently, we reported that the phaR gene, which is located downstream of phaP in Paracoccus denitrificans, codes for a negative regulator involved in PhaP expression. In this study, DNase I footprinting revealed that PhaR specifically binds to two regions located upstream of phaP and phaR, suggesting that PhaR plays a role in the regulation of phaP expression as well as autoregulation. Many TGC-rich sequences were found in upstream elements recognized by PhaR. PhaR in the crude lysate of recombinant Escherichia coli was able to rebind specifically to poly[(R)-3-hydroxybutyrate] [P(3HB)] granules. Furthermore, artificial P(3HB) granules and 3HB oligomers caused the dissociation of PhaR from PhaR-DNA complexes, but native PHA granules, which were covered with PhaP or other nonspecific proteins, did not cause the dissociation. These results suggest that PhaR is able to sense both the onset of PHA synthesis and the enlargement of the granules through direct binding to PHA. However, free PhaR is probably unable to sense the mature PHA granules which are already covered sufficiently with PhaP and/or other proteins. An in vitro expression experiment revealed that phaP expression was repressed by the addition of PhaR and was derepressed by the addition of P(3HB). Based on these findings, we present here a possible model accounting for the PhaR-mediated mechanism of PHA synthesis. Widespread distribution of PhaR homologs in short-chain-length PHA producing bacteria suggests a common and important role of PhaR-mediated regulation of PHA synthesis. PMID- 12081973 TI - Burkholderia thailandensis E125 harbors a temperate bacteriophage specific for Burkholderia mallei. AB - Burkholderia thailandensis is a nonpathogenic gram-negative bacillus that is closely related to Burkholderia mallei and Burkholderia pseudomallei. We found that B. thailandensis E125 spontaneously produced a bacteriophage, termed phiE125, which formed turbid plaques in top agar containing B. mallei ATCC 23344. We examined the host range of phiE125 and found that it formed plaques on B. mallei but not on any other bacterial species tested, including B. thailandensis and B. pseudomallei. Examination of the bacteriophage by transmission electron microscopy revealed an isometric head and a long noncontractile tail. B. mallei NCTC 120 and B. mallei DB110795 were resistant to infection with phiE125 and did not produce lipopolysaccharide (LPS) O antigen due to IS407A insertions in wbiE and wbiG, respectively. wbiE was provided in trans on a broad-host-range plasmid to B. mallei NCTC 120, and it restored LPS O-antigen production and susceptibility to phiE125. The 53,373-bp phiE125 genome contained 70 genes, an IS3 family insertion sequence (ISBt3), and an attachment site (attP) encompassing the 3' end of a proline tRNA (UGG) gene. While the overall genetic organization of the phiE125 genome was similar to lambda-like bacteriophages and prophages, it also possessed a novel cluster of putative replication and lysogeny genes. The phiE125 genome encoded an adenine and a cytosine methyltransferase, and purified bacteriophage DNA contained both N6-methyladenine and N4-methylcytosine. The results presented here demonstrate that phiE125 is a new member of the lambda supergroup of Siphoviridae that may be useful as a diagnostic tool for B. mallei. PMID- 12081974 TI - In vivo and in vitro studies of Bacillus subtilis ferrochelatase mutants suggest substrate channeling in the heme biosynthesis pathway. AB - Ferrochelatase (EC 4.99.1.1) catalyzes the last reaction in the heme biosynthetic pathway. The enzyme was studied in the bacterium Bacillus subtilis, for which the ferrochelatase three-dimensional structure is known. Two conserved amino acid residues, S54 and Q63, were changed to alanine by site-directed mutagenesis in order to detect any function they might have. The effects of these changes were studied in vivo and in vitro. S54 and Q63 are both located at helix alpha3. The functional group of S54 points out from the enzyme, while Q63 is located in the interior of the structure. None of these residues interact with any other amino acid residues in the ferrochelatase and their function is not understood from the three-dimensional structure. The exchange S54A, but not Q63A, reduced the growth rate of B. subtilis and resulted in the accumulation of coproporphyrin III in the growth medium. This was in contrast to the in vitro activity measurements with the purified enzymes. The ferrochelatase with the exchange S54A was as active as wild-type ferrochelatase, whereas the exchange Q63A caused a 16-fold reduction in V(max). The function of Q63 remains unclear, but it is suggested that S54 is involved in substrate reception or delivery of the enzymatic product. PMID- 12081975 TI - Microarray analysis of the Mycobacterium tuberculosis transcriptional response to the acidic conditions found in phagosomes. AB - We used microarrays and real-time reverse transcription-PCR to analyze the global transcriptional response of Mycobacterium tuberculosis to low pH in vitro, which may mimic an environmental signal encountered by phagocytosed mycobacteria. Eighty-one genes were differentially expressed >1.5-fold, including many involved in fatty acid metabolism. The most highly induced genes showed homology with nonribosomal peptide synthetases/polyketide synthases. PMID- 12081976 TI - Differential spectrum of mutations that activate the Escherichia coli bgl operon in an rpoS genetic background. AB - The bgl promoter is silent in wild-type Escherichia coli under standard laboratory conditions, and as a result, cells exhibit a beta-glucoside-negative (Bgl-) phenotype. Silencing is brought about by negative elements that flank the promoter and include DNA structural elements and sequences that interact with the nucleoid-associated protein H-NS. Mutations that confer a Bgl+ phenotype arise spontaneously at a detectable frequency. Transposition of DNA insertion elements within the regulatory locus, bglR, constitutes the major class of activating mutations identified in laboratory cultures. The rpoS-encoded sigmaS, the stationary-phase sigma factor, is involved in both physiological as well as genetic changes that occur in the cell under stationary-state conditions. In an attempt to see if the rpoS status of the cell influences the nature of the mutations that activate the bgl promoter, we analyzed spontaneously arising Bgl+ mutants in rpoS+ and rpoS genetic backgrounds. We show that the spectrum of activating mutations in rpoS cells is different from that in rpoS+ cells. Unlike rpoS+ cells, where insertions in bglR are the predominant activating mutations, mutations in hns make up the majority in rpoS cells. The physiological significance of these differences is discussed in the context of survival of natural populations of E. coli. PMID- 12081977 TI - Activity of Sinorhizobium meliloti NodAB and NodH enzymes on thiochitooligosaccharides. AB - Rhizobium bacteria synthesize signal molecules called Nod factors that elicit responses in the legume root during nodulation. Nod factors, modified N-acylated beta-(1,4)-N-acetylglucosamine, are synthesized by the nodulation (nod) gene products. We tested the ability of three Sinorhizobium meliloti nod gene products to modify Nod factor analogs with thio linkages instead of O-glycosidic bonds in the oligosaccharide backbone. PMID- 12081978 TI - The fixA and fixB genes are necessary for anaerobic carnitine reduction in Escherichia coli. AB - In Escherichia coli, the use of carnitine as a terminal electron acceptor depends on a functional caiTABCDE operon. It had been suggested that the adjacent but divergent fixABCX operon is also required for carnitine metabolism, perhaps to provide electrons for carnitine reduction. We have constructed E. coli fixA and fixB mutants and find that they are unable to reduce carnitine to gamma butyrobetaine under anaerobic conditions. PMID- 12081980 TI - What is "the matter" with restenosis in 2002? PMID- 12081981 TI - Disrobing the emperor (heart) without destroying the dignity of super-normality. PMID- 12081982 TI - Statin therapy: beyond cholesterol lowering and antiinflammatory effects. PMID- 12081983 TI - Management of effusive and constrictive pericardial heart disease. PMID- 12081984 TI - Serotonin transporter gene polymorphism and myocardial infarction: Etude Cas Temoins de l'Infarctus du Myocarde (ECTIM). AB - BACKGROUND: Depression is a risk factor for myocardial infarction (MI). Selective serotonin reuptake inhibitors reduce this risk. The site of action is the serotonin transporter (SLC6A4), which is expressed in brain and blood cells. A functional polymorphism in the promoter region of the SLC6A4 gene has been described. This polymorphism may be associated with the risk of MI. METHODS AND RESULTS: The SLC6A4 polymorphism has been investigated by polymerase chain reaction in 671 male patients with MI and in 688 controls from the Etude Cas Temoins de l'Infarctus du Myocarde (ECTIM) multicentric study. Percentages for LL, LS, and SS genotypes were 35.5%, 45.4%, and 19.1%, respectively, for cases versus 28.1%, 49.1%, and 22.8%, respectively, for controls. S allele frequency was 41.8% and 47.4% for cases and controls, respectively. After adjustment for age and center by using multivariable logistic regression, the odds ratio for MI associated with the LL genotype was 1.40 (95% CI 1.11 to 1.76, P=0.0047). CONCLUSIONS: The LL genotype of the SLC6A4 polymorphism is associated with a higher risk of MI. This could be attributable to the effect of the polymorphism on serotonin-mediated platelet activation or smooth muscle cell proliferation or on other risk factors, such as depression or response to stress. PMID- 12081985 TI - Early resolution of ST-segment elevation correlates with myocardial salvage assessed by Tc-99m sestamibi scintigraphy in patients with acute myocardial infarction after mechanical or thrombolytic reperfusion therapy. AB - BACKGROUND: Early resolution of ST-segment elevation is an indicator of final infarct size and clinical outcomes. Whether this correlation is an expression of initial infarct characteristics or degree of myocardial salvage achieved with reperfusion therapy is unclear. METHODS AND RESULTS: We prospectively included 243 patients with acute myocardial infarction (AMI) treated with either coronary stenting (122 patients) or thrombolysis (121 patients). Serial 12-lead electrocardiograms (ECG) were performed at baseline and 90 minutes after initiation of therapy. ST-segment resolution was defined as complete (> or =70%), partial (<70% to 30%), or no resolution (<30%). Paired technetium-99m sestamibi scintigraphic studies were performed to calculate the initial perfusion defect, final infarct size, and the proportion of initial defect salvaged with reperfusion (salvage index). ST-segment resolution correlated significantly with salvage index (P=0.008), final infarct size (P<0.001), and 6-month mortality (P=0.03). In the groups with complete, partial, and no resolution, salvage index was 0.54+/-0.32, 0.39+/-0.36, and 0.33+/-0.60; final infarct size was 12.5+/ 12.0%, 20.0+/-13.9%, and 22.7+/-19.4% of the left ventricle; 6-month mortality was 2.4%, 6.2%, and 12.8%, respectively. After adjustment for baseline characteristics, ST-segment resolution was the second strongest predictor of salvage index (P=0.007) after the type of reperfusion, stenting, or thrombolysis (P=0.001); it was greater after stenting than after thrombolysis (P<0.001). CONCLUSION: Early resolution of ST-segment elevation in surface ECG correlates with myocardial salvage as assessed by scintigraphy in patients with AMI after reperfusion therapy. These data provide an explanation for the favorable prognostic value of the ST-segment resolution and support the use of this parameter to compare the efficacy of different reperfusion strategies. PMID- 12081986 TI - Coronary pressure measurement after stenting predicts adverse events at follow up: a multicenter registry. AB - BACKGROUND: Coronary stenting is associated with a restenosis rate of 15% to 20% at 6-month follow-up, despite optimum angiographic stent implantation. In this multicenter registry, we investigated the relation between optimum physiological stent implantation as assessed by poststent fractional flow reserve (FFR) and outcome at 6 months. METHODS AND RESULTS: In 750 patients, coronary pressure measurement at maximum hyperemia was performed after angiographically apparently satisfactory stent implantation. Poststenting FFR was calculated and related to major adverse events (including need for repeat target vessel revascularization) at 6 months. In 76 patients (10.2%), at least 1 adverse event occurred. Five patients died, 19 experienced myocardial infarction, and 52 underwent at least 1 repeat target vessel revascularization. By multivariate analysis, FFR immediately after stenting was the most significant independent variable related to all types of events. In 36% of the patients, FFR normalized (>0.95), and event rate was 4.9% in that group. In 32% of the patients, poststent FFR was between 0.90 and 0.95, and event rate was 6.2%. In 32% of patients, poststent FFR was <0.90, and event rate was 20.3%. In 6% of the patients, FFR was <0.80, and event rate was 29.5% (P<0.001). CONCLUSIONS: FFR after stenting is a strong independent predictor of outcome at 6 months. PMID- 12081987 TI - Omapatrilat reduces pulse pressure and proximal aortic stiffness in patients with systolic hypertension: results of the conduit hemodynamics of omapatrilat international research study. AB - BACKGROUND: Increased pulse pressure, an indicator of conduit vessel stiffness, is a strong independent predictor of cardiovascular events in hypertensive cohorts, which suggests that reduction of conduit vessel stiffness may be desirable in hypertension. METHODS AND RESULTS: We assessed changes in pulse pressure and conduit vessel stiffness in a 12-week double-blind, randomized clinical trial that compared monotherapy with the ACE inhibitor enalapril 40 mg daily (n=87) versus the vasopeptidase (dual ACE and neutral endopeptidase) inhibitor omapatrilat 80 mg daily (n=80) in patients with systolic hypertension. Patients were withdrawn from antihypertensive medications 1 to 2 weeks before enrollment, and systolic pressure was confirmed to be > or =160 mm Hg. With the use of calibrated tonometry and pulsed Doppler, pulsatile hemodynamics were assessed before randomization and at 12 weeks. Characteristic impedance (Z(c)), a direct measure of the stiffness of the central aorta, was calculated from the ratio of changes in carotid pressure and aortic flow in early systole. Omapatrilat compared with enalapril produced greater reductions in peripheral ( 8.2+/-12.2 versus -4.0+/-12.2 mm Hg, P<0.05) and central (-10.2+/-16.2 versus 3.2+/-16.9 mm Hg, P<0.01) pulse pressures and Z(c) (237+/-83 to 208+/-70 versus 225+/-87 to 231+/-94 dyne x s/cm(5), P<0.001); the latter remained significant (P<0.05) after adjusting for change in mean pressure. CONCLUSIONS: Greater reductions in pulse pressure and Z(c) in hypertensive subjects treated with omapatrilat compared with enalapril suggest that aortic stiffness is maintained by specific, partially reversible mechanisms and underscore a potential role for pharmacological modulation of natriuretic peptides in the treatment of hypertension. PMID- 12081988 TI - Statin therapy, cardiovascular events, and total mortality in the Heart and Estrogen/Progestin Replacement Study (HERS). AB - BACKGROUND: Although effects of statins on cardiovascular outcomes are well established in men, fewer data exist for women. Furthermore, the effects of statins plus hormone replacement therapy (HRT) on cardiovascular outcomes are uncertain. METHODS AND RESULTS: We examined statin use, cardiovascular events, and total mortality in the Heart and Estrogen/progestin Replacement Study (HERS), a randomized clinical trial of estrogen plus progestin versus placebo in postmenopausal women with heart disease (n=2763). A nonrandomized comparison of statin users and nonusers revealed lower rates of the primary outcome, nonfatal myocardial infarction or coronary heart disease death (relative hazard [RH]=0.79, 95% confidence intervals [CI] 0.63 to 0.99, P=0.04), and total mortality (RH=0.67, 95% CI 0.51 to 0.87, P=0.003). Rates of venous thromboembolic events were also lower among statin users (RH=0.45, 95% CI 0.23 to 0.88, P=0.02). HRT resulted in a significant increase in early risk for primary events in women who did not use statins (RH=1.75, 95% CI 1.02 to 3.03, P=0.04) but not in statin users (RH=1.34, 95% CI 0.63 to 2.86, P=0.45). Adjustment for postrandomization statin use showed no effect of HRT on risk for the primary outcome (RH=0.96, 95% CI 0.77 to 1.29; P=0.72). CONCLUSIONS: In HERS, statin use was associated with lower rates of cardiovascular events, venous thromboembolic events, and total mortality. These data provide strong support for statin use in eligible women with coronary disease. PMID- 12081989 TI - Polymorphism in the 5'-flanking region of human glutamate-cysteine ligase modifier subunit gene is associated with myocardial infarction. AB - BACKGROUND: Human glutamate-cysteine ligase (GCL) is a rate-limiting enzyme for the synthesis of glutathione that plays a crucial role in antioxidant defense mechanisms in most mammalian cells, including vascular cells. Oxidants transcriptionally upregulate GCL genes for glutathione synthesis, providing a protective mechanism against oxidative stress-induced cellular dysfunction. This study examined the hypothesis that variation in the GCL genes may be associated with coronary artery disease in which oxidative stress plays a pathogenetic role. METHODS AND RESULTS: We searched for the common variants in the 5'-flanking region of the GCL modifier subunit (GCLM) gene in patients with myocardial infarction (MI). We found a polymorphism (-588C/T) in which the T allele showed lower promoter activity (40% to 50% of C allele) in response to oxidants in the luciferase reporter gene assay. Allele frequencies were determined by polymerase chain reaction-based analysis of restriction fragment length polymorphism in 429 patients with MI and 428 control subjects (as defined by angiography) in Kumamoto Prefecture, Japan. The frequency of the T polymorphism was significantly higher in the MI group than in the control group (CT and TT genotypes: 31.5% in MI group versus 19.2% in control group; P<0.001). In multiple logistic regression analysis, the T polymorphism was a risk factor for MI independent of traditional coronary artery disease risk factors (odds ratio, 1.98; 95% confidence interval, 1.38 to 2.83; P<0.001). CONCLUSIONS: These findings suggest that the -588T polymorphism of the GCLM gene may suppress GCLM gene induction in response to oxidants and that it is a genetic risk factor for MI. PMID- 12081990 TI - Morphological predictors of restenosis after coronary stenting in humans. AB - BACKGROUND: Experimental studies suggest that arterial injury and inflammation lead to increased neointimal growth after stenting. Despite the increased use of coronary stents in humans, there are only limited pathological data on the morphological features of in-stent restenosis. METHODS AND RESULTS: Detailed histology was performed on 116 stents, implanted > or =90 days in 87 coronary arteries, from 56 patients (mean age, 59+/-13 years). The mean duration of stent implant was 10 months. In-stent restenosis was defined as a stent area stenosis of >75%. Lumen area increased as stent area increased (r2=0.27, P=0.0001), but there was a much stronger correlation between stent area and neointimal area (r2=0.70, P<0.0001). Arterial medial fracture was associated with a 29% increase (P<0.01) in neointimal thickness compared with arteries with an intact media. Neointimal thickness (P=0.0001), inflammatory cell density (P<0.0001), and neointimal vascular channel density (P<0.0001) were greater when struts were in contact with a ruptured arterial media compared with fibrous plaque or an intact fibrous cap. Stent strut penetration into a lipid core was associated with increased neointimal thickness (P=0.04) and inflammatory cell density (P=0.03). Neointimal inflammatory cell content was 2.4-fold greater in stents with restenosis versus no restenosis, and inflammation was associated with increased neoangiogenesis. CONCLUSIONS: Coronary stenting that is accompanied by medial damage or penetration of the stent into a lipid core induces increased arterial inflammation, which is associated with increased neointimal growth. These data suggest the use of stenting strategies that reduce inflammation and neoangiogenesis to reduce the incidence of restenosis. PMID- 12081991 TI - Abciximab attenuates coronary microvascular endothelial dysfunction after coronary stenting. AB - BACKGROUND: Platelet glycoprotein IIb/IIIa receptor blockade with abciximab decreases ischemic events after percutaneous coronary intervention (PCI); however, the mechanism of this benefit has not been fully elucidated. The present study was designed to assess endothelium-dependent vasomotion after coronary stenting and to determine if abciximab alters this response. METHODS AND RESULTS: The study group consisted of 48 patients (59+/-10 years of age) with discrete coronary stenoses who underwent stenting alone (n=28) or stenting plus abciximab (n=20). A control group consisted of 31 additional patients who had vasomotor testing on a non-PCI vessel. Coronary blood flow (CBF) was measured (0.014-inch Doppler wire) 30 minutes after uncomplicated PCI and in response to the intracoronary infusion of acetylcholine (Ach) (10(-7), 10(-6) mol/L Ach) and adenosine (24 microg). Ach-mediated increase in CBF was impaired after stent insertion when compared with the control group (41+/-52% versus 70+/-48%; P<0.05). The stenting plus abciximab group demonstrated a superior CBF response to Ach compared with the stenting alone group (83+/-93% versus 41+/-52%; P<0.05), with no difference between groups in the peak flow or percent change in flow to adenosine. By multivariate analysis, concomitant administration of abciximab was strongly predictive of the change in CBF to Ach (P<0.005). CONCLUSIONS: Abciximab preserves the CBF response to Ach after coronary stenting. The preservation of microvascular endothelial function may help explain the beneficial clinical effect of this agent in patients undergoing PCI. PMID- 12081992 TI - Long-term clinical and angiographic follow-up after coronary stent placement in native coronary arteries. AB - BACKGROUND: Although coronary stents have been proved effective in reducing clinical cardiac events for up to 3 to 5 years, longer term clinical and angiographic outcomes have not yet been fully clarified. METHODS AND RESULTS: To evaluate longer term (7 to 11 years) outcome, clinical and angiographic follow-up information was analyzed in 405 patients with successful stenting in native coronary arteries. Primary or secondary stabilization, which was defined as freedom from death, coronary artery bypass grafting, and target lesion percutaneous coronary intervention (TL-PCI) during the 14 months after the initial procedure or after the last TL-PCI, was achieved in 373 patients (92%) overall. Only 7 patients (1.7%) underwent TL-PCI more than twice. After the initial 14-month period, freedom from TL-PCI reached a plateau at 84.9% to 80.7% over 1 to 8 years. However, quantitative angiographic analysis in 179 lesions revealed a triphasic luminal response characterized by an early restenosis phase until 6 months, an intermediate-term regression phase from 6 months to 3 years, and a late renarrowing phase beyond 4 years. Minimal luminal diameter in 131 patients with complete serial data were 2.62+/-0.4 mm immediately after stenting, 2.0+/-0.49 mm at 6 months, 2.19+/-0.49 mm at 3 years, and 1.85+/-0.56 mm beyond 4 years (P<0.0001). CONCLUSIONS: The efficacy and safety of coronary stenting seemed to be clinically sustained at 7 to 11 years of follow-up. However, late luminal renarrowing beyond 4 years was common, which demonstrates the need for further follow-up. PMID- 12081993 TI - Assessment of diastolic function with Doppler tissue imaging to predict genotype in preclinical hypertrophic cardiomyopathy. AB - BACKGROUND: Unexplained left ventricular hypertrophy (LVH) is considered diagnostic of hypertrophic cardiomyopathy (HCM) but fails to identify all genetically affected individuals. Altered diastolic function has been hypothesized to represent an earlier manifestation of HCM before the development of LVH; however, data regarding the clinical utility of imaging techniques that assess this parameter are limited. METHODS AND RESULTS: Echocardiographic studies including Doppler tissue imaging (DTI) were performed in a genotyped HCM population with beta-myosin heavy chain (beta -MHC) mutations. Genotype (+) individuals with LVH (G+/LVH+; n=18) and genotype (+) individuals without LVH (G+/LVH-; n=18) were compared with normal control subjects (n=36). Left ventricular ejection fraction (EF) was significantly higher in both genotype (+) groups (75+/-5% and 71+/-6%, respectively, versus 64+/- 5% in control subjects; P<0.0001). Mean early diastolic myocardial velocities (Ea) were significantly lower in both genotype (+) subgroups, irrespective of LVH (P<0.02). However, there was substantial overlap in Ea velocities between the G+/LVH- and control groups. An Ea velocity of < or =13.5 cm/s had 86% specificity and 75% sensitivity for identifying genotype-positive subjects. The combination of EF > or =68% and Ea velocity <15 cm/s was 100% specific and 44% sensitive in predicting affected genotype. CONCLUSIONS: Abnormalities of diastolic function assessed by Doppler tissue imaging precede the development of LVH in individuals with HCM caused by beta -MHC mutations. Although Ea velocity alone was not sufficiently sensitive as a sole diagnostic criterion, the combination of Ea velocity and EF was highly predictive of affected genotype in individuals without overt manifestations of HCM. PMID- 12081994 TI - Electrical connections between pulmonary veins: implication for ostial ablation of pulmonary veins in patients with paroxysmal atrial fibrillation. AB - BACKGROUND: Electrical disconnection of the myocardial extensions into arrhythmogenic pulmonary veins (PVs) is recognized as a curative technique for paroxysmal atrial fibrillation (AF). However, the presence of electrical connections between the PVs, which may make achievement of PV disconnection difficult, has not been systematically evaluated. METHODS AND RESULTS: Forty-nine consecutive patients with drug-resistant AF underwent ostial radiofrequency (RF) catheter ablation of arrhythmogenic PVs with foci triggering AF. Pacing from inside the targeted PV was performed after each RF delivery to identify the left atrial exit site of the residual venoatrial conduction. Successful PV disconnection was defined as achieving elimination of the PV potentials during sinus rhythm or left atrial pacing, and the loss of left atrial conduction during intra-PV pacing. A total of 112 arrhythmogenic PVs were identified. PV disconnection was achieved with 10+/-6.1 minutes of RF delivery to the ostia of 101 targeted PVs. In 7 left superior (LS) PVs from 7 patients (14%), the earliest atrial activity was recorded from the left inferior (LI) PV ostium during intra LSPV pacing after 11+/-4.7 minutes of RF delivery to the LSPV ostium. Disconnection of these LSPVs was achieved by LIPV disconnection. In the remaining 4 PVs from 4 patients, PV disconnection could not be achieved. CONCLUSIONS: Fourteen percent of the patients had electrical connections between contiguous PVs. In these patients, ostial ablation of an untargeted PV was required for successful targeted PV disconnection. PMID- 12081995 TI - Sequential follow-up results of catheter intervention for coronary artery lesions after Kawasaki disease: quantitative coronary artery angiography and intravascular ultrasound imaging study. AB - BACKGROUND: The purpose of this study was to assess the sequential follow-up results of catheter intervention in Kawasaki disease by use of quantitative coronary angiography (QCA) and intravascular ultrasound imaging. METHODS AND RESULTS: Catheter intervention was performed on 23 stenotic lesions in 22 patients (aged 2 to 24 years). Percutaneous balloon angioplasty (PBA) was performed in 4 patients, stent implantation in 7, percutaneous transluminal coronary rotational ablation (PTCRA) in 10, and a combination of PTCRA with stent implantation in 2. A total of 21 lesions (91%) were successfully dilated by catheter intervention without major or minor complications. One patient immediately underwent coronary artery bypass grafting (CABG) surgery because stent implantation failed to resolve his lesion. At 4 to 6 months after catheter intervention, 2 restenotic lesions (9%) were detected by QCA in 2 patients who had undergone PBA, and these patients subsequently underwent CABG surgery. In 6 months to 3 years after catheter intervention, no patients showed evidence of ischemic findings. At 3 to 4 years after catheter intervention, QCA and intravascular ultrasound studies were performed on 15 lesions in 14 patients. Two restenotic lesions (13%) were detected by QCA in 2 patients. One of the 2 had stent implantation and underwent CABG surgery, and the other had undergone PTCRA and underwent re-PTCRA. Thirteen patients demonstrated no ischemic findings at 3 to 8 years after catheter intervention. CONCLUSION: Catheter intervention for Kawasaki disease can be accomplished and can be effective in the short term, but the long-term efficacy should be verified by further study. PMID- 12081996 TI - Cardiac nitric oxide synthase 1 regulates basal and beta-adrenergic contractility in murine ventricular myocytes. AB - BACKGROUND: Evidence indicates that myocardial NO production can modulate contractility, but the source of NO remains uncertain. Here, we investigated the role of a type 1 NO synthase isoform (NOS1), which has been recently localized to the cardiac sarcoplasmic reticulum, in the regulation of basal and beta adrenergic myocardial contraction. METHODS AND RESULTS: Contraction was assessed in left ventricular myocytes isolated from mice with NOS1 gene disruption (NOS1( /-) mice) and their littermate controls (NOS1(+/+) mice) at 3 stimulation frequencies (1, 3, and 6 Hz) in basal conditions and during beta-adrenergic stimulation with isoproterenol (2 nmol/L). In addition, we examined the effects of acute specific inhibition of NOS1 with vinyl-L-N-5-(1-imino-3-butenyl)-L ornithine (L-VNIO, 500 micromol/L). NOS1((-/-)) myocytes exhibited greater contraction at all frequencies (percent cell shortening at 6 Hz, 10.7+/-0.92% in NOS1(-/-) myocytes versus 7.21+/-0.8% in NOS1(+/+) myocytes; P<0.05) with a flat frequency-contraction relationship. Time to 50% relaxation was increased in NOS1( /-) myocytes at all frequencies (at 6 Hz, 26.53+/-1.4 ms in NOS1(-/-) myocytes versus 21.27+/-1.3 ms in NOS1(+/+) myocytes; P<0.05). L-VNIO prolonged time to 50% relaxation at all frequencies (at 6 Hz, 21.28+/-1.7 ms in NOS1(+/+) myocytes versus 26.45+/-1.4 ms in NOS1(+/+)+L-VNIO myocytes; P<0.05) but did not significantly increase basal contraction. However, both NOS1(-/-) myocytes and NOS1(+/+) myocytes treated with L-VNIO showed a greatly enhanced contraction in response to beta-adrenergic stimulation (percent increase in contraction at 6 Hz, 25.2+/-10.8 in NOS1(+/+) myocytes, 68.2+/-11.2 in NOS1(-/-) myocytes, and 65.1+/ 13.2 in NOS1(+/+)+L-VNIO myocytes; P<0.05). CONCLUSIONS: NOS1 disruption enhances basal contraction and the inotropic response to beta-adrenergic stimulation in murine ventricular myocytes. These findings indicate that cardiac NOS1-derived NO plays a significant role in the autocrine regulation of myocardial contractility. PMID- 12081998 TI - Influence of C3 deficiency on atherosclerosis. AB - BACKGROUND: The influence of complement activation on atherosclerosis is not well understood. The purpose of this study was to examine the effects of C3 deficiency on the extent and phenotype of atherosclerosis. METHODS AND RESULTS: Aortic atherosclerosis was analyzed in low-density lipoprotein receptor (ldlr)/C3 deficient mice (ldlr(-/-)C3(-/-)) and ldlr(-/-)C3(+/-) littermate control mice after 15 weeks on a 1.25% (wt/wt) cholesterol diet. Serum lipoprotein profiles and immunoglobulin levels were not significantly different between the 2 experimental groups. The lipid-positive en face lesional area in thoracic and abdominal aorta was greater in C3-deficient mice than in control mice (3.9% versus 2.1%, median, P=0.0076). Similarly, the lipid-positive area in aortic arch sections was greater in C3-deficient mice than in controls (0.04 mm2 versus 0.02 mm2, median, P=0.0089). Analysis of aortic arch sections showed greater lesional macrophage content in C3-deficient versus control mice (8.24+/-1.36% versus 5.9+/ 1.63% intimal area, mean+/-SEM, P=0.003), less smooth muscle cell content in C3 deficient versus control mice (0.06+/-0.05% versus 0.92+/-0.32% intimal area, mean+/-SEM, P<0.0001), and less collagen content in C3-deficient versus control mice (0.52+/-1.26% versus 11+/-10.43% intimal area, mean+/-SEM, P=0.008). CONCLUSIONS: The maturation of atherosclerotic lesions beyond the foam cell stage is strongly dependent on an intact complement system. PMID- 12081997 TI - Statin therapy accelerates reendothelialization: a novel effect involving mobilization and incorporation of bone marrow-derived endothelial progenitor cells. AB - BACKGROUND: Primary and secondary prevention trials suggest that statins possess favorable effects independent of cholesterol reduction. We investigated whether statin therapy may also accelerate reendothelialization after carotid balloon injury. METHODS AND RESULTS: Simvastatin treatment in 34 male Sprague-Dawley rats accelerated reendothelialization of the balloon-injured arterial segments (reendothelialized area at 2 weeks, 12.3+/-1.8 versus 5.4+/-1.1 mm2, P< 0.01) and resulted in a dose-dependent (0.2 or 1 mg/kg IP) significant reduction in neointimal thickening at 2, 3, and 4 weeks compared with saline-injected controls (n=18). To elucidate the mechanism, we investigated the contribution of bone marrow-derived endothelial progenitor cells (EPCs) by bone marrow transplantation from Tie2/lacZ mice to background mice or nude rats. X-gal staining of mouse carotid artery specimens revealed a 2.9-fold increase in the number of beta-gal positive cells per square millimeter appearing on the carotid artery luminal surface at 2 weeks, and double-fluorescence immunohistochemistry disclosed a significant 5-fold increase in the number of double-positive cells (beta-gal, isolectin B4) on the luminal surface in carotid arteries of statin-treated nude rats (20+/-3 versus 4+/-1 cells/mm surface length, P<0.005). Statins increased circulating rat EPCs (2.4-fold at 2 weeks and 2.5-fold at 4 weeks, P<0.001) and induced adhesiveness of cultured human EPCs by upregulation of the integrin subunits alpha5, beta1, alpha(v), and beta5 of human EPCs as shown by reverse transcription-polymerase chain reaction and fluorescence-activated cell sorting. CONCLUSIONS: These findings establish additional mechanisms by which statins may specifically preempt disordered vascular wall pathology and constitute physiological evidence that EPC mobilization represents a functionally relevant consequence of statin therapy. PMID- 12081999 TI - S-nitroso human serum albumin treatment reduces ischemia/reperfusion injury in skeletal muscle via nitric oxide release. AB - BACKGROUND: Peroxynitrite generated from nitric oxide (NO) and superoxide (O2-) contributes to ischemia/reperfusion (I/R) injury. Feedback inhibition of endothelial NO synthase by NO may inhibit O2- production generated also by endothelial NO synthase at diminished local L-arginine concentrations accompanying I/R. METHODS AND RESULTS: During hindlimb I/R (2.5 hours/2 hours), in vivo NO was monitored continuously (porphyrinic sensor), and high-energy phosphates, reduced and oxidized glutathione (chromatography), and I/R injury were measured intermittently. Rabbits receiving human serum albumin (HSA) (controls) were compared with those receiving S-nitroso human serum albumin (S-NO HSA) beginning 30 minutes before reperfusion for 1 hour or 30 minutes before ischemia for 3.5 hours (0.1 micromol x kg(-1) x h(- 1)). The onset of ischemia led to a rapid increase of NO from its basal level (50+/-12 nmol/L) to 120+/-20 and 220+/-15 nmol/L in the control and S-NO-HSA-treated groups, respectively. In control animals, NO dropped below basal levels at the end of ischemia and to undetectable levels (<1 nmol/L) during reperfusion. In S-NO-HSA-treated animals, maximal NO levels never decreased below basal concentration and on reperfusion were 100+/-15 nmol/L (S-NO-HSA preischemia group, 175+/-15 nmol/L). NO supplementation by S-NO-HSA led to partial and in the preischemia group to total preservation of high-energy phosphates and glutathione status in reperfused muscle (eg, preischemia groups: ATP, 30.23+/-5.02 micromol/g versus control, 15.75+/-4.33 micromol/g, P<0.0005; % oxidized glutathione, 4.49+/- 1.87% versus control, 22.84+/-6.39%, P<0.0001). S-NO-HSA treatment in all groups led to protection from vasoconstriction and reduced edema formation after reperfusion (eg, preischemia groups: interfiber area, 12.94+/-1.36% versus control, 27.83+/ 1.95%, P< 0.00001). CONCLUSIONS: Long-lasting release of NO by S-NO-HSA provides significant protection of skeletal muscle from I/R injury. PMID- 12082000 TI - Sensitivity to apoptosis signal, clearance rate, and ultrastructure of fas ligand induced apoptosis in in vivo adult cardiac cells. AB - BACKGROUND: Sensitivity to apoptotic signals, the clearance rate of apoptosis, and the apoptotic ultrastructure have not been studied in cells of the in vivo adult heart. METHODS AND RESULTS: To minimize the systemic influence, soluble Fas ligand was injected directly into in vivo rat hearts and livers (as the control) at concentrations of 0, 0.5, 2, and 5 microg/mL (groups C, F0.5, F2, and F5). Apoptotic cardiomyocytes and apoptotic noncardiomyocytes of the heart were identified with similar incidences only in F5. Their incidences peaked at 12 hours after injection (2.0+/-0.09% in cardiomyocytes) and diminished markedly 24 hours later. Caspase-3 was activated only in F5. Boc-Asp-fmk, a pancaspase inhibitor, inhibited apoptosis, suggesting that the apoptosis sensitivity was regulated upstream of caspase-3. Apoptotic noncardiomyocytes showed typical ultrastructure. In addition to the typical ultrastructure, such as cellular shrinkage, chromatin condensation, and apoptotic bodies, however, apoptotic cardiomyocytes showed unique features: doughnut-like, but not half-moon- or crescent-like, chromatin condensation; frequent plasma membrane rupture even during the early stage; condensed mitochondria with wrinkled cristae inside; the appearance of cytoplasmic lipid-like droplets; and myofibrillar derangement. In the livers, typical apoptosis was induced in hepatocytes and nonhepatocytes of the liver even in the F0.5 group, which were cleared 24 hours later. CONCLUSIONS: Compared with liver cells, cardiomyocytes as well as noncardiomyocytes of the heart are more resistant against the apoptotic signal, but the clearance is similarly rapid (within 24 hours). The ultrastructure of apoptotic cardiomyocytes is unique. These findings provide new insights into the dynamics of cell death in the heart. PMID- 12082001 TI - Role of cyclic guanosine monophosphate in late preconditioning in conscious rabbits. AB - BACKGROUND: Although NO has been shown to serve both as the trigger and the mediator of the late phase of ischemic preconditioning (PC), it is unknown whether NO acts via activation of soluble guanylate cyclase (sGC). The objective of this study was to investigate the role of sGC in late PC in conscious rabbits using the selective sGC inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ). METHODS AND RESULTS: A total of 172 conscious rabbits were used. When nonpreconditioned rabbits were subjected to a sequence of 4-minute coronary occlusion/4-minute reperfusion cycles, myocardial cyclic guanosine monophosphate (cGMP) levels increased significantly at the end of the third and sixth occlusions. In rabbits preconditioned 24 hours earlier (on day 1) with six occlusion/reperfusion cycles, myocardial cGMP levels on day 2 were significantly higher than in nonpreconditioned rabbits even before ischemia but did not increase further during a second sequence of 4-minute occlusion/reperfusion cycles. Administration of ODQ before the six occlusion/reperfusion cycles on day 1 did not prevent the development of late PC against either stunning or infarction on day 2. In contrast, administration of ODQ on day 2 completely ablated the late PC effect against both stunning and infarction. CONCLUSIONS: These results indicate that enhanced synthesis of cGMP by sGC is not necessary for ischemia to trigger a late PC effect but is required for the protection to become manifest 24 hours later. This implies that NO participates in late PC via two distinct mechanisms; ie, it triggers late PC on day 1 via a cGMP-independent mechanism and it mediates late PC on day 2 via a cGMP-dependent mechanism. PMID- 12082002 TI - Polymorphic reentrant ventricular tachycardia in the isolated rabbit heart studied by high-density mapping. AB - BACKGROUND: The role of dispersion of refractoriness and reentry for the genesis of polymorphic ventricular tachycardia (VT) has recently become emphasized. We investigated the mechanisms of polymorphic arrhythmias in a 2D preparation confining an area of prolonged refractoriness. METHODS AND RESULTS: In 16 Langendorff-perfused rabbit hearts, a sheet of left ventricular epicardium was obtained by a cryoprocedure. Enhanced spatial heterogeneity in a refractory period was created by cooling a central region (diameter=12 mm). This markedly prolonged the refractory period (by 36+/-14 ms) inside but only slightly prolonged it (by 5+/-11 ms) outside the cooled area (n=6). During a control procedure, programmed stimulation with up to 3 premature stimuli induced an episode of monomorphic VT in only 1 of 10 hearts. During regional cooling, episodes of polymorphic VT with a maximum duration of 35 seconds could be induced in all hearts. High-resolution mapping (229 electrodes) of epicardial activation revealed that polymorphic VT was caused by a functional reentrant circuit located partially within the region of prolonged refractoriness. The reentrant wavefront was continuously shifting along the border of the cooled region, resulting in beat-to-beat changes in the excitation pattern. Spontaneous termination of polymorphic VT occurred either by a shift of the reentrant circuit outside the cooled region or by a block in the central common pathway during figure-of-8 reentry in the region of prolonged refractoriness. CONCLUSIONS: A shifting functional reentrant circuit was the underlying mechanism of polymorphic VT in a substrate of enhanced spatial heterogeneity of refractoriness. PMID- 12082003 TI - Defending the rationale for the two-tailed test in clinical research. PMID- 12082004 TI - Images in cardiovascular medicine. Eosinophilic pericarditis and myocarditis. PMID- 12082005 TI - Images in cardiovascular medicine. Tear, tumor, or thrombus. PMID- 12082006 TI - Ramipril and cardiovascular risk reduction. PMID- 12082007 TI - Heme oxygenase-1-mediated protection: potential role of nonheme iron-nitric oxide complexes. PMID- 12082008 TI - Plant foods have a complete amino acid composition. PMID- 12082009 TI - Off-pump versus on-pump coronary bypass surgery. PMID- 12082011 TI - Toxicogenomics: the new frontier in risk analysis. AB - Risk assessment involves the identification of potential human health hazards, the assessment of the level of exposure by humans to these hazards and the evaluation of the relationship between exposure and response in humans. Research is currently underway to improve the scientific basis of risk assessment through the incorporation of new technologies such as transgenic animals, molecular epidemiology, toxicogenomics, alternative models to animals and mechanism-based mathematical modeling into the estimation of risk in a quantitative manner. This paper briefly discusses these technologies and how each is being employed for more scientifically sound risk assessments. PMID- 12082012 TI - Human urinary carcinogen metabolites: biomarkers for investigating tobacco and cancer. AB - Measurement of human urinary carcinogen metabolites is a practical approach for obtaining important information about tobacco and cancer. This review presents currently available methods and evaluates their utility. Carcinogens and their metabolites and related compounds that have been quantified in the urine of smokers or non-smokers exposed to environmental tobacco smoke (ETS) include trans,trans-muconic acid (tt-MA) and S-phenylmercapturic acid (metabolites of benzene), 1- and 2-naphthol, hydroxyphenanthrenes and phenanthrene dihydrodiols, 1-hydroxypyrene (1-HOP), metabolites of benzo[a]pyrene, aromatic amines and heterocyclic aromatic amines, N-nitrosoproline, 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanol and its glucuronides (NNAL and NNAL-Gluc), 8 oxodeoxyguanosine, thioethers, mercapturic acids, and alkyladenines. Nitrosamines and their metabolites have also been quantified in the urine of smokeless tobacco users. The utility of these assays to provide information about carcinogen dose, delineation of exposed vs. non-exposed individuals, and carcinogen metabolism in humans is discussed. NNAL and NNAL-Gluc are exceptionally useful biomarkers because they are derived from a carcinogen- 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanone (NNK)- that is specific to tobacco products. The NNAL assay has high sensitivity and specificity, which are particularly important for studies on ETS exposure. Other useful assays that have been widely applied involve quantitation of 1-HOP and tt-MA. Urinary carcinogen metabolite biomarkers will be critical components of future studies on tobacco and human cancer, particularly with respect to new tobacco products and strategies for harm reduction, the role of metabolic polymorphisms in cancer, and further evaluation of human carcinogen exposure from ETS. PMID- 12082013 TI - Profiling and selection of genes differentially expressed in the pylorus of rat strains with different proliferative responses and stomach cancer susceptibility. AB - Rat stomach cancers induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) are widely used as a model of differentiated-type human stomach cancers. ACI/NJcl (ACI) rats show persistent and strong cell proliferation in response to gastric mucosal damage by MNNG while BUF/NacJcl (BUF) rats show transient and limited cell proliferation. This difference is considered as one of the mechanisms for the high susceptibility of ACI rats to MNNG-induced stomach carcinogenesis. To identify genes involved in the differential induction of cell proliferation, cDNA subtraction was performed using RNA isolated from the pylorus of ACI and BUF rats treated with MNNG. By the temporal patterns of their expressions, the isolated 16 genes were overviewed and clustered into groups. Expression of the genes in group 1 (such as MHC class I and class II genes and interferon-inducible genes Iigp, Mx2 and Ubd) was induced by MNNG treatment, and the genes in group 2 (such as cellular retinoic acid-binding protein II (CrabpII)) were constantly expressed regardless of MNNG treatment. Then, expression profiles among multiple rat strains were compared with the extents of induction of cell proliferation. Iigp, CrabpII and EST222005 were found to show relatively good accordance, and these three genes were considered as candidates for genes that control differential induction of cell proliferation. Presence of polymorphisms at the genomic DNA level was indicated for CrabpII and EST222005, and these two genes were considered to be better candidates than IIGP: It was shown that the temporal profiles and profiles among strains, taking advantage of animal models, are useful to select candidate genes from a collection of genes isolated by various genome-wide scanning methods. PMID- 12082014 TI - Nitric oxide mediates apoptosis induction selectively in transformed fibroblasts compared to nontransformed fibroblasts. AB - Nitric oxide (NO) mediates apoptosis induction in fibroblasts with constitutive src or induced ras oncogene expression, whereas nontransformed parental cells and revertants are not affected. This direct link between the transformed phenotype and sensitivity to NO-mediated apoptosis induction seems to be based on the recently described extracellular superoxide anion generation by transformed cells, as NO-mediated apoptosis induction in transformed cells is inhibited by extracellular superoxide dismutase (SOD), by SOD mimetics and by apocynin, an inhibitor of NADPH oxidase. Furthermore, nonresponsive nontransformed cells can be rendered sensitive for NO-mediated apoptosis induction when they are supplemented with xanthine oxidase/xanthine as an extracellular source for superoxide anions. As superoxide anions and NO readily interact in a diffusion controlled reaction to generate peroxynitrite, peroxynitrite seems to be the responsible apoptosis inducer in NO-mediated apoptosis induction. In line with this conclusion, NO-mediated apoptosis induction in superoxide anion-generating transformed cells is inhibited by the peroxynitrite scavengers ebselen and FeTPPS. Moreover, direct application of peroxynitrite induces apoptosis both in transformed and nontransformed cells, indicating that peroxynitrite is no selective apoptosis inducer per se, but that selective apoptosis induction in transformed cells by NO is achieved through selective peroxynitrite generation. The interaction of NO with target cell derived superoxide anions represents a novel concept for selective apoptosis induction in transformed cells. This mechanism may be the basis for selective apoptosis induction by natural antitumor systems (like macrophages, natural killer cells, granulocytes) that utilize NO for antitumor action. Apoptosis induction mediated by NO involves mitochondrial depolarization and is blocked by Bcl-2 overexpression. PMID- 12082015 TI - Occurrence of H-ras codon 61 CAA to AAA mutation during mouse liver tumor progression. AB - The initiating mutations of a tumor are present in each of the cancerous cells comprising the tumor. Identification and measurement of the subsequent mutations that occur during tumor progression, however, requires mutation detection in a smaller subset of the tumor cells. In this study, allele-specific competitive blocker PCR (ACB-PCR), a genotypic selection method with the sensitivity to detect a specific point mutation in the presence of a 10(5)-fold excess of wild type DNA sequence, was used to measure H-ras codon 61 CAA to AAA mutation in mouse liver tumors that did not have this mutation as an initiating event. Twenty one spontaneous or chemically induced mouse liver tumors, negative for the H-ras codon 61 CAA to AAA mutation by DNA sequencing or denaturing gradient gel electrophoresis, were analyzed for this mutation by ACB-PCR. The mutation was detected at some level in 71% of these tumors. The mutation was detected in adenomas and carcinomas more frequently (13 of 14 tumors) and at significantly higher mutant fractions than it was detected in histiocytic sarcomas (1 of 5 tumors). These data indicate that the same oncogenic point mutation that can be identified as a tumor-initiating event based on its clonal amplification in a tumor can also be present in only a small sub-population of tumor cells where the mutation must have been fixed at a later stage in tumor development. The occurrence of a mutation as a primary or secondary event probably reflects the stochastic nature of mutation and is likely to be affected by the mutation rate for each target site. PMID- 12082016 TI - High-throughput measurement of the Tp53 response to anticancer drugs and random compounds using a stably integrated Tp53-responsive luciferase reporter. AB - Human Tp53 is normally a short-lived protein. Tp53 protein is stabilized and levels are increased in response to a variety of cellular stresses, including those induced by genotoxic anticancer drugs and environmental exposures. To engineer an efficient assay based on this property, we constructed and integrated a Tp53-specific reporter system into human cancer cells, termed p53R cells. We tested a range of conventional chemotherapeutic agents as well as over 16 000 diverse small compounds. Ionizing radiation and two-thirds of conventional chemotherapeutic agents, but only 0.2% of diverse compounds activated Tp53 activity by two-fold or greater, consistent with the presumptive genotoxic activation of Tp53 function. Cytotoxicity was independent of TP53 genetic status when paired, syngeneic wild-type TP53 and TP53-null cells in culture were treated with compounds that activated Tp53. From the unbiased survey of random compounds, Tp53 activation was strongly induced by an analog of AMSA, an investigational anti-cancer agent. Tp53 was also strongly induced by an N-oxide of quinoline and by dabequine, an experimental antimalarial evaluated in humans; dabequine was reported to be negative in other screens of mutagenicity and clastogenicity but carcinogenic in animal studies. Further exploration of antimalarial compounds identified the common medicinals chloroquine, quinacrine, and amodiaquine as Tp53 inducers. Flavonoids are known to have DNA topoisomerase activity, a Tp53 inducing activity that is confirmed in the assay. A reported clinical association of Tp53 immunopositive colorectal cancers with use of the antihypertensive agents was extended by the demonstration of hydralazine and nifedipine as Tp53-inducers. p53R cells represent an efficient Tp53 functional assay to identify chemicals and other agents with interesting biologic properties, including genotoxicity. This assay may have utility in the identification of novel chemotherapeutic agents, as an adjunct in the pharmaceutical optimization of lead compounds, in the exploration of environmental exposures, and in chemical probing of the Tp53 pathway. PMID- 12082017 TI - DNA damage responses protect xeroderma pigmentosum variant from UVC-induced clastogenesis. AB - Lack of DNA polymerase eta and the attendant defect in bypass replication of pyrimidine dimers induced in DNA by ultraviolet light (UV) underlie the enhanced mutagenesis and carcinogenesis observed in xeroderma pigmentosum variant (XP-V). We investigated whether diploid XP-V fibroblasts growing in culture are also more susceptible to UV-induced clastogenesis than normal human fibroblasts (NHF). This study utilized diploid fibroblasts immortalized by the ectopic expression of human telomerase. The cell lines displayed checkpoint responses to DNA damage comparable with those measured in the parental strains. Shortly after exposure to low doses of UVC (< or =4 J/m2), XP-V cells accumulated daughter strand gaps in excess of normal controls (>25-fold). Daughter strand gaps generated in UV irradiated S phase cells are potential precursors of chromatid-type chromosomal aberrations. Nonetheless, chromatid-type chromosomal aberrations were only 1.5 to 2 times more abundant in XP-V than in NHF exposed to the same UVC dose. XP-V cells, however, displayed S phase delays at lower doses of UVC and for longer periods of time than NHF. These results support the hypothesis that aberrant DNA structures activate S phase checkpoint responses that increase the time available for postreplication repair. Alternatively, cells that cannot be properly repaired remain permanently arrested and never reach mitosis. These responses protect human cells from chromosomal aberrations, especially when other pathways, such as accurate lesion bypass, are lost. PMID- 12082018 TI - Expression profile of differentially-regulated genes during progression of androgen-independent growth in human prostate cancer cells. AB - Because of the heterogeneous nature of prostate cancer, identifying the molecular mechanisms involved during the transition from an androgen-sensitive to an androgen-independent phenotype is very complex. An LNCaP cell model that recapitulates prostate cancer progression, comprising early passage androgen sensitive (LNCaP-C33) and late passage androgen-independent (LNCaP-C81) phenotypes, would help to provide a better understanding of such molecular events. In this study, we examined the genes expressed by LNCaP-C33 and LNCaP-C81 cells using cDNA microarrays containing 1176 known genes. This analysis demonstrated that 34 genes are up-regulated and eight genes are down-regulated in androgen-independent cells. Northern blot analysis confirmed the differences identified by microarrays on several candidate genes, including c-MYC, c-MYC purine-binding transcription factor (PuF), macrophage migration inhibitory factor (MIF), macrophage inhibitory cytokine-1 (MIC-1), lactate dehydrogenase-A (LDH-A), guanine nucleotide-binding protein Gi, alpha-1 subunit (NBP), cyclin dependent kinase-2 (CDK-2), prostate-specific membrane antigen (PSM), cyclin H (CCNH), 60S ribosomal protein L10 (RPL10), 60S ribosomal protein L32 (RPL32), and 40S ribosomal protein S16 (RPS16). These differentially-regulated genes are correlated with progression of human prostate cancer and may be of therapeutic relevance as well as an aid in understanding the molecular genetic events involved in the development of this disease's hormone-refractory behavior. PMID- 12082019 TI - The growth hormone-deficient Spontaneous Dwarf rat is resistant to chemically induced mammary carcinogenesis. AB - Recent epidemiologic studies have suggested that the growth hormone (GH)/insulin like growth factor I axis plays an important role in human breast cancer. The purpose of the present study was to evaluate the function of GH in rat mammary carcinogenesis, a model that closely recapitulates human breast cancer biology. The Spontaneous Dwarf rat (SDR) arose from the Sprague-Dawley rat and harbors a mutation in its GH gene yielding undetectable levels of a severely truncated protein not capable of binding to the GH receptor. When female rats of either strain were exposed to the direct-acting carcinogen N-methyl-N-nitrosourea, all wild-type rats (n = 10) developed multiple mammary cancers (5.3/rat). In contrast, SDR rats (n = 15) developed only three cancers (0.2/rat) and these were very small (<6 mm3). In another experiment, SDRs were backcrossed with wild-type Sprague-Dawley rats and the progeny were exposed to the indirect-acting carcinogen 7,12-dimethylbenz[a]anthracene. Progeny that were either homo- or heterozygous for the wild-type GH gene developed approximately 4 mammary tumors/rat, respectively. In contrast, SDR progeny developed only 0.21 tumors/rat. Mammary glands of SDRs had substantially less alveolar development compared with wild-type, yet ductal branching was similar in the two strains. Infusion of rat GH to SDRs induced mammary epithelial cell proliferation and alveolar development similar to that of wild-type rats. Taken together, these results demonstrate an important role for GH in alveolar development in the virgin rat, and provide the first direct evidence that GH plays a critical role in mammary carcinogenesis. PMID- 12082020 TI - Carnosol, an antioxidant in rosemary, suppresses inducible nitric oxide synthase through down-regulating nuclear factor-kappaB in mouse macrophages. AB - Carnosol is a naturally occurring phytopolyphenol found in rosemary. Carnosol functions as antioxidant and anticarcinogen. In the present study, we compared the antioxidant activity of carnosol and other compounds extracted from rosemary. Carnosol showed potent antioxidative activity in alpha,alpha-diphenyl-beta picrylhydrazyl (DPPH) free radicals scavenge and DNA protection from Fenton reaction. High concentrations of nitric oxide (NO) are produced by inducible NO synthase (iNOS) in inflammation and multiple stages of carcinogenesis. Treatment of mouse macrophage RAW 264.7 cell line with carnosol markly reduced lipopolysaccharide (LPS)-stimulated NO production in a concentration-related manner with an IC50 of 9.4 microM; but other tested compounds had slight effects. Western blot, reverse transcription-polymerase chain reaction, and northern blot analyses demonstrated that carnosol decreased LPS-induced iNOS mRNA and protein expression. Carnosol treatment showed reduction of nuclear factor-kappaB (NF kappaB) subunits translocation and NF-kappaB DNA binding activity in activated macrophages. Carnosol also showed inhibition of iNOS and NF-kappaB promoter activity in transient transfection assay. These activities were referred to down regulation of inhibitor kappaB (IkappaB) kinase (IKK) activity by carnosol (5 microM), thus inhibited LPS-induced phosphorylation as well as degradation of IkappaBalpha. Carnosol also inhibited LPS-induced p38 and p44/42 mitogen activated protein kinase (MAPK) activation at a higher concentration (20 microM). These results suggest that carnosol suppresses the NO production and iNOS gene expression by inhibiting NF-kappaB activation, and provide possible mechanisms for its anti-inflammatory and chemopreventive action. PMID- 12082021 TI - Inhibition of chronic ulcerative colitis-associated colorectal adenocarcinoma development in a murine model by N-acetylcysteine. AB - Long-term ulcerative colitis (UC) patients are at increased risk for developing colorectal cancer. In order to develop strategies for preventing UC-associated carcinogenesis, we studied the effect of the antioxidant N-acetylcysteine (NAC) on UC-associated cancer development in a mouse model. Female C57BL/6J mice were subjected to long-term administration of dextran sulfate sodium (DSS) in the drinking fluid and 2-fold iron-enriched AIN76A diet, with or without NAC. In the DSS-plus-2-fold iron positive control group, gross tumor incidence was 88.5% (23/26 mice) after 12 DSS cycles (1 DSS cycle = 7 day DSS treatment period followed by 10 day recovery period). The tumor multiplicity was 2.1 +/- 0.2 tumors/tumor-bearing mouse, and the tumor volume was 0.054 +/- 0.019 cm3. With 0.2% NAC administration, tumor incidence was significantly reduced (68%, 17/25 mice; P < 0.05), as was the tumor multiplicity (1.5 +/- 0.1 tumors/tumor-bearing mouse; P < 0.05). The tumor volume was lower (0.014 +/- 0.004 cm3), but not significantly decreased. The proliferation index was significantly decreased in non-cancerous epithelia (48.5 +/- 6.0% vs 32.0 +/- 3.7%; P < 0.05), but not in tumor cells. NAC significantly induced apoptosis in both non-cancerous epithelia and colorectal adenocarcinoma. The number of cells immunostained-positive for nitrotyrosine was markedly decreased in the non-cancerous mucosa of NAC-treated mice (102.4 +/-16.6 positive cells/mm2 mucosa vs 53.6 +/- 14.9 cells/mm2; P < 0.05). In addition, the number of inducible nitric oxide synthase (iNOS)-positive inflammatory cells in the non-cancerous mucosa of the distal colon was markedly decreased by NAC. This study indicates that the antioxidant NAC has the potential to serve as a preventive agent for UC-associated colorectal cancer, possibly via inhibition of cellular proliferation and nitrosative stress-caused cellular damage. PMID- 12082022 TI - Genetic polymorphisms of DNA repair and xenobiotic-metabolizing enzymes: role in mutagen sensitivity. AB - Mutagen sensitivity, measuring the extent of chromosome damage induced by an in vitro treatment of peripheral lymphocytes with bleomycin, has been associated with an increased risk of various human cancers. Sensitivity to bleomycin appears to have high heritability and is usually considered to reflect individual capacity to repair DNA lesions. Another potential contributor to variation in bleomycin sensitivity could be inherited differences in the metabolism of bleomycin. We assessed whether genetic polymorphisms of DNA repair and xenobiotic metabolizing enzymes (XMEs) could explain bleomycin sensitivity. Frequencies of bleomycin-induced chromatid breaks per cell (b/c) were determined for 80 healthy Caucasians. Genotypes of DNA repair genes XRCC (X-ray repair cross-complementing) 1 and 3 and XME genes bleomycin hydrolase (BLHX), glutathione S-transferase M1 (GSTM1) and T1 (GSTT1) and N-acetyltransferase 2 (NAT2) were analyzed from leukocyte DNA using methods based on polymerase chain reaction. The mean number of chromatid b/c was increased in individuals with XRCC1 codon 280 variant allele (P = 0.002; two-sided Mann-Whitney test). Smokers carrying BLHX codon 1450 variant allele showed a decrease in the mean number of chromatid b/c (P = 0.036). In multiple linear regression models including adjustment for age, sex, smoking and genotype, the adjusted relative risks (and 95% confidence intervals) were 1.18 (0.98-1.41) and 0.84 (0.69-1.00) for carriers of XRCC1 codon 280 and BLHX codon 1450 variant alleles, respectively. XRCC1 codon 280 polymorphism had a significant effect (P = 0.012) in predetermining whether the individual was classified as non-sensitive, sensitive or hypersensitive to bleomycin. Although based on relatively few individuals, our results suggest that bleomycin sensitivity is partially explained by genetic polymorphisms affecting DNA repair (XRCC1) and in vitro metabolism of bleomycin (BLHX). PMID- 12082023 TI - Metabolism of sinigrin (2-propenyl glucosinolate) by the human colonic microflora in a dynamic in vitro large-intestinal model. AB - Cruciferous vegetables, such as Brassica, which contain substantial quantities of glucosinolates, have been suggested to possess anticarcinogenic activity. Cutting and chewing of cruciferous vegetables releases the thioglucosidase enzyme myrosinase, which degrades glucosinolates to isothiocyanates and other minor metabolites. Cooking of cruciferous vegetables inactivates the myrosinase enzyme, allowing intact glucosinolates to reach the large intestine, where they can be degraded by the indigenous microflora into isothiocyanates. This local release of isothiocyanates may explain the protective effect of cruciferous vegetables on the colon epithelium. However, little is known about the amounts and identities of glucosinolate metabolites produced by the human microflora. The production of allyl isothiocyanate from sinigrin was investigated in a dynamic in vitro large intestinal model, after inoculation with a complex microflora of human origin. Sinigrin and allyl isothiocyanate concentrations were analysed in the lumen and dialysis fluid of the model. Peak levels of allyl isothiocyanate were observed between 9 and 12 h after the addition of sinigrin. The model was first set up with a pooled and cultured human microflora, in which 1 and 4% of, respectively, 1 and 15 mM sinigrin, was converted into AITC. However, the conversion rate was remarkably higher if different individual human microflora were used. Between 10% and 30% (mean 19%) of the sinigrin was converted into allyl isothiocyanate. The results of this study suggest that allyl isothiocyanate is converted further into other, yet unknown, metabolites. PMID- 12082024 TI - Inhibitor of apoptosis protein-1 promotes tumor cell survival in mesothelioma. AB - Malignant pleural mesothelioma (MPM) is a highly lethal pleural neoplasm that is often resistant to chemotherapeutic drugs, including cisplatin, and for which little is known regarding carcinogenic pathways. We used differential display to compare gene expression patterns in mesothelioma, normal pleura and normal lung, in order to better understand MPM pathobiology, and to search for genes that may facilitate drug resistance in this cancer. The human inhibitor of apoptosis protein-1 gene (IAP-1/MIHC/cIAP2) was discovered to be highly expressed in MPM. We confirmed overexpression of IAP-1 mRNA and protein in 39 additional human MPM tumor specimens and 3/5 (60%) MPM cell lines by multiple methods, including real time quantitative reverse transcription-PCR and western blot analysis. Using an antisense targeting approach, we found that attenuation of IAP-1 mRNA levels decreases baseline cell viability and increases the sensitivity of MPM cell lines to cisplatin by nearly 20-fold. Reduced IAP-1 gene expression also results in a concordant increase of the pro-apoptotic cleavage product of caspase 9 and a reduction in the number of viable tumor cells. Our observations strongly suggest that IAP-1 is at least partly responsible for promoting carcinogenesis and mediating resistance to cisplatin in many MPM tumors and that further study of this apoptotic pathway is warranted. PMID- 12082025 TI - The onset of p53-dependent DNA repair or apoptosis is determined by the level of accumulated damaged DNA. AB - The p53 tumor suppressor gene plays an important role in both apoptosis and DNA repair pathways that are pivotal for genomic stability. Here we show that the treatment of cells with low doses of gamma-irradiation or cisplatin resulted in an immediate enhancement of p53-dependent DNA repair, measured by base excision repair (BER) activity. However, treatment of cells with high doses of DNA damaging agents resulted in a reduction in p53-dependent DNA repair and in the induction of p53-dependent apoptosis. Analysis of p53 upstream molecular events suggested that regulation of p53-associated DNA repair is ATM-dependent. Furthermore, we observed that while dephosphorylation of Ser376 at the C-terminus of the p53 protein was associated with enhancement in DNA repair, phosphorylation at the N-terminal Ser15 resulted in the reduction in DNA repair. The latter is also in correlation with an enhancement in the specific DNA binding activity and in the induction of apoptosis. Treatment of cells with a caspase inhibitor, prior to the damaging agent-blocked apoptosis, had no effect on the DNA repair pattern. Taken together, this suggests that the decision of cells to induce a p53 dependent DNA repair or apoptosis is most probably controlled by the level of genotoxic agent introduced to cells. PMID- 12082026 TI - Loss of heterozygosity in malignant rat schwannomas chemically induced in hybrids of inbred rat strains with differential tumor susceptibility. AB - Rats of the inbred BD strains strongly differ in their susceptibility to the induction of tumors of the central (CNS) and peripheral nervous system (PNS) by N ethyl-N-nitrosourea (EtNU). Malignant schwannomas induced in (BDIX x BDIV) and (BDIX x BDVI) rat hybrids were analyzed to identify genetic alterations associated with EtNU-induced tumorigenesis in the PNS. EtNU-induced schwannomas exclusively exhibit an A:T T:A transversion mutation of the neu/Erbb-2 gene located on chromosome 10, with subsequent loss of the wild-type neu/Erbb-2 allele at a post-initiation stage. Targeted allelic deletion mapping previously revealed losses of heterozygosity (LOH) at the distal end of chromosome 10 in a large majority of (BDIX x BDIV) schwannomas. The aims of the present study were (i) to scan the whole genome for further LOHs; (ii) to narrow down the consensus regions of frequently occurring allelic deletions using tumors from different crosses of BD rats; and (iii) to determine the sequence of genetic alterations during schwannoma development. A limited number of (BDIX x BDIV) F(1) tumors were initially screened for LOH and microsatellite instability (MI) by amplifying 58 microsatellite markers spanning the whole genome. LOHs on chromosome 5 were detected in 9/17 tumors, with random loss of the parental alleles. Ninety-two schwannomas from different BD rat-crosses were then analyzed to solidify these data and to determine the consensus region of frequent LOHs. The results indicate that LOHs on chromosomes 10 and 5 are required for the development of EtNU induced malignant schwannomas from immature neu/Erbb-2 mutant glial cells, and that putative tumor suppressor genes are localized on chromosome 10q32.3, corresponding to human chromosome 17q25.3, and the telomeric region of mouse chromosome 11, and on the telomeric quarter of chromosome 5. MI was detected in <0.2% of cases. PMID- 12082028 TI - Molecular profiling of genes up-regulated during promotion by phenobarbital treatment in a medium-term rat liver bioassay. AB - In search of genes that are steadily up-regulated during the promotion stage in carcinogenesis, suppression PCR subtractive hybridization and following northern blot screening were performed using a phenobarbital (PB)-promotion model based on a medium-term liver bioassay. Two weeks after a single injection of diethylnitrosamine (DEN; 200 mg/kg body wt, i.p.), rats were given 600 p.p.m. PB in the drinking water for up to 64 weeks. For comparison, animals fed 1 p.p.m. ethinylestradiol (EE) or 3000 p.p.m. butylated hydroxytoluene (BHT) in the diet at promotion stage were also included. Rats were subjected to partial hepatectomy (PH) at week 3. In addition, dose-dependence of PB at week 8 of promotion and responsiveness to representative non-genotoxic carcinogens without DEN initiation were examined. Fragments of a total of 67 different genes were isolated from the up-regulated gene population in the liver at day 10 of PB treatment by subtracting from basal expression of DEN + PH alone. Using northern blot screening for signal-detectable 48 genes, 16 genes showed up-regulation in the livers at week 8 of promotion, common to the PB and EE treatments with the levels being three times or more than the basal expression of unpromoted liver. The majority of these genes were also up-regulated at week 8 by BHT treatment, and were also constitutively expressed in the DEN(-), PH(-) untreated rat livers. Among the up-regulated genes common to the PB and EE promotion, and not responding to the non-genotoxic carcinogens in uninitiated liver, the following six genes showed overexpression in PB-promoted hepatocellular carcinomas at week 64, with the levels three times or more than untreated rat liver: ubiquitously expressed mammalian ABC half transporter, apolipoprotein A4, nuclear receptor binding factor-2, CD81, hypothetical protein (HSPC014) and one unidentified gene. These genes might be candidates for biomarkers in screening of non-genotoxic hepatocarcinogens by analysis in two-stage carcinogenesis models. PMID- 12082027 TI - Low amounts of the DNA repair XPA protein are sufficient to recover UV resistance. AB - DNA integrity is threatened by the damaging effects of physical and chemical agents that can affect its function. Nucleotide excision repair (NER) is one of the most known and flexible mechanisms of DNA repair. This mechanism can recognize and remove damages causing DNA double-helix distortion, including the cyclobutane pyrimidine dimers (CPDs) and the pyrimidine-pyrimidone (6-4) photoproducts, promoted by ultraviolet light (UV). The human syndrome xeroderma pigmentosum (XP) is clinically characterized chiefly by the early onset of severe photosensitivity of the exposed regions of the skin, a very high incidence of skin cancers and frequent neurological abnormalities. The xpa gene seems to be involved during UV damage recognition, in both global genome repair (GGR) and transcription-coupled repair (TCR). The modulation of xpa expression may modify the DNA repair rate in the cell genome, providing a valuable contribution to an understanding of the NER process. The controlled expression of the cDNA xpa in XP12RO deficient cells was achieved through the transfection of a muristerone-A inducible vector, pINXA. The INXA15 clone shows good induction of the XPA protein and total complementation of XP12RO cell deficiency. Overexpression of this protein resulted in UV cell survival comparable to normal control human cells. Moreover, low expression of the XPA protein in these cells is sufficient for total complementation in cellular UV sensitivity and DNA repair activity. These data demonstrate that XPA protein concentration is not a limiting factor for DNA repair. PMID- 12082029 TI - Single copy heterozygote integration of HPV 33 in chromosomal band 5p14 is found in an epithelial cell clone with selective growth advantage. AB - Infection with human papillomavirus (HPV) of specific high-risk type triggers a series of events in target cells, which will eventually lead to development of genital neoplasia. The integration of high-risk HPV DNA into the cell genome has been regarded as a crucial event in tumor progression. With respect to different HPV types, the knowledge of HPV integrated loci is still limited. We have now determined the genomic variation and chromosomal location of HPV 33 DNA in the cell line UT-DEC-1, established from a vaginal mild dysplasia lesion. The viral sequence of the cell line was determined, and a variant of the prototype HPV 33 strain was identified, showing nucleotide substitutions resulting in amino acid changes in the E2, L2 and E4 open reading frames. In late passage UT-DEC-1 cells, a deletion of more than half of the 3' part of E1 and major parts of the E2 and E4 genes provided evidence for integration. The flanking sequences of the integration site were completely homologous to published sequences from chromosomal band 5p14, and remained unchanged in all subclones established from late passage cells. There were no chromosomal deletions or gross rearrangements at the integration site, and only a single heterozygotic copy of HPV 33 was detected. The karyotype of late passage cells showed only minor changes compared with early passage cells. During passaging of the cell line, there were progressive changes towards a malignant phenotype, and in parallel to this, the cells carrying episomal HPV 33 of the early passages was completely superseded by cells containing the integrated virus. Thus, our results show that this single copy heterozygote integration of HPV 33 into chromosome band 5p14 appears to be associated with emergence of cells escaping senescence, and with growth advantage compared with cells carrying episomal virus. PMID- 12082030 TI - Membrane-type matrix metalloproteinases mediate curcumin-induced cell migration in non-tumorigenic colon epithelial cells differing in Apc genotype. AB - Colonic epithelial cell migration is required for normal differentiated cell function. This migratory phenotype is dependent upon wild-type adenomatous polyposis coli (Apc) expression. Non-tumorigenic murine colon epithelial cell lines with distinct Apc genotypes, i.e. young adult mouse colon (YAMC; Apc(+/+)) and immortomouse/Min colon epithelial (IMCE; Apc(Min/+) cells) were used to assess the association between the Apc genotype, cell motility and matrix metalloproteinase (MMP) activity. Cells were treated with epidermal growth factor (EGF; 1, 10 and 25 ng/ml), hepatocyte growth factor (HGF; 1, 10 and 25 ng/ml) and/or curcumin (0.1-100 microM). EGF (25 ng/ml) and HGF (25 ng/ml) induced a greater migratory response in YAMC compared with IMCE cells after 24 h (P < 0.05). Treatment with curcumin induced a greater or equivalent migratory response in IMCE than YAMC cells. When migrating cells were treated with Ilomastat (MMP inhibitor), migration was inhibited in both cell types. High concentrations of Ilomastat (25 and 50 microM) inhibited migration in both cell types, while low concentrations (10 microM) inhibited HGF-induced IMCE migration. Curcumin-induced migration was inhibited in both cell types at the highest concentration of Ilomastat (50 microM). Immuno-localization analysis of membrane type-1 (MT1)-MMP indicated that migration is associated with the redistribution of this protein from the endoplasmic reticulum to the plasma membrane. Addition of neutralizing polyclonal antibodies against MT1-MMP or a mixture of MT1, 2- and 3-MMPs demonstrated partial or complete inhibition of cell migration in both cell types, respectively. The data provide the first evidence that migration in non tumorigenic murine colon epithelial cells is: (i) inducible by EGF and HGF in an Apc genotype-dependent manner, (ii) dependent on MT-MMP activity and (iii) inducible by curcumin in an Apc genotype-independent manner. The data suggest a potential mechanism by which curcumin may induce cells heterozygous for Apc to overcome defective cell migration, a phenotype associated with cell differentiation and apoptosis. PMID- 12082031 TI - Catechol ortho-quinones: the electrophilic compounds that form depurinating DNA adducts and could initiate cancer and other diseases. AB - Catechol estrogens and catecholamines are metabolized to quinones, and the metabolite catechol (1,2-dihydroxybenzene) of the leukemogenic benzene can also be oxidized to its quinone. We report here that quinones obtained by enzymatic oxidation of catechol and dopamine with horseradish peroxidase, tyrosinase or phenobarbital-induced rat liver microsomes react with DNA by 1,4-Michael addition to form predominantly depurinating adducts at the N-7 of guanine and the N-3 of adenine. These adducts are analogous to the ones formed with DNA by enzymatically oxidized 4-catechol estrogens (Cavalieri,E.L., et al. (1997) PROC: Natl Acad. Sci., 94, 10937). The adducts were identified by comparison with standard adducts synthesized by reaction of catechol quinone or dopamine quinone with deoxyguanosine or adenine. We hypothesize that mutations induced by apurinic sites, generated by the depurinating adducts, may initiate cancer by benzene and estrogens, and some neurodegenerative diseases (e.g. Parkinson's disease) by dopamine. These data suggest that there is a unifying molecular mechanism, namely, formation of specific depurinating DNA adducts at the N-7 of guanine and N-3 of adenine, that could initiate many cancers and neurodegenerative diseases. PMID- 12082032 TI - Myeloid, B and T lymphoid and mixed lineage thymic lymphomas in the irradiated mouse. AB - Thymic lymphoma is a very common spontaneous and/or induced malignancy in both inbred mice and in transgenic mouse models of human cancer. Although a thymic lymphoma is defined as thymus-dependent T-cell malignancy, diagnostic criteria vary between studies and considerable heterogeneity has been reported. To define and classify the thymic lymphomas that arose in our study of X-irradiated (CBA/HxC57BL/6)F1, F1 backcross and F1 intercross mice, 66 thymic lymphomas were immunogenotyped for immunoglobulin heavy chain (IgH) and T-cell receptor beta (TCRbeta) gene rearrangements, and/or analysed for expression of lineage-specific markers and allelic loss on chromosome 4. The data indicate that 33% of the thymic lymphomas are very similar to mouse radiation-induced acute myeloid (AML) and mixed lineage (IgH(R), TCRbeta(G)) pre-B lympho-myeloid (L-MLs) leukaemias, 33% are mixed lineage (IgH(R), TCRbeta(R)) B/T lymphoid and <33% can be described as single lineage (IgH(G), TCRbeta(R)) T-cell malignancies. As the myeloid and L ML leukaemias are not thymus-dependent this suggests that a malignant myeloid or pre-B lympho-myeloid cell can colonize the spleen to give an AML or L-ML leukaemia, or can colonize the thymus where TCRbeta gene rearrangement(s) may be induced to give the mixed lineage thymic lymphomas. Thus, assuming the single lineage T-cell thymic lymphomas fulfil the criteria of a thymus-dependent T-cell malignancy, thymic lymphomas are comprised of at least three distinct malignancies. PMID- 12082033 TI - Cancer, genes and gender. PMID- 12082035 TI - Head drop and camptocormia. PMID- 12082036 TI - "Normal pressure hydrocephalus": what's in a name? PMID- 12082038 TI - Minor head injury as cause and co-factor in the aetiology of stroke in childhood: a report of eight cases. AB - BACKGROUND: Traumatic stroke usually occurs after dissection of large extracranial or intracranial vessels, leading to disseminated cerebral embolism. Stretching and distorting forces in cerebral intraparenchymal end arteries can cause intimal lesions followed by an occluding thrombus. OBJECTIVE: To investigate the importance of traumatic endothelial lesions in intraparenchymal end arteries after minor head injuries. METHODS: The cases of eight children are reported. They were aged between two and seven years (mean 6.2 years), and they developed significant neurological deficits at 15 minutes to 72 hours (mean 16.3 hours) after minor head injuries. RESULTS: The the patients all had hemiparesis combined with other signs, including central facial paralysis, dysphasia, dysphagia, and extrapyramidal signs. Computed tomography or magnetic resonance imaging showed cerebral infarctions affecting branches of the middle cerebral artery (n = 3), anterior cerebral artery (n = 1), posterior cerebral artery (n = 1), and basilar artery (n = 3). These lesions affected the basal ganglia, the internal capsule, and the brain stem. Neither heart disease nor dissections of large vessels were present. Two children had prothrombotic risk factors (an increase in lipoprotein (a) and a factor V Leiden mutation). The follow up period was between three months and 13 years (mean 3.9 years). Outcome was classified according to the Glasgow outcome scale as moderate disability (n = 4), severe disability (n = 2), non-disabling sequelae (n = 1), and total recovery (n = 1). CONCLUSIONS: Minor head injuries can be cause and co-factor in the aetiology of stroke. The frequency of this may be underestimated, and detailed medical history of the days before stroke manifestation may identify more traumatic events, especially in the group of so called "idiopathic" strokes. PMID- 12082037 TI - Is normal pressure hydrocephalus a valid concept in 2002? A reappraisal in five questions and proposal for a new designation of the syndrome as "chronic hydrocephalus". AB - The authors question the current validity of the traditional concept of normal pressure hydrocephalus (NPH) as it was described by Adams and Hakim in 1965. The classic features of the disease are addressed. It is concluded that most of the historical statements made three decades ago need to be revised. Especially, the term "normal pressure" hydrocephalus probably does not match the actual manometric profile of patients with NPH. Similarly, the terms"curable"and "reversible" dementia are inadequate to designate the mental alterations of NPH. That NPH is non-specific to the adult population is also stressed, since it may be not uncommonly encountered in paediatrics, especially in an implanted shunt malfunction. The term "chronic hydrocephalus" without reference to cerebrospinal fluid pressure and to the age of the patient is proposed instead of NPH, which seems out of step with current knowledge of the pressure profile and with the diagnosis and decision making context in patients with so called NPH. PMID- 12082039 TI - Feasibility and validity of transcranial duplex sonography in patients with acute stroke. AB - OBJECTIVES: To evaluate in a prospective multicentre setting the feasibility of transcranial colour coded duplex sonography (TCCS) for examination of the middle cerebral artery (MCA) in patients with acute hemispheric stroke, and to assess the validity of sonographic findings in a subgroup of patients who also had a correlative angiographic examination. METHODS: TCCS was performed in 58 consecutive patients within six hours of the onset of a moderate to severe hemispheric stroke. Ultrasound contrast agent (Levovist) was applied if necessary. Thirty two patients also had computed tomography angiography (n=13), magnetic resonance angiography (n=18), or digital subtraction angiography (n=1). In 14 of these patients, both the sonographic and corresponding angiographic examination were performed within six hours of stroke onset (mean time difference between TCCS and angiography 0.8 hours). Eighteen patients, in whom angiography was carried out more than 24 hours after stroke onset, had a follow up TCCS for method comparison (mean time difference 6.1 hours). RESULTS: Initial unenhanced TCCS performed 3.4 (SD 1.2) hours after the onset of symptoms depicted the symptomatic MCA mainstem in 32 patients (55%) (13 occlusions, one stenosis, 18 patent arteries). After signal enhancement, MCA status could be determined in 54 patients (93%) (p<0.05), showing an occlusion in 25, a stenosis in two, and a patent artery in 27 patients. In 31 of the 32 patients who had correlative angiography, TCCS and angiography produced the same diagnosis of the symptomatic MCA (10 occlusions, three stenoses, 18 patent arteries); TCCS was inconclusive in the remaining one. CONCLUSION: TCCS is a feasible, fast, and valid non-invasive bedside method for evaluating the MCA in an acute stroke setting, particularly when contrast enhancement is applied. It may be a valuable and cost effective alternative to computed tomography and magnetic resonance angiography in future stroke trials. PMID- 12082040 TI - Outcome, observer reliability, and patient preferences if CTA, MRA, or Doppler ultrasound were used, individually or together, instead of digital subtraction angiography before carotid endarterectomy. AB - OBJECTIVES: To evaluate the accuracy of routinely available non-invasive tests (spiral computed tomographic angiography (CTA), time of flight magnetic resonance angiography (MRA), and colour Doppler ultrasound (DUS)), individually and together, compared with intra-arterial digital subtraction angiography (DSA) in patients with symptomatic tight carotid stenosis; and to assess the effect of substituting non-invasive tests for DSA on outcome, interobserver variability, and patient preference. METHODS: Patients referred from a neurovascular clinic were subjected prospectively to DUS imaging. The operator was blind to symptoms. Patients with a tight carotid stenosis on the symptomatic side were admitted for DSA. CTA and MRA were performed during the admission. The CTA, MRA, and DSA films were each read independently by two of six experienced radiologists, blind to all other data. RESULTS: 67 patients were included (34 had all four imaging procedures). DUS, CTA, and MRA all agreed with DSA in the diagnosis of operable v non-operable disease in about 80% of patients. CTA tended to underestimate (sensitivity 0.65, specificity 1.0), MRA to overestimate (sensitivity 1.0, specificity 0.57), and DUS to agree most closely with (sensitivity 0.85, specificity 0.71) the degree of stenosis as shown by DSA. When using any two of the three non-invasive tests in combination, adding the third if the first two disagreed would result in very few misdiagnoses (about 6%). MRA had similar interobserver variability to CTA (both worse than DSA). Patients preferred CTA over MRA and DSA. CONCLUSIONS: DUS, CTA, and MRA all show similar accuracy in the diagnosis of symptomatic carotid stenosis. No technique on its own is accurate enough to replace DSA. Two non-invasive techniques in combination, and adding a third if the first two disagree, appears more accurate, but may still result in diagnostic errors. PMID- 12082041 TI - Transoesophageal echocardiography in selecting patients for anticoagulation after ischaemic stroke or transient ischaemic attack. AB - OBJECTIVES: To investigate prospectively the role of transoesophageal echocardiography (TEE) in selecting patients for anticoagulation in an unselected stroke population. METHODS: Transthoracic echocardiography (TTE) and TEE were done in all clinically suitable hospitalised patients (n = 457) with transient ischaemic attack or ischaemic stroke in the acute phase during a two year period in Turku University Hospital. 441 patients were successfully evaluated for cardiac sources of embolism using TEE within 31 days of the event. RESULTS: A major risk factor for a cardiac source of embolism excluding atrial fibrillation, acute myocardial infarction, and prosthetic valve was detected in 10% of patients and a minor risk factor for a cardiac source of embolism in 46%. When a major risk factor of a cardiac source of embolism was detected using TTE or TEE and no contraindications were present, the patient was given anticoagulation drugs. If a minor risk factor for a cardiac source of embolism was detected, anticoagulation treatment was started after clinical assessment, if no contraindications were present. In 62 (14%) cases, the patient was given oral anticoagulation drugs or the necessity of ongoing anticoagulation treatment was confirmed on the basis of TEE. When these anticoagulation treated patients were evaluated using logistic regression analysis, they were found to have significantly more atrial fibrillation and histories of myocardial infarctions. Moreover, the patients were mainly men. When patients in sinus rhythm and without any history of cardiac disease were analysed, 8% of patients were found to have been given anticoagulation drugs on the basis of TEE data. CONCLUSION: This study suggests that TEE should be used in patients with stroke even without any clinical evidence of cardiac disease when the patients are candidates for anticoagulation. PMID- 12082042 TI - Cerebral haemodynamics and depression in the elderly. AB - BACKGROUND: Evidence from epidemiological and neuroimaging studies suggests that cerebrovascular disease is associated with depressive disorders in the elderly, but the extent to which it contributes to the pathogenesis of late life depression is unclear. OBJECTIVE: To investigate the relation between cerebral haemodynamics and depression in a population based study, using transcranial Doppler ultrasonography. METHODS: Cerebral blood flow velocity and CO2 induced vasomotor reactivity in the middle cerebral artery were measured in 2093 men and women who participated in the Rotterdam study. All subjects were screened for depressive symptoms using the Center of Epidemiological Studies Depression scale, and those with a score of 16 or over had a psychiatric work up. In a semistructured interview, diagnoses of depressive disorders according to the DSM IV and subthreshold depressive disorder were established. Analyses of covariance controlled for age, sex, stroke, cognitive score, and cardiovascular risk factors were used to compare means of haemodynamic variables. RESULTS: Subjects with depressive symptoms had reduced blood flow velocities (mean difference, -2.9 cm/s; 95% confidence interval (CI), -5.0 to -0.8; p = 0.008) and lower vasomotor reactivity (mean difference -0.5%/kPa; 95% CI, -1.0 to -0.05; p = 0.03). Blood flow velocity was reduced most in subjects suffering from a DSM-IV depressive disorder (mean difference, -4.9 cm/s; 95% CI, -8.5 to -1.4; p = 0.006). The overall reduction in vasomotor reactivity was accounted for by subjects with subthreshold depressive disorder. CONCLUSIONS: Depression in late life is associated with cerebral haemodynamic changes that can be assessed by transcranial Doppler ultrasonography. The observed reduction in cerebral blood flow velocity could be a result of reduced demand in more seriously depressed cases with a DSM-IV disorder, whereas reduced CO2 induced cerebral vasomotor reactivity is a possible causal factor for subthreshold depressive disorder. PMID- 12082043 TI - Enduring increased risk of developing depression and mania in patients with dementia. AB - OBJECTIVE: To investigate the time relation between dementia and major affective disorders (major depression and mania). METHODS: Register linkage study of the Danish Hospital Register and the Danish Psychiatric Central Research Register, to establish study cohorts of patients with dementia and control groups (osteoarthritis or diabetes) on first discharge from hospital. Follow up of cohorts was for up to 21 years. Hazard of death was allowed for by the use of competing risks models. RESULTS: Patients with dementia had an increased risk of being admitted to hospital for major depression or mania during the course of the illness. The incidence remained elevated throughout the rest of the patient's life. CONCLUSIONS: Patients with dementia have an increased risk of developing depression or mania. Proper treatment of affective disorders in patients with dementia is important in reducing suffering and costs. PMID- 12082044 TI - Inflammation and neuropathic attacks in hereditary brachial plexus neuropathy. AB - OBJECTIVE: To study the role of mechanical, infectious, and inflammatory factors inducing neuropathic attacks in hereditary brachial plexus neuropathy (HBPN), an autosomal dominant disorder characterised by attacks of pain and weakness, atrophy, and sensory alterations of the shoulder girdle and upper limb muscles. METHODS: Four patients from separate kindreds with HBPN were evaluated. Upper extremity nerve biopsies were obtained during attacks from a person of each kindred. In situ hybridisation for common viruses in nerve tissue and genetic testing for a hereditary tendency to pressure palsies (HNPP; tomaculous neuropathy) were undertaken. Two patients treated with intravenous methyl prednisolone had serial clinical and electrophysiological examinations. One patient was followed prospectively through pregnancy and during the development of a stereotypic attack after elective caesarean delivery. RESULTS: Upper extremity nerve biopsies in two patients showed prominent perivascular inflammatory infiltrates with vessel wall disruption. Nerve in situ hybridisation for viruses was negative. There were no tomaculous nerve changes. In two patients intravenous methyl prednisolone ameliorated symptoms (largely pain), but with tapering of steroid dose, signs and symptoms worsened. Elective caesarean delivery did not prevent a typical postpartum attack. CONCLUSIONS: Inflammation, probably immune, appears pathogenic for some if not all attacks of HBPN. Immune modulation may be useful in preventing or reducing the neuropathic attacks, although controlled trials are needed to establish efficacy, as correction of the mutant gene is still not possible. The genes involved in immune regulation may be candidates for causing HBPN disorders. PMID- 12082046 TI - Neuropsychological and quantitative oculometric study of a case of sporadic Creutzfeldt-Jakob disease at predementia stage. AB - A quantitative assessment of eye movements and a detailed neuropsychological profile were conducted at predementia stage in a patient who later had histological confirmation of sporadic Creutzfeldt-Jakob disease (CJD). The patient was a middle aged man who presented with abnormal eye movements and poor balance. Neuropsychological deficits suggested orbito-mesial dysfunction, resembling progressive supranuclear palsy. Oculometry showed accurate but dramatically slowed saccades, with normal pursuit movements. Neuropsychology and quantitative oculometry may be of value in the differential diagnosis and earlier detection of dementia-akinetic-rigid syndromes; in particular, because of the highly stereotyped nature of saccades, routine quantitative oculometry can reveal significant impairment at a very early stage in some cases and could thus facilitate earlier diagnosis. PMID- 12082045 TI - Quantitative oculographic characterisation of internuclear ophthalmoparesis in multiple sclerosis: the versional dysconjugacy index Z score. AB - BACKGROUND: There is a poor correlation between multiple sclerosis disease activity, as measured by magnetic resonance imaging, and clinical disability. OBJECTIVE: To establish oculographic criteria for the diagnosis and severity of internuclear ophthalmoparesis (INO), so that future studies can link the severity of ocular dysconjugacy with neuroradiological abnormalities within the dorsomedial brain stem tegmentum. METHODS: The study involved 58 patients with multiple sclerosis and chronic INO and 40 normal subjects. Two dimensional infrared oculography was used to derive the versional dysconjugacy index (VDI) the ratio of abducting to adducting eye movements for peak velocity and acceleration. Diagnostic criteria for the diagnosis and severity of INO were derived using a Z score and histogram analysis, which allowed comparisons of the VDI from multiple sclerosis patients and from a control population. RESULTS: For a given saccade, the VDI was typically higher for acceleration v velocity, whereas the Z scores for velocity measures were always higher than values derived from comparable acceleration VDI measures; this was related to the greater variability of acceleration measures. Thus velocity was a more reliable measure from which to determine Z scores and thereby the criteria for INO and its level of severity. The mean (SD) value of the VDI velocity derived from 40 control subjects was 0.922 (0.072). The highest VDI for velocity from a normal control subject was 1.09, which was 2.33 SD above the normal control mean VDI. We therefore chose 2 SD beyond this value (that is, a Z score of 4.33) as the minimum criterion for the oculographic confirmation of INO. Of patients thought to have unilateral INO on clinical grounds, 70% (16/23) were found to have bilateral INO on oculographic assessment. CONCLUSIONS: INO can be confirmed and characterised by level of severity using Z score analysis of quantitative oculography. Such assessments may be useful for linking the level of severity of a specific clinical disability with neuroradiological measures of brain tissue pathology in multiple sclerosis. PMID- 12082047 TI - Morphometry of the corpus callosum in patients with questionable and mild dementia. AB - Previous studies have shown a reduction in corpus callosum area in advanced Alzheimer's disease, but it is unclear whether callosal atrophy is present in the transitional phase between health and the onset of dementia. The aim of this study was to investigate whether callosal atrophy is present in subjects with questionable and mild dementia and to assess the diagnostic value of callosal measures. In 83 subjects aged 72 to 85 years (33 normal controls, 27 patients with questionable dementia, 23 with mild Alzheimer's disease), magnetic resonance images were recorded and the mid-sagittal callosal area measured. Significant differences were found between normal controls and mild dementia. In subjects with questionable dementia callosal size was intermediate between normal controls and mild Alzheimer's disease. However, callosal measures were unsuitable for diagnostic differentiation between healthy subjects, subjects with questionable dementia, and subjects with mild Alzheimer's disease. The severity of white matter changes did not differ between the groups. PMID- 12082048 TI - Association study of three polymorphisms of TGF-beta1 gene with Alzheimer's disease. AB - BACKGROUND: There is evidence that inflammatory processes may contribute to the development of Alzheimer's disease through production of cytokines and free radicals that damage neurones. A recent study has shown that transforming growth factor beta1 (TGF-beta1) signalling in astrocytes promotes Abeta production and could play a critical role in the formation of amyloid plaques in the brain. OBJECTIVES: To explore the impact of the -800 and -509 TGF-beta1 promoter polymorphisms and the +25 polymorphism on the risk of occurrence of Alzheimer's disease in a large population of sporadic cases and controls, and on the amyloid beta (Abeta) load in the brains of Alzheimer patients. METHODS: The TGF-beta1 genotypes of the three polymorphisms were determined in 678 sporadic Alzheimer's disease patients and 667 controls. They were also characterised, along with Abeta load, in the brains of 81 necropsy confirmed Alzheimer patients. RESULTS: No significant variations in the distribution of the genotypes and haplotypes were observed between Alzheimer patients and controls, or in the amount of Abeta deposition. CONCLUSIONS: These results do not suggest an influence of genetic variability at the TGF-beta1 gene locus on the occurrence of Alzheimer's disease. PMID- 12082049 TI - Familial isolated hyperCKaemia associated with a new mutation in the caveolin-3 (CAV-3) gene. AB - An 18 year old man and his mother both presented with persistent, isolated raised serum creatine kinase (hyperCKaemia) without muscle symptoms. Analysis of caveolin-3 protein expression in muscle biopsy of the propositus showed a reduction in the protein. Genetic analysis revealed a new heterozygous mutation in the caveolin-3 (CAV-3) gene: a C-->T transition at nucleotide position 83 in exon 1 leading to a substitution of a proline for a leucine at amino acid position 28 (P28L). This is the first pathogenic mutation in the CAV-3 gene associated with isolated familial hyperCKaemia. It expands the genetic heterogeneity in patients with caveolin-3 deficiency and confirms that caveolin-3 deficiency should be considered in the differential diagnosis of isolated hyperCKaemia. PMID- 12082050 TI - Progressive sudomotor dysfunction in amyotrophic lateral sclerosis. AB - Autonomic dysregulation is part of the complex degenerative process in amyotrophic lateral sclerosis (ALS). To investigate this, sweating was examined at rest in 39 patients with ALS in comparison with a control group. Sweat was collected over a 30 second period over the thenar and hypothenar eminences and on the sole of the foot, using a commercial device based on vapour pressure gradient. The measurements were repeated after three and six months in 10 patients for longitudinal analysis. In early ALS, patients had significantly higher skin water loss than control subjects over the thenar and the hypothenar eminences. In advanced disease stages, sweating was decreased at all sites compared with controls. A significant decline in sweat secretion of about 40% was found over a six month period. The findings suggest an abnormal sympathetic activity with hyperhidrosis in early ALS and a reduction in sweat production as the disease progresses. PMID- 12082051 TI - High blood pressure and decreased heart rate variability in the Cuban epidemic neuropathy. AB - Autonomic cardiovascular reflexes were investigated in patients with an epidemic optic and peripheral neuropathy, which affected more than 50 000 people in Cuba between 1991 and 1994 and was probably caused by nutritional deficiency. Affected patients had significantly higher blood pressure than age matched controls, both while supine and standing, and significantly lower heart rate variability during paced breathing, suggesting reduced cardiac parasympathetic innervation. PMID- 12082052 TI - Sex differences and lateral asymmetry in heart rate modulation in patients with temporal lobe epilepsy. AB - This study was designed to study the influence of gender and lateral hemispheric asymmetry on heart rate modulation during temporal lobe seizures. Heart rate was recorded during complex partial seizures in 10 female and 11 male patients (12 with a right temporal lobe focus, nine with a left focus), with simultaneous video monitoring and bilateral subdural electrode placement. Heart rate changes were analysed during the seizure, at a time when the epileptic activity was restricted to one hemisphere. In this analysed interval, the heart rate of patients with a right temporal lobe focus showed a significant increase, from 73.5 to 91.0 beats/min (F = 10.7, df = 2.3/27, p < 0.001), while the heart rate of patients with a left sided focus only increased slightly, from 77.0 to 82.5 beats/min (NS). An effect of sex was demonstrated, in that the influence of focus laterality could only be shown in male patients (F = 14.24, df = 2.58/27, p < 0.001). These results confirm the right hemispheric lateralisation of sympathetic cardiac control in male patients. PMID- 12082053 TI - An antibody to VacA of Helicobacter pylori in cerebrospinal fluid from patients with Guillain-Barre syndrome. AB - OBJECTIVE: To detect antibodies to recombinant vacuolating cytotoxin (r-VacA) of Helicobacter pylori in cerebrospinal fluid (CSF) from patients with Guillain Barre syndrome (GBS). METHODS: CSF samples from 13 patients with GBS (electrophysiologically classified as eight acute inflammatory demyelinating polyradiculoneuropathy (AIDP), four acute motor axonal neuropathy (AMAN), and one unexcitable nerve conduction) and eight disease control patients were studied. The r-VacA protein was separated by SDS/PAGE, and Western blot analysis was carried out. RESULTS: Six of the 13 patients with GBS had a specific IgG antibody to VacA of H pylori, which was confirmed by absorption experiments using r-VacA. Every patient with positive CSF anti-r-VacA IgG had AIDP. CONCLUSION: The sequence homology previously found between VacA and human (Na(+)+K(+))-ATPase A subunit suggests that antibodies to VacA involve ion channels in abaxonal Schwann cell plasmalemma resulting in demyelination in some patients with GBS. PMID- 12082054 TI - Phosphorylated tau in cerebrospinal fluid as a marker for Creutzfeldt-Jakob disease. AB - OBJECTIVE: To determine the concentrations of microtubule associated protein tau and multiple phosphorylated tau epitopes in the cerebrospinal fluid of patients with sporadic Creutzfeldt-Jakob disease (sCJD), dementias, and controls, in order to evaluate their diagnostic use and clinical relevance. METHODS: The CSF concentrations of total tau and phosphorylated tau at epitope 181 were determined by enzyme linked immunosorbent assay in 66 definite and nine probable sCJD patients, and in 97 controls. Other phosphorylated tau epitopes were investigated by western blot. RESULTS: In the sCJD population, determination of 14-3-3 protein and total tau protein concentrations was of the highest diagnostic value, with a sensitivity of 96% and 92%, respectively, and a specificity of 94% and 97%. Two distinct subgroups could be identified among the 75 sCJD patients based on the detection of phosphorylated tau at threonine 181 and serines 199, 202, and 404. A high phosphorylated tau concentration was clinically correlated with a significantly shorter disease duration, early onset of akinetic mutism, and a higher rate of typical EEGs (p < 0.05). CONCLUSIONS: Although the determination of phosphorylated tau levels cannot be used as a diagnostic biomarker, it may prove useful for estimating the prognosis of an sCJD patient. These experiments reconfirm that sCJD is a disease with a complex pathology. PMID- 12082055 TI - Constantin von Economo (1876-1931). PMID- 12082056 TI - Louis Pasteur and rabies: a brief note. PMID- 12082057 TI - Don't throw in the towel! A case of reversible coma. AB - A young woman with pre-eclampsia became unresponsive shortly after delivery. Examination revealed extensive brain stem dysfunction with absent pupillary light reflexes and decerebrate posturing. Computed tomography showed hypodensity throughout the brain stem, and it was initially thought that she had suffered catastrophic brain stem infarction. However, magnetic resonance diffusion imaging and apparent diffusion coefficient mapping showed that she had brain stem vasogenic oedema (posterior reversible encephalopathy syndrome, PRES), rather than cytotoxic oedema. With antihypertensive and supportive treatment, she recovered rapidly, and had no abnormalities on repeat imaging. PMID- 12082058 TI - Tumefactive multiple sclerosis plaque. PMID- 12082059 TI - "Sporadic" familial amyloidotic polyneuropathy in a German patient with B cell lymphocytic leukaemia. PMID- 12082060 TI - Aneurysm of the extracranial internal carotid artery presenting as the syndrome of glossopharyngeal pain and syncope. PMID- 12082061 TI - Acute hyperperfusion syndrome complicating EC-IC bypass. PMID- 12082062 TI - A Japanese case of steroid responsive myopathy with deficient chondroitin sulphate. PMID- 12082064 TI - Palatal tremor and cognitive decline in neuroferritinopathy. PMID- 12082063 TI - Fulminant progression of hyperammonaemic encephalopathy after treatment with valproate in a patient with ureterosigmoidostomy. PMID- 12082065 TI - Sulcal abnormalities on brain magnetic resonance imaging in the Guillain-Barre syndrome. PMID- 12082067 TI - Muscle tissue oxygenation as a functional tool in the follow up of dermatomyositis. PMID- 12082066 TI - Cocaine induced hypokalaemic periodic paralysis. PMID- 12082068 TI - Propofol in myoclonus status epilepticus in comatose patients following cardiac resuscitation. PMID- 12082069 TI - Chronic asymmetric progressive external ophthalmoplegia with right facial weakness: a unique presentation of mitochondrial myopathy. PMID- 12082070 TI - Thunderclap headache, reversible cerebral arterial vasoconstriction, and unruptured aneurysms. PMID- 12082071 TI - Radiology of stroke. PMID- 12082072 TI - Cochlear implantation in a profoundly deaf patient with MELAS syndrome. PMID- 12082073 TI - Bilateral lesions restricted to the posteroventral pallidum are unlikely to provoke corticobulbar syndrome and psychic akinesia. PMID- 12082075 TI - Cell cycle-dependent localization of macroH2A in chromatin of the inactive X chromosome. AB - One of several features acquired by chromatin of the inactive X chromosome (Xi) is enrichment for the core histone H2A variant macroH2A within a distinct nuclear structure referred to as a macrochromatin body (MCB). In addition to localizing to the MCB, macroH2A accumulates at a perinuclear structure centered at the centrosome. To better understand the association of macroH2A1 with the centrosome and the formation of an MCB, we investigated the distribution of macroH2A1 throughout the somatic cell cycle. Unlike Xi-specific RNA, which associates with the Xi throughout interphase, the appearance of an MCB is predominantly a feature of S phase. Although the MCB dissipates during late S phase and G2 before reforming in late G1, macroH2A1 remains associated during mitosis with specific regions of the Xi, including at the X inactivation center. This association yields a distinct macroH2A banding pattern that overlaps with the site of histone H3 lysine-4 methylation centered at the DXZ4 locus in Xq24. The centrosomal pool of macroH2A1 accumulates in the presence of an inhibitor of the 20S proteasome. Therefore, targeting of macroH2A1 to the centrosome is likely part of a degradation pathway, a mechanism common to a variety of other chromatin proteins. PMID- 12082076 TI - The roles of Fzy/Cdc20 and Fzr/Cdh1 in regulating the destruction of cyclin B in space and time. AB - In Drosophila cells cyclin B is normally degraded in two phases: (a) destruction of the spindle-associated cyclin B initiates at centrosomes and spreads to the spindle equator; and (b) any remaining cytoplasmic cyclin B is degraded slightly later in mitosis. We show that the APC/C regulators Fizzy (Fzy)/Cdc20 and Fzy related (Fzr)/Cdh1 bind to microtubules in vitro and associate with spindles in vivo. Fzy/Cdc20 is concentrated at kinetochores and centrosomes early in mitosis, whereas Fzr/Cdh1 is concentrated at centrosomes throughout the cell cycle. In syncytial embryos, only Fzy/Cdc20 is present, and only the spindle-associated cyclin B is degraded at the end of mitosis. A destruction box-mutated form of cyclin B (cyclin B triple-point mutant [CBTPM]-GFP) that cannot be targeted for destruction by Fzy/Cdc20, is no longer degraded on spindles in syncytial embryos. However, CBTPM-GFP can be targeted for destruction by Fzr/Cdh1. In cellularized embryos, which normally express Fzr/Cdh1, CBTPM-GFP is degraded throughout the cell but with slowed kinetics. These findings suggest that Fzy/Cdc20 is responsible for catalyzing the first phase of cyclin B destruction that occurs on the mitotic spindle, whereas Fzr/Cdh1 is responsible for catalyzing the second phase of cyclin B destruction that occurs throughout the cell. These observations have important implications for the mechanisms of the spindle checkpoint. PMID- 12082077 TI - Transmission of cell stress from endoplasmic reticulum to mitochondria: enhanced expression of Lon protease. AB - The rat homologue of a mitochondrial ATP-dependent protease Lon was cloned from cultured astrocytes exposed to hypoxia. Expression of Lon was enhanced in vitro by hypoxia or ER stress, and in vivo by brain ischemia. These observations suggested that changes in nuclear gene expression (Lon) triggered by ER stress had the potential to impact important mitochondrial processes such as assembly and/or degradation of cytochrome c oxidase (COX). In fact, steady-state levels of nuclear-encoded COX IV and V were reduced, and mitochondrial-encoded subunit II was rapidly degraded under ER stress. Treatment of cells with cycloheximide caused a similar imbalance in the accumulation of COX subunits, and enhanced mRNA for Lon and Yme1, the latter another mitochondrial ATP-dependent protease. Furthermore, induction of Lon or GRP75/mtHSP70 by ER stress was inhibited in PERK (-/-) cells. Transfection studies revealed that overexpression of wild-type or proteolytically inactive Lon promoted assembly of COX II into a COX I-containing complex, and partially prevented mitochondrial dysfunction caused by brefeldin A or hypoxia. These observations demonstrated that suppression of protein synthesis due to ER stress has a complex effect on the synthesis of mitochondrial associated proteins, both COX subunits and ATP-dependent proteases and/or chaperones contributing to assembly of the COX complex. PMID- 12082078 TI - PRC1 is a microtubule binding and bundling protein essential to maintain the mitotic spindle midzone. AB - Midzone microtubules of mammalian cells play an essential role in the induction of cell cleavage, serving as a platform for a number of proteins that play a part in cytokinesis. We demonstrate that PRC1, a mitotic spindle-associated Cdk substrate that is essential to cell cleavage, is a microtubule binding and bundling protein both in vivo and in vitro. Overexpression of PRC1 extensively bundles interphase microtubules, but does not affect early mitotic spindle organization. PRC1 contains two Cdk phosphorylation motifs, and phosphorylation is possibly important to mitotic suppression of bundling, as a Cdk phosphorylation-null mutant causes extensive bundling of the prometaphase spindle. Complete suppression of PRC1 by siRNA causes failure of microtubule interdigitation between half spindles and the absence of a spindle midzone. Truncation mutants demonstrate that the NH2-terminal region of PRC1, rich in alpha-helical sequence, is important for localization to the cleavage furrow and to the center of the midbody, whereas the central region, with the highest sequence homology between species, is required for microtubule binding and bundling activity. We conclude that PRC1 is a microtubule-associated protein required to maintain the spindle midzone, and that distinct functions are associated with modular elements of the primary sequence. PMID- 12082079 TI - MAP2 and tau bind longitudinally along the outer ridges of microtubule protofilaments. AB - MAP2 and tau exhibit microtubule-stabilizing activities that are implicated in the development and maintenance of neuronal axons and dendrites. The proteins share a homologous COOH-terminal domain, composed of three or four microtubule binding repeats separated by inter-repeats (IRs). To investigate how MAP2 and tau stabilize microtubules, we calculated 3D maps of microtubules fully decorated with MAP2c or tau using cryo-EM and helical image analysis. Comparing these maps with an undecorated microtubule map revealed additional densities along protofilament ridges on the microtubule exterior, indicating that MAP2c and tau form an ordered structure when they bind microtubules. Localization of undecagold attached to the second IR of MAP2c showed that IRs also lie along the ridges, not between protofilaments. The densities attributable to the microtubule-associated proteins lie in close proximity to helices 11 and 12 and the COOH terminus of tubulin. Our data further suggest that the evolutionarily maintained differences observed in the repeat domain may be important for the specific targeting of different repeats to either alpha or beta tubulin. These results provide strong evidence suggesting that MAP2c and tau stabilize microtubules by binding along individual protofilaments, possibly by bridging the tubulin interfaces. PMID- 12082080 TI - L1 endocytosis is controlled by a phosphorylation-dephosphorylation cycle stimulated by outside-in signaling by L1. AB - Dynamic regulation of the cell surface expression of adhesion molecules is an important mechanism for controlling neuronal growth cone motility and guidance. Clathrin-mediated vesicular internalization of L1 via the tyrosine-based endocytosis motif YRSL regulates adhesion and signaling by this Ig superfamily molecule. Here, we present evidence that tyrosine-1176 (Y1176) of the YRSL motif is phosphorylated in vivo. The nonreceptor tyrosine kinase (p60src) is implicated in L1-mediated neurite outgrowth, and we find that p60src phosphorylates Y1176 in vitro. Phosphorylation of Y1176 prevents L1 binding to AP-2, an adaptor required for clathrin-mediated internalization of L1. mAb 74-5H7 recognizes the sequence immediately NH2-terminal to the tyrosine-based motif and binds L1 only when Y1176 is dephosphorylated. 74-5H7 identifies a subset of L1 present at points of cell cell contact and in vesicle-like structures that colocalize with an endocytosis marker. L1-L1 binding or L1 cross-linking induces a rapid increase in 74-5H7 immunoreactivity. Our data suggest a model in which homophilic binding or L1 cross-linking triggers transient dephosphorylation of the YRSL motif that makes L1 available for endocytosis. Thus, the regulation of L1 endocytosis through dephosphorylation of Y1176 is a critical regulatory point of L1-mediated adhesion and signaling. PMID- 12082081 TI - Dynamic interaction of VCAM-1 and ICAM-1 with moesin and ezrin in a novel endothelial docking structure for adherent leukocytes. AB - Ezrin, radixin, and moesin (ERM) regulate cortical morphogenesis and cell adhesion by connecting membrane adhesion receptors to the actin-based cytoskeleton. We have studied the interaction of moesin and ezrin with the vascular cell adhesion molecule (VCAM)-1 during leukocyte adhesion and transendothelial migration (TEM). VCAM-1 interacted directly with moesin and ezrin in vitro, and all of these molecules colocalized at the apical surface of endothelium. Dynamic assessment of this interaction in living cells showed that both VCAM-1 and moesin were involved in lymphoblast adhesion and spreading on the endothelium, whereas only moesin participated in TEM, following the same distribution pattern as ICAM-1. During leukocyte adhesion in static or under flow conditions, VCAM-1, ICAM-1, and activated moesin and ezrin clustered in an endothelial actin-rich docking structure that anchored and partially embraced the leukocyte containing other cytoskeletal components such as alpha-actinin, vinculin, and VASP. Phosphoinositides and the Rho/p160 ROCK pathway, which participate in the activation of ERM proteins, were involved in the generation and maintenance of the anchoring structure. These results provide the first characterization of an endothelial docking structure that plays a key role in the firm adhesion of leukocytes to the endothelium during inflammation. PMID- 12082082 TI - Retention of a cell adhesion complex at the paranodal junction requires the cytoplasmic region of Caspr. AB - An axonal complex of cell adhesion molecules consisting of Caspr and contactin has been found to be essential for the generation of the paranodal axo-glial junctions flanking the nodes of Ranvier. Here we report that although the extracellular region of Caspr was sufficient for directing it to the paranodes in transgenic mice, retention of the Caspr-contactin complex at the junction depended on the presence of an intact cytoplasmic domain of Caspr. Using immunoelectron microscopy, we found that a Caspr mutant lacking its intracellular domain was often found within the axon instead of the junctional axolemma. We further show that a short sequence in the cytoplasmic domain of Caspr mediated its binding to the cytoskeleton-associated protein 4.1B. Clustering of contactin on the cell surface induced coclustering of Caspr and immobilized protein 4.1B at the plasma membrane. Furthermore, deletion of the protein 4.1B binding site accelerated the internalization of a Caspr-contactin chimera from the cell surface. These results suggest that Caspr serves as a "transmembrane scaffold" that stabilizes the Caspr/contactin adhesion complex at the paranodal junction by connecting it to cytoskeletal components within the axon. PMID- 12082083 TI - Proximal location of mouse prostate epithelial stem cells: a model of prostatic homeostasis. AB - Stem cells are believed to regulate normal prostatic homeostasis and to play a role in the etiology of prostate cancer and benign prostatic hyperplasia. We show here that the proximal region of mouse prostatic ducts is enriched in a subpopulation of epithelial cells that exhibit three important attributes of epithelial stem cells: they are slow cycling, possess a high in vitro proliferative potential, and can reconstitute highly branched glandular ductal structures in collagen gels. We propose a model of prostatic homeostasis in which mouse prostatic epithelial stem cells are concentrated in the proximal region of prostatic ducts while the transit-amplifying cells occupy the distal region of the ducts. This model can account for many biological differences between cells of the proximal and distal regions, and has implications for prostatic disease formation. PMID- 12082084 TI - Dominant regulation of interendothelial cell gap formation by calcium-inhibited type 6 adenylyl cyclase. AB - Acute transitions in cytosolic calcium ([Ca2+]i) through store-operated calcium entry channels catalyze interendothelial cell gap formation that increases permeability. However, the rise in [Ca2+]i only disrupts barrier function in the absence of a rise in cAMP. Discovery that type 6 adenylyl cyclase (AC6; EC 4.6.6.1) is inhibited by calcium entry through store-operated calcium entry pathways provided a plausible explanation for how inflammatory [Ca2+]i mediators may decrease cAMP necessary for endothelial cell gap formation. [Ca2+]i mediators only modestly decrease global cAMP concentrations and thus, to date, the physiological role of AC6 is unresolved. Present studies used an adenoviral construct that expresses the calcium-stimulated AC8 to convert normal calcium inhibition into stimulation of cAMP, within physiologically relevant concentration ranges. Thrombin stimulated a dose-dependent [Ca2+]i rise in both pulmonary artery (PAECs) and microvascular (PMVEC) endothelial cells, and promoted intercellular gap formation in both cell types. In PAECs, gap formation was progressive over 2 h, whereas in PMVECs, gap formation was rapid (within 10 min) and gaps resealed within 2 h. Expression of AC8 resulted in a modest calcium stimulation of cAMP, which virtually abolished thrombin-induced gap formation in PMVECs. Findings provide the first direct evidence that calcium inhibition of AC6 is essential for endothelial gap formation. PMID- 12082086 TI - cGMP-dependent protein kinase I beta physically and functionally interacts with the transcriptional regulator TFII-I. AB - Transcriptional regulation of the fos promoter by nitric oxide and cGMP can occur by nuclear translocation of cGMP-dependent protein kinase I (G-kinase I) (Gudi, T., Lohmann, S. M., and Pilz, R. B. (1997) Mol. Cell. Biol. 17, 5244-5254). To identify nuclear targets of G-kinase I, we performed a yeast two-hybrid screen with G-kinase I beta as bait. We found that G-kinase I beta interacted specifically with TFII-I, an unusual transcriptional regulator that associates with multiple proteins to modulate both basal and signal-induced transcription. By using purified recombinant proteins, the interaction was mapped to the N terminal 93 amino acids of G-kinase I beta and one of six 95-amino acid repeats found in TFII-I. In baby hamster kidney cells, cGMP analogs enhanced co immunoprecipitation of G-kinase I beta and TFII-I by inducing co-localization of both proteins in the nucleus, but in other cell types containing cytoplasmic TFII I the G-kinase-TFII-I interaction was largely cGMP-independent. G-kinase phosphorylated TFII-I in vitro and in vivo on Ser(371) and Ser(743) outside of the interaction domain. G-kinase strongly enhanced TFII-I transactivation of a serum-response element-containing promoter in COS7 cells, and this effect was lost when Ser(371) and Ser(743) of TFII-I were mutated. TFII-I by itself had little effect on a full-length fos promoter in baby hamster kidney cells, but it synergistically enhanced transcriptional activation by G-kinase I beta. Binding of G-kinase to TFII-I may position the kinase to phosphorylate and regulate TFII I and/or factors that interact with TFII-I at the serum-response element. PMID- 12082085 TI - Matrix assembly, regulation, and survival functions of laminin and its receptors in embryonic stem cell differentiation. AB - Laminin-1 is essential for early embryonic basement membrane assembly and differentiation. Several steps can be distinguished, i.e., the expression of laminin and companion matrix components, their accumulation on the cell surface and assembly into basement membrane between endoderm and inner cell mass, and the ensuing differentiation of epiblast. In this study, we used differentiating embryoid bodies derived from mouse embryonic stem cells null for gamma1-laminin, beta1-integrin and alpha/beta-dystroglycan to dissect the contributions of laminin domains and interacting receptors to this process. We found that (a) laminin enables beta1-integrin-null embryoid bodies to assemble basement membrane and achieve epiblast with beta1-integrin enabling expression of the laminin alpha1 subunit; (b) basement membrane assembly and differentiation require laminin polymerization in conjunction with cell anchorage, the latter critically dependent upon a heparin-binding locus within LG module-4; (c) dystroglycan is not uniquely required for basement membrane assembly or initial differentiation; (d) dystroglycan and integrin cooperate to sustain survival of the epiblast and regulate laminin expression; and (e) laminin, acting via beta1-integrin through LG1-3 and requiring polymerization, can regulate dystroglycan expression. PMID- 12082087 TI - Complex role of the beta 2-beta 3 loop in the interaction of U1A with U1 hairpin II RNA. AB - RNA recognition motifs (RRMs) are characterized by highly conserved regions located centrally on a beta-sheet, which forms the RNA binding surface. Variable flanking regions, such as the loop connecting beta-strands 2 and 3, are thought to be important in determining the RNA-binding specificities of individual RRMs. The N-terminal RRM of the spliceosomal U1A protein mediates binding to an RNA hairpin (U1hpII) in the U1 small nuclear RNA. In this complex, the beta(2) beta(3) loop protrudes through the 10-nucleotide RNA loop. Shortening of the RNA loop strongly perturbs binding, suggesting that an optimal "fit" of the beta(2) beta(3) loop into the RNA loop is an important factor in complexation. To understand this interaction further, we mutated or deleted loop residues Lys(50) and Met(51), which protrude centrally into the RNA loop but do not make any direct contacts to the bases. Using BIACORE, we analyzed the ability of these U1A mutants to bind to wild type RNAs, or RNAs with shortened loops. Alanine replacement mutations only modestly affected binding to wild type U1hpII. Interestingly, simultaneous replacement of Lys(50) and Met(51) with alanine appeared to alleviate the loss of binding caused by shortening of the RNA loop. Deletion of Lys(50) or Met(51) caused a dramatic loss in stability of the U1A.U1hpII complex. However, deletion of both residues simultaneously was much less deleterious. Simulated annealing molecular dynamics analyses suggest this is due to the ability of this mutant to rearrange flanking amino acids to substitute for the two deleted residues. The double deletion mutant also exhibited substantially reduced negative effects of RNA loop shortening, suggesting the rearranged loop is better able to accommodate a short RNA loop. Our results indicate that one of the roles of the beta(2)-beta(3) loop is to provide a steric fit into the RNA loop, thereby stabilizing the RNA.protein complex. PMID- 12082088 TI - Up-regulation of sodium-dependent glucose transporter by interaction with heat shock protein 70. AB - Heat shock stress induces some heat shock proteins, including Hsp70, and activates sodium-dependent glucose transport in porcine renal LLC-PK(1) cells, but its mechanisms have not been described in detail. We investigated whether sodium-dependent glucose transporter (SGLT1) interacts with Hsp70 to increase SGLT1 activity. Heat shock stress increased SGLT1 activity without changing SGLT1 expression. The increase of SGLT1 activity was completely inhibited by an anti transforming growth factor-beta1 (TGF-beta1) antibody. Instead of heat shock stress, TGF-beta1 increased SGLT1 activity dose- and time-dependently without changing SGLT1 expression. We found that the amount of Hsp70 immunoprecipitated from TGF-beta1-treated cells with an anti-SGLT1 antibody was higher than that of the control cells. Transfection of an anti-Hsp70 antibody into the cells inhibited the increase of SGLT1 activity. With confocal laser microscopy, both SGLT1 and Hsp70 was localized near the apical membrane in the TGF-beta1-treated cells, and an anti-Hsp70 antibody disturbed this localization. Furthermore, we clarified that an anti-Hsp70 antibody inhibited interaction of SGLT1 with Hsp70 in vitro. These results suggest that Hsp70 forms a complex with SGLT1 and increases the expression level of SGLT1 in the apical membrane, resulting in up regulation of glucose uptake. PMID- 12082089 TI - The cleavage/polyadenylation activity triggered by a U-rich motif sequence is differently required depending on the poly(A) site location at either the first or last 3'-terminal exon of the 2'-5' oligo(A) synthetase gene. AB - Production of the two mRNAs encoding distinct forms of 2'-5'-oligoadenylate synthetase depends on processing that involves the recognition of alternative poly(A) sites and an internal 5'-splice site located within the first 3'-terminal exon. The resulting 1.6- and 1.8-kb mRNAs are expressed in fibroblast cell lines, whereas lymphoblastoid B cells, such as Daudi, produce only the 1.8-kb mRNA. In the present study, we have shown that the 3'-end processing at the last 3' terminal exon occurs independently of the core poly(A) site sequence or the presence of regulatory elements. In contrast, in Daudi cells, the recognition of the poly(A) site at the first 3'-terminal exon is impaired because of an unfavorable sequence context. The 3'-end processing at this particular location requires a strong stabilization of the cleavage/polyadenylation factors, which can be achieved by the insertion of a 25-nucleotide long U-rich motif identified upstream of the last poly(A) site. Consequently, we speculate that in cells expressing the 1.6-kb mRNA, such as fibroblasts, direct or indirect participation of a specific mechanism or cell type-specific factors are required for an efficient polyadenylation at the first 3'-terminal exon. PMID- 12082091 TI - BRCA1-induced apoptosis involves inactivation of ERK1/2 activities. AB - Mutation in the BRCA1 gene is associated with an increased risk of breast and ovarian cancer. Recent studies have shown that the BRCA1 gene product may be important in mediating responses to DNA damage and genomic instability. Previous studies have indicated that overexpression of BRCA1 can induce apoptosis or cell cycle arrest at the G(2)/M border in various cell types. Although the activation of JNK kinase has been implicated in BRCA1-induced apoptosis, the role of other members of the mitogen-activated protein kinase family in mediating the cellular response to BRCA1 has not yet been examined. In this study, we monitored the activities of three members of the MAPK family (ERK1/2, JNK, p38) in MCF-7 breast cancer cells and U2OS osteosarcoma cells after their exposure to a recombinant adenovirus expressing wild type BRCA1 (Ad.BRCA1). Overexpression of BRCA1 in MCF 7 cells resulted in arrest at the G(2)/M border; however, BRCA1 expression in U2OS cells induced apoptosis. Although BRCA1 induced JNK activation in both cell lines, there were marked differences in ERK1/2 activation in response to BRCA1 expression in these two cell lines. BRCA1-induced apoptosis in U2OS cells was associated with no activation of ERK1/2. In contrast, BRCA1 expression in MCF-7 cells resulted in the activation of both ERK1/2 and JNK. To directly assess the role of ERK1/2 in determining the cellular response to BRCA1, we used dominant negative mutants of MEK1 as well as MEK1/2 inhibitor PD98059. Our results indicate that inhibition of ERK1/2 activation resulted in increased apoptosis after BRCA1 expression in MCF-7 cells. Furthermore, BRCA1-induced apoptosis involved activation of JNK, induction of Fas-L/Fas interaction, and activation of caspases 8 and 9. The studies presented in this report indicate that the response to BRCA1 expression is determined by the regulation of both the JNK and ERK1/2 signaling pathways in cells. PMID- 12082090 TI - MAP kinase stimulation by cAMP does not require RAP1 but SRC family kinases. AB - The small G protein RAP1 and the kinase B-RAF have been proposed to link elevations of cAMP to activation of ERK/mitogen-activated protein (MAP) kinase. In order to delineate signaling pathways that link receptor-generated cAMP to the activation of MAP kinase, the human A(2A)-adenosine receptor, a prototypical G(s) coupled receptor, was heterologously expressed in Chinese hamster ovary cells (referred as CHO-A(2A) cells). In CHO-A(2A) cells, the stimulation of the A(2A) receptor resulted in an activation of RAP1 and formation of RAP1-B-RAF complexes. However, overexpression of a RAP1 GTPase-activating protein (RAP1GAP), which efficiently clamped cellular RAP1 in the inactive GDP-bound form, did not affect A(2A)-agonist-mediated MAP kinase stimulation. In contrast, the inhibitor of protein kinase A H89 efficiently suppressed A(2A)-agonist-mediated MAP kinase stimulation. Neither dynamin-dependent receptor internalization nor receptor promoted shedding of matrix-bound growth factors accounted for A(2A)-receptor dependent MAP kinase activation. PP1, an inhibitor of SRC family kinases, blunted both the A(2A)-receptor- and the forskolin-induced MAP kinase stimulation (IC(50) = 50 nm); this was also seen in PC12 cells, which express the A(2A)-receptor endogenously, and in NIH3T3 fibroblasts, in which cAMP causes MAP kinase stimulation. In the corresponding murine fibroblast cell line SYF, which lacks the ubiquitously expressed SRC family kinases SRC, YES, and FYN, forskolin barely stimulated MAP kinase; this reduction was reversed in cells in which c-SRC had been reintroduced. These findings show that activation of MAP kinase by cAMP requires a SRC family kinase that lies downstream of protein kinase A. A role for RAP1, as documented for the beta(2)-adrenergic receptor, is apparently contingent on receptor endocytosis. PMID- 12082092 TI - Isolation and analysis of mutant alleles of the Bacillus subtilis HrcA repressor with reduced dependency on GroE function. AB - The hrcA gene of Bacillus subtilis codes for a transcriptional repressor protein that negatively regulates expression of the heptacistronic dnaK and the bicistronic groE operon by binding to an operator-element called CIRCE. Recently, we have published data suggesting that the activity of HrcA is modulated by the GroE chaperonin system. Biochemical analyses of the HrcA protein have been hampered so far by its strong tendency to aggregate. Here, a genetic method was used to isolate mutant forms of HrcA with increased activity under conditions of decreased GroE function. One of these mutant forms (HrcA114) containing five amino acid replacements exhibited enhanced solubility when overexpressed. HrcA114 purified under native conditions produced two retarded CIRCE-containing DNA fragments in band shift experiments. The amount of the larger fragment increased after addition of GroEL, GroES, and ATP but decreased when ATP was replaced by the nonhydrolyzable ATP analog ATPgammaS. DNase I footprinting experiments exhibited full protection of the CIRCE element and neighboring nucleotides in an asymmetric way. An in vitro binding assay using affinity chromatography showed direct and specific interaction between HrcA114 and GroEL. All these experimental data are in full agreement with our previously published model that HrcA needs the GroE chaperonin system for activation. PMID- 12082093 TI - Dynamitin controls Beta 2 integrin avidity by modulating cytoskeletal constraint on integrin molecules. AB - Dynamitin, a subunit of the microtubule-dependent motor complex, was implicated in cell adhesion by binding to MacMARCKS (Macrophage-enriched myristoylated alanine-rice C kinase substrate). However, how dynamitin is involved in cell adhesion is unclear despite the fact that both MacMARCKS and microtubules regulate beta(2) integrin activation. We report that dynamitin regulates beta(2) integrin avidity toward iC3b by modulating the lateral mobility of beta(2) integrin molecules. Using the single particle tracking method, we found that integrin molecular mobility in cells expressing the fusion protein CFP (cyan fluorescent protein)-dynamitin or CFP-MB (the MacMARCKS binding domain peptide of dynamitin) increased 6-fold over the control cells, suggesting that disturbing dynamitin function dramatically altered the cytoskeletal constraint on beta(2) integrin molecules. Further mechanistic studies revealed that overexpression of dynamitin stimulated the phosphorylation of endogenous MacMARCKS protein, which lead to the enhanced tyrosine phosphorylation of paxillin. This effect of dynamitin correlates with the observation that higher concentration of PKC inhibitor is required to block beta(2) integrin mobility in dynamitin-expressing cells. Although dynamitin acts at the point of MacMARCKS phosphorylation, it is upstream of RhoA, because its effect was blocked by RhoA inhibitor. Thus, we conclude that dynamitin is a part of the cytoskeletal constraint that locks beta(2) integrin in the inactive form. PMID- 12082094 TI - Functions of transforming growth factor-beta family type I receptors and Smad proteins in the hypertrophic maturation and osteoblastic differentiation of chondrocytes. AB - We investigated the effects of bone morphogenetic protein (BMP)-2, a member of the transforming growth factor-beta superfamily, on the regulation of the chondrocyte phenotype, and we identified signaling molecules involved in this regulation. BMP-2 triggers three concomitant responses in mouse primary chondrocytes and chondrocytic MC615 cells. First, BMP-2 stimulates expression or synthesis of type II collagen. Second, BMP-2 induces expression of molecular markers characteristic of pre- and hypertrophic chondrocytes, such as Indian hedgehog, parathyroid hormone/parathyroid hormone-related peptide receptor, type X collagen, and alkaline phosphatase. Third, BMP-2 induces osteocalcin expression, a specific trait of osteoblasts. Constitutively active forms of transforming growth factor-beta family type I receptors and Smad proteins were overexpressed to address their role in this process. Activin receptor-like kinase (ALK)-1, ALK-2, ALK-3, and ALK-6 were able to reproduce the hypertrophic maturation of chondrocytes induced by BMP-2. In addition, ALK-2 mimicked further the osteoblastic differentiation of chondrocytes induced by BMP-2. In the presence of BMP-2, Smad1, Smad5, and Smad8 potentiated the hypertrophic maturation of chondrocytes, but failed to induce osteocalcin expression. Smad6 and Smad7 impaired chondrocytic expression and osteoblastic differentiation induced by BMP-2. Thus, our results indicate that Smad-mediated pathways are essential for the regulation of the different steps of chondrocyte and osteoblast differentiation and suggest that additional Smad-independent pathways might be activated by ALK-2. PMID- 12082095 TI - Interaction of p58(PITSLRE), a G2/M-specific protein kinase, with cyclin D3. AB - The p58(PITSLRE) is a p34(cdc2)-related protein kinase that plays an important role in normal cell cycle progression. Elevated expression of p58(PITSLRE) in eukaryotic cells prevents them from undergoing normal cytokinesis and appears to delay them in late telophase. To investigate the molecular mechanism of p58(PITSLRE) action, we used the yeast two-hybrid system, screened a human fetal liver cDNA library, and identified cyclin D3 as an interacting partner of p58(PITSLRE). In vitro binding assay, in vivo coimmunoprecipitation, and immunofluorescence cell staining further confirmed the association of p58(PITSLRE) with cyclin D3. This binding was observed only in the G(2)/M phase but not in the G(1)/S phase of the cell cycle; meanwhile, no interaction between p110(PITSLRE) and cyclin D3 was observed in all the cell cycle. The overexpression of cyclin D3 in 7721 cells leads to an exclusively accumulation of p58(PITSLRE) in the nuclear region, affecting its cellular distribution. Histone H1 kinase activity of p58(PITSLRE) was greatly enhanced upon interaction with cyclin D3. Furthermore, kinase activity of p58(PITSLRE) was found to increase greatly in the presence of cyclin D3 using a specific substrate, beta-1,4 galactosyltransferase 1. These data provide a new clue to our understanding of the cellular function of p58(PITSLRE) and cyclin D3. PMID- 12082096 TI - Membrane topology of the hepatitis C virus NS2 protein. AB - The hepatitis C virus (HCV) NS2 protein is a hydrophobic protein. Previous studies indicate that this protein is an integral membrane protein, which is targeted to the membrane of the endoplasmic reticulum (ER) by the signal sequence located in its preceding p7 protein. In this report, we demonstrate that the membrane association of NS2 is p7-independent and occurs co-translationally. Further deletion-mapping studies suggest the presence of two internal signal sequences in NS2. These two internal signal sequences, which are located within amino acids 839-883 and amino acids 928-960, could target the alpha-globin reporter, a cytosolic protein, to the membrane compartments in HuH7 hepatoma cells. The presence of multiple signal sequences for its membrane association suggests that NS2 has multiple transmembrane domains. The glycosylation studies indicate that both amino and carboxyl termini of NS2 are located in the endoplasmic reticulum lumen. Based on these results, a model for the NS2 membrane topology is presented. PMID- 12082097 TI - Osmoregulation of endothelial nitric-oxide synthase gene expression in inner medullary collecting duct cells. Role in activation of the type A natriuretic peptide receptor. AB - Previously, we showed that increased extracellular tonicity promotes increased type A natriuretic peptide receptor (NPR-A) expression through a p38 MAPKbeta pathway in inner medullary collecting duct cells. The endothelial and inducible nitric-oxide synthase (eNOS and iNOS respectively) genes are also expressed in this nephron segment and are thought to play a role in regulating urinary sodium concentration. We sought to determine whether changes in tonicity might regulate NOS gene expression, and if so, whether these latter changes might be linked mechanistically to the increase in NPR-A gene expression. Increased extracellular tonicity effected a time-dependent reduction in eNOS and iNOS protein levels, eNOS mRNA levels, and eNOS gene promoter activity over the first 8 h of the incubation. Although levels of the eNOS mRNA and promoter activity had returned to normal after 24 h, eNOS protein levels remained low at 24-36 h, and recovery was not complete even at 48 h. The decrease in eNOS expression was signaled in large part through a p38 MAPK-dependent mechanism. Reduction in eNOS expression together with the concomitant decline in intracellular cyclic GMP levels appears to account for a significant portion of the p38 MAPK-dependent osmotic stimulation of NPR-A gene expression noted previously. Collectively, these findings support the existence of a complex regulatory circuitry in the cells of the inner medullary collecting duct linking two independent cyclic GMP-generating signal transduction systems involved in regulation of urinary sodium concentration. PMID- 12082098 TI - The apoptotic protein tBid promotes leakage by altering membrane curvature. AB - The apoptotic protein tBid is effective in promoting both leakage and lipid mixing in liposomes composed of cardiolipin and phosphatidylcholine at a molar ratio of 1:2 in the presence of calcium. When half of the phosphatidylcholine component of these liposomes is replaced with phosphatidylethanolamine, a lipid that promotes negative membrane curvature, the rates of both leakage and lipid mixing caused by tBid are substantially increased. Replacement of cardiolipin with phosphatidylglycerol, a lipid that is structurally similar to cardiolipin but does not promote negative membrane curvature in the presence of calcium, prevents the tBid from promoting leakage. The promotion of leakage by tBid is also inhibited by several substances that promote positive membrane curvature, including lysophosphatidylcholine, tritrpticin, a potent antimicrobial peptide, and cyclosporin A, a known inhibitor of cytochrome c release from mitochondria. We directly measured the effect of tBid on membrane curvature by (31)P NMR. We found that tBid promotes the formation of highly curved non-lamellar phases. All of these data are consistent with the hypothesis that tBid promotes negative curvature, and as a result it destabilizes bilayer membranes. Bcl-X(L) inhibits leakage and lipid mixing induced by tBid. Bcl-X(L) is anti-apoptotic. It reduces the promotion of non-bilayer phases by tBid, although by itself Bcl-X(L) is capable of promoting their formation. Bcl-X(L) has little effect on liposomal integrity. Our results suggest that the anti-apoptotic activity of Bcl-X(L) is not a consequence of its interaction with membranes, but rather with other proteins, such as tBid. PMID- 12082099 TI - Solution structure of peptide toxins that block mechanosensitive ion channels. AB - Mechanosensitive channels (MSCs) play key roles in sensory processing and have been implicated as primary transducers for a variety of cellular responses ranging from osmosensing to gene expression. This paper presents the first structures of any kind known to interact specifically with MSCs. GsMTx-4 and GsMtx-2 are inhibitor cysteine knot peptides isolated from venom of the tarantula, Grammostola spatulata (Suchyna, T. M., Johnson, J. H., Hamer, K., Leykam, J. F., Gage, D. A., Clemo, H. F., Baumgarten, C. M., and Sachs, F. (2000) J. Gen. Physiol. 115, 583-598). Inhibition of cationic MSCs by the higher affinity GsMtx-4 (K(D) approximately 500 nm) reduced cell size in swollen and hypertrophic heart cells, swelling-activated currents in astrocytes, and stretch induced arrhythmias in the heart. Despite the relatively low affinity, no cross reactivity has been found with other channels. Using two-dimensional NMR spectroscopy, we determined the solution structure of GsMTx-4 and a lower affinity (GsMTx-2; K(D) approximately 6 microm) peptide from the same venom. The dominant feature of the two structures is a hydrophobic patch, utilizing most of the aromatic residues and surrounded with charged residues. The spatial arrangement of charged residues that are unique to GsMTx-4 and GsMTx-2 may underlie the selectivity of these peptides. PMID- 12082100 TI - Differential roles of insulin receptor substrates in the anti-apoptotic function of insulin-like growth factor-1 and insulin. AB - Insulin-like growth factor-1 (IGF-1) and insulin are known to prevent apoptosis. The signaling network of IGF-1 and insulin occurs via multiple pathways involving different insulin receptor substrates (IRSs). To define their roles in the anti apoptotic function of IGF-1 and insulin, we established brown pre-adipocyte cell lines from wild-type and IRS knockout (KO) animals. In response to 16 h of serum deprivation, IRS-1-deficient cells showed a significant decrease in response to IGF-1 protection from apoptosis, whereas no changes were observed in the IRS-2, IRS-3, or IRS-4 KO cells. Five hours after serum withdrawal, cells already began to undergo apoptosis. At this early time point, IGF-1 and insulin were able to protect both wild-type and IRS-1 KO cells from death by 85-90%. After a longer period of serum deprivation, the protective ability of insulin and IGF-1 was decreased, and this was especially reduced in the IRS-1 KO cells. Reconstitution of these cells with IRS-1, IRS-2, IRS-3, or IRS-1/IRS-2 chimeras restored the anti-apoptotic effects of IGF-1, whereas overexpression of IRS-4 had no effect at long time points and actually reduced the effect of IGF-1 at the short time point. The biochemical basis of the defect in anti-apoptosis was not dependent on phosphorylation of mitogen-activated protein kinase; whereas phosphoinositide 3 kinase activity was decreased by 30% in IRS-1 KO cells. Akt phosphorylation was slightly reduced in these cells. Phosphorylation of the transcription factors cAMP response element-binding protein and FKHR by IGF-1 and insulin was markedly reduced in IRS-1 KO cells. In addition, both IGF-1 and insulin prevented caspase 3 cleavage in the wild-type cells, and this effect was greatly reduced in the IRS 1-deficient cells. These findings suggest that the IRS proteins may play differential roles in the anti-apoptotic effects of IGF-1 and insulin in brown pre-adipocytes, with IRS-1 being predominant, possibly acting through caspase-3-, CREB-, and FKHR-dependent mechanisms. PMID- 12082101 TI - Ultraviolet-induced junD activation and apoptosis in myeloblastic leukemia ML-1 cells. AB - The exposure of mammalian cells to UV irradiation induces the expression of immediate early genes such as c-jun and c-fos and activates the transcription factors AP-1 and NF-kappaB. JunD is one of the three members of the Jun family and shares some functional characteristics with c-Jun. In the present study, we found that the exposure of myeloblastic leukemia ML-1 cells to UV light (UVC) caused a significant increase in junD mRNA expression within 5 min that persisted for a period of 3 h. The activation of protein kinase C (PKC) with 12-O tetradecaoylphorbol-13-acetate (TPA) also induced increases in junD expression similar to those of UV irradiation. In addition, UV irradiation- and TPA-induced increases in junD expression were completely abolished by GF-109203X, a PKC specific inhibitor. UV irradiation activated intracellular signaling pathways including extracellular regulated kinase-2 (Erk-2), c-Jun N-terminal kinases-1 (JNK-1), and p38. However, TPA-induced activation of PKC affected only Erk-2 activity, and GF-109203X (a PKC inhibitor) markedly suppressed UV-induced Erk-2 activation. To further investigate the effect of UV-induced Erk-2 activation on the expression of junD mRNA, cDNA encoding mitogen-activated protein kinase kinase (MEK1) was overexpressed in ML-1 cells. The overexpression of MEK1 enhanced substantially junD expression in response to UV or TPA. In contrast, the suppression of Erk activation with PD98059, a specific inhibitor of MEK1, inhibited UV- and TPA-induced junD mRNA expression, UV-induced increases in caspase-3 activities, and cell death. In addition, the overexpression of junD enhanced the UV irradiation-induced increases in caspase-3 activity and cell death. We conclude that UV irradiation-induced increases in junD expression in ML 1 cells are mediated through activation of the PKC-coupled Erk-2 signaling pathway and play an important role in ML-1 cell apoptosis. PMID- 12082102 TI - The prostacyclin analogue treprostinil blocks NFkappaB nuclear translocation in human alveolar macrophages. AB - Primary pulmonary hypertension (PPH) is characterized by increased pulmonary arterial pressure and vascular resistance. We and others have observed that inflammatory cytokines and infiltrates are present in the lung tissue, but the significance is uncertain. Treprostinil (TRE), a prostacyclin analogue with extended half-life and chemical stability, has shown promise in the treatment of PPH. We hypothesize that TRE might exert beneficial effects in PPH by antagonizing inflammatory cytokine production in the lung. Here we show that TRE dose-dependently inhibits inflammatory cytokine (tumor necrosis factor-alpha, interleukin-1beta, interleukin-6, and granulocyte macrophage colony-stimulating factor) secretion and gene expression by human alveolar macrophages. TRE blocks NFkappaB activation, but IkappaB-alpha phosphorylation and degradation are unaffected. Moreover, TRE does not affect the formation of the NFkappaB.DNA complex but blocks nuclear translocation of p65. These results are the first to illustrate the anti-cytokine actions of TRE in down-regulating NFkappaB, not through its inhibitory component or by direct binding but by blocking nuclear translocation. These data indicate that inflammatory mechanisms may be important in the pathogenesis of PPH and cytokine antagonism by blocking NFkappaB may contribute to the efficacy of TRE therapy in PPH. PMID- 12082103 TI - The activation function-1 domain of Nur77/NR4A1 mediates trans-activation, cell specificity, and coactivator recruitment. AB - Nur77/NR4A1 is an "orphan member" of the nuclear hormone receptor superfamily. Nur77 and its close relatives Nurr1 and NOR-1 bind as monomers to a consensus binding site, the nerve growth factor induced protein I-B (NGFI-B)-binding response element (NBRE). The Nur77/NURR1/NOR1 nuclear receptors are classified as immediate early response genes which are induced through multiple signal transduction pathways. They have been implicated in cell proliferation, differentiation, and apoptosis. However, the mechanism of coactivation and ligand independent trans-activation remains unclear. Hence we examined the molecular basis of Nur77-mediated cofactor recruitment and activation. We observed that Nur77 trans-activates gene expression in a cell-specific manner, and operates in an activation function-1 (AF-1)-dependent manner. The AB region encodes an uncommonly potent N-terminal AF-1 domain delimited to between amino acids 50 and 160 and is essential for the ligand-independent activation of gene expression. Steroid receptor coactivator-2 (SRC-2) modulates the activity of the N-terminal AF-1 domain. Moreover, SRC-2 dramatically potentiates the retinoid induced RXR dependent activation of the Nur77 ligand binding domain (LBD). Interestingly, the N-terminal AB region (not the LBD) facilitates coactivator recruitment and directly interacts with SRC, p300, PCAF, and DRIP-205. Consistent with this, homology modeling indicated that the Nur77 LBD coactivator binding cleft was substantially different from that of retinoic acid receptor gamma, a closely related AF-2-dependent receptor. In particular, the hydrophobic cleft characteristic of nuclear receptors was replaced with a much more hydrophilic surface with a distinct topology. This observation accounts for the inability of this nuclear receptor LBD to directly mediate cofactor recruitment. Furthermore, the AF-1 domain physically associates with the Nur77 C-terminal LBD and synergizes with the retinoid X receptor LBD. Thus, the AF-1 domain plays a major role in Nur77-mediated transcriptional activation, cofactor recruitment, and intra- and intermolecular interactions. PMID- 12082104 TI - SNT1/FRS2 mediates germinal vesicle breakdown induced by an activated FGF receptor1 in Xenopus oocytes. AB - The docking protein SNT1/FRS2 (fibroblast growth factor receptor substrate 2) is implicated in the transmission of extracellular signals from the fibroblast growth factor receptor (FGFR), which plays vital roles during embryogenesis. Activating FGFR mutations cause several craniosynostoses and dwarfism syndromes in humans. Here we show that the Xenopus homolog of mammalian FRS-2 (XFRS2) is essential for the induction of oocyte maturation by an XFGFR1 harboring an activating mutation (XFGFR1act). Using a dominant-negative form of kinase suppressor of Ras, we show the Mek activity is required for germinal vesicle breakdown (GVBD) induced by co-expression of XFGFR1act and XFRS2, but this activity is not required for progesterone-induced GVBD. Furthermore, Mek/MAPK activity is critical for the induction and/or maintenance of H1 kinase activity at metaphase of meiosis II in progesterone-treated oocytes. An activated XFGFR1 containing a mutation in the phospholipase Cgamma binding site (XFGFR1actY672F) displayed a reduced ability to induce cell-cycle progression in oocytes, suggesting phospholipase Cgamma may not be necessary but that it augments XFGFR signaling in this system. Oocytes co-expressing XFGFR1act and XFRS2 showed substantial H1 kinase activity, but this activity was blocked when the oocytes were treated with the phosphatidylinositol 3-kinase inhibitor LY294002. Although phosphatidylinositol 3-kinase activity is essential for XFGFR1act/XFRS2-induced oocyte maturation, this activity is not required for maturation induced by progesterone. Finally, ectopic expression of Xspry2, a negative regulator of XFGFR signaling, greatly reduced MAPK activation and GVBD induced by the expression of either XFGFR1act plus XFRS2 or activated Ras (H-RasV12). In contrast, Xspry2 did not prevent GVBD induced by an activated form of Raf1, suggesting that Xspry2 exerts its inhibitory function upstream or parallel to Raf and downstream of Ras. PMID- 12082105 TI - The Three-dimensional structure of a superantigen-like protein, SET3, from a pathogenicity island of the Staphylococcus aureus genome. AB - The staphylococcal enterotoxin-like toxins (SETs) are a family of proteins encoded within the Staphylococcus aureus genome that were identified by their similarity to the well described bacterial superantigens. The first crystal structure of a member of the SET family, SET3, has been determined to 1.9 A (R = 0.205, R(free) = 0.240) and reveals a fold characteristic of the superantigen family but with significant differences. The SET proteins are secreted at varying levels by staphylococcal isolates, and seroconversion studies of normal individuals indicate that they are strongly antigenic to humans. Recombinant SETs do not exhibit any of the properties expected of superantigens such as major histocompatibility complex class II binding or broad T-cell activation, suggesting they have an entirely different function. The fact that the whole gene family is clustered within the pathogenicity island SaIn2 of the S. aureus genome suggests that they are involved in host/pathogen interactions. PMID- 12082106 TI - Raf-independent deregulation of p38 and JNK mitogen-activated protein kinases are critical for Ras transformation. AB - Activated Ras, but not Raf, causes transformation of RIE-1 epithelial cells, supporting the importance of Raf-independent pathways in mediating Ras transformation. The p38 and JNK mitogen-activated protein kinase cascades are activated by Ras via Raf-independent effector function. Therefore, we determined whether p38 and JNK activation are involved in Ras transformation of RIE-1 epithelial cells. Rather surprisingly, we found that pharmacologic inhibition of p38, together with Raf activation of ERK, was sufficient to mimic the morphologic and growth transformation caused by oncogenic Ras. p38 inhibition together with ERK activation also caused the same alterations in cyclin D1 and p21(CIP1) expression caused by Ras and induced an autocrine growth factor loop important for transformation. Finally, in contrast to p38, we found that JNK activation promoted Ras transformation, and that Ras deregulation of p38 and JNK was not mediated by activation of the Rac small GTPase. We conclude that a key action of Raf-independent effector pathways important for Ras transformation may involve inhibition of p38 and activation of JNK. PMID- 12082107 TI - The specificity of extracellular signal-regulated kinase 2 dephosphorylation by protein phosphatases. AB - The extracellular signal-regulated protein kinase 2 (ERK2) is the founding member of a family of mitogen-activated protein kinases (MAPKs) that are central components of signal transduction pathways for cell proliferation, stress responses, and differentiation. The MAPKs are unique among the Ser/Thr protein kinases in that they require both Thr and Tyr phosphorylation for full activation. The dual phosphorylation of Thr-183 and Tyr-185 in ERK2 is catalyzed by MAPK/ERK kinase 1 (MEK1). However, the identity and relative activity of protein phosphatases that inactivate ERK2 are less well established. In this study, we performed a kinetic analysis of ERK2 dephosphorylation by protein phosphatases using a continuous spectrophotometric enzyme-coupled assay that measures the inorganic phosphate produced in the reaction. Eleven different protein phosphatases, many previously suggested to be involved in ERK2 regulation, were compared, including tyrosine-specific phosphatases (PTP1B, CD45, and HePTP), dual specificity MAPK phosphatases (VHR, MKP3, and MKP5), and Ser/Thr protein phosphatases (PP1, PP2A, PP2B, PP2C alpha, and lambda PP). The results provide biochemical evidence that protein phosphatases display exquisite specificity in their substrate recognition and implicate HePTP, MKP3, and PP2A as ERK2 phosphatases. The fact that ERK2 inactivation could be carried out by multiple specific phosphatases shows that signals can be integrated into the pathway at the phosphatase level to determine the cellular response to external stimuli. Important insights into the roles of various protein phosphatases in ERK2 kinase signaling are obtained, and further analysis of the mechanism by which different protein phosphatases recognize and inactivate MAPKs will increase our understanding of how this kinase family is regulated. PMID- 12082108 TI - Deletion of the carboxyl terminus of Tie2 enhances kinase activity, signaling, and function. Evidence for an autoinhibitory mechanism. AB - Tie2 is an endothelial receptor tyrosine kinase that is required for both embryonic vascular development and tumor angiogenesis. There is considerable interest in understanding the mechanisms of Tie2 activation for therapeutic purposes. The recent solution of the Tie2 crystal structure suggests that Tie2 activity is autoinhibited by its carboxyl terminus. Here we investigated the role of the C tail in Tie2 activation, signaling, and function both in vitro and in vivo by deleting the C terminus of Tie2 (Delta CT). Compared to wild type Tie2, in vitro autophosphorylation and kinase activity were significantly enhanced by the Delta CT mutation. In NIH 3T3 cells expressing chimeric Tie2 receptors, both basal and ligand-induced tyrosine phosphorylation were markedly enhanced compared to wild type in several independent clones of Tie2-Delta CT. Moreover, the Delta CT mutation enhanced basal and ligand-dependent activation of Akt and extracellular signal-regulated kinase. Enhanced Akt activation correlated with significant inhibition of staurosporine-induced apoptosis. These findings demonstrate that the Tie2 C tail performs a novel negative regulatory role in Tie2 signaling and function, and they provide important insights into the mechanisms by which the Tie2 kinase is activated. PMID- 12082109 TI - Involvement of a bifunctional, paired-like DNA-binding domain and a transpositional enhancer in Sleeping Beauty transposition. AB - Sleeping Beauty (SB) is the most active Tc1/mariner-like transposon in vertebrate species. Each of the terminal inverted repeats (IRs) of SB contains two transposase-binding sites (DRs). This feature, termed the IR/DR structure, is conserved in a group of Tc1-like transposons. The DNA-binding region of SB transposase, similar to the paired domain of Pax proteins, consists of two helix turn-helix subdomains (PAI + RED = PAIRED). The N-terminal PAI subdomain was found to play a dominant role in contacting the DRs. Transposase was able to bind to mutant sites retaining the 3' part of the DRs; thus, primary DNA binding is not sufficient to determine the specificity of the transposition reaction. The PAI subdomain was also found to bind to a transpositional enhancer-like sequence within the left IR of SB, and to mediate protein-protein interactions between transposase subunits. A tetrameric form of the transposase was detected in solution, consistent with an interaction between the IR/DR structure and a transposase tetramer. We propose a model in which the transpositional enhancer and the PAI subdomain stabilize complexes formed by a transposase tetramer bound at the IR/DR. These interactions may result in enhanced stability of synaptic complexes, which might explain the efficient transposition of Sleeping Beauty in vertebrate cells. PMID- 12082110 TI - A novel antithrombotic role for high molecular weight kininogen as inhibitor of plasminogen activator inhibitor-1 function. AB - The adhesive glycoprotein vitronectin (VN) forms a function-stabilizing complex with plasminogen activator inhibitor-1 (PAI-1), the major fibrinolysis inhibitor in both plasma and vessel wall connective tissue. VN also interacts with two chain high molecular weight kininogen (HKa), particularly its His-Gly-Lys-rich domain 5, and both HKa and PAI-1 are antiadhesive factors that have been shown to compete for binding to VN. In this study the influence of HKa and domain 5 on the antifibrinolytic function of PAI-1 was investigated. In a purified system, HKa and particularly domain 5 inhibited the binding of PAI-1 to VN and promoted PAI-1 displacement from both isolated VN as well as subendothelial extracellular matrix associated VN. The sequence Gly(486)-Lys(502) of HKa domain 5 was identified as responsible for this inhibition. Although having no direct effect on PAI-1 activity itself, HKa domain 5 or the peptide Gly(486)-Lys(502) markedly destabilized the VN.PAI-1 complex interaction, resulting in a significant reduction of PAI-1 inhibitory function on plasminogen activators, resembling the effect of VN antibodies that prevent stabilization of PAI-1. Furthermore, high affinity fibrin binding of PAI-1 in the presence of VN as well as the VN dependent fibrin clot stabilization by the inhibitor were abrogated in the presence of the kininogen forms mentioned. Taken together, our data indicate that the peptide Gly(486)-Lys(502) derived from domain 5 of HKa serves to interfere with PAI-1 function. Based on these observations potential low molecular weight PAI-1 inhibitors could be designed for the use in therapeutic interventions against thromboembolic complications. PMID- 12082111 TI - Regulation of histone deacetylase 2 by protein kinase CK2. AB - Histone deacetylase 2 (HDAC2) is a member of a large family of enzymes that alter gene expression by catalyzing the removal of acetyl groups from core histones. Originally isolated as a transcriptional co-repressor, HDAC2 possesses extensive amino acid sequence homology to HDAC1 (the founding member and most extensively studied HDAC enzyme). Because of this high degree of sequence similarity between HDAC1 and HDAC2, coupled with the fact that the two always co-exist in the same complexes, it is difficult to assess whether different properties exist between these two proteins. We report here that HDAC2 is a phosphoprotein similar to HDAC1. In addition, like HDAC1, the phospho-acceptor sites in HDAC2 are located in the C-terminal portion of the protein. However, unlike HDAC1, which can be phosphorylated by protein kinase CK2, cAMP-dependent protein kinase, and protein kinase G, HDAC2 is phosphorylated uniquely by protein kinase CK2 in vitro. Studies using unfractionated cell extracts with CK2 inhibitors suggest that protein kinase CK2 is the major source of HDAC2 kinase. Finally, and perhaps most interesting, HDAC2 phosphorylation promotes enzymatic activity, selectively regulates complex formation, but has no effect on transcriptional repression. Together, our data indicate that like many HDACs, HDAC2 is regulated by post translational modification, particularly phosphorylation. Furthermore, we demonstrate for the first time that there are similarities and differences in the regulation of HDAC1 and HDAC2 by phosphorylation. PMID- 12082112 TI - Inactivation, complementation, and heterologous expression of encP, a novel bacterial phenylalanine ammonia-lyase gene. AB - The enzyme phenylalanine ammonia-lyase, which catalyzes the nonoxidative deamination of l-phenylalanine to trans-cinnamic acid, is ubiquitously distributed in plants. We now report its characterization for the first time in a bacterium. The phenylalanine ammonia-lyase homologous gene encP from the "Streptomyces maritimus" enterocin biosynthetic gene cluster was functionally characterized and shown to encode the first enzyme in the pathway to the enterocin polyketide synthase starter unit benzoyl-coenzyme A. The disruption of the encP gene completely inhibited the production of cinnamate and enterocin, whereas complementation of the mutant with benzoyl-coenzyme A pathway intermediates or with the wild-type gene encP restored the formation of the benzoate-primed polyketide antibiotic enterocin. Heterologous expression of the encP gene under the control of the ermE* promoter in Streptomyces coelicolor furthermore led to the production of cinnamic acid in the fermented cultures, confirming that the encP gene indeed encodes a novel bacterial phenylalanine ammonia-lyase. PMID- 12082113 TI - PEPT1 as a paradigm for membrane carriers that mediate electrogenic bidirectional transport of anionic, cationic, and neutral substrates. AB - The capability for electrogenic inward transport of substrates that carry different net charge is a phenomenon observed in a variety of membrane-solute transporters but is not yet understood. We employed the two-electrode voltage clamp technique combined with intracellular pH recordings and the giant patch technique to assess the selectivity for bidirectional transport and the underlying stoichiometries in proton to substrate flux coupling for electrogenic transfer of selected anionic, cationic, and neutral dipeptides by the intestinal peptide transporter PEPT1. Anionic dipeptides such as Gly-Asp and Asp-Gly are transported in their neutral and negatively charged forms with high and low affinities, respectively. The positive transport current obtained with monoanionic substrates results from the cotransport of two protons. Cationic dipeptides can be transported in neutral and positively charged form, resulting in an excess transport current as compared with neutral substrates. However, binding and transport of cationic dipeptides shows a pronounced selectivity for the position of charged side chains demonstrating that the binding domain of PEPT1 is asymmetric, both in its inward and outward facing conformation. The simultaneous presence of identically charged substrates on both membrane surfaces generates outward and, unexpectedly, enhanced inward transport currents probably by increasing the turnover rate. PMID- 12082114 TI - Intramolecular versus intermolecular disulfide bonds in prion proteins. AB - Prion protein (PrP) is the major component of the partially protease-resistant aggregate that accumulates in mammals with transmissible spongiform encephalopathies. The two cysteines of the scrapie form, PrP(Sc), were found to be in their oxidized (i.e. disulfide) form (Turk, E., Teplow, D. B., Hood, L. E., and Prusiner, S. B. (1988) Eur. J. Biochem. 176, 21-30); however, uncertainty remains as to whether the disulfide bonds are intra- or intermolecular. It is demonstrated here that the monomers of PrP(Sc) are not linked by intermolecular disulfide bonds. Furthermore, evidence is provided that PrP(Sc) can induce the conversion of the oxidized, disulfide-intact form of the monomeric cellular prion protein to its protease-resistant form without the temporary breakage and subsequent re-formation of the disulfide bonds in cell-free reactions. PMID- 12082115 TI - Thiazolidinedione activation of peroxisome proliferator-activated receptor gamma can enhance mitochondrial potential and promote cell survival. AB - Thiazolidinediones (TZDs) are widely used for treatment of type 2 diabetes mellitus. Peroxisome proliferator-activated receptor gamma (PPAR gamma) is the molecular target of TZDs and is believed to mediate the apoptotic effects of this class of drugs in a variety of cell types, including B and T lymphocytes. The finding that TZDs induce lymphocyte death has raised concerns regarding whether TZDs might further impair immune functions in diabetics. To address this issue, we investigated the roles of PPAR gamma and TZDs in lymphocyte survival. PPAR gamma was up-regulated upon T cell activation. As previously reported, PPAR gamma agonists induced T cell death in a dose-dependent manner. However, the concentrations of TZD needed to cause T cell death were above those needed to induce PPAR gamma-dependent transcription. Surprisingly, at concentrations that induce optimal transcriptional activation, TZD activation of PPAR gamma protected cells from apoptosis following growth factor withdrawal. The survival-enhancing effects depended on both the presence and activation of PPAR gamma. Measurements of mitochondrial potential revealed that PPAR gamma activation enhanced the ability of cells to maintain their mitochondrial potential. These data indicate that activation of PPAR gamma with TZDs can promote cell survival and suggest that PPAR gamma activation may potentially augment the immune responses of diabetic patients. PMID- 12082116 TI - Dithionite reduction kinetics of the dissimilatory copper-containing nitrite reductase of Alcalegenes xylosoxidans. The SO(2)(.-) radical binds to the substrate binding type 2 copper site before the type 2 copper is reduced. AB - We report here the first detailed study of the dithionite reduction kinetics of a copper-containing dissimilatory nitrite reductase (NiR). The reduction of the blue type 1 copper (T1Cu) center of NiR preparations that contained both type 1 and type 2 copper atoms, followed biphasic kinetics. In contrast, NiR that was deficient in type 2 copper (T2DNiR), followed monophasic kinetics with a second order rate constant (T2D)k = 3.06 x 10(6) m(-1) s(-1). In all cases the SO(2)(.-) radical rather than S(2)O(4)(2-) was the effective reductant. The observed kinetics were compatible with a reaction mechanism in which the T1Cu of the fully loaded protein is reduced both directly by dithionite and indirectly by the type 2 Cu (T2Cu) site via intramolecular electron transfer. Reduction kinetics of the T2Cu were consistent with SO(2)(.-) binding first to the T2Cu center and then transferring electrons (112 s(-1)) to reduce it. As SO(2)(.-) is a homologue of NO(2)(-), the NiR substrate, it is not unlikely that it binds to the catalytic T2Cu site. Effects on the catalytic activity of the enzyme using dithionite as a reducing agent are discussed. Reduction of the semireduced T1Cu(I)T2Cu(II) state followed either second-order kinetics with k(2) = 3.33 x 10(7) m(-1) s(-1) or first-order kinetics with 52.6 s(-1) < (T1red)k(1) < 112 s(-1). Values of formation constants of the T1Cu(II)T2Cu(II)-SO(2)(.-) and T1Cu(I)T2Cu(II)-SO(2)(. ) adducts showed that the redox state of T1Cu affected binding of SO(2)(.-) at the catalytic T2Cu center. Analysis of the kinetics required the development of a mathematical protocol that could be applied to a system with two intercommunicating sites but only one of which can be monitored. This novel protocol, reported for the first time, is of general application. PMID- 12082118 TI - Is there a toxicological advantage for non-hyperbolic kinetics in cytochrome P450 catalysis? Functional allostery from "distributive catalysis". AB - The cytochrome P450s (CYPs) are the major enzymatic detoxification and drug metabolism system. Recently, it has become clear that several CYP isoforms exhibit positive and negative homotropic cooperativity. However, the toxicological implications of allosteric kinetics have not been considered, nor understood. The allosteric kinetics are particularly enigmatic in several respects. In many cases, CYPs bioactivate substrates to more toxic products, thus making it difficult to rationalize a functional advantage for positive cooperativity. Also, CYPs exhibit cooperativity with many structurally diverse ligands, in marked contrast to the specificity observed with other allosteric systems. Here, kinetic simulations are used to compare the probabilistic time- and concentration-dependent integrated toxicity function during conversion of substrate to product for CYP models exhibiting Michaelis-Menten (non-cooperative) kinetics, positive cooperativity, or negative cooperativity. The results demonstrate that, at low substrate concentrations, the slower substrate turnover afforded by cooperative CYPs compared with Michaelis-Menten enzymes can be a significant toxicological advantage, when toxic thresholds exist. When present, the advantage results from enhanced "distribution" of toxin in two pools, substrate and product, for an extended period, thus minimizing the chance that either exceeds its toxic threshold. At intermediate concentrations, the allosteric kinetics can be a modest advantage or modest disadvantage, depending on the kinetic parameters. However, at high substrate concentrations associated with a high probability of toxicity, fast turnover is desirable, and this advantage is provided also by the cooperative enzymes. For the positive homotropic cooperativity, the allosteric kinetics minimize the probability of toxicity over the widest range of system parameters. Furthermore, this apparent functional cooperativity is achieved without specific molecular recognition that is the hallmark of "traditional" allostery. PMID- 12082117 TI - N-terminal tyrosine residues within the potassium channel Kir3 modulate GTPase activity of Galphai. AB - trkB activation results in tyrosine phosphorylation of N-terminal Kir3 residues, decreasing channel activation. To determine the mechanism of this effect, we reconstituted Kir3, trkB, and the mu opioid receptor in Xenopus oocytes. Activation of trkB by BDNF (brain-derived neurotrophic factor) accelerated Kir3 deactivation following termination of mu opioid receptor signaling. Similarly, overexpression of RGS4, a GTPase-activating protein (GAP), accelerated Kir3 deactivation. Blocking GTPase activity with GTPgammaS also prevented Kir3 deactivation, and the GTPgammaS effect was not reversed by BDNF treatment. These results suggest that BDNF treatment did not reduce Kir3 affinity for Gbetagamma but rather acted to accelerate GTPase activity, like RGS4. Tyrosine phosphatase inhibition by peroxyvanadate pretreatment reversibly mimicked the BDNF/trkB effect, indicating that tyrosine phosphorylation of Kir3 may have caused the GTPase acceleration. Tyrosine to phenylalanine substitution in the N-terminal domain of Kir3.4 blocked the BDNF effect, supporting the hypothesis that phosphorylation of these tyrosines was responsible. Like other GAPs, Kir3.4 contains a tyrosine-arginine-glutamine motif that is thought to function by interacting with G protein catalytic domains to facilitate GTP hydrolysis. These data suggest that the N-terminal tyrosine hydroxyls in Kir3 normally mask the GAP activity and that modification by phosphorylation or phenylalanine substitution reveals the GAP domain. Thus, BDNF activation of trkB could inhibit Kir3 by facilitating channel deactivation. PMID- 12082120 TI - Mitochondrial genome of the colorless green alga Polytomella parva: two linear DNA molecules with homologous inverted repeat Termini. AB - Most of the well-characterized mitochondrial genomes from diverse green algal lineages are circular mapping DNA molecules; however, Chlamydomonas reinhardtii has a linear 15.8 kb unit mitochondrial genome with 580 or 581 bp inverted repeat ends. In mitochondrial-enriched fractions prepared from Polytomella parva (=P. agilis), a colorless, naturally wall-less relative of C. reinhardtii, we have detected two linear mitochondrial DNA (mtDNA) components with sizes of 13.5 and 3.5 kb. Sequences spanning 97% and 86% of the 13.5- and 3.5-kb mtDNAs, respectively, reveal that these molecules contain long, at least 1.3 kb, homologous inverted repeat sequences at their termini. The 3.5-kb mtDNA has only one coding region (nad6), the functionality of which is supported by both the relative rate at which it has accumulated nonsynonymous nucleotide substitutions and its absence from the 13.5-kb mtDNA which encodes nine genes (i.e., large and small subunit rRNA [LSU and SSU rRNA] genes, one tRNA gene, and six protein coding genes). On the basis of DNA sequence data, we propose that a variant start codon, GTG, is utilized by the P. parva 13.5-kb mtDNA-encoded gene, nad5. Using the relative rate test with Chlamydomonas moewusii (=C. eugametos) as the outgroup, we conclude that the nonsynonymous nucleotide substitution rate in the mitochondrial protein-coding genes of P. parva is on an average about 3.3 times that of the C. reinhardtii counterparts. PMID- 12082119 TI - DNA sequence variation at a duplicated gene: excess of replacement polymorphism and extensive haplotype structure in the Drosophila melanogaster bicoid region. AB - The bicoid (bcd) gene of Drosophila has played an important role in understanding the system of developmental regulatory genes that controls segmentation in the fruit fly. Several studies in Drosophila and closely related insects suggest that bcd may be the result of a gene duplication in the Dipteran lineage. In addition, the presence of a large, conserved secondary structure in the 3' untranslated region (UTR) makes the bcd gene a good candidate for studying compensatory evolution and the relationship between RNA secondary structure and patterns of standing variation in natural populations. Despite these interesting aspects, a population-level analysis has until now not been performed on bcd. In this study, DNA sequence variation was examined for a 4-kb region of the bcd gene, including a portion of the 5' UTR, the entire coding region, and the 3' UTR, for 25 Drosophila melanogaster isofemale lines from Zimbabwe and one allele from D. simulans. Statistical tests revealed a significant excess of replacement polymorphisms in the D. melanogaster lineage that are clustered in two putative linker regions of the Bicoid protein. This result is consistent with a relaxation of selective constraints in these regions. In addition, we found a distinct haplotype structure and a significantly smaller number of haplotypes than predicted by the standard neutral model. It is unlikely that the haplotype structure is maintained by epistatic selection acting on the secondary structure in the 3' UTR or by the association of the bcd gene with polymorphic inversions. Instead, our two main observations, namely the occurrence of a haplotype structure and the excess of replacement polymorphisms, may indicate that the selective history of this gene is rather complex, involving both the relaxation of purifying selection in some parts of the protein and the action of positive selection in other parts of the gene region. PMID- 12082121 TI - Y chromosome evidence for Anglo-Saxon mass migration. AB - British history contains several periods of major cultural change. It remains controversial as to how much these periods coincided with substantial immigration from continental Europe, even for those that occurred most recently. In this study, we examine genetic data for evidence of male immigration at particular times into Central England and North Wales. To do this, we used 12 biallelic polymorphisms and six microsatellite markers to define high-resolution Y chromosome haplotypes in a sample of 313 males from seven towns located along an east-west transect from East Anglia to North Wales. The Central English towns were genetically very similar, whereas the two North Welsh towns differed significantly both from each other and from the Central English towns. When we compared our data with an additional 177 samples collected in Friesland and Norway, we found that the Central English and Frisian samples were statistically indistinguishable. Using novel population genetic models that incorporate both mass migration and continuous gene flow, we conclude that these striking patterns are best explained by a substantial migration of Anglo-Saxon Y chromosomes into Central England (contributing 50%-100% to the gene pool at that time) but not into North Wales. PMID- 12082122 TI - Ratios of radical to conservative amino acid replacement are affected by mutational and compositional factors and may not be indicative of positive Darwinian selection. AB - The ratio of radical to conservative amino acid replacements is frequently used to infer positive Darwinian selection. This method is based on the assumption that radical replacements are more likely than conservative replacements to improve the function of a protein. Therefore, if positive selection plays a major role in the evolution of a protein, one would expect the radical-conservative ratio to exceed the expectation under neutrality. Here, we investigate the possibility that factors unrelated to selection, i.e., transition-transversion ratio, codon usage, genetic code, and amino acid composition, influence the radical-conservative replacement ratio. All factors that have been studied were found to affect the radical-conservative replacement ratio. In particular, amino acid composition and transition-transversion ratio are shown to have the most profound effects. Because none of the studied factors had anything to do with selection (positive or otherwise) and also because all of them (singly or in combination) affected a measure that was supposed to be indicative of positive selection, we conclude that selectional inferences based on radical-conservative replacement ratios should be treated with suspicion. PMID- 12082123 TI - Isolation and characterization of the genomic region from Drosophila kuntzei containing the Adh and Adhr genes. AB - The nucleotide sequences of the Adh and Adhr genes of Drosophila kuntzei were derived from combined overlapping sequences of clones isolated from a genomic library and from cloned PCR and inverse-PCR fragments. Only a proximal promoter was detected upstream of the Adh gene, indicating that D. kuntzei Adh is regulated by a one-promoter system. Further upstream of the Adh structural gene, an adult enhancer region (AAE) was found that contains most of the regulatory sequences described for AAEs of other Drosophila species. Analysis of the ADH protein showed an amino acid change from valine to threonine in the active site at position 189 which is also found in D. funebris but is otherwise unique among Drosophila. This difference alone may be responsible for the very low ADH activity found in this species and may cause a difference in substrate usage pattern. Codon bias in Adh and Adhr was comparable and found to be very low compared with other species. Phylogenetic analysis showed that D. kuntzei is closest related to D. funebris and D. immigrans. The time of divergence between D. kuntzei and D. funebris was estimated to be 14.2-20.2 Myr and that between D. kuntzei-D. funebris and D. immigrans to be 30.8-44.0 Myr. An analysis of the genetic variation in the Adh gene and upstream sequences of four European strains showed that this gene was highly variable. Overall nucleotide diversity (pi) was 0.0139, which is two times higher than that in D. melanogaster. PMID- 12082124 TI - Identification and phylogenetic analysis of Drosophila melanogaster myosins. AB - Myosins constitute a superfamily of motor proteins that convert energy from ATP hydrolysis into mechanical movement along the actin filaments. Phylogenetic analysis currently places myosins into 17 classes based on class-specific features of their conserved motor domain. Traditionally, the myosins have been divided into two classes depending on whether they form monomers or dimers. The conventional myosin of muscle and nonmuscle cells forms class II myosins. They are complex molecules of four light chains bound to two heavy chains that form bipolar filaments via interactions between their coiled-coil tails (type II). Class I myosins are smaller monomeric myosins referred to as unconventional myosins. Now, at least 15 other classes of unconventional myosins are known. How many myosins are needed to ensure the proper development and function of eukaryotic organisms? Thus far, three types of myosins were found in budding yeast, six in the nematode Caenorhabditis elegans, and at least 12 in human. Here, we report on the identification and classification of Drosophila melanogaster myosins. Analysis of the Drosophila genome sequence identified 13 myosin genes. Phylogenetic analysis based on the sequence comparison of the myosin motor domains, as well as the presence of the class-specific domains, suggests that Drosophila myosins can be divided into nine major classes. Myosins belonging to previously described classes I, II, III, V, VI, and VII are present. Molecular and phylogenetic analysis indicates that the fruitfly genome contains at least five new myosins. Three of them fall into previously described myosin classes I, VII, and XV. Another myosin is a homolog of the mouse and human PDZ containing myosins, forming the recently defined class XVIII myosins. PDZ domains are named after the postsynaptic density, disc-large, ZO-1 proteins in which they were first described. The fifth myosin shows a unique domain composition and a low homology to any of the existing classes. We propose that this is classified when similar myosins are identified in other species. PMID- 12082125 TI - Rodent phylogeny and a timescale for the evolution of Glires: evidence from an extensive taxon sampling using three nuclear genes. AB - Rodentia is the largest order of placental mammals, with approximately 2,050 species divided into 28 families. It is also one of the most controversial with respect to its monophyly, relationships between families, and divergence dates. Here, we have analyzed and compared the performance of three nuclear genes (von Willebrand Factor, interphotoreceptor retinoid-binding protein, and Alpha 2B adrenergic receptor) for a large taxonomic sampling, covering the whole rodent and placental diversity. The phylogenetic results significantly support rodent monophyly, the association of Rodentia with Lagomorpha (the Glires clade), and a Glires + Euarchonta (Primates, Dermoptera, and Scandentia) clade. The resolution of relationships among rodents is also greatly improved. The currently recognized families are divided here into seven well-defined clades (Anomaluromorpha, Castoridae, Ctenohystrica, Geomyoidea, Gliridae, Myodonta, and Sciuroidea) that can be grouped into three major clades: Ctenohystrica, Gliridae + Sciuroidea, and a mouse-related clade (Anomaluromorpha, Castoridae + Geomyoidea, and Myodonta). Molecular datings based on these three genes suggest that the rodent radiation took place at the transition between Paleocene and Eocene. The divergence between rodents and lagomorphs is placed just at the K-T boundary and the first splits among placentals in the Late Cretaceous. Our results thus tend to reconcile molecular and morphological-paleontological insights. PMID- 12082126 TI - Phylogenetic and expression analysis of the glutamate-receptor-like gene family in Arabidopsis thaliana. AB - The ionotropic glutamate receptor (iGluR) gene family has been widely studied in animals and is determined to be important in excitatory neurotransmission and other neuronal processes. We have previously identified ionotropic glutamate receptor-like genes (GLRs) in Arabidopsis thaliana, an organism that lacks a nervous system. Upon the completion of the Arabidopsis genome sequencing project, a large family of GLR genes has been uncovered. A preliminary phylogenetic analysis divides the AtGLR gene family into three clades and is used as the basis for the recently established nomenclature for the AtGLR gene family. We performed a phylogenetic analysis with extensive annotations of the iGluR gene family, which includes all 20 Arabidopsis GLR genes, the entire iGluR family from rat (except NR3), and two prokaryotic iGluRs, Synechocystis GluR0 and Anabaena GluR. Our analysis supports the division of the AtGLR gene family into three clades and identifies potential functionally important amino acid residues that are conserved in both prokaryotic and eukaryotic iGluRs as well as those that are only conserved in AtGLRs. To begin to investigate whether the three AtGLR clades represent different functional classes, we performed the first comprehensive mRNA expression analysis of the entire AtGLR gene family. On the basis of RT-PCR, all AtGLRs are expressed genes. The three AtGLR clades do not show distinct clade specific organ expression patterns. All 20 AtGLR genes are expressed in the root. Among them, five of the nine clade-II genes are root-specific in 8-week-old Arabidopsis plants. PMID- 12082127 TI - Functional promiscuity of squirrel monkey growth hormone receptor toward both primate and nonprimate growth hormones. AB - Primate growth hormone (GH) has evolved rapidly, having undergone approximately 30% amino acid substitutions from the inferred ancestral eutherian sequence. Nevertheless, human growth hormone (hGH) is physiologically effective when administered to nonprimate mammals. In contrast, its functional counterpart, the human growth hormone receptor (hGHR), has evolved species specificity so that it responds only to Old World primate GHs. It has been proposed that this species specificity of the hGHR is largely caused by the Leu --> Arg change at position 43 after a prior His --> Asp change at position 171 of the GH. Sequence analyses supported this hypothesis and revealed that the transitional phase in the GH:GHR coevolution still persists in New World monkeys. For example, although the GH of the squirrel monkey has the His --> Asp substitution at position 171, residue 43 of its GHR is a Leu, the nonprimate residue. If the squirrel monkey truly represents an intermediate stage of GH:GHR coevolution, its GHR should respond to both hGH and nonprimate GH. Also, if the emergence of species specificity was a result of the selection for a more efficient GH:GHR interaction, then changing residue 43 of the squirrel monkey growth hormone receptor (smGHR) to Arg should increase its binding affinity toward higher primate GH. To test these hypotheses, we performed protein-binding assays between the smGHR and both human and rat GHs, using the surface plasmon resonance methodology. Furthermore, the effects of reciprocal mutations at position 43 of human and squirrel monkey GHRs are measured for their binding affinities toward human and squirrel monkey GHs. The results from the binding kinetic assays clearly demonstrate that the smGHR is in the intermediate state of the evolution of species specificity. Interestingly, the altered residue Arg at position 43 of the smGHR does not lead to an increased binding affinity. The implications of these results on the evolution of the GH:GHR interaction and on functional evolution are discussed. PMID- 12082128 TI - Adaptive evolution of the IgA hinge region in primates. AB - IgA is a major component that prevents the penetration of pathogenic bacteria into mucosal surfaces. The IgA antibody is cleaved at the IgA hinge region with high specificity by IgA-specific proteases produced by several pathogenic bacteria. We conducted a genomic sequence analysis of the IgA genes of a wide spectrum of primates, including the first intron and second exon, which consist of the hinge region and the CH2 domain, to find evidence of positive selection. Because the hinge region is quite small, we combined the largest collection of sequences that could be clearly aligned and evaluated the total numbers of synonymous and nonsynonymous substitutions on the phylogenetic tree. The nonsynonymous to synonymous substitution ratio (d(N)/d(S) test) showed that hominoids, Old World monkeys, and New World monkeys have d(N)/d(S) ratios of 5.4, 6.3, and 4.2, respectively. Fisher's exact probability tests showed statistical significance for the Old World monkey. Because the substitution rates of the flanking sequences are more or less similar to the synonymous rates of the hinge region, these high values of d(N)/d(S) should be the result of positive selection at the hinge region. Combining the high sequence variability in each population and the highly accelerated nonsynonymous substitution rates in the hinge region, we conclude that this unusual IgA evolution is a molecular evidence of adaptive evolution possibly caused by the host-parasite relationship. PMID- 12082130 TI - Evolution of transcription factor binding sites in Mammalian gene regulatory regions: conservation and turnover. AB - Comparisons between human and rodent DNA sequences are widely used for the identification of regulatory regions (phylogenetic footprinting), and the importance of such intergenomic comparisons for promoter annotation is expanding. The efficacy of such comparisons for the identification of functional regulatory elements hinges on the evolutionary dynamics of promoter sequences. Although it is widely appreciated that conservation of sequence motifs may provide a suggestion of function, it is not known as to what proportion of the functional binding sites in humans is conserved in distant species. In this report, we present an analysis of the evolutionary dynamics of transcription factor binding sites whose function had been experimentally verified in promoters of 51 human genes and compare their sequence to homologous sequences in other primate species and rodents. Our results show that there is extensive divergence within the nucleotide sequence of transcription factor binding sites. Using direct experimental data from functional studies in both human and rodents for 20 of the regulatory regions, we estimate that 32%-40% of the human functional sites are not functional in rodents. This is evidence that there is widespread turnover of transcription factor binding sites. These results have important implications for the efficacy of phylogenetic footprinting and the interpretation of the pattern of evolution in regulatory sequences. PMID- 12082129 TI - Contrasting rates of mitochondrial molecular evolution in parasitic Diptera and Hymenoptera. AB - We investigated the putative association between the parasitic lifestyle and an accelerated rate of mt genetic divergence, compositional bias, and gene rearrangement, employing a range of parasitic and nonparasitic Diptera and Hymenoptera. Sequences were obtained for the cox1, cox2, 16S, 28S genes, the regions between the cox2 and atp8 genes, and between the nad3 and nad5 genes. Relative rate tests indicated generally that the parasitic lifestyle was not associated with an increased rate of genetic divergence in the Diptera but reaffirmed that it was in the Hymenoptera. Similarly, a departure from compositional stationarity was not associated with parasitic Diptera but was in parasitic Hymenoptera. Finally, mitochondrial (mt) gene rearrangements were not observed in any of the dipteran species examined. The results indicate that these genetic phenomena are not accelerated in parasitic Diptera compared with nonparasitic Diptera. A possible explanation for the differences in the rate of mt molecular evolution in parasitic Diptera and Hymenoptera is the extraordinary level of radiation that has occurred within the parasitic Hymenoptera but not in any of the dipteran parasitic lineages. If speciation events in the parasitic Hymenoptera are associated with founder events, a faster rate of molecular evolution is expected. Alternatively, biological differences between endoparasitic Hymenoptera and endoparasitic Diptera may also account for the differences observed in molecular evolution. PMID- 12082131 TI - Recombination or mutational hot spots in human mtDNA? AB - Awadalla, Eyre-Walker, and Maynard Smith (1999) recently argued that there might be recombination in human mitochondrial DNA (mtDNA). Their claim was based on their observation of decaying linkage disequilibrium (LD) as a function of physical distance. Their study was much criticized, and follow-up studies have failed to find any evidence for recombination. We argue that the criticisms levied, even if correct, could not possibly explain the findings of Awadalla, Eyre-Walker, and Maynard Smith (1999). Nonetheless, the test proposed by Awadalla, Eyre-Walker, and Maynard Smith (1999 ) is not robust because recombination is not the only explanation for decay of LD. We show that such a pattern can be caused by mutational hot spots as well. However, a closer look at the data suggests that the pattern observed was not caused by mutational hot spots but rather by chance. Thus, there appears to be no evidence for recombination in the mtDNA polymorphism data. In conclusion, we discuss the possibility of detecting recombination in mtDNA and the implications of its existence. PMID- 12082132 TI - Evolutionary relationships of conserved cysteine-rich motifs in adhesive molecules of malaria parasites. AB - Malaria parasites invade erythrocytes in a process mediated by a series of molecular interactions. Invasion of human erythrocytes by Plasmodium vivax is dependent upon the presence of a single receptor, but P. falciparum, as well as some other species, exhibits the ability to utilize multiple alternative invasion pathways. Conserved cysteine-rich domains play important roles at critical times during this invasion process and at other stages in the life cycle of malaria parasites. Duffy-binding-like (DBL) domains, expressed as a part of the erythrocyte-binding proteins (DBL-EBP), are such essential cysteine-rich ligands that recognize specific host cell surface receptors. DBL-EBP, which are products of the erythrocyte-binding-like (ebl) gene family, act as critical determinants of erythrocyte specificity and are the best-defined ligands from invasive stages of malaria parasites. The ebl genes include the P. falciparum erythrocyte-binding antigen-175 (EBA-175) and P. vivax Duffy-binding protein. DBL domains also mediate cytoadherence as a part of the variant erythrocytic membrane protein-1 (PfEMP-1) antigens expressed from var genes on the surface of P. falciparum infected erythrocytes. A paralogue of the ebl family is the malarial ligand MAEBL, which has a chimeric structure where the DBL domain is functionally replaced with a distinct cysteine-rich erythrocyte-binding domain with similarity to the apical membrane antigen-1 (AMA-1) ligand domain. The Plasmodium AMA-1 ligand domain, which encompasses the extracellular cysteine domains 1 and 2 and is well conserved in a Toxoplasma gondii AMA-1, has erythrocyte-binding activity distinct from that of MAEBL. These important families of Plasmodium molecules (DBL-EBP, PfEMP-1, MAEBL, AMA-1) are interrelated through the MAEBL. Because MAEBL and the other ebl products have the characteristics expected of homologous ligands involved in equivalent alternative invasion pathways to each other, we sought to better understand their roles during invasion by determining their relative origins in the Plasmodium genome. An analysis of their multiple cysteine rich domains permitted a unique insight into the evolutionary development of PLASMODIUM: Our data indicate that maebl, ama-1, and ebl genes have ancient origins which predate Plasmodium speciation. The maebl evolved as a single locus, including its unique chimeric structure, in each Plasmodium species, in parallel with the ama-1 and the ebl genes families. The ancient character of maebl, along with its different expression characteristics suggests that MAEBL is unique and does not play an alternative role in invasion to ebl products such as EBA-175. The multiple P. falciparum ebl paralogues that express DBL domains, which have occurred by duplication and diversification, potentially do provide multiple functionally equivalent ligands to EBA-175 for alternative invasion pathways. PMID- 12082133 TI - Searching for evidence of positive selection in the human genome using patterns of microsatellite variability. AB - Both natural selection and nonequilibrium population-level processes can lead to a skew in the frequency distribution of polymorphisms. Population-level processes are expected to affect all loci in a roughly equal fashion, whereas selection will affect only some regions of the genome. We conducted a sliding-window analysis of the frequency distribution of microsatellite polymorphisms across the human genome to identify regions that may be under positive selection. The analysis was based on a published data set of 5,257 mapped microsatellites in individuals of European ancestry. Observed and expected numbers of alleles were compared under a stepwise mutation model (SMM) using analytical formulae. Observed and expected heterozygosities were compared under a SMM using coalescent simulations. The two sets of analyses gave similar results. Approximately one fourth of all loci showed a significant deficit of heterozygosity, consistent with a recent population expansion. Forty-three windows were identified with extreme skews in the frequency distribution of polymorphisms (in the direction of a deficit of heterozygosity, given the number of alleles). If these extreme windows are tracking selection at linked sites, theory predicts that they should be more common in regions of the genome with less recombination. We tested this prediction by comparing recombination rates in these extreme windows and in other regions of the genome and found that extreme windows had a significantly lower recombination rate than the genomic average. The proportion of extreme windows was significantly higher on the X chromosome than on the autosomes. Moreover, all the windows with extreme skews on the X chromosome were found in two clusters near the centromere; both these clusters exhibit markedly reduced recombination rates. These analyses point to regions of the genome that may recently have been subject to positive selection. These results also suggest that the effects of positive selection may be more pronounced on the X chromosome than on the autosomes in humans. PMID- 12082134 TI - Evolution of genome size in Drosophila. is the invader's genome being invaded by transposable elements? AB - Genome size varies considerably between species, and transposable elements (TEs) are known to play an important role in this variability. However, it is far from clear whether TEs are involved in genome size differences between populations within a given species. We show here that in Drosophila melanogaster and Drosophila simulans the size of the genome varies among populations and is correlated with the TE copy number on the chromosome arms. The TEs embedded within the heterochromatin do not seem to be involved directly in this phenomenon, although they may contribute to differences in genome size. Furthermore, genome size and TE content variations parallel the worldwide colonization of D. melanogaster species. No such relationship exists for the more recently dispersed D. simulans species, which indicates that a quantitative increase in the TEs in local populations and fly migration are sufficient to account for the increase in genome size, with no need for an adaptation hypothesis. PMID- 12082135 TI - The evolution of the heat-shock protein GroEL from Buchnera, the primary endosymbiont of aphids, is governed by positive selection. AB - The heat-shock protein GroEL is a double-ring-structured chaperonin that assists the folding of many newly synthesized proteins in Escherichia coli and the refolding in vitro, with the cochaperonin GroES, of conformationally damaged proteins. This protein is constitutively overexpressed in the primary symbiotic bacteria of many insects, constituting approximately 10% of the total protein in Buchnera, the primary endosymbiont of aphids. In the present study, we perform a maximum likelihood (ML) analysis to unveil the selective constraints in GroEL. In addition, we apply a new statistical approach to determine the patterns of evolution in this highly interesting protein. The main conclusion derived from our analysis is that GroEL has suffered an accelerated rate of amino acid substitution upon the symbiotic integration of Buchnera into the aphids. It is most interesting that the ML analysis of codon substitutions in the different branches of the phylogenetic tree strongly supports the action of positive selection in the different lineages of BUCHNERA: Additionally, the new sliding window analysis of the complete groEL sequence reveals different regions of the molecule under the action of positive selection, mainly located in the apical domain, that are important for both peptide and GroES binding. PMID- 12082136 TI - Bayesian model adequacy and choice in phylogenetics. AB - Bayesian inference is becoming a common statistical approach to phylogenetic estimation because, among other reasons, it allows for rapid analysis of large data sets with complex evolutionary models. Conveniently, Bayesian phylogenetic methods use currently available stochastic models of sequence evolution. However, as with other model-based approaches, the results of Bayesian inference are conditional on the assumed model of evolution: inadequate models (models that poorly fit the data) may result in erroneous inferences. In this article, I present a Bayesian phylogenetic method that evaluates the adequacy of evolutionary models using posterior predictive distributions. By evaluating a model's posterior predictive performance, an adequate model can be selected for a Bayesian phylogenetic study. Although I present a single test statistic that assesses the overall (global) performance of a phylogenetic model, a variety of test statistics can be tailored to evaluate specific features (local performance) of evolutionary models to identify sources failure. The method presented here, unlike the likelihood-ratio test and parametric bootstrap, accounts for uncertainty in the phylogeny and model parameters. PMID- 12082137 TI - Integrating genomics, bioinformatics, and classical genetics to study the effects of recombination on genome evolution. AB - This study presents compelling evidence that recombination significantly increases the silent GC content of a genome in a selectively neutral manner, resulting in a highly significant positive correlation between recombination and "GC3s" in the yeast Saccharomyces cerevisiae. Neither selection nor mutation can explain this relationship. A highly significant GC-biased mismatch repair system is documented for the first time in any member of the Kingdom Fungi. Much of the variation in the GC3s within yeast appears to result from GC-biased gene conversion. Evidence suggests that GC-biased mismatch repair exists in numerous organisms spanning six kingdoms. This transkingdom GC mismatch repair bias may have evolved in response to a ubiquitous AT mutational bias. A significant positive correlation between recombination and GC content is found in many of these same organisms, suggesting that the processes influencing the evolution of the yeast genome may be a general phenomenon. Nonrecombining regions of the genome and nonrecombining genomes would not be subject to this type of molecular drive. It is suggested that the low GC content characteristic of many nonrecombining genomes may be the result of three processes (1) a prevailing AT mutational bias, (2) random fixation of the most common types of mutation, and (3) the absence of the GC-biased gene conversion which, in recombining organisms, permits the reversal of the most common types of mutation. A model is proposed to explain the observation that introns, intergenic regions, and pseudogenes typically have lower GC content than the silent sites of corresponding open reading frames. This model is based on the observation that the greater the heterology between two sequences, the less likely it is that recombination will occur between them. According to this "Constraint" hypothesis, the formation and propagation of heteroduplex DNA is expected to occur, on average, more frequently within conserved coding and regulatory regions of the genome. In organisms possessing GC-biased mismatch repair, this would enhance the GC content of these regions through biased gene conversion. These findings have a number of important implications for the way we view genome evolution and suggest a new model for the evolution of sex. PMID- 12082138 TI - Rates and patterns of microsatellite mutations in pink salmon. PMID- 12082139 TI - Phylogenetic analysis indicates multiple origins of chloroplast glyceraldehyde-3 phosphate dehydrogenase genes in dinoflagellates. PMID- 12082140 TI - The dystrophin-associated protein complex. PMID- 12082143 TI - Elastic fibres. AB - Elastic fibres are essential extracellular matrix macromolecules comprising an elastin core surrounded by a mantle of fibrillin-rich microfibrils. They endow connective tissues such as blood vessels, lungs and skin with the critical properties of elasticity and resilience. The biology of elastic fibres is complex because they have multiple components, a tightly regulated developmental deposition, a multi-step hierarchical assembly and unique biomechanical functions. However, their molecular complexity is at last being unravelled by progress in identifying interactions between component molecules, ultrastructural analyses and studies of informative mouse models. PMID- 12082142 TI - Chaperoning signaling pathways: molecular chaperones as stress-sensing 'heat shock' proteins. AB - Heat shock proteins interact with multiple key components of signaling pathways that regulate growth and development. The molecular relationships between heat shock proteins, various signaling proteins and partner proteins appear to be critical for the normal function of signal transduction pathways. The relative levels of these proteins may be important, as too little or too much Hsp70 or Hsp90 can result in aberrant growth control, developmental malformations and cell death. Although the functions of heat shock proteins as molecular chaperones have been well characterized, their complementary role as a 'stress-induced' proteins to monitor changes and alter the biochemical environment of the cell remains elusive. Genetic and molecular interactions between heat shock proteins, their co chaperones and components of signaling pathways suggest that crosstalk between these proteins can regulate proliferation and development by preventing or enhancing cell growth and cell death as the levels of heat shock proteins vary in response to environmental stress or disease. PMID- 12082144 TI - Prolonged arrest of mammalian cells at the G1/S boundary results in permanent S phase stasis. AB - Mammalian cells in culture normally enter a state of quiescence during G1 following suppression of cell cycle progression by senescence, contact inhibition or terminal differentiation signals. We find that mammalian fibroblasts enter cell cycle stasis at the onset of S phase upon release from prolonged arrest with the inhibitors of DNA replication, hydroxyurea or aphidicolin. During arrest typical S phase markers remain present, and G0/G1 inhibitory signals such as p21(WAF1) and p27 are absent. Cell cycle stasis occurs in T-antigen transformed cells, indicating that p53 and pRB inhibitory circuits are not involved. While no DNA replication is evident in arrested cells, nuclei isolated from these cells retain measurable competence for in vitro replication. MCM proteins are required to license replication origins, and are put in place in nuclei in G1 and excluded from chromatin by the end of replication to prevent rereplication of the genome. Strikingly, MCM proteins are strongly depleted from chromatin during prolonged S phase arrest, and their loss may underlie the observed cell cycle arrest. S phase stasis may thus be a 'trap' in which cells otherwise competent for S phase have lost a key component required for replication and thus can neither go forward nor retreat to G1 status. PMID- 12082145 TI - Genome restructuring in rye affects the expression, organization and disposition of homologous rDNA loci. AB - The standard rye cultivar 'Imperial' and a structural variant carrying an intact 1R chromosome and two telocentric 1R chromosomes (short and long arms) were used to investigate expression patterns of homologous rDNA loci, and the influence of chromosome structural change on their interphase organisation and relative disposition. Sequential silver staining and in situ hybridization with the rDNA probe pTa71, established a correspondence between the expression and organization patterns of rDNA domains in metaphase and interphase cells. In most cells of the cultivar Imperial, nucleolar organizer region (NOR) silver staining on metaphase chromosomes with equivalent numbers of rDNA genes revealed a size heteromorphism between homologous rDNA loci, resulting from their differential expression. NOR heteromorphism in the structural variant line was significantly reduced. The preferential activity of one NOR over its homologue was found to be random within cells and independent of parental origin. Nucleotypic modifications mediated by changes in the 1R chromosome structure include increased proximity between homologous rDNA loci in interphase, and an increase in the frequency of cells with intra-nucleolar ribosomal condensed chromatin. These results seem to indicate a 'sequence recognition' process for the regulation of homologous loci. PMID- 12082146 TI - The dynamic localisation of the Drosophila APC/C: evidence for the existence of multiple complexes that perform distinct functions and are differentially localised. AB - In Drosophila cells, the destruction of cyclin B is spatially regulated. In cellularised embryos, cyclin B is initially degraded on the mitotic spindle and is then degraded in the cytoplasm. In syncytial embryos, only the spindle associated cyclin B is degraded at the end of mitosis. The anaphase promoting complex/cyclosome (APC/C) targets cyclin B for destruction, but its subcellular localisation remains controversial. We constructed GFP fusions of two core APC/C subunits, Cdc16 and Cdc27. These fusion proteins were incorporated into the endogenous APC/C and were largely localised in the cytoplasm during interphase in living syncytial embryos. Both fusion proteins rapidly accumulated in the nucleus prior to nuclear envelope breakdown but only weakly associated with mitotic spindles throughout mitosis. Thus, the global activation of a spatially restricted APC/C cannot explain the spatially regulated destruction of cyclin B. Instead, different subpopulations of the APC/C must be activated at different times to degrade cyclin B. Surprisingly, we noticed that GFP-Cdc27 associated with mitotic chromosomes, whereas GFP-Cdc16 did not. Moreover, reducing the levels of Cdc16 or Cdc27 by >90% in tissue culture cells led to a transient mitotic arrest that was both biochemically and morphologically distinct. Taken together, our results raise the intriguing possibility that there could be multiple forms of the APC/C that are differentially localised and perform distinct functions. PMID- 12082147 TI - Functional consequence of targeting protein kinase B/Akt to GLUT4 vesicles. AB - We have investigated the role of protein kinase B (Akt) in the insulin-stimulated translocation of vesicles containing the insulin-responsive isoform of glucose transporter (GLUT4) to the plasma membrane of adipocytes. Previous reports have suggested that protein kinase B can bind to intracellular GLUT4 vesicles in an insulin-dependent manner, but the functional consequence of this translocation is not known. In this study we have artificially targeted constitutively active and kinase-inactive mutants of protein kinase B to intracellular GLUT4 vesicles by fusing them with the N-terminus of GLUT4 itself. We examined the effect of these mutants on the insulin-dependent translocation of the insulin-responsive amino peptidase IRAP (a bona fide GLUT4-vesicle-resident protein). A kinase-inactive protein kinase B targeted to GLUT4 vesicles was an extremely effective dominant negative inhibitor of insulin-stimulated IRAP translocation to the plasma membrane. By contrast, a kinase-inactive protein kinase B expressed in the cytoplasm did not have an effect. The results suggest that protein kinase B has an important functional role at, or in the vicinity of, GLUT4 vesicles in the insulin-dependent translocation of those vesicles to the plasma membrane of adipocytes. PMID- 12082148 TI - A conserved C-terminal domain of EFA6-family ARF6-guanine nucleotide exchange factors induces lengthening of microvilli-like membrane protrusions. AB - We recently reported the identification of EFA6 (exchange factor for ARF6), a brain-specific Sec7-domain-containing guanine nucleotide exchange factor that works specifically on ARF6. Here, we have characterized the product of a broadly expressed gene encoding a novel 1056 amino-acid protein that we have named EFA6B. We show that EFA6B, which contains a Sec7 domain that is highly homologous to EFA6, works as an ARF6-specific guanine exchange factor in vitro. Like EFA6, which will be referred to as EFA6A from now on, EFA6B is involved in membrane recycling and colocalizes with ARF6 in actin-rich membrane ruffles and microvilli like protrusions on the dorsal cell surface in transfected baby hamster kidney cells. Strikingly, homology between EFA6A and EFA6B is not limited to the Sec7 domain but extends to an adjacent pleckstrin homology (PH) domain and a approximately 150 amino-acid C-terminal region containing a predicted coiled coil motif. Association of EFA6A with membrane ruffles and microvilli-like structures depends on the PH domain, which probably interacts with phosphatidylinositol 4,5 biphosphate. Moreover, we show that overexpression of the PH domain/C-terminal region of EFA6A or EFA6B in the absence of the Sec7 domain promotes lengthening of dorsal microvillar protrusions. This morphological change requires the integrity of the coiled-coil motif. Lastly, database analysis reveals that the EFA6-family comprises at least four members in humans and is conserved in multicellular organisms throughout evolution. Our results suggest that EFA6 family guanine exchange factors are modular proteins that work through the coordinated action of the catalytic Sec7 domain to promote ARF6 activation, through the PH domain to regulate association with specific subdomains of the plasma membrane and through the C-terminal region to control actin cytoskeletal reorganization. PMID- 12082149 TI - A novel linker histone-like protein is associated with cytoplasmic filaments in Caenorhabditis elegans. AB - The histone H1 complement of Caenorhabditis elegans contains a single unusual protein, H1.X. Although H1.X possesses the globular domain and the canonical three-domain structure of linker histones, the amino acid composition of H1.X is distinctly different from conventional linker histones in both terminal domains. We have characterized H1.X in C. elegans by antibody labeling, green fluorescent protein fusion protein expression and RNA interference. Unlike normal linker histones, H1.X is a cytoplasmic as well as a nuclear protein and is not associated with chromosomes. H1.X is most prominently expressed in the marginal cells of the pharynx and is associated with a peculiar cytoplasmic cytoskeletal structure therein, the tonofilaments. Additionally H1.X::GFP is expressed in the cytoplasm of body and vulva muscle cells, neurons, excretory cells and in the nucleoli of embryonic blastomeres and adult gut cells. RNA interference with H1.X results in uncoordinated and egg laying defective animals, as well as in a longitudinally enlarged pharynx. These phenotypes indicate a cytoplasmic role of H1.X in muscle growth and muscle function. PMID- 12082150 TI - Dynamics of the vacuolar H(+)-ATPase in the contractile vacuole complex and the endosomal pathway of Dictyostelium cells. AB - The vacuolar H(+)-ATPase (V-ATPase) is a multi-subunit enzyme that plays important roles in eukaryotic cells. In Dictyostelium, it is found primarily in membranes of the contractile vacuole complex, where it energizes fluid accumulation by this osmoregulatory organelle and also in membranes of endolysosomes, where it serves to acidify the endosomal lumen. In the present study, a fusion was created between vatM, the gene encoding the 100 kDa transmembrane subunit of the V-ATPase, and the gene encoding Green Fluorescent Protein (GFP). When expressed in Dictyostelium cells, this fusion protein, VatM GFP, was correctly targeted to contractile vacuole and endolysosomal membranes and was competent to direct assembly of the V-ATPase enzyme complex. Protease treatment of isolated endosomes indicated that the GFP moiety, located on the C terminus of VatM, was exposed to the cytoplasmic side of the endosomal membrane rather than to the lumenal side. VatM-GFP labeling of the contractile vacuole complex revealed clearly the dynamics of this pleiomorphic vesiculotubular organelle. VatM-GFP labeling of endosomes allowed direct visualization of the trafficking of vacuolar proton pumps in this pathway, which appeared to be entirely independent from the contractile vacuole membrane system. In cells whose endosomes were pre-labeled with TRITC-dextran and then fed yeast particles, VatM GFP was delivered to newly formed yeast phagosomes with the same time course as TRITC-dextran, consistent with transfer via a direct fusion of endosomes with phagosomes. Several minutes were required before the intensity of the VatM-GFP labeling of new phagosomes reached the level observed in older phagosomes, suggesting that this fusion process was progressive and continuous. VatM-GFP was retrieved from the phagosome membrane prior to exocytosis of the indigestible remnants of the yeast particle. These data suggest that vacuolar proton pumps are recycled by fusion of advanced with newly formed endosomes. PMID- 12082151 TI - The cellular fate of mutant rhodopsin: quality control, degradation and aggresome formation. AB - Mutations in the photopigment rhodopsin are the major cause of autosomal dominant retinitis pigmentosa. The majority of mutations in rhodopsin lead to misfolding of the protein. Through the detailed examination of P23H and K296E mutant opsin processing in COS-7 cells, we have shown that the mutant protein does not accumulate in the Golgi, as previously thought, instead it forms aggregates that have many of the characteristic features of an aggresome. The aggregates form close to the centrosome and lead to the dispersal of the Golgi apparatus. Furthermore, these aggregates are ubiquitinated, recruit cellular chaperones and disrupt the intermediate filament network. Mutant opsin expression can disrupt the processing of normal opsin, as co-transfection revealed that the wild-type protein is recruited to mutant opsin aggregates. The degradation of mutant opsin is dependent on the proteasome machinery. Unlike the situation with DeltaF508 CFTR, proteasome inhibition does not lead to a marked increase in aggresome formation but increases the retention of the protein within the ER, suggesting that the proteasome is required for the efficient retrotranslocation of the mutant protein. Inhibition of N-linked glycosylation with tunicamycin leads to the selective retention of the mutant protein within the ER and increases the steady state level of mutant opsin. Glycosylation, however, has no influence on the biogenesis and targeting of wild-type opsin in cultured cells. This demonstrates that N-linked glycosylation is required for ER-associated degradation of the mutant protein but is not essential for the quality control of opsin folding. The addition of 9-cis-retinal to the media increased the amount of P23H, but not K296E, that was soluble and reached the plasma membrane. These data show that rhodopsin autosomal dominant retinitis pigmentosa is similar to many other neurodegenerative diseases in which the formation of intracellular protein aggregates is central to disease pathogenesis, and they suggest a mechanism for disease dominance. PMID- 12082152 TI - Localization and possible role of two different alpha v beta 3 integrin conformations in resting and resorbing osteoclasts. AB - Integrins are membrane receptors that mediate interactions between cells and the extracellular matrix. We recently showed that the osteoclast integrin alpha(v)beta(3) exists in two different conformations, so-called 'basal' and 'activated', with each exhibiting a distinct function. In this study we demonstrate that, in non-resorbing osteoclasts, the 'activated' form of alpha(v)beta(3) accumulates in the motile areas of the plasma membrane. During bone resorption this conformation is prevalent in the ruffled membrane, whereas the 'basal' form of alpha(v)beta(3) is also present in the sealing zone. Moreover, hepatocyte growth factor (HGF) and macrophage colony stimulating factor (M-CSF), two molecules involved in osteoclastogenesis and osteoclast survival, modulate alpha(v)beta(3) conformation in vitro. Preincubation with HGF or M-CSF induces a shift of conformation of alpha(v)beta(3) in primary human osteoclasts (OCs) and in the osteoclast-like cell line (GCT 23). Activated integrin promotes osteoclast migration to the alpha(v)beta(3) ligand osteopontin and enhances bone resorption. Thus, HGF and M-CSF modulate the alpha(v)beta(3) conformational states required for osteoclast polarization and resorption. The capacity of growth factors to alter the affinity of alpha(v)beta(3) toward its ligands offers a potential explanation for the diverse responses of osteoclasts to the same ligand. PMID- 12082153 TI - Differential regulation of maternal and paternal chromosome condensation in mitotic zygotes. AB - A-kinase anchoring protein AKAP95 is implicated in somatic mitotic chromosome condensation by recruiting the condensin complex. Here, we report a differential regulation of condensation of maternal and paternal chromosomes mediated by AKAP95 in mitotic mouse zygotes. AKAP95 is synthesized upon oocyte activation, targeted to the female pronucleus and specifically associates with maternal chromosomes at mitosis. AKAP95 mRNA is highly restricted to the vicinity of the meiotic spindle in metaphase II oocytes. In vivo displacement of endogenous AKAP95 in female pronuclei by microinjection of competitor peptides and rescue experiments show that AKPA95 is required for recruitment of the mCAP-D2 condensin subunit to, and condensation of, maternal chromosomes. In contrast, AKAP95 is dispensable for mCAP-D2 recruitment to, and condensation of, paternal chromosomes. Our results indicate that at first embryonic mitosis, paternal chromosomes target condensins and condense independently of AKAP95, whereas maternal chromosomes require AKAP95 for condensin recruitment and condensation. We propose a concept whereby condensation of chromosomes in gametes, zygotes and somatic cells involves related but distinct mechanisms. PMID- 12082154 TI - Role of sulfated O-linked glycoproteins in zymogen granule formation. AB - Packaging of proteins into regulated secretory granules is mediated by the mildly acidic pH of the trans Golgi network and immature secretory granules. This need for an acidic pH indicates that ionic interactions are important. The mouse pancreatic acinar cell contains four major sulfated glycoproteins, including the zymogen granule structural component Muclin. I tested the hypothesis that sulfation and the O-linked glycosylation to which the sulfates are attached are required for normal formation of zymogen granules in the exocrine pancreas. Post translational processing was perturbed with two chemicals: sodium chlorate was used to inhibit sulfation and benzyl-N-acetyl-alpha-galactosaminide was used to inhibit O-linked oligosaccharide elongation. Both chemicals resulted in the accumulation in the Golgi region of the cell of large vacuoles that appear to be immature secretory granules, and the effect was much more extensive with benzyl-N acetyl-alpha-galactosaminide than chlorate. Both chemical treatments inhibited basal secretion at prolonged chase times, and again benzyl-N-acetyl-alpha galactosaminide had a greater effect than chlorate. In addition, benzyl-N-acetyl alpha-galactosaminide, but not chlorate, totally inhibited stimulated secretion of newly synthesized proteins. These data provide evidence for a role of sulfated O-linked glycoproteins in protein condensation and maturation of zymogen granules. Under maximal inhibition of O-linked oligosaccharide biosynthesis, anterograde post-Golgi traffic in the regulated pathway is almost totally shut down, demonstrating the importance of these post-translational modifications in progression of secretory proteins through the regulated pathway and normal granule formation in the pancreatic acinar cell. PMID- 12082155 TI - Membrane ruffling and macropinocytosis in A431 cells require cholesterol. AB - Cholesterol is important for the formation of caveolea and deeply invaginated clathrin-coated pits. We have now investigated whether formation of macropinosomes is dependent on the presence of cholesterol in the plasma membrane. Macropinocytosis in A431 cells was induced by the phorbol ester 12-O tetradecanoylphorbol 13-acetate, a potent activator of protein kinase C (PKC). When cells were pretreated with methyl-beta-cyclodextrin to extract cholesterol, the phorbol ester was unable to induce the increased endocytosis of ricin otherwise seen, although PKC could still be activated. Electron microscopy revealed that extraction of cholesterol inhibited the formation of membrane ruffles and macropinosomes at the plasma membrane. Furthermore, cholesterol depletion inhibited the phorbol ester-induced reorganization of filamentous actin at the cell periphery, a prerequisite for the formation of membrane ruffles that close into macropinosomes. Under normal conditions the small GTPase Rac1 is activated by the phorbol ester and subsequently localized to the plasma membrane, where it induces the reorganization of actin filaments required for formation of membrane ruffles. Cholesterol depletion did not inhibit the activation of Rac1. However, confocal microscopy showed that extraction of cholesterol prevented the phorbol ester-stimulated localization of Rac1 to the plasma membrane. Thus, our results demonstrate that cholesterol is required for the membrane localization of activated Rac1, actin reorganization, membrane ruffling and macropinocytosis. PMID- 12082156 TI - The minor regulated pathway, a rapid component of salivary secretion, may provide docking/fusion sites for granule exocytosis at the apical surface of acinar cells. AB - Recently, we reported that the minor regulated and constitutive-like pathways are the main source of resting secretion by parotid acinar cells. Using tissue lobules biosynthetically labeled with [(35)S]amino acids, we now show that discharge of the minor regulated pathway precedes granule exocytosis stimulated by isoproterenol (> or =1 microM) or carbachol (2 microM). Stimulation of the minor regulated pathway by 40 nM carbachol as well as altering its trafficking, either by adding brefeldin A or by incubating in K(+)-free medium, cause potentiation of amylase secretion stimulated by isoproterenol, suggesting that the minor regulated pathway contributes to the mechanism of potentiation. Both exocytosis of the minor regulated pathway and the potentiation-inducing treatments induce relocation of immunostained subapical puncta of the SNARE protein syntaxin 3 into the apical plasma membrane. Rab11 and possibly VAMP2 may be concentrated in the same relocating foci. These results suggest that the minor regulated pathway and granule exocytosis are functionally linked and that the minor regulated pathway has a second role beyond contributing to resting secretion - providing surface docking/fusion sites for granule exocytosis. In the current model of salivary protein export, discharge of the minor regulated pathway by either beta-adrenergic or cholinergic stimulation is an obligatory first step. Ensuing granule exocytosis is controlled mainly by beta-adrenergic stimulation whereas cholinergic stimulation mainly regulates the number of surface sites where release occurs. PMID- 12082157 TI - Granule-specific ATP requirements for Ca2+-induced exocytosis in human neutrophils. Evidence for substantial ATP-independent release. AB - Ca2+-induced exocytosis in neuronal and neuroendocrine cells involves ATP dependent steps believed to 'prime' vesicles for exocytosis. Primed, docked vesicles are released in response to Ca2+ influx through voltage-gated Ca2+ channels. Neutrophils, however, do not possess voltage-gated Ca2+ channels and appear to have no docked vesicles. Furthermore, neutrophils have several types of granules with markedly different Ca2+ requirements for exocytosis. These differential Ca2+ dependencies were used as a tool to investigate the ATP dependence of different granule populations. Here we demonstrate distinct ATP requirements for release of neutrophil granule populations, with respect to rate as well as amplitude. Intracellular ATP was depleted to various levels, and exocytosis was stimulated with different Ca2+ concentrations and measured with the patch-clamp capacitance technique or by detecting enzyme release. Primary granule exocytosis displayed a distinct ATP dependence with an apparent K(m) of approximately 80 microM ATP and no ATP-independent exocytosis. Release of secondary and tertiary granules displayed a more shallow ATP dependence (K(m) approximately 330 microM), and more than 50% of secondary and tertiary granules appeared not to need ATP at all for their release. Individual granules in human neutrophils have distinct ATP requirements for exocytosis, suggesting that the ATP-sensitive elements are localised to the granules. Primary granule exocytosis has a very low affinity for ATP. Furthermore, substantial ATP-independent exocytosis of secondary and tertiary granule occurs despite the absence of docked granules. These characteristics should help neutrophils to fulfil their bactericidal functions at poorly irrigated sites of infection with low glucose supply. PMID- 12082158 TI - Requirements for the nuclear export of the small ribosomal subunit. AB - Eukaryotic ribosome biogenesis requires multiple steps of nuclear transport because ribosomes are assembled in the nucleus while protein synthesis occurs in the cytoplasm. Using an in situ RNA localization assay in the yeast Saccharomyces cerevisiae, we determined that efficient nuclear export of the small ribosomal subunit requires Yrb2, a factor involved in Crm1-mediated export. Furthermore, in cells lacking YRB2, the stability and abundance of the small ribosomal subunit is decreased in comparison with the large ribosomal subunit. To identify additional factors affecting small subunit export, we performed a large-scale screen of temperature-sensitive mutants. We isolated new alleles of several nucleoporins and Ran-GTPase regulators. Together with further analysis of existing mutants, we show that nucleoporins previously shown to be defective in ribosomal assembly are also defective in export of the small ribosomal subunit. PMID- 12082159 TI - The molecular mechanism of translocation through the nuclear pore complex is highly conserved. AB - In this report we investigated the activity of vertebrate nuclear transport factors in a primitive organism, Amoeba proteus, to better understand evolutionary changes in the transport mechanisms of organisms expected to have different requirements for nucleocytoplasmic exchange. It was initially determined that FxFG-containing nucleoporins and Ran, both of which are essential for nuclear import in vertebrates, as well as yeast, are also present and functional in amoebae. This suggests that there are fundamental similarities in the transport process; however, there are also significant differences. Transport substrates containing either the hnRNP A1 M9 shuttling signal (a GST/GFP/M9 fusion protein) or the classical bipartite NLS (colloidal gold coated with BSA bipartite NLS conjugates), both of which are effectively transported in vertebrate cells, are excluded from the nucleus when microinjected into amoebae. However, when these substrates are injected along with transportin or importin alpha/beta, respectively, the vertebrate receptors for these signals, they readily accumulate in the nucleoplasm. These results indicate that although the molecular recognition of substrates is not well conserved between vertebrates and amoebae, vertebrate transport receptors are functional in A. proteus, showing that the translocation machinery is highly conserved. Since selected nuclear import pathways can be investigated in the absence of competing endogenous transport, A. proteus might provide a useful in vivo system for investigating specific molecular interactions involved in trafficking. PMID- 12082160 TI - A role for mammalian Ubc6 homologues in ER-associated protein degradation. AB - Integral membrane and secretory proteins which fail to fold productively are retained in the endoplasmic reticulum and targeted for degradation by cytoplasmic proteasomes. Genetic and biochemical analyses suggest that substrates of this pathway must be dislocated across the membrane of the endoplasmic reticulum (ER) by a process requiring a functional Sec61 complex and multiubiquitinylation. In yeast, the tail-anchored ubiquitin-conjugating enzyme Ubc6p, which is localized to the cytoplasmic surface of the ER, participates in ER-associated degradation (ERAD) of misfolded proteins. Here we describe the identification of two families of mammalian Ubc6p-related proteins. Members of both families are also located in the ER membrane and display a similar membrane topology as the yeast enzyme. Furthermore we show that expression of elevated levels of wild-type and dominant negative alleles of these components affects specifically ERAD of the alpha subunit of the T-cell receptor and a mutant form of the CFTR protein. Similarly, we describe that the expression level of Ubc6p in yeast is also critical for ERAD, suggesting that the Ubc6p function is highly conserved from yeast to mammals. PMID- 12082162 TI - Predicting the transmembrane secondary structure of ligand-gated ion channels. AB - Recent mutational analyses of ligand-gated ion channels (LGICs) have demonstrated a plausible site of anesthetic action within their transmembrane domains. Although there is a consensus that the transmembrane domain is formed from four membrane-spanning segments, the secondary structure of these segments is not known. We utilized 10 state-of-the-art bioinformatics techniques to predict the transmembrane topology of the tetrameric regions within six members of the LGIC family that are relevant to anesthetic action. They are the human forms of the GABA alpha 1 receptor, the glycine alpha 1 receptor, the 5HT3 serotonin receptor, the nicotinic AChR alpha 4 and alpha 7 receptors and the Torpedo nAChR alpha 1 receptor. The algorithms utilized were HMMTOP, TMHMM, TMPred, PHDhtm, DAS, TMFinder, SOSUI, TMAP, MEMSAT and TOPPred2. The resulting predictions were superimposed on to a multiple sequence alignment of the six amino acid sequences created using the CLUSTAL W algorithm. There was a clear statistical consensus for the presence of four alpha helices in those regions experimentally thought to span the membrane. The consensus of 10 topology prediction techniques supports the hypothesis that the transmembrane subunits of the LGICs are tetrameric bundles of alpha helices. PMID- 12082161 TI - Ihh enhances differentiation of CFK-2 chondrocytic cells and antagonizes PTHrP mediated activation of PKA. AB - Indian Hedgehog (Ihh), a member of the hedgehog (HH) family of secreted morphogens, and parathyroid hormone-related peptide (PTHrP) are key regulators of cartilage cell (chondrocyte) differentiation. We have investigated, in vitro, the actions of HH signalling and its possible interplay with PTHrP using rat CFK-2 chondrocytic cells. Markers of chondrocyte differentiation [alkaline phosphatase (ALP) activity, and type II (Col2a1) and type X collagen (Col10a1) expression] were enhanced by overexpression of Ihh or its N-terminal domain (N-Ihh), effects mimicked by exogenous administration of recombinant N-terminal HH peptide. Moreover, a missense mutation mapping to the N-terminal domain of Ihh (W160G) reduces the capacity of N-Ihh to induce differentiation. Prolonged exposure of CFK-2 cells to exogenous N-Shh (5x10(-9) M) in the presence of PTHrP (10(-8) M) or forskolin (10(-7) M) resulted in perturbation of HH-mediated differentiation. In addition, overexpression of a constitutively active form of the PTHrP receptor (PTHR1 H223R) inhibited Ihh-mediated differentiation, implicating activation of protein kinase A (PKA) by PTHR1 as a probable mediator of the antagonistic effects of PTHrP. Conversely, overexpression of Ihh/N-Ihh or exogenous treatment with N-Shh led to dampening of PTHrP-mediated activation of PKA. Taken together, our data suggest that Ihh harbors the capacity to induce rather than inhibit chondrogenic differentiation, that PTHrP antagonizes HH-mediated differentiation through a PKA-dependent mechanism and that HH signalling, in turn, modulates PTHrP action through functional inhibition of signalling by PTHR1 to PKA. PMID- 12082163 TI - Thermodynamic characterization of variants of mesophilic cytochrome c and its thermophilic counterpart. AB - Thermal stability was measured for variants of cytochrome c-551 (PA c-551) from a mesophile, Pseudomonas aeruginosa, and a thermophilic counterpart, Hydrogenobacter thermophilus cytochrome c-552 (HT c-552), by differential scanning calorimetry (DSC) at pH 3.6. The mutated residues in PA c-551, selected with reference to the corresponding residues in HT c-552, were located in three spatially separated regions: region I, Phe7 to Ala/Val13 to Met; region II, Glu34 to Tyr/Phe43 to Tyr; and region III, Val78 to Ile. The thermodynamic parameters determined indicated that the mutations in regions I and III caused enhanced stability through not only enthalpic but also entropic contributions, which reflected improved packing of the side chains. Meanwhile, the mutated region II made enthalpic contributions to the stability through electrostatic interactions. The obtained differences in the Gibbs free energy changes of unfolding [Delta(DeltaG)] showed that the three regions contributed to the overall stability in an additive manner. HT c-552 had the smallest heat capacity change (DeltaC(P)), resulting in higher DeltaG values over a wide temperature range (0 100 degrees C), compared to the PA c-551 variants; this contributed to the highest stability of HT c-552. Our DSC measurement results, in conjunction with mutagenesis and structural studies on the homologous mesophilic and thermophilic cytochromes c, provided an extended thermodynamic view of protein stabilization. PMID- 12082164 TI - Improving a circularly permuted TEM-1 beta-lactamase by directed evolution. AB - Circular permutation of proteins is a powerful technique to explore the importance of the polypeptide secondary structure order for attaining the final three-dimensional structure. Here, we designed a circular permutation of the TEM beta-lactamase in order to produce a new domain-forming amino acid arrangement in the polypeptide sequence. Closing the normal N- and C-termini with the connecting peptide GGS and creating new N- and C-termini at position 216, produces a severely impaired permuted protein. Introduction of a connector with random components allows the isolation of enzymes with better activities and indicates a selection for a potential helix-stop signal at the new super-secondary motif. We applied several directed-evolution cycles, starting from permuted enzymes with each of the two different connecting peptides, and selecting for antibiotic resistance and isolated several mutants with resistance levels close to those of the wild-type enzyme. We also analyze some of the data collected on the outcomes and paths of these evolutionary experiments. A purified sixth cycle variant with connector peptide GGS showed catalytic efficiency values approximately 8% of the natural enzyme. PMID- 12082165 TI - Cold-adaptation mechanism of mutant enzymes of 3-isopropylmalate dehydrogenase from Thermus thermophilus. AB - Random mutagenesis of Thermus thermophilus 3-isopropylmalate dehydrogenase revealed that a substitution of Val126Met in a hinge region caused a marked increase in specific activity, particularly at low temperatures, although the site is far from the binding residues for 3-isopropylmalate and NAD. To understand the molecular mechanism, residue 126 was substituted with one of eight other residues, Gly, Ala, Ser, Thr, Glu, Leu, Ile or Phe. Circular dichroism analyses revealed a decreased thermal stability of the mutants (Delta T ((1/2))= 0-13 degrees C), indicating structural perturbations caused by steric conflict with surrounding residues having larger side chains. Kinetic parameters, k(cat) and K(m) values for isopropylmalate and NAD, were also affected by the mutation, but the resulting k(cat)/K(m) values were similar to that of the wild-type enzyme, suggesting that the change in the catalytic property is caused by the change in free-energy level of the Michaelis complex state relative to that of the initial state. The kinetic parameters and activation enthalpy change (Delta H (double dagger)) showed good correlation with the van der Waals volume of residue 126. These results suggested that the artificial cold adaptation (enhancement of k(cat) value at low temperatures) resulted from the destabilization of the ternary complex caused by the increase in the volume of the residue at position 126. PMID- 12082166 TI - Alteration of substrate specificity of cholesterol oxidase from Streptomyces sp. by site-directed mutagenesis. AB - Despite the structural similarities between cholesterol oxidase from Streptomyces and that from Brevibacterium, both enzymes exhibit different characteristics, such as catalytic activity, optimum pH and temperature. In attempts to define the molecular basis of differences in catalytic activity or stability, substitutions at six amino acid residues were introduced into cholesterol oxidase using site directed mutagenesis of its gene. The amino acid substitutions chosen were based on structural comparisons of cholesterol oxidases from Streptomyces and BREVIBACTERIUM: Seven mutant enzymes were constructed with the following amino acid substitutions: L117P, L119A, L119F, V145Q, Q286R, P357N and S379T. All the mutant enzymes exhibited activity with the exception of that with the L117P mutation. The resulting V145Q mutant enzyme has low activities for all substrates examined and the S379T mutant enzyme showed markedly altered substrate specificity compared with the wild-type enzyme. To evaluate the role of V145 and S379 residues in the reaction, mutants with two additional substitutions in V145 and four in S379 were constructed. The mutant enzymes created by the replacement of V145 by Asp and Glu had much lower catalytic efficiency for cholesterol and pregnenolone as substrates than the wild-type enzyme. From previous studies and this study, the V145 residue seems to be important for the stability and substrate binding of the cholesterol oxidase. In contrast, the catalytic efficiencies (k(cat)/K(m)) of the S379T mutant enzyme for cholesterol and pregnenolone were 1.8- and 6.0-fold higher, respectively, than those of the wild type enzyme. The enhanced catalytic efficiency of the S379T mutant enzyme for pregnenolone was due to a slightly high k(cat) value and a low K(m) value. These findings will provide several ideas for the design of more powerful enzymes that can be applied to clinical determination of serum cholesterol levels and as sterol probes. PMID- 12082167 TI - Rational design of 'water-soluble' bacteriorhodopsin variants. AB - We have explored the interchangeability of soluble and membrane proteins by attempting to render a helical membrane protein 'water soluble' through mutation of its lipid-exposed residues. Using an atomic resolution structure of bacteriorhodopsin (bR), two different strategies were developed to identify lipid exposed residues for mutation. In the first strategy all residues in trimeric bR with solvent accessibility >35% were marked for replacement. Replacement residues were chosen so as to map an average surface of helical soluble proteins onto the bR surface, resulting in the mutagenesis of 14.9% of surface residues. The second strategy took into account the observation that accessible residues can be categorized as fully or partially accessible. Consequently, three mutants were designed based on monomeric bR, all with their accessible residues changed and with varying extents of mutagenesis of partially accessible residues. 13.5-24.3% of the wild-type surface was altered in these designs. The construct for the first design was cloned into Escherichia coli. Trace amounts of the mutant protein were expressed with the concurrent overexpression of an endogenous prolyl isomerase. In contrast, all three mutant proteins of the second design expressed well and could be purified to homogeneity. Systematic refolding trials were undertaken with limited success at solubilization in aqueous media. We have discussed the feasibility of applying the 'solubilization strategy' outlined here to membrane proteins. PMID- 12082168 TI - Rational design of green fluorescent protein mutants as biosensor for bacterial endotoxin. AB - Enhanced green fluorescent protein (EGFP) was selected as a signalling scaffold protein for design of a fluorescent biosensor for bacterial endotoxin [or lipopolysaccharide (LPS)]. Virtual mutagenesis was utilized to model EGFP variants containing binding sites for LPS and lipid A (LA), the bioactive component of LPS. Cationic amphipathic sequences of five alternating basic and hydrophobic residues were introduced to beta-sheets located on the surface of EGFP barrel, in the vicinity of the chromophore. Computational methods were employed to predict binding affinity of Escherichia coli LA, to the models of virtual EGFP mutants. DNA mutant constructs of five predicted best binding EGFP variants were expressed in COS-1 cells. The EGFP-mutant proteins exhibited differential expression and variable degrees of fluorescence yield at 508 nm. The EGFP mutants showed a range of LA binding affinities that corresponded to the computational predictions. LPS/LA binding to the mutants caused concentration dependent fluorescence quenching. The EGFP mutant, G10 bearing LPS/LA amphipathic binding motif in the vicinity of the chromophore (YLSTQ(200-204)-->KLKTK) captured LA with a dissociation constant of 8.5 microm. G10 yielded the highest attenuation of fluorescence intensity in the presence of LPS/LA and demonstrated capability in fluorescence-mediated quantitative detection of LPS in endotoxin contaminated samples. Thus, the EGFP mutant can form the basis of a novel fluorescent biosensor for bacterial endotoxin. PMID- 12082169 TI - Proteomics analysis of carbon-starved Mycobacterium smegmatis: induction of Dps like protein. AB - Mycobacterium tuberculosis is a globally successful pathogen, infecting more than one third of total world's population. These bacteria have the remarkable ability to persist in the host for long periods of time unrecognized by the immune system and then to re-emerge later in life causing the disease. The physiology of such persistent or dormant bacilli is not very well characterized. Some evidence suggests that the dormant bacilli survive in a nutrient-deprived state that is similar to the stationary phase of the bacteria with respect to gene expression and physiology. Under this assumption we have studied the survival of Mycobacterium smegmatis in carbon starvation conditions as a model for mycobacterial persistence. M.smegmatis, being a fast-growing strain, serves as a good model to study starvation responses. Using the two-dimensional electrophoresis-based proteomics approach, we identified a protein which was found to be expressed preferentially under starvation conditions. This protein is homologous to a family of proteins called Dps (DNA binding Protein from Starved cells) that are known to protect DNA under various kinds of environmental stresses and its existence has, so far, not been reported in mycobacteria. Upon expression and purification of this protein, we observed that it has non-specific DNA-binding ability. Formation of a cage-like dodecamer structure is a characteristic feature of Dps. Using comparative modelling we were able to show that Dps from M.smegmatis could form a dodecamer structure similar to the crystal structure of Dps from Escherichia coli. PMID- 12082170 TI - Lipid-interacting properties of the N-terminal domain of human apolipoprotein C III. AB - The lipid-interacting properties of the N-terminal domain of human apolipoprotein C-III (apo C-III) were investigated. By molecular modeling, we predicted that the 6-20 fragment of apo C-III is obliquely orientated at the lipid/water interface owing to an asymmetric distribution of the hydrophobic residues when helical. This is characteristic of 'tilted peptides' originally discovered in viral fusion proteins and later in various proteins including some involved in lipoprotein metabolism. Since most tilted peptides were shown to induce liposome fusion in vitro, the fusogenic capacity of the 6-20 fragment of apo C-III was tested on unilamellar liposomes and compared with the well characterized SIV fusion peptide. Mutants were designed by molecular modeling to assess the role of the hydrophobicity gradient in the fusion. FTIR spectroscopy confirmed the predominantly helical conformation of the peptides in TFE solution and also in lipid-peptide complexes. Lipid-mixing experiments showed that the apo C-III (6 20) peptide is able to increase the fluorescence of a lipophilic fluorescent probe. The vesicle fusion was confirmed by core-mixing and leakage assays. The hydrophobicity gradient plays a key role in the fusion process because the mutant with no hydrophobic asymmetry but the same mean hydrophobicity as the wild type does not induce significant lipid fusion. The apo C-III (6-20) fragment is, however, less fusogenic than the SIV peptide, in agreement with their respective mean hydrophobicity. Since lipid fusion should not be the physiological function of the N-terminal domain of apo CIII, we suggest that its peculiar distribution of hydrophobic residues is important for the lipid-binding properties of apo C III and should be involved in apolipoprotein and lipid exchanges crucial for triglyceride metabolism. PMID- 12082171 TI - Role of paired basic residues of protein C-termini in phospholipid binding. AB - It is a well known phenomenon that the occurrence of several distinct amino acids at the C-terminus of proteins is non-random. We have analysed all Saccharomyces cerevisiae proteins predicted by computer databases and found lysine to be the most frequent residue both at the last (-1) and at the penultimate amino acid ( 2) positions. To test the hypothesis that C-terminal basic residues efficiently bind to phospholipids we randomly expressed GST-fusion proteins from a yeast genomic library. Fifty-four different peptide fragments were found to bind phospholipids and 40% of them contained lysine/arginine residues at the (-1) or ( 2) positions. One peptide showed high sequence similarity with the yeast protein Sip18p. Mutational analysis revealed that both C-terminal lysine residues of Sip18p are essential for phospholipid-binding in vitro. We assume that basic amino acid residues at the (-1) and (-2) positions in C-termini are suitable to attach the C-terminus of a given protein to membrane components such as phospholipids, thereby stabilizing the spatial structure of the protein or contributing to its subcellular localization. This mechanism could be an additional explanation for the C-terminal amino acid bias observed in proteins of several species. PMID- 12082172 TI - High-level expression and characterization of a glycosylated covalently linked dimer of the prion protein. AB - There is evidence that prion protein dimers may be involved in the formation of the scrapie prion protein, PrP(Sc), from its normal (cellular) form, PrP(c). Recently, the crystal structure of the human prion protein in a dimeric form was reported. Here we report for the first time the overexpression of a human PrP dimer covalently linked by a FLAG peptide (PrP::FLAG::PrP) in the methylotrophic yeast Pichia pastoris. FLAG-tagged human PrP (aa1-aa253) (huPrP::FLAG) was also expressed in the same system. Treatment with tunicamycin and endoglycosidase H showed that both fusion proteins are expressed as various glycoforms. Both PrP proteins were completely digested by proteinase K (PK), suggesting that the proteins do not have a PrP(Sc) structure and are not infectious. Plasma membrane fractionation revealed that both proteins are transported to the plasma membrane of the cell. The glycosylated proteins might act as powerful tools for crystallization trials, PrP(c)/PrP(Sc) conversion studies and other applications in the life cycle of prions. PMID- 12082173 TI - Waddington's canalization revisited: developmental stability and evolution. AB - Most species maintain abundant genetic variation and experience a range of environmental conditions, yet phenotypic variation is low. That is, development is robust to changes in genotype and environment. It has been claimed that this robustness, termed canalization, evolves because of long-term natural selection for optimal phenotypes. We show that the developmental process, here modeled as a network of interacting transcriptional regulators, constrains the genetic system to produce canalization, even without selection toward an optimum. The extent of canalization, measured as the insensitivity to mutation of a network's equilibrium state, depends on the complexity of the network, such that more highly connected networks evolve to be more canalized. We argue that canalization may be an inevitable consequence of complex developmental-genetic processes and thus requires no explanation in terms of evolution to suppress phenotypic variation. PMID- 12082174 TI - PRKX, a phylogenetically and functionally distinct cAMP-dependent protein kinase, activates renal epithelial cell migration and morphogenesis. AB - The human protein kinase X gene (PRKX) is a member of an ancient family of cAMP dependent serine/threonine kinases here shown to be phylogenetically distinct from the classical PKA, PKB/Akt, PKC, SGK, and PKG gene families. Renal expression of the PRKX gene is developmentally regulated and restricted to the ureteric bud epithelium of the fetal metanephric kidney. Aberrant adult kidney expression of PRKX was found in autosomal dominant polycystic kidney disease. PRKX kinase expression markedly activated migration of cultured renal epithelial cells in the presence of cAMP; this effect was blocked by cell treatment with the PKA inhibitor H89 and was not observed in PKA-transfected cells. In addition, expression of PRKX kinase activated branching morphogenesis of Madin-Darby canine kidney cells in collagen gels even in the absence of cAMP and/or hepatocyte growth factor, an effect not seen with either PKA expression or expression of a mutant, kinase-inactivated PRKX. These results suggest that the PRKX kinase may regulate epithelial morphogenesis during mammalian kidney development. Because another member of the PRKX gene family (the Dictyostelium discoideum gene KAPC DICDI) also plays a role in cellular migration, these studies suggest that regulation of morphogenesis may be a distinctive property of these genes that has been conserved in evolution that is not shared with PKA family genes. PMID- 12082175 TI - An estrogen receptor repressor induces cataract formation in transgenic mice. AB - Despite the high prevalence of age-related cataracts, there are currently no known therapies to delay or prevent their occurrence. Studies in humans and rodent models suggest that estrogen may provide protection against age-related cataracts. The discovery of ocular estrogen receptors (ERs) indicates that estrogen protection may result from direct interactions with its receptors in the eye, instead an indirect consequence from effects on another tissue. Studies in our transgenic mouse model validate the concept that estrogen is beneficial for the eye. These mice express ER Delta , a dominant-negative form of ER alpha that inhibits ER alpha function. In the ER Delta 3 transgenic mice, cortical cataracts spontaneously form in ER Delta 3 females after puberty and progress with age. The cataracts initiate in the equatorial region of the lens where the epithelial cells differentiate into elongating fiber cells. Cataract formation can be prevented if the females are ovariectomized before, but not after, sexual maturity. Both male and female ER Delta 3 mice develop cataracts after neonatal treatment with the potent estrogen diethylstilbestrol (DES). The incidence of spontaneous and DES-induced cataracts in ER Delta 3 mice is 100%, yet these cataracts are absent from the wild-type mice. These data suggest that repression of estrogen action induces cortical cataract formation because estrogen is required to activate the ER Delta 3 repressor. Evidence of DES-induced cataracts in the ER Delta 3 males as well as the females suggests that estrogen is important in lens physiology in both sexes. The ER Delta 3 mice provide a powerful model for assessing the role of estrogen in maintaining the transparency of the lens. PMID- 12082177 TI - Diffuse fluid flux through orogenic belts: implications for the world ocean. AB - Fifty years ago a classic paper by W. W. Rubey [(1951) Geol. Soc. Am. Bull. 62, 1111-1148] examined various hypotheses regarding the origin of sea water and concluded that the most likely hypothesis was volcanic outgassing, a view that was generally accepted by earth scientists for the next several decades. More recent work suggests that the rate of subduction of water is much larger than the volcanic outgassing rate, lending support to hypotheses that either ocean volume has decreased with time, or that the imbalance is offset by continuous replenishment of water by cometary impacts. These alternatives are required in the absence of additional mechanisms for the return of water from subducting lithosphere to the Earth's surface. Our recent work on crustal permeability suggests a large capacity for water upflow through tectonically active continental crust, resulting in a heretofore unrecognized degassing pathway that can accommodate the water subduction rate. Escape of recycled water via delivery from the mantle through zones of active metamorphism eliminates the mass-balance argument for the loss of ocean volume or extraterrestrial sources. PMID- 12082176 TI - Three-dimensional structure of the amino-terminal domain of syntaxin 6, a SNAP-25 C homolog. AB - Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins are required for intracellular membrane fusion, and are differentially localized throughout the cell. SNAREs on vesicle and target membranes contain "SNARE motifs" which interact to form a four-helix bundle that contributes to the fusion of two membranes. SNARE motif sequences fall into four classes, homologous to the neuronal proteins syntaxin 1a, VAMP 2, and the N- and C-terminal SNARE motifs of SNAP-25 (S25N and S25C), and it is thought that one member from each class interacts to form a SNARE complex. Many SNAREs also feature N-terminal domains believed to function in regulating SNARE complex assembly or other aspects of vesicle transport. Syntaxin 6 is a SNARE found primarily in endosomal transport vesicles and whose SNARE motif shows significant homology to both syntaxin 1a and S25C. The crystal structure of the syntaxin 6 N-terminal domain reveals strong structural similarity with the N-terminal domains of syntaxin family members syntaxin 1a, Sso1p, and Vam3p, despite a very low level of sequence similarity. The syntaxin 6 SNARE motif can substitute for S25C in in vitro binding experiments, supporting the classification of syntaxin 6 as an S25C family member. Secondary structure prediction of SNARE proteins shows that the N terminal domains of many syntaxin, S25N, and S25C family members are likely to be similar to one another, but are distinct from those of VAMP family members, indicating that syntaxin, S25N, and S25C SNAREs may have shared a common ancestor. PMID- 12082178 TI - Apparent role of traveling metabolic waves in oxidant release by living neutrophils. AB - Cell metabolism self-organizes into two types of dissipative structures: chemical oscillations and traveling metabolic waves. In the present study we test the hypothesis that traveling NAD(P)H waves within neutrophils are associated spatially and temporally with the release of reactive oxygen metabolites (ROMs). Using high-speed optical microscopy and taking advantage of the autofluorescence of NAD(P)H, we have observed the propagation of NAD(P)H waves within cells. When NAD(P)H waves reach the lamellipodium of morphologically polarized neutrophils, a diffusing plume of superoxide is released as evidenced by the conversion of hydroethidine in the extracellular environment to ethidium bromide. Parallel results were obtained by using high-speed emission microspectrophotometry. These experiments indicate that the spatial and temporal properties of NAD(P)H waves are transformed into ROM pulses in the extracellular environment. Propagating NAD(P)H waves allow neutrophils to specifically deliver substrate to the lamellipodium at high concentrations, thus facilitating the local and periodic release of ROMs in the direction of cell movement and/or a target. PMID- 12082179 TI - Cluster analysis of gene expression dynamics. AB - This article presents a Bayesian method for model-based clustering of gene expression dynamics. The method represents gene-expression dynamics as autoregressive equations and uses an agglomerative procedure to search for the most probable set of clusters given the available data. The main contributions of this approach are the ability to take into account the dynamic nature of gene expression time series during clustering and a principled way to identify the number of distinct clusters. As the number of possible clustering models grows exponentially with the number of observed time series, we have devised a distance based heuristic search procedure able to render the search process feasible. In this way, the method retains the important visualization capability of traditional distance-based clustering and acquires an independent, principled measure to decide when two series are different enough to belong to different clusters. The reliance of this method on an explicit statistical representation of gene expression dynamics makes it possible to use standard statistical techniques to assess the goodness of fit of the resulting model and validate the underlying assumptions. A set of gene-expression time series, collected to study the response of human fibroblasts to serum, is used to identify the properties of the method. PMID- 12082180 TI - Conformational change of the actomyosin complex drives the multiple stepping movement. AB - Actin-myosin (actomyosin) generates mechanical force by consuming ATP molecules. We apply the energy landscape perspective to address a controversial issue as to whether the myosin head moves with multiple steps after a single ATP hydrolysis or only a single mechanical event of the lever-arm swinging follows a single ATP hydrolysis. Here we propose a theoretical model in which the refolding of the partially unfolded actomyosin complex and the movement of the myosin head along the actin filament are coupled. A single ATP hydrolysis is followed by the formation of a high free-energy partially unfolded actomyosin complex, which then gradually refolds with a concomitant multiple stepping movement on the way to the lowest free-energy rigor state. The model quantitatively explains the single molecular observation of the multiple stepping movement and is consistent with structural observations of the disorder in the actomyosin-binding process. The model also explains the observed variety in dwell time before each step, which is not accounted for by previous models, such as the lever-arm or ratchet models. PMID- 12082181 TI - Tissue-nonspecific alkaline phosphatase and plasma cell membrane glycoprotein-1 are central antagonistic regulators of bone mineralization. AB - Osteoblasts mineralize bone matrix by promoting hydroxyapatite crystal formation and growth in the interior of membrane-limited matrix vesicles (MVs) and by propagating the crystals onto the collagenous extracellular matrix. Two osteoblast proteins, tissue-nonspecific alkaline phosphatase (TNAP) and plasma cell membrane glycoprotein-1 (PC-1) are involved in this process. Mutations in the TNAP gene result in the inborn error of metabolism known as hypophosphatasia, characterized by poorly mineralized bones, spontaneous fractures, and elevated extracellular concentrations of inorganic pyrophosphate (PP(i)). PP(i) suppresses the formation and growth of hydroxyapatite crystals. PP(i) is produced by the nucleoside triphosphate pyrophosphohydrolase activity of a family of isozymes, with PC-1 being the only member present in MVs. Mice with spontaneous mutations in the PC-1 gene have hypermineralization abnormalities that include osteoarthritis and ossification of the posterior longitudinal ligament of the spine. Here, we show the respective correction of bone mineralization abnormalities in knockout mice null for both the TNAP (Akp2) and PC-1 (Enpp1) genes. Each allele of Akp2 and Enpp1 has a measurable influence on mineralization status in vivo. Ex vivo experiments using cultured double-knockout osteoblasts and their MVs demonstrate normalization of PP(i) content and mineral deposition. Our data provide evidence that TNAP and PC-1 are key regulators of the extracellular PP(i) concentrations required for controlled bone mineralization. Our results suggest that inhibiting PC-1 function may be a viable therapeutic strategy for hypophosphatasia. Conversely, interfering with TNAP activity may correct pathological hyperossification because of PP(i) insufficiency. PMID- 12082182 TI - Randomized controlled trial of a treatment for anorexia and bulimia nervosa. AB - Evidence for the effectiveness of existing treatments of patients with eating disorders is weak. Here we describe and evaluate a method of treatment in a randomized controlled trial. Sixteen patients, randomly selected out of a group composed of 19 patients with anorexia nervosa and 13 with bulimia nervosa, were trained to eat and recognize satiety by using computer support. They rested in a warm room after eating, and their physical activity was restricted. The patients in the control group (n = 16) received no treatment. Remission was defined by normal body weight (anorexia), cessation of binge eating and purging (bulimia), a normal psychiatric profile, normal laboratory test values, normal eating behavior, and resumption of social activities. Fourteen patients went into remission after a median of 14.4 months (range 4.9-26.5) of treatment, but only one patient went into remission while waiting for treatment (P = 0.0057). Relapse is considered a major problem in patients who have been treated to remission. We therefore report results on a total of 168 patients who have entered our treatment program. The estimated rate of remission was 75%, and estimated time to remission was 14.7 months (quartile range 9.6 > or = 32). Six patients (7%) of 83 who were treated to remission relapsed, but the others (93%) have remained in remission for 12 months (quartile range 6-36). Because the risk of relapse is maximal in the first year after remission, we suggest that most patients treated with this method recover. PMID- 12082183 TI - Restricted islet-cell reactive T cell repertoire of early pancreatic islet infiltrates in NOD mice. AB - The mechanisms responsible for initiating autoimmune diabetes remain obscure. Here, we describe a method for identifying both the alpha- and beta-chains of the T cell receptor (TCR) from individual pancreatic islet-infiltrating T cells at the earliest stages of disease in nonobese diabetic mice (NOD). Analysis of the TCR repertoire of these early islet infiltrates reveals enrichment for a small subset of TCR sequences. Reconstitution of these TCR in vitro demonstrates that these receptors confer reactivity to islet cells but not to the well characterized autoantigens, glutamic acid decarboxylase (GAD65) and insulin. Thus, autoimmune diabetes in NOD may be initiated by a limited number of antigens distinct from GAD65 and insulin. PMID- 12082184 TI - Recombinant human erythropoietin counteracts secondary injury and markedly enhances neurological recovery from experimental spinal cord trauma. AB - Erythropoietin (EPO) functions as a tissue-protective cytokine in addition to its crucial hormonal role in red cell production. In the brain, for example, EPO and its receptor are locally produced, are modulated by metabolic stressors, and provide neuroprotective and antiinflammatory functions. We have previously shown that recombinant human EPO (rhEPO) administered within the systemic circulation enters the brain and is neuroprotective. At present, it is unknown whether rhEPO can also improve recovery after traumatic injury of the spinal cord. To evaluate whether rhEPO improves functional outcome if administered after cord injury, two rodent models were evaluated. First, a moderate compression of 0.6 N was produced by application of an aneurysm clip at level T3 for 1 min. RhEPO (1,000 units per kg of body weight i.p.) administered immediately after release of compression was associated with partial recovery of motor function within 12 h after injury, which was nearly complete by 28 days. In contrast, saline-treated animals exhibited only poor recovery. In the second model used, rhEPO administration (5,000 units per kg of body weight i.p. given once 1 h after injury) also produced a superior recovery of function compared with saline-treated controls after a contusion of 1 N at level T9. In this model of more severe spinal cord injury, secondary inflammation was also markedly attenuated by rhEPO administration and associated with reduced cavitation within the cord. These observations suggest that rhEPO provides early recovery of function, especially after spinal cord compression, as well as longer-latency neuroprotective, antiinflammatory and antiapoptotic functions. PMID- 12082185 TI - The aging baboon: comparative demography in a non-human primate. AB - Why do closely related primate genera vary in longevity, and what does this teach us about human aging? Life tables of female baboons (Papio hamadryas) in two wild populations of East Africa and in a large captive population in San Antonio, Texas, provide striking similarities and contrasts to human mortality patterns. For captive baboons at the Southwest Foundation for Biomedical Research, we estimate the doubling time of adult mortality rate as 4.8 years. Wild females in free-living populations in Tanzania and in Kenya showed doubling times of 3.5 and 3.8 years, respectively. Although these values are considerably faster than the estimates of 7-8 years for humans, these primates share a demographic feature of human aging: within each taxon populations primarily vary in the level of Gompertz mortality intercept (frailty) and vary little in the demographic rate of aging. Environmental and genetic factors within taxa appear to affect the level of frailty underlying senescence. In contrast, primate taxa are differentiated by rates of demographic aging, even if they cannot be characterized by species specific lifespan. PMID- 12082186 TI - Eye contact detection in humans from birth. AB - Making eye contact is the most powerful mode of establishing a communicative link between humans. During their first year of life, infants learn rapidly that the looking behaviors of others conveys significant information. Two experiments were carried out to demonstrate special sensitivity to direct eye contact from birth. The first experiment tested the ability of 2- to 5-day-old newborns to discriminate between direct and averted gaze. In the second experiment, we measured 4-month-old infants' brain electric activity to assess neural processing of faces when accompanied by direct (as opposed to averted) eye gaze. The results show that, from birth, human infants prefer to look at faces that engage them in mutual gaze and that, from an early age, healthy babies show enhanced neural processing of direct gaze. The exceptionally early sensitivity to mutual gaze demonstrated in these studies is arguably the major foundation for the later development of social skills. PMID- 12082187 TI - Current clinical characteristics and economic impact of subacute stent thrombosis. AB - Subacute stent thrombosis (SAT), while uncommon, continues to produce serious clinical consequences, including major myocardial infarction (MI) in 60 70% of cases, and short-term mortality rates of 20% or higher. A number of procedural and patient factors have been shown to predict the occurrence of SAT, including longer stent length, smaller minimum luminal diameter, persistent dissection, multivessel intervention, and possibly acute coronary syndrome (ACS) presentation. Despite substantial knowledge about the clinical and technical aspects of SAT, the economic impact of these events has not been previously reported. Methods. We retrospectively reviewed 26 cases of SAT that occurred at our institution from 1998-2000. Baseline clinical and procedural data, as well as clinical outcomes for the initial hospital admissions required to treat SAT, were obtained by record review. Direct health care costs for these admissions were calculated based on hospital billing data and measured resource utilization for catheterization laboratory procedures. Results. Most cases of SAT occurred in high-risk circumstances, including ACSs, multi-stent interventions, and treatment of 3.0 mm or smaller vessels. Most patients suffered significant MIs and were treated with repeat percutaneous coronary intervention. The median time to SAT was 3.5 days, with 58% of events occurring on an outpatient basis. Median total hospital costs were $11,100 per patient, with more than half of the costs generated by the catheterization laboratory and pharmacy. Conclusions. Despite its low overall frequency, the clinical and economic costs of SAT are substantial. Specific strategies at preventing its occurrence are warranted, particularly in high-risk situations. PMID- 12082188 TI - Preventing subacute thrombosis. PMID- 12082189 TI - Conventional versus direct stenting in AMI: effect on immediate coronary blood flow. AB - There are no data regarding the potential benefits of direct stenting in the setting of AMI. The aim of this study was to evaluate the impact of direct stenting on the angiographic results and compare it to conventional stenting performed in the setting of AMI. We reviewed our institutional interventional database and identified 44 patients who had undergone stenting in the setting of AMI (29 primary and 15 rescue angioplasty). Patients were then divided into two groups; group A consisted of patients who had undergone conventional stenting (23 patients) and group B those who had undergone direct stenting (21 patients). Angiographic success was defined as TIMI flow grade 2. The baseline TIMI 0-1 flow was higher in group A compared to group B (74% vs. 24%; p < 0.05). TIMI flow rates before stenting (after balloon predilation in group A and after guidewire crossing in group B) and angiographic success (TIMI flow 2) after stenting were similar in both groups (p > 0.05). However, the final TIMI 3 flow were significantly better in group B after stenting (65% vs. 95%; p < 0.05). Although there was no no re-flow in group B, three patients in group A had no re-flow after balloon predilatation of lesions with baseline TIMI 2 flow. There was a statistical tendency to a higher TIMI 3 flow in patients treated with direct stenting in the setting of AMI. Direct stenting strategy in thrombus containing lesions seems to be a safe and feasible approach in avoiding no re-flow. PMID- 12082190 TI - A new miniature catheter with side-holes for percutaneous transradial or transbrachial coronary angiography. AB - The percutaneous arm approach through the radial or brachial artery for diagnostic cardiac catheterization has advantages, such as a lower incidence of access-site complications and decreased patient discomfort, particularly when smaller diameter catheters are employed. However, the pressure produced by high flow jets of contrast material exiting from an end-hole against the vascular wall can cause coronary dissection or myocardial blushing. To avoid this type of complication, we designed and developed a new miniature (4 French) catheter with two side-holes for coronary angiography. Under conditions similar to those of a clinical situation, we obtained the relationship between the pressure produced by the jets exiting from an end-hole and the force with which the tip of the catheter was pushed against the vascular wall. The presence of the side-holes substantially decreased the pressure at forces ranging between 0 and 15 gf, and at all injection rates (2.0 3.5 ml/second), but their location did not affect the pressure. In a preliminary clinical study, the new catheters with side-holes were feasible and effective for coronary angiography through the radial artery. Furthermore, the distribution of contrast material decreased the incidence of dislodgement of the catheter from the coronary ostia and enabled good visualization of the coronary ostia and proximal branches. PMID- 12082191 TI - Diagnostic angiography and catheter design. PMID- 12082192 TI - Transradial approach for renal percutaneous transluminal angioplasty and stenting: a feasibility pilot study. AB - BACKGROUND: Stent implantation of renal stenosis (RS) has been reported on extensively; this technique is perceived as superior to balloon angioplasty alone and less invasive than surgical revascularization. With the development of new accessories and stents, we can now improve and optimize procedural and clinical results. In this pilot study, we tested an alternative minimally invasive approach for renal stenting: the transradial artery catheterization (TAC) and a 6 French (Fr) system for stenting. METHODS: In this feasibility study, we enrolled patients with RS uncontrolled hypertension and/or chronic renal insufficiency. After assuring good dual blood supply (Allen test), TAC was attempted. The guiding catheters chosen for stenting were 6 Fr (0.70 ) Zuma 2 (Medtronic AVE, Santa Rosa, California). The selection of the guiding catheter was performed with respect to back-up support and coaxial alignment. RS were crossed with a coronary extra support guidewire (0.0.14 mailman, Boston Scientific/Scimed, Maple Grove, Minnesota). A balloon angioplasty before stenting was performed only in presence of subocclusive calcified stenosis, otherwise direct stenting was performed. We used low profile stents (12 16 mm LP Sulzer stent, IntraTherapeutics Inc., Minnesota) manually crimped on monorail balloon (5.0 6.0 mm x 20 mm). Immediately after completion of the procedure, the sheath was removed and a selective radial artery compression device was used for hemostasis. Patients were not restricted to bedrest, but were discharged the following day. RESULTS: There were 25 patients in the study population. Procedural indications were poorly controlled hypertension in 15/25 patients and/or preservation of renal function in 10/25 patients. Procedural success was obtained in 27/27 lesions (100%). Direct stenting was performed in 24/27 stenosis; predilatation was necessary in 3/27. We used 1 stent per lesion. The mean fluoro time was 7 5 minutes/lesion and mean procedural time was 14 5 minutes/lesion. No procedural and clinical complications occurred during the procedures upon the discharge. CONCLUSIONS: Our results confirmed the safety of this approach in the treatment of RS. The devices used in the study were those traditionally used for coronary procedures. The adoption of these materials and techniques have simplified the procedure and has permitted a reduction of fluoro and procedural time. An additional advantage has been the passive achievement of hemostasis by pressure devices reducing the workload of nursing and medical staff with elimination of bleeding complications that allowed immediate post procedural ambulation. PMID- 12082193 TI - Percutaneous revascularization of surgically corrected coarctation with graft restenosis. PMID- 12082194 TI - Platelet glycoprotein IIb/IIIa inhibitor use during percutaneous coronary intervention: IIb or Not IIb, what is the question? AB - Over the past decade, numerous placebo-controlled randomized clinical trials have documented robust clinical benefits of intravenous platelet glycoprotein (GP) IIb/IIIa inhibitors in patients undergoing percutaneous coronary intervention (PCI). This evidence has led to U.S. Food and Drug Administration approval and indication for use of two GP IIb/IIIa inhibitors at the time of PCI, namely, the chimeric monoclonal antibody fragment abciximab (ReoPro, Centocor, Inc. and Eli Lilly & Company) and the cyclic heptapeptide small molecule eptifibatide (Integrilin, COR Therapeutics and Key Pharmaceuticals). Currently, another small molecule GP IIb/IIIa inhibitor, tirofiban (Aggrastat, Merck & Company), which (similar to eptifibatide) is approved for the medical therapy of patients with non-ST segment elevation acute coronary syndromes (ACS), has not received indication for use in the PCI setting. Although the clinical benefits of both abciximab and eptifibatide administered at the time of PCI have been proven in randomized clinical trials, only abciximab has demonstrated a late survival advantage in patients following PCI. Evidence in support of the presence, magnitude and possible mechanisms for abciximab survival advantage is herein reviewed. PMID- 12082195 TI - Renal artery stenosis associated with saccular aneurysm and arterio-venous fistula. AB - A 25-year-old female presented with uncontrolled hypertension due to renal artery stenosis, a saccular aneurysm at the mid-shaft of the right renal artery, and an arterio-venous fistula distal to the aneurysm. Stent-graft implantation via the percutaneous approach was successfully used to treat this complex pathology. PMID- 12082196 TI - Successful stent treatment of pulmonary vein stenosis following atrial fibrillation radiofrequency ablation. AB - Pulmonary vein stenosis is a known complication of radiofrequency ablation of atrial fibrillation foci. We report a case of severe unilateral pulmonary vein stenosis following radiofrequency ablation resulting in no perfusion to the right lung. Simple balloon dilation was unsuccessful in this patient. Stent placement in the stenotic right pulmonary veins resulted in reperfusion of the lung as documented by follow-up lung scan at one year. PMID- 12082197 TI - Direct coronary stenting through left and right internal mammary artery grafts. AB - We describe a case of successful direct coronary stenting of two tight lesions, one at the site of the left internal mammary artery (LIMA) graft anastomosis with left anterior descending coronary artery and the other at the site of the anastomosis between the right internal mammary artery (RIMA) graft and the right coronary artery. To our knowledge, this is the first reported case of successful direct stent implantation through the LIMA and RIMA. PMID- 12082198 TI - Use of the X-SIXER catheter system in the treatment of acute thrombotic coronary occlusion. AB - The X-SIZER thromboatherectomy catheter system (EndiCOR Medical, Inc.) was initially developed for the treatment of thrombus in acute coronary syndromes. We present the case of a 64-year-old man with thrombotic occlusion of the left circumflex coronary artery. Antegrade flow was restored by aspiration of platelet thrombus at the point of occlusion, although the catheter was unable to cross the underlying stenosis. Its limited ability to cross severe stenoses or reduce lesion severity suggests that the X-SIZER should not be regarded as an atherectomy device. However, this case report suggests that the X-SIZER device has potential as an adjunct to PTCA and stenting in reducing the risk associated with thrombotic occlusion. PMID- 12082199 TI - Bleeding complications of platelet glycoprotein IIb/IIIa inhibitor abciximab (ReoPro ). AB - Studies of patients scheduled for percutaneous coronary intervention with acute coronary syndrome have shown that the addition of intravenous glycoprotein (GP) IIb/IIIa inhibitors to aspirin and heparin is associated with a reduction in death or myocardial infarction compared to therapy with aspirin and heparin alone. The principle safety issue with GP IIb/IIIa inhibitors is the risk of bleeding, as the potent antiplatelet effect of these drugs may adversely affect hemostasis. In addition, antagonists of GP IIb/IIIa may increase the risk of thrombocytopenia. We report a case of abciximab-induced severe thrombocytopenia which led to fatal intra-cranial hemorrhage. PMID- 12082200 TI - Thrombotic occlusion of the left main coronary artery during coronary angiography. AB - A case of left main occlusion due to thrombus during coronary angiography is reported. This rare and extremely dangerous complication was successfully managed with resuscitation maneuvers, hemodynamic support, urgent balloon angioplasty plus stent implantation, anticoagulation with heparin and potent therapy directed at preventing platelet aggregation. The thrombus most likely detached from the arterial sheath, although coexisting circumstances, such as the use of non-ionic contrast or a transient hypercoagulable state, may have had an influence. A careful technique during the procedure is probably the most effective preventive measure. PMID- 12082201 TI - A comparison of a single-stage isoproterenol tilt table test protocol with conventional two-staged tilt protocol in patients with syncope. AB - PURPOSE: Recently, Shen and colleagues described a single-stage isoproterenol protocol: Head Up Tilt Table testing (HUTT). The purpose of this study was to compare the introduction of this single-stage tilt protocol to sequential two staged tilt table testing. METHODS: Patients with recurrent or dramatic syncope or near syncope were referred for HUTT. A total of 100 patients underwent HUTT via a two-staged tilt protocol: 60 passive tilt for 30 minutes and, if negative, followed by a 10-minute tilt with isoproterenol titration to a 20% heart rate increase (two-staged HUTT). The next 100 patients underwent single stage isoproterenol protocol, which consisted of a 10-minute, 70 tilt with fixed dose isoproterenol infusion (0.05 ug/kg/minute). Patients with isoproterenol contraindications underwent a passive tilt (40 minutes for a conventional two staged 60-degree tilt, and 45 minutes for a single-staged 70 tilt. RESULTS: In the single-stage method, 40% of those that received isoproterenol had a positive test, as compared to 24% for two-stage HUTT protocol (p < 0.001). CONCLUSION: A single-staged isoproterenol HUTT provides a rapid method of increasing the overall test yield as compared to conventional two-staged HUTT. Further research is necessary to elucidate the overall utility of this rapid single-stage protocol. PMID- 12082202 TI - Clinical trials of intracoronary gamma radiation therapy for in-stent restenosis. AB - The only gamma emitter used in clinical trials for in-stent restenosis is 192Iridium (192Ir). The efficacy of intracoronary gamma radiation therapy in reducing clinical and angiographic restenosis in patients with in-stent restenosis has been established. This review is intended to give an overview of the clinical trials utilizing gamma vascular brachytherapy in patients with in stent restenosis and give insight into the future of intracoronary radiation therapy. PMID- 12082204 TI - Whose choice is it anyway? Decision making, control and conception. AB - Contemporary societies in the developed world are characterized by the expectation that women wish to and are able to control their experiences of fertility. Changes in medical technology and advances in reproductive medicine have played an important role in strengthening this expectation, together with other changes, such as the availability of free contraception. However, this article draws on data from two qualitative sociological research projects which demonstrate that women's expectations of reproductive choice and control are not always realized. Women's experiences of fertility are mediated by a dominant discourse which assumes that women both want and will achieve biological motherhood. The data indicate that women actively 'try' to achieve conception with the expectation that this is both controllable and easy. In some instances this seems to be the case, but many women soon realize that achieving conception is sometimes problematic or, indeed, impossible. This article concludes by arguing that although women may wish to control their experiences of fertility, their expectations of choice and control are frequently an illusion. PMID- 12082205 TI - Can fertility service providers justify discrimination against lesbians? AB - In Britain, for the last 25 years, lesbians have been seeking access to fertility services to achieve pregnancy. The problem for this particular group of clients is that often they are refused access to fertility services because of their sexuality. The reasons for treating lesbians less favourably than heterosexual clients are twofold: (1) donor insemination is defined as a treatment for infertility; and (2) service providers have a legal requirement to take into account the child's need for a father, according to Section 13(5) of the Human Fertilisation and Embryology Act 1990. The question addressed in this paper is whether these explanations constitute objective and reasonable justifications for discriminating against lesbians. As long as donor insemination is defined and perceived as a medical treatment for infertility, the social grounds for offering this procedure are obscured and there is apparent justification for treating lesbians less favourably. Once donor insemination is taken out of its medical context and understood in its social context, then it is no longer possible to justify discrimination. The 'need for a father' is the most common justification for discriminating against lesbians. Yet this phrase is ambiguous to the point of meaninglessness. Clearly, every child needs a father for conception to occur. None of the assisted conception techniques currently in use tamper with that basic need. This paper argues that commonly held beliefs about what a child needs from a father do not stand up to close examination. There is no evidence that a child needs a father for any of the following reasons: normal development, another parent, male role model, development of heterosexuality or social acceptance. If discrimination against lesbians were considered acceptable in other fields of family policy, it might legitimate discrimination in the field of reproductive medicine. However, the opposite is the case. In the fields of psychology, social work, adoption and family law, the evidence about the influence of parental sexuality on the welfare of the child is well known and frequently informs policy and practice. This paper defends the position that there are no objective and reasonable grounds to deny fertility services to lesbians. PMID- 12082206 TI - When (and what) is an egg? AB - The meaning of the term 'egg' is defined by the particular 'universe of discourse' in use. The discourse within an in vitro fertilization clinic will differ from the discourse in the domestic world of patients. Thus, it is imperative that any decision to donate or share eggs takes note of all possible universes, both present and future. PMID- 12082207 TI - The clinical value of sperm nuclear DNA assessment. AB - Traditional semen analysis is essential for the diagnosis of male infertility. A number of studies over the past decade have reported that a significant contributing factor to male fertility that is not revealed as part of semen analysis is sperm DNA, specifically its composition and organization. Exogenous and endogenous factors can cause damage to sperm DNA. For example, topoisomerase II activity, which is necessary for sperm DNA packaging, can adversely influence the competence of sperm DNA if the activity of the enzyme is abnormal. Germ cell apoptosis can be induced by oxygen radicals produced from environmental (for example cigarette smoke) or testicular (for example localized ischaemia) sources. Several assays have been developed that are useful for assessing sperm DNA composition and organization. To date, each of these assays has revealed that when sperm DNA has been damaged or packaged improperly there is a concomitant and often significant decline in male fertility. PMID- 12082209 TI - Radiation damage to the uterus -- review of the effects of treatment of childhood cancer. AB - At the present time approximately 1 in 1000 young people aged between 16 and 35 years will have been cured of cancer in childhood and some of the treatment regimens used will have predictable effects on their future fertility prospects. In young women who have been exposed to radiotherapy below the diaphragm, the reproductive problems include the risk of ovarian failure and significantly impaired development of the uterus. The magnitude of the risk is related to the radiation field, total dose and fractionation schedule. Premature labour and low birth weight infants have been reported after flank abdominal radiotherapy. Female long-term survivors treated with total body irradiation and marrow transplantation are also at risk of ovarian follicular depletion and impaired uterine growth and blood flow, and of early pregnancy loss and premature labour if pregnancy is achieved. Despite standard oestrogen replacement, the uterus of these young girls is often reduced to 40% of normal adult size. Uterine volume correlates with the age at which radiation was received. Regrettably, it is likely that radiation damage to the uterine musculature and vasculature adversely affects prospects for pregnancy in these women. It has been demonstrated that, in women treated with total body irradiation, sex steroid replacement in physiological doses significantly increases uterine volume and endometrial thickness, as well as re-establishing uterine blood flow. However, it is not known whether standard regimens of oestrogen replacement therapy are sufficient to facilitate uterine growth in adolescent women treated with total body irradiation in childhood. Even if the uterus is able to respond to exogenous sex steroid stimulation, and appropriate assisted reproductive technologies are available, a successful pregnancy outcome is by no means ensured. The uterine factor remains a concern and women who are survivors of childhood cancer and their carers must recognize that these pregnancies will be at high risk. PMID- 12082208 TI - Effect of inflammatory mediators on the physiology of the human Fallopian tube. AB - The physiological basis for the use of non-steroidal anti-inflammatory drugs (NSAIDs) in the treatment of acute pelvic pain in women was examined in terms of: (i) the electrophysiological responses of epithelial cells in the human Fallopian tube; and (ii) the longitudinal and circular contractions of the myosalpinx. Epithelial cells were grown as a polarized layer in primary culture, and transepithelial potential difference (p.d.) and short-circuit current (I(scc)) were recorded using a modified Ussing chamber. The inflammatory mediators histamine (0.1-100.0 micromol l(-1)) and platelet activating factor (PAF) (1.9 1900.0 nmol l(-1)) increased p.d. and I(scc) in a dose-dependent manner. Pre incubation with the NSAID diclofenac sodium (100 micromol l(-1)) inhibited the histamine- and PAF-induced stimulation of p.d. and I(scc). Aspirin (100 micromol l(-1)), ibuprofen (100 micromol l(-1)), indomethacin (100 micromol l(-1)) or naproxen (100 micromol l(-1)) were only partially effective. Histamine (0.1 1000.0 micromol l(-1)) increased the frequency of contractions of longitudinal and circular smooth muscle in segments of Fallopian tube in vitro, in a dose dependent manner. Pre-incubation with diclofenac significantly reduced the histamine-induced stimulation of tubal smooth muscle contraction at the higher doses of histamine. The other NSAIDs had no effect. These data provide evidence that diclofenac downregulates acute inflammation in the human Fallopian tube and may be of use as an anti-inflammatory agent in the treatment of pelvic inflammatory disease. PMID- 12082210 TI - Polycystic ovary syndrome and fat distribution: the central issue? AB - This review discusses the role of central obesity in polycystic ovary syndrome. There is increasing evidence that intraabdominal or visceral fat is either causative or a very early effect of polycystic ovary syndrome. The clinical implications of this are discussed. The simplest technique for assessing visceral fat and monitoring treatment is to measure waist circumference rather than waist:hip ratio or body weight. Weight loss is an effective treatment even where there is only a very modest reduction. The key element in lifestyle modification is regular exercise. PMID- 12082211 TI - Robotically assisted gynaecological surgery. AB - Industry has used robots successfully for fine, delicate, repetitive tasks for decades. Recently, robots have been introduced into clinical medicine and specifically into the surgical suite. Voice algorithms have been developed that allow voice activation of some types of equipment in the operating room, such as the laparoscope or the light source. Advances in computer software have allowed a computer controller to translate a surgeon's movements from the handles located in a console to the robotic arms that hold the surgical instruments. This console is placed away from the surgical table. Clinical experience is limited and there are few published clinical trials. The initial trials have focused on laparoscopic microsuturing such as that performed during coronary bypass surgery or tubal anastomosis. Preliminary results have demonstrated that laparoscopic coronary bypass surgery with the internal mammary artery can be achieved. In gynaecological surgery, laparoscopic tubal reanastomosis can be performed using the same technique that has been used traditionally at laparotomy. Future clinical trials will assess whether other gynaecological procedures can be performed with robotic assistance. PMID- 12082212 TI - Human cloning 2001. AB - This review summaries human cloning from a clinical perspective. Natural human clones, that is, monozygotic twins, are increasing in the general community. Iatrogenic human clones have been produced for decades in infertile couples given fertility treatment such as ovulation induction. A clear distinction must be made between therapeutic cloning using embryonic stem cells and reproductive cloning attempts. Unlike the early clinical years of in vitro fertilization, with cloning there is no animal model that is safe and dependable. Until there is such a model, 'Dolly'-style human cloning is medically unacceptable. PMID- 12082213 TI - A two DoF finger for a biomechatronic artificial hand. AB - Current prosthetic hands are basically simple grippers with one or two degrees of freedom, which barely restore the capability of the thumb-index pinch. Although most amputees consider this performance as acceptable for usual tasks, there is ample room for improvement by exploiting recent progresses in mechatronics design and technology. We are developing a novel prosthetic hand featured by multiple degrees of freedom, tactile sensing capabilities, and distributed control. Our main goal is to pursue an integrated design approach in order to fulfill critical requirements such as cosmetics, controllability, low weight, low energy consumption and noiselessness. This approach can be synthesized by the definition "biomechatronic design", which means developing mechatronic systems inspired by living beings and able to work harmoniously with them. This paper describes the first implementation of one single finger of a future biomechatronic hand. The finger has a modular design, which allows to obtain hands with different degrees of freedom and grasping capabilities. Current developments include the implementation of a hand comprising three fingers (opposing thumb, index and middle) and an embedded controller. PMID- 12082214 TI - Mechanical design of a shape memory alloy actuated prosthetic hand. AB - This paper presents the mechanical design for a new five fingered, twenty degree of-freedom dexterous hand patterned after human anatomy and actuated by Shape Memory Alloy artificial muscles. Two experimental prototypes of a finger, one fabricated by traditional means and another fabricated by rapid prototyping techniques, are described and used to evaluate the design. An important aspect of the Rapid Prototype technique used here is that this multi-articulated hand will be fabricated in one step, without requiring assembly, while maintaining its desired mobility. The use of Shape Memory Alloy actuators combined with the rapid fabrication of the non-assembly type hand, reduce considerably its weight and fabrication time. Therefore, the focus of this paper is the mechanical design of a dexterous hand that combines Rapid Prototype techniques and smart actuators. The type of robotic hand described in this paper can be utilized for applications requiring low weight, compactness, and dexterity such as prosthetic devices, space and planetary exploration. PMID- 12082215 TI - Biomimetic actuators in prosthetic and rehabilitation applications. AB - Where humans and mechanical systems operate in close proximity there is a need to provide drive systems that combine the positive attributes of conventional actuator design with a 'softer' safer interaction capacity. This is achieved by natural muscle, and engineering emulation of this functionality could have a significant benefits in many areas, but particularly the medical domain. This work will study the use of compliance regulated and controlled pairs of antagonistic pneumatic Muscle Actuators (pMAs) in two medical scenarios; i) The construction of dexterous prosthetic hands having a high power and low mass potential, ii) The construction of a power assist device that can be used to augment the strength of those suffering from degenerative muscle wasting diseases. PMID- 12082216 TI - High torque ultrasonic motors for hand prosthetics: current status and trends. AB - This paper presents the work carried out at the Instituto de Automatica Industrial, in the framework of the EU MANUS project and with the co-operation of the Katholieke Universiteit of Leuven, to develop a specifically designed TWUM for driving our prototype of hand prosthesis. The application of TWUM to prosthetic devices has clear advanteges, namely, noiseless operation, self braking drive, high torque and low speed characteristics. Here we analyse main actuation advantages, point out main drawbacks of this technology and describe our research in the area of specific design of TWUM for high torque actuation, improved control characteristics and enhanced overall performance. We intend to provide a concurrent approach to the overall actuator design involving electronics, mechanical design as well as improved control strategies having in mind the prosthetic application. PMID- 12082217 TI - Generating pathological gait patterns via the use of robotic locomotion models. AB - In this article we explore the feasibility of modeling normal and pathological human gait using a relatively simple five-element model. We use a robust, nonlinear control scheme to regulate the gait patterns of the model. Simulated gait patterns are generated through the use of five constraint relationships that depend on four gait parameters. Two pathological conditions due to muscle weaknesses were simulated by modifying the control torques at the joints. We demonstrate that the model successfully approximates the qualitative and quantitative dynamic trends that were observed in normal and pathological human locomotion. PMID- 12082218 TI - A new optics-based gastroesophageal reflux probe. AB - This work was carried out with the purpose of developing a new method of gastroesophageal reflux (GOR) detection. It is based on the emission of a light beam to the inferior part of the oesophagus and on the detection and analysis of the corresponding reflected light intensity. The optical properties of the oesophageal lumen are then used to identify the GOR episodes, solving, in this way, the existing drawback of using pH probes that fail in the cases where GOR episodes are neutral or short duration acid. The necessary instrumentation for the application of this new technique, including the probe itself and its associated optics, was developed. The result is a low-cost portable instrument, based on the Microchip microcontroller PIC16C77, with enough flexibility to be used in other biomedical applications. This new simple apparatus only needs an adequate light source - diode laser - and an adequate photosensor - photodiode - to make the interface to the probe that guides the light to and from oesophagus. Our results show the capability of this new technique to make the identification of GOR episodes. PMID- 12082219 TI - Sprouting in the hippocampus after entorhinal cortex lesion is layer- specific but not translaminar: which molecules may be involved? AB - Entorhinal cortex lesion partially denervates the rat fascia dentata. This is said to induce sprouting of intact fibers from neighboring layers that invade the zone of the degenerating axons. However, recent in vivo and in vitro studies failed to demonstrate sprouting across laminar boundaries. Sprouting does occur, but it mainly involves unlesioned fiber systems terminating within the layer of fiber degeneration. These findings point to laminar cues that promote sprouting of fibers within the denervated zone while repelling other, adjacent fiber systems that try to grow into the denervated zone. A group of molecules that are likely to guide the sprouting process and the formation of borders are extracellular matrix molecules synthesized by reactive astrocytes. These molecules provide boundaries for growing axons during development. Some extracellular matrix molecules (tenascin-C, DSD- 1 -proteoglycan, neurocan, and brevican) were upregulated within the denervated outer molecular layer after lesion of the entorhinal cortex, suggesting a similar role after lesion. These extracellular matrix components forin a sharp molecular border towards the adjacent nondenervated inner molecular layer, and their pattern of distribution correlates precisely with the laminar termination pattern of the sprouting fiber populations. Thus, extracellular matrix molecules could delineate boundaries of axonal growth after entorhinal cortex lesion and could thus contribute to the molecular processes underlying the postlesional re-patterning of the fascia dentata. PMID- 12082220 TI - Developmental and reactive growth of dentate gyrus afferents: cellular and molecular interactions. AB - The lamination of dentate gyrus afferents established during development is maintained following lesion-induced reactive growth in the adult. After partial deafferentation sprouts from undamaged afferents restore most synapses, while respecting the laminae relative boundaries. No evidence of trans-laminar sprouting has been found. Here, we review the information gathered during the last decade on the cellular and molecular bases of dentate synaptogenesis, with special attention to the role of glia during development and that of reactive glia after deafferentation. The interactions of neurons with astroglia and astroglial macromolecules, particularly proteoglycans, influence synapse segregation in the dentate gyrus, providing us with a reasonable explanation for afferent lamination. PMID- 12082221 TI - Plasticity following lesion: help and harm from the immune system. AB - In contrast to other organs where the tissue is capable of replacing lost cells and thus regaining tissue function, immune cell recruitment and activation is suppressed in the CNS in order to minimize secondary damage after lesion. This state of immune privilege has its cost because the injured tissue cannot fully benefit from growth-promoting effects accompanying inflammatory responses. These responses include phagocytosis of growth-inhibiting myelin debris by cells of the innate immune system and the recently described protection of surviving fibers by myelin-specifie T cells of the adaptive immune system. While the signals suppressing macrophage functions in the CNS are yet to be defined, it seems that help from T cells is diminished by apoptosis-induction via death-inducing ligands. Indeed, the death ligand CD95L (FasL, APO 1 L) is constitutively found on neurons, microglia and astrocytes. Its upregulation on astrocytes during axonal degeneration in the hippocampus after entorhinal lesion is accompanied by the appearance of apoptotic leukocytes. T cells also express CD95L and TNF related apoptosis- inducing ligand (TRAIL), and the presence of CD95 (Fas, APOI) and TRAIL-receptors renders brain cells putative targets of T cell-induced apoptosis. Thus, blockade of death ligands could be helpful by simultaneously enhancing T cell survival and blocking T cell-mediated brain cell death. This is only one example of how boosting helpful immune cell functions and abrogating their destructive effects might help to overcome the brain's failure to regenerate after axonal lesions. PMID- 12082222 TI - Back-propagating action potentials in pyramidal neurons: a putative signaling mechanism for the induction of Hebbian synaptic plasticity. AB - A hallmark of synaptic plasticity is the associative, or Hebbian, nature of its induction. By associative, we mean that the timing relationships between activity of the pre- and postsynaptic elements of a synapse determine whether synaptic strengths are modified. lt is well-established that associativity results, in large part, from the dual requirements for activation of the N-methyl-D-aspartate receptor-ionophore, namely presynaptic neurotransmitter release and postsynaptic depolarization. However, the specific dendritic events that provide the postsynaptic depolarization have been relatively unexplored. Increasing evidence suggests that back-propagating (i.e., antidromic) Na(+) action potentials provide the necessary postsynaptic depolarization to allow induction of associative synaptic plasticities. In hippocampal CAI and neocortical layer V pyramidal neurons, these action potentials provide much greater levels of dendritic depolarization than would be expected from synaptic currents alone. Moreover, they provide a relatively brief and synchronous depolarization throughout the dendritic arbor, allowing timing relationships to more directly reflect pre- and postsynaptic cell firing. Interestingly, certain properties of the back propagating actions potentials differ from axonal or somatic action potentials in ways that seem to reflect their function. For example, the all-or-none property of action potential amplitude does not hold in the dendrites. In this review we discuss the back-propagating action potential as a dendritic signal that provides information to synapses about the firing state of the postsynaptic neuron. First, we consider the evidence that action potentials propagate back from the axon. Second, we describe the characteristics of the back-propagating action potential in terms of interactions of its underlying ionic currents. Third, we describe how these properties contribute to the timing aspects of the induction of long-term potentiation. Finally, we discuss modulation of the underlying ion channels by neurotransmitter systems and other agents and speculate on their roles in learning and memory. PMID- 12082223 TI - Interactive pathology following traumatic brain injury modifies hippocampal plasticity. AB - Hippocampal afferents terminate in well-defined laminae, with a morphological segregation of input which has facilitated the interpretation of structural and functional synaptic reorganization observed after deafferentiation. Historically, most studies have induced hippocampal plasticity using single deafferentiation paradigms, however recent evidence indicates that sequential lesions or models based on combined injuries alter the pattern of dendritic structural reorganization and axonal sprouting. A better understanding of the interaction between deafferentiation-induced structural remodeling and other pathological mechanisms, which commonly coexist in central nervous system trauma, will require the use of combined injury paradigms where such plasticity can be systematically manipulated. In the context of traumatic brain injury, we have developed an injury model that combines the excessive neuroexcitation of concussive brain insult with the targeted hippocampal deafferentation of entorhinal cortical lesion. This review discusses the role of such an approach in defining posttraumatic hippocampal vulnerability, out- lining the effects of combined pathology on hippocampal circuitry, and considers the greater clinical relevance inherent in the combined injury approach. Experimental evidence obtained with the combined concussive plus deafferentation model is presented, detailing the interaction of injury components and highlighting structural, behavioral and electrophysiological evidence for maladaptive hippocampal plasticity. Subsequent studies utilizing pharmacological methods to manipulate this maladaptive plasticity are described, first targeting glutamate, acetylcholine and dopamine receptor pathways, and then applying select drugs to explore how various molecular mechanisms underlying combined neuroexcitation and deafferentation pathology might affect regenerative plasticity. Evidence implicating postinjury neurotransmitter modulation of exeitatory/inhibitory homeostasis, metalloproteinase regulation of extracellular matrix, and mitochondrial metabolic vulnerability is presented. Finally, the effect of age on outcome after combined neuroexcitation plus deafferentation insult is considered, as well as how future studies in such combined injury models will better define the full range of postinjury hippocampal plasticity possible after brain trauma. PMID- 12082224 TI - The role of axonal sprouting in functional reorganization after CNS injury: lessons from the hippocampal formation. AB - Functional reorganization is often invoked to account for recovery of function after central nervous system (CNS) injury. The mechanisms underlying this possible reorganization, however, remain uncertain. In the last 30 years, studies of the hippocampal formation of rats have indicated that the CNS is capable of undergoing significant changes in its pattern of connectivity in response to injury. Here, we explore numerous examples of lesion-induced alterations in hippocampal connectivity known as axonal sprouting. Both homotypic and heterotypic sprouting occur in the denervated hippocampus after unilateral entorhinal cortex lesions. We assess the behavioral relevance of glutamatergic homotypic sprouting emerging from the surviving contralateral entorhinal area (i.e., the crossed temporodentate projection) as well as the heterotypic sprouting from the remaining surviving afferents (e. g., the cholinergic septodentate pathway) to the hippocampus. Studies examining the role of crossed temporodentate sprouting in recovery from memory deficits after entorhinal cortex injury indicate that homotypic sprouting may indeed contribute to a reorganization of cortical function resulting in recovered mnemonic capacity. Heterotypic sprouting is not as clearly linked to recovery of function after bilateral entorhinal injury. We propose a tripartite model for functional reorganization based on homotypic sprouting, neurotrophic factors, and altered inhibitory functioning to account for how relatively small increases in surviving homotypic pathways might restore neurological function. PMID- 12082225 TI - Gene expression profiling and molecular aspects in peripheral nerve regeneration. AB - Regeneration of the peripheral nervous system after injury depends on a complex sequence of histopathological reactions that comprise a highly reproducible sequence of degenerative reactions, termed Wallerian degeneration. During this period a remodelling of the distal nerve stump prepares a microenvironment that permits successful regrowth of nerve fibers from the proximal nerve fragment. This stereotypical sequence of reactions is reflected by a differential and coordinate expression of genes with specific functions in the process of regeneration. This review will summarize cellular and molecular reactions that contribute to peripheral nerve regeneration including data of a pilot study in which membrane based cDNA array expression technology was applied. We examined the expression of 588 annotated genes in response to a crush lesion of rat sciatic nerves. Approximately 40 % of the genes spotted onto the array filters showed expression significantly above background and 55 of these detected genes represented differential expression profiles after nerve lesion. This approach revealed to be suitable for systematic screening of regeneration associated genes. PMID- 12082226 TI - Degeneration of axotomized retinal ganglion cells as a model for neuronal apoptosis in the central nervous system - molecular death and survival pathways. AB - Programmed cell death (PCD) or apoptosis is a phenomenon important for proper development and morphological as well as functional fine tuning of the nervous system. In the past two decades it became evident that the same apoptotic machinery, which has crucial functions in during development, can be reactivated under pathological circumstances in the adult nervous system and contribute to neuronal cell loss due to various neurological disorders like ischemic stroke, neurodegenerative diseases or brain traumata. In this review, we present the optic nerve transection paradigm as a valuable model for investigation of apoptotic neuronal cell death in the central nervous system (CNS). We review and summarize the most important discoveries regarding molecular pathways and mechanisms of neuronal apoptosis during the past few years, and outline contributions that have been made investigating the death of retinal ganglion cells (RGCs) following transection of the optic nerve. PMID- 12082227 TI - The JNK and p38 signal transduction following axotomy. AB - The transection of nerve fibers evokes a characteristic reaction in the injured neurons, the so-called cell body response (CBR), which comprises aspects of developmental re-differentiation with parallel loss of the transmittory phenotype, efforts or achievement of axonal elongation and re-construction of effective synapses. Neither the signals underlying the onset of CBR nor the programs underlying regeneration are sufficiently elucidated. Here we review the putative role of two subfamilies of the MAP kinases, the JNKs (c-Jun N-terminal kinases) and the p38 kinases in the CBR. Following nerve injury with subsequent CBR, JNKs are rapidly activated and this activation persists for weeks until neu ronal cell death or successful regeneration. The various functions render JNKs to perfect candidate molecules for the realization of the CBR including axonal transport, activation of c-Jun, modulation of cytoskeletal functions, detection of cytoskeletal alterations, or signal transduction of adhesion molecules in the axon and growth cone. On the other hand, the rapid but transient activation of p38 might interfere with the mitotic arrest, a putative feature of the CBR. PMID- 12082228 TI - A purine-sensitive mechanism regulates the molecular program for axon growth. AB - Axon growth is characterized by a distinctive program of gene expression. We present evidence here that this program is regulated through a purine-sensitive mechanism, and that it can be re-activated in mature CNS neurons to induce extensive axon growth in vitro and in vivo. In dissociated goldfish retinal ganglion cells, the purine nucleoside inosine acts intracellularly to stimulate axon outgrowth by inducing the expression of GAP-43, Talpha-1 tubulin, and other growth-associated proteins. The purine analog 6-thioguanine (6-TG) acts in the opposite fashion, blocking axon growth and the underlying program of molecular changes. Prior studies in PC12 cells have shown that 6-TG selectively inhibits the activity of N-kinase, a 47-49 kDa serine-threonine kinase. Inosine acts as a competitor of 6-TG, suggesting that it acts as an N-kinase agonist, and that this kinase is part of a modular signal transduction pathway controlling axon growth. Following unilateral transections of the corticospinal tract in mature rats, inosine applied to the intact sensorimotor cortex stimulated layer 5 pyramidal cells to upregulate GAP-43 expression and to sprout axon collaterals that crossed the midline and reinnervated regions of the cervical spinal cord which had lost their normal afferents. It will now be important to identify the molecular changes that lie upstream and downstream of N-kinase, and to explore the clinical potential of activating this pathway in patients who have sustained CNS injury. PMID- 12082229 TI - Tenascin glycoproteins and the complementary ligand DSD-1-PG/ phosphacan- structuring the neural extracellular matrix during development and repair. AB - The differentiation and morphogenesis of neural tissues involves a diversity of interactions between neural cells and their environment. Many potentially important interactions occur with the extracellular matrix (ECM), a complex association of extracellular molecules organised into aggregates and polymers. The large modular glycoprotein, Tenascin-C, and the chondroitin sulphate proteoglycan, DSD-1-PG/Phosphacan, have complex and frequently overlapping expression patterns in the developing CNS. Their presence in zones of cell proliferation, migration, and differentiation, as well as in boundary structures, suggest that they may be involved in the modulation of an extensive range of cellular processes. They are both strongly up-regulated in a range of CNS lesions and pathologies, being components of the glial scar, and expressed by gliomas. Functional roles in many cellular processes are possible through their extensive molecular interaction sites, both with each other, and with many of the same cell surface receptors, adhesion molecules, growth factors and other matrix proteins. These multiple interactions involve sites on both their protein domains and on the heterogeneous carbohydrate groups with which they are post-translationally modified. In vitro assays demonstrate cell-type specific effects on adhesion, migration and the formation and extension of cellular processes, including neurites and axons. PMID- 12082230 TI - The influence of astrocytes on the development, regeneration and reconstruction of the nigrostriatal dopamine system. AB - Over the past few decades astrocytes have emerged from being considered simple packing tissue in the brain to become major players in the development, survival and functioning of central nervous system (CNS) neurons. As the influence that astrocytes (and the various molecules they produce) have on the development of CNS neurons becomes more evident, it will be important to consider how this information can be exploited to bring about better protection, recovery and/or regeneration of circuits which are destroyed in the adult CNS due to trauma or com-mon neurodegenerative episodes. Although the characterisation of astrocytic responses to brain injuries, neurodegenerative disease, and cell transplantation are becoming more common, we still known little about how astrocytes influence the (re)growth or reconstruction of neural circuitry after the development period is ended, or indeed what is the overall impact of an astrocytic presence on the growth of neurons in the adult CNS. With the major hurdle of recognition of the importance astrocytes in the function and recovery of the adult CNS now cleared, a new chapter in the development of powerful new treatments for CNS disorders and injuries is now open. The following is a brief review of what we know about how astrocytes influence the growth and connectivity of the nigrostriatal circuit during development, and how these cells may affect efforts to reform this circuit after it s destruction/degeneration in the adult CNS (as commonly happens in Parkinson s disease). As we obtain more information on the specific influence of these cells in various developmental, traumatic and disease events we can expect to find better ways toward combating major disorders of the human CNS. PMID- 12082231 TI - Regenerative events in the olivocerebellar pathway. AB - Mature neurons display a wide range of regenerative capabilities. As a general rule, peripheral neurons have the highest regenerative abilities both in the form of terminal sprouting and of axonal elongation following axotomy, regardless of the distance of the lesion from the cell body. In contrast, in central neurons reactive sprouting has been demonstrated in a limited number of neuronal populations and this type of growth may be dependent on the constitutive presence of specific growth-associated proteins. Central axon elongation is critically dependent on the presence of suitable environment and on the intrinsic capabilities of each neuronal population. These capabilities are controlled at least in part by repressive signals that are mainly located along the axons. They are more easily disclosed when a short axon stump is left after axotomy. The adult olivary neurons offer a unique model in the central nervous system for their remarkable plastic properties: i) they undergo extensive remodeling of their terminal arborizations following target manipulations or under the influence of electrical activity; ii) they are capable of axonal regeneration in a suitable environment; iii) their response to injury does not depend on the distance of the axotomy from the cell body. In this respect they are similar to peripheral neurons and likely their target cells are the main source of the repressive signals control-ling growth genes. The demonstration that this pathway is also able to find the proper target cells provides a striking example of how the mature brain may be repaired through appropriate manipulations. PMID- 12082232 TI - Regeneration in the mammalian optic nerve. AB - Since the first studies on axonal regeneration, the optic nerve (ON) of higher vertebrates has been considered a good experimental system to investigate the failure of mature CNS neurons to re-grow after axotomy. The optic nerve is composed of a single population of fibers the RGC axons and, being separated from the rest of the brain, it is easily accessible to surgical manipulations. All the fibers can be transected without massive damage to the surrounding tissue, so their reaction to axotomy is not perturbed by extended inflammation processes. Another advantage of the system is the accessibility of RGCs. Being in the more internal retinal layer, RGCs are directly exposed to the humor vitreus, the liquid filling the posterior chamber of the eye. Pharmaceutical treatments are easily injected into the eye and, diffusing in the vitreus, can reach all the RGCs. Last but not least, functional recovery can be easily monitored in the optic nerve; measurement of electrical activity in response to visual stimuli in CNS regions that receive inputs from the retina such as superior colliculus or visual cortex allows evaluation of the re-growth of ON fibers and the restoration of connections. All the experiments carried out so far indicate that the failure of regeneration in the ON, as in the majority of the CNS districts, is a multi-factorial phenomenon, involving three classes of negative events. 1) RGCs die after axotomy: in the adult rat, their number is reduced to a very small percentage in a few weeks after the lesion. 2) The majority of mature axotomised RGCs are not programmed to re-start the process of axonal elongation that they displayed in immature stages. 3) The optic nerve environment contains molecules many of them upregulated after the lesion that are inhibitory for axonal growth. This review, focused on experiments performed in the mammalian optic nerve, traces attempts made to overcome each of these three obstacles, and maps progress towards a combined therapeutic strategy. PMID- 12082233 TI - Neural transplantation: restoring complex circuitry in the striatum. AB - During the last 30 years, the promise of neural transplantation as a therapeutic strategy for neurodegenerative disease has been slowly recognised. Across the world, clinical transplants of embryonic primary dopamine neurones have been shown to ameliorate some of the motor deficits in Parkinson s disease (PD) patients, and more recently, systematic clinical trials have been initiated for the replacement of striatal projection neurones lost in Huntington's disease (HD). Clinical transplantation as a prospective therapy for HD poses a particular set of difficulties. The hallmarks of this neurodegenerative disease include extensive loss of medium spiny long-distance projection neurones of the caudate and putamen, affecting downstream target nuclei, the globus pallidus and substantia nigra, leading to dysregulation of motor control. In addition, extensive loss of cortical neurones that form the afferent systems to the basal ganglia leads to widespread cognitive decline. If transplantation therapy is to succeed in replacing degenerating neurones in HD and reinstating controlled function of complex basal gan-glia circuitry, the new neurones must be able to develop specific long-distance projections that can form accurate and functional connections with neurones in precise target regions. Our ongoing studies are aimed at addressing how we can improve the function of striatal transplants, in particular to optimise the reformation of precise long-distance connections and to re-establish normal motor and cognitive function. In particular, we have investigated optimal requirements for embryonic primary tissue to achieve these aims, and also the potential of other cell sources to provide long-distance projection neurones and reconnect complex circuitry. This review describes current progress of experiments to optimise the reconstruction of neuronal circuitry using primary embryonic tissue transplants, as well as our current initiatives to use neural stem cells or precursors to replace long distance projection neurones in the degenerating basal ganglia. PMID- 12082234 TI - The collagenous lesion scar--an obstacle for axonal regeneration in brain and spinal cord injury. AB - After CNS trauma a sheet-like, collagen type IV (Coll IV) immunopositive basement membrane (BM) develops in the lesion zone as well as at newly formed blood vessels. The basic scaffold of this BM is composed of Coll IV, laminin and nidogen but numerous other proteins some of which are discussed to be inhibitory for axonal regeneration, i.e. chondroitin- and heparansufate-proteoglycans, are associated with BM. This review will focus on the collagenous wound healing scar, discuss its composition and summarize the experimental results that demonstrate its role in the failure of axonal regeneration in the injured mammalian CNS. PMID- 12082235 TI - Cell therapy to repair injured spinal cords: olfactory ensheathing glia transplantation. AB - The absence of spontaneous axonal regeneration in the adult mammalian central nervous system cause devastating functional consequences in patients with spinal cord injuries. During the past decades several attempts have been made in order to find a strategy to repair injured spinal cords in experimental animals, that could provide a novel therapeutic approach in humans. Cell transplantation has been broadly used as an intervention to influence neuronal survival and axonal regeneration in the severed neuraxis. Of the cell types used for transplantation, olfactory ensheathing glia (OEG) promoted a dramatic functional improvement and anatomical repair after complete transection of the adult mammalian spinal cord. These cells can be easily obtained from adult donors opening the possibility of autologous transplantation. Grafting OEG to repair injured spinal cords offers some advantages compared to injections of other cell types. Therefore, OEG have become good candidates to bring about repair in damaged spinal cords. In this article we review OEG transplantation studies, discuss the properties that could account for their axonal growth-promoting ability, and the advantages of using OEG as a repair strategy. PMID- 12082236 TI - Advances and issues in the diagnostic differential of malingering versus brain injury. AB - The past decade has seen numerous advancements in the assessment of malingered brain injury, though the current diagnostic system offers only guidelines in which malingering should be suspected. This article presents an overview of advances in the clinical and neuropsychological assessment of malingering, issues in diagnostic differential, neuropsychological test methods, and special issues presented by the medical-legal context, and other factors which may affect presentations. Cautions and recommendations for practice are presented. PMID- 12082237 TI - The phenomenology of depression after brain injury. AB - One important challenge in neuropsychiatry is how to diagnose depression in patients with acute brain lesions, since there may be an overlap between symptoms of depression and signs associated with the neurologic disease. The best approach is to assess the presence of depressive symptoms using semi-structured or structured psychiatric interviews such as the Present State Exam, the Structured Clinical Interview for DSM-IV, or the Schedules for Clinical Assessment in Neuropsychiatry. The diagnosis of a depressive syndrome should be made using standardized diagnostic criteria for mood disorders due to neurological disease such as in the DSM-IV or the ICD-10. Depression rating scales, such as the Hamilton Depression Scale and the Center for Epidemiologic Scales for Depression may be used to rate the severity of depression and monitor the progression of antidepressant treatment. Most studies in acute and chronic neurologic disorders demonstrated the specificity of both autonomic and psychological symptoms for the syndrome of depression. The present review article examines important considerations before a diagnosis of depression in neurologic disease, discusses a variety of psychiatric instruments that are used to examine the presence and severity of depression in neurologic disease, examines relevant phenomenological issues, and proposes different diagnostic strategies. PMID- 12082238 TI - Dementia Due to Head Trauma: An obscure name for a clear neurocognitive syndrome. AB - This paper questions the DSM IV TR criteria for Dementia Due to Head Trauma (DDHT). We studied 20 consecutive traumatic brain injury (TBI) patients and checked them for this criteria. We found the diagnosis criteria to be oversensitive and lacking in specificity, consequently raising the possibility for all subjects who have sustained severe TBI to be diagnosed with dementia. A careful analysis of data and literature showed that nearly half of these patients were able to return to work after undergoing an intensive and holistic rehabilitation program, indicating a reversal of the "dementia". Severe head injury produces a set of multiple cognitive deficits that shouldn't be considered dementia. The term "Severe Neurocognitive Disorder" is suggested for use instead of DDHT for the cognitive deterioration of severe TBI patients. PMID- 12082239 TI - DSM-IV: Diagnosis of children with traumatic brain injury. AB - This paper reviews the strengths and limitations of the DSM-IV in the diagnosis of children with Traumatic Brain Injury. The authors compare the DSM-IV diagnostic characteristics for effective classification and decision making in children with TBI. The DSM IV multiaxial approach can be useful in patients with brain injury because medical considerations can be incorporated with the diagnosis, along with standard mental health factors. PMID- 12082240 TI - Recommendations for the revision of DSM-IV diagnostic categories for co-morbid posttraumatic stress disorder and traumatic brain injury. AB - The definitions in the Diagnostic and Statistical Manual-4th Ed.-TR [5] relating to posttraumatic stress disorder (PTSD) and traumatic brain injury (TBI) after mechanical accidents do not reflect the range of dysfunctions, the significance of patient differences, and fluctuating intensity, direction, and symptoms with increasing time since an injury. The considerable overlapping of symptoms is not considered at all, nor is concussion given a diagnosis. Some anatomical and physiological considerations for these frequently comorbid conditions are specified to increase diagnostic precision, or call attention to unavoidable ambiguities. The current definitions lead to diagnostic error since there are many that are based upon symptoms that are not trauma related although they resemble those of TBI. A Taxonomy of Neurobehavioral Disorders draws attention to a wide range of physiological and behavioral functions, with implications for more accurate recognition of symptoms, diagnosis, and treatment. Persistent posttraumatic disorders, more complex than PTSD, exist, due to the effects of unhealed tissues, impairment, and the social consequences of impairment and rejection. There are recommendations for the improvement of the definitions of PTSD and traumatic brain injury when both may be co-morbid after a mechanical injury. PMID- 12082241 TI - Eye movements and verbal reports in neglect patients during a letter reading task. AB - OBJECTIVES: To study different typolologies from visuo-verbal behavior concerning the arrest and the treatment of visual information in patients with spatial hemineglect. METHODS: Ocular movements were monitored by photo-oculography in thirty controls and twenty-three patients with neglect. During a letter reading task, the movements, number of entries into predetermined zones of the visual field, time spent in each zone, and the verbal responses were recorded. Four profiles may be anticipated for such trials: normal scanning and reading (S+R+), normal scanning without reading (S+R-), absence of both scanning and reading (S-R ), abnormal scanning with reading (S-R+). RESULTS: The letter projected in each of the 4 quadrants was scanned and correctly identified by all of the controls with no differences in scanning times. Among patients, the 4 possible behavior patterns were found along with differences in exploration duration per zone (p < 0.05) and number of times specific zones were attained (p < 0.05). CONCLUSIONS: Our results confirmed the heterogeneity of the mechanisms of neglect and suggest that rehabilitation procedures adapted to each profile might be useful. PMID- 12082242 TI - Recovery of proximal and distal arm weakness in the ipsilateral upper limb after stroke. AB - Conservation of the ipsilateral upper limb function is important in stroke subjects with contralateral hemiplegia, because often it must serve as a compensatory tool for activities of daily livings (ADLs). However, the amount of functional loss and/or the recovery pattern of the ipsilateral upper limb are not well known. We plan to investigate how to measure the progress of the ipsilateral upper limb function after the onset of stroke. Once a week we used the Manual Function Test (MFT) to evaluate the ipsilateral upper limb weakness of 72 stroke subjects with contralateral hemiplegia until maximum recovery. Proximal and distal arm weaknesses in the ipsilateral upper limb were maximally recovered within one month following the onset of hemispheric stroke, but their weakness was not to be completely recovered. Also the amounts of their recoveries were different from each other. These results indicate that the ipsilateral upper limb weakness in stroke is not a temporary event and that motor function of the proximal and distal arm might be mediated by different neuronal circuits. PMID- 12082244 TI - Internet resources for traumatic brain injury: A selective review of websites for consumers. AB - This article provides a selective review of brain injury resources found on the Internet. It is geared toward the needs of survivors of brain injury and their families. Provided are listings of websites that focus on brain injury, those with sections pertaining to brain injury, and those with brain injury information found by searching the site. Sites that afford communication in the form of chatrooms, message boards, and mailing lists are described and listed. The value of this technology for consumers, as well as some of the challenges in searching for relevant materials on this topic, is discussed. PMID- 12082243 TI - Aerobic exercise intervention improves aerobic capacity and movement initiation in Parkinson's disease patients. AB - Parkinson's Disease (PD) is a progressive neurologic disorder, which includes an inability to activate appropriate muscle activity. Very little research has analyzed aerobic exercise for PD patients. The purpose of this study was to investigate the effect of a 16 week aerobic exercise intervention on aerobic capacity and movement initiation (MI) time for PD patients. With 8 PD subjects (Hoehn & Yahr stage 2), 4 completed the exercise intervention. Peak VO_2scores significantly improved (26%) Choice MI improved from 532 ms to 415 ms, while simple MI improved from 285 ms to 261 ms. The improvement in aerobic capacity suggests that PD patients may benefit from exercise just as much as a normal population. The change in MI indicates that aerobic exercise may reduce the detrimental effects of neuromuscular slowing within PD patients, by improving the subjects' ability to initiate and perform appropriate movement patterns. PMID- 12082245 TI - Comparative analysis of aggregate shapes by digitized microscopic images. Application to hypertension. AB - The main objective of the present work was to study modifications in RBC aggregate morphology by analyzing digitized microscopic images and compare them between healthy subjects and patients suffering from essential hypertension. Blood samples were obtained from normal subjects (n=30) and patients suffering from essential hypertension (n=20). RBC aggregate morphology was quantified using direct microscopic observation and numerical analysis of images. ASP (Aggregation Shape Parameter) defined as the ratio of the area of the projected image to its square perimeter was calculated. Other rheological parameters have been determined in order to establish the hemorheological profile of the studied hypertension states. ASP appears significantly higher (p<0.001) in patients suffering essential hypertension (0.69+/-0.11) than in normal control subjects (0.25+/-0.12). RBC aggregation is known to be responsible for the high increase in apparent blood viscosity at low shear rates. By compare ASP values with whole blood viscosity at low rate (2.30 s(-1)) a high correlation was formed between both parameters (Spearman coefficient was 0.8835 and p<0.001). The applied method is simple, direct and quantitative and provides a useful tool for measuring deviations of RBC aggregate morphology. PMID- 12082246 TI - Hemorheology and hemostasis in vascular disease. A pathophysiological review. PMID- 12082247 TI - Exercise hemorheology as a three acts play with metabolic actors: is it of clinical relevance? AB - Hemorheological effects of exercise are a triphasic phenomenon including: (a) short-term effects (hyperviscosity mostly due to fluid shifts and alterations of erythrocyte rigidity and aggregability); (b) middle-term effects (i.e., the reversal of acute effects due to plasma volume expansion (autohemodilution) that lowers both plasma viscosity and hematocrit; (c) long-term effects that further improve blood fluidity, parallel with the classical training-induced hormonal and metabolic alterations. Red cell rheology during these 3 stages is affected by white cells and oxidant stress. On the other hand, most metabolic and hormonal alterations play a role in exercise-induced hemorheological changes: among them, blood lactate appears to have opposite effects according to the training status, since it generally impairs erythrocyte fluidity while it improves it in some subgroups of highly trained athletes, a difference that could be related to membrane monocarboxylate transporter status. Body composition (mostly hydration status and the amount of fat mass) as well as its major hormonal regulating system (the growth-hormone-IGF-I axis) are both markedly modified by training and these modifications are correlated with hemorheology. Nutritional disturbances affecting caloric and proteic intake, lipids, iron, zinc, etc. also modulate the hemorheologic effects of exercise. The overtraining syndrome represents a situation of unbalance between body's possibilities, nutrition, and work load, and is associated with metabolic, hormonal, immunologic and hemorheologic disturbances. The clinical relevance of these data is underlined by studies showing that exercise training in patients suffering from metabolic and/or cardiovascular disorders (such as the insulin resistance syndrome) results in a parallel improvement of metabolism, risk factors, blood rheology and fitness. Hemorheological measurements require to be studied, at least as sensitive markers of training, and possibly as "true" risk factors highly sensitive to exercise intensification. PMID- 12082248 TI - Does haemorheology explain the paradox of hypoxemia during exercise in elite athletes or thoroughbred horses? AB - Exercise-induced arterial hypoxemia (EIAH), i.e., a significant drop of O2 arterial partial pressure during sea level exercise, has been shown in both aerobically trained athletes and athletic animal species. The mechanisms potentially involved include ventilation/perfusion inequality and/or pulmonary diffusing capacity limitation. In thoroughbred horses, EIAH is going with pulmonary haemorrhage (EIPH). Stress failure of pulmonary capillaries leading to diffusion limitation has been proposed. Indeed, during intense exercise, the increased cardiac output and blood viscosity combine to rise capillary wall stress. Blood rheology may participate to the increase of VA/Q mismatch and capillary wall stress. High level of hematocrit (Hct) are known to alter blood flow distribution and rise shear stress in pulmonary capillaries. Any change in red blood cells (RBC) deformability may lead to aggregation at low shear rate, in post capillary veinules. There are contrasting data regarding the effects of blood rheology on EIPH in horses, however the large augmentation of hematocrit during exercise may cause vessel wall stress. In humans, greatest increase in hematocrit may participate to EIAH as well as RBC deformability. Today there is no consensus opinion and further studies of blood rheology in athletes is a field of interest. PMID- 12082249 TI - The microrheological behavior of young and old red blood cells in athletes. AB - Previous studies have shown a difference in rheological properties of young versus senescent RBCs. There are data that the athletes blood has more young RBCs than untrained people. Our research was a comparative study of the microrheological properties of young and old RBCs in athletes and in untrained people that was as control group. In athletes (men, n=24) and group of the control (men, n=20) the following parameters were measured: RBC aggregation (ARBC; Myrenne aggregometer) and deformability, RBC suspension and plasma viscosity as well as osmolarity, albumin, globulin and fibrinogen concentration, MCHC. Red cells were density (i.e., age) fractionated by the method of Murphy. After centrifugation the top 10% of the packed cell column (RBCtop, relatively young cells) and the bottom 10% (RBCbot, relatively old cells) were resuspended at 40.0+/-0.4% (in plasma) for aggregation, deformation and suspension viscosity measurements. It was found significant difference in aggregation and rigidity of the all RBC subpopulations between athletes and control group. The difference in aggregation was associated with reduced fibrinogen and increased ratio albumin/globulin in athletes. Besides, the correlation between aggregation RBCtop and RBCbot with fibrinogen was decreased in athletes. It was one of the cause of high fluidity of the RBCtop- and RBCbot suspensions and whole blood in athletes and more effective oxygen transport than in untrained people. PMID- 12082250 TI - Can white blood cell activation be one of the major factors that affect hemorheological parameters during and after exercise? PMID- 12082251 TI - Major alterations in body fluid status and blood rheology. AB - Since the dehydration causes a loss of body water, we studied the rheological properties of blood in the course of water deprivation. Subjects used in this study were 64 white male rats divided into 4 groups: control (n=19) and 3 experimental groups which underwent water deprivation for 3 days (n=15), 6 days (n=15) and 10 days (n=15). The results obtained indicate that under dehydration animals have higher blood and plasma viscosity and erythrocyte aggregation index than in the control group. After 3 days of dehydration these changes are due to the loss of intravascular water. The water deprivation for 10 days causes significant disturbances in blood composition as well as changes of red blood cell membrane properties whereas blood and plasma volume return to control values. PMID- 12082252 TI - Fluid shear stress induces the secretion of monocyte chemoattractant protein-1 in cultured human umbilical vein endothelial cells. AB - In this study we investigated the patterns of fluid shear stress on induction of monocyte chemoattractant protein-1 (MCP-1) secretion in cultured human umbilical vein endothelial cells (HUVECs). MCP-1 is a potent special chemoattractant, which recruits monocytes into the sub-endothelium. This process is one of the early events of atherosclerosis. We examined the pattern of fluid shear stress inducing the secretion of MCP-1 in cultured HUVECs from the view of biomechanics. In our experiments, HUVECs were subjected to controlled levels of shear stress (4, 10, 20 dyn/cm(2)) in a parallel plate flow chamber. MCP-1 in HUVECs of different periods was measured by an immunohistochemistry method and digital image analysis; MCP-1 in perfusion was measured by sandwich ELISA. The results demonstrated the increase of MCP-1 synthesis and secretion by shear stress was time- and force-dependent. The accumulated level of MCP-1 in HUVECs under lower shear stress (4 dyn/cm(2)) for 4-5 hrs was 3-fold compared with that for static cells. When the shear stress lasted to 6 hrs, the secretion of MCP-1 was reduced to normal levels and could not be increased even when the shear stress lasted for 12 hours. 10 dyn/cm(2) had less effect on the secretion of MCP-1 compared with 4 dyn/cm(2). This research provides data for understanding the mechanism of the contribution of hemodynamic forces to atherosclerosis. PMID- 12082253 TI - Insulin-like growth factor-binding protein 1 and blood rheology in athletes. AB - The GH-IGF axis has been recently suggested to modulate blood rheology in trained athletes, via GH effects on body water status and a possible action of IGF-I on erythrocyte deformability and aggregability. Another potential candidate for such a rheologic effect of the GH-IGF axis is insulin-like growth factor binding protein-1 (IGF-BP1) which is increased in trained people and correlated to fitness: IGF-BP1 is elevated in patients with polycythemia vera and stimulates erythroid burst formation in vitro. We investigated the statistical relationships between IGF-BP1 and blood rheology in athletes. 21 soccer players, age 24.5+/ 1.13 yr; body mass index 23.7+/-0.38 kg/m(2); VO2max 44.8+/-7 ml.min(-1).kg(-1)). The major statistical determinant of IGFBP1 (measured at rest after overnight fast) was age (r=0.752, p=0.00013) which was not correlated with rheological parameters. IGF BP1 was negatively correlated with blood viscosity eta (high shear rate r=-0.516, p=0.024) and positively correlated with the percentage of extracellular water in total body water (ECW/TBW) (r=0.488, p=0.039). The previously reported correlations between IGF-I and both eta (r=0.637, p=0.003) and red cell rigidity "Tk" (r=0.696, p=0.0137) were observed, but IGF-I and IGF BP1 were not correlated to each other (r=-0.176 ns) and their correlations with eta and Tk appeared to be independent when studied by multivariate analysis. Consistent with these correlations, subjects in the upper tertile of IGF-BP1 (>23.4 ng/ml) compared to those in the lower (<7.5 ng/ml) had a higher percentage of ECW/TBW (40.8+/-0.4 vs 38+/-0.8%, p=0.033), a lower eta (2.7+/-0.05 vs 2.97+/ 0.06 mPa.s, p=0.016), and a lower Tk (0.54+/-0.05 vs 0.63+/-0.01, p=0.027). Thus, beside GH and IGF-I, IGF-BP1, which is reported to act on erythroid progenitors, exhibits statistical relationships with blood fluidity and erythrocyte flexibility that may suggest a physiological role in improving blood rheology. PMID- 12082254 TI - Hormones, metabolism and body composition as major determinants of blood rheology: potential pathophysiological meaning. AB - The rheological properties of plasma and blood cells are markedly influenced by the surrounding milieu: physicochemical factors, metabolism and hormones. Acid/base status, osmolality, lipid status and plasma protein pattern are well known to exert a major influence. The oxidative stress induced by increased free radicals production decreases red cell deformability. Among circulating substances, the divalent cations magnesium and zinc improve red cell deformability probably via calcium antagonistic effects. Some metabolites like lactate or ketone bodies decrease red cell deformability, although the former has apparently the opposite effect in highly trained individuals. Endothelium-derived factors such as nitric oxide (NO) and several arachidonic acid derivatives modulate both RBC and white cell mechanics. Endothelium regulates also blood rheology via the release of PAI-1 which governs plasma fibrinogen levels. However, endothelium is not the only organ involved in the regulation of blood rheology: the kidney (by releasing erythropoietin which is a major "viscoregulatory" factor), the endocrine pancreas (via the action of insulin and glucagon on red cells), the adrenal gland (norepinephrine) and the endocrine heart (atrial natriuretic peptide) are also likely to exert important effects. Recently, increasing evidence is accumulating for a role of two other endocrine tissues in the regulation of blood rheology: the adipose tissue (free fatty acids, PAI-1, IL-6, leptin) and the pituitary gland (growth hormone-somatomedin axis, including the somatomedin carrier protein IGFBP1). These organs provide a link between body composition and hemorheology, since GH and somatomedins are major regulators of the body content in fat and water while the endocrine activity of fat mass is apparently proportional to its size. These mechanisms explain to some extent why many situations, either physiological (diet, exercise) or pathological (diabetes, uremia) are associated with marked changes in blood rheology that may in turn modify micro and macrocirculatory hemodynamics and the distribution of O(2) and fuels to tissues. PMID- 12082255 TI - Hemorheology and hypertension: not "chicken or egg" but two chickens from similar eggs. AB - A number of clinical studies have demonstrated significant positive correlation between the severity of arterial hypertension (AHT) and whole blood viscosity. Red blood cell aggregation has also been associated with AHT especially in the severe form of the disease. The main possible cause of increased red blood cell aggregation is fibrinogen which can be found in a significantly higher concentration in patients with AHT than in healthy controls. On the other hand, blood pressure reduction with angiotensin-converting-enzyme inhibitors, calcium channel-blocking agents, beta or alpha-receptor blocking drugs leads to a significant improvement of blood rheology. It can be presumed that abnormal hemorheology and AHT are not directly linked but they share the same inductive genetic and/or environmental factors like obesity, chronic mental stress, physical inactivity and cigarette smoking. Regarding this hypothesis, the appropriate question is not whether hemorheological factors are causes or results of AHT but what their common origins are. Further studies are needed to clarify this hypothetical link between hemorheology and AHT. PMID- 12082256 TI - The effect of dehydration on macro- and microrheological blood properties. AB - It is known that intensive physical activity causes a loss of water. Since physical capacity is highly dependent upon fluidity of blood, the rheological characteristics of which are largely determined by water content, the aim of our study was to estimate rheological properties of blood under dehydration. To model the state of dehydration, the water deprivation was performed on rats (n=49) for 3 and 10 days. We registered hematocrit, apparent plasma viscosity; apparent viscosity, rigidity index (Tk), structural parameter (k), and extent of aggregation ("M" and "M1" indices) were determined in both unfractionated and density separated RBC suspensions in autologous plasma at Ht=40. The Ht/viscosity ratio as index of oxygen supply to tissues was calculated. The results of our study indicated that water deprivation caused progressive reduction of blood and plasma fluidity. The increase of Ht (22%, p<0.001) and significant elevation of erythrocyte aggregation (205%, p<0.001) were observed. In spite of Ht increase, oxygen transport was decreased by 18% (p<0.05). RBC rigidity was increased under water deprivation (13.6%, p<0.05) as well as during aging. Significant correlation was found between indices of aggregation and Tk and k indices in the course of dehydration. Thus, our data indicated that dehydration caused impairment of blood and RBC rheological properties. PMID- 12082258 TI - Local RBC aggregation disturbing blood fluidity and causing stasis in microvessels. AB - Experiments in rat mesenterium were carried out under conditions when both pressure gradient in the chosen microvessels and their diameters were preserved constant. All details of the hemorheological events were directly visualized and documented by usage of appropriate microscopic video techniques. Intensified RBC aggregation locally produced in individual capillaries, immediately disturbs the normal blood flow structure inside their lumina and deranges the rheological properties of blood flow in the microvessels, which slows down till a full stop. The RBC aggregates gradually grow up due to addition of new cells, which become compressed and appear homogeneous. This usually interferes with restoration of blood flow in capillaries. Further the RBC aggregates can move slowly towards veins, while the flow accelerates immediately as soon as the aggregates reach the larger venules. PMID- 12082259 TI - Endothelial factors in the flux of atherogenic plasma proteins into artery walls. PMID- 12082257 TI - Rheological and biochemical examination of red blood cells after treatment with different pharmacological and toxic substances. AB - Rheological and enzymatic properties of red blood cells (RBC) were investigated in vitro after the treatment with vasoactive drug - buflomedil (bfl) and toxic substance - 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Studies with bfl included two different concentrations of the drug: 90 microg/ml of blood and 10 microg/ml of blood. The former concentration of the drug corresponds to the amount of bfl which is taken daily by a patient, the latter one is the highest peak of this drug in plasma. The dosage of dioxin was 32 microg/ml of blood and 32 ng/ml of blood. Only the smaller dosage of this compound appears in the environment but the higher one may occur in human organs because of its cumulation. Rheological properties of erythrocytes were examined using a laser diffractometer Rheodyne SSD (Myrenne). The deformability of RBC was expressed as an elongation index IE which was counted from the equation: EI=(L-W)/(L+W) where L is the length of cell and W is the width of cell. As far as the impact of bfl on RBC rheology is concerned studies were conducted in two different ways: (1) RBC were incubated with bfl directly, (2) RBC before incubation with bfl were treated with diamide to cause their rigidity. The action of bfl seems to be not efficient enough as data are not statistically significant in those two cases. Enzymatic properties of RBC were investigated using the methods of Beutler [7]. The activity of three enzymes was measured (acetylcholinesterase - Ache, dehydrogenase glucoso-6 phosphate - G-6-PD and gluthatione reductase - GR) for both bfl and TCDD-treated RBC. For TCDD-treated RBC additionally malonyldialdehyde (MDA) level was assessed. PMID- 12082260 TI - Effects of fluid shear stress on expression of proto-oncogenes c-fos and c-myc in cultured human umbilical vein endothelial cells. AB - The objective of our research was to reveal the effects of different shear stresses on the expression of proto-oncogenes c-fos and c-myc in cultured human umbilical vein endothelial cells (HUVEC). A parallel plate flow chamber was used to control the value and duration of shear stress (SS), and the expression of c fos and c-myc protein was measured by immunocytochemistry methods and image analysis software. Some important conclusions were drawn. In the stationary state, c-fos protein levels were very low. The SS's of 4 dyn/cm(2) and 10 dyn/cm(2) induced rapid increases of c-fos protein levels, especially the SS of 10 dyn/cm(2). The levels peaked at 1.0 h. Then, c-fos protein levels began to decrease, after 2.5 h, they declined to almost basal levels. In the stationary state, the c-myc protein levels were also very low slowly increasing after the onset of shear stress. The effects of 4 dyn/cm(2) and 10 dyn/cm(2) SS on c-myc protein expression levels had no difference and apparently were less than the effect on c-fos protein levels. The c-myc protein levels peaked at 1.5 h. Subsequently, they fell to basal levels at 2.5 h. Increased expression of these proto-oncogenes mediated by shear stress may have important effects on the regulation of critical cell activities, such as proliferation and differentiation. PMID- 12082261 TI - Effects of antihypertensive therapy on hemorheological profiles in female hypertensive patients with initially low or high whole blood viscosity. AB - This study was designed to examine changes of hemorheological parameters in essential arterial hypertension subjects following antihypertensive drug therapy. Eighty two female subjects were enrolled, and sub-divided into two groups based upon their high shear whole blood viscosity being lower (L) or higher (H) than normal controls. Equal numbers of L and H subjects were then treated for four weeks with one of four agents: angiotensin-converting enzyme inhibitor (ACE inhibitor, Spirapril - 6 mg/day); calcium antagonist (Isradipin - 5 mg/day); beta 1-blocker (Talinolol - 100 mg/day); diuretic (Indapamide - 1.5 mg/day). Both prior to and following drug treatment for six weeks, hemorheological measurements included plasma viscosity; high and low shear whole blood viscosity, hematocrit, fibrinogen and RBC aggregation. Treatment with each of the four drugs significantly (p<0.05) reduced blood pressure in both the L and H groups. However, the hemorheological effects of antihypertensive drug therapy differed markedly between groups: plasma and whole blood viscosity were significantly elevated in the L groups whereas these parameters were significantly decreased in the H groups. Fibrinogen levels and RBC aggregation decreased in both groups, whereas hematocrit was unaffected. These results thus suggest that the rheologic effects of antihypertensive drug therapy depend strongly on the initial, pre treatment status of the subject, and that for some subjects, such therapy can result in adverse hemorheological alterations. PMID- 12082262 TI - From biomechanics to mechanobiology. AB - Biomechanics can be defined as the application of mechanical concepts to the living world, and various fields of research have been developed such as the mechanics of movement, ergonomics, the mechanical properties of cells and tissues, and the relationship between physiology and applied forces. In this paper, the authors give, through several examples, an outline of these approaches and their potential biomedical applications, as in tissue remodelling, cell and tissue engineering and the development of biotissues. PMID- 12082263 TI - The functional environment of chondrocytes within cartilage subjected to compressive loading: a theoretical and experimental approach. AB - A non-invasive methodology (based on video microscopy, optimized digital image correlation and thin plate spline smoothing technique) has been developed to determine the intrinsic tissue stiffness (H(a)) and the intrinsic fixed charge density (c(0)(F)) distribution for hydrated soft tissues such as articular cartilage. Using this technique, the depth-dependent inhomogeneous parameters H(a)(z) and c(0)(F)(z) were determined for young bovine cartilage and incorporated into a triphasic mixture model. This model was then used to predict the mechanical and electrochemical events (stress, strain, fluid/osmotic pressure, and electrical potentials) inside the tissue specimen under a confined compression stress relaxation test. The integration of experimental measurements with theoretical analyses can help to understand the unique material behaviors of articular cartilage. Coupled with biological assays of cell-scale biosynthesis, there is also a great potential in the future to study chondrocyte mechanotransduction in situ with a new level of specificity. PMID- 12082264 TI - Proteoglycan deposition around chondrocytes in agarose culture: construction of a physical and biological interface for mechanotransduction in cartilage. AB - With a view towards the development of methods for cartilage tissue engineering, matrix deposition around individual chondrocytes was studied during de novo matrix synthesis in agarose suspension culture. At a range of times in culture from 2 days to 1 month (long enough for cartilage-like material properties to begin to emerge), pericellular distributions of proteoglycan and matrix protein deposition were measured by quantitative autoradiography, while matrix accumulation and cell volumes were estimated by stereological methods. Consistent with previous work, tissue-average rates of matrix synthesis generally decreased asymptotically with time in culture, as de novo matrix accumulated. Cell-scale analysis revealed that this evolution was accompanied by a transition from predominantly pericellular matrix (within a few microm from the cell membrane) deposition early in culture towards proteoglycan and protein deposition patterns more similar to those observed in cartilage explants at later times. This finding may suggest a differential recruitment of different proteoglycan metabolic pools as matrix assembly progresses. Cell volumes increased with time in culture, suggestive of alterations in volume regulatory processes associated with changes in the microphysical environment. Results emphasize a pattern of de novo matrix construction which proceeds outward from the pericellular matrix in a progressive fashion. These findings provide cell-scale insight into the mechanisms of assembly of matrix proteins and proteoglycans in de novo matrix, and may aid in the development of tissue engineering methods for cartilage repair. PMID- 12082265 TI - Electrical signals for chondrocytes in cartilage. AB - An important step toward understanding signal transduction mechanisms modulating cellular activities is the accurate predictions of the mechanical and electro chemical environment of the cells in well-defined experimental configurations. Although electro-kinetic phenomena in cartilage are well known, few studies have focused on the electric field inside the tissue. In this paper, we present some of our recent calculations of the electric field inside a layer of cartilage (with and without cells) in an open circuit one-dimensional (1D) stress relaxation experiment. The electric field inside the tissue derives from the streaming effects (streaming potential) and the diffusion effect (diffusion potential). Our results show that, for realistic cartilage material parameters, due to deformation-induced inhomogeneity of the fixed charge density, the two potentials compete against each other. For softer tissue, the diffusion potential may dominate over the streaming potential and vice versa for stiffer tissue. These results demonstrate that for proper interpretation of the mechano electrochemical signal transduction mechanisms, one must not ignore the diffusion potential. PMID- 12082266 TI - Measuring principles of frictional coefficients in cartilaginous tissues and its substitutes. AB - The frictional properties of cartilaginous tissues, such as the hydraulic permeability, the electro-osmotic permeability, the diffusion coefficients of various ions and solutes, and the electrical conductance, are vital data to characterise the extracellular environment in which chondrocytes reside. This paper analyses one-dimensional measurement principles of these coefficients. Particular attention is given to the deformation dependence of them and the highly deformable nature of the tissues. A suggested strategy is the combination of a diffusion experiment using radiotracer methods, an electro-osmotic flow experiment and an electro-osmotic pressure experiment at low electric current. PMID- 12082268 TI - Cell cytoskeleton and tensegrity. AB - The role of tensegrity architecture of the cytoskeleton in the mechanical behavior of living cells is examined by computational studies. Plane and spatial tensegrity models of the cytoskeleton are considered as well as their non tensegrity counterparts. Local buckling including deep postbuckling response of the compressed microtubules of the cytoskeleton is considered. The tensioned microfilaments cannot sustain compression. Large deformation of the whole model is accounted and fully nonlinear analysis is performed. It is shown that in the case of local buckling of the microtubules non-tensegrity models exhibit qualitatively the same linear stiffening as their tensegrity counterparts. This result raises the question of experimental validation of the local buckling of microtubules. If the microtubules of real cells are not straight, then tensegrity (in a narrow sense) is not a necessary attribute of the cytoskeleton architecture. If the microtubules are straight then tensegrity is more likely to be the cytoskeletal architecture. PMID- 12082267 TI - Influence of ion channels on the proliferation of human chondrocytes. AB - The goal of the study was to examine connections between ion channel activity and the proliferation of human chondrocytes. Chondrocytes were isolated form human osteoarthritic knee joint cartilage. In this study the concentration-dependent influence of the ion channel modulators tetraethylammonium (TEA), 4-aminopyridine (4-AP), 4',4' diisothiocyanato-stilbene-2,2'-disulfonic acid (DIDS), 4-acetamido 4'-isothiocyano-2,2'-disulfonic acid stilbene (SITS), verapamil (vp) and lidocaine (lido) on the membrane potential and the proliferation of human chondrocytes was investigated using flow cytometry and the measurement of (3)H thymidine incorporation as measure for the cell proliferation. The results show an effect of the used ion channel modulators causing a change of the membrane potential of human chondrocytes. The maximal measurable effects of the membrane potential were listed with 0.25 mmol/l verapamil (-18%) and 0.1 mmol/l lidocaine (+20%). When measuring DNA distribution, it became apparent that the human chondrocytes are diploid cells with a very low proliferation tendency. After 12 days culture duration, lidocaine and 4-AP cause an increase of the DNA synthesis rate being a limited effect. These results allow the conclusion of an influence of ion channel modulators on chondrocyte proliferation. To gain knowledge of the regulation of chondrocyte proliferation via ion channel modulators could serve the research of new osteoarthritis treatment concepts. PMID- 12082269 TI - Deformation properties of articular chondrocytes: a critique of three separate techniques. AB - This paper presents a series of techniques, which examine the deformation characteristics of bovine articular chondrocytes. The direct contact approach employs well established methodology, involving AFM and micropipette aspiration, to yield structural properties of local regions of isolated chondrocytes. The former technique yields a non-linear response with increased structural stiffness in a central location on a projected image of the chondrocyte. A simple viscoelastic model can be used with data from the micropipette aspiration technique to yield a mean value of Young's modulus, which is similar to that recently reported (Jones et al., 1999). An indirect approach is also described, involving the response of chondrocytes seeded within compressed agarose constructs. For 1% agarose constructs, the resulting cell strain, yields a gross cell modulus of 2.7 kPa. The study highlights the difficulties in establishing unique mechanical parameters, which reflect the deformation behaviour of articular chondrocytes. PMID- 12082270 TI - The influence of repair tissue maturation on the response to oscillatory compression in a cartilage defect repair model. AB - This study examined the effects of mechanical compression on engineered cartilage in a novel hybrid culture system. Cylindrical holes were cut in discs of bovine articular cartilage and filled with agarose gels containing chondrocytes. These constructs were compressed in radiolabeled medium under static or oscillatory unconfined compression. Oscillatory compression at 1 Hz significantly stimulated synthesis above static control levels. Control experiments indicate that oscillatory compression does not stimulate freshly cast gels (without annuli), but does so after several weeks. This may be because physiologic fluid flow levels do not occur until sufficient extracellular matrix has accumulated. Finite element models predict minimal fluid flow in the gel core, and minimal differences in flow patterns between free and constrained gels. However, the models predict fluid pressures in constrained gels to be substantially higher than those in free gels. Our results suggest that pressure variations may influence synthesis of engineered cartilage matrices, with implications for construct development and post-implantation survival. PMID- 12082271 TI - The role of fibril reinforcement in the mechanical behavior of cartilage. AB - Collagen fibril reinforcement was incorporated into a nonlinear poroelastic model for articular cartilage in unconfined compression. It was found that the radial fibrils play a predominant role in the transient mechanical behavior but a less important role in the equilibrium response of cartilage. The radial fibrils are in tension and can be highly stressed during compression, in contrast to low compressive stresses in all directions for the proteoglycan matrix after a small initial compression. The strain dependent fibril stiffening produces strong nonlinear transient response; the fibrils provide extra stiffness to balance a rising fluid pressure and to restrain stress increase in the proteoglycans. The fibril reinforcement, induced by the fluid pressure and flow, also accounts for a complex pattern of strain-magnitude and strain-rate dependence of cartilage stiffness. PMID- 12082272 TI - Differential responses of chondrocytes from normal and osteoarthritic human articular cartilage to mechanical stimulation. AB - Mechanical stimulation is critically important for the maintenance of normal articular cartilage integrity. Molecular events regulating responses of chondrocytes to mechanical forces are beginning to be defined. Chondrocytes from normal human knee joint articular cartilage show increased levels of aggrecan mRNA following 0.33 Hz mechanical stimulation whilst at the same time relative levels of MMP3 mRNA are decreased. This anabolic response, associated with membrane hyperpolarisation, is activated via an integrin-dependent interleukin (IL)-4 autocrine/paracrine loop. Work in our laboratory suggests that this chondroprotective response may be aberrant in osteoarthritis (OA). Chondrocytes from OA cartilage show no changes in aggrecan or MMP3 mRNA following 0.33 Hz mechanical stimulation. alpha5beta1 integrin is the mechanoreceptor in both normal and OA chondrocytes but downstream signalling pathways differ. OA chondrocytes show membrane depolarisation following 0.33 Hz mechanical stimulation consequent to activation of an IL1beta autocrine/paracrine loop. IL4 signalling in OA chondrocytes is preferentially through the type I (IL4alpha/cgamma) receptor rather than via the type II (IL4alpha/IL13R) receptor. Altered mechanotransduction and signalling in OA may contribute to changes in chondrocyte behaviour leading to increased cartilage breakdown and disease progression. PMID- 12082273 TI - High pressure effects on cellular expression profile and mRNA stability. A cDNA array analysis. AB - Hydrostatic pressure has a profound effect on cartilage tissue and chondrocyte metabolism. Depending on the type and magnitude of pressure various responses can occur in the cells. The mechanisms of mechanotransduction at cellular level and the events leading to specific changes in gene expression are still poorly understood. We have previously shown that induction of stress response in immortalized chondrocytes exposed to high static hydrostatic pressure increases the stability of heat shock protein 70 mRNA. In this study, our aim was to examine the effect of high pressure on gene expression profile and to study whether stabilization of mRNA molecules is a general phenomenon under this condition. For this purpose a cDNA array analysis was used to compare mRNA expression profile in pressurized vs. non-pressurized human chondrosarcoma cells (HCS 2/8). mRNA stability was analyzed using actinomycin-treated and nontreated samples collected after pressure treatment. A number of immediate-early genes, and genes regulating cell cycle and growth were up-regulated due to high pressure. Decrease in osteonectin, fibronectin, and collagen types VI and XVI mRNAs was observed. Also bikunin, cdc37 homologue and Tiam1, genes linked with hyaluronan metabolism, were down-regulated. In general, stability of down regulated mRNA species appeared to increase. However, no increase in mRNA above control level due to stabilization was noticed in the genes available in the array. On the other hand, mRNAs of certain immediate-early genes, like c-jun, jun B and c-myc, became destabilized under pressure treatment. Increased accumulation of mRNA on account of stabilization under high pressure conditions seems to be a tightly regulated, specific phenomenon. PMID- 12082274 TI - Integrins and cell signaling in chondrocytes. AB - Integrins are adhesion receptor heterodimers that transmit information from the extracellular matrix (ECM) to the cell through activation of cell signaling pathways. Chondrocytes express several members of the integrin family including alpha5beta1 which is the primary chondrocyte receptor for fibronectin. Cell signaling mediated through integrins regulates several chondrocyte functions including differentiation, matrix remodeling, responses to mechanical stimulation and cell survival. Integrin-mediated activation of members of the mitogen activated protein kinase family likely plays a key role in transmitting signals regulating chondrocyte gene expression. Upstream mediators of mitogen-activated protein kinase (MAP kinase) activation include focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (pyk2) which are both expressed by chondrocytes. A better understanding of chondrocyte integrin signaling is needed to define the mechanisms by which the ECM regulates chondrocyte function. PMID- 12082275 TI - The effects of pressure on chondrocyte tumour necrosis factor receptor expression. AB - This work was performed to determine whether one aspect of load, pressure, could alter tumour necrosis factor (TNF) receptor type I (RI) expression on chondrocytes. Encapsulated tsT/AC62, osteoarthritic (OA) or non-arthritic (NA) chondrocytes were centrifuged at speeds representing 5 or 20 MPa, incubated for specific periods, released from alginate and TNFRI and II (TNFRII) expression determined by flow cytometry. Significant (p<0.05, n=4) changes in tsT/AC62 chondrocyte TNFRI expression were apparent 24 hours after application of 20 MPa. Five or 20 MPa increased OA chondrocyte TNFRI expression; chondrocytes from some OA patients were markedly sensitive to 20 MPa. NA chondrocyte TNFRI expression usually decreased in response to 5 and 20 MPa. Significant pressure-induced differences in TNFRI expression between NA and OA groups were apparent at 5, but not 20 MPa. Pressure did not significantly alter TNRFII expression on tsT/AC62, NA or OA chondrocytes. These results suggest a mechanism whereby sensitivity of chondrocytes to the effects of TNFalpha may be increased, in susceptible individuals, in regions of the joint that experience peak loading. PMID- 12082276 TI - The effect of mechanical stress on cartilage energy metabolism. AB - Cartilage is routinely subjected to varying mechanical stresses which are known to affect matrix turnover by a variety of pathways. Here we show that mechanical loads which suppress sulphate incorporation or protein synthesis by articular chondrocytes, also inhibit rates of oxygen uptake and of lactate production. Although the mechanisms have not been definitively identified, it has been shown that high hydrostatic pressures reduce the activity of the glucose transporter GLUT. Furthermore, fluid expression consequent on static loading changes intracellular pH and ionic strength; intracellular changes which would reduce the activity of glycolytic enzymes. Both pathways would thus lead to a fall in rates of glycolysis and a reduction in intracellular ATP, and - since ATP concentrations directly affect sulphation of proteoglycans - a rapid fall in sulphate incorporation. Our results suggest that load-induced changes in matrix synthesis in cartilage can occur by means other than changes in gene expression. PMID- 12082277 TI - Human chondrocyte senescence and osteoarthritis. AB - Although osteoarthritis (OA) is not an inevitable consequence of aging, a strong association exists between age and increasing incidence of OA. We hypothesized that this association is due to in vivo articular cartilage chondrocyte senescence which causes an age-related decline in the ability of the cells to maintain articular cartilage, that is, increasing age increases the risk of OA because chondrocytes lose their ability to replace their extracellular matrix. To test this hypothesis, we measured senescence markers in human articular cartilage chondrocytes from 27 donors ranging in age from one to 87 years. The markers included expression of the senescence-associated enzyme beta-galactosidase, mitotic activity measured by 3H-thymidine incorporation, and telomere length. beta-galactosidase expression increased with age (r=0.84, p=0.0001) while mitotic activity and mean telomere length declined (r=-0.774, p=0.001 and r=-0.71, p=0.0004, respectively). Decreasing telomere length was strongly correlated with increasing expression of beta-galactosidase and decreasing mitotic activity. These findings help explain the previously reported age related declines in chondrocyte synthetic activity and responsiveness to anabolic growth factors and indicate that in vivo articular cartilage chondrocyte senescence is responsible, at least in part, for the age related increased incidence of OA. The data also imply that people vary in their risk of developing OA because of differences in onset of chondrocyte senescence; and, the success of chondrocyte transplantation procedures performed to restore damaged articular surfaces in older patients could be limited by the inability of older chondrocytes to form new cartilage. New efforts to prevent the development or progression of OA might include strategies that delay the onset of chondrocyte senescence or replace senescent cells. PMID- 12082278 TI - The functional expression of connexin 43 in articular chondrocytes is increased by interleukin 1beta: evidence for a Ca2+-dependent mechanism. AB - Cell-to-cell interactions and gap junctions-dependent communication are crucially involved in chondrogenic differentiation, while in adult articular cartilage direct intercellular communication occurs mainly among chondrocytes facing the outer cartilage layer. Chondrocytes extracted from adult articular cartilage and grown in primary culture express connexin 43 and form functional gap junctions capable of sustaining the propagation of intercellular Ca2+ waves. Degradation of articular cartilage is a characteristic feature of arthritic diseases and is associated to increased levels of interleukin-1 (IL-1) in the synovial fluid. We have examined the effects of IL-1 on gap junctional communication in cultured rabbit articular chondrocytes. Incubation with IL-1 potentiated the transmission of intercellular Ca2+ waves and the intercellular transfer of Lucifer yellow. The stimulatory effect was accompanied by a dose-dependent increase in the expression of connexin 43 and by an enhanced connexin 43 immunostaining at sites of cell-to cell contact. IL-1 stimulation induced a dose-dependent increase of cytosolic Ca2+ and activates protein tyrosine phosphorylation. IL-1-dependent up-regulation of connexin 43 could be prevented by intracellular Ca2+ chelation, but not by inhibitors of protein tyrosine kinases, suggesting a crucial role of cytosolic Ca2+ in regulating the expression of connexin 43. IL-1 is one of the most potent cytokines that promotes cartilage catabolism: its modulation of intercellular communication represents a novel mechanism by which proinflammatory mediators regulate the activity of cartilage cells. PMID- 12082279 TI - Ultrasonic characterization of articular cartilage. AB - Osteoarthrosis is the most important joint disease that threatens health of the musculoskeletal system of elderly people. Today, there is a need for sensitive, quantitative diagnostic methods for successful and early diagnosis of the disorder. In the present study, we aimed at evaluating the applicability of ultrasound for quantitative assessment of cartilage structure and properties. Bovine articular cartilage was investigated both in vitro and in situ using high frequency ultrasound. Cartilage samples were also tested mechanically in vitro to reveal relationships between acoustic and mechanical parameters of the tissue. The collagen organization and proteoglycan content of cartilage samples were mapped, using quantitative polarized light microscopy and digital densitometry, respectively, to reveal their effect on the acoustic properties of tissue. The high frequency pulse-echo ultrasound (20-30 MHz) technique proved to be sensitive in detecting the degeneration of the superficial collagen-rich cartilage zone. In addition, ultrasound was found to be a potential tool for measuring cartilage thickness. When the results from biomechanical indentation measurements and ultrasound measurements of normal and enzymatically degraded articular cartilage were combined, collagen or proteoglycan degradation in the tissue could be sensitively and specifically differentiated from each other. To conclude, high frequency ultrasound is a useful tool for evaluation of the quality of superficial articular cartilage as well as for the measurement of cartilage thickness. Therefore, ultrasound appears to be a valuable supplement to the mechanical measurements of articular cartilage stiffness. PMID- 12082280 TI - COX-1, COX-2 and articular joint disease: a role for chondroprotective agents. AB - It is widely accepted that whilst exhibiting clinically useful anti-inflammatory and analgesic activity, the application of non-steroidal anti-inflammatory drugs (NSAIDs) does not affect the underlying pathogenesis of articular diseases such as rheumatoid arthritis. The demonstration of a role for COX-2 in the resolution of inflammation may partly underly the lack of disease modifying activity seen with NSAIDs in long term use in these inflammatory joint diseases. This has led to the suggestion that the anti-arthritic efficacy of these agents may be improved by altering prescribing practice such that they are not given during periods of disease remission, which may be difficult to achieve in the clinic. Alternatively, they may benefit from concomitant administration of chondroprotective agents, such as diacetylrhein, which may protect against the deleterious effects of traditional NSAIDs on cartilage degradation and, further, inhibit additional pathways such as cytokine elaboration which are important in joint destruction. PMID- 12082281 TI - The influence of exercise on the composition of developing equine joints. AB - An overview is given of the direct and long-term effects of exercise on the biochemical characteristics of cartilage and subchondral bone, and on the metabolic activity of chondrocytes in the juvenile horse. In the experimental setup 43 foals were reared until weaning at 5 months of age under similar conditions, except for the type and amount of exercise. Fifteen foals remained at pasture (Pasture group and also control group), 14 foals were kept in box stalls (Box group), and 14 foals were kept in the same box stalls but were subjected daily to an increasing number of gallop sprints (Training group). After weaning 8 foals from each group were euthanised. All remaining 19 animals were housed together in a loose box with access to a small paddock to study a possible reversibility of exercise-induced effects. Post mortem subchondral bone and cartilage samples were collected and analysed for bone morphogenic enzymes, matrix composition, chondrocyte metabolic activity, and bone mineral density. It resulted that lack of exercise leads to a retardation of the normal development of the joint. This is largely compensated for when afterwards a more normal exercise regimen is followed. Most parameters in the Training group approximated those of the pastured foals at age 5 months. However, at age 11 months there were indications for a reduced performance of the investigated tissues in this group. It is concluded that regular, sub-maximal loading, as occurred in the Pasture group, seems best for an optimal development of the musculoskeletal tissues. The combination of short bouts of heavy exercise superimposed on a basic box rest regimen appears to have adverse effects on long-term viability of the tissues and may hence lead to an impaired resistance to injury. PMID- 12082282 TI - Tensile fatigue behaviour of articular cartilage. AB - Although fatigue has been implicated in cartilage failure there are only two studies by the same author, and in both of which cartilage was tested in the direction parallel to the collagen orientation in the surface layer. In the present work articular cartilage was tested also along the perpendicular direction, being the direction in which cartilage possesses lower tensile strength. Specimens were tested under cyclic tensile load. Number of cycles at failure was recorded as well as elongation of the specimen. To date 72 specimens have been tested all from one knee joint. The number of cycles to failure ranged between two and 1.5 million. The surface and deep layers have better fatigue properties whether tested in the parallel or the perpendicular direction, while the middle layer was far weaker. Better fatigue behaviour was observed with specimens tested in parallel than in perpendicular direction to the fibres. PMID- 12082284 TI - Growth factors in cartilage tissue engineering. AB - Tissue engineering of cartilage consists of two steps. Firstly, the cells from a small biopsy of patient's own tissue have to be multiplied. During this multiplication process they lose their cartilage phenotype. In the second step, these cells have to be stimulated to re-express their cartilage phenotype and produce cartilage matrix. Growth factors can be used to improve cell multiplication, redifferentiation and production of matrix. The choice of growth factors should be made for each phase of the tissue engineering process separately, taking into account cell phenotype and the presence of extracellular matrix. This paper demonstrates some examples of the use of growth factors to increase the amount, the quality and the assembly of the matrix components produced for cartilage tissue engineering. In addition it shows that the "culture history" (e.g., addition of growth factors during cell multiplication or preculture period in a 3-dimensional environment) of the cells influences the effect of growth factor addition. The data demonstrate the potency as well as the limitations of the use of growth factors in cartilage tissue engineering. PMID- 12082283 TI - Nitric oxide (NO) and cartilage metabolism: NO effects are modulated by superoxide in response to IL-1. AB - Nitric oxide (NO) is thought to mediate most effects of interleukin-1 (IL-1) on cartilage. In vitro evidence includes the decreased synthesis of extracellular matrix components, the abnormal cell renewal, the decreased production of IL-1 receptor antagonist, the induction of apoptosis and the enhanced sensitivity of chondrocytes to oxidative stress. Studies in NOS2(-/-) mice or administration of NO synthase inhibitors in animal models of joint disorders have confirmed its potent pathophysiological role in cartilage. Using L-NMMA (1 mM), as a NO synthase inhibitor, and CuDips (10 microM), as a SOD mimetic, we provide evidence that the inhibitory potency of IL-1beta on proteoglycan synthesis and its stimulating effect on COX-2 activity depend both on NO and O2-* production. Peroxynitrite formation is further demonstrated by the occurrence of 3 nitrotyrosines in chondrocytes stimulated in vitro with 2.5 ng/ml IL-1 and in femoral condyles of rats injected locally with 1 microg IL-1. Preliminary data suggest that such contribution of reactive oxygen species is not shared in common by IL-17, another NO-producing cytokine. We conclude that superoxide is a key modulator of NO-mediated effects in chondrocyte stimulated with IL-1 and that a combined therapy with NO synthase inhibitors and antioxidants may be promising for a full cartilage protection. PMID- 12082285 TI - Animal models of osteoarthritis. AB - Animal models have proved to be of considerable importance in elucidating mechanisms underlying joint damage in osteoarthritis (OA) and providing proof of concept in the development of pharmacologic and biologic agents that may modify structural damage in the OA joint. The utility of animal models in predicting the response to an intervention with a drug or biologic agent in humans, however, can be established only after evidence is obtained of a positive effect of the agent in humans. To date, no agent has been shown unequivocally to have such an effect, although diacerhein and glucosamine have recently been reported to lower the rate of loss of articular cartilage in patients with hip OA and knee OA, respectively, based on measurements of the rate of joint space narrowing in plain radiographs. Furthermore, the predominant manifestation of OA - and the feature that leads people with radiographic changes of the disease to decide to seek medical attention and contributes to the enormous medicoeconomic and socioeconomic burden imposed by the disease - is joint pain. Notably, none of the animal models of OA is a good indicator of the analgesic effects of pharmacologic agents. Indeed, it should not be assumed a priori that reduction in the rate of progression of joint damage in OA will be associated with a reduction in joint pain. PMID- 12082286 TI - The role of cytokines in osteoarthritis pathophysiology. AB - Morphological changes observed in OA include cartilage erosion as well as a variable degree of synovial inflammation. Current research attributes these changes to a complex network of biochemical factors, including proteolytic enzymes, that lead to a breakdown of the cartilage macromolecules. Cytokines such as IL-1 and TNF-alpha produced by activated synoviocytes, mononuclear cells or by articular cartilage itself significantly up-regulate metalloproteinases (MMP) gene expression. Cytokines also blunt chondrocyte compensatory synthesis pathways required to restore the integrity of the degraded extrecellular matrix (ECM). Moreover, in OA synovium, a relative deficit in the production of natural antagonists of the IL-1 receptor (IL-1Ra) has been demonstrated, and could possibly be related to an excess production of nitric oxide in OA tissues. This, coupled with an upregulation in the receptor level, has been shown to be an additional enhancer of the catabolic effect of IL-1 in this disease.IL-1 and TNF alpha significantly up-regulate MMP-3 steady-state mRNA derived from human synovium and chondrocytes. The neutralization of IL-1 and/or TNF-alpha up regulation of MMP gene expression appears to be a logical development in the potential medical therapy of OA. Indeed, recombinant IL-1receptor antagonists (ILRa) and soluble IL-1 receptor proteins have been tested in both animal models of OA for modification of OA progression. Soluble IL-1Ra suppressed MMP-3 transcription in the rabbit synovial cell line HIG-82. Experimental evidence showing that neutralizing TNF-alpha suppressed cartilage degradation in arthritis also support such strategy. The important role of TNF-alpha in OA may emerge from the fact that human articular chondrocytes from OA cartilage expressed a significantly higher number of the p55 TNF-alpha receptor which could make OA cartilage particularly susceptible to TNF-alpha degradative stimuli. In addition, OA cartilage produces more TNF-alpha and TNF anglealpha convertase enzyme (TACE) mRNA than normal cartilage. By analogy, an inhibitor to the p55 TNF-alpha receptor may also provide a mechanism for abolishing TNF-alpha-induced degradation of cartilage ECM by MMPs. Since TACE is the regulator of TNF-alpha activity, limiting the activity of TACE might also prove efficacious in OA. IL-1 and TNF-alpha inhibition of chondrocyte compensatory biosynthesis pathways which further compromise cartilage repair must also be dealt with, perhaps by employing stimulatory agents such as transforming growth factor-beta or insulin-like growth factor-I. Certain cytokines have antiinflammatory properties. Three such cytokines - IL-4, IL-10, and IL-13 - have been identified as able to modulate various inflammatory processes. Their antiinflammatory potential, however, appears to depend greatly on the target cell. Interleukin-4 (IL-4) has been tested in vitro in OA tissue and has been shown to suppress the synthesis of both TNF-alpha and IL-1beta in the same manner as low-dose dexamethasone. Naturally occurring antiinflammatory cytokines such as IL-10 inhibit the synthesis of IL-1 and TNF-alpha and can be potential targets for therapy in OA. Augmenting inhibitor production in situ by gene therapy or supplementing it by injecting the recombinant protein is an attractive therapeutic target, although an in vivo assay in OA is not available, and its applicability has yet to be proven. Similarly, IL-13 significantly inhibits lipopolysaccharide (LPS)-induced TNF alpha production by mononuclear cells from peripheral blood, but not in cells from inflamed synovial fluid. IL-13 has important biological activities: inhibition of the production of a wide range of proinflammatory cytokines in monocytes/macrophages, B cells, natural killer cells and endothelial cells, while increasing IL-1Ra production. In OA synovial membranes treated with LPS, IL-13 inhibited the synthesis of IL-1beta, TNF-alpha and stromelysin, while increasing IL-1Ra production.In summary, modulation of cytokines that control MMP gene up regulation would appear to be fertile targets for drug development in the treatment of OA. Several studies illustrate the potential importance of modulating IL-1 activity as a means to reduce the progression of the structural changes in OA. In the experimental dog and rabbit models of OA, we have demonstrated that in vivo intraarticular injections of the IL-Ra gene can prevent the progression of structural changes in OA. Future directions in the research and treatment of osteoarthritis (OA) will be based on the emerging picture of pathophysiological events that modulate the initiation and progression of OA. PMID- 12082287 TI - Differential gene expression analysis in a rabbit model of osteoarthritis induced by anterior cruciate ligament (ACL) section. AB - Osteoarthritis (OA) is the most common of all joint diseases to affect mankind and is characterized by the degradation of articular cartilage. The low availability of normal and pathologic human cartilage and the inability to study the early stages of the disease in humans has led to the development of numerous animal models of OA. The aim of our study was to establish gene expression profiles during the progression of a rabbit model of OA induced by anterior cruciate ligament (ACL) section. Semiquantitative RT-PCR was used to follow expression of several relevant molecules (type II and X collagens, aggrecan, osteonectin, betaig-h3, BiP, TIMP-1, MMP-1, -3, -13, aggrecanase-1, -2) during development of OA in articular cartilage. In parallel, we monitored the activities of collagenase, caseinase, phospholipase A2 and glycosyltransferases (xylosyl-, galactosyl-, glucuronyl- and N-acetyl-galactosaminyl-transferase). Novel cDNA clones for rabbit type X collagen, aggrecanase-1 and -2, osteonectin and BiP were constructed to obtain species-specific primers. Ours result show that MMP-13 (collagenase-3) gene expression increased dramatically early after ACL surgery and remained high thereafter. An increase in MMP-1 (collagenase-1) and MMP-3 expression was also noted with an absence of variation for TIMP-1 expression. In addition, the global MMPs activities paralleled the MMP gene expression. These data together characterize at the molecular level the evolution of OA in this rabbit model. Furthermore, we have undertaken a search for identifying differentially expressed genes in normal and OA cartilage in this model, by differential display RT-PCR. We present here preliminary results with the determination of the best technical conditions to obtain reproducible electrophoresis patterns of differential display RT-PCR. PMID- 12082288 TI - Bioreactor studies of native and tissue engineered cartilage. AB - Functional tissue engineering of cartilage involves the use of bioreactors designed to provide a controlled in vitro environment that embodies some of the biochemical and physical signals known to regulate chondrogenesis. Hydrodynamic conditions can affect in vitro tissue formation in at least two ways: by direct effects of hydrodynamic forces on cell morphology and function, and by indirect flow-induced changes in mass transfer of nutrients and metabolites. In the present work, we discuss the effects of three different in vitro environments: static flasks (tissues fixed in place, static medium), mixed flasks (tissues fixed in place, unidirectional turbulent flow) and rotating bioreactors (tissues dynamically suspended in laminar flow) on engineered cartilage constructs and native cartilage explants. As compared to static and mixed flasks, dynamic laminar flow in rotating bioreactors resulted in the most rapid tissue growth and the highest final fractions of glycosaminoglycans and total collagen in both tissues. Mechanical properties (equilibrium modulus, dynamic stiffness, hydraulic permeability) of engineered constructs and explanted cartilage correlated with the wet weight fractions of glycosaminoglycans and collagen. Current research needs in the area of cartilage tissue engineering include the utilization of additional physiologically relevant regulatory signals, and the development of predictive mathematical models that enable optimization of the conditions and duration of tissue culture. PMID- 12082289 TI - The biochemical content of articular cartilage: an original MRI approach. AB - The MR aspect of articular cartilage, that reflects the interactions between protons and macromolecular constituents, is affected by the intrinsic tissue structure (water content, the content of matrix constituents, collagen network organization), imager characteristics, and acquisition parameters. On the T1 weighted sequences, the bovine articular cartilage appears as an homogeneous tissue in high signal intensity, whatever the age of animals considered, whereas on the T2-weighted sequences, the articular bovine cartilage presents variations of its imaging pattern (laminar appearance) well correlated to the variations of its histological and biochemical structure. The T2 relaxation time measurement (T2 mapping), which reflects quantitatively the signal intensity variations observed on T2 weighted sequences, is a way to evaluate more precisely the modifications of cartilage structure during the aging and maturation processes (rat's study). This technique so far confined to experimental micro-imagers is now developed on clinical imagers. Consequently, it may permit to depict the early stages of osteoarthritic disease (OA) or to evaluate the chondroprotective effect of drugs. PMID- 12082290 TI - In vitro study of intracellular IL-1beta production and beta1 integrins expression in stimulated chondrocytes--effect of rhein. AB - The purpose of the present study was to investigate the intracellular IL-1beta production and beta1 integrins (alpha4/beta1 and alpha5/beta1) expression on chondrocytes. Chondroytes monolayer (human chondrosarcoma cell line HEM-C55) were incubated for 12, 24 and 48 hours in the presence of Tumor Necrosis Factor-alpha (TNF-alpha, Sigma, France) or recombinant human IL-1alpha (rh-IL1alpha, Becton Dickinson, France). After direct immunolabelling, cells were either analyzed on FACScan flow cytometer (Becton Dickinson, France), or observed under an epi fluorescence inverted microscope equipped with the CellScan EPR optical scanning acquisition system (IPLab-Scanalytics, USA). We found that the IL-1beta mean fluorescence intensity in flow cytometry and in 3D microscopy was increased in the presence of TNF-alpha or rh-IL-1alpha, and alpha4/beta1 or alpha5/beta1 expression was higher on stimulated cells than on control cells. On the other hand, we have evaluated the in vitro effects of rhein (10(-5) M, Negma, France), an active metabolite of diacerein, on the intracellular IL-1beta and beta1 integrins expressed by stimulated or no-stimulated chondrocytes. The results indicated that rhein leads to a reduction of IL-1beta synthesis whereas a weak decrease of beta1 integrins receptors expression is observed. From this study, it seems that rhein partially reduce cytokine-induced intracellular IL-1beta production, and it has a weak action on alpha4/beta1 or alpha5/beta1 receptors. PMID- 12082292 TI - Ovarian carcinoma and serous effusions. Changing views regarding tumor progression and review of current literature. AB - Carcinoma of the ovary is the leading cause of death from gynecological cancer in western countries. Ovarian carcinoma is commonly associated with the accumulation of fluid containing malignant cells in the peritoneal, and not infrequently in the pleural cavity. The differentiation of these cells from reactive mesothelial cells is at times difficult. In addition, tumor progression in ovarian carcinoma and the biological characteristics of carcinoma cells in effusions compared to their counterparts in solid tumors are poorly understood. This review details the current knowledge regarding diagnostic and biologic aspects of effusion cytology, with emphasis on ovarian carcinoma. Results from our first studies of effusions are subsequently presented. These attempt to address several issues. First, to improve the diagnostic ability to detect cancer cells in effusions using antibodies designed for the differentiation of epithelial cells from mesothelial cells. Secondly, to study genotypic and phenotypic differences between ovarian carcinoma cells in effusions, solid primary tumors and metastatic lesions, as well as to compare malignant cells in peritoneal and pleural effusions. These studies of carbohydrate antigens, E-cadherin complex and matrix metalloproteinases (MMP) attempted to evaluate whether ovarian carcinoma cells in effusions possess true metastatic properties, or are similar to the cells in primary tumors, thereby merely representing the result of a shedding process. Finally, the prognostic role of these molecules was studied in solid tumors from a patient cohort consisting of long- and short-term survivors, followed for up to 20 years. Figure 1 on http://www.esacp.org/acp/2001/23-3,4/davidson.htm. PMID- 12082291 TI - Prognostic significance of DNA cytometry in carcinoma of the uterine cervix FIGO stage IB and II. AB - OBJECTIVE: To assess the prognostic value of DNA-image cytometry in cervical carcinoma of the uterus and its relation to other established prognostic factors. STUDY DESIGN: The study included 116 cases of cervical carcinoma FIGO stages IB and II which were treated with radical abdominal hysterectomy. The median follow up was 55 months (range 1-162 months). DNA image cytometry was performed on cytologic specimens prepared by enzymatic cell separation from formalin-fixed, paraffin-embedded tissues. DNA stemline ploidy, DNA stemline aneuploidy, 5c exceeding rate, 9c exceeding rate, 2c deviation index, and DNA malignancy grade were computed. DNA-variables as well as various clinical and histological variables were related to survival rates. RESULTS: In multivariate statistical analysis DNA stemline ploidy using 2.2c as a cut-off value and FIGO stage showed to be statistically significant available presurgery predictors of survival, whereas the postsurgical parameters lymphonodal status, tumor size and parametrial involvement were significantly correlated with survival. The synopsis of all parameters in a multivariate Cox model indicated that - with declining relevance - the number of positive pelvic lymph nodes, DNA stemline ploidy using a cut-off level at a modal value of 2.2c, largest pelvic lymph node, 5c exceeding rate, and ratio of carcinoma area to cervix area, were of predictive value for survival. CONCLUSIONS: Our results suggest that prognostic information deducted from classical staging parameters is successfully complemented by DNA image cytometry which can be applied pretherapeutically. PMID- 12082293 TI - Application of neural networks to the classification of pancreatic intraductal proliferative lesions. AB - The aim of the study was to test applycability of neural networks to classification of pancreatic intraductal proliferative lesions basing on nuclear features, especially chromatin texture. Material for the study was obtained from patients operated on for pancreatic cancer, chronic pancreatitis and other tumours requiring pancreatic resection. Intraductal lesions were classified as low and high grade as previously described. The image analysis system consisted of a microscope, CCD camera combined with a PC and AnalySIS v. 2.11 software. The following texture characteristics were measured: variance of grey levels, features extracted from the grey levels correlation matrix and mean values, variance and standard deviation of the energy obtained from Laws matrices. Furthermore we used moments derived invariants and basic geometric data such as surface area, the minimum and maximum diameter and shape factor. The sets of data were randomly divided into training and testing groups. The training of the network using the back-propagation algorithm, and the final classification of data was carried out with a neural network simulator SNNS v. 4.1. We studied the efficacy of networks containing from one to three hidden layers. Using the best network, containing three hidden layers, the rate of correct classification of nuclei was 73%, and the rate of misdiagnosis was 3%; in 24% the network response was ambiguous. The present findings may serve as a starting point in search for methods facilitating early diagnosis of ductal pancreatic carcinoma. PMID- 12082294 TI - RNA relocation and persistence of nucleolus-like bodies at mitosis in benzo[a]pyrene-transformed human breast epithelial cells after microcell-mediated transfer of chromosomes 11 and 17. AB - RNA relocation and the incidence of nucleolus-like bodies accumulated during mitosis were studied cytochemically in benzo[a]pyrene (BP)-transformed human breast epithelial MCF-10F cells after microcell-mediated transfer of normal chromosomes 11 and 17. The changes resulting from the transfer of these two chromosomes in tumorigenic MCF-10F cells (BP1-E cell line) were examined, since alterations in these chromosomes are involved in the expression of the transformed and tumorigenic phenotypes in the MCF-10F cell series. In addition, the frequency of nucleolus-like bodies decreases drastically with transformation and tumorigenicity in MCF-10F cells, thus being conceivable that it would be affected in presence of normal chromosomes 11 or 17. The pattern of RNA relocation associated with the mitotic spindle did not vary in the cell lines analyzed. The introduction of chromosome 17 in BP1-E cells either decreased or did not affect the frequency of persistent nucleolus-like bodies. In contrast, in cells which received a normal chromosome 11, the frequency of nucleolus-like bodies was closer to that of non-transformed MCF-10F cells. These results suggest that a normal chromosome 11 but not chromosome 17 contributes to the maintenance of an RNA surplus which accumulates in nucleolus-like bodies during cell division of the human breast epithelial cells, at least in vitro. Some loci which were retained in the BP1-E cells which received a normal chromosome 11 are probably involved with the control of RNA transcript production. Figure 1 on http://www.esacp.org/acp/2001/23-3,4/mello.htm PMID- 12082295 TI - DNA ploidy and S-phase fraction in carcinoma of the gallbladder related to histopathology, number of gallstones and survival. AB - Gallstones are a risk factor for the development of gallbladder cancer. We studied DNA ploidy and cell cycle composition by flow cytometry in archival specimens from 52 gall bladder carcinomas in relation to histopathological grade, tumour stage, gallstone number and survival. 69% of the gallbladder carcinomas showed aneuploidy. All tumours with single stones (N=11) were aneuploid while only 61% of tumours with multiple stones (N=41) were aneuploid (p=0.002). DNA aneuploidy was related to increase in T-category (p=0.01), grade (p=0.02), and nuclear pleomorphism (p=0.0005). The distribution of DNA ploidy shifted from tetraploid in low stage towards triploid positions in high stage tumours (p=0.02) combined with higher S-phase values in triploid tumours (p=0.05). S-phase fraction increased during development from normal tissue to dysplasia, cancer in situ and cancer in diploid cases (p=0.0002), and further at the change from diploid to aneuploid (p=0.004). At a median cancer specific survival time of four months patients with diploid tumours had a better survival than those with aneuploid tumours (p=0.02). In multivariate analysis of the tumour characteristic, only T-category and tumour grade were independent prognostic factors. The shift from diploid to aneuploid and the further shift of ploidy within aneuploid tumours are in agreement with the concept of a clonal development of gallbladder cancer. These changes are combined with a stepwise increase in the fraction of S-phase cells. Low frequency of symptoms in single stone patients may be the reason for detection of malignancy at a late stage of tumour development. PMID- 12082296 TI - Automated detection of connective tissue by tissue counter analysis and classification and regression trees. AB - OBJECTIVE: To evaluate the feasibility of the CART (Classification and Regression Tree) procedure for the recognition of microscopic structures in tissue counter analysis. METHODS: Digital microscopic images of H & E; stained slides of normal human skin and of primary malignant melanoma were overlayed with regularly distributed square measuring masks (elements) and grey value, texture and colour features within each mask were recorded. In the learning set, elements were interactively labeled as representing either connective tissue of the reticular dermis, other tissue components or background. Subsequently, CART models were based on these data sets. RESULTS: Implementation of the CART classification rules into the image analysis program showed that in an independent test set 94.1% of elements classified as connective tissue of the reticular dermis were correctly labeled. Automated measurements of the total amount of tissue and of the amount of connective tissue within a slide showed high reproducibility (r=0.97 and r=0.94, respectively; p<0.001). CONCLUSIONS: CART procedure in tissue counter analysis yields simple and reproducible classification rules for tissue elements. PMID- 12082297 TI - Automatic registration and error detection of multiple slices using landmarks. AB - OBJECTIVES: When analysing the 3D structure of tissue, serial sectioning and staining of the resulting slices is sometimes the preferred option. This leads to severe registration problems. In this paper, a method for automatic registration and error detection of slices using landmark needles has been developed. A cost function takes some parameters from the current state of the problem to be solved as input and gives a quality of the current solution as output. The cost function used in this paper, is based on a model of the slices and the landmark needles. The method has been used to register slices of prostates in order to create 3D computer models. Manual registration of the same prostates has been undertaken and compared with the results from the algorithm. METHODS: Prostates from sixteen men who underwent radical prostatectomy were formalin fixed with landmark needles, sliced and the slices were computer reconstructed. The cost function takes rotation and translation for each prostate slice, as well as slope and offset for each landmark needle as input. The current quality of fit of the model, using the input parameters given, is returned. The function takes the built-in instability of the model into account. The method uses a standard algorithm to optimize the prostate slice positions. To verify the result, s standard method in statistics was used. RESULTS: The methods were evaluated for 16 prostates. When testing blindly, a physician could not determine whether the registration shown to him were created by the automated method described in this paper, or manually by an expert, except in one out of 16 cases. Visual inspection and analysis of the outlier confirmed that the input data had been deformed. The automatic detection of erroneous slices marked a few slices, including the outlier, as suspicious. CONCLUSIONS: The model based registration performs better than traditional simple slice-wise registration. In the case of prostate slice registration, other aspects, such as the physical slicing method used, may be more important to the final result than the selection of registration method to use. PMID- 12082298 TI - A field observation on color selection by New World sympatric primates, Pithecia pithecia and Alouatta seniculus. AB - This work characterizes differences in selection of Talisia retusa fruits by two sympatric Neotropical primates, Pithecia pithecia (white-faced sakis) and Alouatta seniculus (red howlers). Color appears to be the criterion by which fruits were selected. Greenish fruits were mainly eaten by Pithecia, while yellowish fruits by Alouatta. The characteristics of these primates in relation to seed predation and seed dispersal are discussed in the context of the Talisia retusa fruit color spectrum. Furthermore, a possible differential acquisition of chemical components, like tannins, is hypothetically treated considering the variation in fruit color. PMID- 12082299 TI - Behavioral seasonality in Mahale chimpanzees. AB - To analyze how the chimpanzees in the Mahale Mountains National Park, Tanzania, change their grouping pattern, activity budget, travel speed, and travel distance within an annual cycle, I divided 1-year data into four periods. The Mahale chimpanzees have the behavioral flexibility to adapt to various climates and exhibited at least three behavioral seasons. In the early wet season, chimpanzees formed a few, large parties, and spent much time feeding on insects and animal meat. In the early and late dry seasons, chimpanzees maintained party sizes as large as in the early wet season, and traveled distances similar to the early wet season, but spent the most time feeding and traveling within the year. By contrast, in the late wet season chimpanzee parties broke up into more numerous, small groups, and traveled slowly over shorter distances. Although time spent feeding and traveling were the same as that in the early wet season, time spent feeding on terrestrial herbaceous vegetation (THV) was the highest in the year. The results suggest that chimpanzees travel longer, faster, and farther in seasons when they form large parties. PMID- 12082300 TI - Intergroup encounters in wild moor macaques (Macaca maurus). AB - We studied intergroup encounters among moor macaques at the Karaenta Nature Reserve, South Sulawesi, Indonesia. Group B has been observed on the basis of individual identification since 1988. We analyzed 85 encounters between members of Group B and members of neighboring groups from September 1990 to November 1998. The average frequency of intergroup encounters was 0.035/hour. Neither the presence of females in estrus nor rainfall had an effect on encounter frequency. Behaviors of moor macaques during intergroup encounters differed from those of Japanese macaques. In moor macaques, no intergroup interactions with body contact were observed during encounters, and females never directed aggression toward members of different groups. The present study did not confirm the prediction of the model of van Schaik (1989). Extension of the existing models is required to explain the difference in female dominance styles among macaques by socioecological factors. PMID- 12082301 TI - The finding of an inverse relationship between social dominance and feeding priority among pairs of unfamiliar adult male vervet monkeys (Cercopithecus aethiops sabaeus). AB - Dominance is often presumed to confer priority of access to resources. This study evaluated the relationship between two assessments of dominance: (1) social dominance, based on agonistic interactions and (2) feeding priority among pairs of unfamiliar adult vervet monkeys (Cercopithecus aethiops sabaeus) differing in scrotal colour, but matched for height, weight and testicular volume, during paired introduction experiments. Results of this investigation showed that neither size differences nor scrotal colour were predictive of feeding priority, and social dominance was inversely related to feeding priority. This finding demonstrates that different assessments of dominance can yield different outcomes even within the same primate taxon. I propose that male dominance rank may best predict access to resources when there is direct contest competition over a resource, which is not immediately exhaustible, whereas highly impulsive low ranking males may gain a competitive edge in scramble competitions for ephemeral and small resources. PMID- 12082302 TI - Changes of dominance rank, age, and tenure of wild Japanese macaque males in the Kinkazan a troop during seven years. AB - Male age-rank and tenure-rank relationships were studied for seven years in unprovisioned Japanese macaque (Macaca fuscata fuscata) troop on Kinkazan Island, Japan. Males whose estimated ages were between 15 and 19 yr old monopolized the highest ranks, while older males whose estimated ages were > or = 20 yr old tended to decline in rank, resulting in a humped age-rank curve. The ranks of males tended to rise as their tenure in the troop increased. The departure of higher-ranking males was the social mechanism for changes in rank, suggesting that the disappearance of higher-ranking males plays an important role in determining rank dominance. PMID- 12082303 TI - A preliminary study on the social relationships in a semi-free ranging colony of sun-tailed monkeys (Cercopithecus solatus), a species recently discovered in Gabon. AB - In 1984, a species of guenon endemic to Gabon was discovered: the sun-tailed monkey (Cercopithecus solatus). This species is difficult to locate and observe in the wild, and hence to date has been little studied. The Centre International de Recherches Medicales de Franceville (CIRMF), Gabon, houses the world's only breeding colony of C. solatus, on which eco-ethological investigations can be carried out in a semi-free ranging environment. The data reported here present the first results of observations on the social relationships of this colony and support the scant field observations available on this species, showing a basic social unit of one adult male and several females with their offspring. The resident male systematically repulses any second adult male in proximity to the group. A clear hierarchy exists among the females, with mature female offspring eventually acquiring a rank just below that of their mother. PMID- 12082304 TI - The relationship between tail use and positional behavior in Alouatta palliata. AB - The relationship between tail use and positional behavior is explored in Alouatta palliata. During bridging, climbing, suspension, standing, and sprawling, the tail is attached to a substrate for the majority of sample points. Tail attachment was more likely to occur when the animal is traveling on vertical or terminal substrates. Quadrupedalism showed few instances of attachment and sitting reflected nearly equal amounts of prehension and non-prehension. Tail prehension is used in all behavioral contexts but shows higher frequencies of attachment during feeding than during resting, or foraging. Tail prehension appears to aid in the stability, support, and balance of the animal across numerous positional behaviors. PMID- 12082305 TI - Discovery of immunostimulatory CpG-DNA and its application to tuberculosis vaccine development. AB - DNA containing an unmethylated CpG motif has a potent immunostimulatory effect on the vertebrate immune system. Because such CpG motifs are relatively common in bacterial DNA, but rare in mammalian animal and plant DNA, they may be an evolutionary adaptation augmenting innate immunity, most likely in response to pathogens that replicate within the host cells, such as viruses and intracellular bacteria. Microbial infection induces innate immunity by triggering pattern recognition systems. The infected cells produce proinflammatory cytokines that directly combat microbial invaders and express costimulating surface molecules, which develop adaptive immunity by inducing distinct T cell differentiation. Bacterial DNA with unmethylated CpG-DNA stimulates vertebrate immature immune cells to induce maturation and to produce TNF-alpha as well as Th1-type cytokines, IL-12 and IFN-gamma. Therefore, CpG-DNA functions as an adjuvant for regulating the initiation of Th1 differentiation. The roles of immunostimulatory CpG motifs in DNA vaccine developments and in therapeutic applications have been discussed. PMID- 12082306 TI - Antibiotic susceptibility pattern of Salmonella worthington isolated from neonates--a retrospective study. AB - Salmonella worthington is an emerging pathogen and has been implicated in a number of outbreaks of neonatal meningitis and septicemia. Over a period of 5 years, a total of 30 strains of this pathogen were isolated from blood and cerebrospinal fluid of neonates suffering from septicemia with or without meningitis. Most of these strains were resistant to the penicillin group of antibiotics, and many were resistant to cefotaxime. Sixty percent of the isolates were resistant to amikacin; 86% were resistant to chloramphenicol, and none were resistant to ciprofloxacin or norfloxacin. Parenteral fluoroquinolone should be included as part of antibiotic therapy in suspected cases of neonatal meningitis due to S. worthington. PMID- 12082307 TI - Subconjunctival infection with Dirofilaria repens. AB - Cases of zoonotic dirofilariasis infection, caused by Dirofilaria repens, occur widely throughout European, African, Middle Eastern, and Asian countries. The reports of this infection in humans in India are limited, and we herein report the case of a 23-year-old man who presented with subconjunctival swelling in the right eye with no local symptoms. A large nematode was visualized on examination and the intact worm was surgically removed. The parasite was identified as a male D. repens. PMID- 12082308 TI - Chlamydophila pneumoniae specific antibodies in Thai patients with myocardial infarction. AB - To investigate the correlation between Chlamydia pneumoniae infection and acute myocardial infarction (AMI), a total of 101 serum specimens collected from patients with AMI admitted to the coronary care unit, Bhumibol Adulyadej Hospital, and serum specimens collected from healthy blood donors (control group) were examined by using the micro-immunofluorescence test. C. pneumoniae antibody positive cases were found in 52 (52%) patients, consisting of 30 males and 22 females, though no significant difference of prevalence rate was observed when compared with the rate in the control group. However, the level of IgG titers in patients was higher than that in the controls, and this finding may support an association between C. pneumoniae infection and AMI. Among patients with AMI, several cases were suspected to have current infections because of a fourfold or higher titer increase in IgG or titers in IgM antibody of 1:32 or 1:64. There is no significant correlation between serologic test results and diabetes mellitus, hypertension, hyper cholesterol, a smoking habit, or the location of myocardial infarction among patients with AMI. PMID- 12082309 TI - Diphtheria immunity of different age groups in Turkey. AB - In Turkey, the immunization policy against diphtheria involves childhood vaccination, and no routine booster dose is administered after 12 years of age. Information about the immune status of the population is important to predict the potential for epidemics, several of which have occurred recently in neighboring countries. The aim of this study was to assess the immune status of different age groups in Turkey against diphtheria. A total of 497 blood samples were collected, and diphtheria antitoxin levels were measured by the Enzyme Linked Immunosorbent Assay method. Titers below 0.1 IU/ml were considered to show insufficient immunity. Overall, 35.8% of the population studied showed insufficient immunity against diphtheria. Immunity rates varied significantly among different age groups (P < 0.01). Protective immunity rates were highest in the 3- to 6-year age group (81.3%) and the over 60 age group (81.2%). In subjects between the ages of 10 and 59 the percentage of people with insufficient immunity was above 30%. The lowest immunity rate was observed in 20- to 29-year age group. No significant difference was seen between males and females in terms of protective effect of vaccination. This study indicates that people between the ages of 10 and 59 are susceptible to diphtheria in our country. Therefore, revaccination of adults with tetanus-diphtheria toxoids at every opportunity (military service, pregnancy, admittance to emergency room) should be considered as an immunization policy. PMID- 12082310 TI - Tick-borne encephalitis virus within and outside Japan: a cause for concern? PMID- 12082311 TI - Surveillance of poliovirus-isolates in Japan, 2001. PMID- 12082312 TI - Antimicrobial resistance pattern of Salmonella typhi isolates in Kolkata, India during 1991-2001: a retrospective study. PMID- 12082313 TI - Induction of HIV-1-specific neutralizing antibodies in mice vaccinated with a recombinant Sendai virus vector. PMID- 12082314 TI - Reproducibility of oral bacterial isolation in the elderly. PMID- 12082315 TI - Discordant movement of CD4-positive T-cell count in HIV-1 infected patients with HAART failure. PMID- 12082316 TI - Is there a relation between the antibiotic sensitivity of methicillin- resistant Staphylococcus aureus and the consumption of antibiotics in a hospital?: a study from April 1993-March 1999. PMID- 12082317 TI - Antiphospholipid syndrome and recurrent miscarriage. PMID- 12082318 TI - Beta2-glycoprotein I dependent anticardiolipin antibodies and lupus anticoagulant in patients with recurrent pregnancy loss. AB - AIM: The present study was aimed to define the incidence of antiphospholipid antibodies of different types lupus anticoagulant (LAC), venereal disease research laboratory test (VDRL) and Beta2-glycoprotein I dependent anticardiolipin antibodies Beta2 I aCL) in our cohort of population experiencing recurrent pregnancy loss (RPL) from Andhra Pradesh, South India. SETTING AND DESIGN: A referral case-control study at a tertiary centre over a period of 5 years. PARTICIPANTS: 150 couples experiencing 3 or more recurrent pregnancy losses with similar number of matched controls. MATERIAL AND METHODS: LAC activity was measured by the activated partial thromboplastin time (aPTT) according to the method of Proctor and Rapaport with relevant modifications. VDRL analysis was performed by the kit method supplied by Ranbaxy Diagnostics Limited and Beta2 Glycoprotein I dependent anticardiolipin antibodies were estimated by ELISA kit (ORGen Tech, GmbH, Germany) with human Beta2 Glycoprotein I as co factor. STATISTICAL ANALYSIS: Statistical analysis was performed using Student's t test. RESULTS: LAC activity was found positive in 11 women (10.28%). The mean +/- SE Beta2 I aCL concentration in the study group was 14.53 (micro/ml) +/- 1.79 (range 0 to 90.4 micro/ml) which was higher than the control group with a mean +/ SE of 7.26 (micro/ml) +/- 0.40 (range 0 to 18 u/ml). The binding of the antibodies to the antigen was observed in 40.24% (n=33) of the cases compared to 6.09% (n=5) in controls. VDRL test was positive in 7(2.34%) individuals (3 couples and 1 male partner) and none among controls. CONCLUSIONS: The present study indicates the importance of antiphospholipid antibodies in women experiencing RPL and suggests the usefulness of screening for these antibodies as a mandatory routine for instituting efficient therapeutic regimens for a successful outcome of pregnancy. PMID- 12082319 TI - Laryngeal mask airway vs endotracheal tube to facilitate bedside percutaneous tracheostomy in critically ill patients: a prospective comparative study. AB - BACKGROUND: Inadvertent puncture of endotracheal tube (ETT) cuff and accidental tracheal extubation are the potential life threatening complications during percutaneous dilatational tracheostomy (PDT). As an alternative to ETT, the laryngeal mask airway (LMA) has been used sporadically but no large study is available on its use. AIMS: To study the use of LMA during PDT on controlled ventilation in critically sick patients and compare its advantages and disadvantages in relation to ETT. SETTINGS AND DESIGN: This prospective, randomised, comparative study was conducted in intensive care unit of a super speciality, tertiary care hospital. METHODS AND MATERIAL: The bedside PDT was performed in 60 critically ill patients using Ciaglia's kit. The patients received controlled ventilation of lungs either through ETT or LMA. The feasibility of the procedure and complications were compared in two groups. RESULTS: In LMA group, 33% of patients suffered with potentially catastrophic complications, e.g., loss of airway, inadequate ventilation of lungs leading to significant hypoxia, gastric distension and regurgitation. In ETT group there were 6.6% incidence of ETT impalement, 6.6% cuff puncture and 3.3% accidental tracheal extubation. Despite the technical difficulties (cuff puncture, etc.) the ETT technique was basically safe, whereas despite of the theoretical advantages the LMA technique rendered some patients at considerable risks of hypoxia and gastric regurgitation-aspiration. CONCLUSION: The LMA does not provide safe patent airway to facilitate bedside PDT in critically sick population on controlled ventilation. The ETT is safer for controlled ventilation and should be continued to secure the airway for this purpose until a better alternative is available. PMID- 12082320 TI - Development of a distress inventory for cancer: preliminary results. AB - CONTEXT: Advances in cancer treatment have led to cure and prolongation of patients' lives; however associated psychosocial problems, including distress, can detrimentally affect patients' compliance with treatment and ultimately, their outcome. Symptom distress has been well addressed in many studies; however, psychological distress has only been quantified by using depression or anxiety scales/checklists or quality of life scales containing a distress sub scale/component or by the use of scales that are not psychological distress specific. AIMS: The present study is an attempt to construct a psychological distress inventory for specific use with cancer patients. SETTINGS AND DESIGN: The standardisation sample consisted of 63 randomly selected patients with head and neck cancer who had undergone/ were undergoing curative treatment at the Regional Cancer Centre, Trivandrum. PATIENTS AND METHODS: The Distress Inventory for Cancer contained 57 positively and negatively toned items. An item analysis was conducted, followed by a factor analysis, thereby identifying the domains influencing distress. RESULTS: The final questionnaire contained 26 items subdivided into four domains viz. the personal, spiritual, physical, and the family domains, with each domain providing a sub score. The reliability coefficient (Cronbach's alpha) of the scale was found to be 0.85. CONCLUSIONS: These are the preliminary results of an ongoing study on global distress and tool development process. Reported here is the first step towards development of such tool. PMID- 12082321 TI - An analysis of long-term venous access catheters in cancer patients:experience from a tertiary care centre in India. AB - BACKGROUND: Venous access is crucial for cancer management for administration of drugs blood products, antibiotics and periodic sampling. AIMS: To review our experience of long-term venous access devices used over a ten-year period and to analyse the outcome in cancer patients in Indian setting. SETTING AND DESIGN: A retrospective analysis of data in a tertiary care Regional cancer centre. PATIENTS AND METHODS: A total of 110 patients with various malignancies requiring long-term venous access were included in the study. A uniform open cut down procedure under local anaesthesia was used and silastic Hickman catheters were inserted in the cephalic or external jugular or internal jugular veins. A record of all complications and catheter loss and final out come were analysed. RESULTS: A total of 111 catheters were used in 110 patients. Sixty-nine catheters were placed in cephalic, 40 in external jugular, and 2 in internal jugular vein. Duration of catheter indwelling period ranged from 7 to 365 days with a median of 120 days. In 90% of the cases the catheter tip was located either in superior vena cava or in right atrium. Total catheter related complications were observed in 37 (34.54%) patients and catheter loss rate due to complications was 15.4% (17/111). CONCLUSIONS: Long-term venous access using Hickman catheter insertion by open cut down method is a simple, safe and reliable method for administration of chemotherapeutic agents, antibiotics and blood products. The incidence of various complications and catheter loss was acceptable and overall patient satisfaction was good. PMID- 12082322 TI - Laparoscopic orchidectomy for undescended testis in adults. AB - BACKGROUND: Impalpable testis is a significant diagnostic and therapeutic challenge in adults, for both radiologist and surgeons, with few reports in literature addressing this problem in adults. Laparoscopy is a reliable and definitive procedure obviating the necessity of advance investigation and subsequent inguinal exploration in adults. AIMS: To study the utility of laparoscopy as combined diagnostic and therapeutic modality for undescended testis in adults. SETTINGS AND DESIGN: Prospective study from a single surgical unit of a large tertiary referral centre during August 2000 to January 2002. METHODS AND MATERIAL: Nine patients of unilateral undescended testis with average age 22.7 years (range 13-31 years) underwent diagnostic laparoscopy and orchidectomy subsequent to detailed clinical, ultrasound and examination under anaesthesia (EUA) procedure. All patients were operated with one 10 mm umbilical camera port, one suprapubic port and 1 lateral port. RESULTS: None of the patients had palpable testis or an inguinal cough impulse on clinical examination and during EUA. In only 3(33.3%) patients, the ultrasound could locate the testis situated at the deep ring. On laparoscopy all testes were identified, 4 were present at the deep ring, 3 were intra-abdominal and 2 had blind ending vas entering the deep ring. Mesh plug was inserted in the internal ring in these 2 patients, after dissecting the peritoneum. None of the patients had intra or post operative complications and all were discharged on the next day. CONCLUSION: Laparoscopy is one of the most satisfactory methods for the diagnosis and management of non-palpable testis in adult cryptorchid patients. PMID- 12082323 TI - Correlation of Helicobacter pylori and gastric carcinoma. AB - BACKGROUND: Difference of opinion about the prevalence of H. pylori association with gastric cancer exists in the literature. AIMS: To study the correlation of Helicobacter pylori (H. pylori) to gastric carcinoma. METHODS: 50 proved cases of gastric cancer were studied by rapid urease test, culture, histopathology and ELISA test for H. pylori IgG. RESULTS: 68% of cases of gastric cancer were found to be positive for H. pylori infection as compared to 74% of healthy controls. CONCLUSIONS: The prevalence rate of H. pylori infection in our patients of gastric cancer was lower than in the control population though statistically not significant, suggesting that H. pylori may not be responsible for gastric carcinogenesis in this population. PMID- 12082324 TI - Primary pleuro-pulmonary malignant germ cell tumours. AB - Lungs and pleura are rare sites for malignant germ-cell tumours. Two cases, pure yolk-sac tumour and yolk sac-sac tumour/embryonal carcinoma are described in young males who presented with rapid progression of respiratory symptoms. The malignant mixed germ cell tumour occurred in the right lung, while the yolk-sac tumour had a pseudomesotheliomatous growth pattern suggesting a pleural origin. Alpha-foetoprotein was immunohistochemically demonstrated in both. PMID- 12082325 TI - Cutaneous metastasis from carcinoma of tonsil. AB - Hematogenous spread from carcinoma of tonsil is an uncommon event and skin is an extremely rare site of metastasis. We encountered a 40-year-old male patient who initially presented with carcinoma of the tonsil with T3N2cMO disease and treated by curative radiotherapy. After about 2 years, he developed a skin lesion in the periorbital region which on cytological examination turned out to be metastasis from tonsillar carcinoma. The present paper describes this rare case report along with a brief review of the literature. PMID- 12082326 TI - Pressure controlled inverse ratio ventilation in acute respiratory distress syndrome patients. AB - Appropriate ventilatory intervention is life saving in acute respiratory distress syndrome (ARDS). Pressure controlled inverse ratio ventilation (PC-IRV) is the likely mode of ventilation benefiting in extreme conditions of ARDS. However, guidelines when to start PC-IRV is not yet well defined. The ventilation-related dilemma, which we faced in two illustrative cases of ARDS are presented. The first patient presenting clinically with ARDS but with high peak airway pressure (PIP) and low dynamic lung compliance, PC-IRV helped in reducing PIP, improved haemodynamics and the oxygenation of blood. In second patient with similar clinical presentation of ARDS, where although PIP was high but the dynamic compliance was better, the PC-IRV caused deterioration in PaO2. Here, patient rather did better with high PEEP (15 cm H2O) and usual I: E ratio (1:2). It is probable that the dynamic lung compliance (< 20ml/cmH2O), PIP (> 50 cm H2O) at conventional I: E ratio (1:2) ventilation (10 ml/kg) with hypotension might form the basis to develop a scoring system for guidance to switch over to PC-IRV ventilation. Further randomised prospective controlled clinical trials will then be required to establish indication to start PC-IRV in ARDS. PMID- 12082327 TI - Acquired pure red cell aplasia in a child. AB - Primary acquired pure red cell aplasia is a rare occurrence in childhood. An eleven-year old boy presented to us with pallor, which required multiple packed red cell transfusions. He did not have hepatosplenomegaly, jaundice or lymphadenopathy. Bone marrow examination revealed the diagnosis of pure red cell aplasia. All possible investigations were done to exclude secondary causes of pure red cell aplasia. No secondary cause was found on investigations. Rheumatoid factor and anti-nuclear antibodies were positive. He was started on oral steroids, to which he did not respond. He was then given cyclosporine A. Response to cyclosporine was dramatic and the child now does not require any transfusions. PMID- 12082328 TI - Laparoscopy, robot, telesurgery and urology: future perspective. PMID- 12082330 TI - Clinical signs in medicine: pulsus paradoxus. PMID- 12082331 TI - Robinow syndrome. PMID- 12082332 TI - Necrotising histiocytic lymphadenitis. PMID- 12082333 TI - Intramedullary tuberculoma of the spinal cord. PMID- 12082334 TI - Allograft biopsies in management of pancreas transplant recipients. AB - Pancreatic transplantation is becoming increasingly accepted as a treatment modality for Type 1 diabetes mellitus. When allograft dysfunction is noted during follow up of patients, a biopsy is an extremely useful tool to diagnose various forms of rejection, and to rule out non-immunologic causes of graft malfunction, such as donor disease, ischaemic/preservation injury, vascular thrombosis, pancreatitis, post-transplant lymphoproliferative disease, technical complications and recurrence of diabetes mellitus. In addition to its role in establishing the primary diagnosis, a biopsy can grade the severity of pathology present, help determine the most appropriate therapy, and provide information relevant to graft prognosis. PMID- 12082336 TI - Chromosome 19; 20 reciprocal translocation. PMID- 12082335 TI - Prevention of postoperative acute renal failure. AB - Postoperative acute renal failure (PO-ARF) is a serious complication resulting in a prolonged stay and high mortality. Patients may be at risk for this problem because of an underlying medical illness, nature of surgery, nephrotoxin exposure, or combinations of these factors. An increase in the intra abdominal pressure above 20-mm Hg is associated with an increase in the incidence of PO ARF. Based on many clinical studies in high-risk surgical patients and patients undergoing renal transplantation, the only proven management strategies for prevention of PO-ARF are adequate volume expansion and avoidance of hypovolaemia. Drugs known to be nephrotoxic should be avoided or used with caution. Three main pharmacological agents namely mannitol, frusemide and dopamine have been extensively tried in the prevention of PO-ARF. Mannitol has proven of value only in the presence of adequate volume expansion in attenuating renal dysfunction in transplant patients. Frusemide converts oliguric renal failure to non-oliguric renal failure. The bulk of the data, including that from prospective studies indicate dopamine is only a diuretic. Fenoldopam, a dopamine analogue, has shown early promise in reports. Calcium channel blockers have not been shown to improve the outcome in renal transplantation or help in the prevention of contrast induced nephropathy. Atrial natriuretic peptide has not been proven to be of benefit in established renal failure and its role in prevention has not been assessed. PMID- 12082337 TI - Anterior transverse cervical rupture following intra-amniotic prostaglandin induced mid-trimester abortion. PMID- 12082338 TI - Isolated gastric tuberculosis presenting as haematemesis. PMID- 12082339 TI - History of plastic surgery in India. PMID- 12082340 TI - The man who saved us all. PMID- 12082341 TI - Rabeprazole. PMID- 12082342 TI - Structure determination of proteins in solution by nuclear magnetic resonance spectroscopy. PMID- 12082343 TI - [Outpatient parenteral antibiotic treatment: the Italian model]. AB - Though constrained by the lack of standard criteria for patient selection and well defined therapeutic protocols, the Italian model of OPAT is characterized by the home treatment of infections also of moderate severity, chiefly of the lower respiratory tract by means of broad spectrum antibiotics (chiefly beta-lactams) administered intramuscularly. The selection of patient for OPAT remains an individual decision of GPs based on the severity of infection, concomitant diseases, age of the patient, reliability of patients and family, environ- mental and social factors. Such a model has gained broad consensus over the years from patients for different cultural reasons: first, the willingness of patients to participate in the OPAT programmes, second, the concept that home is better than hospital even when you are ill and, third, the fact that parenteral drugs are perceived as more effective than oral ones, probably in contrast with the culture and traditions of other countries (especially northern Europe) where hospitals are still considered the only proper site of care and intramuscular administration is regarded as almost unnatural. No private or insurance reimbursement based OPAT structure exists up to now in Italy or is go. ing to be developed. The main problems in Italy now seems to be: -to define clear protocols and easy criteria for selection of patients enable to OPAT, -to improve what existing by creating and/or qualifying new professionals involved with in- and outpatients (OPAT, early discharge,medical care for special populations.). PMID- 12082344 TI - [Vascular graft infection by Staphylococcus epidermidis: efficacy of various perioperative prophylaxis protocols in an animal model]. AB - A rat model was used to investigate the efficacy of levofloxacin, cefazolin and teicoplanin in the prevention of vascular prosthetic graft infection. Graft infections were established in the subcutaneous tissue of 300 male Wistar rats by implantation of Dacron prostheses followed by topical inoculation with methicillin-susceptible and methicillin-resistant S. epidermidis. The study included a group without contamination, two contaminated groups without prophylaxis, two contaminated groups with intraperitoneal levofloxacin prophylaxis (10 mg/kg), two contaminated groups with intraperitoneal cefazolin prophylaxis (30 mg/kg), two contaminated groups with intraperitoneal teicoplanin prophylaxis (10 mg/kg) and six contaminated groups with rifampin-soaked graft and intraperitoneal levofloxacin, cefazolin or te- icoplanin prophylaxis. The grafts were removed after 7 days and evaluated by quantitative culture. The efficacy of levofloxacin against the methicillin- susceptible strain did not differ from that of cefazolin or teicoplanin. Levofloxacin showed slight less efficacy than teicoplanin against the methicillin-resistant strain. The levofloxacin-rifampin combination proved to be similarly effective to the rifampin-teicoplanin combination and more effective than the rifampin-cefazolin combination against both strains. The rifampin-levofloxacin combination may be useful for the prevention of late-appearing vascular graft infections caused by S. epidermidis because it takes advantage of the good anti-staphylococcal activity of both drugs. PMID- 12082345 TI - [Microbial environmental surveillance in a bone marrow transplant unit]. AB - The aim of this study was to evaluate, after several years of application, in a bone marrow transplant (BMT) center of a Cagliari Hospital, the effectiveness of the disinfection protocol in minimizing the risk of environmentally transmitted infections. Microbial contamination of the air was evaluated every two months during normal activity using an SAS sampler. The contamination of surfaces was determined weekly, 2hrs after sanitation, using disposable surface contact plates. The results of environmental monitoring generally showed low values of microbial contamination of air and surfaces. Only two service rooms and two patient's rooms without own bathroom showed levels of microbial contamination slightly exceeding, in a few samplings, the values considered acceptable for environments at high risk of infections. From a qualitative point of view, the microrganisms isolated generally belonged to environmental species. In conclusion our study confirms the importance of microbial monitoring in the control and prevention of outbreaks of infections in BMT Units. This approach allows significant reduction in the level of contamination not only by improving cleaning procedures, but also by motivating the cleaning staff trough making them aware of their responsibilities. PMID- 12082346 TI - [Survey of disinfectant usage in a polyclinic in Rome: preliminary results]. AB - The authors describe the survey that was conducted in the largest hospital in Rome. The aim was to estimate the disinfectants and chemical sterilizers employed and asses possible hazards for patients and workers arising from the use of disinfectants, together with the problem of waste disposal. The study was conducted by means of a survey form that was distributed to 39 departments/institute in the whole hospital of which 24 (61%) responded. Overall 635 charts were collected listing 886 usage procedures and 110 formulations with 40 different active ingredients. The disinfection fields of use were: skin(52.4%) environment (28.8%) and hospital equipment (20.8%) This study shows that a modern hospital should select very few disinfectants and prepare a handbook that can explain disinfectant usage procedures and concentrations. In the guidelines the problem of waste disposal and employee safety should also be considered. PMID- 12082347 TI - [Clonal diffusion and evolution of mecA and Tn554 polymorphisms in methicillin resistant Staphylococcus aureus in Italy]. AB - The purposes of the present study were to track the geographic spread of 69 MRSA strains in Italy recovered from 7 hospitals in four towns; to detect the clonal identities among the isolates by a combination of multiple genomic typing methods and to measure temporal trends in clonal types between 1984 and 1998. Our results showed the spread of three major clones among the MRSA isolates of 1984-1995 period: the most represented MRSA clone carried the PFGE pattern A, the mecA polymorph II and had no homology with Tn554 (II::NH::A); most of these isolates were susceptible to the macrolides,being similar to the historically " archaic" MRSA strains; the clone typed I::E::A, carried the PFGE pattern A, the mecA polymorph I and Tn554E commonly defined as "Iberian clone"; unique clone, showing an uncommon PFGE pattern E. the mecA polymorph II and the Tn554 E (II::E::E) and were characterized by a uniform susceptibility to tetracycline and rifampin. During 1997-98 the representation of this clone increased instead of the classical "Iberian clone". A new multi-resistant MRSA strain, carrying the PFGE pattern B (or B'), the mecA polymorph XI and Tn554 polymorph B (XI::B::B), called "Brazilian clone", increased from being absent (1984-95) to 48%. Our molecular data show an Italian MRSA "scenario" far from the common European trends and clearly documented the spread of an archaic clonal type (II::NH::A) in 1984-95, the arrival and rapid increase of Brazilian done in 1997-98 and the chronological and geographical spread of a unique (H::E::E) called "Italian clone", instead of the widely spread Iberian MRSA clone. PMID- 12082348 TI - Transverse myelitis associated with Mycoplasma pneumoniae pneumonitis: a report of two cases. AB - The authors report two cases of transverse myelitis due to Mycoplasma pneumoniae occurring during a recent acute infection due to Epstein-Barr virus (EBV). The clinical picture included weakness of the legs, bladder dysfunction, fever, headache and in one case, a mild confusional state. Magnetic Resonance Imaging (MRI) revealed a segmental edema of the lumbar tract and cerebro-spinal fluid analysis showed a mild pleocytosis and an increased level of proteins with a blood-brain barrier damage. The diagnosis of acute infection due to Mycoplasma pneumoniae was made on serological criteria. The patients recovered completely after a 14-day course of an associated regimen with Ciprofloxacin (400 mg/day i.v.) plus prednisone (60 mg/day i.v.). The authors emphasise the immunological disorders in the combined infection with Mycoplasma pneumoniae and EBV, supposing that an infective mononucleosis may predispose to mycoplasmic transverse myelitis. PMID- 12082349 TI - [Pathogenic role of Acinetobacter spp during HIV infection]. AB - In order to assess the clinical role of bacterial complications due to Acinetobacter spp. during HIV disease, a retrospective survey of clinical and microbiological data of 2221 HIV-infected patients hospitalised during the past 10 years was carried out, evaluating all episodes of Acinetobacter spp. infection according to several epidemiological, clinical and therapeutic variables. Eleven patients of 2221 (0.5%) suffered from Acinetobacter spp. disease: sepsis in 5 cases, and urinary, respiratory tract disease and bacteremic pneumonia in three, two, and one patient respectively. A. calcoaceticus was responsible in 4 cases, A. lwoffii in three, and Acinetobacter spp. in the 4 remaining cases; 4 patients experienced a polymicrobial infection, and 7 had a prior diagnosis of full-blown AIDS. All patients had a severe HIV-related immunodeficiency (mean CD4+ lymphocyte count 118.2 +/- 45.3 cells/microl). Compared with other localizations, sepsis was related to a lower mean CD4+ cell count (p<.001), and a more frequent occurrence of leucopenia-neutropenia (p<.005). Disease episodes diagnosed after the first 72 hours of hospitalisation (deemed no- socomial in origin), proved more frequent than community-acquired ones (9 cases versus 2), affected predominantly patients with AIDS and neutropenia, and were frequent1y associated with bacteremia (p<.04) The use of broad spectrum antibiotics, corticosteroids and cotrimoxazole, was recognized during the month preceding the diagnosis of Acinetobacter spp. disease, in 6, 4, and 8 cases, respectively. One patient only had an indwelling intravascular catheter, while no recent history of surgery, intensive care, or other invasive procedures was found. At in vitro susceptibility studies, bacterial isolates showed complete resistance to ampicillin and cephalothin, and low sensitivity to second-generation cephalosporins, while a higher susceptibility rate was revealed towards ceftazidime, netilmicin, amikacin, and quinolones, followed by cotrimoxazole and piperacillin. A prompt and appropriate antimicrobial therapy (mostly carried out with cephalosporins and aminoglycosides), led all patients to a clinical and microbiological cure within 6-1.3 days, in the absence of mortality or relapses. As opportunist pathogens with a predominant nosocomial origin, Acinetobacter spp. organisms may be responsible for an appreciable morbidity in patients with HIV disease, especially when additional risk factors (immunodeficiency, underlying diseases, and hospitalisation) are present. Notwithstanding the high drug resistance profile of the majority of isolated organisms, a timely diagnosis and a treatment based on in vitro assays, contribute to avoid recurrences and potentially life-threatening complications. PMID- 12082350 TI - [Innovation in nineteenth century healthcare: disinfection stations]. AB - The possibility of limiting and preventing epidemics by cleansing everything that could be touched by patients, was an intuitive thought well before microbial discoveries. In accordance with the "miasmas theory", morbid substances were emanated from the bodies of the patients. A step forward took place in the 19th century with Pasteur and Koch's discoveries and progress in chemical disinfectants. The law was slow to adapt: in Italy only in 1888 was a Code drawn up to establish some "Disinfection Stations", places for the sterilisation of infectious material from patients' homes. Later, a similar home service was started for everything that was not transportable. Thus, in the case of cholera, smallpox, scarlet fever, diphtheria, typhus, tuberculosis, etc. it became possible to sterilise everything with which patients came into contact. PMID- 12082351 TI - Welcome to Travel Nursing 2002. PMID- 12082352 TI - Home sweet home. PMID- 12082353 TI - What to see and do in San Francisco. PMID- 12082354 TI - What to see and do in Orlando/Lake Buena Vista. PMID- 12082355 TI - What to see and do in Dallas. PMID- 12082356 TI - HRT and cognitive function: what are we to believe? PMID- 12082357 TI - Add-backs to prevent skeletal fragility: foresight or folly? PMID- 12082358 TI - Cognitive function effects of suppressing ovarian hormones in young women. AB - OBJECTIVE: To evaluate the effect of a pharmacologically induced, temporary suppression of ovarian hormones on healthy young women's cognitive functioning. DESIGN: Sixteen healthy women with normal menstrual cycles completed the California Verbal Learning Test, a digit span test, and a verbal fluency test in the follicular phase of a normal menstrual cycle and a second time after four monthly injections of the gonadotropin-releasing hormone (GnRH) agonist. Women were randomly assigned to complete a third testing either after resuming cycles in the follicular phase or after three more injections of the GnRH agonist and while wearing an estradiol patch. The control group consisted of 10 women who were tested three times in the follicular phase of their menstrual cycles. RESULTS: Results showed no change in cognitive functioning across sessions or groups in women with suppressed ovarian function. Women who had the highest levels of menopausal symptoms when taking the GnRH agonist did not have significantly lower cognitive functioning. CONCLUSIONS: This study did not find any effect of suppression in ovarian hormones on cognitive performance of young women. PMID- 12082359 TI - Prevention of bone loss and hypoestrogenic symptoms by estrogen and interrupted progestogen add-back in long-term GnRH-agonist down-regulated patients with endometriosis and premenstrual syndrome. AB - OBJECTIVE: To examine the utility of a low-dose estrogen and pulsed progestogen hormone replacement therapy (HRT) regimen for add-back during long-term gonadotropin-releasing hormone-agonist (GnRH-agonist) therapy. DESIGN: A pilot clinical trial conducted at a tertiary referral, academic, reproductive sciences center. The study included 15 patients with endometriosis and 5 patients with severe premenstrual syndrome (PMS). Patients with endometriosis received leuprolide acetate depot 3.75 mg IM monthly until their symptoms had resolved (2 3 months), at which time HRT was initiated along with the GnRH-agonist. Patients with severe PMS received the same treatment with the addition of HRT after 1 month. The HRT regimen consisted of 1 mg oral micronized estradiol daily and 0.35 mg norethindrone daily for 2 days alternating with 2 days without norethindrone. The main outcome measure included bone density assessment in the lumbar spine and femoral neck by dual-energy x-ray absorptiometry at 6- to 12-month intervals. The mean follow-up duration +/- SD while on GnRH-agonist treatment was 31.2 +/- 17 months (for endometriosis patients) and 37.7 +/- 8.4 months (for patients with severe PMS). RESULTS: Bone mineral density was stable after initiation of HRT for the entire follow-up period. No patient had return of pelvic pain or resumption of mood swings after HRT add-back. After the first 3 months of HRT, all women remained amenorrheic. CONCLUSIONS: Long-term GnRH-agonist down-regulation is safe and effective when combined with HRT add-back. Furthermore, on the basis of this small study, the low-dose pulsed progestogen, continuous estrogen HRT regimen seems to be safe for use as add-back therapy in terms of bone health. PMID- 12082360 TI - Comparative effects of tibolone and conjugated equine estrogens with and without medroxyprogesterone acetate on the reproductive tract of female cynomolgus monkeys. AB - OBJECTIVE: To measure the effects of 2 years' treatment with tibolone on the reproductive tract of female monkeys (Macaca fascicularis) in comparison with conventional hormone replacement therapy. DESIGN: Ovariectomized adult female monkeys were randomized for 2 years of treatment into five groups: controls (n = 31); tibolone at 0.05 mg/kg (LoTIB group; n = 30); tibolone at 0.2 mg/kg (HiTIB group; n = 31); conjugated equine estrogens (CEE) at 0.042 mg/kg (CEE group; n = 28); or CEE + medroxyprogesterone acetate (MPA) at 0.167 mg/kg (CEE + MPA group; n = 29). Endpoints included vaginal cytology; uterine weight; histopathologic evaluation of the uterus, vagina, and cervix; histomorphometry of the endometrium; and immunohistochemical detection of the proliferation marker Ki67 and progesterone receptor in endometrial tissue. RESULTS: Endometrial atrophy was found in 29 of 30 and 23 of 31 animals in the LoTIB and HiTIB groups, respectively, compared with 0 of 28 and 11 of 29 in the CEE and CEE + MPA groups, respectively. All ovariectomized control animals had atrophic endometria. No complex or atypical hyperplasia was seen. Simple endometrial hyperplasia of a significant degree was seen in 3 of 31 HiTIB-treated animals, 1 of 30 LoTIB treated animals, 26 of 28 CEE-treated animals, and 16 of 29 CEE + MPA-treated animals, and in none of the control animals. Marked simple endometrial hyperplasia and Ki-67 expression was induced by CEE and partially antagonized by MPA. LoTIB and HiTIB slightly increased endometrial thickness, whereas CEE and CEE + MPA induced a marked increase of 350% and 200%, respectively. Neither LoTIB nor HiTIB increased endometrial proliferation (Ki67 labeling) or induced vaginal keratinization. Endometrial bleeding was not seen in tibolone-treated animals but was present in 10 of 29 animals given CEE + MPA. CONCLUSIONS: The effect of tibolone on the uterus and lower reproductive tract was minimal. The lack of a proliferative response of the endometrium to tibolone, coupled with the lower incidence of endometrial bleeding, suggests that tibolone may have advantages over CEE and CEE + MPA regarding endometrial safety and efficacy. PMID- 12082361 TI - Comparison of physical and emotional side effects of progesterone or medroxyprogesterone in early postmenopausal women. AB - OBJECTIVE: To compare the mood and somatic effects during the initial 2 months of medroxyprogesterone acetate (MPA) or progesterone combined with conjugated equine estrogen (CEE) in early postmenopausal women. DESIGN: Twenty-three nondepressed, early postmenopausal women (average age, 52.5 years) completed a 91-day, single blind pilot study with the following sequence of treatments: 1 week of no substance; 2 weeks of placebo; 2 weeks of progestogen only; 1 week of placebo; and 2 months of "standard hormone replacement therapy cycles," which consisted of (in order) 2 weeks of 0.625 mg CEE, 2 weeks of CEE plus progestogen, 2 weeks of CEE, and 2 weeks of CEE plus progestogen. Ten women who completed the study received MPA (5 mg/day) as their progestogen, and 13 who completed the study received micronized, oil-suspended progesterone (200 mg/day) as their progestogen. All participants made daily assessments of mood using the Profile of Mood States and daily recordings of somatic symptoms. All subjects had plasma follicle-stimulating hormone of greater than 35 IU/L and had not had spontaneous vaginal bleeding for more than 1 year. RESULTS: None of the hormone treatments had a detectable effect on mood. MPA users reported more vaginal bleeding and breast tenderness than progesterone users. CONCLUSIONS: In contrast with the widely held belief among psychiatrists that progesterone depresses mood, neither of the progestogens we used in normal, nondepressed and nonanxious women showed this effect. Absence of an effect on mood was also found when the results of the two progestogens were combined. The lesser side effects of the micronized progesterone-containing regimen suggest that some women may prefer it to an MPA containing regimen. PMID- 12082362 TI - The relationship of race/ethnicity and social class to hormone replacement therapy: results from the Third National Health and Nutrition Examination Survey 1988-1994. AB - OBJECTIVE: To use a nationally representative sample to examine the prevalence of hormone replacement therapy (HRT) use and its relationship to different markers of social class in American women 60 years of age and older. DESIGN: Nationally representative cross-sectional survey with an in-person interview and medical examination. Between 1988 and 1994, 3,479 women aged 60 to 90+ years were examined as part of the National Health and Nutrition Examination Survey III. Mexican Americans, non-Hispanic blacks and much older women were oversampled to produce reliable estimates for these groups. RESULTS: Overall, the number of women who reported ever having used HRT was 37% [confidence interval (CI), 33% 40%] of all women older than 60 years of age; 40% (CI, 37%-41%) of older, non Hispanic white women; 20% (CI, 14%-25%) of non-Hispanic black women; and 24% (CI, 20%-29%) of Mexican American women. HRT was used by 43% (CI, 38%-47%) of women 60 to 70 years old, 37% (CI, 32%-41%) of those 71 to 80 years old, and 20% (CI, 13% 26%) of women older than 80. HRT use was lowest among women who did not complete high school or among those in the lowest family income categories. Among women more than 60 years old who reported having a hysterectomy, 51% (CI, 47%-55%) reported using HRT, whereas only 20% (CI, 17%-23%) of those who had a natural menopause reported using HRT. CONCLUSIONS: Although many women can benefit from HRT, the number of American women who report they have ever used it remains low. More research is needed to examine the implications of racial differences in compliance, patient and physician attitudes toward HRT, and possible environmental barriers that may prevent use. PMID- 12082364 TI - Cyproterone acetate could counteract the benefits of estradiol valerate in oophorectomized cholesterol-fed rabbits. AB - OBJECTIVE: The administration of hormonal replacement treatment to women with an intact uterus needs to be supplemented with progestogenic compounds to avoid endometrial hyperplasia. However, progestins may cancel the beneficial effects of estrogens on the cardiovascular system. The goal of this study was to examine the effects of adding cyproterone acetate to estradiol (E(2))on aorta atherogenesis. DESIGN: Thirty-two cholesterol-fed New Zealand white rabbits were studied for 4 months. The animals underwent laparotomy and were randomly allocated to four groups. Twenty-four rabbits underwent bilateral ovariectomy, and the other eight were sham-operated (group S). The ovariectomized rabbits were allocated to three groups of eight animals each receiving E(2) valerate (group E), E(2) valerate plus cyproterone acetate (group EC), or placebo after sterilization (group C). RESULTS: After 4 months, the cholesterol-rich diet caused atherosclerotic aortic lesions in both treated groups that affected 17.91% +/- 10.19% and 28.16% +/- 7.97% of the aortic surface of groups E and EC, respectively, with a markedly lower aortic plaque size in group E than in groups C and S. Rabbits from group E (but not from group EC) had aortic cholesterol content significantly lower than rabbits from the sham-operated and control groups. CONCLUSION: E(2) valerate reduces aortic atheromatosis in cholesterol-fed, ovariectomized rabbits, and the addition of cyproterone acetate may neutralize this effect. PMID- 12082363 TI - Medroxyprogesterone acetate versus norethisterone: effect on estradiol-induced changes of markers for endothelial function and atherosclerotic plaque characteristics in human female coronary endothelial cell cultures. AB - OBJECTIVE: Progestin addition to estradiol (E(2)) replacement therapy may lead to a deterioration of beneficial effects on the vasculature. The effect of the two clinically most common progestins, medroxyprogesterone acetate (MPA) and norethisterone (NET), during continuous combination with E(2) on the synthesis of markers for coronary endothelial function, atherosclerotic plaque initiation, and plaque formation was investigated in human female vascular cell cultures and compared with that of E(2) alone. DESIGN: Endothelial cell cultures from human female coronary arteries were used to evaluate the effect of progestin addition to E(2) on the production of the following endothelial markers: prostacyclin, endothelin, plasminogen activator inhibitor-1, E-selectin, intercellular adhesion molecule-1, monocyte chemoattractant protein-1 (MCP-1), and the precursor of matrix metalloproteinase-1 (pro-MMP-1). E(2) was tested at 0.1 microM, 1 microM, and 10 microM alone and in equimolar combinations with MPA or NET. The markers were determined by enzyme immunoassays in the cell supernatant. RESULTS: E(2) induced a significant increase of endothelial prostacyclin production and was able to significantly decrease the synthesis of endothelin, plasminogen activator inhibitor-1, E-selectin, and intercellular adhesion molecule-1. Neither MPA nor NET addition negatively interfered with these E(2)-induced benefits. However, MPA antagonized the E(2)-induced significant reduction of MCP-1 synthesis, with the difference between both progestins being significant (p < 0.01). Interestingly, an enhancement of the positive E(2)-effect on pro-MMP-1 production was observed by the addition of both MPA and NET (p < 0.01). CONCLUSION: E(2) can positively influence various markers of endothelial function. Addition of MPA or NET can elicit different effects, which has been demonstrated for the first time in human coronary cell cultures. No impact was found on markers representing primarily vasotonus and thrombogenicity. In terms of MMP-1, which is crucial for atherosclerotic plaque stability, an enhancement of the beneficial E(2) effect was observed. However, regarding MCP-1, contrary effects of progestins cannot be excluded. This indicates that progestins may differ in their effects, particularly in the early stages of atherosclerosis, which has also been supported by other studies. PMID- 12082365 TI - Medroxyprogesterone-induced endocrine alterations after menopause. AB - OBJECTIVE: To evaluate endocrine responses to short-term medroxyprogesterone acetate (MPA) administered as a respiratory stimulant in postmenopausal women with mild nocturnal hypoxemia. DESIGN: Open-label trial with 14-day MPA (60 mg daily) and 3-week follow-up posttreatment. Serum concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol, progesterone, and insulin-like growth factor-I (IGF-I) were assessed at baseline, while on MPA and at the 3-week washout in eight subjects. RESULTS: With MPA, FSH decreased by 47.5% (p < 0.01) and LH by 64.9% (p < 0.001), whereas IGF-I increased by 39.6% (p < 0.01). FSH and LH remained suppressed at the 3-week washout (-23.8%, p < 0.05 and -44.3%, p < 0.01, respectively). IGF-I returned to pretreatment level at the 3-week washout. Neither serum estradiol nor progesterone concentrations changed during or after progestogen therapy. CONCLUSIONS: Daily administration of MPA (60 mg) for 2 weeks has both immediate (FSH, LH and IGF-I) and sustained (FSH, LH) effects up to 3 weeks after treatment. Therefore, prolonged MPA-induced effects should be taken into account when interpreting hormone assessments after progestogen therapy. PMID- 12082366 TI - Formation of 7 alpha-methyl-ethinyl estradiol during treatment with tibolone. AB - OBJECTIVE: To investigate whether tibolone, which is orally used for hormone replacement therapy, is transformed to a derivative of ethinyl estradiol (EE). DESIGN: In 10 young women who received 2.5 mg orally administered tibolone daily between cycle day 19 and 25, blood was obtained before and 0.5, 1, 1.5, 2, 3, 4, 6, 8, and 24 h after the last tablet intake. The concentration of 7 alpha-methyl EE was determined by the gas chromatography with mass spectrometry method. RESULTS: The results demonstrate that, during daily treatment of women with 2.5 mg tibolone, a small proportion of tibolone is transformed to 7 alpha-methyl-EE. The maximal serum concentrations of 125 +/- 40 pg/mL were in the range of the levels of EE observed during treatment with oral contraceptives containing 30 microg EE. CONCLUSIONS: Caution is advisable when considering treatment with tibolone of postmenopausal women with contraindications for estrogens. PMID- 12082367 TI - Should we consider performing oral glucose tolerance tests more frequently in postmenopausal women for optimal screening of impaired glucose tolerance? AB - OBJECTIVE: To investigate an optimal screening protocol for impaired glucose tolerance (IGT) and type II or non-insulin-dependent diabetes mellitus (DM) by using fasting plasma glucose (FPG) and oral glucose tolerance test (OGTT) in postmenopausal women. DESIGN: One hundred consecutive postmenopausal women were screened with FPG determination, and then all underwent an OGTT. Basal serum lipid and insulin levels of these women were also determined. Insulin sensitivity was determined by using the homeostasis model assessment. Receiver operating characteristic analysis was performed to determine the efficacy of these variables in detecting women with IGT and DM, and optimal cutoff values were determined. RESULTS: FPG with a cutoff value of 98 mg/dL had the best combination of sensitivity (71%) and specificity (76%) for the detection of IGT and DM. Combined FPG and body mass index screening (with the optimal cutoff value of 26.5 kg/m2) improved the sensitivity to 96% but decreased the specificity to 47%. This combined screening protocol detected 94% of the women with IGT and all diabetic women. CONCLUSIONS: Given that IGT and DM are common among postmenopausal women and DM can be prevented by nonpharmacologic interventions in women with IGT, OGTT may be used more frequently among these women. Our data indicate that for optimal screening of non-insulin-dependent DM and IGT, OGTT should be considered in postmenopausal women, especially when risk factors in addition to age are present. This model may detect most of the women with IGT and almost all diabetic women. PMID- 12082369 TI - International Labor Office Classification System in the age of imaging: relevant or redundant. AB - The 1980 International Labor Office International Classification of Radiographs of Pneumoconioses is a widely used epidemiologic tool with a storied past. This article reviews its development and examines its applications to occupational lung disease and the controversies generated in that process. The question of its relevancy to current imaging practices is discussed. PMID- 12082370 TI - Occupational and environmental thoracic malignancies. AB - Lung cancer is the most common thoracic malignancy caused by exposures at work and in the environment. The most unique thoracic malignancy is mesothelioma, because it is relatively rare and one of only a few neoplasms for which one specific inciting agent-asbestos-has been identified. Based on epidemiologic studies, approximately 15% of lung cancers in men and 5% of lung cancers in women are caused by occupational exposures. The International Agency for Research on Cancer has devised a rating system by which, based on animal and human data, they assign an agent, mixture, or exposure circumstance to one of five categories, ranging from group 1 (agent is carcinogenic to humans) to group 4 (agent is probably not carcinogenic to humans). Group 1 pulmonary carcinogens reviewed in this article include arsenic, asbestos, beryllium, bis (chloromethyl) ether, cadmium, chromium (IV), mustard gas, nickel, radon, and silica. The clinical presentation and pathology of lung cancers and mesothelioma caused by such exposures do not differ from those of cancers caused by other factors. The key to the recognition of a thoracic malignancy caused by workplace or environmental exposures is clinical suspicion and consideration of all causes for the disease present. Recognition of an exposure-related case of lung cancer or mesothelioma can aid in the identification of excess risk for a whole workforce or community and can lead to actions to reduce exposure, thus preventing future cases. In addition, such recognition allows the individuals struck by devastating illness to exercise their legal rights to compensation if so desired. PMID- 12082371 TI - Imaging of thoracic occupational and environmental malignancies. AB - The imaging features of occupational lung cancer are similar to those of nonoccupational cancer. Occupational lung cancer in patients with asbestos exposure must be differentiated from mimics such as round atelectasis and fissural pleural plaques. Mesothelioma remains a largely incurable tumor, though treatment options are expanding. CT, MRI, and PET scanning may all have complementary roles in staging mesothelioma. PMID- 12082372 TI - Portable chest radiographs in the intensive care units: referral patterns and estimated cumulative radiation exposures. AB - The objective of this article was to determine the frequency of referrals for portable chest radiographs from medical and (noncardiac) surgical intensive care units (NICU and SICU) and their respective stepdown units (NICA and SICA). Additionally, the cumulative entrance skin exposure (ESE) using an ion chamber was determined. We retrospectively reviewed the medical records of all adult patients admitted to the MICU, SICU, MICA, and SICA at a tertiary referral center during a 6-month interval. The duration of stay and the number of portable chest radiographs were determined for each patient. The measured ESEs from all portable radiography units ranged from 5 to15 mR (average: 10mR). The cumulative radiation exposure for each patient was calculated. There were 567 patients admitted to the units: 146 surgical and 421 medical. Their ages ranged from 15 to 87 years. The duration of stay varied from 1 to 68 days. A total of 3,794 portable chest radiographs were obtained. The number of radiographs per patient varied from 1 to 94. The number of radiographs and the corresponding cumulative radiation doses were as follows: 406 patients (72%) had fewer than five radiographs (<50 mR); 76 (13%) had five to 10 radiographs (<100 mR); 35 (6%) had 11 to 20 (<200 mR); and 50 (9%) had more than 20 chest radiographs (>200 mR). The cumulative ESE ranged from 10 to 940 mR. It exceeded 450 mR in only nine (1.5%) patients. Most (73%) patients undergoing intensive care undergo fewer than five radiographs during their stay in the units. Patient exposure from portable chest radiographs in this population is less than the average annual exposure from background radiation in the USA (450-500 mR), and is much less than the average annual exposure from teratogenic radiation. PMID- 12082373 TI - Role of computed tomography in guiding the management of peripheral bronchopleural fistula. AB - The present study was designed to elucidate whether demonstration of a peripheral bronchopleural fistula on CT correlated with the need for surgical management. We retrospectively identified 33 patients, 24 males and nine females, mean age 38 years, with clinical diagnosis of peripheral bronchopleural fistula and whose chest CT scans and medical charts were reviewed. Each chart was reviewed to identify the cause of the peripheral bronchopleural fistula and its treatment. Treatment decisions were categorized as surgical or conservative. Each chest CT was evaluated for the cause of peripheral bronchopleural fistula as follows: bulla(e), lung abscess/necrotizing pneumonia, neoplasms, peripheral bronchiectasis, and trauma. The peripheral bronchopleural fistula was classified as visible on CT if a distinct channel between the lung or a peripheral bronchus and the pleura was seen on the lung windows. We found that CT was useful in guiding surgery by identifying and localizing the cause of the peripheral bronchopleural fistula in the 55% (18/33) of patients who required surgery. The peripheral bronchopleural fistula or its probable cause was identified in 91% (30/33) as follows: bulla(e) (n = 12), lung abscess/necrotizing pneumonia (n = 11), peripheral bronchiectasis (n = 5), malignancy (n = 1), and posttraumatic pneumatocele (n = 1). The peripheral bronchopleural fistula was right-sided in 24, left-sided in nine, and was visible on CT in 36% (12/33). Among the patients with bullae, 58% (7/12) required surgery; however, the peripheral bronchopleural fistula was visible on CT in only 8% (1/12). Among the 21 patients without bulla(e), the peripheral bronchopleural fistula was visible on CT in 52% (11/21). When the fistula was visible in this subgroup, 73% (8/11) required surgery compared with 30% (3/10) in whom the fistula was not visible (p = NS; Fisher exact). In conclusion, CT was useful in guiding surgery by identifying and localizing the peripheral bronchopleural fistula or its probable cause. Peripheral bronchopleural fistulas caused by bulla(e) were less likely to be visible on CT (p < 0.05). Excluding patients with bulla(e), our data suggest a trend toward the need for surgical management for patients in whom the peripheral bronchopleural fistula was visible on CT. PMID- 12082374 TI - Azygoesophageal recess. AB - Knowledge of the normal radiologic appearance and manifestations of disease in the azygoesophageal recess can facilitate the detection and diagnosis of intrathoracic disease. The azygoesophageal recess is located lateral or posterior to the esophagus and anterior to the spine. It extends from the anterior turn of the azygous vein to the aortic hiatus. The configuration of the azygoesophageal recess is normally a smooth arc convex to the left. Because the right lung extends into the recess, pathology in this region can often be detected radiographically and typically manifests as increased opacity and/or as a contour abnormality. In adults, convexity directed to the right and immediately below the level of the azygous vein on a frontal radiograph is abnormal and requires further evaluation. PMID- 12082375 TI - Minute pulmonary meningothelial-like nodules: high-resolution computed tomography and pathologic correlations. AB - Minute pulmonary meningothelial-like nodules (MPMNs), previously known as minute pulmonary chemodectomas, are relatively rare lesions. They are small (1-3 mm) and often multiple. Pathologically, they represent an interstitial nodular proliferation of small oval or spindle-shape cells arranged in a "zellenballen" nesting pattern. The function and origin of the cells are unknown. These nodules are associated with specific conditions, including thromboembolism, cardiac disease, and malignancy. We describe a patient with MPMNs and adenocarcinoma of the lung in whom HRCT showed tiny (1-3 mm in diameter) nodules of ground-glass attenuation. PMID- 12082376 TI - Radiographic findings in ovarian hyperstimulation syndrome. AB - The authors report a patient with ovarian hyperstimulation syndrome who had findings on chest radiographic. Bilateral pleural effusions and bibasilar partial atelectasis were the preeminent features. The pleural effusions resulted from accompanying ascites. Ovarian hyperstimulation syndrome occurs in the postovulatory or post-oocyte retrieval phase, 5 to 7 days after administration of human chorionic gonadotropin. Awareness of imaging findings should facilitate the diagnosis of this syndrome. PMID- 12082377 TI - Refractory chylothorax in hepatic cirrhosis: successful treatment by transjugular intrahepatic portosystemic shunt. AB - Patients with cirrhosis and portal hypertension have increased thoracic duct lymph flow. Correction of portal hypertension is associated with decreases in thoracic duct flow. The authors present a case of rapid resolution of refractory chylothorax caused by thoracic duct injury proven by lymphangiography and helical CT scan in a patient with cirrhosis of the liver by using a transjugular intrahepatic portosystemic shunt to decrease portal pressure and thereby reduce thoracic duct lymph flow. PMID- 12082378 TI - Proceedings of the Festschrift for Professor John Walker-Smith. London, United Kingdom, September 29, 2000. PMID- 12082379 TI - Toward a molecular understanding of complex childhood enteropathies. PMID- 12082380 TI - Studies of the small intestine in persistent diarrhea and malnutrition: the Gambian experience. PMID- 12082381 TI - The gut-brain axis in childhood developmental disorders. PMID- 12082382 TI - The enteropathy of celiac disease. PMID- 12082383 TI - Feeding genes. PMID- 12082384 TI - The biochemical basis of immune enteropathy. PMID- 12082385 TI - The cause of Th2 to Th1 imbalance in asthma: a function of exposure to typhoid and tuberculosis? PMID- 12082386 TI - Development of the intestinal mucosal barrier. PMID- 12082387 TI - The pediatric esophagus comes of age. PMID- 12082388 TI - Getting to grips with gastric pathology. PMID- 12082389 TI - Transplantation--new beginnings and new horizons. PMID- 12082390 TI - Role of food antigen elimination in treating children with persistent diarrhea and malnutrition in Zambia. PMID- 12082391 TI - Enteropathogenetic Escherichia coli: from inpatient to intimin. PMID- 12082392 TI - Import and export of pediatric gastroenterology. PMID- 12082393 TI - Oral rehydration: an evolving solution. PMID- 12082394 TI - Recollections and reflections on the medical life, student and doctor, 1954 to 2000. PMID- 12082395 TI - A brief reflection on the development of human retrovirology: the past, the present and the future. PMID- 12082396 TI - Functional analysis of Kaposi's sarcoma-associated herpesvirus RTA in an RTA depressed cell line. AB - OBJECTIVE: The viral transcriptional activator encoded by Kaposi's sarcoma associated herpesvirus (KSHV) open reading frame 50 (RTA) is expressed in the immediate-early phase of reactivation. We isolated an RTA-depressed cell line, BLS50-4, by subcloning from the KSHV-infected cell line, BCBL-1. RESULTS: In addition to RTA, induction of some lytic gene expressions was also remarkably reduced in BLS50-4 cells, but that of the K8.1 and ORF65 gene expressions was not. Both the replication of the KSHV genome and the release of KSHV DNA into the medium were greatly reduced. Transfection of RTA into BLS50-4 cells restored the expression of K9 (vIRF) and ORF59, but not K8.1. CONCLUSIONS: Thus, we showed that expression of late genes was not directly controlled by RTA, unlike the other groups concluded. We also showed that, by isolating the RTA-depressed cell line, the RTA protein had a critical role in viral DNA replication and the expression of several lytic genes. PMID- 12082397 TI - Enhancement of cellular and humoral immune responses to human immunodeficiency virus type 1 Gag and Pol by a G/P-92 fusion protein expressing highly immunogenic Gag p17/p24 and Pol p51 antigens. AB - OBJECTIVES: Immunity to the human immunodeficiency virus type-1 (HIV-1) G/P-92 fusion protein consisting of highly immunogenic regions of Gag (p17 and p24) and Pol (p51) expressed in recombinant vaccinia virus (vG/P-92) was compared with responses to the entire viral Gag-Pol precursor protein (vVK1). STUDY DESIGN/METHODS: We analyzed the level of Gag and Pol protein expression in vG/P 92-infected cells as well as the ability of the G/P-92 fusion protein to form virus-like particles (VLP) in infected cultures. The efficacy of vG/P-92 and vVK1 vaccines was evaluated in a murine model by measuring T helper (Th), cytotoxic T lymphocyte (CTL), and antibody responses to Gag and Pol antigens. RESULTS: The deletion of a frameshift site resulted in an increased level of Pol in cells expressing the G/P-92 fusion protein. Particles budding from the plasma membrane were detected in both vG/P-92- and vVK1-infected cells, but the release of VLP was less efficient from cells expressing the G/P-92 fusion protein than the entire gag-pol gene product. Immunization with vG/P-92 vector elicited a higher level of cellular and humoral responses to both Gag and Pol antigens than the vVK1 vaccine. CONCLUSIONS: The enhanced immunogenicity of the G/P-92 fusion protein compared with the entire viral gag-pol gene product might be related to a higher intracellular level of Pol and Gag expression due to the deletion of a frameshift site and less efficient transport of VLP from vG/P-92-infected cells, respectively. PMID- 12082398 TI - In vivo emergence of drug-resistant mutations at less than 50 HIV-1 RNA copies/mL that are maintained at viral rebound in longitudinal plasma samples from human immunodeficiency virus type-1-infected patients on highly active antiretroviral therapy. AB - OBJECTIVE: Emergence of human immunodeficiency virus type-1 (HIV-1) genotypic drug resistance is generally attributed to noncompliance, poorly absorbed drugs, or drug-to-drug interaction. Attempts to determine emerging genotypic drug resistance from study subjects on highly active antiretroviral therapy (HAART) relied on insensitive polymerase chain reaction (PCR) techniques, revealing wild type HIV-1 or precursor resistant genotypes from few plasma samples successfully amplified with <50 copies/mL. STUDY DESIGN/METHODS: In this analysis, using Applied Biosystems' ViroSeq HIV-1 Genotyping Systems, Version 2.0 (Applied Biosystems, Foster City, CA, USA) and the supplemental, for research use only, nested PCR primers, genotypic drug resistance was determined in longitudinal plasma samples from 11 study subjects on HAART. RESULTS: In 4 of 11 study subjects, newly emerging genotypic primary resistant mutations were detected in plasma samples with <50 copies/mL. Most of these primary drug-resistant mutations were detected in subsequent longitudinal samples with detectable viral load (viral breakthrough). CONCLUSIONS: This analysis suggests sufficient viral replication <50 copies/mL to generate genotypic drug resistance in study subjects on suppressive HAART. PMID- 12082399 TI - Human papillomavirus DNA is present in a subset of unselected breast cancers. AB - OBJECTIVE: The major molecular events in the genesis of most breast cancers are unknown. However, human papillomaviruses (HPV) have been reported to be found in a significant portion of breast cancers of women with concomitant cervical intraepithelial neoplasia III. To investigate a potential HPV-breast cancer link, we carried out a small survey to identify HPV in unselected, general breast cancer tissues. STUDY DESIGN/METHODS: Deoxyribonucleic acid (DNA) was isolated from 17 breast cancer tissues (and one cervical swab) taken from our local, randomly selected patient population. Two different previously characterized broad-spectrum primer sets (targeting the E6/E7 or L1 regions) were used to amplify HPV DNA, and another primer set was used to amplify the ColE1/pBR322 origin of replication by polymerase chain reaction amplification. The polymerase chain reaction product DNA was analyzed by dot blot hybridization with HPV-16, 18, -31, or pRB322 DNA probes. Total cellular DNA was also analyzed by one- and two-dimensional Southern blot analysis. Finally, the E6/E7 polymerase chain reaction products were cloned, sequenced, and compared to previously cloned HPV types. RESULTS: Polymerase chain reaction/dot blot analysis by both the HPV E6-E7 and L1 primer sets identified the same 6 out of 17 (35%) breast cancers as being HPV positive. ColE1/pBR322 origin targeted polymerase chain reaction/dot blot analysis failed to identify plasmid contamination. One- and two-dimensional Southern blot analysis showed that the breast cancers specimens contained significant levels of HPV DNA and that the viral DNA was largely episomal. The sequences of the HPV clones demonstrated that HPV-16, -18, and possibly type 11 were present within the breast cancer specimens. Furthermore, the HPV sequences cloned from the cervical swab and breast cancer of the same patient were found to be identical. CONCLUSIONS: These data suggest that HPV may be associated with a significant subset of breast cancers, and further suggest that additional studies are warranted. PMID- 12082400 TI - Chronic stavudine exposure induces hepatic mitochondrial toxicity in adult Erythrocebus patas monkeys. AB - OBJECTIVES: To understand the mitochondrial mechanisms underlying the lactic acidosis and hepatic steatosis seen in some HIV-1-infected individuals after long term stavudine (d4T) exposure, we have explored mitochondrial integrity in adult monkeys (Erythrocebus patas) given a daily human equivalent dose of d4T for 78 days. STUDY DESIGN/METHODS: Three Erythrocebus patas (patas) monkeys were given 3 mg d4T orally twice daily (total 6 mg d4T), or approximately 1.2 mg d4T/kg body weight per day, for 78 days and compared with 3 unexposed animals. Blood taken from controls and from treated monkeys before and after drug exposure was subjected to a complete clinical chemistry profile. Liver and skeletal muscles were examined for oxidative phosphorylation enzyme specific activities, mitochondrial deoxyribonucleic acid (mtDNA) quantity by slot blot, and mtDNA integrity by Southern blot. RESULTS: Clinical chemistry assays demonstrated few significant differences; however, one d4T-exposed monkey had a serum lactate of 8.1 mmol/L after 78 days of oral d4T ingestion. Specific activities of oxidative phosphorylation Complexes I, II, and IV were significantly altered in both livers and skeletal muscles from the d4T-exposed animals, compared with the controls (p < or = 0.05). Significant depletion of mitochondrial DNA was observed in livers of drug-exposed monkeys, but not in skeletal muscle (p < or = 0.05). Further examination of liver DNA by Southern blot confirmed hepatic mtDNA depletion in drug exposed animals. CONCLUSIONS: The data suggest that direct examination of the liver may be required to elucidate clinical d4T-induced hepatotoxicity related to mitochondrial damage. PMID- 12082443 TI - [The lung specialist and obesity]. PMID- 12082401 TI - Cervicovaginal shedding of TT virus in HIV-infected women. AB - OBJECTIVE: TT virus (TTV) is frequently detected in the serum and in other body fluids of humans. Recently TTV-specific deoxyribonucleic acid has been detected in cervical specimens from apparently healthy women and in seminal fluid, suggesting that sexual transmission may be common. STUDY DESIGN/METHODS: TT virus deoxyribonucleic acid prevalence was assessed in paired samples of blood and cervical smears from 110 human immunodeficiency virus-positive women. Detection and typing of human papillomavirus (HPV) present in cervical smears was also performed. RESULTS: The prevalence of TTV-deoxyribonucleic acid in cervical smears was 16.4%, without significant difference (p = 0.81) between HPV-positive (18.6%) and -negative (14.9%) samples. The distribution of high/middle and low risk HPV types was similar in TTV-positive and -negative samples. On the contrary, women with multiple HPV infections had a significantly higher TTV deoxyribonucleic acid prevalence (60.0%) than HPV-negative women (p = 0.04). CONCLUSIONS: TT virus excretion in the female genital tract of human immunodeficiency virus-infected women is common, further supporting sexual transmission of this virus. PMID- 12082444 TI - [Obesity: epidemiology, pathophysiology and extra-respiratory complications]. AB - Obesity is the most common nutritional disease in industrialized countries and constitutes an important public health issue. Strictly speaking, obesity is an excess of adipose tissue. In practice, body mass index (BMI)=weight (kg)/height (m)(2) is used to determine the degree of excess body weight in a patient. A BMI >=30 kg/m(2), which is associated with significantly higher morbidity and mortality, is generally used as a threshold value for obesity. Despite this measurement, obesity includes a very wide range of variable symptoms. Development of obesity is a multifactorial phenomenon implicating biological, genetic, behavioral, and environmental factors whose interaction and inter-association vary from individual to individual. There are many extra-respiratory complications of obesity which may be somatic (cardiovascular, mechanic, metabolic.), psychological, or social. Abdominal adipose tissue plays an important role in metabolic and vascular complications of obesity independently of the degree of corpulence. As an introduction, we discuss here the definitions concerned as well as the epidemiology, basic pathophysiological mechanisms, and extra-respiratory complications of adult obesity. PMID- 12082445 TI - [Respiratory function in the obese subject]. PMID- 12082446 TI - [Alveolar hypoventilation in the obese: the obesity-hypoventilation syndrome]. AB - The obesity-hypoventilation syndrome (or alveolar hypoventilation in the obese) is a new name for an old syndrome, Pickwickian syndrome. It is defined as chronic alveolar hypoventilation (PaO(2)<70 mmHg, PaCO(2) > 45 mmHg) in obese patient with a body mass index > 30 kg/m(2) who have no other respiratory disease explaining the gas anomalies. The large majority of obese subjects are not hypercapnic, even in case of severe obesity. There are three principal causes explaining alveolar hypoventilation in obese subjects: high cost of the work of respiration, dysfunction of the respiratory centers, repeated episodes of nocturnal obstructive apnea. The obesity-hypoventilation syndrome is generally found in males aged over 50 years. Exercise-induced breathlessness is a constant finding. Diagnosis is often made after an episode of severe respiratory failure. Associated diseases favored by obesity are frequent: diabetes, high blood pressure, heart disease. By definition, there is a hypoxemia-hypercapnia syndrome persisting after an acute episode. Spirography usually demonstrates moderate volume restriction. Pulmonary hypertension is frequent but not constant. Obesity hypoventilation syndrome must be distinguished from obstructive sleep apnea, although the two conditions are often associated. Obstructive sleep apnea may be absent in certain patients with obesity-hypoventilation syndrome (we have had several cases) and inversely, obesity is not observed in certain patients with obstructive apnea. It should be recalled that the term Pickwickian syndrome designates obesity-hypoventilation syndrome (with or without obstructive apnea) and not obstructive sleep apnea syndrome. PMID- 12082447 TI - [Sleep apnea syndrome and obesity]. AB - Obesity is a main risk factor for sleep apnea syndrome (SAS). The prevalence of SAS is especially high in massive obesity and in visceral obesity. The mechanisms of obstructive apneas in obesity are poorly known, but an increase in upper airway collapsibility probably plays an important role. Several cardiorespiratory complications of SAS, especially systemic arterial hypertension, diurnal alveolar hypoventilation and pulmonary arterial hypertension, are more frequent and more severe in obese patients. An important weight loss resulting from surgical treatment of obesity is often associated with a dramatic decrease in apnea hypopnea index in patients with massive obesity. In patients with moderate obesity, dietary weight loss is associated with varying degrees of improvement in SAS. Pharyngoplasty and anterior mandibular positioning devices have a low success rate in patients with massive obesity. Nasal continuous positive airway pressure is often the only effective treatment in obese SAS patients. PMID- 12082448 TI - [Exercise tests in obese subjects]. PMID- 12082449 TI - [Pulmonary hypertension and obesity]. AB - Obesity is a morbid condition with hemodynamic consequences affecting the systemic and pulmonary circulations leading to a risk of pulmonary hypertension. Data in the literature do not argue in favor of a direct relationship between pulmonary hypertension and obesity. These two conditions appear to be two distinct entities, the different co-morbidities observed in obesity favoring pulmonary hypertension. Certain co-morbidities, for instance use of anorexic agents, exhibit a clear relationship with pulmonary hypertension. There is also a possible relationship with left ventricular failure, hypoxemia, and other respiratory disorders (including obstructive sleep apnea), hypothyroidism, and thomboembolism. PMID- 12082450 TI - [Acute respiratory failure in obesity]. AB - Obstructive sleep apnea, obesity-related hypoventilation - a hypoventilation which is independent of apneas and increased by sleep -, and hypoxemia related to local ventilation-perfusion disorders are the main mechanisms of respiratory failure occurring during acute respiratory decompensation following an often minimal triggering event. Non-invasive ventilation has been found to be an effective treatment, particularly with a ventilator capable of maintaining positive expiratory and pressure. The level of the expiratory positive airway pressure must be adapted to cure episodes of obstructive apnea or hypopnea. The level of the inspiratory positive airway pressure (pressure support ventilator), or the tidal volume (volume-controlled ventilator) must be adapted to correct the residual hypoventilation. These adaptations can be made by proper assessment of nocturnal SaO(2) recordings. In particularly severe cases, use of endotracheal ventilation may be necessary to control a state of shock or consciousness disorders incompatible with the patient cooperation necessary for non-invasive ventilation. PMID- 12082451 TI - [Anesthesia and obesity]. AB - Problems encountered during anesthesia procedures in obese subjects is related to the level of overweight. Obesity multiplies the effect of general anesthesia on the respiratory function and increases the postoperative risk of cardiovascular disorders and deep vein thrombosis. The pharmacokinetic behavior of most general anesthesia drugs is affected by the mass of adipose tissue producing a prolonged less predictable effect. Control of airway permeability and continence is also a major problem with predictable or unpredictable difficulties with endotracheal intubation. Locoregional anesthesia, which avoids the intubation problem, is difficult to implement and does not provide satisfactory results in all cases. The preoperative work-up should assess the consequences of obesity, particularly concerning the respiratory, cardiovascular, and metabolic systems. A proper work up allows the anesthesist to provide the obese subject with well-informed information on the risk and benefit of proposed options. PMID- 12082452 TI - [Treatment of obesity]. AB - There are three main goals to the treatment of obesity: prevention and treatment of the complications, realistic reduction of excess weight, psychological well being. Each individual is in a specific clinical situation and interindividual variability is wide so stereotypic therapeutic schemes are of little use. Treatment calls upon physical exercise, dietary counseling, drugs, and exceptionally surgery. Psychological support is an integral part of patient management. Treatment of respiratory complications focuses on preventing thromboembolism and management of sleep apnea syndrome and alveolar hypoventilation. PMID- 12082453 TI - Phylogenetic analysis of intestinal microflora indicates a novel Mycoplasma phylotype in farmed and wild salmon. AB - All studies of the microbial community of the gastrointestinal tract of salmon to date have employed culture-based approaches, typically on pond- or tank-raised, freshwater animals. We present a phylogenetic survey of the bacterial populations present in the distal intestine of salmon from three different marine locations in Europe. This was accomplished through PCR amplification, cloning, and sequencing of partial 16S rDNA genes from microbial community DNA isolated from the contents of the GI tract distal to the pyloric ceca. Using this approach, the intestinal microbial communities of wild salmon from Scotland and pen-raised salmon from Scotland and Norway were compared. The predominating bacterial populations detected were Acinetobacter junii and a novel Mycoplasma phylotype. This Mycoplasma phylotype apparently comprised approximately 96% of the total microbes in the distal intestine of wild salmon. Substantial differences in intestinal microbial community composition and diversity were observed between the two groups of pen-raised salmon, which, in addition to geographical separation, were raised on different feeds. The microbial profiles found in this study were substantially different from those indicated in earlier culture-based studies for several species of fish, presumably because of the culture independent techniques employed. Further, analysis of short-chain fatty acids in the digestive tract indicated that the decreasing redox gradient from proximal to distal reaches common to homeothermic animals was absent in salmon, and that the bacterial fermentation levels were much lower than are reported in homeothermic animals. PMID- 12082454 TI - Distribution of capsulated bacterioplankton in the North Atlantic and North Sea. AB - In laboratory experiments, bacterioplankton were incubated under different nutrient conditions, and the percentage of bacteria exhibiting a polysaccharidic capsule (capsulated bacteria) and that of CTC (cyanotetrazolium chloride) positive and therefore metabolically highly active bacteria were determined. In these seawater cultures amended with nutrients more than 95% of the CTC-positive cells exhibited a capsule. During two cruises, one to the North Atlantic and one to the North Sea, we investigated the distribution of capsulated bacteria throughout the water column. Capsulated bacteria were generally more abundant in eutrophic surface waters than in deeper layers or more oligotrophic regions. In the upper 100 m of the North Atlantic, about 6-14% of the total bacterioplankton community was capsulated, while in the layers below 100 m depth, 97% of the bacteria lacked a visible capsule. The percentage of capsulated bacteria correlated with bacterial abundance and production, and chlorophyll a concentration. Also, the bioavailability of DOC (dissolved organic carbon), estimated by the ratio between bacterial production and DOC concentration, significantly correlated with the percentage of capsulated bacteria. In the North Sea, the contribution of capsulated bacteria to the total number of bacteria decreased from the surface (3 m depth) to the near-bottom (25-35 m) layers from 20% to 14% capsulated bacteria. In the nearshore area of the North Sea, about 27% of the bacteria exhibited a capsule. Overall, a pronounced decrease in the contribution of capsulated bacteria to the total bacterial abundance was detectable from the eutrophic coastal environment to the open North Atlantic. Using this epifluorescence-based technique to enumerate capsulated bacterioplankton thus allowed us to routinely assess the number of capsulated bacteria even in the oceanic water column. Based on the data obtained in this study we conclude that almost all metabolically highly active bacteria exhibit a capsule, but also some of the metabolically less active cells express a polysaccharide capsule detectable with this method. PMID- 12082455 TI - A phase I/II trial of rAd/p53 (SCH 58500) gene replacement in recurrent ovarian cancer. AB - PURPOSE: To determine the safety, gene transfer, host immune response, and pharmacokinetics of a replication-deficient adenovirus encoding human, recombinant, wild-type p53 (SCH 58500) delivered into the peritoneal cavity (i.p.) alone and sequentially in combination with platinum-based chemotherapy, of patients with recurrent ovarian, primary peritoneal, or fallopian tube cancer containing aberrant or mutant p53. METHODS: SCH 58500 was administered i.p. to three groups of patients with heavily pretreated recurrent disease. Group 1 (n=17) received a single dose of SCH 58500 escalated from 7.5 x 10(10) to 7.5 x 10(12) particles. Group 2 (n=9) received two or three doses of SCH 58500 given alone for one cycle, and then with chemotherapy for two cycles. The SCH 58500 dose was further escalated to 2.5 x 10(13) particles/dose in group 2. A third group (n=15) received a 5-day regimen of SCH 58500 given at 7.5 x 10(13) particles/dose per day i.p. alone for cycle 1 and then with intravenous carboplatin/paclitaxel chemotherapy for cycles 2 and 3. RESULTS: No dose-limiting toxicity resulted from the delivery of 236/287 (82.2%) planned doses of SCH 58500. Fever, hypotension abdominal complaints, nausea, and vomiting were the most common adverse events. Vector-specific transgene expression in tumor was documented by RT-PCR in cells from both ascitic fluid and tissue biopsies. Despite marked increases in serum adenoviral antibody titers, transgene expression was measurable in 17 of 20 samples obtained after two or three cycles of SCH 58500. Vector was detectable in peritoneal fluid by 24 hours and persisted for as long as 7 days whereas none was detected in urine or stool. There was poor correlation between CT scans and CA125 responses. CA125 responses, defined as a greater than 50% decrement in serum CA125 from baseline, were documented in 8 of 16 women who completed three cycles of the multidose regimen. CONCLUSION: CT scans are not a valid measure of response to i.p. SCH 58500 due to extensive adenoviral-induced inflammatory changes. Intraperitoneal SCH 58500 is safe, well tolerated, and combined with platinum-based chemotherapy can be associated with a significant reduction of serum CA125 in heavily pretreated patients with recurrent ovarian, primary peritoneal, or fallopian tube cancer. PMID- 12082456 TI - Long term follow-up of patients with recurrent ovarian cancer after Ad p53 gene replacement with SCH 58500. AB - OBJECTIVE: We have previously reported the safety, efficient gene transfer, and favorable CA125 responses of individuals with recurrent ovarian cancer treated by p53 gene replacement with the adenoviral vector SCH 58500. The purpose of the present investigation was to evaluate the long-term follow-up of these heavily pretreated patients subsequent to SCH 58500 dosing. METHODS: Patients (n=36) were treated with either single-dose SCH 58500 in the phase I study or with multiple doses (MD) of SCH 58500 over multiple cycles in combination of platinum-based chemotherapy in the phase I/II portion of the study. Five patients were initially treated in the single-dose group and re-enrolled in the MD group. The MD group was evaluated both without the re-enrolled patients as MD1 (n=19), and as MD2 (n=24), which included them. Patients who were only treated on the single-dose arm were designated as SD (n=12). Most patients received additional chemotherapy at the discretion of their physicians on completion of the trial. The current analysis is a retrospective sequential cohort survival analysis. RESULTS: The first patient was treated in March 1997 and the last patient completed SCH 58500 in September 1998. There was no difference in age at diagnosis, Karnofsky performance status, interval between diagnosis to SCH 58500, prior cycles or regimen of chemotherapy, platinum-free interval, percent platinum refractory patients, pretreatment CA125, or largest tumor volume between groups. Both MD groups had a slightly longer chemotherapy-free interval before SCH 58500 than the SD group. Median survival of individuals who received MD SCH 58500 with chemotherapy was 12-13.0 months, compared to only 5 months for those treated with SD SCH 58500. There are 10 long-term survivors more than 20 months after MD treatment for recurrent disease compared to only 2 long-term survivors after SD SCH 58500. CONCLUSION: The 12- to 13.0-month median survival in a heavily pretreated population with recurrent ovarian cancer compares favorably to the 16 month median survival for individuals treated with paclitaxel at the time of initial recurrence of this disease and is more than double the 5-month survival seen with palliative radiotherapy or paclitaxel failure. These data suggest that further study of SCH58500 is clearly indicated. PMID- 12082457 TI - Selective killing of cancer cells based on translational control of a suicide gene. AB - The translation initiation factor, eIF4E, is commonly overexpressed in solid tumors. This elevation allows for efficient translation of mRNA that are normally repressed by their 5' untranslated region, many of which encode growth-promoting proteins. This property was exploited to modulate the synthesis of HTK at the translational level to selectively kill cancer cells. Various breast cancer cell lines can efficiently synthesize HTK from the translationally regulated mRNA, whereas normal cells cannot. Accordingly, only cancer cells were killed at low concentrations of ganciclovir. By altering the expression of eIF4E, it was possible to modulate the sensitivity of various cell lines to ganciclovir. PMID- 12082458 TI - Modulation of the atypical multidrug-resistant phenotype by a hammerhead ribozyme directed against the ABC transporter BCRP/MXR/ABCG2. AB - The phenomenon of multidrug resistance (MDR) in human cancers is one of the major causes of failure of chemotherapy. A recently identified new member of the superfamily of ATP-binding cassette transporters, breast cancer resistance protein (BCRP), was demonstrated to confer an atypical multidrug-resistant phenotype to tumor cells. To overcome the BCRP-mediated drug resistance, a specific anti-BCRP hammerhead ribozyme was introduced into the human gastric carcinoma cell line, EPG85-257RNOV, exhibiting an atypical MDR phenotype. By this approach, the expression levels of the targeted BCRP-encoding mRNA and the BCRP transport protein were decreased to the low constitutive expression level that was observed in highly drug-sensitive parental gastric carcinoma cells. In addition, in the anti-BCRP ribozyme-treated cells, the cellular drug accumulation was dramatically increased to the level measured in drug-sensitive cells. These effects were accompanied by an extensive reversal of the drug-resistant phenotype of more than 80%. Because additional mechanisms contribute to the multimodal mediated MDR phenotype exhibited by this gastric carcinoma cell line, the data suggest that the BCRP-mediated contingent to the drug resistance was overcome nearly completely. Moreover, the data indicate that ribozyme-based gene therapy may be clinically applicable in preventing and reversing BCRP-mediated atypical MDR. PMID- 12082459 TI - Inhibition of angiogenesis in vitro by alphav integrin-directed antisense oligonucleotides. AB - The integrin alpha v beta 3 plays a central role in angiogenesis. In this study, we used antisense oligodeoxyribonucleotides (ONs) directed against the alpha v subunit of alpha v beta 3 to inhibit integrin expression. Ten ON sequences, which were selected by systematic alignment of computer-predicted secondary structures of alpha v mRNA, were transfected into human umbilical vein endothelial cells (HUVECs). Following stimulation by PMA, five antisense ONs significantly inhibited alpha v mRNA and protein expression in activated HUVEC at a concentration of 0.05 mciroM with complete prevention of PMA-induced alpha v up regulation by the most potent antisense ON. Inhibition of alpha v expression was associated with significant inhibition of migration of HUVEC by 28% and had no effect on proliferation and apoptosis. Moreover, transfection of antisense ON inhibited the formation of tube-like structures of HUVEC in Matrigel by 44%. In a cell culture model of angiogenesis consisting of a co-culture of endothelial cells with fibroblasts, transfection of antisense ONs resulted in an inhibition of tube formation of 61%. In conclusion, alpha v antisense ONs are potent inhibitors of angiogenesis in vitro. They might, therefore, be a therapeutic alternative to antagonists, which directly bind to alpha v integrins, and might be useful for the treatment of malignant tumors and hematological malignancies. PMID- 12082460 TI - Immunotherapy for murine K1735 melanoma: combinatorial use of recombinant adenovirus expressing CD40L and other immunomodulators. AB - We have constructed and tested five recombinant adenoviruses (Ads) that express a variety of immunomodulators, including CD40 ligand (CD40L), a potent costimulator of several components of the immune system. We demonstrate that CD40L expressed from Ad in K1735 mouse melanoma cells leads to a strong reduction in tumorigenicity and to efficient protective immunity in a vaccination setting. Subsequently, using a therapeutic approach, we found that local, intratumoral coinjection of CD40L- and IL-2-expressing Ads was superior to any other agents tested and resulted in an at least 1.9-fold increase in mean survival time, in contrast to systemic application of recombinant CD40L or GM-CSF proteins, which had no significant effects. When using vaccination as a therapeutic approach, the combinations of CD40L plus IL-2 or GM-CSF plus IL-2 from Ad gave rise to an extended (2.8-fold) increase in mean survival time. A detailed analysis of immune cells present within regressing tumors indicated that mainly CD4(+) and CD8(+) T cells, and to a lesser extent dendritic cells, infiltrated the tumor mass, but not NK cells, macrophages, or granulocytes. These results propose that a combination of CD40L plus IL-2 has an improved efficacy over the use of single agents when applied for direct in situ therapy or vaccination therapy. PMID- 12082461 TI - Tumor cell-targeting by phage-displayed peptides. AB - We isolated cancer cell-specific phages by subtracting and selecting complex peptide display phage libraries on cultured human cancer cells. The best candidate was selected by performing three rounds of subtraction before each of five selections on the human colorectal WiDr cell line. The phage showed more than 1000-fold higher binding efficiency for WiDr cells when compared to five other human cancer cell lines, including two of colorectal origin, and when compared to wild-type M13 phage. Fifty-fold higher binding efficiency was also seen for a human breast cancer cell line. We show that the WiDr cell binding of the selected phage was efficiently competed by the synthetic peptide HEWSYLAPYPWF, predicted from the phage sequence. This confirms that the specificity of the peptide is independent of the display by the phage coat proteins. The identified peptide may target biomarkers linked to colorectal cancer, and thus be useful for designing gene transfer vectors as well as diagnostic and prognostic tools for this disease. PMID- 12082462 TI - Protocol for gene transduction and expansion of human T lymphocytes for clinical immunogene therapy of cancer. AB - In preparation of a clinical phase I/II study in renal cell carcinoma (RCC) patients, we developed a clinically applicable protocol that meets good clinical practice (GCP) criteria regarding the gene transduction and expansion of primary human T lymphocytes. We previously designed a transgene that encodes a single chain (sc) FvG250 antibody chimeric receptor (ch-Rec), specific for a RCC tumor associated antigen (TAA), and that genetically programs human T lymphocytes with RCC immune specificity. Here we describe the conditions for activation, gene transduction, and proliferation for primary human T lymphocytes to yield: (a) optimal functional expression of the transgene; (b) ch-Rec-mediated cytokine production, and (c) cytolysis of G250-TAA(POS) RCC by the T-lymphocyte transductants. Moreover, these parameters were tested at clinical scale, i.e., yielding up to 5-10 x 10(9) T-cell transductants, defined as the treatment dose according to our clinical protocol. The following parameters were, for the first time, tested in an interactive way: (1) media compositions for production of virus by the stable PG13 packaging cell; (2) T-lymphocyte activation conditions and reagents (anti-CD3 mAb; anti-CD3+anti-CD28 mAbs; and PHA); (3) kinetics of T lymphocyte activation prior to gene transduction; (4) (i) T-lymphocyte density, and (ii) volume of virus-containing supernatant per surface unit during gene transduction; and (5) medium composition for T-lymphocyte maintenance (i) in between gene transduction cycles, and (ii) during in vitro T-lymphocyte expansion. Critical to gene transduction of human T lymphocytes at clinical scale appeared to be the use of the fibronectin fragment CH-296 (Retronectin) as well as Lifecell) X-fold cell culture bags. In order to comply with GCP requirements, we used: (a) bovine serum-free human T-lymphocyte transduction system, i.e., media supplemented with autologous patients' plasma, and (b) a closed cell culture system for all lymphocyte processing. This clinical protocol routinely yields 30-65% scFvG250 ch-Rec(POS) T lymphocytes in both healthy donors and RCC patients. PMID- 12082463 TI - 2-5A antisense therapy directed against human telomerase RNA inhibits telomerase activity and induces apoptosis without telomere impairment in cervical cancer cells. AB - Human telomerase RNA (hTR), an important component of telomerase, is a possible target of telomerase-based cancer gene therapy. The present study was undertaken to assess the efficacy of antisense hTR therapy using newly developed 2-5A (5' phosphorylated 2'-5'-linked oligoadenylate)-linked oligonucleotides against cervical cancer cells. ME180 and SiHa cells were treated with 2-5A-linked antisense hTR designed to complement the region of hTR between residues 76 and 94. The hTR expression, telomerase activity, cell viability, and apoptosis were then examined. The 2-5A anti-hTR effectively degraded hTR and inhibited telomerase activity. The 2-5A mutant anti-hTR and the anti-hTR without 2-5A were not capable of inhibiting telomerase activity. Inhibition of telomerase by 2-5A anti-hTR rapidly decreased cell viability only in telomerase-positive cells within 3-6 days after the treatment, when telomere length has not yet been shortened. This inhibition was associated with apoptosis, possibly through activation of caspase family members. These findings suggest that 2-5A-linked antisense-hTR therapy has a potent telomerase-inhibitory effect associated with a cytocidal effect from caspase-induced apoptosis, and may therefore be a potential tool in telomerase-based gene therapy against cervical cancers. PMID- 12082465 TI - Sorting out the ways to sort out fetal growth. PMID- 12082466 TI - Evaluation of the direct antiglobulin (Coombs') test for identifying newborns at risk for hemolysis as determined by end-tidal carbon monoxide concentration (ETCOc); and comparison of the Coombs' test with ETCOc for detecting significant jaundice. AB - OBJECTIVE: First, to determine the sensitivity, specificity, and positive predictive value (PPV) of the direct antiglobulin test (DAT) for significant hemolysis in the neonate, as referenced to end-tidal carbon monoxide, the criterion standard for estimating the rate of hemolysis; and second, to evaluate the predictive value of the two procedures for significant jaundice. DESIGN: Consecutive term newborns admitted to the nursery of an inner-city university hospital over a 15-week period. DAT screening by the Blood Bank was performed on all. End-tidal carbon monoxide levels were obtained at 12+/-6 and at 24+/-6 hours of age. Infants of nonsmoking mothers whose 12-hour exhaled carbon monoxide level was > or = 95th percentile were defined as having significant hemolysis. RESULTS: n=660; DAT was positive in 23 (3.5%). Using the 12-hour end-tidal carbon monoxide > or = 3.2 microl/l (> or = 95th percentile) as reference (n=499 nonsmokers), the sensitivity of the DAT was 38.5% (10 of 26) and specificity 98.5% (466 of 473) for the detection of significant hemolysis. The PPV of the DAT for significant hemolysis at 12 hours was 58.8% (10 of 17). For significant jaundice the PPV of end-tidal carbon monoxide was greater than that for DAT (65.4% vs 52.9%), although not statistically so (p=0.25). The negative predictive values were similar. CONCLUSION: DAT fails to identify over half of the cases of significant hemolysis that are diagnosed by end-tidal carbon monoxide. A neonate with a positive DAT has about a 59% chance of having significant hemolysis. End-tidal carbon monoxide may also provide a more sensitive index for predicting significant jaundice. PMID- 12082467 TI - Mean arterial pressures during the first 24 hours of life in < or = 600-gram birth weight infants. AB - OBJECTIVE: To determine mean arterial pressure values during the first 24 hours for "stable" and "unstable" extremely low birth weight (ELBW) infants and to ascertain its association with perinatal factors. BACKGROUND: In ELBW infants, hypotension is diagnosed by nonspecific clinical signs together with reference arterial pressure values extrapolated from regression models or from scarce actual observations. DESIGN: Retrospective cohort study. METHODS: 101 ELBW (< or = 600 g) infants born in our medical center (1989-2000). Considered stable were 36 infants with umbilical cord hemoglobin > or = 14 g/dl who, although mechanically ventilated, had normal acid-base balance, no patent ductus arteriosus, had not received indomethacin, steroids, muscle relaxants, narcotics, were never treated for hypotension and survived at least 7 days. The remaining 65 infants constituted the unstable group. Arterial pressures were determined by oscillometry (OBP) and direct transducer readings through an umbilical line (MAP). All admission and 10% of the readings were by OBP; the remaining 1877 measurements were by MAP. RESULTS: Stable and unstable infants were similar in birth weight, demographics, history of chorioamnionitis, antepartum steroids, low Apgar scores, administration of epinephrine during resuscitation, and sepsis. Stable infants were different from unstable in gestational age (27+/-2 vs 25+/-2 weeks' gestational age [w GA]), history of preterm labor, preeclampsia, and neonatal mortality (22 vs 68%). Admission OBP (30+/-7 vs 29+/-10 Torr) were similar and 1-hour MAP were different (30+/-6 vs 27+/-7 Torr) between both groups. MAP for stable infants was higher throughout the 24 hours. Greater differences were noted between 3 and 6 hours when 34 of 65 unstable infants were treated for hypotension. Mean MAP and 10 percentile values for stable infants at 1, 3, 6, 12, and 24 hours were 30 (22), 31 (24), 32 (25), 34 (24), and 35 (28) Torr, respectively. MAPs did not correlate with birth weight, but they were lower among 19 stable infants < or = 26 w GA than among 17 stable infants > or = 27 w GA. History of preeclampsia, antenatal steroids, intratracheal epinephrine and cord hemoglobin did not influence MAP. Low 1-minute Apgar score and intracranial hemorrhage were associated with low MAP during the first day. CONCLUSION: There is a wide variation of GA among ELBW infants. MAPs increase with GA and with postnatal age. Shortly after birth, arterial pressures are similar for stable and unstable infants. Failure to increase MAP between 3 and 6 hours of life should create concern. MAP < or = 28 Torr at 3 hours of life is a reasonable, but not absolute, predictor of the need for hypotension treatment. PMID- 12082468 TI - Human milk reduces outpatient upper respiratory symptoms in premature infants during their first year of life. AB - OBJECTIVE: To determine if ingestion of human milk after discharge reduces symptoms of infections in premature infants. STUDY DESIGN: Follow-up of 39 infants with birth weights <2000 g, 24 of whom received any amount of human milk and 15 of whom received only formula after discharge, was carried out. Mothers were given a calendar on which they recorded any signs of infections and feeding and day-care information. Data were collected at 1 month after discharge and at 3, 7, and 12 months corrected age. RESULTS: Results show no differences between groups in birth weight, gestation, gender, maternal age, parental tobacco use, number of siblings, and day-care attendance. Socioeconomic status score was higher in the human milk group. Infants who received human milk had fewer days of upper respiratory symptoms at 1 month after discharge (p<0.025) and at 7 months corrected age (p<0.025). CONCLUSION: Ingestion of human milk post discharge is associated with a reduction of upper respiratory symptoms in premature infants during their first year of life. PMID- 12082469 TI - Reliability of conventional and new pulse oximetry in neonatal patients. AB - OBJECTIVES: Pulse oximetry is widely used in the NICU, but clinicians often distrust the displayed values during patient motion, i.e., questionable oxygen saturation (SpO(2)) and pulse rate (PR) values. Masimo Corporation (Irvine, CA) has developed pulse oximetry with claims of resistance to sources of interference. To test this premise, we compared the performance of the Masimo SET pulse oximeter to a conventional device, Nellcor N-200, and then with three other new-generation pulse oximeters, Nellcor N-395, Novametrix MARS, and Philips Viridia 24C. STUDY DESIGN: We studied 26 nonsedated NICU infants who were on supplemental oxygen and/or mechanical ventilation. ECG heart rate (HR) from a bedside monitor and SpO(2) and PR from the two pulse oximeters were captured by a PC for a total of 156 hours. The ECG HR and pulse oximeter spectral waveform were analyzed at alarms for hypoxemia (SpO(2)< or = 85%) and/or bradycardia (HR< or = 80 bpm). We then compared the performance of the Masimo SET to three other new generation pulse oximeters, Agilent Viridia 24C, Nellcor N-395, and Novametrix MARS, in a similar population of seven infants for a total of 28 hours. We added to the test criteria the ability of the various pulse oximeters to track acute changes in HR. RESULTS: Compared with Nellcor, Masimo SET had 86% fewer false alarms, which also were shorter in duration, resulting in 92% less total alarm time. Masimo SET also identified nearly all bradycardias versus 14% for the Nellcor. Compared with the new-generation pulse oximeters, false desaturations, data drop-outs, and false bradycardias were lowest for Masimo SET, as was the capture of true desaturations and bradycardias. Notably, the new-generation devices differed greatly in their ability to detect changes in HR (i.e., the frequency of frozen PR during times of ECG HR change was 0, 6, 11, and 46 for Masimo, Nellcor, Philips, and Novametrix, respectively). CONCLUSIONS: Masimo SET pulse oximetry recorded markedly fewer false SpO(2) and PR alarms and identified more true hypoxic and bradycardic events than either conventional or other new generation pulse oximeters. Masimo SET also most closely reflected the ECG rate irrespective of accelerations or decelerations in HR. SPECULATION: Routine use of Masimo SET pulse oximetry in the NICU could improve clinician confidence in the parameter leading to more judicious titration of oxygen with possible reductions in hypoxic (e.g., pulmonary hypertension) and hyperoxic (e.g., retinopathy of prematurity) pathology. Additionally, a more trustworthy technology should equate with fewer confirmatory arterial blood gas analyses (less blood loss), and faster weaning from the mechanical ventilation (less chronic lung disease). PMID- 12082470 TI - Continuous blood gas monitoring using an in-dwelling optode method: clinical evaluation of the Neotrend sensor using a luer stub adaptor to access the umbilical artery catheter. AB - INTRODUCTION: Arterial blood gases are essential in the management of critically ill neonates. A new technology using the Neotrend system (Diametrics Medical) allows for continuous measurement of pH, PaCO(2), and PaO(2), and calculates oxygen saturation, bicarbonate, and base excess. OBJECTIVE: To evaluate and compare the results of continuous blood gas measurement using the Neotrend system with a standard system of blood gas analysis in our intensive care unit. DESIGN: Prospective, controlled, interventional study. SETTING: The neonatal intensive care unit of a tertiary referral center. PATIENTS: Neonates with respiratory distress who required respiratory support and frequent arterial blood gas sampling and had a UAC. RESEARCH DESIGN AND METHOD: To enable Neotrend sensor access to an existing Argyle umbilical artery catheter (UAC) the catheter was cut at the 25-cm mark and connected to an 18-gauge blunt needle luer stub adaptor (Vygon 95440). The study began with the insertion of the Neotrend sensor. Subjects remained on the study until the UAC was discontinued and/or frequent blood gases were no longer needed. The blood gas results from the Neotrend system were not used in clinical management of the patient. BLOOD GAS MEASUREMENT: During the study period, with each blood sample drawn for laboratory analysis, a printout from the Neotrend monitor was recorded for comparison. RESULTS: A total of 217 pairs of blood gas samples were collected from seven neonates. The mean bias/precision for pH was 0.01/0.04; for PaO(2) 0.72/18.5 mm Hg; and for PaCO(2) 3.96/2.63 mm Hg. The correlation (r value) between the sensor reading and the blood gases were 0.85 for pH, 0.96 for PaO(2), and 0.92 for PaCO(2). CONCLUSION: The blood gases compared in the two methods had a strong correlation for pH, PaCO(2), PaO(2), and oxygen saturation. Although the bicarbonate and base excess values showed suboptimal statistical correlation, the difference was not clinically relevant. Results of this study indicate that this technology provides an accurate means of monitoring continuous blood gas parameters in neonatal patients. It also allows reduced healthcare provider exposure to blood and decreased patient iatrogenic blood loss. PMID- 12082471 TI - Protease inhibitors and decreased birth weight in HIV-infected pregnant women with impaired glucose tolerance. AB - OBJECTIVE: To assess the impact of highly active antiretroviral therapy, including a protease inhibitor (HAART/PI), on maternal glucose tolerance and fetal growth. STUDY DESIGN: A retrospective chart review of pregnancy outcomes of human immunodeficiency virus-infected pregnant women receiving HAART/PI (n=41) or zidovudine monotherapy (n=23). RESULTS: Abnormal 1-hour glucose tolerance tests (1hGTT) were observed in 30% of subjects receiving HAART/PI between 24 and 28 weeks' gestation. An elevated 1hGTT was associated with a significantly lower mean birth weight in subjects receiving HAART/PI compared to babies born to mothers with a normal 1hGTT (3.40+/-0.09 vs 3.00+/-0.18 kg, p<0.05, ANOVA). CONCLUSION: HAART/PI therapy is associated with an increased rate of impaired glucose tolerance in pregnancy and impaired fetal growth. This finding merits further investigation. PMID- 12082472 TI - Comparison of kangaroo care and standard care: behavioral organization, development, and temperament in healthy, low-birth-weight infants through 1 year. AB - OBJECTIVE: To determine whether Kangaroo Care (KC) for healthy, low-birth-weight (LBW) infants can promote better behavioral and developmental outcomes. STUDY DESIGN: In this historical control study, 26 infants in the KC group (GA: 34.3+/ 2.5 weeks, BW: 1833.9+/-167.6 g) and 27 infants in the comparison group who received the standard medical-nursing care (34.6+/-2.3 weeks, 1850.9+/-156.7 g) were analyzed by the Neonatal Behavioral Assessment Scale (NBAS) at 40 weeks of postmenstrual age, the Bayley Scales of Infant Development and the Carey's Infant Temperament Questionnaire (ITQ) at 6 and 12 months corrected ages. RESULTS: KC infants had significantly higher NBAS scores in Orientation, State Regulation, and Supplementary items; lower Intensity scores and higher Mood scores at 6 months on the ITQ; and higher Bayley Scales score at 12 months. CONCLUSION: KC effectively promoted neonatal behavioral organization and enhanced developmental outcome over the first year of life for LBW infants. PMID- 12082473 TI - Impact of family health insurance and other environmental factors on universal hearing screen program effectiveness. AB - OBJECTIVE: The study objective was to evaluate the relationship among health insurance type, other demographic factors, and newborn hearing screen compliance and outcomes. STUDY DESIGN: The cohort consisted of 39,153 infants screened in Rhode Island between July 1, 1995 and June 30, 1998. Multivariate analyses were completed to evaluate relationships between health insurance type and completion of the in-hospital hearing screen and the rescreen, if indicated. RESULTS: Successful newborn screen rates ranged from 98.1% to 99.8%. Infants with traditional Medicaid insurance were more likely to not be screened (p<0.0001) and to not return for a rescreen (p<0.0001). Infants in families with managed care Medicaid had screen compliance similar to infants with commercial health insurance. Multivariate analyses revealed that Medicaid insurance, no insurance, neonatal intensive care unit status, and out-of-state address predicted no initial screen (p<0.001) and no rescreen (p<0.0001). CONCLUSION: In population based health services, it is important that the effects of socioeconomic and demographic variables on outcomes be evaluated. PMID- 12082474 TI - Positive changes among very low birth weight infant Apgar scores that are associated with the Neonatal Resuscitation Program in Illinois. AB - OBJECTIVE: The American Academy of Pediatrics and American Heart Association sponsored Neonatal Resuscitation Program (NRP) was started in Illinois 1987 to standardize knowledge and skills for successful resuscitation to decrease newborn morbidity and mortality. This study evaluated the hypothesis that the statewide NRP in Illinois hospitals was associated with positive outcomes in the Apgar scores among very low birth weight newborns. STUDY DESIGN: A retrospective design was used to compare the (a) different rates of lower (0-6) and higher (7-10) 1- and 5-minute Apgar scores and (b) positive change from a lower 1-minute to a higher 5-minute Apgar score before and after NRP was started. Analyses included maternal characteristics, birth weight groups (grams: 500-749, 750-999, 1000 1249, 1250-1499), and hospital levels (I, II, III). RESULTS: A significantly higher proportion of infants had a higher 1-minute and a higher 5-minute Apgar score after NRP, overall, for Level II and II+ hospitals and for each weight group. Logistic regression indicated that newborns with low 1-minute Apgar scores were 81% more likely to have a higher 5-minute Apgar score after NRP. CONCLUSION: Significant improvement in Apgar score occurred after NRP. Empirical support is demonstrated for the clinical effectiveness of NRP instruction in Illinois hospitals. PMID- 12082475 TI - Comparison of the perinatal morbidity and mortality of the presenting twin and its co-twin. AB - OBJECTIVE: To compare the perinatal outcome of the presenting twin to its co twin. STUDY DESIGN: All live nonanomalous twin gestations delivered at >25 weeks' gestation between 1984 and 1994 (N=461) were identified. Twin A was compared to twin B regarding the following variables: presentation, Apgar score (AS) <4 at 1 minute, AS <7 at 5 minutes, birth weight, gender, traumatic delivery, meconium stained amniotic fluid, cord prolapse, need for mechanical ventilation, intraventricular hemorrhage, respiratory distress syndrome, sepsis, seizures, perinatal mortality, and length of nursery stay. RESULTS: Except for differences in presentation, the perinatal outcome was similar in both twins regarding variables studied. This continued to hold true after subdividing according to mode of delivery, when infants with birth weight < or = 1500 g were considered separately, and when vaginally delivered cephalic twin A was compared to the noncephalic co-twin. However, differences in some outcome variables became evident when pregnancies with only one affected member were analyzed separately. CONCLUSIONS: When all twin pairs are considered, the outcome of the second-born twin is similar to that of the first-born regardless of the mode of delivery, presentation, or birth weight < or = 1500 g. If only those twin pairs with one affected and one unaffected twin are considered, it becomes apparent that the presenting twin is at increased risk for infection-related morbidities whereas the co-twin is at risk for other complications. PMID- 12082476 TI - Neonatal anthropometric measurements to predict birth weight by ultrasound. AB - OBJECTIVE: To develop a more accurate ultrasound birth weight (BW) model using neonatal anthropometric measurements. STUDY DESIGN: Two hundred thirty-one newborns were evaluated. Measurements included weight; head, chest, and abdominal circumferences (umbilicus and liver), humerus, and femur lengths. Infants were randomly assigned into two groups (G(1) and G(2)). Anthropometric measurements that are obtainable by ultrasound were generated from G(1). Stepwise regression and a bootstrap analysis were used to create the prediction models. The models were validated using G(2). RESULTS: The final stepwise regression model included FL and circumferences of the head, chest, and abdomen. The correlations were: G(1): R(2)=0.91, p<0.001; G(2): R(2)=0.90 p<0.001. There was no difference between derived and actual BW in G(1) (p=0.42) or G(2) (p=0.28). The mean absolute percent error between the prediction model and actual BW was 3.8%. CONCLUSION: Neonatal anthropometric models are strongly predictive of actual BW. This model will be tested prospectively using ultrasound to predict fetal weight. PMID- 12082477 TI - Glyburide compared to insulin for the treatment of gestational diabetes mellitus: a cost analysis. AB - OBJECTIVE: To compare the costs associated with glyburide compared to insulin for the treatment of gestational diabetes unresponsive to dietary therapy. STUDY DESIGN: A cost model was designed. The model excluded costs that were identical for both treatment arms, such as the cost of monitoring glucose control. Insulin treatment costs included average wholesale drug costs, wholesale delivery costs (syringes, alcohol pads), and costs of office staff educating patients. Glyburide costs were based on average wholesale drug costs. Downstream costs of potential inpatient evaluation for hypoglycemia were included in the model. RESULTS: In our baseline model, glyburide was significantly less costly than insulin for the treatment of gestational diabetes. The average cost saving per patient based on wholesale drug costs and hospital costs was US$165.84. Actual retail drug savings and hospital charge savings are potentially considerably greater. The strongest determinant of cost savings was medication cost. The model was less sensitive to the one-time costs of inpatient treatment and patient education. CONCLUSION: Glyburide is less costly than insulin for the treatment of gestational diabetes. Cost models can be useful to physicians deciding between two equally efficacious medications, allowing them to incorporate information about their individual practice styles with a complex balance of cost implications. PMID- 12082479 TI - Using mnemonics and visual imagery to teach the new neonatal resuscitation program. AB - Mnemonics have long been used to help learners remember facts in various disciplines of medicine. With the introduction of the revised Neonatal Resuscitation Program (NRP), it is vital for learners to know and remember important changes to the NRP curricula. While teaching the new curricula, we found that learners appeared confused, being unable to provide the correct sequence of answers to the questions posed during the assessment phase of the course. Mnemonics were developed to aid memory recall and optimize resuscitation skills. PMID- 12082478 TI - Chorioamniotic membrane separation following fetal surgery. AB - OBJECTIVE: As the volume of fetal surgery cases has steadily increased, an increasing incidence of chorioamniotic membrane separation (CMS) has been noted. Due to the potential adverse consequences from this abnormality, we reviewed the last decade of experience with fetal intervention at our institution and examined the incidence and outcomes of fetuses given this diagnosis. STUDY DESIGN: A retrospective chart review of 75 fetal surgery cases at our institution was performed. Variables analyzed included preoperative, operative, and outcome data. Postoperative ultrasounds were evaluated for the presence of CMS. RESULTS: Excluding operative deaths, the incidence of CMS was 47%. There were significant differences (p<0.05) in time to delivery (7 vs 5 weeks), cases using a perfusion pump (80% vs 60%), and number of trocars (2.13 vs 1.54) in cases of CMS versus those without. Ultrasounds showed normal to high levels of amniotic fluid in 97% of cases. There was an increased incidence of premature rupture of membranes (63% vs 45%), preterm labor (57% vs 38%), and chorioamnionitis (29% vs 15%) with CMS, but no difference in mortality rate. CONCLUSION: CMS is a frequent finding following fetal surgery. It is associated with significant morbidity but is manageable with close follow-up in a hospital setting. Following fetal surgery, the finding of CMS can be a life-threatening complication that warrants further study to understand its etiology and prevention. PMID- 12082480 TI - Intracranial lipomas in neonate. PMID- 12082481 TI - The use of topical nitroglycerin ointment to treat peripheral tissue ischemia secondary to arterial line complications in neonates. AB - Catheterization of the aorta through the umbilical artery and/or of peripheral arteries in neonates may be accompanied by a number of complications, of which thrombotic phenomena and peripheral vasospasm are the most common. Two neonates with peripheral ischemia caused by vasospasm from indwelling umbilical artery catheterization and one infant with left hand ischemia due to a left radial artery line were successfully treated with 2% nitroglycerin ointment. No adverse effects were noted. PMID- 12082482 TI - Tissue-limited mosaicism in Pallister-Killian syndrome -- a case in point. AB - We report a case of Pallister-Killian syndrome in a term female infant. Antenatal ultrasound showed left diaphragmatic hernia and polyhydramnios. She was ventilated from birth and the diaphragm defect repaired on day 5. She had dysmorphic features, including median cleft palate, patchy frontotemporal alopecia, hypopigmented skin whorls, and bilateral profound sensorineural hearing loss. Fetal and postnatal karyotypes of peripheral lymphocytes were both normal, 46, XX. Subsequently, a skin fibroblast culture showed mosaic tetrasomy of isochromosome 12p both on G-banding and fluorescence in situ hybridization, consistent with Pallister-Killian syndrome. This case illustrates the importance of using the appropriate sample type for karyotype analysis with implications for prenatal and postnatal diagnosis. PMID- 12082483 TI - Multiple arterial thrombi and in utero leg gangrene in an infant of a diabetic mother. AB - This case report is of an infant of a diabetic mother who presented with lower extremity gangrene occurring in utero that necessitated a below the knee amputation at 3 hours of age. The association of venous thrombi in infants of diabetic mothers (IDMs) has been firmly established. However, in this case the thrombi were arterial and resulted in gangrene during the second trimester of gestation. The reason for the increased tendency to develop thrombi in infant of diabetic mothers has not been elucidated. We did an extensive workup of this infant to look for a cause of the arterial thrombus. The only abnormality found was in the plasminogen activity, which was significantly lower than the reference values for infants of the same gestational and chronological age. The significance of this finding on the propensity of IDMs to develop thrombi is discussed. PMID- 12082484 TI - Group B streptococcal sepsis. PMID- 12082487 TI - Losartan and the meaning of LIFE. PMID- 12082488 TI - The effect of antihypertensive drugs on the fetus. AB - A critical review of the literature on the effects of antihypertensive drugs on the fetus in pregnant women is presented. The survey covers the alpha-adrenergic receptor agonists, beta-blockers including topical eye medications, alpha-beta blockers, calcium antagonists, diuretics, and angiotensin-converting enzyme (ACE) inhibitors. The lack of data on angiotensin II receptor blockers is noted although effects are considered to be similar to those reported with ACE inhibitors and therefore to be avoided. Analysis of the literature underscores that some antihypertensive drugs can be used safely at certain stages of pregnancy, while others are suspect and to be avoided at all costs. The lack of placebo-controlled studies on the treatment of severe hypertension in pregnancy due to ethical considerations is discussed against the background of the pressing need to treat these women despite the possible deleterious effects of antihypertensive drugs. PMID- 12082489 TI - Smoking trends in hypertensive and normotensive Finns during 1982-1997. AB - Despite the evidence that smoking is one of the main predictors of the cardiovascular disease risk among hypertensive subjects, there are very few data available of the smoking trends of these subjects at public health level. This study assesses the trends in smoking and in smoking cessation advice given by physicians in the hypertensive and normotensive population in Finland during 1982 1997. The data were derived from four independent cross-sectional standardised population surveys conducted in 1982, 1987, 1992 and 1997 in the provinces of North Karelia and Kuopio in eastern Finland and the region of Turku-Loimaa in south-western Finland. Men and women aged 25-64 years, selected randomly from the national population register, were classified to four groups according to their blood pressure level and antihypertensive treatment status: normotensive, unaware hypertensive, aware but not treated hypertensive and treated hypertensive. The total number of participants was 24 083. In men, the prevalence of smoking decreased significantly in both treated hypertensive patients and normotensive subjects during 1982-1997, whereas it increased significantly in treated hypertensive women. The proportion of current smokers who had been advised to stop smoking by their physician was significantly higher in treated hypertensive men compared to the other subgroups of men (P < 0.001). The observed decreasing trend in smoking in men is encouraging, but the increase in smoking among hypertensive women taking antihypertensive medication warrants concern. In the future, the methods used for smoking cessation advice given by health care personnel should be intensified to make this effort more effective. PMID- 12082490 TI - Distribution of blood pressure and prevalence of hypertension in Tehran adult population: Tehran Lipid and Glucose Study (TLGS), 1999-2000. AB - The purpose of this study was to estimate the current prevalence and distribution of hypertension in an adult Tehranian population. Data were collected for 3343 men and 5148 women aged 20-69 years in the Tehran Lipid and Glucose Study (TLGS), which is a cross-sectional phase of a large epidemiologic study, first established in 1999. The study used the mean of two separate blood pressure (BP) measurements in each individual. Twenty-two percent (23% of women vs 20% of men, P = 0.01) had hypertension according to 'JNC-VI' and 'WHO-ISH' criteria. The average systolic BP (SBP), diastolic BP (DBP) and pulse pressure of hypertensive participants were 31, 16, and 15 mm Hg higher than the corresponding value for normotensives, respectively. Thirty-six percent of participants with JNCVI-based hypertension were using antihypertensive medication (23% of men and 43% of women). Of these, 40% (45% of men and 39% of women) had normal BP. Hypertension awareness was 50% in these participants (57% in men vs 37% in women, P < 0.001). Data for 3179 men and 4646 women aged 20-69 years with no antihypertensive treatment were used for analysis of BP measures. Of these, 15% (16% of men and 14% of women, P = 0.006) had high and 85% (84% of men and 86% of women) normal or high-normal BP levels according to JNC-VI. Prevalence of optimal BP was 49% (47% of men and 51% of women). Mean SBP was 117.8 +/- 16.6 and 116.4 +/- 16.4 mm Hg in men and women, respectively (P < 0.001). The equivalent values were 77.4 +/- 10.7 and 77.3 +/- 9.9 mm Hg for DBP (P = 0.5) and 40.4 +/- 12 and 39.1 +/- 11.7 mm Hg for pulse pressure (P < 0.001). A relatively high prevalence of JNC-VI/WHO-ISH defined hypertension was found in the TLGS adult population with 50% undiagnosed and 60% uncontrolled hypertension. These findings emphasise further considerations for detection and better management of hypertension in the urban population of Tehran. PMID- 12082491 TI - Hypertension control and medication increase in primary care. AB - Over half of treated patients with hypertension are not well controlled. However, little is known about physicians' prescribing behaviour for these patients. Our objective was to clarify whether physicians increase antihypertensive medication in patients with poorly controlled hypertension and what characteristics are predictors of medication increase. This was a retrospective cohort study by surveying medical records in primary care clinics in Tochigi, Japan. Twenty-nine of 79 randomly selected physicians agreed to select 20 consecutive hypertensive patients. This resulted in 547 patients (women 60%; mean (s.d.) age, 68 (12) years) who had blood pressure measurements taken in 1998 and prescription of antihypertensive medication in 1998 and 1999. Mean (s.d.) systolic/diastolic blood pressure was 142 (12)/81 (9) mm Hg and the percentage of patients in good control (<140/90 mm Hg), fair (140-159/90-94) and poor (> or =160/95) were 42%, 47%, and 11%, respectively. Physicians increased medication in 28% of poorly controlled patients (95% confidence interval (CI), 17-41%), which was more than those in fair (12%, 95%CI 8-16%) or good control (7%, 95%CI 4-12%). Multivariate logistic regression analysis showed that systolic and diastolic blood pressures were positively, and the number of kinds of antihypertensive medications and the age of the physician were negatively, associated with an increase in medication. In conclusion, primary care physicians did not increase antihypertensive medication adequately for patients with uncontrolled hypertension. Attempts to understand and to change physicians' prescription behaviour could reduce the burden of uncontrolled hypertension among treated hypertensive patients. PMID- 12082492 TI - Cardiovascular and renal responses to mental challenge in highly and moderately active males with a family history of hypertension. AB - The objective of this study was to compare FBF and renal responses to mental challenge in highly and moderately active males with a family history of hypertension. Normotensive, healthy males with a family history of hypertension (n = 18) were recruited into moderately active and highly active groups. Cardiovascular, FBF, and renal responses to a 10-min Stroop mental challenge were measured. Urine was analysed for levels of sodium and potassium pre and post stressor as an indicant of renal blood flow. The results were that the moderately active males demonstrated a significantly higher level of FBF reactivity to mental challenge compared with that of the highly active. Heart rate change and FBF change during the stressor were positively correlated (r = 0.75, P < 0.01). Both groups, however, demonstrated a similar pattern of sodium excretion to mental challenge. These findings suggest that physical activity level is associated with FBF reactivity but not renal reactivity to mental challenge in offspring hypertensives. That sodium excretion was no different post-stressor in the moderately active group suggests that the exaggerated forearm vasodilatation response was not due to renal vasoconstriction. PMID- 12082493 TI - Assessing resting heart rate in adolescents: determinants and correlates. AB - The aim of this study was to evaluate the distribution of resting heart rate and its biological and environmental determinants in adolescents. The study was cross sectional and the population consisted of 2230 children and adolescents, age range 12-18 years, enrolled randomly from state schools in Turin, Italy. In all participants the following parameters were evaluated: heart rate, blood pressure (BP), weight, height, degree of sexual development, physical activity, parental socio-cultural level. Heart rate and BP were measured after 5, 10 and 15 min in a sitting position. Furthermore, to obtain regression equations to define heart rate as a function of the other variables available, a multiple regression analysis was performed. In both sexes BP, but not heart rate, declined significantly from the first to the last determination. Heart rate was positively and significantly correlated to BP level in both sexes; heart rate was higher in girls (3 bpm) and followed a progressive decreasing trend with age in both sexes, that was opposite to BP values. Age, sexual maturation, height, physical activity and parental socio-cultural level were independent determinants of resting heart rate. In conclusion, resting heart rate in adolescents is related to several methodological, constitutional and environmental factors that have to be taken into account when assessing heart rate values and constructing tables of normal values. PMID- 12082494 TI - Oxidative stress in malignant and non-malignant phase hypertension. AB - Malignant hypertension (MHT) is a rare and severe form of hypertension characterised by arteriolar necrosis and severe vascular damage, leading to stroke, myocardial infarction and death. We hypothesised that in addition to endothelial damage, MHT may be associated with increased oxidative stress. Lipid hydroperoxides (LHP, an index of oxidative damage) and plasma von Willebrand factor (vWf, an index of endothelial damage/dysfunction) were measured in 16 patients with MHT and compared with 16 non-malignant essential hypertensives and 32 normotensive controls. vWf was greater in MHT (mean 117 iU/dL) than in non malignant hypertensives (97 iU/dL) or normotensive controls (100 iU/dL) (ANOVA P = 0.017). However, although LHP were greater in MHT (mean 10.6 micromol/L) than in normotensives (4.5 micromol/L, P < 0.001), the levels in MHT were similar to those in non-malignant hypertension (12.3 micromol/L). In conclusion endothelial damage (raised vWf) was more evident in MHT compared with both normotensive controls and with non-malignant hypertension, whilst oxidative stress (raised LHP) was increased to a similar extent in both hypertension groups when compared with normotensive controls. These observations raise the possibility abnormal oxidative stress is probably not the mechanism responsible for the endothelial damage seen in malignant phase hypertension. PMID- 12082495 TI - Serum cholesterol affects blood pressure regulation. AB - A close relationship between abnormalities of the lipid metabolism and arterial hypertension has been observed in several epidemiological studies. The aim of the present study was to investigate whether serum cholesterol might affect blood pressure (BP) levels at rest, during ambulatory monitoring or during sympathetic stimulation-independently of other variables such as body weight or serum insulin thus influencing the outcome of hypertensive complications. Seventy-three patients with sustained newly-discovered and never-treated hypertension were divided into tertiles according to their serum cholesterol levels and their resting BP, 24-h BP and BP during isometric exercise (handgrip) were compared. Cardiac mass and carotid wall thickness were measured by echographic technique. The results were that tertiles were similar for body weight, blood glucose and serum insulin, but different for serum cholesterol and triglycerides. BP at rest and during 24-h monitoring was similar in the three groups, whilst a significant difference was detected during sympathetic stimulation by handgrip, with systolic and diastolic BP increasing by 16/12, 28/19 and 30/23 mm Hg (P < 0.01) in lower, medium and higher tertiles, respectively. Intima-media layer of the carotid arteries was also significantly thickened in the groups with higher cholesterol levels (0.54 +/- 0.07, 0.67 +/- 0.14, 0.68 +/- 0.15, P < 0.05). These data support the conclusion that even in patients with recently discovered hypertension, cholesterol levels may influence the BP response to adrenergic stimulation as well as the outcome of target organ disease. PMID- 12082496 TI - Effects of alcohol intake on ambulatory blood pressure, heart rate, and heart rate variability in Japanese men with different ALDH2 genotypes. AB - The effects of alcohol intake on haemodynamics and heart rate variability were investigated with relation to genotypes of aldehyde dehydrogenase 2 (ALDH2), which were determined in 33 male Japanese volunteers (mean +/- s.e., 35.7 +/- 1.4 years) using the PCR-RFLP method. On the alcohol intake day, they consumed 660 ml of beer containing 33 ml of ethanol (0.3-0.5 g/kg of body weight) from 18.00 to 18.30. On the control day, they ingested the same amount of non-alcoholic beer. Ambulatory blood pressure, heart rate, and ECG R-R intervals were measured during a 24-h period with a portable recorder. A power spectral analysis of R-R intervals was performed to obtain the low-frequency (LF) and high-frequency (HF) components. Sixteen subjects were homozygotes for the normal ALDH gene (active ALDH2), only one was a homozygote for the mutant ALDH2 gene (inactive ALDH2), and the remaining 16 were heterozygotes (inactive ALDH2). Alcohol intake did not change 24-h average blood pressure (BP) either in the active ALDH2 group or in the inactive ALDH2 group. However, during the time interval from 18.30 to 0.00, alcohol intake significantly decreased diastolic BP in the active ALDH2 group and both systolic and diastolic BPs in the inactive ALDH2 group. In the active ALDH2 group, alcohol intake did not change heart rate, while in the inactive ALDH2 group, alcohol intake significantly increased 24-h average heart rate by 5.3 +/- 1.6 beats per minute (P < 0.01). In the active ALDH2 group, neither the LF nor the HF component was changed by alcohol intake, while in the inactive ALDH2 group, both the LF and the HF components were significantly decreased during the time interval from 18.30 to 0.00. These results demonstrate for the first time that ALDH2 genotypes modify the effects of intake of a small amount of alcohol on haemodynamics and heart rate variability in Japanese men. PMID- 12082497 TI - Increased red cell sodium-lithium countertransport and lymphocyte cytosolic calcium are separate phenotypes in patients with essential hypertension. AB - Increased red blood cell sodium-lithium countertransport (SLC) activity and elevated intracellular calcium have been observed in hypertensive patients. The association of these ion transport abnormalities with each other and with another phenotype, insulin resistance, has been suggested. We investigated whether elevated SLC activity and increased lymphocyte cytosolic calcium (Ca(cyt)) occur in the same individuals and whether either is associated with hyperinsulinaemia. We measured SLC activity, lymphocyte Ca(cyt)and fasting insulin levels in hypertensive patients and normal subjects. Consistent with prior studies, SLC activity was significantly and positively correlated with fasting insulin levels (r = 0.45, P < 0.01). However, SLC activity and lymphocyte Ca(cyt) were significantly but inversely correlated (r = -0.42, P < 0.01) and lymphocyte Ca(cyt) was also inversely correlated with fasting insulin (r = -0.55, P < 0.001). When the study participants were instead separated into two groups based on fasting insulin levels, those above the median (15 microU/ml) had significantly higher SLC activity and significantly lower Ca(cyt). When separated by lymphocyte Ca(cyt) levels (above or below 120 nM) those patients with low lymphocyte Ca(cyt) had significantly higher SLC activity and significantly higher insulin levels. Multiple linear regression showed that fasting insulin was significantly predictive of SLC activity (P = 0.05) and Ca(cyt) (P < 0.01). Thus, elevated SLC activity and increased lymphocyte Ca(cyt) are separate and distinct ion transport phenotypes in hypertensive patients, linked through a relationship to hyperinsulinaemia that is direct with SLC activity and inverse with lymphocyte Ca(cyt). PMID- 12082498 TI - Practice audits: reliability of sphygmomanometers and blood pressure recording bias. AB - It is well established that numerous errors, biases and omissions in recording blood pressure exist. This study had two objectives. Firstly, to measure the accuracy of sphygmomanometers used in general practice and secondly to assess digit bias in blood pressure recording. This study was carried out in the then Northfield PCG, which comprised 18 practices and 67 GPs. A total of 131 mercury and aneroid sphygmomanometers were tested for accuracy by a trained technician in accordance with the methods specified in BS 2743 (1990). Accuracy was defined as an error of greater than 10 mm Hg. The second part of the methodology involved undertaking an audit of the proportion of registered patients aged 35-80 years who had their blood pressure measured within the last 5 years by members of the Primary Health Care Team. The results were that of the mercury and aneroid sphygmomanometers tested, 17% were inaccurate. Of these, 4% recorded an error greater than 10 mm Hg. One percent of mercury and 10% of aneroid sphygmomanometers recorded an error greater than 10 mm Hg respectively. Sixteen (12%) sphygmomanometers were so deteriorated (air leaks, dirt in mercury) that the researcher suggested their immediate withdrawal from service. The results of the blood pressure recording audit suggested digit bias of both systolic and diastolic recordings to the nearest 10 mm Hg. This study suggests that sphygmomanometers used in general practice are very likely to be inaccurate and some may well be so deteriorated that they should be withdrawn from service. The results of the blood pressure audit showed digit bias in systolic and diastolic readings to the nearest 10 mm Hg. The implications for clinical care-both over diagnosis and under diagnosis-although not assessed are likely to be appreciable. PCG Clinical Governance teams in conjunction with Practice Clinical Leads must address these basic issues. PMID- 12082499 TI - Hypertension: a cause of growth impairment. AB - The effects of high blood pressure on growth are not fully understood and while hypertension may be associated with failure to thrive, hypertension causing failure to thrive in children is poorly documented. We describe four children presenting with failure to thrive due to hypertension consequent to various aetiologies. Control of hypertension with appropriate therapy resulted in improved growth. The exact pathogenesis of failure to thrive in hypertensive children is not known. These cases demonstrate the importance of careful measurement of blood pressure in children with failure to thrive. PMID- 12082500 TI - Abdominal aortic aneurysm and renal artery stenosis: renal function and blood pressure before and after endovascular treatment. AB - We describe three patients with abdominal aortic aneurysm (AAA) and renal artery stenosis (RAS). These patients were treated by placement of an aortic endograft and angioplasty or stenting of the renal artery. After the procedure renal function improved or remained stable in two patients and deteriorated slightly in one. Blood pressure was reduced in one hypertensive patient and remained normal in the other two normotensive patients. In conclusion, simultaneous treatment of AAA and RAS with aortic endograft placement and renal artery angioplasty with or without stent, is a safe and effective technique for selected high-risk patients. PMID- 12082501 TI - Augmentation of antiproteinuric effect by combined therapy with angiotensin II receptor blocker plus calcium channel blocker in a hypertensive patient with IgA glomerulonephritis. PMID- 12082502 TI - Succinate dehydrogenase and human diseases: new insights into a well-known enzyme. AB - Inherited defects of the mitochondrial succinate dehydrogenase (SDH) in humans are associated with striking variable clinical presentations ranging from early onset devastating encephalomyopathy to tumour susceptibility in adulthood, or optic atrophy in the elderly. Although different genes encoding the four subunits of the SDH have been found mutated in association with these different phenotypes, we propose that the wide clinical spectrum actually originates from the specific roles of the SDH in the respiratory chain and the mitochondria. In particular, beside its function in the Krebs cycle and the respiratory chain, the specific redox properties of the enzyme could confer to the SDH a specific function in superoxide handling. PMID- 12082503 TI - Genetic analysis of Paraoxonase (PON1) locus reveals an increased frequency of Arg192 allele in centenarians. AB - Human Paraoxonase (PON1) is a High-Density Lipoprotein (HDL)-associated esterase that hydrolyses lipo-peroxides. PON1 has recently attracted attention as a protective factor against oxidative modification of LDL and may therefore play an important role in the prevention of the atherosclerotic process. Two polymorphisms have been extensively studied: a Leucine (L allele) to Methionine (M allele) substitution at codon 55, and a Glutamine (A allele) to Arginine (B allele) substitution at codon 192. We have examined these two aminoacidic changes in 579 people aged 20 to 65 years old, and 308 centenarians. We found that the percentage of carriers of the B allele at codon 192 (B+ individuals) is higher in centenarians than in controls (0.539 vs 0.447), moreover we found that among the B+ individuals, the phenomenon was due to an increase of people carrying M alleles at codon 55 locus. In conclusion, we propose that genetic variability at PON1 locus affects survival at extreme advanced age. PMID- 12082504 TI - Prenatal detection of the 17p11.2 duplication in Charcot-Marie-Tooth disease type 1A: necessity of a multidisciplinary approach for heterogeneous disorders. AB - Charcot-Marie-Tooth (CMT) disease is a typical example of a clinically and genetically heterogeneous disorder and, in most cases, is dominantly inherited and caused by a 1.5 megabase duplication on chromosome 17p11.2 containing the PMP22 gene. This is a non-lethal disease with a wide spectrum of severity, from asymptomatism to severe motor and sensory disability. Unpredictable degree of disability is usually the reason why prenatal diagnosis is required and must be addressed. Molecular procedures such as the use of polymorphic non microsatellite STRs, allowing very fast and reliable results even when requiring a gene dosage interpretation are now available and have been recently validated in post-natal diagnosis. Our results indicate that this approach is also the best-adapted method in case of prenatal diagnosis. Nevertheless, ethical considerations raised by prenatal diagnosis in CMT and more generally in non-lethal disorders remain to be actively considered. Here, we present our experience in genetic counselling, and address the psychological issues for 7 CMT at risk pregnancies. In five cases, a CMT1A duplication was evidenced; pregnancy was terminated in four of these cases and the parents from one affected foetus decided to pursue the pregnancy. PMID- 12082505 TI - Guidelines for the appropriate use of genetic tests in infertile couples. AB - Research on genetic causes of male and female infertility rapidly expanded in the last years, following the development of in vitro fertilising techniques. Genetic tests are now available to explore the cause of the infertility and assess the risk of a given couple to transmit its genetic characteristics. This allows at risk couples to take an informed decision when electing for a medically assisted reproduction. It also allows the professionals to offer a prenatal diagnosis when appropriate. Thus, the genetic work-up of the infertile couple has become good practice for an appropriate diagnosis, treatment and prognostic assessment. The lack of national or international rules for the genetic approach to the infertile couple, prompted the Italian community of professionals in the field of reproductive medicine to join and set up guidelines for the genetic diagnosis of male and female infertility. The group of clinical and research experts is representative of 12 national scientific societies and was supported by external experts from four international societies. We examine the clinically relevant genetic causes of male and female infertility and suggest the category of patients for which each genetic test is recommended or optional, both for an accurate diagnosis and prior to ART. PMID- 12082506 TI - Comparison of family based haplotype methods using intragenic SNPs in candidate genes. AB - The choice of an optimal marker strategy while analysing intragenic SNPs is presently of crucial importance, given the increasing amount of available data. Classical case/control association studies or family based association tests such as the TDT are very popular. However, as these methods are not able to analyse multiple markers simultaneously, different extensions have been proposed in order to use multiple markers. In the present study, the efficiency of five family based haplotypic methods to detect the role of candidate genes is evaluated and compared between them and with the classical single point TDT. Simulations of intragenic SNP maps are performed in recently founded populations. One or several SNPs are assumed to be the functional polymorphisms following different genetic models. Different modes of SNP combinations underlying the genetic susceptibility (epistasis or heterogeneity) are considered. Whereas haplotypic methods perform better in situations of heterogeneity, the TDT remains the most powerful approach in epistasis models as long as the marginal effect of one the SNPs involved in the susceptibility remains important. Haplotypic methods perform better than the TDT when the marginal effect of each SNP is small. Given the similar characteristics of intragenic LD in both old large populations and recently founded populations, in particular the weak correlation between LD and distance, our results are not likely to be specific to founder populations and can be generalized. PMID- 12082507 TI - Foetal haemoglobin in normal healthy adults: relationship with polymorphic sequences cis to the beta globin gene. AB - We evaluated the effects of polymorphic markers within the beta globin gene cluster on HbF expression in two groups. These groups were randomly selected from a survey of HbF distribution in a large population study of unrelated healthy Algerian adults (n=827). The first group contained individuals with normal HbF levels (0.1-0.5%) and the second group contained individuals with raised HbF levels (0.8-2.3%). Of the various polymorphic markers analysed, only the -309 G gamma A-->G, the -158 G gamma C-->T, the G gamma IVS2 TC (TG)(9) AG (TG)(2) (CG)(2) and the -540 beta (AT)(9) T(5) sequence configurations were significantly associated with increased HbF levels. More than 84% of the subjects with elevated HbF levels carried one or several of these four marker configurations, suggesting that the beta globin gene cluster exerts a significant effect on HbF expression in healthy individuals. PMID- 12082508 TI - Combined segregation and linkage analysis of HLA markers in familial psoriasis. AB - Marker-based segregation analysis (MBSA) is a modification of a published method of combined linkage and segregation analysis (Am J Hum Genet 51: 1111-1126, 1992), to determine whether a candidate gene known to be associated with the disease of interest is truly segregating with the disease in families. Here we outline the conceptual basis of MBSA and present a Monte Carlo method for significance testing. The method is applied to PSORS1, a locus within the major histocompatibility complex (MHC) for which linkage and linkage disequilibrium with psoriasis has already been demonstrated. The results are very consistent with our current knowledge of PSORS1, and suggest that MBSA can provide useful information on genotype-phenotype relationships such as penetrance and allelic heterogeneity. PMID- 12082510 TI - Summary--European food consumption survey method. PMID- 12082509 TI - A patient severely affected by spinal neurofibromas carries a recurrent splice site mutation in the NF1 gene. AB - Spinal neurofibromas are found in up to 38% of NF1 patients. However, they cause clinical implications only in about 5% of the patients. In contrast, multiple symptomatic spinal neurofibromas are the main clinical finding in patients with familial spinal neurofibromatosis. Familial spinal neurofibromatosis has been considered to be a distinct clinical form of neurofibromatosis. Linkage analysis in two families and identification of a NF1 gene mutation in a third family strongly associate spinal neurofibromatosis with the NF1 gene. We describe a NF1 patient who satisfies the NIH diagnostic criteria and has severe spinal involvement with bilateral spinal root neurofibromas at every level. A recurrent splice site mutation (IVS19b-3C>G) was identified in the NF1 gene in the patient. We discuss the possibility that the clinical picture of this patient represents an additional example of spinal neurofibromatosis. By comparison of the clinical expression of NF1 in this patient and that in another patient with the identical mutation the hypothesis that spinal neurofibromatosis is associated with a particular mutation is highly unlikely. The involvement of other genes linked to the NF1 gene or modifying genes is currently the most likely explanation for the clinical phenotype of spinal neurofibromatosis. PMID- 12082511 TI - Rationale and methods of the EFCOSUM project. AB - OBJECTIVE: To describe the rationale and methods for a European project (EFCOSUM) to develop a method for a European food consumption survey that delivers internationally comparable data on a set of policy-relevant nutritional indicators. RATIONALE AND METHODS: Currently Member States are collecting data and information for use at national level. At an international level, such data are often of limited comparability and of varying quality. To promote the development and exchange of adequate, reliable and comparable indicators of public health, and the structures needed to exchange the relevant data, a programme of Community action on health monitoring was set up for the EU. The objective of the action programme is to contribute to the establishment of a Community Health Monitoring System. Data will be made available to all Member States via a telematic network, which is currently being developed for the Health Monitoring Programme. With regard to nutrition, there is a need for a limited set of policy-relevant dietary indicators that are comparable among EU Member States. In the field of nutrition, however, there is a regrettable lack of internationally comparable data. The project 'European Food Consumption Survey Method' (EFCOSUM) therefore aimed to define a (minimum) set of dietary components which are relevant determinants of health and to define a method for the monitoring of food consumption in nationally representative samples of all age sex categories in Europe in a comparable way. The project was carried out by 14 Member States as well as nine other European countries. Activities of the project included plenary sessions, desk research, and working group activities, building on existing experience from such projects as DAFNE, EPIC, FLAIR Eurofoods-Enfant project, COST Action 99 and others. The proposed method may be used alone, or as a calibration method to accompany existing ongoing studies. PMID- 12082512 TI - Selection of relevant dietary indicators for health. AB - OBJECTIVE: To define a set of dietary components that are relevant determinants for health in Europe. The selected components are intended to serve as nutrition indicators for health in the European Health Monitoring Programme and, as such, must be limited in number, relevant to health in Europe and practical for all involved countries with respect to data gathering and comparability of data. DESIGN: Major nutrition factors were determined by reviewing relevant epidemiological and clinical literature in nutrition and health as well as referring to reports from international expert groups, including the report from the project Nutrition and Diet for Healthy Lifestyles in Europe. The selection of factors was also based on the relative ease and cost involved for participating countries to obtain comparable and valid data. RESULTS: The selected factors include foods or food groups as well as individual nutrients. Biomarkers are suggested for selected nutrients that pose the greatest difficulty in obtaining valid and comparable data from dietary studies. CONCLUSIONS: The following list of diet indicators for health monitoring in Europe was agreed upon by the EFCOSUM group in 2001, in order of priority: vegetables, fruit, bread, fish, saturated fatty acids as percentage of energy (%E), total fat as %E, and ethanol in grams per day. Biomarkers were suggested for the following nutrients: folate, vitamin D, iron, iodine and sodium. Energy has to be assessed in order to calculate %E from total fat and saturated fatty acids. PMID- 12082513 TI - The use of biomarkers in multicentric studies with particular consideration of iodine, sodium, iron, folate and vitamin D. AB - OBJECTIVE: To determine which biomarkers should be used in a European monitoring system to assess intake of iodine, sodium, iron, folate and vitamin D. These nutrients are relevant for human health but difficult to assess in the diet in a comparable way among countries. RESULTS: Iodine and sodium are mainly excreted in urine. Mean population intakes of iodine and sodium can be estimated by casual urine samples. However, habitual individual intake assessments require multiple 24 h urine collections. Serum ferritin is a sensitive test of iron status (mobilizable storage iron) and continues to be the leading single determination for individual iron status, but with serum transferrin receptor as a promising alternative because it is more specific than ferritin. Erythrocyte folate and homocysteine in blood are markers of folate status which are able to discriminate between normal status and levels of insufficiency in an individual. Serum 25 hydroxyvitamin D is a good marker for vitamin D deficiency in the individual, and can distinguish between overt deficiency and marginal deficiency, especially if combined with serum parathyroid hormone. CONCLUSIONS: Specific and sensitive markers for the dietary intake of iodine, sodium, iron, folate and vitamin D are available, which have the advantage of being more accurate than intake estimations derived from dietary surveys. However, the use of biomarkers also has disadvantages such as extra burden in terms of logistics, budget and ethical approval, and can moreover reduce the participation rate. More basic data for the validity of cross-country comparisons are needed and ideas in terms of study design and sampling frame. PMID- 12082514 TI - Improvement of comparability of dietary intake assessment using currently available individual food consumption surveys. AB - BACKGROUND: Food consumption data are used for monitoring dietary indicators of health. In this context, there is a need for comparable (individual) data at the European level. The preference is to perform a pan-European survey able to generate data collected on the same basis. Until this can be realized, the existing data have to be made as comparable as possible. OBJECTIVE: To identify solutions; to make existing food consumption data from nationally representative databases more comparable. METHODS: General guidelines for the comparison of food intake data using currently available data were discussed and agreed upon in the EFCOSUM project team. RESULTS AND CONCLUSIONS: Criteria were assessed with regard to the population involved, age of the survey, method of data collection, duration of the survey, the food classification system and the food composition tbl Based on these criteria, a maximum of 15 countries could provide food consumption data that can be made comparable at the individual level for the adult population. It is recommended to make data comparable at the food level, starting with vegetables (excluding potatoes), fruits (excluding fruit juices), fish (including shellfish) and bread. Comparability of foods is only possible at the 'raw edible' ingredient level. To achieve this, a large amount of work has to be undertaken. The approach of the EFG (Euro Food Groups) system is considered to be the best compromise between the different classification systems. Comparability at the nutrient level has to wait for the availability of a European Nutrient Database, like the one that is being developed within the EPIC context. PMID- 12082515 TI - Selection of methodology to assess food intake. AB - OBJECTIVE: The aim of the EFCOSUM project was to develop a method to estimate both acute and usual consumption levels in European countries and for the sake of comparison, a common basic method for dietary assessment was needed. The method should allow a reliable comparison of the intake of relevant dietary indicators among large population groups. DESIGN AND RESULTS: The selection of methodology was based on the available literature and the expertise of the participants. To guide the selection process, decision trees containing all relevant moments of choice, alternatives and criteria for the selection of a method for food consumption measurement were developed. Considering that insight into average food and nutrient intake and their distribution in well-defined groups of individuals was important, it was decided that for the monitoring of the selected dietary indicators food consumption data should be collected at an individual level. Different methods were reviewed (24 h recall, dietary record, food frequency questionnaire, dietary history method). CONCLUSIONS: Since the 24 h recall method is applicable in large European populations of different ethnicity, has a relatively low respondent and interviewer burden, is open-ended and is cost effective, this method can be considered as the best method for EFCOSUM to get population mean intakes and distributions for subjects aged 10 y and over in different European countries. Usual intake should be estimated by statistical modelling techniques, using two non-consecutive 24 h recalls and a food list to assess the proportion non-users for infrequently consumed foods. PMID- 12082516 TI - Selection of a food classification system and a food composition database for future food consumption surveys. AB - OBJECTIVE: To harmonize food classification and food composition databases, allowing comparability of consumption at both food and nutrient levels in Europe. DESIGN: To establish the level of comparability at the food level, the EFCOSUM group benefited from the work already carried out within other European projects, which established a Euro Food Groups (EFG) classification system. Four food groups, ie bread, vegetables (excluding potatoes), fruits (excluding fruit juice) and fish and seafood, were judged on their applicability for making food consumption data comparable across countries at the food level. CONCLUSIONS: It was concluded that the EFG system could be used but that still much work has to be done. For food consumption data to be collected in the future, the software that will be used should enable conversion of foods 'as consumed' to foods at the 'raw edible' level. With respect to comparability of nutrient intake estimations, EFCOSUM advises waiting for the European Nutrient Composition Database (ENDB) currently being prepared by the EPIC group. Until this is available, comparison of consumption data at the nutrient level cannot be carried out between countries. PMID- 12082517 TI - Some statistical aspects of food intake assessment. AB - OBJECTIVE: To present the results of the statistical working group of the EFCOSUM project on estimating the minimum sample size for a pan-European dietary survey. BACKGROUND AND METHODS: Numerous statistical issues are involved when planning a nutritional survey aimed at evaluating various indicators, especially if it will be carried out in different countries. The plenary workshop of the EFCOSUM project has chosen four relevant statistical topics: the sample size estimation for dietary surveys, the number of repeated measurements needed to estimate usual intake for each individual; the statistical presentation of data; and the statistical procedures for estimating the usual intake distribution from a limited number of days of observation. This article deals with the first three topics mentioned. The participants of the EFCOSUM project answered a small questionnaire in order to get agreement on the method of estimating a minimum sample size in the context of a monitoring of dietary indicators. Data on the variability of dietary indicators of interest was also collected, in order to calculate a minimum sample size. RESULTS AND CONCLUSION: The main result was that a minimum sample size of 2000 adults in each European country will be needed in order to identify trends in the mean intake of the most relevant foods and nutrients in Europe. This sample size should be higher if trends have to be indentified for socio-demographic subgroups. PMID- 12082518 TI - Estimating the distribution of usual dietary intake by short-term measurements. AB - OBJECTIVE: To estimate the habitual dietary intake distribution in a population on the basis of repeated short-term measurements, especially of multiple 24 h diet recalls. METHODS: Six different statistical methods were evaluated and compared. The comparison referred to theoretical assumptions, admitted data transformations, statistical foundations, available software packages, and applications to real data of dietary intake. RESULTS: The Nusser method and a simplified version of it proposed in the paper have proved to be universally applicable methods for estimating the usual intake distribution for food groups and nutrients. Also, the Buck method seemed to be a robust estimation procedure suitable for the description of food consumption data, whereas the other considered methods were only applicable for log-normally distributed intake data or required a comprehensive data simulation. Characteristics of the estimated usual intake distribution were a decreased standard deviation, increased lower percentiles, and decreased upper percentiles compared to the observed sample distribution of individual means. Empirical results concerning total fat and vegetable intake in three different European consumption surveys showed that the estimated percentiles of the usual intake distribution did not depend markedly on the number of sampling days. CONCLUSIONS: Repeated short-term measurements like 24 h diet recalls can be used to describe the habitual dietary intake distribution in food consumption surveys. Recommended is a sampling design of two non-consecutive sampling days. The sampling days of all participants should be selected in such a way that they cover all seasons and days of the week. PMID- 12082519 TI - Perspectives of using the EPIC-SOFT programme in the context of pan-European nutritional monitoring surveys: methodological and practical implications. AB - OBJECTIVE: To evaluate the usefulness of a computerized 24 h diet recall programme (EPIC-SOFT) for pan-European dietary surveys. METHODS: The perspectives of using EPIC-SOFT in study contexts other than EPIC were evaluated. In order to estimate the usefulness of existing EPIC-SOFT material for developing new versions for non-EPIC countries, a comparison of food and recipe lists and portion pictures between a country participating in EPIC (Sweden, n=2311 foods, 140 photo series) and a non-EPIC country (Finland, FINDIET 1997 Study, n=1373 foods, 126 photo series) was performed. RESULTS: Despite certain limitations, the EPIC-SOFT programme is a promising tool for pan-European Dietary Surveys. About 12-17 man-months would be needed to develop a new version for a non-EPIC country, depending on the extra work required. A 95% coverage of food and mixed recipes was found between the EPIC-Sweden and FINDIET 1997 databases. Of the pictures in the Finnish portion size book, 62% had an equivalent in the EPIC-SOFT picture book, but the range and size of photo series was larger in the latter than in the former, especially for vegetables. A tendency was observed that, if the central serving in a given picture book was larger, the consumers in that country also reported a larger mean portion. CONCLUSIONS: This report shows that the EPIC-SOFT programme is already available for use in 11 European countries and could be extended easily to other countries if resources are made available. However, further developments are required to obtain standardized nutrient estimates and make the maintenance of the EPIC-SOFT databases independent from the EPIC logistics. The available EPIC-SOFT material would be useful for developing new versions for countries of similar food culture. PMID- 12082520 TI - Operationalization of food consumption surveys in Europe: recommendations from the European Food Consumption Survey Methods (EFCOSUM) Project. AB - BACKGROUND: The EFCOSUM (European Food Consumption Survey Methods) Project aims at harmonizing food consumption surveys in European countries within the perspective of an overall Public Health Monitoring Programme. Harmonization implies the need for a common framework of procedures and tools, that are applicable and feasible in all potentially interested countries. A major element in such a framework is the protocol for the operationalization of a food consumption survey, referring to all practical, logistical and material conditions that need to be fulfilled in order to guarantee a successful implementation of such a survey. OBJECTIVE: The objective of this paper was to review a number of aspects of operationalization of food consumption surveys in detail. On the basis of the currently available knowledge and experience in Europe, consensus recommendations have been elaborated for an operationalization protocol that would be feasible for all European countries. METHODS: The EFCOSUM recommendations with respect to operationalization of food consumption surveys in Europe are mainly based on three sources of information, which have been discussed at several ad hoc expert meetings: experience from previous collaborative epidemiological studies, literature searches and results of two questionnaires, distributed among representatives from 23 European countries-all experts affiliated with experienced organizations in the fields of nutrition research and related fields. RESULTS: Consensus recommendations have been set up for the following topics: sampling, recruitment, fieldwork, biomarkers, interviewer qualifications and training and quality control. These recommendations have to be considered the best achievable common denominator within Europe at this time. In a number of cases, recommendations are presented in a hierarchical way, with a gradation from first choice options towards acceptable alternatives. CONCLUSIONS: It can be concluded that a high degree of standardization of fieldwork can be achieved within Europe. A number of specific problems and constraints will have to be solved in connection with the conduction of a real survey. These problems include, amongst others, country-specific decisions on, for example, target population, detailed sampling and recruiting procedures, interview setting and support. However, on the whole, these problems can be overcome and the main recommendations presented in this paper are considered feasible for every country in Europe. Application of the EFCOSUM operationalization protocol in European countries will contribute significantly to the quality and comparability of food consumption data across Europe. It is anticipated that the policy supporting and orienting potential of this type of databases-both at the national and at the EU supranational level-will be increased accordingly. PMID- 12082521 TI - A European food consumption survey method--conclusions and recommendations. AB - OBJECTIVE: To discuss the general outcome and conclusions of a European project (EFCOSUM); to develop a method for a European food consumption survey that delivers internationally comparable data on a set of policy relevant nutritional indicators. DESIGN AND METHODS: The EFCOSUM project was carried out within the framework of the European Health Monitoring Programme by 14 Member States as well as nine other European countries. Activities of the project included plenary sessions, desk research and working group activities, building on existing experience from such projects as DAFNE, EPIC, FLAIR Eurofoods-Enfant project, COST Action 99 and others. All participating states took part in one or more working group activities, which were discussed, adapted and finally approved in plenary sessions. RESULTS AND CONCLUSIONS: For a limited number of countries available food consumption data can be made comparable at the food intake level, but not at the nutrient level. To achieve comparability at the food intake level, a considerable amount of work still has to be done. A minimum list of dietary indicators considered to be the most relevant to be collected for the Health information exchange and monitoring system was identified. As the most suitable method to get internationally comparable new data on population means and distributions of actual intake the 24 h recall was selected, to be conducted at least twice. This also allows for the estimation of usual intake by a modelling technique that separates intra- and inter-individual intake. For a number of micronutrients the use of biomarkers is recommended. Aspects of food classification and food composition were discussed thoroughly, as well as statistical and data collection aspects. For the implementation of a pan-European survey the establishment of a European co-ordinating centre is recommended. The standardization of field work work procedures and other aspects of operationalization have been discussed in detail. It is concluded that there is broad European consensus on the most suitable method for a pan-European dietary survey for the purposes of the EU Health Monitoring Programme. It is further concluded that such a pan-European survey is feasible, if the funds are made available. PMID- 12082524 TI - Activation of ARF by oncogenic stress in mouse fibroblasts is independent of E2F1 and E2F2. AB - The ARF tumour suppressor protein (p14(ARF) in human and p19(ARF) in mouse) is a major mediator of the activation of p53 in response to oncogenic stress. Little is known about the signalling pathways connecting oncogenic stimuli to the activation of ARF. Regulation of ARF occurs primarily at the transcriptional level and several modulators of ARF transcription have been identified. Notably, ectopic expression of E2F1 upregulates ARF transcriptionally, and both E2F1 and ARF have been implicated in apoptosis and cell-cycle arrest. We have used primary mouse fibroblasts deficient for E2F1, E2F2, or both to determine the possible role of these E2F proteins as upstream regulators of ARF in response to oncogenic stimuli and other stresses. In particular, we have studied the effects of oncogenic Ras and the viral oncoprotein E1A on ARF levels, neoplastic transformation, and sensitization to apoptosis. We have also examined the behaviour of the E2F-deficient MEFs with respect to immortalization and sensitivity to DNA damage. None of the ARF-mediated responses that we have analysed is significantly affected in E2F1(-/-), E2F2(-/-) or E2F1/2(-/-) MEFs, and ARF is upregulated normally in all cases. Taken together, our results indicate that the activation of ARF in response to oncogenic stress can occur by E2F1- and E2F2-independent mechanisms. This challenges previous suggestions implicating E2F factors as key mediators in the activation of ARF by oncogenic stress. PMID- 12082525 TI - Wnt-4 regulation by the Wilms' tumour suppressor gene, WT1. AB - The Wilms' tumour suppressor gene, WT1, encodes multiple nuclear protein isoforms, all containing four C-terminal zinc finger motifs. WT1 proteins can both activate and repress putative target genes in vitro, although the in vivo relevance of these putative target genes is often unverified. WT1 mutations can result in Wilms' tumour and the Denys-Drash Syndrome (DDS) of infantile nephropathy, XY pseudohermaphroditism and predisposition to Wilms' tumour. We have established stable transfectants of the mouse mesonephric cell line, M15, which express WT1 harbouring a common DDS point mutation (R394W). A comparison of the expression profiles of M15 and transfectant C2A was performed using Nylon based arrays. Very few genes showed differential expression. However Wnt-4, a member of the Wnt gene family of secreted glycoproteins, was downregulated in C2A and other similar clones. Doxycycline induction of WT1-A or WT1-D expression in HEK293 stable transfectants also elicited an elevation in Wnt4 expression. Wnt4 is critical for the mesenchyme-to-epithelial transition during kidney development, making it an attractive putative WT1 target. We have mapped human Wnt-4 gene to chromosome 1p35-36, a region of frequent LOH in WT, have characterized the genomic structure of the human Wnt-4 gene and isolated 9 kb of immediate promoter. While several potential WT1 binding sites exist within this promoter, reporter analysis does not strongly support the direct regulation of Wnt4 by WT1. We propose that Wnt-4 regulation by WT1 occurs at a more distant promoter or enhancer site, or is indirect. PMID- 12082526 TI - RNA polymerase III transcription can be derepressed by oncogenes or mutations that compromise p53 function in tumours and Li-Fraumeni syndrome. AB - RNA polymerase (pol) III synthesizes essential small RNAs, including tRNA and 5S rRNA. Wild-type p53 can repress pol III transcription both in vitro and in vivo. Many tumours carry substitutions in p53 which have selective effects on its functions. We identify tumour-derived mutations that compromise the ability of p53 to regulate pol III transcription. Furthermore, substitution R175H, the most common mutation in cancers, converts p53 from a repressor to an activator of pol III. Oncoproteins neutralize p53 in some tumours; we show that human papillomavirus E6 and cellular hdm2 can both release pol III from repression by p53. These data suggest that the restraining influence of p53 on pol III will be lost in many tumours. In addition to these features of sporadic cancers, some individuals inherit mutant forms of p53 and consequently suffer from Li-Fraumeni syndrome, showing genetic predisposition to certain malignancies. We find that pol III transcriptional activity is often highly elevated in primary fibroblasts from Li-Fraumeni patients, especially if the germline p53 mutation is followed by loss of the remaining allele. Our data suggest that p53 status can have a profound effect upon pol III transcription and hence on the biosynthetic capacity of cells. PMID- 12082527 TI - Thyroid cell transformation requires the expression of the HMGA1 proteins. AB - Elevated expression of HMGA1 and HMGA2 proteins is correlated with a highly malignant phenotype in several human tumors. We previously demonstrated that the block of HMGA2 protein synthesis prevented rat thyroid cell transformation by murine retroviruses. Suppression of HMGA2 synthesis was associated with lack of induction of HMGA1 proteins suggesting that both HMGA1 and HMGA2 play a role in the process of neoplastic transformation. To determine the role of the HMGA1 gene in thyroid cell transformation, we blocked HMGA1 protein synthesis by an antisense methodology. Here we report that transfection of an HMGA1 cDNA antisense construct into a normal rat thyroid cell line (FRTL-5 Cl2), followed by infection with Kirsten murine sarcoma virus (KiMSV), generated a transformed cell line that expresses high levels of the v-ras-Ki oncogene and that does not require thyroid-stimulating hormones for growth. However, this cell line does not show the malignant phenotype, i.e., it neither grows in soft agar nor induces tumors after injection in athymic mice. Moreover, the lack of the neoplastic phenotype in the virus-infected thyroid cells carrying the HMGA1 antisense construct correlates with the absence of induction of AP-1 transcriptional activity. PMID- 12082528 TI - c-MYC overexpression in Ba/F3 cells simultaneously elicits genomic instability and apoptosis. AB - Overexpression of c-Myc in tumors is usually associated with cell proliferation and increased susceptibility to apoptosis. Concomitantly, c-Myc contributes to tumorigenesis by its ability to destabilize the cellular genome. Here, we examined whether c-Myc induces genomic instability and apoptosis in c-Myc activated cells. Wild-type Myc (wt-Myc) and two mutated Myc myc box II proteins (mt-Myc) were overexpressed in IL3-dependent murine Ba/F3 cells. As expected, wt Myc triggered apoptosis in absence of IL3. Standard karyotyping, spectral karyotyping, and fluorescent in situ hybridization (FISH) were performed before and after c-Myc activation. Structural and numerical genomic instability was detected 48 h after wt-Myc activation and included gene amplification, the formation of extrachromosomal elements (EEs), chromosome breakage, deletions, increased aneuploidy, and polyploidization. Interestingly, some cells simultaneously displayed genomic instability and apoptosis. Both wt- and mt-Myc proteins were equally potent promoters of genomic instability. However, only wt Myc simultaneously induced genomic instability and apoptosis. Mt-Myc proteins failed to induce apoptosis, thereby generating a strong imbalance towards the survival of genomically unstable cells. PMID- 12082529 TI - Regulation of apoptosis by p53 in UV-irradiated human epidermis, psoriatic plaques and senescent keratinocytes. AB - The carcinogenic effects of sunlight in human epidermis may be thwarted by either: transient growth arrest and repair of DNA photodamage in keratinocytes (KCs); elimination of KCs with damaged DNA via apoptosis; or by stimulating a senescence switch whereby KCs become irreversibly growth arrested. Using normal human skin organ cultures and living epidermal equivalents, we demonstrate that in the proliferative basal layer, removal of KCs via apoptosis had a rapid onset (beginning within 2 h) following UV-light exposure generating progressively greater numbers of KCs with thymine dimers as the dose of UV-light was increased; involved induction of Apaf-1, activation of caspase-3, and was dependent on p53 activation as addition of a p53 chemical inhibitor blocked the apoptotic response. Suprabasal layer KCs underwent apoptosis at much later time points (>8 h). KCs in the basal layer repaired DNA damage more rapidly than KCs in suprabasal layers. Steady state levels of p53 increased in irradiated cells, and the increase was accompanied by phosphorylation of serine 9 and serine 15, but not serine 6 residues. By contrast, cultured KCs undergoing spontaneous replicative senescence were resistant to UV-induced apoptosis. Senescent KCs constitutively contained low levels of p53, which were neither increased nor phosphorylated or acetylated after UV-exposure and possessed minimal DNA binding activity, indicative of functional inactivation. Furthermore, treatment of senescent KCs with DNA damaging agent adriamycin did not result in activation of latent p53 or apoptosis. When KCs within psoriatic plaques were examined, they resembled senescent KCs in that they expressed p53, which was not phosphorylated or acetylated. Thus, UV-light induces DNA damage in human epidermal KCs triggering p53 activation, and subsequent apoptosis involving distinct cell layers and kinetics. However, the lack of p53 activation as seen in senescent KCs and psoriatic plaques, is associated with a relative resistance of KCs to UV induced apoptosis. In conclusion, the sensitivity and resistance of KCs to apoptosis depends not only on the location within various layers of epidermis and levels of p53, but may also involve p53 activation via post-translational modifications. PMID- 12082530 TI - Interaction and colocalization of PGP9.5 with JAB1 and p27(Kip1). AB - PGP9.5 (UCH-L1) is a member of the ubiquitin C-terminal hydrolase (UCH) family of proteins that is expressed in neuronal tissues. Our previous studies have shown that PGP9.5 was highly expressed in primary lung cancers and lung cancer cell lines. Additionally, the frequency of PGP9.5 over expression increases with tumor stage, indicating that PGP9.5 may play a role in lung cancer tumorigenesis. We used the yeast two-hybrid system to identify proteins that interact with PGP9.5. We show that PGP9.5 interacts with at least three proteins, one of which is JAB1, a Jun activation domain binding protein that can bind to p27(Kip1) and is involved in the cytoplasmic transportation of p27(Kip1) for its degradation. We also show that PGP9.5 is associated with JAB1 in vitro and in vivo; and that both proteins can be a part of a heteromeric complex containing p27(Kip1) in the nucleus in lung cancer cells. Furthermore, under serum-restimulation, nuclear translocation of both PGP9.5 and JAB1 coincides with a reduced level of p27(Kip1) in the nucleus. In contrast, when cells are contact inhibited, both PGP9.5 and JAB1 became more perinuclear and cytoplasmic in localization while p27(Kip1) was present only in the nucleus. Therefore, PGP9.5 may contribute to p27(Kip1) degradation via its interaction and nuclear translocation with JAB1. PMID- 12082531 TI - Reconstitution of hTERT restores tumorigenicity in melanoma-derived c-Myc low expressing clones. AB - c-Myc is involved in the control of telomerase activity through its ability to induce the expression of the catalytic subunit of the enzyme, the human telomerase reverse transcriptase (hTERT). Our aim was to study whether telomerase plays a critical role in c-Myc-dependent tumorigenicity of melanoma cells. By using M14-derived clones, expressing low levels of c-Myc, we demonstrated that the down-regulation of c-Myc reduced cell proliferation rate, cloning efficiency and tumorigenicity and increased the apoptotic rate. Decreased tumorigenic potential correlated with reduced hTERT gene expression, telomerase activity and telomere shortening. Introduction of wild-type hTERT into these cells increased their proliferation rate and partially re-established their tumorigenic potential, at early passages, even though the apoptotic rate of the population remained unaltered. After several in vitro passages, hTERT-mediated cell proliferation made the tumorigenic potential of the c-Myc low-expressing clones comparable to that of the M14 parental line. Over-expression of the mutant biologically inactive hTERT did not drive cells to proliferate. In conclusion, our results demonstrate that the reconstitution of high levels of telomerase activity reverses the low tumorigenicity due to low c-Myc expression. PMID- 12082532 TI - Tumour specific promoter region methylation of the human homologue of the Drosophila Roundabout gene DUTT1 (ROBO1) in human cancers. AB - The human homologue of the Drosophila Roundabout gene DUTT1 (Deleted in U Twenty Twenty) or ROBO1 (Locus Link ID 6091), a member of the NCAM family of receptors, was recently cloned from the lung cancer tumour suppressor gene region 2 (LCTSGR2 or U2020 region) at 3p12. DUTT1 maps within a region of overlapping homozygous deletions characterized in both small cell lung cancer lines (SCLC) and in a breast cancer line. In this report we (a) defined the genomic organization of the DUTT1 gene, (b) performed mutation and expression analysis of DUTT1 in lung, breast and kidney cancers, (c) identified tumour specific promoter region methylation of DUTT1 in human cancers. The gene was found to contain 29 exons and spans at least 240 kb of genomic sequence. The 5' region contains a CpG island, and the poly(A)(+) tail has an atypical 5'-GATAAA-3' signal. We analysed DUTT1 for mutations in lung, breast and kidney cancers, no inactivating mutations were detected by PCR-SSCP. However, seven germline missense changes were found and characterized. DUTT1 expression was not detectable in one out of 18 breast tumour lines analysed by RT-PCR. Bisulfite sequencing of the promoter region of DUTT1 gene in the HTB-19 breast tumour cell line (not expressing DUTT1) showed complete hypermethylation of CpG sites within the promoter region of the DUTT1 gene (-244 to +27 relative to the translation start site). The expression of DUTT1 gene was reactivated in HTB-19 after treatment with the demethylating agent 5-aza-2' deoxycytidine. The same region was also found to be hypermethylated in six out of 32 (19%) primary invasive breast carcinomas and eight out of 44 (18%) primary clear cell renal cell carcinomas (CC-RCC) and in one out of 26 (4%) primary NSCLC tumours. Furthermore 80% of breast and 75% of CC-RCC tumours showing DUTT1 methylation had allelic losses for 3p12 markers hence obeying Knudson's two hit hypothesis. Our findings suggest that DUTT1 warrants further analysis as a candidate for the tumour suppressor gene (TSG) at 3p12, a region defined by hemi and homozygous deletions and functional analysis. PMID- 12082533 TI - Identification of a novel gene NCRMS on chromosome 12q21 with differential expression between rhabdomyosarcoma subtypes. AB - Rhabdomyosarcoma (RMS) is a malignant soft tissue tumor showing varying degrees of skeletal muscle differentiation. Two major histologic subtypes exist, alveolar and embryonal, each with associated molecular genetic changes. We have used Representational Difference Analysis (RDA) to compare gene expression between the two RMS subtypes and have identified the novel gene NCRMS (non-coding RNA in RMS) that has increased expression in the alveolar subtype relative to the embryonal subtype. Multiple alternatively spliced forms of NCRMS were identified through library screening, RACE, and comparison to human expressed sequence tags (ESTs). Northern blot analysis indicated the transcript size to be 1.25 kb in alveolar RMS. There was no sequence homology to any of the known genes in GenBank, but extensive homology to ESTs from various species. Comparison to human genomic sequences identified at least 11 exons mapping to chromosomal region 12q21. Differential expression of NCRMS was noted between various tumor types. Since NCRMS RNA possesses limited potential for protein coding, yet with conserved sequences between different species, it is likely that NCRMS is a functional non coding RNA. Known genes in its proximity include myogenic regulators Myf5 and Myf6, growth factor Igf1, and another potential differentially expressed gene (ATP2B1) in RMS isolated by RDA. PMID- 12082534 TI - Frameshift mutations of RIZ, but no point mutations in RIZ1 exons in malignant melanomas with deletions in 1p36. AB - Recently, the retinoblastoma protein interacting zinc finger gene RIZ has been proposed as a candidate for the tumor suppressor locus on 1p36, because of the common loss of RIZ1 RNA in human tumors. In addition, frameshift mutations of this gene have been demonstrated in a variety of tumors with microsatellite instability. Since alterations in this region have been described in malignant melanomas, we investigated DNA of paraffin-embedded sections from 16 typical naevi, 19 atypical naevi, 33 primary melanoma lesions and 25 metastases and DNA from four melanoma cell lines by PCR and direct sequencing analysis of RIZ. Frameshift mutations in the RIZ gene were found in 17% of melanoma samples and 8.6% of naevi, but we could not demonstrate any missense mutations in the exons of RIZ1. No LOH of the RIZ gene nor any microsatellite instability in six dinucleotide markers or in the mononucleotide repeats IGRIIR, hMSH3, and hMSH6 could be demonstrated in the samples with RIZ frameshift mutations. Although our results do not explain the high rate of deletions in 1p36 found in this tumor, they assign RIZ a potential role in the multi-step tumor forming process of malignant melanoma of the skin. PMID- 12082535 TI - p53 mutations and tetraploids under r- and K-selection. AB - Cotransfection of rat embryo fibroblasts with c-myc and activated H-ras oncogenes is one experimental model of the multistep oncogenesis associated with p53 mutations and aneuploidy. Using the model, we found that selection processes, e.g., r- and K-selection, affect emergence of p53 mutants and tetraploids. Culture optimum for logarithmic growth (r-selection) selected p53 mutants as they proliferated rapidly, while in confluent culture (K-selection) tetraploids emerged regardless of the p53 status. Transfection of the mutated p53 gene with dominant negative functions eradicated untransfected cells under both r- and K selection. However, these p53 mutants can be eradicated under K-selection by cells with normal p53 function and that had been selected under prolonged K selection. The presence of competitors and the type of selection should determine whether or not p53 mutants and/or tetraploids predominate. These observations strengthen the importance of selection processes in case of cancer. PMID- 12082536 TI - Mouse DDA3 gene is a direct transcriptional target of p53 and p73. AB - The p53 tumor suppressor is a transcription factor that activates the expression of many target genes. We have previously reported the identification of a p53 regulated mouse gene DDA3. The 5' upstream genomic region of the mouse DDA3 was cloned, and sequence analysis revealed the presence of a potential p53 response element (RE2) residing at nucleotides +390 approximately +409 relative to the first translation start site. When fused upstream to a luciferase reporter gene, 5' genomic regions of the DDA3 gene containing RE2 were shown to be responsive to the wild-type, but not mutant p53, in a transient transfection assay. RE2 was sufficient to confer the transactivation responsiveness to p53. Furthermore, gel mobility shift analysis showed that RE2 formed specific complexes with wild-type p53. Induction of DDA3 was found in adriamycin treated normal mouse embryonic fibroblast cells (MEF), but not in p53 knockout (p53(-/-)) MEF. Overexpression of p73 induced DDA3 mRNA expression, and luciferase reporter analysis indicated that RE2 was responsive to transactivation by members of the p73 family proteins. Consistent with these findings, elevated expression of p73 protein and DDA3 mRNA were observed concomitantly in the p53(-/-) MEF cells treated with cisplatin. These results together demonstrated that DDA3 is a transcriptional target gene of p53 and its related-protein p73. PMID- 12082537 TI - Ras-inducible immortalized fibroblasts: focus formation without cell cycle deregulation. AB - The Ras oncogene transforms cultured murine fibroblasts into malignant, focus forming cells, whose lack of contact inhibition is evidenced by high saturation densities. In order to investigate the reversibility of Ras transformation, as well as the kinetics of Ras-induced changes, cell lines that conditionally express oncogenic Ras were constructed. Both focus formation and increased saturation density were inducible and fully reversible. In exponentially growing cells, oncogenic Ras-expression had no effect on proliferation rates, Erk phosphorylation, or the level of cyclin D1, and Ras-induction did not confer serum-independent growth. As expected, growth to high density in uninduced cells led to quiescence with a low level of cyclin D1 and no active Erk; in this setting, Ras induction prevented full downregulation of cyclin D1 and inactivation of Erk. Our results show that Ras expression to a level sufficient for transformation leads to relatively subtle effects on known downstream targets, and that the focus formation and increased saturation density growth induced by Ras is not a result of growth factor independence. PMID- 12082538 TI - BCR-ABL alters the proliferation and differentiation response of multipotent hematopoietic cells to stem cell factor. AB - Chronic myeloid leukaemia (CML), a hematopoietic stem cell disorder is characterized by the expression of BCR-ABL. To investigate the effects of BCR-ABL on multipotent hematopoietic cells, a temperature sensitive BCR-ABL tyrosine kinase was expressed in the cell line, FDCP-Mix. BCR-ABL mediated an increase in c-kit expression that correlated with an enhanced mitogenic response to SCF. This was not observed in the absence of Bcr-Abl kinase activity or presence of the BCR ABL inhibitor STI571, which also inhibits c-kit. When cultured in a combination of SCF plus G-CSF the FDCP-Mix cells undergo neutrophilic differentiation over a 7-10 day period. When BCR-ABL was active there was a marked inhibition of cell maturation compared to control cells in which BCR-ABL was either inactive or not present. However, BCR-ABL did not block differentiation as the cells eventually undergo terminal maturation. These data argue that BCR-ABL is directly responsible for the enhanced response to SCF reported in CML progenitor cells. Furthermore, although the primary effect of STI571 is via direct inhibition of BCR-ABL, STI571 additionally reduces the enhanced response to SCF. Thus there are two sites of STI571 action of potential importance in Bcr-Abl expressing cells. PMID- 12082539 TI - Targeted disruption of c-myb in the chicken pre B-cell line DT40. AB - The c-myb proto-oncogene is highly expressed in a wide variety of immature hematopoietic cells and plays a key role in the development of the hematopoietic system. c-myb and its retroviral counterpart v-myb encode transcription factors which have been implicated in the regulation of certain target genes. Targeting of c-myb in mouse embryonic stem cells by homologous recombination has provided clear evidence that c-myb is necessary for the proper development of most myeloid lineages of the hematopoietic system as well as of T-lymphocytes. Here we have explored the function of c-myb in the B-lymphoid lineage. We have used the chicken DT40 cells, a pre B-cell line which shows extremely high efficiencies of homologous recombination, as a model system to disrupt c-myb. DT40 cells lacking a functional c-myb gene are viable and show only minor perturbations of their growth parameters, indicating that c-myb is not an essential gene in these cells. We have used the c-myb null DT40 cells to analyse the expression of genes which have been previously been identified as myb target genes. Neither c-myc nor bcl 2, two putative myb targets, showed altered expression in the cells lacking c myb. However, expression of the Pdcd4 gene, a myb target gene originally identified in a myelomonocytic cell line expressing a conditional form of v-myb, was diminished in the absence of c-myb. The c-myb knock-out cells described here should provide a useful model system for the identification and characterization of c-myb target genes in B-lymphoid cells. PMID- 12082540 TI - p53 regulates Stat3 phosphorylation and DNA binding activity in human prostate cancer cells expressing constitutively active Stat3. AB - Constitutive activation of the signal transducer and activator of transcription 3 (Stat3) and mutation of the p53 are both commonly detected in human prostate cancer cells. We sought to investigate whether there is functional regulation of Stat3 by wild-type (wt) p53. Our results demonstrate that expression of wt p53 but not mutant p53 significantly reduced tyrosine phosphorylation of Stat3 and inhibited Stat3 DNA binding activity in both DU145 and Tsu prostate cancer cell lines that express constitutively active Stat3. Expression of the p53 downstream target, p21(WAF-1), did not have any inhibitory effect on Stat3 phosphorylation. Wt p53 but not p21(WAF-1) induced dramatic apoptosis in these prostate cancer cells. Expression of wt p53 did not cause a reduction of phosphorylation independent Stat3 protein and reduction of phosphorylation of three unrelated protein kinases, ERK1, ERK2 (ERK1/2), and AKT. Interestingly, p53-dependent apoptosis occurred in the presence of high levels of phosphorylated AKT and ERK1/2 in both DU145 and Tsu prostate cancer cells. Further, we evaluated a series of established human prostate, breast, and ovarian cancer cell lines and found that all cancer cell lines expressing constitutively active Stat3, only harbor mutated or deleted p53. One implication of these results is that the anti proliferative activities of p53 may not be compatible with the constitutive Stat3 signal in cancer cells. PMID- 12082541 TI - Genomic organization of a novel glycosylphosphatidylinositol MAM gene expressed in human tissues and tumors. AB - We report the genomic organization of a novel human gene mapped to chromosome 6p21, encoding a putative glycosylphosphatidylinositol (GPI) anchored protein containing a MAM (meprin, A5 antigen, protein tyrosine phosphatase mu) domain, that we have termed as GPIM (GPI and MAM) protein. GPIM gene consists of an 8.9 kb transcript composed of 17 coding exons spanning about 65.5 kb of genomic DNA. The deduced polypeptide consists of 955 amino acids and exhibits structural features found in different types of cell adhesion molecules (CAMs), such as the presence of immunoglobulin domains, the presence of a MAM domain or the capacity to anchor to the cell membrane by a GPI motif. Expression analysis in normal human tissues revealed that this gene is expressed as a 5 kb and 9.5 kb mRNA. Furthermore, the smaller transcript is highly expressed in some human cancer cell lines, as well as in different primary tumors (lung, colon, uterus, stomach and breast). Interestingly, the gene was higher expressed in several tumor tissues analysed as compared to their corresponding normal tissues. Thus, GPIM is a novel gene codifying a protein with structural features characteristics of some CAMs, which might be involved in the tumor progression. PMID- 12082542 TI - Gene expression profiling defines molecular subtypes of classical Hodgkin's disease. AB - Although the prognosis of Hodgkin's disease is relatively good, around 20% of patients do not benefit from current therapies and succumb to their disease. A large-scale molecular characterization of disease might help improve HD management. Using cDNA arrays, we studied the mRNA expression levels of approximately 1000 selected genes in 34 benign and malignant lymphoid samples including 21 classical Hodgkin's disease (HD) tissue samples. Hierarchical clustering identified three main molecular groups of HD tumours relevant with respect to histology and clinical outcome (response to therapy and survival). Samples from all bad outcome HD (BOHD) patients clustered in one group whereas the two other groups contained most good outcome HD (GOHD) cases. The nodular sclerosis GOHD samples overexpressed genes involved in apoptotic induction and cell signalling, including cytokines, while the BOHD samples were characterized by the upregulation of genes involved in fibroblast activation, angiogenesis, extracellular matrix remodelling, cell proliferation, and the downregulation of tumour suppressor genes. Our results establish a molecular taxonomy of HD correlating with response to therapy and clinical outcome, thereby suggesting the possibility of improving the current prognostic classification. PMID- 12082544 TI - MIM, a potential metastasis suppressor gene in bladder cancer. AB - Using a modified version of the mRNA differential display technique, five human bladder cancer cell lines from low grade to metastatic were analyzed to identify differences in gene expression. A 316-bp cDNA (C11-300) was isolated that was not expressed in the metastatic cell line TccSuP. Sequence analysis revealed that this gene was identical to KIAA 0429, has a 5.3-kb transcript that mapped to 8q24.1. The protein is predicted to be 356 amino acids in size and has an actin binding WH2 domain. Northern blot revealed expression in multiple normal tissues, but none in a metastatic breast cancer cell line (SKBR3) or in metastatic prostatic cancer cell lines (LNCaP, PC3). We have named this gene Missing in Metastasis (MIM) and our data suggest that it may be involved in cytoskeletal organization. PMID- 12082543 TI - Mouse models of neurofibromatosis 1 and 2. AB - The neurofibromatoses represent two of the most common inherited tumor predisposition syndromes affecting the nervous system. Individuals with neurofibromatosis 1 (NF1) are prone to the development of astrocytomas and peripheral nerve sheath tumors whereas those affected with neurofibromatosis 2 (NF2) develop schwannomas and meningiomas. The development of traditional homozygous knockout mice has provided insights into the roles of the NF1 and NF2 genes during development and in differentiation, but has been less instructive regarding the contribution of NF1 and NF2 dysfunction to the pathogenesis of specific benign and malignant tumors. Recent progress employing novel mouse targeting strategies has begun to illuminate the roles of the NF1 and NF2 gene products in the molecular pathogenesis of NF-associated tumors. PMID- 12082545 TI - Classification of sensitivity or resistance of cervical cancers to ionizing radiation according to expression profiles of 62 genes selected by cDNA microarray analysis. AB - To identify a set of genes related to radiosensitivity of cervical squamous cell carcinomas and to establish a predictive method, we compared expression profiles of 9 radiosensitive and 10 radioresistant tumors obtained by biopsy before treatment, on a cDNA microarray consisting of 23,040 human genes. We identified 121 genes whose expression was significantly greater in radiosensitive cells than in radioresistant cells, and 50 genes that showed higher levels of expression in radioresistant cells than in radiosensitive cells. Some of these genes had already known to be associated with the radiation response, such as aldehyde dehydrogenase 1 (ALDH1) and X-ray repair cross-complementing 5 (XRCC5) (P<.05, Mann-Whitney test). The validity of the total of 171 genes as radiosensitivity related genes were certified by permutation test (P<.05). Furthermore, we selected 62 genes on the basis of a clustering analysis, and confirmed the validity of these genes with cross-validation test. The cross-validation test also indicates the possibility of making prediction of radiosensitivity for discriminating radiation-sensitive from radiation resistant biopsy samples by predicting score (PS) values calculated from expression values of 62 genes in 19 samples, because the prediction successfully and unequivocally discriminated the radiosensitive phenotype from the radioresistant phenotype in our test panel of 19 cervical carcinomas. The extensive list of genes identified in these experiments provides a large body of potentially valuable information for studying the mechanism(s) of radiosensitivity, and selected 62 genes opens the possibility of providing appropriate and effective radiotherapy to cancer patients. PMID- 12082546 TI - The treatment of malignant meningioma with verotoxin. AB - Malignant meningiomas (MMs) are aggressive intracranial neoplasms with a 75% 5 year recurrence rate. Verotoxin 1 (VT1) is an Escherichia coli toxin, which has recently been shown to have anti-neoplastic action by targeting the globotriosylceramide (Gb(3)) glycolipid on tumor cells and tumor neovasculature. To investigate the potential use of VT1 as a clinical agent for MM, we initially tested 16 meningiomas for Gb(3) expression. Nine of 11 MMs (82%), but only one of five benign meningiomas (20%), were positive for Gb(3). An orthotopic xenograft model was used to test the efficacy of VT1 treatment for MM. We first demonstrated that Gb(3) was highly expressed by the MM cell line, IOMM-Lee, and that this cell line was highly sensitive to VT1 treatment in vitro. A single intratumoral injection of VT1 significantly improved survival in nude mice harboring intracranial tumours (P<.0001). Factor-eight immunostaining of tumours harvested from VT1-treated animals revealed a marked reduction in the tumour microvascular density. In addition, the tumors of VT1-treated animals displayed increased apoptosis by TUNEL analysis and showed a significant decrease in cell proliferation, as determined by MIB-5 immunostaining. VT1 treatment of MM is effective in our orthotopic xenograft model, and warrants further exploration as a potential treatment for these highly anaplastic and aggressive neoplasms. PMID- 12082547 TI - Antitumor activity and prolonged expression from a TRAIL-expressing adenoviral vector. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in a variety of transformed cell lines, but generally spares most normal cells. Transduction by an adenoviral vector expressing human TRAIL cDNA (Ad.TRAIL-GFP) resulted in both direct tumor cell killing as well as a potent bystander effect through presentation of TRAIL by transduced normal cells. Administration of Ad.TRAIL-GFP significantly prolonged survival of mice harboring either intracerebral glioblastomas or breast carcinoma-induced peritoneal carcinomatosis. Additionally, TRAIL induced prolonged transgene expression in normal tissue, presumably as a result of diminished immune-mediated destruction of vector-transduced cells. Taken together, these data suggest that vector mediated transduction of TRAIL may represent an effective strategy for cancer gene therapy. PMID- 12082548 TI - STAT6 mediates interleukin-4 growth inhibition in human breast cancer cells. AB - In addition to acting as a hematopoietic growth factor, interleukin-4 (IL-4) inhibits growth of some transformed cells in vitro and in vivo. In this study, we show that insulin receptor substrate (IRS)-1, IRS-2, and signal transducer and activator of transcription 6 (STAT6) are phosphorylated following IL-4 treatment in MCF-7 breast cancer cells. STAT6 DNA binding is enhanced by IL-4 treatment. STAT6 activation occurs even after IRS-1 depletion, suggesting the two pathways are independent. To examine the role of STAT6 in IL-4-mediated growth inhibition and apoptosis, a full-length STAT6 cDNA was transfected into MCF-7 cells. Transient overexpression of STAT6 resulted in both cytoplasmic and nuclear expression of the protein, increased DNA binding in response to IL-4, and increased transactivation of an IL-4 responsive promoter. In STAT6-transfected cells, basal proliferation was reduced whereas apoptosis was increased. Finally, stable expression of STAT6 resulted in reduced foci formation compared to vector transfected cells alone. These results suggest STAT6 is required for IL-4 mediated growth inhibition and induction of apoptosis in human breast cancer cells. PMID- 12082549 TI - Differential activity of NO synthase inhibitors as chemopreventive agents in a primary rat tracheal epithelial cell transformation system. AB - A model to study the effectiveness of potential chemopreventive agents that inhibit neoplastic process by different mechanisms has been used to test the efficacy of seven nitric oxide synthase (NOS) inhibitors. Five selective inducible NOS (iNOS) inhibitors: S-methyl isothiourea (S-MITU), S-2-aminoethyl isothiourea (S-2-AEITU), S-ethyl isothiourea (S-EITU), aminoguanidine (AG), 2 amino-4-methyl pyridine (2-AMP), and two non selective general NOS inhibitors: l N(6)-(1-iminoethyl) lysine (IEL) and N(omega)-nitro-l-arginine (NNLA), were tested for efficacy against a carcinogen, benzo[a]pyrene (B[a]P)-induced primary rat tracheal epithelial (RTE) cell transformation assay. RTE cells were treated with B[a]P alone or with five nontoxic concentrations of an NOS inhibitor and the resulting foci at the end of 30 days were scored for inhibition of transformation. The results indicate that all three isothiourea compounds inhibited B[a]P-induced RTE foci in a dose-dependent manner. S-AEITU was the most effective inhibitor with an IC(50) (the molar concentration that inhibits transformation by 50%) of 9.1 microM and 100% inhibition at the highest dose tested (30 microM). However, both S-EITU and S-MITU showed a maximum percent inhibition of 81% and 100% at 1 mM with an IC(50) of 84 and 110 microM, respectively. 2-AMP did not show any dose-dependent response, but was highly effective (57% inhibition) at an intermediate dose of 30 microM and an IC(50) of 25 microM. Similar to thiourea compounds, AG exhibited good dose-dependent inhibition with a maximum inhibition of 86% at 1 mM. NNLA and IEL were negative in this assay. Based on the IC(50) values, NOS inhibitors were rated for efficacy from high to low as follows: S-2-AEITU<2-AMP 0.7) linkage disequilibrium (LD) between SNPs was observed across the same 390 kb region associated with the CYP2D6 phenotype. The observed phenotype:genotype association reached genome-wide levels of significance, and supports the strategy for potential application of LD mapping and whole genome association scans to pharmacogenetic studies. PMID- 12082589 TI - Obsessive-Compulsive Disorder, 5-HTTLPR polymorphism and treatment response. AB - Recently, a role for a functional polymorphism within the promoter region of the serotonin transporter gene (5-HTTLPR) in conferring susceptibility to Obsessive Compulsive Disorder (OCD) has been suggested. The aim of this study was to test the hypothesis that allelic variation of the 5-HTTLPR could be associated with OCD susceptibility or influence the drug response in OCD. One hundred and eighty one OCD patients were recruited; 92 patients underwent a standardized treatment with fluvoxamine. No significant differences in allele/genotype distribution of the 5-HTTLPR were found between 191 controls and OCD. No differences in fluvoxamine response in the three genotypes groups in OCD were found, considering Yale-Brown Obsessive Compulsive Scale (YBOCS) total scores. Nevertheless, a significant time per genotype interaction was found for the YBOCS subtotal compulsion scores. Considering patients without tic disorder co-diagnosis, a significant time per genotype interaction for both YBOCS total scores and compulsion scores was found. PMID- 12082590 TI - The contribution of genetic factors to thrombotic and bleeding outcomes in coronary patients randomised to IIb/IIIa antagonists. AB - Genetic variants are risk factors for coronary disease, but their role in recurrent events and in response to treatment is less clear. We genotyped genetic variants implicated in primary coronary disease in 924 Caucasians with acute coronary syndromes participating in the OPUS-TIMI16 trial of the GPIIb/IIIa antagonist orbofiban. These were the platelet glycoprotein (GP) receptors GPIIIa, GPIa, GPIbalpha; platelet ligands beta-fibrinogen and von Willebrand Factor (vWF); interleukins (IL) IL-1RN, and IL-6; adhesion proteins E-selectin and P selectin; and metalloproteinase MMP-9. Cox modelling of all genetic variants demonstrated no significant impact on the composite endpoint (P = 0.88), which included myocardial infarction (MI), death, recurrent ischemia, urgent revascularisation and stroke, but a significant impact on recurrent myocardial infarction alone (chi(2) = 20.4, 10 df, P = 0.04). There was a significant interaction of the polymorphisms with orbofiban treatment influencing bleeding outcomes (P = 0.004). Thus, genetic polymorphisms may be associated with subsequent myocardial infarction, and may also be associated with treatment associated bleeding among coronary patients. PMID- 12082591 TI - A common P-glycoprotein polymorphism is associated with nortriptyline-induced postural hypotension in patients treated for major depression. AB - The multi-drug resistance gene ABCB1 (or MDR1) encodes a P-glycoprotein (P-gp) that regulates passage of many substances across the blood-brain barrier. The antidepressant amitriptyline and its metabolites (including nortriptyline) are substrates for P-gp, and in mice lacking P-gp, penetration of amitriptyline, but not fluoxetine, into the brain is enhanced. We reasoned that polymorphic variation of P-gp may contribute to differing responses of patients to antidepressant drugs. A single nucleotide polymorphism (SNP) of ABCB1 (3435C>T) was recently correlated with expression levels and in vivo function of P-gp. We examined this SNP in patients with major depression enrolled in a randomized antidepressant treatment trial of nortriptyline and fluoxetine, and observed a significant association between nortriptyline-induced postural hypotension and 3435C>T (chi(2) = 6.78, df = 2, P = 0.034). Our results suggest that homozygosity for 3435T alleles of ABCB1 is a risk factor for occurrence of nortriptyline induced postural hypotension (OR = 1.37, P = 0.042, 95% CI 1.01-1.86). PMID- 12082594 TI - Health protection--a strategy and a national agency. AB - The Chief Medical Officer for England has published a strategy for health protection and announced the formation of a new body, the Health Protection Agency, to deal with the threats to health from infectious disease, chemicals, toxins and radiation hazards. This reflects international recognition of the need to combat threats to health from the likes of tuberculosis, HIV, influenza, anti microbial resistance, chemical accidents and bio-terrorism, and the risks to health associated with increased movements of people, animals and goods, climate change and industrialisation. The strategy will strengthen surveillance and response linking contributions from clinical specialities with public health, microbiology, toxicology and radiation science within the health protection family. The Agency will be formed by combining a number of national and specialist public bodies and personnel delivering local protection services. The strategy represents a unique opportunity to strengthen local and national structures and develop a world-class health protection service. Detailed plans are being developed by the Department of Health with a variety of stakeholders for the launch of the Agency in 2003. A number of challenges will have to be met including developing and training the workforce in health protection, providing career structures for public health scientists and nurses, strengthening electronic communications and developing health protection networks within broader public health structures. Health protection should now be recognised as a sub-speciality of public health. PMID- 12082592 TI - Multi-locus interactions predict risk for post-PTCA restenosis: an approach to the genetic analysis of common complex disease. AB - The complexity of recognizing the potential contribution of a number of possible predictors of complex disorders is increasingly challenging with the application of large-scale single nucleotide polymorphism (SNP) typing. In the search for putative genetic factors predisposing to coronary artery restenosis following balloon angioplasty, we determined genotypes for 94 SNPs representing 62 candidate genes, in a prospectively assembled cohort of 342 cases and 437 controls. Using a customized coupled-logistic regression procedure accounting for both additive and interactive effects, we identified seven SNPs in seven genes that, together, showed a statistically significant association with restenosis incidence (P <0.0001), accounting for 11.6% of overall variance observed. Among them are candidate genes for cardiovascular pathophysiology (apolipoprotein species and NOS), inflammatory response (TNF receptor and CD14), and cell-cycle control (p53 and p53-associated protein). Our results emphasize the need to account for complex multi-gene influences and interactions when assessing the molecular pathology of multifactorial medical entities. PMID- 12082595 TI - The epidemiology of hypertension and its associated risk factors in the city of Debrecen, Hungary. AB - The purpose of this work was to estimate the prevalence of hypertension and cardiovascular risk factors and its association with sociodemographic, behavioural and lifestyle characteristics among the adult population of the city of Debrecen, Hungary.A cross-sectional study was conducted in 1996. Amongst 21 800 inhabitants aged 30-65 y risk screening for cardiovascular disease (CVD) was estimated by a questionnaire that included sociodemographic, lifestyle characteristics, family history of CVD and results of self-reported data of body weight, height and blood pressure. Of the total of 19 961 surveyed sample, 37.02% were considered to be hypertensive, 53.73% were overweight, 32.18% were current smokers and 58.85% were physically inactive. Hypertensives were older than normotensives (50.81+/-9.01 vs 44.78+/-8.97 y, P<0.001). The prevalence of various risk factors amongst hypertensives as compared to normotensives were overweight (68.49 vs 45.06%, P<0.0001), current smoking (28.38 vs 34.41%, P<0.0001), physical inactivity (64.78 vs 55.36%, P<0.001), and high alcohol consumption (1.91 vs 1.06%, P<0.01). Of the hypertensives 37.11% were on drug therapy. Of those on therapy, only 17.03% had normal blood pressure. Being overweight and having low physical activity was positively associated with hypertension (OR=2.25, CI=2.11-2.40) and (OR=1.26, CI=1.15-1.38). Manual work, a family history of CVD, low education, and the male gender were also associated with hypertension and more CVD risk factors. These findings illustrate a very high prevalence of hypertension and CVD risk factors in Debrecen, indicating the importance of the need for more effective prevention programmes and control of hypertension in Hungary. PMID- 12082596 TI - The role of demographic surveillance systems (DSS) in assessing the health of communities: an example from rural Ethiopia. AB - Longitudinal demographic surveillance systems (DSSs) in selected populations can provide important information in situations where routine health information is incomplete or absent, particularly in developing countries. The Butajira Rural Health Project is one such example, initiated in rural Ethiopia in 1987. DSSs rely on regular community-based surveillance as a means of vital event registration, among a sufficient population base to draw meaningful conclusions about rates and trends in relatively rare events such as maternal death. Enquiries into specific health problems can also then use this framework to quantify particular issues or evaluate interventions. Demographic characteristics and trends for a rural Ethiopian population over a 10-y period are presented as an illustration of the DSS approach, based on 336 000 person-years observed. Overall life expectancy at birth was 50 y. Demographic parameters generally showed modest trends towards improvement over the 10-y period. The DSS approach is useful in characterising populations at the community level over a period of time, providing important information for health planning and intervention. Methodological issues underlying this approach need further exploration and development. PMID- 12082597 TI - Narrowing the gap between academic professional wisdom and community lay knowledge: perceptions from partnerships. AB - Community involvement in health through community partnerships (CPs) has been widely advocated. Putting CPs into practice is complex and represents a challenge for all the stakeholders involved in the change process. Employing data from five CPs aiming to bring together communities, academics and health service providers in South Africa, this paper aims to examine and compare the views of the health care professionals with those of the community members with respect to each other's skills and abilities. Five domains of expertise in partnership working are examined: educational competencies; partnership fostering skills; community involvement expertise; change agents proficiencies; and strategic and management capacities. The findings suggest that the community recognizes the expertise and abilities brought by the professional staff to the CPs. Community members have a positive view of the capabilities of the professionals, in particular their abilities as resource persons in the areas of budget management, policy formulation and the introduction and management of change. The professionals, on the other hand, are cautious regarding the level of skill and capability in communities. The limited appreciation of community skills by the professionals covered all the five domains of expertise examined. The findings suggest that if joint working is to survive, the professionals will need to increase their valuation of the indigenous proficiencies inherent in their community partners. We conclude that programme models need to consciously incorporate in their design and implementation, capacity building, skills transfer and empowerment strategies. PMID- 12082598 TI - Survival in a cohort of social services placements in nursing and residential homes: factors associated with life expectancy and mortality. AB - The aim of this study was to examine the life expectancy of elderly people in nursing and residential care over a 20-month period and its relationship to specific risk factors. Using a retrospective cohort design, data obtained on 1888 residents placed between 1 July 1997 and 30 April 1999 in residential, nursing and dual registered homes within Nottingham Health Authority boundaries were examined. Additional data on physical and mental disability at placement were available for 514 residents. Main outcome measures comprised survival rate overall, and in relation to gender, age, home type (nursing, residential or dual), source of placement (hospital or community) and various disability factors.One-year survival rates were: overall, 66%; nursing homes, 59%; dual homes, 58%; and residential homes 76%. Median survival in nursing homes was 541 days, but was not reached in residential homes. Male gender, admission to nursing or dual registered homes, placement from hospital, decreased mobility and increased age were associated with decreased life expectancy. Although no association was found between length of survival and level of cognitive function, lack of cognitive impairment was associated with lower survival. In conclusion, mortality is high in nursing, dual and residential homes where life expectancy has been shown to be associated with gender, home type, origin of placement and mobility. Rates of survival are related to higher comorbidity and disability. Important data for planning and assessing care needs can be yielded through the analysis of mortality data. PMID- 12082599 TI - Differences between Japanese pre-school and school-age pedestrian mortality and morbidity trends. AB - The risk of pedestrian injury during childhood in industrialised countries is decreasing in a trend that is related to exposure to traffic. Examining the differences between age groups and their behaviour provides an insight into exposure to risks. Using data for the period 1968-1998, we compared the changes in pedestrian mortality and morbidity in pre-school children (aged 0-6) with those of school-age children (aged 7-12). Pedestrian mortality and morbidity decreased in children of all ages in the early 1970s. However, after the mid 1970s, both mortality and morbidity continued to decrease only in pre-schoolers. In school-age children, mortality continued to decrease but morbidity did not change. These age-related differences in mortality and morbidity indicate that over the past 30 y the environment for child pedestrians did not become safer. Instead, some of the decrease in children's injuries was probably achieved at the expense of their outdoor activities. Efforts to protect children from vehicular traffic should focus on changing the outdoor environment rather than on restricting children's activities. PMID- 12082601 TI - Case mix presenting to paediatricians in a UK district (1998). AB - This observational study analysed paediatric outpatient referrals to a district service in the UK in 1998 and compared results with studies conducted 10 y previously. Data was collected prospectively from the written correspondence on all new outpatient referrals offered appointments by paediatricians at Northampton General Hospital over a 3-month period. Two hundred and ninety-seven (66%) referrals were to general paediatricians, 108 (24%) to 'patch' community paediatricians and 47 (10%) to community paediatrician consultant clinics. The referral rate to general outpatients was 20/1000 per annum, similar to referral rates in 1988. Urogenital and cardiac problems have overtaken asthma as the most commonly referred conditions. Referrals to consultant community paediatricians were predominantly development and behaviour difficulties and were more complex than those referred to other groups. 'Patch' community paediatricians received 87% of their referrals from practitioners other than doctors and played a significant role in co-ordinating care in the community. They referred less than 3% for consultant assessment. The referral rate to general paediatric outpatients has remained stable over the last decade although the case mix has changed. For the first time, we provide population-based case mix and referral rates to community paediatricians. We highlight the important role of non-consultant career grade paediatricians in supporting clinical services in the community. Open access to some investigations for general practitioners (GPs) could be explored as a way of managing demand for general paediatric outpatient appointments. PMID- 12082600 TI - Bullying in Jerusalem schools: victims and perpetrators. AB - The objectives of this study were to determine the prevalence of bullying among adolescents studying in Jerusalem schools and to identify the characteristics of its victims and perpetrators. Pupils in the 8th and 10th grade (ages 14-16 y) in 11 schools in Jerusalem (n=1182) anonymously completed the World Health Organization questionnaire from the 'Health Behavior in School Children' study. Bullying others or being bullied at least once in the last term was reported. The independent variables studied were socio-demographic characteristics, and personal and school-related factors. Bullying was reported by 57.1% of boys and 27.0% of girls, and being bullied by 50.3% of boys and 39.5% of girls. The factors associated with bullying others were lack of support from teachers for both genders and poor mental health among boys. Variables related to social exclusion among girls and social isolation among boys were associated with being bullied. Health professionals should be active in all levels of prevention to deal with this problem that affects the health and well-being of so many children and adolescents. PMID- 12082602 TI - Personal View: health protection: the final straight? PMID- 12082605 TI - Prevalence of health and social care bed occupancy by marital status. PMID- 12082606 TI - Nbs1 promotes ATM dependent phosphorylation events including those required for G1/S arrest. AB - Cell lines from Nijmegen Breakage Syndrome (NBS) and ataxia telangiectasia (A-T) patients show defective S phase checkpoint arrest. In contrast, only A-T but not NBS cells are significantly defective in radiation-induced G1/S arrest. Phosphorylation of some ATM substrates has been shown to occur in NBS cells. It has, therefore, been concluded that Nbs1 checkpoint function is S phase specific. Here, we have compared NBS with A-T cell lines (AT-5762ins137) that express a low level of normal ATM protein to evaluate the impact of residual Nbs1 function in NBS cells. The radiation-induced cell cycle response of these NBS and 'leaky' A-T cells is almost identical; normal G2/M arrest after 2 Gy, intermediate G1/S arrest depending on the dose and an A-T-like S phase checkpoint defect. Thus, the checkpoint assays differ in their sensitivity to low ATM activity. Radiation induced phosphorylation of the ATM-dependent substrates Chk2, RPAp34 and p53 Ser15 are similarly impaired in AT-5762ins137 and NBS cells in a dose dependent manner. In contrast, NBS cells show normal ability to activate ATM kinase following irradiation in vitro and in vivo. We propose that Nbs1 facilitates ATM dependent phosphorylation of multiple downstream substrates, including those required for G1/S arrest. PMID- 12082607 TI - Multiple stages of malignant transformation of human endothelial cells modelled by co-expression of telomerase reverse transcriptase, SV40 T antigen and oncogenic N-ras. AB - We have modelled multiple stages of malignant transformation of human endothelial cells (ECs) by overexpressing the catalytic subunit of human telomerase (hTERT), together with SV40 T antigen (SV40T) and oncogenic N-ras. Transfection with hTERT alone, led to the immortalization of two out of three cultures of bone marrow derived ECs (BMECs). One hTERT transduced BMEC culture underwent a long proliferative lag before resuming proliferation. BMECs transfected with hTERT alone were functionally and phenotypically normal. BMECs transfected with SV40T (BMSVTs) had an extended lifespan, but eventually succumbed to crisis. BMSVTs exhibited a partially transformed phenotype, demonstrating growth factor independence, altered antigen expression and forming tiny, infrequent colonies in vitro. Transduction of BMSVTs with hTERT resulted in immortalization of 4 out of 4 cultures. BMSVTs immortalized with hTERT formed large colonies in vitro and small transient tumours in vivo. BMECs co-expressing SV40T, hTERT and N-ras exhibited an overtly transformed phenotype; forming very large colonies with an altered morphology and generating rapidly growing tumours in vivo. These investigations demonstrate transformation of human ECs to an overtly malignant phenotype. This model will be useful for understanding mechanisms underlying vascular and angiogenic neoplasias, as well as for testing drugs designed to curtail aberrant EC growth. PMID- 12082608 TI - Anoxia induces macrophage inhibitory cytokine-1 (MIC-1) in glioblastoma cells independently of p53 and HIF-1. AB - Human astrocytic brain tumors select for mutations in the p53 tumor suppressor gene early in malignant progression. p53 is activated upon various kinds of cellular stress leading to apoptosis or cell cycle arrest, but is also implicated in complex biological processes such as inhibition of angiogenesis and metastasis. In an effort to shed light on consequences mediated by p53 inactivation in gliomas, we established the Tet-On system for p53 in the LN-Z308 glioblastoma cell line. The macrophage inhibitory cytokine-1 (MIC-1) gene was identified as a most prominent p53 target gene upon gene expression profiling. Oxygen deprivation, an important cellular stress, revealed MIC-1 as an anoxia responsive gene in glioblastoma cell lines. MIC-1 up-regulation by anoxia is mediated through an alternative, p53 and hypoxia inducible factor 1 (HIF-1) independent pathway. Furthermore, ectopic expression of MIC-1 in LN-Z308 cell line completely abolished its inherent tumorigenicity in nude mice, while proliferation in vitro was not affected. In the present experimental model MIC-1 may exert its anti-tumorigenic properties via a paracrine mechanism mediated by host cells in vivo. Taken together, these data suggest that MIC-1 is an important downstream mediator of p53 function, while acting itself as an intercessor of cellular stress signaling and exerting anti-tumorigenic activities. PMID- 12082609 TI - p14(ARF) regulates E2F activity. AB - The ARF protein product of the ink4a/arf locus is induced by a variety of oncogenic signals. ARF facilitates growth arrest through the p53 pathway by hindering the down-regulation of p53 activity mediated by MDM2, through the formation of a protein complex with MDM2. Here we have explored the possibility that human p14(ARF) activity is integrated with growth regulating pathways other than p53, and report our results that p14(ARF) can control the activity of the E2F transcription factor. p14(ARF) regulates E2F activity in different cell types, including p53(-/-)/mdm(-/-) MEFs, thus excluding that the effects of p14(ARF) are indirectly caused through MDM2 modulation. p14(ARF) down-regulates E2F-dependent transcription, and in cells undergoing E2F-dependent apoptosis prompts cell cycle arrest. p14(ARF) possesses multiple binding domains for E2F-1, one of which resides within the N-terminal region and coincides with the regulation of E2F activity. A mutational analysis of p14(ARF) indicates that the E2F-1 and MDM2 binding domains can be distinguished. These results highlight the potential interplay between p14(ARF) and E2F, and establish p14(ARF) as a pleiotrophic regulator of cell growth that acts by targetting at least two key pathways in the control of proliferation, namely E2F and p53. PMID- 12082610 TI - Patterns of gene promoter methylation in squamous cell cancer of the head and neck. AB - Promoter methylation is an important pathway in transcriptional silencing of known and candidate tumor suppressor genes in Head and Neck Squamous Cell Carcinoma (HNSCC). In order to study the association of tumor suppressor gene promoter methylation in HNSCC with patient clinical characteristics, especially alcohol consumption and tobacco smoking, we examined promoter methylation of the p16(INK4a), DAP-kinase, E-Cadherin, and RASSF1A genes using methylation-specific PCR (MSP) in 80 patients. The prevalence of p16(INK4a), DAP-kinase, E-Cadherin, and RASSF1A promoter methylation was 26/80 (32.5%), 19/80 (23.8%), 29/80 (36.3%), 6/80 (7.5%) respectively. In 48 cases (60%), at least one of these promoters was methylated. There was a significant association of methylation of any of these genes and ever smoking (P=0.006). p16(INK4a) gene promoter methylation was associated with a younger age of smoking initiation (P<0.03); E-Cadherin promoter methylation was associated with an increased number of pack years smoked (P<0.03). We also found an association of methylation of any gene and T status (OR=2.7, P<0.05). Tumors with p16(INK4a) methylation were significantly less likely to show lymph node metastasis (P<0.001). DAP-kinase promoter methylation was significantly associated with lymph node metastasis and this relationship was dependent upon p16(INK4a) promoter methylation status. Our results suggest that, in HNSCC, promoter methylation of these four genes accumulates with increasing tumor size. This may reflect distinct pathways of somatic inactivation leading to cancer; additional larger studies are needed to further investigate this possibility. Tobacco smoking may play an important role in both the occurrence of promoter methylation as well as delineating the precise pathway that eventually results in a tumorigenic phenotype. PMID- 12082611 TI - Multiple tristetraprolin sequence domains required to induce apoptosis and modulate responses to TNFalpha through distinct pathways. AB - Expression of the immediate early protein tristetraprolin (TTP) is induced by numerous stimuli, including tumor necrosis factor-alpha (TNFalpha). Evidence indicates that TTP limits production of TNFalpha and other cytokines by directly binding and destabilizing their mRNAs. This effect seems to require only the conserved TTP zinc finger region, and is characteristic of the related proteins TIS11b and TIS11d. TTP, TIS11b, and TIS11d each also induce apoptosis through the mitochondrial pathway analogously to certain oncogenes, suggesting that they influence growth or survival signals. Among TTP/TIS11 proteins, TTP alone also promotes apoptosis synergistically with TNFalpha. Here we show that other regions of TTP along with the zinc fingers are required for TTP to induce apoptosis. We also demonstrate that TTP acts through an additional pathway to sensitize cells to the pro-apoptotic stimulus of TNFalpha. This modulation of TNFalpha responses specifically requires the TTP N-terminal region, which is not conserved in TIS11b or TIS11d. We conclude that the physiological functions of TTP depend upon multiple regions of the TTP protein, that TTP has diverged functionally from TIS11b and TIS11d, and that modulation of TNFalpha responses may be a unique and important aspect of TTP function. PMID- 12082612 TI - Nup98-HoxA9 immortalizes myeloid progenitors, enforces expression of Hoxa9, Hoxa7 and Meis1, and alters cytokine-specific responses in a manner similar to that induced by retroviral co-expression of Hoxa9 and Meis1. AB - The association between acute myeloid leukaemia (AML) and the aberrant expression of Hoxa9 is evidenced by (1) proviral activation of Hoxa9 and Meis1 in BXH-2 murine AML, (2) formation of the chimeric Nup98-HoxA9 transactivator protein as a consequence of the t(7;11) translocation in human AML, and (3) the strong expression of HoxA9 and Meis1 in human AML. In mouse models, enforced retroviral expression of Hoxa9 alone in marrow is not sufficient to cause rapid AML, while co-expression of Meis1 and Hoxa9 induces rapid AML. In contrast, retroviral expression of Nup98-HoxA9 is sufficient to cause rapid AML in the absence of enforced Meis1 expression. Previously, we demonstrated that Hoxa9 could block the differentiation of murine marrow progenitors cultured in granulocyte-macrophage colony-simulating factor (GM-CSF). These progenitors lacked Meis1 expression, could not proliferate in stem cell factor (SCF), but could differentiate into neutrophils when switched into granulocyte colony-simulating factor (G-CSF). Ectopic expression of Meis1 in these Hoxa9 cells suppressed their G-CSF-induced differentiation, permitted proliferation in SCF, and therein offered a potential explanation of cooperative function. Because Meis1 binds N-terminal Hoxa9 sequences that are replaced by Nup98, we hypothesized that Nup98-HoxA9 might consolidate the biochemical functions of both Hoxa9 and Meis1 on target gene promoters and might evoke their same lymphokine-responsive profile in immortalized progenitors. Here we report that Nup98-HoxA9, indeed mimicks Hoxa9 plus Meis1 coexpression - it immortalizes myeloid progenitors, prevents differentiation in response to GM-CSF, IL-3, G-CSF, and permits proliferation in SCF. Unexpectedly, however, Nup98-Hoxa9 also enforced strong transcription of the cellular Hoxa9, Hoxa7 and Meis1 genes at levels similar to those found in mouse AML's generated by proviral activation of Hoxa9 and Meis1. Using Hoxa9(-/-) marrow, we demonstrate that expression of Hoxa9 is not required for myeloid immortalization by Nup98-HoxA9. Rapid leukaemogenesis by Nup98-HoxA9 may therefore result from both the intrinsic functions of Nup98-HoxA9, as well as of those of coexpressed HOX and MEIS1 genes. PMID- 12082613 TI - TNF-Selectokine: a novel prodrug generated for tumor targeting and site-specific activation of tumor necrosis factor. AB - We describe a TNF fusion protein designated TNF-Selectokine, which is a homo trimeric molecule comprised of a single chain antibody (scFv) targeting module, a trimerization domain and TNF. TNF-Selectokine exerts high bioactivity towards the targeted and adjacent, antigen negative cells. Membrane targeting dependent immobilization of the TNF-Selectokine induced cell death in TNFR1 and TNFR2 dependent manner, thus cell bound TNF-Selectokine mimicks membrane TNF. To restrict TNF activity to the tumor, a prototype of a TNF-Selectokine prodrug was constructed by insertion of a TNFR1 fragment, separated from TNF by a protease sensitive linker. The prodrug exerts minimal TNF activity, but can be activated in vitro several thousand-fold by proteolytic digest, showing the principal feasibility of this approach. Choice of cleavage site(s) recognized by protease(s) typically associated with a given carcinoma should allow high dose systemic application of the respective TNF prodrug that unveils its specific bioactivity only in targeted tissues. PMID- 12082614 TI - Calgranulins S100A8 and S100A9 are negatively regulated by glucocorticoids in a c Fos-dependent manner and overexpressed throughout skin carcinogenesis. AB - The two calgranulins S100A8 and S100A9 were found to be differentially expressed at sites of acute and chronic inflammation. Here we have employed the phorbol ester-induced multistage skin carcinogenesis protocol in mice to determine the expression of both genes in inflamed skin and in skin tumors. We show that expression is coordinately induced by the phorbol ester TPA in epithelial cells as well as infiltrating leukocytes. By comparing S100A8 and S100A9 mRNA levels in wild type and c-Fos deficient mice (c-fos(-/-)) we found that expression is negatively regulated by c-Fos/AP-1. Glucocorticoids, which exhibit potent anti inflammatory and anti-tumor promoting activities repressed TPA-mediated S100A8 and S100A9 induction in wild type, but not in c-fos(-/-) mice, thus identifying both genes as the first examples of AP-1 target genes whose repression of TPA induced transcription by glucocorticoids depends on c-Fos. Finally, we show that enhanced expression is not restricted to the initial TPA-induced inflammatory response but is observed at all stages of skin carcinogenesis. These data identify S100A8 and S100A9 as novel, tumor-associated genes and may point to an as yet unrecognized function of both genes in the development of epithelial skin tumors. PMID- 12082615 TI - Sequential extension of proliferative lifespan in human fibroblasts induced by over-expression of CDK4 or 6 and loss of p53 function. AB - Replicative senescence is thought to be a significant barrier to human tumorigenesis, which in human fibroblasts, and many other cell types, can be overcome experimentally by combined loss of function of p53 and Rb 'pathways'. To avoid the confounding pleiotropic effects of HPVE7 frequently used in such studies, here we have employed retroviral vectors over-expressing CDK4 or CDK6 as a more representative model of naturally-occurring mutations targeting the Rb pathway. We show that these can extend fibroblast lifespan by approximately 10 population doublings, ending in a viable senescence-like state which contrasts with the apoptotic end-stage seen with E7. Compared with 'normal' senescence, this growth arrest was, in most cases, not accompanied by any further increase in p21(Waf1) levels but with up to a 19-fold increase in p16(Ink4a). Surprisingly however, this could not explain arrest, since expression of mutant CDK4 and/or CDK6, incapable of binding p16(Ink4a), did not confer any greater lifespan extension than the wild-type CDKs. Subsequent abrogation of p53 function by a second vector, encoding HPVE6, downregulated p21(Waf1) and conferred a second lifespan extension, ending in a crisis-like state, consistent with full escape from senescence. These data: (i) point to a back-up 'senescence' mechanism distinct from induction of p21(Waf1) or p16(Ink4a); and (ii) provide an in vitro model of clonal evolution through successive dysfunction of Rb and p53 pathways in a relevant human cell context. PMID- 12082617 TI - Common occurrence of multiple K-RAS mutations in pancreatic cancers with associated precursor lesions and in biliary cancers. AB - Recent studies on small series of pancreatic cancer (PC) with foci of pancreatic intraepithelial neoplasia (PanIN), a putative precursor lesion, have shown that multiple K-RAS mutations may coexist in the same neoplastic pancreas. To see whether mutant-K-RAS polyclonality is a common and specific feature of pancreatic carcinogenesis, we investigated a unselected series of periampullary cancers (41 pancreatic, 13 biliary and two ampullary adenocarcinomas). After hemi-nested polymerase chain reaction (PCR), mutations identified with single strand conformation polymorphism (SSCP) were confirmed by allele-specific PCR and sequencing. K-RAS codon 12 was mutated in 34 (83%) pancreatic cancers and in 11 (85%) biliary cancers. Multiple distinct K-RAS mutations were found in 16 PC (39% of all cases, 47% of those with mutated K-RAS) and in eight biliary cancers (62 and 72%, respectively). In PC, multiple K-RAS mutations were more frequent (P<0.001) in cancers with (nine of 12, 75%) than in those without detectable PanIN (seven of 29, 24%). Individual precursor lesions of the same neoplastic pancreas were found to harbor distinct mutations. Results show that multiple K RAS mutations are frequent both in PC with associated PanIN and in biliary cancers, and indicate that clonally distinct precursor lesions of PC might variably contribute to tumor development. PMID- 12082616 TI - Expression and transactivating functions of the bZIP transcription factor GADD153 in mammary epithelial cells. AB - Heregulin-beta1 (HRG), a combinatorial ligand for human epidermal growth factor receptor 3 (HER3) and HER4, is a regulatory polypeptide having distinct biological effects, such as growth stimulation, differentiation, invasiveness, and migration in mammary epithelial cells. The mechanism underlying the diverse functions of HRG is not well established but is believed to depend on induced changes in the expression of specific cellular gene products, their modification, or both. Here, we identified the basic leucine zipper transcription factor, the growth-arrest and DNA-damage 153 (GADD153)/CCAAT-enhancer binding protein (C/EBP) homologous protein (CHOP) as one of the HRG-inducible genes. We demonstrated that HRG stimulation of mammary epithelial cells induces the expression of GADD153 mRNA and protein and transcription of GADD153 promoter. The transcriptional activity of the GADD153 promoter as well as transcription from the C/EBP activating transcription factor (ATF) composite motif in the GADD153 promoter was also stimulated by HRG-inducible ATF-4 and activated HER2 but not wild-type HER2. GADD153 expression was upregulated by the lactogenic hormones insulin and progesterone and associated with differentiation of normal mammary epithelial cells. Consistent with its role as transcriptional modifier, GADD153 stimulated transcription of beta-casein promoter activity in a STAT5a-sensitive manner in mammary epithelial cells. Analysis of mouse mammary gland development revealed that GADD153 expression was predominantly restricted in the early lactating stages. Because cyclic AMP responsive element and ATF binding sites are present in a variety of growth-regulating cellular genes, these findings suggest that stimulation of GADD153 expression and its transactivating functions may constitute an important mechanism of regulation of putative genes having diverse functions during cell growth and differentiation. PMID- 12082618 TI - Expression of thyroid hormone receptor/erbA genes is altered in human breast cancer. AB - The relation between thyroid status and diseases and cancer is unclear. No detailed analysis of thyroid hormone receptor (TR) expression in human breast cancer has been reported. We have analysed the expression and mutational status of the TRalpha1, encoded by the c-erbA proto-oncogene, TRbeta1 and TRbeta2 isoforms in 70 sporadic breast cancers. Alterations in the RNA level of TRbeta1, TRalpha1, or both were found in a number of patients. No expression of TRbeta2 RNA was detected. Western blotting analysis confirmed the differences in expression at the protein level in those cases where sufficient tumor sample was available. Additionally, tumor-specific truncated TRbeta1 RNA was found in six patients. Strikingly, three transcripts shared the same breakpoint. Only one tumor carried the corresponding deletion at the genomic DNA level, suggesting that the remaining abnormal TRbeta1 transcripts are aberrant splicing products. Though no significant correlation was found between TRbeta1 alteration and any clinical parameter, it showed a tendency to associate with early age of onset (<50 years). Our results reveal specific alterations in the expression of TRbeta and TRalpha genes in a subset of breast cancer patients, suggesting that deregulation of thyroid hormone target genes may be involved in the generation of this neoplasia. PMID- 12082619 TI - Increased choline kinase activity in human breast carcinomas: clinical evidence for a potential novel antitumor strategy. AB - Choline kinase (ChoK) and its product, phosphocholine (PCho), have been implicated in human carcinogenesis. Inhibition of this enzyme has been shown to be an efficient antitumor strategy in vivo. The aim of this study was to assess the relationship between the regulation of ChoK and clinical features in patients with breast cancer. To that end, normal and tumoral tissues from 53 patients with breast carcinomas were analysed for ChoK activity and expression, and compared to some clinical parameters. We report a relevant increase in ChoK activity in 38.5% of the tumoral tissues analysed when compared to the normal levels in healthy tissues. Furthermore, some clinical features were found significant versus ChoK status. First, an association of choline enzymatic activity with histological tumor grade was observed (P<0.001). In addition, increased ChoK activity was also associated with ER-negative breast carcinomas (P=0.037). A significant association between ChoK overexpression and both high histologic tumor grade (P=0.017) and ER-negative tumors (P=0.003) was found. Finally, ChoK overexpression was found in 17% of the samples and all corresponded with those that display the highest increase in ChoK activity (P<0.001). Here we provide evidence that ChoK may be related to the development of human breast cancer, suggesting that this finding may constitute the basis for the development of a novel antitumoral strategy for these patients. PMID- 12082620 TI - Stimulation of the mitogen-activated protein kinase pathway antagonizes TRAIL induced apoptosis downstream of BID cleavage in human breast cancer MCF-7 cells. AB - We studied the role of the mitogen-activated protein kinase (MAPK) pathway in the regulation of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) mediated apoptosis in breast tumor MCF-7 cells. We found that addition of a protein kinase C (PKC) activator to MCF-7 cultures prevented TRAIL-induced apoptosis, by inhibiting a step downstream of both caspase-8 activation and BID cleavage. TRAIL-induced translocation of Bax from cytosol to mitochondria, release of cytochrome c from mitochondria and activation of caspase-9 were all inhibited by PKC activation. PKC-mediated prevention of mitochondrial apoptotic events and apoptosis was found to be dependent on the MAPK pathway. Since TRAIL is a ligand of potential use in antineoplastic clinical trials, our findings may provide relevant information in cancer therapy. PMID- 12082621 TI - Clusterin (SGP-2) transient overexpression decreases proliferation rate of SV40 immortalized human prostate epithelial cells by slowing down cell cycle progression. AB - Clusterin is a highly conserved, widely distributed glycoprotein whose biological significance is still debated. Involved in many biological processes and disease states, clusterin is induced by cell injury and tissue regression, but is repressed during cell proliferation. We have previously reported that clusterin mRNA induction is associated with epithelial cell atrophy in the rat prostate and both clusterin transcript and protein accumulated in quiescent normal human skin fibroblasts. Here we show that transient clusterin overexpression, in SV40 immortalized human prostate epithelial cells (PNT2), resulted in increased accumulation of cells in the G(0)/G(1) phases of the cell cycle, accompanied by slowdown of cell cycle progression and decrease of DNA synthesis. The activities of ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (AdoMetDC), and the level of histone H3 mRNA (markers of cell proliferation) concomitantly decreased, while Gas1 mRNA (a marker of cell quiescence) accumulated. Thus it appears that clusterin, by opposing the effect of SV40 on the proliferation rate of PNT2 cells, acts as an anti-oncogene in the prostate, suggesting a role for this gene in controlling proliferation of normal and transformed prostate epithelial cells. PMID- 12082622 TI - Loss of Stat5a delays mammary cancer progression in a mouse model. AB - A genetic study was conducted to determine if activated Stat5a contributes to mammary carcinogenesis and to evaluate the mechanism. Similar to human breast cancers, a proportion of mammary adenocarcinomas in the WAP-TAg transgenic mouse model demonstrate constitutive Stat5a/b and Stat3 activation. Stat5a activation is linked to mammary epithelial cell survival and differentiation, and proliferation in hematopoetic cell lineages. Breeding WAP-TAg mice to mice carrying germ-line deletions of the Stat5a gene generated mice with reduced levels of Stat5a. Hemizygous loss of the Stat5a allele significantly reduced levels of Stat5a expression without altering mammary gland development or transgene expression levels. In comparison to mice carrying two wild-type Stat5a alleles, hemizygous loss of the Stat5a allele reduced the number of mice with palpable tumors at 7 months of age (67% from 100%, P<0.05), resulted in smaller tumors at 7 months of age (3.8 cm3 from 7.6 cm3, P=0.003), delayed first tumor appearance (208 days from 188 days, P=0.01), and increased the apoptotic index in the adenocarcinomas (4.3+/-0.3 from 1.2+/-0.2, P=0.016). Neither cell proliferation nor differentiation in the cancers was altered. Decreasing Stat5a activation levels could be a therapeutic approach for reducing survival of breast cancer cells. PMID- 12082623 TI - Progressive aberrant methylation of the RASSF1A gene in simian virus 40 infected human mesothelial cells. AB - Mesotheliomas are tumors arising from mesothelial cells and are associated with asbestos exposure and approximately 50% contain simian virus 40 (SV40) DNA sequences. SV40 infection of human mesothelial cells (HM) causes early cellular immortalization and late transformation. Aberrant methylation is a major mech anism for loss of function of tumor suppressor genes (TSGs). We recently reported that of seven genes frequently methylated in epithelial tumors, only RASSF1A gene was frequently methylated in mesotheliomas, and its methylation was correlated with loss of RASSF1A expression and the presence of SV40. We studied whether SV40 infection of normal HM induces aberrant methylation of the genes previously studied in mesotheliomas. Of six infected foci examined at early passages (passages 8-30) there was no methylation of the seven genes examined. Of two foci examined at late passages (passages 51-86) after the appearance of morphological changes suggestive of transformation, methylation and loss of expression of RASSF1A was detected. Sequencing of the CpG dense region around the transcription start site and semi-quantitative real-time methylation specific PCR (MSP) assay for RASSF1A methylation demonstrated progressive methylation during late passages. Exposure to the demethylating agent 5-aza-2'-deoxycytidine restored RASSF1A expression, while exposure to the histone deacetylation inhibitor trichostatin A had no effect. These data, together with our previous findings, support a causal relationship between SV40 infection, progressive RASSF1A methylation and its silencing, and the pathogenesis of mesothelioma. PMID- 12082624 TI - Aberrant promoter methylation and silencing of the RASSF1A gene in pediatric tumors and cell lines. AB - Aberrant promoter methylation of tumor suppressor genes has not been fully investigated in pediatric tumors. Therefore, we examined the methylation status of nine genes (p16(INK4A), MGMT, GSTP1, RASSF1A, APC, DAPK, RARbeta, CDH1 and CDH13) in 175 primary pediatric tumors and 23 tumor cell lines using methylation specific PCR. We studied the major forms of pediatric tumors--Wilms' tumor, neuroblastoma, hepatoblastoma, medulloblastoma, rhabdomyosarcoma, osteosarcoma, Ewing's sarcoma, retinoblastoma and acute leukemia. The most frequently methylated gene in both primary tumors and cell lines was RASSF1A (40, 86%, respectively). However, the rates of RASSF1A methylation in individual tumor types varied from 0 to 88%. RASSF1A methylation was tumor specific and was absent in adjacent non-malignant tissues. Methylation of the other genes was relatively rare in tumors and non-malignant tissues (less than 5%). Neuroblastoma patients with methylation of RASSF1A were significantly older than patients without methylation (P=0.008). There was no relationship between methylation status and other clinico-pathologic parameters. We treated six cell lines lacking RASSF1A mRNA with 5-aza-2'deoxycytidine to examine the relationship between methylation and transcriptional silencing. In five of six cell lines, restoration of RASSF1A mRNA was confirmed by RT-PCR. Our findings indicate that aberrant promoter methylation of RASSF1A may contribute to the pathogenesis of many different forms of pediatric tumors. PMID- 12082625 TI - Aurora-B associated protein phosphatases as negative regulators of kinase activation. AB - The human serine/threonine kinase Aurora-B is structurally related to the protein kinase Ipl1p from S cerevisiae and aurora from Drosophila melanogaster, which are key regulators of mitosis. The present study shows that human Aurora-B is activated by okadaic acid and forms complexes with the protein serine/threonine phosphatase type 1 (PP1) or PP2A, but not with PP5. These data identified Aurora B associated protein phosphatases as negative regulators of kinase activation. We then used a series of substrates based on a histone H3 phosphorylation site (residues 5-15) to determine the substrate specificity of human Aurora-B. We found that this enzyme is an arginine-directed kinase that can phosphorylate histone H3 at serines 10 and 28 in vitro, suggesting that human Aurora-B is a mitotic histone H3 kinase. PMID- 12082626 TI - Chromatin remodeling factor encoded by ini1 induces G1 arrest and apoptosis in ini1-deficient cells. AB - Ini1/hsnf5 gene encodes INI1 protein, a chromatin remodeling factor associated with the SWI/SNF complex. In yeast, this complex modifies chromatin condensation to coactivate various transcriptional factors. However, in human, little is known about the SWI/SNF complex and INI1. To elucidate cellular functions of ini1, we constructed a recombinant adenovirus (AdexHA-INI1) capable of overexpressing INI1 in ini1-deficient cells. AdexHA-INI1 produced intranuclear INI1 in three ini1 deficient cell lines, changed their morphology, and decreased the proportion of viable cells. Flow cytometry and a BrdU incorporation assay showed that after the infection, growth of these cells was partially arrested at G1. In two of the three ini1-deficient cell lines, apoptosis was found to occur after the infection, as detected by the presence of cleaved poly (ADP-ribose) polymerase. To determine functional domains of INI1, we constructed plasmids expressing INI1 and its deletion mutants, which were used for a colony formation assay. Repeats 1 and 2 of INI1 were found to be required to suppress the growth of the three ini1 deficient cell lines. The results support the hypothesis that ini1 is a tumor suppressor gene and suggest a novel link between human SWI/SNF chromatin remodeling complex and apoptosis. PMID- 12082627 TI - Evidence that the human death receptor 4 is regulated by activator protein 1. AB - Death receptor 4 (DR4; also called TRAIL-R1), a member of the tumor necrosis factor receptor superfamily, is a cell surface receptor that triggers the apoptotic machinery upon binding to its ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Although several chemotherapeutic agents were reported to induce DR4 expression, the mechanism of this effect remains largely unknown. To begin to understand its regulation, we cloned a 1.8 Kb 5'-flanking region of the human DR4 gene and identified several putative binding sites for transcription factors including activator protein 1 (AP-1). Among the three putative AP-1 binding sites, the site located at -350/-344 is functionally active as evidenced by a combination of electrophoretic mobility shift and luciferase reporter assays. The AP-1 activator phorbol 12-myristate 13-acetate (TPA) enhanced the binding of this DR4 AP-1 binding site to protein(s) in a nuclear extract from TPA-treated cells, increased luciferase activity of a reporter construct containing this site and induced DR4 expression at the transcription level. These results indicate that AP-1 regulates DR4 expression via the AP-1 binding site located at -350/-344. AP-1 has been implicated in many critical cellular processes including apoptosis, and is a major target of the c-Jun NH(3) terminal kinase signaling pathway that is activated by many anticancer drugs. Therefore, our findings may increase the understanding of the mechanisms underlying AP-1-mediated apoptosis as well as drug-induced apoptosis. PMID- 12082628 TI - TERT regulates cell survival independent of telomerase enzymatic activity. AB - Human telomerase reverse transcriptase (hTERT), the catalytic subunit of telomerase, plays a pivotal role in the maintenance of telomeres and cell proliferation. Here we report that down-regulation of hTERT induces apoptosis independently of telomerase enzymatic activity in human breast cancer cells. Expression of a hTERT mutant lacking telomerase activity rescues the cells with lowered telomerase without inducing cell death. With similar patterns of subcellular distribution to that of the tumor suppressor protein p53 during mitosis, hTERT interacts with p53 and poly(ADP-ribose) polymerase (PARP). Decreasing p53 expression in intact cells worsens, and increasing p53 prevents, cell death induced by lowering hTERT. Thus, hTERT maintains cell survival and proliferation via both telomerase enzymatic activity-dependent telomere lengthening and enzymatic activity-independent intermolecular interactions involving p53 and PARP. PMID- 12082629 TI - Resistance of mitochondrial DNA-deficient cells to TRAIL: role of Bax in TRAIL induced apoptosis. AB - Mitochondrion is one of the master players in both apoptosis and necrosis. We studied the role of mitochondrial function in TRAIL-induced apoptosis. TRAIL killed SK-Hep1 cells with characteristic features of apoptosis such as DNA fragmentation, sub-G1 ploidy peak and cytochrome c translocation. In contrast, mitochondrial DNA-deficient SK-Hep1 rho(0) cells were resistant to TRAIL. Dissipation of mitochondrial potential or cytochrome c translocation did not occur in rho(0) cells after TRAIL treatment. TRAIL induced translocation of Bax subsequent to the cleavage of Bid in parental cells. However, Bax translocation was absent in rho(0) cells, accounting for the failure of cytochrome c release in rho(0) cells. Forced expression of Bax induced caspase-3 activity in rho(0) cells. Incubation of rho(0) cells with ADP+Pi to increase intracellular ATP restored sensitivity to TRAIL. Despite different sensitivity to TRAIL, parental cells and rho(0) cells did not show significant difference in susceptibility to agonistic anti-Fas antibody, TNF-alpha or staurosporine. Our results indicate that TRAIL-induced apoptosis is dependent on intact mitochondrial function and susceptibility of mitochondrial DNA-deficient cells to apoptosis depends on the type of apoptotic stimuli. Tumor cells with mitochondrial mutations or dysfunction might have the ability to evade tumor surveillance imposed by TRAIL in vivo. PMID- 12082630 TI - Adenovirus-mediated overexpression of p14(ARF) induces p53 and Bax-independent apoptosis. AB - The human INK4a gene locus encodes two structurally unrelated tumor suppressor proteins, p16(INK4a) and p14(ARF), which are frequently inactivated in human cancer. Whereas p16(INK4a) acts through engagement of the Rb-cdk4/6-cyclin D pathway, both the pro-apoptotic and cell cycle-regulatory functions of p14(ARF) were shown to be primarily dependent on the presence of functional p53. Recent reports have also implicated p14(ARF) in p53-independent mechanisms of cell cycle regulation and apoptosis induction, respectively. To further explore the pro apoptotic function of p14(ARF) in relation to functional cellular p53, we constructed a replication-deficient adenoviral vector for overexpression of p14(ARF) (Ad-p14(ARF)). As expected, Ad-p14(ARF) efficiently induced apoptosis in p53/Rb wild-type U-2OS osteosarcoma cells at low multiplicities of infection. Interestingly, Ad-p14(ARF) also induced apoptosis in both p53-deleted SAOS-2 osteosarcoma cells and HCT116 colon cancer cells with a bi-allelic knock-out of p53 (HCT116-p53(-/-)). Similarly, adenovirus-mediated overexpression of p14(ARF) induced apoptosis in p53/Bax-mutated DU145 prostate cancer cells as well as in HCT116 cells devoid of functional Bax (HCT116-Bax(-/-)). Restoration of Bax expression by retroviral gene transfer in DU145 cells did not further enhance p14(ARF)-triggered cell death. Infection with Ad-p14(ARF) induced activation of mitochondrial permeability shift transition, caspase activation and apoptotic DNA fragmentation irrespective of the presence or absence of either Bax or functional cellular p53. Nevertheless, overexpression of the anti-apoptotic Bcl-2 homolog Bcl-x(L) markedly inhibited p14(ARF)-induced apoptosis. This may indicate that p14(ARF) triggers a so far unknown activator of mitochondrial apoptosis which can be inhibited by Bcl-2 but which acts either independently or downstream of Bax. Taken together, this report demonstrates the participation of signaling pathways apart from the p53/Mdm-2 rheostat and Bax in p14(ARF)-mediated apoptosis. PMID- 12082631 TI - Downregulation of human polo-like kinase activity by antisense oligonucleotides induces growth inhibition in cancer cells. AB - A central role for polo-like kinases (PLK) in regulating several stages of mitotic progression has been born out in several species. Overexpression of PLK1 is observed in the majority of hitherto analysed human tumors. PLK1 overexpression is a negative prognostic factor in patients suffering from non small cell lung cancer, head and neck tumors, esophageal carcinomas and melanomas. In order to define the role of PLK1 for mitotic progression of human cells and for neoplastic cell growth, phosphorothioate antisense oligonucleotides (ASOs) were tested to selectively downregulate PLK1 expression in MDA-MB-435 (breast cancer), HeLa S3 (cervical carcinoma) and A549 (non-small cell lung cancer) cells. ASOs were identified which suppress PLK1 mRNA and protein in a dose-dependent and sequence-specific manner. This approach also led to reduced PLK1 serine/threonine kinase activity. Downregulation of cellular PLK1 levels in cancer cells altered cell cycle progression moderately with an elevated percentage (20-30%) of cells in G(2)/M. Furthermore, cells with reduced PLK1 protein gained a rounded phenotype with multiple centrosomes. Moreover, ASO treatment resulted in potent antiproliferative effects in cell culture. Considerable antitumor activity was observed in vivo against A549 cells. This study suggests that antisense inhibitors targeted against PLK1 at well tolerated doses may be considered as a cancer therapeutic agent. PMID- 12082632 TI - WISP3 is a novel tumor suppressor gene of inflammatory breast cancer. AB - Inflammatory breast cancer (IBC) is an aggressive form of breast cancer with a 5 year disease-free survival of less than 45%. Little is known about the genetic alterations that result in IBC. In our previous work, we found that WISP3 was specifically lost in human IBC tumors when compared to stage-matched, non-IBC tumors. We hypothesize that WISP3 has tumor suppressor function in the breast and that it may be a key genetic alteration that contributes to the unique IBC phenotype. The full-length WISP3 cDNA was sequenced and cloned into an expression vector. The resulting construct was introduced in to the SUM149 cell line that was derived from a patient with IBC and lacks WISP3 expression. In soft agar, stable WISP3 transfectants formed significantly fewer colonies than the controls. Stable WISP3 transfectants lost their ability to invade and had reduced angiogenic potential. WISP3 transfection was effective in suppressing in vivo tumor growth in nude mice. Mice bearing WISP3 expressing tumors had a significantly longer survival than those with vector-control transfectant tumors. Our data demonstrate that WISP3 acts as a tumor suppressor gene in the breast. Loss of WISP3 expression contributes to the phenotype of IBC by regulating tumor cell growth, invasion and angiogenesis. PMID- 12082633 TI - Proapoptotic activity of ITM2B(s), a BH3-only protein induced upon IL-2 deprivation which interacts with Bcl-2. AB - Growth factor deprivation is a physiological mechanism to induce apoptosis. We used an IL-2-dependent murine T cell line to identify proteins that trigger apoptosis. Here we report the identification, the cloning and characterization of ITM2B(s), a protein induced upon IL-2-deprivation. ITM2B(s), which shares the BH3 domain of Bcl-2 family members, is a cytoplasmic and mitochondrial protein. Expression of ITM2B(s) induces apoptosis in IL-2-stimulated cells, but not in IL 4-stimulated cells, while overexpression of the long form of the protein is not able to induce apoptosis. In IL-2-stimulated cells, ITM2B(s) interacts with the antiapoptotic protein Bcl-2, and does not interact with the proapoptotic Bad. Mutation of the critical L and D residues within the BH3 domain abolished the ability of ITM2B(s) to promote apoptosis. PMID- 12082634 TI - Overexpression of the HMGA2 gene in transgenic mice leads to the onset of pituitary adenomas. AB - Overexpression of the HMGA2 gene is a common feature of neoplastic cells both in experimental and human models. Intragenic and extragenic HMGA2 rearrangements responsible for HMGA2 gene overexpression have been frequently detected in human benign tumours of mesenchymal origin. To better understand the role of HMGA2 overexpression in human tumorigenesis, we have generated transgenic mice carrying the HMGA2 gene under the transcriptional control of the cytomegalovirus promoter. High expression of the transgene was demonstrated in all the mouse tissues analysed, whereas no expression of the endogenous HMGA2 gene was detected in the same tissues from wild-type mice. In this study, two independent lines of transgenic mice have been generated. By 6 months of age, 85% of female animals of both transgenic lines developed pituitary adenomas secreting prolactin and growth hormone. The transgenic males developed the same phenotype with a lower penetrance (40%) and a longer latency period (about 18 months). Therefore, these data demonstrate that the overexpression of HMGA2 leads to the onset of mixed growth hormone/prolactin cell pituitary adenomas. These transgenic mice may represent an important tool for the study of this kind of neoplasia. PMID- 12082635 TI - BRCA1 transactivates the cyclin-dependent kinase inhibitor p27(Kip1). AB - The p27(Kip1) is a member of the universal cyclin-dependent kinase inhibitor family. Previously, immunochemical analysis of a series of breast cancer cell lines demonstrated a correlation between the expression of p27(Kip1) and the breast cancer susceptibility gene BRCA1. BRCA1 has a number of activities including DNA repair, growth inhibition and as a transcription factor. Here we demonstrate that BRCA1 transactivates expression of p27(Kip1). This transactivation is dependent on the presence of a functional C-terminal transactivation domain. Promoter-deletion analysis identified the presence of a putative BRCA1-responsive element located at position -615 to -511 of the p27(Kip1) promoter. These results suggest that the transcriptional regulation of p27(Kip1) by BRCA1 may be a mechanism for BRCA1- induced growth inhibition. PMID- 12082636 TI - Human p14(ARF)-mediated cell cycle arrest strictly depends on intact p53 signaling pathways. AB - The tumor suppressor ARF is transcribed from the INK4a/ARF locus in partly overlapping reading frames with the CDK inhibitor p16(Ink4a). ARF is able to antagonize the MDM2-mediated ubiquitination and degradation of p53, leading to either cell cycle arrest or apoptosis, depending on the cellular context. However, recent data point to additional p53-independent functions of mouse p19(ARF). Little is known about the dependency of human p14(ARF) function on p53 and its downstream genes. Therefore, we analysed the mechanism of p14(ARF) induced cell cycle arrest in several human cell types. Wild-type HCT116 colon carcinoma cells (p53(+/+)p21(CIP1+/+) 14-3-3sigma(+/+)), but not p53(-/-) counterparts, underwent G(1) and G(2) cell cycle arrest following infection with a p14(ARF)-adenovirus. In p21(CIP1-/-) cells, p14(ARF) did not induce G(1) or G(2) arrest, while 14-3-3sigma(-/-) counterparts were mainly arrested in G(1), pointing to essential roles of p21(CIP1) in G(1) and G(2) arrest and cooperative roles of p21 and 14-3-3sigma in ARF-mediated G(2) arrest. Our data demonstrate a strict p53 and p21(CIP1) dependency of p14(ARF)-induced cell cycle arrest in human cells. PMID- 12082637 TI - Blocking NF-kappaB activation in Jurkat leukemic T cells converts the survival agent and tumor promoter PMA into an apoptotic effector. AB - The transcription factor NF-kappaB promotes cell survival. Using a variant of Jurkat leukemic T cells expressing IkappaB-alphaDeltaN, a super-repressor of NF kappaB activation we first show that the tumor promoter PMA could prevent Fas induced apoptosis via activation of NF-kappaB. Moreover, we demonstrate that in the absence of NF-kappaB activation, PMA became a strong inducer of apoptosis through stimulation of the upstream caspases 8 and 9 as well as of the effector caspase 3. A RNase-protection analysis showed that PMA stimulated the expression of several known anti-apoptotic genes (TRAF1, TRAF4, c-IAP-1, c-IAP-2, Bfl-1, Bcl xl). In the absence of NF-kappaB activation, these survival influences were strongly lowered revealing the apoptotic effect of PMA. These results suggest that NF-kappaB activation could be an important step in the tumor promoting effect of PMA. PMID- 12082638 TI - BCR/ABL P190 transgenic mice develop leukemia in the absence of Crkl. AB - The Bcr/Abl fusion protein directly causes chronic myelogenous leukemia and Philadelphia-chromosome positive acute lymphoblastic leukemia. Multiple independent studies have implicated Crkl, a small adapter protein, in transduction of oncogenic signals of Bcr/Abl and Crkl tyrosine-phosphorylation is used as a diagnostic tool for Philadelphia-positive leukemia. To evaluate the contribution of Crkl to this type of leukemia, we generated mutant mice that lack Crkl expression. We found that the overall survival of P190 BCR/ABL crkl-/- mice was comparable to that of genetically matched P190 BCR/ABL crkl +/+ mice. Both genotypes developed lymphoid lineage leukemia/lymphoma. Western blot analysis of /- and +/+ lymphomas showed that the related Crk protein was tyrosine phosphorylated and could be found complexed with Bcr-Abl P190. These data indicate that possible therapeutic approaches that target Crkl may be complicated by the presence of pathways that compensate for lack of Crkl function. PMID- 12082639 TI - The leukemia-associated transcription repressor AML1/MDS1/EVI1 requires CtBP to induce abnormal growth and differentiation of murine hematopoietic cells. AB - The leukemia-associated fusion gene AML1/MDS1/EVI1 (AME) encodes a chimeric transcription factor that results from the (3;21)(q26;q22) translocation. This translocation is observed in patients with therapy-related myelodysplastic syndrome (MDS), with chronic myelogenous leukemia during the blast crisis (CML BC), and with de novo or therapy-related acute myeloid leukemia (AML). AME is obtained by in-frame fusion of the AML1 and MDS1/EVI1 genes. We have previously shown that AME is a transcriptional repressor that induces leukemia in mice. In order to elucidate the role of AME in leukemic transformation, we investigated the interaction of AME with the transcription co-regulator CtBP1 and with members of the histone deacetylase (HDAC) family. In this report, we show that AME physically interacts in vivo with CtBP1 and HDAC1 and that these co-repressors require distinct regions of AME for interaction. By using reporter gene assays, we demonstrate that AME represses gene transcription by CtBP1-dependent and CtBP1 independent mechanisms. Finally, we show that the interaction between AME and CtBP1 is biologically important and is necessary for growth upregulation and abnormal differentiation of the murine hematopoietic precursor cell line 32Dc13 and of murine bone marrow progenitors. PMID- 12082640 TI - Correlation of Snail expression with histological grade and lymph node status in breast carcinomas. AB - Snail is a zinc finger transcription factor that triggers the epithelial mesenchymal transition (EMT) by directly repressing E-cadherin expression. Snail is required for mesoderm and neural crest formation during embryonic development and has recently been implicated in the EMT associated with tumour progression. In a series of human breast carcinomas, we have analysed the expression of Snail and that of molecules of the E-cadherin/catenin complexes. We have also correlated these data with the pathological features of the tumours. We show that Snail expression inversely correlates with the grade of differentiation of the tumours and that it is expressed in all the infiltrating ductal carcinomas (IDC) presenting lymph node metastases that were analysed. In addition, Snail is expressed in some dedifferentiated tumours with a negative nodal status. Considering that Snail is involved in the induction of the invasive and migratory phenotype in epithelial cells, these results indicate that it is also involved in the progression of breast ductal tumours, where it could additionally serve as a marker of the metastatic potential. PMID- 12082641 TI - AML1 stimulates G1 to S progression via its transactivation domain. AB - Inhibition of AML1-mediated transactivation potently slows G1 to S cell cycle progression. In Ba/F3 cells, activation of exogenous AML1 (RUNX1)-ER with 4 hydroxytamoxifen prevents inhibition of G1 progression mediated by CBFbeta-SMMHC, a CBF oncoprotein. We expressed three AML1-ER variants with CBFbeta-SMMHC in Ba/F3 cells. In these lines, CBFbeta-SMMHC expression is regulated by the zinc responsive metallothionein promoter. Deletion of 72 AML1 C-terminal residues, which includes a transrepression domain, did not alter the activity of AML1-ER, whereas further deletion of 98 residues, removing the most potent AML1 transactivation domain (TAD), prevented rescue of cell cycle inhibition. Notably, the two variants which did not stimulate G1 exacerbated CBFbeta-SMMHC-mediated cell cycle arrest, suggesting that they dominantly inhibit AML1 activities. In addition, the two variants which stimulated G1 also induced apoptosis in 5-15% of the cells, an effect consistent with excessive G1 stimulation. These observations indicate that AML1 activates transcription of one or more genes critical for the G1 to S transition via its C-terminal transactivation domain. Inactivation of AML in acute leukemia is expected to slow proliferation unless additional genetic alterations co-exist which accelerate G1. PMID- 12082642 TI - Gene expression differences between the microsatellite instability (MIN) and chromosomal instability (CIN) phenotypes in colorectal cancer revealed by high density cDNA array hybridization. AB - Two distinct pathways of tumorigenesis exist in sporadic colorectal cancer. The microsatellite instability pathway (MIN), which is characterized by widespread microsatellite instability due to aberrant mismatch repair machinery, accounts for 15% of all sporadic colorectal cancers. The chromosomal instability (CIN) phenotype, which accounts for 85% of sporadic colorectal cancers, is characterized by gross chromosomal lesions but the underlying mechanism remains unclear. We have addressed differences in gene expression between the MIN and CIN colorectal cancer phenotypes in vitro by the use of high density cDNA filters to compare gene expression patterns between MIN and CIN colorectal cancer cell-lines yielding a panel of 73 consistently differentially expressed genes. Nine of these genes were subjected to confirmatory analysis by independent methods, of which six were confirmed as being differentially expressed; PLK, RanBP2 and CCNA2 were overexpressed in CIN lines while BTF3, H2AZ and PTPD1 were overexpressed in MIN lines. These six genes are involved in diverse processes, such as maintenance of chromatin architecture, DNA-damage checkpoint and cell cycle regulation, which may contribute to the CIN and MIN phenotypes. PMID- 12082643 TI - Genetic isolates in East Asia: a study of linkage disequilibrium in the X chromosome. AB - The background linkage disequilibrium (LD) in genetic isolates is of great interest in human genetics. Although many empirical studies have evaluated the background LD in European isolates, such as the Finnish and Sardinians, few data from other regions, such as Asia, have been reported. To evaluate the extent of background LD in East Asian genetic isolates, we analyzed the X chromosome in the Japanese population and in four Mongolian populations (Khalkh, Khoton, Uriankhai, and Zakhchin), the demographic histories of which are quite different from one another. Fisher's exact test revealed that the Japanese and Khalkh, which are the expanded populations, had the same or a relatively higher level of LD than did the Finnish, European American, and Sardinian populations. In contrast, the Khoton, Uriankhai, and Zakhchin populations, which have kept their population size constant, had a higher background LD. These results were consistent with previous genetic anthropological studies in European isolates and indicate that the Japanese and Khalkh populations could be utilized in the fine mapping of both complex and monogenic diseases, whereas the Khoton, Uriankhai, and Zakhchin populations could play an important role in the initial mapping of complex disease genes. PMID- 12082644 TI - Analysis of mitochondrial DNA diversity in the aleuts of the commander islands and its implications for the genetic history of beringia. AB - The Aleuts are aboriginal inhabitants of the Aleutian archipelago, including Bering and Copper (Medny) Islands of the Commanders, and seem to be the survivors of the inhabitants of the southern belt of the Bering Land Bridge that connected Chukotka/Kamchatka and Alaska during the end of the Ice Age. Thirty mtDNA samples collected in the Commanders, as well as seven mtDNA samples from Sireniki Eskimos in Chukotka who belong to the Beringian-specific subhaplogroup D2, were studied through complete sequencing. This analysis has provided evidence that all 37 of these mtDNAs are closely related, since they share the founding haplotype for subhaplogroup D2. We also demonstrated that, unlike the Eskimos and Na-Dene, the Aleuts of the Commanders were founded by a single lineage of haplogroup D2, which had acquired the novel transversion mutation 8910A. The phylogeny of haplogroup D complete sequences showed that (1) the D2 root sequence type originated among the latest inhabitants of Beringia and (2) the Aleut 8910A sublineage of D2 is a part of larger radiation of rooted D2, which gave rise to D2a (Na-Dene), D2b (Aleut), and D2c (Eskimo) sublineages. The geographic specificity and remarkable intrinsic diversity of D2 lineages support the refugial hypothesis, which assumes that the founding population of Eskimo-Aleut originated in Beringan/southwestern Alaskan refugia during the early postglacial period, rather than having reached the shores of Alaska as the result of recent wave of migration from interior Siberia. PMID- 12082645 TI - Evolving Issues in Oncology: What is the 'Optimal' Hemoglobin Level? Proceedings of an international symposium. Seville, Spain, 2000. PMID- 12082646 TI - Current clinical practices for ovarian cancers. AB - Ovarian cancer is one of the most aggressive gynecologic cancers. It shows its symptoms late and is consequently often diagnosed at an advanced stage. The search for a more effective chemotherapy regimen, therefore, is of great importance. Since 1996, the combination of cisplatin and paclitaxel has been proven to prolong survival in comparison with older regimens containing cisplatin and cyclophosphamide. In addition, the introduction of carboplatin in combination with paclitaxel showed similar efficacy but preferable toxicity profiles when compared with cisplatin in combination with paclitaxel. Representative studies evaluating paclitaxel combination therapies as well as new trends in the treatment of ovarian cancer are summarized in this article. PMID- 12082647 TI - The impact of anemia and its treatment on patients with gynecologic malignancies. AB - Anemia is a frequent complication of cancer and its treatment. It often impairs the functional status of patients and results in decreased functional capacity and quality of life. Its etiologies are multiple, including chronic inflammation, hemorrhage, nutritional deficiencies, hemolysis, bone marrow suppression by chemotherapy, or infiltration by tumor. It can manifest as feelings of weariness, tiredness, muscular weakness, dysphoric mood, somnolence, or impaired cognitive functioning. In gynecologic patients, the incidence of anemia has been reported to be as high as 80% depending on chemotherapy regimen. Given the various consequences of a low hemoglobin level, the importance of increasing or maintaining hemoglobin levels and ameliorating the symptoms is apparent. Clinical studies have demonstrated that the administration of recombinant human erythropoietin (rHuEPO, epoetin alfa) is effective and safe in increasing hemoglobin levels and improving the overall quality of life in patients with gynecologic cancers undergoing chemotherapy. Therefore, epoetin alfa treatment should be considered in this patient population. PMID- 12082648 TI - The Italian gynecological consensus statement on the use of epoetin alfa in the management of anemia. AB - A Consensus Conference on the use of recombinant human erythropoietin (rHuEPO, epoetin alfa) in gynecologic tumors was held in Rome in March 1999, and an associated consensus paper has been published in Italian. The current paper updates several discussions that took place at the 1999 meeting concerning epoetin alfa treatment in breast, ovarian, and cervical cancers; the role of epoetin alfa in mobilizing progenitor hematopoietic cells; administration of epoetin alfa in combination with granulocyte colony-stimulating factor; and the effect of hemoglobin levels on outcome of radiation or chemoradiation treatment. PMID- 12082649 TI - High-dose chemotherapy in breast cancer: where are we now? AB - High-dose chemotherapy (HDCT) with peripheral blood stem cell support is often used in high-risk patients with breast cancer. Most investigators performing these procedures provide anecdotal experience that complete remissions occur more commonly with HDCT than standard therapy. However, prospective studies have not convincingly demonstrated an advantage of HDCT in either the adjuvant or metastatic disease setting. The emergence of newer agents, such as the taxanes and trastuzumab, and their incorporation into treatment regimens may create a new standard against which HDCT must be evaluated. In addition, the heterogeneity of breast cancer patients represents an important barrier that must be addressed in future evaluations of HDCT. New prospective, randomized, clinical trials of HDCT are needed, which will incorporate the newer anticancer agents in standard therapy and enroll patients according to multiple well-defined risk factors. Until these studies are conducted, many questions will remain unanswered concerning the role of HDCT in breast cancer. Given its encouraging results in other malignancies, there is ample evidence to continue evaluating HDCT in controlled clinical trials. PMID- 12082650 TI - Anemia and epoetin alfa in high-dose chemotherapy programs for breast cancer patients. AB - Breast cancer patients undergoing therapeutic regimens of high-dose chemotherapy (HDCT) with circulating progenitor cell support often develop anemia. As a general consideration in the transplantation setting, red blood cell transfusions are normally required in patients who have passed the myeloablative phase after HDCT. In the initial 30-day period immediately following HDCT, the number of red blood cell units that are transfused will usually depend on a number of factors, including whether the transplantation was allogeneic or autologous. Observations and results from clinical studies have shown that the establishment of normal erythropoiesis varies depending on the source of the transplanted cells, resulting in different transfusion requirements. Several studies support the use of recombinant human erythropoietin (rHuEPO, epoetin alfa) after HDCT to ameliorate anemia and reduce transfusion requirements. Studies have also shown that administration of epoetin alfa prior to the myeloablative phase is an effective method for reducing red blood cell transfusion requirements in breast cancer patients receiving HDCT. Epoetin alfa in combination with other cytokines has been shown to positively affect the mobilization phase of hematopoietic progenitor cells for autografting. Furthermore, treatment with epoetin alfa could prove useful in bone marrow transplant recipients who experience delayed anemia. Recent studies that have addressed these topics in breast cancer indicate that, when used in the appropriate setting, epoetin alfa may play a role as a tool to decrease the need for red blood cell transfusion in patients undergoing HDCT plus autologous circulating progenitor cell support. PMID- 12082651 TI - New treatment approaches for lung cancer and impact on survival. AB - Lung cancer is an aggressive disease that is the leading cause of death from cancer in both males and females. Non-small cell lung cancer accounts for approximately 75% of all cases of lung cancer, and of these about 75% have locally advanced or disseminated disease. Most patients diagnosed with non-small cell lung cancer do not survive more than 2 years. Although chemotherapy has been shown to control symptoms and improve quality of life, there remains no standard, optimal chemotherapy regimen for non-small cell lung cancer. Regardless of regimen and chemotherapy agents administered, clinical trials have demonstrated response rates of 20% to 30%, median survival times between 35 and 40 weeks, and 1-year survival rates of 20% to 25%. Because traditional chemotherapy regimens have not shown substantial promise, new strategies are being explored for the treatment of lung cancer, including overcoming drug resistance, the use of antimetastatic and antiangiogenesis drugs, drugs that target novel molecular markers, signal transduction modulators, gene therapy, and vaccines. There are few but exciting new developments that may signal a more promising future for patients with lung cancer. PMID- 12082652 TI - Challenges in treating hematologic malignancies. AB - During the past 40 years substantial progress has been made in the treatment of hematologic malignancies, particularly in some subgroups of patients. Today, cure is attainable for patients with Hodgkin's disease and a considerable proportion of patients with high-grade non-Hodgkin's lymphoma. Prognosis is improving in patients with acute promyelocytic leukemia and, to some extent, those with acute lymphoblastic and myeloid leukemias. However, the majority of patients who suffer from a hematologic malignancy live with incurable disease. In CLL, outside the setting of a clinical trial, it is advisable to postpone treatment until the manifestation of clinical symptoms. It is yet to be determined whether treatment strategies based on new prognostic parameters such as cytogenetics can change the course of disease. In indolent lymphomas, cure is not attainable for the vast majority of patients; the median survival of 9 to 10 years has remained unchanged for several decades. Nevertheless, there has been a dramatic change in therapeutic paradigms in the past few years. For the first time, with the use of new cytostatic drugs and recombinant monoclonal antibodies, it is possible to achieve molecular remissions. Whether this will translate into cure or prolonged survival is still to be determined. In Hodgkin's disease, which is curable when treated with radiotherapy, chemotherapy, or combined therapy, depending on the stage of disease, the focus of future studies must be on prevention of early relapse and on primary resistant disease, both of which present a very poor prognosis. Finally, regardless of underlying malignancy and prognosis, the preservation of quality of life is of major consideration in the setting of hematologic malignancies. PMID- 12082653 TI - The effects of anemia in hematologic malignancies: more than a symptom. AB - Anemia is a common complication in patients with hematologic malignancies, and is caused by a variety of mechanisms, including neoplastic cell infiltration into the bone marrow, hemolysis, nutritional deficiencies, and defects in erythropoiesis as a result of the disease itself or cytotoxic therapy. The anemia associated with multiple myeloma is caused by inadequate erythropoietin levels consequent to renal impairment and the effect of inflammatory cytokines. The degree of anemia can have prognostic importance, as is the case with multiple myeloma, or be a significant indicator of disease stage, as noted with chronic lymphocytic leukemia. Anemia results in fatigue, exhaustion, dizziness, headache, dyspnea, and decreased motivation, seriously affecting a patient's quality of life. Since anemia is so prevalent in hematologic malignancy patients, its treatment must be an integral part of disease management, to improve quality of life and to possibly increase potential survival. Clinical studies have shown that effectively treating anemia and increasing hemoglobin levels using recombinant human erythropoietin (rHuEPO, epoetin alfa) has a significant effect on transfusion requirements and quality of life. PMID- 12082654 TI - Anemia of hematologic malignancies: what are the treatment options? AB - Anemia is a common complication in patients with hematologic malignancies, with incidence rates ranging up to 63%. In myelodysplastic syndromes, anemia is an essential feature of the disease. The decrease in hemoglobin may lead to several symptoms such as fatigue, exhaustion, and impaired quality of life, and it may worsen prognosis. Before the introduction of recombinant human erythropoietin (rHuEPO, epoetin alfa), red blood cell transfusions were the traditional treatment for improvement of Hb levels. Transfusions, however, are associated with several adverse events and risks, have only transient effects, and have a limited capacity to ameliorate the symptoms of anemia. Epoetin alfa represents a physiologic treatment option, especially in the long-term treatment of cancer- and cancer treatment-associated anemia, and is well tolerated, with response rates as high as 80%. Epoetin alfa is less effective in the treatment of the anemia of myelodysplastic syndrome, but appears to be synergistic with granulocyte-colony stimulating factor. However, not every patient responds to epoetin alfa; to avoid unnecessary interventions and costs, predictors of response have been proposed. This article outlines the advantages and disadvantages of the two major treatment forms of anemia: transfusions and epoetin alfa. Representative studies on the efficacy of epoetin alfa in anemic patients with hematologic malignancies as well as models to predict response to epoetin alfa treatment are summarized. PMID- 12082655 TI - Age, anemia, and fatigue. AB - Many conditions that would not be considered normal in a younger population are routinely accepted in older people as a part of so-called "normal" aging. Among these conditions are many chronic and debilitating conditions such as chronic pain, insomnia, weakness, fatigue, and anemia. This article reviews current evidence regarding the relationships among age, fatigue, weakness, anemia, and erythropoiesis. Anemia in the elderly is important because it can lead to weakness, fatigue, limitations in activity, and may increase cardiovascular risk. Recent studies of the effect of erythropoietin in an aging population support the hypothesis that anemia is associated with pathologic factors and not with normal aging. While older individuals admitted to hospitals are more likely to be anemic, these same individuals have a bone marrow mass and numbers of cultured progenitor cells that are similar to that of the younger population; therefore, the predicted response to erythropoietin, and thus the function of the bone marrow and cellular progenitors, is maintained. Thus, we can conclude that anemia is a correctable pathologic finding in elderly people. A number of studies have shown a strong relationship between fatigue and anemia, but few studies investigate to what degree age is a factor in weakness and fatigue. In a study of 375 anemic cancer patients with a median age of 61 years, age as a covariate in multiple linear regression analysis failed to reach significance for most measures of function and quality of life (QOL), including measures of energy, activities, mental health, general cancer-related QOL, and overall QOL. Additional analysis suggests that other factors, including cancer progression, hemoglobin change, and baseline hemoglobin levels, are much more important in determining change in functional and quality-of-life scores. In another set of 2,000 cancer patients and 1,000 controls, cancer patients experienced significantly more fatigue compared with controls. There was no correlation between cancer patient age and fatigue, while in controls the cohort aged 65 or more reported more fatigue than did younger subjects. Finally, measurement of QOL in the general population demonstrated, for both the Short-Form 36 and Functional Assessment of Cancer Therapy - Anemia questionnaires, that age alone is not significantly correlated with QOL. We suggest that chronic conditions such as fatigue and anemia are no more "normal" in an aging population than in a general population, and that all patients with chronic conditions be adequately treated and counseled for their condition. PMID- 12082656 TI - Advances in quality of life measurements in oncology patients. AB - Accurate assessment of the quality of life (QOL) of patients can provide important clinical information to physicians, especially in the area of oncology. Changes in QOL are important indicators of the impact of a new cytotoxic therapy, can affect a patient's willingness to continue treatment, and may aid in defining response in the absence of quantifiable endpoints such as tumor regression. Because QOL is becoming an increasingly important aspect in the management of patients with malignant disease, it is vital that the instruments used to measure QOL are reliable and accurate. Assessment of QOL involves a multidimensional approach that includes physical, functional, social, and emotional well-being, and the most comprehensive instruments measure at least three of these domains. Instruments to measure QOL can be generic (eg, the Nottingham Health Profile), targeted toward specific illnesses (eg, Functional Assessment of Cancer Therapy - Lung), or be a combination of generic and targeted. Two of the most widely used examples of the combination, or hybrid, instruments are the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 Items and the Functional Assessment of Chronic Illness Therapy. A consequence of the increasing international collaboration in clinical trials has been the growing necessity for instruments that are valid across languages and cultures. To assure the continuing reliability and validity of QOL instruments in this regard, item response theory can be applied. Techniques such as item response theory may be used in the future to construct QOL item banks containing large sets of validated questions that represent various levels of QOL domains. As QOL becomes increasingly important in understanding and approaching the overall management of cancer patients, the tools available to clinicians and researchers to assess QOL will continue to evolve. While the instruments currently available provide reliable and valid measurement, further improvements in precision and application are anticipated. PMID- 12082657 TI - Epoetin alfa for protection of metabolic and exercise capacity in cancer patients. AB - A controlled clinical trial was conducted to evaluate the use of epoetin alfa in cachectic patients with solid tumors who were not receiving chemotherapy to determine if increasing hemoglobin (Hb) resulted in increased exercise capacity, metabolism, and energy efficiency during a maximum work load. The randomized, prospective study included 108 patients who received oral indomethacin 50 mg twice daily (n = 58; control group), or oral indomethacin 50 mg twice daily with epoetin alfa 4,000 to 10,000 IU by subcutaneous injection 3 times weekly (n = 50; study group). Patients randomized to the study group received epoetin alfa only when Hb decreased below 12.8 g/dL for men and 12.0 g/dL for women. Mean Hb levels in the study group were significantly (P <.0001) improved overall compared with the control group, with significant differences seen between groups after 2 to 4 months (P <.003), 6 to 8 months (P <.01), and 10 to 30 months (P <.01). Mean inflammatory variables including serum albumin, erythrocyte sedimentation rate, and C-reactive protein were significantly (P <.02) changed in the study group compared with the control group (ie, the control group had more inflammation). Significantly lower mean body weight (P <.05) and resting energy expenditure (P <.007) were recorded for patients in the control group versus the study group. The study group showed significantly greater mean exercise capacity (P <.0001), mean oxygen uptake (P <.01), mean CO(2) production (P <.009), and respiration (P <.03). These results demonstrate that early use of epoetin alfa prevents anemia in patients with progressive cancer who are not receiving chemotherapy. Normalization of Hb levels resulted in improved whole-body metabolism and energy efficiency, which is associated with greater exercise capacity and better daily quality of life. PMID- 12082658 TI - Epoetin alfa in patients not on chemotherapy - Canadian data. AB - A prospective open-label study was designed to evaluate the safety, efficacy, and impact on quality of life of recombinant human erythropoietin (rHuEPO, epoetin alfa) therapy for cancer-related anemia. Of the 401 patients enrolled at 34 centers from across Canada, a cohort of 183 patients did not receive chemotherapy during the 16-week study period. All patients received epoetin alfa 150 IU/kg subcutaneously 3 times per week. The dose was increased to 300 IU/kg if the hemoglobin level did not increase by at least 1.0 g/dL after 4 weeks. Epoetin alfa therapy significantly increased hemoglobin levels and reduced transfusion requirements. Moreover, epoetin alfa provided statistically significant and clinically meaningful improvements in quality of life as measured by the Functional Assessment of Cancer Therapy-Anemia and Linear Analog Scale Assessment (also known as Cancer Linear Analog Scale). Increases in hemoglobin were correlated significantly with improvements in quality of life as well as Eastern Cooperative Oncology Group performance status. Treatment with epoetin alfa was well tolerated. These results demonstrate that epoetin alfa therapy is effective and safe in cancer patients with anemia, regardless of whether they are or are not receiving chemotherapy. PMID- 12082659 TI - The use of epoetin alfa in chemotherapy patients: a consistent profile of efficacy and safety. AB - Epoetin alfa has been in use for over a decade to increase hemoglobin in patients with cancer who develop chemotherapy-associated anemia. Early placebo-controlled, randomized studies, as well as recent large, community-based trials in thousands of patients, have consistently shown recombinant human erythropoietin (rHuEPO, epoetin alfa) to be effective and safe in the treatment of chemotherapy associated anemia. Patients experienced an improved quality of life (QOL) related to the magnitude of the hemoglobin increase. As measured by a Linear Analog Scale Assessment, mean energy level, ability to do daily activities, and overall QOL improved significantly in patients who received epoetin alfa. The improvement in QOL was similar when anemia-specific instruments (Functional Assessment of Cancer Therapy-Anemia) were used. A large community-based trial has demonstrated that the more convenient once-weekly dosing schedule resulted in good efficacy and safety profiles. A recent double-blind placebo-controlled study that compared the efficacy of epoetin alfa versus placebo in patients receiving nonplatinum-based chemotherapy has confirmed epoetin alfa's efficacy, safety, and beneficial QOL effects. These findings should challenge current anemia management practices and encourage aggressive treatment of anemia in cancer patients receiving chemotherapy. PMID- 12082660 TI - The optimal hemoglobin level in the cancer patient. PMID- 12082661 TI - Expert testimony 'unnecessary' high risk pt. falls from bed. PMID- 12082662 TI - OR nurse relegated to 'dummy room' sues under ADA. Case on point: Brown v. Lester E. Cox Med. Ctrs., No. 01-1096 (8th Cir. 4/17/02) S.W.3d.-MO. PMID- 12082663 TI - CA: overuse of an injection site--abscesses result: inadmissibility of Dr.'s Hearsay statement saves day. PMID- 12082664 TI - RI: outpatient falls in hospital--has no expert: nurse expert's testimony required by court. PMID- 12082665 TI - Workers' comp claim 23 years after 'needle stick'. Case on point: Young v. Cross County Hosp., No. CA01-1208 (Ark.App. 05/01/02) S.W.3d-AR. PMID- 12082666 TI - Masturbatory guilt leading to severe depression and erectile dysfunction. PMID- 12082667 TI - Are our definitions of women's desire, arousal and sexual pain disorders too broad and our definition of orgasmic disorder too narrow? AB - Since each individual female sexual dysfunction is complex, it is necessary to subtype them in addition to dividing them into life-long or acquired disorder. The complexity of women's sexual arousal necessitates appreciation of a number of different types of arousal disorders that vary not only in etiology but also in management. The coexistence of sexual arousal and sexual desire, which develops during a sexual experience, explains the frequent comorbidity of arousal and desire disorders. Subtyping of hypoactive sexual desire disorder allows analysis of lack of receptivity and of any marked loss of the traditional markers of sexual desire over and beyond a normative lessening with relationship duration. Dyspareunia and vaginismus require further analysis prior to any definitive therapy. The definition of orgasmic disorder needs to include loss of orgasmic intensity and the possibility of coincident arousal disorder. PMID- 12082668 TI - Marriage consummated after 22 years: a case report. AB - This case report discusses a couple who presented with a 22-year unconsummated marriage, due to penetration phobia in the female partner. The couple did well with sex education and therapy. The case is discussed in its various aspects and with a cultural background. PMID- 12082669 TI - The motivation and mental health of sex workers. AB - Commercial sex work presents specific mental health concerns. We aimed to study motivation for sex work and mental health issues in a sample of such women. We contacted 55 consenting women through organized brothels and interviewed them using the Farley questionnaire and screening items for posttraumatic stress disorder (PTSD) and depression. Eighty-two percent of the women had arrived illegally and had been "trafficked." All but 2 were engaged voluntarily in sex work. Seventeen percent met criteria for PTSD, and 19% were likely to be clinically depressed. We present representative case histories. Availability of mental health treatment for workers in the sex industry could improve compliance with HIV prevention programs and enlarge options for women to leave the sex industry. We observed that stereotypes of sex workers as either always having histories of childhood abuse or as being always "happy hookers" were incorrect. PMID- 12082670 TI - The Female Sexual Distress Scale (FSDS): initial validation of a standardized scale for assessment of sexually related personal distress in women. AB - Recent consensus-based characterizations of female sexual dysfunction have emphasized personal distress as an essential component of their definition. To assist researchers and clinicians, we developed a new scale, the Female Sexual Distress Scale, to measure sexually related personal distress in women. In this article, we describe the initial stages in the development and validation of this instrument. Three studies involving a total of approximately 500 women were performed to evaluate the reliability and validity of the scale in different samples of sexually functional and dysfunctional women. Results indicated a unidimensional factor structure in both the original 20-item version and in a "polished" 12-item version. We observed a high degree of internal consistency and test-retest reliability in both versions across all three studies. Additionally, the scale showed a high degree of discriminative ability to distinguish between sexually dysfunctional and functional women in each of the studies. One study also showed a strong sensitivity to treatment response. Finally, we observed moderate positive correlations with other conceptually related nonsexual measures of distress, supporting the construct validity of the scale. Overall, these findings provide solid support for the FSDS as a valid and reliable measure for assessing sexually related personal distress in women. PMID- 12082671 TI - Reliability of retrospective self-reports of sexual and nonsexual health behaviors among women. AB - The accuracy of self-reports regarding sexual health behavior has been questioned. To investigate whether sexual health behaviors are uniquely difficult to report, we asked 185 college women to answer behavioral frequency questions about sexual and nonsexual health behaviors for an 8-week interval. Women either took part in a face-to-face interview or completed a self-administered questionnaire. One week later, the women returned and responded to the same questions in the same mode of assessment conditions. The test-retest intraclass correlations showed that all health behaviors, sexual and nonsexual, were reported reliably. There was a trend for lower-frequency reports to yield more stable estimates of behavioral frequency. These findings converge with other methodological investigations to indicate that socially sensitive health behaviors are not more difficult to assess reliably. PMID- 12082672 TI - The influence of a vasectomy on the marital relationship and sexual satisfaction of the married man. AB - The purpose of this study was to determine whether a vasectomy had any effect on important aspects of a marriage, such as sexual satisfaction, marital satisfaction, communication, and frequency of sexual intercourse. An experimental research design was used. We collected data by means of a biographical questionnaire, the Enriching & Nurturing Relationship Issues, Communications & Happiness questionnaire (Olson et al., 1985), and the Index of Sexual Satisfaction (Fischer & Corcoran, 1990), which were applied prior to and 5 months after the vasectomy. Results indicated no significant difference between the before and after measurements regarding sexual satisfaction, marital satisfaction, communication, and frequency of sexual intercourse. PMID- 12082673 TI - Sexual secrets, trauma, and dysfunction. AB - The majority of people who present for couple sex therapy have sexual secrets, often involving both past and present experiences. This article explores the most common sexual secrets, the ways that secrets can subvert treatment, the decision whether to share secrets, techniques for sharing secrets, and the motivating impact on marital trust and sexuality of confronting secrets. PMID- 12082674 TI - Parting gifts: termination rituals in group therapy. AB - A patient's termination from group therapy is a powerful experience for the departing patient, the therapist, and all group members. Unless the feelings evoked are channeled into constructive expression, they may undermine this potentially valuable phase of both the departing patient's group treatment and the life of the group as a whole. A termination ritual, styled by a particular patient according to his or her own need, therapy goals, and personality may help the patient achieve a more clearly defined sense of self. The authors suggest that the group therapist's careful attunement to and thorough exploration of the significance of any termination ritual or gift will help to extract maximum therapeutic benefit for the departing member and the group as a whole. PMID- 12082675 TI - Acting-out: its functions within analytic group psychotherapy and its transformation into dreams. AB - In group processes, acting-out has diverse functions, all of them equally important. It has an intrapsychic, interpersonal, and group dynamic function. Not only may it be understood as a form of resistance, but also in its communicative and reparative potential. The authors investigate the thesis that acting-out also contains the seed for change, thus helping patients divest themselves of pathological behavior. Using a group process as an example, this article shows how boundaries can be drawn between past and present experiences while using the communicative and reparative functions of acting-out. Unconscious psychodynamics can then be transformed from acting-out into dreams. PMID- 12082676 TI - Passion in group: thinking about loving, hating, and knowing. AB - In his early work, Bion (1961) established the goal of learning about and getting beyond the basic assumptions to become a work group. Later, in his structural theory of affect, passion became a key concept. Passion describes the necessary and sufficient condition for a psychotherapy group to be a work group. Passion is an intersubjective process of bearing and utilizing one's most basic affects to reach self-conscious emotional awareness. Bion postulated three primary affects: loving, hating, and knowing (LHK). A clinical example illustrates how the therapist may represent, mentally organize, and mobilize the group's potential for passion by attending to the evolution of his or her own affects. Passion transcends transference-countertransference in that an optimal level of personal meaning from LHK is achieved and utilized in emotional participation. PMID- 12082677 TI - Changing the guard: new leadership for an established group. AB - When the leader of an established therapy group decides to leave, the group may wish to continue with a new leader. This decision deserves careful exploration. The transfer of leadership to a new leader is a powerful event in a group. New leaders often experience strong countertransference reactions. Specific recommendations for new leaders include meeting individually with each continuing member, helping the group stabilize and process the meaning of the change, and developing ownership of the leader role. When managed successfully, a leadership transition can strengthen the group cohesion and offer important therapeutic opportunity. PMID- 12082678 TI - The ghost in the group room: countertransferential pressures associated with conjoint individual and group psychotherapy. AB - Conjoint individual and group therapy is a commonly used, effective psychotherapeutic treatment. The relationship between the conjoint therapists significantly influences the treatment. This article examines countertransference dilemmas of the group and individual therapists arising from the real and fantasied conjoint relationship. Case examples illustrate specific countertransference dilemmas. Therapists are encouraged to consider the possibility of difficulties with each potential conjoint treatment. Consultation and supervision are recommended to help the conjoint therapists identify and manage countertransference pressures. PMID- 12082679 TI - Comparison of group climate in an eating disorders partial hospital group and a psychiatric partial hospital group. AB - Women in an eating disorders partial hospital program and a psychiatric partial hospital program were compared on a self-report measure of group climate following a psychodynamic-interpersonal therapy group. Those with eating disorders experienced their groups as more engaged and as more avoiding than those in the psychiatric partial hospital group. Therapists may be able to use initial heightened engagement in eating disorder groups to counteract the tendency to avoid content. PMID- 12082680 TI - Change during and after long-term analytic group psychotherapy. AB - There are very few empirical studies of long-term, analytic group psychotherapy. Sixty-nine patients in long-term analytic group psychotherapy were evaluated from pretherapy to posttherapy, and one year after termination. The primary outcome measures were the Symptom Checklist-90 (Revised) and the Inventory of Interpersonal Problems. In addition, the Global Assessment of Functioning (GAF) was scored independently by two evaluators. The average time in treatment was 32.5 months. The patients improved significantly during treatment, up to 30 months, on all measures. The improvement continued during the follow-up period. Sixty to 86% of the patients were recovered or significantly changed at follow up. PMID- 12082681 TI - Responses to "The subjective experience of the self in the large group: two models for study". PMID- 12082682 TI - 3-D-conformal radiation therapy for pediatric giant cell tumors of the skull base. AB - BACKGROUND: Giant cell tumors (GCT) of the base of skull are rare neoplasms. This report reviews the treatment of four pediatric patients presenting with aggressive giant cell tumor, using fractionated and combined, conformal proton and photon radiation therapy at Massachusetts General Hospital and Harvard Cyclotron Laboratory. PATIENTS AND METHODS: Three female patients and one adolescent male, ages 10-15 years, had undergone prior, extensive surgical resection(s) and were treated for either primary (two patients) or recurrent (two patients) disease. Gross residual tumor was evident in three patients and microscopic disease suspected in one patient. Combined proton and photon radiation therapy was based on three-dimensional (3-D) planning, consisting of fractionated treatment, one fraction per day at 1.8 CGE (cobalt-gray equivalent) to total target doses of 57.6, 57.6, 59.4, and 61.2 Gy/CGE. RESULTS: With observation times of 3.1 years, 3.3, 5.3, and 5.8 years, all four patients were alive and well and remained locally controlled without evidence of recurrent disease. Except for one patient with partial pituitary insufficiency following radiotherapy for sellar recurrent disease, thus far no late effects attributable to radiation therapy have been observed. CONCLUSIONS: 3-D-conformal radiation therapy offers a realistic chance of tumor control for aggressive giant cell tumor in the skull base, either postoperatively or at time of recurrence. Conformal treatment techniques allow the safe delivery of relatively high radiation doses in the pediatric patient without apparent increase of side effects. PMID- 12082683 TI - Involved-field irradiation in combination with total-body irradiation (TBI) does not increase short-term toxicity compared to TBI alone in patients with advanced stage low-grade non-Hodgkin lymphoma. AB - PURPOSE: High-dose therapy (HDT) is currently under investigation for patients with advanced low-grade non-Hodgkin lymphoma (NHL). We report on the toxicity of a modified HDT that combines total-body irradiation (TBI) with involved-field irradiation (IF-RT) for patients with bulky disease or residual lymphomas > 2 cm after induction chemotherapy. PATIENTS AND METHODS: 41 patients received HDT which consisted of high-dose cyclophosphamide and fractionated TBI (6 x 2 Gy) followed by autologous stem cell transplantation. Eleven patients received IF-RT prior to TBI, three patients had already received another radiotherapy treatment prior to HDT. RESULTS: After a medium follow-up of 19 months we observed an overall survival rate of 100%, and a relapse-free survival rate of 78%. Severe toxicity was observed only in one patient who developed a myelodysplastic syndrome, and another patient who showed signs of pneumonitis. About two thirds of the patients claimed minor toxicity of grade I-II according the LENT-SOMA scale, predominantly as a decrease in concentration, reduced sexual functioning, and musculo-skeletal pain. Correspondingly, laboratory tests showed grade I-II changes of blood counts, liver enzymes, hormone levels, and lung function. There was no excess toxicity in the patients who received IF-RT additional to TBI. CONCLUSIONS: HDT including TBI and prior IF-RT is feasible without excess morbidity. Careful follow-up is required to detect myelodysplastic syndrome or endocrine changes of ovarian or testicular function. PMID- 12082684 TI - [Assessment of cognitive function after preventive and therapeutic whole brain irradiation using neuropsychological testing]. AB - PURPOSE: Aim of this study was the assessment of neuropsychological changes after whole brain irradiation. PATIENTS AND METHOD: 64 patients were tested before, and 29 after whole brain irradiation, including 28 patients with small cell lung cancer (SCLC) before prophylactic cranial irradiation (PCI) and 36 patients with cerebral metastases before therapeutic cranial irradiation (TCI), as well as 14 patients after PCI and 15 after TCI (Table 1). Intelligence, attention and memory were assessed applying a 90-minute test battery of standardized, neuropsychological tests (Table 3). RESULTS: Patients with SCLC showed test results significantly below average before PCI (n = 28, mean IQ = 83, SD = 17). Neither after PCI, nor after TCI the tested neuropsychological functions decreased significantly (Tables 4, 5). A comparison between SCLC-patients with and without cerebral metastases before whole brain irradiation showed better test results in patients with cerebral metastases and fewer cycles of preceding chemotherapy (Table 7). CONCLUSION: Neuropsychological capacity in patients with SCLC was impaired even before PCI. Possible reason is the preceding chemotherapy. Whole brain irradiation did not induce a significant decline of cognitive functions in patients with PCI or TCI. A decline in a longer follow-up nevertheless seems possible. PMID- 12082685 TI - Impact of the "belly board" device on treatment reproducibility in preoperative radiotherapy for rectal cancer. AB - BACKGROUND: The use of the belly board device (BBD) in the prone position has gained acceptance to spare small bowel in rectal cancer patients irradiated postoperatively, but there are few data in the preoperative setting, and the advantages of the BBD regarding normal tissue sparing may be counteracted by problems of patient positioning. This study was undertaken to investigate prospectively the influence of the BBD on treatment reproducibility in patients irradiated preoperatively in the prone position. PATIENTS AND METHODS: 23 patients with rectal carcinoma in clinical stages II/III were included in this study. Axis displacement was evaluated in 14 patients treated without the BBD and nine with. The BBD is a commercial device (Belly Board, Radiation Products Design, Albertville, MN) made of a 17-cm thick hard sponge with an opening of 42 x 42 cm2. No specific patient immobilization devices were used. During radiotherapy, twelve patients had four control films, while eleven patients had three. The mean treatment position deviation was calculated for the medio lateral, cranio-caudal and antero-posterior directions. RESULTS: When comparing the first control film to the corresponding simulation film for patients without the BBD and with the BBD, the mean lateral displacements were 1.5 mm and 3.2 mm (p = 0.26), the mean cranio-caudal displacements were 1.55 mm and 4.2 mm (p = 0.13), and the mean antero-posterior displacements were 1.8 mm and 4.5 mm (p = 0.04), respectively. When considering all control films, for the three directions, the amplitudes of the displacements were greater when using the BBD, particularly for the antero-posterior direction where the difference was highly significant (p = 0.0006). CONCLUSIONS: Our data show that, in patients treated prone for rectal cancer, the use of the BBD in the preoperative setting without immobilization devices was associated with problems of patient position reproducibility, particularly for the antero-posterior direction. Thus, the use of patient immobilization devices and/or individual custom-made BBD may be recommended if a decision to treat the patient with a BBD is taken. PMID- 12082686 TI - Interobserver comparison of CT and MRI-based prostate apex definition. Clinical relevance for conformal radiotherapy treatment planning. AB - BACKGROUND: CT is widely used for conformal radiotherapy treatment planning of prostate carcinoma. Its limitations are especially at the prostatic apex which cannot be separated from the urogenital diaphragm. The aim of this study was to compare the localization of the prostatic apex in CT and axial MRI to the sagittal MRI in an interobserver analysis. PATIENTS AND METHODS: 22 patients with pathologically proven prostatic carcinoma were included in the analysis. In all patients sagittal and axial T2-weighted MRI and conventional CT were performed. The position of the MRI and CT apices were localized independently by three observers in relation to the intertrochanteric line. Additional subjective judgment of the ability to define the apical border of the prostatic gland was performed by a five-scaled score. RESULTS: The apex of the prostate could be discriminated statistically significant (p < 0.001) better in the MRI as compared to CT with best judgement for the sagittal MRI. The interobserver variation for the definition of the prostatic apex was statistically significant (p = 0.009) smaller for the sagittal MRI compared to axial MRI and CT. On the average the apex as determined by sagittal MRI, axial MRI and CT was located 29 mm, 27 mm and 24 mm above the intertrochanteric line. The apex defined by CT would have led to an additional treatment of 6-13 mm in 10/22 patients compared to the sagittal MRI, defined by axial MRI only in five patients. CONCLUSION: Additional MRI provides a superior anatomic information especially in the apical portion of the prostate. It should be recommended for every single patient in the treatment planning process. It helps to avoid an unnecessary irradiation of healthy tissue and could lead to a decrease of anal side effects and radiation-induced impotency due to a reduction of the extent of irradiated penile structures. PMID- 12082687 TI - Transient DNA double-strand breakage in 4-hydroperoxyifosfamide-treated mammalian cells in vitro does not interact with the rejoining of radiation-induced double strand breaks. AB - BACKGROUND: 4-Hydroxyifosfamide is the primary metabolite in vivo of the bifunctional alkylating cytostatic ifosfamide. DNA interstrand cross-linking induced by bifunctional alkylators may be repaired through an intermediate with unligated repair patches on both strands which should uncover analytically as DNA double-strand breaks and allow to measure the rejoining kinetic of this repair intermediate. Additionally, the combined effects of drug and radiation treatment on rejoining of double-strand breaks was investigated with two different mammalian cell lines. MATERIAL AND METHODS: V79 (rodent fibroblasts) and Widr (human colon carcinoma) cells were treated for 2 hours with 4 hydroperoxyifosfamide which rapidly decays to 4-hydoxyifosfamide in aqueous solution or were exposed in combination with ionizing radiation followed by incubation for repair with or without the drug. DNA double-strand breakage was measured by pulsed-field electrophoresis. RESULTS: The 2 hours 4 hydroperoxyifosfamide treatment (30 micrograms/ml) resulted in a pronounced DNA fragmentation that, 2-4 hours after drug removal, declined with an estimated half live of about 4 hours for both cell lines. When the cells were additionally irradiated with 10 Gy given in the middle of drug exposure, the residual fragmentation after 12 or 24 hours incubation for repair was only marginally increased, roughly corresponding to the respective value after radiation, alone. A continuous drug exposure of 6 hours (at 10 micrograms/ml) resulted in a fragmentation that was independent of a preirradiation with a high dose of 30 Gy, immediately before drug addition. CONCLUSIONS: The present data support the idea that unligated/unrejoined double-stranded DNA ends are generated during the repair of lesions from bifunctional alkylators. The rate of subsequent rejoining is in the order of magnitude of the slow rejoining of radiation-induced double strand breaks. Processing of double-stranded DNA damage from either 4 hydroperoxyifosfamid or radiation exposure is apparently unaffected in combined treatments. PMID- 12082688 TI - Systemic mast cell disease (SMCD) and bone pain. A case treated with radiotherapy. AB - BACKGROUND: Systemic mast cell disease (SMCD) is a rare disease characterized by a multitopic proliferation of cytologically and/or functionally abnormal tissue mast cells. SMCD preferentially involves the skin, spleen, liver, lymph nodes and the bone marrow. The cause of SMCD is unknown. Bony pain, caused by mast cell infiltration of the marrow cavity, is present in up to 28% of cases and is frequently chronic and difficult to palliate with medical therapy. CASE REPORT: We report one case of refractory bone pain in a 54-year-old female Caucasian patient with advanced SMCD and associated bony involvement, which was treated with radiotherapy for pain palliation. Between 1995 and 1998, the patient was irradiated at four different locations: 1) right shoulder and proximal right humerus, 2) both hands, 3) both knees, 4) left humerus with a total dose of 40 Gy in 2.0 or 2.5 Gy daily fractions. RESULTS: Different results of pain palliation were achieved. In one location the pain was reduced for 55 months until her death due to disease progression, whereas in two other locations a pain control was maintained for 3 and 6 months after radiotherapy. In one location, no pain reduction was achieved. Severe side effects were not observed. CONCLUSION: Palliative radiotherapy has a role in the control of severe intractable bone pain in patients with advanced SMCD, though in some cases the effect may be short or incomplete. The observed palliation of pain can even differ in the same patient. PMID- 12082689 TI - [Radiotherapy within the scope of allogeneic kidney transplantation: an indication for local irradiation?]. AB - PURPOSE: In the past decades the indications for local graft irradiation (LGI) in acute renal transplant rejection have been limited and considered unfavorably. Despite major advantages in maintenance immunosuppression and management of acute allograft rejection a minority of patients remains with drug resistant transplant rejection. This subgroup of patients may benefit from LGI. PATIENTS AND METHODS: Between 1979 and 1990, eight patients with biopsy-proven acute renal allograft rejection and failure of all other immunosuppressive measures (corticosteroids, ATG, ALG or OKT3) were treated with LGI. Retrospective analysis was conducted for this control group. Radiotherapy was performed with Co-60 up to a median total dose of 6.0 Gy (single doses: 1.5-2.0 Gy). Six of eight patients were dialysis dependent prior to irradiation. In addition a literature review was performed including most important textbooks, electronic databases (Medline, Embase, Science Citations Index), and the internet. RESULTS: Two of eight patients experienced a clinical reversal of rejection and an improvement of renal function: serum creatinine decreased significantly. One patient remained free of dialysis with a functioning graft, the other had a recurrent rejection 2 months later and became dialysis dependent. The literature review showed, that adjuvant LGI has no advantage over conventional immunosuppression. However, in case of a drug refractory allograft rejection LGI restores long-term stable organ function in 13-60% of cases. CONCLUSION: The value of LGI of organ transplants, like renal allografts, is still not clearly defined. As a rescue measure in drug refractory allograft rejection special patients may clinically benefit, when a transplant nephrectomy can be avoided. Further prospective clinical trials are needed for a better assessment of LGI in organ transplantation. PMID- 12082690 TI - [Control of patients in an outpatient clinic of rheumatology]. AB - BACKGROUND: A high number of patients attending outpatient clinics of rheumatology are control cases. This study aimed at investigating the causes of such controls and how necessary they are. MATERIAL AND METHODS: 400 consecutive control patients selected from a local hospital were included. RESULTS: Inflammatory rheumatic diseases constituted 89% of the controls; rheumatoid arthritis was the diagnosis in 28% of cases. Routine control of patients with chronic disease was the main cause of control (41%). 15% of controls were regarded as partly or completely unnecessary; 95% of these were not repeated. INTERPRETATION: The reasons for controlling patients in rheumatology should be better defined. Regular evaluation could result in improved service to outpatients. PMID- 12082691 TI - [Melatonin treatment of a blind child with serious sleep disorders]. AB - BACKGROUND: Children with developmental and neurological disabilities are prone to develop serious sleep-wake cycle disorders that may be difficult to treat. MATERIAL AND METHODS: Case history. RESULTS: A 5-year old blind boy with multiple disabilities developed a chronic sleep-wake cycle disorder as his main clinical problem. Treatment included introduction of strict sleep habits and strengthening of environmental "zeitgebers". After five months melatonin 3 mg was administered at night for 4 weeks. The observation period also included 3 weeks without melatonin. Sleep was registered prospectively by a sleep diary. Strict sleep habits combined with strengthening of "zeitgebers" partially improved the sleep problems, but did not establish a normal sleep pattern. When melatonin was added, he normalized his sleep pattern in a few days. His sleep problems returned during the weeks in which he did not receive melatonin. No side effects were observed. INTERPRETATION: Melatonin is a promising treatment alternative for serious sleep problems in blind children. PMID- 12082692 TI - [Melatonin treatment of sleep disorders in disabled children]. AB - BACKGROUND: There is a high prevalence of chronic sleep-wake cycle disorders in developmentally and neurologically disabled children. Such disorders are often resistant to hypnotic and psychotropic drugs. Administration of exogenous melatonin can have an effect upon the circadian rhythm and establish a normal sleep-wake cycle. MATERIAL AND METHODS: Medline, Premedline and EBM Reviews were searched for articles on melatonin treatment of sleep-wake cycle disorders in children. Additionally, sleep was registered systematically in 15 developmentally or neurologically disabled children treated with melatonin. RESULTS AND INTERPRETATION: The search identified 75 articles of which 33 included neurologically or developmentally disabled children. Reported results generally indicate that melatonin improves sleep-wake cycle disorders in these children. However, the majority of studies were open, and knowledge about side effects and long-term effects is limited. Our own experience is in accordance with the literature; melatonin had positive effects in 13 children. Suggestions for practical use of melatonin in disabled children are presented. PMID- 12082693 TI - [Warfarin treatment of venous thromboembolism]. AB - BACKGROUND: Warfarin treatment of venous thromboembolism is the most frequent cause of reported serious and fatal adverse events associated with drug therapy in Norway. We assessed quality of treatment during transfer from hospital to community-based care. MATERIAL AND METHODS: 66 out of 100 consecutive patients with venous thromboembolism were studied by a retrospective survey that included data from hospital records and a questionnaire survey. RESULTS: Time in therapeutic range was 57% during the first four weeks. Undertreatment was the most frequent deviation. Some patients reported a long time lag from INR measurement to dose prescription in community care; 42% did not receive written treatment information. There were six recurrences, but no serious or fatal bleeding complications during one year of observation. Patient satisfaction with information and treatment organisation was high. INTERPRETATION: There is room for improvement of patient information and treatment quality in outpatient care in our area. PMID- 12082694 TI - [Prevalence of hypothyroidism and hyperthyroidism in Nord-Trondelag]. AB - BACKGROUND: The objective of this survey was to examine the prevalence of thyroid disease including autoimmunity in Norway. MATERIALS AND METHODS: All 94,009 inhabitants aged 20 and above in Nord-Trondelag county, Norway were invited to participate in a health survey. RESULTS: The prevalence of formerly diagnosed hyperthyroidism was 2.5% in women and 0.6% in men, hypothyroidism 4.8% and 0.9% respectively, increasing with age. In individuals without a history of thyroid disease the median, 2.5 and 97.5 percentiles for TSH (mU/l) were 1.8 and 0.49-5.7 for women and 1.5 and 0.56-4.6 for men. When excluding individuals with positive thyroid peroxidase antibodies (TPOAb), the 97.5 percentiles were 3.6 mU/l for women and 3.4 mU/l for men. The prevalence of pathological TSH values in women and men were TSH > or = 10 mu/l 0.9% and 0.4%; TSH 4.1-9.9 mU/l 5.1% and 3.7%; TSH < or = 0.05 mU/l 0.5% and 0.2%. The prevalence of positive TPOAb was 13.9% in women and 2.8% in men, the lowest percentage was seen with TSH 0.2-2.0 mU/l and increased both with higher and lower TSH values and was higher in women except for those with TSH > or = 10 mU/l (85% TPOAb positive). CONCLUSION: In spite of a high prevalence of recognised thyroid disease in the population a considerable number have undiagnosed thyroid dysfunction and positive TPOAb. PMID- 12082695 TI - [Thyrotoxic periodic paralysis--an unusual complication of hyperthyroidism]. AB - BACKGROUND: Thyrotoxic periodic paralysis (TPP) is a complication of hyperthyroidism. MATERIALS AND METHODS: We describe two patients with TPP. RESULTS: A 26-year-old man from Vietnam had weight loss, tachycardia, palpitations and heat intolerance for five months. Episodic leg and arm weakness developed three months after debut of symptoms. The second patient, a 23-year old woman from the Philippines, had had episodic leg weakness in the evenings after dinner for three weeks. Her attacks resolved spontaneously overnight. Physical examination of both patients revealed tachycardia and symmetrical proximal weakness involving both arms and legs. ECG and electrolyte analysis indicated a severe hypokalaemia; thyroid function tests showed hyperthyroidism. Both patients were diagnosed as having Graves' thyrotoxicosis and TPP. They were initially treated with propranolol and subsequently with carbimazole. The first patient had recurrence of thyrotoxicosis and paralysis after 16 months, whereas the second patient has remained symptom-free. INTERPRETATION: TPP is most common in Asian males, very few cases are reported in females. In Western countries TPP is rare, but with increasing immigration, TPP is likely to occur more frequently. PMID- 12082696 TI - [Health and patient rights education at the University of Tromso]. PMID- 12082697 TI - [Vitamin D deficiency in children with celiac disease]. PMID- 12082698 TI - [Tsetse flies in Eastern Africa]. PMID- 12082699 TI - [Profit optimization and generic substitution]. PMID- 12082700 TI - [Instruction by the church and terminal care]. PMID- 12082701 TI - [Was Jesus dead after the crucifixion?]. PMID- 12082702 TI - [Can physicians' work in hospitals be better organized?]. PMID- 12082703 TI - [Military attacks against ambulances and health personnel]. PMID- 12082704 TI - [A bump on the neck]. PMID- 12082705 TI - [Quality and independence]. PMID- 12082706 TI - [A new myocardial infarction "epidemics"--fact or fiction?]. PMID- 12082708 TI - [Neurosurgery and investments]. PMID- 12082707 TI - [Anticoagulant therapy with warfarin--still difficult]. PMID- 12082709 TI - [Treatment of hydrocephalus with endoscopic third ventriculocisternostomy]. AB - BACKGROUND: Endoscopic third ventriculocisternostomy has gained widespread use in the treatment of hydrocephalus. MATERIALS AND METHODS: In 1999 and 2000 endoscopic third ventriculostomy was performed in 120 patients (60 males) at Rikshospitalet, Oslo, Norway. Median age was 3.3 years (range 0.0 to 71.7). 69 procedures were done as primary treatment of hydrocephalus and 51 procedures for shunt failure. RESULTS: After one year, 62% of the patients older than 6 months at time of treatment and 8% of the patients younger than 6 months were successfully treated for their hydrocephalus with ventriculostomy only. The success rate for patients with hydrocephalus secondary to aqueductal stenosis or a space-occupying lesion in the posterior fossa was 72% and 31% for patients with other causes of hydrocephalus. The success rate of endoscopic third ventriculostomy as primary treatment of hydrocephalus was 55%; for the procedure at time of shunt failure 45%. The overall likelihood of success was significantly correlated to age, cause of hydrocephalus, and whether the procedure was done as primary treatment of hydrocephalus or at time of shunt failure. INTERPRETATION: Endoscopic third ventriculostomy seems to be effective in treating hydrocephalus caused by aqueductal stenosis or a space-occupying lesion in the posterior fossa in patients older than 6 months. The procedure should not be routinely attempted on patients younger than 6 months. PMID- 12082711 TI - Cultural diversity and health care settings. PMID- 12082710 TI - [Severe falciparum malaria treated with exchange transfusion]. AB - BACKGROUND: Acute severe malaria can be seen at any Norwegian hospital. The prognosis of this serious disease depends on rapid and effective treatment. MATERIAL AND METHODS: Two case reports of patients treated with exchange transfusions are presented. RESULTS: Two young Norwegian women travelled without adequate prophylaxis against malaria in South-East Asia and Central and Southern Africa respectively. A few days after leaving the endemic areas they got high fever, joint and muscular pain, and headache. In one of the patients the symptoms were interpreted as flu symptoms; one week later the patient was admitted to hospital suffering from severe malaria. The other patient was admitted to hospital on the day she returned to Norway, six days after getting febrile. Thin blood films showed Plasmodium falciparum, with 30% and 40-50% parasitaemia respectively. Both patients were icteric with thrombocytopenia and increased creatinine. The first patient had severe renal failure and signs of cerebral affection. She was treated with haemodialysis and exchange transfusions. The other patient was treated with exchange transfusions because of her high parasitaemia. Both patients made a complete recovery and were discharged after three and two weeks. INTERPRETATION: Severe falciparum malaria is a condition with high case-fatality if diagnostics and treatment is not optimal. In especially severe cases, exchange transfusion may be lifesaving. PMID- 12082712 TI - Recommended practices for the LASIK procedure: sterile versus surgically clean technique. PMID- 12082713 TI - "Rubbing the sleep from my eyes...". How I (accidentally) became an ophthalmic nurse. PMID- 12082714 TI - Seeing the future of contacts through amethyst eyes. PMID- 12082715 TI - Confusion assessment method (CAM). PMID- 12082716 TI - Refractive surgery: an overview. AB - The status of refractive surgery is a dynamic and exciting field in ophthalmology. Current procedures are effective, safe and well tolerated. The future of refractive surgery continues to evolve, developing new and exhilarating forms of refractive correction. It remains to be seen which procedure will prove the most successful overall; or perhaps a variety of procedures will ultimately be the solution to the many facets of individual patient needs and desires. PMID- 12082717 TI - [History of nephrology in the past 100 years and the future prospects]. PMID- 12082718 TI - [History of nephrology in the past 100 years: Kidney function tests]. PMID- 12082719 TI - [History of nephrology in the past 100 years: Classification of nephritis]. PMID- 12082720 TI - [History of nephrology in the past 100 years: Transcutaneous kidney biopsy]. PMID- 12082721 TI - [History of nephrology in the past 100 years: Renovascular hypertension]. PMID- 12082722 TI - [History of nephrology in the past 100 years: Diabetic nephropathies]. PMID- 12082723 TI - [History of nephrology in the past 100 years: Significance of hyperfiltration theory in the nephrology]. PMID- 12082724 TI - [History of nephrology in the past 100 years: Kidney transplantation]. PMID- 12082725 TI - [History of nephrology in the past 100 years: Dialysis]. PMID- 12082726 TI - [History of nephrology in the past 100 years: Pathogenic genes of kidney diseases]. PMID- 12082727 TI - [History of nephrology in the past 100 years in Japan]. PMID- 12082728 TI - [History of nephrology in the past 100 years: Japanese contribution to development of nephritis models]. PMID- 12082729 TI - [History of nephrology in the past 100 years: Japanese contribution to studies on the renin-angiotensin system]. PMID- 12082730 TI - [Contribution of Japanese researchers to progress in studies on nephrology in the past 100 years: Beta2-microglobulin as an etiological factor of amyloidosis in patients undergoing hemodialysis]. PMID- 12082731 TI - [Contribution of Japanese researchers to progress in studies on nephrology in the past 100 years: Molecular cloning of water channels in the kidney collecting duct and the etiology of nephrogenic diabetes insipidus]. PMID- 12082732 TI - [Contribution of Japanese researchers to progress in studies on nephrology in the last 100 years: Molecular mechanism of calcium and vitamin D metabolism in the kidneys]. PMID- 12082733 TI - [Contribution of Japanese researchers to progress in studies on nephrology in the past 100 years: Lipoportein glomerulopathy]. PMID- 12082734 TI - [History of nephrology in the past 100 years: acute kidney failure]. PMID- 12082735 TI - [History of nephrology in the past 100 years: Chronic kidney failure]. PMID- 12082736 TI - [Progress in the therapy of chronic kidney failure in Japan: Dialysis and renal transplantation]. PMID- 12082737 TI - [History of nephrology in the last 100 years: Nephrotic syndrome]. PMID- 12082738 TI - [History of nephrology in the last 100 years: Acute rapidly progressive glomerulonephritis]. PMID- 12082739 TI - [History of nephrology in the last 100 years: Chronic glomerulonephritis--special reference to IgA nephropathy]. PMID- 12082740 TI - [History of nephrology in the last 100 years: Diabetic nephropathies]. PMID- 12082741 TI - [History of nephrology in the past 100 years: Collagen disease and vasculitis]. PMID- 12082742 TI - [History of nephrology in the past 100 years: Kidney and hypertension]. PMID- 12082743 TI - [History of nephrology in the past 100 years: Water-electrolyte imbalance]. PMID- 12082744 TI - [History of nephrology in the past 100 years: Toxic nephropathies]. PMID- 12082745 TI - [Retrospective view of nephrology in the 20th century and prospects for further progress in the 21 century(discussion)]. PMID- 12082746 TI - [Clinical guideline review: IgA nephropathies]. PMID- 12082747 TI - [Clinical guideline review: Diabetic nephropathies]. PMID- 12082748 TI - [Clinical guideline review: Guidelines for life style and diet in kidney disease]. PMID- 12082749 TI - [Clinical guideline review: Standards for initiation of chronic dialysis]. PMID- 12082750 TI - [Clinical guideline review: Rapidly progressive glomerulonephritis]. PMID- 12082751 TI - [WDHA syndrome by composite pheochromocytoma and ganglioneuroma]. PMID- 12082752 TI - [Hyperammonemia due to portal vein obstruction caused by multiple liver cysts]. PMID- 12082753 TI - [Two cases of lead poisoning with abdominal pain]. PMID- 12082754 TI - [Limb-shaking transient ischemic attack in internal carotid artery occlusion]. PMID- 12082755 TI - [Successful treatment of chronic myelogenous leukemia with extramedullary megakaryoblastic tumor by allogeneic bone marrow transplantation (from an unrelated donor)]. PMID- 12082756 TI - [Toxic shock syndrome caused by enterotoxin B]. PMID- 12082757 TI - [Research is--also--leadership]. PMID- 12082758 TI - [How do we find the patients with renal artery stenosis and hypertension who benefit from percutaneous transluminal angioplasty?]. PMID- 12082759 TI - [Treatment of renal artery stenosis]. AB - The attitude to treatment of renal artery stenosis has recently been modified from an active to a more expectant strategy based on informations from randomised studies. The primary treatment should be antihypertensive agents. Revascularisation should be considered in patients with refractory hypertension, recurrent pulmonary oedema, bilateral renal artery stenosis or progressive azotaemia, and in patients with a narrow stenosis to a single kidney. The treatment, i.e. surgery or PTA with or without stent, should be selected on an overall view of the patients' health using a combination of clinical, pathophysiological, and angiographic investigations. PMID- 12082760 TI - Growth factors controlling angiogenesis and lymphangiogenesis. PMID- 12082761 TI - [Osteoarthritis. Pathogenesis, clinical features and treatment]. AB - Osteoarthritis (OA) is the most common joint disease and one of the most common diseases altogether. Everybody above 60 years of age has pathological features of OA in at least one joint. Loss of articular cartilage is a crucial event in OA. Concomitant features are osseous deformation and sclerosing, shrinkage of the capsule, atrophy of muscles, and variable degrees of synovitis. The cardinal symptom of OA is pain. Later, deformation and malalignment with deprived function of the joint are seen. The clinical and radiological features make up the diagnosis. Patients with all grades of OA should be given information and physical therapy. Paracetamol is the first drug of choice. If paracetamol is insufficient to achieve pain control or inflammation is present, COX-2 specific inhibitors should be considered. If effusion is present, an intra-articular injection of long-acting steroid may be administered. Severe OA of the hip or knee is treated with arthroplastic surgery, but precise indications for surgery need to be established. PMID- 12082762 TI - [Photographic screening for diabetic retinopathy in the county of North Jutland. The first fully digitalized telemedicine-screening clinic]. AB - Screening for diabetic retinopathy was introduced in Denmark in 1988. At present, screening is carried out in four of the 14 counties, thus being available to a minority of diabetic subjects. According to the WHO but also the National Board of Health, all diabetic patients should undergo screening for diabetic retinopathy. Not only does screening pay off in terms of preservation of vision, it is also cost-effective, as confirmed in the present study. The first fully digitized clinic for retinopathic screening in diabetic patients was recently introduced in the County of North Jutland. Preliminary results from this clinic indicate that all procedures can be handled in this fully digitized system. We therefore predict that fully digitized clinics for retinopathic screening will be introduced throughout Denmark in the future. PMID- 12082763 TI - [MAP kinases--molecular transistors in animals and plants]. AB - The survival of multicellular organisms depends on the ability of their cells to communicate with each other and to respond to environmental changes. A goal of modern biology is to uncover the processes by which these cellular signals are transduced. Recent studies have shown that MAP-kinases (MAPKs) are important constituents of such signal transduction pathways. MAPKs function as modules in phosphorelay cascades to activate or repress the activity of downstream target proteins. For example, recent research with knockout mice has shown that mammalian MAPKs are involved in the control of neuronal apoptosis and the activation of immune responses. These mammalian MAPKs exert their control by both promoting and inhibiting specific processes. Surprisingly, plants also use MAPKs to control their immune responses, and plant MAPKs also seem to play dual roles as positive and negative regulators. Such mechanistic similarities provide the basis for fruitful conceptual exchange between molecular research on animals and plants. PMID- 12082764 TI - [Non-biological liver support in hepatic failure]. AB - Liver failure leads to the accumulation of a range of toxins that may be implicated in the development of the multiorgan failure associated with liver failure. A device capable of removing toxins would thus be useful in the treatment of liver failure by stabilising the patient until either the liver begins to regenerate or liver transplantation can take place. A number of non biological devices based on modifications and combinations of haemodialysis, haemofiltration, haemoperfusion, and plasma separation have been investigated and are reviewed in this article. So far, the most promising results have been demonstrated with devices that remove a wide range of toxins, including those bound to albumin. PMID- 12082765 TI - [Sporadic registration of obesity as secondary diagnosis]. AB - INTRODUCTION: The aim was to examine the extent to which obese patients, who are followed up for an obesity-related disease in an outpatient clinic, are correctly registered with the secondary diagnosis of obesity. MATERIAL AND METHODS: We investigated the number of patients at the Endocrine Outpatient Clinic, Rigshospitalet, Copenhagen, who were registered in the patient administrative system with the primary diagnosis of type 2 diabetes, and how many of these were registered with the secondary diagnosis of obesity. RESULTS: Of 233 patients with type 2 diabetes, 79 had a BMI between 25.0 and 29.9 kg/m2 (overweight) and 108 a BMI > 30 kg/m2 (obesity). Thus, 80% of these patients were overweight or obese. Of the 108 patients with a BMI > 30 kg/m2. only 13 (12%) were registered with the secondary diagnosis of obesity, and of 17 severely obese patients with a BMI > 40 kg/m2 only four (24%) were registered with the secondary diagnosis of obesity. DISCUSSION: Obese patients with type 2 diabetes are seldom correctly registered with the secondary diagnosis of obesity. The actual practice of registration probably causes a large underestimation of the impact, and thereby of the magnitude of the economic cost of and the contribution of obesity to total health care costs. Improved registration of obesity as secondary diagnosis in obesity related diseases is needed. PMID- 12082766 TI - [Obesity in Danish hospitals--treatment in the year 2000]. AB - INTRODUCTION: Obesity is of increasing public health concern. The aim of the study was to investigate the treatment of obesity at Danish hospitals in the year 2000, and to compare the results with a similar study from 1987. MATERIALS AND METHODS: A questionnaire was sent to the heads of departments of internal medicine at all Danish hospitals where such departments exist (64). RESULTS: Ninety-one percent of the questionnaires were answered. Fewer hospitals (76%) in Denmark treated obesity in 2000 compared to 1987. About 1400 patients were referred, and the treatment of 500-900 patients was concluded yearly. Many departments reject patients when the primary diagnosis for referral is obesity, often because they lack resources or do not have the capacity. Except for university hospitals, obesity was more often treated on an outpatient basis in 2000 than in 1987. Apart from surgery, the same methods were used to treat obesity at the different types of hospitals. Instructions for treatment, long term outpatient follow-up, quality assurance, evaluation of the treatment, and research in obesity are carried out to some degree at the university hospitals, and only to a small extent at other types of hospitals. DISCUSSION: Although obesity is becoming an increasing threat to public health, treatment at Danish hospitals has decreased over the last ten years, and in the year 2000 only a few per thousand obese patient were offered treatment by a hospital. Increased focus and resources, as well as development and research targeted specifically on the treatment of obesity are needed. PMID- 12082767 TI - [Obesity in Danish hospitals--attitudes in the year 2000]. AB - INTRODUCTION: The aim was to investigate the attitudes towards obesity of those responsible for the treatment at Danish hospitals. MATERIALS AND METHODS: A questionnaire was sent to the heads of departments of internal medicine at all Danish hospitals where such exist (n = 64). RESULTS: Eighty-nine per cent of the questionnaires were answered. Sixteen per cent believed that obesity is a self imposed, life-style disease, and 56% answered "perhaps". 66% believed that obesity is a chronic condition or disease, and 32% thought that obese patients should be followed up life-long. Most believed that diseases would be prevented or improved, if obesity were treated, and 72% believed that it was possible to treat obesity. Thirty-five percent answered "perhaps" to whether it is worth the effort to treat obesity, as weight loss seldom lasts in the long term, and 25% answered "perhaps" to whether loss of weight in the obese incurs health risk. Fifty per cent believed that more resources were needed for the treatment of obesity, and that research in obesity should be given more attention, whereas only a few answered in the negative (6-9%). Thirty-three per cent believed that there was a need for public obesity clinics. Only 15% thought that Danish doctors are sufficiently trained to treat obesity, and only 9% answered "no" to whether better treatment of obesity required altered attitudes to obesity among Danish health care personnel. DISCUSSION: The heads of departments of internal medicine at Danish hospitals had different attitudes to obesity, but most believed that there is a need for altered attitudes, information, and training of Danish doctors and other health care personnel in the treatment of obesity. PMID- 12082768 TI - [Knowledge of contraception and sexual matters over the last 14 years. An investigation among 9th grade pupils in the municipality of Viborg]. AB - INTRODUCTION: The aim of the investigation was to illustrate changes in the knowledge of ninth grade pupils about sexual matters over the last fourteen years. MATERIAL AND METHODS: Ninth grade pupils in schools in the municipality of Viborg answered a questionnaire about these subjects. Identical questionnaires were given to the same age group in 1986 and 1993. RESULTS: The pupils had less knowledge of contraception than earlier. The other questions about sexuality were answered as well as or better than previously. In 1986 and 1993, boys had less knowledge than girls, but this difference was now equalised. We found no connection between knowledge and age at first coitus or sexual habits, and no connection between knowledge and the sources from which the pupils gained their information. School was, as earlier, the most important source of information, but friends came a close second. Books, mother, and doctor or health visitor were often mentioned as important sources of sexual knowledge. As earlier, the pupils were not satisfied with the sex education and as earlier, they wanted more lessons and external teachers. DISCUSSION: A good knowledge of sexual matters does not mean better sexual habits. School is the most important source of knowledge and is even more important, because the young also educate friends, who are another main source of information. According to the pupils, sex education should be improved. PMID- 12082769 TI - [Sexual activity and contraception habits among adolescents over the last 14 years. An investigation among 9th grade pupils in the municipality of Viborg]. AB - INTRODUCTION: The purpose of the investigation was to illustrate changes in sexual activity, the use of contraceptive methods, and age at the first coitus during the last fourteen years. MATERIAL AND METHODS: Twenty-one ninth grade classes in schools in the municipality of Viborg were asked to complete a questionnaire. Altogether 372 questionnaires were distributed and 344 pupils (92%) were entered in the investigation. Neither the teachers nor the pupils knew anything about the investigation before distribution. Identical questionnaires had been completed by the same age group in 1986 and 1993, which enabled us to compare the results and point out differences. RESULTS: We found no changes in age at the first coitus. One third of the pupils had had their first intercourse, 15% before the age of 15. Previously girls were sexually more active than boys, this difference was now equalised. Most (53%) of the experienced pupils had not had coitus in the preceding four weeks, but, compared with 1986, more of the pupils had been sexually active within the preceding week. As found in 1993, the use of condoms had increased, which was still unchanged. There was a continuing decline in the use of the pill at the first intercourse. However, the pupils changed the condoms for the pills as they became more experienced. Ten per cent of the pupils still did not use any contraception at the first intercourse. Occasional use of contraception was more common. In 1986, 75% used contraception at every intercourse, that proportion had fallen to 51% in the year 2000. DISCUSSION: Age at the first intercourse remained unchanged over the 14 years examined, and the change in contraceptional habits seen in 1993 had persisted, except that the proportion using contraception at every intercourse had decreased. The explanation for not using contraception was generally that the intercourse was unplanned. The girls were at a greater risk of unwanted pregnancies and sexually transmitted diseases, as they, more often than the boys, used unsafe methods of contraception. PMID- 12082770 TI - [Physicians' clinical skills after finishing internship]. AB - INTRODUCTION: The aim of this study was to evaluate Danish interns' clinical skills after their internship in order to identify areas where systematic training is needed. This study was a follow-up of a previous study on a group of interns at the beginning of their internship. MATERIAL AND METHODS: A questionnaire listing 88 practical skills was posted to a group of trainees before and after internship. The interns were asked to rate their level of mastery in each skill on a scale 0-10, where 0 = not mastery/not competent and 10 = mastery/competent. RESULTS: In all, 74 out of 100 trainees returned the questionnaire. In 60 of the skills the trainees reported a higher level of mastery after internship and in 28 procedures the level of skill did not improve. Ten of these are procedures used in emergency settings. DISCUSSION: The reliability of self-reported clinical skills can be discussed, but it is a major concern that skills particularly in emergency procedures do not improve. There is a need to define clear objectives, implement structured training programmes, and assess skills in order to accomplish a reliable and sufficient level of competency. PMID- 12082771 TI - [Peliosis hepatis as the cause of liver affection in a patient treated with immunosuppressive agents]. AB - A case of peliosis hepatis in a 64-year-old woman treated with azathioprine because of a lung transplantation is described. The patient was admitted owing to epigastric pain and jaundice. Sixteen months before admission the patient had had a lung transplantation because of pulmonary emphysema. A liver biopsy showed multiple, dilated, blood-filled liver sinusoids consistent with peliosis hepatis. Besides azathioprine, the patient was treated with cyclosporine and prednisolone, but none of these drugs are known to be responsible for the development of peliosis hepatis. This rare condition, which causes liver affection in patients treated with azathioprine, should be kept in mind. PMID- 12082772 TI - [Renal artery stenosis. Can Doppler ultrasound tell us whom are we to treat?]. PMID- 12082773 TI - [A new ethical order related to health research in developing countries]. PMID- 12082774 TI - [Foreign bodies in the eye! Too big a challenge for the general practice?]. PMID- 12082776 TI - [Nicotine replacement and pregnancy]. PMID- 12082775 TI - [Correlation between the prevalence of schizophrenia and the sociodemographic status in different areas of Copenhagen]. PMID- 12082778 TI - [Difficult meeting between science and market forces]. PMID- 12082777 TI - [Vitamins for prevention of abnormalities. The first clinical trial with pregnant diabetic women is about to start]. PMID- 12082779 TI - [Maternal health services are more than medical check ups]. PMID- 12082780 TI - [Hairdressers live dangerously. Daily exposure to chemicals can cause respiratory tract problems]. AB - Hairdressers are regularly exposed to toxic chemicals contained in bleach, hair spray, permanent wave solutions and hair dyes. Adequate ventilation and appropriate working procedures can reduce exposure, but never completely eliminate the risk for respiratory disease. PMID- 12082781 TI - [Normal doses of visible light can cause mutations in skin]. AB - Therapeutic and undesired effects of long-wave (UVA1) (340-400 nm) and visible light were studied. Mutagenicity were investigated after repeated doses encountered in normal situations in the sun. Laser treatment of naevus flammeus was studied with a new 600 nm laser. Also, the effect of photodynamic therapy on cutaneous lymphomas was reported. Both UVA1 and visible light can induce mutations in normal skin after repeated doses. The findings indicate that clothing, not sunscreens, should form the basis of sun protection. In selected cases, the new 600 nm laser may be advantageous in the treatment of naevus flammeus. Localized cutaneous lymphoma may be effectively treated with photodynamic therapy. PMID- 12082782 TI - [The Berzelius symposium on genetic dermatoses: cutaneous gene therapy--potential treatment of severe skin diseases]. PMID- 12082783 TI - [Emotional exhaustion common among women in the public sector]. AB - This cross-sectional investigation aimed at assessing levels of emotional exhaustion among female employees within the Swedish public sector. Other aims were to study the associations between self-rated emotional exhaustion and psychosocial factors at work, as well as findings from medical examinations. Data was collected by means of questionnaires including the Maslach Burnout Inventory, among 183 women working in geriatric care and 143 employees at the National Social Insurance Office. We found high proportions of emotional exhaustion in both samples (geriatric care = 34%; Social Insurance Office = 26%). Participants with high scores for emotional exhaustion reported more job-strain, less social support at work and more somatic, emotional and cognitive complaints than those with low or intermediate scores. Medical examinations performed on 19 participants with low scores and 41 with high scores for emotional exhaustion revealed significantly more findings among participants with high emotional exhaustion, particularly fatigue, sleep disturbances and cognitive impairment. There were no group differences in terms of depression or other findings. This study shows that individuals at risk for stress-related disorders may be identified using simple questionnaires. Early interventions for stress in the workplace may prevent incapacitating conditions among a great proportion of women working in the public sector. PMID- 12082784 TI - [Serotonin syndrome. Several cases of this often overlooked diagnosis]. AB - During the recent decade an increasing number of inquiries concerning cases of overdoses exhibiting typical signs of the serotonin syndrome have been recorded at the Swedish Poisons Information Centre. Four of these cases are presented together with a review of the literature. All patients had overdosed moclobemide and in one case this was the only drug taken. The other patients had ingested moclobemide together with citalopram (2 cases) and clomipramine (1 case). Moreover, other serotoninergic pharmaceuticals as sertraline and sumatriptan were simultaneously ingested in one case and buspirone in another. Three of the cases had hyperthermia, > 40 degrees C and the same number showed pronounced muscle rigidity, coma and mydriasis. Other severe signs and symptoms upon admission included positive Babinski and trismus in two cases each and seizures in one. All patients received mechanical ventilation. Two were treated with dantrolene sodium and one of them was given cyproheptadine as well. One patient received cyproheptadine treatment alone and another prolonged muscle relaxation. Three patients had a typical short clinical course, whereas one patient developed rhabdomyolysis, DIC and arrhythmias. All patients fully recovered. PMID- 12082785 TI - [Lung injury in neonates. Causes, strategies for prevention and long-term consequences]. PMID- 12082786 TI - [Positive effects of estrogens on cardiovascular health]. PMID- 12082788 TI - [Infection protection requires maintenance in a "over-hygienic" surrounding world]. PMID- 12082787 TI - [Can serum estradiol predict the effect of raloxifene on breast cancer risk?]. PMID- 12082789 TI - [Privatization of health services in developing countries strikes the poor]. PMID- 12082790 TI - [Firearms--a growing threat against public health]. PMID- 12082792 TI - [Concerning toys from the drug industry etc]. PMID- 12082791 TI - [Three comments on the LIFE trial]. PMID- 12082793 TI - [Plasma insulin concentration in patients with ketoacidosis?]. PMID- 12082794 TI - [Keep serum cobalamin determination as a sift-test]. PMID- 12082795 TI - [To describe arthrosis as a "degenerative joint disease" is obsolete]. PMID- 12082796 TI - [Health through peace a task for networks in the making]. PMID- 12082797 TI - Blood transfusion practitioners. PMID- 12082798 TI - Ethical theories and dilemmas. AB - Nurses are daily faced with questions of ethics. Most of these questions are minor, everyday issues that individual nurses can resolve on their own, without the need for involvement of their colleagues, and with no, or few, implications for patient care. From time to time, however, issues with major implications for patient care will arise that require the members of the perioperative team to make significant ethical decisions. In this article, Suzanne Hughes discusses the nature of ethics, the ethical questions that nurses might have to address and the consequences arising from decisions. PMID- 12082799 TI - Changing the workforce. PMID- 12082801 TI - A story of recovery. PMID- 12082800 TI - The role of the RNSA in colorectal and general surgery. AB - More nurses are taking on the role of Surgical Assistant. In this article, Jill Biggins reports on her own experience of becoming a Registered Nurse Surgical Assistant in colorectal and general surgery. Jill explains how the role was developed, its tasks and responsibilities, the response of other perioperative practitioners to the role and how further development of the role might be achieved. PMID- 12082802 TI - What is a whistleblower? PMID- 12082803 TI - Empowerment in the workplace. PMID- 12082804 TI - Milk soy protein intolerance: a mother's perspective. PMID- 12082805 TI - Domestic violence victims: a nurses guide to care. PMID- 12082806 TI - The surplus of drug shortages: a prescription for concern. PMID- 12082807 TI - Ethical issues faced by rural physicians. PMID- 12082808 TI - Dual breast cancers followed by endometrial cancer. AB - Two cases in which dual breast cancers occurred at several years interval, followed by endometrial cancer are described. The risk factors for dual breast cancers, the association between breast and endometrial cancer, and the association of tamoxifen therapy with endometrial cancer, and future cancer risk will be briefly discussed. PMID- 12082809 TI - [Cancer in Denmark--diagnosis and prevention]. PMID- 12082810 TI - [Relative survival of Danish cancer patients diagnosed 1981 to 1997 and followed to 2001. A status report]. AB - INTRODUCTION: The last comprehensive estimation of survival for Danish cancer patients was carried out 10 years ago. Based on this estimation and on Nordic and international survival comparisons, a national cancer plan was formulated in order to improve the poor relative survival in Danish cancer patients as compared to other Nordic countries. MATERIAL AND METHODS: We used the national cancer register data validated until the end of 1997 and followed up patients below 90 years of age, known from sources other than death certificates and autopsies to the end of 2000. One-, five-, and 10-year relative survival were computed for age groups 0-44, 45-64, 65-74 and 75-89 from 1981 and onwards in five-year calendar periods, ending with a two-year period, 1996-1997. RESULTS: Altogether 396,248 patients were studied. The five-year relative survival improved substantially over time for all sites together (except the skin) to 39% for men and 51% for women. This improvement was seen for cancer of the colon, rectum, breast, bladder, kidney, cervix and corpus uteri, testis, brain, haematological cancers, and malignant melanoma. No improvement was seen for cancer of the lungs in particular, the oesophagus, stomach, oral cavity, and the pharynx. In general, the prognosis was better for the younger age groups than the older groups. DISCUSSION: Future analysis compared to the present will demonstrate the immediate effect, and later the long-term effect, of the National Cancer Plan. PMID- 12082811 TI - [Cancer genetics. A review of oncological molecular biology seen in relation to the human genome]. AB - Genetic changes are associated with the neoplastic process, when a normal cell is transformed into a cancer cell. Cell growth and differentiation are regulated by complicated interactions between growth promoting and inhibiting signals. Oncogenes are examples of growth promoting genes and tumour suppressor genes (gatekeepers and caretakers) are examples of growth inhibiting genes. Binding of a growth factor to its membrane receptor induces a cascade of intracellular signals resulting in proliferation. Regulation of proliferation is tightly correlated to the regulation of cell cycle, and many of the growth regulating genes also regulate cell cycle, and vice versa. The malignant transformation also includes evasion of programmed cell death (apoptosis), limitless replicative potential (evasion of senescence), sustained angiogenesis, and tissue invasion and metastasis. A rough sketch of the human genome has recently been published, and it brings hope of new potentials in cancer diagnostics, treatment, and choice of treatment, but also in revealing the molecular mechanisms behind cancer predisposition and environmental interactions. It is to be hoped that this will result in possibilities of cancer prophylaxis. PMID- 12082812 TI - [What is epidemiology and what is cancer epidemiology?]. AB - We have the world's oldest cancer registry in Denmark. We also have a cancer society that has invested substantial funds in epidemiology, which have been well used by highly skilled cancer epidemiologists. This is the formula for success. Cancer epidemiology, which is now conducted at the highest international level in Denmark, is a subdiscipline within epidemiology, and most of its activities are related to public health problems. Clinical cancer epidemiology is, however, a field which will grow in the next decades. Cancer epidemiology requires data of good quality and access to data for use in research. New important results will emerge if trajectories of cancer occurrence can be studied in a lifelong perspective. Research involving genetic factors will also open up new avenues for studies, but it will not replace research, whose starting point is the population's social conditions, their lifestyle, and environmental influences. PMID- 12082813 TI - [Prevention of cancer--what do we do in Denmark and what can we achieve?]. AB - We know from our cancer registers and from epidemiological research that we have a major potential for cancer prevention. If the risk factors had been commonly known in the population and the population had been willing to change their risk behaviours, about one third or perhaps half of all new cases could have been avoided today. Cancer epidemiology has pointed to factors of major importance for public health, such as use of tobacco, dietary factors (including alcohol), physical activity, obesity, environmental exposure in leisure time (sun), and in the workplace. All of these can be subject to primary prevention, i.e. behavior to minimise the individual's risk of contracting cancer. Studies on secondary prevention such as screening for cancer have attracted much attention, perhaps at the expense of secondary prevention related to tobacco, alcohol, and diet in the treatment, thereby avoiding side effects and improving the outcome of therapy. Cancer epidemiology has a long tradition in Denmark, whereas translation of the epidemiological findings into prevention is lagging behind. In future, a close collaboration is needed between health professionals, communication and marketing experts, as well as specialists in education and psychology. Only by joining forces with different disciplines will it be possible to target primary and secondary prevention to ease the burden of disease in the population. Evidently, implementation of preventive measures, will benefit from political will and support. PMID- 12082814 TI - [Health care costs of cancer patients. Resource utilization by cancer patients]. AB - INTRODUCTION: The aim of the study was to estimate the health care costs of cancer patients for each separate year from the time of diagnosis. MATERIALS AND METHODS: The data consist of a 20 per cent sample of the Danish population followed up from 1993 to 1997. Civil registration numbers were used to combine various computerised registers to describe the morbidity, mortality, and health care costs of each individual person in the sample. The calculation of the health care costs for the course of events for cancer patients was carried out as a cross-sectional study. RESULTS: The first year since diagnosis of cancer was by far the most expensive. A cancer patient incurred, on average, costs of DKK 90,000 for the health care sector. In the second year the costs decreased to about one third, and after that they stabilised to between DKK 10,000 and 20,000. DISCUSSION: Our study differs in the fact that health care costs are calculated for cancer patients as opposed to cancer treatment. PMID- 12082815 TI - [The cancer plan. Political will--professional formation]. PMID- 12082816 TI - [Physicians' training when it comes to cancer treatment]. PMID- 12082817 TI - [Screening for cancer. International knowledge and Danish practice]. AB - Screening makes it possible to detect cancer before the disease gives rise to symptoms. A more effective treatment could thus be offered, and patients would then have a better prognosis. If screening works, mortality from a given cancer disease should decline in the screened population. At present three screening tests meet this requirement: 1) Pap smears for cervical dysplasia, with screening started at the latest at the age of 30 and not before the age of 20; 2) Mammography screening for breast cancer in women aged 50-69; and 3) faecal occult blood testing for colorectal cancer in men and women aged 50-74. But screening means the testing of healthy persons for cancer, and it therefore has a number of negative side effects, such as false positive and false negative tests. Whether or not screening is preferable in a given situation therefore depends on how the advantages are weighted against the disadvantages. PMID- 12082818 TI - [Early diagnosis of cancer]. AB - Early diagnosis of cancer makes a positive difference in morbidity and mortality. It requires patients to consult their GP about the symptoms and the physician's alertness to cancer as a possible diagnosis. Patients do not exhibit stable patterns of reaction to the warning signs. These signs are not specific for cancer and are often found in patients without cancer. Physicians, and especially GPs, are left with the challenge of differentiating between the few cases of cancer and the many cases of non-malignant disease, but showing identical symptoms. Positive predictive values of symptoms and even severe ones such as rectal bleeding, persistent cough, and loss of weight, do not exceed 10%. Early diagnosis of cancer is considered achievable by increased awareness of the population and physicians through information and education. PMID- 12082819 TI - [Diagnostic imaging of cancer]. PMID- 12082820 TI - [A re-evaluation of Eurocare 2 cancer study. Ranking of countries by survival can be misleading]. PMID- 12082821 TI - [STI571 (Glivec)--a new drug for the treatment of chronic myeloid leukemia]. AB - Chronic myeloid leukaemia (CML) is characterised by the occurrence of the Philadelphia (Ph) chromosome (9/22 translocation) and the formation of a fusion protein--the BCR-ABL transcript with constitutive activation of the BCR-ABL tyrosine kinase and consequent changes in the intracellular signal transduction, which is responsible for the deregulated myeloid cell proliferation. STI571 (signal transduction inhibition number 571) is a potent and selective inhibitor of the BCR-ABL tyrosine kinase. In the chronic phase of the disease, normal peripheral blood values are achieved within the first month of treatment in the large majority of patients and in many patients also a cytogenic response within the following months. The results in the advanced phase are far less favourable, which is explained by the development of resistance owing to reactivation of the BCR-ABL signal transduction. Side effects are primarily nausea, vomiting, various rashes, oedema, most often in the periorbital region, and musculoskeletal symptoms, including muscle cramps. Perspectives for treatment with STI571 are described, as are combinations with alpha-interferon and other cytostatics with a synergistic profile. PMID- 12082823 TI - [Physical activity, overweight and cancer]. PMID- 12082824 TI - [Diet and cancer. Contradictory results from experimental and epidemiologic research?]. PMID- 12082822 TI - [Follow-up after surgery for breast cancer]. AB - Routine follow-up of patients operated for primary breast cancer is a very expensive service, and it is necessary to clarify what purpose it serves. There is no evidence that routine follow-up of mastectomised patients influences morbidity, mortality, or quality of life. Only patients entering clinical trials or quality programmes should thus be followed-up. However, there is agreement in the literature that these patients should be offered follow-up in order to diagnose loco regional recurrences. Lumpectomised patients may be cured, if recurrence in the ipsilateral breast is detected at an early stage. Therefore, these patients should be followed up at regular visits. It is recommended that patients should be offered clinical examination every six months for the first five years, and once a year thereafter until ten years have elapsed since the primary treatment. Except for mammography of residual mamma at intervals of 12-18 months, there is no indication for any further paraclinical investigations. PMID- 12082825 TI - [Evidence-based knowledge]. PMID- 12082826 TI - [Can we rely on the results of urine microscopy and culture when tests are performed in general practice? 1]. PMID- 12082827 TI - [Can we rely on the results of urine microscopy and culture when tests are performed in general practice? 2]. PMID- 12082828 TI - [Dangerous liaisons]. PMID- 12082829 TI - [Protect health personnel in war zones!]. PMID- 12082830 TI - [Hypothyroidism]. PMID- 12082831 TI - [Stability of body mass index from adolescence to adulthood]. AB - BACKGROUND: The purpose of this study was to track the development of body mass index from adolescence into adulthood and to study the association between parental and offspring's body mass index. MATERIAL AND METHODS: From 1979, 506 children participated in a twenty-years follow-up study. Mean age in 1979 was 13 years. Participants' weight and height were measured in 1979, 1981 and 1991, and self-reported in 1999. Weight and height of parents were reported in 1979 and in 1981. The cohort in this paper was followed up until 1999. RESULTS: In 1999, 26% of women and 53% of men had body mass index (BMI: weight in kg/height in meters2) > or = 25, and 7% of women and 11% of men had BMI > or = 30. Participants in the highest quartile of BMI in 1981 had significantly higher BMI in 1999 compared to those in the lowest 1981 quartile (mean and 95% confidence interval was 27.7 (27.0-28.3) kg/m2 versus 22.0 (21.3-22.7) kg/m2). The highest 1981 quartile also had a higher proportion of overweight and obese participants in 1999 than those in the lowest quartile; 64.8% overweight and 27% obese versus 18.8% overweight and 0.9% obese. Participants whose both parents were in the normal weight range in 1981 had lower mean body mass index in 1999 than those with one parent overweight, and a smaller proportion of them were overweight than among participants with both parents overweight. INTERPRETATION: The probability of overweight in adulthood is predicted by both one's own relative weight during adolescence and by parental relative weight 18 years earlier. PMID- 12082832 TI - [Postoperative urinary retention]. AB - BACKGROUND: Retention of urine is a common postoperative problem. MATERIALS AND METHODS: We present two patients with postoperative urinary retention. Current opinions on monitoring and treatment are reviewed. RESULTS: One male postoperative patient was transferred from the postanaesthesia care unit after orthopaedic surgery under spinal anaesthesia. Shortly thereafter he collapsed. Hypotension and bradycardia were treated with intravenous ephedrine and atropine. Urinary retention was suspected and the bladder catheterised for 1,000 ml urine. A follow-up examination revealed no sequelae. A female patient was operated for haemorrhoids under epidural anaesthesia. The second postoperative day she was catheterised for 1,500 ml of urine. Two years later she had an ovarian cyst removed under general anaesthesia. Postoperative urinary retention developed with bladder overdistension (volume > 1,000 ml). A follow-up examination showed rest urine of 25 to 75 ml, and a chronic bladder dysfunction. The incidence of postoperative urinary retention varies depending upon type of surgery, gender, age, and preoperative history of urinary tract dysfunction. Method of anaesthesia or postoperative pain treatment is not correlated to incidence of urinary retention, but excessive intravenous volume load during the operation is. INTERPRETATION: Postoperative urinary retention should be monitored in all patients after surgery. All patients with history of difficulties with micturition need close supervision to avoid urinary bladder overdistention. PMID- 12082833 TI - [Laparoscopic treatment of splenic cysts]. AB - Splenic cysts are uncommon. The symptoms are usually vague, but complications such as haemorrhage, rupture and infection may occur. The majority of splenic cysts are seen in children and young adults. Surgical treatment is recommended and splenic preservation is advantageous. We present two patients with splenic cysts successfully treated with laparoscopic resection of the cyst wall. PMID- 12082834 TI - [Occult spinal dysraphism]. AB - BACKGROUND: Spinal dysraphism is a group of congenital anomalies of the spine characterized by a midline defect affecting the nervous tissue and its bony and soft tissue coverings. Closed spinal dysraphism, spina bifida occulta, refers to skin-covered lesions. However, there are a number of cutaneous stigmata or other physical signs that serve as indicators of an underlying dysraphic malformation of the lower spinal canal. MATERIAL AND METHODS: Based upon relevant literature, we present a review of occult spina bifida. RESULTS AND INTERPRETATION: The cutaneous stigmata that may indicate an underlying dysraphism are particularly hairy patches, subcutaneous lipomas, capillary hemangiomas, dorsal dermal sinuses and sacral cutaneous pits. The clinical examination of a child with lumbosacral cutaneous stigmata should focus on neurological signs and symptoms, urological disturbances and orthopaedic deformities in the lower limbs. Spinal MRI is the imaging modality of choice in children presenting with features consistent with occult spinal dysraphism. Children with occult spinal dysraphism should be referred to a neurosurgical unit. PMID- 12082835 TI - [Type 2 diabetes and nephropathy--new studies, new treatment strategies?]. AB - BACKGROUND: In industrialised countries the incidence of type 2 diabetes associated end stage renal disease has doubled over the last ten years. It is important to lower the blood pressure to inhibit progression of renal failure and to prevent micro- and macrovascular disease in these patients. There is an ongoing discussion on what should be the drug of choice. MATERIAL AND METHODS: We discuss the results from three landmark studies, recently published, on the use of angiotensin II antagonists in patients with type 2 diabetics and nephropathy. RESULTS AND INTERPRETATION: All three studies found a renoprotective effect of angiotensin II antagonists that could not be explained by the effect on the blood pressure alone. Blockade of the renin angiotensin system with angiotensin II antagonist should be the basis of treatment in type 2 diabetic nephropathy. PMID- 12082836 TI - [Acupuncture--a complementary treatment in general practice]. AB - BACKGROUND: Acupuncture is the complementary treatment most commonly used by general practitioners. This study describes the use of acupuncture among Norwegian general practitioners trained in acupuncture. MATERIAL AND METHODS: By telephone or mail, a questionnaire was presented to 212 general practitioners who had completed training in complementary acupuncture. They were asked to describe the use and effect of acupuncture and their attitude towards acupuncture. RESULTS: Of the 111 physicians who replied (53%), 67 (60%) used acupuncture in their clinical practice. 78% had acupuncture courses of less than four weeks' duration only. 52% stated that acupuncture was the preferred treatment in more than 5% of their patients. About 70-80% used acupuncture as a supplement to conventional treatment. Acupuncture was commonly used in patients with musculoskeletal pain (93%), migraine (66%), and headache (63%), less often in patients with nausea, allergy, anxiety, sleep disturbances and gastrointestinal disorders. Improvement was reported in approximately three out of four patients. Lack of time was regarded as the major limitation to the use of acupuncture. INTERPRETATION: Many general practitioners trained in complementary acupuncture use acupuncture as an integrated part of their clinical practice. PMID- 12082837 TI - [Muller-Harbitz disease--familial hypercholesterolemia]. AB - The first patients with xanthomatosis and cardiovascular disease were described in end of the 18th century. From 1925 to 1938, the Norwegian pathologist Francis Harbitz (1867-1950) published several reports on sudden death and xanthomatosis. Harbitz called attention to certain peculiarities of the xanthomatosis. Microscopically he found that the so-called foam cells are more marked and more characteristic than in senile arteriosclerosis. Carl Muller's (1886-1983) attention was directed to this form of cardiovascular disease by the publications of Harbitz. Based on his own studies of 17 families in Oslo over a short period from 1936, he published his final report in 1939. He described hereditary heart disease due to xanthomatosis and hypercholesterolaemia to be fairly common. It was demonstrated to be a dominant trait in the families. Hypercholesterolaemia was present, most marked in connection with xanthoma tuberosus, but there was no definitive relationship between hypercholesterolaemia and xanthomatous deposits in the skin. The occurrence of heart disease in families should direct attention to this disorder. Carl Muller postulated that causal and prophylactic treatment might prove to be of value, but more than 50 years passed before this was possible in the disorder that is now called familial hypercholesterolaemia. PMID- 12082838 TI - [Treatment of chronic hepatitis C]. PMID- 12082839 TI - [Hypothyroidism--a manifold clinical view]. AB - Symptoms of hypothyroidism are often unspecific and numerous. As a doctor you will not get the diagnosis if you do not think of the disease and take the tests necessary to verify it. Doctors have been criticised for not being able to make the correct diagnosis and give proper treatment to these patients. As a consequence, too many patients are treated. Here we describe four patients who illustrate how conventional radiotherapy after cancer mammae surgery gave rise to hypothyroidism, how fine needle aspiration biopsy of the thyroid gland can help in the diagnosis of autoimmune thyroiditis, and a patient without hypothyroidism who was treated with thyroxine and triiodothyronine. PMID- 12082840 TI - [Treatment of hypothyroidism]. AB - BACKGROUND: Hypothyroidism is a common disease with a prevalence of 5-6% in women and 1-2% in men. Primary hypothyroidism is the most common form (> 98%) autoimmune thyroiditis and thyroid destruction caused by radio-iodine treatment or thyroidectomy are the most common causes. MATERIALS AND METHODS: On the basis of relevant literature, an overview of diagnostic criteria and guidelines for treatment is presented. RESULTS AND INTERPRETATION: An elevated TSH and a low free thyroxine level are diagnostic for primary hypothyroidism. The presence of anti-TPO antibodies indicates an autoimmune thyroiditis as the cause. In patients with non-thyroidal illness, a range of test abnormalities may be observed, making the interpretation of the test results difficult. Thyroxine is the preferred treatment of hypothyroidism. The therapeutic goal is to achieve a TSH level between 0.5-1.5 mIU/l. Most patients will then have thyroxine values in the upper 1/3 of the reference range, some will have elevated thyroxine values. If necessary, the thyroxine dose can be increased to give a TSH level between 0.2 and 1.0 mIU/l. In patients not satisfactorily treated with thyroxine alone, treatment with thyroxine combined with triiodothyronine may be tried. A diagnosis made on clinical grounds must always be verified biochemically before thyroxine treatment is started. The present literature does not support the treatment of patients complaining of "symptoms of hypothyroidism" such as tiredness, lethargy, weight gain and intolerance to cold, provided that their thyroid function tests are normal. PMID- 12082841 TI - ["Subclinical" thyroid disease]. AB - BACKGROUND: It is generally easy to diagnose and treat patients with thyroid diseases, though there has been considerable uncertainty over whether or not to treat patients with subclinical disease. MATERIAL AND METHODS: An overview of clinical symptoms and complications related to subclinical thyroid disease is presented on the basis of relevant literature. RESULTS: Patients with subclinical hypothyreosis (elevated TSH level and normal free T4) may benefit from treatment with thyroxine in relation to well-being, serum lipid levels, and possibly also blood pressure. Patients with subclinical hyperthyreosis (suppressed TSH level and normal free T4) have an increased risk of developing atrial fibrillation and possibly also dementia. INTERPRETATION: Subclinical thyroid disease is usually progressive. If a transient condition is ruled out, proper treatment should not be delayed. PMID- 12082842 TI - [Literature search--life or death?]. PMID- 12082843 TI - [Anticoagulant therapy of atrial fibrillation]. PMID- 12082844 TI - [Autopsy--a valuable tool]. PMID- 12082845 TI - [Uncontrolled use of pills?]. PMID- 12082846 TI - [Oncologic treatment]. PMID- 12082847 TI - [Patient support focusing on life]. PMID- 12082848 TI - [Palliation of pain from bone metastases with radioactive isotopes]. AB - We summarise the physical and pharmacokinetic properties of the radionuclides used. The effect on pain from bone metastases caused by prostate cancer, breast cancer, and lung cancer is well documented. A review of the literature does not substantiate a clinically significant difference in the palliative effect produced by any of the radionuclides used. The effect achieved with nuclides is the same as that seen after external radiation. Radionuclides seem to reduce the number of new painful sites from bone metastases, and, as such, use of nuclides is expected to reduce the need for repeated external palliative radiation. PMID- 12082850 TI - [Brain tumors. Mainly glial tumors in adults]. PMID- 12082849 TI - [Immunotherapy of cancer]. AB - Significant advances in the understanding of the cells and molecules involved in tumour/host interactions in humans have been made over the last decades. Thus, through studies of the interactions between cells of the immune system and cancer cells, several tumour antigens and their epitopes recognised by human leucocyte antigen class I-restricted cytotoxic T lymphocytes have been identified. Likewise, specialised antigen presenting cells have been characterised and methods developed that allow the use of such cells in clinical immunology. In the clinic, systemic administration of immune stimulatory cytokines, in particular IL 2, has strengthened the notion that the immune system is capable of mediating rejection of cancer. More recently, more sophisticated methods of inducing anti cancer immune responses have been pursued, for instance the use of antigen presenting cells in combination with various sources of antigen and cytokines for therapeutic vaccinations against cancer. The present paper summarizes current knowledge in the field of tumour immunology, and reviews various approaches undertaken with the aim of inducing strong anti-tumour responses. To this end, we review data from clinical trials, and discuss the potential of these treatment modalities as an adjunct to conventional modes of cancer treatment. Further, novel methodologies for monitoring and analysing cancer patients over the course of immunotherapeutic treatments are described, and future directions put forward. PMID- 12082851 TI - [Head and neck cancer]. PMID- 12082852 TI - [Breast cancer]. PMID- 12082853 TI - [Lung cancer]. PMID- 12082854 TI - [Malignant skin melanoma]. PMID- 12082855 TI - [Sarcomas]. PMID- 12082856 TI - [Pancreatic cancer]. PMID- 12082857 TI - [Colonic cancer]. PMID- 12082858 TI - [Rectal cancer]. PMID- 12082859 TI - [Surgical treatment of liver metastasis]. PMID- 12082860 TI - [Urinary bladder cancer]. PMID- 12082861 TI - [Ovarian cancer]. PMID- 12082862 TI - [Cervix cancer]. PMID- 12082863 TI - [Testicular cancer]. PMID- 12082864 TI - [Palliative care of non-resectable stenosed esophageal and cardiac cancer. A retrospective study of 31 patients treated with endoscopic argon "beam" coagulation]. AB - INTRODUCTION: The aim of this study was to describe the argon beam coagulation technique in the palliative treatment of patients with non-resectable carcinoma of the cardia and oesophagus and its requirements, complications, and tolerance. MATERIALS AND METHODS: 31 patients, referred for palliation by ABC in a central hospital over a period of nearly four years (1.1.1998-31.8.2001), were evaluated retrospectively. RESULT: Twenty-two men and nine women, median age 72 years (range 49-91), underwent a total of 163 treatments and a median of five treatments per patient (range 1-18). Re-canalisation enabling passage of the scope was achieved in 89% of treatments, and most of the patients had dysphagia grade = 2. The median range between reinterventions was 25 days (range 1-175). Perforation was seen in three patients and in 1.8% of treatments procedure related mortality was 1.2%. The median hospital stay for each treatment was two days (range 1-27) and the median impatient stay as proportion survival time was 8%. The median survival was 190 days (range 7-612) and the one-year survival 19%. DISCUSSION: ABC is a well tolerated, safe, and effective treatment in patients with non-resectable cancer of the oesophagus or cardia and offers and acceptable complication rate and number of reinterventions compared with laser and stent placement. The technique is easy and inexpensive and requires no further restrictions than conventional monopolar electrocoagulation does. PMID- 12082865 TI - [Three cases of thrombosis of the internal jugular vein]. AB - We describe three cases of ultrasonographically diagnosed thrombosis of the internal jugular vein encountered in an oncological department. Vague symptoms often delay the diagnosis, but it should be considered when patients present with subacute unilateral oedema of the neck, root of neck, or periclavicular region. At present, treatment is heparin initially and coumarins for a minimum of three months. PMID- 12082866 TI - [Cancer in siblings of children with cancer]. AB - INTRODUCTION: In some rare inherited disorders, such as Li-Fraumeni syndrome, relatives of children with cancer are at increased risk of cancer. We aimed to assess relations between childhood cancer and sibling risk, and evaluate the influence of recessive conditions in cancer causation. MATERIAL AND METHODS: We did a population-based cohort study in the Nordic countries of 42,277 siblings of 25,605 children with cancer. Children with cancer were identified from records in the five Nordic cancer registries, and their siblings from nationwide population registries. Cancers in siblings were documented through record linkage with cancer registries and compared with national incidence rates. We also assessed cancer incidence in parents to identify familial cancer syndromes. RESULTS: 284.2 cancers were expected in siblings, whereas 353 were diagnosed (standardised incidence ratio 1.24 95% CI, 1.12-1.38). Risk ratios for siblings were highest in the first decade of life (2.59; 1.89-3.46). We excluded 56 families with genetic syndromes linked to cancer, which reduced this ratio from 1.7 to 1.0 (0.7-1.3) for siblings younger than 20 years and from 1.3 to 1.0 (0.8-1.3) for those aged 20-29 years. We found no new patterns of familial cancer that indicated inherited susceptibility, or evidence that recessive conditions might contribute to cancers not explained by syndromes. 40% of cancers in siblings that occurred before age 20 years could be attributed to known genetic factors, whereas 60% remained unexplained. DISCUSSION: Apart from rare cancer syndromes, paediatric cancer is not an indicator of increased risk in siblings. PMID- 12082867 TI - [Evidence-based treatment of back pain 2002]. PMID- 12082868 TI - [WHO and The Lancet: yes to mammographic screening]. PMID- 12082869 TI - [Screening for hereditary hemochromatosis]. PMID- 12082870 TI - [Otitis media and antibiotics 1]. PMID- 12082871 TI - [Otitis media and antibiotics 2]. PMID- 12082872 TI - Oklahoma nurses association plays key role in passing historic ban on smoking. PMID- 12082873 TI - Addressing the nursing shortage: an eclectic, collaborative approach. PMID- 12082874 TI - Tulsa sexual assault program cited for award by Harvard University. PMID- 12082875 TI - A slice of rural nursing in Oklahoma. PMID- 12082876 TI - Practicing with competence. PMID- 12082877 TI - Research to address treatment of panic disorder. PMID- 12082878 TI - Bariatric beriberi. PMID- 12082879 TI - Jejunoileal bypass in the treatment of morbid obesity: a 25-year follow-up study of 36 patients. AB - BACKGROUND: Induction of intestinal malabsorption by jejunoileal (JI) bypass was a widely performed procedure for morbid obesity in the 1970's. The purpose of this study was to evaluate the long-term results. METHODS: A total of 36 patients underwent JI bypass from November 1971 to September 1976. At operation the median age was 33 years and median BMI 42 kg/m2. Shunt lengths varied between 45 and 60 cm. The present check-up of the 28 patients still alive included clinical examination, biochemical tests, bone density measurement and measurement of fecal fat excretion. RESULTS: 10 patients (28%) had had their shunt reversed. With one exception these patients quickly regained weight, and 5 (50%) of them were dead. 23 patients with an intact JI shunt are alive, but 5 of them have had the shunt shortened due to weight gain. Their median age today is 56 years, and median BMI is 30. None of these patients were known to have coronary heart disease or diabetes mellitus at follow-up. Malabsorption of fat is still present. Blind loop syndrome, flatulence, foul fecal smell and diarrhea are the most troublesome long term sequelae. Vitamin and mineral deficiencies are common. 2 of 21 patients (age 80 and 57 years) have osteoporosis. CONCLUSION: When the optimal shunt length for the individual patient is found, JI bypass maintains a substantially reduced weight for 25 years. Vitamin and mineral deficiencies are common, but no serious clinical deficiency states are seen. PMID- 12082880 TI - Effect of vertical banded gastroplasty on hypertension, diabetes and dyslipidemia. AB - BACKGROUND: Operations for morbid obesity that are effective in inducing weight loss improve blood pressure, glycemic control and dyslipidemia. Our purpose was to study the effectiveness of VBG in improving hypertension, diabetes and dyslipidemias in morbidly obese patients. METHODS: Retrospective analysis of a cohort of 80 morbidly, obese patients was conducted. Characteristics of patients were: mean age 37 years; gender: women 65, men 15; mean initial BMI 49.5 kg/m2; follow-up > 5 years in 52 patients. Blood pressure, cholesterol and triglyceride levels and glycemia were assessed preoperatively and 1, 3, 6, 12, 18 months and every year after VBG. RESULTS: Hypertension resolved in 65.5% of patients (38 of 58), serum cholesterol level became normal in 34.28% of patients (12 of 35), triglyceride level became normal in 77.77% (7 of 9) and diabetes resolved in 55.55% (5 of 9). CONCLUSION: Weight loss induced by VBG improves blood pressure, triglyceride levels and glycemia. The beneficial changes occur early in the postoperative period and last 5 years or more, despite of the trend to regain some weight. Improvements in risk factors were greater in patients with higher preoperative values. PMID- 12082881 TI - Peptide YY secretion in morbidly obese patients before and after vertical banded gastroplasty. AB - BACKGROUND: The Peptide YY (PYY) secretion pattern was assessed in morbidly obese (MO) patients before and after vertical banded gastroplasty (VBG). METHODS: 12 MO patients (10 women, 2 men) age 29-62 years, BMI 50.7 +/- 9.6 kg/m2, treated with a VBG were studied. Before surgery, blood samples were taken in basal conditions of fasting and 10, 15, 20, 30 and 60 min after the ingestion of a semiliquid test meal. This was repeated in the same patients 6 and 12 months after VBG. Blood samples were also taken from 6 healthy non-obese subjects as controls. PYY plasma concentration was measured by radioimmunoassay with I125. RESULTS: There were statistically significant differences between the preoperative PYY concentration in MO patients compared to controls. After a VBG, PYY concentration varied significantly compared to the preoperative levels. There was no significant difference between the PYY concentrations in the MO patients after VBG and the controls. CONCLUSION: PYY concentration is lower in MO patients compared with non obese. After VBG, PYY concentration gradually rises to the control levels. PMID- 12082882 TI - A cluster of polyneuropathy and Wernicke-Korsakoff syndrome in a bariatric unit. AB - BACKGROUND: Wernicke-Korsakoff syndrome and peripheral neuropathy are very uncommon in bariatric surgical practice. The literature indicates that these complications tend to strike patients receiving unbalanced diets or undergoing rapid weight-loss. METHODS: In a retrospective analysis of the initial experience of a bariatric team in the city of Belem, Para, in northern Brazil, 5 cases were diagnosed in the first year, 4 of them following gastric bypass and the last one after therapy with an intragastric balloon. RESULTS: All episodes followed periods of severe vomiting, which certainly interfered with intake of food as well as of routine vitamin supplements, resulting in severe polyneuropathy and other neurologic manifestions, mostly damaging motility of lower limbs. Therapy consisted of pharmacologic doses of vitamin B1 along with restoration of adequate diet and multivitamin prescriptions. Physical therapy was employed to prevent atrophy and accelerate normalization of muscle strength. All patients responded to this program after variable intervals without significant sequelae. CONCLUSIONS: Thiamine-related neurologic derangements were a cause for much concern and prolonged morbidity in this series, but responded to vitamin B1 replenishment. A high degree of clinical suspicion in bariatric patients and urgent therapeutic intervention whenever postoperative vomiting persists for several days, especially during the first 2-3 months after operation, are the safest approach to these uncommon episodes. It is speculated whether peculiarities in the regional diet of this area in Brazil could have influenced the high incidence of the neurologic aberrations. PMID- 12082883 TI - Effect of surgically-induced weight loss on leukocyte indicators of chronic inflammation in morbid obesity. AB - BACKGROUND: Recent evidence suggests that morbid obesity is a chronic inflammatory condition that may be associated with immune dysfunction. To test this hypothesis, we investigated several leukocyte cell surface markers of chronic inflammation and followed their response to surgically-induced weight loss. METHODS: 26 patients having Roux-en-Y gastric bypass (RYGBP) for morbid obesity (BMI > 40) were compared to 10 normal controls (BMI < 25). Relative monocyte and neutrophil frequencies and expression of the activation antigens CD11b (adhesion molecule), CD16 (Fc receptor), and CD62L (L-selectin), were evaluated by flow cytometry preoperatively and at 1, 3, 6 and 12 months after RYGBP. Cases served as their own controls but were also compared to non-obese controls. The results were statistically analyzed using Student's t-test and ANOVA for parametric values and Mann-Whitney along with Kruskal-Wallis ANOVA for nonparametric values. RESULTS: The control group had mean age 37 +/- 7.6 with mean 23 +/- 2.5 and no comorbidities. The mean age of the sample group was 40.36 +/- 13.7 with mean BMI 52 +/- 8.2. The neutrophil and monocyte relative frequencies of CD11b (monocytes and neutrophils), and CD16 (neutrophils only) were comparable to controls at baseline and did not change significantly with weight loss throughout the study period. However, a significant reduction of CD62L (L-selectin) expression was noted in monocytes and neutrophils at baseline (neutrophils 103 vs 240 gmf, p < 0.001) (monocytes 104 vs 246 gmf, P < 0.001) when compared to normal controls. Levels of L-selectin normalized by 6 months in both monocytes and neutrophils, and by 12 months had become abnormally elevated in monocytes (monocytes 391 gmf, P = 0.007); in neutrophils, there was an upward trend that did not reach significance. The expression of the LPS receptor CD14 in the study group was elevated significantly compared to controls at baseline (1129 vs 719 gmf, P = 0.004); this marker appeared to return to normal by 3 months. Monocyte CD14+/CD16+ subset percentage were also elevated significantly at baseline (14.3% vs 5.25%, P < 0.001), declined throughout the time period but was still significant at 1 year (8.8%, P < 0.001). Eosinophil percentages were elevated at baseline (3.3% obese vs 1.8% controls, P = 0.003) and remained so throughout the time period. CONCLUSION: Deficiencies in the immune system of morbidly obese individuals include elevated levels of eosinophils, monocyte CD14, and monocyte CD14+/CD16+ subsets, with depression of monocyte and neutrophil CD62L. These abnormal levels reverse rapidly with surgically-induced weight loss. RYGBP is not only a weight loss operation but also appears to be an immune restorative procedure. PMID- 12082884 TI - Insulin resistance in the severely obese and links with metabolic co-morbidities. AB - BACKGROUND: The association between insulin resistance (IR) and obesity and its causal relationship with type 2 diabetes is well recognized. The possibility of an association, causal or otherwise, with other obesity-related co-morbidities warrants consideration. METHODS: IR was calculated pre-operatively in 80 patients undergoing gastric bypass surgery for severe obesity, using the homeostasis model assessment (HOMA) method, and again in 70 patients on at least one occasion post operatively within 12 months. Correlations with weight parameters and pre existing co-morbidities including diabetes, hypertension, dyslipidemia and hepatic steatosis were made. RESULTS: 78/80 patients had IR pre-operatively which did not correlate with pre-operative weight or BMI. As expected, there were positive correlations between pre-operative IR and abnormal glucose tolerance and diabetes. A positive correlation was also found between IR and hepatic steatosis, but no correlation was noted between IR and hypertension or fasting levels of cholesterol, triglycerides or Chol/HDL cholesterol ratios. Improvement in IR was uniformly seen after gastric bypass, sooner than would be accounted for by weight loss alone. The degree of pre-operative IR was not a predictor of weight loss after gastric bypass in these patients. CONCLUSIONS: While IR is an almost universal accompaniment of severe obesity, it does not correlate with the degree of obesity in this group of patients. A number of important co-morbidities show a clear association with IR, and improvement in these after gastric bypass may well be related to striking and rapid changes in IR. PMID- 12082885 TI - The early effect of the bariatric operations on diabetes. PMID- 12082886 TI - Prophylactic cholecystectomy with gastric bypass operation: incidence of gallbladder disease. AB - BACKGROUND: Morbid obesity is one of the major risk factors for gallbladder disease, and this risk is even greater following rapid weight loss. Because of this, prophylactic cholecystectomy has been offered to our patients undergoing the transected silastic ring vertical Roux-en-Y gastric bypass (TSRVRYGBP). A study was undertaken to determine the incidence of pathologic gallbladders in patients undergoing this prophylactic cholecystectomy. METHOD: The records of all patients who underwent TSRVRYGBP from June 1999 through December 2000 were reviewed. Pathologic findings of the gallbladder were documented as cholelithiasis, cholecystitis, cholesterolosis, polyps or normal. RESULTS: 761 patients underwent the operation. 178 patients (23%) had cholecystectomy before the surgery. 154 (20%) had gallstones documented by ultrasound and had cholecystectomy at the time of the surgery. 324 of the 429 patients with negative preoperative findings by ultrasound had pathologic evidence of gallbladder disease. CONCLUSION: Because of the high incidence of gallbladder disease even with negative preoperative findings in morbidly obese patients and the lack of significant morbidity with cholecystectomy in experienced hands, routine cholecystectomy at the time of the weight loss operation is justified. PMID- 12082887 TI - Laparoscopic biliopancreatic diversion: technique and initial results. AB - BACKGROUND: Biliopancreatic diversion (BPD) is an efficient method for treating morbid obesity. We present the BPD technique by laparoscopy and the 1st year follow-up. METHODS: From July 2000 to April 2001, 40 patients underwent laparoscopic BPD (29 women, 11 men), with average age 39 (17-60). Average BMI was 43.6 kg/m2 (38-65). All operations were completed successfuly by laparoscopy. The technique followed strictly the operation of Scopinaro. 7 of these patients had removal of an adjustable gastric band and were converted to BPD. All underwent cholecystectomy. RESULTS: Mean loss of initial excess weight at 3, 6 and 10 months was approximately 30%, 50% and 90%. Mortality was 2.5% (1 patient) due to massive pulmonary embolism. Average operating time was 210 minutes (130-480), and average hospital stay was 4.3 days (3-21). There were 5 postoperative complications (12.5%): 2 pulmonary emboli, 2 GI bleeding from the staple-line, and 1 fistula. There have been 5 late complications (12.5%): 3 cases of diarrhea, 2 elevated PTH with normal calcium, and hypothyroidism that was difficult to control. CONCLUSION: BPD may be performed by laparoscopy without changes in the original technique or in the early results. PMID- 12082888 TI - Laparoscopic standard biliopancreatic diversion: technique and preliminary results. AB - BACKGROUND: A technique for standard laparoscopic BPD was developed. METHODS: Standard laparoscopic BPD was performed in 26 morbidly obese patients with mean BMI 43. Details of the technique, using 6 trocars, and instrumentation are described. Intestinal limb lengths were measured fully stretched, and the gastric remnant volume was also measured. Both enteroenteral and gastrointestinal anastomoses were fashioned with a side-to-side technique using the endoGIA, the conjoined defect being closed with a manual running seromuscular suture. RESULTS: 6 and 12 month weight loss results were similar to those obtained in open BPD. CONCLUSION: Laparoscopic standard BPD is a feasible alternative to the open operative procedure, the major advantage being the likely near total avoidance of wound hernia. PMID- 12082889 TI - Disruption of the leptin-insulin relationship in obese men 24 hours after laparoscopic adjustable silicone gastric banding. AB - BACKGROUND: The placement of a band to attain a tiny stomach pouch has been reported to produce early satiety in patients undergoing gastric banding. The adipocyte-derived hormone, leptin, has been shown to decrease both food intake and body weight. The aim of the present study was to assess the potential involvement of acute changes in leptin concentrations following laparoscopic adjustable silicone gastric banding (LASGB). METHODS: The study groups comprised obese male patients undergoing bariatric surgery by LASGB and overweight men undergoing laparoscopic Nissen fundoplication (NFd). Blood was drawn before surgery and 24 hours postoperatively for glucose, insulin and leptin measurements. RESULTS: In both experimental groups, a statistically significant decrease was observed in pre- and postsurgery glucose (LASGB 111 +/- 8 vs 99 +/- 6 mg/dl, P < 0.01; NFd 107 +/- 7 vs 98 +/- 5 mg/d, P < 0.01) and insulin concentrations (LASGB 39.8 +/- 11.9 vs 32.9 +/- 10.3 U/l, P < 0.01; NFd 13.2 +/- 3.3 vs 12.2 +/- 2.9 U/l, P < 0.05). However, no significant differences were observed when the percent change from pre-surgery values was analysed between both groups. Following surgery, an increase in leptin concentrations was observed in the LASGB group (23.5 +/- 4.7 vs 37.5 +/- 6.8 micrograms/l, P < 0.001) whereas a small decrease was evident in the NFd patients (12.9 +/- 4.6 vs 8.9 +/- 2.2 micrograms/l, P < 0.01). CONCLUSION: These findings strongly suggest that the short-term increase observed in plasma leptin concentrations following LASGB may play a key role in triggering an early satiety signal due to the modification of the gastrointestinal anatomy and physiology. PMID- 12082890 TI - Uncommon intragastric migration of the Swedish adjustable gastric band. AB - BACKGROUND: The aim of this study was to assess the incidence of intragastric migration of the Swedish adjustable gastric band (SAGB) and to evaluate the safety and effectiveness of gastroscopic band removal. METHODS: Between January 1996 and December 2001, 454 patients (381 women, 73 men) underwent a laparoscopic SAGB operation. All data (age, gender, pre- and postoperative weight, time of weight gain, band filling status, endoscopic diagnosis of migration, total weight reduction) were prospectively collected in a computerized data bank. RESULTS: Out of the 454 SAGB operations, 14 (3.1%) intragastric band migrations were observed. The average preoperative weight was 122.2 kg and the average postoperative minimum weight was 80.4 kg. All 14 patients had unexplained weight gain on an average of 20 months after the operation. The average band filling status was 8.2 ml. In 12 patients, the band was removed endoscopically, avoiding laparotomy. The remaining 2 patients are under endoscopic surveillance. The mean operating time was 120 minutes. No peri- or postoperative complication was observed. CONCLUSION: Intragastric band migration is a rare complication and should be considered if a patient starts to regain weight. Migration does not require immediate therapy and therefore this complication could be safely treated endoscopically. PMID- 12082891 TI - The effects of pneumoperitoneum on respiratory mechanics during bariatric surgery. AB - BACKGROUND: The aim of this study was to investigate the influence of laparoscopic and conventional open surgery on respiratory mechanics, and blood gases, and to determine convenient techniques from the point of view of intraoperative respiratory mechanics, for bariatric surgery. METHOD: 40 morbidly obese patients were divided into 2 groups, patients undergoing laparoscopy Group 1, and patients undergoing conventional open surgery Group 2. Resistance of airway, dynamic compliance, and peak inspiratory pressure were measured. Measurement was performed in 4 periods: a) after anesthesia induction, b) after pneumoperitoneum in the Group 1 and after incision in the Group 2, c) after gastric band placement, d) and 5 min before extubation. Blood gases were recorded concomitantly. RESULTS: There was no significant difference between the 2 groups in values of blood gases and respiratory mechanics. CONCLUSION: In the morbidly obese, laparoscopic and open surgery did not cause a significant difference for respiratory mechanics when compared with each other. PMID- 12082892 TI - Laparoscopic adjustable gastric banding at a U.S. center with up to 3-year follow up. AB - BACKGROUND: We present our results from the "B" trial (the 2nd U.S. FDA-approved clinical trial) with the laparoscopic adjustable gastric band (LAGB) or Lap-Band System, regarding weight loss, complications, and effect on co-morbidities with up to 3-year follow-up. METHODS: Between March 1999 and June 2001, 63 morbidly obese patients underwent LAGB following accepted ASBS/SAGES guidelines and protocol requirements. All bands were placed via the classic high peri-gastric dissection above the lesser sac, corresponding to the equator of the calibration tube balloon. Frequent follow-up by a multidisciplinary program was maintained. RESULTS: All procedures were performed laparoscopically with no conversion to laparotomy. Operative time decreased from a mean of 197 minutes for the first 10 patients to 120 minutes for the last group. Average hospital stay was 1.4 days. Perioperative complications included 1 intraoperative gastric perforation, which was closed and did not prevent band placement, and 5 port problems. Gastric slippage occurred in 9 patients (14.2%), 3 of whom were revised to a gastric bypass and 6 who had the band removed. Three additional bands were removed due to infection (1), band erosion (1) and a cluster of gastric symptoms (1). Percent excess weight loss averaged 27.2 at 6 months (range 1-68), 38.3 at 1 year (range 10-77), 46.6 at 2 years (range 16-89), and 53.6 at 3 years (range 21-94). Before surgery, 46 of 63 patients (73%) suffered from a serious comorbidity. Following LAGB, all categories showed marked improvement. CONCLUSIONS: In this study up to 3 years, LAGB provided a safe and sustainable weight loss. Significant resolution of serious co-morbidities was common. A U.S. bariatric practice achieved results comparable to those in the international literature. The primary requisites to achieve optimal results include careful patient selection, the refined surgical technique, and a comprehensive long-term patient management program. PMID- 12082893 TI - Laparoscopic banding: selection and technique in 830 patients. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) with the Lap-Band has been our first choice operation for morbid obesity since September 1993. Results in terms of complications and weight loss are analyzed. METHODS: 830 consecutive patients (F 77.9%) underwent LAGB. Initial body weight was 127.9 +/- SD 23.9 kg, and body mass index (BMI) was 46.4 +/- 7.2 kg/m2. Mean age was 37.9 (15-65). Steps in LAGB were: 1) establishment of reference points for dissection (equator of the balloon inflated with 25 cc air and left crus); 2) creation of a retrogastric tunnel above the bursa omentalis; 3) creation of "virtual" pouch; 4) embedding the band. RESULTS: Mortality was 0, conversion 2.7%, and follow-up 97%. Major complications requiring reoperation developed in 3.9% (36 patients). Early complications were 1 gastric perforation (requiring band removal) and 1 gastric slippage (requiring repositioning). Late complications included 17 stomach slippages (treated by band repositioning in 12 and band removal in 5), 9 malpositions (all treated by band repositioning), 4 gastric erosions by the band (all treated by band removal), 3 psychological intolerance (requiring band removal), and 1 HIV positive (band removed). A minor complication requiring reoperation in 91 patients (11%) was reservoir leakage. 20% of patients who had % excess weight loss < 30 had lost compliance to dietetic, psychological and surgical advice. BMI declined significantly from the initial 46.4 +/- 7.2 to 37.3 +/- 6.8 at 1 year, 36.4 +/- 6.9 at 2 years, 36.8 +/- 7.0 at 3 years, and 36.4 +/- 7.8 at 5 years. CONCLUSION: LAGB is a relatively safe and effective procedure. PMID- 12082894 TI - Laparoscopic adjustable esophagogastric banding: preliminary results. AB - BACKGROUND: Laparoscopic gastric banding is effective in surgical treatment of morbid obesity, but has had the drawback of specific complications, like slippage and gastric erosion. To prevent such complications, modifications have been used, including high retrogastric positioning above the bursa omentalis, complete anterior fixation by gastro-gastric stitches over the band, and reduction of the pouch volume to < or = 15 ml. These technical variants may induce dysphagia. METHODS: We adopted a different technique, consisting of placement of the band (9.75 cm BioEnterics Lap-Band) around the esophagus just above the cardia, to induce an amplification of the dysphagic mechanism. No fixation stitches were used. RESULTS: From January 1999 to March 2001, 80 consecutive patients (16 males, 64 females, mean age 41 years, average BMI 45) were operated this way. All operations were completed laparoscopically. However, in 1 patient the procedure had to be interrupted for bleeding from a large fatty liver injury by the retractor. Complications included 2 cases of slippage: an early one after 24 h, requiring surgical removal, and a late one after 9 months, treated by laparoscopic repositioning. The third complication, a reactive esophageal stenosis, occurred in a transsexual male on estrogen treatment, that needed replacement with a wider Swedish band. Band adjustment was required in 28 patients, one time in 22 cases and twice in the other 6. Mean BMI decreased from 45 to 38 after 6 months, remaining at 37 after 24 months, while excess weight was reduced by 50% at 24 months. CONCLUSIONS: The technique has a re-educational function, in that patients are induced to chew thoroughly, to introduce small morsels of food and to prolong the mastication time, in order to avoid dysphagia. Laparoscopic adjustable esophagogastric banding gave no problem if well positioned, and promoted new alimentary habits through a dysphagic mechanisms, inducing significant excess weight loss. PMID- 12082895 TI - Current status of non-adjustable gastric banding. AB - BACKGROUND: The purpose of this study is to review the current status of non adjustable gastric banding (NGB) and to determine whether this operation is still acceptable in the management of morbid obesity, especially when compared with the adjustable GB (AGB) in long-term results. MATERIALS AND METHODS: A literature search was conducted of data published on NGB and AGB in Obesity Surgery in the past 12 months or available from other sources, with records of early and late band-related complications, reoperation rate and weight loss in groups reporting > or = 100 patients with minimum 3-year postoperative follow-up. RESULTS: 1,812 NGB and 1,968 AGB patients were included. Mean BMI was 42.4 in NGB vs 44.0 in AGB. No statistical difference occurred in the early complication rate (1.4% in NGB vs 1.6% in AGB). A statistical difference was noted in long-term complication rate, (1.9% in NGB vs 6.7% in AGB), and in reoperation rate (3.4% vs 7.2%). There was no difference in excess weight loss at 48 months following both operations (54.2% vs 53.0%). CONCLUSION: A significant difference in favor of NGB occurred in the long-term reoperation rate. No other differences were identified, other than in band material. NGB is a softer material and therefore, according to computerized images, has greater flexibility in copying gastric peristaltic waves, which may result in less irritation and more physiological behavior by this band. PMID- 12082896 TI - Does reduction in gastric acid secretion in bariatric surgery increase diet induced thermogenesis? AB - BACKGROUND: Patients who have undergone gastrectomy for benign ulcer do not develop obesity. Furthermore, morbidly obese patients who undergo biliopancreatic diversion (BPD), Roux-en-Y gastric bypass (RYGBP) and vertical banded gastroplasty (VBG) plus truncal vagotomy, may lose more weight compared with patients who undergo VBG alone. A common characteristic of the above is the reduction of gastric hydrochloric acid secretion. We investigated whether reduction in gastric acid increases dietary-induced thermogenesis because of maldigestion of foods, and this may account for the greater weight loss in the above situations. MATERIALS AND METHODS: 22 volunteers without symptoms from the upper gastrointestinal tract were studied. Gastric pH was measured and resting energy expenditure (MREE), using indirect calorimetry, was determined before and 8 hours after consumption of a standard meal. Parameters were measured again after 2 months administration of proton pump inhibitors in all volunteers. RESULTS: Although significant reduction of gastric acid secretion occurred (p < 0.01), following administration of proton pump inhibitors, the fasting and postprandial MREE remained unchanged (p > 0.05). CONCLUSIONS: The reduction in gastric acid secretion does not increase the energy requirements for digestion of foods and thus is neither the mechanism responsible for the increased weight loss observed after RYGBP or BPD, nor the explanation for the lean appearance of gastrectomized patients. PMID- 12082897 TI - Late gastrointestinal hemorrhage after gastric bypass. AB - BACKGROUND: Hemorrhage from the excluded gastric segment or duodenum after gastric bypass is an uncommon late complication and poses both diagnostic and therapeutic difficulties. We describe 4 cases of late gastrointestinal (GI) hemorrhage after gastric bypass. METHODS: 4 patients who underwent previous Roux en-Y gastric bypass (RYGBP) presented for management of severe GI hemorrhage. Their history, diagnostic work-up, management, and surgical pathology are reviewed. RESULTS: In all 4 patients, preoperative diagnostic evaluation including nuclear scintigraphy, endoscopy, and angiography failed to localize the source of bleeding. Intraoperative endoscopy of the gastric remnant and subtotal gastrectomy were performed in all 4 patients. The mean time interval between RYGBP operation and gastrectomy was 15.5 years (range 13-17 years). In 3 of 4 patients, the source of bleeding was documented on pathologic examination of the resected gastric remnant and duodenum. At a mean follow-up of 15 months, none of the patients developed recurrent GI hemorrhage. CONCLUSION: GI hemorrhage after RYGBP can be a diagnostic and therapeutic dilemma. Intraoperative endoscopy of the excluded stomach and subtotal gastrectomy should be considered when the source of bleeding is not identified by conventional diagnostic techniques. PMID- 12082898 TI - Laparoscopic Roux-en-Y gastric bypass for severe gastroesophageal reflux after vertical banded gastroplasty. AB - BACKGROUND: Vertical banded gastroplasty (VBG) is sometimes associated with complications such as pouch obstruction, dilatation, and gastroesophageal reflux. This occasionally requires surgical revision, in many cases to a Roux-en-Y gastric bypass (RYGBP). CASE REPORT: A 47-year-old woman with severe obesity developed severe symptoms of stenosis of the pouch outlet and gastroesophageal reflux 15 years after VBG. Laparoscopic conversion to a RYGBP was performed. At 9 month follow-up, she lost an additional 32 kg and had complete resolution of her reflux. CONCLUSION: In this patient, laparoscopic re-operative RYGBP produced additional weight loss, and improved gastroesophageal reflux that occurred many years after having a VBG. Laparoscopic conversion of a VBG to RYGBP is feasible, and may confer the benefits of other minimally invasive abdominal procedures to this high-risk patient group. PMID- 12082899 TI - Laparoscopic gastric banding after heart transplantation. AB - BACKGROUND: Obesity often complicates the postoperative course of heart transplant recipients. Laparoscopic adjustable gastric banding (LAGB) represents a minimal invasive therapeutic possibility for weight reduction in non transplanted patients. CASE REPORT: We report a 55-year-old diabetic, morbidly obese male (weight 138 kg, height 173 m, BMI 46 kg/m2) in whom 6 years after orthotopic heart transplantation, LAGB and laparoscopic cholecystectomy were successfully performed. At follow-up of 28 months, the patient has lost 32 kg. His present weight is 106 kg (BMI 35.4). Diabetes improved, and oral diabetic medication was withdrawn. Cyclosporine dosage has not had to be changed after LAGB. CONCLUSION: In morbidly obese transplanted patients, LAGB should be considered as an effective alternative to dietetic measures to enable weight reduction and to improve comorbidities. In contrast to bariatric malabsorption techniques, like jejunoileal bypass and gastric bypass, cyclosporine pharmacokinetics do not appear to be influenced by gastric banding. PMID- 12082900 TI - End-stage cardiac failure in a morbidly obese patient treated by biliopancreatic diversion and cardiac transplantation. AB - A 57-year-old 132-kg female (BMI 48.6) with end-stage cardiac failure is presented. Because of a progressive cardiomyopathy, the patient required a heart transplant. However, before receiving the transplant, major weight loss was deemed to be mandatory. Supervised dieting had manifestly failed, and thus a highly effective weight-reducing operation, biliopancreatic diversion, was performed. This resulted in a weight loss of 50.4 kg in 9 months. She then underwent successful cardiac transplantation, and progressed well for several months. She then developed severe psychiatric depression, withheld her medications and developed a lethal acute rejection phenomenon. PMID- 12082901 TI - Peptic ulcer after gastric bypass: should a paracrine pathway be invoked? PMID- 12082902 TI - Sibutramine warning: hypertension and cardiac arrhythmias reported. PMID- 12082903 TI - Walking the moral tightrope: respecting and protecting children in health-related research. PMID- 12082904 TI - Mental competence, caregivers, and the process of consent: research involving Alzheimer's patients or others with decreasing mental capacity. PMID- 12082905 TI - Genetic testing. PMID- 12082906 TI - The psychological structure of patient autonomy. PMID- 12082907 TI - Can there be a "duty to die" without a normative theory? PMID- 12082908 TI - The virtue of moral responsibility in healthcare decisionmaking. PMID- 12082909 TI - Relational professional autonomy. PMID- 12082910 TI - CQ sources/bibliography. Autonomy, the delicate balance. PMID- 12082911 TI - The tyranny of judicial formalism: oral directives and clear and convincing evidence standard. PMID- 12082912 TI - Asylum evaluations--the physician's dilemma. PMID- 12082913 TI - When ethics, healthcare, and human rights conflict: mental healthcare for asylum seekers. PMID- 12082914 TI - How to approach an elevated ferritin level? PMID- 12082915 TI - I.v. heparin for acute coronary syndrome? PMID- 12082916 TI - Patient notes: Tick-borne illness. PMID- 12082917 TI - The looming crisis in geriatric care. PMID- 12082918 TI - Poor image quality leaves physicians in the dark. PMID- 12082919 TI - Overview of causes of COPD. New understanding of pathogenesis and mechanisms can guide future therapy. AB - Recent studies have greatly expanded the understanding of pathogenetic mechanisms underlying COPD. Cigarette smoking is the most important cause. Activation of inflammatory mechanisms in the lung leads to tissue damage by a variety of mechanisms. Active proteases and oxidants are believed to play crucial roles. In response to this injury, repair mechanisms are activated. Unfortunately, these repair mechanisms also may be adversely affected by cigarette smoking. Insufficient repair of alveolar structures in response to injury likely leads to emphysema. Repair with scarring likely leads to airflow limitation in chronic bronchitis. Moreover, mediators that drive these processes have systemic effects that contribute to the syndromes characteristic of COPD. Understanding these mechanisms has created novel therapeutic approaches to the management of this disorder. PMID- 12082920 TI - Early detection and management of COPD. What you can do to reduce the impact of this disabling disease. AB - Primary care physicians play a pivotal role in the early identification of patients with COPD. Early diagnosis, along with aggressive intervention, is the only way that the increasing morbidity and mortality of COPD can be reversed. Any patient with a history of smoking should undergo evaluation for the possibility of underlying COPD. At minimum, this requires in-office spirometry. Smoking cessation is the most important measure to prevent further deterioration of lung function. Rigorous application of a stepped-care treatment plan can help physicians identify undiagnosed COPD and improve overall quality of life for patients who have symptomatic COPD. PMID- 12082921 TI - Treatment of COPD exacerbations. Pharmacologic options and modification of risk factors. AB - Exacerbations of COPD are a major source of morbidity and mortality, resulting in tremendous increases in healthcare costs. Spirometric testing of at-risk persons can help identify patients early in the disease course who may benefit from early intervention to slow the disease process. Avoidance of irritants, smoking cessation, and use of pharmacologic agents aimed at decreasing airflow obstruction are strategies for reducing the frequency and severity of exacerbations. Patients should be educated about modifiable risk factors, such as cigarette smoking, environmental exposures, improper inhaler technique, influenza and pneumonia vaccination, pulmonary rehabilitation, and use of supplemental oxygen. Early, aggressive interventions are necessary to improve quality of life, decrease hospital admissions, improve morbidity and mortality, and reduce overall healthcare costs. PMID- 12082922 TI - Control of chronic nasal symptoms. Directing treatment at the underlying cause. AB - In most patients with rhinitis, careful history taking and physical examination elicit the correct diagnosis. A combination of appropriate environmental and pharmacologic measures is usually effective. Treatment directed at the underlying cause relieves symptoms and improves quality of life. For patients who are unresponsive to treatment, referral to an allergist should be considered. More extensive evaluation and diagnostic testing may reveal previously unrecognized causes of rhinitis. PMID- 12082923 TI - Preparing the patient for renal replacement therapy. Teamwork optimizes outcomes. AB - Proper preparation of a patient with CKD for the development of ESRD and the need for RRT is essential to optimize the patient's quality and quantity of life and to help ensure positive economic and societal outcomes. A collaborative team approach involving the primary care physician team, the patient and his or her family and friends, and the nephrology team should result in improved care of the CKD patient and improved outcomes. It is not possible, feasible, or practical to attempt to provide the inclusive care necessary to attain these goals in a system that does not take advantage of the strengths of a team approach. Adopting this concept of care for patients with kidney disease results in a win-win situation for all of the participants--the patients, the physicians, and society. PMID- 12082924 TI - [Decision in principle concerning HLA typing in England. The Brits allow matched babies]. PMID- 12082925 TI - [Unexpected pharmacological effect resulted in discovery of ghrelin. A new hormone of significance for obesity research]. PMID- 12082926 TI - [Increased mortality among infants born at night. A reflection of inadequate perinatal intensive care resources in the night-time?]. PMID- 12082927 TI - [Smallpox--eradicated disease and a potential terrorist weapon]. AB - Smallpox is a disease that followed humanity for thousands of years up until thirty years ago. It was possible to eradicate, since an effective live vaccine from crossreacting vaccinia could be developed. Twenty years passed since vaccinations stopped and very few people are protected against the disease today. Variola today has become an object of discussion due to the possibility that it can be used as a bioweapon. Due to the number of complications that can be expected a general vaccination is probably not possible. Research is ongoing to develop new vaccines. Many countries are improving their capabilities to respond to a renewed threat of a smallpox epidemic. PMID- 12082929 TI - [Timing of birth and early neonatal mortality in Sweden 1973-1995]. PMID- 12082930 TI - [Follow-up of low birth weight infants at the age of 20 years. Verified deviations remain in adulthood]. PMID- 12082928 TI - [Ghrelin--peptide hormone that increases appetite. It opens new possibilities for the research on energy metabolism]. AB - Ghrelin is an endogenous ligand for the growth hormone secretagogue receptor (GHS R) that potently stimulates growth hormone (GH) release. The discovery of ghrelin opens up a new regulatory system for GH secretion. Surprisingly, recent studies in rodents suggest that peripherally or centrally administered ghrelin, independent of GH, decreases fat oxidation and increases food intake and adiposity. In addition, plasma ghrelin levels are lower in obese human subjects. Ghrelin might participate in meal initiation and may signal to the hypothalamus when an increase in metabolic efficiency is needed. PMID- 12082931 TI - [Great differences in health behavior among high school students. A 5-year follow up of annual questionnaires among students in different school programs]. AB - The school health authority in the municipality of Kramfors performs an annual questionnaire among first-year high school students, which concerns their perceptions about health, wellbeing and the use of alcohol and tobacco. This study covers the period 1995-2000. Psychosomatic complaints and depressive feelings were twice as common among girls. A higher rate of girls and boys in preparatory practical programs were regular users of tobacco and alcohol, were not having breakfast regularly and were practising physical training more seldom compared to girls and boys in more theoretical programs. Bullying was more frequent among boys, especially those from practical programs. In consequence, the school health institutions should emphasise support for exposed groups. A local questionnaire can be a useful tool for identifying such groups and for monitoring health behaviour and health experiences among students as a whole. PMID- 12082932 TI - [NSAID demonstrate convincing effect on postoperative pain]. AB - Non-steroidal antiinflammatory drugs (NSAID) are increasingly being included in the management of postoperative pain as part of a multimodal therapeutic approach. However, there is an ongoing debate among clinicians as to the perioperative use of NSAID's as many authors have cautioned against their use in this setting. Concerns about their impact on coagulation and renal function as well as a risk for gastrointestinal bleeding have been raised. However, many of the adverse effects reported have occurred during long term treatment and might not be directly applicable to the perioperative period. Still the clinician must be aware of the potential risks and benefits of these drugs. This review focuses on the pro's and con's of perioperative NSAID usage. We address the background for a multimodal approach to postoperative pain relief as well as potential alternatives to NSAID's and the recently introduced COX2-selective inhibitors. PMID- 12082933 TI - [Ethics in everyday care. The dialogue is physician's most important tool]. AB - The principle of autonomy brings questions about the dialogue between physicians and their patients in focus. The controversial issue is not if, but when and how to inform patients about their diagnosis, prognosis and treatment options. In difficult situations the main problem is to find a reasonable balance between respect for self-determination and the obligation of beneficence, i.e. the professional responsibility based on assessments of the patient's best interest. One important tool for dealing with this balance in everyday practice is the art of communication with the patient. PMID- 12082935 TI - [The EBM course resulted in critical thinking]. PMID- 12082934 TI - [Evidence-based medicine--a new course in medical education]. PMID- 12082936 TI - [The Global Alliance helps children to fight diseases]. PMID- 12082937 TI - [Relations within health care are at risk to be deformed if economic control gets the upper hand]. PMID- 12082939 TI - [Is forensic psychiatry a secluded world in need of external clinical scrutiny?]. PMID- 12082938 TI - [The role of midwives for better care in connection with abortions--experiences from rich and poor countries]. PMID- 12082940 TI - [Researchers' statements concerning their economic liaisons]. PMID- 12082941 TI - [Is it possible that the patient has been exposed to violence?Should we ask--and how?]. PMID- 12082943 TI - [Abuse of women is a public health problem. All female patients over the age of 14 should be part of a routine screening program]. AB - Abuse does not appear to be restricted to a specific sociodemographic group of women, nor to a specific period in their lives. Screening for domestic violence provides an opportunity not only to disclose family abuse but also to take action to improve the safety and health of these women. All routine health care of women should incorporate direct questions about abuse, psychological as well as physical and sexual. Staff should be trained in how to integrate routine enquiries about abuse into their daily practice. PMID- 12082942 TI - [A literature review of adverse drug reaction reports. Adverse drug effects can have genetical background]. PMID- 12082944 TI - [Evaluation of nurse-run asthma clinics in primary care. Higher quality for less money?]. PMID- 12082945 TI - [Diabetes and tobacco--a double health hazard]. AB - Smoking is a serious risk factor for cardiovascular disease, retinopathy, and nephropathy in patients with diabetes. Furthermore, epidemiological studies have shown that heavy smokers run an increased prospective risk of developing type 2 diabetes, probably due both to the fact that smoking is a marker for an unhealthy lifestyle and that smoking via nicotine may deteriorate glucose metabolism by negatively influencing insulin sensitivity. In Sweden, data from the National Diabetes Register (NDR) has shown that the prevalence of smoking in type 2 diabetes patients followed in primary health care is almost as high as in the non diabetic population, at least in middle-aged subjects (about 20%). This alarming situation must be dealt with by using new and effective methods to promote anti smoking. If support by group sessions can be organised for patients with type 2 diabetes who are smokers, in combination with pharmacological approaches (nicotine, bupropion) it is hoped that the rate of smoking cessation can substantially increase. Such projects are currently under-way within the primary health care in southern Sweden. PMID- 12082946 TI - [PROGRESS--a trial on blood pressure lowering after stroke and TIA. ACE inhibitors and diuretics reduce the risk of new stroke]. PMID- 12082948 TI - [Positive experiences with tonsillotomy in children]. PMID- 12082947 TI - [Antifungal agents can change the balance between resistant and susceptible Candida species]. AB - The use of fluconazole against yeast infections eradicates susceptible but promotes naturally resistant candida strains as C glabrata and C krusei. C albicans dominated in blood cultures taken at the Karolinska Hospital during the years 1989-2001. There was no trend towards an increase of Candida non-albicans. Restrictive use of fluconazole may explain why the balance between different Candida species with varying susceptibility to fluconazole has not shifted. PMID- 12082949 TI - [Secondary prophylaxis after cerebral infarction without cardiac source of embolism. Anticoagulants are not better than ASA according to a new extensive trial (WARSS)]. PMID- 12082950 TI - [Reduced incidence of dementia in patients treated with antilipemic agents]. PMID- 12082951 TI - [TRP channels--sensitive for heat and cold, capsaicin and menthol]. PMID- 12082952 TI - [References--an important part of the scientific article]. PMID- 12082953 TI - [Palinopsia--a visual phenomenon in the art of El Greco and Salvador Dali]. PMID- 12082954 TI - [Who does need Cipralex and why?]. PMID- 12082955 TI - [The LIFE study: extensive marketing in Denmark. Suggestions to conclusions based on the study]. PMID- 12082958 TI - [Is it correct to receive a bonus in connection with drug prescription?]. PMID- 12082956 TI - [Remarkable contribution on estrogen in climacteric]. PMID- 12082959 TI - [Is there a risk of revision in connection with the drug reform?]. PMID- 12082960 TI - [Who is eligible for mobility service?]. PMID- 12082961 TI - Abdominal wound sepsis associated with gynaecological surgery at King Edward VIII Hospital, Durban. AB - OBJECTIVE: To establish the prevalence of postoperative wound infection in major gynaecological surgery. SETTING: King Edward VIII Hospital (KEH), a large urban referral hospital for the province of KwaZulu-Natal, serving an underprivileged population. METHOD: A prospective study of postoperative wound sepsis in 270 patients who had had emergency and elective laparotomies. Data for each patient were obtained pre-operatively and during the postoperative period. RESULTS: The prevalence of surgical wound sepsis was 11.9%. The wound sepsis rate in emergency, elective and semi-elective cases was 14.6%, 6.5% and 4% respectively. Twenty-three per cent of the patients were HIV-infected, and this group had a wound sepsis rate of 28.6% compared with 6.8% in HIV-negative cases (P = 0.03). CONCLUSION: Surgical wound sepsis in major gynaecological surgery is common at KEH. HIV infection is associated with a high rate of wound sepsis. PMID- 12082962 TI - Should knee-length replace thigh-length graduated compression stockings in the prevention of deep-vein thrombosis? AB - OBJECTIVE: To compare knee-length with thigh-length graduated compression stockings for correct application and rate of compliance when they are prescribed for the prevention of deep-vein thrombosis in surgical patients. METHODOLOGY: Patients who were prescribed graduated compression stockings were prospectively studied in three surgical units at Groote Schuur Hospital from February to June 1997. Knee-length stockings were prescribed in the colorectal unit, while the thigh-length variety were prescribed in the hepatobiliary and trauma units. Patients were observed for the correct application and size of stockings, and the presence of compression bands. A total of 72 patients were studied. RESULTS: One patient in the knee-length group and 7 patients in the thigh-length group were not wearing their stockings. Twenty-one of 30 patients (70%) in the knee-length group and 15 of 42 (35.7%) in the thigh-length group had correctly applied stockings (P = 0.009). CONCLUSIONS: Knee-length are more likely to be correctly applied than thigh-length stockings. Knee-length should replace thigh-length stockings in general surgical patients. PMID- 12082963 TI - Abdominal wall endometrioma following caesarean section. PMID- 12082964 TI - Asymptomatic hyperparathyroidism caused by a giant parathyroid adenoma. AB - On routine investigation a 57-year-old woman was found to have primary hyperparathyroidism caused by a giant parathyroid gland. The gland was removed successfully and histological examination proved it to be a parathyroid adenoma. PMID- 12082965 TI - Skin-sparing mastectomy--increasing the options of patients with breast cancer. PMID- 12082966 TI - Late complications of aberrant appendicular faecaliths. PMID- 12082967 TI - Renal trauma secondary to stab, blunt and firearm injuries--a 5-year study. AB - OBJECTIVE: To review our experience of renal trauma at King Edward VIII Hospital, Durban. DESIGN: Retrospective study over 5 years. SETTING: Tertiary referral hospital in an urban area. PATIENTS: One hundred and seventy-four patients presenting with renal injuries treated at King Edward VIII Hospital. INTERVENTIONS: One hundred and thirty-six patients underwent laparotomy, while 38 were managed conservatively. RESULTS: There were 174 patients with renal injuries (148 male, median age 26 years). Eighty-seven injuries were from firearms, 43 from stabs and 44 from blunt trauma. Of 136 patients who underwent surgery, 60 were managed conservatively at laparotomy, 26 underwent renorrhaphy, 40 nephrectomy and 10 heminephrectomy. There was a 33% complication rate following surgery. The mortality rate was 20% (24%, 21% and 12% for firearm, stab and blunt injuries, respectively). There was no statistically significant difference in mortality for firearm, stab and blunt injuries. Furthermore, there was no difference in mortality and morbidity between those with associated organ injuries and those with isolated injuries (29% v. 21% and 30% v. 16%, respectively), but the difference in mortality became significant when patients with isolated injuries who did not undergo surgery were included (P = 0.0083). CONCLUSION: Renal injuries either in isolation or in association with other injuries carry a high mortality and morbidity rate. Firearm injury patients have a large number of associated injuries and have the highest complication rate. Operative strategy should be conservative unless the kidney is unsalvageable. PMID- 12082968 TI - Benzodiazepine sedation in paediatric dentistry. PMID- 12082969 TI - WHO expert committee on specifications for pharmaceutical preparations. AB - The World Health Organization was established in 1948 as a specialized agency of the United Nations serving as the directing and coordinating authority for international health matters and public health. One of WHO's constitutional functions is to provide objective and reliable information and advice in the field of human health, a responsibility that it fulfils in part through its extensive programme of publications. The Organization seeks through its publications to support national health strategies and address the most pressing public health concerns of populations around the world. To respond to the needs of Member States at all levels of development, WHO publishes practical manuals, handbooks and training material for specific categories of health workers; internationally applicable guidelines and standards; reviews and analyses of health policies, programmes and research; and state-of-the-art consensus reports that offer technical advice and recommendations for decision-makers. These books are closely tied to the Organization's priority activities, encompassing disease prevention and control, the development of equitable health systems based on primary health care, and health promotion for individuals and communities. Progress towards better health for all also demands the global dissemination and exchange of information that draws on the knowledge and experience of all WHO's Member countries and the collaboration of world leaders in public health and the biomedical sciences. To ensure the widest possible availability of authoritative information and guidance on health matters, WHO secures the broad international distribution of its publications and encourages their translation and adaptation. By helping to promote and protect health and prevent and control disease throughout the world, WHO's books contribute to achieving the Organization's principal objective--the attainment by all people of the highest possible level of health. PMID- 12082970 TI - On the adsorption behaviour of saliva and purified salivary proteins at solid/liquid interfaces. AB - Salivary proteinaceous substances are known to play important roles in the formation of the salivary pellicle. The aim of this study was to investigate some aspects of the interfacial behaviour of selected purified salivary proteins, as well as human saliva secretions, using time-resolved in situ ellipsometry. Hydrophobic methylated silica and hydrophilic pure silica were used as test substrates. Experiments were performed in vitro, preferentially in the low concentration range, with samples of fresh human whole resting saliva, parotid resting saliva and submandibular/sublingual resting saliva. The protein fractions investigated were human MUC5B, PRP-1, PRP-3 and statherin, as well as bovine submaxillary mucin (BSM). The results indicate that the amount of material adsorbed was strongly related to the protein concentration in the range investigated for both pure proteins and secretions. Generally, a larger amount of material was adsorbed onto hydrophobic surfaces than onto hydrophilic ones. However, pure PRP-1 adsorbed in similar amounts to both hydrophilic and hydrophobic surfaces in the concentration range investigated and BSM adsorbed in larger amounts at high concentrations on hydrophilic surfaces. Comparison of the observed adsorption rates for salivary secretions and calculated diffusion rates for individual proteins suggested initial adsorption of low-molecular-weight proteins/peptides. On hydrophilic surfaces the data indicate adsorption of proteins with diffusion rates corresponding to those of statherin, PRP-3 and PRP 1. MUC5B adsorbs at a later stage from both HWS and the individual secretions, which can be explained by a "Vroman effect"-like phenomenon. On hydrophobic surfaces, adsorption rates were found to be faster than those calculated for any of the proteins, and thus smaller proteins/peptides appear to be involved. The similar adsorption behaviour of PRP-1 and parotid saliva (HPS) on hydrophilic surfaces may suggest that long aPRPs account for a substantial portion of the film-forming capacity of HPS. Effects of added electrolyte could be explained by general screening effects and specific Ca2+ binding to serine phosphates in aqueous solutions, but were complex in phosphate buffer. Inter-individual differences in amounts adsorbed from HWS, HPS and HSMSLS, respectively, were not found to be statistically significant. PMID- 12082971 TI - Remarks on blindness in Denmark and on cooperation between the Danish Association of the Blind and the Danish Ophthalmological Society. PMID- 12082972 TI - The Society's archives. AB - The archives and minutes of the Ophthalmological Society have now been donated to the Medical History Museum of the University of Copenhagen. The first two minutes in the archives are hand-written and cover the period 1900-52. The third minute, mainly typewritten, goes up to 1971. The rest consists of papers, photos, albums, correspondence, gifts etc. Also mentioned are venues for the meetings from Kongens Nytorv (Royal New Square) to various hospitals in Copenhagen etc., finances, many celebrations, meetings, demonstrations, discussions relating to papers, Nordic meetings, relations with opticians, engineers (lighting), printing and certificates (driving, seamen). The archives call for further investigation. PMID- 12082973 TI - Objects relating to the Danish Ophthalmological Society at the Medical History Museum. AB - Some of the objects from the Medical History Museum, University of Copenhagen, concerned with the century-old Danish Ophthalmological Society are mentioned, together with some objects concerned with our predecessors. Jannik Bjerrum's campimeter at 1-2 m-distance reveals glaucomatous arcuate scotoma. [figure: see text] Marius Tscherning's many scientific papers and his ophthalmophacometer (also measuring accommodation), abberoscope, adaptoscope, colour vision, punctual meniscen spectacle glasses etc. are housed in the Museum. K.K.K. Lundsgaard's spectacle collection and cataract paper, Gordon Norrie's colour vision lantern and his personal pince-nez are described. Gustav Osterberg's contact lens practice and his pictorial visual chart, Jorn Boberg-Ans' pioneer artificial lens surgery dating from 1958, are all examples of the work of members of the Society. From our predecessors, we refer to the first cataract extraction in Denmark in 1670 (F. Borri on a goose); Georg Heuermann's on humans dating from 1755 and the reintroduction of the operation in 1810 by C.C. Withusen, V. Krenchel's (1844 1885) ophthalmoscope with a movable mirror and his colour vision lantern; A. Stadfeldt's instruments for examination of the refractive index of the human lens, MD thesis dating from 1898. In addition, snow goggles from Greenland and eye-baths are described. PMID- 12082974 TI - Eye health service in Greenland. PMID- 12082975 TI - Eye health service in the Faroe Islands. PMID- 12082976 TI - Ophthalmology in Iceland. PMID- 12082977 TI - Danish ophthalmology from 1950 to 1975. PMID- 12082978 TI - The period 1975-2000 with a view to the future. PMID- 12082979 TI - The history of the Ophthalmological Society of Copenhagen 1900-50. PMID- 12082980 TI - Developing an ethical view on the use of animals in biomedical research. Fourth Walter C. Randall Lecture on Biomedical Ethics. PMID- 12082981 TI - Science outreach--the responsibility of every scientist. PMID- 12082982 TI - Hatch supports therapeutic cloning. PMID- 12082983 TI - Environmental health services in Europe 6. The development of professional associations. PMID- 12082984 TI - The Japanese encephalitis serological group of flaviviruses: a brief introduction to the group. PMID- 12082985 TI - Japanese encephalitis vaccines: current vaccines and future prospects. AB - Vaccination against JE ideally should be practiced in all areas of Asia where the virus is responsible for human disease. The WHO has placed a high priority on the development of a new vaccine for prevention of JE. Some countries in Asia (Japan, South Korea, North Korea, Taiwan, Vietnam, Thailand, and the PRC) manufacture JE vaccines and practice childhood immunization, while other countries suffering endemic or epidemic disease (India, Nepal, Laos, Cambodia, Bangladesh, Myanmar, Malaysia, Indonesia and the Philippines) have no JE vaccine manufacturing or policy for use. With the exception of the PRC, all countries practicing JE vaccination use formalin inactivated mouse brain vaccines, which are relatively expensive and are associated with rare but clinically significant allergic and neurological adverse events. New inactivated JE vaccines manufactured in Vero cells are in advanced preclinical or early clinical development in Japan, South Korea, Taiwan, and the PRC. An empirically derived, live attenuated vaccine (SA14 14-2) is widely used in the PRC. Trials in the PRC have shown SA14-14-2 to be safe and effective when administered in a two-dose regimen, but regulatory concerns over manufacturing and control have restricted international distribution. The genetic basis of attenuation of SA14-14-2 has been partially defined. A new live attenuated vaccine (ChimeriVax-JE) that uses a reliable flavivirus vaccine--yellow fever 17D--as a live vector for the envelope genes of SA14-14-2 virus is in early clinical trials and appears to be well tolerated and immunogenic after a single dose. Vaccinia and avipox vectored vaccines have also been tested clinically, but are no longer being pursued due to restricted effectiveness mediated by anti-vector immunity. Other approaches to JE vaccines- including naked DNA, oral vaccination, and recombinant subunit vaccines--have been reviewed. PMID- 12082986 TI - Japanese encephalitis virus: ecology and epidemiology. PMID- 12082987 TI - Control of Japanese encephalitis in Japan: immunization of humans and animals, and vector control. PMID- 12082988 TI - Host genetic resistance to Japanese encephalitis group viruses. PMID- 12082989 TI - Pathogenesis and clinical features of Japanese encephalitis and West Nile virus infections. PMID- 12082990 TI - The ecology and epidemiology of West Nile virus in Africa, Europe and Asia. PMID- 12082991 TI - The emergence of West Nile virus in North America: ecology, epidemiology, and surveillance. AB - In late summer 1999, the first domestically acquired human cases of WN encephalitis were documented in the USA. Aggressive vector-control and public education efforts by state and local public health officials limited the extent of human involvement. The discovery of virus-infected, overwintering mosquitoes during the winter of 1999-2000, predicted renewed virus activity for the following spring, and prompted early season vector-control activities and disease surveillance efforts in NYC and the surrounding areas. These surveillance efforts were focused on identifying WN virus infections in birds and mosquitoes as predictors of the potential risk of transmission to humans. By the end of the 2000 mosquito-borne disease transmission season, WN virus activity had been documented as far north as the states of Vermont and New Hampshire, and as far south as the state of North Carolina. The ongoing impacts that WN virus will have on wildlife, domestic animal and human populations of the western hemisphere are not yet known. Plans are in place for public health officials and scientists to monitor the further expansion of WN virus with the establishment or enhancement of vector-borne disease surveillance and control programs throughout the eastern seaboard. The valuable lessons learned from the detection and response to the introduction of WN virus into NYC should prove useful if and when subsequent intrusions of new disease agents occur. PMID- 12082992 TI - Potential vectors of West Nile virus in North America. PMID- 12082993 TI - The ecology and epidemiology of Kunjin virus. PMID- 12082994 TI - West Nile virus in livestock and wildlife. AB - WN virus is one of the most ubiquitous arboviruses occurring over a broad geographical range and in a wide diversity of vertebrate host and vector species. The virus appears to be maintained in endemic foci on the African continent and is transported annually to temperate climates to the north in Europe and to the south in South Africa. Reports of clinical disease due to natural WN virus infection in wild or domestic animals were much less common than reports of infection (virus isolation or antibody detection). Until recently, records of morbidity and mortality in wild birds were confined to a small number of cases and infections causing encephalitis, sometimes fatal, in horses were reported infrequently. In the period 1996-2001, there was an increase in outbreaks of illness due to WN virus in animals as well as humans. Within the traditional range of WN virus, encephalitis was reported in horses in Italy in 1998 and in France in 2000. The first report of disease and deaths caused by WN virus infection in domestic birds was reported in Israel in 1997-1999, involving hundreds of young geese. In 1999 WN virus reached North America and caused an outbreak of encephalitis in humans in the New York area at the same time as a number of cases of equine encephalitis and deaths in American crows and a variety of other bird species, both North American natives and exotics. Multi-state surveillance for WN virus has been in place since April 2000 and has resulted in the detection of WN virus in thousands of dead birds from an increasing number of species in North America, and also in several species of mammals. The surveillance system that has developed in North America because of the utility of testing dead birds for the rapid detection of WN virus presence has been a unique integration of public health and wildlife health agencies. It has been suggested that the recent upsurge in clinical WN virus infection in wild and domestic animals as well as in humans may be related to the emergence of one or more new strains of WN virus. Virus isolated in New York in 1999 was found to be identical to that from Israel. It was alarming for WN virus to so easily invade the United States and surprising that it became established so quickly in the temperature climate of New York. Its persistence and rapid expansion in the United States leave a number of unanswered questions. New disease characteristics and patterns have occurred and more are evolving as WN virus further invades the western hemisphere. Additional animal research is needed to answer these questions. Some of the research needs include bird migration as a mechanism of virus dispersal, vector and vertebrate host relationships, virus persistence mechanisms, laboratory diagnosis, viral pathogenesis, risk factor studies, vaccine development, and WN virus impact on wildlife (CDC 2001a). Determination of the primary reservoir host species that are involved in the epidemiology of WN virus and the suitable sentinel species for active surveillance are also important research areas. PMID- 12082995 TI - The role of birds in the ecology of West Nile virus in Europe and Africa. AB - Surveys on wild birds conducted during the last two decades in Europe, notably Poland and the Czech Republic, to determine their infection rate with WN virus have revealed endemic foci of infection. Some species of seropositive birds were nonmigrators while others were hatchlings of migrating species. Persistently infected avian reservoirs are potential sources of viruses for mosquitoes that multiply in the temperate European zone in hot, wet summers. In the past, evidence for geographical circulation of WN viruses was based on antigenic analysis of strains from different countries while more recent epidemiological studies have relied on analysis of nucleotide sequences of the envelope gene. With the reappearance of epidemic WN fever in European countries, interest has been focused once again on the African origin of the causal agent carried by migrating wild birds. In some epidemics, isolates were made from human cases or mosquitoes and only serologic evidence for infection was available from domestic and wild bird populations. In this respect the unusual findings of anti-WN virus antibodies in a population of storks maintained in northern Germany could be interpreted as evidence for local infection. The unique susceptibility of young domestic geese in Israel in 1997-2000 to WN virus and the isolation of similar strains from migrating White storks in Israel and Egypt suggest that the recent isolates are more pathogenic for certain avain species and that migrating birds do play a crucial role in geographical spread of the virus. Knowledge of the routes taken by birds migrating between Africa and Europe will therefore help in selecting sites where attempts to isolate viruses will be most fruitful. The appearance of the disease in western European equine populations (Italy and France) requires that other birds and their migratory routes be investigated once more. It remains to be determined whether the European endemic foci of WN virus are in themselves sources of infection for other birds that migrate across Europe and do not necessarily reach sub-Saharan Africa. If this is the case it will be necessary to define the strategies for detection of virus overwintering in the European temperate climate. PMID- 12082996 TI - Replication and gene function in Kunjin virus. PMID- 12082997 TI - Interaction of West Nile and Kunjin viruses with cellular components during morphogenesis. PMID- 12082998 TI - Phylogeny and molecular epidemiology of West Nile and Kunjin viruses. PMID- 12082999 TI - Evolution of the Japanese encephalitis serocomplex viruses. PMID- 12083000 TI - Japanese encephalitis as an emerging virus: the emergence and spread of Japanese encephalitis virus in Australasia. PMID- 12083001 TI - Molecular epidemiology of Japanese encephalitis virus. PMID- 12083002 TI - Immune responses to Japanese encephalitis virus. PMID- 12083003 TI - Molecular evolution of proteasomes. AB - Proteasomes are large, multisubunit proteases that are found, in one form or another, in all domains of life and play a critical role in intracellular protein degradation. Although they have substantial structural similarity, the proteasomes of bacteria, archaea, and eukaryotes show many differences in architecture and subunit composition. This article discusses possible paths by which proteasomes may have evolved from simple precursors to the highly complicated and diverse complexes observed today. PMID- 12083004 TI - Natural substrates of the proteasome and their recognition by the ubiquitin system. AB - The multitude of natural substrates of the 26S proteasome demonstrates convincingly the diversity and flexibility of the ubiquitin/proteasome system: at the same time, the number of pathways in which ubiquitin-dependent degradation is involved highlights the importance of regulated proteolysis for cellular metabolism. This review has addressed recent advances in our understanding of the principles that govern the recognition and targeting of potential substrates. While the mechanism of ubiquitin activation and conjugation is largely understood, the determination of substrate specificity by ubiquitin protein ligases remains a field of active research. Several conserved degradation signals within substrate proteins have been identified, and it is becoming increasingly clear that these serve as docking sites for specific sets of E3s, which in turn adhere to a number of well-defined strategies for the recognition of these motifs. In particular, RING finger proteins are now emerging as a new and apparently widespread class of ubiquitin ligases. The discovery of more and more E3s will undoubtedly reveal even better the common principles in architecture and mechanisms of this class of enzymes. In contrast to substrate recognition by the ubiquitin conjugation system, the way in which a ubiquitylated protein is delivered to the 26S proteasome is poorly understood. There is no doubt that multiubiquitin chains serve as the principal determinant for recognition by the proteasome, and a number of receptors and candidate targeting factors are known, some of which are associated with the proteasome itself; however, unresolved issues are the significance of the different geometries that alternatively linked multiubiquitin chains can adopt, the role of transport between subcellular compartments, as well as the participation of chaperones in the delivery step. Finally, the analysis of ubiquitin-independent, substrate-specific targeting mechanisms, such as the AZ-dependent degradation of ODC, may provide unexpected answers to questions about protein recognition by the 26S proteasome. PMID- 12083005 TI - The intracellular localization of the proteasome. AB - In summary, localization of proteasomes does appear to be important in the regulation of proteolysis. In yeast, a discrete localization is observed at the nuclear periphery for cells undergoing mitotic growth. This localization is clearly important as degradation by the ubiquitin/proteasome pathway is impaired in mutants that mislocalize proteasomes. In mammalian cells, proteasomes are present throughout the cell. However, the proteasome does appear to be enriched at the MTOC upon aggresome formation. The inhibition of the ubiquitin/proteasome pathway in aggresome-containing cells could provide an explanation for the pathogenicity of a number of neurodegenerative diseases. PMID- 12083006 TI - Proteasome inhibitors: complex tools for a complex enzyme. AB - As the dominant protease dedicated to protein turnover, the proteasome shapes the cellular protein repertoire. Our knowledge of proteasome regulation and activity has improved considerably over the past decade. Novel inhibitors, in particular, have helped to advance our understanding of proteasome biology. They range from small peptide-based structures that can be modified to vary target specificity, to large macromolecular inhibitors that include proteins. While these reagents have played an important role in establishing our current knowledge of the proteasome's catalytic mechanism, many questions remain. Rapid advances in the synthesis and identification of new classes of proteasome inhibitors over the last 10 years serve as a positive indicator that many of these questions will soon be resolved. The future lies in designing compounds that can function as drugs to target processes involved in disease progression. It may only be a short while before the products of such research have safe application in a practical setting. Structural and combinatorial chemistry approaches are powerful techniques that will bring us closer to these goals. PMID- 12083007 TI - The 20S proteasome. AB - In contrast to our detailed knowledge of prokaryotic proteasomes, we have only a limited understanding of the prokaryotic regulators and their functional interaction with the proteasome. Most probably, we will soon learn more about the molecular structure and the mechanism of action of the prokaryotic regulators. Nevertheless, it still remains to be unravelled which signals or/and modifications transform an endogenous prokaryotic protein into a substrate of the proteasomal degradation machinery. PMID- 12083008 TI - Regulating the 26S proteasome. AB - Despite the fact that the composition of proteasomes purified from different species is almost identical, and the basic components of the proteasome are remarkably conserved among all eukaryotes, there are quite a few additional proteins that show up in certain purifications or in certain screens. There is increasing evidence that the proteasome is in fact a dynamic structure forming multiple interactions with transiently associated subunits and cellular factors that are necessary for functions such as cellular localization, presentation of substrates, substrate-specific interactions, or generation of varied products. Harnessing the eukaryotic proteasome to its defined regulatory roles has been achieved by a number of means: (a) increasing the complexity of the proteasome by gene duplication, and differentiation of members within each gene family (namely the CP and RPT subunits); (b) addition of regulatory particles, complexes, and factors that influence both what enters and what exits the proteasome; and (c) signal-dependent alterations in subunit composition (for example, the CP beta to beta i exchange). It is not be surprising that the proteasome plays diverse roles, and that its specific functions can be fine-tuned depending on biological context or need. PMID- 12083010 TI - Immunological functions of the proteasome. AB - Proteasomes are highly abundant cytosolic and nuclear protease complexes that degrade most intracellular proteins in higher eukaryotes and appear to play a major role in the cytosolic steps of MHC class I antigen processing. This review summarizes the knowledge of the role of proteasomes in antigen processing and the impact of proteasomal proteolysis on T cell-mediated immunity. PMID- 12083011 TI - Is your practice wasting away? PMID- 12083009 TI - The 11S regulators of 20S proteasome activity. AB - Although substantial progress has been made in understanding the biochemical properties of 11S regulators since their discovery in 1992, we still only have a rudimentary understanding of their biological role. As discussed above, we have proposed a model in which the alpha/beta complex promotes the production of antigenic peptides by opening the exit port of the 20S proteasome (Whitby et al. 2000). There are other possibilities, however, that are not exclusive of the exit port hypothesis. For example the alpha/beta complex may promote assembly of immunoproteasome as suggested by Preckel et al. 1999, or it may function as a docking module and conduit for the delivery of peptides to the ER lumen (Realini et al. 1994b). There are also unanswered structural and mechanistic questions. Higher resolution data are needed to discern important structural details of the PA26/20S proteasome complex. The models for binding and activation that are suggested from the structural data have to be tested by mutagenesis and biochemical analysis. What is the role of homolog-specific inserts? Will cognate regulator/proteasome complexes show conformational changes that are not apparent in the currently available crystal structures, including perhaps signs of allosteric communication between the regulator and the proteasome active sites? PMID- 12083012 TI - Patient flow and real estate. PMID- 12083013 TI - New Medicare payment opportunities. PMID- 12083014 TI - Call RVUs: one way to make call more equitable. PMID- 12083015 TI - Achieving a more minority-friendly practice. PMID- 12083017 TI - A simple technology to improve office efficiency. PMID- 12083016 TI - Strategies for better patient flow and cycle time. PMID- 12083018 TI - Security software for hand-held computers. PMID- 12083019 TI - Adding a dose of levity to practice. PMID- 12083020 TI - The National Institutes of Health Consensus Development Conference: Adjuvant Therapy for Breast Cancer. Bethesda, Maryland, USA. November 1-3, 2000. Proceedings. PMID- 12083021 TI - [Child abuse and dentists]. AB - A questionnaire on the recognition and reporting of child abuse was sent to a random sample of Dutch dental practitioners (N = 250). The response rate was 54%. Eighty-six percent had never seen a case of child abuse. The percentage of dentists reporting a (suspicious) case of child abuse (6%) was lower than the percentage of dentists detecting a case of suspicious child abuse (14%). The most important reason for this discrepancy seems to be a lack of knowledge and/or skills to properly identify child abuse. More professional education about child abuse is desired. PMID- 12083022 TI - [The convergence angle of crown preparations with clinically satisfactory retention]. AB - A method was developed to measure the convergence angle on dies of complete crown preparations. The 132 dies measured revealed a surprising range in the convergence angles (15.5 to 30.2 degrees). The mandibular molar preparations had the greatest convergence angle. The design of mandibular molar preparations should be the subject of additional research. PMID- 12083023 TI - [Fit of anchors in frame prostheses]. AB - The fit of clasps was assessed in 101 removable partial dentures (RPD), including 54 free-end prostheses without any tooth bound replacements and 47 tooth bound prostheses replacing only premolars and molars. The age and the design of the RPD had a significant influence on the fit: in older and in stress broken free-end RPD's the clasps had a worse fit in terms of standing away from the abutment tooth. PMID- 12083024 TI - [In the preparation lies the result]. PMID- 12083026 TI - Quality practice environments. PMID- 12083025 TI - [Dental caries in school children in The Hague. A comparison of the research results of 1993 with those from 1989]. AB - Starting in 1969, periodic cross-sectional examinations of schoolchildren have been carried out in the city of The Hague. The results of 1993 show that since 1989 no further change in the caries prevalence of the deciduous and permanent dentition has taken place in 5- and 7-year-old children of Dutch origin. Between 1989 and 1993 a marked DMFS reduction amongst Dutch 11-year-olds was found. Compared to 1981 and 1989 a further reduction of dfs and DMFS values for 7-year old children of ethnic minorities (Turkey and Morocco) could be observed in 1993. Between 1989 and 1993 a significant caries decrease was found among 11-year-old children from Turkey and Morocco. PMID- 12083027 TI - Antibiotics for asymptomatic bacteriuria in pregnancy. PMID- 12083028 TI - Results of stapling and conventional hemorrhoidectomy. PMID- 12083029 TI - [On the work of the Federation of medical historians (international) in 2000 2001]. PMID- 12083030 TI - [The Moscow Scientific Society of Medical Historians in 2001]. PMID- 12083032 TI - A clinician's guide to dental audiences. What every dentist should know before teaching a continuing education course. PMID- 12083031 TI - Eliminating iron deficiency disorders. PMID- 12083034 TI - Lawmakers propose increased funding for AIDS prevention. PMID- 12083033 TI - NIAID, Wyeth create AIDS vaccine team. PMID- 12083035 TI - Genomics. Chimps next for sequencing. PMID- 12083036 TI - HIV vaccine best hope for controlling AIDS, says NNIH. PMID- 12083037 TI - AIDS prevention more cost-effective than HAART treatment. PMID- 12083038 TI - Institute of Medicine: lack of insurance can mean early death. PMID- 12083039 TI - WHO launches program to assist alternative medicines. PMID- 12083041 TI - House passes abstinence-only education funding in welfare bill. PMID- 12083040 TI - HIV-positive inmate's ADA rights not violated by work duty. PMID- 12083042 TI - AIDS infection rate falls among Brazil's injection drug users. PMID- 12083043 TI - HIV-positive flight attendant loses discrimination lawsuit. PMID- 12083044 TI - [Perceptions of social actors about the care delivered by the optical laboratory in the primary school health care program (PROASE) in the municipality of Ribeirao Preto]. AB - This study aimed at evaluating the care delivered by the Optical Laboratory of the Program of Primary Care to Schoolchildren (PROASE) according to the perceptions of relatives and teachers of children who had acquired glasses from 1991 to 1995 in the municipality of Ribeirao Preto. The content analysis technique was used for data analysis and, in order to observe the meanings shown in the qualitative material, the thematic analysis technique was utilized. It was concluded that that the social actors showed a positive attitude concerning the importance of the program and clearly expressed that PROASE has given relevant contribution to the health of schoolchildren. PMID- 12083045 TI - [The use of intestinal enema in labor preparation: analysis of advantages and disadvantages]. AB - This study aimed at investigating the effects of intestinal cleansers on fecal contamination during labor and delivery amongst women assisted in a university hospital. The subjects were 90 pregnant women (43 normal births, 27 forceps and 20 caesarean sections). The population was divided in two randomized groups: with and without the use of intestinal cleansers. It was possible to conclude that intestinal cleansers did not provide faster labor and did not reduce fecal contamination. However, the women who had used an intestinal cleanser tended to regard it as positive. PMID- 12083046 TI - [Technical competence in the prevention of HIV/AIDS: validation of a tool]. AB - This study aimed at building and validating an instrument to measure the technical competence of educational actions developed by nurses in the prevention of HIV/AIDS in Basic Health Units (BHU). The phases for constructing and testing the fidelity of the instrument were: elaboration of items related to the technical competence in the prevention of HIV/AIDS; application of the instrument in a sample of 30 male nurses from different hospitals and teaching institutions. The instrument presented a satisfactory level of stability, homogeneity and reproduction. PMID- 12083048 TI - Got sugar? Tips on preventing diabetes. PMID- 12083049 TI - New STD guidelines issued by CDC. PMID- 12083050 TI - It pays to be picky. PMID- 12083051 TI - Facing the realities of fantasy sex. PMID- 12083052 TI - Addicted to cybersex? PMID- 12083053 TI - When leaving may be the right thing to do. PMID- 12083054 TI - Genderizing HIV/AIDS. PMID- 12083055 TI - Pedal power. PMID- 12083056 TI - The new epidemic. PMID- 12083057 TI - That's life. PMID- 12083058 TI - Medical nutrition therapy now required at many clinics. PMID- 12083059 TI - Bed and breakfast, anyone? PMID- 12083060 TI - [Impact of domestic violence on reproductive and neonatal health]. AB - BACKGROUND: Domestic violence can have an important influence on mother and child health. AIM: To assess the consequences of remote and actual, emotional or physical, domestic violence on the reproductive and newborn health in pregnant women. MATERIAL AND METHODS: A longitudinal epidemiological observation from an Urban Primary Health Care Center from Valdivia, Chile, in 1998. Two cohorts were studied: Pregnant women that experienced domestic violence (index group) and pregnant women not exposed to domestic violence (control group). Women were followed during pregnancy and at labor. The newborn was also assessed. RESULTS: The index group had a higher relative risk (RR) for impending abortion (RR 1.44, 95% confidence interval (CI): 1.07-1.93), hypertensive syndrome of pregnancy (RR 1.5, 95% CI: 1.18-1.96), intrahepatic cholestasis (RR 1.5, 95% CI: 1.1-1.94). Women that experienced violence during pregnancy had a higher risk of urinary tract infection (RR 2.88, 95% CI: 1.28-6.43), intrauterine growth retardation (RR 3.7, 95% CI: 1.77-7.93) and intrahepatic cholestasis. Newborns from the index group had lower weight, size and gestational age. CONCLUSIONS: Domestic violence is associated with hypertension during pregnancy and intrauterine growth retardation. The incorporation of bio-psychological evaluation and monitoring systems could attenuate the consequences of domestic violence. PMID- 12083061 TI - Implant failure. PMID- 12083063 TI - Workplace violence. What factors may be putting you at risk? Interview by Cheryl Anderson. PMID- 12083062 TI - Nicotine and bone density around titanium implants: a histometric study in rabbits. AB - This study investigated the influence of nicotine on bone density around titanium implants inserted in rabbits. Thirty-two New Zealand rabbits were included. After anesthesia, the tibiae surfaces were exposed and two screw-shaped, commercially available, pure titanium implants of 7.0 mm in length and 3.75 mm in diameter were placed bilaterally. A total of 128 implants were inserted: 64 blasted with Al2O3 particles (group 1) and 64 with a machined surface finish (group 2). The animals were randomly assigned to one of four treatment subgroups, and daily subcutaneous injections of nicotine were administered. After 42 days, the animals were killed and undecalcified sections were prepared. The bone density was measured in the cortical passage of the implant. Statistical analysis (two-way analysis of variance) revealed no significant difference neither regarding the effect of nicotine nor the effect of surface design on bone density around the implants (P > 0.05). Within the limits of the present study, it can be assumed that daily nicotine administration may not statistically influence bone density around titanium implants. PMID- 12083064 TI - Pregnancy outcome in HbSS-sickle cell disease in Lagos, Nigeria. AB - A retrospective study of pregnancy outcome in 60 HbSS sickle cell disease patients, exclusively managed in accordance with standard management modality, as obtained in the obstetric Unit of the Lagos University Teaching Hospital (LUTH), was carried out over a 3-year period (1995-1997). The study shows an increase in the number HbSS patients seeking antenatal care in the LUTH. Although the patients maintained a stable haematological profile, all through pregnancy and delivery, as reflected by the booking, pre-delivery and post-delivery PCV of 23.2 +/- SD 4.6% and 23.6 +/- SD 4.5% respectively; pregnancy was complicated in 96.6% of cases (n = 58). Sickle cell crises of bone pain (41.4%), acute haemolysis with anaemia (34.4%); malaria (22.4%) and other systemic infections (22.4%) were the major causes of pregnancy complications. Antenatal and postpartum blood transfusion rates were 45.0% and 81.6% respectively. The perinatal and maternal mortality rates were 121 and 67/1000 respectively, while caesarean section rate was 43.2%. It is observed that while more patients with HbSS sickle cell disease are seeking antenatal care in the LUTH, with improved prognosis, pregnancy complications from sickle cell crisis, remain a major problem in the care of these patients. PMID- 12083065 TI - Aneurysms of the vertebrobasilar tree. PMID- 12083066 TI - Analysis of the aspartic acid metabolic pathway using mutant genes. AB - Amino acid metabolism is a fundamental process for plant growth and development. Although a considerable amount of information is available, little is known about the genetic control of enzymatic steps or regulation of several pathways. Much of the information about biochemical pathways has arisen from the use of mutants lacking key enzymes. Although mutants were largely used already in the 60's, by bacterial and fungal geneticists, it took plant research a long time to catch up. The advance in this area was rapid in the 80's, which was followed in the 90's by the development of techniques of plant transformation. In this review we present an overview of the aspartic acid metabolic pathway, the key regulatory enzymes and the mutants and transgenic plants produced for lysine and threonine metabolism. We also discuss and propose a new study of high-lysine mutants. PMID- 12083067 TI - Molecular and biochemical analysis of serine acetyltransferase and cysteine synthase towards sulfur metabolic engineering in plants. AB - Serine acetyltransferase (SATase) and cysteine synthase (O-acetylserine (thiol) lyase) (CSase) are committed in the final step of cysteine biosynthesis. Six cDNA clones encoding SATase have been isolated from several plants, e.g. watermelon, spinach, Chinese chive and Arabidopsis thaliana. Feedback-inhibition pattern and subcellular localization of plant SATases were evaluated. Two types of SATase that differ in their sensitivity to the feedback inhibition by L-cysteine were found in plants. In Arabidopsis, cytosolic SATase was inhibited by L-cysteine at a physiological concentration in an allosteric manner, but the plastidic and mitochondrial forms were not subjected to this feedback regulation. These results suggest that the regulation of cysteine biosynthesis through feedback inhibition may differ depending on the subcellular compartment. The allosteric domain responsible for L-cysteine inhibition was characterized, using several SATase mutants. The single change of amino acid residue, glycine-277 to cysteine, in the C-terminal region of watermelon SATase caused a significant decrease of the feedback-inhibition sensitivity of watermelon SATase. We made the transgenic Arabidopsis overexpressing point-mutated watermelon SATase gene whose product was not inhibited by L-cysteine. The contents of OAS, cysteine, and glutathione in transgenic Arabidopsis were significantly increased as compared to the wild-type Arabidopsis. Transgenic tobacco (Nicotiana tabacum) (F1) plants with enhanced CSase activities both in the cytosol and in the chloroplasts were generated by cross-fertilization of two transgenic tobacco expressing either cytosolic CSase or chloroplastic CSase. Upon fumigation with 0.1 microLL(-1) sulfur dioxide, both the cysteine and glutathione contents in leaves of F1 plants were increased significantly, but not in leaves of non-transformed control plants. These results indicated that both SATase and CSase play important roles in cysteine biosynthesis and its regulation in plants. PMID- 12083068 TI - Molecular and biochemical analysis of the enzymes of cysteine biosynthesis in the plant Arabidopsis thaliana. AB - Among the amino acids produced by plants cysteine plays a special role as a mediator between assimilatory sulfate reduction and provision of reduced sulfur for cell metabolism. Part of this characteristic feature is the presence of cysteine synthesis in plastids, mitochondria and cytosol. Plants are the major source of reduced sulfur for human and animal nutrition. Cysteine biosynthesis deserves special attention, since reduced sulfur is channelled from cysteine into many sulfur-containing compounds in food and feed. Recent investigations are reviewed that focus on structure and regulation of cysteine synthesis in the model plant Arabidopsis thaliana. These data indicate that cysteine synthesis is not just an intermediate reaction step but that it is part of a regulatory network that mediates between inorganic sulfur supply and the demand for reduced sulfur during plant growth and in response to environmental changes. PMID- 12083069 TI - Engineering of cysteine and methionine biosynthesis in potato. AB - Methionine and cysteine, two amino acids containing reduced sulfur, are not only an important substrate of protein biosynthesis but are also precursors of various other metabolites such as glutathione, phytochelatines, S-adenosylmethionine, ethylene, polyamines, biotin, and are involved as methyl group donor in numerous cellular processes. While methionine is an essential amino acid due to an inability of monogastric animals and human beings to synthesise this metabolite, animals are still able to convert methionine consumed with their diet into cysteine. Thus, a balanced diet containing both amino acids is necessary to provide a nutritionally favourable food or feed source. Because the concentrations of methionine and cysteine are often low in edible plant sources, e.g. potato, considerable efforts in plant breeding and research have been and are still performed to understand the physiological, biochemical, and molecular mechanisms that contribute to their synthesis, transport, and accumulation in plants. During the last decade molecular tools have enabled the isolation of most of the genes involved in cysteine and methionine biosynthesis, and the efficient plant transformation technology has allowed the creation of transgenic plants that are altered in the activity of individual genes. The physiological analysis of these transgenic plants has contributed considerably to our current understanding of how amino acids are synthesised. We focused our analysis on potato (Solanum tuberosum cv. Desiree) as this plant provides a clear separation of source and sink tissues and, for applied purposes, already constitutes a crop plant. From the data presented here and in previous work we conclude that threonine synthase and not cystathionine gamma-synthase as expected from studies of Arabidopsis constitutes the main regulatory control point of methionine synthesis in potato. This article aims to cover the current knowledge in the area of molecular genetics of sulfur-containing amino acid biosynthesis and will provide new data for methionine biosynthesis in solanaceous plants such as potato. PMID- 12083070 TI - Biosynthesis and metabolic engineering of glucosinolates. AB - Glucosinolates are amino acid-derived natural plant products found throughout the Capparales order. Glucosinolates and their degradation products have a wide range of biological activities, e.g. in plant defense as deterrents against insect and fungi. The conversion of amino acids to aldoximes is a key step in glucosinolate biosynthesis. This step is catalyzed by cytochromes P450 from the CYP79 family. The post-aldoxime enzymes in the glucosinolate pathway have high substrate specificity for the functional group and low substrate-specificity for the side chain. Therefore, we have been able to metabolically engineer new glucosinolate profiles into Arabidopsis by altering the levels of endogenous CYP79s and by introducing exogenous CYP79s. The approach has great potential for design of metabolically engineered plants with improved pest resistance and increased nutritional value. PMID- 12083071 TI - Research authority asked for HEW. PMID- 12083072 TI - A medical ethics issues survey of residents in five pediatric training programs. AB - Few pediatric training programs offer formal instruction or have ethics consultants to assist residents with ethical dilemmas. Moreover, even if such assistance were available, it is not clear that educators have an adequate understanding of (1) the range and frequency of possible ethical dilemmas residents may encounter, (2) residents' most troublesome concerns, (3) their use of resources to resolve issues, and (4) their own ethics belief systems. A sample of convenience (51 residents) at five midsouth residency programs was queried during 25-minute open-ended interviews to answer ethical questions; there were no refusals to participate. The "most troublesome" cases cited by the residents were related to life-and-death issues (withholding and withdrawing life support), child abuse and neglect cases, and disputes regarding patient care that arise between services. Two thirds of those surveyed indicated that they are still somewhat troubled by these difficult ethical problems. Surprisingly, residents stated that they relied on their peers more often than their attending physicians for effective assistance in resolving their most troublesome dilemmas. We support continuing education and research efforts to help residents and educators feel "more comfortable" in resolving ethical dilemmas. PMID- 12083073 TI - Physicians, patients, and decisions. PMID- 12083074 TI - Of morals and mercy. PMID- 12083075 TI - Code of professional deontology for medical and paramedical personnel at Catholic healthcare institutions. PMID- 12083077 TI - The Infant Life (Preservation) Act after fifty years. PMID- 12083076 TI - Abortion after ten years. PMID- 12083078 TI - Implications of the Plant Patent Act for the patentability of microorganisms. PMID- 12083079 TI - Laetrile: may the state intervene on behalf of a minor? PMID- 12083080 TI - Constitutional law--the rights of involuntarily committed mentally retarded persons under the Fourteenth Amendment: Youngberg v. Romeo. PMID- 12083081 TI - The recombinant DNA debate. [(Title provided[. PMID- 12083082 TI - The "right" to an abortion, the scope of Fourteenth Amendment "personhood," and the Supreme Court's birth requirement. PMID- 12083083 TI - Legal problems of surrogate motherhood. PMID- 12083084 TI - Due process for minors "voluntarily" committed to mental institutions: does father know best? Recommendations for Illinois. PMID- 12083085 TI - The genetic adventure. PMID- 12083086 TI - Eight worries about patenting animals. PMID- 12083087 TI - The born alive doctrine: a legal anachronism. PMID- 12083088 TI - Behavior modification: legal and ethical implications for the incarcerated. PMID- 12083089 TI - Constitutionally guaranteed rights of the mentally ill. PMID- 12083090 TI - Medical experiments and the new law: in vitro fertilisation procedures, embryo experimentation and the law. PMID- 12083091 TI - The abortion dilemma: twelve reports on a growing worldwide controversy. PMID- 12083092 TI - Souls on ice. PMID- 12083093 TI - Prenatal drug exposure: the constitutional implications of three governmental approaches. PMID- 12083094 TI - "In God we trust": when parents refuse medical treatment for their children based upon their sincere religious beliefs. PMID- 12083095 TI - Planned Parenthood v. Casey: the death of repose in reproductive decisionmaking. PMID- 12083096 TI - Patents on microorganisms. PMID- 12083097 TI - Common sense and the uncommon bacterium...is "life" patentable? PMID- 12083098 TI - A probabilistic justification for abortion. PMID- 12083099 TI - A study of the moral methodologies of intensive care units. PMID- 12083101 TI - Worthless life. PMID- 12083100 TI - Health care rationing and professional integrity: not necessarily incompatible. PMID- 12083102 TI - The health care proxy: ethical and policy choices in New York State. PMID- 12083103 TI - Ethics and AIDS. PMID- 12083104 TI - Medical ethics, medical economics and medical savings accounts. PMID- 12083105 TI - Legalistic and contextual approaches to living wills. PMID- 12083106 TI - What's distinctive about feminist bioethics? PMID- 12083107 TI - Are patients in a persistent vegetative state alive or dead? PMID- 12083108 TI - Proportional coinsurance: a market-based, ethically sound alternative to medical saving accounts. PMID- 12083110 TI - Historical relativism in bioethics: can we judge the standards and conduct of those who preceded us? PMID- 12083109 TI - Ethical issues in family medicine. PMID- 12083111 TI - Organizational ethics and health care providers. PMID- 12083112 TI - Seven (more) caveats concerning the discussion of euthanasia in the Netherlands. PMID- 12083113 TI - Arresting the white death: preventive detention, confinement for treatment, and medical ethics. PMID- 12083114 TI - Does a person have a right to die? PMID- 12083115 TI - Judaism and modern medical problems. PMID- 12083117 TI - Privacy and AIDS. PMID- 12083116 TI - Abortion/euthanasia. PMID- 12083118 TI - Laws and policies affecting fertility: a decade of change. PMID- 12083119 TI - Paradox: confidentiality/member of the team. PMID- 12083120 TI - The pregnant patient's bill of rights. PMID- 12083121 TI - Ethics and morals in nursing. PMID- 12083122 TI - Self-discipline or self-deception? PMID- 12083123 TI - The regulation of genetic engineering. PMID- 12083124 TI - Commentary: torture and the ethics of medicine. PMID- 12083125 TI - The law, the psychiatrist, and the patient. PMID- 12083126 TI - Distributing the burden fairly: ethics and national health policy. PMID- 12083127 TI - Human-semen banking: social and public policy issues. PMID- 12083128 TI - On abandoning life support: an alternative proposal. PMID- 12083129 TI - Williams Obstetrics on abortion. PMID- 12083130 TI - Ethical issues in behavior control: a preliminary examination. PMID- 12083131 TI - Recombinant DNA: triumph or Trojan horse? PMID- 12083132 TI - Regulating recombinant DNA research: pulling back from the apocalypse. PMID- 12083133 TI - Folio for folly: N.I.H. Guidelines for recombinant DNA research. PMID- 12083134 TI - A few remarks regarding research on recombinant DNA. PMID- 12083135 TI - Misuse of randomized clinical trials. PMID- 12083136 TI - The compliant client. PMID- 12083137 TI - Value conflicts and the uses of research: the example of abortion. PMID- 12083138 TI - The right to live and the right to die: some considerations of law and society in America. PMID- 12083139 TI - Sin boldly: ethical issues of DRGs. AB - The current system of diagnostic related groups poses a series of ethical concerns for patients, practitioners, and hospital administrators. In addition to reducing access to health care by those groups most in need, the system results in diminished quality, health care rationing based on cost/benefit projections, and en eventual two-tiered method of health care delivery. Most important, the author suggests that a radical change is occurring in the image of health care from service/mission to production/business with concomitant attitudinal changes in staff and administration. Capitulation to the business ethic results in the devaluation of "patients as products." Guidelines are offered to continue prudent fiscal management with service-oriented delivery to recover the historical mission of health care provision in hospitals. PMID- 12083141 TI - Medical and prescription records--patient access and confidentiality. PMID- 12083140 TI - Closing unprofitable services: ethical issues and management responses. AB - Closing unprofitable services often requires as much analysis, public relations, marketing, and planning as any expansion. Further, issues about ethics, indigents, and the hospital mission force the consideration of values explicitly if a marketing fiasco is to be avoided. By integrating values analysis with more traditional management tasks, the challenges of service closure can be converted into opportunities to demonstrate how your institution has met or exceeded its ethical obligations. A case involving OB is developed to show how ethical and management issues blend into one another. Specific strategies for consensus building and marketing of the legitimacy of the hospital's position are given. Institutional ethics committees are one primary mechanism for developing a plan to benefit from unpleasant decisions. PMID- 12083142 TI - The pharmacist and the patients' rights movement. PMID- 12083143 TI - Confidentiality--the psychotherapist's nemesis. PMID- 12083144 TI - Laetrile: should the dying patient decide? PMID- 12083145 TI - Moral integrity for nurses. PMID- 12083146 TI - Age and assisted reproduction. PMID- 12083147 TI - What is Jewish medical ethics? Arba'a Turim of Jacob ben Asher on medical ethics. PMID- 12083148 TI - In-vitro fertilization, embryo transfer, and embryo freezing. PMID- 12083149 TI - Religious traditions and public policy. PMID- 12083150 TI - Medicine and Judaism: an overview. PMID- 12083151 TI - Some modern responsa on medico-moral problems. PMID- 12083153 TI - AZT price cut for Third World mothers-to-be. PMID- 12083154 TI - The concept of precedent autonomy. AB - Does respect for autonomy imply respect for precedent autonomy? The principle of respect for autonomy requires us to respect a competent patient's treatment preference, but not everyone agrees that it requires us to respect preferences formed earlier by a now-incapacitated patient, such as those expressed in an advance directive. The concept of precedent autonomy, which concerns just such preferences, is problematic because it is not clear that we can still attribute to a now-incapacitated patient a preference which that patient never disaffirmed but can no longer understand. If we cannot make that attribution, then perhaps we should not respect precedent autonomy--after all, how can you respect patient autonomy by giving patients what they no longer want, even if they never disaffirmed those wants? I argue that whether an earlier preference can still be attributed to a now-incapacitated patient depends on the reasons behind the preference, for a preference includes (and is not merely supported by) the reasons behind it. When the considerations that served as reasons no longer exist, neither does the preference which included those reasons. In particular, if the considerations that served as reasons for the patient exist only under conditions where the patient retains full mental capacity, then once that capacity is lost, so are those reasons and the preference based upon them. I use this analysis of precedent autonomy to ascertain the merits of various approaches to advance medical decisionmaking, including Nancy Rhoden's approach, approaches based on a Parfitian personal identity analysis, approaches based on soft paternalism, and approaches based on the stability and longevity of preferences. Despite the apparent absurdity of respecting patient autonomy by giving patients what they no longer prefer but have never disaffirmed, I conclude with some programmatic remarks on when and why respect for (precedent) autonomy nonetheless requires us to respect former preferences. PMID- 12083156 TI - The ethics of euthanasia: advocates' perspectives. AB - The Netherlands is currently the only country in the world in which euthanasia is legally permissible. More specifically, Dutch law (briefly explained) allows that a doctor terminates the life of a patient of hers on his voluntary, well considered and sustained request, if he is suffering unbearably and hopelessly. The aim of this paper is to reconstruct the Dutch debate on the moral permissibility of euthanasia so as to clarify and strengthen the various views that can be advanced in support of euthanasia. On the one view, The Pure Autonomy View (TPAV), the justification of euthanasia rests solely on the principle of respect for autonomy. That is, the reason for performing and permitting euthanasia is the patient's voluntary, well-considered and sustained, in one word: autonomous, request for euthanasia. On the alternative view, The Joint View (TJV), the principle of respect for autonomy and the principle of beneficence morally justify euthanasia together. That is, euthanasia is ethical if and partly because, since the patient is suffering unbearably and hopelessly, euthanasia is in his interest. According to this paper, there is no easy argument for one of these views rather than the other. Instead, as yet both TPAV and TJV seem inherently problematic. TPAV is unable to give a doctor a reason for performing euthanasia that appeals to her in her capacity as a doctor, such as relief of suffering. And TJV begs the question--for example, if a state were to legalize euthanasia on grounds of TJV, it would force the view upon its citizens that it may be in a person's interest to die. PMID- 12083157 TI - What can the social sciences contribute to the study of ethics? Theoretical, empirical and substantive considerations. AB - This article seeks to establish that the social sciences have an important contribution to make to the study of ethics. The discussion is framed around three questions: (i) what theoretical work can the social sciences contribute to the understanding of ethics? (ii) what empirical work can the social sciences contribute to the understanding of ethics? And (iii) how does this theoretical and empirical work combine, to enhance the understanding of ethics, as a field of analysis and debate, is socially constituted and situated? Through these questions the argument goes beyond the now commonly cited objection to the over simplistic division between normative and descriptive ethics (that assigns the social sciences the 'handmaiden' role of simply providing the 'facts'). In extending this argument, this article seeks to establish, more firmly and in more detail, that: (a) the social sciences have a longstanding theoretical interest analysing the role that a concern with ethics plays in explanations of social change, social organisation and social action; (b) the explanations that are based on the empirical investigations conducted by social scientists exemplify the interplay of epistemological and methodological analyses so that our understanding of particular substantive issues is extended beyond the conventional questions raised by ethicists, and (c) through this combination of theoretical and empirical work, social scientists go beyond the specific ethical questions of particular practices to enquire further into the social processes that lie behind the very designation of certain matters as being 'ethical issues.' PMID- 12083155 TI - Reconsidering cochlear implants: the lessons of Martha's Vineyard. AB - I distinguish and assess three separate arguments utilized by the opponents of cochlear implants: that treating deafness as a medical condition is inappropriate since it is not a disability; that so treating it sends a message to the Deaf that they are of lesser worth; and that the use of such implants would signal the end of Deaf culture. I give some qualified support to the first and second claim, but find that the principal weight of the argument must be borne by the third argument: that use of the cochlear implants is impermissible because Deaf culture is intrinsically valuable. I show that this claim is, in practice, incompatible with the claim that deafness is not a disability: that the significant disadvantages suffered by the hearing impaired can only be corrected by measures that would end Deaf culture. Since the potential recipients of cochlear implants are, in the main, the prelingually deaf children of hearing parents, the burden of banning the implants would be borne by people who are not members of Deaf culture, and who owe that culture nothing over and above what we all owe cultures in general. I conclude that we cannot ask the parents of these children to sacrifice the interests of their children for the sake of Deaf culture. PMID- 12083158 TI - The politics of ethical consensus finding. PMID- 12083159 TI - Average wholesale price for prescription drugs: is there a more appropriate pricing mechanism? AB - This paper defines the average wholesale price (AWP), which has become an important benchmark for prescription drug pricing and reimbursement.The paper briefly explains the AWP's various uses in the pricing of prescription drugs, highlights some of the problems that have emerged as a result of the way it is reported and used, and explores some of the possibilities for reform. The paper also contains a glossary of commonly used terms, as well as an appendix that lists the state Medicaid reimbursement formulas. PMID- 12083160 TI - Will the nation be ready for the next bioterrorism attack? Mending gaps in the public health infrastructure. AB - This paper provides an overview of critical weaknesses in existing public health preparedness capabilities and discusses current policy initiatives to address these shortcomings. It examines developmental needs related to communication and coordination, information systems, laboratories, the development and distribution of vaccines and other countermeasures, emergency medical preparedness and response, and the public health workforce. The paper summarizes the current status of federal and state plans to respond to these developmental needs and touches on the future challenges likely to emerge as these plans are implemented. PMID- 12083161 TI - Poison control centers save lives and money. PMID- 12083162 TI - Summary of notifiable diseases--United States, 2000. AB - The MMWR Summary of Notifiable Diseases, United States, 2000 contains, in tabular and graphical form, the official statistics for the reported occurrence of nationally notifiable diseases in the United States for 2000. These statistics are collected and compiled from reports to the National Notifiable Diseases Surveillance System (NNDSS), which is operated by CDC in collaboration with the Council of State and Territorial Epidemiologists (CSTE). PMID- 12083164 TI - Hazards associated with using farm tractors to move large bales. PMID- 12083163 TI - Control of silica exposure from hand tools in construction: grinding concrete. PMID- 12083165 TI - Grain silo cleanup operation leads to two occupational deaths. PMID- 12083166 TI - Ergonomics demonstration project: skilled nursing facility. PMID- 12083167 TI - Estimating and controlling workplace risk: an approach for occupational hygiene and safety professionals. AB - The protection of people and physical assets is the objective of health and safety professionals and is accomplished through the paradigm of anticipation, recognition, evaluation, and control of risks in the occupational environment. Risk assessment concepts are not only used by health and safety professionals, but also by business and financial planners. Since meeting health and safety objectives requires financial resources provided by business and governmental managers, the hypothesis addressed here is that health and safety risk decisions should be made with probabilistic processes used in financial decision-making and which are familiar and recognizable to business and government planners and managers. This article develops the processes and demonstrates the use of incident probabilities, historic outcome information, and incremental impact analysis to estimate risk of multiple alternatives in the chemical process industry. It also analyzes how the ethical aspects of decision-making can be addressed in formulating health and safety risk management plans. It is concluded that certain, easily understood, and applied probabilistic risk assessment methods used by business and government to assess financial and outcome risk have applicability to improving workplace health and safety in three ways: 1) by linking the business and health and safety risk assessment processes to securing resources, 2) by providing an additional set of tools for health and safety risk assessment, and 3) by requiring the risk assessor to consider multiple risk management alternatives. PMID- 12083168 TI - A simple method for tracer containment testing of hospital isolation rooms. AB - A simple method for tracer containment testing of hospital isolation rooms is presented. The method does not require any equipment setup in tested rooms and can be completed in approximately one-half hour per room. Tracer samples are taken at specified time intervals in the corridor outside of an isolation room and analyzed on a portable gas chromatograph system. Results are presented from tracer testing of two isolation rooms in two different hospitals. One isolation room had a significant negative pressure differential between room and corridor, and the other isolation room was not at negative pressure. A small quantity of sulfur hexafluoride gas was injected manually from a polyethylene syringe over a bed in an isolation room. Tracer concentrations were thereafter measured in the corridor adjacent to the room at 5-minute intervals for 20 minutes after the injection, yielding a quantitative measure of leakage of the tracer from the isolation room. Finally, measuring the tracer concentration in the isolation room 30 minutes after injection yielded an indication of how effectively the ventilation system removed a contaminant released at the position of the bed. The results show that an instantaneous release of a small quantity of tracer gas in an isolation room yields tracer concentrations in the corridor outside of the room that are within the analytical range of the measuring equipment both for a properly functioning isolation room and an improperly functioning isolation room, and thus that the method is well-suited for studying containment in hospital isolation rooms. Possible practical applications of the method are discussed. PMID- 12083169 TI - Diisocyanate emission from a paint product: a preliminary analysis. AB - Exposure of workers to diisocyanates in the polyurethane foam manufacturing industry is well documented. However, very little quantitative data have been published on exposure to diisocyanates from the use of paints and coatings. The purpose of this study was to evaluate emission of 2,4-toluene diisocyanate, 2,6 toluene diisocyanate (2,6-TDI), and isophorone diisocyanate from a commercially available two-stage concrete coating and sealant. A laboratory model of an outdoor deck coating process was developed and diisocyanate concentrations determined by derivatization with 1-(2-methoxyphenol)-piperazine and subsequent high performance liquid chromatographic analysis with UV detection. The detection limit for 2,4-toluene diisocyanate and 2,6-toluene diisocyanate urea derivatives was 0.6 microg TDI/gm wet product, and 0.54 microg IPDI/gm wet product for the isophorone diisocyanate urea derivative. No 2,4-toluene diisocyanate or isophorone diisocyanate was detected in the mixed product. A maximum mean 2,6-TDI emission rate of 0.32 microg of 2,6-TDI/gram of wet product applied/hour was observed for the 1-hour sampling time, 0.38 microg of 2,6-TDI/gram of wet product applied/hour was observed for the 5-hour sampling time, and 0.02 micrpg of 2,6 TDI/gram of wet product applied/hour was observed for the 15-hour sampling time. The decrease in rate of 2,6-TDI emission over the 15-hour period indicates that emission of 2,6-TDI is virtually complete after 5 hours. These emission rates should allow industrial hygienists to calculate exposures to isocyanates emitted from at least one curing sealant. PMID- 12083170 TI - A spectrophotometric method for the determination of organic soluble matter in bitumen fumes. AB - A UV spectrophotometric procedure was validated for the determination of organic soluble matter in bitumen fumes collected by filtration technique. Ultrasonic extraction was carried out with toluene, an efficient extraction solvent for polycyclic aromatic hydrocarbons, followed by UV absorbance measurements at 320 nm. A calibration curve is plotted from the same set of samples determined by classical weighing method. Further determinations can also be made using the slope factor of the calibration curve. The procedure presents obvious simplicity and rapidity advantages and is less prone to losses than the measurements of weight. Inter-method comparisons of samples collected from experimental laboratory-generated penetration bitumen fumes commonly used in road paving showed that the three available procedures-weighing, infrared, and UV--described for the determination of organic soluble matter yield equivalent results. PMID- 12083171 TI - Occupational carbon monoxide violations in the State of Washington, 1994-1999. AB - Occupational exposure to carbon monoxide continues to cause a number of injuries and deaths. This study reviewed the State of Washington OSHA inspection records for occupational safety or health violations related to carbon monoxide for the time period 1994-1999 to assess the agency's efforts and further identify and characterize causative factors. Inspection data were also compared with carbon monoxide claims data from a companion study to determine if the agency was visiting the most at risk work operations. Inspections were identified by searching computerized violation texts for "carbon monoxide" or "CO." The study found 142 inspections with one or more carbon monoxide violations. Inspections were spread over 84 different 4-digit Standard Industrial Classification codes. Most inspections were initiated as a result of a complaint or other informant. Inspections were predominantly in construction and manufacturing, whereas carbon monoxide claims were mores evenly distributed between the major industries. Inspections also may have failed to find violations for some types of equipment responsible for carbon monoxide claims. Forklifts were the source of carbon monoxide most often associated with a violation, followed by compressors for respirators, auto/truck/bus, and temporary heating devices. Inspections in response to poisonings found common factors associated with lack of recognition and failure to use or maintain equipment and ventilation. Some work sites with one or more poisonings were not being inspected. Only 10 of the 51 incidents with industrial insurance claim reports of carboxyhemoglobin at or above 20 percent were inspected. Further, it was found more preventive efforts should be targeted at cold storage operations and certain warehouse and construction activities. It is proposed that more specific standards, both consensus and regulatory, would provide additional risk reduction. Reliance upon safe work practices as a primary method of control in the use of fuel-powered equipment in cold storage or other enclosed and unventilated environments needs to be prohibited. The study further demonstrates how inspection and industrial insurance records can assist with preventive efforts and better focus an agency's efforts. PMID- 12083173 TI - Emotional intelligence: a core competency for health care administrators. AB - The contemporary concept of emotional intelligence (EI) as a critical set of management skills is traced through time to its current application for health care administration. EI is defined as proficiencies in intrapersonal and interpersonal skills in the areas of self-awareness, self-regulation, self motivation, social awareness, and social skills. The contributions of EI to effective management are supported by empirical research in the field. The importance of developing these skills in health care organizations is further clarified with examples familiar to health care administrators. Training suggestions and assessment resources are provided. PMID- 12083172 TI - Thermal loading as a causal factor in exceeding the 0.1 PPM laboratory fume hood control level. AB - Tracer gas testing per ANSI/ASHRAE 110-1995 Method of Testing Performance of Laboratory Fume Hoods was used to investigate the role of thermal loading in exceeding laboratory fume hood control levels. Three types of typical laboratory burners (blast, Meeker, and economy) were used to provide a thermal challenge. Heat outputs of between 0 and 61,610 Btu/hr were based on fuel heat capacity (for liquid propane gas) and fuel gas flow rates. Breathing zone concentrations were measured with a MIRAN 1B2 infrared gas analyzer. Also, for each test, the difference between the room and duct temperatures (delta temperature) was measured. Results indicated a linear relationship between heat loads and tracer gas breathing zone concentrations for both Btu/hr and delta temperature. Control levels of 0.1 ppm were exceeded at less than 12,000 Btu/hr. Also, control levels were exceeded at a lower heat load when the tracer gas generation rate was increased. These results indicate that thermal loads in laboratory fume hoods increase the risk of exceeding laboratory fume hood control levels. Some compensatory measures relative to hood configuration and flow rates are recommended for laboratory operations involving heat sources. PMID- 12083174 TI - Communication: bridging the racial and ethnic divide in health care management. AB - The emerging body of literature on racial and ethnic diversity in health services management reveals four pervasive and formidable challenges for health care industry leadership. These are: under-representation of people of color in health services management, persistent gaps in both compensation and satisfaction between managers of color and white managers, and racial and ethnic disparities in medical care process and outcomes. Focus groups of health care managers were assembled to explore the intersection of race, ethnicity, and health care management. Results of the focus group discussions support the need for meaningful dialogue that aims to identify common ground and shared purpose so that appropriate actioncan be taken to bridge the racial and ethnic divide. PMID- 12083175 TI - How physical therapists perceive physicians: a stakeholder analysis. AB - Physicians and physical therapy service units are facing a different health care climate than 5 to 10 years ago. Managed care, changes in Medicare reimbursement, and other financing issues are causing turbulence for the industry. This article reports on more than 1,300 physical therapy service unit managers of hospitals, long-term care facilities, outpatient clinics, and home health agencies who determined the most important stakeholderoftheir physical therapy service unit and the perceived degree of power the various stakeholders had over the physical therapy service area. This study indicates that historical roles and alliances in health care are undergoing major alterations. PMID- 12083176 TI - The impact of union elections on human resources management practices in hospitals. AB - The purpose of this article is to explore top management's perceptions of how various human resources management (HRM) practices changed in hospitals (n = 101) after union elections. Significant increases in many HRM practices that are believed to lead to competitive advantage through human resources were reported in firms in which unions lost elections but not in firms where unions were certified. PMID- 12083177 TI - Understanding the members of your health care "gang:" seekers and cruisers. AB - Managers are challenged by diverse characteristics of persons belonging to certain groups. This paper outlines characteristics of "cruisers" and "seekers" and managerial strategies for responding to these characteristics for leadership effectiveness. PMID- 12083178 TI - Sexual harassment in the health care industry: a follow-up inquiry. AB - A nationwide survey of hospital human resources managers reveals that allegations of sexual harassment in hospitals are increasing. Reported statistics for a 4.5 year period show that nurses continue to bring the largest number of charges. Most are "hostile environment" allegations, and most formal charges are levied against coworkers. When compared to data gathered in an earlier survey, these statistics show an alarming trend. A significant increase in allegations occurred in 1999 and 2000, corresponding to two US Supreme Court rulings that clarify an employer's responsibility to eliminate this form of sexual discrimination from the workplace. PMID- 12083179 TI - Case management model or case manager type? That is the question. AB - Case management models have been categorized many different ways, but classification into two generic models is proposed here: an interrogative model that relies on intense oversight with expected cost reduction and a patient advocacy model that relies on a brokering arrangement for services in the best interest of the patient. A randomized trial in an elderly, functionally impaired population resulted in the implementation of a patient advocacy model and suggests that this model results in increased service use and costs, but that increased survival rates may justify the additional cost. PMID- 12083181 TI - A manager asks: Dealing with the troublesome employee. PMID- 12083180 TI - Making change work for you--or at least not against you. AB - Resistance is an automatic response by most people--managers as well as nonmanagerial employees--to change that is thrust on them. Resistance arises in large part from a feeling of helplessness in the presence of that which the individual cannot control. Change is resisted by most people because of centuries of conditioning that taught them to seek security in constancy. Today's security, however, more likely lies in flexibility and adaptability. The department manager has the difficult task of maintaining a positive attitude in the face of change, accepting that which cannot be controlled and providing a positive role model for employees. Although one may feel helpless in the presence of change, there is always part of the circumstances one can influence or control. People make change work for themselves by accepting what they cannot control and taking ownership of what they can control. PMID- 12083183 TI - Articulation index predictions for hearing-impaired listeners with and without cochlear dead regions. PMID- 12083185 TI - Analysis of one-dimensional wave scattering by the random walk method. AB - The random walk method is applied to a one-dimensional Helmholtz equation with a variable wave number. The solution is represented as a mathematical expectation of a specified functional on paths in a complex space. This solution degenerates to the ray-method approximation in domains where the latter method may be used, but the probabilistic formulas presented describe also backscattered waves whose existence is not explained by the standard asymptotic techniques. The numerical results confirm the efficiency of the random walk approach to the analysis of wave propagation. PMID- 12083184 TI - Effect of duration on amplitude-modulation masking. PMID- 12083186 TI - Reflection of drill-string extensional waves at the bit-rock contact. AB - Downward propagating extensional waves are partially reflected at the bit-rock contact. The evaluation of the reflection coefficient is important to obtain while drilling information about the acoustic properties of the formations. The scope of this work is to estimate the bit-rock reflection coefficient, assuming a flat drill bit in perfect contact with the formation. Using the low-frequency approximation, which holds when the wavelength is much larger than the lateral dimensions of the borehole, the drill-string is assumed to be a laterally free rod, and the formation an homogeneous and isotropic medium. This work shows that the reflection coefficient of the extensional waves depends, along with the elastic properties of the formation, on the ratio of the cross sections of the drill-string and borehole. The impedance of the drilled rock can be calculated from the measured reflection coefficient, which is related to the amplitude of waves produced in the string and in the formation by a working drill-bit. PMID- 12083187 TI - A direct mixed-body boundary element method for packed silencers. AB - Bulk-reacting sound absorbing materials are often used in packed silencers to reduce broadband noise. A bulk-reacting material is characterized by a complex mean density and a complex speed of sound. These two material properties can be measured by the two-cavity method or calculated by empirical formulas. Modeling the entire silencer domain with a bulk-reacting lining will involve two different acoustic media, air and the bulk-reacting material. Traditionally, the interior silencer domain is divided into different zones and a multi-domain boundary element method (BEM) may be applied to solve the problem. However, defining different zones and matching the elements along each interface is tedious, especially when the zones are intricately connected. In this paper, a direct mixed-body boundary element method is used to model a packed silencer without subdividing it into different zones. This is achieved by summing up all the integral equations in different zones and then adding the hypersingular integral equations at interfaces. Several test cases, including a packed expansion chamber with and without an absorbing center bullet, and a parallel baffle silencer, are studied. Numerical results for the prediction of transmission loss (TL) are compared to experimental data. PMID- 12083188 TI - Scattering by a fluid cylinder in a porous medium: application to trabecular bone. AB - In a trabecular bone, considered as a nondissipative porous medium, the scattering of an incident wave by cylindrical pores larger than the wavelength is studied. The goal is to know if scattering alone may cause such a high attenuation as that observed in calcaneus. The porous medium is modelized via Biot's theory and the scattering by a single pore is characterized from the definition of a scattering matrix. An approximation of weakly disordered medium is then discussed to estimate the effective attenuation and dispersion as a function of frequency. These effective properties are shown to be different of those measured on calcaneus, due to the neglect of wave conversions during the scattering process. PMID- 12083189 TI - Characterizing holes in duct walls using resonance frequencies. AB - The current paper demonstrates that the technique recently used by de Salis and Oldham for the sizing and location of blockages in ducts using resonance and antiresonance value shifts [J. Sound Vib. 221(1), 180-186 (1999)] may be successfully applied to the detection, location, and sizing of small holes in duct walls. It is shown that when the blockage area function reconstruction process is applied using resonance and antiresonance frequencies determined from a single measurement of sound pressure within a duct, the position of a hole in the duct wall is revealed as the beginning of an apparent gradual expansion. This expansion is termed the effective expansion function and emanates from an initial positive dc shift. Once the hole has been located, a simple impedance model of the duct incorporating the hole location and the measured wave number at the first-order duct resonance may be derived which allows the determination of the radius of the side hole from a simple quadratic equation. This hole-sizing technique is also successfully applied to thin slits in duct walls using an equivalent radius approach, and is shown to be highly accurate for slit-shaped holes with cross-sectional length to width aspect ratios of up to 28:1. PMID- 12083190 TI - Implosion of an underwater spark-generated bubble and acoustic energy evaluation using the Rayleigh model. AB - The growth, collapse, and rebound of a vapor bubble generated by an underwater spark is studied by means of high-speed cinematography, simultaneously acquiring the emitted acoustic signature. Video recordings show that the growth and collapse phases are nearly symmetrical during the first two or three cycles, the bubble shape being approximately spherical. After 2-3 cycles the bubble behavior changes from a collapsing/rebounding regime with sound-emitting implosions to a pulsating regime with no implosions. The motion of the bubble wall during the first collapses was found to be consistent with the Rayleigh model of a cavity in an incompressible liquid, with the inclusion of a vapor pressure term at constant temperature within each bubble cycle. An estimate of the pressure inside the bubble is obtained measuring the collapse time and maximum radius, and the amount of energy converted into acoustical energy upon each implosion is deduced. The resulting value of acoustic efficiency was found to be in agreement with measurements based on the emitted acoustic pulse. PMID- 12083191 TI - Numerical analysis of dipole sound source around high speed trains. AB - As the maximum speed of high speed trains increases, the effect of aeroacoustic noise on the sound level on the ground becomes increasingly important. In this paper, the distribution of dipole sound sources at the bogie section of high speed trains is predicted numerically. The three-dimensional unsteady flow around a train is solved by the large eddy simulation technique. The time history of vortices shows that unstable shear layer separation at the leading edge of the bogie section sheds vortices periodically. These vortices travel downstream while growing to finally impinge upon the trailing edge of the section. The wavelength of sound produced by these vortices is large compared to the representative length of the bogie section, so that the source region can be regarded as acoustically compact. Thus a compact Green's function adapted to the shape can be used to determine the sound. By coupling the instantaneous flow properties with the compact Green's function, the distribution of dipole sources is obtained. The results reveal a strong dipole source at the trailing edge of the bogie section where the shape changes greatly and the variation of flow with time is also great. On the other hand, the bottom of the bogie section where the shape does not change, or the leading edge and boundary layer where the variation of flow with time is small, cannot generate a strong dipole source. PMID- 12083192 TI - Time reversed reverberation focusing in a waveguide. AB - Time reversal mirrors have been applied to focus energy at probe source locations and point scatterers in inhomogeneous media. In this paper, we investigate the application of a time reversal mirror to rough interface reverberation processing in a waveguide. The method is based on the decomposition of the time reversal operator which is computed from the transfer matrix measured on a source-receiver array [Prada et al., J. Acoust. Soc. Am. 99, 2067-2076 (1996)]. In a similar manner, reverberation data collected on a source-receiver array can be filtered through an appropriate temporal window to form a time reversal operator. The most energetic eigenvector of the time reversal operator focuses along the interface at the range corresponding to the filter delay. It is also shown that improved signal-to-noise ratio measurement of the time reversal operator can be obtained by ensonifying the water column with a set of orthogonal array beams. Since these methods do not depend upon a priori environmental information, they are applicable to complex shallow water environments. Numerical simulations with a Pekeris waveguide demonstrate this method. PMID- 12083193 TI - Matched-field processing gain degradation caused by tidal flow over continental shelf bathymetry. AB - Temporally variable, range dependent sound-speed profiles measured during ebb flow and estimated for slack flow are used to quantify the variability of matched field signal-processing gain degradation in shallow water propagation channels controlled by tidally driven stratified flow over variable bathymetry. Calculations along a 9.3 km range establish phase changes in the acoustic signal as the primary cause of a 3-9 dB degradation in the coherent matched-field processing output of a full water column vertical array. The work indicates that over a tidal cycle acoustic signal properties and matched-field processing gain can be expected to change continuously in a shallow water stratified channel that has bathymetry variability. Acoustic signals propagating in such tidal flow controlled environments may be expected to display repeatable (over successive tidal cycles) and predictable changes in their phase coherent properties. These results suggest that matched-field processor replica fields used in the shelf/slope propagation environment will have to be updated regularly during a tidal cycle to maintain maximum processor gain. PMID- 12083194 TI - The high-frequency backscattering angular response of gassy sediments: model/data comparison from the Eel River Margin, California. AB - A model for the high-frequency backscatter angular response of gassy sediments is proposed. For the interface backscatter contribution we adopted the model developed by Jackson et al. [J. Acoust. Soc. Am. 79, 1410-1422 (1986)], but added modifications to accommodate gas bubbles. The model parameters that are affected by gas content are the density ratio, the sound speed ratio, and the loss parameter. For the volume backscatter contribution we developed a model based on the presence and distribution of gas in the sediment. We treat the bubbles as individual discrete scatterers that sum to the total bubble contribution. This total bubble contribution is then added to the volume contribution of other scatters. The presence of gas affects both the interface and the volume contribution of the backscatter angular response in a complex way that is dependent on both grain size and water depth. The backscatter response of fine grained gassy sediments is dominated by the volume contribution while that of coarser-grained gassy sediments is affected by both volume and interface contributions. In deep water the interface backscatter is only slightly affected by the presence of gas while the volume scattering is strongly affected. In shallow water the interface backscatter is severely reduced in the presence of gas while the volume backscatter is only slightly increased. Multibeam data acquired offshore northern California at 95 kHz provides raw measurements for the backscatter as a function of grazing angle. These raw backscatter measurements are then reduced to scattering strength for comparison with the results of the proposed model. The analysis of core samples at various locations provides local measurements of physical properties and gas content in the sediments that, when compared to the model, show general agreement. PMID- 12083195 TI - Single bubble sonoluminescence driven by non-simple-harmonic ultrasounds. AB - The dependence of the single bubble sonoluminescence (SBSL) on the waveforms of the driving ultrasound has been investigated by both experiment and numerical calculation. Three types of non-simple-harmonic waves, the rectangular, triangular and as well as the sinusoidal wave with a pulse, are used to drive the SBSL in our research. The triangular wave is the most effective, while the rectangular wave is the worst and the sinusoidal wave in the middle. However, the rectangular wave drives the brightest SBSL among those waves if the sound pressure amplitude keeps constant. When we use a simple-harmonic wave with a pulse as the driving sound, stable and periodic SBSL flashes have been observed. An increase in the flash intensity can be observed as the pulse is put at a suitable phase related to the sinusoidal wave. All of the observations are investigated numerically. Well qualitative agreements between the numerical simulations and the experimental measurements have been achieved. PMID- 12083196 TI - Application of narrow band laser ultrasonics to the nondestructive evaluation of thin bonding layers. AB - In this paper, a modified laser induced grating technique (LIG) has been utilized to generate narrow band surface waves in an epoxy-bonded copper-aluminum layered structure. A high performance optical interferometer system was utilized to detect the laser-generated surface waves. The dispersion of surface wave in an epoxy-bonded copper-aluminum specimen was measured and compared with the theoretical solution. An inverse algorithm based on the simplex method was then introduced to determine the bonding thickness as well as the elastic properties of the bonding layer. The inversion results demonstrated that the thickness in the microm range or the elastic properties of the bonding layer could be successfully determined. PMID- 12083197 TI - Determination of embedded layer properties using adaptive time-frequency domain analysis. AB - A general model for determination of the complete set of acoustical and geometrical properties of an isotropic layer embedded between isotropic or anisotropic multilayered solids is developed. These properties include density, longitudinal and shear elastic moduli, layer thickness, and loss factors, simultaneously determined from two measurements, one at normal and one at oblique incidence. The inversion model is an extension of the method proposed by Lavrentyev and Rokhlin [J. Acoust. Soc. Am. 102, 3467 (1997)] which is applicable to thick substrates. In this new method, the inversion model mimics an experiment by using the same time-domain gating of the signal reflected from the embedded layer. This allows application of this method to layered solids when reflections from different layers overlap in the time domain. The sensitivity of the method, its stability against data scatter, and the effect of the oblique incident angle are evaluated. The effect of plane-wave approximation versus beam approximation in the inverse algorithm is discussed. Experimental results are given to demonstrate examples of adhesive layer property reconstruction. PMID- 12083198 TI - Thermoacoustics with idealized heat exchangers and no stack. AB - A model is developed for thermoacoustic devices that have neither stack nor regenerator. These "no-stack" devices have heat exchangers placed close together in an acoustic standing wave of sufficient amplitude to allow individual parcels of gas to enter both exchangers. The assumption of perfect heat transfer in the exchangers facilitates the construction of a simple model similar to the "moving parcel picture" that is used as a first approach to stack-based engines and refrigerators. The model no-stack cycle is shown to have potentially greater inviscid efficiency than a comparable stack model. However, losses from flow through the heat exchangers and on the walls of the enclosure are greater than those in a stack-based device due to the increased acoustic pressure amplitude. Estimates of these losses in refrigerators are used to compare the possible efficiencies of real refrigerators made with or without a stack. The model predicts that no-stack refrigerators can exceed stack-based refrigerators in efficiency, but only for particular enclosure geometries. PMID- 12083199 TI - The acoustic emission of a distributed mode loudspeaker near a porous layer. AB - Experimental and theoretical modeling of the vibro-acoustic performance of a distributed mode loudspeaker (DML) suggest that their acoustic emission can be significantly affected by the presence of a porous layer. The amplitude of the surface velocity of the panel and the acoustic pressure on the porous surface are reduced largely in the vicinity of structural resonances due to the additional radiation damping and visco-thermal absorption phenomenon in the porous layer. The experimental results suggest that a porous layer between a rigid base and a DML panel can considerably alter its acoustic emission in the near field and in the far field. This is illustrated by a reduction in the level of fluctuations in the emitted acoustic pressure spectra. These fluctuations are normally associated with the interference between the sound emitted by the front surface of the speaker and that emitted from the back. Another contribution comes from the pronounced structural resonances in the surface velocity spectrum. The results of this work suggest that the acoustic boundary conditions near a DML can be modified by the porous layer so that a desired acoustic output can be attained. PMID- 12083200 TI - Generalization of a model of hysteresis for dynamical systems. AB - A previously described model of hysteresis [J. C. Piquette and S. E. Forsythe, J. Acoust. Soc. Am. 106, 3317-3327 (1999); 106, 3328-3334 (1999)] is generalized to apply to a dynamical system. The original model produces theoretical hysteresis loops that agree well with laboratory measurements acquired under quasi-static conditions. The loops are produced using three-dimensional rotation matrices. An iterative procedure, which allows the model to be applied to a dynamical system, is introduced here. It is shown that, unlike the quasi-static case, self-crossing of the loops is a realistic possibility when inertia and viscous friction are taken into account. PMID- 12083201 TI - An approximated 3-d model of the Langevin transducer and its experimental validation. AB - In this work, an approximated 3-D analytical model of the Langevin transducer is proposed. The model, improving the classical 1-D approach describing the thickness extensional mode, allows us to predict also the radial modes of both the piezoelectric ceramic disk and the loading masses; furthermore, it is able to describe the coupling between radial and thickness extensional modes. In order to validate the model, the computed frequency spectrum is compared with that obtained by measurements carried out on 13 manufactured samples of different thicknesses to diameter ratios. The comparison shows that the model predicts with quite good accuracy the resonance frequencies of the two lowest frequency modes, i.e., those of practical interest, all over the explored range. Finally, the coupling effect between thickness and radial modes on the frontal displacement is measured and discussed. PMID- 12083202 TI - Electrical characterization of plate piezoelectric transducers bonded to a finite substrate. AB - This article presents a new technique for characterizing piezoelectric transducers attached to a finite substrate. It consists of determining the impedance of the transducer cleared of the effects caused by finite dimensions of the substrate. This technique is validated by comparison with measurements on a transducer mounted on an effectively half-infinite substrate. It is applied for the electrical matching of a lithium niobate plate transducer bonded to a fused quartz rod. PMID- 12083203 TI - The analysis and interpretation of some special properties of higher order symmetric lamb waves: the case for plates. AB - Results derived from exact linear homogeneous elastodynamic theory are used for two-dimensional unloaded plates in order to understand certain features generated by proper symmetric Lamb modes. It is shown that S1 modes for all elastic materials have a phase velocity defined below the usual critical frequency and which initially exhibits anomalous dispersion (has a negative slope with respect to frequency). Over a certain range, it has a phase velocity that is double valued. In addition, there are an infinite number of proper symmetric Lamb modes that have this characteristic for materials with a Poisson ratio equal to 1/3. It also appears that all A3n modes are anomalous when V(L) < or = 2 V(T). The cause and implication of these effects are examined, including an associated negative group velocity over a small frequency zone for these modes. Further, it is noted that all proper symmetric Lamb modes have a plateau region in phase velocity with respect to wave number. It is shown that this always occurs for a phase velocity corresponding to the longitudinal bulk velocity of the elastic material. These issues are examined along with how one may obtain material parameters and possibly plate thickness from their dispersion curves. PMID- 12083204 TI - Free interfacial vibrations in cylindrical shells. AB - The 2D equations in the Kirchhoff-Love theory are subjected to asymptotic analysis in the case of free interfacial vibrations of a longitudinally inhomogeneous infinite cylindrical shell. Three types of interfacial vibrations, associated with bending, super low-frequency semi-membrane, and extensional motions, are investigated. It is remarkable that for extensional modes natural frequencies have asymptotically small imaginary parts caused by a weak coupling with propagating bending waves. Bending and extensional vibrations correspond to Stonely-type plate waves, while semi-membrane ones are strongly dependent on shell curvature and do not allow flat plate interpretation. The paper represents generalization of the recent authors' publication [Kaplunov et al., J. Acoust. Soc. Am. 107, 1383-1393 (2000)] dealing with edge vibrations of a semi-infinite cylindrical shell. PMID- 12083205 TI - Nonlinear seismo-acoustic land mine detection and discrimination. AB - A novel technique for detection and discrimination of artificial objects, such as land mines, pipes, containers, etc., buried in the ground, has been developed and tested. The developed approach utilizes vibration (using seismic or airborne acoustic waves) of buried objects, remote measurements of soil surface vibration (using laser or microwave vibrometers), and processing of the measured vibration to extract mine's "vibration signatures." The technique does not depend upon the material from which the mine is fabricated whether it be metal, plastic, wood, or any other material. It depends upon the fact that a mine is a "container" whose purpose is to contain explosive materials and associated detonation apparatus. The mine container is in contact with the soil in which it is buried. The container is an acoustically compliant article, whose compliance is notably different from the compliance of the surrounding soil. Dynamic interaction of the compliant container and soil on top of it leads to specific linear and nonlinear effects used for mine detection and discrimination. The mass of the soil on top of a compliant container creates a classical mass-spring system with a well defined resonance response. Besides, the connection between mass (soil) and spring (mine) is not elastic (linear) but rather nonlinear, due to the separation of the soil/mine interface in the tensile phase of applied dynamic stress. These two effects, constituting the mine's vibration signature have been measured in numerous laboratory and field tests, which proved that the resonance and nonlinear responses of a mine/soil system can be used for detection and discrimination of buried mines. Thus, the fact that the mine is buried is turned into a detection advantage. Because the seismo-acoustic technique intrinsically detects buried containers, it can discriminate mines from noncompliant false targets such as rocks, tree roots, chunks of metal, bricks, etc. This was also confirmed experimentally in laboratory and field tests. PMID- 12083206 TI - Coupling analysis of a matched piezoelectric sensor and actuator pair for vibration control of a smart beam. AB - This paper presents a theoretical and experimental study of the in-plane and out of-plane coupling of a matched piezoelectric sensor/actuator pair bonded on a beam. Both the sensor and actuator are triangularly shaped polyvinylidene fluoride (PVDF) transducers and are intended to provide a compact sensor/actuator system for beam vibration control. The measured sensor-actuator frequency response function has shown an unpredicted increase in magnitude with frequency, which was found, to be due to in-plane vibration coupling. An analytical model has been developed to decompose the sensor-actuator response function into an in plane contribution and an out-of-plane contribution. This in-plane coupling can limit the feedback control gains when a direct velocity feedback control is applied. A method called the j omega s compensation method is proposed to identify the effect of the in-plane vibration coupling at low frequencies. Even after this compensation, however, there was unexpected strong out-of-plane coupling at even modes, which may have been caused by a lack of accuracy in the shaping of the PVDF sensor and actuator. Numerical simulations have confirmed the sensitivity of the matched sensor/actuator pair with shaping errors. PMID- 12083207 TI - Robust active control of broadband noise in finite ducts. AB - This study focuses on robust active control of broadband noise in finite ducts. Our analytical and experimental studies suggest the existence of several technical flaws in the path models of conventional active noise control (ANC) systems. These are sensitivity of the path model with respect to boundary conditions, and nonminimum phase (NMP) secondary and reference paths. For finite ducts with small cross sections, the traveling wave model (TWM) may be adopted to find an effective solution to these problems and lead to a robust ANC system. Since many practical "noisy" ducts are finite with small cross sections, the proposed ANC has many practical applications. Its robustness and ability to suppress broadband noise will be explained theoretically and verified experimentally. PMID- 12083208 TI - Measured anisotropic air flow resistivity and sound attenuation of glass wool. AB - The air flow resistivity of glass wool has been measured in different directions. The glass wool was delivered from the manufacturer as slabs measuring 100x600x900 mm3, where the surface 600x900 mm2 was parallel with the conveyor belt used in the manufacturing. Directions in the glass wool are described by a coordinate system with the X axis perpendicular to the conveyor belt, the Z axis in the direction the conveyor belt moves, and the Y axis perpendicular to the two other axes. It was found that the resistivities in the Y and Z directions were equal in all cases. For density 14 kg/m3 the mean resistivity in the X direction was 5.88 kPa s m(-2) and in the Y direction 2.94 kPa s m(-2). For density 30 kg/m3 the mean resistivity in the X direction was 15.5 kPa s m(-2) and in the Y direction 7.75 kPa s m(-2). A formula for prediction of resistivity for other densities is given. By comparing measured values of sound attenuation with results calculated from resistivity data, it is demonstrated that the measured attenuation can be predicted in a simple manner. PMID- 12083210 TI - Interaction between adenosine triphosphate and mechanically induced modulation of electrically evoked otoacoustic emissions. AB - It was shown previously that electrically evoked otoacoustic emissions (EEOAEs) can be amplitude modulated by low-frequency bias tones and enhanced by application of adenosine triphosphate (ATP) to scala media. These effects were attributed, respectively, to the mechano-electrical transduction (MET) channels and ATP-gated ion channels on outer hair cell (OHC) stereocilia, two conductance pathways that appear to be functionally independent and additive in their effects on ionic current through the OHC. In the experiments described here, the separate influences of ATP and MET channel bias on EEOAEs did not combine linearly. Modulated EEOAEs increased in amplitude, but lost modulation at the phase and frequency of the bias tone (except at very high sound levels) after application of ATP to scala media, even though spectral components at the modulation sideband frequencies were still present. Some sidebands underwent phase shifts after ATP. In EEOAEs modulated by tones at lower sound levels, substitution of the original phase values restored modulation to the waveform, which then resembled a linear summation of the separate effects of ATP and low-frequency bias. While the physiological meaning of this procedure is not clear, the result raises the possibility that a secondary effect of ATP on one or more nonlinear stages in the transduction process, which may have caused the phase shifts, obscured linear summation at lower sound levels. In addition, "acoustic enhancement" of the EEOAE may have introduced nonlinear interaction at higher levels of the bias tones. PMID- 12083209 TI - One source for distortion product otoacoustic emissions generated by low- and high-level primaries. AB - Distortion product otoacoustic emissions (DPOAE) elicited by tones below 60-70 dB sound pressure level (SPL) are significantly more sensitive to cochlear insults. The vulnerable, low-level DPOAE have been associated with the postulated active cochlear process, whereas the relatively robust high-level DPOAE component has been attributed to the passive, nonlinear macromechanical properties of the cochlea. However, it is proposed that the differences in the vulnerability of DPOAEs to high and low SPLs is a natural consequence of the way the cochlea responds to high and low SPLs. An active process boosts the basilar membrane (BM) vibrations, which are attenuated when the active process is impaired. However, at high SPLs the contribution of the active process to BM vibration is small compared with the dominating passive mechanical properties of the BM. Consequently, reduction of active cochlear amplification will have greatest effect on BM vibrations and DPOAEs at low SPLs. To distinguish between the "two sources" and the "single source" hypotheses we analyzed the level dependence of the notch and corresponding phase discontinuity in plots of DPOAE magnitude and phase as functions of the level of the primaries. In experiments where furosemide was used to reduce cochlear amplification, an upward shift of the notch supports the conclusion that both the low- and high-level DPOAEs are generated by a single source, namely a nonlinear amplifier with saturating I/O characteristic. PMID- 12083211 TI - Asymmetry of masking between complex tones and noise: the role of temporal structure and peripheral compression. AB - Thresholds for the detection of harmonic complex tones in noise were measured as a function of masker level. The rms level of the masker ranged from 40 to 70 dB SPL in 10-dB steps. The tones had a fundamental frequency (F0) of 62.5 or 250 Hz, and components were added in either cosine or random phase. The complex tones and the noise were bandpass filtered into the same frequency region, from the tenth harmonic up to 5 kHz. In a different condition, the roles of masker and signal were reversed, keeping all other parameters the same; subjects had to detect the noise in the presence of a harmonic tone masker. In both conditions, the masker was either gated synchronously with the 700-ms signal, or it started 400 ms before and stopped 200 ms after the signal. The results showed a large asymmetry in the effectiveness of masking between the tones and noise. Even though signal and masker had the same bandwidth, the noise was a more effective masker than the complex tone. The degree of asymmetry depended on F0, component phase, and the level of the masker. The maximum difference between masked thresholds for tone and noise was about 28 dB; this occurred when the F0 was 62.5 Hz, the components were in cosine phase, and the masker level was 70 dB SPL. In most conditions, the growth-of-masking functions had slopes close to 1 (on a dB versus dB scale). However, for the cosine-phase tone masker with an F0 of 62.5 Hz, a 10-dB increase in masker level led to an increase in masked threshold of the noise of only 3.7 dB, on average. We suggest that the results for this condition are strongly affected by the active mechanism in the cochlea. PMID- 12083213 TI - Mechanisms of modulation gap detection. AB - It has been postulated that the central auditory system contains an array of modulation filters, each responsive to a different range of modulation frequencies present at the outputs of the (peripheral) auditory filters. In the present experiments, we tested what we call the "dip hypothesis," that a gap in modulation is detected using the "dip" in the output of the modulation filter tuned to the modulator frequency. In experiment 1, the task was to detect a gap in the sinusoidal amplitude modulation imposed on a 4-kHz carrier. The modulator preceding the gap ended with a positive-going zero-crossing. There were three conditions, differing in the phase at which the modulator started at the end of the gap; zero-phase, at a positive-going zero-crossing; pi-phase, at a negative going zero-crossing; and "preserved" phase, at the phase the modulator would have had if it had continued without interruption. Modulation frequencies were 5, 10, 20, and 40 Hz. Psychometric functions for detection of the gap were measured using a two-alternative forced-choice task. For the zero-phase and preserved phase conditions, the detectability index, d', increased monotonically with increasing gap duration. For the pi-phase condition, performance was good (d' > 1) for small gap durations, and initially worsened with increasing gap duration, before improving again for longer gap durations. This is the pattern of results expected from the dip hypothesis, provided that the modulation filters have Q values of 2 or more. However, it is also possible that a rhythm cue was used to improve performance in the pi-phase condition for short gap durations; the introduction of the gap markedly disrupted the regular rhythm produced by the modulator peaks. In experiment 2, the rhythm cue was disrupted by varying the modulator period randomly around its nominal value, except for the modulator periods immediately before and after the gap. This markedly impaired performance, and resulted in psychometric functions that were very similar for the zero-phase and pi-phase conditions. This pattern of results is inconsistent with the dip hypothesis. For both experiments, modulation gap "thresholds" (d' approximately 1) were roughly constant when expressed as a proportion of the modulator period. Possible mechanisms of modulation gap detection are discussed and evaluated. PMID- 12083212 TI - Transformation of external-ear spectral cues into perceived delays by the big brown bat, Eptesicus fuscus. AB - The external-ear transfer function for big brown bats (Eptesicus fuscus) contains two prominent notches that vary from 30 to 55 kHz and from 70 to 100 kHz, respectively, as sound-source elevation moves from -40 to +10 degrees. These notches resemble a higher-frequency version of external-ear cues for vertical localization in humans and other mammals. However, they also resemble interference notches created in echoes when reflected sounds overlap at short time separations of 30-50 micros. Psychophysical experiments have shown that bats actually perceive small time separations from interference notches, and here we used the same technique to test whether external-ear notches are recognized as a corresponding time separation, too. The bats' performance reveals the elevation dependence of a time-separation estimate at 25-45 micros in perceived delay. Convergence of target-shape and external-ear cues onto echo spectra creates ambiguity about whether a particular notch relates to the object or to its location, which the bat could resolve by ignoring the presence of notches at external-ear frequencies. Instead, the bat registers the frequencies of notches caused by the external ear along with notches caused by the target's structure and employs spectrogram correlation and transformation (SCAT) to convert them all into a family of delay estimates that includes elevation. PMID- 12083214 TI - Effects of age and frequency disparity on gap discrimination. AB - Temporal discrimination was measured using a gap discrimination paradigm for three groups of listeners with normal hearing: (1) ages 18-30, (2) ages 40-52, and (3) ages 62-74 years. Normal hearing was defined as pure-tone thresholds < or = 25 dB HL from 250 to 6000 Hz and < or = 30 dB HL at 8000 Hz. Silent gaps were placed between 1/4-octave bands of noise centered at one of six frequencies. The noise band markers were paired so that the center frequency of the leading marker was fixed at 2000 Hz, and the center frequency of the trailing marker varied randomly across experimental runs. Gap duration discrimination was significantly poorer for older listeners than for young and middle-aged listeners, and the performance of the young and middle-aged listeners did not differ significantly. Age group differences were more apparent for the more frequency-disparate stimuli (2000-Hz leading marker followed by a 500-Hz trailing marker) than for the fixed frequency stimuli (2000-Hz lead and 2000-Hz trail). The gap duration difference limens of the older listeners increased more rapidly with frequency disparity than those of the other listeners. Because age effects were more apparent for the more frequency-disparate conditions, and gap discrimination was not affected by differences in hearing sensitivity among listeners, it is suggested that gap discrimination depends upon temporal mechanisms that deteriorate with age and stimulus complexity but are unaffected by hearing loss. PMID- 12083215 TI - Efficient adaptive procedures for threshold and concurrent slope estimates for psychophysics and speech intelligibility tests. AB - The minimum standard deviations achievable for concurrent estimates of thresholds and psychometric function slopes as well as the optimal target values for adaptive procedures are calculated as functions of stimulus level and track length on the basis of the binomial theory. The optimum pair of targets for a concurrent estimate is found at the correct response probabilities p1 = 0.19 and p2 = 0.81 for the logistic psychometric function. An adaptive procedure that converges at these optimal targets is introduced and tested with Monte Carlo simulations. The efficiency increases rapidly when each subject's response consists of more than one statistically independent Bernoulli trial. Sentence intelligibility tests provide more than one Bernoulli trial per sentence when each word is scored separately. The number of within-sentence trials can be quantified by the j factor [Boothroyd and Nittrouer, J. Acoust. Soc. Am. 84, 101 114 (1988)]. The adaptive procedure was evaluated with 10 normal-hearing and 11 hearing-impaired listeners using two German sentence tests that differ in j factors. The expected advantage of the sentence test with the higher j factor was not observed, possibly due to training effects. Hence, the number of sentences required for a reliable speech reception threshold (approximately 1 dB standard deviation) concurrently with a slope estimate (approximately 20%-30% relative standard deviation) is at least N = 30 if word scoring for short, meaningful sentences (j approximately 2) is performed. PMID- 12083216 TI - Control of oral closure in lingual stop consonant production. AB - Previous work has shown that the lips are moving at a high velocity when the oral closure occurs for bilabial stop consonants, resulting in tissue compression and mechanical interactions between the lips. The present experiment recorded tongue movements in four subjects during the production of velar and alveolar stop consonants to examine kinematic events before, during, and after the stop closure. The results show that, similar to the lips, the tongue is often moving at a high velocity at the onset of closure. The tongue movements were more complex, with both horizontal and vertical components. Movement velocity at closure and release were influenced by both the preceding and the following vowel. During the period of oral closure, the tongue moved through a trajectory of usually less than 1 cm; again, the magnitude of the movement was context dependent. Overall, the tongue moved in forward-backward curved paths. The results are compatible with the idea that the tongue is free to move during the closure as long as an airtight seal is maintained. A new interpretation of the curved movement paths of the tongue in speech is also proposed. This interpretation is based on the principle of cost minimization that has been successfully applied in the study of hand movements in reaching. PMID- 12083217 TI - An EMA study of VCV coarticulatory direction. AB - This study addresses three issues that are relevant to coarticulation theory in speech production: whether the degree of articulatory constraint model (DAC model) accounts for patterns of the directionality of tongue dorsum coarticulatory influences; the extent to which those patterns in tongue dorsum coarticulatory direction are similar to those for the tongue tip; and whether speech motor control and phonemic planning use a fixed or a context-dependent temporal window. Tongue dorsum and tongue tip movement data on vowel-to-vowel coarticulation are reported for Catalan VCV sequences with vowels /i/, /a/, and /u/, and consonants /p/, /n/, dark /l/, /s/, /S/, alveolopalatal /n/ and /k/. Electromidsagittal articulometry recordings were carried out for three speakers using the Carstens articulograph. Trajectory data are presented for the vertical dimension for the tongue dorsum, and for the horizontal dimension for tongue dorsum and tip. In agreement with predictions of the DAC model, results show that directionality patterns of tongue dorsum coarticulation can be accounted for to a large extent based on the articulatory requirements on consonantal production. While dorsals exhibit analogous trends in coarticulatory direction for all articulators and articulatory dimensions, this is mostly so for the tongue dorsum and tip along the horizontal dimension in the case of lingual fricatives and apicolaminal consonants. This finding results from different articulatory strategies: while dorsal consonants are implemented through homogeneous tongue body activation, the tongue tip and tongue dorsum act more independently for more anterior consonantal productions. Discontinuous coarticulatory effects reported in the present investigation suggest that phonemic planning is adaptative rather than context independent. PMID- 12083218 TI - Timing interference to speech in altered listening conditions. AB - A theory is outlined that explains the disruption that occurs when auditory feedback is altered. The key part of the theory is that the number of, and relationship between, inputs to a timekeeper, operative during speech control, affects speech performance. The effects of alteration to auditory feedback depend on the extra input provided to the timekeeper. Different disruption is predicted for auditory feedback that is out of synchrony with other speech activity (e.g., delayed auditory feedback, DAF) compared with synchronous forms of altered feedback (e.g., frequency shifted feedback, FSF). Stimulus manipulations that can be made synchronously with speech are predicted to cause equivalent disruption to the synchronous form of altered feedback. Three experiments are reported. In all of them, subjects repeated a syllable at a fixed rate (Wing and Kristofferson, 1973). Overall timing variance was decomposed into the variance of a timekeeper (Cv) and the variance of a motor process (Mv). Experiment 1 validated Wing and Kristofferson's method for estimating Cv in a speech task by showing that only this variance component increased when subjects repeated syllables at different rates. Experiment 2 showed DAF increased Cv compared with when no altered sound occurred (experiment 1) and compared with FSF. In experiment 3, sections of the subject's output sequence were increased in amplitude. Subjects just heard this sound in one condition and made a duration decision about it in a second condition. When no response was made, results were like those with FSF. When a response was made, Cv increased at longer repetition periods. The findings that the principal effect of DAF, a duration decision and repetition period is on Cv whereas synchronous alterations that do not require a decision (amplitude increased sections where no response was made and FSF) do not affect Cv, support the hypothesis that the timekeeping process is affected by synchronized and asynchronized inputs in different ways. PMID- 12083219 TI - Anchoring effects in audiovisual speech perception. AB - Two experiments were conducted to investigate whether or not anchoring and selective adaptation induce basically the same psychological effects. The purpose of the first experiment is to show how an audiovisual anchor modifies the perception of consonant-vowel (CV) syllables. The anchors were two purely acoustical, two purely optical, and three audiovisual CV syllables. The results were compared with those of audiovisual speech selective-adaptation experiments conducted by Roberts and Summerfield [Percept. Psychophys. 30, 309-314 (1981)] and Saldana and Rosenblum [J. Acoust. Soc. Am. 95, 3658-3661 (1994)]. The audiovisual anchoring effects were found to be very similar to the audiovisual selective-adaptation effects, but the incompatible audiovisual anchor produced more auditory-based contrast than the purely acoustical anchor or the compatible audiovisual anchor. This difference in contrast had not been found in the previous selective-adaptation experiments. The second experiment was conducted to directly compare audiovisual anchoring and selective-adaptation effects under the same stimuli and with the same subjects. It was found that the compatible audiovisual syllable (AbVb) caused more contrast in selective adaptation than in anchoring, although the discrepant audiovisual syllable (AbVg) caused no difference between anchoring and selective adaptation. It was also found that the anchor AbVg caused more auditory-based contrast than the anchor AbVb. It is suggested that the mechanisms behind these results are different. PMID- 12083220 TI - Quantitative assessment of second language learners' fluency: comparisons between read and spontaneous speech. AB - This paper describes two experiments aimed at exploring the relationship between objective properties of speech and perceived fluency in read and spontaneous speech. The aim is to determine whether such quantitative measures can be used to develop objective fluency tests. Fragments of read speech (Experiment 1) of 60 non-native speakers of Dutch and of spontaneous speech (Experiment 2) of another group of 57 non-native speakers of Dutch were scored for fluency by human raters and were analyzed by means of a continuous speech recognizer to calculate a number of objective measures of speech quality known to be related to perceived fluency. The results show that the objective measures investigated in this study can be employed to predict fluency ratings, but the predictive power of such measures is stronger for read speech than for spontaneous speech. Moreover, the adequacy of the variables to be employed appears to be dependent on the specific type of speech material investigated and the specific task performed by the speaker. PMID- 12083221 TI - Evaluation of context effects in sentence recognition. AB - It was investigated whether the model for context effects, developed earlier by Bronkhorst et al. [J. Acoust. Soc. Am. 93, 499-509 (1993)], can be applied to results of sentence tests, used for the evaluation of speech recognition. Data for two German sentence tests, that differed with respect to their semantic content, were analyzed. They had been obtained from normal-hearing listeners using adaptive paradigms in which the signal-to-noise ratio was varied. It appeared that the model can accurately reproduce the complete pattern of scores as a function of signal-to-noise ratio: both sentence recognition scores and proportions of incomplete responses. In addition, it is shown that the model can provide a better account of the relationship between average word recognition probability (p(e)) and sentence recognition probability (p(w)) than the relationship p(w) =p(e)j, which has been used in previous studies. Analysis of the relationship between j and the model parameters shows that j is, nevertheless, a very useful parameter, especially when it is combined with the parameter j', which can be derived using the equivalent relationship p(w,0) = (1 p(e))(j'), where p(w,0) is the probability of recognizing none of the words in the sentence. These parameters not only provide complementary information on context effects present in the speech material, but they also can be used to estimate the model parameters. Because the model can be applied to both speech and printed text, an experiment was conducted in which part of the sentences was presented orthographically with 1-3 missing words. The results revealed a large difference between the values of the model parameters for the two presentation modes. This is probably due to the fact that, with speech, subjects can reduce the number of alternatives for a certain word using partial information that they have perceived (i.e., not only using the sentence context). A method for mapping model parameters from one mode to the other is suggested, but the validity of this approach has to be confirmed with additional data. PMID- 12083222 TI - The limits of training Japanese listeners to identify English /r/ and /l/: eight case studies. AB - Eight monolingual Japanese listeners were trained to identify English /r/ and /l/ by using 560 training tokens produced by ten talkers in three positions (200 word initial, 200 consonant cluster, and 160 intervocalic tokens). Their baseline performance and transfer of learning were measured using 200 word initial and 200 consonant cluster tokens produced by additional ten talkers. Long-term training (15 days) with feedback indeed increased sensitivity to the nontraining tokens, but tremendous individual differences were found in terms of initial and final sensitivity and response bias. Even after training, however, there remained some tokens for each subject that were misidentified at a level significantly below chance, suggesting that truly nativelike identification of /r/ and /l/ may never be achieved by adult Japanese learners of English. PMID- 12083223 TI - Benefit of modulated maskers for speech recognition by younger and older adults with normal hearing. AB - To assess age-related differences in benefit from masker modulation, younger and older adults with normal hearing but not identical audiograms listened to nonsense syllables in each of two maskers: (1) a steady-state noise shaped to match the long-term spectrum of the speech, and (2) this same noise modulated by a 10-Hz square wave, resulting in an interrupted noise. An additional low-level broadband noise was always present which was shaped to produce equivalent masked thresholds for all subjects. This minimized differences in speech audibility due to differences in quiet thresholds among subjects. An additional goal was to determine if age-related differences in benefit from modulation could be explained by differences in thresholds measured in simultaneous and forward maskers. Accordingly, thresholds for 350-ms pure tones were measured in quiet and in each masker; thresholds for 20-ms signals in forward and simultaneous masking were also measured at selected signal frequencies. To determine if benefit from modulated maskers varied with masker spectrum and to provide a comparison with previous studies, a subgroup of younger subjects also listened in steady-state and interrupted noise that was not spectrally shaped. Articulation index (AI) values were computed and speech-recognition scores were predicted for steady state and interrupted noise; predicted benefit from modulation was also determined. Masked thresholds of older subjects were slightly higher than those of younger subjects; larger age-related threshold differences were observed for short-duration than for long-duration signals. In steady-state noise, speech recognition for older subjects was poorer than for younger subjects, which was partially attributable to older subjects' slightly higher thresholds in these maskers. In interrupted noise, although predicted benefit was larger for older than younger subjects, scores improved more for younger than for older subjects, particularly at the higher noise level. This may be related to age-related increases in thresholds in steady-state noise and in forward masking, especially at higher frequencies. Benefit of interrupted maskers was larger for unshaped than for speech-shaped noise, consistent with AI predictions. PMID- 12083224 TI - Nonlinear analysis of irregular animal vocalizations. AB - Animal vocalizations range from almost periodic vocal-fold vibration to completely atonal turbulent noise. Between these two extremes, a variety of nonlinear dynamics such as limit cycles, subharmonics, biphonation, and chaotic episodes have been recently observed. These observations imply possible functional roles of nonlinear dynamics in animal acoustic communication. Nonlinear dynamics may also provide insight into the degree to which detailed features of vocalizations are under close neural control, as opposed to more directly reflecting biomechanical properties of the vibrating vocal folds themselves. So far, nonlinear dynamical structures of animal voices have been mainly studied with spectrograms. In this study, the deterministic versus stochastic (DVS) prediction technique was used to quantify the amount of nonlinearity in three animal vocalizations: macaque screams, piglet screams, and dog barks. Results showed that in vocalizations with pronounced harmonic components (adult macaque screams, certain piglet screams, and dog barks), deterministic nonlinear prediction was clearly more powerful than stochastic linear prediction. The difference, termed low-dimensional nonlinearity measure (LNM), indicates the presence of a low-dimensional attractor. In highly irregular signals such as juvenile macaque screams, piglet screams, and some dog barks, the detectable amount of nonlinearity was comparatively small. Analyzing 120 samples of dog barks, it was further shown that the harmonic-to-noise ratio (HNR) was positively correlated with LNM. It is concluded that nonlinear analysis is primarily useful in animal vocalizations with strong harmonic components (including subharmonics and biphonation) or low-dimensional chaos. PMID- 12083225 TI - Acoustic identification of female Steller sea lions (Eumetopias jubatus). AB - Steller sea lion (Eumetopias jubatus) mothers and pups establish and maintain contact with individually distinctive vocalizations. Our objective was to develop a robust neural network to classify females based on their mother-pup contact calls. We catalogued 573 contact calls from 25 females in 1998 and 1323 calls from 46 females in 1999. From this database, a subset of 26 females with sufficient samples of calls was selected for further study. Each female was identified visually by marking patterns, which provided the verification for acoustic identification. Average logarithmic spectra were extracted for each call, and standardized training and generalization datasets created for the neural network classifier. A family of backpropagation networks was generated to assess relative contribution of spectral input bandwidth, frequency resolution, and network architectural variables to classification accuracy. The network with best overall generalization accuracy (71%) used an input representation of 0-3 kHz of bandwidth at 10.77 Hz/bin frequency resolution, and a 2:1 hidden:output layer neural ratio. The network was analyzed to reveal which portions of the call spectra were most influential for identification of each female. Acoustical identification of distinctive female acoustic signatures has several potentially important conservation applications for this endangered species, such as rapid survey of females present on a rookery. PMID- 12083226 TI - Temporary shift in masked hearing thresholds in odontocetes after exposure to single underwater impulses from a seismic watergun. AB - A behavioral response paradigm was used to measure masked underwater hearing thresholds in a bottlenose dolphin (Tursiops truncatus) and a white whale (Delphinapterus leucas) before and after exposure to single underwater impulsive sounds produced from a seismic watergun. Pre- and postexposure thresholds were compared to determine if a temporary shift in masked hearing thresholds (MTTS), defined as a 6-dB or larger increase in postexposure thresholds, occurred. Hearing thresholds were measured at 0.4, 4, and 30 kHz. MTTSs of 7 and 6 dB were observed in the white whale at 0.4 and 30 kHz, respectively, approximately 2 min following exposure to single impulses with peak pressures of 160 kPa, peak-to peak pressures of 226 dB re 1 microPa, and total energy fluxes of 186 dB re 1 microPa2 x s. Thresholds returned to within 2 dB of the preexposure value approximately 4 min after exposure. No MTTS was observed in the dolphin at the highest exposure conditions: 207 kPa peak pressure, 228 dB re 1 microPa peak-to peak pressure, and 188 dB re 1 microPa2 x s total energy flux. PMID- 12083227 TI - Should surgeons be tested for blood-borne pathogens? PMID- 12083228 TI - Protecting patients from surgical hepatitis C virus infection. PMID- 12083229 TI - Safer generation of spring-loaded fingerstick lancets. PMID- 12083230 TI - Incidence of nosocomial infection in a brand-new hospital. PMID- 12083231 TI - Preventing transmission of hepatitis B virus from surgeons to patients. PMID- 12083232 TI - More should be done to protect surgical patients from intraoperative hepatitis B infection. PMID- 12083233 TI - Transmission of hepatitis B virus from a surgeon to his patients during high-risk and low-risk surgical procedures during 4 years. AB - OBJECTIVE: We investigated cases of acute hepatitis B in The Netherlands that were linked to the same general surgeon who was infected with hepatitis B virus (HBV). DESIGN: A retrospective cohort study was conducted of 1,564 patients operated on by the surgeon. Patients were tested for serologic HBV markers. A case-control study was performed to identify risk factors. RESULTS: The surgeon tested positive for hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) with a high viral load. He was a known nonresponder after HBV vaccination and had apparently been infected for more than 10 years. Forty-nine patients (3.1%) were positive for HBV markers. Transmission of HBV from the surgeon was confirmed in 8 patients, probable in 2, and possible in 18. In the remaining 21 patients, the surgeon was not implicated. Two patients had a chronic HBV infection. One case of secondary transmission from a patient to his wife was identified. HBV DNA sequences from the surgeon were completely identical to sequences from 7 of the 28 patients and from the case of secondary transmission. The duration of the operation and the occurrence of complications during or after surgery were identified as independent risk factors. Although the risk of HBV infection during high-risk procedures was 7 times higher than that during low risk procedures, at least 8 (28.6%) of the 28 patients were infected during low risk procedures. CONCLUSIONS: Transmission of HBV from surgeons to patients at a low rate can remain unnoticed for a long period of time. Prevention requires a more stringent strategy for vaccination and testing of surgeons and optimization of infectious disease surveillance. Policies allowing HBV-infected surgeons to perform presumably low-risk procedures should be reconsidered. PMID- 12083234 TI - Nosocomial transmission of hepatitis B virus infection among residents with diabetes in a skilled nursing facility. AB - OBJECTIVE: To identify exposures associated with acute hepatitis B virus (HBV) infection among residents with diabetes in a skilled nursing facility. DESIGN: Residents from Unit 3 and other skilled nursing facility residents with diabetes were tested for serologic evidence of HBV infection. Two retrospective cohort studies were conducted. Potential routes of HBV transmission were evaluated by statistical comparison of attack rates. SETTING: A 269-bed skilled nursing facility. PARTICIPANTS: All skilled nursing facility residents with diabetes and skilled nursing facility residents who lived on the same unit as the index case (Unit 3) for some time during the case's incubation period. RESULTS: All 5 residents with acute HBV infection had diabetes and resided in Unit 3. The attack rate among the 12 patients with diabetes in Unit 3 was 42%, compared with 0% among 43 patients without diabetes (relative risk, 37.2; 95% confidence interval, 4.7 to infinity). Acutely infected patients with diabetes received more morning insulin doses (P = .05), and more insulin doses (P = .03) and finger sticks (P = .02) on Wednesdays than did noninfected patients with diabetes. Two chronically infected patients with diabetes in Unit 3 were positive for hepatitis B e antigen and regularly received daily insulin and finger sticks. Of the 4 acute and 3 chronically infected residents from whom HBV DNA was amplified, all were genotype F and had an identical 678-bp S region sequence. Although no component of the lancets or injection devices was shared among residents, opportunities for HBV contamination of diabetes care supplies were identified. CONCLUSIONS: Contamination of diabetes care supplies resulted in resident-to-resident transmission of HBV. In any setting in which diabetes care is performed, staff need to be educated regarding appropriate infection control practices. PMID- 12083235 TI - High prevalence of hepatitis C infection among patients receiving hemodialysis at an urban dialysis center. AB - OBJECTIVE: To determine the seroprevalence and risk factors for hepatitis C virus (HCV) infection among patients at an urban outpatient hemodialysis center. METHODS: This was a cross-sectional study of 227 patients undergoing hemodialysis at the Rogosin Kidney Center on December 15, 1998, with a response rate of 90% (227 of 253). Laboratory records were used to retrieve the total number of blood transfusions received and serologic study results. Univariate and multivariate analyses were used to examine the relationship among HCV serostatus, patient demographics, and HCV risk factors (eg, intravenous drug use [IVDU], intranasal cocaine use, multiple sexual partners, comorbidities, length of time receiving hemodialysis, and total number of blood transfusions received). RESULTS: The seroprevalence of antibody to HCV (anti-HCV) was 23.3% (53 of 227) in the population. In univariate analysis, factors associated with HCV seropositivity included male gender, younger age, history of IVDU, history of intranasal cocaine use, history of multiple sexual partners, human immunodeficiency virus coinfection, increased time receiving dialysis, history of renal transplant, and positive antibody to hepatitis B core antigen. Multivariate logistic regression analysis showed that longer duration receiving dialysis and a history of IVDU were the only risk factors that remained independently associated with HCV seropositivity CONCLUSIONS: HCV is markedly more common in our urban cohort of patients receiving hemodialysis compared with patients receiving dialysis nationally and is associated with a longer duration of receiving dialysis and a history of IVDU. Stricter and more frequent enforcement of universal precautions may be required in hemodialysis centers located in areas with a high prevalence of HCV infection or IVDU among the general population. PMID- 12083236 TI - Occupational risk of blood-borne viruses in healthcare workers: a 5-year surveillance program. AB - OBJECTIVE: This study presents the results of a 5-year surveillance program involving the prospective follow-up of healthcare workers (HCWs) in the Veneto region of Italy exposed to blood-borne viruses. DESIGN: All HCWs who reported an occupational exposure to blood-borne infection joined the surveillance program. Both HCWs and patients were tested for viral markers (hepatitis B surface antigen [HBsAg], antibody to hepatitis B surface antigen [anti-HBs], antibody to hepatitis B core antigen [anti-HBc], antibody to hepatitis C virus [anti-HCV], HCV RNA, and antibody to human immunodeficiency virus [HIV]) and had these markers plus transaminases assayed at 3, 6, and 12 months and then yearly thereafter. Moreover, a program of hepatitis B virus (HBV) prophylaxis was offered to those whose anti-HBs levels were less than 10 IU/mL. PARTICIPANTS: Two hundred forty-five HCWs (156 women and 89 men) with a mean age of 37 (+/- 10) years who reported occupational exposure during the 5-year period. RESULTS: At the time of exposure, 1 HCW was positive for HBsAg (0.4%) and 2 were positive for HCV RNA (0.8%). Among the patients involved, 28 (11.4%) were positive for HBsAg, 68 (27.8%) were positive for HCV RNA, 6 (2.4%) were positive for HIV, and 147 (60.0%) were negative for all viral markers (4 patients were positive for both HCV and HIV). During the follow-up period after exposure (mean, 2.7 [+/- 1.6] years), there was no increase in transaminases or seroconversions to any of the viral markers. CONCLUSION: Our accurate postexposure follow-up revealed a lack of transmission of HBV, HCV, and HIV. PMID- 12083237 TI - Outbreak of hepatitis C virus infection in a hemodialysis unit: potential transmission by the hemodialysis machine? AB - OBJECTIVE: To identify the routes of transmission during an outbreak of infection with hepatitis C virus (HCV) genotype 2a/2c in a hemodialysis unit. DESIGN: A matched case-control study was conducted to identify risk factors for HCV seroconversion. Direct observation and staff interviews were conducted to assess infection control practices. Molecular methods were used in a comparison of HCV infecting isolates from the case-patients and from patients infected with the 2a/2c genotype before admission to the unit. SETTING: A hemodialysis unit treating an average of 90 patients. PATIENTS: A case-patient was defined as a patient receiving hemodialysis with a seroconversion for HCV genotype 2a/2c between January 1994 and July 1997 who had received dialysis in the unit during the 3 months before the onset of disease. For each case-patient, 3 control patients were randomly selected among all susceptible patients treated in the unit during the presumed contamination period of the case-patient. RESULTS: HCV seroconversion was associated with the number of hemodialysis sessions undergone on a machine shared with (odds ratio [OR] per additional session, 1.3; 95% confidence interval [CI95], 0.9 to 1.8) or in the same room as (OR per additional session, 1.1; CI95, 1.0 to 1.2) a patient who was anti-HCV (genotype 2a/2c) positive. We observed several breaches in infection control procedures. Wetting of transducer protectors in the external pressure tubing sets with patient blood reflux was observed, leading to a potential contamination by blood of the pressure-sensing port of the machine, which is not accessible to routine disinfection. The molecular analysis of HCV infecting isolates identified among the case-patients revealed two groups of identical isolates similar to those of two patients infected before admission to the unit. CONCLUSIONS: The results suggest patient-to-patient transmission of HCV by breaches in infection control practices and possible contamination of the machine. No additional cases have occurred since the reinforcement of infection control procedures and the use of a second transducer protector. PMID- 12083238 TI - Low rate of false-positive results with use of a rapid HIV test. AB - BACKGROUND: Occupational exposure to human immunodeficiency virus (HIV) is an important threat to healthcare workers. Centers for Disease Control and Prevention guidelines recommend prompt institution of prophylaxis. This requires (1) immediate prophylaxis after exposure, pending test results that may take more than 24 hours in many hospitals; or (2) performance of a rapid test. The Single Use Diagnostic System (SUDS) HIV-1 Test is used to screen rapidly for antibodies to HIV type 1 in plasma or serum, with a reported sensitivity of more than 99.9%. We used this test from January 1999 until September 2000, when it was withdrawn from the market following reports claiming a high rate of false-positive results. METHODS: We reviewed the results of postexposure HIV testing during 21 months. RESULTS: A total of 884 SUDS tests were performed on source patients after occupational exposures (883 negative results, 1 reactive result). The results of repeat SUDS testing on the reactive specimen were also reactive, but the results of enzyme immunoassay and Western blot testing were negative. A new specimen from the same patient showed a negative result on SUDS testing. This suggested a specificity of 99.9%. In the 4 months after SUDS testing was suspended, there was 1 false-positive result on enzyme immunoassay for 1 of 132 source patients (presumed specificity, 99.2%). CONCLUSION: Use of the SUDS test facilitated rapid and accurate evaluation of source specimens, obviating unnecessary prophylaxis. PMID- 12083239 TI - Patient-to-patient transmission of nosocomial malaria in Italy. AB - OBJECTIVE: To describe nosocomial transmission of malaria from patient to patient via blood exposure. PATIENTS: A 56-year-old man was admitted to an Italian hospital with fever and Plasmodium falciparum parasitemia, but with no risk factors for malaria. Twenty days earlier, he had been admitted for bronchopulmonary disease to the hospital's intensive care unit, where a woman with P. falciparum malaria acquired abroad was present. METHODS: We reviewed both patients' medical records and searched for mosquitoes in the hospital and on the grounds. We interviewed the staff about patient care practices potentially involving contact with blood. The genetic identities of strains were determined by genotyping of the DNA extracted from blood. RESULTS: Molecular genotyping showed that the two strains were identical. The only invasive procedures performed on both patients by the same staff on the same shift were capillary blood sampling by finger stick, intravenous drug administration, and substitution of total parenteral nutrition bags and intravenous sets. The fingerstick device used was designed to prevent person-to-person transmission of blood-borne infections, and the staff interviews did not reveal any incorrect use of aseptic techniques. The likely source of infection was identified during a training course 6 months later: a nurse reported that, when collecting blood, she placed patients' fingers directly on the blood glucose meter, a practice she had learned from a poster advertising the device. CONCLUSIONS: A nosocomial case of malaria was ascertained, which was likely due to patient-to-patient transmission via a contaminated blood glucose meter. Incomplete instructions for the meter seem to have played a role in this case. PMID- 12083241 TI - A nosocomial and occupational cluster of hepatitis A virus infection in a pediatric ward. AB - We describe a cluster of acute hepatitis A virus (HAV) infection that involved two patients and one physician in the pediatric unit where two children with acute HAV infection had been housed. An interview with the unit personnel revealed several breaches in infection control measures and the lack of vaccination of healthcare workers against HAV . PMID- 12083240 TI - Risk of hepatitis C after immunoadsorption. AB - An episode of acute hepatitis in a patient with hemophilia during immunoadsorption therapy initially was misinterpreted as a reactivated hepatitis C virus (HCV) infection, but ultimately was shown to be an exogenous reinfection during cohort treatment with another HCV-positive patient. This incident illustrates that policies for the prevention of nosocomial transmission of blood borne pathogens, especially in cohort treatment units, may need to be reassessed. PMID- 12083242 TI - Transmission of drug-resistant HIV after an occupational exposure despite postexposure prophylaxis with a combination drug regimen. AB - We documented a case of occupational human immunodeficiency virus (HIV) despite postexposure prophylaxis (PEP) with a combination drug regimen after percutaneous injury with a needle from a sharps disposal container in the hospital room of an HIV-infected patient. This failure of PEP with a combination drug regimen may have been related to antiretroviral drug resistance, other factors, or both. This case highlights the importance of preventing injury to prevent occupational transmission of HIV. PMID- 12083243 TI - Determination of tiamulin in type C medicated swine feeds using high throughput extraction with liquid chromatography. AB - An improved method for extraction and analysis of tiamulin is presented to address issues that arose during routine analysis of Type C medicated swine feeds under the current U.S. Food and Drug Administration-Center for Veterinary Medicine (FDA-CVM) approved method. The issues included the need for higher sample throughput and the ability to accommodate a wider variety of feed matrixes. Changes to the FDA-CVM approved method include reduced sample size and solvent volumes, phosphate buffering of tartaric acid, centrifugation, and use of a new liquid chromatography column and adjusted mobile phase composition. A paired sample study was performed to compare performance of the new and existing methods. The paired sample study showed no statistical difference between sample means of paired sets of 17 samples analyzed by both methods (t = 1.95 at 0.05 significance level, p = 0.068). A recovery study showed the method precision to be 2.06% (coefficient of variation) with an average standard recoveryof 95.8%. Ruggedness test results indicated good overall ruggedness of the method. PMID- 12083244 TI - Analysis of grass silage from Northwestern Spain by near-infrared reflectance spectroscopy. AB - Near-infrared reflectance spectroscopy (NIRS) was evaluated for the determination of protein, crude fiber (CF), acid detergent fiber (ADF), and neutral detergent fiber (NDF) in grass silage. Calibration equations were based on analyses of 366 samples of grass silage produced in Northwestern Spain over 4 consecutive years (1992-1995) and validated by analyses of a set of 72 silage samples harvested during 1996. Dried and ground samples were analyzed by chemical and NIRS procedures. The spectral data were analyzed by regression against a range of chemical parameters, using modified partial least-squares (MPLS) multivariate analysis in conjunction with different mathematical treatments of the spectra. For each parameter, the optimum calibration was evaluated on the basis of the coefficient of multiple determination (R2), the coefficient of simple correlation (r2), the standard error of calibration (SEC), the standard error of cross validation (SECV), and the standard error of validation (SEV). R2 and r2 were >0.90; SEC values were 0.58, 1.04, 1.40, and 1.75; SECV values were 0.64,1.15,1.50, and 2.04; and SEV values were 0.56,1.02, 1.42, and 1.80 for protein, CF, ADF, and NDF, respectively. The ratio of the standard deviation of the reference data to the SEV was >3.0 for each of the 4 parameters, which indicates that the equations can be used in routine analysis. PMID- 12083246 TI - Determination of WR-1065 and WR-33278 by liquid chromatography with electrochemical detection. AB - A liquid chromatographic method using electrochemical detection is presented for measuring the thiol WR-1065 12-(3-aminopropylamino) ethanethiol] and its symmetrical disulfide WR-33278 [NH2(CH2)3NHCH2CH2S]2. WR- 1065 is the active, radioprotective drug derived from the phosphorothioate pro-drug WR-2721 (amifostine). External standard curves for both compounds were linear over the range of 40-200 pmol injected (r2 = 0.999 and 0.996 for the thiol and disulfide, respectively). The detection and quantitation limits for WR-1065 were 9 and 18 pmol, respectively, whereas the corresponding values for WR-33278 were 30 and 59 pmol, respectively. Within- and between-day determinations of measurement Vision and accuracy for both compounds validated the suitability of this assay method. PMID- 12083245 TI - Comparison of microbial receptor assay and liquid chromatography for determination of penicillin G and amoxicillin in milk powder. AB - A microbial receptor assay (Charm II Tablet Beta-Lactam Test) and liquid chromatography (LC) were compared for determination of penicillin G (PG) and amoxicillin (AMOX) in reconstituted milk powder. Nonfat dry milk and whole dry milk were reconstituted (10%, w/v) to concentrations of 0, 2.5, 5, 7.5, and 10 ppb PG; nonfat dry milk was reconstituted (10%, w/v) to 0, 7.5, 10, and 15 ppb AMOX. Reconstituted samples were analyzed blindly by each method. Concentrations determined by both methods demonstrated good agreement. A significant difference between methods (p < or = 0.05) was observed only for 7.5 ppb PG in defatted dry milk. Significant differences were not observed between known concentrations and concentrations determined by the Charm II assay for PG or AMOX in defatted dry milk and PG in whole dry milk. Results by LC showed significant differences (p < 0.05) between known and measured concentrations at 10 ppb PG in both milks and 0 ppb AMOX in defatted dry milk. These results suggest that both the microbial receptor assay and LC may be useful for determination of PG and AMOX near safe level and tolerance, respectively, in reconstituted milk powder. PMID- 12083247 TI - Simultaneous determination of promethazine, chlorpromazine, and perphenazine by multivariate calibration methods and derivative spectrophotometry. AB - Partial least-squares (PLS) regression, singular value decomposition-based PLS, and an artificial neural network (ANN) were tested as calibration procedures for the simultaneous determination of promethazine, chlorpromazine, and perphenazine by both conventional and derivative spectrophotometry. Comparison of the results revealed that the application of the ANN to the derivative spectra is superior to the application of the 2 PLS methods used. Different binary and ternary synthetic mixtures of the phenothiazine drugs in pure form and in tablets were analyzed by the proposed method, and acceptable results were obtained. PMID- 12083248 TI - Validation of a gas chromatographic method for determination of fatty alcohols in 10 mg film-coated tablets of policosanol. AB - A gas chromatographic method using a packed column and 1-eicosanol as an internal standard was validated for the determination of the fatty alcohols that compose policosanol in 10 mg film-coated tablets. The alcohols were determined as trimethylsilyl derivatives, prepared with N-methyl-N trimethylsilyltrifluoroacetamide. The method can detect degradation products with high retention times, without interfering with the peaks of the active principle. Good linearity (correlation coefficient = 0.9992) and accuracy (mean recovery 100.27 +/- 1.66%) were proven over a range of 25-200% of the nominal concentration. Within- and between-day precision at the nominal 100% value met the acceptance criteria (<2%). Ruggedness was examined through an intralaboratory experimental study in which 6 operational changes were made; the changes were found to have no effect on quantitation, repeatability, resolution, and relative retention time. The method is suitable for the quality control process and stability studies of these tablets. PMID- 12083249 TI - Determination of chondroitin sulfate in raw materials by liquid chromatography. AB - A rapid method for the determination of chondroitin sulfate in raw materials was developed. The samples were finely powdered, dissolved in water, and injected directly into the liquid chromatograph. The method used a C18 column, a wavelength of 195 nm, and a mobile phase containing octane sulfonic acid. The method gave results that were slightly different from those generated by atitrimetric method using cetylpyridinium chloride. PMID- 12083250 TI - Determination of nalidixic acid by fluorometry with sodium borohydride and hydrogen peroxide. AB - A simple and sensitive fluorometric method for the determination of nalidixic acid was established by using 0.75M sodium borohydride and 7.5% hydrogen peroxide solution as fluorogenic reagents. Analyte concentrations of 0.0232-11.6 pg/mL could be determined with high precision and accuracy by the method. A relative standard deviation of 1.75% was obtained for a nalidixic acid concentration of 0.232 pg/mL. The method was satisfactorily applied to the determination of nalidixic acid in human serum, fish muscle, and chicken muscle, and the calibration curves were linear from 0.23 to 58.00 pg/mL, from 9.28 to 32.48 mg/kg, and from 4.64 to 23.20 mg/kg, respectively. The specificity of the reaction is also discussed. PMID- 12083251 TI - Evaluation of VIDAs Immuno-concentration Salmonella assay Plus selective plate method (Hektoen enteric, bismuth sulfite, Salmonella identification) for detection of Salmonella in selected foods: collaborative study. AB - The VIDAS Immuno-concentration Salmonella (ICS) plus selective plate method (Hektoen enteric, bismuth sulfite, Salmonella identification) method for the detection of Salmonella was compared to the Bacteriological Analytical Manual (BAM)/AOAC culture method in a collaborative study. Thirty-two laboratories participated in the evaluation. Each laboratory tested one or more of the 6 test products: milk chocolate, nonfat dry milk, dried whole egg, soy flour, ground black pepper, and ground raw turkey. The 2 methods were in agreement for 1,283 of the 1,440 test samples. Of the 157 test samples not in agreement, 82 were VIDAS ICS plus selective plate-positive and BAM/AOAC-negative, and 75 were VIDAS ICS plus selective plate-negative and BAM/AOAC-positive. PMID- 12083253 TI - Evaluation of VIDas immuno-concentration Salmonella/VIDAS salmonella immunoassay method for detection of Salmonella in selected foods: collaborative study. AB - The VIDAS Immuno-concentration Salmonella ICS)/VIDAS Salmonella (SLM) immunoassay method for the detection of Salmonella was compared to the Bacteriological Analytical Manual (BAM)/AOAC culture method in a collaborative study. Thirty-two laboratories participated in the evaluation. Each laboratory tested one or more of the 6 test products: milk chocolate, nonfat dry milk, dried whole egg, soy flour, ground black pepper, and ground raw turkey. The 2 methods were in agreement for 1,266 of the 1,440 samples. Of the 174 samples not in agreement, 69 were VIDAS CS/SLM-positive and BAM/AOAC-negative and 105 were VIDAS ICS/SLM negative and BAM/AOAC-positive. PMID- 12083252 TI - Salmonella in selected foods by VIDAS immuno-concentration Salmonella plus selective plate method (Hektoen enteric, xylose lysine desoxycholate, bismuth sulfite): collaborative study. AB - The VIDAS Immuno-concentration Salmonella (ICS) plus selective plate method (Hektoen enteric, xylose lysine desoxycholate, bismuth sulfite) method for the detection of Salmonella was compared to the Bacteriological Analytical Manual (BAM)/AOAC culture method in a collaborative study. Thirty-two laboratories participated in the evaluation. Each laboratory tested one or more of the 6 test products: milk chocolate, nonfat dry milk, dried whole egg, soy flour, ground black pepper, and ground raw turkey. The 2 methods were in agreement for 1,297 of the 1,455 samples. Of the 158 samples not in agreement, 82 were VIDAS ICS plus selective plate-positive and BAM/AOAC-negative, and 76 were VIDAS ICS plus selective plate-negative and BAM/AOAC-positive. PMID- 12083254 TI - Determination of mercury in seafood by flow injection-cold vapor atomic absorption spectrometry after microwave digestion: NMKL interlaboratory study. Nordic Committee on Food Analysis. AB - Ten laboratories participated in an interlaboratory method-performance (collaborative) study of a method for the determination of mercury in foods of marine origin by flow injection-cold vapor atomic absorption spectrometry after wet digestion using a microwave oven technique. The study was preceded by a training round of samples of known identity. The method was tested on a total of 7 seafood products: blue mussel (Mytilus edulis), cod muscle (Gadus morhua), crab (Cancer pagurus), scampi (Nephrops norwegicus), black scabbard fish (Aphnopus carbo), longnose velvet dogfish (Centroscymus crepidater), and Portuguese dogfish (Cenbroscymus coelolepis) with mercury concentrations of 0.14, 0.24, 0.35, 0.59,11.42, 4.2, and 13.2 microg/g, respectively. The materials were presented to the participants in the study as blind duplicates, and the participants were asked to perform single determinations on each sample. Repeatability relative standard deviations (RSDr) for mercury ranged from 2.4 to 14.0%. Reproducibility relative standard deviations (RSDR) ranged from 7.7 to 16.6%. HORRAT values for all samples were <1.0. PMID- 12083255 TI - Assessment of MIST Alert, a commercial qualitative assay for detection of paralytic shellfish poisoning toxins in bivalve molluscs. AB - A recently developed commercial rapid test kit (MIST Alert) was assessed for determination of the presence of paralytic shellfish poisoning (PSP) toxins in shellfish. Several commercially important shellfish species obtained from the UK shellfish toxin monitoring program, containing a range of total PSP toxicities as determined by the mouse bioassay (MBA), were tested. The kit detected toxin in all samples containing the European Community tolerance level of 80 microg saxitoxin (STX) equivalents/100 g shellfish flesh as determined by the MBA. With one exception, the kit detected toxin in all samples that contained >40 microg STX equivalents/100 g according to the MBA. Among samples in which the MBA did not detect toxin, the kit disagreed in 25% of the tests, although further analysis by liquid chromatography (LC) and MBA of some samples confirmed the presence of toxins. These results suggest that MIST Alert may be suitable as an initial screen for PSP toxins as part of routine monitoring programs, thereby greatly reducing the number of MBAs. Trials were also performed by nonscientific personnel to evaluate the ease of use and interpretation of results obtained by MIST Alert. The results indicated that the kits could be readily used and accurately interpreted by individuals with no technical or scientific background. PMID- 12083256 TI - Cleanup procedure for determination of aflatoxins in major agricultural commodities by liquid chromatography. AB - A simple, fast, reliable, and inexpensive chemical cleanup procedure was developed for quantitation of aflatoxins in major important agricultural commodities by liquid chromatography (LC). Aflatoxins were extracted from a ground sample with methanol-water (80 + 20, v/v), and after a single cleanup step on a minicolumn packed with basic aluminum oxide, they were quantitated by LC equipped with a C18 column, photochemical reactor, and fluorescence detector. Water-methanol-1-butanol (1,400 + 720 + 25, v/v/v) served as the mobile phase. Recoveries of aflatoxins B1, B2, G1, and G2 from peanuts spiked at 5.0, 2.5, 7.5, and 2.5 microg/kg were 87.2 +/- 2.3, 82.0 +/- 0.8, 80.0 +/- 1.8, and 80.4 +/- 2.8%, respectively. Similar recoveries, precision, and accuracy were achieved for corn, cottonseed, almonds, Brazil nuts, pistachios, and walnuts. The quantitation limit for aflatoxin B1 was 1 microg/kg. The minimal cost of the minicolumn allows for substantial savings compared with available commercial aflatoxin cleanup devices. PMID- 12083257 TI - Real-time quantitative polymerase chain reaction methods for four genetically modified maize varieties and maize DNA content in food. AB - Quantitative detection methods are needed for enforcement of the recently introduced labeling threshold for genetically modified organisms (GMOs) in food ingredients. This labeling threshold, which is set to 1% in the European Union and Switzerland, must be applied to all approved GMOs. Four different varieties of maize are approved in the European Union: the insect-resistant Bt176 maize (Maximizer), Btl 1 maize, Mon810 (YieldGard) maize, and the herbicide-tolerant T25 (Liberty Link) maize. Because the labeling must be considered individually for each ingredient, a quantitation system for the endogenous maize content is needed in addition to the GMO-specific detection systems. Quantitative real-time polymerase chain reaction detection methods were developed for the 4 approved genetically modified maize varieties and for an endogenous maize (invertase) gene system. PMID- 12083258 TI - Determination of niacin in infant formula by solid-phase extraction/liquid chromatography: peer-verified method performance-interlaboratory validation. AB - This paper reports the results of the interlaboratory peer validation study of AOAC Peer-Verified Method (PVM) 1:2,000 for the determination of niacin in infant formula by solid-phase extraction/liquid chromatography. We have used a Data Quality Objectives (DQO) approach to address not only method variability and robustness but also accuracy of data through the use of an appropriate reference material in conjunction with the interlaboratory validation study. Our DQO included the following: (1) statistical agreement of analytical results and quantitative recovery between 2 collaborating laboratories; (2) the repeatability relative standard deviation (RSDr) values and the HORRAT (Horwitz ratio) obtained (1.07), which satisfied the criteria of the Horwitz "limits of acceptability" at the analyte level present; (3) validation of lack of interference; and (4) accuracy agreement within assigned values for a certified reference material. National Institute of Standards and Technology Standard Reference Material (NIST SRM) 1846 Infant Formula, with a certified value of 63.3 +/- 7.6 microg/g for niacin content, was used as a test material for collaborative study and accuracy assessment. Niacin values obtained by the originating laboratory were 59.7 +/- 4.0 microg/g (95% confidence interval [CI] = 1.4 microg/g with a relative standard deviation [RSD] of 6.7%) and by the peer laboratory were 56.6 +/- 6.6 microg/g (95% CI = 4.1 microg/g, with an RSD of 11.7%). Statistical evaluation using the means equivalence test showed that nicotinic acid values obtained by the peer laboratory were equivalent to those values obtained by the originating laboratory. Linear calibration curves and quantitative recovery were obtained. Integration of the PVM process with a readily available certified reference material gives the user confidence in the accuracy of the data generated by the method through traceability to the reference material used. PMID- 12083259 TI - Measurement of resistant starch. AB - A robust and reliable method was developed to measure resistant starch (RS), i.e., starch that enters the large intestine. In vivo conditions were reflected as much as possible while a user-friendly format was maintained. Parameters investigated included a-amylase concentration, pH of incubation, maltose inhibition of alpha-amylase, the need for amyloglucosidase inclusion, the effect of shaking and stirring on determined values, and problems in recovering and analyzing the RS-containing pellet. The RS values obtained were in good agreement with published in vivo data. An interlaboratory evaluation of the method has been completed (First Action Method 2002.02). PMID- 12083260 TI - Isolation of light filth from ground oregano and ground marjoram: a modification using isopropanol as a defatting agent: in-house study. AB - A procedural modification of the AOAC Official Method for extracting light filth from ground oregano and ground marjoram was tested in an intralaboratory study. The modified method specifies isopropanol defatting, 975.49A(a), rather than chloroform-isopropanol defatting, 975.49A(b), followed by direct flotation as directed in AOAC Official Method, 975.49B(b). The modified method provided comparable results in less time while also providing safety, health, and financial benefits. PMID- 12083261 TI - Determination of vitamin K1 isomers in foods by liquid chromatography with C30 bonded-phase column. AB - Vitamin K1 was determined in a variety of foods by using reversed-phase liquid chromatography with a C30 column followed by post-column reduction to the fluorescent hydroquinone derivatives. Lipids were removed by lipase digestion, followed by single extraction into hydrocarbon, and the protocol was extended to selected natural and processed foods. Biologically active trans- and inactive ci vitamin K1 isomers were measured individually to evaluate the true nutritional status of the products. Method performance parameters confirmed the validity of the technique. The use of the triacontyl-bonded C30 phase for selective phylloquinone isomer measurement extends previously validated AOAC Method 999.15 for vitamin K1 in milk and infant formula to a wider range of foods important in the human diet. The cis-vitamin K1 isomer contributes up to about 15% of total phylloquinone in certain foods. PMID- 12083262 TI - Pesticide residues in agricultural products monitored in Hyogo Prefecture, Japan, FYs 1995-1999. AB - During a 5-year monitoring survey (April 1995-March 2000) of pesticide residues in agricultural products, 765 samples (478 domestic; 287 imported) collected in Hyogo Prefecture, Japan, were analyzed. The number of pesticides tested increased from 107 in fiscal year (FY) 1995 to 204 in FY 1999. The purpose of the study was to promote consumer safety by excluding the food illegally containing pesticide residues from markets. Overall, 51 % of domestic and 32% of imported samples contained no detectable residues. Multiple residues were detected in 152 (32%) of domestic and 146 (51%) of imported samples. The limit of quantitation was set at 0.01 microg/g and the limit of detection was 0.001 microg/g. Most of the residues were present at low concentrations; 70% of detections in domestic samples were <0.05 microg/g, and 97% were <0.5 microg/g. Although 86% of antifungal agent residues in imported citrus fruits were > or = 0.1 microg/g, 59% of the other residues in imported samples were <0.05 microg/g, and 96% were <0.5 microg/g. Violations of maximum residue limits (MRL) were observed in 3 samples: diazinon in chrysanthemums, dieldrin in cucumbers, and bitertanol in bananas. Of the detectable residues above 0.01 microg/g, 55% in domestic and 38% in imported samples were <10% of the MRL. Of all the samples, 2.4% contained more than 5 different pesticides; tomatoes, strawberries, apples, and citrus fruits tended to have more multiple residues. PMID- 12083263 TI - Monitoring of five postharvest fungicides in fruit and vegetables by matrix solid phase dispersion and liquid chromatography/mass spectrometry. AB - A method was developed for monitoring dichloran, flutriafol, o-phenylphenol, prochloraz, and tolclofos-methyl in fruits and vegetables, using matrix solid phase dispersion and liquid chromatography with mass spectrometry detection. The method was used to determine fungicide content in 200 samples of chards, onions, peppers, bananas, lemons, and oranges. Of the samples examined, 54% contained o phenylphenol with concentrations ranging from 0.005 to 3.34 mg/kg and 35% showed prochloraz in the range of 0.06-1.95 mg/kg. Dichloran, flutriafol, and tolclofos methyl were detected only occasionally. Only 4% of the samples exceeded the European Union maximum residue limits. The pesticides involved were tolclofos methyl in 3 samples, o-phenylphenol and flutriafol in 2, and dichloran in one. The calculation of estimated daily intake from these monitoring data showed that dietary intakes were much lower than the acceptable daily intakes established by international agencies. PMID- 12083264 TI - Differential pulse polarographic determination of lead in complex materials after adsorption of its N-methylethylxanthocarbamate complex on microcrystalline naphthalene or on N-methylethylxanthocarbamate-benzyldimethyltetradecylammonium naphthalene adsorbent. AB - Lead is quantitatively adsorbed as the lead N-methylethylxanthocarbamate (MEXC) benzyldimethyltetradecylammonium (BDTA) ion pair complex on microcrystalline naphthalene in the pH range 4.0-11.0. The metal is desorbed with HCI and determined by differential pulse polarography. Alternatively lead can be quantitatively adsorbed on the adsorbent (MEXC-BDTA-naphthalene) packed in a column at a flow rate of 1-2 mL/min and determined similarly. Dissolved oxygen is removed by adding a few milliliters of 4% NaBH4 solution. The detection limit is 0.12 microg/mL at the minimum instrumental settings (signal-to-noise ratio, 2). Linearity was obtained over the concentration range 0.3-20.0 microg/mL with a correlation factor of 0.9998 and a relative standard deviation of +/- 0.98%. Various parameters, such as the effect of pH, volume of aqueous phase, flow rate, and the interference of a large number of metal ions and anions, were studied in detail to optimize the conditions for the trace determination of lead in various standard alloys, standard biological materials, and environmental samples. PMID- 12083265 TI - Preparation of samples for gas chromatography/mass spectrometry analysis of phthalate and adipate esters in plasma and beverages by steam distillation and extraction. AB - Phthalate and adipate esters are present in relatively large amounts in the environment, resulting in their large blank values at analysis and making precise analysis difficult. We developed a highly sensitive analytical method for phthalate and adipate esters in plasma and beverages by lowering the blank values that interfere with analysis. The method uses a closed distillation cleanup system in which steam distillation and extraction are performed simultaneously. The recoveries from beverages and plasma were both satisfactory, ranging from 90.2 to 118.3%, relative standard deviation (RSD) = 2.8-5.3%, and 96.2-134.4%, RSD = 2.2-6.5%, respectively. The detection limits of dibutyl phthalate and di-2 ethyl hexyl phthalate were 5 ng/mL, and those of diethyl phthalate, butyl benzyl phthalate, and di-2-ethyl hexyl adipate were 10 ng/mL in rabbit plasma and beverages. PMID- 12083266 TI - Validation of a method for the determination of multiclass pesticide residues in fruit juices by liquid chromatography/tandem mass spectrometry after extraction by matrix solid-phase dispersion. AB - A multiresidue method was developed and validated for the determination of pesticide residues (omethoate, dimethoate, carbendazim, propoxur, thiabendazole, carbaryl, pirimicarb, azinphos-methyl, methidathion, and iprodione) in fruit juices. The samples were extracted by matrix solid-phase dispersion with diatomaceous earth and analyzed by liquid chromatography/tandem mass spectrometry. The method detection limits were <0.2 ppb for all pesticides; the relative standard deviations for analyses of samples fortified over the range of 2-50 ng/g were <9%, and the recoveries for each pesticide were all between 77 and 102%. The proposed method was used to analyze 21 commercial fruit juices; pesticide residues were found in 71% of the samples. PMID- 12083267 TI - Differential pulse polarographic determination of procymidone in formulations and wine. AB - The dicarboximide fungicide procymidone was studied systematically by using direct current polarography, cyclic voltammetry, differential pulse polarography (DPP), controlled potential electrolysis, and millicoulometry in the universal buffer medium with dimethylformamide as the solvent. Procymidone exhibited a single well-defined polarographic wave in the pH range 2.0-6.0, leading to the formation of the hydroxy compound. The overall reduction process was diffusion controlled and adsorption-free. The variation of half-wave potential with pH, the concentration of the analyte, and other experimental conditions are described. The reduction mechanism proposed is an overall 4-electron process, in which the dicarboximide group is reduced. DPP was used to determine procymidone in agricultural formulations and wine at the optimum conditions found; a detection limit of 2.4 x 10(-9) M was estimated. The results obtained by the proposed method were also compared with those obtained by other methods. PMID- 12083268 TI - Determination of cadmium, aluminium, and copper in beer and products used in its manufacture by electrothermal atomic absorption spectrometry. AB - Procedures were developed for determining cadmium, aluminium, and copper in beer and the products used in its manufacture by electrothermal atomic absorption spectrometry. Beer samples were injected into the furnace and solid samples were introduced as suspensions after preparation in a medium containing hydrogen peroxide, nitric acid, and ammonium dihydrogen phosphate for cadmium atomization. Calibration was performed with aqueous standards, and characteristic masses and detection limits were, respectively, 1 and 0.3 pg for cadmium, 18 and 5.4 pg for aluminium, and 5.6 and 6.8 pg for copper. Different samples of beer, wort, brewer's yeast, malt, raw grain, and hops were analyzed by the proposed procedures. Cadmium was found in low concentrations (0.001-0.08 microg/g and 0 1.3 ng/mL); copper (3-13 microg/g and 25-137 ng/mL) and aluminium (0.6-9 microg/g and 0.1-2 microg/mL) were found at higher levels. The reliability of the procedure was confirmed by comparing the results obtained with others based on microwave oven sample digestion, and by analyzing several certified reference materials. PMID- 12083269 TI - Spectrophotometric determination of caramel content in spirits aged in oak casks. AB - A new methodology was developed for determination of caramel in spirits aged in oak casks. The method is based on differences between the electronic spectra of oak aqueous alcoholic extracts and caramel solutions in the same solvent. The data were treated by 2 different approaches: the simplest one was based on the plot of caramel concentration versus the ratio of absorbance at 210 and 282 nm; the other was based on a partial least squares (PLS) calibration model using the first derivative of the spectral data. Both methodologies were applied to analysis of 159 aged spirit samples. The mean caramel content of several Brazilian sugar cane spirits (cachaca) and all United States whiskies was smaller than that of Scottish whiskies and other brandies from several countries. Correlation was good between caramel concentrations for the same sample calculated by the 2 methods. The uncertainties following PLS and the absorbance ratio method were 0.01 and 0.03 g/L, respectively, for a sample containing 0.45 g/L caramel. Treatment of UV-VIS spectra by pattern recognition using hierarchical clustering analysis and principal components analysis allowed discrimination of the samples as a function of their caramel content. It was possible to distinguish U.S. whiskies from other whiskies, but a clear differentiation among Brazilian cachacas as a function of their geographic origin was not feasible. Small caramel quantities as low as 0.08 g/L were clearly detected by these methodologies. PMID- 12083271 TI - Analytical challenges: bridging the gap from regulation to enforcement. AB - An overview is presented of the analytical steps that may be needed to determine the presence of genetically modified organisms (GMOs) or for analysis of GMO derived produce. The analytical aspects necessary for compliance with labeling regulations are discussed along with bottlenecks that may develop when a plant product or a food sample is analyzed for conformity with current European Union GMO legislation. In addition to sampling and testing, other topics deal with complications that arise from biological and agricultural realities that may influence testing capabilities. The issues presented are intended to serve as elements to examine the different challenges that enforcement laboratories might face. PMID- 12083270 TI - Method development in relation to regulatory requirements for detection of GMOs in the food chain. AB - This is a summary report of a joint workshop held in Brussels, Belgium, in December 2000. The workshop was organized by the ILSI Europe Novel Food Task Force in collaboration with the European Commission's Joint Research Centre (JRC) and ILSI International Food Biotechnology Committee. The purpose was to investigate progress in the development of analytical methods since the last workshop was held in June 1998. PMID- 12083272 TI - Identity preservation of genetically modified organisms in the food chain: requirements, methods, and costs. AB - The use of the technology of genetic modification (GM) in European agriculture and the food supply chain is currently controversial. Because of strong anti-GM technology sentiments, the use of ingredients derived from plants containing GM have largely been eliminated from foods manufactured for direct human consumption by the food supply chain in much of the European Union (EU). During the past year, the attention of those opposed to the technology has turned to the use of GM ingredients in livestock production systems by incorporation of GM soy and maize in animal feed. A discussion is presented of the key issues relating to this subject, focusing on how supplies of GM or non-GM products are segregated or how their identities are preserved. The discussion is centered on GM maize and soybeans into which agronomic traits, such as herbicide tolerance and/or insect resistance, have been incorporated. These are currently the only crops into which some varieties containing GM have been approved for use in the EU. PMID- 12083273 TI - Detection of genetically modified crops and their derivatives: critical steps in sample preparation and extraction. AB - The detection of genetically modified crops in foodstuff relies on detection of transgenic DNA or protein material in the sample matrix. Purified DNA or proteins are used as analytical material for polymerase chain reaction technologies and immunodiagnostics. Successful sample preparation is critical to the validity of subsequent analysis. For routine analysis, a good sample preparation technique should be simple, safe, and inexpensive while reproducibly generating DNA/protein of sufficient quality and yield. The suitability of isolated DNA or protein as an analyte for a detection or characterization technique depends on amount or concentration, purity, and integrity, each of which may be influenced by sample matrix and the extraction technique, and, in turn, may impact the validity of analytical techniques. The key sample preparation steps of homogenization, pretreatment, extraction, and purification are discussed as well as typical analytical methods. Consideration is given to application of these steps for particular sample matrixes to maximize yield, reduce inhibition effects, and minimize contamination. The choice of the most appropriate and valid methods for sample preparation from particular foods is discussed with respect to DNA analysis. Attention is also given to ease of use, cost, and generic applicability of the procedures. PMID- 12083274 TI - Production of certified reference materials for the detection of genetically modified organisms. AB - Certified reference materials (CRMs) are an essenIial tool in the quality assurance of analytical measurements. They are produced, certified, and used in accordance with relevant ISO (International Organization for Standardization) and BCR (Community Bureau of Reference) guidelines. The Institute for Reference Materials and Measurements (IRMM; Geel, Belgium) has produced the first powdery genetically modified organism (GMO) CRMs in cooperation with the Institute for Health and Consumer Protection (Ispra, Italy). Until now, different weight percentages in the range of 0-5% for 4 GMOs in Europe were produced and certified: Bt (Bacillus thuringiensis)-11 and Bt-176 maize, Roundup Ready soybean, and MON810 maize. Bt-11 and Bt-176 maize and Roundup Ready soybean were produced by IRMM on behalf of Fluka Chemie AG (Buchs, Switzerland). Characterization of used base material is the first step in production and is especially important for GMO CRMs. The production of powdery GMO CRMs and methods used for production control are described. Thorough control of homogeneity and stability are essential for certification of reference materials and ensure validity of the certificate for each bottle of a batch throughout a defined shelf life. Because production of reference materials and their maintenance are very labor- and cost-intensive tasks, the usefulness of new types of GMO CRMs must be estimated carefully. PMID- 12083275 TI - Protein immunoassay methods for detection of biotech crops: applications, limitations, and practical considerations. AB - Immunoassay methods are available for detection and quantitation of proteins expressed by most biotechnology-derived crops in commercial production. The 2 most common test formats are enzyme-linked immunosorbent assay (ELISA) and immunochromatographic (lateral flow) strip tests. Two ELISA methods, one for Roundup Ready soybeans and one for MON810 CrylAb corn, were the subject of large international collaborative studies and were demonstrated to quantitatively determine the concentrations of biotech crops in samples of ground grain. Quantitative ELISA methods are also useful for analysis of processed fractions of agricultural commodities such as soybean toasted meal or corn flour. Both strip tests and ELISAs for biotech crops are currently being used on a large scale in the United States to manage the sale and distribution of grain. In these applications, tests are used to determine if the concentration of biotech grain is above or below specified threshold limits. Using existing U.S. Department of Agriculture sampling techniques, the reliability of the threshold determination is expressed in terms of statistical confidence rather than analytical precision. Combining the use of protein immunoassays with Identity Preservation systems provides an effective means of characterizing the raw and processed agricultural inputs to the food production system in a way that allows food producers to comply with labeling laws. PMID- 12083277 TI - State of the art and limitations of quantitative polymerase chain reaction. AB - Consequential to the implementation of European Commission (EC) Regulation 1139/98, EC Regulation 49/2000, and EC Regulation 50/2000 has been the need to measure accurately the levels of the genetically modified (GM) species Roundup Ready Soya and Bt 176 Maize that are present in food. Analytical methods to detect and quantitate these transgenic species have received much attention particularly with respect to the deminimus threshold of 1% for their presence in materials derived from non-GM identity-preserved (IP) supplies. The relative advantages and limitations of threshold analysis by double-competitive polymerase chain reaction (PCR) and quantitative real-time PCR are discussed in their application to the quantitative analysis of processed foods. Consideration is also given to other factors involved in the analyses that affect the performance of quantitative procedures, and to the many uncertainties involved in the precision of a reported analytical result. PMID- 12083276 TI - Detection of genetically modified organisms in foods by protein- and DNA-based techniques: bridging the methods. AB - According to European Commission (EC) Regulation 1139/98, foods and food ingredients that are to be delivered to the final consumer in which either protein or DNA resulting from genetic modification is present, shall be subject to additional specific labeling requirements. Since 1994, genetically altered tomatoes, squash, potatoes, canola, cotton, and soy have been on the market. Recently, insect-resistant and herbicide-tolerant maize varieties have been introduced. Soy and maize are 2 of the most important vegetable crops in the world. During the past 4 years, both protein- and DNA-based methods have been developed and applied for detection of transgenic soy and maize, and their derivatives. For protein-based detection, specific monoclonal and polyclonal antibodies have been developed; for immunochemical detection, Western blot analysis and enzyme-linked immunosorbent assays are the most prominent examples. For detection of genetically modified organisms (GMOs) at the level of DNA, polymerase chain reaction-based methods are mainly used. For these reactions, highly specific primer sets are needed. This study compares the principally different methods. Specificity of methods and the possible risks of false positive or false-negative results are considered in relation to sampling, matrix effects, and food processing procedures. In addition, quantitative aspects of protein- and DNA-based GM detection methods are presented and discussed. This is especially relevant as EC regulation 49/2000, which defines a threshold for an unintentional comingling of 1%, came into force on April 10, 2000. PMID- 12083278 TI - DNA methods: critical review of innovative approaches. AB - The presence of ingredients derived from genetically modified organisms (GMOs) in food products in the market place is subject to a number of European regulations that stipulate which product consisting of or containing GMO-derived ingredients should be labeled as such. In order to maintain these labeling requirements, a variety of different GMO detection methods have been developed to screen for either the presence of DNA or protein derived from (approved) GM varieties. Recent incidents where unapproved GM varieties entered the European market show that more powerful GMO detection and identification methods will be needed to maintain European labeling requirements in an adequate, efficient, and cost effective way. This report discusses the current state-of-the-art as well as future developments in GMO detection. PMID- 12083279 TI - Detection methods and performance criteria for genetically modified organisms. AB - Detection methods for genetically modified organisms (GMOs) are necessary for many applications, from seed purity assessment to compliance of food labeling in several countries. Numerous analytical methods are currently used or under development to support these needs. The currently used methods are bioassays and protein- and DNA-based detection protocols. To avoid discrepancy of results between such largely different methods and, for instance, the potential resulting legal actions, compatibility of the methods is urgently needed. Performance criteria of methods allow evaluation against a common standard. The more-common performance criteria for detection methods are precision, accuracy, sensitivity, and specificity, which together specifically address other terms used to describe the performance of a method, such as applicability, selectivity, calibration, trueness, precision, recovery, operating range, limit of quantitation, limit of detection, and ruggedness. Performance criteria should provide objective tools to accept or reject specific methods, to validate them, to ensure compatibility between validated methods, and be used on a routine basis to reject data outside an acceptable range of variability. When selecting a method of detection, it is also important to consider its applicability, its field of applications, and its limitations, by including factors such as its ability to detect the target analyte in a given matrix, the duration of the analyses, its cost effectiveness, and the necessary sample sizes for testing. Thus, the current GMO detection methods should be evaluated against a common set of performance criteria. PMID- 12083280 TI - Validation studies and proficiency testing. AB - Genetically modified organisms (GMOs) entered the European food market in 1996. Current legislation demands the labeling of food products if they contain <1% GMO, as assessed for each ingredient of the product. To create confidence in the testing methods and to complement enforcement requirements, there is an urgent need for internationally validated methods, which could serve as reference methods. To date, several methods have been submitted to validation trials at an international level; approaches now exist that can be used in different circumstances and for different food matrixes. Moreover, the requirement for the formal validation of methods is clearly accepted; several national and international bodies are active in organizing studies. Further validation studies, especially on the quantitative polymerase chain reaction methods, need to be performed to cover the rising demand for new extraction methods and other background matrixes, as well as for novel GMO constructs. PMID- 12083281 TI - Prevalence of childhood disintegrative disorder. AB - The prevalence of childhood disintegrative disorder (CDD) is unknown. In this study, 32 epidemiological surveys of autism and pervasive developmental disorders published in English language journals since 1966 were reviewed. Four surveys yielded estimates for CDD ranging from 1.1 to 6.4 per 100,000 subjects. A pooled estimate across these four surveys is 1.7 per 100,000 (95 percent Confidence Interval: 0.6-3.8 per 100,000). The conclusion is that CDD is very rare and its prevalence is 60 times less than that for autistic disorder, assuming a prevalence of 10 per 10,000 for autism. If a rate of 30 per 10,000 is taken for all PDDs, only one child out of 175 children with a PDD diagnosis would, on average, meet criteria for CDD. PMID- 12083282 TI - Parent information and direct observation in the diagnosis of pervasive and specific developmental disorders. AB - Children with autism and children with a severe specific receptive language disorder show clear deficits in communicative language skills and social relationships. In this study the usefulness of a standardized parent interview (ADI-R) and a standardized observation schedule (ADOS-G) for the differential diagnosis of these two groups was assessed. Eleven children with early infantile autism and 16 children with a specific receptive language disorder participated. The parent interview was conducted with all parents and the observation schedule was administered to all children. Ten out of 11 children with autism were correctly classified as having autism on the ADI-R and the ADOS-G. One child with a receptive language disorder was falsely classified as having autism on the ADI R, and none on the ADOS-G. Parent interview provides extensive information on the developmental course of the child. Direct observation gives an overview of actual relevant behavioural problems. The two instruments are complementary in the diagnosis of developmental disorders. PMID- 12083283 TI - Developing pretend play in children with autism: a case study. AB - A classroom-based intervention study aimed to explore whether it was possible to teach children with autism and additional learning difficulties to use symbolic pretend play. Five children with autism were involved in a 4 month intervention that used structure, affect and repetition. The intervention progressively faded out the structuring over three phases. All the children were able to use some symbolic acts within play. The study suggests that some of the symbolic play was not the result of replicating previously modelled examples but was spontaneous and novel. PMID- 12083284 TI - A comparison of the effects of structured play and facilitated play approaches on preschoolers with autism. A case study. AB - This study compared the effect of structured play (SP) and facilitated play (FP) in promoting spontaneity and responsiveness in communication and play behaviors in children with autism. SP is characterized by the use of mass practice trials under the instruction of the experimenter while FP incorporated incidental teaching with multiple exemplars. Eight preschoolers participated in a crossover design of both play conditions. Gains in appropriate communication and play were observed across both treatments. Respondent communicative acts occurred more frequently across all participants during SP compared with FP. The preliminary results indicate an interaction between the mental age of the child and the teaching paradigm used. The need to match treatment goals and specific methods of the play intervention to the skill profile of participants is discussed. PMID- 12083285 TI - Body mass index in male and female children with infantile autism. AB - Morphometry, the measurement of forms, is an ancient practice. Recently, evidence has grown to support the notion that aberrant neurodevelopment may play a role in the pathophysiology of autism. Is the body, like the brain, affected by abnormal development in these patients? The aim of this study was to evaluate body mass index (BMI) of children with infantile autism, by comparing the BMI of 117 children with infantile autism with the corresponding BMI percentiles in an age- and sex-matched reference population. The BMI distribution of the male, but not female, children with infantile autism was significantly lower than that of the age-matched reference population. There was no evidence that BMI was associated with intelligence or socioeconomic status among children with infantile autism. PMID- 12083286 TI - Smooth muscle contraction by small GTPase Rho. AB - Abnormal contraction of vascular smooth muscle contributes to a variety of diseases such as hypertension and vasospasm in coronary and cerebral arteries. An increment in a cytoplasmic Ca2+ concentration is the key event in smooth muscle contraction. However, smooth muscle contraction is modified upon the stimulation by agonists as well as in some pathophysiological situations in Ca(2+) independent mechanism. The molecular mechanism underlying this modulation was not elucidated. Recent studies have shown the important role of small GTPase Rho and its effector, Rho-associated kinase (Rho-kinase)/ROK/ROCK in Ca(2+)-independent regulation of smooth muscle contraction. The Rho/Rho-kinase pathway modulates the phosphorylation level of myosin light chain (MLC) of myosin II, mainly through the inhibition of myosin phosphatase, and contributes to the agonist-induced Ca(2+)-sensitization in smooth muscle contraction. The Rho/Rho-kinase pathway is involved in the pathogenesis of hypertension, vasospasm and arteriosclerosis, and is a potent target of new therapies for these diseases. PMID- 12083288 TI - Influence of choking and arm lock technique in judo on the acoustic reflex threshold (art) in healthy well-trained male and female judoka. AB - The objective of this controlled parallel group study was to assess the effects of standardized choke holds (test) and arm lock techniques (controls) in on the acoustic hearing threshold. 104 (test group, 32 female subjects and 72 male subjects, mean age = 28.0 years, SD = 7.9 years) and 51 experienced judoka (controls. 21 female subjects, 30 male subjects; mean age = 26.8 years; SD = 13.2 years) participated. Acoustic reflex thresholds (ART [dB]) were measured separately before and after each manoeuvre both for air and bone conduction of the right and left side. The difference Dart of the ART before and after a manoeuvre (Dart = ARTbefore - ARTafter) was calculated. Data were presented descriptively and nonparametric statistics was applied for nonrelated (Kruskal Wallis ANOVA) or related samples (Friedman ANOVA). Wilcoxon tests were used for pre/post comparisons of original ART values. The effect of choking on Dart was significantly different from the effect of the arm lock technique on Dart independent of the experimental condition. A significant influence of applied frequencies on Dart was ascertained if a choking technique was used. For all frequency ranges applied a highly significant improvement of the ART after choking was found. With regard to bone conduction thresholds increased by an average of 6.1 dB and for air conduction the average increase was 4.9 dB. On the contrary, arm locks induced a slight mean deterioration of the ART for bone conduction of 1.8 dB. The ART for bone conduction also showed a trend towards a reduction after arm locks with a mean decrease of about 1.2 dB. In conclusion, standardized choking manoeuvres reduced the ART corresponding to an improved hearing both with regard to air and bone conduction. Such an effect on hearing ability was not found for arm lock techniques. PMID- 12083287 TI - Effect of lactase preparations in asymptomatic individuals with lactase deficiency--gastric digestion of lactose and breath hydrogen analysis. AB - We compared two lactase preparations derived from Aspergillus orizae (AOL) and Penicillinase multicolor (PML) for stability in the stomach and overall enzymatic activity in 10 asymptomatic subjects with lactase deficiency. The subjects were given 10,000 FCC units of either AOL or PML 30 min prior to or simultaneously with 300 ml of milk. Gastric juice was withdrawn through a nasogastric tube immediately after and every 15 min for 60 min, and breath was sampled before and every 15 min for 6 h after the milk ingestion. When lactase was given simultaneously with the milk, gastric juice lactase activity and galactose concentration were significantly higher than the control levels. When lactase preparations were given 30 min prior to the milk, neither lactase activity nor galactose was detected in the gastric juice. The pH of the gastric juice was about 6.0 after the milk ingestion. Breath hydrogen did not increase when milk was ingested simultaneously with enzymes, but did increase if enzymes were given 30 min prior to milk ingestion. There were no significant differences in lactase activity, galactose concentration in gastric juice, or breath hydrogen when AOL and PML were compared. In conclusion, with exogenous lactase, digestion of lactose begins in the stomach when pH is raised to 6.0 by the buffering action of milk. Lactase preparations are effective assessed by breath hydrogen analysis in asymptomatic individuals with lactase deficiency if the enzymes are given simultaneously with milk. PMID- 12083289 TI - Effect of exercise testing protocol on the relationship between minute ventilation and carbon dioxide production. AB - BACKGROUND: The effect of exercise protocol on the slope of the relationship between minute ventilation and carbon dioxide production (deltaVE/deltaVCO2) has not fully been studied. METHODS: Twenty-five healthy volunteers performed two sessions of incremental bicycle exercise, one with a rapidly increasing staged (RIS: 25W every minute) protocol and another with a slowly increasing staged (SIS: 25W every 3 minutes) protocol to calculate the deltaVE/deltaVCO2. Six of the subjects also participated in sessions of steady state exercise (SSE) test various work rates. RESULTS: The deltaVE/deltaVCO2 was significantly lower when the RIS protocol was used than when the SIS protocol was used (23.3 +/- 2.3 vs. 24.9 +/- 1.8, p < 0.001). Data from the subjects who also underwent SSE sessions revealed that the deltaVE/deltaVCO2 from the SSE protocols was greater than that from the RIS protocol and identical with that from the SIS protocol (125.1 +/- 3.7. 20.6 +/- 1.7, and 24.0 +/- 2.7, respectively, p < 0.05), while arterial partial pressure of carbon dioxide during exercise was not different between the protocols (43.0 +/- 3.3. 39.9 +/- 2.1, and 40.8 +/- 2.6, respectively, n.s.). C ONCLUSIONS: Care must be taken in the interpretation of the values of deltaVE/deltaVCC2, as they are influenced by the difference in exercise protocol. PMID- 12083290 TI - Microglia: immunoregulatory cells in the central nervous system. PMID- 12083291 TI - A perspective on the diagnosis of preterm labor. AB - Over the past few decades there has not been any significant reduction in the rate of preterm birth. As we enter into a new millennium, prematurity is still the leading cause of perinatal morbidity and mortality in the non-anomalous infant. The only major contributions in reducing perinatal mortality in the preterm infant are those in the field of neonatology, while the area of preterm birth prevention has lagged far behind. This article highlights the research and clinical challenges that stand in the way of making a timely yet accurate diagnosis of preterm labor. In addition, possible pathways in the evolution of preterm labor are discussed with emphasis on their significance to diagnosis and treatment. PMID- 12083292 TI - Electronic fetal monitoring of the preterm fetus. AB - Following an analysis of the literature, this article reviews the current practices, guidelines, and recommendations for electronic fetal monitoring of the preterm fetus. The physiologic aspects of fetal heart rate control, electronic fetal monitor parameters, and differences between the preterm and term fetus are discussed. Clearly identified is the need for clinical research regarding electronic fetal monitoring of the preterm fetus to establish evidence-based practice guidelines. PMID- 12083293 TI - Treatment of preterm labor. AB - The ultimate goal of treating preterm labor is to prolong the pregnancy long enough to decrease the incidence of neonatal mortality and morbidity associated with prematurity, while minimizing maternal and fetal risks. There are many controversies in treating preterm labor. Much of this controversy stems from the difficulty in establishing efficacy and safety of interventions and uncertainty of the diagnosis of preterm labor. This article outlines conventional measures and tocolytic therapy directed at prolonging the pregnancy. A review of the effect of tocolytic agents, administration, side effects, and nursing interventions is included. Key words: preterm labor treatment, PMID- 12083294 TI - Implementing preterm labor guidelines: a collaborative care improvement process. AB - Health care organizations today are being challenged to deliver care that is cost effective, satisfying to patients, and based on quality outcomes. Urgency created by inadequate bed capacity as well as financial opportunity prompted United Hospital's Birth Center to launch care improvement activities aimed at assessing appropriateness of antepartal length of stay. Collaboration between all members of the health care team enabled a steering committee to implement evidence-based provider practice guidelines targeting variance around preterm labor management. Other multidisciplinary strategies implemented include a home care prescreening process, case management, and establishment of a peer review process. Within the 1-year care improvement process, the Birth Center successfully decreased the length of stay for preterm labor patients from 6.9 days to 5.3 days. This article describes one institution's efforts to improve care by implementing guidelines for the inpatient management of preterm labor. PMID- 12083295 TI - Musculoskeletal implications of preterm infant positioning in the NICU. AB - Alignment and shaping of the musculoskeletal system occur during each body position that infants experience while in neonatal intensive care. Neonatal nurses and physical therapists can play a major role in designing, modeling, and teaching positioning strategies that promote skeletal integrity, postural control, and sensorimotor organization. Musculoskeletal maturation processes and adverse musculoskeletal consequences are reviewed with an emphasis on clinical implications for neonatal care, discharge teaching, and follow-up. Recommendations are offered for neonatal positioning procedures to prevent extremity malalignment, skull deformities, and gross motor delay. PMID- 12083296 TI - Facilitation of parenting the premature infant within the newborn intensive care unit. AB - A challenge facing neonatal health care providers is to combine technological intensive care for preterm infants with a sensitive and individualized approach facilitating neurobehavioral development while supporting parents in their roles as primary advocates and long-term caregivers. This exploratory study evaluated an individualized nursing intervention based on underlying assumptions of both infant and parent competence. The intervention enhanced parents' ability to appraise their infant's behavior critically and respond in a supportive manner. Study findings provide valuable information on the implications of an individualized nursing intervention both for further research and clinical practice. PMID- 12083297 TI - Model-predictive control of hyperthermia treatments. AB - A model-predictive controller (MPC) of the thermal dose in hyperthermia cancer treatments has been developed and evaluated using simulations with one-point and one-dimensional models of a tumor. The developed controller is the first effort in: 1) the application of feedback control to pulsed, high-temperature hyperthermia treatments; 2) the direct control of the treatment thermal dose rather than the treatment temperatures; and 3) the application of MPC to hyperthermia treatments. Simulations were performed with different blood flow rates in the tumor and constraints on temperatures in normal tissues. The results demonstrate that 1) thermal dose can be controlled in the presence of plant-model mismatch and 2) constraints on the maximum allowable temperatures in normal tissue and/or the pulsed power magnitude can be directly incorporated into MPC and met while delivering the desired thermal dose to the tumor. For relatively high blood flow rates and low transducer surface intensities--factors that limit the range of temperature variations in the tumor, the linear MPC, obtained by piece-wise linearization of the dose-temperature relationship, provides an adequate performance. For large temperature variations, the development of nonlinear MPC is necessary. PMID- 12083298 TI - Optimal resolution of superimposed action potentials. AB - This paper presents a practical algorithm for resolving superimposed action potentials encountered during the decomposition of electromyographic signals. The problem is posed as an optimization problem: to align a set of templates with a given waveform to minimize the euclidean distance between them. The algorithm uses a recursive approach to search all possible discrete-time alignments, starting with the most likely ones and stopping once it can be verified that the optimal alignment has been found. Each candidate solution is aligned to finer than-sampling-interval resolution using interpolation and continuous-time optimization. Both the cases in which the identities of the involved templates are known and not known are considered. Simulations are presented to show that the proposed algorithm is very accurate even for complex superpositions involving three or more similarly shaped templates, destructive interference, and added noise. PMID- 12083299 TI - Wavelet decomposition of cardiovascular signals for baroreceptor function tests in pigs. AB - In this paper, the discrete wavelet transform (DWT) was applied to analyze the fluctuations in RR interval and systolic arterial pressure (SAP) recorded from eight alpha-chloralose anesthetized pigs. Our aim was to characterize the autonomic modulation before and after cardiac autonomic blockade and during baroreflex function tests. The instantaneous power of decomposed low-frequency (LF) and high-frequency (HF) components was used for a time-variant spectral analysis. Our results suggested that transient events and changes in autonomic modulation were detected with high temporal resolution. A nonlinear relationship between RR interval and SAP during pharmacologically induced changes in blood pressure was found, when the superimposed effect of respiratory sinus arrhythmia was removed. In addition, the baroslopes were nearly linear when both the LF and HF components were removed using DWT decomposition. PMID- 12083300 TI - A real-time algorithm for the quantification of blood pressure waveforms. AB - A real-time algorithm for quantification of biological oscillatory signals, such as arterial blood pressure (BP) is proposed which does not require user intervention and works on waveforms complicated by rapid changes in the mean level, frequency, or by the presence of arrhythmia. The algorithm is based on the continous independent assessment of the refractory period (RP). In the first stage, a sample of the signal is band-pass filtered. During the next stage: 1) the local maxima in the filtered signal are identified and their pulse amplitudes (PA) measured on the side opposite to the possible notch position and 2) those maxima whose PA exceeds some threshold are selected and an array of RP values is formed as a fraction of the moving estimate of the interval between successive selected peaks. Finally, the original signal is analyzed by means of two moving averages (MAs) with short and long averaging time intervals. The true peaks are determined as the maxima between intersections of MAs if the peak-to-peak or the intersection-to-intersection intervals since the previous peak and the previous intersection exceed the RP. The algorithm proved to be superior against three commercially available heartbeat detectors yielding an error rate of 0.09%. PMID- 12083301 TI - Wavelet-based ECG compression using dynamic vector quantization with tree codevectors in single codebook. AB - In this paper, we propose a novel vector quantizer (VQ) in the wavelet domain for the compression of electrocardiogram (ECG) signals. A vector called tree vector (TV) is formed first in a novel structure, where wavelet transformed (WT) coefficients in the vector are arranged in the order of a hierarchical tree. Then, the TVs extracted from various WT subbands are collected in one single codebook. This feature is an advantage over traditional WT-VQ methods, where multiple codebooks are needed and are usually designed separately because numerical ranges of coefficient values in various WT subbands are quite different. Finally, a distortion-constrained codebook replenishment mechanism is incorporated into the VQ, where codevectors can be updated dynamically, to guarantee reliable quality of reconstructed ECG waveforms. With the proposed approach both visual quality and the objective quality in terms of the percent of root-mean-square difference (PRD) are excellent even in a very low bit rate. For the entire 48 records of Lead II ECG data in the MIT/BIH database, an average PRD of 7.3% at 146 b/s is obtained. For the same test data under consideration, the proposed method outperforms many recently published ones, including the best one known as the set partitioning in hierarchical trees. PMID- 12083302 TI - Stimulus artifact cancellation in the serosal recordings of gastric myoelectric activity using wavelet transform. AB - Previous studies have shown that electrical stimulation of the stomach (i.e., gastric pacing) with appropriate parameters is a promising method for treatment of gastroparetic patients. The recording of gastric myoelectric activity (GMA) by serosal electrodes is often used to evaluate the effect of stimulation. However, the major problem with the measurement of GMA during gastric pacing is the stimulus artifacts which are often superimposed on the serosal recording and make analysis difficult. The frequency-domain adaptive filter has been used to reduce the stimulus artifacts but only with limited success. This paper describes a wavelet transform-based method for the reduction of stimulus artifacts in the serosal recordings of GMA. The key of this method lies in the use of the fuzzy set theory to select the stimulus artifact-related modulus maxima in the wavelet domain. Both quantitative and qualitative measures show that significant stimulus artifact cancellation was achieved through a series of computer simulations. Results from both single- and multichannel serosally recorded myoelectric signals during gastric pacing are presented to demonstrate the efficiency of the proposed method for the cancellation of stimulus artifacts. PMID- 12083303 TI - A floating-point digital receiver for MRI. AB - A magnetic resonance imaging (MRI) system requires the highest possible signal fidelity and stability for clinical applications. Quadrature analog receivers have problems with channel matching, dc offset and analog-to-digital linearity. Fixed-point digital receivers (DRs) reduce all of these problems. We have demonstrated that a floating-point DR using large (order 124 to 512) FIR low-pass filters also overcomes these problems, automatically provides long word length and has low latency between signals. A preloaded table of finite impuls response (FIR) filter coefficients provides fast switching between one of 129 different one-stage and two-stage multrate FIR low-pass filters with bandwidths between 4 KHz and 125 KHz. This design has been implemented on a dual channel circuit board for a commercial MRI system. PMID- 12083304 TI - A novel design of thermal anomaly for mammary gland tumor phantom for microwave radiometer. AB - Microwave radiometry is the spectral measurement technique of resolving electromagnetic radiation of all matters which temperature is above absolute zero. This technique utilizes the electromagnetic noise field generated by a thermal volume similar to a mechanism existing in biological tissues. One particular application of microwave radiometry is for analyzing temperature differentials of inside of human body to detect and diagnose some crucial pathological conditions. For the general evaluation of a microwave radiometer, we propose a new type of phantom containing a mammary gland tumor imitator by considering biological heat diffusion effects propagated by a real tumor. Theoretical researches of human tumor revealed the fact that temperature distribution of tissues around a tumor formed a Gaussian statistics. To comply with the physiological property of the real tumor, we built a mammary gland tumor imitator composed of two parts (pseudotumor and thermal anomaly) and observed its temperature distribution when it was placed inside a phantom. Our results showed that the thermal properties of tumor imitator well agreed with heat-transfer properties of a real tumor and the proportional linear relationship existed between the location of tumor imitator and the intensity of radiometer measurements. From this relationship, we could also estimate several parameters related with our phantom, such as the minimum detectable size and maximum detectable depth of a tumor imitator. PMID- 12083305 TI - Simulation of human respiration in fMRI with a mechanical model. AB - Obtaining functional magnetic resonance images of the brain is a challenging measurement process having a low characteristic signal-to-noise ratio. Images contain various forms of noise, including those induced by physiologic processes. One of the prevalent disturbances is hypothesized to result from susceptibility fluctuations caused by abdominal volume changes during respiration. To test this hypothesis and characterize the contribution of respiration noise to both magnitude and phase images, a mechanical model of a respiring human was constructed. The model was tested by comparing data from the model with that of a resting human. Power spectrum analyses show that the model induces both phase and magnitude disturbances similar to those in the human. The disturbances are directly related to the frequency of the respiration, with the noise most prevalent in the phase images. Though magnitude image noise is hard to identify in the human, the manikin demonstrates the presence of this disturbance. The construction of the manikin rules out motion as the primary source of the observed fluctuations and variation of the electrical properties of the manikin also indicates that signal fluctuations are not primarily due to eddy currents. Therefore, the changes are most probably induced by bulk susceptibility changes correlating with respiration. PMID- 12083306 TI - Quantification of 3-D field effects during 2-D microwave imaging. AB - Two-dimensional (2-D) approaches to microwave imaging have dominated the research landscape primarily due to the moderate levels of measurement data, data acquisition time, and computational costs required. Three-dimensional (3-D) approaches have been investigated in simulation, phantom, and animal experiments. While 3-D approaches are certainly important in terms of the potential to improve image quality, their associated costs are significant at this time. In addition, benchmarks are needed to evaluate these new generation systems as more 3-D methods begin to appear. In this paper, we present a systematic series of experiments which assess the capability of our 2-D system to image classical 3-D geometries. We demonstrate where current methods suffer from 3-D effects but also identify situations where they remain quite useful. Comparisons between reconstructions utilizing phantom measurements and simulated 3-D data are also shown to validate the results. These findings suggest that for certain biomedical applications, 2-D approaches remain quite attractive. PMID- 12083307 TI - Two-dimensional mapping of impedance magnetocardiograms. AB - A new method for measuring two-dimensional (2-D) impedance magnetocardiograms (I MCGs) and magnetocardiograms (MCGs) above the heart simultaneously, has been developed. The I-MCG's and MCG's signals are recorded by using a superconducting interference device (SQUID) system. To measure the I-MCG and MCG signals, four first-order gradiometers with an 18-mm diameter and a 50-mm baseline were used. The SQUIDs are driven by a flux-locked-loop circuit with a frequency range higher than that of an ac-current (40 kHz) with constant amplitude passed through a subject. The output of the circuit is filtered through two circuits: one for measuring the I-MCG signals and one for measuring the MCG signals. The I-MCG signals are obtained by demodulating the magnetic field, which is detected by the gradiometers, at the frequency of the ac current. As a result, the I-MCG signal reflects the change in spatial distribution of conductivity caused by the movement of the heart muscle and blood volume. A contour map of the 2-D I-MCG signals showed the largest signals occur above the right ventricle and right atrium. In a corresponding current-arrow map, it was found that the large current arrows occurred above the right side of the right ventricle. Furthermore, it was found that the systole and diastole timings obtained from the first-derivative I MCG signal and the phonocardiogram were different. These results show that primitive 2-D I-MCG signal can provide much physiological information on the circulatory movement of the heart. PMID- 12083308 TI - Heart rate indication using musical data. AB - Heart rate data reflects various physiological states such as biological workload, stress at work and concentration on tasks, drowsiness and the active state of the autonomic nervous system. This paper proposes system to indicate heart rate using musical data. Music changes physiological states for the better by relaxing people, or contributing to patient treatment through music therapy. Information in the form of music is advantageous because it does not hinder work as does verbal information and it contains more information than warning noises. We introduce and evaluate a prototype heart rate indication system and describe evaluation results of biofeedback effects on the worker during mental workload. The prototype system sequentially inputs the instantaneous heart rate into the computer, converts the data into musical instrument digital interface, the digital music format, and outputs it from the sound source. PMID- 12083309 TI - Study of features based on nonlinear dynamical modeling in ECG arrhythmia detection and classification. AB - We present a study of the nonlinear dynamics of electrocardiogram (ECG) signals for arrhythmia characterization. The correlation dimension and largest Lyapunov exponent are used to model the chaotic nature of five different classes of ECG signals. The model parameters are evaluated for a large number of real ECG signals within each class and the results are reported. The presented algorithms allow automatic calculation of the features. The statistical analysis of the calculated features indicates that they differ significantly between normal heart rhythm and the different arrhythmia types and, hence, can be rather useful in ECG arrhythmia detection. On the other hand, the results indicate that the discrimination between different arrhythmia types is difficult using such features. The results of this work are supported by statistical analysis that provides a clear outline for the potential uses and limitations of these features. PMID- 12083310 TI - A wavelet-based heart rate variability analysis for the study of nonsustained ventricular tachycardia. AB - It has been reported that the sympathovagal balance (SB) can be quantified by heart rate (HR) via the low-frequency (LF) to high-frequency (HF) spectral power ratio LF/HF. In this paper, an investigation of the relationship between the autonomic nervous system (ANS) and non-sustained ventricular tachycardia (NSVT) is presented. A wavelet transform (WT)-based approach for short-time heart rate variability (HRV) assessments is proposed for this aspect of analysis. The study was conducted on an RR-interval database consisting of 87 NSVT, 61 ischemic and five normal episodes. First, instantaneous SB estimates were generated by the proposed method. Then, waveforms of the WT-based SB evolutions were quantitatively examined. Numerical results showed that while a majority of SB waveforms (about 71%) derived from the non-NSVT population (i.e., ischemic and normal) appeared to come near oscillating with certain fixed levels, approximate 75% of SB evolutions underwent significantly rapid increases prior to the onset of NSVT, suggesting that an abrupt sympathovagal imbalance might partly account for the occurrence of NSVT. PMID- 12083311 TI - Source localization using a current-density minimization approach. AB - Determining the location of cortical activity from electroencephalographic (EEG) data is important clinically. In this paper, a method is presented which uses the powerful optimization method of simulated annealing in conjunction with a finite element-based model of the search domain for single-time slice solution of the EEG-inverse problem. The algorithm highlights a new objective function based on the current-density boundary integral associated with the finite-element formulation as the basis for parameter optimization. In two-dimensional experiments in a shallow tank containing saline, single dipoles are located within 2 mm. Simulations studying the algorithms response to structured noise are also presented. The new objective function is shown to take advantage of the natural framework associated with finite-elements and the results suggest that the approach is capable of resolving dipole locations in simulations and experiments. PMID- 12083312 TI - Numerical computation of impedances of a human tooth for estimation of the root canal length. AB - Alternative current (ac) current conduction through a human tooth has been investigated through numerical simulation. Numerical calculation of impedance between the file (electrode) inserted in the root canal of the tooth and the outer electrode enables investigation of the impedance method used in dentistry for evaluation of the root canal length. Simulations confirm the improved sensitivity of the impedance method using the results of multiple frequency measurements. PMID- 12083313 TI - Serum depletion and bioavailability of lutein in type I diabetic patients. AB - BACKGROUND: Lutein, a non-provitamin A carotenoid, is frequently consumed in the human diet. It is distributed preferentially in certain human tissues (i. e., retina) and shows a high antioxidant activity. Type 1 diabetic patients have been considered to be at risk of increased oxidative stress that may contribute to accelerated atherogenesis and to the microangiopathic complications of the disease. AIM OF THE STUDY: To assess the influence of type 1 diabetes mellitus on the serum depletion rate and bioavailability of lutein. SUBJECTS AND METHODS: Ten type 1 diabetics and eight controls consumed a low carotenoid diet for 21 days and the bioavailability study was performed in 7 diabetics and 5 controls on day 15 with the administration of a capsule of lutein esters from marigold extract. Samples were collected at baseline and on days 1, 2, 3, 6, 11, 15 (eight times during 9 h), 16, 17 and 21. Lutein and other carotenoids were determined by HPLC in triglyceride-rich lipoprotein (TRL) fractions and plasma or serum. RESULTS: Serum depletion curve, area of concentrations under time curve (AUC) and final concentration percentages were similar in diabetics and controls. In the bioavailability study, all-trans-lutein increased in both groups and AUC, maxima concentrations in TRL and serum and time required for maxima concentration in serum were similar in diabetics and controls. CONCLUSIONS: These data suggest that in the group of patients assessed, type 1 diabetes mellitus does not apparently influence the absorption and depletion rate of lutein in serum. PMID- 12083314 TI - Effects of non-esterified stanols in a liquid emulsion on cholesterol absorption and synthesis in hypercholesterolemic men. AB - BACKGROUND Numerous studies have shown that dietary plant sterols (phytosterols and phytostanols) and their esters can decrease cholesterol absorption. However, few researchers have examined the effects of plant sterols on cholesterol absorption and synthesis using stable isotope tracers, instead of relying on endogenous pathway precursors. Further, we have worked with non-esterified lecithin-solubilized stanols as opposed to the more frequently studied esterified sterols and stanols. The vehicle was an oil-in-water liquid emulsion rather than the more common spread vehicle typically employed. AIM OF THE STUDY: To determine the effects of relatively low doses of lecithin-solubilized non-esterified stanols in liquid emulsions on cholesterol absorption and synthesis in mildly hypercholesterolemic subjects. METHODS: In a randomized, double blind crossover design, 12 mildly hypercholesterolemic men received either a free phytostanol supplement (3 g/d in 3 servings) or a control treatment for 3 days. Cholesterol endogenous synthesis rate was determined using the rate of incorporation of deuterium from body water into newly formed cholesterol molecules. Cholesterol absorption at the intestinal level was determined using the dual isotope method using 13C cholesterol injected intravenously and 180 cholesterol given orally. RESULTS: Cholesterol absorption was 55.7 +/- 6.5 % for the control and 33.5 +/- 5.3% for the phytostanol treatment. This massive reduction of the cholesterol absorption did not induce, on average, a difference in cholesterol endogenous synthesis which was measured at 0.074 +/- 0.0015 pool/d for plant sterols and 0.0736 +/- 0.0015 pool/d for controls (p > 0.05). CONCLUSIONS: The results demonstrated that lecithin-solubilized stanols administrated during a short period of time (3 days) in an oil-in-water emulsion can dramatically decrease cholesterol absorption, without a consistent, concomitant increase in synthesis, which is highly suggestive of effective LDL cholesterol lowering. The effects of synthesis should be verified in a longer study with more subjects. PMID- 12083315 TI - Use of BMI as a measure of overweight and obesity in a field study on 5-7 year old children. AB - OBJECTIVE: The present field study examines the use of BMI in comparison with estimates of percent fat mass to screen for overweight and obesity in children. DESIGN: Cross-sectional field study. SETTING: Four waves of children 1996-1999 at Kiel, North West Germany. SUBJECTS: A representative large sample of 2,286 5-7 year old children representing 40% of the total child population examined by school physicians within the same period in Kiel. MAIN OUTCOME MEASURES: BMI was compared with anthropometric measures (TSF, BSF, SIF, SSF) and bioelectrical impedance analysis (BIA). The 90th and 97th BMI percentiles were used as cut offs for overweight and obesity, respectively. RESULTS: BMI reached a low sensitivity to identify overweight children when compared with the two estimates of % FM (0.60 to 0.78 for girls, 0.71 to 0.82 for boys, respectively). The specificity of BMI was 93 to 95 %. By contrast, BMI reached higher sensitivity to screen for obese children of 0.83 to 0.85 for boys and 0.62 to 0.80 for girls at a concomitant specificity of 0.95 to 0.98 for boys and 0.96 to 0.97 for girls as defined by assessment of body fat mass. Comparing nutritional status of overweight children classified as overweight or non overweight by BMI shows that BMI only identified obese but not-overweight children. CONCLUSION: BMI can be used to screen for obese children. In contrast BMI has a poor sensitivity to screen for overweight children. Body composition analysis should be used to screen for children at risk of becoming obese. PMID- 12083316 TI - Hyperhomocysteinemia, and low intakes of folic acid and vitamin B12 in urban North India. AB - BACKGROUND AND AIM: An adverse coronary risk profile has been reported amongst rural-to-urban migrant population living in urban slums undergoing stressful socio-economic transition. These individuals are likely to have low intakes of folic acid and vitamin B12, which may have an adverse impact on serum levels of homocysteine (Hcy). To test this hypothesis, we studied serum levels of Hcy in subjects living in an urban slum of North India and healthy subjects from urban nonslum area. METHODS: Group I consisted of 46 subjects (22 males and 24 females) living in an urban slum, while group II consisted of healthy subjects (n = 26, 13 males and 13 females) living in the adjacent non-slum area. Anthropometric measurements, biochemical profile (fasting blood glucose, total cholesterol, serum triglycerides, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol) and fasting serum levels of Hcy were measured. Dietary intakes of folic acid, vitamin B12, vitamin B1, and iron were calculated by the 24-hour dietary recall method. Serum levels of Hcy were correlated with dietary intakes of nutrients, anthropometry, and metabolic variables. RESULTS: Sex adjusted serum levels of Hcy in micromol/L (Mean +/- SD) were high, though statistically comparable, in both the groups (group I: 20.8 +/- 5.9 and group II: 23.2 +/- 5.9). Overall, higher than normal serum levels of Hcy (> 15 micromol/L) were recorded in 84% of the subjects. A substantial proportion of subjects in both groups had daily nutrient intakes below that recommended for the Asian Indian population (folic acid: 93.4% in group I and 96.7% in group II, vitamin B12: 76.1 % in group I and 88.4% in group II). However, between the two groups, average daily dietary intakes of both the nutrients were statistically comparable. As compared to non-vegetarians, vegetarians showed lower intakes of folic acid (p < 0.01) and vitamin B12 (p < 0.01) in both groups. On multivariate linear regression analysis with serum Hcy as the response variable and vegetarian/non-vegetarian status and sex (male/female) as predictor variables, higher serum levels of Hcy were observed in vegetarians vs non-vegetarians (1 = 4.6, p < 0.05) and males vs females (beta = 5.3, p < 0.01). CONCLUSIONS: Low intakes of folic acid and vitamin B12, and hyperhomocysteinemia, in both the healthy population living in urban slums and adjacent urban non-slum areas, are important observations for the prevention of nutritional and cardiovascular diseases in the Indian subcontinent. PMID- 12083317 TI - Biotinidase catalyzes debiotinylation of histones. AB - BACKGROUND: Posttranslational modifications of histones play important roles in processes such as regulation of gene expression and DNA repair. Recently, evidence has been provided that histones in human cells are modified by covalent attachment of biotin. AIM OF THE STUDY To determine whether the reverse process (debiotinylation of histones) occurs in biological samples and whether debiotinylation is an enzyme-mediated process; and to characterize the enzyme that mediates debiotinylation of histones. METHODS: Plasma and lymphocytes from healthy adults and a biotinidase-deficient patient were used as sources of debiotinylating enzymes. Debiotinylation of histones by plasma and lymphocyte proteins was measured using a colorimetric 96-well plate assay. RESULTS: Histones were debiotinylated rapidly if incubated with human plasma or lysates of lymphocytes. The following observations are consistent with the hypothesis that debiotinylation is an enzyme-mediated process: (i) Hydrolysis was slower at 4 degrees C compared to 37 degrees C; (ii) debiotinylating activity was destroyed when biological samples were heated at 90 degrees C for 30 min preceding incubation with biotinylated histones; and (iii) rates of debiotinylation were pH dependent. Rates of histone debiotinylation were significantly decreased in biotinidase-deficient samples. CONCLUSION: Debiotinylation of histones in human samples is an enzyme-mediated process that is at least partly catalyzed by biotinidase. PMID- 12083318 TI - Detection of ligands for selectins in the oligosaccharide fraction of human milk. AB - BACKGROUND: Human milk contains a large variety of oligosaccharides which show struc tural similarities with ligands for selectins, a family of cell adhesion molecules which are involved in many cell-cell interactions. AIM OF THE STUDY: Due to their structural similarity with selectin ligands, human milk oligosaccharides were labelled with phosphatidyl ethanolamine to be able to investigate specific effects of antibodies against carbohydrate epitopes. METHODS: Various monoclonal antibodies against physiological selectin ligands were used to determine whether epitopes within human milk oligosaccharides are recognized. Oligosaccharides were isolated from human milk, transferred into neoglycolipids and separated using high performance thin layer chromatography prior to incubation with monoclonal antibodies for the selectin ligands sialyl Lewis a (sLe a), sialyl-Lewis x (sLe x), Lewis x (Le x) and Lewis y (Le y) after Western blotting. Fast atom bombardment-mass spectrometry was then used to identify antibody-binding compounds. Results In the immunoassays all ligand epitopes except for Le y were detected in the oligosaccharide fraction of human milk. Anti-sLe a showed the most distinct reaction with N-acetylneuraminic acid containing neoglycolipids of which two were identified as neuraminylfucosyl-lacto N-hexaose and neuraminyl-lacto-N-tetraose. Such oligosaccharides as well as similar structures are present in relatively high concentrations in human milk. CONCLUSIONS: The presence of sialyl-Lewis ligands on milk oligosaccharides together with their abundancy in human milk may suggest that they could be selectin ligands and they may be part of inflammatory processes. PMID- 12083319 TI - Isothiocyanates as cancer chemopreventive agents: their biological activities and metabolism in rodents and humans. AB - Isothiocyanates (ITCs) are a group of naturally occurring compounds that occur as thioglucoside conjugates, termed glucosinolates, in plants and cruciferous vegetables such as watercress, Brussels sprouts, broccoli, cabbage, kai choi, kale, horseradish, radish and turnip. ITCs inhibit the development of tumors in many of the experimental models investigated, and are being investigated as possible chemopreventive agents for specific human cancers. The goal of this review is to provide a mechanistic understanding for the biological activities of ITCs and to relate the metabolism of ITCs to their action as chemopreventive agents. In vivo animal studies have been conducted to address issues of tissue disposition, pharmacokinetics, and metabolism of ITCs. Methods for analysis of ITCs and their metabolites in urine and plasma have been developed. The metabolism of several naturally occurring ITCs as constituents of foodstuffs or as drugs has also been investigated in human studies. Finally, based on recent epidemiological studies, the role of dietary consumption of vegetables containing ITCs in prevention of human cancers and human cancer susceptibility is discussed. PMID- 12083320 TI - Evaluation of drug interactions with P-glycoprotein in drug discovery: in vitro assessment of the potential for drug-drug interactions with P-glycoprotein. AB - The pharmacological effects of a drug are highly dependent on the absorption, metabolism, elimination, and distribution of the drug. In the past few years it has become apparent that transport proteins play a major role in regulating the distribution, elimination and metabolism of some drugs. As a consequence of our new understanding of the influence of transport proteins on the pharmacokinetic and pharmacodynamic behavior of drugs, increasing attention has been focused on the potential for drug-drug interactions arising from interactions with drug transport proteins. The efflux transporter P-glycoprotein (P-gp) has received the most attention with regard to its role in restricting drug absorption and distribution and as a potential source for variability in drug pharmacokinetics and pharmacodynamics. This review will focus on the evaluation of drug candidates to assess the potential for drug interactions at the level of P-gp. We will discuss the role of P-gp in drug disposition, the biochemistry of P-gp efflux as it relates to model systems to study drug interactions with P-gp, and the implementation of P-gp assay models within the drug discovery process. PMID- 12083322 TI - Cytochrome P450 pharmacogenetics in drug development: in vitro studies and clinical consequences. AB - Members of the human cytochrome P450 (CYP) superfamily play a role in the metabolism of many drugs and several of them, CYP2D6, CYP2C9 and CYP2C19, have been shown to be polymorphic as a result of single nucleotide polymorphisms (SNPs), gene deletions, and gene duplications. These polymorphisms can impact the pharmacokinetics (PK), metabolism, safety and efficacy of drugs, and because of the availability of automation, genotyped human tissue, recombinant CYP preparations (rCYPs) and reagents, most pharmaceutical companies have increasingly screened out compounds that are metabolized solely by polymorphic CYPs. In the absence of suitable animal models, it has been widely accepted that such in vitro data are useful because one can obtain information prior to dosing in man and select the most appropriate clinical studies with prospectively genotyped and phenotyped subjects. Overall, current trends in the industry have been fueled by increased managed healthcare, the desire to minimize the need for therapeutic drug monitoring and CYP genotyping in medical practice, and a very competitive market place. In the past, such paradigms have not been as influential and there are numerous examples of marketed drugs that are metabolized by polymorphic CYPs. PMID- 12083321 TI - Review: metabolism of immunosuppressant drugs. AB - Pharmacokinetic concepts provide a basis for individualization of drug therapy to optimize outcomes of the critical-dose drugs cyclosporine (CsA), tacrolimus (TRL), sirolimus (SRL), and mycophenolate mofetil (MMF). The therapeutic range of a drug-defined as the concentrations at which the desired pharmacologic effect is produced without adverse effects in most patients-is difficult to achieve given the significant inter-and intrapatient variability of the effects of a given concentration of therapeutic agents. Because of the highly variable rates of absorption of immunosuppressive agents and clinical responses to given concentrations in transplant recipients, individualization of drug regimens by using therapeutic drug monitoring (TDM) is essential to optimize pharmacotherapy Assessing proclivity for acute rejection episodes in transplant recipients currently is attempted by estimating drug exposure using the area under the time concentration curve (AUC) for MMF and the average concentration (Cav, the quotient of the AUC and the dosing interval) for CsA. These studies have revealed that low oral bioavailability was a more important predictor of rejection than was a rapid clearance rate. In addition, the degree of intra-individual variability of AUC values correlated with the development of chronic rejection in renal transplant recipients. Similarly, TDM of MMF requires AUC determinations. Low mycophenolic acid (MPA) exposure, as estimated by the AUC, demonstrates a significant association with an increased risk of an acute renal transplant rejection episode. The AUC0-2 estimate of MPA shows good agreement with the 12-hr AUC estimate from samples obtained during the entire dosing interval. In contrast, trough levels are utilized during treatment with TRL or SRL, potent new immunosuppressive agents that display a pleiotropic array of side effects. Standard body measures, including weight and body mass index, poorly predict the concentration of SRL in whole blood. Large inter- and intra-individual differences displayed in patients also could not be predicted by demographic features or by laboratory parameters. When SRL is given with other immunosuppressive agents such as CsA, which shares with SRL mutual microsomal metabolism by the cytochrome P450 3A system, pharmacokinetic interactions occur, especially when the agents are administered concomitantly. Because of the critical-dose nature of most of the recent generation of immunosuppressive agents, therapeutic drug monitoring is becoming increasingly important in the selection of doses and treatment regimens. PMID- 12083323 TI - Mutagenesis by environmental pollutants and bio-monitoring of environmental mutagens. AB - There is serious concern about the adverse effects of environmental pollutants on human health. Various mutagens, which pollute air, water, and food, possibly induce mutations in humans, and are suspected of causing cancer. Environmental mutagens, such as polycyclic aromatic hydrocarbons (PAH) and heterocyclic amines are known to bind to nucleotides, resulting in the formation of DNA adducts. Some DNA adducts are fixed as mutations through replication of DNA. Reactive oxygen species generated by pollutants also induce the formation of DNA adducts. DNA adducts have been detected as a marker for the exposure of humans and wild life to mutagens. Because of its high sensitivity the 32P-postlabel-thin layer chromatography (TLC) method is widely used for the analysis of DNA adducts formed by PAH and related bulky compounds. However, new systems are required for detecting mutations induced in genomic DNA in vivo to monitor environmental mutagens. Recently, transgenic animals, in which a target gene for detecting mutations is integrated, have been developed. With these transgenic animals, not only mutant frequency but mutation spectra can be determined. We review here recent advances in the detection of DNA adducts formed by environmental pollutants and their application for biological monitoring of environmental mutagens. We also discuss transgenic animals as important tools for evaluating the total mutagenic potential of environmental chemicals. PMID- 12083324 TI - Catechol-O-Methyltransferase (COMT)-mediated methylation metabolism of endogenous bioactive catechols and modulation by endobiotics and xenobiotics: importance in pathophysiology and pathogenesis. AB - The metabolic O-methylation of endogenous catecholamines and other catechols catalyzed by catechol-O-methyltransferase (COMT; EC 2.1.1.6) was first described by Dr. Julix Axelrod and his colleagues almost half a century ago. In the past several years, research interest in this catechol-metabolizing system has been renewed because of its potential pathophysiological and pathogenic significance in estrogen-induced hormonal cancers, in the development of degenerative brain disorders, as well as in the development of cardiovascular diseases. In this review paper, I provide a brief overview of the COMT metabolic system, with particular attentions being paid to the following three areas: (i) the regulation of this catechol-metabolizing system by endogenous regulatory factors (mainly S adenosyl-L-homocysteine and homocysteine) as well as by exogenous factors such as dietary phytochemicals; (ii) decreased metabolic O-methylation of endogenous catecholamines as an important risk factor for the development of neurodegenerative disorders such as Parkinson's and Alzheimer's diseases in the elderly and also as a risk factor for the development of a variety of cardiovascular diseases; and (iii) the relative importance of the COMT-catalyzed O-methylation metabolism of endogenous catechol estrogens in the causation and prevention of estrogen-induced hormonal cancers. Some unifying hypotheses are also discussed in this paper with the hope that they may provide useful mechanistic insights into our understanding of the biological functions that are associated with this important metabolic system. PMID- 12083325 TI - Early shutoff of host protein synthesis in cells infected with herpes simplex viruses. AB - Herpes simplex viruses 1 (HSV-1) and 2 (HSV-2) are capable of suppressing the host cell protein synthesis even without viral gene expression. This phenomenon is known as the early shutoff or as the virion-associated host shutoff (vhs) to emphasize that it is mediated by a component of infecting virions which is a product of the UL41 (vhs) gene. The UL41 encoded protein is a functional tegument protein also present in light (L) particles and is not essential for virus replication. The major product of UL41 gene is a 58 K phosphoprotein. At least two forms of UL41 protein differing in the extent of phosphorylation are present in HSV-1-infected cells. HSV-2 compared to HSV-1 strains display a stronger vhs phenotype. However, in superinfection experiments the less strong vhs phenotype is dominant. UL41 protein triggers disruption of polysomes and rapid degradation of all host and viral mRNAs and blocks a reporter gene expression without other HSVs proteins. The available evidence suggests that UL41 protein is either itself a ribonuclease (RNase) or a subunit of RNase that contains also one or more cellular subunits. UL41 protein is capable of interacting with a transactivator of an alpha-gene, the alpha-transinducing factor (alpha-TIF). Interaction of UL41 protein with alpha-TIF down regulates the UL41 (vhs) gene activity during lytic infection. The possible role of other viral proteins in the shutoff is discussed. PMID- 12083327 TI - Significance of anti-interferon-alpha2 and sICAM-1 activities in the sera of viral hepatitis B and C patients treated with human recombinant interferon alpha2. AB - In this study the presence of an IFN-binding activity in the sera of patients with chronic viral hepatitis B or C treated with rIFN-alpha2 was screened by a radioimmune assay (RIA) using radiolabeled rIFN-alpha2. Incidence of an anti-IFN activitywas compared with hepatitis B virus (HBV) or hepatitis C virus (HCV) serum markers as hepatitis B s antigen (HBsAg), hepatitis B e antigen (HBeAg), antibodies to HBsAg (anti-HBsAg), antibodies to HBeAg (anti-HBeAg), seroconversion, HBV DNA, HCV RNA, and serum soluble intracellular adhesion molecule I (sICAM). Injections (intramuscular) of rIFN-alpha2 caused an anti-rIFN activity formation in 8 (27.6%) of 29 patients with chronic active hepatitis B (CAH-B) and in 8 (30.8%) of 26 patients with chronic active hepatitis C (CAH-C). The presence of the anti-rIFN activity in CAH-B patients correlated frequently with the persistence of HBsAg, HBeAg and HBV-DNA, while its absence was often accompanied by the anti-HBeAg and anti-HBsAg seroconversion, respectively, and HBV-DNA negativity. In two CAH-C patients who became HCV RNA-negative no anti-IFN activity was found. Levels of serum sICAM-1 in CAH-B patients responding to the IFN treatment were higher than those in non-responders or in which the anti-IFN activity was present. The anti-IFN activity may negatively influence the effect of the IFN therapy of CAH-B or CAH-C patients at early stages of the therapy. The appearance of the anti-IFN activity at the end of a long-term IFN therapy does not seem to influence the outcome of the therapy. sICAM-1 may be involved in the process of CAH-B reactivation and IFN-triggered cytotoxicity during the IFN therapy. PMID- 12083326 TI - Human immunodeficiency virus 1 strains resistant to nucleoside inhibitors of reverse transcriptase in isolates from the Czech Republic as monitored by line probe assay and nucleotide sequencing. AB - The genetic resistance to nucleoside inhibitors of the reverse transcriptase (RT) of human immunodeficiency virus I (HIV-1) isolates in the Czech Republic was examined by a line probe assay (LiPA) and nucleotide sequencing. The results of LiPA analysis of 294 blood specimens obtained from 156 patients revealed a high incidence of mutations in the RT gene related to resistance to various drugs (67.3%) in various combinations. Mutations in RT gene (M41L, K70R and T215Y/F) conferring the resistance to zidovudine (ZDV) were most frequent (62.6%), that (M184V) responsible for the resistance to lamivudine (3TC) was less frequent (33.7%), while those linked to the resistance to dideoxyinosine (ddl) and dideoxyinosine together with dideoxycytidine (ddl/ddC) were rather rare (6.5% and 5.1%, respectively). LiPA gave a high rate of uninterpretable results due to codon hybridization failure, especially in HIV-1 isolates of non-B subtype. Thirty-two specimens were analyzed also by direct sequencing of a part of RT gene. The results obtained by LiPA and the sequencing were highly concordant for codons successfully analyzed by both methods, but the sequencing provided information also about the codons that could not be analyzed by LiPA. A high prevalence of resistant strains in the Czech Republic and their heterogeneity justifies a regular HIV-1 resistance testing. LiPA turned out as a fast, powerful and most reliable tool for such a purpose. However, due to an increasing diversity of HIV-1 strains circulating in the Czech Republic, LiPA cannot replace the nucleotide sequence analysis. PMID- 12083328 TI - Efficacy of alpha-interferon therapy of chronic hepatitis patients infected with wild type hepatitis B virus and HBEAG-minus mutant. AB - The aim of this study was to evaluate the efficacy of alpha-interferon (alpha IFN) treatment of 56 chronic hepatitis B (HB) patients positive for HB e antigen (HBeAg), which were previously not treated with alpha-IFN (group A). Seven of them, which did not respond to initial alpha-IFN treatment, were subjected to additional treatment with alpha-IFN (group B). Another 7 patients with chronic HB caused apparently by an HBeAg-minus HB virus (HBV) mutant represented group C. In the alpha-IFN treatment, 5 megaunits (MU) of alpha-IFN were administered subcutaneously three times a week for six months. A trend of improvement of important markers of the disease in the treated patients could be seen with increasing time after completion of the treatment even though it was not statistically significant. In group A, the absence of serum HBV DNA was found in 43% of the patients at the end of the treatment, in 41% 6 months later, and in 46% 12 months later. At the same time intervals group A showed negative HBeAg in 36%, 39% and 46%, positive anti-HBeAg in 36%, 38%, and 46%, negative HBsAg in 9%, 11%, and 14%, and normal level of alanine transaminase (ALT) in 23%, 39%, and 44%, respectively. A trend toward better results of alpha-interferon therapy for the group A patients displaying lower baseline viremia and higher ALT activity could be seen; however, this relationship was not statistically significant. Groups B and C were too small for statistical analysis. Nevertheless, 4 of 7 patients of group B were negative for HBV DNA 12 months after the treatment and HBV DNA was eliminated during the treatment in all patients of group C; however, 3 patients relapsed after the treatment. PMID- 12083329 TI - Evaluation of a dipstick ELISA and a rapid immunochromatographic test for diagnosis of Dengue virus infection. AB - Here we report standardization of a dipstick enzyme-linked immunosorbent assay (ELISA, Dipstick ELISA) and its comparative evaluation with a commercial Rapid PanBio Immunochromatographic test (IC test) for detection of Dengue (DEN) virus specific IgM and IgG antibodies in patient sera. Among crude and purified viral antigens prepared from mouse brains or cell cultures, a DEN virus type 2 antigen purified from cell cultures by sucrose density gradient centrifugation was found superior in terms of the signal/ noise (S/N) ratio in the assay system. The sensitivity of detection of the virus by specific IgM antibody was improved by removal of IgG from patient sera prior to testing. The evaluation of the Dipstick ELISA by use of 156 serum samples revealed an overall accordance of 96% and 93% with the IC test in detection of IgM antibodies to DEN viruses (IgM antibodies) and IgG antibodies to DEN viruses (IgG antibodies), respectively. The sensitivity of the Dipstick ELISA and the IC test with reference to the mu-capture ELISA was 83% and 87%, respectively, with a specificity of 98% in both cases. The sensitivity of the Dipstick ELISA with reference to the IC test in detecting IgM and IgG antibodies was 84% and 94%, respectively, and the specificity of the Dipstick ELISA was 98% and 92%, respectively. PMID- 12083330 TI - Isolation of chikungunya virus from Aedes aegypti mosquitoes collected in the town of Yawat, Pune District, Maharashtra State, India. AB - Chikungunya (CHIK) virus is prevalent throughout Southeast Asia and Africa. It has caused numerous large outbreaks in India. No active or passive surveillance has been carried out since the last epidemic occurring in 1971. During a recent outbreak of Dengue (DEN)-like illness in eastern India, Aedes aegypti mosquitoes collected from the affected area were positive for CHIK virus. Evidence of dual infection with CHIK and DEN typel virus was also obtained. A widely circulating low-virulent CHIK virus is a possible explanation for the epidemiological pattern of the CHIK virus disease in this region. PMID- 12083331 TI - The UL9 ori-binding protein of herpes simplex virus 1: its expression and localization in vero cells. AB - The ori-binding protein (OBP), an early protein which is encoded by the herpes simples virus 1 (HSV-1) UL9 gene and initiates the replication of viral DNA, was expressed in Escherichia coli, purified on an avidin resin and used for preparation of a mouse antiserum to OBP (OBP antiserum). Expression and localization of OBP in HSV-1-infected Vero cells was assessed by reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunofluorescence test. RT-PCR revealed the presence of abundant UL9 transcripts from 3 to 12 hrs post infection (p.i). Traces of UL9 mRNA were detected already at 1.5 hr p.i. The OBP antiserum detected clumps of irregularly shaped structures in the nuclei of infected Vero cells first at 4 hrs p.i. These nuclear structures peaked at 5-6 hrs p.i. and later on (at 8-12 hrs p.i.) they changed into fine granules filling the whole nucleus. PMID- 12083332 TI - Cloning and sequencing of the coat protein gene of tobacco ringspot virus isolates from UK and Iran. AB - The coat protein (CP) gene and the 3' untranslated region (UTR) of genomic RNA 2 of Tobacco ringspot virus (TRSV, the genus Nepovirus, subgroup a) isolates from the UK and Iran were cloned from total viral RNA and sequenced. Comparison of these isolates with an isolate from the USA revealed a high degree of nucleotide and amino acid identity which extends the knowledge of molecular relationships between these three TRSV isolates. The UK isolate shared the highest nucleotide identity (95%) with the US isolate as compared to a lower identity with the Iranian isolate (92%). The highest identity (98%) was found between the UK and US isolates at amino acid level. Comparative analysis of the Iranian, UK and US isolates revealed some differences concerning some members of other subgroups of nepoviruses. For example, the N-terminal FDAYXR and the C-terminal FYGRXS motifs conserved among some nepoviruses, which occur adjacent to each other in folded CP molecules, were not detected in the Iranian, UK or US TSRV isolates. These isolates shared similarity only with Tomato ringspot virus (TomRSV) belonging to the subgroup c of nepoviruses. Another similarity of these isolates with TomRSV and Raspberry ringspot virus (RRSV) was the presence of a 34 nucleotide (nt) long sequence within the 3'-UTR. PMID- 12083333 TI - Characterization of Sri Lanka rabies virus isolates using nucleotide sequence analysis of nucleoprotein gene. AB - Thirty-four suspected rabid brain samples from 2 humans, 24 dogs, 4 cats, 2 mongooses, I jackal and I water buffalo were collected in 1995-1996 in Sri Lanka. Total RNA was extracted directly from brain suspensions and examined using a one step reverse transcription-polymerase chain reaction (RT-PCR) for the rabies virus nucleoprotein (N) gene. Twenty-eight samples were found positive for the virus N gene by RT-PCR and also for the virus antigens by fluorescent antibody (FA) test. Rabies virus isolates obtained from different animal species in different regions of Sri Lanka were genetically homogenous. Sequences of 203 nucleotides (nt)-long RT-PCR products obtained from 16 of 27 samples were found identical. Sequences of 1350 nt of N genes of 14 RT-PCR products were determined. The Sri Lanka isolates under study formed a specific cluster that included also an earlier isolate from India but did not include the known isolates from China, Thailand, Malaysia, Israel, Iran, Oman, Saudi Arabia, Russia, Nepal, Philippines, Japan and from several other countries. These results suggest that one type of rabies virus is circulating among human, dog, cat, mongoose, jackal and water buffalo living near Colombo City and in other five remote regions in Sri Lanka. PMID- 12083334 TI - Identification of rice black streaked dwarf virus in different cereal crops with dwarfing symptoms in China. AB - This report describes isolation of virus particles from plants of rice, maize, wheat and sorghum with symptoms of dwarfing collected from two provinces of China, purification of double-stranded RNA (dsRNA) from the virus particles, and synthesis of full-length cDNAs of genome segments 9 (S9) and 10 (S10) by reverse transcription-polymerase chain reaction (RT-PCR). Sequence analysis showed that the S9 sequences of the Chinese isolates and a Japanese rice black-streaked disease virus (RBSDV) isolate were very similar (89.1-89.6% homology at nucleotide level and 92.3-92.9% and 95.8-98.6% homology at amino acid level for ORF1 and ORF2, respectively). Analogical similarity was found also for the S10 sequences of the isolates under comparison: 93.0-95.4% homology at nucleotide level and 96.2-97.0% homology at amino acid level. However, there was a relatively lower similarity for S9 and S10 segments ofthe Chinese isolates and an Italian maize rough dwarf virus (MRDV) isolate. The phylogenetic analysis indicated that the Chinese isolates that infect rice, maize, wheat and sorghum and cause similar symptoms could represent the same virus species, RBSDV. PMID- 12083335 TI - Proton MR spectroscopy of neurometabolites in hepatic encephalopathy during L ornithine-L-aspartate treatment--results of a pilot study. AB - Hepatic encephalopathy (HE) is associated with typical changes in neurometabolites most likely being caused by an elevated systemic ammonia level. Blood ammonia is a valid over-all biomarker of HE and thus is regularly determined in clinical trials. The neurometabolites affected in HE can be assessed in vivo by proton magnetic spectroscopy. The aim of this study was to show the effect of the ammonia lowering drug L-ornithine-L-aspartate (OA) on the cerebral glutamate+glutamine/creatine (Glu+GLN/Cr) ratio. In an open clinical trial (pilot study), 15 patients with stable HE were treated with an infusion of 40 g OA over 8 h (5 g/h). Immediately before and 6-8 (mean 6.8) h after start of the infusion, spectroscopy of the parietal white matter was performed and arterial blood ammonia quantified. Glu+GLN/Cr-ratios correlated significantly with ammonia data (Spearman's correlation coefficient rs = 0.72, p < 0.001). Likewise, the OA induced changes (versus baseline before infusion) in blood ammonia level and in Glu+GLN/Cr-ratios correlated significantly rs = 0.54, p = 0.0375). Magnetic resonance (MR) spectroscopy assesses the neurometabolite variation present in HE. OA induced changes in cerebral Glu+GLN/ Cr-ratio were significantly correlated with the drug effects on arterial blood ammonia. These pilot data indicate that MR spectroscopy detects a specific biomarker of HE that may reflect the extent of the cerebral alterations associated with the disease. PMID- 12083336 TI - Ischemic cell death: dynamics of delayed secondary energy failure during reperfusion following focal ischemia. AB - Reperfusion injury is believed to contribute to the pathophysiology of ischemic cell death, but the precipitating factors have yet to be completely elucidated. The goal of this study was to examine if reflow-induced secondary energy failure is a component in the events that lead to cell death following increasing periods of middle cerebral artery (MCA) occlusion in Wistar rats. Discrete sections within the MCA distribution were dissected and analyzed for high-energy phosphates and glucose. Regional cerebral blood flow was determined by [14C] iodoantipyrine technique in representative groups. The levels of ATP + P-creatine were initially depressed at the end of the focal ischemia and the concentrations in the penumbra were unchanged for up to 8 h after 2 h of ischemia which contrasts with response in the ischemic core, striatum, and penumbra where the HEP generally recovered to values near those of control only to decrease with increasing periods of reflow. The possibility of a rebound ischemia in secondary energy failure (SEF) was precluded by regional CBF values and concentrations of glucose that were significantly higher than the threshold for an ischemic effect. The depletion of cellular energy stores following SEF strongly indicates that the evolution of infarct during reflow results from loss of ATP and its synthesis. PMID- 12083337 TI - Testosterone treatment attenuates the effects of facial nerve transection on glial fibrillary acidic protein (GFAP) levels in the hamster facial motor nucleus. AB - Testosterone propionate (TP) administration coincident with facial nerve injury accelerates the recovery rate from facial muscle paralysis in the hamster. One mechanism by which TP could augment peripheral nerve regeneration is through glial fibrillary acidic protein (GFAP) regulation in the facial motor nucleus. In a previous study, axotomy alone induces increases in GFAP mRNA. with TP significantly attenuating the axotomy-induced increases in GFAP mRNA. In the present study, immunoblotting techniques were used to extend our previous GFAP mRNA studies to the protein level. Castrated male hamsters were subjected to a right facial nerve transection, with half of the animals receiving subcutaneous implants of 100% crystalline TP. The left facial motor nucleus of each animal served as an internal control. Postoperative survival times include Days 4, 7, and 14. In non-TP-treated animals, facial nerve transections alone increased GFAP levels at all time points, relative to internal controls. As previously observed at the mRNA level, TP treatment attenuated but did not eliminate the axotomy induced increase in GFAP levels at all time points tested. These results suggest that the regulatory actions of gonadal steroids on GFAP expression manifested in parallel at the mRNA/protein levels. PMID- 12083338 TI - Effect of the branched-chain alpha-ketoacids accumulating in maple syrup urine disease on the high molecular weight neurofilament subunit (NF-H) in rat cerebral cortex. AB - In this study we investigated the effects of the branched chain alpha-ketoacids accumulating in maple syrup urine disease (MSUD) on the concentrations of the high molecular weight neurofilament subunit (NF-H) associated with the cytoskeletal fraction of the cerebral cortex of 12-day-old rats. Cortical slices were incubated with alpha-ketoisocaproic acid (KIC), alpha-keto beta methylvaleric acid (KMV) and alpha-ketoisovaleric acid (KIV) at concentrations ranging from 0.5 to 1.0 mM. The cytoskeletal fraction was extracted and the immunoreactivity for phosphorylated and total NF-H was analyzed by immunoblotting. The in vitro 32P incorporation into NF-H was also determined. Results showed that treatment of tissue slices induced with KMV increased Triton insoluble phosphorylated NF-H immunoreactivity, with no alteration in total NF-H immunoreactivity. Furthermore, KIC treatment drastically increased the total amount of NF-H, whereas KIV did not change either phosphorylated or total NF-H immunoreactivity. KMV also increased the in vitro 32P incorporation into NF-H, confirming the highly phosphorylated NF-H levels detected in the immunoblot. These findings demonstrate that KIC and KMV alter the dynamic regulation of NF-H assembly in the cytoskeletal fraction. Therefore we may suggest that cytoskeletal disorganization may be one of the factors associated with the neurodegeneration characteristic of MSUD disease. PMID- 12083339 TI - Selective loss of expression of glutamate GluR2/R3 receptor subunits in cerebellar tissue from a patient with olivopontocerebellar atrophy. AB - Expression of the mRNAs encoding the astrocytic (EAAT1, EAAT2) and neuronal (EAAT3, EAAT4) excitatory amino acid transporters and the AMPA-type glutamate receptor subunits GluR2 and GluR3 was investigated in postmortem cerebellar extracts from a patient with olivopontocerebellar atrophy (OPCA) and in material from three age-matched controls. Decreased expression in the steady state level of EAAT4 mRNA in the OPCA sample was correlated with the selective loss of Purkinje cells. Neuropathological evaluation revealed reactive gliosis and concomitantly increased expression of the mRNA encoding astrocytic glial fibrillary acidic protein (GFAP). Expression of the mRNAs encoding the AMPA receptor subunits GluR2 and GluR3 subunits was found to be decreased in OPCA suggesting that excitotoxic mechanism could play a role in the pathogenesis of the selective neuronal cell death in this disorder. PMID- 12083340 TI - Inhibition of Na+, K+-ATPase activity by the metabolites accumulating in homocystinuria. AB - Homocystinuria is an inborn error of sulfur amino acid metabolism characterized predominantly by vascular and nervous system dysfunction. In this study we determined the in vitro effects of homocysteine and methionine, metabolites which accumulate in homocystinuria, on Na+, K+-ATPase, and Mg2+-ATPase activities in synaptic membranes from the hippocampus of rats. The results showed that both metabolites significantly inhibit Na+, K+-ATPase but not Mg2+-ATPase activity at concentrations usually observed in plasma of homocystinuric patients. Furthermore, incubation of hippocampal homogenates with homocysteine also elicited an inhibition of the enzyme activity which was however prevented by the simultaneous addition of cysteine to the medium. In addition, cysteine or methionine per se did not modify the two enzymatic activities. These findings indicate that oxidation of critical groups in the enzyme may possibly be involved in homocysteine inhibitory effect. Moreover, kinetic studies performed to investigate the interaction between homocysteine and methionine on Na+, K+-ATPase inhibition suggested a common site for the two amino acids in the enzyme. Considering the critical role exerted by Na+, K+-ATPase in brain, it is proposed that the inhibition provoked by homocysteine and methionine on the enzyme activity may be possibly related to the brain dysfunction characteristic of homocystinuria. PMID- 12083342 TI - The treatment of Alzheimer's disease: where have we been, and where are we going? PMID- 12083341 TI - Ganglioside alterations in the central nervous system of rats chronically injected with methylmalonic and propionic acids. AB - Neurological dysfunction and structural cerebral abnormalities are commonly found in patients with methylmalonic and propionic acidemia. However, the mechanisms underlying the neuropathology of these disorders are poorly understood. We have previously demonstrated that methylmalonic and propionic acids induce a significant reduction of ganglioside N-acetylneuraminic acid in the brain of rats subjected to chronic administration of these metabolites. In the present study, we investigated the in vivo effects of chronic administration of methylmalonic (MMA) and propionic (PA) acids (from the 6th to the 28th day of life) on the distribution and composition of gangliosides in the cerebellum and cerebral cortex of rats. Control rats were treated with the same volumes of saline. It was first verified that MMA and PA treatment did not modify body, cerebellum, or cortical weight, nor the ganglioside concentration in the cerebral cortex of the animals. In contrast, a significant reduction in total ganglioside content in the cerebellum of approximately 20-30% and 50% of control levels occurred in rats injected with MMA and PA, respectively. Moreover, chronic MMA and PA administration did not interfere with the ganglioside pattern in the cerebral cortex, whereas the distribution of individual gangliosides was altered in the cerebellum of MMA- and PA-treated animals. Rats injected with MMA demonstrated a marked decrease in GM1 and GD3, whereas chronic PA treatment provoked a significant reduction of all ganglioside species, with the exception of an increase in GM2. Since gangliosides are closely related to the dendritic surface and other neural membranes, indirectly reflecting synaptogenesis, these ganglioside abnormalities may be associated with the brain damage found in methylmalonic and propionic acidemias. PMID- 12083343 TI - Greek validation of the seven-minute screening battery for Alzheimer's disease in the elderly. AB - The increasing prevalence ofAlzheimer's disease (AD) suggests that there is an increasing need for accurate and easily administered screening instruments. The Seven-Minute Screen is a neurocognitive screening battery consisting of four brief tests (enhanced cued recall, temporal orientation, verbal fluency, and clock drawing). We studied 55 outpatients with probable AD, 40 healthy volunteers of comparable age, sex, and education and 31 elderly patients with other neuropsychological disorders. The aim of our study was to determine the validity and reliability of this test. Differences on individual tests were evaluated using the Student t test. (Recall: 6.4 +/- 5.02/15.38 +/- 0.95; Orientation: 48.76 +/- 42.74/0.2 +/- 0.52; Verbal: 8.2 +/- 4.94/18.05 +/- 4.63; Clock drawing: 2.07 +/- 2.56/6.03 +/- 11.25 for AD patients and control subjects, respectively). Mean scores for patients with AD and control subjects on allfour individual tests were significantly different (for each, p < 0.001). The mean time to complete the test for healthy control subjects was nine minutes and 18 seconds, for neuropsychological disorders nine minutes and six seconds, and for AD patients 13 minutes and 32 seconds (p < 0,001). Logistic regression analysis was used to determine the degree to which the battery discriminated between control subjects and patients with AD (sensitivity 92.73 percent and specificity 97.50 percent). We then separated the patients with MMSE > 20 and the same model of regression analysis was used. Sensitivity was 81.25 percent and specificity was 96.55 percent using 0.7 as the cutoff probability, and 93.75 and 96.55 percent, respectively, using 0.5 as the cutoffprobability. Neither age nor education and gender had an effect on the results. The Seven-Minute Screen appears highly sensitive to AD patients and may be useful in helping to make initial distinctions between patients with early dementia and normal elderly. PMID- 12083344 TI - Emotional response to social dancing and walks in persons with dementia. AB - The emotional response to social dancing and walks in persons with dementia was studied to better understand the feasibility, popularity, and meaning of these activities from the perspective of the patient. Social dance events and walks were videotaped and analyzed using Husserl's philosophy as a basis for the analysis. Six persons with dementia participated in the study. The results are described in terms of four interrelated themes: 1) the engaged body; 2) the caregivers' understanding, encouragement, and response to patients during the activity; 3) mutual tenderness and communion; and 4) environmental conditions. Results were then synthesized into a general assessment of the emotional states observed and reported in relation to the activities. PMID- 12083345 TI - Basic care I: the effect of dementia-specific training on certified nursing assistants and other staff. AB - This study evaluates the effectiveness of Basic Care I, a dementia-specific training course offered by the St. Louis Alzheimer's Association. Using three standardized measures, the effects of the course on knowledge gain, stress level, and sense of work-related self-esteem are examined Findings suggest that participation in Basic Care I increases retained learning, as 76.4 percent of the sample showed improvement on dementia knowledge scores. The outcomes of stress and self-esteem measures are inconclusive but indicate areas for future study. Implications for program planning are discussed. PMID- 12083346 TI - The psychological contract: enhancing productivity and its implications for long term care. AB - When hired, a new employee is usually given a job description and an explanation of benefits. In addition, the employee will also have a psychological contract with the organization. This contract, often unstated, reflects the main source of the employee's motivation to work hard. This is true of all groups of employees, including long-term care staff. Common examples of psychological contracts for long-term care administrative staff include autonomy, social acceptance, and being in the forefront of cutting-edge research. An awareness of these psychological contracts can result in better "fits" between employee aspirations and relevant long-term care organization tasks so that productivity is enhanced. This article outlines the steps necessary to create these good fits in ways that benefit both the organization and its employees. These recommendations are of particular relevance to administrators and supervisors in long-term carefacilities. PMID- 12083347 TI - Influence of aromatherapy on medication administration to residential-care residents with dementia and behavioral challenges. AB - Thirteen older persons (seven men and six women) in residential care participated as subjects in this study. All participants had histories of confusion due to dementia and were identified by staff as being consistently resistant to medication administration as indicated by vocal outbursts, moving away, or physical combativeness. Subjects were exposed to four aroma interventions during medication administration: 1) lavender vera (lavendula officinalis); 2) sweet orange (citrus aurantium); 3) tea tree (malaleuca alternifolia); and 4) no aroma (control). All medication administrations were videotaped for later data collection. Observers were trained to record frequency and duration of resistive behaviors during medication administration in allfour interventions for each subject. Reliability between two observers was extremely high. Results showed no statistically significant differences across all aroma conditions for either resistive behavior or duration of administration. Also, there were no statistically significant differences based on gender. This study indicates that aromatherapy does not reduce combative, resistive behaviors in individuals with dementia. Research with a larger sample in future studies may yield other results. PMID- 12083348 TI - Memories in the making: outcome-based evaluation of an art program for individuals with dementing illnesses. AB - The author presents the results of a pilot project, an outcomes-based evaluation of Memories in the Making: An art program for individuals with dementing illnesses. Sponsored by the Greater Cincinnati Chapter of the Alzheimer's Association and implemented at adult day and nursing home sites, participants in the weekly art program use paints to express themselves by creating colorful visual images on paper or fabric. The individuals experienced pleasurable, sustained activity while engaged in making art. Utilizing M.P. Lawton's conceptualization of psychological well-being as a framework, chapter staff developed an observational instrument, which measured objective and subjective indicators of the affect state and self-esteem of 41 participants in the art program. Overall, preliminary data suggests that participation in the weekly sessions contributed to the individuals' sense of well-being. However, more rigorous study is warranted using comparison groups to determine if indeed an enhanced sense of well-being can be attributed with certainty to the intervention of the Memories in the Making art program. PMID- 12083350 TI - Malaria vectors on Buka and Bougainville Islands, Papua New Guinea. AB - Anophelines were sampled from 82 locations on Buka and Bougainville islands in Papua New Guinea by larval collections, carbon dioxide-baited mosquito traps, and human biting catches. Anopheles farauti s.s. was collected in larval surveys but infrequently in mosquito traps on both islands; on Buka Island this species was readily collected in human biting catches. Anopheles farauti 2 was commonly collected in larval surveys on both islands; however, it was not collected in either mosquito traps or human biting catches. Anopheles punctulatus was found only on Buka Island, where it was commonly collected as larvae, but rarely in human biting catches and mosquito traps. Anopheles lungae was collected as larvae from only 1 site on Bougainville. Anopheles farauti s.s. fed consistently throughout the night (1900-0600 h); small peaks at midnight and dawn were not statistically significant. Of 1,156 An. farauti s.s. specimens examined by enzyme linked immunosorbent assay for malaria sporozoites, 20 were found to be positive; 12 were positive for Plasmodium falciparum and 8 were positive for P. vivax (247 variant = 5; 210 variant = 3). Anopheles farauti s.s. seems to be the major malaria vector on these islands, whereas An. punctulatus may play a minor role on Buka Island. Anophele jarauti 2 is unlikely to be involved in malaria transmission on Buka or Bougainville islands. PMID- 12083349 TI - Virtual Dementia Tour helps sensitize health care providers. AB - A review of the literature on sensitivity training among caregivers for the elderly revealed that no programs focused specifically on the cognitive changes that occur due to aging. Second Wind Dreams, a national nonprofit organization committed to improving society's perception of aging, conducted a study in which degenerative physical symptoms common for this population, such as impaired vision and motor skills, were simulated in a group of 146 subjects who worked in the field of elder care to give them a broader sense of the patient's perspective. Overwhelmingly, participants in the study came away with heightened awareness of the plight of confused elders and a strong sense that the high behavioral expectations caregivers have for dementia patients are unrealistic and need to change. PMID- 12083351 TI - Field evaluation of three plant-based insect repellents against malaria vectors in Vaca Diez Province, the Bolivian Amazon. AB - The efficacy of repellents against Anopheles darlingi, the main malaria vector in Bolivia, was evaluated. This mosquito has a peak in biting activity early in the evening. Three natural repellents (1 eucalyptus based, 1 neem based, and 1 containing several repellent essential oils) were tested in comparison with 15% deet in human landing catches in Bolivia. The eucalyptus-based repellent containing 30% p-menthane-diol applied at a dose similar to those used in practice gave 96.89% protection for 4 h. Deet gave 84.81% protection. The other 2 products did not provide significant protection from mosquito bites. PMID- 12083352 TI - Laboratory bioassay studies to determine methoprene susceptibility in a natural population of Ochlerotatus taeniorhynchus from the Florida keys. AB - Larvae of Florida Keys Ochlerotatus taeniorhynchus collected from No Name Key were colonized and evaluated against technical S-methoprene in laboratory beaker tests. Glassware was treated with a silanizing reagent before testing to prevent methoprene attachment to the glassware. The No Name Key strain was compared with a susceptible laboratory strain of Oc. taeniorhynchus. Five serial dilutions (0.0100, 0.0050, 0.0010, 0.0005, and 0.0001 microg/ml) and an untreated control were evaluated. Tests were conducted in water baths with a constant water temperature of 27+/-1 degrees C and 250 ml of 3 per thousand salt water. Twenty five late 3rd-stage larvae were placed in each beaker. Bioassay samples were analyzed by probit analysis and the median lethal concentration (LC50), 90% lethal concentration (LC90), and 95% lethal concentration (LC95) values; confidence limits; chi2 value; slope; and standard error were determined. The Florida Keys No Name Key strain exhibited no significant differences at the LC50, LC90, and LC95 levels from the laboratory strain in these studies. PMID- 12083353 TI - Influence of the lunar cycle on the activity of phlebotomine sand flies (Diptera: Psychodidae). AB - The influence of lunar phases on the activity of phlebotomine sand flies was evaluated in Vicosa, Minas Gerais, Brazil. The insects were collected by illuminated Shannon traps and Falcao light traps, between 1830 and 2230 h on 44 nights, divided between the dry and rainy seasons, and among each of the 4 lunar phases. A total of 888 sand flies was collected, belonging to 10 Lutzomyia species. The dominant species in both seasons of the year and in all lunar phases was Lutzomyia intermedia. A significant difference was found in the abundance of L. whitmani among lunar phases. No significant difference was found in frequency of sand flies collected among lunar phases. Females of L. intermedia initiated activity earlier during the crescent and full moon phases than during three quarter and new phases. Based on the premise that sand flies would exhibit normal phototaxis in the absence of moonlight, activity should be unaffected under a new moon, whereas light reflected by the moon in its brightest phases (crescent and full) should shift the period of activity of the sand flies so that it does not coincide with the period in which the moon is visible, or should reduce attractiveness of light traps to the insects by providing less background contrast. PMID- 12083354 TI - Comparison of two bioassay techniques for assessing the acute toxicity of pesticides to chironomid larvae (Diptera: Chironomidae). AB - Two container and substrate combinations were compared to determine which provided optimal survival of larvae of Chironomus tepperi and Glyptotendipes paripes in the absence of toxicants. Unfed final-stage larvae of G. paripes survived significantly (P < 0.05) better in waxed paper cups with sand substrate (92.8% after 3 days) than in glass tubes with a shredded paper substrate (85.3%). Survival of larvae of C. tepperi over the same period did not differ significantly in the 2 systems. Larvae of C. tepperi were bioassayed against 3 insecticides (technical and formulated imidacloprid, chlorpyrifos, and betacypermethrin) with both container and substrate combinations. Median lethal concentration values (24 h) obtained with waxed cups with sand were 1.8 times higher on average than those obtained with glass tubes with shredded paper (range 1.13-2.65 times). To determine the cause of this variability, solid-phase microextraction was used to measure changes in chlorpyrifos availability over time in the 2 bioassay systems. Chlorpyrifos concentrations in the waxed cups and sand system fell from 7.50 to 3.36 microg/liter over 24 h, probably as a consequence of chemical adsorption to the waxed surfaces. Chlorpyrifos concentrations in the glass tubes and paper system remained unchanged over this period. Excluding substrates from the containers had only a minor effect on chlorpyrifos availability. These results demonstrate that the behaviors of both test organisms and toxicants within bioassay systems need to be understood if the data generated with different systems are to be compared. Understanding how toxicant availability is affected by different container and substrate types is particularly important where bioassays are conducted with nominal concentration values rather than analytically determined exposure concentrations. PMID- 12083355 TI - A preliminary characterization of the physiological ecology of overwintering Anopheles mosquitoes in the midwestern USA. AB - Field-collected Anopheles quadrimaculatus and An. punctipennis adult mosquitoes from various types of overwintering hibernacula in southwestern Michigan and northern Indiana were examined for cold tolerance. Adult mosquitoes were collected on a monthly basis from October 1997 to April 1998. The mean supercooling point for adult for An. punctipennis was significantly lower than that for An. quadrimaculatus. The lower lethal temperatures for these species were -17 degrees C and -15 degrees C for An. punctipennis and An. quadrimaculatus, respectively. PMID- 12083357 TI - Discovery of Aedes albopictus infected with west nile virus in southeastern Pennsylvania. AB - In August 2000, Aedes albopictus was found in a CO2-baited Centers for Disease Control light trap in eastern Philadelphia, PA. In late September 2000, West Nile viral antigen was detected by reverse transcription-polymerase chain reaction testing from a pool of 2 Ae. albopictus mosquitoes that were collected in southwestern Montgomery County. PMID- 12083356 TI - An iridescent virus and a microsporidium in the biting midge Culicoides barbosai from Florida. AB - An iridescent virus and a microsporidium (Nosema sp.) were found infecting larvae of the biting midge Culicoides barbosai collected in Boynton Beach, Palm Beach County, Florida. Larvae were extracted from mud collected weekly during a 22 month period from March 1998 to December 1999. Virus-infected larvae were present in 4 collections made in March, April, and May 1998 and the average infection level was 4.7% (range 2.3-7.1%). The virus infected the fat body and produced an iridescent blue color in larvae because of the crystalline arrays of the particles. These virus particles were hexagonal, measured 99 nm from side to side, and had an electron-dense inner core. The site of infection of the Nosema sp. also was the fat body of the larva, which became opaque white. These opaque areas were the result of masses of oval binucleate spores measuring 2.7+/-0.1 x 4.4+/-0.3 microm (n = 25). The Nosema sp. was present in larvae of 14 collections made during the 22-month period, with an average infection level of 6.1% (range 0.3-21.4%). Although individual larvae infected with the iridescent virus and Nosema sp. occurred in 4 collections, no dual infections were observed. Larvae patently infected with the iridescent virus and Nosema sp. died before pupation. PMID- 12083358 TI - Mouthparts of male Aedes (Stegomyia) mosquitoes. AB - Mouthparts of adult males of 17 strains of 8 species from the subgenus Stegomyia of the genus Aedes, including 5 strains of Aedes aegypti and 6 strains of Aedes albopictus, were examined. Lengths of maxillae, mandibles, maxillary palpi, and proboscises were measured under light microscopy and their detailed structures were examined by scanning electron microscopy. Lengths were presented as ratios to proboscis lengths. In contrast to previous reports, mandibles were found in all 5 strains of male Ae. aegypti examined. Variations in maxillary and mandibular lengths were significant among strains, even within Ae. aegypti and Ae. albopictus. High variation of these structures among and within species indicates that the average length of these structures in only 1 species may not be a reliable representative of a subgenus, and those of 1 strain may not be reliable for a species. However, their range in length (maxillae 0.13-0.50, mandibles 0.04-0.17 length of the proboscis) may be regarded as a subgeneric attribute. Maxillae and mandibles distinctly shorter than the proboscis, together with their delicate structures and the large coefficient of variation, suggest that they exist only as vestigial structures. A positive correlation was found between lengths of maxillae and those of mandibles, but mandibles are usually shorter than maxillae. The hypopharynx is discernible from the labium wall by its texture and border, and this suggests that it was a free stylet in the past. PMID- 12083359 TI - Laboratory studies of Ochlerotatus samoanus in association with leaf axils of Freycinetia (Pandanaceae) in Samoa. AB - Three species of Freycinetia (Pandanaceae) (F. reineckei, F. storkii, and F. hombronii) were tested for oviposition preference of Ochlerotatus samoanus, a vector of filariasis in Samoa. Laboratory tests indicated that F. reineckei was preferred by this mosquito for oviposition. Eggs were preferentially deposited on a peeled or a moist dried leaf. The percentage hatch was highest when eggs were kept moist for longer than 4 days before submersion. Hatching was complete in less than 6 h. These studies allowed us to successfully rear Oc. samoanus in the laboratory, facilitating future studies on the biology and control of this important vector. PMID- 12083360 TI - Analysis of nectar and honeydew feeding in Aedes and Ochlerotatus mosquitoes. AB - Most research has investigated floral nectar as a source of carbohydrates for mosquitoes and has ignored homopteran honeydew. We have assessed the prevalence of honeydew and nectar feeding in 7 species of mosquitoes collected from Algonquin Provincial Park, Ontario, Canada. In total, 403 individuals were analyzed by thin-layer chromatography, with melezitose and stachyose as honeydew indicator sugars. From the 403 individuals, 214 contained sugars, of which only 8.8% had the honeydew-indicating sugars. We conclude that female Aedes and Ochlerotatus mosquitoes in Algonquin Provincial Park seem to feed infrequently on honeydew. PMID- 12083361 TI - Alcohol ingestion stimulates mosquito attraction. AB - Mosquito bites should be avoided because of the risk of contracting parasitic and viral diseases such as malaria, dengue fever, and several encephalitides. Although humans have been said to suffer more mosquito bites after ingesting liquor, little is known about whether that is true. Thirteen volunteers (12 men from 20 to 58 years old and a 24-year-old woman) were chosen as test hosts and a 30-year-old man was established as a control. We measured ethanol content in sweat, sweat production, and skin temperature before and after ingestion of 350 ml of beer (ethanol concentration 5.5%) by volunteers and compared them with a control subject. Our study demonstrated that percent mosquito landing on volunteers significantly increased after beer ingestion compared with before ingestion, showing clearly that drinking alcohol stimulates mosquito attraction. However, ethanol content in sweat and skin temperature did not show any correlation between alcohol ingestion and mosquito landings. This study shows that persons drinking alcohol should be careful about their increased risk to mosquito bites and therefore exposure to mosquito-borne diseases. PMID- 12083362 TI - Landing sites on the human body preferred by Aedes albopictus. AB - The landing sites on the human body preferred by Aedes albopictus were examined. Five male volunteers wearing only shorts stood in a mosquito net enclosure containing 120 proboscis-amputated Ae. albopitus. In separate tests, 9 male volunteers and 1 female volunteer lay supine during the test. The number of mosquitoes landing on each site of the volunteer's body was counted, and after completion of the test, his or her body temperature was recorded. When the subject was upright, the landing site most preferred by mosquitoes was the foot. When volunteers were supine, the foot also was the most preferred landing site, but the proportion of mosquitoes landing on the foot in this position in comparison with other sites was lower than when the Volunteer was in the upright position. The 2nd most preferred landing site was the hand, followed by the face. No correlation was found between preferred landing sites and body temperature. Factors other than temperature (e.g., human emanation) may influence mosquito behavior and landing site. PMID- 12083363 TI - N-cadherin involvement in the heterotypic adherence of malignant T-cells to epithelia. AB - N-cadherin, a cell adhesion molecule normally found in neural cell tissue, has been found recently to be expressed on the surface of malignant T-cells. The function of N-cadherin on these cells remains unclear. Heterotypic assays between Molt-3 T lymphoblastic leukemia cells and Caco-2 epithelial monolayers were examined under different conditions to assess the functional role of N-cadherin. The results indicate that adherence of Molt-3 cells to Caco-2 monolayers was reduced significantly following pretreatment of Molt-3 cells with 100 microM of an N-cadherin-derived antagonist decapeptide. In contrast, pretreatment of Molt-3 cells with an anti-N-cadherin antibody raised against the first 20 amino acids of N-cadherin sequence led to a surprisingly marked enhancement of Molt-3 cell adherence to Caco-2 monolayers. In addition, the presence of anti-N-cadherin antibody neutralized the inhibitory effect of anti-ICAM-1 on Molt-3 adhesion to Caco-2 monolayers. This novel finding demonstrates that external stimulus through the N-cadherin amino terminus can modulate adhesion of malignant T-cells to epithelia and may promote their ability to invade or metastasize to inflammatory sites. PMID- 12083364 TI - The carboxyl-terminal domains of MKP-1 and MKP-2 have inhibitory effects on their phosphatase activity. AB - Both the mitogen-activated protein kinase (MAPK) phosphatases MKP-1 and MKP-2 exert important feedback control of MAPK-mediated signaling events. The function of MKP-1 and MKP-2 is regulated via complex mechanisms, ranging from increased transcription of the MKP-1 and MKP-2 genes to post-translational catalytic activation of MKP-1 and MKP-2 proteins upon binding to their substrate MAPKs. In addition, MKP-1 stability increases upon ERK-dependent phosphorylation of two serine residues in its C-terminus. The C-terminal regions of MKP-1 and MKP-2, but not those of other MKPs, are homologous. To investigate the role of this domain, we have deleted the C-terminal tails from MKP-1 and MKP-2 and examined the effect of these deletions on their enzymatic activity. C-terminally truncated MKP-1 and MKP-2 exhibited, both in vivo and in vitro, substantially greater phosphatase activity towards their substrate MAPKs than did the full-length counterparts. However, C-terminal truncations did not significantly change either their substrate affinity, or their substrate-mediated catalytic activation. Basal phosphatase activity of the truncated proteins was also significantly higher than that of the wild-type counterparts. Collectively, these results suggest that the C-terminal domain may potentially play a role in the regulation of MKP-1 and MKP 2. PMID- 12083365 TI - Effects of chronic caloric restriction on mitochondrial respiration in the ischemic reperfused rat heart. AB - Dietary restriction increases life span and delays the development of age-related diseases in rodents. We have recently demonstrated that chronic dietary restriction is beneficial on recovery of heart function following ischemia. We studied whether the metabolic basis of this benefit is associated with alterations in mitochondrial respiration. Male Wistar rats were assigned to an ad libitum-fed (AL) group and a food restricted (FR) group, in which food intake was reduced to 55% of the amount consumed by the AL group. Following an 8-month period of restricted caloric intake, isolated working hearts perfused with glucose and high levels of fatty acids were subjected to global ischemia followed by reperfusion. At the end of reperfusion, total heart mitochondria respiration was assessed in the presence of pyruvate, tricarboxylic acid intermediates, and palmitoylcarnitine. Recovery of heart function following ischemia was greater in FR hearts compared to AL hearts. Paralleling these changes in heart function was an increase in state 3 respiration with pyruvate. The respiratory control ratios in the presence of pyruvate and tricarboxylic acid intermediates were higher in FR hearts compared to AL hearts, indicating well-coupled mitochondria. Overall energy production, expressed as the ADP:O ratio and the oxidative phosphorylation rate, was also improved in FR hearts. Our results indicate that the beneficial effect of FR on recovery of heart function following ischemia is associated with changes in mitochondrial respiration. PMID- 12083366 TI - NAD-dependent inhibition of the NAD-glycohydrolase activity in A549 cells. AB - NAD glycohydrolases are enzymes that catalyze the hydrolysis of NAD to produce ADP-ribose and nicotinamide. Regulation of these enzymes has not been fully elucidated. We have identified a NAD-glycohydrolase activity associated with the outer surface of the plasma membrane in human lung epithelial cell line A549. This activity is negatively regulated by its substrate beta-NAD but not by alpha NAD. Partial restoration of NADase activity after incubation of the cells with arginine or histidine, known ADP-ribose acceptors, suggests that inhibition be regulated by ADP-ribosylation. A549 do not undergo to apoptosis upon NAD treatment indicating that this effect be likely mediated by a cellular component(s) lacking in epithelial cells. PMID- 12083368 TI - Vanadium increases GLUT4 in diabetic rat skeletal muscle. AB - The effect of vanadium in lowering blood glucose in diabetic animals is well established; however, the exact mechanism of action of vanadium still eludes us. There are several reports from in vitro studies indicating that vanadium increases enzyme activity in insulin signalling pathways; however, these findings have not been duplicated in vivo. Glucose transporters (GLUT) have a major role to play in any glucoregulatory effects. Insulin dependent GLUT4 is a major glucose transporter present in skeletal muscle, adipocytes and heart. In the present study we found that the plasma glucose in streptozotocin (STZ) diabetic animals was restored to normal following treatment with a single dose of BMOV, an organic vanadium compound, given by oral gavage (0.6 mmol/kg), similar to the response with chronic BMOV treatment. The response to BMOV by oral gavage was rapid and the animals were normoglycemic within 24 h of treatment and still demonstrated a significant effect even after 72 h. Using a specific antibody against GLUT4 we found an overall reduction in the GLUT4 in the total membrane fraction in skeletal muscle of diabetic animals. However, with a single dose of BMOV the GLUT4 level was restored to normal. This is the first report that establishes a direct effect of vanadium on the regulation of GLUT4 expression in diabetic animals in vivo, and may at least partially explain the glucoregulatory effects of vanadium. PMID- 12083367 TI - Diazoxide protects the rabbit heart following cardioplegic ischemia. AB - K(ATP) channels are present in sarcolemmal and mitochondrial membranes. This study tests the hypothesis that opening mitochondrial K(ATP) channels with Diazoxide (DZ) improves tolerance to cardioplegic ischemia during surgery. Twenty two rabbit hearts were perfused with Krebs-Henseleit buffer (KHB) on a Langendorff apparatus and underwent 50 min of 37 degrees C global ischemia with St Thomas' cardioplegia (STCP). Hearts were divided into three groups. Ten (control) received no pretreatment. Seven (DZ) received 10 min of 30 microM DZ, a selective mitochondrial K(ATP) opener, in KHB before arrest with STCP containing 30 microM DZ. Five (5-HD + DZ) received 10 min of 100 microM sodium 5 hydroxydecanoate (5-HD), a selective mitochondrial K(ATP) channel blocker, followed by 10 min of 30 microM DZ and 100 microM 5-HD in KHB before arrest with STCP + 30 microM DZ + 100 microM 5-HD. LV developed pressure (LVDP), dP/dt and coronary flow (CF) were measured after 60 min of reperfusion. Diazoxide pretreatment significantly improved the recovery of LV function and coronary flow compared to control (LVDP: 49 +/- 5* vs. 31 +/- 4; +dP/dtmax 927 +/- 93 vs. 507 +/- 85 mmHg/sec*; CF 33 +/- 4 vs. 22 +/- 2 ml/min, *p < 0.05). Mitochondria K(ATP) channel blockade with 5-HD prevented DZ's salutary effect on the recovery of LV and vascular function. Diazoxide pretreatment protects the rabbit heart during cardioplegic ischemia by opening mitochondrial K(ATP) channels. Opening mitochondrial K(ATP) channels may be a new strategy for improving myocardial protection during cardiac surgery. PMID- 12083369 TI - Effect of myocardial stunning on thiol status, myofibrillar ATPase and troponin I proteolysis. AB - To investigate the mechanism underlying postischemic contractile dysfunction (myocardial stunning) we examined myocardial sulflhydryl group content, myofibrillar Ca2+-dependent Mg2+-ATPase activity and protein profile after global ischemia and reperfusion. The Langerdorff-perfused rabbit hearts were subjected to 15 min normothermic ischemia followed by 10 min reperfusion and myofibrils were isolated from homogenates of left ventricular tissues. Depressed contractile function during reperfusion was accompanied by a decrease in total sulfhydryl group content. However, myofibrillar protein profile was unchanged and Western immunoblotting analysis showed no significant differences in troponin I immunoreactive bands between control and stunned hearts. Likewise, myofibrillar Mg2+-ATPase activity was unaltered after ischemia and reperfusion. We conclude that myocardial stunning is not caused by altered myofibrillar function and protein degradation but may be partly due to the oxidative modification of as yet undefined proteins. PMID- 12083370 TI - Cholestin inhibits cholesterol synthesis and secretion in hepatic cells (HepG2). AB - Hyperlipidemia is a well-known risk factor for atherosclerosis and statins are widely used to treat patients with elevated levels of lipids in their plasma. Notwithstanding the proven benefits of statin drugs on both primary and secondary prevention of heart disease, the high cost of statin treatment, in addition to possible side effects such as liver function abnormalities, may limit their widespread use. We conducted a study on a natural product as an alternative to statin treatment. Cholestin, a dietary supplement, is prepared from rice fermented with red yeast (Monascus purpureus), which has been shown to significantly decrease total cholesterol levels in hyperlipidemic subjects. Our objective was to determine the cellular effect of Cholestin on cholesterol synthesis in human hepatic cells (HepG2) and the mechanism by which it caused a change in lipid metabolism. Cholestin had a direct inhibitory effect on HMG-CoA reductase activity (78-69% of control). Cholesterol levels in HepG2 cells treated with Cholestin (25-100 microg/mL) were significantly reduced in a dose-dependent manner (81-45% of control, respectively). This reduction was associated with decreased synthesis and secretion of both unesterified cholesterol (54-31 and 33 14% of control, respectively) and cholesteryl ester (18-6 and 37-19% of control, respectively). These results indicate that one of the anti-hyperlipidemic actions of Cholestin is a consequence of an inhibitory effect on cholesterol biosynthesis in hepatic cells and provide the first documentation of a biomolecular action of red yeast rice. PMID- 12083371 TI - Elongation factor 2 as a target for selective inhibition of protein synthesis in vitro by the novel aromatic bisamidine. AB - The effect of the novel aromatic bisamidine 1 on protein synthesis in cell-free translational system isolated from rat livers was studied. The bisamidine 1 caused inhibition of [14C]leucine incorporation into proteins proportionally to its concentration. To establish a precise mechanism of inhibition, we evaluated the effect of the bisamidine 1 on the isolated ribosomes and purified to homogeneity elongation factors. Preincubation of the bisamidine 1 with ribosomes resulted in partial inhibition of their activity in whole elongation system. The eucaryotic elongation factor 1 (eEF-1) was not significantly affected by the bisamidine 1. In contrast to eEF-1, the bisamidine 1 preincubated with the eucaryotic elongation factor 2 (eEF-2) caused total inhibition of its activity in the translocation process. The inhibitory effect of the bisamidine 1 on eEF-2 activity was confirmed in diphtheria toxin-dependent ADP-ribosylation reaction. The results suggest a high action specificity of the bisamidine 1 as potential anticancer drug, since the primary target seems to be highly conserved protein elongation factor 2. PMID- 12083373 TI - Sex differences in p-aminohippuric acid transport in rat kidney: role of membrane fluidity and expression of OAT1. AB - Sex differences in the kinetic parameters of p-aminohippuric acid (PAH) transport in brush border (BBMV) and basolateral (BLMV) membrane vesicles from kidney cortex have been observed. Membrane fluidity of BBMV was higher in females as compared with male rats as indicated by anisotropy values (0.1897 +/- 0.0010 vs. 0.2003 +/- 0.0014, p < 0.05, for females and males respectively). Membrane fluidity of BLMV were similar in both sexes. Western blot studies revealed that OAT1 protein in female BLMV was present at only 40% of level found in BLMV from male rats. The lower expression of OAT1 in BLMV in association with the higher BBMV fluidity (which may affect the affinity of PAH transporter in this membrane domain) observed in females may be responsible, at least in part, for the gender difference described in renal PAH secretion. PMID- 12083372 TI - Expression of atrial natriuretic peptide receptor-A antagonizes the mitogen activated protein kinases (Erk2 and P38MAPK) in cultured human vascular smooth muscle cells. AB - To understand the signaling mechanisms of atrial natriuretic peptide (ANP) receptor-A (NPRA), we studied the effect of the ANP/NPRA system on mitogen activated protein kinases (MAPKs), with particular emphasis on the extracellular regulated kinase (Erk2) and stress-activated protein kinase (p38MAPK) in cultured human vascular smooth muscle cells (HVSMC). Angiotensin II (ANG II) and platelet derived growth factor (PDGF) stimulated the immunoreactive Erk2 and p38MAPK activities and their protein levels by 2-4 fold. The pretreatment of cells with ANP significantly inhibited the agonist-stimulated Erk2 and p38MAPK activities and protein expression by 65-75% in HVSMC transiently transfected with NPRA, as compared with only 18-22% inhibition in vector-transfected cells. The pretreatment of cells with KT5823, an inhibitor of cGMP-dependent protein kinase (PKG), reversed the inhibitory effects of ANP on MAPK activities and protein expression by 90-95%. PD98059, which inhibits Erk2 by directly inhibiting the MAPK-kinase (MEK), and SB202192, a selective antagonist of p38MAPK, blocked the Erk2 and p38MAPK activities, respectively. Interestingly, ANP stimulated the MAPK phosphatase-3 (MKP-3) protein levels by more than 3-fold in HVSMC over-expressing NPRA, suggesting that ANP-dependent inhibition of MAPKs may also proceed by stimulating the phosphatase cascade. These present findings provide the evidence that ANP exerts inhibitory effects on agonist-stimulated MAPKs (Erk2 and p38MAPK) activities and protein levels in a 2-fold manner: by antagonizing the up-stream signaling pathways and by activation of MKP-3 to counter-regulate MAPKs in a cGMP and PKG-dependent manner. Our results identify a signal transduction pathway in HVSMC that could contribute to vascular remodeling and structural changes in human hypertension. PMID- 12083374 TI - HnRNP-A1 binds directly to double-stranded DNA in vitro within a 36 bp sequence. AB - The heterogeneous nuclear ribonucleoprotein A1 (hnRNP-A1) is known as an RNA- and single-stranded DNA-binding protein involved in alternative splicing of mRNA, RNA transport and maintenance of chromosome telomere length. In this study we tested whether this protein could bind directly to double-stranded DNA (dsDNA). Using PCR amplification of target DNA-sequences from human chromosome 11q13 followed by their incubation with hnRNP-A1 and atomic force microscopy (AFM) of the DNA/protein complexes, we found that this protein bound to DNA within a 36 bp sequence. These results were confirmed by electrophoretic mobility shift assay (EMSA). This sequence was found widely dispersed throughout the genome. There is no overlap between the 36 bp sequence and known target sequences in RNA for binding binRNP-A1. PMID- 12083375 TI - Selection of a WEHI-3B leukemia cell subclone resistant to inhibition by cholera toxin. AB - The studies on the inhibitory effect exerted by Cholera Toxin (CT) on cell growth and proliferation indicate a remarkable heterogeneity of cell response suggesting that the inhibition represents the final event of many different ways or mechanisms. After CT binding, cAMP accumulation may not occur (as in L1210 leukemia cells) or, when occurring (as in SR-4987 stromal cells), may not be coupled with the antiproliferative effect of CT. In WEHI-3B cells CT binds a Gal GalNac-GM1b receptor and the anticlonogenic effect of CT seems correlated with cAMP accumulation. To demonstrate the central role of cAMP in WEHI-3B cells, starting from the sensitive cell strain we selected and established a clone of WEHI-3B resistant to CT. This revertant clone (WEHI-3B/CT/REV) is currently cultured in the absence of CT and in the proliferation assay shows a dramatic resistance (>46,000 than the parental cells). Stimulation ofWEHI-3B/CT/REV cells by cholera toxin failed to enhance cAMP and the ganglioside-CT binding studied on Thin Layer Chromatography (TLC) blots showed that the resistant cells lost the spot correspondent to the migration of Gal-GalNac-GM1b ganglioside. Both the lines respond at the same level to the adenylate cyclase stimulation by forskolin and the incorporation of GM1a did not decrease the resistance of WEHl-3B/CT/REV. These data confirm that Gal-GalNac-GM1b is the most important functional receptor for CT in WEHI-3B cells able to transduce the signal by enhancing cAMP which in turn inhibits cell proliferation (probably by cAMP dependent protein kinase activation). Our study describes the first cell line resistant to CT originated from a susceptible parental strain and provides a new interesting cell model for studying the cAMP dependent mechanisms involved in cell growth regulation. PMID- 12083376 TI - Oltipraz protects the passive smoke induced changes in renal glyoxalase system of rats. AB - The present study was performed to investigate the effect of oltipraz on passive smoke-induced alteration in renal glyoxalase system of rats. Adult Sprague-Dawley rats were exposed daily to passive cigarette smoke in a whole-body exposure chamber 6 h per day for 2, 4 and 12 weeks. The animals being sacrificed after 2 and 12 weeks were maintained on control diet, powdered 4% Teklad rat chow (Harlan Teklad, Madison, WI, USA). The 4 weeks group was divided into three subgroups, one receiving control diet, other two receiving control diet supplemented with two doses of oltipraz (either 167 or 500 ppm), starting 1 week prior to initiation of smoke exposure until the end of the experiment. The activity of glyoxalase I was higher in animals exposed for 4 and 12 weeks of passive smoke than those exposed for 2 weeks. There was no significant difference between 4 and 12 weeks. Glyoxalase II activity was lower in animals exposed to passive smoke for 4 weeks than those exposed for 2 weeks. However, the activity approached the basal level after 12 weeks of exposure. Furthermore, oltipraz treatment maintained the activity of both glyoxalase closer to the basal levels. PMID- 12083377 TI - Cloning of the 5'-flanking region of the murine bone morphogenetic protein-7 gene. AB - BMP-7, a member of the bone morphogenetic protein subfamily of the TGFbeta superfamily is highly expressed in the murine kidney. BMP-7 is involved in fetal nephron development and mesenchymal to epithelial cell differentiation. Constitutive BMP-7 expression is found in tubular and glomerular epithelial cells of the adult kidney. BMP-7 may play a role in physiology and pathophysiology of the adult kidney since BMP-7 gene expression in acute renal ischemia is diminished and injection of recombinant BMP-7 into rats with ischemic acute renal failure preserves renal function. In order to investigate the transcriptional regulation of BMP-7, this study was undertaken to clone and characterize the promoter of the murine BMP-7 gene. A 1394 bp sequence of the 5'-flanking region of the BMP-7 gene was isolated and subcloned. No TATA and CAAT box consensus motifs could be identified as shown for promoters of other BMPs. Using in vitro transfection assays, the 5'-flanking region revealed moderate to strong basal promoter activity. PMA increased basal BMP-7 promoter activity. Thus BMP-7 gene transcription might involve at least in part a PKC-dependent pathway. The cloning of a 5'-flanking region of the BMP-7 gene should provide a useful tool for future studies on the transcriptional regulation of BMP-7 gene expression. PMID- 12083378 TI - Change of apocytochrome c translocation across membrane in consequence of hydrophobic segment deletion. AB - Wild-type apocytochrome c and its hydrophobic segment deleted mutants, named delta28-39, delta72-86 and delta28-29/72-86 were constructed, expressed and highly purified respectively. Insertion ability into phospholipid monolayer, inducing leakage of entrapped fluorescent dye fluorescein sulfonate (FS) from liposomes, and translocation across model membrane system showed that the wild type apoprotein and delta28-39 almost exhibited the same characteristics, while mutants with segment 72-86 deletion did not. Furthermore, CD spectra, intrinsic fluorescence emission spectra, and the accessibility of the protein to the fluorescence quenchers: KI, acrylamide and HB demonstrated that the segment 72-86 deletion has a significant effect on the conformational changes of apocytochrome c following its interaction with phospholipid. On the basis of these results it is postulated that the C-terminal hydrophobic segment 72-86 plays an important role in the translocation of apocytochrome c across membrane. PMID- 12083379 TI - K-FGF mediated transformation and induction of metastatic potential involves altered ornithine decarboxylase and S-adenosylmethionine decarboxylase expression -role in cellular invasion. AB - Omithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMDC) expression was investigated in NIH-3T3 fibroblasts that secrete K-FGF. Correlations between altered ODC and SAMDC expression and malignant potential were determined. Increased ODC and SAMDC expression was associated with increased expression of both ODC and SAMDC mRNA and enzyme activity levels. Transcriptional and post-transcriptional regulatory mechanisms were found to account for the increased expression of both ODC and SAMDC. Amplification of the ODC gene also played a role. Correlations between the expression of ODC and the invasion ability of the K-FGF overexpressing cells were also found. Additionally, putrescine, which is a cellular polyamine, was found to play a role in determining the nature of the invasive capacity of the K-FGF overexpressing cells. The results of this study which established correlations between alterations in the expression of ODC and SAMDC, the key rate limiting and regulatory activities in the synthesis of cellular polyamines, and malignant potential as a consequence of K-FGF overexpression supports a model which suggests that growth factor modulation of ODC and SAMDC expression is part of the altered growth regulatory program associated with cellular transformation and malignant progression. PMID- 12083380 TI - Nitration/S-nitrosation of proteins by peroxynitrite-treatment and subsequent modification by glutathione S-transferase and glutathione peroxidase. AB - In various peroxynitrite (PN)-treated proteins, the formations of stable 3 nitrotyrosine (nitration) and labile S-nitrosocysteine (S-nitrosation) were observed by employing rapid Western blot in 6 h. The steps of SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and membrane-blotting were performed at 4 degrees C. It was noted that the intensity of immunoreactive bands specific for anti nitrotyrosine was stronger than that specific for anti-S-nitrosocysteine. Additionally, the intensity was in the manner of a dose-dependency of PN. Nitration/S-nitrosation were formed in the following treated proteins, including bovine serum albumin (BSA), DNase-1, ceruloplasmin, catalase and hemoglobin (Hb). The incubation of PN-pretreated hemoglobin with 1 mM reduced glutathione (GSH) did not change immunoreactivity significantly. However, the addition of glutathione S-transferase (GST) or glutathione peroxidase (GPX) to the above incubation mixture, resulted in decreased immunoreactivity, suggesting GSH may form a transition complex with PN-pretreated hemoglobin and/or partially reduce/modify the treated hemoglobin, thereby increasing the accessibility for the subsequent modification by GST or GPX. Such decreased immunoreactivity indicates that nitrotyrosine and S-nitrosocysteine of treated hemoglobin was, indeed, further modified via (a) converting -NO2 to -NH2 in tyrosine residues, (b) denitrating -NO2 directly/indirectly in tyrosine residues, and/or (c) changing -S-NO to -SH in cysteine residues, or denitrosation. The findings imply similar enzymatic modifications of proteins may also occur in vivo, and therefore play a pivotal role in the NO-related cellular signaling cascade(s). PMID- 12083381 TI - Hypertrophy of cultured adult rat ventricular cardiomyocytes induced by antibodies against the insulin-like growth factor (IGF)-I or the IGF-I receptor is IGF-II-dependent. AB - Antibodies against the insulin-like growth factor-I (IGF-I) or the IGF-I receptor (IGF-IR) directly initiate a rapid (within 6 h) hypertrophy of isolated adult rat ventricular cardiomyocytes cultured in the absence of serum. Further, cardiomyocytes treated with either of these agonistic antibodies upregulate the expression of their genes for insulin-like growth factor-II (IGF-II) and the IGF II receptor (IGF-IIR). Genistein, an inhibitor of the tyrosine kinase IGF-IR, also induces the cardiomyocytes to hypertrophy. Anti-IGF-II antibody inhibits the cardiomyocyte hypertrophy induced by anti-IGF-I and anti-IGF-IR antibodies or by genistein. Results are consistent with a model in which local production of IGF II is upregulated when the IGF-IR signaling pathway is blocked and in which an IGF-II-mediated pathway, likely involving the IGF-IIR, then stimulates hypertrophy of the cardiomyocytes. PMID- 12083382 TI - Effects of a lichen galactomannan and its vanadyl (IV) complex on peritoneal macrophages and leishmanicidal activity. AB - A galactomannan (GMPOLY) isolated from lichen Ramalina celastri was complexed with vanadyl ion (IV;VO) forming the complex GMPOLY-VO. Both GMPOLY and GMPOLY-VO diminished the superoxide anion production by macrophages triggered with PMA, the complex giving rise to this effect at concentrations 100 times lower than GMPOLY. Macrophages treated with GMPOLY enhanced the nitric oxide production (40%), this effect not being observed when interferon-gamma (IFN-gamma) or IFN-gamma plus lipopolysaccharide (LPS) were present. No effect on nitric oxide production was observed by treatment of macrophage with GMPOLY-VO. Both GMPOLY and GMPOLY-VO exhibited leishmanicidal effects on the amastigote form of Leishmania amazonesis, but only GMPOLY-VO inhibited the growth of promastigote form. PMID- 12083383 TI - 4-Aminopyridine: effects on electrical activity during ischemia and reperfusion in perfused rat hearts. AB - We investigated the effects of 2 and 4 mM 4-aminopyridine (4-AP,--blocker of the transient outward current I(to) on the electrophysiological response to regional ischemia and reperfusion. Spontaneously beating rat hearts were subjected to coronary occlusion (10 min) followed by reperfusion. The surface electrogram and the membrane potential from subepicardial left ventricular cells were recorded throughout. The basal effect of 4-AP was a dose dependent increase in the action potential duration (APD90) without changes in the resting potential or the heart rate. During early ischemia resting depolarization (from 87.4 +/- 1.9-70.1 +/- 2.5 mV in the controls) was enhanced by 4 mM, 4-AP (84.3 +/- 1.4 mV vs. 61.7 +/- 1.3 mV) whereas APD90 increased by 73.5%. These effects resulted in a marked reduction in the duration of diastolic intervals that led to conduction failure and aborted responses. A partial recovery was found by the end of ischemia concomitant with APD90 shortening in both, control and 4-AP treated hearts. On reperfusion, 4-AP did not influence the initial incidence of ventricular tachyarrhythmias but decreased their duration from 531.5 +/- 56.3-260.7 +/- 100 sec (2 mM) and to 75.6 +/- 10.5 sec (4 mM). These data confirm others obtained by Henry et al. in isolated cells indicating that ischemia induces sequential changes in several K+ conductances. In addition, they show that changes in action potential characteristics may exert beneficial effects on reperfusion arrhythmias by acting on the arrhythmic substrate without suppressing the trigger mechanism. PMID- 12083384 TI - Monocyte-macrophage differentiation in vitro: modulation by extracellular matrix protein substratum. AB - The influence of extracellular matrix (ECM) on monocyte-macrophage (mo-mphi) differentiation was investigated using an in vitro model with human peripheral blood mononuclear cells (PBMC) maintained on different matrix protein substrata. Macrophage specific markers associated with differentiation studied were, (a) endocytosis of modified proteins; (b) appearance of mphi specific matrix metalloproteinases (MMPs); (c) activities of myeloperoxidase (MPO) and beta-D glucuronidase; (d) changes in the expression of cell surface antigens. As the duration of monocytes in culture increased, a progressive increase in the rate of differentiation was seen as evidenced by mphi specific functions such as endocytosis of 125[I] acetyl BSA and the appearance of gelatinases A and B. Significantly higher rate of endocytosis and production of MMPs were found in monocytes maintained on fibronectin (FN) and COL I than on COL IV (FN > COL I > COL IV) indicating that cells in contact with stromal components differentiate at a faster rate. FACS analysis done on cells maintained in vitro for phenotypic profile characteristic to mo-mphi differentiation showed downregulation of CD14 occurring in a substratum dependent manner viz, (FN > COL IV > COL I) and upregulation of CD71 was high in cells maintained on COL I and COL IV. Intracellular enzymatic activities such as MPO significantly decreased irrespective of matrix substrata, while beta-D-glucuronidase activity increased in a substratum dependent manner (FN > COL I > COL IV). Pretreatment of cells with genistein significantly decreased the secretion of MMPs, particularly MMP 9 in cells maintained on ECM protein (FN) indicating a phosphorylation dependent signalling process in mediating matrix effect. The results of these in vitro studies on mphi specific markers suggest that apart from the diffusible factors, the microenvironment as provided by various matrix proteins particularly FN can modulate mo-mphi differentiation. PMID- 12083385 TI - The effect of hyposmotic and isosmotic cell swelling on the intracellular [Ca2+] in lactating rat mammary acinar cells. AB - The effect of hyposmotic and isosmotic cell swelling on the free intracellular calcium concentration ([Ca2+]i) in rat mammary acinar cells has been examined using the fura-2 dye technique. Ahyposmotic shock (40% reduction) increased the [Ca2+]i in rat mammary acinar cells in a fashion which was transient; the [Ca2+]i returned to a value similar to that found under isomotic conditions within 180 sec. The increase in the [Ca2+]i was dependent upon the extent of the osmotic shock. The hyposmotically-activated increase in the [Ca2+]i could not be attributed to a reduction in extracellular Na+ or a change in the ionic strength of the incubation medium. Thapsigargin (1 microM) enhanced the hyposmotically activated increase in the [Ca2+]i. Isosmotic swelling of rat mammary acinar cells, using urea, had no significant effect on the [Ca2+]i. Similarly, a hyperosmotic shock did not affect the [Ca2+]i in rat mammary acinar cells. It appears that the effect of cell swelling on the [Ca2+]i in rat mammary acinar cells depends on how the cells are swollen (hyposmotic vs. isosmotic). This finding may have important physiological implications given that it is predicted that mammary cell volume will change in vivo under isomotic conditions. PMID- 12083387 TI - Dye purity and dye standardization for biological staining. AB - This review starts with a short discussion of what is meant by a pure dye and a brief description of how dyes are produced. A listing of the types of impurities encountered in dyes is made, followed by technical investigations and assessments of dyes and their impurities including methods for separating, identifying and assaying dye components. In the second part of the review, descriptions are given of the standardized staining method approach using standard staining methods for assessing stains, and practical responses to stain impurity including commercial quality control, third-party quality control and standardization of reagents, protocols and documentation. Finally, reference is made to the current state of affairs in the dye field. PMID- 12083386 TI - Purification of a lipid-activated and Ca2+-independent protein kinase from the mantle tissue of Mytilus galloprovincialis Lmk. AB - A phospholipid-sensitive Ca2+-independent protein kinase (p105) was purified to homogeneity from mantle tissue of the mussel Mytilus galloprovincialis Lmk., employing consecutively DE-52 cellulose, Sephacryl S-200 and Biogel HTP chromatographies. The purified enzyme appeared as a single band on 10% SDS-PAGE, and had a molecular weight of 105 kDa. The positive Western blotting of the purified eluate for anti-human-PKCdelta and PKCepsilon suggests that the enzyme from mussel mantle may be an ancestral nPKC isoform, with the kinetic properties of the enzyme very close to those of PKCepsilon isoform of vertebrates. Western blotting of samples from different steps of purification using specific mouse anti-p105, showed two protein bands in samples from the initial steps. However, only one band was detected in the Biogel-HTP eluate, the most purified fraction. The purification steps did not affect the presence of P-serine in p105. No P tyrosine peptides were detected in any of the purification steps. These results open a new field of work on the study of several molecular processes related to energetic metabolism and reproduction in molluscs, whose regulation is associated with the activation of protein kinases. PMID- 12083388 TI - An improved staining method for intervertebral disc tissue. AB - The objective of this study was to design a new staining procedure for human disc tissue for visualizing both collagen and proteoglycan-matrix components on the same histology section. Weigert's hematoxylin, alcian blue and picrosirius red were combined to produce distinctive staining of collagen (red), proteoglycans (blue) and cellular elements of the intervertebral disc. This novel stain reveals sharp details of collagen composition in the perilacunar, territorial and intraterritorial extracellular matrix, and concomitantly demonstrates the presence of proteoglycan accumulations around cells in the lacunar spaces and in the extracellular matrix. These details reveal variations within the tissue that would not be apparent with routine stains. PMID- 12083389 TI - Immunohistochemical localization of c-mos at the light and electron microscope level in non-small cell lung carcinomas. AB - C-mos is a cytoplasmic upstream activator of the mitogen-activating protein kinase pathway with serine-threonine kinase activity. It plays a well established and vital role in oocyte maturation by participating in metaphase II arrest and meiotic asymmetric division, but little is known about its function in somatic cells. Recently, we observed overexpressed c-mos in a portion of non-small cell lung carcinomas (NSCLCS). In particular, c-mos immunoreactivity was detected in tumor cell nuclei in addition to its expected cytoplasmic localization, and c-mos overexpression was associated with chromosomal instability among other findings. To verify our earlier observations and to clarify further the role of c-mos in NSCLCS, we examined its distribution by both light and electron microscopy. We detected c-mos in the cytoplasm and/or nucleus of a portion of tumor cells and fibroblasts. In particular, granular immunoreactivity was observed in the cytoplasm closely associated with the rough endoplasmic reticulum. Nuclear staining was confirmed and was often found near the nuclear membrane, as well as in some large multilobular, possibly aneuploid, nuclei. C-mos positivity was also found in the nuclei of tumor cells undergoing apoptosis. Furthermore, c-mos was detected in areas with diminished vascularization. It should be noted that nuclear staining was found at the ultrastructural level more extensively than at the light microscope study. This suggests a masking effect by the hematoxylin nuclear counterstain. PMID- 12083390 TI - Improved demonstration of mast cells using alcian blue tetrakis (methylpyridium) chloride. AB - Many batches of alcian blue dye are incompletely soluble at the low pH used for demonstrating mast cells. An improved technique using alcian blue tetrakis (methylpyridium) chloride (alcian blue pyridine variant) is described here. It produces stronger mast cell staining than other alcian blue stains tested. PMID- 12083391 TI - MAP-2 immunolabeling can distinguish diffuse from dense-core amyloid plaques in brains with Alzheimer's disease. AB - Alzheimer's disease (AD) neuropathology is characterized by the presence of diffuse and dense-core (neuritic) amyloid plaques in specific areas of the brain. The origin of these plaques and the relationship between them is poorly understood. Current methods to identify clearly these types of plaques in the AD brains are largely dependent upon morphological characteristics. Dense-core amyloid plaques in the entorhinal cortex and hippocampus of AD brains might arise from the lysis of neurons overburdened by excessive intracellular deposition of amyloid beta1-42 (Abeta42) peptide. The local release of active lysosomal enzymes, which persist within these plaques, might degrade most of the released intracellular proteins, leaving behind only those that are resistant to proteolytic digestion, such as ubiquitin, tau, neurofilament proteins and amyloid. To test the possibility that proteins that are sensitive to proteolysis may be degraded selectively in plaques, we used immunohistochemistry to examine the distribution of microtubule-associated protein-2 (MAP-2), a protein localized primarily in neuronal dendrites and known to be sensitive to proteolysis. Uniform MAP-2 immunolabeling was detected throughout the somatodendritic compartment of neurons in age-matched control cortical brain tissues as well as throughout areas of Abeta42-positive diffuse plaques in AD brains. In contrast, analysis of serial sections revealed that MAP-2 was absent from Abeta42-positive dense-core plaques in AD brains. Our results indicate that this differential MAP-2 immunolabeling pattern among plaques may be employed as a reliable and sensitive method to distinguish dense-core plaques from diffuse plaques within AD brain tissue. Furthermore, this biochemical distinction indicates that dense-core and diffuse plaques are formed by different mechanisms. PMID- 12083392 TI - The challenge of using SNPs in the understanding and treatment of disease. PMID- 12083393 TI - The SNP consortium: summary of a private consortium effort to develop an applied map of the human genome. PMID- 12083394 TI - Primer design for PCR and sequencing in high-throughput analysis of SNPs. AB - To achieve high-throughput analysis of allele frequencies in human SNPs, we have developed automated methodsfor designing PCR and DNA sequencing primers. We found we could run the PCR assays at quite stringent, uniform conditions. The design process used freely available databases, including dbSNP, SNPper, and TSC, and publicly available software including RepeatMasker and Primer3. We describe parameters for the software and other considerations that increase experimental success. As anticipated. some assays filed at the design stage due primarily to the genomic locations of repetitive sequences, extreme GC content regions, or lack of sufficient sequence. However, over 23,000 assays, including 96% of those recently analyzed, have been experimentally successfuL Similar design methods could be usedfor PCR assays in any organism with substantial available sequence. PMID- 12083395 TI - Characterization of cytochrome P450 2D6 alleles using the Invader system. AB - The cytochrome p450 (CYP) superfamily comprises enzymes that play an essential role in the transformation of medically relevant compounds. Accurate genotyping of polymorphisms in members of this family is drawing increasing interest because certain allelic variants may result in either loss of efficacy or toxic accumulation of therapeutic agents. Debrisoquine 4-hydroxylase, or CYP2D6, is among the most widely studied of the CYPs. The complexity of the CYP2D6 genomic region, including pseudogenes, gene deletions, and gene duplications, has offered numerous challenges to developing a genotyping strategy. We describe a comprehensive CYP2D6 genotyping strategy that employs both a PCR/Invader genotyping assay system and an Invader genomic copy number assay The Invader system is a homogeneous, isothermal, highly specific, and robust signal amplification system. Resultsfrom II CYP2D6 assays in an alle frequency study compare well to published allele frequency values for Caucasians. Further, Invader assays provided unambiguous genotyping determinations for 100% of the 171 samples that yielded a visible PCR product on an agarose gel. A copy number assay yielded only one equivocal result in 205 samples. We identified 17 single-copy individuals and 17 three-copy (or more) individuals. PMID- 12083396 TI - Introduction to SNPs: discovery of markers for disease. PMID- 12083397 TI - TempliPhi, phi29 DNA polymerase based rolling circle amplification of templates for DNA sequencing. AB - We have developed a novel, isothermal DNA amplification strategy that employs phi29 DNA polymerase and rolling circle amplification to generate high-quality templates for DNA sequencing reactions. The TempliPhi DNA amplification kits take advantage of the fact that cloned DNA is typically obtained in circular vectors, which are readily replicated in vitro using phi29 DNA polymerase by a rolling circle mechanism. This single subunit, proofreading DNA polymerase has excellent processivity and strand displacement properties for generation of multiple, tandem double-stranded copies of the circular DNA, generating as much as 10(7) fold amplification. Large amounts of product (1-3 microg) can be obtained in as little as 4 hours. Input DNA can be as little as 0.01 ng of purified plasmid DNA, a single bacterial colony, or a 1 microL of a saturated overnight culture. Additionally, the presence of an associated proof reading function within the phi29 DNA polymerase ensures high-fidelity amplification. Once completed, the product DNA can be used directly in sequencing reactions. Additionally, the properties of phi29 DNA polymerase and its use in applications such as amplification ofhuman genomic DNA for genotyping studies is discussed. PMID- 12083398 TI - New generation pharmacogenomic tools: a SNP linkage disequilibrium Map, validated SNP assay resource, and high-throughput instrumentation system for large-scale genetic studies. AB - Since public and private efforts announced the first draft of the human genome last year, researchers have reported great numbers of single nucleotide polymorphisms (SNPs). We believe that the availability of well-mapped, quality SNP markers constitutes the gateway to a revolution in genetics and personalized medicine that will lead to better diagnosis and treatment of common complex disorders. A new generation of tools and public SNP resources for pharmacogenomic and genetic studies--specifically for candidate-gene, candidate-region, and whole genome association studies--will form part of the new scientific landscape. This will only be possible through the greater accessibility of SNP resources and superior high-throughput instrumentation-assay systems that enable affordable, highly productive large-scale genetic studies. We are contributing to this effort by developing a high-quality linkage disequilibrium SNP marker map and an accompanying set of ready-to-use, validated SNP assays across every gene in the human genome. This effort incorporates both the public sequence and SNP data sources, and Celera Genomics' human genome assembly and enormous resource ofphysically mapped SNPs (approximately 4,000,000 unique records). This article discusses our approach and methodology for designing the map, choosing quality SNPs, designing and validating these assays, and obtaining population frequency ofthe polymorphisms. We also discuss an advanced, high-performance SNP assay chemisty--a new generation of the TaqMan probe-based, 5' nuclease assay-and high throughput instrumentation-software system for large-scale genotyping. We provide the new SNP map and validation information, validated SNP assays and reagents, and instrumentation systems as a novel resource for genetic discoveries. PMID- 12083399 TI - BeadArray technology: enabling an accurate, cost-effective approach to high throughput genotyping. AB - The Human Genome Project has opened the door to personalized medicine, provided that human genetic diversity can be analyzed in a high-throughput and cost effective way Illumina has developed a genotyping system that combines very high throughput and accuracy with low cost per SNP analysis. The system uses our BeadArray platform, a high level of multiplexing, and modular, scalable automation to meet the requirements for cost-effective, genome-wide linkage disequilibrium studies. As implemented in a high-throughput genotyping service facility at Illumina, the system has a current capacity of one million SNP assays per day and is easily expandable. Each SNP call is associated with a quality score that correlates with accuracy PMID- 12083400 TI - A strategy for the rapid discovery of disease markers using the MassARRAY system. PMID- 12083401 TI - SNPstream UHT: ultra-high throughput SNP genotyping for pharmacogenomics and drug discovery. AB - Single nucleotide polymorphism (SNP) genotyping is playing an increasing role in genome mapping, pharmacogenetic studies, and drug discovery. To date, genome-wide scans and studies involving thousands of SNPs and samples have been hampered by the lack of a system that can perform genotyping with cost-effective throughput, accuracy, and reliability. To address this need, Orrhid has developed an automated, ultra-high throughput system, SNPstream UHT, which uses multiplexed PCR in conjunction with our next generation SNP-IT tag array single base extension genotyping technology The system employs oligonucleotide microarrays manufactured in a 384-well format on a novel glass-bottomed plate. Multiplexed PCR and genotyping are performed in homogeneous reactions, and assay results are read by direct two-color fluorescence on the SNPstream UHTArray Imager. The systems flexibility enables large projects involving thousands of SNPs and thousands of samples as well as small projects that have hundreds of SNPs and hundreds of samples to be done cost effectively. We have successfully demonstrated this system in greater than 1,000,000 genotyping assays with >96% of samples giving genotypes with >99% accuracy PMID- 12083402 TI - SNP market view: opportunities, technologies, and products. PMID- 12083403 TI - Marine fat and brain function in manic-depressive psychosis and schizophrenia: circumstantial evidence for a 2nd aquatic period. PMID- 12083404 TI - Some aspects of the multifaceted dependence of early humanity on water. PMID- 12083405 TI - The fetus and newborn considered as water-babies. PMID- 12083407 TI - The continental shelf hypothesis. PMID- 12083406 TI - Was man more aquatic in the past. What happens when you change the paradigm? PMID- 12083408 TI - Cerebral evolution. AB - We suggest that the evolution of the large human brain could not have occurred on the savannahs of Africa. It depended on avoiding the trap of universal loss of brain capacity amongst all land species as they evolved larger bodies. The nutrition at the water's front would have given evolving humans the edge over other hominid species. They would have had access to a rich, plentiful and easily harvested food resource, rich in DHA, trace elements and anti-oxidants, for little energy expenditure. This does not deny the possibility that the men hunted for meat. However, a reliable and consistent nutrient dense food resource, rich in DHA would have been especially important for women during pregnancy and lactation and of course the children during the period of fetal and neonatal brain development and reproductive maturation. PMID- 12083409 TI - Introduction: The Saugstad Prize for Elaine Morgan. The conditions of human evolution. PMID- 12083410 TI - Retinoids: essential nutrients for the central nervous system. PMID- 12083411 TI - Human nature is unique in the mismatch between the usual diet and the need for "food for the brain" (marine fat, DHA). Adding marine fat is beneficial in schizophrenia and manic-depressive psychosis. This underlines brain dysfunction in these neurological disorders is associated with deficient intake of marine fat(DHA). PMID- 12083412 TI - The effect of marine oils on inflammatory reactions in humans. PMID- 12083413 TI - Prevention of fatal cardiac arrhythmias by polyunsaturated fatty acids. PMID- 12083414 TI - Is marine fat anti-epileptogenic? PMID- 12083415 TI - Marine fat and human health. Introduction. PMID- 12083416 TI - Human umbilical vein endothelial cells express complement receptor 1 (CD35) and complement receptor 4 (CD11c/CD18) in vitro. AB - We have examined complement receptors on human umbilical vein endothelial cells (HUVEC) and found that they express complement receptor 1 (CR1, CD35) and complement receptor 4 (CR4, CD11c/CD18), but not complement receptor 3 (CR3, CD11b/CD18). Binding of monoclonal antibodies against CR1 (CD35) and CR4 (CD11c/CD18) to HUVEC was demonstrated by flow cytometry. The presence of the corresponding mRNAs was confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequencing of the amplified cDNA fragments. When HUVEC were treated with inflammatory mediators, chemotactic agents or the secretagogue phorbol-12-myristate-13-acetate (PMA), no change in reactivity to CR1 (CD35) or CR4 (CD11c/CD18) monoclonal antibodies was detected on the surface of the cells compared with untreated cells. The presence of CR1 (CD35) and CR4 (CD1c/CD18) on HUVEC indicates that endothelial cells (EC) have the potential to bind C3b and iC3b, respectively, which both mediate biological effects in the course of complement activation. PMID- 12083417 TI - Functional effects of eotaxin are selectively upregulated on IL-5 transgenic mouse eosinophils. AB - Synergistic interactions between cytokines, chemokines and adhesion molecules may facilitate the selective recruitment of eosinophils into sites of allergic inflammation. Ovalbumin-sensitized IL5TG mice responded to antigen challenge with robust airway eosinophilia 24 and 72 hr post-exposure. Adhesion molecule expression and functional responsiveness of immune cells derived from IL5TG mice to various inflammatory mediators were evaluated. IL5TG-derived eosinophils, but not neutrophils, expressed higher levels of CD49d and CD11b relative to WT. Functional responsiveness to eotaxin was increased in IL5TG eosinophils as demonstrated by a 10x increase in its potency in producing actin polymerization and 3x increase in CD11b upregulation relative to WT. These data are consistent with increased CCR3 expression on IL5TG eosinophils. Responsiveness of eosinophils to LTB4 or MIP-1alpha was similar between WT and IL-5TG mice. These data provide evidence of synergy between eosinophil-specific cytokines and chemokines that may promote accumulation of this cell type under conditions of allergic inflammation in vivo. PMID- 12083418 TI - Peroxisome proliferator-activated receptor agonists inhibit inflammatory edema and hyperalgesia. AB - Previous studies have produced conflicting data on the contribution of the peroxisome proliferator-activated receptors (PPARs) to the inflammatory process. This study investigated the effects of several PPARalpha and PPARgamma subtype specific agonists on the inflammation and hyperalgesia produced by intraplantar carrageenan injection in unanesthetized male Sprague-Dawley rats. Intraperitoneal administration of PPARalpha agonists reduced edema in parallel to their potencies determined in vitro. Perfluorooctanoic acid (PFOA) inhibited carrageenan-induced edema in a dose-dependent manner, and also reduced thermal hypersensitivity. Furthermore, PFOA produced much more robust effects when administered 0.5-24 hrs before carrageenan, as compared to when it was administered 1.5 hrs after carrageenan. Intraperitoneal administration of similar doses of the PPARgamma agonist rosiglitazone, but not the less potent agonist, troglitazone, reduced edema when administered before but not after carrageenan. We conclude that systemic administration of potent PPARalpha and PPARgamma agonists exert anti hyperalgesic and/or antiinflammatory actions in vivo, possibly by interfering with the initiation of inflammation. PMID- 12083420 TI - Do NSAIDs affect the progression of osteoarthritis? AB - NSAIDs are widely used to alleviate the symptoms of OA. It remains controversial as to what effects these agents have on the progression of OA. In vitro studies showed several types of NSAIDs (e.g., sodium salicylate, indomethacin) inhibited the synthesis of cartilage matrix component, but some types of NSAIDs (e.g., aceclofenac, meloxicam, nimesulide) increased the matrix component synthesis and protected the chondrocytes against apoptosis, while others (e.g., piroxicam) had no effects. Studies in animal models verified that NSAIDs had favourable or detrimental action on OA progression, even the same NSAID (e.g., naproxen, tiaprofenic acid) had reverse effects on articular cartilage in different studies. Preliminary clinical trials revealed some NSAIDs such as indomethacin had a negative influence on joint structure, other NSAIDs such as diclofenac and naproxen had no acceleration of radiographic damage to OA within 2-years of treatment. So far, there are no convincing data to show the widely used NSAIDs and recommended selective COX-2 inhibitor have favourable effects on cartilage. Therefore, it is necessary and valuable to clarify the effects of these NSAIDs on cartilage in patients with OA using validated non-invasive methods such as MRI. PMID- 12083419 TI - Temporal and mechanistic effects of heat shock on LPS-mediated degradation of IkappaBalpha in macrophages. AB - Previous studies demonstrated important interactions between the heat shock response and the IkappaBalpha/NF-kappaB pathway when these two pathways are induced sequentially. One such interaction involves the ability of heat shock to inhibit subsequent degradation of IkappaBalpha in response to a proinflammatory signal. Herein we investigated the temporal relationship between recovery from heat shock and inhibition of IkappaBalpha degradation, and the proximal mechanisms by which heat shock inhibits degradation of IkappaBalpha in macrophages. In RAW 264.7 murine macrophages, prior heat shock inhibited LPS mediated IkappaBalpha degradation up to 4 h after recovery from heat shock, and this effect correlated with inhibition of LPS-mediated activation of NF-kappaB. Beyond these recovery periods, heat shock did not inhibit IkappaBalpha degradation. IkappaB kinase (IKK) assays demonstrated that heat shock inhibited LPS-mediated activation of IKK up to 1 h after recovery from heat shock. Heat shock also increased intracellular phosphatase activity, and inhibition of intracellular phosphatase activity partially reversed the ability of heat shock to inhibit both LPS-mediated degradation of IkappaBalpha and LPS-mediated activation of IKK. These data demonstrate that the ability of heat shock to inhibit degradation of IkappaBalpha is dependent on the recovery period between the heat shock stimulus and the proinflammatory stimulus. The mechanism by which heat shock inhibits degradation of IkappaBalpha involves dual modulation of IKK and intracellular phosphatase activity. PMID- 12083422 TI - An improved resazurin-based cytotoxicity assay for hepatic cells. AB - A simple resazurin-based cytotoxicity assay is presented for screening of cytotoxicity in hepatocytes and liver cell lines. Human hepatoma (HepG2) cells in 96-well culture plates were exposed to known toxic (cisplatin, 5-fluorouracil, ethionine, flufenamic acid, and diflunisal) and control (transplatin, 5 chlorouracil, methionine, and acetylsalicylic acid) compounds for 1-3 days, and resazurin (5 micromol/L) was added. A conventional short-term (1 h) assay was first performed, where cytotoxicity is indicated by decreased reduction of resazurin to its fluorescent product resorufin. Our improved assay consists of additionally measuring fluorescence 2-4 days later, when cytotoxicity is indicated by a striking increase in the concentration of resorufin, resulting from two distinct processes. First, viable liver-derived cells slowly convert resorufin to nonfluorescent metabolites. Fluorescence of control cell wells decreased to background during a 2- to 4-day exposure to resazurin. This metabolism of resorufin was largely blocked by dicumarol and to lesser extents by disulfiram and SKF525a. Second, dead or dying cells slowly convert resazurin to resorufin but do not further metabolize resorufin; thus this fluorescent metabolite accumulates to high levels in wells with dead cells by 2 to 4 days. A similar increase in fluorescence associated with cytotoxicity was observed in primary cultures of rat hepatocytes using the long-term resazurin-based assay. In addition to an improved signal relative to the short-term assay, the inversion of the fluorescent signal from high = alive short-term to high = dead long-term allows determination of two independent cytotoxicity endpoints after addition of one innocuous vital dye. PMID- 12083421 TI - Investigation of parenchymal cell differentiation in organotypic slice culture of mouse fetal liver under administration of sodium butyrate. AB - The fetal mouse liver tissues in our organotypic slice culture were spread and flattened for at least 3 weeks; small, round cells were distributed in the center and polygonal cells were seen in the periphery. Ultrastructurally, polygonal cells showed abundant rough endoplasmic reticulum and mitochondria. They expressed albumin (ALB) and alpha-fetoprotein (AFP) for at least 3 weeks, and Cx32-immunoreactivity was also seen in a plaque on the cells. Many proliferating cell nuclear antigen (PCNA)-positive cells were observed at the periphery, and there were scattered CK-19-positive cells. The spreading of the fetal liver tissue in organotypic slice culture was reduced in medium containing sodium butyrate (SB). The expression of ALB was well maintained in polyglonal cells of the SB(+) group 3 weeks after culture and AFP-immunoreactivity was decreased in the SB(+) group. The concentration of ALB in the medium was significantly higher in the SB(+) than in the SB(-) group. CK-19-positive cells in the SB(+) group were increased in number more than those in the SB(-) group. PCNA-positive cells were less numerous in the SB(+) group, and Cx32-positive plaques were increased. SB can help immature hepatocytes to differentiate into the mature type and the cholangiocytic lineage, reducing their proliferation. These findings suggest that parenchymal cells in our organotypic slice culture of the fetal mouse liver can maintain structure and function as in vivo for the long term, and SB is shown to be a differentiation inducer of parenchymal cells in the slice culture. PMID- 12083423 TI - Suppression of sulfur mustard-increased IL-8 in human keratinocyte cell cultures by serine protease inhibitors: implications for toxicity and medical countermeasures. AB - The toxicity of the chemical warfare blistering agent sulfur mustard (2,2' dichlorodiethyl sulfide; SM) has been investigated for nearly a century; however, the toxicological mechanisms of SM remain obscure and no antidote exists. The similarity of dermal-epidermal separation caused by SM exposure, proteolysis, and certain bullous diseases has fostered the hypothesis that SM vesication involves proteolysis and/or inflammation. Compound screening conducted by the US Army Medical Research Institute of Chemical Defense established that topical application of three tested serine protease inhibitors could reduce SM toxicity in the mouse ear vesicant model. Although most of the drugs with efficacy for SM toxicity in rodent models are anti-inflammatory compounds, no in vitro assay is in current use for screening of potential anti-inflammatory SM antidotes. IL-8 is a potent neutrophil chemotactic cytokine that is increased in human epidermal keratinocyte (HEK) cell cultures following exposure to SM and has been proposed as a marker for SM-induced inflammation. This study was conducted to establish in vitro screening of IL-8 in SM-exposed HEK as a possible model for evaluating candidate compounds prior to in vivo testing. We chose two protease inhibitors, one from those shown as successful in the MEVM (ethyl p-guanidinobenzoate hydrochloride, ICD 1579) and a prototypic inhibitor of trypsin, N-tosyl-L-lysine chloromethyl ketone (TLCK). TLCK (62.5 to 1000 micromol/L) or ICD 1579 (31.25 to 1000 micromol/L) was added to HEK cell cultures 1 h after SM exposure (200 micromol/L) and dose-dependently suppressed SM-increased IL-8. The suppression of SM-increased IL-8 by a class of drug candidate compounds such as protease inhibitors may provide a mechanistic marker that helps predict future medical countermeasures for SM toxicity and reduces the need for testing in animal models. PMID- 12083424 TI - Bilirubin induces loss of membrane lipids and exposure of phosphatidylserine in human erythrocytes. AB - Unconjugated bilirubin increasingly binds to erythrocytes as the bilirubin-to albumin molar ratio exceeds unity, leading to toxic manifestations that can culminate in cell lysis. Our previous studies showed that bilirubin induces the release of lipids from erythrocyte membranes. In the present work, those studies were extended in order to characterize the alterations of membrane lipid composition and evaluate whether bilirubin leads to a loss of phospholipid asymmetry. To this end, human erythrocytes were incubated with several bilirubin to-albumin molar ratios (0.5 to 5), and cholesterol as well as the total and the individual classes of phospholipids were determined. To detect erythrocytes with phosphatidylserine at the outer surface, the number of annexin V-positive cells was determined following incubation with bilirubin, fixing its molar ratio to albumin at 3. The results demonstrate profound changes in erythrocyte membrane composition, including modified cholesterol and phospholipid content. The release of membrane cholesterol, as well as of total and individual classes of phospholipids at molar ratios > or = 1, indicates that damage of erythrocytes may occur in severely ill jaundiced neonates. The loss of the inner-located phospholipids, phosphatidylethanolamine and phosphatidylserine, points to a redistribution of phospholipids in the membrane bilayer. This was confirmed by the exposure of phosphatidylserine at the outer cell surface. In conclusion, this study demonstrates that bilirubin induces loss of membrane lipids and externalization of phosphatidylserine in human erythrocytes. These features may facilitate hemolysis and erythrophagocytosis, thus contributing to enhanced bilirubin production and anemia during severe neonatal hyperbilirubinemia. PMID- 12083425 TI - Isolation of pig colonic crypts for cytotoxic assay of luminal compounds: effects of hydrogen sulfide, ammonia, and deoxycholic acid. AB - Some colonic luminal molecules resulting from bacterial metabolism of alimentary or endogenous compounds are believed to exert various effects on the colonic epithelial cell physiology. We isolated surface epithelial cells and intact colonic crypts in order to test bacterial metabolites in the pig model, which is often considered relevant for extrapolation to the physiopathology of the human gastrointestinal tract. Using colonocytes isolated with EDTA, we found that the initial cell viability, estimated by the membrane integrity and oxidative capacity measurement, fell rapidly despite several experimental attempts to preserve it such as the use of a medium designed to increase the adherence of epithelial cells and of a coated extracellular matrix, the presence in the culture medium of the oxidative substrate butyrate, and the use of an inhibitor of the caspases involved in cell apoptosis. In contrast, using dispase and collagenase as proteolytic agents, we were able to obtain pig colonic crypts that maintain an excellent membrane integrity after 4 h. Using this preparation, we were able to test the presumably cytotoxic luminal compounds hydrogen sulfide, ammonia, and deoxycholic acid on colonic crypt viability. Of these, only deoxycholic acid was found to significantly alter the cellular membrane integrity. It is concluded that pig colonic crypts can be useful for the in vitro appraisal of the cytotoxic properties of luminal compounds. PMID- 12083426 TI - Culturing precision-cut human prostate slices as an in vitro model of prostate pathobiology. AB - Due to the complex morphology of the prostate, it was hypothesized that precision cut tissue slices from human prostate would provide a unique in vitro model. Precision-cut slices were generated from zones of human prostate and their viability was assessed under conditions of different media for up to 120 h. Slices were also exposed to several concentrations of CdCI2, which was used as a model toxicant. Maintenance of both stromal and epithelial cells was noted; however, there was a gradual loss of luminal epithelial cells when the medium was not supplemented with dihydrotestosterone (DHT). Minimal leakage of lactate dehydrogenase occurred throughout the incubation. Prostate-specific antigen (PSA) was detected in the medium at all time points, although the rates of secretion fell over time. There was a loss of PSA-positive cells when the medium was not supplemented with DHT, consistent with a loss of luminal cells, whereas PSA positive cells were maintained in the DHT-supplemented media. A proliferation of basal cells was observed in the presence of media containing 10% fetal bovine serum. Exposure of slices to CdCl2 demonstrated a dose-response effect ranging from proliferation to complete cellular necrosis. Given the retention of stromal epithelial interactions and the use of acquired human tissue, prostate slices represent a unique in vitro model for investigating human prostate pathobiology. PMID- 12083427 TI - Phase specific approaches to the epidemiology and prevention of sexually transmitted diseases. PMID- 12083428 TI - A pilot study of a rapid assessment method to identify places for AIDS prevention in Cape Town, South Africa. AB - A multidisciplinary approach appears promising for focusing interventions. The PLACE method (priorities for local AIDS control efforts) identifies where prevention programmes can access sexual networks containing individuals with high rates of new partner acquisition and provides information about whether sexually transmitted disease (STD) or AIDS prevention programmes are in place at those sites. The method provides useful information at relatively low cost, regardless of the phase of the HIV or STD epidemic. For example, in the growth phase of an epidemic, information on where people with high rates of new partner acquisition meet new sexual partners could be used to inform targeted surveillance or screening programmes to catch early cases. PMID- 12083429 TI - Response of a sexually transmitted infection epidemic to a treatment and prevention programme in Nairobi, Kenya. AB - Although it seems possible in a developing country context such as Kenya, given appropriate inputs and a sound approach, to shift a sexually transmitted disease (STI) epidemic from phase II to III, it is not entirely clear how to go beyond this stage, to low levels of endemicity or even elimination. Perhaps the most important challenge now is to expand STI treatment and community STI/HIV prevention programmes to a much larger scale. Although successful programmes have been implemented in many areas of sub-Saharan Africa on a small scale, a significant impact in reducing the STI/HIV burden will not occur until programme reach is expanded to district, provincial, and national levels. PMID- 12083430 TI - STI epidemics in the Indian Ocean region: can the phase be assessed? AB - The Indian Ocean provides a unique opportunity to curb the HIV epidemic in its nascent phase through strengthening STI control programmes. Making effective and appropriate health services available should be regarded as the first priority for STI control in the region and, whenever possible, core groups should be identified and targeted to interrupt transmission within such networks. PMID- 12083431 TI - Phase specific strategies for the prevention, control, and elimination of sexually transmitted diseases: background country profile, England and Wales. AB - The development of non-invasive tests for evidence of current or past infection offers new prospects for carrying out population studies of sexually transmitted infection (STI) epidemiology, and for improved understanding of the behavioural and biological determinants of STI transmission. In turn this may guide screening, treatment, and behavioural interventions. The evidence base for many interventions is sparse and randomised trials of interventions are in their early days. Increasingly, linked behavioural and biological outcomes will be required to measure effectiveness. Integrated efforts are needed at a national level to achieve greater coordination between STD, HIV, and contraception services, school education, and broader public education and programmes. PMID- 12083432 TI - Phase specific strategies for the prevention, control, and elimination of sexually transmitted infections: case study in Lambeth, Southwark, and Lewisham, London, UK. AB - Lambeth, Southwark, and Lewisham are three London boroughs that probably have the worst set of indicators of sexual health in the United Kingdom. This area is used to illustrate an urban epidemic of bacterial sexually transmitted infections in an industrialised country with well developed public health infrastructure, and the generalisability of the phase specific model for prevention and control of sexually trasmitted infections. The results show that national strategies for preventing and controlling sexually transmitted infections comprise a limited repertoire of activities that fit with those suggested by the phase specific model. Specific strategies for men and for young black heterosexuals in the most heavily affected areas should be developed. There is, however, insufficient empirical evidence about whether particular interventions introduced at different epidemic stages are effective. PMID- 12083433 TI - Geographical variations in the epidemiology of bacterial sexually transmitted infections in Manitoba, Canada. AB - Feasible epidemiological approaches are required to make a better assessment of the stage of an epidemic and to monitor its transition through various phases. Application of the Lorenz curve and Gini coefficient to summarise the inequality in STD incidence rates between jurisdictions in Manitoba, Canada, was found to provide useful insights into the concentration of these epidemics over time and thus their transition through epidemic phases. Further exploration of the statistical properties of these and other indices of inequality and their potential application to STD epidemiology is warranted. New epidemiological tools are also required for better monitoring of the impact of prevention and control activities and to inform the content of these activities. PMID- 12083434 TI - The geographical and temporal evolution of sexually transmitted disease epidemics. AB - In developing appropriate interventions to control sexually transmitted infections (STIs) it is important to understand the distribution of infections, and how this distribution is influenced by the dynamic nature of epidemics. The epidemiology of STIs has been described as a series of phases, which loosely positions a population within the history of an epidemic. The relation between a mathematical theory describing STI epidemiology and the taxonomy of "phases" is explored in this review of the transmission dynamics of STIs. The likelihood of spread and persistence of infection depends upon the basic reproductive number, whereas the endemic prevalence of infection depends upon the way in which heterogeneity in risk focuses the density dependent constraints limiting the spread of infection. All interventions aimed at reducing the prevalence and incidence of infection must act through the reproductive number. The key parameters determining the reproductive number and the relevance of risk behaviours depend upon the biology of the particular infection. The division between high transmission probability, short duration infections and low transmission probability, long duration infections is explored. PMID- 12083435 TI - Gonorrhoea and chlamydia core groups and sexual networks in Manitoba. AB - This paper summarises the results of the R0 equation in sexually transmitted infection (STI) repeaters in Manitoba, Canada, in the early 1990s, with both concurrent and more recent descriptions of sexual networks in the same population. The research presented provides empirical network and sex partner data to refine definitions of sexual networks and core groups in phase IV epidemics. New challenges for both practice and research are also discussed. PMID- 12083437 TI - Risk network structure in the early epidemic phase of HIV transmission in Colorado Springs. AB - This study describes the risk network structure of persons with HIV infection during its early epidemic phase in Colorado Springs, USA, using analysis of community-wide HIV/AIDS contact tracing records (sexual and injecting drug partners) from 1985 to 1999. Paired partner information from other STD/HIV programme records was used to augment network connections. Analyses were conducted with and without this supplemental information. The results suggest that a combined dendritic and cyclic structural network pattern is associated with low to moderate HIV propagation in Colorado Springs, and may account for the absence of intense propagation of the virus. PMID- 12083436 TI - Sexual network structure as an indicator of epidemic phase. AB - Ascertaining epidemic phase for a sexually transmitted disease (STD) has depended on secular trend data which often contain significant artefacts. The usefulness of sexual network structure as an indicator of STD epidemic phase is explored in an analysis of community wide genital chlamydia reports, with network analysis of interviewed cases and linked sexual partners, in Colorado Springs, USA, 1996 to 1999. In this period, the chlamydia case rate per 100,000 increased by 46%. Three quarters of cases (n=4953) were interviewed, nominating 7365 partners; these, combined with index cases, made up the 9114 persons in the network. Epidemiologic analysis of cases suggests that secular trend increases are artefactual. Network analysis supports this view: overall network structure is fragmented and dendritic, notably lacking the cyclic (closed loops) structures associated with network cohesion and thus with efficient STD transmission. Comparison of network structure with that of an intense STD outbreak (characterised by numerous cyclic structures) suggests low level or declining endemic rather than epidemic chlamydia transmission during the study interval. These observations accord with intuitive and stochastic predictions. PMID- 12083438 TI - Heterosexual outbreak of infectious syphilis: epidemiological and ethnographic analysis and implications for control. AB - This study describes the epidemiology and ethnography of an outbreak of infectious syphilis in Vancouver, British Columbia. Between 1996 and 1999, British Columbias's rate of infectious syphilis rose from 0.5 to 3.4 per 100,000, with a dense concentration of cases among sex trade workers, their clients, and street-involved people in the downtown eastside area of Vancouver. Sexual networks were imported cases with secondary spread (dyads and triads), large densely connected dendritic networks of sex trade workers and clients, or occasional starburst networks among gay men. Only 232 of 429 partners were documented as having been treated (54% of those named, or 0.9 per case). The geographical and demographic concentration of this outbreak led to consideration of a programme of focused mass treatment with single dose azithromycin. PMID- 12083439 TI - Risky sexual behaviours among injection drugs users with high HIV prevalence: implications for STD control. AB - Injection drug use is inextricably linked to commercial sex work and the transmission of sexually transmitted disease (STD). In many communities prevention efforts have been stalled owing to the marginal existence of this community. This study describes the sexual activities, condom use, reported STDs, and commercial sex work in a large cohort of injection drug users. Seventy two per cent of male and 92% of female subjects in the cohort were sexually active. Among female subjects, 57% reported more than 100 lifetime partners. Condoms were generally not used with regular partners, used about half the time with casual partners, and used about 80% of the time with paying partners. Female sex workers were more likely to have unstable housing and to report incarceration in the previous six months. Reducing the transmission of STDs and HIV in drug using communities is a public health priority. While existing prevention programmes should be strengthened, innovative approaches to STD surveillance, diagnosis, and prevention are needed. PMID- 12083440 TI - From community to clients: the professionalisation of HIV prevention among gay men and its implications for intervention selection. AB - Forces at work are described which encourage professionalisation and a reliance on one on one HIV prevention interventions among gay men. Community involvement is intrinsically linked to epidemic phases; when the threat diminishes, so does the community's ability to sustain community level interventions. The area of structural and environmental interventions, which can reinforce safe behaviour when community interest in collective action wanes, provides a potential complementary solution for prevention workers, researchers, and funders alike. PMID- 12083441 TI - Populations, pathogens, and epidemic phases: closing the gap between theory and practice in the prevention of sexually transmitted diseases. AB - The phase specific model for the prevention and control of sexually transmitted infections (STI) offers new insights into the strategic planning of programmes. The model illustrates the importance of modifying the focus of prevention and control activities to different subpopulations as the epidemic evolves over time. However, the practical application of phase specific approaches will depend on an understanding of the variability and determinants in the trajectory by which STI epidemics progress through epidemic phases. This paper draws on empirical observations from diverse populations to explore the influence of sexual behaviour patterns in populations, the biological characteristics of STI pathogens, and the population-pathogen interactions in relation to epidemic trajectories. In addition, various approaches to the determination of epidemic phase are presented. PMID- 12083442 TI - Canadian STI national goals and phase specific strategies. AB - National goals should be "SMART"-specific, measurable, achievable, resource sensitive, and timed. To be meaningful, these goals must be sustainable and realistic within the time frame and fiscal limitations. Multisector partnerships are essential for the goals to become operational. PMID- 12083443 TI - The impact of the phase of an epidemic of sexually transmitted infection on the evolution of the organism. AB - The evolution of any sexually transmitted organism will be influenced by prevailing epidemiological interactions. The optimum strategy for an organism to overcome treatment, either through drug resistance or cryptic infections, depends upon whether the method for identifying patients is passive (treating symptoms alone favours asymptomatic organisms) or active (screening favours resistant organisms). The use of mathematical models of competing strains of infection allows theoretical predictions for the outcome of evolution under a range of assumptions about potential phenotypes. The course of pathogen evolution has implications for the success of interventions, but the predictions presented need to be tested at the level of the community in carefully monitored interventions. PMID- 12083444 TI - Determinants of STD epidemics: implications for phase appropriate intervention strategies. AB - Determinants of evolving epidemics of sexually transmitted diseases (STD) are equally influenced by the evolution of the STD epidemics themselves and by the evolution of human societies. A temporal approach to STD transmission dynamics suggests the need to monitor infectivity, rate of exposure between infected and susceptible individuals, and duration of infectiousness in societies. Different indicators may be used to monitor rate of exposure in the general population and in core groups. In addition, underlying determinants of STD epidemics such as poverty, inequality, racial/ethnic discrimination, unemployment, sex ratio, volume of migration, and health care coverage and quality are important variables to monitor through a surveillance system focused on social context. Ongoing large scale societal changes including urbanisation, globalisation, increasing inequality, and increasing volume of migrant populations may affect the evolution of STD epidemics. Globalised STD epidemics could pose a major challenge to local public health systems. PMID- 12083445 TI - Diverse realities: sexually transmitted infections and HIV in India. AB - There are many features that make India a vulnerable country as far as a sexually transmitted infection (STI)/HIV epidemic is concerned. These include the lack of a strong evidence base on which to formulate decision making, a pluralistic and often unregulated health sector, and a highly vulnerable population. Nonetheless, India has shown strong commitment to other areas of a comprehensive reproductive health care programme, and may be able to do so in the field of STI/HIV control. Vast numbers of people in India are severely disadvantaged in terms of income, education, power structures, and gender. Addressing these basic issues of human rights lies at the core of achieving better health outcomes. PMID- 12083446 TI - The profile and context of the epidemics of sexually transmitted infections including HIV in Zimbabwe. AB - Zimbabwe has widespread and widely disseminated epidemics of most major sexually transmitted infections (STI) including HIV. This epidemiological situation is examined from a broad historical perspective, exploring the interactions between the population incidence of STI and the social profile of the country. The results suggest opportunities for upstream prevention efforts. Examples of these include: integration of prevention with care and social support; increasing general communication and openness about sexuality; economic support initiatives including income generating and micro-credit programmes, offering living accommodations for families in cities, mines, and military camps; and programmes focusing on adolescents before they become sexually active. PMID- 12083447 TI - Sex in the city: sexual behaviour, societal change, and STDs in Saigon. AB - HIV infection is increasing among sex workers and injection drug users in southern Vietnam. Vietnamese sex workers returning from Cambodia are an important factor. This phase I growth stage is being accelerated by widespread prostitution and escalating heroin use. Sexually transmitted disease (STD) rates are significant in sex workers but low in the general population. STD epidemics in developing countries may not follow the dynamic topology that is common in developed countries. Vietnam has the potential for significant HIV and STD epidemics but also the capacity to respond to these threats. PMID- 12083448 TI - The effect of HIV, behavioural change, and STD syndromic management on STD epidemiology in sub-Saharan Africa: simulations of Uganda. AB - An assessment was made of how the HIV epidemic may have influenced sexually transmitted disease (STD) epidemiology in Uganda, and how HIV would affect the effectiveness of syndromic STD treatment programmes during different stages of the epidemic. The dynamic transmission model STDSIM was used to simulate the spread of HIV and four bacterial and one viral STD. Model parameters were quantified using demographic, behavioural, and epidemiological data from rural Rakai and other Ugandan populations. The findings suggest that severe HIV epidemics can markedly alter STD epidemiology, especially if accompanied by a behavioural response. Likely declines in bacterial causes of genital ulcers should be considered in defining policies on syndromic STD management in severe HIV epidemics. PMID- 12083449 TI - The value of mass screening for chlamydia control in high prevalence communities. AB - The social context and epidemiology of STIs in remote communities in Northern Canada was examined. These communities have a persistently high reported incidence of gonorrhoea and chlamydial infection. They remain in the hyperendemic phase of the N. gonorrhoeae and C. trachomatis epidemics. They are ethnically and culturally homogeneous and contain no readily identifiable core groups, making it impossible to distinguish between spread and maintenance networks. Mass screening of the adult population can reduce the reservoir of C. trachomatis infection under these circumstances. It is particularly important to target men in settings such as this where women are routinely screened in antenatal and family planning clinics. PMID- 12083450 TI - Role of core and bridging groups in the transmission dynamics of HIV and STIs in Cotonou, Benin, West Africa. AB - The potential for exposure of low and high risk women to HIV and sexually transmitted infections (STI) through unprotected sex with male clients of female sex workers in Cotonou could account for most if not all of the estimated yearly numbers of HIV infections in Cotonou women (approximately 1000). As ongoing transmission of HIV, and also of the most predominant STIs such as gonorrhoea and HSV-2, appears to be largely fuelled by transmission within core and bridging groups in Cotonou, interventions targeted at both female sex workers and their male clients remain of the utmost importance and could have a significant effect on the evolution of HIV/STI epidemics in Benin. PMID- 12083451 TI - The impact of HIV epidemic phases on the effectiveness of core group interventions: insights from mathematical models. AB - Mathematical models have highlighted the disproportionate contribution of core group transmitters to the spread of sexually transmitted diseases. Because the effectiveness of interventions varies with time, it has been suggested that epidemic phases should be considered in the design of prevention strategies. This study aimed to examine the impact of HIV epidemic phases on the effectiveness of HIV interventions based on gonorrhoea screening and condom use, targeted to core groups. The results are based on a mathematical model of gonorrhoea and HIV transmission in a relatively slow spreading HIV epidemic using Cotonou (Benin) as an example. For epidemics with a low reproductive potential modest core group interventions can significantly reduce HIV incidence and prevalence. As the epidemic matures, effective interventions should also incorporate core and non core populations. For epidemics with a high reproductive potential, core group interventions are necessary but not sufficient to have a rapid and large scale impact. A more general population approach is also needed early in the epidemic. Epidemic phases are also important in the evaluation of prevention strategies. PMID- 12083452 TI - STD/HIV intervention and research programme Mwanza Region, NW Tanzania. AB - The social determinants and epidemiology of sexually transmitted disease (STD) were studied in rural communities in Mwanza Region, Tanzania, in the context of the phase specific model of STD transmission. The prevalence of HIV and syphilis was higher in communities close to main roads, and lower in communities living on islands in Lake Victoria, probably reflecting the proportion of high risk individuals in the population. The prevalence of Herpes simplex virus type 2 infection, gonorrhoea, chlamydial infection, and trichomoniasis was similar in all types of community, reflecting the fact that these infections remain in the hyperendemic phase. The transmission of STDs is fuelled by high population mobility and by the presence of high risk individuals in rural as well as roadside communities. PMID- 12083453 TI - Sociodemographic context of the AIDS epidemic in a rural area in Tanzania with a focus on people's mobility and marriage. AB - This analysis focuses on how sociocultural and economic characteristics of a poor semi-urban and rural population (Kisesa ward) in north west Tanzania may directly and indirectly affect the epidemiology of HIV and other sexually transmitted infections (STI). Poverty and sociocultural changes may contribute to the observed high levels of marital instability and high levels of short and long term migration in Kisesa, especially among younger adults. Marriage and migration patterns are important underlying factors affecting the spread of HIV. The most cost-effective intervention strategy may be to focus on the trading centre in which mobility is higher, bars were more common, and HIV prevalence and incidence were considerably higher than in the nearby rural villages. If resources suffice, additional work can be undertaken in the rural villages, although it is not clear to what extent the rural epidemic would be self sustaining if the interventions in the trading centre were effective. PMID- 12083454 TI - Flavonoids in cell function. PMID- 12083456 TI - Inhibition of colonic aberrant crypt formation by the dietary flavonoids (+) catechin and hesperidin. PMID- 12083455 TI - Polyphenol antioxidants in citrus juices: in vitro and in vivo studies relevant to heart disease. AB - It is well known that eating fruits and vegetables lowers the risk of chronic diseases such as heart disease and cancer. The question of what is/are the active ingredient(s) is still unresolved. The initial hypothesis was that the antioxidant vitamins were responsible. However, recently the polyphenols have been investigated since they have been found to have beneficial properties such as being strong antioxidants. We measured the polyphenol content of citrus juices by an oxidation-reduction colorimetric method (Folin) using catechin as the standard. The order was tangerine juice > grapefruit juice > orange juice. The antioxidant contribution of ascorbic acid was measured by the difference in Folin reactive content following removal by ascorbate oxidase. Ascorbate contributed 56 to 77% of the antioxidant content of orange juice, 46% of the single tangerine juice measured, and 66 to 77% of grapefruit juices. Polyphenol quality in the juices was analyzed by using the inhibition of lower density lipoprotein oxidation promoted by cupric ion, an in vitro model of heart disease. Quality decreased in the following order: orange juice > grapefruit juice > tangerinejuice. In orange juice polyphenols accounted for 84-85% of antioxidant quality. The pure polyphenol hesperidin, which is common in juices, ascorbic acid, and the citrus juices, were not able to bind with LDL+VLDL and protect it from oxidation. In a hamster model of atherosclerosis, the juices were able to significantly inhibit atherosclerosis and lowered cholesterol and triglycerides. Ascorbic acid alone in the dose provided by the juices was found to have the same effect on atherosclerosis. However, the polyphenols in the citrus PMID- 12083457 TI - The citrus methoxyflavone tangeretin affects human cell-cell interactions. AB - Two effects of the citrus methoxyflavone tangeretin on cell-cell interactions are biologically relevant. Firstly, tangeretin upregulates the function of the E cadherin/catenin complex in human MCF-7/6 breast carcinoma cells. This leads to firm cell-cell adhesion and inhibition of invasion in vitro. Secondly, tangeretin downregulates the interleukin-2 receptor on T-lymphocytes and natural killer cells. This leads to a decrease in the cytotoxic competence of these immunocytes against cancer cells. The second effect can become important when high doses of tangeretin are combined with adjuvant tamoxifen treatment for breast cancer. Experiments with nude mice bearing MCF-7/6 tumors showed that tangeretin given orally at high doses, abrogated the therapeutic suppression of tumor growth exerted by tamoxifen. No evidence for a tumor promoting effect of tangeretin by itself was found in these experiments. Tangeretin may be an interesting molecule for application in cases where immunosuppression could be clinically beneficial. PMID- 12083458 TI - Xanthine oxidase and xanthine dehydrogenase inhibition by the procyanidin-rich French maritime pine bark extract, pycnogenol: a protein binding effect. PMID- 12083459 TI - Human 17beta-hydroxysteroid dehydrogenase type 5 is inhibited by dietary flavonoids. AB - Phytoestrogens contained in a vegetarian diet are supposed to have beneficial effects on the development and progression of a variety of endocrine-related cancers. We have tested the effect of a variety of dietary phytoestrogens, especially flavonoids, on the activity of human 17beta-hydroxysteroid dehydrogenase type 5 (17beta-HSD 5), a key enzyme in the metabolism of estrogens and androgens. Our studies show that reductive and oxidative activity of the enzyme are inhibited by many compounds, especially zearalenone, coumestrol, quercetin and biochanin A. Among flavones, inhibitor potency is enhanced with increased degree of hydroxylation. The most effective inhibitors seem to bind to the hydrophilic cofactor binding pocket of the enzyme. PMID- 12083461 TI - Regulation of lipoprotein metabolism in HepG2 cells by citrus flavonoids. PMID- 12083460 TI - Interactions of flavones and other phytochemicals with adenosine receptors. AB - Dietary flavonoids have varied effects on animal cells, such as inhibition of platelet binding and aggregation, inhibition of inflammation, and anticancer properties, but the mechanisms of these effects remain largely unexplained. Adenosine receptors are involved in the homeostasis of the immune, cardiovascular, and central nervous systems, and adenosine agonists/antagonists exert many similar effects. The affinity of flavonoids and other phytochemicals to adenosine receptors suggests that a wide range of natural substances in the diet may potentially block the effects of endogenous adenosine. We used competitive radioligand binding assays to screen flavonoid libraries for affinity and a computational CoMFA analysis of flavonoids to compare steric and electrostatic requirements for ligand recognition at three subtypes of adenosine receptors. Flavone derivatives, such as galangin, were found to bind to three subtypes of adenosine receptors in the microM range. Pentamethylmorin (Ki 2.65 microM) was 14- to 17-fold selective for human A3 receptors than for A1 and A2A receptors. An isoflavone, genistein, was found to bind to A1 receptors. Aurones, such as hispidol (Ki 350 nM) are selective A1 receptor antagonists, and, like genistein, are present in soy. The flavones, chemically optimized for receptor binding, have led to the antagonist, MRS 1067 (3,6-dichloro-2'-(isopropoxy)4' methylflavone), which is 200-fold more selective for human A3 than A1 receptors. Adenosine receptor antagonism, therefore, may be important in the spectrum of biological activities reported for the flavonoids. PMID- 12083462 TI - Anti-inflammatory actions of a micronized, purified flavonoid fraction in ischemia/reperfusion. AB - It is now recognized that reperfusion after a prolonged period of reduced or absent blood flow, although necessary to salvage ischemic tissue, initiates a complex series of deleterious reactions which ultimately induce the same effects as ischemia per se, i.e., cell injury and necrosis. Work conducted over the past 15 years has uncovered the fact that post-ischemic leukocyte infiltration plays a major role in the reperfusion component of ischemia/reperfusion (I/R) injury. This discovery has led to a concerted research effort directed at identifying interventions that prevent post-ischemic leukocyte adhesion and emigration. Recent work indicates that flavonoids are particularly effective anti inflammatory agents in the setting of I/R. While the mechanisms underlying the powerful protective effects of these compounds is uncertain, a growing body of evidence indicates that flavonoids are potent anti-oxidants that also act to inhibit the activity of key regulatory enzymes involved in the activation of pro inflammatory signaling cascades. In addition, it appears that these compounds prevent the expression of specific adhesion molecules involved in leukocyte recruitment, observations which provide the molecular basis for the anti-adhesive properties of these compounds. PMID- 12083463 TI - Flavonoids and gene expression in mammalian cells. AB - Flavonoids appear to regulate the expression of many genes. As expected, when multiple flavonoids were compared in one study, structure-specificity was always observed. Unfortunately, little information is available regarding the proportion of contribution of various structural elements. Also, we have very limited information on their molecular mechanisms of action. The affinity of flavonoids for ER could explain the stimulatory effect on genes with ERE but other modes of action apparently also exist and need to be further explored. Physiological relevance is always a concern when investigating the regulation of gene expression by environmental chemicals such as flavonoids. One factor of concern is the in vivo concentration of flavonoids. Besides intestinal cells, liver cells and skin cells, other tissues obtain flavonoids through blood circulation. Thus, plasma concentrations of flavonoids are normally discussed. Steady state plasma concentrations of flavonoids are usually not much higher than 1 microM even in populations that consume large amounts of plant material. This concentration is relatively low compared to the concentrations of flavonoids that were commonly used in cell culture systems to demonstrate their effectiveness. Nevertheless, we have evidence that some flavonoids may accumulate in the cell. The effect of quercetin on metallothionein expression in Caco-2 cells persisted for at least 24 hours after its removal from the culture medium (Kuo et al., 1998). Also, long term treatment of cultured cells with quercetin at low concentrations led to a similar effect on metallothionein expression as one high concentration treatment (Kuo et al., 2001). If intracellular accumulation of certain flavonoids is a shared characteristic for various cell types, it implies that routine ingestion of flavonoids could lead to biological effects at the concentration lower than predicted from a single treatment. Experiments to address possible cell/tissue accumulation of flavonoids are greatly needed. PMID- 12083464 TI - Signalling in arbuscular mycorrhiza: facts and hypotheses. PMID- 12083465 TI - The use of a photoactivatable kaempferol analogue to probe the role of flavonol 3 O-galactosyltransferase in pollen germination. AB - Flavonol induced pollen germination in petunia is rapid, specific, and achieved at low concentrations of kaempferol or quercetin. To determine the macromolecules that interact with the flavonol signal we have synthesized affinity-tagged kaempferol analogues. The first generation molecules are based on a benzophenone photophore. We find that 2-(3-benzoylphenyl)-3,5,7-trihydroxychromen-4-one (BPKae) antagonizes flavonol-induced pollen germination in a concentration dependent manner. Further, BPKae acts as an irreversible inhibitor of flavonol 3 O-galactosyltransferase (F3GalTase), the gametophyte-specific enzyme that controls the accumulation of glycosylated flavonols in pollen. The effects of BPKae are mediated by UV-A light treatment. The binding characteristics of BPKae to F3GalTase suggest that it can be used to identify the residues required for flavonol-binding and catalysis. PMID- 12083466 TI - Flavonoids: signal molecules in plant development. PMID- 12083467 TI - Modern analytical techniques for flavonoid determination. AB - We stand at the dawn of the next great period of natural products research. We now have tools to begin unraveling the chemical and physiological mechanisms by which these phytochemicals affect the function of living cells, both in plants and animals. The advances in analytical research will allow us to better use these compounds in the control of diseases in both plants and animals. This chapter used the analysis of soy isoflavones to illustrate the approaches to natural product research, but these principles can be applied to nearly any of the phenolic compounds found in plants. PMID- 12083468 TI - HPLC-mass spectrometry of isoflavonoids in soy and the American groundnut, Apios americana. PMID- 12083469 TI - History as a tool in identifying "new" old drugs. AB - To trace the history of a natural product and its use, it is necessary to identify to correct plant among around a half-million species. One must also know how and when harvest the plant and the morphology of location and extraction. Within the same species plant chemistry varies, depending upon climatic and soil conditions, stage of maturity and even diurnal factors. To all of these variations must be added the diagnostic ability of physicians and native healers (to distinguish between Hippocratically-trained Western physicians and whose knowledge is less formally taught). Seldom was a disease identified as we Know it today, but the constellations of symptoms described, when studied carefully within the framework historical setting of the culture, can be related to modern medicine. It is essential to study the historical contemporary usage data in the language in which those accounts were writTen. Translators are often philologists who are not sensitive to medical nuances. Modern readers of translated historical documents often are unaware of the precision the authors delivered in describing medical afflictions and their treatments. Natural product drugs are truly products of human knowledge. Because so many modern pharmaceuticals are manufactured synthetically we forget that once either the compound or its affinity had a home in a natural product. Over 2,500 years ago man first used a drug obtained from white willow bark, which was aspirin or acetylsalicylic acid. Today's scientists continue to be bewildered by just what aspirin's mechanisms of action are, discovering new modes of action, and how they relate to medical diagnostics. Whatever the science of aspirin, an intelligent person today takes it just as our ancestors did fo millennia. Throughout time, explanations continue to vary just as purpose of administration do as well. Nevertheless, aspirin is perceived as being beneficial. Historical in-use data can also be a factor in judging a drug's safety, since the records of its use provide observations made by intelligent persons over generations of employment Many historical "drugs" have crossed the line from drug to food. A number of them are now common items on our tables: coffee, tea, sugar, lemon, chocolate, pepper, to name a few The example of coffee affords useful insight. It was first employed as a drug (like tobacco), its botany and chemistry are well known, it has been in widespread use for centuries with diverse ethnic populations in a variety of preparations and amounts consumed. Still we are unsure about coffee's effect on health, the latest assertion being that the caffeine it contains may delay the onset of Alzheimer's. In contrast, the mercury drugs were in widespread use for a long period of time by many populations and that fact indicates that the toxic tolerance in humans is probably higher than as currently proscribed. The past contains important data for the scientific investigator. Like any field of research, historical investigation requires specialized knowledge, but much of that knowledge is readily accessible and employable. Rediscovery through examination of historical contemporaneous use data can be efficient and relatively easy compared to the travails of original research in pursuit of a discovery only to learn later that our ancestors had already made that discovery through trial and error in human usage. If we had started a search from the clues provided by history, presumably our discoveries would have been earlier, and we would have benefitted. As it is. we learn history but not science or else we learn science, not history. Both taken together the learning can be enhanced. PMID- 12083470 TI - Host recognition by pathogenic fungi through plant flavonoids. AB - A common characteristic among fungal pathogens of plants is that each specializes on a narrow range of specific plants as hosts. One adaptation to a specific host plant is the recognition of the host's chemicals which can be used to trigger genes or developmental pathways needed for pathogenesis. The production of characteristic flavonoids by plants, particularly those exuded from roots by legumes, appear to be used as signals for various microbes, including symbionts as well as pathogens. Nectria haematococca MPVI (anamorph: Fusarium solani) is a soil-borne pathogen of garden pea (Pisum sativum) which serves as a useful model in studying host flavonoid recognition. This fungus displays flavonoid induction of specific pathogenicity genes as well as stimulation of development needed for pathogenesis. Here, we summarize the study of flavonoid-inducible signal pathways which regulate these trait, through identification of transcription factors and regulatory components which control these responses. The characterization of the components a pathogen uses to specifically recognize its host provides insights into the host adaptation process at the molecular level. PMID- 12083471 TI - Potential health benefits from the flavonoids in grape products on vascular disease. AB - In the dog, monkey, a nd human we have shown that 5 ml/kg of red wine or 5-10 ml/kg of purple grape juice but not orange or grapefruit juice inhibits platelet activity, and protects against epinephrine activation of platelets. Red wine and purple grape juice enhances platelet and endothelial production of nitric oxide (Fitzpatrick et al., 1993, Parker et al., 2000). This is thought to be one of the mechanisms whereby purple grape juice significantly improved endothelial function in 15 patients with coronary artery disease. The consumption of purple grape juice by the patients also offered increased protection against LDL cholesterol oxidation, even though all the patients were also taking another antioxidant vitamin E, 400 IU/day. The number of people and animals in these studies was small; however, each one acted as their own control as measurements were made in each before, and then after consumption of red wine or purple grape juice. Thus these studies are thought to be significant. We feel that the results of these studies are encouraging and justify further research on larger numbers of subjects. This suggests that the flavonoids in purple grape juice and red wine may inhibit the initiation of atherosclerosis by one or more of the mechanisms described above. It will take years to fully characterize the potential benefits of daily consumption of red wine or purple grape juice for maintaining a healthy heart. Based on the existing evidence of antiplatelet and antioxidant benefits and improved endothelial function from red wine and purple grape juice, it seems reasonable to suggest that moderate amounts of red wine or purple grape juice be included among the 5-7 daily servings of fruits and vegetables per day as recommended by the American Heart Association to help reduce the risk of developing cardiovascular disease. PMID- 12083472 TI - What are your patients reading? PMID- 12083473 TI - Developing cataract surgery priority criteria: results from the Western Canada Waiting List Project. AB - BACKGROUND: The Western Canada Waiting List Project (WCWL) is a federally funded partnership of 19 organizations, including medical associations, health authorities, ministries of health and research organizations, that was created to develop tools to assist in the management of waiting lists. The WCWL cataract surgery panel, one of five panels constituted under this project, developed and tested a set of standardized clinical criteria for prioritizing among patients awaiting cataract surgery. METHODS: The cataract surgery panel was composed of seven academic and community ophthalmologists, two family physicians, an optometrist, a health care services researcher and a health information specialist. The panel met three times between October 1999 and June 2000. The priority criteria were applied to 563 patients in the four western provinces. Regression analysis was used to determine the set of criteria weights that collectively best predicted clinicians' overall ratings of urgency. Interrater and test-retest reliability were assessed, based on clinicians' ratings of videotaped interviews with six hypothetical patients. RESULTS: The resulting criteria accounted for about one-third of the observed variance in clinicians' ratings of overall clinical urgency (R2 = 31.2%). The panel modified the original criteria and weights based on empirical results and clinical judgement. Reliability of the revised criteria items was mixed, with the lowest reliability observed for items dealing with glare and social role. INTERPRETATION: Participating clinicians considered the criteria easy to use and reasonably reflective of expert surgical judgement regarding clinical urgency. Further development and testing of the tool appear warranted. PMID- 12083475 TI - The effect of a hockey visor and sports goggles on visual function. AB - BACKGROUND: Despite the potential for eye injury, most hockey players in the National Hockey League do not use a protective face shield. We examined the effect of a hockey visor and sports goggle on visual function to see whether visual impediment is a substantial factor in the resistance to use of protective eyewear. METHODS: We measured Snellen visual acuity, Ishihara colour vision and contrast sensitivity and performed Humphrey central 30 degrees and peripheral 30 degrees to 60 degrees threshold perimetry in eight volunteers (10 eyes) with and without a sports goggle and a hockey visor (half face shield). Subjects with normal acuity and normal confrontation fields who did not require presbyopic correction at near were enrolled. Five subjects (10 eyes) had further testing with threshold perimetry of the temporal crescent. The order in which the tests were performed (no mask first, visor first or goggle first) was alternated between subjects. RESULTS: There was no difference in Snellen acuity or Ishihara colour vision, and no statistically significant difference in contrast sensitivity or foveal threshold with or without protective eyewear. The visor caused a statistically significant decrease in the central 30 degrees mean deviation index compared to no mask (p = 0.001), but the goggle did not. Each of the peripheral quadrant threshold totals was significantly decreased with the visor compared with no mask (p < 0.05). The 30 degrees to 60 degrees temporal field was better preserved with the goggle than with the visor, but the nasal peripheral field was worse with the goggle than with the visor. Both the visor and the goggle caused a statistically significant decrease of more than 4 decibels in the temporal crescent threshold (p = 0.000). INTERPRETATION: Protective eyewear does not adversely affect Snellen acuity, contrast sensitivity, Ishihara colour vision or foveal threshold. The central 30 degrees field of vision is not affected by the sports goggle and is minimally depressed by the hockey visor. The goggle better preserves the temporal midperipheral field, but, unlike the visor, the goggle caused a monocular nasal lens rim scotoma. Both devices caused a depression of more than 4 dB of the far temporal field beyond 60 degrees (temporal crescent). PMID- 12083474 TI - The Misericordia Health Centre cataract comfort study. AB - BACKGROUND: All surgery provokes various degrees of anxiety for patients. The environment leading up to surgery can affect anxiety levels. We performed a prospective randomized study to compare environmental factors around the time of cataract surgery in order to identify interventions that would minimize stress for patients. METHODS: Patients scheduled to undergo cataract surgery at a university-affiliated hospital in Winnipeg were randomly assigned to 1) receive orally administered lorazepam or a placebo before surgery; 2) listen to relaxing music through headphones or routine background noise before surgery; 3) walk (or go by wheelchair if unable to walk) to the operating room or go by stretcher; and 4) listen to relaxing music through headphones or routine background noise during surgery. Randomization for part 1 was double blind; for parts 2 and 3 the surgeon and anesthetist were blinded, but the patient was not. Patients were asked to rate their anxiety, sedation, nausea and pain on arrival at the preoperative area, about 30 minutes after arrival, on arrival in the operating room and on arrival in the postoperative area, on a visual analogue scale graded from 0 ("None" [or "Wide awake" in the case of sedation]) to 10 ("Worst possible" [or "Asleep" in the case of sedation]). Patient satisfaction and willingness to repeat the exact same form of treatment were also rated. RESULTS: Of the 19 surgeons in the department 18 agreed to participate; I withdrew during the study. Data were collected for 144 patients aged 26 to 93 years. Anxiety was highest on arrival at the institution and decreased progressively thereafter. Oral sedation and listening to music before surgery were associated with decreased anxiety and increased levels of sedation (p = 0.002). Walking to the operating room provided no benefit over going by stretcher. Listening to music through headphones during surgery was not accepted by many patients and, when used, negatively affected the surgeon's assessment of the patient's ability to cooperate. Surgeons reported movement more often among patients who received oral sedation than among those who did not (chi2 = 0.01). Levels of pain and nausea were extremely low in all patients, and satisfaction was very high. Patients who received regional local anesthesia had less pain and higher satisfaction than those who received topical anesthesia. Willingness to repeat the same treatment was extremely high. INTERPRETATION: For patients undergoing cataract surgery, efforts should be directed toward reducing anxiety on arrival at the institution, when it is highest, and not just during surgery. Oral sedation and listening to music before surgery appear to be beneficial. Listening to music through headphones during surgery was not found to be advantageous. PMID- 12083476 TI - Ocular adnexal injuries from industrial blunt hook trauma. PMID- 12083477 TI - The role of conjunctival biopsy in the diagnosis of Wegener's granulomatosis: a case report. PMID- 12083478 TI - Retinal venous beading associated with recurrent branch vein occlusion. PMID- 12083479 TI - Polymorphonuclear leucocytes have two opposing roles in fibrinolysis. AB - Polymorphonuclear leucocytes (PMN) are important in the resolution of human thrombi, with u-PA as a key player. We have shown that the u-PA activity of PMN depends on the presence of plasma; the study presented here provides an explanation for that requirement. Here we show that PMN degraded scu-PA and also tcu-PA, t-PA and plasmin, resulting in loss of fibrinolytic activity. Plasma protected against this degradation; alpha1-antitrypsin was identified as a protective factor. Purified human neutrophil elastase mirrored the effects of PMN, again neutralized by plasma inhibitors. These findings illustrate the dual role of PMN in the breakdown of thrombi, in that they contribute both u-PA, which lyses fibrin, and other proteases, including elastase, which can cleave fibrin and plasminogen activators/plasmin. Similarly, plasma can potentiate fibrinolysis by neutralization of PMN elastase, in addition to direct inhibition of fibrinolytic proteases. Our previous studies show that PMN in thrombi are mostly pro-fibrinolytic; the anti-fibrinolytic role defined here may be important in other pathologies where fibrin persists. PMID- 12083480 TI - In vivo plasminogen deficiency reduces fat accumulation. AB - Obesity and non-insulin dependent diabetes are associated with a decrease in fibrinolysis, which is mediated by the plasminogen system. The purpose of the current study was to investigate the role of the plasminogen system in the reduced body weight of the plasminogen deficient (Plg-/-) mice. In this study we have found that the reduced body weight in Plg-/- mice is due to a reduced rate of the adipose tissue (25% less) and whole body fat (30% less) accumulation during growth in Plg-/- compared to wild-type (WT) littermates. When the mice are fed a high fat-lipogenic diet, adipose tissue accumulation increases in the Plg-/ mice indicating that the capacity for lipid filling of cells was not blocked. In addition, glycerol phosphate dehydrogenase, a marker of late differentiation, was not different in the depots from WT and Plg-/- mice. The number of stromal cells (number x 10(5)/g adipose tissue), isolated from inguinal (Plg-/- 3.4 +/- 1.2. n = 6; WT 0.17 +/- 0.07, n = 7, p < 0.02) and gonadal (Plg-/- 11.0 +/- 0.4, n = 6; WT 3.1 +/- 0.7, n = 7, p < 0.05) fat depots. was markedly higher in the depots from the Plg-/- mice than WT mice. Differentiation of stromal cells in culture from the Plg-/- mice was reduced compared to cells from WT mice. These results suggest that differences in the stromal cell population are responsible for the reduced adipose tissue accumulation in the Plg-/- mice, and that the plasminogen system plays an important role in adipose tissue accumulation. PMID- 12083481 TI - An additional mechanism of action of abciximab: dispersal of newly formed platelet aggregates. AB - BACKGROUND: The ability of abciximab to prevent fibrinogen binding to activated platelets indicates it may also promote dissolution of platelet-rich thrombi. The present study examined the capacity of abciximab to reverse platelet aggregation in vitro. METHODS AND RESULTS: Experiments were performed on blood from healthy non-medicated donors. Platelet aggregate formation and disaggregation were monitored turbidimetrically. Platelet-bound fibrinogen was measured by flow cytometry. For disaggregation studies, platelets were first stimulated with either ADP or the 11-mer thrombin receptor activating peptide (TRAP), then varying amounts of abciximab were added at periodic intervals after agonist addition. Platelet disaggregation was detected by comparing the extent of light transmittance at 4 min after addition of either abciximab or saline to PRP. ATP release was simultaneously monitored by chemi-luminescence. When added 1 min after low concentrations of ADP, abciximab rapidly (< 1 min) dispersed platelet aggregates in a dose-dependent manner, with complete disaggregation observed with 6.25 microg/mL of the beta3 antagonist. In contrast, equivalent concentrations of abciximab did not induce appreciable disaggregation to platelets stimulated with TRAP (10 microM). Platelet counts of samples that had undergone complete disaggregation, as assessed by aggregometry, were equivalent to baseline, indicating dispersal of aggregates to single cells. Concentrations of abciximab that produced complete disaggregation induced partial displacement of platelet bound fibrinogen (52 +/- 8% inhibition of fibrinogen binding at 12.5 microg/ml abciximab). The disaggregation effectiveness of abciximab decreased as the time between ADP and subsequent abciximab addition widened, and as the amount of both dense granule release and agonist stimulation increased. However, pre-treatment of platelets with acetylsalicylic acid (ASA) did not potentiate platelet disaggregation induced by abciximab. CONCLUSIONS: These data indicate that abciximab facilitates the dispersal of newly formed platelet aggregates in vitro, by partially displacing fibrinogen from activated GPIIb/IIIa receptors. In vivo, abciximab may destabilize coronary thrombi by preventing aggregate formation and dispersing mural thrombi. PMID- 12083482 TI - Selective inhibition of protease-activated receptor 4-dependent platelet activation by YD-3. AB - In the present study, the antiplatelet effect and its mechanism of a new synthetic compound YD-3 [1-benzyl-3-(ethoxycarbonylphenyl)-indazole] were examined. YD-3 inhibited the aggregation of washed human platelets caused by protease-activated receptor (PAR) 4 agonist peptide GYPGKF (IC50 = 0.13 +/- 0.02 microM), but had no or little effect on that by thrombin, PAR1 agonist peptide SFLLRN, collagen or U46619. YD-3 produced a parallel, rightward shift of the concentration-response curve for GYPGKF without decreasing of the maximum platelet aggregation, indicating a competitive antagonism. In contrast to human platelets, both thrombin- and GYPGKF-induced mouse platelet shape change and aggregation were completely inhibited by YD-3. YD-3 also selectively prevented GYPGKF-induced intracellular Ca2+ mobilization in human platelets. Furthermore, in the PAR1-desensitized human platelets, thrombin induced a relatively slow rise and decay of calcium mobilization that was significantly inhibited by YD-3. In addition, the synergistic effect of SFLLRN and GYPGKF on platelet activation was prevented by YD-3. YD-3 also inhibits both fMLP-stimulated neutrophil- and purified cathepsin G-induced platelet aggregation, which has been demonstrated to be PAR4-dependent. Taken together, our results suggest that YD-3 selectively inhibits PAR4-dependent platelet activation through blockade of PAR4. To the best of our knowledge, it is the first non-peptide PAR4 antagonist. PMID- 12083483 TI - Glanzmann's thrombasthenia: identification of 19 new mutations in 30 patients. AB - Glanzmann's thrombasthenia (GT) is a genetically heterogeneous autosomal recessive syndrome associated with a bleeding tendency. To elucidate molecular basis of GT we have screened for mutations 30 GT patients. On the whole, 21 different candidate causal mutations, 17 in the alphaIIb and 4 in the beta3 gene have been found. Only two (alphaIIb Pro145Ala and IVS3(-3)-418del) have been previously reported. Nine mutations (42.9%) were likely to produce truncated proteins, whereas the remaining 12 were missense mutations that affected highly conserved residues in alphaIIb and beta3 genes. Six mutations were found in different patients suggesting a possible founder effect. The wide spectrum of expressivity, ranging from mild to severe also among patients carrying the same mutations, provided evidence for a role of different loci or circumstantial factors. In conclusion, we have identified a spectrum of unreported mutations that may be of value to unravel the role of specific regions of alphaIIb and beta3 genes. PMID- 12083484 TI - FAK-mediated inhibition of vascular smooth muscle cell migration by the tetraspanin CD9. AB - Migration of vascular smooth muscle cells (SMC) towards the intima is a key event in vascular proliferative diseases. We investigated a potential role for the tetraspanin CD9 in this process in a wound migration assay. Aortic SMC from CD9 knock-out mice had higher migration rates and the presumably stimulatory anti-CD9 antibody ALMA-1 inhibited migration of human SMC. The signaling pathways responsible for this inhibitory effect were investigated. In migrating CD9-/- SMC, stress fiber formation was decreased and focal adhesions were smaller and more diffusely distributed, consistent with an inhibition of integrin clustering. In migrating mouse SMC expressing CD9, focal adhesion kinase (FAK) tyrosine phosphorylation was doubled. No differences in intracellular calcium signaling were observed between CD9+/+ and CD9-/- SMC during migration. We suggest that CD9 in hibits SMC migration by a stimulation of both stress fiber formation and integrin clustering, leading to a stimulation of FAK phosphorylation. PMID- 12083485 TI - The expression of tissue factor and tissue factor pathway inhibitor in aortic smooth muscle cells is up-regulated in synthetic compared to contractile phenotype. AB - Tissue factor (TF) and its specific inhibitor TF pathway inhibitor (TFPI) are produced by vascular smooth muscle cells (SMCs) in vitro and are increased in vivo in atherosclerotic compared to normal vessels. Besides local regulation of the hemostatic balance, this may be related to non-hemostatic TF/protease dependent functions such as SMC proliferation, adhesion and migration. The aim of the study was to compare the expression of both proteins between the contractile (normal adult) and synthetic (neo-intimal) SMC phenotypes. Primary cultures of SMCs isolated from rat thoracic aorta before and 10 days after balloon injury displayed stable characteristics of the contractile and synthetic phenotype, respectively. Synthetic SMCs expressed more TF mRNA than contractile SMCs, but released excess TF in the conditioned medium, so that the cell-associated TF activity measured by a factor Xa generating assay remained similar in the two subtypes. Accordingly, cell surface thrombogenicity measured under blood flow conditions was also similar. The production and release of functional TFPI was enhanced by a factor 3 to 6 (p < 0.01) in synthetic SMCs. A difference in the quantitative expression of TF and TFPI is a new distinctive feature of SMC phenotypes. Matrix-associated TFPI derived from synthetic SMCs may serve as an anchorage for their migration and regulate protease-activated processes during neo-intima formation. PMID- 12083486 TI - Factor VIIa induced release of von Willebrand factor from human umbilical vein endothelial cells by a tyrosine kinase dependent pathway. AB - The interaction of FVIIa with surface-bound tissue factor (TF) induces various cellular changes including cytosolic Ca2+ signals. The release of von Willebrand factor (VWF) from endothelial cell stores may be triggered by an elevation in cytosolic free Ca2+, therefore we investigated the effect of rFVIIa on the release of VWF from human umbilical vein endothelial cells (HUVEC). We show here that rFVIIa induces the release of VWF from HUVEC with or without prestimulation with lipopolysaccharide (LPS). The effect of rFVIIa was dose dependent. However, the release of VWF by HUVEC in response to rFVIIa was significantly greater with LPS prestimulation (3.18 times control) than without LPS prestimulation (1.45 times control) (p < 0.001). Cytosolic Ca2+ signals were detectable only after LPS prestimulation of HUVEC and these were small compared to those elicited by thrombin. No effect on rFVIIa induced release of VWF was seen in the presence of hirudin, site inactivated rFVIIa or the protein kinase C (PKC) inhibitor staurosporine. However, a tyrosine kinase inhibitor genistein, inhibited the rFVIIa induced release of VWF. These data show that release of VWF can occur without involvement of the cytosolic Ca2+/ PKC pathway. FVIIa induced VWF release from endothelial cells may have in vivo significance at sites of TF expression. PMID- 12083487 TI - Tissue factor activity is upregulated in human endothelial cells exposed to oscillatory shear stress. AB - Hemodynamic forces play a critical role in the pathogenesis of atherosclerosis as evidenced by the focal nature of the disease. Oscillatory shear stress characterizes the hemodynamic environment of plaque-prone areas as opposed to unidirectional shear stress typical of plaque-free areas. These particular flow conditions modulate atherosclerosis-related genes. Tissue factor (TF) initiates blood coagulation, contributes to vascular remodeling, and is therefore a potential contributor in the development/progression of atherosclerosis. We investigated the effect of oscillatory and unidirectional flows on TF using an in vitro perfusion system. Human endothelial cells exposed for 24 h to oscillatory shear stress, significantly increased TF mRNA, and TF protein expression (1.5- and 1.75-fold, respectively, p < 0.01), and surface TF activity (twofolds increase). Expression of TF inhibitor (TFPI), mRNA and protein, remained unchanged as compared to static conditions. Conversely, cells exposed to unidirectional shear, showed a decrease in TF activity with a significant increase in TFPI mRNA and protein expression (1.5- and 1.8-fold, respectively, p < 0.01). These results show for the first time that pulsatile oscillatory shear stress induces a pro-coagulant phenotype of endothelial cells which may favor formation/progression of atherothrombotic lesions. PMID- 12083488 TI - Function of tissue-type plasminogen activator releaser on vascular endothelial cells and thrombolysis in vivo. AB - The effect of monosodium[2-(6-hydroxynaphthalen-2-yl)-6-methylpyrimidin-4 yloxy]acetate dihydrate (JTV-926) on fibrinolysis was investigated in vitro and in vivo. JTV-926 released tissue-type plasminogen activator (t-PA) from human vascular endothelial cells in a dose-dependent manner. The thrombolytic effect of JTV-926 was studied using three animal thrombosis models; a photo-irradiation induced mouse carotid artery thrombosis model, a photo-irradiation-induced rat femoral artery thrombosis model and a thrombin-induced rat venous thrombosis model. In the mouse thrombosis model, t-PA deficient mice (t-PA-/- mice) and their wild-type (t-PA+/+) were used. JTV-926 was injected as a bolus 30 min after the interruption of blood flow by an occlusion thrombi. Blood flow was continuously monitored for 180 min after intravenous administration of JTV-926 (1 mg/kg). Although the recanalization rate of the occluded artery was 37.5% in t-PA +/+ mice with the vehicle control, it increased to 75% in t-PA+/+ mice after JTV 926 administration. However, when JTV-926 was administrated in t-PA-/- mice, vascular recanalization was not observed in any arteries. In the photo irradiation-induced rat femoral artery thrombosis model, intra-duodenal administration of JTV-926 induced thrombolysis. Moreover, in the thrombin-induced rat venous thrombosis model, the dose-dependent thrombolysis was also observed by oral administration of JTV-926. It was suggested that JTV-926 revealed a sufficient thrombolytic effect through the absorption from the intestine. Thus, a newly synthesized compound, JTV-926 induced t-PA release from vascular endothelial cells and effective thrombolysis in vivo. PMID- 12083489 TI - Effect of coenzyme Q10 and Ginkgo biloba on warfarin dosage in stable, long-term warfarin treated outpatients. A randomised, double blind, placebo-crossover trial. PMID- 12083490 TI - Variation in relative risk of venous thromboembolism in different cancers. PMID- 12083491 TI - Moderate hyperhomocysteinemia and central retinal vein occlusion. PMID- 12083492 TI - Factor V Leiden and prothrombin gene mutations in femoral head osteonecrosis. PMID- 12083493 TI - An extremely low prevalence of factor V Leiden, FIIG20210A and FXIIIV34L in Taiwan Chinese population. PMID- 12083494 TI - Changes in von Willebrand factor and factor VIII levels during the menstrual cycle. PMID- 12083495 TI - Correlation of activated clotting time (ACT) with rFVLLa infusion and clinical efficacy in a patient with Glanzmann thrombasthenia and platelet alloantibodies. PMID- 12083496 TI - Point mutations in the endothelial protein C receptor (EPCR) promoter. PMID- 12083497 TI - Blood clotting activation, angiogenesis and tumor metastasis: any role for TFPI? PMID- 12083498 TI - Tissue factor pathway inhibitor reduces experimental lung metastasis of B16 melanoma. AB - The importance of tissue factor (TF) in tumor biology has been highlighted by studies suggesting its involvement in cell signaling, metastasis and angiogenesis. Since many animal studies have shown that anticoagulant therapy can reduce experimental metastasis, we studied whether the natural inhibitor of TF mediated blood coagulation, Tissue Factor Pathway Inhibitor (TFPI), might be similarly effective. Using a murine experimental model, we found that intravenous injection of recombinant murine TFPI immediately before introduction of tumor cells reduced metastasis by 83% (P < 0.001). B16 murine melanoma cells stably transfected with a TFPI expression vector exhibited reduced lung seeding following intravenous injection by 81% (P < 0.001) compared with controls. No difference in primary tumor growth was observed between TFPI+ and control cells. Mice receiving intravenous somatic gene transfer of sense TFPI expression vector developed 78% fewer lung nodules than controls (P < 0.05). We conclude that TFPI has significant anti-metastatic activity in this experimental model. PMID- 12083499 TI - The vitamin K-dependent carboxylase. PMID- 12083500 TI - Randomised, double blind, multicentre, placebo controlled study of sulodexide in the treatment of venous leg ulcers. AB - Sulodexide, a highly purified glycosaminoglycan, was investigated for treatment of venous leg ulcers. Patients (n = 235) undergoing local treatment including wound care and compression bandaging, were randomised to receive either sulodexide or matching placebo for three months. Primary study endpoint was complete ulcer healing after 2 months; secondary endpoints were ulcer healing at 3 months and the time-course changes of ulcer areas. The proportion of patients with complete ulcer healing was higher with sulodexide at 2 months (p = 0.018) and 3 months. The "number needed to treat" to obtain one additional patient healed with sulodexide was 7 at 2 months and 5 at 3 months. The changes in ulcer surface area with time were significant for sulodexide only (p = 0.004). Fibrinogen significantly decreased in sulodexide patients (p = 0.006). In conclusion, sulodexide associated with local treatment proved to be effective and well tolerated in the management of venous leg ulcers. PMID- 12083501 TI - Tissue factor expression on monocyte subpopulations during normal pregnancy. AB - Pregnancy is often referred to as a hypercoagulable state due to changes in the haemostatic system. Tissue factor (TF) is the initiator of blood clotting in vivo. The effect of pregnancy on monocyte TF expression was determined in a longitudinal case control study (89 pregnant, 39 non-pregnant). Using whole blood flow cytometry and CD14 as a monocyte marker, TF expression was measured on all CD14 positive, CD14Bright and CD14Dim cells. TF expression was significantly lower in pregnant women than in non-pregnant control subjects, on all CD14 positive cells at 20 and 35 weeks, on CD14Bright cells at 12 and 35 weeks and on CD14Dim cells at 20 weeks. Additionally, we report that a higher percentage of CD14Dim than CD14Bright cells express TF. These results suggest that, in order to maintain homeostasis in haemostasis in an otherwise hypercoagulable state, monocyte TF expression is reduced during normal pregnancy. PMID- 12083502 TI - Fibrinogen predicts restenosis after endovascular treatment of the iliac arteries. AB - OBJECTIVE: Fibrinogen is an acute phase protein as well as a component of the coagulation cascade. Vascular inflammation and disturbed coagulation are suggested to cause restenosis after percutaneous transluminal angioplasty (PTA). We investigated the prognostic impact of fibrinogen on restenosis after endovascular treatment of iliac artery occlusive disease. METHODS: In a prospective cohort study 137 consecutive patients after iliac artery PTA (n = 74) and PTA plus selective stent implantation (n = 63) were included, 109 patients after lower limb angiography served as a control group. Patients were followed for 6 months with oscillography, ankle brachial index and duplex sonography for occurrence of restenosis. Fibrinogen and serum amyloid A (SAA), as a control parameter of inflammation, were obtained at baseline, 8, 24 and 48 h postintervention. RESULTS: PTA (adjusted OR 3.1, p = 0.05) and stenting (adjusted OR 13.3, p = 0.001) were independently associated with a higher postintervention increase of fibrinogen compared to angiography. Restenosis was found in 29 patients (21%). Patients with pre-intervention fibrinogen values in the third quartile (411-463 mg/dl) had a 6.2-fold increased adjusted risk for restenosis (p = 0.03), patients in the fourth quartile (> 463 mg/dl) had a 8.9-fold increased adjusted risk (p = 0.007). Pre-intervention SAA values were also significantly associated with restenosis (p < 0.0001). Postintervention fibrinogen and SAA levels showed no association with outcome. CONCLUSION: Balloon angioplasty and stenting of the iliac arteries cause an elevation of postintervention fibrinogen levels independently of angiographic factors. A higher pre-procedure fibrinogen level, presumably a marker of inflammatory activity, indicates a higher risk for restenosis. PMID- 12083503 TI - Increased angiostatin levels in bronchoalveolar lavage fluids from ARDS patients and from human volunteers after lung instillation of endotoxin. AB - Acute respiratory distress syndrome (ARDS) is characterized by a disruption of the alveolar-capillary barrier, due to both an epithelial and an endothelial dysfunction. Whereas epithelial apoptosis seems to be mainly mediated by Fas ligand, the mediators of endothelial damage remain to be identified. Angiostatin, a powerful inhibitor of angiogenesis in vivo, also specifically induces apoptosis in endothelial cells. The concentration of various enzymes that cleave angiostatin from plasminogen was reported to be significantly increased in bronchalveolar lavage (BAL) fluids from patients with ARDS. Therefore, in this study, we investigated whether angiostatin was generated during the pulmonary inflammatory response of both healthy subjects challenged with endobronchial endotoxin and in patients with ARDS. We found significantly elevated angiostatin levels in BAL fluids from patients at risk for and with early ARDS (up to 0.022% and 0.018% of total protein, respectively), as well as in BAL fluids from volunteers treated with endotoxin (up to 1.17% of total protein), as compared to BAL fluids from control patients (< 0.005% of total protein). These data suggest that angiostatin may contribute to the endothelial damage observed in ARDS, probably via an increased permeability of the alveolar capillary barrier, allowing for an intra-alveolar processing of its precursor plasminogen. PMID- 12083504 TI - Factor V Leiden and prothrombin gene G20210A mutation in children with venous thromboembolism. AB - To determine whether factor V Leiden (FVL) and/or prothrombin gene G20210A mutation (PT20210A) are risk factors for venous thromboembolism (VTE) in Argentinean children. One hundred and thirty consecutive children with VTE were prospectively assisted at a single centre. Blood samples were available from 110 of them for detailed haematological analysis. The prevalence of both mutations was compared with a control group. The odds ratio for VTE was significantly increased in patients with FVL (OR 3.64; 95% CI: 1.14-11.6, p < 0.029) whereas odds ratio for VTE was not significantly increased in patients with PT20210A (OR 1.06; 95% CI: 0.24-4.73, p = 0.938). Combined disorders were found in 5 of the 10 children with the aforementioned mutations. In 21 children (19%) without these mutations other inherited and acquired disorders were detected. Our data show that FVL is a risk factor for VTE whereas PT20210A does not seem to be a risk factor in our paediatric population. PMID- 12083505 TI - Anticoagulant and membrane-degrading effects of secretory (non-pancreatic) phospholipase A2 are inhibited in plasma. AB - Plasma concentrations of secretory (non-pancreatic) phospholipase A2 (sPLA2) may rise 1000-fold during inflammation, and this acute phase response has been related to anticoagulant effects. In the present study this hypothesis was further investigated. Prothrombinase activity was measured for model membranes mimicking the phospholipid composition of the outer membrane of resting and activated blood platelets. Using ellipsometry, membrane degradation by sPLA2 could be measured simultaneously with inhibition of thrombin production. The same technique was used to study clotting, by the sudden appearance of fibrin strands on the membrane. Results were compared with the effects of sPLA2 on the activation of washed platelets and platelets in plasma. In buffer solution, model membranes were degraded by (patho)physiological concentrations of sPLA2. Even when only partially degraded, membranes rapidly lost their prothrombinase activity, indicating preferential degradation of phosphatidylserine. Addition of diluted plasma interfered with membrane degradation, and also with inhibition of prothrombinase activity. In agreement with these observations, sPLA2 inhibited thrombin production and annexin V-binding of activated washed platelets, but had no effects on platelet activation or clotting in plasma. These findings indicate that the elevated plasma sPLA2 concentrations observed in inflammatory disease will not reduce hypercoagulability in such patients. PMID- 12083506 TI - Pharmacokinetics of new synthetic heparin mimetics. AB - Pharmacokinetics of oligosaccharides displaying various affinities for antithrombin (AT) allowed us to show that there was a close relationship between the plasma half-life of these antithrombotic oligosaccharides and their affinity for AT. Recently, we have described new heparin mimetics comprising an AT binding domain and a thrombin binding domain separated by a neutral methylated saccharide sequence. These compounds displayed strong anti-Xa and antii-IIA activities and, in contrast to heparin, escaped neutralisation by platelet factor 4. The aim of this work was to compare the pharmacokinetics of several of these heparin mimetics in rats. These compounds differed by their length, charge and affinity for AT (AT-binding domain). The results obtained indicate that the prolongation of the AT binding domain did not modify significantly their affinities for AT. However, an increase in the number of charges leads to a decrease in the half life. When a methylated spacer was added in order to minimise the non-specific interactions to other proteins, half-lives of the heparin mimetic were in the same range than that of the pentasaccharide used as an AT binding domain. In conclusion, this study shows the influence of the charge of the oligosaccharides on their pharmacokinetics and underlinesthe importance of minimising their non specific binding to plasma proteins in order to obtain compounds with predictive pharmacokinetics. PMID- 12083507 TI - Heterogeneous detection of A-antigen on von Willebrand factor derived from platelets, endothelial cells and plasma. AB - The exact function of the carbohydrate component of von Willebrand factor (VWF) is unknown. ABO blood group antigens are present as integral structures on the oligosaccharide side chains and it has long been recognised that ABO blood group is a determinant of VWF levels. The mechanism for this is not known. Using a monoclonal antibody against the A-antigen, we investigated the presence of this antigen on VWF from plasma, platelets, human umbilical vein endothelial cells (HUVEC) and saphenous vein endothelial cells. Initial studies on plasma VWF revealed that 23.5% of samples appeared to be negative for the A-antigen. This was shown to correlate with the A2 subtype of the A-antigen (p < 0.01). Analysis of intracellular VWF from saphenous vein endothelial cells revealed low levels of A-antigen to be present in comparison to the corresponding plasma VWF. In contrast, VWF from platelets and HUVEC gave no detectable A-antigen. However, within 1 h of administration of DDAVP to type 1 VWD patients, there was a > 2 fold increase in the A-antigen/VWF: Ag ratio for VWF in the plasma. In vitro experiments with serum N-acetlygalactosaminyltransferase failed to demonstrate any addition of A-antigen to platelet or HUVEC VWF. These data are consistent with heterogeneity in the content of A-antigen on VWF from different physiological compartments. Also, they are consistent with either a change in the A-antigen content of VWF after release from the intracellular compartment or a difference in the intracellular addition of A-antigen to VWF by endo thelium from different vascular beds. PMID- 12083508 TI - A performance evaluation of commercial fibrinogen reference preparations and assays for Clauss and PT-derived fibrinogen. AB - The wide availability of fibrinogen estimations based on the prothrombin time (PT Fg) has caused concern about the variability and clinical utility of fibrinogen assays. In a multi-centre study, we investigated fibrinogen assays using various reagents and analysers. Clauss assays generally gave good agreement, although one reagent gave 15-30% higher values in DIC and thrombolysis. Two commercial reference preparations had much lower potencies than the manufacturers declared, and plasma turbidity influenced parallelism in some Clauss assays. PT-Fg assays gave higher values than Clauss and showed calibrant dependent effects, the degree of disparity correlating with calibrant and test sample turbidity. Analyser and thromboplastin dependent differences were noted. The relationship between Clauss and PT-Fg assays was sigmoid, and the plateau of maximal PT-Fg differed by about 2 g/l between reagents. ELISA and immunonephelometric assays correlated well, but with a high degree of scatter. Antigen levels were higher than Clauss, but slightly lower than PT-Fg assays, which appeared to be influenced by degraded fibrinogen. Clauss assays are generally reproducible between centres, analysers and reagents, but PT-Fg assays are not reliable in clinical settings. PMID- 12083509 TI - Studying genomes through the aeons: protein families, pseudogenes and proteome evolution. AB - Protein families can be used to understand many aspects of genomes, both their "live" and their "dead" parts (i.e. genes and pseudogenes). Surveys of genomes have revealed that, in every organism, there are always a few large families and many small ones, with the overall distribution following a power-law. This commonality is equally true for both genes and pseudogenes, and exists despite the fact that the specific families that are enlarged differ greatly between organisms. Furthermore, because of family structure there is great redundancy in proteomes, a fact linked to the large number of dispensable genes for each organism and the small size of the minimal, indispensable sub-proteome. Pseudogenes in prokaryotes represent families that are in the process of being dispensed with. In particular, the genome sequences of certain pathogenic bacteria (Mycobacterium leprae, Yersinia pestis and Rickettsia prowazekii) show how an organism can undergo reductive evolution on a large scale (i.e. the dying out of families) as a result of niche change. There appears to be less pressure to delete pseudogenes in eukaryotes. These can be divided into two varieties, duplicated and processed, where the latter involves reverse transcription from an mRNA intermediate. We discuss these collectively in yeast, worm, fly, and human. The fly has few pseudogenes apparently because of its high rate of genomic DNA deletion. In the other three organisms, the distribution of pseudogenes on the chromosome and amongst different families is highly non-uniform. Pseudogenes tend not to occur in the middle of chromosome arms, and tend to be associated with lineage-specific (as opposed to highly conserved) families that have environmental-response functions. This may be because, rather than being dead, they may form a reservoir of diverse "extra parts" that can be resurrected to help an organism adapt to its surroundings. In yeast, there may be a novel mechanism involving the [PSI+] prion that potentially enables this resurrection. In worm, the pseudogenes tend to arise out of families (e.g. chemoreceptors) that are greatly expanded in it compared to the fly. The human genome stands out in having many processed pseudogenes. These have a character very different from those of the duplicated variety, to a large extent just representing random insertions. Thus, their occurrence tends to be roughly in proportion to the amount of mRNA for a particular protein and to reflect the extent of the intergenic sequences. Further information about pseudogenes is available at http://genecensus.org/pseudogene PMID- 12083510 TI - Evidence that the KH RNA-binding domains influence the action of the E. coli NusA protein. AB - The NusA transcription elongation protein, which binds RNA, contains sequences corresponding to the S1 and KH classes of identified RNA binding domains. An essential function in E. coli, NusA is also one of the host factors required for action of the N transcription antitermination protein of lambda. Tandem KH domains have been identified downstream of the S1 domain. We changed the first Gly to Asp of the GXXG motif, a tetrapeptide diagnostic of KH domains, of both NusA KH domains. The change in the first, G253D, has a large effect, while the change in the second, G319D, has a small effect on NusA action. The changes in both KH domains interfere with NusA binding to RNA. A change of a highly conserved Arg in the S1 domain, R199A, has previously been reported to interfere with RNA binding while exerting a small effect on NusA action. However, a nusA allele with both the R199A and G319D changes encodes a functionally inactive NusA protein. These studies provide direct evidence that the both KH as well as the S1 RNA binding domains are important for NusA action in support of bacterial viability as well as transcription antitermination mediated by the lambda N protein. PMID- 12083511 TI - Identification of new poly(A) polymerase-inhibitory proteins capable of regulating pre-mRNA polyadenylation. AB - The 3' ends of nearly all eukaryotic pre-mRNAs undergo cleavage and polyadenylation, thereby acquiring a poly(A) tail added by the enzyme poly(A) polymerase (PAP). Two well-characterized examples of regulated poly(A) tail addition in the nucleus consist of spliceosomal proteins, either the U1A or U170K proteins, binding to the pre-mRNA and inhibiting PAP via their PAP regulatory domains (PRDs). These two proteins are the only known examples of this type of gene regulation. On the basis of sequence comparisons, it was predicted that many other proteins, including some members of the SR family of splicing proteins, contain functional PRDs. Here we demonstrate that the putative PRDs found in the SR domains of the SR proteins SRP75 and U2AF65, via fusion to a heterologous MS2 RNA binding protein, specifically and efficiently inhibit PAP in vitro and pre mRNA polyadenylation in vitro and in vivo. A similar region from the SR domain of SRP40 does not exhibit these activities, indicating that this is not a general property of SR domains. We find that the polyadenylation- and PAP-inhibitory activity of a given polypeptide can be accurately predicted based on sequence similarity to known PRDs and can be measured even if the polypeptides' RNA target is unknown. Our results also indicate that PRDs function as part of a network of interactions within the pre-mRNA processing complex and suggest that this type of regulation will be more widespread than previously thought. PMID- 12083512 TI - Recognition of tRNA backbone for aminoacylation with cysteine: evolution from Escherichia coli to human. AB - The underlying basis of the genetic code is specific aminoacylation of tRNAs by aminoacyl-tRNA synthetases. Although the code is conserved, bases in tRNA that establish aminoacylation are not necessarily conserved. Even when the bases are conserved, positions of backbone groups that contribute to aminoacylation may vary. We show here that, although the Escherichia coli and human cysteinyl-tRNA synthetases both recognize the same bases (U73 and the GCA anticodon) of tRNA for aminoacylation, they have different emphasis on the tRNA backbone. The E. coli enzyme recognizes two clusters of phosphate groups. One is at A36 in the anticodon and the other is in the core of the tRNA structure and includes phosphate groups at positions 9, 12, 14, and 60. Metal-ion rescue experiments show that those at positions 9, 12, and 60 are involved with binding divalent metal ions that are important for aminoacylation. The E. coli enzyme also recognizes 2'-hydroxyl groups within the same two clusters: at positions 33, 35, and 36 in the anticodon loop, and at positions 49, 55, and 61 in the core. The human enzyme, by contrast, recognizes few phosphate or 2'-hydroxy groups for aminoacylation. The evolution from the backbone-dependent recognition by the E. coli enzyme to the backbone-independent recognition by the human enzyme demonstrates a previously unrecognized shift that nonetheless has preserved the specificity for aminoacylation with cysteine. PMID- 12083513 TI - Structure-function analysis of the inverted terminal repeats of the sleeping beauty transposon. AB - Translocation of Sleeping Beauty (SB) transposon requires specific binding of SB transposase to inverted terminal repeats (ITRs) of about 230 bp at each end of the transposon, which is followed by a cut-and-paste transfer of the transposon into a target DNA sequence. The ITRs contain two imperfect direct repeats (DRs) of about 32 bp. The outer DRs are at the extreme ends of the transposon whereas the inner DRs are located inside the transposon, 165-166 bp from the outer DRs. Here we investigated the roles of the DR elements in transposition. Although there is a core transposase-binding sequence common to all of the DRs, additional adjacent sequences are required for transposition and these sequences vary in the different DRs. As a result, SB transposase binds less tightly to the outer DRs than to the inner DRs. Two DRs are required in each ITR for transposition but they are not interchangeable for efficient transposition. Each DR appears to have a distinctive role in transposition. The spacing and sequence between the DR elements in an ITR affect transposition rates, suggesting a constrained geometry is involved in the interactions of SB transposase molecules in order to achieve precise mobilization. Transposons are flanked by TA dinucleotide base-pairs that are important for excision; elimination of the TA motif on one side of the transposon significantly reduces transposition while loss of TAs on both flanks of the transposon abolishes transposition. These findings have led to the construction of a more advanced transposon that should be useful in gene transfer and insertional mutagenesis in vertebrates. PMID- 12083514 TI - Structural mimicry of proline kinks: tertiary packing interactions support local structural distortions. AB - Proline residues in the helical segments of soluble and transmembrane proteins have received special attention from both a structural and functional perspective. A feature of these helices is the structural distortion termed "proline-kink", which has been associated with the presence of the proline residue. However, a recent report on the yeast heat-shock transcription factor of Kluyveromyces lactis (HSF_KL) suggests that these proline-associated deformations can be achieved in the absence of proline residues, thus raising the question of the mechanisms responsible for the structural mimicry of proline-related features. In this study, the specific interactions responsible for the distortion were characterized by comparative analysis of the atomic details of the packing interactions that surround the evolutionarily conserved proline-kink in the alpha2 helix of HSF_KL and a set of 39 structurally related proteins that lacked the distortion. The mechanistic details inferred from this analysis were confirmed with molecular dynamics simulations. The study shows that the packing interactions between the alpha2 and alpha1 helices in HSF_KL are responsible for the stabilization of the conserved kink, whether a proline residue that divides the helix into segments is present or not. The proline-kink can facilitate the formation of tertiary packing interactions that would otherwise not be possible. However, it is the ability to establish differential packing interactions for the helix segments, rather than the structural properties of the proline-kink itself, that emerges as the key factor for the characteristic distortion. PMID- 12083515 TI - Biochemical properties of single-stranded DNA-binding protein from Mycobacterium smegmatis, a fast-growing mycobacterium and its physical and functional interaction with uracil DNA glycosylases. AB - The single-stranded DNA-binding proteins (SSBs) are vital to virtually all DNA functions. Here, we report on the biochemical properties of SSB from a fast growing mycobacteria, Mycobacterium smegmatis, and the interaction of the homotetrameric SSBs with uracil DNA glycosylases (UDGs) from M. smegmatis (Msm), Mycobacterium tuberculosis (Mtu) and Escherichia coli (Eco). UDG is a crucial DNA repair enzyme, which removes the promutagenic uracil residues. MsmSSB stimulates activity of the homologous Msm UDG and of the heterologous Mtu-, and Eco-UDGs. On the contrary, while the MtuSSB stimulates the Mtu UDG, it inhibits the other two UDGs. Although the MsmSSB shares 84% identity with MtuSSB, the two are strikingly different, in that MsmSSB contains a glycine-rich segment (11 out of 13 residues) in the spacer connecting the N-terminal DNA-binding domain with the C-terminal acidic tail. While the DNA-binding properties of MsmSSB, such as its affinity to oligomeric DNA, requirement of minimum size DNA and the modes of interaction are indistinguishable from those of Eco-, and Mtu-SSBs, it is unclear if the glycine rich segment confers structural advantage to MsmSSB, responsible for its stimulatory effect on all UDGs tested. More importantly, by using a small polypeptide inhibitor of UDGs, and the deletion mutants of SSBs, we suggest that the C-terminal acidic tail of the SSBs interacts within the DNA-binding groove of the UDGs, and propose a role for SSBs in the recruitment of UDGs to the damaged DNA. PMID- 12083516 TI - The 1.9 A crystal structure of heat-labile shrimp alkaline phosphatase. AB - Alkaline phosphatases are non-specific phosphomonoesterases that are distributed widely in species ranging from bacteria to man. This study has concentrated on the tissue-nonspecific alkaline phosphatase from arctic shrimps (shrimp alkaline phosphatase, SAP). Originating from a cold-active species, SAP is thermolabile and is used widely in vitro, e.g. to dephosphorylate DNA or dNTPs, since it can be inactivated by a short rise in temperature. Since alkaline phosphatases are zinc-containing enzymes, a multiwavelength anomalous dispersion (MAD) experiment was performed on the zinc K edge, which led to the determination of the structure to a resolution of 1.9 A. Anomalous data clearly showed the presence of a zinc triad in the active site, whereas alkaline phosphatases usually contain two zinc and one magnesium ion per monomer. SAP shares the core, an extended beta-sheet flanked by alpha-helices, and a metal triad with the currently known alkaline phosphatase structures (Escherichia coli structures and a human placental structure). Although SAP lacks some features specific for the mammalian enzyme, their backbones are very similar and may therefore be typical for other higher organisms. Furthermore, SAP possesses a striking feature that the other structures lack: surface potential representations show that the enzyme's net charge of -80 is distributed such that the surface is predominantly negatively charged, except for the positively charged active site. The negatively charged substrate must therefore be directed strongly towards the active site. It is generally accepted that optimization of the electrostatics is one of the characteristics related to cold-adaptation. SAP demonstrates this principle very clearly. PMID- 12083517 TI - Orientation changes of the myosin light chain domain during filament sliding in active and rigor muscle. AB - Structural changes in myosin power many types of cell motility including muscle contraction. Tilting of the myosin light chain domain (LCD) seems to be the final step in transducing the energy of ATP hydrolysis, amplifying small structural changes near the ATP binding site into nanometer-scale motions of the filaments. Here we used polarized fluorescence measurements from bifunctional rhodamine probes attached at known orientations in the LCD to describe the distribution of orientations of the LCD in active contraction and rigor. We applied rapid length steps to perturb the orientations of the population of myosin heads that are attached to actin, and thereby characterized the motions of these force-bearing myosin heads. During active contraction, this population is a small fraction of the total. When the filaments slide in the shortening direction in active contraction, the long axis of LCD tilts towards its nucleotide-free orientation with no significant twisting around this axis. In contrast, filament sliding in rigor produces coordinated tilting and twisting motions. PMID- 12083518 TI - Crystal structure of a non-canonical low-affinity peptide complexed with MHC class I: a new approach for vaccine design. AB - Peptides bind with high affinity to MHC class I molecules by anchoring certain side-chains (anchors) into specificity pockets in the MHC peptide-binding groove. Peptides that do not contain these canonical anchor residues normally have low affinity, resulting in impaired pMHC stability and loss of immunogenicity. Here, we report the crystal structure at 1.6 A resolution of an immunogenic, low affinity peptide from the tumor-associated antigen MUC1, bound to H-2Kb. Stable binding is still achieved despite small, non-canonical residues in the C and F anchor pockets. This structure reveals how low-affinity peptides can be utilized in the design of novel peptide-based tumor vaccines. The molecular interactions elucidated in this non-canonical low-affinity peptide MHC complex should help uncover additional immunogenic peptides from primary protein sequences and aid in the design of alternative approaches for T-cell vaccines. PMID- 12083519 TI - Crystal structure of a non-canonical high affinity peptide complexed with MHC class I: a novel use of alternative anchors. AB - The crystal structure of a non-standard peptide, YEA9, in complex with H-2Kb, at 1.5 A resolution demonstrates how YEA9 peptide can bind with surprisingly high affinity through insertion of alternative, long, non-canonical anchors into the B and E pockets. The use of "alternative pockets" represents a new mode of high affinity peptide binding, that should be considered when predicting peptide epitopes for MHC class I. These novel interactions encountered in this non canonical high affinity peptide-MHC complex should help predict additional binding peptides from primary protein sequences and aid in the design of alternative approaches for peptide-based vaccines. PMID- 12083521 TI - Amino acid modification in the HIV-1 Tat basic domain: insights from molecular dynamics and in vivo functional studies. AB - Tat is an essential protein of the human immunodeficiency virus type 1 (HIV-1). It activates transcription by specifically binding a stem-loop element in the viral long terminal repeat through its highly basic arginine-rich domain. Conserved lysine residues at positions 50 and 51 inside this domain have been recently reported to be the targets of post-translational modification by acetylation, and mutation of these residues has pointed out its relevance to protein function. In an attempt to shed light on the molecular basis of the functional differences found for Tat mutants we have performed a series of molecular dynamics simulations on wt Tat, Lys50/51 --> Arg50/51, Lys50/51 --> Ala50/51 and acetylated Lys50 from HIV-1 strain Z2. Theoretical results are compared with a homogeneous set of in vivo transactivation assays on the corresponding Tat mutants from the strain B2, which exhibits high structural similarity with Tat from HIV-1 strain Z2. Remarkable correlation is found between the degree of structure conservation and the transactivation capabilities of Tat mutants. PMID- 12083520 TI - The structural basis of riboflavin binding to Schizosaccharomyces pombe 6,7 dimethyl-8-ribityllumazine synthase. AB - Riboflavin is an essential cofactor in all organisms. Its direct biosynthetic precursor, 6,7-dimethyl-8-ribityllumazine, is synthesised by the enzyme 6,7 dimethyl-8-ribityllumazine synthase. Recently, we have found that the enzyme from Schizosaccharomyces pombe binds riboflavin, the final product of the pathway with a relatively high affinity with a KD of 1.2 microM. Here, we report on the crystal structure of lumazine synthase from S. pombe with bound riboflavin and compare the binding mode with those of the substrate analogue inhibitor 5-nitro-6 (D-ribitylamino)-2,4(1H,3H)-pyrimidinedione and of the product analogue 6 carboxyethyl-7-oxo-8-ribityllumazine. In all complexes the pyrimidinedione moieties of each respective ligand bind in a very similar orientation. Binding of riboflavin additionally involves a stacking interaction of the dimethylbenzene moiety with the side-chain of His94, a highly conserved residue in all lumazine synthases. The enzyme from Bacillus subtilis showed a KD of at least 1 mM whereas the very homologous enzyme from Saccharomyces cerevisiae had a comparable KD of 3.9 microM. Structural comparison of the S. cerevisiae, the S. pombe, and the mutant enzymes suggests that fine tuning of affinity is achieved by influencing this stacking interaction. PMID- 12083522 TI - Origins of the high stability of an in vitro-selected cold-shock protein. AB - In previous work, we had identified stabilized forms of the cold-shock protein Bs CspB from Bacillus subtilis in a combinatorial library by an in vitro selection procedure. In this library, the sequence positions 2, 3, 46, 64, 66, and 67 had been randomized, because Bs-CspB differs from the naturally thermostable homolog Bc-Csp from Bacillus caldolyticus, among others, at these six positions. For the most stable selected variant, the midpoint of thermal unfolding (tM) increased by 28.2 deg. C and the Gibbs free energy of unfolding (deltaG(D)) by 19 kJ/mol. Here, we analyzed by site-directed mutagenesis how the selected residues contribute individually to this strong stabilization. Val3 and Val66, which replace Glu3 and Glu66 of wild-type Bs-CspB, each contribute about 7 kJ/mol to stability, the Thr64Arg substitution contributes 4.5 kJ/mol, and 3.2 kJ/mol originate from the Ala46Leu replacement. Gly67 at the carboxy terminus is unimportant for stability, the Arg selected at position 2 is overall slightly destabilizing but improves the coulombic interactions. The best variant differs from Bc-Csp at all six positions; nevertheless, natural and in vitro selection followed similar principles. In both cases, negatively charged residues at the adjacent positions 3 and 66 are avoided, and a positively charged residue is introduced into this area of the protein surface. Its exact location is unimportant. It can be at position 3, as in the thermophilic Bc-Csp, or at positions 2 or 64, as in the most stable selected variant. These positively charged residues contribute to stability not by engaging in pairwise coulombic interactions with a specific carboxyl group, but by generally improving the charge distribution in this particular region of the protein surface. These coulombic effects contribute significantly to the thermostability of the cold shock proteins. They are only weakly interdependent and best explained by the presence of a flexible ion network at the protein surface. Our results emphasize that surface positions are very good candidates for optimizing protein stability. PMID- 12083523 TI - Two distinct stress signaling pathways converge upon the CHOP promoter during the mammalian unfolded protein response. AB - CHOP is a non-ER localized transcription factor that is induced by a variety of adverse physiological conditions including ER stress. Accumulation of unfolded proteins in the ER activates an unfolded protein response pathway that targets both ER resident chaperones (e.g. BiP) and CHOP. Hence, it is unclear if CHOP induction during ER stress occurs through the ER stress response element that is conserved in both CHOP and ER chaperone promoters, or through a separate regulatory pathway conserved among different CHOP inducing cellular stress conditions. We identified a bona fide ER stress element in the hamster CHOP promoter and found that similar transcription complexes containing NF-Y bound to both the CHOP and BiP ER stress response elements. In addition, we demonstrated for the first time the importance of the C/EBP-ATF composite site for CHOP regulation during ER stress. Activation of the ER transmembrane eIF2alpha kinase, PERK, induced ATF4 protein expression, direct binding to the composite site in CHOP promoter, and as a consequence, CHOP protein induction. We propose that this eIF2alpha-kinase/ATF4/C/EBP-ATF composite site pathway is conserved for CHOP regulation during various cellular stress conditions including ER stress. Our data indicate that both the ERSE and the PERK-ATF4 pathways converge on the CHOP promoter during ER stress and provide insights into the similarities and differences between CHOP and ER chaperone expression during normal and stress conditions. PMID- 12083524 TI - Crystal structure of the CCTgamma apical domain: implications for substrate binding to the eukaryotic cytosolic chaperonin. AB - The chaperonin containing TCP-1 (CCT, also known as TRiC) is the only member of the chaperonin family found in the cytosol of eukaryotes. Like other chaperonins, it assists the folding of newly synthesised proteins. It is, however, unique in its specificity towards only a small subset of non-native proteins. We determined two crystal structures of mouse CCTgamma apical domain at 2.2 A and 2.8 A resolution. They reveal a surface patch facing the inside of the torus that is highly evolutionarily conserved and specific for the CCTgamma apical domain. This putative substrate-binding region consists of predominantly positively charged side-chains. It suggests that the specificity of this apical domain towards its substrate, partially folded tubulin, is conferred by polar and electrostatic interactions. The site and nature of substrate interaction are thus profoundly different between CCT and its eubacterial homologue GroEL, consistent with their different functions in general versus specific protein folding assistance. PMID- 12083525 TI - Identification and structure of a putative Ca2+-binding domain at the C terminus of AQP1. AB - Aquaporin-1 (AQP1) is the first functionally identified aquaporin of a growing family of membrane water channels found in all forms of life. Recently, a possible secondary function as a cyclic guanosine monophosphate (cGMP) gated ion channel was attributed to AQP1. We have reconstituted purified protein from bovine and human red blood cell membranes into highly ordered 2D crystals. The topography of both AQP1s was determined by electron microscopy from freeze-dried, unidirectionally metal-shadowed 2D crystals as well as from surface topographs of native crystals recorded in buffer solution with the atomic force microscope (AFM). In spite of the high level of sequence homology between bovine and human AQP1, the surfaces showed distinct differences. Alignment of both sequences and comparison of the acquired surface topographies with the atomic model of human AQP1 revealed the topographic changes on the surface of bovine AQP1 to be induced by a few amino acid substitutions. A striking degree of sequence homology was found between the carboxyl-terminal domains of AQP1s from different organisms and EF-hands from Ca2+-binding proteins belonging to the calmodulin superfamily, suggesting the existence of a Ca2+-binding site at the C terminus of AQP1 instead of the putative cGMP-binding site reported previously. To unveil its position on the acquired surface topographies, 2D crystals of AQP1 were digested with carboxypeptidase Y, which cleaves off the intracellular C terminus. Difference maps of AFM topographs between the native and the peptidase-treated AQP1s showed the carboxylic tail to be close to the 4-fold symmetry axis of the tetramer. SDS PAGE and matrix-assisted laser desorption/ionisation mass spectrometry of native and decarboxylated bovine and human AQP1 revealed that the EF-hand motif found at the C terminus of AQP1 was partially resistant to peptidase digestion. The importance of the C-terminal domain is implicated by structural instability of decarboxylated AQP1. A possible role of the C terminus and calcium in translocation of AQP1 in cholangiocytes from intracellular vesicles to the plasma membrane and in triggering its fusion is discussed. Functional studies are now required to identify the physiological role of the Ca2+-binding site. PMID- 12083526 TI - The solution structure of the bacteriophage lambda head-tail joining protein, gpFII. AB - The bacteriophage lambda FII protein (gpFII) is a 117 residue structural protein found in the phage particle that is required for the joining of phage heads and tails at the last step of morphogenesis. We have performed biophysical experiments to show that gpFII is stable, monomeric, and reversibly folded. We have also determined the atomic resolution structure of gpFII using NMR spectroscopy. gpFII is shown to possess a novel fold consisting of seven beta strands and a short alpha-helix. It also displays two large unstructured regions at the N terminus (residues 1-24) and in a large loop near the middle of the protein (residues 46-62). We speculate that these unstructured regions become structured when gpFII assembles into the phage particle, and that these conformational changes play an important role in regulating the assembly pathway. Alignment of the gpFII sequence with those of homologues from other lambdoid phages has allowed us to putatively identify distinct surfaces on the gpFII structure that mediate binding to the phage head and tail. PMID- 12083527 TI - Five atomic resolution structures of endothiapepsin inhibitor complexes: implications for the aspartic proteinase mechanism. AB - Endothiapepsin is derived from the fungus Endothia parasitica and is a member of the aspartic proteinase class of enzymes. This class of enzyme is comprised of two structurally similar lobes, each lobe contributing an aspartic acid residue to form a catalytic dyad that acts to cleave the substrate peptide bond. The three-dimensional structures of endothiapepsin bound to five transition state analogue inhibitors (H189, H256, CP-80,794, PD-129,541 and PD-130,328) have been solved at atomic resolution allowing full anisotropic modelling of each complex. The active sites of the five structures have been studied with a view to studying the catalytic mechanism of the aspartic proteinases by locating the active site protons by carboxyl bond length differences and electron density analysis. In the CP-80,794 structure there is excellent electron density for the hydrogen on the inhibitory statine hydroxyl group which forms a hydrogen bond with the inner oxygen of Asp32. The location of this proton has implications for the catalytic mechanism of the aspartic proteinases as it is consistent with the proposed mechanism in which Asp32 is the negatively charged aspartate. A number of short hydrogen bonds (approximately 2.6 A) with ESD values of around 0.01 A that may have a role in catalysis have been identified within the active site of each structure; the lengths of these bonds have been confirmed using NMR techniques. The possibility and implications of low barrier hydrogen bonds in the active site are considered. PMID- 12083528 TI - The 3.0 A resolution crystal structure of glycosomal pyruvate phosphate dikinase from Trypanosoma brucei. AB - The crystal structure of the glycosomal enzyme pyruvate phosphate dikinase from the African protozoan parasite Trypanosoma brucei has been solved to 3.0 A resolution by molecular replacement. The search model was the 2.3 A resolution structure of the Clostridium symbiosum enzyme. Due to different relative orientations of the domains and sub-domains in the two structures, molecular replacement could be achieved only by positioning these elements (four bodies altogether) sequentially in the asymmetric unit of the P2(1)2(1)2 crystal, which contains one pyruvate phosphate dikinase (PPDK) subunit. The refined model, comprising 898 residues and 188 solvent molecules per subunit, has a crystallographic residual index Rf = 0.245 (cross-validation residual index Rfree = 0.291) and displays satisfactory stereochemistry. Eight regions, comprising a total of 69 amino acid residues at the surface of the molecule, are disordered in this crystal form. The PPDK subunits are arranged around the crystallographic 2 fold axis as a dimer, analogous to that observed in the C. symbiosum enzyme. Comparison of the two structures was carried out by superposition of the models. Although the fold of each domain or sub-domain is similar, the relative orientations of these constitutive elements are different in the two structures. The trypanosome enzyme is more "bent" than the bacterial enzyme, with bending increasing from the center of the molecule (close to the molecular 2-fold axis) towards the periphery where the N-terminal domain is located. As a consequence of this increased bending and of the differences in relative positions of subdomains, the nucleotide-binding cleft in the amino-terminal domain is wider in T. brucei PPDK: the N-terminal fragment of the amino-terminal domain is distant from the catalytic, phospho-transfer competent histidine 482 (ca 10 A away). Our observations suggest that the requirements of domain motion during enzyme catalysis might include widening of the nucleotide-binding cleft to allow access and departure of the AMP or ATP ligand. PMID- 12083529 TI - The modulation of androgen metabolism by estradiol, minocycline, and indomethacin in a cell culture model. AB - BACKGROUND: This investigation attempts to clarify the proanabolic effects of minocycline and indomethacin by studying their effects on androgen metabolism and mediation by estradiol. A cell culture model was used with androgen substrates because of the proanabolic effects of androgen metabolites. METHODS: Monolayer cultures of human gingival fibroblasts (HGF) derived from 6 patients were incubated in duplicate with 14C- testosterone or 14C-4-androstenedione as substrates and optimal concentrations of estradiol (E1,3 microgram/ml) and minocycline (M25 microgram/ml) or indomethacin (I, 1 microgram/ml) alone and in combination (E1,3+11 or E1,3+M25 microgram/ml); similar experiments were carried out with human oral periosteal fibroblasts (HPF), M, I, E, and the combinations. At the end of a 24-hour incubation period in Eagle's MEM, the medium was solvent extracted with ethyl acetate and the metabolites were separated by TLC in a benzene:acetone solvent system (4:1 v/v). The separated metabolites were quantified using a radioisotope scanner. RESULTS: Both androgens were metabolized to 5alpha-dihydrotestosterone (DHT) and 4-androstenedione (4-A) or testosterone (T) at baseline and in response to the agents tested, by HGF and HPF. With HGF, there were significant increases in the yields of DHT and 4-A or T in response to M, E, and M+E, resulting in 50% to 2.4-fold increases in these metabolites over control incubations (n = 6; P<0.01). The responses to I and combinations of I+E were similar. HPF also demonstrated significant increases of 29% to 4-fold in the yields of androgen metabolites in response to M, E, and M+E (n = 6; P<0.01). I and E similarly increased the yields of androgen metabolites, alone and in combination. CONCLUSIONS: Adjunctive periodontal treatment with minocycline or indomethacin can contribute to hormone-modulated anabolic responses in males and females in gingival and periosteal fibroblasts derived from a chronically inflamed source. PMID- 12083531 TI - Subgingival calculus detection with fluorescence induced by 655 nm InGaAsP diode laser radiation. AB - BACKGROUND: It has been suggested that the 655 nm InGaAsP diode laser radiation can induce considerably higher amounts of fluorescence in subgingival calculus than in cementum. This in vitro study evaluated the use of fluorescence induced by the diode laser radiation for the detection of subgingival calculus. METHODS: The study sample consisted of 30 extracted molars, premolars, canines, and incisors. The root surface of each sample was partially covered with subgingival calculus. The source of laser radiation was an InGaAsP diode laser emitting visible radiation at a wavelength of 655 nm. The fluorescent radiation reflected from the cementum and calculus was detected by a photoelectric cell integrated within the diagnostic system. The samples were randomly assigned to 3 experimental groups of 10 teeth each for detection of calculus in various media. Specifically, the fluorescence was determined in air (group A), in electrolytic salt solution (group B), and in blood (group C). For each sample, the intensity of the fluorescent light was evaluated separately for cementum and calculus in triplicate measurements. The statistical analysis was performed using a t test at a level of significance of 5% (P<0.05). RESULTS: The intensities of fluorescence radiation emitted by cementum were 0.4 (+/-0.51) in air, 0.4 (+/-0.51) in electrolytic solution, and 2.13 (+/-0.59) in blood. For calculus, the relative intensities of fluorescence radiation were 54.1 (+/-29.09) in air, 60.77 (+/ 28.35) in electrolytic salt solution, and 39.63 (+/-29.79) in blood. The differences in fluorescence between cementum and calculus within each experimental unit were significant (group A: P= 0.0002, group B: P= 0.0001, group C: P= 0.031). The intensity of fluorescence in calculus was also significantly different between the 3 experimental groups. CONCLUSION: The present study indicates that subgingival calculus can be reliably detected on extracted teeth using laser fluorescence induced by the 655 nm diode-laser radiation. In vivo studies are necessary to evaluate the clinical utility of this procedure. PMID- 12083532 TI - Relationship between compliance and periodontal treatment outcome in smokers. AB - BACKGROUND: Smoking is an established risk factor of periodontal disease and smokers are regarded as patients with a high risk of periodontitis recurrence during the maintenance phase. Lack of compliance and smoking constitute significant factors for the risk of further periodontitis progression. The purpose of the present study was to investigate the relationship between periodontal status and the tendency to interrupt periodontal treatment and determine if this relationship differs significantly between smokers and non smokers. METHODS: The investigation was conducted as a retrospective study on a sample of 325 patients referred for treatment. The patients had been offered full periodontal treatment and a full-mouth oral radiographic examination. In order to investigate any correlations between periodontal status and smoking or interrupted periodontal treatments, stepwise multiple regression analyses were adopted. RESULTS: The mean age of the sample was 49.7 years (range 25 to 83) and a majority were females (57%). The relative frequency of smoking was 52%. The relative frequency of interruption of periodontal treatment was 26% for non smokers and 31% for smokers. Smokers who interrupted periodontal treatment after the reevaluation were found to have significantly deeper periodontal probing depths at the reevaluation compared to those who did not interrupt the treatment irrespective of smoking habits (P<0.001). CONCLUSIONS: The results of the study demonstrate that the non-complying individuals had the highest risk of recurrent periodontitis even if they had completed the treatment plan. An important task in the future will be to find ways to reduce the frequency of non-compliance and thus improve the prognosis. PMID- 12083530 TI - Genetic dependence of the specific T-cell cytokine response to Porphyromonas gingivalis in mice. AB - BACKGROUND: Susceptibility to periodontal infections may, in part, be genetically determined. Porphyromonas gingivalis is a major periodontopathogen, and the immune response to this organism requires T-cell help. The aim of the present study was to examine the specific T-cell cytokine responses to P. gingivalis outer membrane antigens in a mouse model and their relationship with H-2 haplotype. METHODS: BALB/c and DBA/2J (H-2d), CBACaH (H-2k), and C57BL6 (H-2b) mice were immunized with P. gingivalis outer membrane antigens weekly for 3 weeks. One week after the final injection, the spleens were removed, and 6 T-cell lines specific for P. gingivalis were established for each mouse strain. The percentage of CD4 and CD8 cells in the P. gingivalis-specific T-cell lines staining positive for intracytoplasmic interleukin (IL)-4, interferon (IFN) gamma, and IL-10 was determined by 2-color flow cytometry. RESULTS: The cytokine profiles of T-cell lines from BALB/c and DBA/2J mice showed no significant differences. Significantly fewer IL-4+, IFN-gamma+, and IL-10+ CD4 cells than IL 4+, IFN-gamma+, and IL-10+ CD8 cells, respectively, were demonstrated for both strains. P. gingivalis-specific T-cell lines generated from CBACaH mice were similar to those generated from BALB/c and DBA/2J mice; however, the mean percentage of IL-4+ CD4 cells in CBACaH mice was lower than the percentage of IFN gamma+ CD4 cells. Also, the mean percentage of IFN-gamma+ CD4 cells in CBACaH mice was significantly increased compared to DBA/2J mice. Unlike the other 3 strains, T-cell lines established from C57BL6 mice contained similar percentages of cytokine-positive cells, although the percentage of IL-4+ CD4 cells was reduced in comparison to the percentage of CD8 cells. However, comparisons with the other 3 strains demonstrated a higher percentage of IL-4+ CD4 cells than in lines established from the spleens of DBA/2J mice, IFN-gamma+ CD4 cells than in lines established from BALB/c and CBACaH mice, and IL-10+ CD4 cells than in lines established from all 3 other strains. No significant differences in the percentage of positive CD8 cells were demonstrated between lines in the 4 strains of mice. CONCLUSION: The specific T-cell response to P. gingivalis in mice may, in the case of the CD4 response, depend on MHC genes. These findings are consistent with the concept that patient susceptibility is important to the outcome of periodontal infection and may, in part, be genetically determined. PMID- 12083533 TI - The effect of subgingival controlled-release delivery of chlorhexidine chip on clinical parameters and matrix metalloproteinase-8 levels in gingival crevicular fluid. AB - BACKGROUND: The present study evaluated the efficacy of controlled-release delivery of chlorhexidine gluconate (CHX) on clinical parameters and on gingival crevicular fluid (GCF) matrix metalloproteinase (MMP)-8 levels in chronic periodontitis patients. METHODS: Twenty patients with chronic periodontitis were screened for 6 months. Two interproximal sites were selected from mesial surfaces of anterior teeth with probing depths of 6 to 8 mm that bled on probing in each patient. There were at least 2 teeth between the selected sites. CHX chip was inserted into a randomly selected site following scaling and root planing (SRP+CHX), while the other selected site received only SRP in each patient. Probing depth (PD), clinical attachment level (CAL), plaque index (PI), and papilla bleeding index (PBI) were recorded at baseline and at 1, 3, and 6 months. GCF MMP-8 levels were analyzed at baseline; 2 and 10 days; and at 1, 3, and 6 months by immunofluorometric assay (IFMA). RESULTS: At baseline, there were no statistically significant differences in the mean PD, CAL, PBI, and PI scores between SRP+CHX and SRP alone groups. At 1, 3, and 6 months, all clinical parameters in each group significantly decreased (P <0.0167) when compared to baseline. The reduction of PD and improvement in CAL were higher in the SRP+CHX group compared to SRP alone at 3 and 6 months. However, the differences between the 2 groups were not statistically significant. PBI and PI scores were not significantly different between SRP+CHX and SRP alone groups at any visit. GCF MMP-8 levels were similar in both groups at baseline. Intragroup analysis showed significant decreases in the GCF MMP-8 level for the SRP+CHX group between baseline and 1, 3, and 6 months (P<0.01). Intergroup analysis demonstrated significantly lower mean levels of GCF MMP-8 at 1 month in the SRP+CHX group compared to the SRP alone group (P <0.05). CONCLUSIONS: These data suggest that CHX chip application following SRP is beneficial in improving periodontal parameters and reducing GCF MMP-8 levels for 6 months' duration. The use of a chairside MMP-8 dipstick periodontitis test might be a useful adjunctive diagnostic tool when monitoring the course of CHX chip treatment. PMID- 12083534 TI - Lactoferrin iron levels affect attachment of Actinobacillus actinomycetemcomitans to buccal epithelial cells. AB - BACKGROUND: Prior reports have suggested that the iron-binding protein lactoferrin (LF) may either kill Actinobacillus actinomycetemcomitans (Aa) or interfere with its binding to host cells. Other studies have indicated that the degree of iron saturation of LF might play a role in these interactions. However, these studies utilized strains that had lost critical attachment characteristics found in well-preserved clinical isolates of Aa. The purpose of this work was to study the effect of LF iron levels on survival and attachment of well-preserved clinical isolates of Aa. METHODS: LF containing 0%, 30%, and 100% iron saturation was tested for its ability to kill clinical isolates of Aa and to inhibit their binding to buccal epithelial cells (BECs). RESULTS: Neither iron-free LF (apo-LF) nor iron-saturated LF killed Aa clinical isolates. Increasing the iron saturation of LF resulted in an increased inhibition of Aa binding to BECs (P < or =0.005). This effect was consistent for the 3 clinical isolates tested. Pretreatment of Aa with iron-saturated LF reduced binding to BECs by 58%, 61.8%, and 64.2%, respectively, for each of the 3 clinical strains tested (P < or =0.005). Pretreatment of Aa strains with apo-LF, iron alone, or bovine serum albumin had no effect on binding. Pretreatment of BECs with LF (either apo-LF or iron containing LF) had no influence on Aa binding. CONCLUSIONS: These results indicate that reduction in binding of Aa to epithelial cells is maximized by pretreatment of Aa cells with iron-saturated lactoferrin. These in vitro results suggest that patients with lactoferrin containing lowered levels of iron would be more susceptible to Aa colonization. PMID- 12083535 TI - Lactoferrin iron levels are reduced in saliva of patients with localized aggressive periodontitis. AB - BACKGROUND: Actinobacillus actinomycetemcomitans (Aa) is associated with localized aggressive periodontal disease in juveniles (LAgP). Lactoferrin (LF) is an iron-binding salivary protein that has been shown to kill Aa in its iron-free form (apo) and reduce binding to host cells in its iron-saturated form (halo). However, recent in vitro studies show that LF does not kill clinical isolates of Aa, and LF with reduced levels of bound iron does not interfere with its attachment. These findings suggest that colonization of Aa may occur more readily in an environment containing LF with low iron levels. The purpose of this study was to examine the relationship of LF iron levels in saliva of LAgP patients as compared to their age-, gender-, and race-matched controls. METHODS: Whole and parotid saliva was collected from LAgP patients and matched controls. Micrograms of LF/mg of protein as well as nanograms of iron/micrograms of LF were determined. Iron binding was determined in parotid saliva by addition of nonlabeled and 59Fe labeled iron. RESULTS: LAgP patients' whole saliva had higher LF levels than controls, but their LF contained less iron (P < or =0.005). No iron was found in LF from parotid saliva in either group. When iron was added to parotid saliva, the LAgP saliva bound 20 to 30 times less iron than controls (P< or =0.001). Finally, LF was identified as the major iron-binding protein in parotid saliva by 59Fe autoradiography and Western blotting. CONCLUSIONS: This study shows that the level of bound iron in LF is significantly reduced in LAgP patients compared to controls. These data suggest that LF from LAgP patients has a reduced capacity to bind iron and that LF iron levels may play an important role in Aa-induced LAgP. PMID- 12083536 TI - Periodontal assessment of patients undergoing angioplasty for treatment of coronary artery disease. AB - BACKGROUND: The purpose of this cross-sectional study was to assess the periodontal condition of patients with coronary artery disease (CAD) in order to verify the association between CAD and certain periodontal parameters. METHODS: Eighty patients (48 males and 32 females) were recruited from the Cleveland University Hospitals, Division of Cardiology (aged 23 to 83, median age 54 years). Upon cardiac catheterization, 50 were diagnosed with severe CAD (experimental group) and 30 with no angiographic evidence of CAD (control group). Patients with CAD were divided into 3 subgroups according to the clinical diagnosis of acute infarction (Al) (n = 20), stable angina (SA) (n = 20), and unstable angina (UA) (n = 10). The following dental, medical, and social histories were recorded: number of dental visits/year, frequency of brushing/day and flossing/week, tobacco use, presence/absence of hypertension, diabetes, total cholesterol level, and the highest level of education completed. The following periodontal parameters were also assessed for each subject: plaque index (PI), gingival index (GI), bleeding on probing (BOP), probing depth (PD), clinical attachment level (CAL), and number of missing teeth. Logistic regression analysis was used to determine whether there was a significant correlation between these periodontal parameters and CAD. RESULTS: The significant confounding factors for the present study were found to be age and gender. The periodontal condition of the 80 subjects was clinically diagnosed as gingivitis and/or mild periodontitis. Analysis of the data after adjusting for age and gender demonstrated a statistically significant association between BOP and GI and CAD in patients with Al; and facial/lingual CAL in patients with UA. No significant associations were found between PD, interproximal CAL, number of missing teeth, or dental/medical histories and CAD. CONCLUSION: Based on the results of this study, gingival inflammation may be considered a more significant risk factor for CAD than previously reported. PMID- 12083537 TI - Probing bone level measurements for determination of the depths of Class II furcation defects. AB - BACKGROUND: Probing bone measurements as an alternative to open bone measurements to evaluate regenerative procedures in furcation defects do not seem to be used as yet. The purpose of the present study was to investigate the reliability of probing bone measurements in such defects. METHODS: Fifteen patients scheduled for surgical treatment of a total of 30 mandibular molars with buccal or lingual Class II furcation defects were studied. During treatment, duplicate vertical and horizontal recordings of probing attachment levels, probing bone levels, and open bone levels were taken by independent examiners. RESULTS: Deviations of both vertical and horizontal recordings between the first and second examiners were within +/-1 mm in 90% to 100% of examined sites for all 3 measurements. Standard deviations of differences between pairs of duplicate recordings were also similar for the 3 measurements both for vertical and horizontal defect dimensions and amounted to 0.7 to 0.9 mm. On average, vertical and horizontal open bone levels were 0.9 to 1.1 mm deeper than probing bone levels. Probing bone levels, in turn, were 1.1 to 1.5 mm deeper than probing attachment levels. CONCLUSIONS: In view of the consistency demonstrated between probing bone level and open bone level measurements in mandibular Class II defects, coupled with the additional discomfort for the patient of a reentry surgery and a possible reentry traumatic effect, open bone level measurements do not seem necessary or even justified to evaluate effects of periodontal therapy in these defects, and can be substituted by probing bone measurements. PMID- 12083538 TI - Scanning electron microscopy and Fourier transformed infrared spectroscopy analysis of bone removal using Er:YAG and CO2 lasers. AB - BACKGROUND: A thorough analysis of laser-ablated bone tissue is required before applying the technique to osseous surgery. In this study, we examine the morphological features and chemical composition of the bone surface after Er:YAG and CO2 lasers ablation. METHODS: Six Wistar rats were used. An Er:YAG laser was used for ablation at an output energy of 100 mJ/pulse and a pulse rate of 10 Hz (1 W). Continuous CO2 laser irradiation was performed at an output energy of 1 W. Sites drilled using a conventional micromotor were used as controls. Analysis using scanning electron microscopy (SEM) and Fourier transformed infrared (FTIR) spectroscopy was performed. RESULTS: Er:YAG laser ablation produced a groove with similar dimensions to that produced by bur drilling, whereas the CO2 laser produced only a charred line with minimal tissue removal. SEM observations revealed that the groove produced by the Er:YAG laser had well-defined edges and a smear layer-free surface with a characteristically rough appearance and with entrapped fibrin-like tissue. The melting and carbonization produced by the CO2 laser were not observed on sites irradiated by the Er:YAG laser. FTIR spectroscopy revealed that the chemical composition of the bone surface after Er:YAG laser ablation was much the same as that following bur drilling. The production of toxic substances that occurred after CO2 laser irradiation was not observed following Er:YAG laser irradiation or bur drilling. CONCLUSION: These results suggest that the use of Er:YAG laser ablation may become an alternative method for oral and periodontal osseous surgery. PMID- 12083539 TI - Squamous odontogenic tumor: diagnosis and management. AB - The squamous odontogenic tumor (SOT) is a rare, benign, locally infiltrative neoplasm of the jaws that appears to originate from the rests of Malassez. It has been confused with other pathologic entities such as ameloblastomas, carcinomas, and fibromas and clinically may resemble localized periodontal disease. The tumor is often asymptomatic, although it can present with symptoms of pain and tooth mobility. A characteristic radiographic appearance is that of a triangular-shaped or semi-circular lucency associated with the roots of erupted teeth. Histologically, the tumor is characterized by the formation of variably sized nests and cords of uniform, benign-appearing, squamous epithelium with occasional vacuolization and keratinization. Treatment of SOT by conservative surgical excision is normally curative with rare episodes of recurrence reported. Since the clinical presentation of SOT may mimic more common pathologic entities, this case report reinforces the need for careful histologic evaluation of all lesions found in the periodontium. PMID- 12083540 TI - Oral pemphigoid masquerading as necrotizing ulcerative gingivitis in a child. AB - BACKGROUND: Cicatricial pemphigoid (benign mucous membrane pemphigoid) is an autoimmune vesiculobullous disease that affects mucosal tissues of adults and rarely presents in children. Only 9 cases in the English literature have reported cicatricial pemphigoid in children, primarily as oral mucosal lesions. This paper presents a case of childhood cicatricial pemphigoid that clinically manifested as necrotizing ulcerative gingivitis (NUG). METHODS: A 9-year-old girl presented with gingival bleeding and discomfort for 2 weeks. NUG was suspected and the patient was treated with antibiotics and an oral hygiene regimen. When the condition did not improve after repeated treatment trials, routine hematoxylin and eosin (H&E) and direct immunofluorescence examinations were performed. RESULTS: Microscopic examination of H&E stained sections showed a non-specific ulceration with chronic inflammation. Direct immunofluorescence studies of peri lesional tissue showed linear deposition of C3 at the basement membrane zone that was consistent with a diagnosis of cicatricial pemphigoid. CONCLUSION: Cicatricial pemphigoid is an autoimmune ulcerative condition that is rarely seen in children. Immunofluorescence studies are essential to differentiate this condition from other ulcerative oral lesions. PMID- 12083541 TI - Oral manifestations of acute myelomonocytic leukemia: a case report and review of the classification of leukemias. AB - BACKGROUND: Oral signs and symptoms may indicate a serious underlying systemic disease. The most frequently observed oral findings of leukemia are mucosal bleeding and ulceration, petechiae, and gingival hyperplasia. This case report describes a 53-year-old male who presented with gingival enlargement and bleeding, fatigue, and recent weight loss as initial manifestations of acute myelomonocytic leukemia. METHODS: A gingival biopsy was performed, revealing the presence of a hypercellular infiltrate of atypical myeloid and monocytic cells. Further work-up consisted of a complete blood count, bone marrow biopsy, and immunohistochemical and histochemical analysis of biopsy material and flow cytometry of peripheral blood. RESULTS: Flow cytometry results confirmed that the infiltrate was of a myelomonocytic origin, and a diagnosis of acute myelomonocytic leukemia was rendered. The patient responded well to a chemotherapeutic induction regimen of cytosine arabinoside and idarubicin hydrochloride, with regression of gingival enlargement and remission of disease. The patient continued with consolidation chemotherapy and an autologous bone marrow transplant, but eventually died 22 months after initial diagnosis. CONCLUSIONS: Oral health care professionals, especially periodontists, must recognize that gingival enlargement may represent an initial manifestation of an underlying systemic disease. Acute myelogenous leukemia is a hematological disorder with a predilection for gingival involvement. PMID- 12083542 TI - Implant placement in maxillary first premolar fresh extraction sockets: description of technique and report of preliminary results. AB - BACKGROUND: The postextraction morphology of the maxillary first premolar extraction socket presents a number of challenges to clinicians seeking ideal implant position, including the morphology of the lateral walls of the extraction socket and the presence of the interradicular septum. A technique for simplification of implant placement at the time of maxillary first premolar extraction is described. METHODS: Sixty-three implants were placed in maxillary first premolar immediate extraction sockets in 57 patients (36 females and 21 males), utilizing a technique which includes removal of residual interradicular bone prior to preparing the osteotomy, use of the removed interradicular bone in the extraction socket defect surrounding the implant, and swaging of the buccal and palatal osseous plates against the implant. RESULTS: All implants demonstrated clinical stability upon uncovering. Forty-one of the implants placed have been restored and in function for a period of up to 2 years. CONCLUSIONS: The technique described affords a simplified and predictable manner for placement of implants into immediate maxillary first premolar extraction sockets. Further studies should be carried out to document long-term success and failure rates of implants placed utilizing this technique, and subsequently restored. PMID- 12083543 TI - Aggressive periodontitis: 5-year follow-up of treatment. AB - BACKGROUND: The hypothesis that in subjects with aggressive periodontitis, a long term stability of periodontal health can be achieved following comprehensive mechanical/surgical and systemic antimicrobial therapy was tested in this prospective study. METHODS: Thirteen patients (36.9+/-7.4 years) with aggressive periodontitis were monitored before and up to 5 years following periodontal therapy. Clinical attachment levels (CAL) were assessed pretherapy, and at 3 months following completion of active periodontal therapy supplemented by amoxicillin plus metronidazole. All subjects were subsequently enrolled in a maintenance program and provided with supportive periodontal therapy with 3 to 4 appointments annually. Reexaminations were performed after 6 months and 1, 2, 3, 4, and 5 years. The data were analyzed using the method of generalized estimating equations (GEE) for CAL changes from baseline to the 3-month visit, and from completion of periodontal therapy to each annual visit up to the 5-year follow-up reappointment. RESULTS: During the 5-year study, all subjects strongly benefited from periodontal treatment. Between baseline and the 3-month reexamination, the CAL levels revealed a significant decrease of 2.23 mm (95% confidence interval [CI]: 1.77 to 2.69 mm; P < or =0.001). At the 5-year maintenance visit, the CAL changes ranged from -0.04 to +0.29 mm with no further statistically significant periodontal breakdown (P >0.05). Five years after surgery, 3.2% of the treated sites demonstrated a further CAL gain > or =3 mm. A stabilization (CAL -2 to +2 mm) occurred in 94.6% of the cases. The number of periodontal sites experiencing a breakdown varied from 5.3% at 6 months to 2.2% at 5 years. CONCLUSIONS: In aggressive periodontitis, comprehensive mechanical/surgical and antimicrobial therapy is an appropriate treatment regimen for long-term stabilization of periodontal health. In this study, periodontal disease progression was successfully arrested in 95% of the initially compromised lesions, while 2% to 5% experienced discrete or recurrent episodes of loss of periodontal support. PMID- 12083544 TI - Bacteria-modified amperometric immunosensor for a Brucella melitensis antibody assay. AB - A novel amperometric immunosensor setup is described which uses horseradish peroxidase (HRP) as a label in conjunction with a current-based Brucella sensor. The Bacteria modified immunosensor was constructed by using a biocomposite formed by dispersing graphite powder into a mixture of Brucella melitensis and silicate polymer gel. The enzyme-labeled antibody can readily diffuse toward the encapsulated antigen (Brucella melitensis), which retains its binding properties, and the association reaction is easily detected at the surface exposed to the solution. The use of an oaminophenol (o-AP) substrate and amperometric detection at -150 mV (vs. SCE) results in a relatively low detection limit of 3.5 ng/ml and a linear detection range of 3.5 ng/ml to 200 ng/ml. Based on an optimized parameter, the prepared sensor was used to detect the Brucella melitensis antibody in serum samples by using a competitive binding assay. The results demonstrate the feasibility of employing the proposed immunosensor for the detection for Brucella melitensis antibody in a clinical analysis. PMID- 12083545 TI - A simple HPLC-fluorescence detection of nitric oxide in cultivated plant cells by in situ derivatization with 2,3-diaminonaphthalene. AB - An HPLC method with fluorescence detection for the determination of nitric oxide (NO) in cultivated plant cells (Agave pacifica, Agavaceae) was developed. NO was derivatized in situ with 2,3-diaminonaphthalene (DAN) as a labeling reagent and converted to 1(H)-naphthotriazole. The maximum peak height of the derivative was observed by incubation for 3 h at 25 degrees C with 0.2 mM DAN. Excess reagent in cells was removed by washing 3 times with 5 ml of water. The calibration curve for authentic standard of DAN-NO spiked to cultivated plant cells showed a good linearity (r = 0.995) in the range of 5.0 to 50 pmol/g cell. The detection limit at a signal-to-noise ratio of 3 was 3.4 pmol/g cells. The proposed method was successfully applied to the monitoring of NO concentration with cell growth. The effect of thermal treatment on the concentration of NO in plant cells was also examined. The concentration of NO in cells treated at 5 degrees C for 1 h was significantly higher than that treated at 25 degrees C and 35 degrees C for 1 h (n = 3, p < 0.05). PMID- 12083547 TI - Deviations of capacitive and inductive loops in the electrochemical impedance of a dissolving iron electrode. AB - The electrochemical impedance of an iron electrode often shows the capacitive and inductive loops on the complex plane. The capacitive loop originates from the time constant of the charge transfer resistance and the electric double layer capacitance. The inductive loop is explained by Faradaic processes involving the reaction intermediate. In some cases, these loops deviate from a true semicircle. In this paper, the origins and curve-fitting methods for the deviated loops of electrochemical impedance are discussed. The constant phase element (CPE) was used to present the deviation of the capacitive loop instead of electric double layer capacitance. The reaction rate constants, which are a function of the frequency, are proposed for the Faradaic impedance to present the deviated inductive loop. PMID- 12083546 TI - Electrocatalytic response of dopamine at a metallothioneins self-assembled gold electrode. AB - The metallothioneins (MT) self-assembled monolayer modified gold electrode (MT/Au) is demonstrated to catalyze the electrochemical response of dopamine (DA) by cyclic voltammetry. A pair of well-defined redox waves was obtained and the calculated standard rate constant (k(s)) is 6.97 x 10(-3) cm s(-1) (20 degrees C) at the self-assembled electrode. The electrode reaction is a quasi-reversible process. The oxidation peak of DA can be used to determine the concentration of DA. The peak current and the concentration of DA follow a linear relationship in the range of 2.0 x 10(-5) M to 8.0 x 10(-4) M. The detection limit is 6.0 x 10-6 M. By ac impedance spectroscopy, the apparent electron transfer rate constant (k(app)) of Fe(CN)6(3-)/Fe(CN)6(4-) at the MT/Au electrode was obtained as 2.0 x 10(-5) cm s(-1). The MT/Au was characterized with grazing angle FT-IR spectroscopy and contact angle goniometry. PMID- 12083548 TI - Spectroelectrochemistry study on the electrochemical reduction of ethidium bromide. AB - The electrochemical reduction mechanism of ethidium bromide was first studied by spectroelectrochemistry. This reduction was proved to be a two-step process by cyclic voltammetry, differential pulse voltammetry and spectroelectrochemistry, in which each step was proved to be a one-electron transfer process by a spectropotentiostatic fluorescence technique. Hydroethidine was confirmed to be the final product by comparing the spectrum of the product of the electrochemical reduction to that of the product of the chemical reduction of ethidium bromide, and a carbon-centered radical was concluded to be a reasonable intermediate product during the electrochemical reduction of ethidium bromide. PMID- 12083549 TI - Speciation of molybdenum in river water by size fractionation and catalytic determination. AB - A speciation scheme of trace molybdenum was proposed for river water based on size fractionation by filtration and ultrafiltration and the catalytic spectrophotometric determination of the reactive molybdenum concentration (CR). The total concentration (CT) of molybdenum was determined by the same method after acid decomposition to obtain the concentration (CT - CR) of unreactive molybdenum. Most molybdenum in natural river-water samples was found to be reactive species. A large part of the molybdenum was found in the fraction of molecular weight (MW) < 10(3), and was estimated to be MoO4(2-) from the chemical equilibria of molybdate ions. The residual part of molybdenum was found in the colloidal and particle fractions (MW > or = 10(4)), and was characterized as reactive molybdenum adsorbed or complexed on humic iron aggregates. The coexistence of silicate contributed to a decrease of the particle size of humic iron aggregates associated with molybdenum. The above-mentioned speciation results were confirmed by an analysis of artificial samples. The changes in the fractionation results by acidification (0.1 M HCl) were also used to characterize molybdenum in natural water. PMID- 12083550 TI - Application of a pentafluorobenzyl bromide derivatization method in gas chromatography/mass spectrometry of trace levels of halogenated phenols in air, water and sediment samples. AB - An analytical method using pentafluorobenzyl bromide (PFBB) derivatization and gas chromatography/mass spectrometry (GC/MS) has been applied to identify and quantify chloro-, bromo- and dichlorophenols in air, water and sediment samples. Phenols in air sample were collected with a PS-2 Sep-PAK cartridge, and eluted with 2-propanol. For water and sediment samples, liquid-liquid extraction with dichloromethane was carried out, and the solvent was exchanged to 2-propanol. The phenols in the solution reacted with PFBB to form the corresponding pentafluorobenzyl esters. After extracting the derivatives into hexane, the determination was carried out by GC/MS with selected-ion monitoring. The detection limits of phenols in air, water and sediment were 0.0033 - 0.0073 microg/m3, 0.0066 - 0.0147 microg/L and 0.33 - 0.73 microg/kg, respectively. More than 90% recoveries of the halogenated phenols were obtained from real environmental samples spiked by the halogenated phenols. The three isomers of mono-chlorophenols were detected in sediment samples in the range of 5.2 - 9.2 microg/kg in wet weight basis. PMID- 12083551 TI - Optimization of high-performance liquid chromatographic parameters for the determination of capsaicinoid compounds using the simplex method. AB - A high-performance liquid chromatographic method was developed for the analysis of capsaicinoid compounds, the pungent principles of capsicum fruits. A sequential simplex method was applied to optimize the chromatographic response function used to assess the quality of separation by varying the chromatographic parameters. The separation was achieved in 11 min using a C-8 column of 15-cm length and 4.6 mm diameter using a UV detector. A flow rate of 1.15 ml min(-1) at a column temperature of 43.5 degrees C using 63.7% methanol in water gave the most efficient separation. The method was found to be suitable for the determination of the major capsaicinoid compounds in the capsicum samples. PMID- 12083552 TI - Sensitive kinetic-spectrophotometric determination of iodate in iodized table salt based on its accelerating effect on the reaction of bromate with chloride ion in the presence of hydrazine. AB - A simple, precise, sensitive and accurate method was developed for rapid determination of trace quantities of iodate. The method is based on the accelerating effect of iodate on the reaction of bromate and chloride acid in the presence of hydrazine in acidic media. The decolorization of Methyl Orange with the reaction products was used to monitor the reaction spectrophotometrically at 525 nm. Iodate could be determined in the concentration ranges of 0.03 - 1.2 microg ml(-1). The relative standard deviation for ten replicate determinations of 0.3 microg ml(-1) of iodate was 1.65%. The proposed method was applied to the determination of iodate in table salts with satisfactory results. PMID- 12083553 TI - Indirect spectrophotometric determination of propranolol hydrochloride and piroxicam in pure and pharmaceutical formulations. AB - Two simple and sensitive indirect spectrophotometric methods for the assay of propranolol hydrochloride (PPH) and piroxicam (PX) in pure and pharmaceutical formulations have been proposed. The methods are based on the oxidation of PPH by a known excess of standard N-bromosuccinimide (NBS) and PX by ceric ammonium sulfate (CAS) in an acidic medium followed by the reaction of excess oxidant with promethazine hydrochloride (PMH) and methdilazine hydrochloride (MDH) to yield red-colored products. The absorbance values decreased linearly with increasing concentration of the drugs. The systems obeyed Beer's law over the concentration ranges of 0.5 - 12.5 and 0.3 - 16.0 microg/ml for PPH, and 0.4 - 7.5 and 0.2 - 10 microg/ml for PX with PMH and MDH, respectively. Molar absorptivity values, as calculated from Beer's law data, were found to be 1.36 x 10(4) and 2.55 x 10(4) l mol(-1) cm(-1) for PPH, and 2.08 x 10(4) and 2.05 x 10(4) l mol(-1) cm(-1) for PX with PMH and MDH, respectively. The common excipients and additives did not interfere with their determinations. The proposed methods have been successfully applied to the determinations of PPH and PX in various dosage forms. The results obtained by the proposed methods compare favorably with those of official methods. PMID- 12083554 TI - A new strategy for exploiting ion exchange in sequential injection analysis: in line phytic acid separation/determination in foods as an example. AB - A novel strategy for exploiting ion exchange in sequential injection systems is proposed. The procedure is based on the selection of a defined volume of a resin suspension, which is introduced and packed in the analytical path, establishing a resin mini-column in the system. The passage of a selected sample volume through the resin mini-column leads to the retention of the analyte, while the sample matrix is discarded. The analyte is eluted during the passage of the eluant/reagent by the packed beads, being the analytical signal monitored (absorbance) in the liquid phase. The beads are then aspirated back to the holding coil and directed to a recovery flask, linked at the selection valve; then the system is ready to begin a new cycle. With the proposed strategy, the main characteristics of the sequential injection system are kept as any new artifact is added to the manifold and system reconfiguration is not required. The feasibility of the approach is demonstrated by the phytic acid determination in food samples. For this specific application, AG1-X8 was selected as ion exchanger, and a solution containing Cl- and Fe(III)-salicylate complex was used as eluant and spectrophotometric reagent. PMID- 12083555 TI - Behavior of bound water in polyethylene oxide studied by DSC and high-frequency spectroscopy. AB - Measurements of DSC, NMR and high-frequency spectroscopy have been done on PEO H2O and PEO-H2O-HQ mixed systems. DSC measurements show that water molecules contained in PEO are divided roughly into two species: freezable water and non freezable water. By 1H-NMR measurements, PEO and H2O protons show an absorption peak around 3.6 ppm and 4.5 ppm, respectively. All the PEO-H2O samples containing H2O show NMR absorption signals, suggesting the existence of movable H2O. Samples with H2O molar ratio over 0.5, show one absorption peak of H2O protons, and more than two peaks of PEO. The results from DSC and NMR measurements show that the bound state of H2O in PEO, as well as the mobility of PEO itself, varies according to the water content in PEO-H2O samples. On the other hand, DSC measurements give the result that PEO-H2O-HQ systems can be considered as homogeneous so long as the content of H2O and/or HQ is not so high. Furthermore, the water molecules in the system exist as bound water. Results of high-frequency spectroscopic measurements for the PEO-H2O and PEO-H2O-HQ systems show good agreement with those of DSC and NMR measurements. PMID- 12083556 TI - Simultaneous determination of endocrine disruptive alkylphenolic compounds as dansyl derivatives. PMID- 12083557 TI - Fluorescence dynamic anisotropy of spinach calmodulin labeled by a fluorescein chromophore at Cys-26. PMID- 12083558 TI - Flow injection chemiluminescence determination of amino acids by oxidation with N bromosuccinimide. PMID- 12083559 TI - Micro-phase sorbent extraction for trace analysis via in situ sorbent formation: application to the preconcentration and the spectrophotometric determination of trace ammonia. PMID- 12083560 TI - Application of an orthogonal method for optimizing an inductively coupled plasma mass spectrometer. AB - The orthogonal method can be used to predict the optimum instrumental conditions and to guide the optimizing operations. This simplifies the ICP-MS optimization. In general, the final optimum parameter combination used in an analysis can be obtained after fine adjustments of the preferred parameters derived from the orthogonal experiment. This method can also be used to study the relationship between the parameter settings and the signal intensity deflection to improve the analysis of elements of a limited mass range or of isotopes. PMID- 12083561 TI - Evaluation of several gas chromatographic stationary phases for the solid-phase extraction of trace amounts of copper in aqueous samples. PMID- 12083562 TI - Application of capillary electrophoresis with sensitive detection to analysis for saccharide molecules. PMID- 12083563 TI - Selective fluorescent molecular sensing based on 2A,2D-disulfonyl dibenzosulfolane-diphenyl-capped beta-cyclodextrin for phenolic guests. PMID- 12083564 TI - In situ UV-VIS detection of hydrothermal reactions using fused-silica capillary tubing within 0.08-3.2 s at high temperatures. PMID- 12083565 TI - Comparative study of a cluster analysis and a principal-component analysis using a polarized imaging technique for discriminating adhesive cloth tapes. PMID- 12083566 TI - Crystal structure of 3-amino-1,2,4-triazin-5(2H)-one. PMID- 12083567 TI - Crystal structure of 4-(1-mesityl-1-methylcyclobutane-3-yl)-2-(N ethyl)aminothiazole. PMID- 12083568 TI - Crystal structure of [[mu-N,N'-bis(salicylidene)-1,3 propanediaminatodimethylformamide]zinc(II)]diiodozinc(II). PMID- 12083569 TI - Crystal structure of (-)-(S)-N-(endo-8-methyl-8-azabicyclo[3.2.1]oct-3alpha-yl)-4 amino-5-chloro-2-(1-methyl-2-butynyl)oxybenzamide monohydrochloride tetrahemihydrate. PMID- 12083570 TI - Minimizing operative time: does speed kill? PMID- 12083571 TI - Pain in the thrower's shoulder: how much should little leaguers be pitching? PMID- 12083572 TI - Radiologic case study. Bizarre parosteal osteochondromatous proliferation (Nora's lesion) of bone. PMID- 12083573 TI - Surgical templates for orthopedic operative reports. AB - Generating an operative report after a surgical procedure is a critical but time consuming and expensive task for every physician. Use of a template can shorten this process. This article describes an easy method by which a physician can create a template that can be customized to allow discussion of unique features of a given case. PMID- 12083574 TI - Percutaneous compression plating for intertrochanteric hip fractures: treatment rationale. AB - Percutaneous compression plating was developed after thorough consideration of each stage in the surgical procedure for intertrochanteric fractures and the ways in which they might be improved. This system represents a minimally invasive method of fracture stabilization and fixation, providing enhanced rotational stability and bone sparing, which reduces both peri- and postoperative complications including cut outs and fracture collapse. PMID- 12083575 TI - Spondylolysis: returning the athlete to sports participation with brace treatment. AB - Between 1988 and 1995, 73 adolescent athletes treated with the Boston Overlap Brace for spondylolysis were reviewed to evaluate improvement in pain score and activity level. A favorable clinical outcome was achieved in 80%. Girls and boys who participated in high-risk sports were five times more likely to have an unfavorable clinical outcome than those who participated in low-risk sports (odds ratio = 5, 95% confidence interval = 2.4-7.5, P = .003). In addition, acute onset of pain and hamstring tightness were associated with a worse outcome. Athletes with symptomatic spondylolysis treated with an antilordotic brace can expect improvement in their clinical course and return to sports participation in 4-6 weeks. PMID- 12083576 TI - Biomechanical testing of bioabsorbable cannulated screws for slipped capital femoral epiphysis fixation. AB - This study compared cannulated 4.5-mm bioabsorbable screws made of self reinforced poly-levolactic acid to cannulated 4.5-mm steel and titanium screws for resistance to shear stress and ability to generate compression in a polyurethane foam model of slipped capital femoral epiphysis fixation. The maximum shear stress resisted by the three screw types was similar (self reinforced poly-levolactic acid 371 +/- 146 MPa, steel 442 +/- 43 MPa, and titanium 470 +/- 91 MPa). The maximum compression generated by both the self reinforced poly-levolactic acid screw (68.5 +/- 3.3 N) and the steel screw (63.3 +/- 5.9 N) was greater than that for the titanium screw (3 +/- 1.4 N, P <.05). These data suggest cannulated self-reinforced poly-levolactic acid screws can be used in the treatment of slipped capital femoral epiphysis because of their sufficient biomechanical strength. PMID- 12083577 TI - Elbow heterotopic ossification in head-trauma patients: diagnosis and treatment. AB - Heterotopic ossification is a disorder characterized histologically and radiographically by normal bone formation in soft tissues that normally have no ossification properties. In severe head-trauma patients, a high incidence of heterotopic ossification occurs. The diagnosis of this pathology in these patients often is difficult for residual neurological damage. The etiology of heterotopic ossification following head trauma is unknown. Similarities have been found between heterotopic ossification and myositis ossificans, a hereditary autosomal dominant disease. PMID- 12083578 TI - The relationship between serum relaxin and knee joint laxity in female athletes. AB - Gravidity studies have intimated a relationship between serum relaxin levels and joint laxity. To assess its applicability to eumenorrheic athletes, weekly serum relaxin and knee arthrometry measurements were obtained in 57 collegiate women and 5 men for 4 consecutive weeks. The former included athletes, ACL-injured athletes, and nonathletes, while the latter formed an a priori hormonal control. Analysis of variance revealed a significant change in weekly serum relaxin levels in females but not in anterior translation. Regression analysis failed to disclose a relationship between these variables. Therefore, we conclude relaxin does not affect knee laxity. PMID- 12083579 TI - Synovial abrasion for isolated, partial thickness, undersurface, medial meniscus tears. AB - Synovial abrasion and rasping provides good healing potential of isolated, partial thickness, undersurface medial meniscal tears. It is an effective, easy, and valuable treatment modality. PMID- 12083580 TI - Isolated necrosis of the brachialis muscle due to exercise. PMID- 12083581 TI - Simultaneous tears of the Achilles tendon and medial head of the gastrocnemius muscle. PMID- 12083583 TI - The resident as a working stiff. PMID- 12083582 TI - Scurvy presenting as a medial head tear of the gastrocnemius. PMID- 12083584 TI - Thalassemia major. PMID- 12083585 TI - Managing disorders of the cervicothoracic junction. AB - A wide range of pathologic conditions occur at the cervicothoracic junction. These conditions are usually the result of trauma, neoplastic processes, infection, prior surgery, or degenerative changes. Instability in this region of the spine is difficult to manage, particularly because of the complex biomechanics involved and the challenging surgical approaches required for treatment. Traditional radiologic evaluation of the cervicothoracic junction is often inadequate; as a result, the standard 3-view cervical spine series should be augmented with swimmer's or oblique views. Surgical treatment, designed to increase stability and allow early mobilization and rehabilitation, often requires internal fixation devices; lateral-mass or pedicle screws are increasingly being used to avoid complications associated with devices occupying the spinal canal. Although posterior surgical approaches to the cervicothoracic junction are relatively straightforward, anterior approaches require mastery in traversing the various bony and soft-tissue structures. PMID- 12083586 TI - Magnetic resonance imaging analysis of coracoid morphology and its relation to rotator cuff tears. AB - The purpose of this study was to define the magnetic resonance imaging (MRI) anatomy of the subcoracoid space and to determine its relation to rotator cuff tears. Routine clinical MRI sequences of 100 shoulders were reviewed, and standardized measurements of the subcoracoid space were taken and compared with previously published computed tomographic (CT) data. MRI and CT values were very similar. There were no significant differences between the coracoid morphology of patients with normal findings and patients with varying degrees of rotator cuff disease involving the supraspinatus tendon. The excellent agreement between MRI and CT measurements indicates that different investigators may use either imaging modality to obtain reproducible results. The role of coracoid anatomy in the development of lesions of the subscapularis tendon and the long head of the biceps remains to be defined. PMID- 12083587 TI - Intravenous parecoxib sodium foracute pain after orthopedic knee surgery. AB - Our objective in a randomized, multicenter, double-blind, parallel-group, placebo and active-controlled study was to evaluate and compare the analgesic effectiveness of single intravenous (IV) doses of parecoxib sodium 20 and 40 mg, morphine 4 mg, and ketorolac 30 mg in the postsurgical orthopedic pain model. After undergoing unilateral total knee replacement surgery, 208 healthy adult patients were randomized to receive placebo or a study drug within 6 hours of discontinuation of patient-controlled analgesia on postoperative day 1. Onset of analgesia was similarly rapid with IV parecoxib sodium 40 mg, morphine, and ketorolac. Level and duration of analgesia were significantly superior with parecoxib sodium than with morphine and were similar for parecoxib sodium and ketorolac. Parecoxib sodium was safe and well tolerated. In conclusion, IV parecoxib sodium 40 mg is as effective as ketorolac 30 mg and is more effective than morphine 4 mg and therefore has potential widespread utility in acute postoperative pain management. PMID- 12083588 TI - Atypical presentation of an osteoid osteoma in a child. AB - Osteoid osteomas are benign bone tumors that most commonly involve the long bones of the lower extremities, but do occur in the spine with some frequency. Patients with lesions in the spine typically present with back pain, scoliosis, and, less commonly, varying degrees of radicular leg pain. We report the case of a child with complaints of nonradicular leg pain. A combination of plain radiographs, scintigraphy, and axial imaging showed a lesion in the sacral spine consistent with an osteoid osteoma. After failed medical management, our patient was treated with intralesional excision, with complete resolution of symptoms. PMID- 12083589 TI - Destructive posttraumatic subtalar synovitis. AB - Destructive posttraumatic synovitis is a rare entity usually involving the knee. This case report details the evaluation and treatment of a rapidly progressive and destructive subtalar lesion that occurred in a healthy 27-year-old man after incidental trauma. Initial orthopedic evaluation revealed diffuse ankle tenderness and mild effusion. Radiographic and laboratory studies showed normal findings. Bone scan revealed significantly increased uptake in the talus. Subsequent computed tomography scan and magnetic resonance imaging studies showed a lateral soft-tissue mass invading both the talus and calcaneus with destruction of the subtalar joint. Tissue biopsy samples revealed hypertrophic synovium with villous projections, occasional giant cells, and occasional hemosiderin deposits. The lesion caused rapid and progressive destruction of the subtalar joint despite aggressive conservative measures. Ultimately, the patient underwent a subtalar arthrodesis that led to the resolution of his symptoms and resumption of work and recreational activities. While posttraumatic synovitis is a rarely encountered condition, the orthopedic surgeon must have an index of suspicion, perform a systematic evaluation, and render definitive treatment to ensure a successful outcome. PMID- 12083590 TI - Breakage of a 1.3-mm AO/ASIF titanium plate after phalangeal osteotomy in two patients. AB - Two patients sustained plate breakage after a proximal phalanx corrective osteotomy stabilized with an AO/ASIF titanium 1.3-mm plate. The failure of this plate after the initiation of early range of motion therapy warrants further evaluation regarding its inherent strength and clinical application. PMID- 12083591 TI - The thrower's shoulder. Part II: a surgical discussion. PMID- 12083592 TI - Publication rates of presentations made at annual meetings of the American Orthopaedic Society for Sports Medicine and the Arthroscopy Association of North America. AB - The purpose of this study was to determine the publication rates of presentations made at the annual meetings of 2 sports medicine specialty societies--the American Orthopaedic Society for Sports Medicine (AOSSM) and the Arthroscopy Association of North America (AANA). We created a database covering annual AOSSM meetings from 1990 to 1993 (4 years) and annual AANA meetings from 1991 to 1993 (3 years) and searched the Melvyl Medline Plus database for abstracts from 1990 through 1998 to determine which had been published in peer-reviewed journals. Of the 333 abstracts listed for the 1990 to 1993 meetings, 198 (59.5%) were published in peer-reviewed journals. Publication rates of the AOSSM and AANA meetings were 68.1% and 50.9%, respectively. The majority of articles were published in American Journal of Sports Medicine (40.1%) and Arthroscopy (30.3%). Publication rates of presentations made at meetings of these sports medicine specialty societies are high and exceed the publication rates associated with meetings of other medicine specialty societies. PMID- 12083593 TI - Emergent psychopathology in Alzheimer's disease patients over 12 months associated with functional, not cognitive, changes. AB - In a large, well-characterized population of community-dwelling persons with Alzheimer's disease (AD), we investigated the emergence of behavioral symptomatology and its association with changes in cognitive, global-clinical, and functional status. Behavioral Rating Scale for Dementia (BRSD) item responses from 235 AD patients with varying levels of dementia severity and without significant behavioral disturbance were taken from the baseline and 12-month visits in a study of cognitive and behavioral instruments. Item-level analysis revealed new symptoms at every dementia severity level. The symptoms that emerged in the greatest proportion of patients were change in weight, change in appetite, diurnal confusion, uncooperativeness, restlessness, clingy behavior, loss of initiative, and change in sleeping pattern. Changes in cognitive status over the 12 months were associated with changes in functional status and not with the emergence of behavioral symptomatology; however, change in the latter two domains tended to be associated. The findings support the hypothesis that increasing behavioral disturbance is not strongly associated with decreasing cognitive status and that, except for psychotic symptoms, a previously observed association between dementia severity and behavioral status may have been mediated partly by changes in functional abilities. PMID- 12083594 TI - Greater daytime sleepiness in subcortical stroke relative to Parkinson's disease and Alzheimer's disease. AB - Deficits in daytime alertness in the elderly may reflect, in part, deterioration of the critical neural systems modulating circadian control of sleep and wakefulness. In this study, 47 patients with subcortical stroke (n = 9), Alzheimer's disease (n = 6), and parkinsonism (n = 32) underwent a 24-hour in laboratory evaluation consisting of overnight polysomnography and next-day evaluation of daytime sleep tendency with the Multiple Sleep Latency Test. Patients with stroke were significantly sleepier during the daytime relative to the Parkinson's and Alzheimer's disease patients. Nocturnal sleep parameters did not account for these differences. In the stroke patients, some infarcts occurred in the vasculature, impacting blood supply to the hypothalamus. We interpret these effects as representing functional interruptions of the suprachiasmatic nucleus (SCN). These results are also compatible with a wake-promoting function of the human SCN. PMID- 12083595 TI - Relationship between Alzheimer's disease severity and patient participation in decisions about their medical care. AB - To examine the relationships between Alzheimer's disease (AD) severity and patient participation in decisions about their medical care, we performed a cross sectional study of 74 caregivers' ratings of the degree to which AD patients participate in making decisions about their medical care. The majority of the caregivers reported that they made the final decisions about medical care (n = 48, 64%), one-quarter (n = 19, 26%) reported an equal role, and a small proportion (n = 7, 9%) reported that the patient made the final decisions. Multivariate models showed that the significant predictors of the patient's participation in decisions about medical care were measures of dementia severity: Mini-Mental State Examination (MMSE) and Clinical Dementia Rating (CDR). Patients with mild-stage disease (CDR = 1 or MMSE > or = 20) are involved in decision making largely in a collaborative role with the caregiver. Beyond mild-stage disease, the clinician can largely expect that the caregiver is making the final decisions. These results illustrate how and when caregivers can expect changes in their decision making role. PMID- 12083596 TI - Clinical outcome and adverse effects of electroconvulsive therapy in elderly psychiatric patients. AB - We evaluated the short-term outcome and side effects of electroconvulsive treatment (ECT) in a population of elderly psychiatric patients. The material included patients consecutively hospitalized at a psychogeriatric department within a period of 5 years. From a total number of 239 patients, we included 52 treated with ECT (22%). Altogether, 41 patients (79%) improved clinically. Of them, 21 (40%) of the patients went home after the treatment. Twenty patients (38%) developed adverse reactions from the ECT treatment. Impaired memory (14%), confusion (6%), and hypertension (6%) represented the most commonly reported negative reactions secondary to the treatment. The findings from the study support previous assumptions that ECT is effective in the treatment of serious psychiatric disorders in elderly patients. Because of the relative high frequency of side effects, the patients should be selected PMID- 12083597 TI - Risperidone and 9-hydroxyrisperidone concentrations are not dependent on age or creatinine clearance among elderly subjects. AB - Risperidone is extensively metabolized to an active metabolite, 9 hydroxyrisperidone (9-OH), which is dependent on renal clearance. Risperidone and 9-OH clearances are reduced in the elderly when compared to young subjects. The objective of this study was to determine whether among elderly subjects, risperidone and 9-OH clearance would further decline with increasing age and decreasing creatinine clearance (CrCl). Twenty geriatric inpatients were evaluated in a naturalistic setting with regard to total daily risperidone dose and dosing interval. Creatinine clearance was determined using an 8-hour urine collection. Risperidone and 9-OH concentrations were determined by radioimmunoassay. Spearman's correlation coefficients were used to examine the impact of age and CrCl on concentrations of risperidone, 9-OH, their sum, and the quotient of 9-OH/risperidone. Mean age was 76.4 +/- 9 years (range 56-91). Mean CrCl was 55.4 +/- 32.8 mL/min/1.73 m2 (range 17-142 mL/min/1.73 m2). Mean risperidone daily dose was 1.3 +/- 0.7 mg. Steady-state risperidone and 9-OH concentrations were 4.1 +/- 5.3 ng/mL and 9.1 +/- 6.2 ng/mL, respectively. Mean 9 OH/risperidone was 6.2 +/- 6.1. Concentrations of risperidone, 9-OH, their sum, and 9-OH/risperidone were not significantly correlated with age or CrCl. These results were unchanged when concentrations were corrected for total daily risperidone dose. Among elderly subjects, risperidone and 9-OH clearance do not decline with increasing age or declining CrCl. PMID- 12083598 TI - Hippocampal atrophy disrupts transfer generalization in nondemented elderly. AB - Specific reductions in hippocampal volume in nondemented elderly individuals with mild cognitive impairment have been shown to correlate with future development of Alzheimer's disease (AD). Hippocampal atrophy (HA) is also correlated with cognitive impairments, leading to the promise of behavioral markers for early AD. Prior theoretical work has suggested that hippocampal dysfunction may selectively impair generalization involving novel recombinations of familiar stimuli. In this study, nondemented elderly individuals were trained on a series of concurrent visual discriminations and were then tested for transfer when stimulus features were recombined in new ways. Presence or absence of HA, revealed by neuroimaging, was not correlated with concurrent discrimination performance; however, individuals with mild HA showed significant decreases in transfer performance relative to nonatrophied participants. These preliminary results suggest that even very mild degrees of hippocampal atrophy may be associated with subtle behavioral impairments. PMID- 12083599 TI - Change in the rate of restraint use and falls on a psychogeriatric inpatient unit: impact of the health care financing administration's new restraint and seclusion standards for hospitals. AB - This chart review of 767 patients treated on a psychogeriatric inpatient unit over a 2-year period examined the impact of the Health Care Financing Administration's (HCFA) new restraint standards for hospitals on the rate of restraint use and falls. There was a marked decrease in the number of restraint episodes in the year after the introduction of the new standards compared with the year before (44 vs 212 restraint episodes per 1,000 patient-days). However, no notable differences were found in the number of total falls (18 vs 21 falls per 1,000 patient-days) or serious falls (2 vs 1 serious fall per 1,000 patient days). If replicated, these findings of decreased restraint use without a concomitant increase in the number of falls demonstrate a change in practice patterns as a result of HCFA's action. PMID- 12083600 TI - No long-term effect of behavioral treatment on psychotropic drug use for agitation in Alzheimer's disease patients. AB - Th determine if teaching caregivers behavior management techniques (BMTs) reduces long-term psychotropic use in Alzheimer's disease (AD) patients, we examined 12 month follow-up data from a 4-month randomized study comparing placebo, BMTs, trazodone, and haloperidol for the treatment of agitated behaviors in persons with AD. After 4 months, treatment was allowed with any agent. Between 42.8% and 51% of AD patients received additional psychotropics between 4 and 12 months. The relative risk of being prescribed any psychotropic drug after the 4-month trial was at or about 1.0 for subjects in each drug arm or placebo arm versus BMTs. We concluded that teaching caregivers BMTs did not diminish long-term prescription of psychotropic drugs. PMID- 12083602 TI - The Umwelt and its relevance to animal communication: introduction to special issue. AB - This introduction applies J. von Uexkull's (1934/1957) concept of the Umwelt to the study of animal communication, particularly as it pertains to studies presented at a recent workshop on animal communication in the context of the environment. The environment is conceived broadly in the articles that follow, including the many physical and social environments in which an animal may find itself. The Umwelt concept is briefly expanded here to include also the personal microenvironment of the signaler in which the signal is embedded into the suite of concurrent nonsignaling behaviors of the individual. Other animals may even infer aspects of the signaler's own immediate Umwelt by noticing accompanying attentional cues such as the direction of eye gaze. In this way, part of the Umwelt can be accessible to companions, facilitating the communication process. PMID- 12083601 TI - Selective attention skills in differentiating between Alzheimer's disease and normal aging. AB - We determined the reliability and validity of a cancellation test of symbols (Symbol Cancellation Test [SCT]), designed to assess visual selective attention deficits in the elderly, on 34 Alzheimer's disease (AD) patients from Bari University Hospital Center, Bari, Italy, and 232 nondemented elderly subjects, aged 68 to 87 years, from the second prevalence survey (1995 through 1996) of the Italian Longitudinal Study on Aging (Casamassima, Bari, Italy). To assess convergent and discriminant validity, the Digit Cancellation Test (DCT), Mini Mental State Examination (MMSE), and Babcock Story Recall Test (BSRT) were administered. Finally, discriminant accuracy of SCT between AD patients and nondemented elderly subjects was assessed. Inter-rater and test-retest reliability for P1 and P2 was excellent for both AD patients and the normal population, with a high degree of internal consistency. The SCT was significantly correlated with the DCT (0.67), MMSE (0.60), and BSRT (0.33). The classification accuracies of overall performance on the SCT for subjects with and without AD were 0.62 and 0.91, respectively. The SCT is a valid and reliable test to assess selective attention in elderly subjects, in whom dementing illness must be diagnosed and clinically distinct from age-related cognitive decline. PMID- 12083603 TI - Overview: Animal acoustic communication and the role of the physical environment. AB - Scientists from many distinct disciplines often use concepts such as habitat, environment, and niche to describe among other things the physical characteristics of the communicative worlds of animals. Often these descriptions center on the physical characteristics most salient to humans' perceptual systems. In this article, the authors advocate an approach to the description and analysis of the physical characteristics of animal communicative worlds based on J. von Uexkull's (1934/1957) concept of the Umwelt. Such an approach emphasizes the physical characteristics of the world as perceived by the organism in question, and its use has often led to novel and unexpected insights into animal communicative systems. PMID- 12083604 TI - The properties of geophysical fields and their effects on elephants and other animals. AB - Geophysical properties of acoustic, seismic, electric, and magnetic waveforms create opportunities and constraints for animals' communication and sensory monitoring of the environment. The geometric spreading of waves differs; at some frequencies, transmission is most efficient and has minimal noise. The spreading properties of seismic waves favor long-distance propagation for communication and environmental monitoring, and would benefit elephants (Elephas maximus and Loxodonta africana), such as in locating subsurface water. Extending C. E. O'Connell-Rodwell, B. T. Amason, and L. A. Hart (2000), a man jumping at 1.11 km propagated seismic waves at 10-40 Hz. Given the noise of lightning and the Schumann resonances, near field magnetic and electric transmission by animals would be most efficient around 1000 Hz. PMID- 12083605 TI - Auditory communication, meteorology, and the Umwelt. AB - Animal communication is affected by a multitude of environmental factors including meteorological conditions, ground reflection, turbulence, and vegetative attenuation. Evolutionary and behavioral adaptations by animals to these challenges include elephant use of infrasound optimized for long-distance propagation. These adaptations in turn influence the perceptual sphere, or Umwelt, of calling animals. PMID- 12083606 TI - Changes to acoustic communication systems in human-altered environments. AB - Animal communication systems have become closely tuned to local habitat conditions as populations have adjusted to different long-term environmental pressures. However, many habitats are now rapidly changing because of anthropogenic modification. Maintenance of effective communication systems in greatly altered environments will depend on communicative responses on both evolutionary and ontogenetic time scales. Consideration of potential acoustic challenges caused by human-generated habitat modification has important implications for basic research and conservation biology. First, the observed signal structure of individuals in altered environments may not match the normally hypothesized call structure. Second, species that either possess little ability to adapt quickly on an evolutionary time scale or have little plasticity in their communicative systems may be unable to respond to large anthropogenic alterations in their acoustic environment. PMID- 12083607 TI - Overview: Signal production and detection. AB - Studies of signal production and detection are complicated by the diverse modalities used by animals, constraints of human signal perception, and limitations of a controlled laboratory environment. These complications can be addressed by creative approaches to studying anatomical adaptations for signal production and detection, innovative technology, and careful studies of signal production and detection in the field. These solutions should overcome the perceptual limitations of researchers as they explore Umwelten in other species. PMID- 12083608 TI - The production of communication signals at the air-water and water-substrate boundaries. AB - The 2 interfaces of the aquatic environment, the boundary between air-water and water-substrate, have distinctive physical characteristics that facilitate the production of communication signals. Recent evidence suggests that animals living on or near these boundaries use the interface to generate signals in 2 ways: (a) by producing a signal that propagates along the interface or (b) by producing a signal at the interface that is transmitted and detected within 1 of the component media. By examining the diversity of behaviors used to produce signals at these boundaries, the authors illustrate how human perception of these environments may cause researchers to incorrectly assume the environmental context of signal-generating behaviors and overlook modalities of communication pertinent to the animal. PMID- 12083609 TI - Atlantic bottlenose dolphin (Tursiops truncatus) hearing threshold for brief broadband signals. AB - The hearing sensitivity of an Atlantic bottlenose dolphin (Tursiops truncatus) to both pure tones and broadband signals simulating echoes from a 7.62-cm water filled sphere was measured. Pure tones with frequencies between 40 and 140 kHz in increments of 20 kHz were measured along with broadband thresholds using a stimulus with a center frequency of 97.3 kHz and 88.2 kHz. The pure-tone thresholds were compared with the broadband thresholds by converting the pure tone threshold intensity to energy flux density. The results indicated that dolphins can detect broadband signals slightly better than a pure-tone signal. The broadband results suggest that an echolocating bottlenose dolphin should be able to detect a 7.62-cm diameter water-filled sphere out to a range of 178 m in a quiet environment. PMID- 12083610 TI - Seismic sensitivity in the desert golden mole (Eremitalpa granti): a review. AB - Behavioral and anatomical studies relating to possible seismic sensitivity in the desert golden mole (Eremitalpa granti) are reviewed. Field studies in the Namib desert have shown that isolated hummocks of dune grass generate low-frequency vibrations, distinct from the background noise at a distance of many meters. The golden mole apparently uses these cues to orient itself toward the hummocks and the prey species within. An analysis of middle ear morphology suggests that the massive malleus of the golden mole is adapted toward a form of inertial bone conduction, suitable for the detection of seismic cues obtained in this manner. The significance of seismic sensitivity in this golden mole is briefly discussed. PMID- 12083611 TI - Overview: Expanding an animal's self-world to include conspecifics. AB - To further the thinking about animal communication, the authors have returned to J. von Uexkull's (1934/1957) original writings on the Umwelt. Applying J. von Uexkull's self-world concept to studies of communication helps to highlight some of the unanswered questions in behavior. The authors illustrate that some researchers have adopted views that complement and expand the ideas outlined in J. von Uexkull's classic concept of an Umwelt by exploring the influence of individual, social, and ontogenetic contexts on communication. The authors maintain that further application of these concepts expands the understanding of animal communication and, more generally, of animal behavior. PMID- 12083612 TI - Using information theory to assess the diversity, complexity, and development of communicative repertoires. AB - The application of quantitative and comparative measures from information theory on animal communication can provide novel insights into the ecological, environmental, social, and contextual properties that shape the structure, organization, and function of signal repertoires. Using 2 phylogenetically different mammalian species that share similar ecological and social constraints as examples, the authors quantitatively examined the internal structure and development of a subsystem of these species' vocal repertoires in comparison with that of human language and illustrated that these species exhibit convergent developmental processes. The authors also discussed how predictions on the structure and organization of animal communication systems can be made from this new application of information theoretic measures with respect to behavioral ecology. PMID- 12083613 TI - The presumption of sociality: social learning in diverse contexts in brown-headed cowbirds (Molothrus ater). AB - Data are presented on social and vocal learning in cowbirds (Molothrus ater) housed in large aviaries and given more degrees of freedom than in conventional experimental studies. The studies show that social and vocal outcomes are facultative responses to social contexts. Several findings are reviewed: First, cowbirds quickly self-organize into groups by age and sex; second, opportunities to interact across age and sex do exist and affect courtship competence; third, female cowbirds organize themselves differently in the presence and absence of male competition; and fourth, young, naive cowbirds show rapid and differential sensitivity to group dynamics. Taken as a whole, the data show that social Umwelten are dynamic, developmental ecologies. PMID- 12083614 TI - Multiple environmental contexts and communication in pygmy marmosets (Cebuella pygmaea). AB - There are multiple components to the concept of Umwelt experienced by an organism that may constrain the type and structure of communication signals as well as the usefulness of these signals. To illustrate the impact of these multiple environmental components, the authors used signals of the pygmy marmoset (Cebuella pygmaea), a small primate from the western Amazon. The authors summarize studies to show how the physical effects of the habitat; effects of other species, both predators and nonpredators; anthropogenic effects on the communication environment; within-group and between-groups influences other pygmy marmosets exert; and ontogeny influence the structure and usage of vocal signals. Communication within a species can be understood only in consideration of each of these contexts. PMID- 12083615 TI - A new perspective on barking in dogs (Canis familiaris). AB - The disparity in bark frequency and context between dogs (Canis familiaris) and wolves (Canis lupus) has led some researchers to conclude that barking in the domestic dog is nonfunctional. This conclusion attributes the differences primarily to genetic variation caused by domestication rather than to the influence of social environment on ontogeny. Other researchers, however, have concluded that vocal usage and response to vocalizations in mammals are strongly guided by social interactions. Closer evaluation of dog vocalizations with respect to social environment reveals developmental factors that lead to both frequent barking and barking in many contexts. Additionally, spectrographic analysis indicates that bark structure varies predictably with context, suggesting that barks can be divided into contextual subtypes and may be a more complex form of communication than given credit. PMID- 12083616 TI - Overview: For crying out loud--dangers and opportunities of communicating in the public domain. AB - Communication often extends into the public domain, inviting "eavesdropping" and other forms of exploitation by other species. Conflict is frequently the result, although interspecific communication can be mutually beneficial even when interests conflict fundamentally, such as between predator and prey. However, the most complex communication systems are likely to involve species with mutual rather than with conflicting interests. When interests do conflict, exploitation of the communication systems of others may impose high costs on signalers and select for signals that are less exploitable. Other costs of communication in the public domain arise from "mistakes" (e.g., mating with the wrong species) and select for more species-specific signal structure. The articles in this section illustrate the complexity of interspecific communication and yield important insights into the dynamic nature of communication systems. PMID- 12083617 TI - The rattling sound of rattlesnakes (Crotalus viridis) as a communicative resource for ground squirrels (Spermophilus beecheyi) and burrowing owls (Athene cunicularia). AB - Animal communication involves very dynamic processes that can generate new uses and functions for established communicative activities. In this article, the authors describe how an aposematic signal, the rattling sound of rattlesnakes (Crotalus viridis), has been exploited by 2 ecological associates of rattlesnakes: (a) California ground squirrels (Spermophilus beecheyi) use incidental acoustic cues in rattling sounds to assess the danger posed by the rattling snake, and (b) burrowing owls (Athene cunicularia) defend themselves against mammalian predators by mimicking the sound of rattling. The remarkable similarity between the burrowing owl's defensive hiss and the rattlesnake's rattling reflects both exaptation and adaptation. Such exploitation of the rattling sound has favored alternations in both the structure and the deployment of rattling by rattlesnakes. PMID- 12083618 TI - The evolution of mutualistic interspecific communication: assessment and management across species. AB - This article investigates the evolution of interspecific communication within a mutualistic setting, using the assessment-management approach to communication as a theoretical framework. The author introduces sequential stages in the evolution of mutualistic interspecific communication from unilateral assessment, to bilateral assessment, to asymmetric communication, and finally to highly complex, coevolved bilateral assessment and management and presents animal examples of each evolutionary stage. From these examples, the author forms predictions regarding when and how interspecific communication is most likely to develop, along with suggestions for further research. PMID- 12083619 TI - Species identity and the temporal characteristics of fish acoustic signals. AB - Analyses of the acoustic signals of fish show that fine-scale temporal patterns of signals are what vary among species. A growing body of research addressing the topic of species differences in fish acoustic signals suggests that these differences are related to mate choice or species isolation. However, little behavioral work has been done to determine whether these temporal differences are actually used in discriminating conspecific sounds from interspecific sounds. In this article, the authors review three cases--Centrachids, Mormyrids, and Pomancentrids--for which species specificity in both signal production and differential response to acoustic signals have been demonstrated. Work done on damselfish (Dascyllus albisella) is an especially good example and thus may serve as a model for future work. PMID- 12083621 TI - Arteriovenous fistula complicating iliac artery pseudo aneurysm: diagnosis by CT angiography. AB - Fistula formation to the inferior vena cava is a rare complication of aortic aneurysm which is often misdiagnosed clinically. In one hundred of reported arteriocaval fistulae, none was originating from the right common iliac artery. We report a case of ileo-caval fistula due to a iatrogenic pseudoaneurysm. High resolution 3D imaging using breath-hold CT angiography is highly specific in identifying the location, extent of the aortocaval fistula as well as the neighbouring anatomic structures. PMID- 12083622 TI - Lipoma in the cerebellopontine angle and internal auditory canal. PMID- 12083620 TI - Diffusion-weighted MR imaging in the evaluation of renal infection: preliminary results. AB - Diffusion-weighted (DW) magnetic resonance imaging (MRI) has already gained status in the neuroradiological MRI approach of a patient suffering from a variety of neurological diseases. The clinical application of DW MRI in the evaluation of renal disease is not standard. This manuscript describes preliminary results of the application of DW imaging in renal infection to aid differential diagnosis and/or lesion detection in clinical MRI. Three patients with acute pyelonephritis and two patients with renal abscess (one with xanthogranulomatous pyelonephritis, the other with a solitary pyogenic renal abscess) were examined with MRI. Areas of acute pyelonephritis and abscedation have restricted proton diffusion and are demonstrated on the DW images. Refinement of the technique, in-depth investigation of the pathological background of the MR findings and evaluation of its true clinical value need further investigation. This manuscripts shows preliminary findings of DW imaging in patients with renal infectious disease. PMID- 12083623 TI - Pseudocoarctation or aortic kinking. PMID- 12083624 TI - How to implement global integration between EPR and imaging. The Mont-Godinne's experience. PMID- 12083625 TI - Role of magnetic resonance imaging in coronary heart disease. AB - Coronary heart disease (CHD) is the number one cause of morbidity and mortality in the developed countries. CHD involves a very broad clinical spectrum, ranging from stable angina pectoris to sudden cardiac death. Despite a huge improvement in treatment in the last 3 decades, significantly reducing the death rate, the prevalence of CHD will increase in the coming years due to an improved survival after a first acute myocardial infarction and due to an overall increased longevity, fueled by an epidemic of type-2 diabetes and obesity. This increased prevalence of cardiovascular disease will greatly impact the cost of health care. Moreover, new but often expensive imaging technologies are appealing for more accurate clinical and preclinical detection of CHD. Among these techniques, magnetic resonance imaging (MRI) is certainly one of the most promising. In this review we would like to highlight its potential in diagnosing CHD, against the background of a steadily increasing cost in health care and the need to choose the most cost-effective technique. PMID- 12083626 TI - Blood vessel invasion as a predictor of long-term survival for Japanese patients with breast cancer. AB - A wide range of frequencies has been reported for blood vessel invasion (BVI) among patients with breast cancer, however, the prognostic significance of BVI remains controversial. Three hundred ninety-eight Japanese patients with breast cancer, operated on during the period between 1971 and 1987, were studied. We investigated five factors, including BVI, lymph-node status (n), clinical tumor size (T), histological grade (HG), and tumor necrosis (TN), followed for a median of 10 years. BVI was detected by hematoxylin and eosin (HE) staining and both factor VIII-related antigen and elastica van Gieson staining. BVI detected by HE staining alone was defined as BVIh. The subtypes of BVI were classified as follows: BVI e, BVI detected only by elastica van Gieson staining; BVI f, BVI detected only by factor VIII-related antigen staining; and BVI e/f, BVI detected by both factor VIII-related antigen and elastica van Gieson staining. BVI positive tumors were defined as lesions showing BVI e, BVI f, or BVI e/f. BVI and BVIh were presented in 27.4%, 6.5% of all cases, respectively. The mean diameters of the calibers of BVI e, BVI f, and BVI e/f were 141.9+/-80.5 microm, 61.0+/ 37.4 microm, 136.0+/-102.0 microm, respectively (P<0.0001). Seventy-three patients (18.3%) had recurrence and 60 patients (15.1%) died of breast cancer. Univariate analysis showed that BVIh (P <0.0001), BVI (P<0.0001), n, T, and HG were significantly predictive of 20-year RFS and OS. Multivariate analysis showed that BVI (P<0.0001, P = 0.0088, respectively), n, T, and HG were all significant and independent prognostic factors for RFS and OS. On the other hand, BVIh was an independent factor for RFS (P = 0.0475), but of borderline significance for OS (P = 0.0506). When stratified by BVI, BVI e, and BVI e/f were significantly predictive of 20-year RFS or OS (P < 0.0001). We can confirm BVI, especially BVI e and BVI e/f, are significant independent prognostic factors associated with long-term survival in Japanese breast cancer patients. PMID- 12083627 TI - The influence of the route of administration of 17beta-estradiol, intravenous (pulsed) versus oral, upon DMBA-induced mammary tumour development in ovariectomised rats. AB - A wide variety of routes of administration and formulations are employed in estrogen replacement therapy. These exhibit differences in the pharmacokinetics and metabolism of estradiol and in the resulting biological effects. This study set out to investigate the effects of pulsed estrogen administration (via the nasal route) compared to oral therapy, as a reference, with regard to breast cancer risk. This was assessed in an experimental model whereby mammary tumours were induced by 7,12-dimetbylbenz(a)anthracene in ovariectomised rats. To mimic a pulsed treatment given via the nasal route doses of estrogen were administered by I.V. route (0.4, 10 and 250 microg/kg). These dosages were predicted to have similar estrogenic activity to doses administered by the oral route (100, 300 and 900 microg/kg). Controls were groups of ovariectomised and SHAM-operated rats and ovarectomised rats administered with either vehicle alone. Two studies were carried out on separate populations of rats and ran in parallel. Tumour appearance (study 1) and tumour growth (study 2) were evaluated. In study I (n = 20/group), treatments with estradiol were conducted for 20 weeks after carcinogen administration; in study 2 (n = 10/group), an 8-week treatment with estradiol was initiated once 7,12-dimethylbenz(a)anthracene-induced tumours appeared. Intravenous dose levels achieved equivalent estrogenicity to corresponding oral dose levels, as assessed by measuring uterus weight. Estrogen deficit was made up by both routes but only the higher doses restored physiological uterus weight. Nevertheless administration via the I.V. route resulted in a lower rate of tumour incidence (p < or = 0.05) than the rate recorded for the oral route. In addition, tumour development was lower with the I.V. route. In conclusion, in this experimental model, pulsed estrogen therapy with 17beta-estradiol administered via the I.V. route resulted in a reduced effect on mammary carcinogenesis when compared to oral administration. PMID- 12083628 TI - Long-term effects of the mammary carcinogen 7,12-dimethylbenz(a) anthracene on hypothalamic gonadotropin-releasing hormone and its pituitary receptor gene expression, during the promotion stage, in female Sprague-Dawley rats. AB - A single intragastric administration of 7,12-dimethylbenz(a)anthracene (DMBA) has been shown to induce mammary tumors in young cycling female Sprague-Dawley rats. The appearance of these tumors is preceded by a series of neuroendocrine disturbances, including attenuation of the preovulatory luteinizing hormone (LH) surge and amplification of the preovulatory 17beta-estradiol surge, and gonadotropin-releasing hormone (GnRH) released in vitro. In this study, we examined the hypothesis that DMBA administration decreases levels of GnRH mRNA in the preoptic area-anterior hypothalamus (POA-AH) and GnRH receptor (GnRH Rc) mRNA and protein in the anterior pituitary gland. Sprague-Dawley rats, 55-60 days of age with regular estrous cycles, received a single dose of 15 mg DMBA in 1 ml sesame oil delivered by intragastric intubation. A first series of experiments was performed for the measurement of hypothalamic GnRH mRNA and pituitary GnRH Rc mRNA levels. A second series of experiments was performed for the measurement of pituitary GnRH receptor. In both experiments, animals were sacrificed by decapitation at 11.00, 16.00, 18.00 and 20.00 h on each day of the 7th or 8th estrous cycle (28-32 days) after treatment. GnRH and GnRH receptor mRNAs were quantified using solution hybridization-RNase protection assay. The GnRH Rc was quantified using the 125I-D-Ala6-N-Met-Leu6-des-Gly10-ethylamide GnRH. DMBA treatment produced no significant effect on the overall mean values of GnRH mRNA. GnRH mRNA levels in control rats rose significantly between 16.00 and 20.00 h on proestrus and between 18.00 and 20.00 h on diestrus I. DMBA-treated rats had a surge in GnRH mRNA levels at 18.00 h on proestrus, and showed additional surges at 18.00h on diestrus II and estrus. GnRH receptor mRNA content in the anterior pituitary gland surged at 16.00h on certain days of the cycle in both groups of rats. In control rats, only the surge on diestrus II proved significant, whereas DMBA-treated rats exhibited significant surges on diestrus I, diestrus II and proestrus. GnRH receptor mRNA values were significantly lower on both days of diestrus in DMBA-treated rats compared with controls. GnRH Rc peptide content, like GnRH receptor in RNA surged at 16.00h in both groups with the exception of a marked fall on proestrus day for DMBA treated rats. A reduction in the amplitude of the surge was also seen on the day of estrous and to a lesser extend on the day of diestrus DII in DMBA treated animal. Overall, there was a disruption of the GnRH Rc pattern which culminate on the day of proestrus in DMBA-treated animals. Interestingly, the daily rise between 11.00 and 16.00h which is the more pronounced on the day of proestrus in control animals, was completely blunted in DMBA-treated rats. Overall, the results are consistent with the hypothesis that the carcinogen attenuates, directly or indirectly, preovulatory biosynthesis of the GnRH receptor and LH release. Obviously, the changes in GnRH might occur simultaneously, independently from mammary tumorigenesis, but may play a role, in association with others DMBA-induced neuroendocrine disorders, in the promotion stage of mammary tumors in the Sprague-Dawley female rat. PMID- 12083629 TI - Morbidity of breast cancer patients following complete axillary dissection or sentinel node biopsy only: a comparative evaluation. AB - INTRODUCTION: The usefulness of routine axillary dissection (AD) at levels I-II in breast cancer patients has been questioned for years because of the high postoperative morbidity in the shoulder and arm region, and the increasing number of patients with negative nodes. Sentinel node biopsy (SNB) was hoped both to reduce morbidity and to improve the reliability of staging. This study was designed to provide more evidence in this matter by comparing the follow-up data of patients with AD and those with SNB only. METHOD: One hundred forty patients who had undergone AD between 1993 and 1996 were questioned for their subjective and objective symptoms using a questionnaire and subsequently subjected to a clinical examination. Their data were compared with those of 57 patients who had undergone SNB only between 1998 and 2000. RESULTS: Local recurrences have not been seen to date. The difference between the two groups in terms of a loss of quality of life was negligible. The differences in overall complaints, number of symptoms, pain, limited range of motion of the operated upper extremity, numbness, paresthesias, and arm swelling as well as perceived disability in activities of daily living were significantly in favor of SNB. The length of hospital stay was significantly shorter for SNB patients. CONCLUSION: SNB appears to be an accurate procedure for axillary nodal staging in breast cancer patients and is associated with reduced postoperative morbidity and length of hospital stay. But it is still investigational and should not be implemented as therapeutical standard before results of randomized trials are published. PMID- 12083630 TI - Impact of videotaped information on frequency and confidence of breast self examination. AB - BACKGROUND: Videotaped education materials to teach breast self-examination (BSE) are used worldwide. However, evaluation of videotaped BSE instructions is lacking. METHODS: Premenopausal women (mean age 33.4+/-11.2 years) without history of breast cancer were approached to participate in this experimental study and randomly assigned to a video intervention group (VG; n = 130: length of the video = 15 min) or non-video comparison group (NVG; n = 121). All participants answered a questionnaire on BSE behavior and health beliefs. No additional training was given. The total duration of the session including completion of the questionnaire was 15 min for the NVG and 30 min for the VG. Three months later, changes in BSE behavior were compared in the two groups. The influence of health beliefs on actual BSE behavior was investigated as well. RESULTS: Women of both the VG and NVG performed BSE significantly more frequently at follow-up than at baseline. Analysis of covariance, using the baseline BSE frequency as co-variate and the follow-up BSE frequency as the dependent variable, revealed that women in the VG (adjusted mean = 7.9 times per year, 95%CI = 6.5-9.4) performed BSE more frequently than women of the NVG (adjusted mean = 6.1 times per year, 95%CI = 4.6-7.5) (F = 4.2, df= 2, p = 0.02). Among motivational predictors, having an example of a role model (modeling) was shown by regression analysis to explain the greatest amount of variance (13%) in BSE frequency. CONCLUSION: Use of an educational videotape increased the frequency of BSE among premenopausal women. PMID- 12083631 TI - Hormone receptors in breast cancer: racial differences in distribution and survival. AB - The purpose of this study was to describe hormone receptor status and analyze the effect of receptors on survival from breast cancer. Comparisons were made between African-American and Caucasian racial categories. Breast cancer data from 1990 through 1997 collected by the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) Program were analyzed. Subjects were 993 Caucasian men, 12,303 African-American women, and 141,045 Caucasian women. The number of African-American men was too small to analyze separately (n = 93). In addition to analysis of estrogen and progesterone receptor status by sex and race, tumor and patient characteristics included age, stage at time of diagnosis, and tumor histology. The proportion of Caucasian men with hormone receptor positive tumors remained relatively high and stable for all ages. In women, the proportion of hormone receptor positive tumors increased with age, with African American women having the highest proportion of hormone receptor negative tumors. Caucasian men had highest proportions of hormone receptor positive tumors in all histology and stage groups, while African-American women had lowest proportions of hormone receptor positive tumors in all stage and histologic categories. Survival for African-American women was significantly worse for each hormone receptor category. In multivariate analyses, race was a significant independent predictor of survival, but sex was not. Although reasons for differences in hormone receptor status by sex and race are unknown, several hypotheses are discussed with respect to differences in tumor histopathology and risk factors. PMID- 12083633 TI - Effect of selective ablation of proliferating mammary epithelial cells on MNU induced rat mammary tumorigenesis. AB - Proliferating cells within the terminal end buds of the virgin female rat mammary gland are the most susceptible to chemical carcinogen induced tumorigenesis. We hypothesized that selective ablation of proliferating cells in the mammary gland would reduce mammary tumor incidence upon carcinogen challenge. Selective ablation of proliferating cells was achieved by intraductal injections of Adv-RSV tk and gancyclovir administration. Despite efficient viral transduction of the thymidine kinase protein and the apparent elimination of >90% of the proliferating cells, the rats exhibited a higher incidence of MNU induced mammary tumors arising with shorter latency as compared to control animals. Several possible explanations of the puzzling relationship between elimination of cycling cells and increased tumor incidence are discussed and alternative strategies for the prevention of breast cancer are proposed. PMID- 12083632 TI - Arsenic trioxide and breast cancer: analysis of the apoptotic, differentiative and immunomodulatory effects. AB - Arsenic trioxide (As2O3) is used clinically to treat acute promyelocytic leukemia and has activity in vitro against several solid tumour cell lines, where induction of differentiation and apoptosis are the prime effects. To investigate the potential therapeutic application of As2O3 to breast cancer, we analysed the effects of As2O3 on the growth of four human breast cancer cell lines: MCF7, MDA MB-231, T-47D and BT-20. Cells were cultured in 0.5, 2 and 5 microM AS2O3, a range of pharmacologically achievable concentrations of AS2O3. At > or = 2 microM, AS2O3 rapidly induced cell death by apoptosis in MCF7 and MDA-MB-231 while T-47D and BT-20 were partially resistant. At 0.5 microM, As2O3 was subapoptotic but induced features of differentiation consisting in upregulation of ICAM-1 (CD54), a marker of mammary epithelial differentiation, and cell cultures appeared morphologically more organized. Furthermore, we demonstrate by standard cytotoxicity assays that As2O3 treatment can augment breast cancer cell lysis by lymphokine-activated killer cells and demonstrate an important role of the ICAM-1/LFA-1 interaction in this process. This additional activity of As2O3 could translate into improved antitumour immunosurveillance in vivo. In conclusion, As2O3 induced varying degrees of differentiation, apoptosis and lysis in these model cell lines, and may be a promising adjuvant to current treatments of breast cancer by virtue of its triple apoptotic, differentiative and immunomodulatory effects. PMID- 12083634 TI - Weekly paclitaxel in women age 65 and above with metastatic breast cancer. AB - We evaluated therapy with weekly paclitaxel 80 mg/m2 in metastatic breast cancer patients age > or =65. There was a low incidence of serious toxicities, with similar tolerability profiles in younger and older patients. Response rates and overall survival times were comparable in the two age groups (<65 and > or =65). Weekly paclitaxel therapy is a reasonable option for older patients with metastatic breast cancer. PMID- 12083635 TI - Re: Mammography utilization among California women aged 40-49 in a managed care environment. PMID- 12083636 TI - Ah, that new car smell. Finding your personal style for dealing with change. PMID- 12083637 TI - Brush biopsy 'saves lives'. PMID- 12083638 TI - Brush biopsy 'bridges the gap'. PMID- 12083639 TI - 'Overwhelmingly positive'. PMID- 12083640 TI - Abfraction. PMID- 12083641 TI - JADA'S industry advisory board. PMID- 12083642 TI - Working with industry. PMID- 12083643 TI - Injection concerns. PMID- 12083644 TI - Injection complications. PMID- 12083645 TI - Do you think a cure for AIDS will be found in your lifetime? PMID- 12083646 TI - The impact of a state children's health insurance program on access to dental care. AB - BACKGROUND: Access to dental care for low-income children is limited. The authors examined the impact of a new state children's health insurance program, or SCHIP, in North Carolina on children's access to dental care. METHODS: Parents of 639 school-aged children responded to two surveys that asked about their child's access to dental services before enrollment and one year after enrollment in the new program. The authors used two-tailed McNemar tests to detect statistically significant changes within subjects. RESULTS: The percentage of school-aged children with a visit to a dentist in the previous year increased from 48 percent at baseline to 65 percent after one year in the program. Reported unmet dental need decreased from 43 percent at baseline to 18 percent after one year of enrollment. The proportion of children reported to have a usual source of dental care improved after enrollment in the program. CONCLUSION: The SCHIP model in North Carolina is an innovative program that has made a significant impact on access to dental care for school-aged children. PRACTICE IMPLICATIONS: SCHIP dental programs that resemble private insurance models and reimburse dentists at rates close to market rates hold the potential to address problems associated with dental access for low-income children. PMID- 12083647 TI - Dental visits to hospital emergency departments by adults receiving Medicaid: assessing their use. AB - BACKGROUND: Pain from toothaches represents a significant problem. People lacking access to private dental services may use hospital emergency departments, or EDs. In 1993, Maryland eliminated Medicaid reimbursement to dentists for adult emergency services. METHODS: The authors used the change in Medicaid policy that eliminated dentist reimbursement to establish two study periods. Data tapes describing patients' use of EDs were obtained from the Maryland Medicaid Management Information System. A total of 3,639 people visited EDs for dental problems sometime during the four-year study period. RESULTS: After controlling for age, race and sex, the authors found that the rate of ED claims was 12 percent higher in the postchange period than in the prechange period. Comparisons between periods show significant rate increases during the postchange period for men, whites, African-Americans and patients aged 21 through 44 years and 45 through 64 years. CONCLUSIONS: The change in Medicaid policy that eliminated dentist reimbursement and participation in the program appears to have increased the use of EDs for the treatment of dental problems. Practice Implications. Many EDs lack dental services and are not capable of providing definitive treatment. When definitive treatment is not provided, this pattern of care may be repeated if patients are forced to return for treatment. PMID- 12083648 TI - Characteristics of noncarious cervical lesions: a clinical investigation. AB - BACKGROUND: The purpose of the authors' in vivo investigation was to analyze the characteristics of noncarious cervical lesions, or NCCLs, in adult patients who had a high incidence of them. METHODS: The patient pool consisted of a total of 57 patients and 171 teeth (three teeth per patient), with one NCCL per tooth. The characteristics the authors evaluated were shape, dimensions, sensitivity, sclerosis and occlusion. RESULTS: In terms of lesion characteristics, 91 percent of the lesions had axial depths of 1 to 2 millimeters, 49 percent had occlusogingival widths of 1 to 2 mm, 74 percent had an angular shape of 45 to 135 degrees, 76 percent had mild or moderate sclerosis, and 73 percent had no or mild sensitivity. In terms of occlusion, 75 percent of teeth had an Angle Class I occlusion on the involved side, 60 percent had group function or mixed excursive guidance, 82 percent had wear facets, and 99 percent had Type 0 or I mobility. In terms of tooth location, 70 percent of NCCLs were on posterior teeth, 65 percent were on maxillary teeth, and 46 percent were on premolars. CONCLUSIONS: The evaluated NCCLs were found mainly to have small dimensions of depth and width (< 2 mm) and to be roughly right-angled in shape, and many had sclerosis and low sensitivity. A majority of the dentitions studied had Class I occlusion, with group function, prevalent wear facets, and little or no mobility. Cervical lesions were more common with posterior maxillary teeth and premolars, especially first premolars, which had the highest prevalence of lesions. Older patients were more likely to exhibit noncarious cervical lesions, but no great difference in incidence was found between men and women. Clinical Implications. A knowledge of the NCCL characteristics and etiologic covariables aids in proper case selection for treatment, aids in selection of appropriate treatment protocols and improves assessment of prognosis. PMID- 12083649 TI - Dental erosion caused by silent gastroesophageal reflux disease. AB - BACKGROUND: Gastroesophageal reflux disease, or GERD, is a relatively common condition, in which stomach acid may be refluxed up through the esophagus and into the oral cavity, resulting in enamel erosion. Symptoms such as belching, unexplained sour taste and heartburn usually alert the patient to the condition. In silent GERD, however, these symptoms do not occur, and enamel erosion of the posterior dentition may be the first indication of GERD. CASE DESCRIPTION: A 30 year-old man came to a dental clinic with enamel erosion on the occlusal surfaces of his posterior teeth and the palatal surfaces of his maxillary anterior teeth. He reported no history of gastrointestinal disease or heartburn. CLINICAL IMPLICATIONS: Enamel erosion may be a clinical sign of silent GERD that allows the dentist to make the initial diagnosis. Referral to a physician or gastroenterologist is necessary to define the diagnosis; however, dental expertise may be essential in distinguishing between differential diagnoses such as bulimia, attrition and abrasion. Successful treatment of this medical condition is necessary before dental rehabilitation can be initiated successfully. PMID- 12083650 TI - Bilateral parotid swelling caused by sarcoidosis. AB - BACKGROUND: Sarcoidosis is a multisystem granulomatous disease characteristically causing bilateral hilar lymphadenopathy. Salivary gland swellings also are seen, which mandate recognition by the dental profession. CASE DESCRIPTION: The authors describe a patient whose diagnosis of sarcoidosis was tenuous. Ocular involvement and an elevated serum angiotensin-converting enzyme level suggested a diagnosis of sarcoidosis. Confirmation with a tissue biopsy was achieved only when the clinicians recognized that salivary gland swelling probably represented a manifestation of the disease. Steroid therapy was instituted. CLINICAL IMPLICATIONS: It is important for the dental practitioner to be familiar with the salivary gland manifestation of sarcoidosis. Early recognition and intervention aborts or slows disease progression. PMID- 12083652 TI - A new perspective on dental malpractice: practice enhancement through risk management. AB - BACKGROUND: Risk management in dentistry has been developed over the years by concentrating on recording treatment in dental records and informing patients of the proposed treatment before treating them. This article advances the concept of risk management through higher involvement of the entire office staff by increasing communication with patients. CONCLUSIONS: By integrating practice management concepts with risk management techniques, dentists can reduce risk management exposure and improve patients' awareness, understanding and follow through on the treatment of their dental needs. PRACTICE IMPLICATIONS: Practice enhancement through risk management not only improves patient care and reduces risk exposure, but it also brings the dental team together in an effort to improve patient care. In this way, the office will have improved patient acceptance of proposed dental care and an increase in office growth. PMID- 12083651 TI - Preventing the negative sequelae of tooth extraction. PMID- 12083653 TI - Biocompatibility of dental alloys. PMID- 12083654 TI - The curing light dilemma. AB - At this time, all the light-curing technologies on the market work adequately when used as directed by the manufacturers. The trend in the profession is to accept faster curing of resin-based composite. I agree completely. I have been using fast curing lights for many years, and I do not see any scientific or clinical reasons to change. PMID- 12083655 TI - Hawaii court dismisses antitrust suit against provider groups. PMID- 12083656 TI - For the dental patient. Dental emergency procedures. PMID- 12083657 TI - Vibrational frequencies and structural determinations of hexamethylenetetraamine. AB - The normal mode frequencies and corresponding vibrational assignments of hexamethylenetetraamine (HMTA) in Td symmetry are examined theoretically using the Gaussian 98 set of quantum chemistry codes. All normal modes were successfully assigned to one of eight types of motion predicted by a group theoretical analysis. The vibrational modes of the deuterated form of HMTA (HMTA d-12) were also calculated and compared against experimental data. The normal mode vibrational frequencies were shifted to lower frequencies as on deuteration as expected. However, in some cases the dominant motion type changed on deuteration leading to an apparent 'blue shift' of some of the N-C stretching modes. It is possible that the observed frequency shifts are the result of a Fermi resonance condition. PMID- 12083658 TI - Stereochemistry, structural and models of novel 5-(4'-derivatives phenyldiazo)-3 phenyl-2-thioxo-4-thiazolidinone complexes. AB - Copper(II) and cadmium(II) complexes of 5-(4'-derivatives phenyldiazo)-3-phenyl-2 thioxo-4-thiazolidinone (HLn) were prepared, their compositions and physicochemical properties were characterized by elemental analysis, magnetic suseptibility measurements, and infrared, electronic spectra. The novel complexes have the stoichiometric formulae [Cu(HLn)(OAc)n(H2O)(X)] (OAc = acetate, X = H2O or acetate) and [Cd(L)(OAc)(H2O)], respectively. Elemental analysis and IR spectra denote, that two types of complexes with different octahedral and tetrahedral structure for Cu(II) and Cd(II) ions. I.R. spectra show that the ligand is monobasic/neutral bidentate forming thereby a six-membered chelating ring and concomitant formation of an intramolecular hydrogen bond. The stoichiometeries of these complexes were determined conductometrically and indicated the formation of 1:1 and 1:2 (metal:ligand) complexes. PMID- 12083659 TI - Spectrophotometric study of molecular complex formation between o-chloranil and a series of methylated pyridines. AB - o-Chloranil has been shown to form 1:1 molecular complexes with pyridine and 2-, 3- and 4-picolines in CCl4 medium. Isosbestic points have been found but charge transfer bands could not be detected. The formation constants of the complexes exhibit a very good linear free energy relationship from which the Hammett p parameter for the complexation reaction is found to be -3.67. PMID- 12083660 TI - Optical properties of Nd3+ ions doped in oxyfluoroborate glass. AB - Laser induced excitation spectrum of the Nd3+ ions doped in oxyfluoroborate glass have been recorded. Stark components of different electronic states have been reported. Judd-Ofelt analysis has also been accomplished on the basis of ultraviolet-visible-near-infrared (UV-vis-NIR) absorption spectrum. Various radiative parameters such as electric dipole line strength, transition probability, life time of the various energy levels, branching ratio etc., have been estimated. PMID- 12083661 TI - The spin population difference of hydrogen atoms dissociated from mercury hydride molecules. AB - The spin population difference between m(s) = 1/2 and - 1/2, (malpha - mbeta)(t) of the hydrogen atoms dissociated from the mercury hydride molecules is calculated as a function of time for the reaction of Hg* + H2 in a magnetic field. Starting from zero value, the population difference increases to a certain limit that is equal to the room temperature population difference. The H/D electron spin resonance (ESR) signal ratio obtained from the isotopic experiment, Hg* + H2/D2, is also explained by comparing the ratio of (malpha - mbeta)H(t) and (malpha - mbeta)D(t). The isotopic effect is interpreted to originate mostly from spin-rotation constant energies of mercury hydride molecules. PMID- 12083662 TI - Vibrational investigation of the stretching region of bromate ion in solution. AB - The splitting of v1 (794 cm(-1)) and v3 (805 cm(-1)) modes in the stretching region of the bromate (C3v) was observed for the first time in dimethyl sulfoxide (DMSO) using vibrational spectroscopy. Depolarization measurements allowed to assign the asymmetric and symmetric modes in solution. The band at 805 cm(-1) that has been attributed to the symmetric stretching mode (A1) corresponds in fact to the asymmetric stretching mode (E) and the band at 794 cm(-1) corresponds to the symmetric stretching mode (A1). PMID- 12083663 TI - Spectrophotometric studies on the simultaneous determination of cadmium and mercury with 4-(2-pyridylazo)-resorcinol. AB - A new method for direct spectrophotometric determination of cadmium with 4-(2 pyridylazo)-resorcinol is reported. Absorption maximum, molar absorptivity and Sandell's sensitivity of the 1:1 (M:L) complex are 510 nm, 2.5 x 10(5) l mol(-1) cm(-1) and 3.55 ng cm(-2), respectively. A linear calibration graph is obtained up to 4.49 microg ml(-1). The zero-crossing measurement technique is found suitable for the direct measurement of the first-derivative value at the specified wavelengths. Cadmium(II) (0.42-9.2 microg ml(-1)) and mercury(II) (0.35 7.4 microg ml(-1)) in different ratios have been determined simultaneously. A critical evaluation of the proposed method is performed by statistical analysis of the experimental data. The developed method was applied to the simultaneous spectrophotometric determination of Cd and Hg in some synthetic mixtures and was found to give satisfactory results. PMID- 12083665 TI - Multinuclear 1H, 13C and 15N NMR study of some substituted 2-amino-4 nitropyridines and their N-oxides. AB - 1H, 13C and 15N NMR chemical shift assignments based on pulsed field gradient selected PFG 1H,X (X = 15C and 15N) HMQC and HMBC experiments are reported for three 4-nitropyridine N-oxides and four 4-nitropyridines. It was found that an ortho effect of a methyl group inhibits the deshielding effect of the 4-nitro group and that this effect and the so-called back donation is influenced by electronegativity and position of substituents in the multisubstituted pyridine N oxides. The shielding effect of N-oxide group is most pronounced in the 15N NMR chemical shifts of the studied compounds. This effect is further modified by methylamino, methylnitramino, 5- or 3-methyl and 4-nitro groups. Among them the 4 nitro group exerts the highest influence on the shielding effect of the N-oxide functionality. Experimental 1H, 13C and 15N NMR chemical shifts and GIAO/DFT theoretical calculations are consistent with each other and supported by the reactivity on nucleophilic substitution, the UV spectral and the dipole moment data. PMID- 12083664 TI - Polymer complexes XXXVII novel models and structural of symmetrical poly-Schiff base on heterobinuclear complexes of dioxouranium(VI). AB - Some binary and ternary novel complexes of dioxouranium(VI) with 5 vinylsalicylaldehyde (VSH) have been prepared and characterized by various physico-chemical techniques. The amine exchange reactions of coordinated poly Schiff bases in these complexes have been also carried out which give symmetrical tetradentate poly-Schiff base complexes. Metal exchange reaction of these dioxouranium(VI) complexes with copper(II) gives the corresponding Cu(II) complexes. Reaction of tetradentate poly-Schiff base complexes of Cu(II) so obtained with ZrCl4 gives heterobinuclear polymer complexes. Magnetic, electronic and IR spectral information commensurate that configurations of square planar copper(II) polymer complexes. All the polymer complexes are coloured and appear to be nonelectrolytes in DMF. The ligands behave as bi-(O, O) and tetradentate (N2, O2) donors. El-Sonbati equation was used to evaluate the symmetric stretching frequency from which the fU-O and fUO, UO- were calculated. PMID- 12083666 TI - Solvent effects on OH stretching frequencies for 1-arylallyl alcohols. AB - A series of 1-arylallyl alcohols were prepared and its OH stretching frequencies measured in 20 different non-HBD solvents at room temperature. It is noticed that the observed stretching bands were highly sensitive to the nature of the solvents. Multiple parameter equations were applied to investigate the solvent effects on the O-H stretching frequency. The most significant solvent parameters were the nucleophilicity measuring parameter (B) and Gutmann donor number (DN), whilst the electrophilicity measuring parameter (E) is not significant. PMID- 12083667 TI - Solvent effects and photophysical studies of a new class of acridine(1,8)dione dyes. AB - Spectroscopic properties of a new family of acridinedione dyes are reported. The absorption and fluorescence spectra of the different substituted acridinediones have been recorded in different solvents and the difference in the dipole moment between ground and excited state has been obtained by solvatochromic shift method. The value of the Onsager cavity radius was calculated from the total surface area using software PCMODEL. Fluorescence quantum yield and fluorescence lifetime were determined. Radiative and non-radiative constants have been calculated. The triplet-triplet absorption maxima and triplet lifetime show variation depending on the substitution. PMID- 12083668 TI - Identification of unknown surfactants using electrospray mass spectrometry and NMR spectroscopy preceded by liquid ionization mass spectrometry. AB - Commercially available but completely unknown surfactants used in the tin-lead plating industry were successfully identified by using electrospray mass spectrometry and NMR spectroscopy techniques, preceded by liquid ionization mass spectrometry used to obtain the preliminary information. The mass spectral data suggested that ethoxylated nonionic surfactants having a homologous distribution of molecular weights like 520, 564, 608, 652, 696, etc. were present. The NMR data suggested the presence of two aromatic rings and a quaternary carbon for the hydrophobe moiety instead of the well-known alkyl chains or alkylphenols. The unknown surfactants were finally concluded to be novel nonionic 4-(alpha,alpha dimethylbenzyl)-phenol ethoxylates. PMID- 12083669 TI - Vibrational frequencies and structural determinations of 3,5-dibromo-1,2,4 trithia-3,5-diborolane. AB - The vibrational frequencies and corresponding normal mode assignments of 3,5 dibromo-1,2,4-trithia-3,5-diborolane (B2S3Br2) are examined theoretically using the Gaussian98 set of quantum chemistry codes. All normal modes were successfully assigned to one of six types of motion predicted by a group theoretical analysis (B-S stretch, B-Br stretch, S-S stretch, S-B-S bend, B-Br wag, B(SSBr) umbrella motion) utilizing the C2v symmetry of the molecule. The vibrational modes of the naturally isotopically substituted (1-10B and 2-10B) forms of B2S3Br2 were also calculated and compared against experimental data. The molecular orbitals of B2S3Br2 are examined. The calculations suggest that a considerable amount of pi bonding occurs in B2S2Br2. PMID- 12083670 TI - EPR and optical absorption studies of Cu2+ doped L-arginine phosphate monohydrate single crystals--part II. AB - The EPR spectra of Cu2+ ion in L-Arginine phosphate monohydrate (LAP) at X-band frequencies at room temperature reveal the presence of two magnetically inequivalent interstitial Cu2+ sites in the lattice. The principal values of the g- and A-tensors indicate rhombic symmetry around the Cu2+ ion. From the direction cosines of the principal values of the g- and A- tensors, the orientations of the sites in the lattice have been identified. The bonding parameters were evaluated and the ground state wave function of Cu2+ ion was constructed. The ground state is found to be an admixture of [x2 - y2> and [3z2 - r2> orbitals. The optical absorption studies show four bands centered at 9803, 10753, 15748 and 16666 cm(-1) confirming the rhombic symmetry around the Cu2+ ion. Using the observed bands the crystal field parameters have been evaluated. PMID- 12083671 TI - Electronic structure of coumarins. AB - The electronic structure of coumarin derivatives has been investigated by a combination of UV photoelectron spectra (UPS), semi-empirical MO calculations and comparison with the spectra of related coumarins. The influence of substituents on the S0 and S1 electronic states energies is discussed. PMID- 12083672 TI - The interaction of Cu2 + ions and NaDC micelles. AB - By mixing an aqueous solution of CuCl2 with an NaDC aqueous solution of various concentration and initial molar ratio, seven coordinated samples with distinct appearances and characters were obtained. Their structures and components were investigated by FT-IR spectroscopy, EXAFS (the extended X-ray absorption fine structure), thermal analysis, X-ray diffraction, laser light scattering, TEM (transmission electron micrograph), element analysis and ICP (inductively coupled plasma) analysis. The following conclusions were given: (1) The complexes of Cu2+ NaDC with distinct appearances and properties were synthesized. (2) After Cu(DC)2 dissolved in NaDC aqueous solution, larger micelles (30-90 nm diameter) formed in the supernate, it is a mixed micelle with Cu(DC)2 and NaDC. So these micelles are a new kind of micelle containing two kinds of metal ions. This is a new result using metal ions as bridges to form micelle. (3) According to the different concentration of Cu2+ to NaDC, the complexes formed as gel or poly-crystals. Both the composition of gel complexes and the coordination structure of carboxyl groups with metal ions varied with the initial molar ratio of Cu2+ to Na+. The gel complexes exhibits the non-stoichiometric character. (4) These results are in agreement with physiological condition. All the different states such as gel, precipitate, micelles of various structures are present in bile of gallbladder. We can suggest an ideal model of the interaction between Cu2+ and bile salts in vivo. PMID- 12083673 TI - Matrix isolation infrared spectroscopic study of the interaction of CH3ReO3 with NH3. AB - The matrix isolation technique has been combined with infrared spectroscopy to identify and characterize the product of the codeposition of CH3ReO3 with NH3 into inert matrices at 14 K. This codeposition led to the formation of the isolated 1:1 complex between these two reagents and its isolation in argon and nitrogen matrices. The complex is characterized by perturbations to all of the vibrational modes of the NH3 subunit in the complex, including a large, 185 cm( 1) blue shift of v2, the symmetric deformation mode. In addition, shifts of the ReO3 antisymmetric stretch and -ReO3 symmetric bending of the CH3ReO3 subunit in the complex were observed. This complex, while predicted theoretically, has not been reported previously. PMID- 12083674 TI - Single crystal EPR of Cu(II) doped Cd(C5H5NO)6(BF4)2--an example of reduced metal hyperfine coupling constant. AB - Electron paramagnetic resonance (EPR) studies have been carried out on Cu(II) doped Cd(C5H5NO)6(BF4)2 in single crystal and powder forms at 300 K. Angular variation of hyperfine structure lines shows the presence of two magnetic sites, indicating possible twinning in the Cd lattice. From the mean values of two sites, the angular variation plot is fitted to an orthorhombic spin Hamiltonian. The observed orthorhombic g values and the very low A11 value ([A11] = 107.8 x 10(-4) cm(-1)) are unusual for a hexakis coordinated system. This unique behavior is explicable in terms of the static Jahn-Teller (JT) effect. PMID- 12083675 TI - Optical emission spectroscopy of nitrogen species and plasma plume induced by laser ablation combined with pulse modulated radio-frequency discharge. AB - Combined laser ablation and pulse modulated radio-frequency (RF) discharge for deposition of CNalpha films was studied by the use of optical emission spectroscopy. Chemically active nitrogen was produced by RF discharge, concentrated between two small electrodes. Influence of RF power, nitrogen pressure, modulation frequency and pulse rate on nitrogen species production was researched. For the same system plasma expansion rate, kinetic energy and concentration of carbon ions emitted by laser from graphite target were determined by Langmuir probes measurement. PMID- 12083676 TI - Attenuated total reflection Fourier transform infrared spectroscopy of redox transitions in photosynthetic reaction centers: comparison of perfusion- and light-induced difference spectra. AB - Chemically induced Fourier transform infrared difference spectra associated with redox transitions of several primary electron donors and acceptors in photosynthetic reaction centers (RCs) have been compared with the light-induced FTIR difference spectra involving the same cofactors. The RCs are deposited on an attenuated total reflection (ATR) prism and form a film that is enclosed in a flow cell. Redox transitions in the film of RCs can be repetitively induced either by perfusion of buffers poised at different redox potentials or by illumination. The perfusion-induced ATR-FTIR difference spectra for the oxidation of the primary electron donor P in the RCs of the purple bacteria Rb. sphaeroides and Rp. viridis and P700 in the photosystem 1 of Synechocystis 6803, as well as the Q(A)/Q(A) transition of the quinone acceptor (Q(A)) in Rb. sphaeroides RCs are reported for the first time. They are compared with the light-induced ATR FTIR difference spectra P+Q(A)/PQ(A) for the RCs of Rb. sphaeroides and P700+/P700 for photosystem 1. It is shown that the perfusion-induced and light induced ATR-FTIR difference spectra recorded on the same RC film display identical signal to noise ratios when they are measured under comparable conditions. The ATR-FTIR difference spectra are very similar to the equivalent FTIR difference spectra previously recorded upon photochemical or electrochemical excitation of these RCs in the more conventional transmission mode. The ATR-FTIR technique requires a smaller amount of sample compared with transmission FTIR and allows precise control of the aqueous environment of the RC films. PMID- 12083677 TI - Response rate viewed as engagement bouts: resistance to extinction. AB - Rats obtained food pellets by nose poking a lighted key, the illumination of which alternated every 50 s during a session between blinking and steady, signaling either a relatively rich (60 per hour) or relatively lean (15 per hour) rate of reinforcement. During one training condition, all the reinforcers in the presence of the rich-reinforcement signal were response dependent (i.e., a variable-interval schedule); during another condition only 25% were response dependent (i.e., a variable-time schedule operated concurrently with a variable interval schedule). An extinction session followed each training block. For both kinds of training schedule, and consistent with prior results, response rate was more resistant to extinction in the presence of the rich-reinforcement signal than in the presence of the lean-reinforcement signal. Analysis of interresponse time distributions from baseline showed that differential resistance to extinction was not related to baseline differences in the rate of initiating response bouts or in the length of bouts. Also, bout-initiation rate (like response rate) was most resistant to extinction in the presence of the rich reinforcement signal. These results support the proposal of behavioral momentum theory (e.g., Nevin & Grace, 2000) that resistance to extinction in the presence of a discriminative stimulus is determined more by the stimulus-reinforcer (Pavlovian) than by the stimulus-response-reinforcer (operant) contingency. PMID- 12083679 TI - Concurrent schedules: short- and long-term effects of reinforcers. AB - Five pigeons were trained on concurrent variable-interval schedules in a switching-key procedure. The overall rate of reinforcement was constant in all conditions, and the ratios of reinforcers obtainable on the two alternatives were varied over seven levels. Each condition remained in effect for 65 sessions, and the last 50 sessions of data from each condition were analyzed. The most recently obtained reinforcer had the largest effect on current preference, but each of the eight previously obtained reinforcers had a small measurable effect. These effects were larger when the reinforcer ratio was more extreme. A longer term effect of reinforcement was also evident, which changed as a function of the reinforcer ratio arranged. More local analyses showed regularities at a reinforcer-by-reinforcer level and large transient movements in preference toward the just-reinforced alternative immediately following reinforcers, followed by a return to stable levels that were related to the reinforcer ratio in effect. The present data suggest that the variables that control choice have both short- and long-term effects and that the short-term effects increased when the reinforcer ratios arranged were more extreme. PMID- 12083678 TI - Preference and resistance to change with constant- and variable-duration terminal links: independence of reinforcement rate and magnitude. AB - Pigeons responded in a three-component multiple concurrent-chains procedure in which the variable-interval reinforcement schedules were the same across components but magnitudes differed across components. The terminal links were arranged either as a variable delay followed by presentation of a reinforcer ("variable duration") or as a fixed period of access to the schedule during which a variable number of reinforcers could be earned ("constant duration"). Relative reinforcement rate was varied parametrically across both types of conditions. After baseline training in each condition, resistance to change of terminal-link responding was assessed by delivering food during the initial links according to a variable-time schedule. Both preference and resistance to change were more sensitive to reinforcement-rate differences in the constant-duration conditions. Sensitivities of preference and resistance to change to relative reinforcement rate did not change depending on relative reinforcement magnitude. Taken together, these results confirm and extend those of prior studies, and suggest that reinforcement rate and magnitude combine additively to determine preference and resistance to change. A single structural relation linking preference and resistance to change describes all the data from this and several related studies. PMID- 12083680 TI - Preratio pausing: effects of an alternative reinforcer on fixed- and variable ratio responding. AB - Seven rats responding under fixed-ratio or variable-ratio schedules of food reinforcement had continuous access to a drinking tube inserted into the operant chamber. Under different conditions they could drink either tap water or one of two saccharin solutions. In a baseline condition, the drinking bottle was empty. Preratio pausing was observed with both schedules, more so with the fixed-ratio than the variable-ratio schedule, and increasing the concentration of the saccharin solution increased the duration of pausing. Comparisons with baseline performances revealed that the additional pausing was largely, but not entirely, spent drinking. The results support the view that pausing under ratio schedules is a consequence of competition between the scheduled reinforcer and alternative reinforcers that also are available within the experimental environment. PMID- 12083681 TI - Response-initiated imaging of operant behavior using a digital camera. AB - A miniature digital camera, QuickCam Pro 3000, intended for use with video e mail, was modified so that snapshots were triggered by operant behavior emitted in a standard experimental chamber. With only minor modification, the manual shutter button on the camera was replaced with a simple switch closure via an I/O interface controlled by a PC computer. When the operant behavior activated the I/O switch, the camera took a snapshot of the subject's behavior at that moment. To illustrate the use of the camera, a simple experiment was designed to examine stereotypy and variability in topography of operant behavior under continuous reinforcement and extinction in 6 rats using food pellets as reinforcement. When a rat operated an omnidirectional pole suspended from the ceiling, it also took a picture of the topography of its own behavior at that moment. In a single session after shaping of pole movement (if necessary), blocks of continuous reinforcement, in which each response was reinforced, alternated with blocks of extinction (no reinforcement), with each block ending when 20 responses had occurred. The software supplied with the camera automatically stored each image and named image files successively within a session. The software that controlled the experiment also stored quantitative data regarding the operant behavior such as consecutive order, temporal location within the session, and response duration. This paper describes how the two data types--image information and numerical performance characteristics-can be combined for visual analysis. The experiment illustrates in images how response topography changes during shaping of pole movement, how response topography quickly becomes highly stereotyped during continuous reinforcement, and how response variability increases during extinction. The method of storing digital response-initiated snapshots should be useful for a variety of experimental situations that are intended to examine behavior change and topography. PMID- 12083683 TI - Foreword to Schedules of reinforcement. 1957. PMID- 12083682 TI - Schedules of reinforcement with Skinner. 1970. PMID- 12083684 TI - Between the waves: Harvard Pigeon Lab 1955-1960. AB - Six or more graduate students were active in and around the Pigeon Lab in the spring of 1955 and also in 1960, but when I arrived it the fall of 1955 there were none. Looking back in 2001 at that period I can appreciate the unique opportunity for research that I had, as well as the exceptional and productive groups of graduate students who had recently finished their study and research and those who would carry on the tradition of excellence. PMID- 12083685 TI - The watershed years of 1958-1962 in the Harvard Pigeon Lab. AB - During the years 1958-1962, the final years of support by the National Science Foundation for B. F. Skinner's Pigeon Lab in Memorial Hall at Harvard University, 20 or so pigeon experiments (plus some with other organisms) ran concurrently 7 days a week. The research style emphasized experimental analyses, exploratory procedures, and the parametric exploration of variables. This reminiscence describes some features of the laboratory, the context within which it operated, and the activities of some of those who participated in it. PMID- 12083686 TI - The Harvard Pigeon Lab under Herrnstein. AB - The history of the Harvard Pigeon Lab is a history of two periods of remarkable productivity, the first under Skinner's leadership and the second under Herrnstein's. In each period, graduate students flocked to the leader and then began stimulating one another. Chance favored Herrnstein's leadership, too, because an unusually large number of graduate students were admitted in the fall of 1962. In each period, productivity declined as the leader lost interest in the laboratory and withdrew. Directly and indirectly, the laboratory finally died as a result of the cognitive "revolution." Skinner and his students saw the possibility of a natural science of behavior and set about establishing that science based on concepts such as response rate, stimulus control, and schedules of reinforcement. Herrnstein and his students saw that the science could be quantitative and set about making it so, with relative response rate, the matching law, and the psychophysics of choice (analogous to S. S. Stevens' psychophysics). The history might provide a golden research opportunity for someone interested in the impact of such self-organizing research groups on the progress of science. PMID- 12083688 TI - Philosophy of behaviorism. PMID- 12083687 TI - The living legacy of the Harvard Pigeon Lab: quantitative analysis in the wide world. AB - From the Harvard Pigeon Lab of the 1960s arose a behavior-analytic approach that was quantitative and rigorous, rooted in Herrnstein's matching law. Researchers modified the matching law to describe choice behavior in a variety of different settings and examined its relations with other quantitative models. Beginning in the early 1970s, researchers began using the Harvard Pigeon Lab's quantitative framework to study in the laboratory specific aspects of the world outside the laboratory. Much of this work concerned investigations of self-control-choice of a larger, more delayed reinforcer over a smaller, less delayed reinforcer. Experiments using a quantitative framework derived from the matching law have also been conducted outside the laboratory; however, these have been far less frequent. Current and future researchers will benefit the field by devising new, creative ways to investigate the matching law and related quantitative models outside the laboratory. Such research can help to demonstrate the validity of these models as basic principles of behavior, can enhance public opinion of and rewards for such research, and can stimulate further development of the Harvard Pigeon Lab's quantitative approach by using that approach with new variables. PMID- 12083689 TI - Pigeon Lab notable experience. PMID- 12083690 TI - Context matters: my education at the Harvard Pigeon Lab. PMID- 12083692 TI - A view from an outsider. PMID- 12083691 TI - From programmed instruction to pigeons. PMID- 12083694 TI - Reminiscences of a reformed pigeon pusher. PMID- 12083693 TI - The nurturing of a behavior analyst. PMID- 12083695 TI - The Harvard Pigeon Lab, 1970-1998: graduate students and matching law research. PMID- 12083696 TI - The Harvard Pigeon Lab in context. PMID- 12083697 TI - Our Harvard pigeon, rat, dog, and human lab. PMID- 12083698 TI - The matching law illustrates the influence of the Harvard Pigeon Lab. PMID- 12083699 TI - Qualitatively different reinforcers in the Harvard Pigeon Lab. PMID- 12083700 TI - Memories of Memorial Hall. PMID- 12083701 TI - Heavy metal contents in surface soils along the Upper Scheldt river (Belgium) affected by historical upland disposal of dredged materials. AB - For several decades, periodical dredging of river sediments has been necessary to allow for shipping traffic on the river Scheldt. Sediments were disposed along the shores in the alluvial plain without concern for the potential presence of contaminants. The aim of this study was to survey the alluvial plains of the Upper Scheldt river in Belgium for the presence of old dredged sediment landfills, and to appraise the heavy metal contamination at these sites. Up to 82% of the areas that were affected by dredged sediment disposal was found to be polluted by at least one of the metals Cd, Cr, Zn or Pb. Concentrations of Cd, Cr and Zn were, in 10% of the cases, higher than 26, 1900 and 2800 mg/kg, respectively. Cu and Ni concentrations were of no environmental concern on any site. Trends in metal concentrations as a function of location and time were explored and discussed. The highest average concentrations of Cd, Cr, Cu, Pb and Zn were found in the most downstream quarter of the Upper Scheldt. Contents of Pb and Cu were significantly lower for sediments disposed after 1965, but no indication for improvement of the sediment quality with time was observed for Cd, Cr and Zn. The pollution levels encountered warrant for caution as most of the soils affected by historical dredged sediment disposal are currently in use for agriculture, nature development or forestry. PMID- 12083702 TI - Bioaccumulation of As, Cd, Cu, Fe and Pb in wild grasses affected by the Aznalcollar mine spill (SW Spain). AB - The collapse of the tailing dam in the Aznalcollar pyrite mine (SW Spain) occurred in April 1998 and affected approximately 4300 ha along the Agrio and Guadiamar valleys. An urgent soil cleaning up and remediation programme was started just after the accident. Eighteen months later, mineral nutrients and trace elements concentration in soil and two wild grasses--Cynodon dactylon and Sorghum halepense--have been studied. Three types of conditions are distinguished: (a) unaffected soils (control); (b) cleaned up and remediated soils (remediated); and (c) sludge-covered soils left in a fenced plot (non remediated). As, Cd, Cu and Pb in grasses reached toxic levels for the food web in the non-remediated plot, while on remediated soils only Cd reached a toxic level in grass tissues. However, Pb and, to a lesser extent As and Fe, reached also toxic levels in unwashed plants (as they would be ingested by animals) in remediated soils. Both native grasses seem tolerant of trace elements pollution and suitable for stabilisation of spill-affected soils. PMID- 12083703 TI - Myocardial cytochrome c oxidase activity in Swedish moose (Alces alces L.) affected by molybdenosis. AB - Since the mid-1980s, a 'mysterious' wasting disease has been afflicting the moose (Alces alces L.) population of south-western Sweden. In 1994, molybdenosis combined with copper deficiency was suggested as the cause of this complex syndrome of clinical signs, diversity of necropsy findings and changes in trace element concentrations. These findings were corroborated by scientists in many countries by similar observations in other ruminants, particularly cattle and sheep, and also by changes in trace element concentrations and clinical chemical findings in our model experiments with goats. The biochemistry of copper is dependent on a number of copper-dependent enzymes in the animal organism. An important example is cytochrome c oxidase (COX), responsible for oxidative phosphorylation and energy production within the cell. In the present study, COX activity and trace element concentrations were determined in myocardium from affected and healthy moose. Citrate synthase (CS) activity was also measured for use as a mitochondrial marker. COX activity had decreased by 45% and the COX/CS ratio by 37%, while Mo and Na were found to have increased by 140% and 25%, respectively. The increase in Na was indicative of the frequently reported oedematous changes in 'flabby' moose heart. The concentrations of the elements Cu, Mg, Mn, P and Zn had decreased by 20%, 20%, 35%, 7% and 19%, respectively. The simultaneous decrease in COX activity and Cu concentration and the increase in Mo further support the hypothesis that molybdenosis is the cause of the moose disease. PMID- 12083704 TI - Dissolved gaseous mercury formation under UV irradiation of unamended tropical waters from French Guyana. AB - Filtered and non-filtered natural waters from French Guyana were irradiated with lamps emitting within the wavelength range 300-450 nm for 4 days with and without oxygen. Dissolved gaseous mercury (DGM) evolution was observed and quantified in the course of the irradiation. Measurements of total mercury in waters prior to and after the irradiations were also performed. The mass balance in the various mercury species (Hg(total), Hg(reactive) and DGM) proves the capability of the light to extract the mercury linked to the organic matter. DGM evolvement was greater in N2- than in air-saturated solutions, and the formation of volatile oxidized species can account for the inhibiting effect of oxygen. Filtration did not affect the mercury photoreduction, but reduced the formation of DGM in the dark. Great care has to be taken with regard to the following artifact: it was found that DGM originated not only from the natural waters, but also from the experimental device itself when exposed to the light. These non-expected DGM entries were quantitatively evaluated. This stresses the difficulty in measuring mercury at environmental concentrations. PMID- 12083705 TI - Assessment of PAH contamination in estuarine sediments using the equilibrium partitioning-toxic unit approach. AB - Estimates of the potential acute toxicity of PAH residues to aquatic organisms in UK estuarine sediments using the equilibrium partitioning-toxic unit (EqP-TU) approach indicate that sediments from the Clyde and Mersey estuaries and Southampton Water will have the highest mean toxicities. The PAH residues probably originate from a combination of specific industrial sources and inputs from surrounding conurbations via combustion and run-off. Maximum sediment toxicities were evident at specific locations in Southampton Water near a chemical plant outfall and in sediments in docks on the Wear and Clyde. The limitations of the EqP-TU approach, particularly with regard to the potential for the over estimation of toxicity due to high levels of non-bioavailable high molecular weight PAHs occluded in combustion particles are discussed. PMID- 12083706 TI - Investigation of the potential release of polychlorinated dioxins and furans from PCP-treated utility poles. AB - The United States (US) Environmental Protection Agency (EPA) estimated that the use of technical grade pentachlorophenol (PCP) between 1970 and 1995 to treat wood was approximately 400,000 metric tons in the US, and that between 4800 and 36,000 g of 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalents (TEQs) were incorporated annually in treated wood. The EPA has been unable, however, to estimate the rate of release of polychlorinated dibenzo-p-dioxins and dibenzofurans (CDD/Fs) from treated utility poles into the environment. There is some evidence that CDD/Fs leach from treated poles into the surrounding soils, but these studies do not allow for the calculation of a rate of release from this mechanism. Another possible release mechanism is the volatilization of dioxins into the atmosphere, but there are no data to demonstrate, much less quantify, this release. While not directly measuring the release of dioxins from treated utility poles into the environment, this study was designed to examine the potential for such release. The general approach taken was to collect PCP-treated poles of varying ages, to remove and analyze multiple samples from each pole cross-section, and to compare the spatial distribution of CDD/F congeners among poles of different ages. Evidence of concentration-depth profile changes over time may provide insight into the potential for dioxins to migrate through and then out of PCP-treated utility poles. It was found that the CDD/F concentrations were consistently higher in the outer portions of the poles than the center. This trend tends to be most marked in older poles and for the lower chlorinated congeners. The trend for dioxins to concentrate in the outer portions of the pole over time suggest migration within the poles, and this migration may result in some environmental release. Other possible explanations were also offered. PMID- 12083707 TI - A comparative study of manganese and lead levels in human umbilical cords and maternal blood from two urban centers exposed to different gasoline additives. AB - Manganese (Mn) and lead (Pb) are two neurotoxic chemicals and experimental evidence suggests that they can cross the placental barrier. Tetraethyl lead was still in use as an antiknock agent in Paris during the sampling period of the study, while it has been replaced by methylcyclopentadienyl manganese tricarbonyl (MMT) in Canada since 1977. By 1990, MMT was in 100% of gasoline in Canada. In a study of 160 pairs of mothers-neonates in Montreal and 206 pairs in Paris, we compared levels of Mn and Pb in the umbilical cord and in maternal blood. Neonates and mothers had significantly higher Pb levels in Paris where lead additives were still used in gasoline. Geometric mean maternal blood Pb levels were 5.4 microg/dl compared to 2.1 microg/dl in Montreal and cord blood Pb levels were 3.2 microg/dl in Parisian mothers compared to 1.7 microg/dl in Montreal. The prevalence of Paris Pb values superior to the 95th percentile of the Montreal distribution was highly elevated in all media studied. The prevalence of high Mn levels in umbilical cord blood was also significantly higher in Montreal. Surveillance programs are important to limit Pb overexposure and associated neurological effects in neonates where tetraethyl Pb is still in use as a gasoline additive. Since Mn is an essential element and dietary Mn intake may differ between Montreal and Paris, the difference observed with regard to high Mn values between Montreal and Paris cannot, at this time, be attributed to MMT in Montreal's gasoline. Further studies are needed to infer an association between Mn emissions from MMT and prenatal exposure to Mn. PMID- 12083708 TI - Metal contamination at a wood preservation site: characterisation and experimental studies on remediation. AB - The aim of this investigation was to determine the occurrence of As, Cu, Cr and Zn in the soil at an abandoned wood preservation unit and to examine some possible extractants for the contaminants in the soil. The mean As content of the contaminated surface soils (0-10 cm) was 186 mg kg(-1), where as the mean concentrations of Cu, Cr and Zn in soils from the contaminated area were 26, 29 and 91 mg kg(-1), respectively. The elevated As content in the mineral soils is related to adsorption of inorganic As phases in the fine grained fractions, which are characterised by large surface area and high positive surface charge under the current acidic conditions. Cu and Cr were found to be rather mobile, which is reflected in their lower abundance in soils and significant accumulation in sediments in the drainage leaving the area. The fine fraction of the soil (<0.125 mm) has an average metal content increased by nearly 34% as compared to the <2-mm fraction conventionally used for the analysis and assessment of soil contamination. The <2-mm fraction constitutes approximately 65% of the total weight while the fine fraction (<0.125 mm) constitutes approximately 10%. These facts, taken together, are essential for the choice of remediation measures. Oxalate solutions have been tested as extractants for soil remediation. Dark acid oxalate extraction dissolves the amorphous Al- and Fe-oxides and hydroxides and mobilises the adsorbed inorganic As species. Oxalate also acts as a ligand for the cationic heavy metals, releasing them from exchangeable sites. With a three step sequential leaching, up to 98-99% of the metals could be removed. At lower concentrations and higher pH, the leaching decreased to approximately 70%. PMID- 12083709 TI - Towards a global historical emission inventory for selected PCB congeners--a mass balance approach. 1. Global production and consumption. AB - Information on the historical global production and consumption of polychlorinated biphenyls (PCBs) is urgently needed for estimating PCB fluxes to the environment and for interpreting global contamination patterns by these pollutants. This study presents the methodology, principal uncertainties and selected results from an inventory, aiming to quantify the global production and consumption of total PCBs as well as 22 PCB congeners. The available data on the historical production of PCBs and the chemical composition of various technical mixtures have been compiled from the literature. For some producers with less detailed information, the production of individual PCB constituents has been estimated to derive a global estimate for individual homologues and selected congeners. Information on imports, exports and consumption, as well as restrictions on production and imports, has further been compiled for individual countries. These data, along with assumptions on the trade between countries and regions, have been utilised to derive an estimate of the global historical consumption pattern. Although there are substantial uncertainties involved in these estimates, important aspects governing the large scale temporal and spatial patterns are most likely captured in these estimates. In particular, the information on imports and exports for the principal users of PCBs around the time of peak production is considered to be fairly reliable. The estimates account for a reported historical global production of approximately 1.3 million t PCBs, more than 70% of which are tri-, tetra- and pentachlorinated biphenyls. The results further suggest that almost 97% of the global historical use of PCBs have occurred in the Northern Hemisphere. PMID- 12083711 TI - Linear alkylbenzene sulfonates (LAS) in the terrestrial environment. AB - The occurrence of linear alkylbenzene sulfonates (LAS) in a series of soil samples originating from the municipality of Roskilde has been studied. The study includes soil samples from eight different locations with different histories: a preserved natural area that has not been cultured for 50-100 years, a soil that has been ecologically cultured for 40 years, a soil sustainably manured in ecologically culture for 5 years (formerly conventionally cultured) and a soil that has been conventionally cultured using artificial fertilizer. In addition, a soil was studied that had been sludge amended by applying medium amounts of sludge as well as a soil that has been amended with high amounts of sludge for a 25-year period. In the latter case, the sludge amendment was abandoned 6 years before the first sampling, followed by the application of artificial fertilizers. Finally, a meadow in the run-off zone from a sludge storage area was included in the investigations. In addition to the soil samples, selected samples of the applied sludge and other fertilizers were analyzed for their possible content of LAS. Apart from the location where the soil had been heavily sludge-amended and the location situated in the run-off zone of the sludge storage, concentrations of LAS in all soil samples were found to be below approximately 1 mg/kg, which is well below the proposed preliminary soil quality criteria for LAS of 5 mg/kg. On the other hand, the study unambiguously disclosed that in the case of heavy sludge amendment, the proposed soil quality criteria might well be exceeded. PMID- 12083710 TI - Towards a global historical emission inventory for selected PCB congeners--a mass balance approach. 2. Emissions. AB - Accurate and complete data on the global atmospheric releases (emissions) of individual PCB congeners are essential to study source-receptor relationships and contamination patterns in remote areas, such as the Arctic. Information on the anthropogenic emissions may also be useful for the interpretation of measured levels and patterns of atmospheric PCBs. This study builds upon an accompanying paper, presenting an estimate of the global historical production and consumption of 22 PCB congeners (Breivik K, Sweetman A, Pacyna JM, Jones KC. Towards a global historical emission inventory for selected PCB congeners--a mass balance approach. 1. Global production and consumption. Sci Total Environ, submitted). Here, a dynamic mass balance model is elaborated, parameterised and applied in an attempt to estimate the historical anthropogenic emissions as a direct result of the widespread usage of PCBs for a period of 70 years. This paper presents details of the mass balance approach, along with a discussion of the major uncertainties. It is shown that the diversity of historical usage, disposal and accidental release pathways makes it an extremely difficult task to bridge the gap between consumption and emissions, resulting in an equally complex and diverse true emission pattern. These results may therefore at best represent order-of-magnitude estimates only and the results in absolute terms should be treated with great caution. In spite of these uncertainties, useful information can still be gained. The results suggest that temperature is an extremely important factor controlling both emission amount as well as the emission pattern of PCBs. In particular, the emissions of the more chlorinated (and persistent) PCB congeners appear to be significantly influenced by (uncontrolled) high temperature sources. As a consequence, it may prove warranted to undertake further measures to avoid that contaminated materials are subject to elevated temperatures and uncontrolled burning. It is furthermore suggested that efforts should be directed towards a better characterisation and quantification of these potentially important release pathways. In addition, alternative approaches to bridge the gap between consumption and emissions in quantitative terms should be considered in order to improve these estimates. PMID- 12083712 TI - First measurements of carbon tetrachloride and tetrachloroethene in the atmosphere of Athens, Greece. AB - Levels of carbon tetrachloride and tetrachloroethene were determined for the first time in Athens in the summer of 2000. The mean CCl4 and C2Cl4 concentrations were 0.060 and 0.18 ppb, respectively. CCl4 did not exhibit significant variation throughout the measurement period or with wind direction. C2Cl4 under strong eastern winds had less than half the concentrations observed during other meteorological conditions. No diurnal variation was observed for either compound. PMID- 12083713 TI - Campaign launched to eliminate tsetse fly, which has turned much of Africa into a green desert. PMID- 12083714 TI - Copper allergy from dental copper amalgam? AB - A 65-year-old female was investigated due to a gradually increasing greenish colour change of her plastic dental splint, which she used to prevent teeth grinding when sleeping. Furthermore, she had noted a greenish/bluish colour change on the back of her black gloves, which she used to wipe her tears away while walking outdoors. The investigation revealed that the patient had a contact allergy to copper, which is very rare. She had, however, had no occupational exposure to copper. The contact allergy may be caused by long-term exposure of the oral mucosa to copper from copper-rich amalgam fillings, which were frequently used in childhood dentistry up to the 1960s in Sweden. The deposition of a copper-containing coating on the dental splint may be caused by a raised copper intake from drinking water, increasing the copper excretion in saliva, in combination with release of copper due to electrochemical corrosion of dental amalgam. The greenish colour change of the surface of the splint is probably caused by deposition of a mixture of copper compounds, e.g. copper carbonates. Analysis by the X-ray diffraction technique indicates that the dominant component is copper oxide (Cu2O and CuO). The corresponding greenish/bluish discoloration observed on the back of the patient's gloves may be caused by increased copper excretion in tears. PMID- 12083715 TI - Chromium content of selected Greek foods. AB - The total chromium content of a wide variety of Greek foods was determined by graphite furnace atomic absorption spectroscopy (GFAAS). Meat, fish and seafood, cereals and pulses were rich sources of chromium (>0.100 microg/g). Fruits, milk, oils and fats and sugar were poor sources. Differences in chromium content were found between different food classes from Greece and those from some other countries. Based on available food consumption data and chromium levels in this study, it was estimated that the chromium intake of Greeks is 143 microg/day, with vegetables, cereals and meat being the main contributors. PMID- 12083716 TI - Study of the characteristics of CH4 and N2O emission and methods of controlling their emission in the soil-trench wastewater treatment process. AB - In recent years, worldwide concern over global warming has been expressed. It has been reported that domestic wastewater and its treatment processes are sources of CH4 and N2O, designated as greenhouse gases, the reduction of which was noted to be extremely important at the Third Conference of the Framework Convention on Climate Change (Conference Of the Parties; COP3). Here we report a study of a field that has been unexplored until now: analytical evaluation of the properties of the emission of CH4 and N2O and methods of restricting their emission in soil trench wastewater treatment processes, the use of which is spreading, mainly in developing nations. The results have provided the following information. A field fact-finding survey has confirmed that soil trenches emit 9.3-13.9 g CH4 m(-3) and 8.2-12.2 gN2O m(-3) in Japan, and 3.0-4.5 g CH4 m(-3) and 3.3-5.0 g N2O m(-3) in China. The emission properties widely vary according to the structure of the treatment system. The conversion ratio for nitrogen in the wastewater influent to N2O by a soil trench is between 2 and a maximum of 8%, and ranges from a few- to several 10-fold as much as that with the activated sludge method, suggesting that this can be a large source of N2O emission. It has also clearly been shown that the aerobic-anaerobic state inside the treatment system is closely related to its CH4 and N2O emission characteristics. By performing ventilation to maintain the oxidation-reduction potential (ORP) near the trench at an aerobic condition of +200 mV or more, the quantities of CH4 and N2O emitted can be reduced by as much as 50% from the levels without this ventilation, and that this can make a large contribution. PMID- 12083717 TI - The removal of heavy metals from contaminated soil by a combination of sulfidisation and flotation. AB - The possibility of removing cadmium, copper, lead and zinc from Belgian loamy soil by a combination of sulfidisation pre-treatment and Denver flotation was investigated. The potentially available--sulfide convertible--metal content of the metal polluted soil was estimated by EDTA (0.1 M, pH 4.65) extraction and BCR sequential extraction. EDTA extraction is better at approximating the metal percentage that is expected to be convertible into a metal sulfide phase, in contrast to the sequential extraction procedure of 'Int. J. Environ. Anal. Chem. 51 (1993) pp. 135-151' in which transition metals present as iron oxide co precipitates are dissolved by hydroxylammoniumchloride in the second extraction step. To compare the surface characteristics of metal sulfides formed by sulfidisation with those of crystalline metal sulfides, two types of synthetic sediments were prepared and extracted with 0.1 M EDTA (pH 4.65) in anoxic conditions. Separate metal sulfides or co-precipitates with iron sulfide were formed by sulfide conditioning. The Denver flotation of both types of synthetic sediments (kerosene as collector at high background electrolyte concentrations) resulted in similar concentrating factors for freshly formed metal sulfides as for fine-grained crystalline metal sulfides. The selective flotation of metal sulfides after sulfide conditioning of a polluted soil, using kerosene or potassium ethyl xanthate as collectors and MIBC as frother, was studied at high background electrolyte concentrations. The sulfidisations were made in ambient air and inside an anoxic glove box. The concentrating factors corrected by the potentially available metal percentage, determined by 0.1 M EDTA extraction, lie between 2 and 3. The selective flotation of these finely dispersed, amorphous, metal sulfides can possibly be improved by optimising the bubble-particle interaction. PMID- 12083718 TI - Lead poisoning of horses in the vicinity of a battery recycling plant. AB - The diagnosis of lead poisoning in horses living on farmland in the vicinity of a battery recycling plant was based on clinical signs as well as on laboratory findings. Chemical analysis of six surface soils (0-15 cm) and herbage samples taken at different distances to the recycling plant showed very high total lead levels in the closest sites to the facility and a clear decrease with distance. Total lead levels in soil samples ranged from 127 to 5657 mg kg(-1), with more than 70% of lead extractable by EDTA in the most polluted soils. Lead levels in the aerial part of herbage samples were in the range of 113-4741 mg kg(-1). A water washing pre-treatment of the vegetal samples considerably diminished the concentration of lead, suggesting that airborne lead particles from the facility emissions were fixed on the shoots. The analysis of samples taken from six dead horses showed lead concentrations, expressed as mg kg(-1) (d.w.), as follows: blood: 0.20-0.89; liver: 2.5-15; kidney: 1.70-6.75. Lead intake levels, estimated according to the ingestion rate of Grammineae forage, illustrates that the apported lead through the ingestion of vegetation growing in the closest sites to the recycling plant was approximately 99.5 mg Pb/kg body weight/day surpassing the fatal dosage for horses of 2.4 mg Pb/kg body weight/day reported by Hammond and Aronson, Ann NY Acad Sci, 1964; 111: 595-611. PMID- 12083719 TI - Scaling of atmospheric mercury emissions from three naturally enriched areas: Flowery Peak, Nevada; Peavine Peak, Nevada; and Long Valley Caldera, California. AB - With the development of analytical capabilities that allow for almost real time measurement of mercury concentrations in air, the fluxes of mercury between environment compartments is being more carefully scrutinized. Recent advances have demonstrated that the mercury cycle is much more complicated than previously realized. This study quantified the mercury emissions from three areas with low levels of mercury enrichment associated with precious and base metal mineralization and recent volcanic/geothermal activity. Area emissions were calculated using Geographic Information System technology, and in situ derived mercury fluxes and those parameters found to statistically be dominant in controlling emissions. The most important controls on emission strengths were found to be geologic while environmental parameters such as light and temperature were found to drive the diel pattern typically observed for mercury emissions. Calculated area averaged emissions were 18.5, 10.0, and 13.6 ng/m2 h for the Flowery Peak, NV, Peavine Peak, NV, and Long Valley Caldera, CA areas, respectively. These emissions are an order of magnitude higher than values applied in global models for natural sources. This study, along with other recent work, demonstrates that natural sources may contribute more mercury than previously recognized to the atmospheric mercury pool. PMID- 12083720 TI - Topological and developmental gradients of calbindin expression in the chick's inner ear. AB - Mobile intracellular calcium buffers play an important role in regulating calcium flux into mechanosensory hair cells and calbindin D-28k is expressed at high levels in the chick's basilar papilla. We have used RT-PCR, in situ hybridization, and immunohistology to demonstrate that calbindin expression varies systematically according to hair cell position and developmental age. RT PCR using microdissected quarters of the posthatch basilar papilla showed that mRNA levels were lowest in the (low frequency) apex and higher in basal quadrants. In situ hybridization revealed calbindin mRNA in posthatch hair cells and supporting cells, with more intense labeling of hair cells from basal (high frequency) positions. A similar topology was obtained with calbindin antibodies. Neither calbindin riboprobe nor calbindin antibody labeled cochlear neurons. In contrast, a subset of large vestibular neurons and their calyciform endings onto Type I vestibu lar hair cells were strongly labeled by the calbindin antibody, while vestibular hair cells were negative for calbindin immunoreactivity. Likewise, calbindin in situ hybridization was negative for vestibular hair cells but positive in a subset of larger vestibular neurons. Calbindin mRNA was detected in hair cells of the basal half of the papilla at embryonic day 10 (E10) and calbindin immunoreactivity was detected at E12. Hair cells in the apical half of the papilla had equivalent calbindin expression two days later. Immunoreactivity appeared in abneural supporting cells days later than in hair cells, and not until E20 in neurally located supporting cells. These results demonstrate that calbindin message and protein levels are greater in high frequency hair cells. This "tonotopic" gradient may result from the stabilization of a basal-to-apical developmental gradient and could be related at least in part to calcium channel expression along this axis. PMID- 12083721 TI - A multifrequency method for determining cochlear efferent activity. AB - A test based on measures of distortion-product otoacoustic emissions (DPOAEs) was developed in lightly anesthetized guinea pigs and alert rabbits to assess the effective activation or functional "strength" of the cochlear efferent system. The multifrequency method described here used the DP-gram frequency function to evaluate the fast component of the olivocochlear adaptive effect on DPOAE levels over a 2-octave frequency range. An estimate of any concurrent muscle activation was also determined over the identical frequency range by monitoring the levels of the eliciting f1 primary tone throughout its duration. The acoustic reflex, as measured by this f1 level constancy test, did not appear to contribute to the average efferent strength of sedated guinea pigs, but the acoustic reflex did contribute to the average "efferent" strength of awake rabbits. Hence, the average efferent effect in alert rabbits is contaminated by the acoustic reflex, which confounds its interpretation. PMID- 12083722 TI - Relationship between inner-ear fluid pressure and semicircular canal afferent nerve discharge. AB - The present study was designed to determine (1) the transcupular fluid pressure (deltaP) generated across the semicircular canal cupula in response to sinusoidal head rotation, (2) the translabyrinthine dilational pressure (P0) generated across the membranous labyrinth in response to an increase in endolymph fluid volume (hydrops), (3) afferent nerve discharge patterns generated by these distinct pressure stimuli and, (4) threshold values of deltaP and P0 required to elicit afferent neural responses. The experimental model was the oyster toadfish, Opsanus tau. Micromechanical indentation of the horizontal canal (HC) duct and utricular vestibule was used to simulate sinusoidal head rotation and fluid volume injection. Single-unit neural spike trains and endolymph pressure within the ampulla, on both sides of the cupula, were recorded simultaneously. deltaP averaged 0.013 Pa per 1 degrees/s of sinusoidal angular head velocity and P0 averaged 0.2 Pa per 1 nL of endolymph volume injection. The most responsive afferents had a threshold sensitivity to deltaP of 10(-3) Pa and to P0 of 5 x 10( 2) Pa based on a discharge modulation criterion of 1 impulse/s per cycle for 2 Hz pressure stimuli. Neural sensitivity to AP was expected on the basis of transverse cupular and hair bundle deflections. Analysis of mechanics of the end organ, neuronal projections into the crista, and individual neural firing patterns indicates that P0 sensitivity resulted from pressure-induced distension of the ampulla that led to a nonuniform cupular deformation pattern and hair bundle deflections. This explanation is consistent with predictions of a finite element model of the end organ. Results have implications regarding the role of deltaP in angular motion transduction and the role of P0 under transient hydropic conditions. PMID- 12083723 TI - Gene discovery in the auditory system: characterization of additional cochlear expressed sequences. AB - To identify genes involved in hearing, 8494 expressed sequence tags (ESTs) were generated from a human fetal cochlear cDNA library in two distinct sequencing projects. Analysis of the first set of 4304 ESTs revealed clones representing 517 known human genes, 41 mammalian genes not previously detected in human tissues, 487 ESTs from other human tissues, and 541 cochlear-specific ESTs (http://hearing.bwh.harvard.edu). We now report results of a DNA sequence similarity (BLAST) analysis of an additional 4190 cochlear ESTs and a comparison to the first set. Among the 4190 new cochlear ESTs, 959 known human genes were identified; 594 were found only among the new ESTs and 365 were found among ESTs from both sequencing projects. COL1A2 was the most abundant transcript among both sets of ESTs, followed in order by COL3A1, SPARC, EEFY1A1, and TPTI. An additional 22 human homologs of known nonhuman mammalian genes and 1595 clusters of ESTs, of which 333 are cochlear-specific, were identified among the new cochlear ESTs. Map positions were determined for 373 of the new cochlear ESTs and revealed 318 additional loci. Forty-nine of the mapped ESTs are located within the genetic interval of 23 deafness loci. Reanalysis of unassigned ESTs from the prior study revealed 338 additional known human genes. The total number of known human genes identified from 8494 cochlear ESTs is 1449 and is represented by 4040 ESTs. Among the known human genes are 14 deafness-associated genes, including GJB2 (connexin 26) and KVLQT1. The total number of nonhuman mammalian genes identified is 43 and is represented by 58 ESTs. The total number of ESTs without sequence similarity to known genes is 4055. Of these, 778 also do not have sequence similarity to any other ESTs, are categorized into 700 clusters, and may represent genes uniquely or preferentially expressed in the cochlea. Identification of additional known genes, ESTs, and cochlear-specific ESTs provides new candidate genes for both syndromic and nonsyndromic deafness disorders. PMID- 12083724 TI - Gene expression profiles of the rat cochlea, cochlear nucleus, and inferior colliculus. AB - High-throughput DNA microarray technology allows for the assessment of large numbers of genes and can reveal gene expression in a specific region, differential gene expression between regions, as well as changes in gene expression under changing experimental conditions or with a particular disease. The present study used a gene array to profile normal gene expression in the rat whole cochlea, two subregions of the cochlea (modiolar and sensorineural epithelium), and the cochlear nucleus and inferior colliculus of the auditory brainstem. The hippocampus was also assessed as a well-characterized reference tissue. Approximately 40% of the 588 genes on the array showed expression over background. When the criterion for a signal threshold was set conservatively at twice background, the number of genes above the signal threshold ranged from approximately 20% in the cochlea to 30% in the inferior colliculus. While much of the gene expression pattern was expected based on the literature, gene profiles also revealed expression of genes that had not been reported previously. Many genes were expressed in all regions while others were differentially expressed (defined as greater than a twofold difference in expression between regions). A greater number of differentially expressed genes were found when comparing peripheral (cochlear) and central nervous system regions than when comparing the central auditory regions and the hippocampus. Several families of insulin-like growth factor binding proteins, matrix metalloproteinases, and tissue inhibitor of metalloproteinases were among the genes expressed at much higher levels in the cochlea compared with the central nervous system regions. PMID- 12083725 TI - Calpain immunoreactivity and morphological damage in chinchilla inner ears after carboplatin. AB - Carboplatin produces an unusual pattern of damage in the chinchilla inner ear, characterized by early destruction of type I afferent fibers and preferential loss of type I hair cells in the vestibular end organs and inner hair cells (IHCs) in the cochlea. In the present study, we investigated a potential role of calpains, a family of calcium-activated proteases, in carboplatin ototoxicity. Chinchillas received carboplatin (100 mg/kg IP) and were sacrificed 12, 24, 48, or 72 h later for morphological evaluation or immunocytochemistry. Nerve fibers and myelin were the initial sites of increased calpain immunoreactivity (IR) and morphological damage. At 12 h, granular immunoreactive puncta were present within nerve fibers and their myelin sheaths in the spiral ganglion. In the habenula perforata, dense reaction product was present in large vacuoles in the myelin surrounding the nerve fibers. At 24 h, nerve fibers and myelin were destroyed in the habenula, and those in the spiral ganglion showed increased calpain IR and morphological damage. At 72 h, nerve fibers and myelin were completely destroyed. Calpain IR was not a prominent feature of IHCs, type I vestibular hair cells, or ganglion cells at any time after carboplatin. The results show a correlation between calpain IR and carboplatin-induced axon and myelin degeneration. We propose that calpain-induced axonopathy and myelinopathy are primary features of carboplatin ototoxicity in chinchilla. PMID- 12083726 TI - Detection of large interaural delays and its implication for models of binaural interaction. AB - The interaural time difference (ITD) is a major cue to sound localization along the horizontal plane. The maximum natural ITD occurs when a sound source is positioned opposite to one ear. We examined the ability of owls and humans to detect large ITDs in sounds presented through headphones. Stimuli consisted of either broad or narrow bands of Gaussian noise, 100 ms in duration. Using headphones allowed presentation of ITDs that are greater than the maximum natural ITD. Owls were able to discriminate a sound leading to the left ear from one leading to the right ear, for ITDs that are 5 times the maximum natural delay. Neural recordings from optic-tectum neurons, however, show that best ITDs are usually well within the natural range and are never as large as ITDs that are behaviorally discriminable. A model of binaural crosscorrelation with short delay lines is shown to explain behavioral detection of large ITDs. The model uses curved trajectories of a cross-correlation pattern as the basis for detection. These trajectories represent side peaks of neural ITD-tuning curves and successfully predict localization reversals by both owls and human subjects. PMID- 12083727 TI - Conductive hearing loss results in changes in cytochrome oxidase activity in gerbil central auditory system. AB - Conductive hearing loss (CHL) restricts auditory input to an intact peripheral auditory system. Effects of deprivation on the central auditory system (CAS) have been debated, although a number of studies support the hypothesis that CHL can cause modification of CAS structure and function. The present study was designed to test the hypothesis that unilateral CHL results in a decrease in cytochrome oxidase (CO) activity in CAS nuclei that receive major afferent input from the affected ear. Gerbils at postnatal day 12 (P21) or 6-8 weeks underwent left unilateral CHL (malleus removal), cochlear ablation, or a sham surgical procedure. After a survival time of 48 hours or 3 weeks, animals were sacrificed and tissue was processed for cytochrome oxidase histochemistry. Optical density (OD) measurements were made from individual neurons in the anteroventral cochlear nucleus (AVCN) and from medial and lateral dendritic fields in the medial superior olivary nucleus (MSO), the lateral superior olivary nucleus, and the inferior colliculus. The width of the CO-stained neuropil in MSO was also measured as an estimate of dendritic length. OD measures were corrected to neutral areas of the brain. Cochlear ablation caused significant decreases in CO activity in left lower brainstem nuclei, particularly in adult animals. Following CHL, a significant decrease in CO activity was observed in the ipsilateral AVCN and a significant increase was observed in the contralateral AVCN. Cochlear ablation resulted in decreased width of MSO neuropil containing dendrites that receive primary input from the ablated ear. CHL resulted in a significant increase in the width of MSO neuropil on both sides of the brain in the P21 animals that survived 3 weeks but not in P21 animals that survived only 48 hours or in the adult animals. Unilateral CHL is associated with changes in CO activity in the AVCN and may affect MSO dendritic length in younger animals. PMID- 12083728 TI - Symptoms in individuals with small cutaneous veins. AB - The clinical relevance of small cutaneous veins (SCV) is still being discussed. In the Duesseldorf/Essen civil servants study, the prevalence of SCV and the individual symptoms and age-dependent changes were analysed. This cross-sectional study recruited 9935 employees; 9100 could be finally evaluated for this analysis. All volunteers were asked to fill out the questionnaire and were clinically examined. Primarily the clinical findings were documented, adapted to the Basel Study and later modified according to the CEAP classification: (a) class 0 - no visible or palpable clinical signs of venous disease, (b) class 1 - small cutaneous veins, (c) class 1 - reticular veins, (d) class 2 - varicose veins. In all, 64% of the volunteers had no signs of venous disease (class 0: age 41+/-10 years); 10% had small cutaneous veins (class 1: age 44+/-10 years). SCV was more frequent in females (25%) than in males (6%). Only 5% of those with SCV had already consulted a physician. A striking result was that individuals with SCV generally complained about more leg symptoms, of which 'leg swelling' and 'muscle cramps during the night' were the most frequent. 'Continual leg swelling' was reported by 24% of individuals with SCV as opposed to 10% of those without. 'Leg cramps' and 'restless legs' also were more often documented in individuals with SCV (29% vs 22% and 10% vs 7%). These findings were all statistically significant (p<0.001). After adjusting for age and sex, though, there were few or no differences between groups (leg swelling: odds ratio (OR) 1.3; 95% confidence interval (95% CI) 1.1-1.6 and cramps: OR 1.1; 95% CI 0.9-1.3). A gender separate estimation of the rates showed that females suffer more often from any symptom. Regarding 'leg cramps', 'restless legs' and 'itching', the OR were not different for females and males. For 'leg swelling' the age-adjusted OR were significant for women (OR 1.4; 95% Cl 1.1-1.7) compared with men (OR 1.1; 95% Cl 0.7-2). Individuals with SCV seem to have more symptoms compared with healthy people. However, this analysis shows that age and sex are the most relevant explanations for these symptoms. PMID- 12083729 TI - Radiation arteritis following treatment for Wilms' tumor: an unusual case of weight loss. AB - Wilms' tumor is the most common pediatric primary renal cancer and is highly responsive to surgery and chemotherapy. The role of radiotherapy has evolved in the last three decades from the use of stepwise incremental doses in all patients to the current concept of added radiotherapy in advanced cases or in those with unfavorable pathology. To the authors' knowledge, this is the first case of a young woman with prior history of Wilms' tumor and significant weight loss due to radiation arteritis involving the abdominal vasculature. A 31-year-old woman presented with a history of weight loss and severe malnutrition. An angiogram revealed that the aorta was occluded below the renal artery. The celiac artery and the superior mesenteric artery were occluded at the origin, and large intercostal collaterals reconstituted the pelvic circulation. She was initially treated with total parenteral nutrition (TPN) and underwent an infrarenal aortic bypass surgery. Histopathology of the aorta revealed intimal fibrocalcific thickening and changes consistent with radiation-induced arteritis. Later, she was readmitted with progressive hepatic insufficiency. Despite intensive medical therapy, she died of multi-organ failure. PMID- 12083730 TI - Brachial arteriomegaly and true aneurysmal degeneration: case report and literature review. AB - While brachial artery aneurysms are rare and usually of infectious, post traumatic or iatrogenic etiology, true aneurysms of the brachial artery are even more unusual. We report on a large brachial artery aneurysm complicated by chronic contained rupture and partial outflow obstruction. This was observed 19 years after ligation of a radiocephalic (Brescia-Cimino) arteriovenous fistula for hemodialysis that had existed for 2 years. Of 581 brachial artery reconstructions performed at the Cleveland Clinic Foundation between January 1989 and December 2000, only three involved repairs of brachial artery aneurysms; only the reported case was a true aneurysm of degenerative origin for an incidence of 0.17% (1/581). The management of brachial artery aneurysms is described and the pertinent literature was reviewed. PMID- 12083731 TI - Postmethionine-load homocysteine determination for the diagnosis hyperhomocysteinaemia and efficacy of homocysteine lowering treatment regimens. AB - Substantial epidemiological evidence supports the vision that moderate hyperhomocysteinaemia is a graded and independent cardiovascular risk factor. The additional value of the methionine loading test for the assessment of hyperhomocysteinaemia continues to be disputed. This overview presents the historical background for the rationale of the methionine loading test and describes determinants and variability of the postmethionine-load homocysteine concentration. The association of postmethionine-load homocysteine concentrations and the risk of cardiovascular events is discussed. Furthermore, the results of homocysteine lowering treatment on postmethionine-load homocysteine are given. Up to 50% of subjects with hyperhomocysteinaemia can only be detected after performing a methionine loading test; these subjects have a normal fasting homocysteine. Both fasting and postmethionine-load homocysteine concentrations are influenced by serum folate and creatinine concentrations and by the methylenetetrahydrofolate reductase genotype, and may be subject to a wide intra individual variability (approximately 20%). Cross-sectional studies suggest that cardiovascular risk may increase gradually from postmethionine-load homocysteine concentrations above 38 micromol/l. Supplementation with folic acid 0.5 mg daily adequately reduces postmethionine-load homocysteine; addition of pyridoxine appears to have no further homocysteine lowering effect. Prospective studies supporting the clinical significance of the methionine loading test for individual patient counselling are lacking; it seems, therefore, prudent to restrict this test for research purposes. PMID- 12083732 TI - Renal artery revascularization: percutaneous stent placement is the standard of practice. PMID- 12083733 TI - Microalbuminuria: marker of vascular dysfunction, risk factor for cardiovascular disease. AB - Based on the data from large single and multi-center clinical trials, including the Heart Outcomes Prevention Evaluation (HOPE) study, it is clear that the presence of microalbuminuria is a signal from the kidney that cardiovascular risk is increased and that vascular responses are altered. This is exemplified by studies that have demonstrated that the compensatory vasodilation seen following relief from prolonged ischemia or infusion of vasodilators such as nitroglycerin is blunted in people with microalbuminuria. Thus, the presence of between 30 and 299 mg/day of albumin in the urine is associated with abnormal vascular responsiveness, which may be the result of more advanced atherosclerosis and not necessarily related to the presence of hypertension or renal disease. Agents known to reduce the rise in microalbuminuria or actually reduce the level of microalbuminuria, such as ACE inhibitors, angiotensin receptor blockers, HMG-CoA reductase inhibitors, beta blockers, non-dihydropyridine calcium channel blockers and diuretics, have all been shown to reduce cardiovascular mortality and in some cases preserve renal function. This article will present an overview of the data that support the assertion that a reduction in the rise of microalbuminuria is a significant consideration in the selection of agents to treat a given risk factor (cholesterol or blood pressure) to a recommended target goal. Achieving such a goal with agents that also impact microalbuminuria will provide for a more complete cardiovascular risk reduction. PMID- 12083734 TI - Abdominal aortic aneurysms: fresh insights from a novel animal model of the disease. AB - Abdominal aortic aneurysms (AAA) have a high prevalence in aged populations and are responsible for a large number of deaths. Despite the widespread nature of the disease, relatively little is known regarding mechanisms for formation and progression of aortic aneurysms. In part, this lack of knowledge is attributable to a paucity of animal models for this disease. This review summarizes the available animal models of AAA and focuses on a novel model of reproducible AAA generated by infusion of angiotensin II (AngII) into mice rendered hyperlipidemic by the absence of either apolipoprotein E or low-density lipoprotein receptors. AAA generated by AngII infusion have many characteristics of the human disease including marked luminal expansions, perimedial remodeling, inflammation, thrombosis and a link to hyperlipidemia. As in the human disease, male mice are more susceptible to the development of AAA than females. The vascular pathology occurs from the effects of AngII at AT1 receptors present on bone marrow-derived cells. Studies are ongoing to define the mediators responsible for AngII-induced inflammation and degradation of the medial layer of the vascular wall. The AngII induced model of AAA has the potential to provide novel insights into the underlying mechanisms of this disease and assist with the development of pharmacological therapies. PMID- 12083736 TI - Images in vascular medicine. An unusual case of claudication. PMID- 12083735 TI - DNA hypomethylation and methyltransferase expression in atherosclerotic lesions. AB - Arterial smooth muscle cell (SMC) migration and proliferation are central features in atherogenesis. Altered gene expression and cell proliferation in atherosclerotic lesions have some similar characteristics with certain solid tumors and thus might have similar mechanisms that lead to SMC proliferation. Among cancer cells common features are genome-wide hypomethylation which correlates with transformation and tumor progression, and coincident overexpression of methyltransferase (MTase). The purpose of the present study was to analyze whether alterations in DNA methylation and MTase expression are present in atherosclerotic lesions. A significant reduction in genomic 5 methylcytosine content was detected in advanced human atherosclerotic lesions and in lesions of ApoE knock-out mice. SMC were shown to develop hypomethylation in vitro during transformation from a contractile to synthetic phenotype. Balloon denudation of New Zealand White rabbit aorta caused proliferation of intimal SMC with concomitant genomic hypomethylation in the thickened intima. By using in situ hybridization the overall transcriptional activity was found to be increased in clusters of lesion SMC. Marked heterogeneity was seen in MTase mRNA expression in various types of atherosclerotic lesions among intimal and medial SMC. These findings show that (1) genomic hypomethylation occurs during atherogenesis in human, mouse and rabbit lesions and that it correlates with increased transcriptional activity; (2) MTase is expressed in atherosclerotic lesions; and (3) hypomethylation is present in advanced lesions at the same level as in malignant tumors and may affect cellular proliferation and gene expression in atherosclerotic lesions. PMID- 12083737 TI - Images in vascular medicine. Factitial edema. PMID- 12083738 TI - Alteration of gastric ulcerogenic and healing responses in rats with adjuvant induced arthritis. AB - Gastroenteropathy is the most common among patients who use non-steroidal anti inflammatory drugs (NSAIDs) for treatment of inflammatory disorders. It is known that rheumatoid arthritic patients are more susceptible to NSAID-induced gastropathy than other NSAID users. This article reviewed our recent studies concerning the influence of arthritic conditions on gastric ulcerogenic response to NSAID and healing response of chronic gastric ulcers in rats. Gastric lesions induced by indomethacin, one of the conventional NSAIDs, were markedly aggravated in arthritic rats. This increased ulcerogenic response in arthritic rats was attributable to nitric oxide production due to up-regulation of inducible nitric oxide synthase. In arthritic rat stomachs, cyclooxygenase (COX)-2 was also up regulated, where COX-2 selective inhibitors such as rofecoxib or celecoxib provoked gross lesions, although they caused no damage in normal rats. In addition, the healing of chronic gastric ulcers was also delayed in arthritic rats because of less expression of various growth factors such as basic fibroblast growth factors or insulin-like growth factors. Based on these findings, it is concluded that arthritic conditions alter the mucosal ulcerogenic and healing responses in the stomach. Especially, caution should be paid on the use of COX-2 selective inhibitors in rheumatoid arthritic patients. PMID- 12083739 TI - Cardioprotective effects of 9-hydroxyellipticine on ischemia and reperfusion in isolated rat heart. AB - We determined the effect of 9-hydroxyellipticine (9HE) on ryanodine receptor (RyR) and cardiac function after global ischemia in isolated rat hearts. The binding of [3H]-ryanodine in rabbit cardiac sarcoplasmic reticulum was displaced by 9HE in a biphasic manner corresponding to the two sites model with IC50 values of 6.1 microM and 55 mM. The increase of the intracellular Ca2+ concentration induced by caffeine in CHO cells expressing cardiac-type RyR was suppressed by 9HE in a concentration-dependent manner. Pretreatment of the heart with 9HE decreased the total duration of reperfusion-induced ventricular fibrillation (VF) and delayed the onset of VF. There was also a significant recovery of contractile force of ischemic hearts following 9HE. Unlike nifedipine, an L-type Ca2+-channel blocker, 9HE did not suppress the contraction of rat papillary muscles. Thus, 9HE exerts the cardioprotective effects against ischemia /reperfusion injury without changing hemodynamic indices. PMID- 12083740 TI - Endothelium-dependent vasorelaxation induced by black currant concentrate in rat thoracic aorta. AB - We investigated the effect of black currant (BC) concentrate on smooth muscle in rat thoracic aorta. BC concentrate dose-dependently relaxed the norepinephrine (0.1 microM)-precontracted aorta, and the response was abolished after endothelium removal. Both oxyhemoglobin (1 microM), a nitric oxide (NO) scavenger, and IH-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ, 0.5 microM), an inhibitor of guanylyl cyclase (GC), inhibited the relaxing effect of BC concentrate. NG-nitro-L-arginine methyl ester (L-NAME, 10 microM), a nitric oxide synthase (NOS) inhibitor, inhibited the relaxation, and the subsequent addition of L-arginine (1 mM), a NOS substrate, reversed the inhibitory effects of L-NAME. Neither indomethacin (10 microM), an inhibitor of cyclooxygenase, nor atropine (1 microM), an antagonist of muscarinic receptors, modified the effect of BC concentrate. Diphenhydramine (3 microM) and chlorpheniramine (2 microM), selective antagonists of H1-receptors, inhibited the relaxation, but cimetidine (0.3 mM), a selective antagonist of H2-receptors, did not affect the relaxation. These results indicate that, in the rat aorta, BC concentrate enhances synthesis of NO, which subsequently induces the endothelium-dependent vasorelaxation via the H1-receptors on the endothelium. PMID- 12083741 TI - Beta-hydroxybutyrate, a cerebral function improving agent, protects rat brain against ischemic damage caused by permanent and transient focal cerebral ischemia. AB - In our previous study, beta-hydroxybutyrate (BHB) was found to prolong survival time and to inhibit cerebral edema by improving energy metabolism in the hypoxia, anoxia and global cerebral ischemia models. In this study, the cerebroprotective effect of BHB was examined in rats with permanent (p)-occlusion and transient (t) occlusion of middle cerebral artery (MCA). BHB (30 mg x kg(-1) x h(-1) was continuously administered through the femoral vein. In rats with p-MCA occlusion, BHB significantly reduced infarct area at 24 h after the occlusion, but not at 72 h after the occlusion. In rats with 2-h t-MCA occlusion followed by 22-h reperfusion, BHB significantly reduced cerebral infarct area, edema formation, lipid peroxidation and neurological deficits. Moreover, in the t-MCA occlusion model, delayed administration of BHB started at 1 h after the initiation of the MCA occlusion also significantly reduced cerebral infarct area. Taking together the results obtained in our previous study into account, these results indicate that BHB decreased cerebral edema formation and infarct area by improving of the cerebral energy metabolism during ischemia and by inhibition of lipid peroxidation after reperfusion. PMID- 12083742 TI - Existence of ionotropic glutamate receptor subtypes in cultured rat retinal ganglion cells obtained by the magnetic cell sorter method and inhibitory effects of 20-hydroxyecdysone, a neurosteroid, on the glutamate response. AB - Glutamate and neurosteroids are known to exist in retinal ganglion cells (RGC). Therefore, patch clamp studies using the whole-cell recording method were performed to determine whether or not ionotropic glutamate receptor subtypes, i.e., N-methyl-D-aspartate (NMDA), alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) and kainate receptors, were present on RGC obtained by the magnetic cell sorter (MACS) method and cultures. In addition, the effects of 20 hydroxyecdysone (20-HE), a neurosteroid, on inward currents induced by NMDA, AMPA and kainate were examined at a holding potential of -60 mV. The current-voltage relationship for NMDA in the presence of glycine and Mg2+-free, as well as those for AMPA and kainate were linear, with a reversal potential of around 0 mV. NMDA induced currents were blocked by MK-801, while both AMPA- and kainate-induced currents were blocked by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). Application of 20-HE in the bath resulted in significant inhibitions on NMDA-, AMPA- and kainate-induced currents. Thus, NMDA, AMPA and kainate receptors were confirmed to exist on MACS-separated cultured RGC. Moreover, 20-HE inhibited NMDA receptor mediated currents most prominently and AMPA- and kainate-mediated currents moderately, suggesting that neurosteroids may be playing a role in modulating glutamate-mediated transmission in RGC, and 20-HE might be useful for preventing glutamate neurotoxicity. PMID- 12083743 TI - Mechanisms underlying the activation of large conductance Ca2+-activated K+ channels by nordihydroguaiaretic acid. AB - The mechanisms underlying the activation of large conductance Ca2+-activated K+ (BK) channel by nordihydroguaiaretic acid (NDGA) were examined in human embryonic kidney (HEK293) cells, where BK channel alpha (BKalpha) or a plus beta1 subunit (BKalphabeta1) was heterologously expressed, and also in freshly isolated porcine coronary arterial smooth muscle cells (PCASMCs). The activity of both BKalpha and BKalphabeta1 channels was increased by 10 microM NDGA in similar manners, indicating the selective action on the a subunit to increase Ca2+ sensitivity. The application of NDGA to PCASMCs induced outward current and hyperpolarization under voltage and current clamp, respectively, in a concentration-dependent manner (> or = 3 microM). These effects were blocked by 100 nM iberiotoxin. Electrical events induced by NDGA (> or = 10 microM) were, unexpectedly, associated with the increase in [Ca2+]i. After the treatment with caffeine and ryanodine, the [Ca2+]i increase by NDGA was markedly reduced and the hyperpolarization by NDGA was attenuated. The Ca2+ release by 10 microM NDGA was preceded by membrane depolarization of mitochondria. These results indicate that BK channel opening by NDGA in PCASMCs is due to the direct action on a subunit and also to Ca2+ release from sarcoplasmic reticulum, presumably via, at least in part, the inhibition of mitochondria respiration. PMID- 12083744 TI - The nitric oxide donor NOC12 protects cultured astrocytes against apoptosis via a cGMP-dependent mechanism. AB - We examined the effect of 3-ethyl-3-(ethylaminoethyl)-1-hydroxy-2-oxo-1-triazene (NOC12), a nitric oxide (NO) donor, on apoptosis in cultured astrocytes. Reperfusion after hydrogen peroxide (H2O2) exposure caused a decrease in cell viability, loss of mitochondrial membrane potential, caspase-3 activation, DNA ladder formation, and nuclear condensation. NOC12 at 10-100 microM significantly attenuated these apoptotic changes, while the NO donor at 1 mM caused cell injury and exacerbated the H202-induced cell injury. NOC12 increased intracellular cGMP levels in a dose dependent manner with the maximal effect at 100 microM. The protective effect of NOC12 was mimicked by the NO-independent guanylate cyclase activator 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole, and was attenuated by the guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) and the cGMP-dependent protein kinase inhibitor KT5823. ODQ and KT5823 did not block but rather exacerbated the cytotoxic effect of NOC12 at 1 mM. These findings demonstrate that lower concentrations of NOC12 inhibit the H2O2-induced apoptosis of astrocytes in a cGMP-dependent way, but higher concentrations of NOC12 show a toxic effect on astrocytes in a cGMP-independent way. PMID- 12083745 TI - Research and development of donepezil hydrochloride, a new type of acetylcholinesterase inhibitor. AB - A wide range of evidence shows that cholinesterase (ChE) inhibitors can interfere with the progression of Alzheimer's disease (AD). The earliest known ChE inhibitors, namely, physostigmine and tacrine, showed modest improvement in the cognitive function of AD patients. However, clinical studies show that physostigmine has poor oral activity, brain penetration and pharmacokinetic parameters, while tacrine has hepatotoxic liability. Studies were then focused on finding a new type of acetylcholinesterase (AChE) inhibitor that would overcome the disadvantages of these two compounds. During the study, by chance we found a seed compound. We then conducted a structure-activity relationship study of this compound. After four years of exploratory research, we found donepezil hydrochloride (donepezil). Donepezil showed several positive characteristics including the following: 1) It has a novel structure compared to other conventional ChE inhibitors; 2) It shows strong anti-AChE activity and has long lasting efficacy; 3) The inhibitory characteristic of donepezil shows that it is highly selective for AChE as compared to butyrylcholinesterase (BuChE) and showed reversibility; 4) The results of clinical studies on donepezil show a very high significant difference on ADAS cog and CIBIC plus scores of AD patients. Donepezil is currently marketed in 56 countries all over the world. PMID- 12083746 TI - Regulation of guanylate cyclase by ATP and dithiothreitol in rat lung membrane: involvement of an insensitive and a sensitive state to ATP/dithiothreitol stimulation. AB - ATP/dithiothreitol (DTT)-stimulated guanylate cyclase (GC) in lung membrane was stimulated 18-fold by ATP and DTT, and both its activity and atrial natriuretic peptide (ANP)-stimulated GC activity were observed to be additive. ATP/DTT stimulated GC was solubilized by octyl glucoside (OG) to examine the mechanism of ATP/DTT-stimulation. GC in OG-extracts was stimulated maximally 2.5-fold by both ATP, ATPgammaS or AMPPNP, and DTT. Preincubation of OG-extracts at 10 degrees C with AMPPNP and DTT (1st-preincubation) converted GC to an insensitive state to stimulation by both ATP and DTT, and this conversion was partly inhibited by a protein phosphatase-1 inhibitor (10-1,000 nM okadaic acid). On the other hand, ANP-stimulated GC was not converted to an insensitive state to ANP/ATP stimulation by the 1st-preincubation. Subsequent preincubation of OG-extracts at 10 degrees C with both DTT and, ATP or ATPgammaS but not AMPPNP converted GC to a state sensitive to ATP/DTT-stimulation, and this conversion was partly inhibited by inhibitors of Ca2+/calmodulin-dependent protein kinase II (KN-62 and KN-93). In contrast, the preincubation with KN-62 and KN-93 had no effect on ANP stimulated GC activity. The results suggested that phosphorylation was involved in the regulation of ATP/DTT-stimulated GC sensitivity to ATP/DTT-stimulation and that ATP/DTT-stimulated GC activity was likely to be a different type from ANP stimulated GC activity. PMID- 12083747 TI - A nuclear factor-kappaB inhibitor BAY 11-7082 suppresses endothelin-1 production in cultured vascular endothelial cells. AB - BAY 11-7082, an inhibitor of nuclear factor-kappaB (NF-kappaB), which prevents a step of the phosphorylation of inhibitory protein IkappaB bound to NF-kappaB, suppressed basal and tumor necrosis factor (TNF)-alpha-induced prepro endothelin (ET)-1 mRNA expression and NF-kappaB activation in cultured vascular endothelial cells. BAY 11-7082 significantly decreased basal and TNF-alpha-induced ET-1 release from endothelial cells. These results indicate that the inhibition of NF kappaB activation contributes to the suppressive effect of BAY 11-7082 on ET-1 gene expression and ET-1 release, thereby suggesting that NF-kappaB plays an important role in the regulation of ET-1 production. PMID- 12083748 TI - Both metabotropic glutamate I and II receptors mediate augmentation of dopamine release from the striatum in methamphetamine-sensitized rats. AB - The role of metabotropic glutamate receptor (mGluR) on dopamine overflow from the striatum was studied in methamphetamine (MAP)-sensitized rats. The increase of dopamine release by MAP was significantly inhibited by perfusion of a mGluR antagonist R,S-alpha-methyl-4-carboxyphenylglycine. The perfused mGluR agonist [S,3R-1-aminocyclopentane-1,3-dicarboxylic acid enhanced the dopamine level. The enhancement was significantly attenuated by co-perfusion of a mGluR group I antagonist (S)-4-carboxy-3-hydroxyphenylglycine or a mGluR group II antagonist R,S-a-methyl-4-tetrazolylphenylglycine. These suggest that both mGluR group I and II mediate augmentation of dopamine release in MAP-sensitized rats. PMID- 12083750 TI - Another helpful article--beliefs. PMID- 12083749 TI - Modulation by 5-hT2A receptors of aggressive behavior in isolated mice. AB - The present study examines whether isolation-rearing affects sensitivity of 5 hydroxytryptamine (5-HT)2A receptors and the functional interaction between 5 HTIA and 5-HT2A receptors in mice. The 5-HT2A-receptor agonist (+/-)-1-(2,5 dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride (DOI)-induced head twitch response was significantly greater in isolated mice than in grouped mice. DOI increased isolation-induced aggressive behavior, and the 5-HT2A-receptor antagonist ritanserin decreased it. The 5-HTIA-receptor agonist (S)-5-[3-[(1,4 benzodioxan-2-ylmethyl)amino]propoxy]-1,3-benzodioxole HCl (MKC-242) inhibited the DOI-enhanced aggressive behavior. MKC-242 inhibited DOI-induced head twitch response. These findings suggest that 5-HT2A receptors play a role in aggressive behavior in isolated mice and imply that the antiaggressive effect of MKC-242 may be mediated partly by the inhibition of 5-HT2A-receptor function. PMID- 12083751 TI - Success in decreasing restraint and seclusion. PMID- 12083752 TI - Helping children respond to 9/11. PMID- 12083753 TI - Preliminary effects of brief school-based prevention approaches for reducing youth suicide--risk behaviors, depression, and drug involvement. AB - PROBLEM: Few empirically tested, school-based, suicide-prevention programs exist. The purpose of this study was to evaluate the postintervention efficacy of Counselors-CARE (C-CAST) and Coping and Support Training (CAST) vs. "usual care" controls for reducing suicide risk. METHODS: A randomized prevention trial; 341 potential dropouts, 14 to 19 years old, from seven high schools (52% female, 56% minorities) participated. Trend analyses using data from three time points assessed over time changes. FINDINGS: Significant decreases occurred for all youth in suicide-risk behaviors, depression, and drug involvement. Intervention specific effects occurred for decreases in depression. CONCLUSIONS: School-based prevention approaches are feasible and show promise for reducing suicidal behaviors and related depression. PMID- 12083754 TI - Caring for the involuntarily hospitalized adolescent: the issue of power in the nurse-patient relationship. AB - TOPIC: The power relations between involuntarily hospitalized adolescents and their caregivers from the viewpoint of theory development in nursing and the concept of medicalization. PURPOSE: To contribute to the development of nursing theories that can guide nursing care of involuntarily hospitalized adolescents by examining the power relations between the adolescents and their caregivers. SOURCES: Published literature in medical sociology, and nursing and psychiatric ethics. Nursing and psychiatric literature, in which Michel Foucault's theories are used to explicate the disciplinary power of psychiatry. CONCLUSIONS: To empower involuntarily hospitalized adolescents nurses must both recognize the inevitable power imbalance and seek guidance from the egalitarian ideals of nursing. Nurses can draw from postmodern thinkers who dismiss the idea that one theory can explain human relationships. PMID- 12083755 TI - Social support in pregnant and parenting adolescents: research, critique, and recommendations. AB - ISSUE AND PURPOSE: Adolescent pregnancies continue to occur at an alarming rate in the United States, resulting in adverse outcomes for both the adolescent and her baby. Since social support has been shown to improve pregnancy and parenting outcomes, a critique of research in this area is presented. SOURCES: Published literature. CONCLUSIONS: Multisite research studies, which are longitudinal in design, are needed, with particular attention paid to variations in the need for social support by stage of adolescence. PMID- 12083756 TI - Eye on Washington: focusing on nursing numbers. PMID- 12083757 TI - Overexpression and biochemical characterization of beta-1,3-N acetylgalactosaminyltransferase LgtD from Haemophilus influenzae strain Rd. AB - The lipopolysaccharide of capsule deficient Haemophilus influenzae strain Rd contains an N-acetylgalactosamine residue attached to the terminal globotriose moiety in the Hex5 glycoform. Genome analysis identified an open reading frame HI1578, referred to as lgtD, whose amino acid sequence shows significant level of similarity to a number of bacterial glycosyltransferases involved in lipopolysaccharide biosynthesis. To investigate its function, overexpression and biochemical characterization were performed. Most of the protein was obtained in a highly soluble and active form. By using standard glycosyltransferase assay and HPLC, we show that LgtD is an N-acetylgalactosaminyltransferase with high donor substrate specificity and globotriose is a highly preferred acceptor substrate for the enzyme. The K(m) for UDP-GalNAc and globotriose are 58 microM and 8.6 mM, respectively. The amino acid sequence of the enzyme shows the conserved features of family II glycosyltransferases. This is the first N acetylgalactosaminyltransferase identified from H. influenzae, which shows potential application in large-scale synthesis of globo-series oligosaccharides. PMID- 12083758 TI - The antioxidant activity of phloretin: the disclosure of a new antioxidant pharmacophore in flavonoids. AB - Phloretin is a dihydrochalcone flavonoid that displays a potent antioxidant activity in peroxynitrite scavenging and the inhibition of lipid peroxidation. Comparison with structurally related compounds revealed that the antioxidant pharmacophore of phloretin is 2,6-dihydroxyacetophenone. The potent activity of 2,6-dihydroxyacetophenone is due to stabilisation of its radical via tautomerisation. The antioxidant pharmacophore in the dihydrochalcone phloretin, i.e., the 2,6-dihydroxyacetophenone group, is different from the antioxidant pharmacophores previously reported in flavonoids. PMID- 12083759 TI - Sensitivity enhancement of DNA sensors by nanogold surface modification. AB - A novel amplified microgravimetric gene sensing system was developed using quartz crystal microbalance modified by gold nanoparticles anchored on its 1,6 hexanedithiol modified gold electrode surface, and ultrasensitive detection of DNA hybridization was accomplished at the level of at least 2 x 10(-16) M. PMID- 12083760 TI - Significant correlation of the SCN1A mutations and severe myoclonic epilepsy in infancy. AB - To investigate the possible correlation between genotype and phenotype of epilepsy, we analyzed the voltage-gated sodium channel alpha1-subunit (SCN1A) gene, beta1-subunit (SCN1B) gene, and gamma-aminobutyric acid(A) receptor gamma2 subunit (GABRG2) gene in DNAs from peripheral blood cells of 29 patients with severe myoclonic epilepsy in infancy (SME) and 11 patients with other types of epilepsy. Mutations of the SCN1A gene were detected in 24 of the 29 patients (82.7%) with SME, although none with other types of epilepsy. The mutations included deletion, insertion, missense, and nonsense mutations. We could not find any mutations of the SCN1B and GABRG2 genes in all patients. Our data suggested that the SCN1A mutations were significantly correlated with SME (p<.0001). As we could not find SCN1A mutations in their parents, one of critical causes of SME may be de novo mutation of the SCN1A gene occurred in the course of meiosis in the parents. PMID- 12083762 TI - Selection of human antibody fragments on the basis of stabilization of the variable domain in the presence of target antigens. AB - Here we report a novel method for selecting human antibody fragments from nonimmunized variable domain libraries. The antibody fragments are selected on the basis of stabilization of the variable domain fragment (F(v)) in the presence of target antigens ("open sandwich selection"). One variable domain is displayed on phages and another is prepared as soluble molecules. These two reagents are mixed with the biotinylated target molecule and ternary complexes are captured by using streptavidin-conjugated magnet beads. After extensive washing, enriched clones are eluted by using target antigen. Some of the clones selected after 3 rounds are prepared as soluble domains, which then undergo another selection process. We obtained several human antibody fragments specific for human soluble erythropoietin receptor by using this method. Our method minimizes several of the disadvantages associated with human antibody selection through a phage-display system, such as construction of a large-scale library, deletion of genes during selection, and nonspecific binding. PMID- 12083761 TI - Insights into the molecular mechanism of apoptosis induced by TNF-alpha in mouse epidermal JB6-derived RT-101 cells. AB - The mammalian response to stress is complex, often involving multiple signaling pathways that act in concert to influence cell fate. To examine potential interaction between the signaling cascade, we have focused on the effects of a model apoptotic system in a single cell type sensitive to TNF-alpha induced apoptosis through an examination of the relative influences of MAPKs as well as transcription factors AP-1, NF-kappaB, and various survival genes in determining apoptosis. Our results show that ERKs decreased transiently or remain unchanged, JNK decreased robustly, whereas c-Jun increased transiently, thereby indicating that members of MAPK family are differentially regulated in response to TNF-alpha induced apoptosis, whereas NF-kappaB protein expression decreased transiently and activity decreased at 24 h post-treatment. The survival genes Bcl-2, Bcl-XL, and survivin act independently and downstream of ERK and JNK to decrease the survival of TNF-alpha treated RT-101 cells. The results also suggest the involvement of the mitochondria and cytochrome c. Caspase-3 appears to be a part of a downstream event. PMID- 12083763 TI - Apoptosis is an adaptive response in bovine preimplantation embryos that facilitates survival after heat shock. AB - Heat shock compromises development of preimplantation bovine embryos and the percentage of blastomeres labeled as TUNEL-positive. It was hypothesized that TUNEL labeling represents apoptosis and that apoptosis after heat shock is beneficial for continued embryonic development. To test these hypotheses, experiments were performed with z-DEVD-fmk, an inhibitor of group II caspases, on heat shock responses of embryos > or =16-cell stage at day 4 after insemination. Heat shock of 41 degrees C for 9 h increased group II caspase activity and the proportion of TUNEL positive cells; z-DEVD-fmk blocked these effects. The reduction in development of embryos exposed to heat shock for 6-9 h was magnified in the presence of z-DEVD-fmk. Results indicate that group II caspases mediate heat-induced apoptosis in bovine embryos and that inhibition of these caspases has a detrimental effect on embryonic resistance to heat shock. Apoptosis can be viewed as an adaptative mechanism to allow embryonic survival and development following stress. PMID- 12083764 TI - Transcription of Xanthomonas campestris prt1 gene encoding protease 1 increases during stationary phase and requires global transcription factor Clp. AB - Xanthomonas campestris pv. campestris produces three proteases, Prt1, Prt2, and Prt3, the first two of which are involved in pathogenicity. In this study, nucleotide A 84 nt upstream of the prt1 start codon, which is 8 nt downstream of the -10 sequence, was determined as the transcription start site by the 5(') RACE (rapid amplification of cDNA ends) method. Using Pprt1-lacZ transcriptional fusion constructs for assays, several interesting characteristics of prt1 promoter were revealed. The expression is inducible by LB medium or casein proteins and involves the global transcription factor Clp (cyclic AMP receptor protein-like protein). The region containing bp -392 to -80 relative to the prt1 translation initiation codon is required for maximal expression, in which bp -392 to -207 responds to the Clp-mediated regulation and the induction. In presence of inducers and the clp wild-type background, the levels of expression continue to increase following cell growth until 30 h after the cultures entering stationary phase. Since prt1 promoter shows no response to stressful conditions and neither growth nor cell viability is affected by prt1 mutation, Prt1 appears to be a secondary metabolite of X. campestris pv. campestris. PMID- 12083765 TI - Characterization and daily variation of nitrate reductase in Gracilaria tenuistipitata (Rhodophyta). AB - A daily rhythm in the activity of nitrate reductase (NR: EC 1.6.6.1) isolated from the marine red algae Gracilaria tenuistipitata is shown to be attributable to changes in amounts of the protein. The enzyme was purified in four steps: ion exchange Q-Sepharose separation, ammonium sulfate precipitation, gel filtration on Sephacryl S-300, and affinity chromatography on Affigel-blue resin. This purification procedure yielded an active purified NR of about 500-fold with a recovery of 85%. The SDS-PAGE silver staining of purified NR revealed a 110 kDa single band. Non-denaturated protein showed a molecular mass of 440 kDa on gel filtration comparing with SDS-PAGE, the enzyme is apparently composed of four identical subunits. In extracts of algae grown under either constant dim light or a light-dark cycle, the activity of NR exhibited a daily rhythm, peaking at midday phase as does photosynthesis. Staining with monoclonal antibodies, raised against NR from Porphyra yezoensis, showed that the amount of protein changes by a factor of about 12, with a maximum occurring in the midday phase. PMID- 12083767 TI - Curcumin (diferuloylmethane), a singlet oxygen ((1)O(2)) quencher. AB - Curcumin (diferuloylmethane) is a major component of food flavoring turmeric (Curcuma longa), and has been reported to be anticarcinogenic and anti inflammatory. Although curcumin was shown to have antioxidant properties, its exact antioxidant nature has not been fully investigated. In this report we have investigated the possible antioxidant properties of curcumin using EPR spectroscopic techniques. Curcumin was found to inhibit the (1)O(2)-dependent 2,2,6,6-tetramethylpiperidine N-oxyl (TEMPO) formation in a dose-dependent manner. (1)O(2) was produced in a photosensitizing system using rose bengal as sensitizer, and was detected as TEMP-(1)O(2) adducts by electron paramagnetic resonance (EPR) spectroscopic techniques using TEMP as a spin-trap. Curcumin at 2.75 microM caused 50% inhibition of TEMP-(1)O(2) adduct formation. However, curcumin only marginally inhibited (24% maximum at 80 microM) reduction of ferricytochrome c in a xanthine-xanthine oxidase system demonstrating that it is not an effective superoxide radical scavenger. Additionally, there was minor inhibition of DMPO-OH adduct formation by curcumin (solubilized in ethanol) when an ethanol control was included in the EPR spin-trapping study, suggesting that curcumin may not be an effective hydroxyl radical scavenger. Together these data demonstrate that curcumin is able only to effectively quench singlet oxygen at very low concentration in aqueous systems. PMID- 12083766 TI - C-terminus modification of Streptomyces clavuligerus deacetoxycephalosporin C synthase improves catalysis with an expanded substrate specificity. AB - The biosynthesis of cephalosporins is catalyzed by deacetoxycephalosporin C synthase (DAOCS). Based on computational, biochemical, and structural analyses, it has been proposed that modification of the C-terminus of DAOCS might be a constructive strategy for engineering improvement in enzyme activity. Therefore, five hydrophilic residues namely N301, Y302, N304, R306, and R307 located in proximity to the C-terminus of Streptomyces clavuligerus DAOCS (scDAOCS) were selected and each substituted with a hydrophobic leucine residue. Substitutions at positions 304, 306, and 307 created mutant scDAOCSs with improved efficiencies in penicillin analog conversion up to 397%. And since it has been previously advocated that the C-terminus is crucial for guiding substrate entry, a truncated mutant DAOCS was constructed to assess its involvement. The truncation of the C terminus at position 310 in the wild-type scDAOCS resulted in reduction of indiscriminate conversion of penicillin analog but this defect was compensated by the replacement of asparagine with leucine at position 304. PMID- 12083768 TI - The DNA-binding activity of protein disulfide isomerase ERp57 is associated with the a(') domain. AB - ERp57 belongs to the protein disulfide isomerases, a family of homologous proteins mainly localized in the endoplasmic reticulum and characterized by the presence of a thioredoxin-like folding domain. ERp57 is a protein chaperone with thiol-dependent protein disulfide isomerase and additional activities and recently it has been shown to be involved, in cooperation with calnexin or with calreticulin, in the correct folding of glycoproteins. However, we have demonstrated that the same protein is also present in the nucleus, mainly associated with the internal nuclear matrix fraction. In vitro studies have shown that ERp57 has DNA-binding properties which are strongly dependent on its redox state, the oxidized form being the competent one. A comparison study on a recombinant form of ERp57 and several deletion mutants, obtained as fusion proteins and expressed in Escherichia coli, allowed us to identify the C-terminal a(') domain as directly involved in the DNA-binding activity of ERp57. PMID- 12083769 TI - SREBP-2 and NF-Y are involved in the transcriptional regulation of squalene epoxidase. AB - The expression of squalene epoxidase (SE) is highly regulated transcriptionally by cholesterol. To elucidate these molecular mechanisms, we isolated the human and rat genomic clones. The entire human SE gene was about 24 kb long and organized into 11 exons with 10 introns. Unidirectional deletion analysis of the human 5(')-flanking region indicated that the sequence between -264 and -230 bp conferred cholesterol sensitivity on a reporter gene. This region contained a potential copy of consensus sterol regulatory element (SRE) sequence (CCACGCAAC) previously identified in the promoter of cholesterogenic and its related genes. The transcriptional activation observed under overexpression of sterol regulatory element binding protein-2 (SREBP-2) supported the functional role of the SRE sequence. Another deletion analysis showed that the sequence -207 to -192 bp was also active and it contained nuclear factor Y (NF-Y) binding site. Both sites might play critical roles in sterol mediated regulation of SE gene. PMID- 12083770 TI - Processing of the Hepatitis C virus precursor protein expressed in the methylotrophic yeast Pichia pastoris. AB - The expression and processing of the Hepatitis C virus core protein (HCcAg) were analyzed in the methylotrophic yeast Pichia pastoris. Two proteins with 21 (p21) and 23 kDa (p23) were detected in immunoblot with a serum from a chronic carrier patient, as the major products for HCcAg. Both proteins, p21 and p23, produced by proteolytic processing in P. partoris, share the same N-terminal region and reacted with a monoclonal antibody towards the first 35 amino acids of HCcAg. The proteolytic processing of the recombinant polypeptide, having the HCcAg and the first 148 aa of E1 protein, was also confirmed by immunoblot analysis using mAbs with HCcAg and E1 specificities, respectively. The 32 kDa glycosilated E1 protein was then immuno-identified, as well as the processed HCcAg. These data demonstrated the usefulness of P. pastoris, as expression system, to study the processing of HCV structural proteins. PMID- 12083771 TI - Molecular cloning and characterization of dullard: a novel gene required for neural development. AB - In a screen for genes expressed in neural tissues and pronephroi, we isolated a novel gene, named dullard. Dullard protein contains the C-terminal conserved domain of NLI-IF (Nuclear LIM Interactor-Interacting Factor), a protein whose function is not yet characterized. Dullard mRNA was maternally derived and localized to the animal hemisphere. At neurula stages, the expression was in neural regions and subsequently localized to neural tissues, branchial arches, and pronephroi. Using antisense morpholino oligonucleotide-mediated inhibition, we showed that dullard was required for neural development. The translational knock-down of dullard resulted in failure of neural tube development and the embryos consequently showed a reduction of head development. Expression of neural marker genes in dullard-inhibited embryos was also suppressed. These results suggest that dullard is necessary for neural development. PMID- 12083772 TI - Extensive overproduction of the AdhE protein by rng mutations depends on mutations in the cra gene or in the Cra-box of the adhE promoter. AB - Escherichia coli RNase G encoded by the rng gene is involved in degradation of adhE mRNA. Overproduction of the AdhE protein by rng mutants was found to depend on the genetic background of strains derived from DC272 (adhC81) or MC1061. We found that DC272 carried a point mutation in the Cra-binding site of the adhE promoter. The Cra protein encoded by the cra gene is known to act as a repressor of adhE. P1-phage-mediated transduction and lacZ fusion analysis with the mutant adhE promoter confirmed that this mutation is responsible for overproduction. On the other hand, Southern hybridization revealed that MC1061 had a 0.85-kb deletion of the cra gene. Overproduction of AdhE in the MC1061 background was reversed to the wild-type levels by introduction of a plasmid carrying the cra(+) gene. These results indicated that expression of the adhE gene was regulated transcriptionally by Cra and posttranscriptionally by RNase G. PMID- 12083773 TI - Effect of infiltrated polyamines on polygalacturonase activity and chilling injury responses in zucchini squash (Cucurbita pepo L.). AB - The effect of exogenous polyamines on electrolyte leakage, chilling index, polygalacturonase activity (PG), ethylene production, and firmness in zucchini squash fruits stored for 12 days at 2 degrees C or 10 degrees C, 85-90% RH was evaluated. Fruits were infiltrated with putrescine (PUT) spermidine (SPD) and spermine (SPM) at 0.1, 0.25, 0.5, 2.0, and 4.0 mM. All polyamines exerted a protective effect on cell and organelle membranes. The most effective was SPD, which reduced electrolyte leakage between 62% and 82%, compared to control fruits stored at 2 degrees C. At 10 degrees C they did not exhibit chilling injury (CI) symptoms, while at 2 degrees C SPM (0.5 mM) and SPD (0.5 mM) diminished them 92% and 100%, respectively; which extended storage life for 8-10 days at 2 degrees C. High concentrations of polyamines (>2.0 mM) caused the appearance of CI symptoms. PG activity diminished proportionally to the concentration of polyamine except for the concentration at 4.0 mM. No significant changes were observed in ethylene production. PMID- 12083774 TI - Inhibition of glycogen synthase kinase-3beta by bivalent zinc ions: insight into the insulin-mimetic action of zinc. AB - Zinc is an important trace element found in most body tissues as bivalent cations and has essential roles in human health. The insulin-like effect of zinc cations raises the possibility that they inhibit glycogen synthase kinase-3beta (GSK 3beta), a serine/threonine protein kinase linked with insulin resistance and type 2 diabetes. Here we show that physiological concentrations of zinc ions directly inhibit GSK-3beta in vitro in an uncompetitive manner. Treatment of HEK-293 cells with zinc enhanced glycogen synthase activity and increased the intracellular levels of beta-catenin, providing evidence for inhibition of endogenous GSK-3beta by zinc. Moreover, zinc ions enhanced glucose uptake 3-fold in isolated mouse adipocytes, an increase similar to activation with saturated concentrations of insulin. We propose that the in vivo insulin-mimetic actions of zinc are mediated via direct inhibition of endogenous GSK-3beta. PMID- 12083775 TI - Behavior of fluorinated analogs of L-(3,4-dihydroxyphenyl)alanine and L-threo (3,4-dihydroxyphenyl)serine as substrates for Dopa decarboxylase. AB - We have determined the kinetic parameters for Dopa decarboxylase (DDC) of three ring-fluorinated analogs of 3,4-dihydroxyphenylalanine (Dopa). The rank order of catalytic efficiency of decarboxylation (k(cat)/K(m)) is Dopa>6-F-Dopa>2-F-Dopa>5 F-Dopa. This rank is consistent with previous in vivo and in vitro studies which indicate that, of the fluorinated analogs, 6-F-Dopa has pharmacokinetics that are most suited for positron emission tomographic (PET) evaluation of dopamine function. The effectiveness of PET as a diagnostic tool, the convenient half-life of (18)F (110 min) and the favorable pharmacokinetics of 6-[(18)F]FDOPA have combined to make this an extremely valuable reagent to study dopaminergic activity. The reactions of the related fluorinated DOPS analogs show that, while 6-F-threo-3,4-(dihydroxyphenyl)serine (DOPS) is decarboxylated at approximately the same rate as the non-fluorinated substrate, 2-F-threo-DOPS is not converted into the corresponding amine. In both cases a Pictet-Spengler condensation with the pyridoxal 5(')-phosphate (PLP) cofactor occurs to produce tetrahydroisoquinolines. Condensation of fluorinated catecholamines and catechol amino acids with endogenous aldehydes will be investigated as an approach to study possible mechanisms of L-Dopa-linked neurotoxicity. PMID- 12083776 TI - An extra human chromosome 21 reduces mlc-2a expression in chimeric mice and Down syndrome. AB - An extra copy of human chromosome 21 (Chr 21) causes Down syndrome (DS), which is characterized by mental retardation and congenital heart disease (CHD). Chimeric mice containing Chr 21 also exhibit phenotypic traits of DS including CHD. In this study, to identify genes contributing to DS phenotypes, we compared the overall protein expression patterns in hearts of Chr 21 chimeras and wild type mice by two-dimensional electrophoresis. The endogenous mouse atrial specific isoform of myosin light chain-2 (mlc-2a) protein was remarkably downregulated in the hearts of chimeric mice. We also confirmed that the human MLC-2A protein level was significantly lower in a human DS neonate heart, as compared to that of a normal control. Since mouse mlc-2a is involved in heart morphogenesis, our data suggest that the downregulation of this gene plays a crucial role in the CHD observed in DS. The dosage imbalance of Chr 21 has a trans-acting effect which lowers the expression of other genes encoded elsewhere in the genome. PMID- 12083777 TI - Identification of autoantibodies associated with systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the presence of antinuclear antibodies. We performed serological analysis of cDNA expression library (SEREX) to identify autoantibodies associated with SLE. The screening of three different cDNA expression libraries with pooled sera of patients with SLE yielded 11 independent clones that reacted with pooled sera of patients with SLE. In this screening, autoantibodies to poly(ADP-ribose) polymerase (PARP), U1snRNP, and galectin-3 were prevalent in the sera of patients with SLE (26/68, 25/68, 12/63, respectively). The frequency of autoantibody to PARP was significantly higher in SLE than that of healthy donors (0/76) (38.2% vs 0%, p<0.00001). The autoantibody to PARP was infrequently detected in the serum of patients with RA (1/50). However, autoantibody to PARP was not found in the sera of patients with other rheumatic diseases including Sjogren's syndrome (0/19), systemic sclerosis (0/18), and polymyositis/myositis (0/37). The frequency of autoantibody to human galectin-3 (12/63) was significantly higher in SLE than that of healthy donors (0/56) (19% vs 0%, p=0.0006). Autoantibody to galectin-3 was not found in the sera of patients with rheumatoid arthritis (0/50), Sjogren's syndrome (0/18), and systemic sclerosis (0/19). Interestingly, autoantibody to galectin-3 was also prevalent in the sera of patients with polymyositis/dermatomyositis (16/37, 43.2%). Further functional characterization of these autoantibodies would be necessary to determine their value as diagnostic markers or to define clinical subsets of patients with SLE. Statistical analysis revealed that the presence of autoantibody to PARP was inversely related with pleurisy, and the presence of autoantibody to galectin-3 related with renal disease. PMID- 12083778 TI - Purification and preliminary X-ray studies on hen serotransferrin in apo- and holo-forms. AB - Serum transferrins are monomeric glycoproteins with a molecular mass of around 80 kDa, that transport iron to cells via receptor-mediated endocytosis. Although both serum transferrins (STfs) and ovotransferrins (OTfs) are derived from the same gene in aves, the ovotransferrins do not transport iron in vivo. Crystal structures of OTf have been solved, in contrast no three-dimensional structure of avian STf have been determined as yet. Here we report the purification, crystallization, and preliminary crystallographic studies of the hen STf both in apo- (iron free) and holo- (iron loaded) forms. The hen STf has been purified to homogeneity by hydrophobic interaction chromatography. Both the apo- and holo forms were crystallized by hanging drop vapor diffusion method at 277 K. The apo crystals diffract to a resolution of 3.0 A and belong to the space group P4(3)2(1)2 with unit cell parameters a=b=90.5 and c=177.9 A. The holo-crystals diffract to a resolution of 2.8 A and belong to space group P2(1) with a=72.8, b=59.6, c=88.2 A, and beta=95.7 degrees. PMID- 12083779 TI - Cloning a cDNA encoding an alternatively spliced protein of BRCA2-associated factor 35. AB - Inheritance of mutations in the breast cancer susceptibility gene, BRCA2, predisposes humans to breast and ovarian cancers. Inherited mutations in the BRCA2 gene are also known to cause susceptibility to prostate cancer. BRCA2 protein exists in a large multi-protein complex from which a novel structural DNA binding protein BRCA2-associated factor 35 (BRAF35) has been isolated. We have cloned a novel cDNA encoding an alternatively spliced protein of BRAF35, designated as BRAF25. BRAF25 transcript is present in various human cells. We have precisely mapped the BRAF25 cDNA sequence to the genomic chromosome 19 sequence. Analysis of the predicted sequence of BRAF25 identified a protein of 215 amino acids. BRAF25 contains a truncated high mobility group domain, a kinesin-like coiled-coil domain and multiple Src homology 2 (SH2) motifs. Western blot analysis using antibodies specific for BRAF25 revealed the presence of BRAF25 in human prostate cancer cells. PMID- 12083780 TI - Immunohistological detection of BRAF25 in human prostate tumor and cancer specimens. AB - BRAF25 is an alternatively spliced protein of BRAF35 (see associated paper). We have mapped the BRAF25 gene to chromosome sub-band 19p13.3, a region where loss of chromosomal heterozygosity has been reported in about 50% of ovarian cancers. Because of the high incidence of genetic links of prostate cancer to breast and ovarian cancers, we investigated the BRAF25 expression in the prostate specimens. Immunohistochemical analysis using antibodies specific for BRAF25 revealed a strong immunostaining in sections of the benign prostatic hyperplasia (BPH). The staining was concentrated on the nuclei of cells facing the lumen of prostatic glands, even though the sporadic nuclei of cells in stromas were also stained. However, the expression of BRAF25 was dramatically reduced in intermediate prostate cancer and absent in advanced prostate cancer. Preincubation of the antibody with the immunizing peptide abolished immunostaining in BPH specimens. Therefore, the expression of BRAF25 was gradually lost in prostate cancer. PMID- 12083781 TI - Molecular model of shikimate kinase from Mycobacterium tuberculosis. AB - Tuberculosis (TB) resurged in the late 1980s and now kills approximately 3 million people a year. The reemergence of tuberculosis as a public health threat has created a need to develop new anti-mycobacterial agents. The shikimate pathway is an attractive target for herbicides and anti-microbial agents development because it is essential in algae, higher plants, bacteria, and fungi, but absent from mammals. Homologs to enzymes in the shikimate pathway have been identified in the genome sequence of Mycobacterium tuberculosis. Among them, the shikimate kinase I encoding gene (aroK) was proposed to be present by sequence homology. Accordingly, to pave the way for structural and functional efforts towards anti-mycobacterial agents development, here we describe the molecular modeling of M. tuberculosis shikimate kinase that should provide a structural framework on which the design of specific inhibitors may be based. PMID- 12083782 TI - Ile (476), a constituent of di-leucine-based motif of a major lysosomal membrane protein, LGP85/LIMP II, is important for its proper distribution in late endosomes and lysosomes. AB - Lysosomal membrane glycoprotein termed LGP85 or LIMP II extends a COOH-terminal cytoplasmic tail of R459GQGSMDEGTADERAPLIRT478, in which an L475 I476 sequence lies as a di-leucine-based motif for lysosomal targeting. In the present study, we explored the role of the I476 residue in the localization of LGP85 to the endocytic organelles using two substitution mutants called I476A and I476L in which alanine and leucine are replaced at I476, respectively, and I476R477T478 deleted LGP85 called Delta 476-478. Immunofluorescence analyses showed that I476A and I476L are largely colocalized in intracellular organelles with an endogenous late endosomal and lysosomal marker, LAMP-1, but there were some granules in which staining for the LGP85 mutants was prominent, while Delta 476-478 is detected in LAMP-1-positive and LAMP-1-negative intracellular organelles, and on the cell surface. The subcellular fractionation studies revealed that I476A, I476L, and Delta 476-478 are different from wild-type LGP85 in the distribution of early endosomes, late endosomes, and lysosomes. I476A and I476L are present more in late endosomes than in the densest lysosomes, whereas wild-type LGP85 is mainly lysosomal. Substitution of I476 for A and L differentially modified the ratios of late endosomal to lysosomal LGP85. A major portion of Delta 476-478 resided in the light buoyant density fraction containing plasma membrane and early endosomes. Taken together, these results indicate that the existence of the 476th amino acid residue is essential for localization of LGP85 to late endocytic compartments. The fact that isoleucine but not leucine is in the 476th position is especially of importance in the proper distribution of LGP85 in late endosomes and lysosomes. PMID- 12083783 TI - TIRAP mediates endotoxin-induced NF-kappaB activation and apoptosis in endothelial cells. AB - Bacterial lipopolysaccharide (LPS) initiates multiple signaling events in vascular endothelial cells that can result in activation and/or cell death. LPS induced activation of endothelial cells elicits a wide array of vascular endothelial responses, many of which are dependent on NF-kappaB activation. Several of the signaling molecules that mediate LPS-induced NF-kappaB activation, including Tlr-4, MyD88, and IRAK-1, have been similarly reported to mediate LPS pro-apoptotic signaling. Recently, a new signaling molecule, TIRAP, has been identified that mediates LPS-induced NF-kappaB signaling in monocytes and macrophages. Using a TIRAP dominant negative construct, we have identified a role for TIRAP in mediating LPS-induced NF-kappaB activation and apoptosis in human endothelial cells. These data identify TIRAP as a dual functioning signaling molecule and suggest the presence of a MyD88-independent LPS signaling pathway in human endothelial cells. PMID- 12083784 TI - Secretory production of recombinant human C-reactive protein in Escherichia coli, capable of binding with phosphorylcholine, and its characterization. AB - Recombinant human CRP (rhCRP) was secreted into culture supernatant of Escherichia coli by co-expressing kil gene that has a function to secrete colicin E1 outside the cell. Highly purified 5 g rhCRP was produced from 180 L culture supernatant by affinity chromatography. The purified rhCRP was indistinguishable from the native one with respect to Ca(2+)-dependent binding ability to phosphorylcholine, electrophoretic behavior, N-terminal amino acid analysis, and immunochemical properties. The molecular weight of rhCRP monomer was determined to be 23059.7 Da by TOF/MS analysis. These results indicate that rhCRP has the same protein structure as native one and that rhCRP has the potential as a reference material and/or calibrator of high-sensitivity CRP assay to predict the risk of cardiovascular disease. PMID- 12083785 TI - Structural characterization of Escherichia coli sialic acid synthase. AB - Sialic acid synthase encoded by the neuB gene of Escherichia coli catalyzes the condensation of N-acetylmannosamine and phosphoenolpyruvate to form N acetylneuraminic acid. This report demonstrates the first structural information on sialic acid synthase by CD, MALDI-TOF, and chemical cross-linking studies. Also, a specific cleavage by endogenous protease(s) has been identified at Lys(280) of the enzyme (40 kDa) by LC-MS and N-terminal sequencing analyses. The cleavage results in the formation of two inactive fragments of 33 and 7 kDa. The structural analysis indicates that the fragmentation is associated with a significant change of the enzyme from a tetrameric to trimeric form, and alterations in both secondary and native quaternary structures. PMID- 12083786 TI - The mif gene is transcriptionally regulated by glucose in insulin-secreting cells. AB - Macrophage migration inhibitory factor (MIF) is an important regulator of glucose homeostasis. In pancreatic beta-cells, MIF expression is regulated by glucose and its secretion potentiates the glucose-induced insulin secretion. The molecular mechanisms by which glucose mediates its effect on MIF expression are not elucidated. Herein, we report that incubating the differentiated insulin secreting cell line INS-1 in high glucose concentration increases MIF transcriptional activity as well as the reporter gene activity driven by the 1033 to +63 bp fragment of the MIF promoter. A minimal region located between 187 and -98 bp of this promoter sequence contributes both to basal activity and glucose-responsiveness of the gene. Within this promoter region, two cis-binding sequences were identified by mobility shift assays and footprinting experiments. Both cis-elements interact with nuclear proteins expressed specifically in insulin-secreting cells. In conclusion, we identified a minimal region of the MIF promoter which contributes to the glucose stimulation of the mif gene in insulin secreting cells. PMID- 12083787 TI - Particulate methane monooxygenase from Methylosinus trichosporium is a copper containing enzyme. AB - Particulate methane monooxygenase (pMMO) has been exfoliated and isolated from membranes of the Methylosinus trichosporium IMV 3011. It appears that the stability of pMMO in the exfoliation process is increased with increasing copper concentration in the growth medium, but extensive intracytoplasmic membrane formed under higher copper concentration may inhibit the exfoliation of active pMMO from membrane. The highest total activity of purified pMMO is obtained with an initial concentration of 6 microM Cu in the growth medium. The purified MMO contains only copper and does not utilize NADH as electron donor. Treatment of purified pMMO with EDTA resulted in little change in copper level, suggesting that the copper in the pMMO is tightly bound with pMMO. PMID- 12083788 TI - Effects of histone acetylation on transcriptional regulation of manganese superoxide dismutase gene. AB - To better understand the link between chromatin modification and manganese superoxide dismutase (Mn-SOD) gene expression, we have investigated the level of histone acetylation at Mn-SOD proximal promoter. TSA induced the expression of Mn SOD mRNA and its transcriptional activity in C2C12 cells. Sp1 binding sites in the proximal promoter region of Mn-SOD were transcriptionally responsive to TSA by transfection studies. We have detected a localized acetylation of histones H3 and H4, in vivo occupation by Sp1, early growth responsive-1 (Egr-1), and histone deacetylase-1 (HDAC1) in the proximal promoter region of Mn-SOD gene using chromatin immunoprecipitation assays. Our findings indicate that Mn-SOD gene expression is repressed by Sp1-HDAC1 complex. This repression is released by a localized histone acetylation and at least in parts a displacement by Egr-1 in response to TSA. PMID- 12083789 TI - Neonatal exposure to endocrine disruptors suppresses juvenile testis weight and steroidogenesis but spermatogenesis is considerably restored during puberty. AB - Neonatal exposure to endocrine disruptors induces developmental abnormalities in the male reproductive system. As to investigate whether neonatal exposure affects spermatogenesis in juvenile and pubertal testes, Sprague-Dawley rat pups were given various endocrine disruptors by a single injection on the day of birth at concentrations ranging between 4 microM and 40 mM and sacrificed on day 21 (juvenile) or 50 (puberty). The testes were weighed and examined histologically at each stage. Further, the metabolites of steroidogenesis were analyzed using normal-phase high performance liquid chromatography. Neonatal exposure significantly reduced testis weights and steroid biosynthesis of juveniles, but they were highly restored at puberty. PMID- 12083790 TI - Importance of phosphate contacts for sequence recognition by EcoRI restriction enzyme. AB - We have studied the importance of charge and hydrogen-bonding potential of the phosphodiester backbone for binding and cleavage by EcoRI restriction endonuclease. We used 12-mer oligodeoxynucleotide substrates with single substitutions of phosphates by chiral methylphosphonates at each position of the recognition sequence -pGpApApTpTpCp-. Binding was moderately reduced between 4- and 400-fold more or less equally for the R(P) and S(P)-analogues mainly caused by missing charge interaction. The range of cleavage effects was much wider. Four substrates were not cleaved at all. At both flanking positions and in the purine half of the sequence up to the central position, cleavage was more impaired than binding and differences between R(P) and S(P) diastereomeres were more pronounced. These effects are easily interpreted by direct phosphate contacts seen in the crystal structure. For the effects of substitutions in the pyrimidine half of the recognition sequence, more indirect effects have to be discussed. PMID- 12083791 TI - Emergent literacy skills and training time uniquely predict variability in responses to phonemic awareness training in disadvantaged kindergartners. AB - The factors that predicted variability in responses to phonemic awareness training were investigated in kindergartners who live in poverty. Treatment children (n=42) received both analytic and synthetic phonemic awareness computer assisted instruction, while controls (n=34) received no special training. Mean age of participants was approximately 5 years 7 months. Pretests included initial phonemic awareness, letter knowledge, word-level reading, invented spelling, vocabulary knowledge, and print concepts. Spelling skills emerged as the best consistent predictor of variability in phonemic awareness in response to instruction. We propose that relations between phonemic awareness and spelling skills are bidirectional: Spelling influenced growth in phonemic awareness and phonemic awareness contributed to growth in spelling skills. The amount of exposure that children had to the treatment intervention contributed uniquely to individual differences in posttest levels of phonemic awareness and spelling. PMID- 12083792 TI - A longitudinal study of the development of shifts of gaze to a peripheral stimulus in preterm infants with transient periventricular echogenicity. AB - Shifts of gaze to peripheral targets were studied longitudinally, between 6 and 26 weeks corrected age, in full-term and very preterm infants with transient periventricular echogenicity (PVE). Before 10 weeks, simple shifts of gaze were faster and more frequent in preterms with PVE<14 days than in full-terms, suggesting these preterms profited from additional early visual experience. After 16 weeks, there were subtle differences between full- and preterm infants in the development of shifts of gaze requiring disengagement. The differences suggest that, after disengagement had become established, its fine-tuning occurred more slowly in the preterms. Slower fine-tuning of disengagement was not associated with duration of PVE, since it was more marked in preterms with PVE<14 days than in preterms with PVE> or =14 days. The differences in performance between full- and preterm infants were small. However, even small differences may affect the efficiency of visually guided behaviors. PMID- 12083793 TI - The development of exogenous orienting: mechanisms of control. AB - The present study examined exogenous orienting in children ages 6 years (n=14), 10 years (n=13), 14 years (n=14), and adults (n=13). The main purpose was to identify which of the attentional operations underlying exogenous orienting (disengaging, shifting, and/or engaging) improves with age. A simple detection task was used in which single targets were correctly (80%) or incorrectly (20%) cued. The main finding was that disengaging attention alone distinguished the performance of younger children from that of older children and adults, regardless of whether attention alone (covert attention) or attention and associated sensory and motor systems (overt attention) were involved. In addition, our data, in contrast to those reported previously, raise the possibility that even the youngest children tested were able to use the predictability of the cue to enhance their performance. These and related neuroanatomical findings are taken as evidence for the development with age of more flexible control in orienting visual attention. PMID- 12083794 TI - On the nature of the relationship between processing activity and item retention in children. AB - The concept of working memory emphasizes the interrelationship between the transient retention of information and concurrent processing activity. Three experiments address this relationship in children between 8 and 17 years of age by examining forgetting when a processing task is interpolated between presentation and recall of the memory items. Unlike previous studies, delivery of interpolated stimuli was under computer control and responses to these stimuli were timed. There were consistent effects of the duration of the interpolated task, but no effects of either its difficulty or similarity to memory material and no qualitative developmental differences in task performance. The absence of an effect of difficulty provides no support for models of working memory in which limited capacity is shared between the dual functions of processing and storage, but is compatible with an alternative "task switching" account. However, task switching did not explain developmental differences in recall. Other aspects of the results suggest that there can be interactions between processing and storage but it is argued that these cannot be straightforwardly explained in terms of either task switching or resource sharing. PMID- 12083795 TI - Identification of myo-inositol 1,2-cyclic monophosphate by electrospray tandem mass spectrometry, a major constituent of EGF-stimulated phosphoinositide turnover in MDA 468 cells. AB - Epidermal growth factor (EGF) caused an increase in phosphoinositide (PI) turnover in MDA 468 cells. This EGF-stimulated effect was inhibited by the protein tyrosine kinase inhibitor lavendustin A (LA). MDA 468 cells generated an atypical PI turnover profile. Examination and quantitation of the PI metabolite profile showed that even control cells produced a metabolite which was acid labile and which formed about 60% of the total PI metabolites. By using the technique of electrospray ionization tandem mass spectrometry, we were able to confirm the identity of this acid-labile metabolite through the specific fragmentation as compared with the standard. The precursor molecule fragmented into two distinct productions with molar masses identical to that of the standard myo-inositol 1,2-cyclic monophosphate (cInsP). Changes in the PI turnover profile could be accounted for by the alterations in myo-inositol 1,2-cyclic monophosphate generated in these cells. We thus conclude that, by some as-yet unidentified mechanism, cyclic inositol monophosphate forms a major constituent of EGF-stimulated PI turnover in MDA 468 cells. PMID- 12083796 TI - Mitogen-induced, FAK-dependent tyrosine phosphorylation of the SSeCKS scaffolding protein. AB - The ability of mitogens to rapidly induce tyrosine phosphorylation of cellular proteins has been taken as evidence of participation in subsequent signaling pathways. SSeCKS, a major protein kinase C (PKC) substrate with protein scaffolding and tumor suppressive properties, becomes tyrosine phosphorylated in NIH3T3 and rodent embryo fibroblasts after short-term treatment with epidermal growth factor (EGF), platelet-derived growth factor (PDGF), or fetal calf serum in the presence of pervanadate, but not by treatment with insulin or insulin-like growth factor-1. The relative phosphotyrosine level on SSeCKS was higher in actively dividing cells than in confluent cultures. Tyrosine phosphorylation of SSeCKS was apparent in cells deficient in Src, Fyn, Yes, or Abl tyrosine kinases or in NIH3T3 cells expressing a temperature-sensitive v-Src allele, but not in FAK-deficient embryo fibroblasts. Purified FAK or Src enzyme failed to directly phosphorylate SSeCKS in vitro. EGF failed to induce SSeCKS tyrosine phosphorylation in FAK-/- fibroblasts, indicating that the EGF receptor is probably not the direct kinase of SSeCKS. Phosphorylation under these conditions was rescued by the transient reexpression of wt-FAK but not FAK mutated at Y397, a major autophosphorylation and SH2-based docking site. Adhesion of FAK+/+ cells to fibronectin failed to significantly induce SSeCKS tyrosine phosphorylation although FAK was activated, suggesting that SSeCKS phosphorylation is mediated through a growth factor receptor-FAK rather than an integrin-FAK pathway. Moreover, PDGF could induce SSeCKS tyrosine phosphorylation in the absence of FAK activation, suggesting a role for FAK SH2-based docking rather than kinase activity. Immunofluorescence analysis showed that in FAK-/- cells, SSeCKS costains along F-actin stress fibers, in contrast to FAK+/+ cells, where most SSeCKS stains at the cell edge and along a cortical cytoskeletal matrix. This correlated with increased coprecipitation of SSeCKS with biotin-phalloidin-bound F-actin from FAK-/- compared to FAK+/+ cell lysates. Similarly, bacterially expressed, unphosphorylated SSeCKS cosedimented with F-actin in ultracentrifugation assays. These data suggest that mitogen-induced, FAK dependent tyrosine phosphorylation of SSeCKS modulates its binding to the actin based cytoskeleton, suggesting a role for SSeCKS in mitogen-induced cytoskeletal reorganization. PMID- 12083797 TI - The topoisomerase I-binding RING protein, topors, is associated with promyelocytic leukemia nuclear bodies. AB - We previously identified topors as a topoisomerase I-binding protein that localizes in punctate nuclear regions when expressed as a GFP fusion protein. We now demonstrate that both the GFP-topors fusion protein and endogenous topors are associated with promyelocytic leukemia (PML) nuclear bodies in exponentially growing HeLa cells. Studies using isogenic PML+/+ and PML-/- murine embryonic fibroblasts indicate that the punctate nuclear localization of topors is dependent on PML. A basic C-terminal region but not the N-terminal RING domain of topors is required for the punctate nuclear localization of this protein. Additional studies indicate that topors, but not PML, rapidly relocalizes from nuclear bodies to the nucleoplasm in cells exposed to the transcription inhibitor dichloro-1-beta-d-ribofuranolsylbenzimidazole or to the topoisomerase I-targeting drug camptothecin. These results identify topors as a new member of the group of proteins that associate dynamically with PML nuclear bodies and suggest that topors may be involved in the cellular response to camptothecin. PMID- 12083798 TI - Differences in cortical actin structure and dynamics document that different types of blebs are formed by distinct mechanisms. AB - Bleb formation has been studied by specifically targeting major factors controlling this process, such as microtubule disassembly, local actin depolymerization, and increased pressure. At least two different types of blebs (types 1 and 2) formed by different mechanisms and possibly a third type (type 3) can be documented at the front of living polarized cells expressing green fluorescent protein-actin and/or in fixed and stained cells. Type 1 blebs (membrane/cortex dissociation blebs) formed by dissociation of the plasma membrane from cortical actin develop cytoplasmic actin layers associated with restriction rings. They can be induced by the microtubule-disassembling agent colchicine. Type 2 blebs (cortical actin disassembly blebs) form after disassembly of the cortical actin layer in the presence of latrunculin A. Restriction rings without a cytoplasmic actin layer occur in a transition zone between the intact cortical actin layer of the cell body and the compromised actin layer of the bleb. Evidence for a third type of bleb (type 3), showing an intact cortical actin layer but no cytoplasmic actin layer and no recognizable relationship between the actin cytoskeleton and the restriction ring, has been obtained by passive cell deformation in micropipettes, which increases pressure. Repolymerization of the cortical actin layer does not necessarily result in bleb retraction. Once formed, restriction rings do not narrow, suggesting that they result from isometric contraction. A simplified classification scheme has been developed to relate the type of bleb to specific signals or cell functions. Its application shows that spontaneously blebbing cells form almost exclusively type 1 blebs. PMID- 12083799 TI - Induction of senescence-like state and suppression of telomerase activity through inhibition of HPV E6/E7 gene expression in cells immortalized by HPV16 DNA. AB - The E6 and E7 oncoproteins of human papillomavirus (HPV) play a major role in the development of cervical carcinoma. In this study, a recombinant adenovirus that expresses the bovine papillomavirus (BPV) E2, which has been shown to inhibit HPV early gene expression, was delivered to two HPV-immortalized cell lines as well as CaSki, a cervical carcinoma cell line. We tested whether the carcinoma and the immortal cells were equally affected by the expression of BPV E2. In all cell lines, BPV E2-mediated inhibition of HPV E6/E7 expression caused a dramatic suppression of cell growth, being preceded by the activation of the p53-Rb growth inhibitory pathway, and a decrease in hTERT mRNA expression and telomerase activity. This suggests that the HPV E6 and E7 proteins are required not only for induction of the proliferative phenotype and telomerase activity, but also for their maintenance. In both the carcinoma and the immortal lines, the number of cells with enlarged cytoplasm and senescence-associated beta-galactosidase activity, which are markers for cellular senescence, was significantly increased. These results suggest that a senescence program exists in cells immortalized by HPV DNA as well as in cervical carcinoma cells. PMID- 12083800 TI - Translocation of the classic protein kinase C isoforms in porcine oocytes: implications of protein kinase C involvement in the regulation of nuclear activity and cortical granule exocytosis. AB - Protein kinase C (PKC) is a family of Ser/Thr protein kinases categorized into three subfamilies: classical, novel, and atypical. The subcellular localization of classical PKCalpha, -betaI, and -gamma in the process of porcine oocyte maturation, fertilization, and parthenogenetic activation and their involvement in cortical granule (CG) exocytosis were investigated. The results of Western blot showed that PKCalpha, -betaI, and -gamma were expressed in the oocytes at the germinal vesicle (GV) and metaphase II (MII) stages. Confocal microscopy revealed that the three PKC isoforms were concentrated in the GV but evenly distributed in the cytoplasm of MII eggs. PKCalpha and -gamma were translocated to the plasma membrane soon after sperm penetration. cPKCs migrated into the pronucleus in fertilized eggs. Following treatment with a PKC activator, phorbol 12-myristate 13-acetate (PMA), CGs were released and PKCalpha and -gamma were translocated to the membrane. The CG exocytosis and PKC redistribution induced by PMA could be blocked by the PKC inhibitor staurosporine. Parthenogenetic stimulation with ionophore A23187 or electrical pulse also induced cPKC translocation and CG exocytosis. Eggs injected with PKCalpha isoform-specific antibody failed to undergo CG exocytosis after PMA treatment or fertilization. The results suggest that cPKCs, especially the alpha-isotype, regulate nuclear function and CG exocytosis in porcine eggs. PMID- 12083801 TI - Superoxide anion-dependent Raf/MEK/ERK activation by peroxisome proliferator activated receptor gamma agonists 15-deoxy-delta(12,14)-prostaglandin J(2), ciglitazone, and GW1929. AB - In this study, we examined the signaling pathways for extracellular signal related protein kinase (ERK) activation by three structurally different peroxisome proliferator activated receptor-gamma (PPARgamma) agonists. In murine C2C12 myoblasts, treatment with 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d PGJ(2)), ciglitazone, and GW1929 leads to ERK1/2 phosphorylation in a time- and concentration-dependent manner. Consistent with ERK phosphorylation, mitogen activated protein/ERK kinase (MEK) phosphorylation as well as Raf-1 kinase activity are also accordingly stimulated, while the constitutive Ser259 phosphorylation of Raf-1 is decreased. The ERK phosphorylation induced by PPARgamma agonists is not blocked by the PKC inhibitors GF109203X and Ro31-8220, the PI3K inhibitor wortmannin, the Ras inhibitor FPTI, the negative mutant of Ras, or the PPARgamma antagonist bisphenol A diglycidil ether. Expression of PPARgamma2 without DNA binding domain or with a nonphosphorylatable mutant (S112A) fails to change ERK phosphorylation by 15d-PGJ(2). On the contrary, the ERK phosphorylation by PPARgamma agonists is inhibited by the MEK inhibitor PD98059, GSH, and permeable SOD mimetic MnTBAP. Chemiluminescence study reveals that these three PPARgamma agonists are able to induce superoxide anion production, with an efficacy similar to their action on ERK phosphorylation. Consistent with this notion, we also show that superoxide anion donor 2,3 dimethoxy-1,4-naphoquinone elicits ERK phosphorylation. In this study, we for the first time demonstrate a novel mechanism, independent of Ras activation but initiated by superoxide anion production, for PPARgamma agonists to trigger the Raf-MEK-ERK1/2 signaling pathway. PMID- 12083802 TI - Nucleolar localization of hTERT protein is associated with telomerase function. AB - Telomerase is a ribonucleoprotein (RNP) complex that prevents telomeric erosion in eukaryotic cells. Although there are also other associated proteins in this complex, the catalytic activity of this complex is composed of two components. One is a reverse transcriptase subunit, TERT (telomerase reverse transcriptase); another is an RNA template subunit, TR (telomerase RNA). However, where these two parts are assembled in mammalian cells is unclear. In the present study, we investigated the intracellular distribution of human TERT (hTERT) protein and observed that hTERT protein in individual cells could concentrate in or be excluded from the nucleolus. Further we have identified a nucleolar targeting signal in the hTERT protein. Point mutations that disrupted this signal region interrupted telomerase RNP complex formation, decreased telomerase activity, and caused telomere shortening in cells transfected with mutated hTERT. Our results indicate that the amino acid sequence of the extreme N-terminus (1-15) of hTERT, which targets nucleolar localization of the protein, is required for full telomerase function. PMID- 12083803 TI - Procathepsin D interacts with prosaposin in cancer cells but its internalization is not mediated by LDL receptor-related protein. AB - The cell surface binding, endocytosis, and lysosomal routing of procathepsin D (procath-D) in cancer cells are mostly independent of the mannose-6-phosphate (M6P) receptors. In an attempt to define the receptor involved, we intracellularly cross-linked procath-D with a 68-kDa protein that we identified with specific antibodies as prosaposin in human breast and ovarian cancer cell lines. In cancer cells, this protein-protein interaction was resistant to ammonium chloride or M6P treatment, indicating that it was independent of the M6P receptors. A similar interaction also occurred in the breast cancer cell culture medium between the secreted prosaposin and procath-D. Since these two precursors can be endocytosed, we then determined whether they were interacting with the same cell surface receptor. In fibroblasts, we confirmed that the endocytosis of these two proteins was different since it was generally mediated by the M6P receptors for procath-D and mostly by LRP (LDL receptor-related protein) for prosaposin. In breast cancer cells, prosaposin endocytosis was not detected, in contrast to procath-D endocytosis, suggesting that the majority of procath-D is not internalized as a complex with prosaposin. Moreover, RAP (receptor-associated protein), a ligand inhibiting LRP-mediated endocytosis, prevented internalization of prosaposin in 49-F rat fibroblasts, but did not affect procath-D M6P independent internalization in MDA-MB231 cells. We conclude that in breast cancer cells, even though procath-D interacts intracellularly and extracellarly with prosaposin, it is endocytosed independent of prosaposin by a receptor different from the M6P receptors and the LRP. PMID- 12083804 TI - Necdin is required for terminal differentiation and survival of primary dorsal root ganglion neurons. AB - Necdin is expressed predominantly in postmitotic neurons and serves as a growth suppressor that is functionally similar to the retinoblastoma tumor suppressor protein. Using primary cultures of dorsal root ganglion (DRG) of mouse embryos, we investigated the involvement of necdin in the terminal differentiation of neurons. DRG cells were prepared from mouse embryos at 12.5 days of gestation and cultured in the presence of nerve growth factor (NGF). Immunocytochemistry revealed that necdin accumulated in the nucleus of differentiated neurons that showed neurite extension and expressed the neuronal markers microtubule associated protein 2 and synaptophysin. Suppression of necdin expression in DRG cultures treated with antisense oligonucleotides led to a marked reduction in the number of terminally differentiated neurons. The antisense oligonucleotide treated cells did not attempt to reenter the cell cycle, but underwent death with characteristics of apoptosis such as caspase-3 activation, nuclear condensation, and chromosomal DNA fragmentation. Furthermore, a caspase-3 inhibitor rescued antisense oligonucleotide-treated cells from apoptosis and significantly increased the population of terminally differentiated neurons. These results suggest that necdin mediates the terminal differentiation and survival of NGF dependent DRG neurons and that necdin-deficient nascent neurons are destined to caspase-3-dependent apoptosis. PMID- 12083805 TI - The role of SF1 in adrenal and reproductive function: insight from naturally occurring mutations in humans. AB - Steroidogenic factor 1 is a monomeric orphan nuclear receptor and one of several hundreds of transcription factors encoded in the human genome. It regulates the transcription of many genes involved in gonadal development, sexual differentiation, steroidogenesis and reproduction. Recently, mutations in the gene encoding SF1 have been identified in several patients with primary adrenal failure and 46,XY sex-reversal. Interpreting the consequences of these mutations provides further understanding of transcription factor haploinsufficiency in human genetic disease as well as the exquisite sensitivity of humans to gene dosage effects during adrenal and gonadal development. PMID- 12083806 TI - Upregulation of smooth muscle cell collagen production by homocysteine-insight into the pathogenesis of homocystinuria. AB - Patients with untreated homocystinuria have widespread premature atherosclerosis with intimal thickening and collagen-rich, fibrous plaques. We previously demonstrated that homocysteine (Hcy) upregulates collagen synthesis and accumulation by arterial smooth muscle cells (SMCs) [A. Majors, L.A. Ehrhart, E.H. Pezacka, Arterioscler. Thromb. Vasc. Biol. 17 (1997) 2074-2081] but the underlying mechanisms are not known. Since many of the effects of Hcy on intact vessels and vascular cells are thought to involve reactive oxygen species generated from Hcy oxidation, we investigated the role of reactive oxygen species in the upregulation of collagen production by Hcy. Treatment of SMCs with 300 microM l-Hcy increased collagen accumulation 2-3-fold. When added to culture medium containing serum, the exogenous Hcy was rapidly oxidized with a half-life of approximately 1 h but only very low amounts of H(2)O(2) (up to 2 microM) were detected. Three lines of evidence demonstrate that the increased accumulation of collagen was not mediated by reactive oxygen species generated from Hcy oxidation: (1) catalase in the medium did not block the accumulation of collagen in Hcy-treated cultures; (2) the addition of xanthine/xanthine oxidase, a system that generates superoxide and H(2)O(2), did not increase collagen accumulation; and (3) the direct addition of H(2)O(2) did not substantially enhance collagen accumulation. In contrast, heparin, a potent modulator of SMC function, significantly blocked the accumulation of collagen in Hcy-treated cultures. Together, these results demonstrate that the increase in collagen accumulation in Hcy-treated cultures involves alternate mechanisms not involving H(2)O(2). PMID- 12083807 TI - Expression of the liver form of arginase in erythrocytes. AB - Arginase I (AI) has a critical function in mammalian liver as the final enzyme in the urea cycle responsible for the disposal of ammonia from protein catabolism. AI is also expressed in various extrahepatic tissues and may play a role in regulating arginine levels and in providing ornithine for biosynthetic reactions that generate various critical intermediary metabolites such as glutamate, glutamine, GABA, agmatine, polyamines, creatine, proline, and nitric oxide. AI is expressed in red blood cells (RBCs) only in humans and certain higher primates. Macaca fascicularis has been identified as an evolutionary transition species in which RBC-AI expression is co-dominantly regulated. The M. fascicularis AI gene was analyzed to understand AI expression in erythrocytes. Erythroid progenitor cells [nucleated red blood cells (nRBCs)] isolated from cord blood were utilized to demonstrate AI expression by immunocytochemical staining using anti-AI antibody. Introduction of EGFP reporter vectors into nRBC showed that the proximal 1.2 kbp upstream of the AI gene is sufficient for AI expression. Expression of a second arginase isoform, AII, in nRBCs was discovered by cDNA profiling. This contrasts with mature fetal or adult RBCs which contain only the AI protein. In addition, an alternatively spliced AI (AI(')) variant was observed from erythroid mRNA analysis with an alternative splice acceptor site located within intron 2, causing the insertion of eight additional amino acids yet retaining significant enzymatic activity. PMID- 12083808 TI - Identification and cloning of a new protein that binds the 3(') untranslated region of alpha-striated tropomyosin. AB - The 3' untranslated region of muscle tropomyosin (TM UTR) induces muscle differentiation when transcribed in primary fibroblasts. This sequence binds protein in extracts from cell types that differentiate upon TM UTR transcription. To identify the protein(s) bound by the TM UTR, an avian embryo fibroblast library was induced to express protein in solution and extracts from these pools were screened with electromobility shift assays using a TM UTR RNA probe. Positive pools were progressively fractionated until a pool containing a single positive clone was obtained. The TM UTR-binding protein (UBP) clone thus isolated contains 751 nt, 618 of which represent a single open reading frame. UBP is related to a human autoantigen, Sjogren's syndrome antigen B (SSB) beginning with the start of the UBP open reading frame. This homology is to the 5' end of SSB in a region containing an RNA-binding motif of 70 amino acids. The deduced amino acid sequence of UBP predicts phosphorylation sites for protein kinase C, casein kinase 2, and cAMP-dependent protein kinase and asparginine glycosylation sites. The observed size of UBP by UV cross-linking with a TM UTR probe is of the same size as the protein bound in fibroblast extract. UBP is expressed in primary fibroblasts, but not in fibroblast or myogenic cell lines, suggesting that its expression is restricted. The full-length UBP mRNA is approximately 3 kB, suggesting a long 5' untranslated region. Transient transfection of cultured cells with UBP directs production of a protein that binds the TM UTR, confirming that these sequences interact in vivo. These observations suggest that we have identified a novel protein that binds to the TM UTR in vitro and in vivo. Determining the function of this protein will facilitate determining the mechanism by which the TM UTR induces differentiation. PMID- 12083809 TI - Base composition at mtDNA boundaries suggests a DNA triple helix model for human mitochondrial DNA large-scale rearrangements. AB - Different mechanisms have been proposed to account for mitochondrial DNA (mtDNA) instability based on the presence of short homologous sequences (direct repeats, DR) at the potential boundaries of mtDNA rearrangements. Among them, slippage mispairing of the replication complex during the asymmetric replication cycle of the mammalian mitochondrial DNA has been proposed to account for the preferential localization of deletions. This mechanism involves a transfer of the replication complex from the first neo-synthesized heavy (H) strand of the DR1, to the DR2, thus bypassing the intervening sequence and producing a deleted molecule. Nevertheless, the nature of the bonds between the DNA strands remains unknown as the forward sequence of DR2, beyond the replication complex, stays double stranded. Here, we have analyzed the base composition of the DR at the boundaries of mtDNA deletions and duplications and found a skewed pyrimidine content of about 75% in the light-strand DNA template. This suggests the possible building of a DNA triple helix between the G-rich neo-synthesized DR1 and the base-paired homologous G.C-rich DR2. In vitro experiments with the purified human DNA polymerase gamma subunits enabled us to show that the third DNA strand may be used as a primer for DNA replication, using a template with the direct repeat forming a hairpin, with which the primer could initiate DNA replication. These data suggest a novel molecular basis for mitochondrial DNA rearrangements through the distributive nature of the DNA polymerase gamma, at the level of the direct repeats. A general model accounting for large-scale mitochondrial DNA deletion and duplication is proposed. These experiments extend to a DNA polymerase from an eucaryote source the use of a DNA triple helix strand as a primer, like other DNA polymerases from phage and bacterial origins. PMID- 12083810 TI - Pilot study of mass screening for Wilson's disease in Korea. AB - Wilson's disease (WD) is an autosomal recessive disorder of copper metabolism with copper accumulation in the liver as well as in the central nervous system. Treatment of WD includes oral chelating agents and diet and it is effective. However, once irreversible damage has occurred, the effect of treatment is diminished and the patient's quality of life is compromised. It is estimated that at least half of the patients with WD remain undiagnosed and die of untreated disease. Early detection of patients presymptomatically has been hampered by the lack of effective methods for mass screening. Recently, a sandwich ELISA method for ceruloplasmin measurement in blood spots was developed. We have used this method to analyze blood specimens collected on filter paper from 3667 children aged 3 months-15 years. The mean value of ceruloplasmin was 30.5+/-9.5 mg/dL. Among these children, we identified one WD case, a 32-month-old boy with markedly reduced ceruloplasmin concentration (2.3 mg/dL). Measurement of CP level in dried blood spot sample is proposed as a reliable method for population screening of WD. PMID- 12083811 TI - Genetic analysis in nine unrelated Italian patients affected by OTC deficiency: detection of novel mutations in the OTC gene. AB - Ornithine transcarbamylase deficiency (OTCD) is an X-linked urea cycle disorder due to a defect of the mithocondrial enzyme ornithine transcarbamylase (OTC). Genetic analysis in nine unrelated Italian patients affected by OTCD (one male patient and eight female manifesting carriers) led to the detection of three novel mutations and six previously reported mutations in the OTC gene. The analysis was performed by direct sequencing of OTC cDNA, OTC exons, and intron exon boundaries and enzymatic restriction analysis on the patients' genomic DNA and total RNA isolated from peripheral blood lymphocytes. In the male patient the new mutation S132P due to the nucleotide change c.394T>C was identified. In a manifesting carrier the nucleotide change c.292G>A that leads to the novel amino acid substitution E98K was identified; this mutation is close to the OTC protein's carbamyl phospate binding site. In another manifesting carrier the OTC cDNA analysis revealed the normally spliced transcript and an aberrant transcript with an insertion of two nucleotides (c.77-78insAG). In the patient's genomic DNA we identified a new transvertion IVS1-3C>G at the heterozygous state; this nucleotide change generates a new splice acceptor site in intron 1 that induces an RNA splicing defect. This insertion causes a frame shift in OTC cDNA ORF and leads to a premature stop codon. The previously described mutations N161S, R141Q, T178M, R92X, A208T, M268T were identified in the other six manifesting carriers. PMID- 12083812 TI - Identification of a novel cathepsin C mutation (p.W185X) in a Brazilian kindred with Papillon-Lefevre syndrome. AB - Papillon-Lefevre syndrome (PLS) is an autosomal recessive palmoplantar keratoderma caused by cathepsin C (CTSC) gene mutations. This study reports CTSC mutational and enzyme analyses in a consanguineous Brazilian family with PLS, representing the first enzymatic analysis of a Brazilian kinship with PLS. This family segregates a novel PLS-related mutation, p.W185X, that is associated with a complete loss of enzymatic activity. PMID- 12083813 TI - The role of the HNF4alpha enhancer in type 2 diabetes. AB - The genetic causes of type 2 diabetes are not well understood. The disease has been linked to chromosome 20q12-q13.1 a region which harbors the transcription factor HNF4alpha. Mutations in the coding region of HNF4alpha cause maturity onset diabetes of the young, an autosomal dominant form of diabetes, but do not account for the linkage to this region. An enhancer element has recently been characterized 6 kb 5' of the HNF4alpha P1 promoter containing binding sites for the transcription factors HNF1, HNF4, HNF3, and C/EBP, which are overlapped by glucocorticoid consensus sites. We hypothesized that variation in the enhancer element disrupts HNF4alpha expression in the liver and increases susceptibility to type 2 diabetes. We screened for variants of the enhancer element in 39 white UK young onset diabetic subjects, giving >95% power to identify variants with minor allele frequencies of >5%. No variants of the enhancer element were found in this population. We conclude that variation in the HNF4alpha enhancer element is not a common cause of susceptibility to type 2 diabetes. PMID- 12083814 TI - No evidence for involvement of the calpain-10 gene 'high-risk' haplotype combination for non-insulin-dependent diabetes mellitus in early onset obesity. AB - In light of evidence of linkage of obesity to chromosome 2q31-q37, we hypothesized that the calpain-10 gene 'high-risk' haplotype combination for non insulin-dependent diabetes mellitus (NIDDM) is involved in early onset obesity. We screened the NIDDM 'high-risk'-haplotype combination formed by the alleles 112 and 121 of the polymorphisms UCSNP-43, -19, and -63 in 166 families consisting of an extremely obese child or adolescent (mean BMI percentile: 99.3+/-1.38), one or more obese sibs (mean BMI percentile: 97.42+/-2.88), and both of their parents. Genotyping for three calpain-10 gene polymorphisms was performed by polymerase chain reaction (PCR) with (a) length polymorphism detection (UCSNP-19) or (b) allele-specific PCR (UCSNP-43 and -63). To allow for correct haplotype assignment all individuals were additionally genotyped for two microsatellite markers (D2S125 and D2S2338). We followed a hierarchical test procedure. As the first step, model-free linkage analysis was performed using maximum likelihood binomial statistics. The second stage consisted of a one-sided asymptotic pedigree disequilibrium test for the UCSNP-43 and on an exploratory level for the other SNP-markers and all haplotypes formed by the three SNPs. The final stage investigated the reported haplotype combination. We failed to detect an initial linkage of obesity to this region (LOD score <0.4). All subsequent exploratory analyses were negative. Our analysis of the relationship between the NIDDM 'high risk' haplotype combination and extreme early onset obesity revealed no evidence for linkage and association. PMID- 12083815 TI - Adrenal hypoplasia congenita with multiple pituitary hormone deficiency without documented mutation in DAX1 or SF1 gene. AB - A boy with adrenal hypoplasia congenita (AHC) and multiple pituitary hormone deficiency (MPHD), without mutations in the DAX1 or SF1 genes, is described. The association of AHC and MPHD has not been previously reported. PMID- 12083816 TI - Induction of alternatively spliced spermidine/spermine N1-acetyltransferase mRNA in the human kidney cells infected by venezuelan equine encephalitis and tick borne encephalitis viruses. AB - 293 and RH cells derived from human embryo kidney were infected by Venezuelan equine encephalitis and tick-borne encephalitis viruses and cDNA libraries representing cellular mRNAs induced or suppressed due to the infection were prepared using suppressive subtractive hybridization. Among the up-regulated clones the RT-PCR and Northern analyses revealed an unusual transcript of the spermidine/spermine N1-acetyltransferase (SSAT) gene that was shown to be an alternatively spliced form containing an additional 110-bp exon. The alternatively spliced transcript is polyadenylated and can be expected to yield only a truncated 71 amino acid polypeptide. This first evidence of the host gene alternatively spliced mRNA induction by RNA viruses raises the questions of its biological role, regulation mechanisms of alternative splicing, and significance for the virus life cycle. PMID- 12083818 TI - Hepatitis C virus IRES-dependent translation is insensitive to an eIF2alpha independent mechanism of inhibition by interferon in hepatocyte cell lines. AB - Interferon (IFN) in combination with ribavirin is the main treatment for hepatitis C virus (HCV) infection. The sensitivity or resistance of the virus to IFN has been linked to certain types of the interferon sensitivity determining region (ISDR) and PKR-eIF2alpha phosphorylation homology domain (PePHD) sequences in the NS5A and E2 regions of the viral genome, respectively. In search of the other potential mechanisms of HCV resistance to IFN, we tested the effect of IFN alpha on translational activity of the HCV IRES in various cell types. Using bicistronic dual luciferase reporter RNAs in direct RNA transfection studies, we found that the cap-dependent translation was dramatically inhibited by IFN (5- to 16-fold), whereas HCV IRES translation was inhibited only marginally in two hepatoma cell lines, Huh7 and HepG2 cells. No difference in IFN sensitivity was observed among IRESs of genotypes 1a, 1b, and 2a. Translation under the control of encephalomyocarditis virus (EMCV) IRES was inhibited by IFN to the same extent as cap-dependent translation. In cells of nonhepatic origin (HeLa and Raji), however, HCV IRES-, EMCV IRES-, and cap-dependent translation were dramatically inhibited to similar levels. The PKR expression level was enhanced by IFN in all cells, but eIF2alpha phosphorylation level was not changed, probably due to the absence of double-stranded RNA species. There was also no evidence of RNase L activation. Therefore, inhibition of translation by IFN under these conditions was probably mediated by novel IFN-induced inhibitory pathways, independent of eIF2alpha phosphorylation, while HCV IRES was not subject to this inhibition in hepatoma cell lines. Thus, HCV IRES-driven translation was resistant to IFN induced, eIF2alpha-independent inhibition in human hepatoma cells that are frequently used in studies on HCV replication. This may present a new potential mechanism of viral resistance to IFN treatment during the early steps of virus infection. PMID- 12083817 TI - Analysis of the monkeypox virus genome. AB - Monkeypox virus (MPV) belongs to the orthopoxvirus genus of the family Poxviridae, is endemic in parts of Africa, and causes a human disease that resembles smallpox. The 196,858-bp MPV genome was analyzed with regard to structural features and open reading frames. Each end of the genome contains an identical but oppositely oriented 6379-bp terminal inverted repetition, which similar to that of other orthopoxviruses, includes a putative telomere resolution sequence and short tandem repeats. Computer-assisted analysis was used to identify 190 open reading frames containing >/=60 amino acid residues. Of these, four were present within the inverted terminal repetition. MPV contained the known essential orthopoxvirus genes but only a subset of the putative immunomodulatory and host range genes. Sequence comparisons confirmed the assignment of MPV as a distinct species of orthopoxvirus that is not a direct ancestor or a direct descendent of variola virus, the causative agent of smallpox. PMID- 12083819 TI - Temporal acceleration of the human papillomavirus life cycle by adeno-associated virus (AAV) type 2 superinfection in natural host tissue. AB - Epidemiologically, certain human papillomaviruses are positively associated with cervical cancer, while adeno-associated viruses (AAV-2) are negatively associated with this same cancer. Both HPV and AAV productively replicate in differentiating keratinocytes of the skin and interact with each other. However, AAV has a relatively fast life cycle, generating infectious progeny by the third to fourth day of an organotypic epithelial raft culture. In contrast, HPV is slow, generating infectious progeny only after 10-12 days. As earlier studies indicated that these two skin-tropic virus types significantly affect each other's life cycle, we investigated if the temporal kinetics of the slow HPV life cycle was affected by the fast AAV in raft cultures. Here it is shown that the presence of AAV-2 at a variety of multiplicities of infection (m.o.i.) resulted in early onset HPV-31b DNA replication. Using plasmids which each expressed only one of the four rep proteins, an enhancement affect was seen for all four rep proteins of AAV, with Rep40 having the highest activity. Furthermore, AAV (m.o.i. of 5) also resulted in a temporally accelerated production of HPV infectious units, seen as early as Day 4, with high levels of viral progeny being produced by Day 6.5. Like earlier studies at Day 12, histological differences were seen at Day 6.5 between AAV-infected and mock-infected HPV/rafts. These data suggest that under specific conditions the AAV rep trans-factors can positively regulate HPV gene expression in addition to the usual negative regulation that has been consistently observed by the rep proteins. These data also suggest that AAV has a significant effect upon the temporal kinetics of the HPV life cycle in natural host tissue. However, it is unclear if or how this AAV-induced fast HPV life cycle mechanistically correlates with lower rates of HPV-associated cervical disease. PMID- 12083820 TI - Binding of adenovirus to its receptors in mouse astrocytes induces c-fos proto oncogene and apoptosis. AB - We have demonstrated that Ad.betaGal, a broadly used adenoviral vector of serotype 5, binds and induces proto-oncogene c-fos expression in quiescent cultures of mouse brain astrocytes. As observed in Northern blots, the expression of this immediate early gene is induced by viral infection in a dose-dependent manner, peaking at a multiplicity of infection (m.o.i.) of 100. The expression of c-fos is transient, being maximal after 30 min and disappearing 2 h after infection. A previously reported method was used to study the presence of receptors for adenovirus in the cellular membrane of murine astrocytes. After absorption of the virus, rabbit antibodies and 125I-protein A were used to form a sandwich on the cellular surface, and 9000 adenovirus-specific receptors were demonstrated on each astrocytic cell. Binding was temperature dependent and reached a plateau after 60 min. The specificity of c-fos induction is demonstrated by its neutralization by anti-adenovirus-specific antibodies. Although clear apoptosis cannot be demonstrated in vitro by DNA laddering, maybe due to a lack of sensitivity of the method, a statistically significant increase in caspase-3 activity is demonstrated in astrocyte cultures infected at a m.o.i. of 100 by adenovirus. Furthermore, a perfect colocalization is shown in vivo between cells infected with the Ad.betaGal vector and apoptotic astrocytes, as demonstrated by TdT-mediated dUTP nick end labeling (TUNEL) staining. The purpose of our study was to ascertain the potential for adenovirus as a gene therapy vector for neural disorders caused by astrocyte dysfunctions. PMID- 12083821 TI - Developmental expression of HERV-R (ERV3) and HERV-K in human tissue. AB - The human endogenous retroviruses (HERVs), ERV3 (HERV-R) and HERV-K, are both known to be transcriptionally active in human placenta. In the case of ERV3 there is also indirect evidence for its participation in cellular differentiation. In this study we examined the expression of ERV3 (HERV-R) and HERV-K in human normal fetal tissues by in situ hybridization. The highest level of ERV3 env expression was detected in primitive adrenal cortex. Elevated levels of expression were also found in the following developing tissues: kidneys (tubules), tongue, heart, liver, and central nervous system. Tissue-specific expression was found for HERV K rec (former cORF) but not for pol/int transcripts. The highest rec expression was found in placenta and levels slightly higher than sense control were found in the rest of the tissues examined. Pol/Int was not possible to quantitate. It appears that ERV3 is expressed in an organ-specific way during embryogenesis and might suggest a possible role in the development and differentiation of human tissues. PMID- 12083822 TI - Identification and genomic analysis of a second species of nucleopolyhedrovirus isolated from Mamestra configurata. AB - MacoNPV-96B is a nucleopolyhedrovirus isolated from naturally infected Mamestra configurata (Lepidoptera: Noctuidae) larvae. It was initially identified due to its completely different restriction endonuclease profile relative to the previously sequenced Mamestra configurata virus MacoNPV-90/2 (Q. Li, C. Donly, L. Li, L. G. Willis, D. A. Theilmann, and M. Erlandson, 2002, Virology 294, 106 121). The MacoNPV-96B host range and virulence were also found to differ significantly from those of the previous isolate. To further understand the complex of viruses infecting M. configurata, the genome of MacoNPV-96B was completely sequenced and analyzed in comparison with the genome of MacoNPV-90/2 and other sequenced baculoviruses. MacoNPV-96B consists of 158,482 bp, and 168 open reading frames (ORFs) of 150 nucleotides or longer with minimal overlap have been identified. The genome of MacoNPV-96B is 3422 bp larger than MacoNPV-90/2 and although gene arrangement is virtually identical, there are 9 ORFs unique to MacoNPV-96B and 10 unique to MacoNPV-90/2. bro genes were found to be associated with nonhomologous regions, suggesting that bro genes may facilitate recombination between genomes. A major difference in the gene content between the two viruses is a 5.4-kb insert in MacoNPV-96B, which is highly homologous to a cluster of Xestia c-nigrum granulovirus (XecnGV) ORFs, suggesting recent recombination events between these two viruses. Nucleotide sequence and amino acid sequence identity between the common ORFs of MacoNPV-96B and MacoNPV-90/2 average 87 and 90%, respectively. The sequence data suggest that MacoNPV-96B and MacoNPV-90/2 are closely related but have diverged and evolved into two separate species. This is the first study to identify highly related but separately evolving viruses in the same insect host and geographic location. A new Identity GeneParityPlot analysis was developed to perform a comparison of two viral genomes in gene content and arrangement as well as homology level of individual ORFs. PMID- 12083823 TI - Role of G protein beta3 subunit C825T and HLA class II polymorphisms in the immune response after HBV vaccination. AB - The G protein beta3 (GNB3) subunit and HLA are candidate genes predictive of immune response capacity. We therefore studied the influence of both gene systems on cellular and humoral immunity against hepatitis B virus (HBV) in 79 HBV booster-vaccinated healthy volunteers and an independent group of 77 probands after HBV basic immunization. Following booster vaccination, lymphocyte in vitro proliferation after stimulation with HBV surface antigen was 2.5-fold increased in GNB3 825T (TC + TT) vs CC allele carriers (P = 0.01) and was not influenced by HLA-DRB1 or DQB1 alleles. In addition, anti-HBs antibody titers in both groups were 2-fold increased in TC vs CC and decreased in TT vs CC allele carriers. However, antibody titers after HBV booster immunization were elevated in HLA DQB1*0301 carriers (P corrected = 0.027). In summary, the GNB3 825T allele appears as a marker particularly predictive of cellular and HLA-DQB1*0301 of humoral immune responses following HBV vaccination. PMID- 12083824 TI - In vivo effects of the Epstein-Barr virus small RNA EBER-1 on protein synthesis and cell growth regulation. AB - Recent studies have suggested a role for the Epstein-Barr virus-encoded RNA EBER 1 in malignant transformation. EBER-1 inhibits the activity of the protein kinase PKR, an inhibitor of protein synthesis with tumour suppressor properties. In human 293 cells and murine embryonic fibroblasts, transient expression of EBER-1 promoted total protein synthesis and enhanced the expression of cotransfected reporter genes. However reporter gene expression was stimulated equally well in cells from control and PKR knockout mice. NIH 3T3 cells stably expressing EBER-1 exhibited a greatly increased frequency of colony formation in soft agar, and protein synthesis in these cells was relatively resistant to inhibition by the calcium ionophore A23187. Nevertheless clones containing a high concentration of EBER-1 were not invariably tumourigenic. We conclude that EBER-1 can enhance protein synthesis by a PKR-independent mechanism and that, although this RNA may contribute to the oncogenic potential of Epstein-Barr virus, its expression is not always sufficient for malignant transformation. PMID- 12083826 TI - The herpesvirus saimiri tip484 and tip488 proteins both stimulate lck tyrosine protein kinase activity in vivo and in vitro. AB - Herpesvirus saimiri (HVS) of subgroup C efficiently induces leukemia in New World primates and transforms human lymphocytes. The viral tyrosine kinase interacting protein (Tip) binds to the tyrosine protein kinase Lck and is essential for transformation. Understanding how Tip modulates Lck activity is important for elucidating the mechanism of herpesvirus saimiri leukemogenesis. However, there are reports suggesting that whereas the Tip protein of HVS strain 484 stimulates the activity of Lck, the Tip protein of HVS strain 488 inhibits Lck. To determine whether these two divergent Tip proteins have opposite effects on Lck activity, we compared them in parallel. We found that both Tip proteins stimulated Lck kinase activity in vivo and in vitro and that both stimulated NF-AT- and STAT3 dependent transcription in T cells. Our data support the model that HVS infection increases the activity of Lck through the action of Tip. PMID- 12083825 TI - Human immunodeficiency virus type-1 activates lytic cycle replication of Kaposi's sarcoma-associated herpesvirus through induction of KSHV Rta. AB - Human immunodeficiency virus type-1 (HIV-1) infection dramatically increases the risk of development of Kaposi's sarcoma (KS) in individuals infected with Kaposi's sarcoma-associated herpesvirus (KSHV). In a primary effusion lymphoma (PEL) tissue culture model system, HIV-1 replication potently induced the lytic replication of KSHV and led to the secretion of soluble factors capable of inducing lytic KSHV replication in bystander cells. Here we demonstrate that HIV induces KSHV lytic replication through activation of the KSHV Rta. HIV gene expression activated the KSHV Rta promoter following viral infection or after transfection of proviral DNA. Although HIV-1 Tat has previously been implicated as an activator of KSHV lytic replication, Tat alone was unable to activate lytic replication and failed to activate the Rta promoter. We conclude that HIV activates KSHV lytic replication by inducing the KSHV Rta promoter and that factors other than HIV-1 Tat are required to mediate this effect. PMID- 12083827 TI - The nucleotide sequence of RNA1 of Lettuce big-vein virus, genus Varicosavirus, reveals its relation to nonsegmented negative-strand RNA viruses. AB - The complete nucleotide sequence of RNA1 from Lettuce big-vein virus (LBVV), the type member of the genus Varicosavirus, was determined. LBVV RNA1 consists of 6797 nucleotides and contains one large ORF that encodes a large (L) protein of 2040 amino acids with a predicted M(r) of 232,092. Northern blot hybridization analysis indicated that the LBVV RNA1 is a negative-sense RNA. Database searches showed that the amino acid sequence of L protein is homologous to those of L polymerases of nonsegmented negative-strand RNA viruses. A cluster dendrogram derived from alignments of the LBVV L protein and the L polymerases indicated that the L protein is most closely related to the L polymerases of plant rhabdoviruses. Transcription termination/polyadenylation signal-like poly(U) tracts that resemble those in rhabdovirus and paramyxovirus RNAs were present upstream and downstream of the coding region. Although LBVV is related to rhabdoviruses, a key distinguishing feature is that the genome of LBVV is segmented. The results reemphasize the need to reconsider the taxonomic position of varicosaviruses. PMID- 12083828 TI - Mutations in NS5B polymerase of hepatitis C virus: impacts on in vitro enzymatic activity and viral RNA replication in the subgenomic replicon cell culture. AB - Hepatitis C virus (HCV) nonstructural protein 5B (NS5B) is an RNA-dependent RNA polymerase (RdRp) essential for virus replication. Several consensus sequence motifs have been identified in NS5B, some of which have been shown to be critical for its enzymatic activity. A unique beta-hairpin structure located between amino acids 443 and 454 in the thumb subdomain has also been shown to play an important role in ensuring terminal initiation of RNA synthesis in vitro. However, the importance of these sequence and structural elements in viral RNA replication in infected cells has not been established, mainly due to the lack of a reliable cell culture system for HCV. In this study, we investigated the effect of several single amino acid substitutions and beta-hairpin truncations in NS5B on viral RNA replication by using the subgenomic replicon cell culture system. A strong correlation between in vitro polymerase activity and viral RNA replication was observed with most of the substitutions. Interestingly, truncations of the beta hairpin (by four and eight amino acid residues, respectively), which did not reduce the in vitro enzymatic activity, completely abolished the ability of the replicon RNA to replicate in Huh-7 cells, demonstrating its essential role in viral RNA replication. Furthermore, a conservative substitution in motif D, from an arginine residue (AMTR(345)), which is conserved among all HCV isolates, to a lysine residue, resulted in significant improvements in both transient RNA replication and colony formation efficiencies. This result also correlates with a previous observation that the enzymatic activity of NS5B increased by about 50% when the same NS5B substitution was introduced (V. Lohmann, F. Korner, U. Herian, and R. Bartenschlager, J. Virol. 1997, 71, 8416-8428). PMID- 12083829 TI - Hormone response element in SV40 late promoter directly affects synthesis of early as well as late viral RNAs. AB - We previously demonstrated that the presence of a hormone response element surrounding the transcription initiation site of the SV40 major late promoter (+1 HRE) confers a replication advantage to the virus in a cell-type-specific manner. We determine here the mechanism by which the +1 HRE confers this advantage by analyzing in detail the various stages of the viral life cycle of wild-type versus a +1 HRE mutant in MA-134 cells. We show that the mutant overexpresses late genes at the expense of early genes at early times after infection. This initial underproduction of early RNA leads, subsequently, to an underproduction of large T-antigen, viral DNA, and infectious virions. We conclude that the +1 HRE is necessary for the proper initial regulation of transcription from the early as well as late promoter so the cascade of subsequent events can be executed for the optimal production of virions. PMID- 12083830 TI - Respiratory syncytial virus immunobiology and pathogenesis. PMID- 12083832 TI - Induction of interferon-alpha and tumor necrosis factor-related apoptosis inducing ligand in human blood mononuclear cells by hemagglutinin-neuraminidase but not F protein of Newcastle disease virus. AB - To determine molecular viral components which can induce innate immune responses in human peripheral blood mononuclear cells (PBMC), we investigated the anti neoplastic agent Newcastle disease virus (NDV) and its two spike proteins hemagglutinin-neuraminidase (HN) and fusion protein (F). NDV was an excellent inducer in PBMC of IFN-alpha production and capable of inducing upregulation of plasma membrane expression of tumor necrosis factor related apoptosis inducing ligand (TRAIL). Viral replication was not required for these responses because NDV inactivated for 5 min by UV was as good as live NDV. NDV-modified and paraformaldehyde-fixed BHK cells could also trigger IFN-alpha and TRAIL induction, indicating that contacts of responder cells with NDV-modified cell surfaces are sufficient to induce these activities in PBMC. Antibodies against HN but not F were able to block these responses. Finally we could show that HN but not F induced IFN-alpha and TRAIL in PBMC. This was possible through the use of respective gene transfectants generated with the help of Semliki Forest virus (SFV) replicase-based DNA recombinant expression systems. Upon contact with BHK cells expressing HN but not F at their cell surface, human PBMC produced IFN alpha and some cells, including monocytes and T lymphocytes, upregulated cell surface TRAIL expression. PMID- 12083831 TI - Antigenic peptides of the epizootic hematopoietic necrosis virus. AB - The epizootic hematopoietic necrosis virus (EHNV) is a strain of the Iridovirus genus, which includes viruses seriously affecting native and aquacultured fish and amphibians. Despite its growing importance as a threat to fish farming, very little information is available on the biochemical and immunological nature of this virus. To identify and characterize the main antigenic determinants of EHNV, a panel of murine monoclonal antibodies was produced upon parenteral inoculation with live virus. A total of 124 primary hybridoma cultures from two fusions was found to produce antibodies reacting with EHNV by ELISA, but no neutralizing monoclonal antibody was detected. Twenty hybridoma cultures were randomly chosen for further study, and the antibodies secreted were analyzed by Western blotting, radioimmunoprecipitation, and immunostaining of infected cells. Only three MAbs immunoprecipitated the 50-kDa EHNV major capsid protein (MCP) from infected cell lysates, but they did not stain this protein in Western blotting. Eight and five further MAbs recognized peptides of approximately 15 and 18 kDa, respectively. Four antibodies could not be mapped into any viral protein, although they specifically immunostained virus-infected cells and reacted with purified EHNV virions by ELISA. These latter MAbs and the three antibodies directed at the MCP are likely to recognize conformation-dependent epitopes on the virus capsid proteins. PMID- 12083833 TI - Involvement of integrin alphavbeta3 in the pathogenesis of human immunodeficiency virus type 1 infection in monocytes. AB - Attachment of HIV to macrophages is a critical early event in the establishment of infection. In the present study, we demonstrate the involvement of integrin alphavbeta3 (vitronectin receptor) in HIV infection of peripheral blood monocyte derived macrophages. Culturing monocytes in the presence of M-CSF for 3 days upregulated expression of the alphav-containing integrins, alphavbeta3 and alphavbeta5. The increase in alphavbeta3 expression was accompanied by increased HIV-1 replication by monocytes. Immunoblot analysis showed that purified HIV gp120 protein interacted with CD4 and alphavbeta3 in immunoprecipitation experiments. Neutralizing antibodies against the alphavbeta3 integrin interfered with the coprecipitation of alphavbeta3 with an anti-gp120 antibody and substantially inhibited HIV infection of monocytes. Neutralizing antibodies against alphavbeta5 or beta1 integrins did not significantly affect HIV infection. These results indicate that HIV infection of primary monocytes requires differentiation of these cells and may involve alphavbeta3 interaction with the HIV-1 envelope protein gp120 for productive infection. PMID- 12083834 TI - Characterization of Bombyx mori nucleopolyhedrovirus orf68 gene that encodes a novel structural protein of budded virus. AB - All lepidopteran baculovirus genomes sequenced to date encode a homolog of the Bombyx mori nucleopolyhedrovirus (BmNPV) orf68 gene, suggesting that it performs an important role in the virus life cycle. In this article we describe the characterization of BmNPV orf68 gene. Northern and Western analyses demonstrated that orf68 gene was expressed as a late gene and encoded a structural protein of budded virus (BV). Immunohistochemical analysis by confocal microscopy showed that ORF68 protein was localized mainly in the nucleus of infected cells. To examine the function of orf68 gene, we constructed orf68 deletion mutant (BmD68) and characterized it in BmN cells and larvae of B. mori. BV production was delayed in BmD68-infected cells. The larval bioassays also demonstrated that deletion of orf68 did not reduce the infectivity, but mutant virus took 70 h longer to kill the host than wild-type BmNPV. In addition, dot-blot analysis showed viral DNA accumulated more slowly in mutant infected cells. Further examination suggested that BmD68 was less efficient in entry and budding from cells, although it seemed to possess normal attachment ability. These results suggest that ORF68 is a BV-associated protein involved in secondary infection from cell-to-cell. PMID- 12083835 TI - The cytoskeleton and the secretory pathway are not involved in targeting the cowpea mosaic virus movement protein to the cell periphery. AB - The movement protein (MP) of cowpea mosaic virus (CPMV) forms tubules on infected protoplasts and through plasmodesmata in infected plants. In protoplasts the MP fused to GFP (MP-GFP) was shown to localize in peripheral punctate structures and in long tubular structures extending from the protoplast surface. Using cytoskeletal assembly inhibitors (latrunculin B and oryzalin) and an inhibitor of the secretory pathway (brefeldin A), targeting of the MP to the peripheral punctate structures was demonstrated not to be dependent on an intact cytoskeleton or functional secretion pathway. Furthermore it was shown that a disrupted cytoskeleton had no effect on tubule formation but that the addition of brefeldin A severely inhibited tubule formation. The results presented in this paper suggest a role for a plasma membrane host factor in tubule formation of plant viral MPs. PMID- 12083837 TI - Target-cell-derived tRNA-like primers for reverse transcription support retroviral infection at low efficiency. AB - Reverse transcription of a retroviral genome takes place in the cytoplasm of an infected cell by a process primed by a producer-cell-derived tRNA annealed to an 18-nucleotide primer-binding site (PBS). By an assay involving primer complementation of PBS-mutated vectors we analyzed whether tRNA primers derived from the target cell can sustain reverse transcription during murine leukemia virus (MLV) infection. Transduction efficiencies were 4-5 orders of magnitude below those of comparable producer-cell complementations. However, successful usage of a target-cell-derived tRNA primer was proven by cases of correction of single mismatches between Akv-MLV vectors and complementary tRNA primers toward the primer sequence in the integrated vector. Thus, target-cell-derived tRNA-like primers are able to initiate first-strand cDNA synthesis and plus-strand transfer leading to a complete provirus, suggesting that endogenous tRNAs from the infected cell may also have access to the intracellular viral complex at that step of the replication cycle. PMID- 12083836 TI - Simian immunodeficiency virus encephalitis: analysis of envelope sequences from individual brain multinucleated giant cells and tissue samples. AB - Simian immunodeficiency virus (SIV)-infected macaques develop an encephalitis (SIVE) that is pathologically virtually indistinguishable from that associated with HIV infection, with multinucleated giant cells (MNGCs) being the principal histopathological manifestation. To dissect SIV variants responsible for MNGC development, we examined the relationships between env sequences transcribed in individual MNGCs and those from genomic DNA of brain and spleen tissues. The brain-specific variant found in all brain clones was dominant among the clones from MNGCs, suggesting a role in the formation of giant cells. Furthermore, two additional minor groups of sequences were present in MNGCs. One group consisted of sequences closely related to those from spleen, indicating recent and probably multiple episodes of neuroinvasion. The second group represented clones similar or identical to the initial inoculum. The survival of archival sequences and their activation presumably by the fusion of productively and quiescently infected macrophages/microglia identify the central nervous system as a possible anatomical reservoir for latent infection. PMID- 12083838 TI - Primary isolated human brain microvascular endothelial cells express diverse HIV/SIV-associated chemokine coreceptors and DC-SIGN and L-SIGN. AB - Chemokines have received increasing attention due to their inhibitory activities on human immunodeficiency virus type-1 (HIV-1) and simian immunodeficiency virus (SIV) replication and the potential for chemokine receptors to assist in HIV 1/SIV entry into permissive cells. Besides CD4, which is the major receptor for HIV-1 and SIV, a number of chemokine receptors including but not limited to APJ, CCR3, CXCR4, and CCR5 may be coreceptors for HIV-1/SIV, not only in peripheral blood and lymphoid tissues but also in the central nervous system (CNS). The present studies reveal the lack of CD4, but the significant expression of various chemokine receptors, APJ, CCR3, CXCR4, and CCR5, plus C-type lectins DC-SIGN and L-SIGN on isolated primary human brain microvascular endothelial cells (MVECs). As these MVECs do not express CD4, this suggests a CD4-independent HIV/SIV entry/infection of these cells, which are the major cells constituting the human blood-brain barrier. We also found that chemokines for cognate chemokine receptors individually were unable to block binding of HIV-1 to brain MVECs. These results reveal that in primary isolated brain MVECs viral attachment is mediated by a possible previously unknown receptor(s) or by cooperative activity of various receptors. Moreover, mRNA transcripts for DC-SIGN/L-SIGN, as well as DC-SIGN protein expression, suggest the capability of MVECs to attach viral particles on cell surfaces, even though polyclonal antisera for DC-SIGN did not affect viral binding to these cells. These data will assist in further understanding lentiviral entry into the CNS. PMID- 12083839 TI - Cell-associated HIV-1 infection of salivary gland epithelial cell lines. AB - In this study we investigated the potential of cell-associated primary isolates of HIV-1 to infect epithelial cells of the oral cavity, using two cell lines originating in the salivary gland, HSY and HSG. Neither of these cell lines expresses surface CD4; however, both express the alternative epithelial receptor galactosylceramide and the coreceptor CXCR4. The ability of these cell lines to be infected via cell-associated infection was studied using a lab-adapted strain and primary isolates of HIV-1. Both HIV-1 phenotypes were able to infect these cell lines. The ligand for CXCR4 was able to block infection of both cell lines, whereas the ligands for CCR5 were able to block infection in HSY only, indicating that these receptors were utilized for entry into the epithelial cells. These findings demonstrate that epithelial cells of the oral cavity can be productively infected with HIV-1 by cell-borne virus in the absence of CD4. PMID- 12083840 TI - Stability of rice yellow mottle virus and cellular compartmentalization during the infection process in Oryza sativa (L.). AB - Rice yellow mottle virus (RYMV) is icosahedral in morphology and known to swell in vitro, but the biological function of swollen particles remains unknown. Anion exchange chromatography was used to identify three markedly stable forms of RYMV particles from infected plants: (1) an unstable swollen form lacking Ca2+ and dependent upon basic pH; (2) a more stable transitional form lacking Ca2+ but dependent upon acidic pH; and (3) a pH-independent, stable, compact form containing Ca2+. Particle stability increased over the time course of infection in rice plants: transitional and swollen forms were abundant during early infection (2 weeks postinfection), whereas compact forms increased during later stages of infection. Electron microscopy of infected tissue revealed virus particles in vacuoles of xylem parenchyma and mesophyll cells early in the time course of infection and suggested that vacuoles and other vesicles were the major storage compartments for virus particles. We propose a model in which virus maturation is associated with the virus accumulation in vacuoles. In this acidic compartment, virus particles may bind Ca2+ to produce a highly stable, compact form of the virus. The localization of subcellular RYMV isoforms in infected cells and the corresponding biological properties of the virus are discussed. PMID- 12083841 TI - Sequence analysis of an isolate from a fatal human infection of Australian bat lyssavirus. AB - Australian bat lyssavirus (ABLV), which occurs in pteropid and insectivorous bat populations, causes a rabies-like encephalitis in infected humans. We report the first complete sequence of an ABLV isolate obtained from a human who developed symptoms 27 months after being bitten by an infected flying fox. This isolate is the smallest lyssavirus to be sequenced, with a size of 11,918 nucleotides. Analyses of previously unsequenced regions and the complete genome confirm its close relationship with classical rabies viruses. In addition, a leucine zipper like motif, not present in the other lyssaviruses, was found in the conserved domain I of the polymerase protein. This is the first report of a lyssavirus to vary in an 11-nucleotide, strictly conserved, complementary terminal sequence. This region is thought to encode important cis-acting regulatory signals; ABLV variation indicates a greater degree of flexibility than was thought for lyssaviruses in this region. A comparison of the pteropid and insectivorous isolates of ABLV indicates considerable differences between the two viruses. If the divergence of the two occurred on the Australian mainland, ABLV may have been endemic to Australia well before European colonisation. PMID- 12083842 TI - Role of specific cell-mediated cytotoxicity in protecting fish from viral infections. AB - We report the in vivo role of specific cytotoxic cells in protecting fish from acute viral infections. Specifically, we found that (1) there is an inverse relationship between cytotoxic activities and viral load and (2) adoptive transfer of immune leukocytes prevented viral infection. Crucian carp hematopoietic necrosis virus (CHNV), which has a virulence to ginbuna crucian carp, was recently found and identified in Japan. Specific cell-mediated cytotoxicity of ginbuna leukocytes against CHNV-infected syngeneic cells was induced as a result of intraperitoneal inoculation with CHNV. This cytotoxicity was not induced against either virus-infected allogeneic cells or eel rhabidovirus- (EVA) infected syngeneic cells. In these respects, the cytotoxic activity was similar to that of mammalian cytotoxic T-lymphocyte (CTL) activity. Viral titers of tissues from infected fish were remarkably reduced 8 days after infections, when specific cytotoxic activity reached a peak. This result suggested that specific cytotoxic cells were responsible for the early control of CHNV replication. On the other hand, CHNV-specific antibody was greatly increased when the virus was eliminated by cytotoxic activities. The effectiveness of the virus-specific cytotoxicity was evaluated using adoptive cell transfer. Recipients that received leukocytes from immune syngeneic donors escaped CHNV infection. These findings suggest that virus-specific cytotoxic cells have a role in controlling viral infections in a fish. PMID- 12083843 TI - Characterization of the equilibrium between the native and fusion-inactive conformation of rabies virus glycoprotein indicates that the fusion complex is made of several trimers. AB - Rabies virus-induced membrane fusion is triggered at low pH and is mediated by the trimeric viral glycoprotein (G). G assumes three conformations: the native state (N) detected above pH 7; the activated state (A), which initiates the fusion process; and the fusion-inactive conformation (I) observed after prolonged incubation at low pH. Differently from other viral fusogenic glycoproteins, G in the I state recovers its native conformation when reincubated above pH 7. Here, we demonstrate that there is a thermodynamic equilibrium between the different states of G between pH 6 and pH 7.5. The study of this equilibrium at various pH values indicated that the conformational change toward I is induced by the protonation of at least three residues per trimer. Finally, studies on the mechanism leading to low pH induced fusion inactivation indicated that a large number of G molecules is required for stable hydrophobic interaction of the virus with the target membrane. PMID- 12083844 TI - Evaluation of the replication and immunogenicity of recombinant human parainfluenza virus type 3 vectors expressing up to three foreign glycoproteins. AB - The level of replication and immunogenicity of recombinant parainfluenza virus type 3 (rHPIV3) bearing one, two, or three gene insertions expressing foreign protective antigens was examined. cDNA-derived recombinant HPIV3s bearing genes encoding the open reading frames (ORFs) of the hemagglutinin-neuraminidase (HN) of HPIV1, the HN of HPIV2, or the hemagglutinin (HA) of measles virus replicated efficiently in vitro, including the largest recombinant, which had three gene unit insertions and which was almost 23 kb in length, 50% longer than unmodified HPIV3. Several viruses were recovered from cDNAs whose genome length was not a multiple of six nucleotides and these contained nucleotide insertions that corrected the length to be a multiple of 6, confirming that the "rule of six" applies to HPIV3. Using a hemagglutination inhibition assay, we determined that the HPIV1 HN expressed by recombinant HPIV3 was incorporated into HPIV3 virions, whereas using this assay incorporation of the HPIV2 HN could not be detected. HPIV3 virions bearing HPIV1 HN were not neutralized by HPIV1 antiserum but were readily neutralized by antibodies to the HPIV3 HN or fusion protein (F). Viruses with inserts were restricted for replication in the respiratory tract of hamsters, and the level of restriction was a function of the total number of genes inserted, the nature of the insert, and the position of the inserted gene in the gene order. A single insert of HPIV2 HN or measles virus HA reduced the in vivo replication of rHPIV3 up to 25-fold, whereas the HPIV1 HN insert decreased replication almost 1000-fold. This indicates that the HPIV1 HN insert has an attenuating effect in addition to that of the extra gene insert itself, presumably because it is incorporated into the virus particle. Viruses containing two inserts were generally more attenuated than those with a single insert, and viruses with three inserts were over-attenuated for replication in hamsters. Inserts between the N and P genes were slightly more attenuating than those between the P and the M genes. A recombinant HPIV3 bearing both the HPIV1 and the HPIV2 HN genes (r1HN 2HN) was attenuated, immunogenic, and protected immunized hamsters from challenge with HPIV1, HPIV2, and HPIV3. Thus, it is possible to use a single HPIV vector expressing two foreign gene inserts to protect infants and young children from the severe lower respiratory tract disease caused by the three major human PIV pathogens. PMID- 12083845 TI - Sendai virus, a murine parainfluenza virus type 1, replicates to a level similar to human PIV1 in the upper and lower respiratory tract of African green monkeys and chimpanzees. AB - Human parainfluenza virus type 1 (HPIV1), a major cause of croup in infants and young children, accounts for 6% of hospitalizations for pediatric respiratory tract disease. The antigenically related Sendai virus, referred to here as murine PIV1 (MPIV1), is being considered for use as a live-attenuated vaccine to protect against HPIV1 (J. L. Hurwitz, K. F. Soike, M. Y., Sangster, A. Portner, R. E. Sealy, D. H. Dawson, and C. Coleclough, 1997, Vaccine 15(5), 533-540) and also as a recombinant vaccine vector expressing antigens to protect against viral disease in humans. However, in the 1950s MPIV1 was reported to have been isolated from humans, suggesting that zoonotic transmission might have occurred. It is therefore important to examine the ability of MPIV1 to replicate in nonhuman primates, i.e., surrogate hosts for humans. In the present study the level of replication of MPIV1 and HPIV1 was compared in African green monkeys and chimpanzees. Surprisingly, MPIV1 replicated as efficiently as HPIV1 in the upper and lower respiratory tract of African green monkeys at doses of 10(4) and 10(6) and replicated only slightly less efficiently at both sites in chimpanzees. African green monkeys immunized with MPIV1 were highly resistant to subsequent challenge with HPIV1 even though MPIV1 did not induce a detectable HPIV1 neutralizing antibody response. The high level of replication of MPIV1 observed in the upper and lower respiratory tract of these primates suggests that MPIV1 likely would require significant attenuation before it could be given to humans as a vaccine against HPIV1 or as a vaccine vector. Its ability to efficiently replicate in nonhuman primates suggests that MPIV1 lacks a significant host range restriction in primates and could theoretically cause zoonotic disease in humans. PMID- 12083846 TI - Cell-matrix interactions in cancer spreading--effect of aging. AB - Passage of connective tissue barriers is a compulsory step in cancer spreading. This process involves a series of steps involving cell-matrix interactions. The complexity of these steps increased considerably during the last decades as a result of the identification of a large number of newly discovered macromolecular components of the extracellular matrix (ECM). It was also recognized that malignant cells can and do modify the production as well as the degradation of ECM components. The rapid development of aging biology during the second-half of the last century also contributed to the complexity but also to the importance of the newly emerging picture of cancer progression as a function of age. The structure and composition of the ECM is strongly age-dependent and may well represent one of the important factors influencing the age-distribution of human malignancies. As will be evident from the following reviews this relatively new and rapidly increasing picture of the age-dependence of cell-matrix interactions and cancer spreading should trigger more intense efforts on this relatively new discipline in cancer research. PMID- 12083847 TI - The biology of conformation in the regulation of the senescent and transformed cell phenotypes. AB - The cytoskeleton and the composition of the cytoplasmic membrane of normal somatic cells are modified during proliferation in vitro. The loss of the proliferative potential during serial divisions is due in part to these structural modifications that induce a decline in the cell conformational flexibility. During viral transformation, the changes in the affinity of the cell to its matrix and to neighboring cells increase the cell migratory capability maintaining the conformational flexibility; this way the cells can proliferate to densities where normal cells stop dividing. Cell proliferation, the transformed phenotype, and differentiation could be modulated by changing the electric charge of a substratum. Results support the view that the biology of conformation is crucial for the expression of these cell properties. PMID- 12083848 TI - Proteoglycans and tumor progression: Janus-faced molecules with contradictory functions in cancer. AB - Understanding the details of the molecular mechanism of tumor dissemination revealed that several proteoglycan species are involved in the process but their role can be described as Janus-faced. One level of proteoglycan alterations is at the expression of their genes coding for the core protein. Characteristically, in progressing tumors two patterns emerged: loss or neoexpression of surface proteoglycans (PG) depending on the initial expression pattern of the cell type of origin. The situation is similarly complex concerning the changes of glycosaminoglycan (GAG) of the PG during tumor progression. This is due to the fact that the majority of PGs involved is hybrid molecule meaning that their core protein can be glycanated both with chondroitin and heparan sulfate. However, such an alteration in glycanation of PG may fundamentally change the function of the molecule, especially the one operating at the cell surface. Among the extracellular PGs, decorin emerged as inhibitor of progression while perlecan as a promoter of the process. Analysis of the available data indicate that during metastatization tumor cells must express at least one cell surface HSPG species from the syndecan-glypican-CD44v3 group. Furthermore, the HS-chain of these proteoglycan(s) carry important molecular signatures (suphution or epimerization patterns). Experimental data suggest that tumor cell surface heparan sulfate (PG) may provide a target for specific anti-metastatic interventions. PMID- 12083849 TI - Fibronectin in malignancy. AB - Fibronectin (Fn) was the first 'structural' glycoprotein intensively studied as an ubiquitous matrix component of early phylogenetic appearance. Its age dependent increase in plasma and tissues may be accompanied in pathological states, especially in tumor growth, by its proteolytic breakdown by a number of neutral proteases. It was also shown that several of its proteolytic breakdown products exhibit unexpected and mostly harmful biological activities. The first of these effects was a potentiation of malignant transformation. Some fragments had proteolytic activity, others behaved as proinflammatory agents stimulating IL 1 and collagenolytic MMP up-regulation. This matricryptic potential of Fn was followed by several other examples of proteolytic production of biologically active peptides. The study of solid human tumors showed among the early signs of malignant transformation the fragmentation of pericellular Fn, concommitent with the increase of its production by the peritumoral stroma. These results should encourage further investigations concerning the potential importance of Fn production and breakdown during cancer progression. PMID- 12083850 TI - Laminin isoforms in tumor invasion, angiogenesis and metastasis. AB - Laminins are a growing family of alphabetagamma heterotrimeric proteins, commonly found in basement membranes (BMs). These large molecules promote cell adhesion and migration via integrins and other cell-surface receptors. Over 12 laminin isoforms are presently known. The various isoforms have a cell- and tissue specific expression and are differentially recognized by integrins. Expression of laminin isoforms in tumors usually reflects expression in their normal counterparts. However, during tumor invasion, loss of the BM barrier occurs and a discontinuous pattern of laminin staining is observed. In carcinomas, tumor cells at the invading front strongly express intracellularly the gamma2 chain, a component of laminin-5. Remodeling of the vascular BM is observed during angiogenesis, and penetration of several BMs occurs during tumor dissemination and metastasis. Thus, disregulated cell-laminin interactions are major traits of malignant disorders. PMID- 12083851 TI - Role of elastin-matrix interactions in tumor progression. AB - Data from the literature now indicate that cancer cells can specifically interact with the unique extracellular matrix protein, elastin. The interaction is mediated by two elastin-binding proteins (EBP), S-gal/EBP (organized into the elasin receptor/elastonectin complex) and galectin-3, components of two laminin receptors. Studies revealed that the expression of both EBPs is closely associated to the invasive/metastatic potential of various cancer types. This is due to the fact that elastin-ligation of S-gal/EBP induces motogenic, as well as mitogenic signals and releases various elastases from cancer cells and the induction depends on the metastatic potential. Studies also demonstrated that certain cancer cells can synthesize elastin and express lysyl oxydase, providing explanation for frequent appearance of elastic tissue in tumors such as breast or gastric cancers. Clinico-pathological data suggest some correlation with tumor progression of the presence of the elastic tumor stroma. Since elastic tissue may be a significant reservoir of angiostatic molecule(s) this extracellular matrix protein can also have a role in tumor-induced angiogenesis. Soluble elastin as well as elastin peptides are potent inhibitors of the metastatic process in experimental tumor models. On the other hand, elastin peptides can also be used to design targeted therapies exploiting the unique physicochemical nature of this matrix protein. Altogether, these data suggest a significant role for tumor cell elastin interactions in tumor progression. PMID- 12083852 TI - Putative role of 67 kDa elastin-laminin receptor in tumor invasion. AB - Cellular regulatory mechanisms normally maintain a delicate balance between cell proliferation, quiescence and death. The imbalance between these functions resulting from molecular intracellular changes is a key factor in tumorigenesis. Tumor cells detaching from the primary tumor possess a propension for invasion and metastasis formation. These tumor cells can attach, migrate, proliferate and grow in host tissue. The surrounding extracellular matrix (ECM) modulates these functions. It is now widely accepted that cell-matrix interactions play an important role in these processes. Most investigators concentrated their attention on the role of integrins in the above processes. There are, however, only scant data on the role of elastin and its receptors in tumor invasion. Nevertheless, experimental evidence indicates that the 67 kDa elastin-laminin receptor (ELR) subunit plays an important role in tumor invasion by mediating essential tumor cell functions leading to metastases. In this review we will concentrate on the putative role of the 67 kDa ELR subunit in tumor invasion. PMID- 12083853 TI - Matrix-directed regulation of pericellular proteolysis and tumor progression. AB - The microenvironment of cancer cells, composed of extracellular matrix (ECM) macromolecules, plays a pivotal function in tumor progression. ECM preexisting modules or cryptic sites revealed by partial enzymatic hydrolysis positively or negatively regulate matrix metalloproteinase (MMP) expression and activation, further influencing matrix invasion by cancer cells. Pericellular activation of gelatinase A (MMP-2) proceeds via the formation of a complex involving its inhibitor, TIMP-2, its activator(s), MT-MMPs and alphavbeta3 integrin forming a docking system. This proteinase has been invariably linked to cancer cell invasive potential and is often predictive of a poor survival. MMP-2 degrades most ECM macromolecules and appears to act as a main 'decryptase'. ECM modulation of MMP-2 activation pathway thus influences angiogenesis and tumor growth. For instance the noncollagenous domain of alpha3 chain of type IV collagen, through alphavbeta3 integrin binding, inhibits both MT1-MMP and alphavbeta3 integrin expression from melanoma cells and empedes cell migration and proliferation. At the opposite, a particular module in elastin (VGVAPG) with type VIII beta turn conformation stimulates MT1-MMP and proMMP-2 activation through binding to S-gal elastin receptor, and increases the matrix invasive capacity of several cancer cell lines and endothelial cells. Endocytosis emerges as a main mechanism controlling MMP-2, and also other MMPs; it proceeds via the formation of a MMP thrombospondin(s) complex further recognized by the LRP scavenger receptor. ECM undergoes conspicuous variations with aging linked to alterations of tissue organization and post-translational modifications of matrix constituents that modify cell-matrix interactions and MMP-2 activation pathway. PMID- 12083854 TI - Extracellular matrix components in breast carcinomas. AB - A malignant process interferes with the normal 'programme' of extracellular matrix biosynthesis and can modify extensively the structure and composition of the matrix. This effect appears to be attributable to several processes such as direct production of some selected matrix macromolecules by malignant cells or indirectly by the production of factors by malignant cells interfering with the regulation of normal matrix production. Other possibilities may also exist, such as the direct action of an environmental carcinogen on otherwise normal mesenchymal cells. The result is a more or less profound modification of tissue structure and composition with possible feedback effects on the malignant process. Some examples will be discussed such as elastin production by some tumours as well as the biosynthesis of some other selected matrix macromolecules as tenascin and osteopontin by breast tumours. Although the detailed mechanisms of these specific matrix productions is not yet completely elucidated, the rapidly increasing knowledge on the regulation of specific matrix production process and deranged matrix production might represent a new area of crosstalk between cancer research and matrix biology. PMID- 12083855 TI - Myoglobin-induced lipid oxidation. A review. AB - An overview of myoglobin-initiated lipid oxidation in simple model systems, muscle, and muscle-based foods is presented. The potential role of myoglobin spin and redox states in initiating lipid oxidation is reviewed. Proposed mechanisms for myoglobin-initiated lipid oxidation in muscle tissue (pH 7.4) and meat (pH 5.5) are evaluated with the purpose of putting forward general mechanisms explaining present observations regarding the catalytic events. PMID- 12083856 TI - Detection of hazelnut oil adulteration using FT-IR spectroscopy. AB - Fourier transform infrared spectroscopy (FT-IR) was used to detect the adulteration of hazelnut oil with different types of oils and to detect the adulteration of extra-virgin olive oil with hazelnut oil. Spectra of hazelnut oil, seven other types of oils, extra-virgin olive oil, and the adulterated oils were collected with a FT-IR equipped with a ZnSe-ATR accessory and a MCTA detector. Discriminant analysis and partial least-squares analysis were used to analyze the data. Classification of hazelnut oil, olive oil, and the other types of oils was achieved successfully with FT-IR. The detection level for sunflower oil adulteration of hazelnut oil was 2%, and the correlation coefficient for the PLS model was 0.99. Adulteration of virgin olive oil with hazelnut oil could be detected only at levels of 25% and higher. PMID- 12083857 TI - Near-infrared reflectance spectroscopy (NIRS) enables the fast and accurate prediction of essential amino acid contents. 2. Results for wheat, barley, corn, triticale, wheat bran/middlings, rice bran, and sorghum. AB - Further NIRS calibrations were developed for the accurate and fast prediction of the total contents of methionine, cystine, lysine, threonine, tryptophan, and other essential amino acids, protein, and moisture in the most important cereals and brans or middlings for animal feed production. More than 1100 samples of global origin collected over five years were analyzed for amino acids following the Official Methods of the United States and European Union. Detailed data and graphics are given to characterize the obtained calibration equations. NIRS was validated with 98 independent samples for wheat and 78 samples for corn and compared to amino acid predictions using linear crude protein regression equations. With a few exceptions, validation showed that 70-98% of the amino acid variance in the samples could be explained using NIRS. Especially for lysine and methionine, the most limiting amino acids for farm animals, NIRS can predict contents in cereals much better than crude protein regressions. Through low cost and high speed of analysis NIRS enables the amino acid analysis of many samples in order to improve the accuracy of feed formulation and obtain better quality and lower production costs. PMID- 12083859 TI - Determination of angiotensin-converting enzyme inhibitory peptide Leu-Lys-Pro-Asn Met (LKPNM) in bonito muscle hydrolysates by LC-MS/MS. AB - Proteolytic digestion of dried bonito muscle with thermolysin produces a hydrolysate with strong angiotensin-converting enzyme (ACE) inhibitory activity and is the basis of a dietary supplement with antihypertensive activity. A major portion of the ACE activity was shown previously to arise from the peptide Leu Lys-Pro-Asn-Met (LKPNM). A straightforward method to quantify this peptide was developed using one-step C18 solid-phase extraction (SPE) followed by LC-MS/MS quantification. The SPE step resulted in a hydrolysate that was still crude, as illustrated by combined size-exclusion chromatography/multi-angle laser light scattering detection that showed that a major fraction of oligopeptides were in the 2-20 kDa range. This fraction has a weight-average molecular weight (M(w)) of approximately 5.0 kDa. Method validation for specificity, linearity, accuracy, precision, and reproducibility showed that standard additions of synthetic LKPNM to bonito extract with SPE enrichment followed by LC-MS/MS is a suitably robust procedure for the determination of LKPNM content. The method was also successful for encapsulated powders in which the excipients used are insoluble in water and could be removed by centrifugation. PMID- 12083858 TI - Characterization of irradiated starches by using FT-Raman and FTIR spectroscopy. AB - Fourier transform infrared (FTIR) and Fourier transform Raman (FT-Raman) methods were used for rapid characterization and classification of selected irradiated starch samples. Biochemical changes due to irradiation were detected using the two vibrational spectroscopic techniques, and canonical variate analysis (CVA) was applied to the spectral data for discriminating starch samples based on the extent of irradiation. The O-H (3000-3600 cm(-1)) stretch, C-H (2800-3000 cm(-1)) stretch, the skeletal mode vibration of the glycosidic linkage (900-950 cm(-1)) in both Raman and infrared spectra, and the infrared band of water adsorbed in the amorphous parts of starches (1550-1750 cm(-1)) were employed in classification analysis of irradiated starches. Spectral data related to water adsorbed in the noncrystalline regions of starches provided a better classification of irradiated starches with 5 partial least-squares (PLS) factors in the multivariate model. PMID- 12083860 TI - Determination of mesotrione residues and metabolites in crops, soil, and water by liquid chromatography with fluorescence detection. AB - A method for the determination of residues of mesotrione and two metabolites in a variety of environmental matrixes has been developed. Mesotrione, a new selective herbicide for use in corn, is 2-(4-methylsulfonyl-2-nitrobenzoyl)-1,3 cyclohexanedione. The metabolite 4-methylsulfonyl-2-nitrobenzoic acid (MNBA) is determined with the parent compound in crops, whereas two metabolites, 2-amino-4 methylsulfonyl-benzoic acid (AMBA) and MNBA are determined with parent in soil and water. Crop samples are macerated with an acetonitrile/water mixture, and an aliquot is evaporated and acidified then centrifuged. Soil is shaken with an ammonium hydroxide solution, and an aliquot is acidified then centrifuged. For water analysis, an aliquot is acidified. Crop and soil extracts, and water, are cleaned up using reverse-phase high-performance liquid chromatography (RPHPLC) with mesotrione and MNBA isolated using a fraction collector. During this clean up, AMBA is determined in soil and water samples using fluorescence detection. The collected mesotrione and MNBA fractions are converted into AMBA via oxidation followed by reduction in the case of mesotrione, or by reduction alone in the case of MNBA. Both fractions are analyzed by RPHPLC with fluorescence detection using an AMBA external reference standard. The method was tested on corn grain, fodder, and forage, as well as on sugar cane. The limits of quantitation (LOQ) for each analyte are 0.01 mg/kg for crops, 0.005 mg/kg for soil, and 0.10 microg/L for water. Method fortification recoveries from all crop commodities averaged 79% (CV = 7%, n = 37 and 82% (CV = 5%, n = 37) for mesotrione and MNBA, respectively. Soil was fortified at 0.005 and 0.05 mg/kg. Recoveries were 79% (CV = 4%, n = 12), 96% (CV = 2%, n = 12), and 89% (CV = 2%, n = 12) for mesotrione, MNBA, and AMBA, respectively. Groundwater, drinking water, seawater, and river water were fortified at 0.1 and 1.0 microg/L. Recoveries for all waters were 80% (CV = 7%, n = 51), 94% (CV = 4%, n = 52), and 93% (CV = 9%, n = 51) for mesotrione, MNBA, and AMBA, respectively. PMID- 12083861 TI - 2H/(1)H and (13)C/(12)C isotope ratios of trans-anethole using gas chromatography isotope ratio mass spectrometry. AB - Authenticity assessment of trans-anethole is deduced from (2)H/(1)H and (13)C/(12)C isotope ratios, determined by gas chromatography-isotope ratio mass spectrometry (GC-IRMS). For that purpose, self-prepared anise and fennel oils, and synthetic and "natural" samples of trans-anethole, as well as commercially available anise and fennel oils have been investigated. Authenticity ranges of (2)H/(1)H and (13)C/(12)C isotope ratios of trans-anethole were defined. Scope and limitations of the applied online GC-IRMS techniques are discussed. PMID- 12083862 TI - Selenium accumulation in beef: effect of dietary selenium and geographical area of animal origin. AB - Selenium (Se) is an essential nutrient with multiple human health benefits; the single most important dietary source of Se is beef. The Se content of beef varies, and cattle fed a high selenium diet may have Se concentrations in beef that are well above average. Such beef is potentially a unique supplemental source of dietary Se. To examine factors affecting Se accumulation in beef, 16 steers (initial wt 374.4 +/- 33.7 kg) were taken from seleniferous or nonseleniferous areas and fed in a 2 x 2 factorial design with diets high or moderate in Se (11.9 or 0.62 mg Se/kg diet). Diets contained 50% alfalfa, 25% wheat, and 25% corn on a dry matter basis. All dietary Se was from agricultural products, and Se in the high Se diet was primarily from high Se wheat and alfalfa hay. A loin muscle biopsy was taken at the start of the trial to determine initial Se content of beef. Steers were slaughtered after 14 weeks of the trial, and edible carcass (round, sirloin, shoulder clod, and ribeye) and organ samples were collected. Diets did not affect growth or feed intake (P > 0.05), and Se toxicity signs were not observed. Different cuts of meat had similar Se concentrations, and the Se content of all cuts was increased by both high dietary Se and high Se background. Except for liver and kidney, Se in tissues was increased by seleniferous background (P < 0.02) and high dietary Se (P < 0.001). Kidney Se concentrations of animals fed the high Se diet were lowest in animals from seleniferous areas (P = 0.04), suggesting a possible adaptation to the high Se diet. These results demonstrate that cattle fed diets high in Se from agricultural products will accumulate substantial amounts of Se in the beef without developing signs of Se toxicity and that prior Se status regulates Se accumulation in some organs. They further demonstrate that management practices may be altered so as to make beef a significant source of dietary Se. PMID- 12083863 TI - Antimicrobial screening of Mentha piperita essential oils. AB - Essential oils of peppermint Mentha piperita L. (Lamiaceae), which are used in flavors, fragrances, and pharmaceuticals, were investigated for their antimicrobial properties against 21 human and plant pathogenic microorganisms. The bioactivity of the oils menthol and menthone was compared using the combination of in vitro techniques such as microdilution, agar diffusion, and bioautography. It was shown that all of the peppermint oils screened strongly inhibited plant pathogenic microorganisms, whereas human pathogens were only moderately inhibited. Chemical compositions of the oils were analyzed by GC and GC/MS. Using the bioautography assay, menthol was found to be responsible for the antimicrobial activity of these oils. PMID- 12083864 TI - Supercritical fluid extraction of alkylamides from Echinacea angustifolia. AB - Echinacea has been known for its immunostimulatory activity, and its alkylamide components have been linked to such biological activity. Consequently, alkylamides in Echinacea angustifolia were extracted using supercritical carbon dioxide from fresh and dried roots at 45-60 degrees C and 34-55 MPa, and the alkylamide yield in the extracts was determined. The yield of alkylamides from fresh roots increased with temperature yet decreased with pressure, whereas the yield from air-dried roots (moisture content 8.4%) increased with both temperature and pressure. Freeze-drying of the roots to a moisture content of 4.9% did not result in any further increase in the yield compared to that of air dried roots. Alkylamide yield of the ground dried roots extract was the highest (p < or = 0.05) among those from fresh, ground and unground E. angustifolia roots. Supercritical fluid extraction therefore shows potential for the recovery of alkylamides from dried Echinacea roots. PMID- 12083866 TI - Relationship between color (instrumental and visual) and chlorophyll contents in soybean seeds during ripening. AB - The correlation between chlorophyll content and quantitative color parameters was investigated in order to find an indirect method for predicting green pigment in ripening soybean seeds. Five Brazilian soybean varieties harvested at different maturity stages (R(6) to R(8) according to the scale of Fehr & Caviness) and dried under two conditions (in oven at 40 degrees C with circulating air and at ambient temperature around 25 degrees C) were analyzed in two consecutive years. The slow-dried seeds at 25 degrees C lost chlorophyll faster, whereas drying at 40 degrees C did not result in yellowing of seeds. High and significant linear correlations between a value and total chlorophyll were obtained over the whole maturation period and on both conditions of drying. From an industrial point of view it appears that a value, obtained by the CIE-L*a*b* method, seems to be a good tool to be applied for quality control and classifying soybean seeds for different purposes. PMID- 12083865 TI - Effects of tea components on the response of GABA(A) receptors expressed in Xenopus Oocytes. AB - To study the effects of tea components on ionotropic gamma-aminobutyric acid (GABA) receptor response, ionotropic GABA receptors (GABA(A) receptors) were expressed in Xenopus oocytes by injecting cRNAs synthesized from cloned cDNAs of the alpha(1) and beta(1) subunits of the bovine receptors, and their electrical responses were measured by a voltage clamping method. Extracts of green tea, black tea, and oolong tea in an aqueous solution induced the GABA-elicited response, which showed that these teas contain GABA, whereas coffee does not. Caffeine weakly inhibited the response in a competitive manner (K(i) = 15 mM), and (+)-catechin inhibited it in a noncompetitive one (K(i) = 1.7 mM). Especially, two catechin derivatives, (-)-epicatechin gallate and (-) epigallocatechin gallate, inhibited the response strongly. Alcohols such as leaf alcohol or linalool potentiated the response, possibly because their binding to the potentiation site enhances the GABA-binding affinity to GABA(A) receptors when they bind. Extracts of green tea made with ethyl ether, which must contain lipophilic components of green tea, inhibited the response elicited by GABA, possibly because the amounts of caffeine and catechin derivatives were much larger than fragrant alcohols in such extracts of tea. PMID- 12083867 TI - Stabilization of caffeic acid derivatives in Echinacea purpurea L. glycerin extract. AB - Recent work has shown that enzymatic degradation and oxidation of cichoric acid and other caffeic derivatives occurs in Echinacea preparations. However, very little is known as to the means of stabilizing these phytopreparations. To stabilize the glycerin extract of Echinacea purpurea, we have evaluated the effects of 3 natural antioxidants (citric acid, malic acid, and hibiscus extract) on the stability of the major caffeic acid derivatives (caftaric acid, caffeic acid, cichoric acid, and 2-O-feruloyl-tartaric acid). Chlorogenic acid, which normally occurs in an ethanol extract of E. purpurea, was not present in the glycerin extract. The caffeic acid derivatives, with the exception of 2-O feruloyl-tartaric acid, were subject to degradation in the control sample. 2-O Feruloyl-tartaric acid was stable during the whole testing period. All antioxidant treatments greatly improved the stability of caffeic acid derivatives. Stability was dependent upon the concentration of antioxidant added. PMID- 12083868 TI - Ellagic acid and ellagitannins affect on sedimentation in muscadine juice and wine. AB - A mechanism for the formation of water-insoluble sediments in wines and juices made from red and white muscadine grapes (Vitis rotundifolia) was investigated as a function of processing methodology and storage. Sediments are considered quality defects in muscadine grape products, and their presence may influence consumer acceptability and expansion of retail markets. Processing regimes included both hot (70 degrees C) and cold (25 degrees C) press techniques for wine or juice production, and fermentations in contact with grape skins for 3, 5, and 7 days. Relationships between free ellagic acid (FE), total ellagitannins (ET), and total ellagic acid (TE) concentrations were evaluated initially in each product and in sediments that formed during storage for 50 and 120 days at 20 degrees C. Processing techniques influenced initial concentrations of these compounds and the extent of sediment formation. Following storage, juices generally had higher concentrations of FE in sediments compared to wines, but sedimentation was independent of initial FE or TE concentrations. Decreases in ET were observed for hot-pressed juice and skin-fermented wines after storage indicating their hydrolysis during storage and possible contribution to FE in sediments. However, quantitative analysis of the collected sediments revealed that no more than 12% FE by weight was actually present in the sediments, with the remainder consisting of either unidentified compounds or conjugated forms of ellagic acid. This work elucidated a potential mechanism for the presence of FE in muscadine wine and juice sediments through ellagitannin hydrolysis and suggests that sedimentation from mechanisms other than ellagic acid precipitation may also contribute to wine and juice quality. PMID- 12083869 TI - Edible arabinoxylan-based films. 1. Effects of lipid type on water vapor permeability, film structure, and other physical characteristics. AB - Arabinoxylans (AX) are natural fibers extracted from maize bran, an industrial byproduct. To promote this polymer as a food ingredient, development of edible coatings and films had been proposed. Indeed, composite arabinoxylan-based films were prepared by emulsifying a fat: palmitic acid, oleic acid, triolein, or a hydrogenated palm oil (OK35). Lipid effects on water vapor permeability (WVP), surface hydrophobicity (contact angles), lipid particle size, and mechanical properties were investigated. Results showed that OK35-AX emulsion films had the lowest WVP. Emulsified films presented a bimodal particle size distribution; however, the smallest particle mean diameter (0.54 microm) was observed in OK35 AX emulsion films. Contact angles of water comparable to those observed for LDPE films (>90 degrees ) are measured on the OK35-AX film surface. Finally, only triolein-AX emulsion films had elongation higher than films without lipid. These results suggest that OK35 enhances functional properties of AX-based films and should be retained for further research. PMID- 12083870 TI - Suppressive effect of saturated acyl L-ascorbate on the oxidation of linoleic acid encapsulated with maltodextrin or gum arabic by spray-drying. AB - 6-O-Palmitoyl L-ascorbate was added to linoleic acid at various molar ratios of the ascorbate to the acid, the mixtures were emulsified with a maltodextrin or gum arabic solution, and the emulsions were spray-dried to produce microcapsules. At higher molar ratios, the oil droplets in the emulsions were smaller, and the oxidative stabilities of the encapsulated linoleic acid were higher for both the maltodextrin- and gum arabic-based microcapsules. 6-O-Capryloyl, caproyl, and lauroyl L-ascorbates, which were synthesized through lipase-catalyzed condensation in acetone, were also used for the microencapsulation of linoleic acid. Except for capryloyl L-ascorbate, the addition of a saturated acyl ascorbate, especially caproyl ascorbate, to linoleic acid was effective for preparing oil droplets of small particle diameter and for suppressing the oxidation of the encapsulated linoleic acid. PMID- 12083871 TI - Characterization of color fade during frozen storage of red grapefruit juice concentrates. AB - Color changes in red grapefruit juice concentrates during storage at -23 degrees C for 12 months were studied. Concentrate (38 degrees Brix) was packed in both plastic (16 oz) and metal (6 oz) cans. Decrease in red intensity (CIE a) in juice color and slight increases in CIE L*, b*, and hue values from analysis of reconstituted juices were the characteristic color changes in concentrate during frozen storage. With respect to fresh concentrate, juice color in stored concentrate shifted toward the direction between negative DeltaC* and positive DeltaL*, indicating the color became slightly paler. A color difference seems to exist between the two containers, especially for the magnitude of DeltaE*; color changes were more pronounced in concentrates packed in plastic. There are significant changes (P < 0.05) in major carotenoid pigments (beta-carotene and lycopene) in the concentrates. More than 20% loss of lycopene and about 7% loss of beta-carotene occurred with plastic containers after a 12-month period. Regression analysis showed that the rate of decline was about 0.291 ppm per month (r = 0.990) for lycopene compared to 0.045 ppm (r = 0.817) for beta-carotene in concentrate stored in plastic. In the metal can, the same trends were observed but pigment losses were slightly smaller than those with plastic. An estimated shelf life for lycopene was 26.1 months in the metal can compared to 18 months in plastic. Shelf life for beta-carotene was more than 39 months, more than twice that of lycopene in plastic container. PMID- 12083872 TI - Molecular design of antifungal agents. AB - In a rational approach to the design of antifungal agents against Saccharomyces cerevisiae, a series of alkyl gallates (3,4,5-trihydroxybenzoates) were synthesized and assayed. Nonyl gallate (1) was found to be the most effective with a minimum fungicidal concentration (MFC) of 12.5 microg/mL (42 microM), followed by octyl gallate (2) with an MFC of 25 microg/mL (89 microM). These MFCs are little influenced by pH values. A time-kill curve study indicates that nonyl gallate exhibits fungicidal activity against S. cerevisiae at any growing stage. The antifungal activity of nonyl gallate is due primarily to its ability to act as a nonionic surface-active agent (surfactant). The length of the alkyl group is not a major contributor but plays a role in eliciting the activity to a large extent. As far as alkyl gallates are concerned, their antimicrobial spectra and potency depend largely on the hydrophobic portion of the molecules. PMID- 12083873 TI - New monascus metabolite isolated from red yeast rice (angkak, red koji). AB - Red yeast rice (angkak, red koji) obtained as cultures of Monascus purpureus on rice was extracted and analyzed by HPLC. In addition to the known red, orange, and yellow pigments and the mycotoxin citrinin, a new Monascus metabolite was detected. It is present in the original red yeast rice and formed in higher amounts when red yeast rice is heated. High-resolution mass spectrometry indicated the molecular formula C(15)H(12)O(4). The chemical structure was elucidated by analysis of NMR data. The new compound, named monascodilone, is characterized by a propenyl group on a pyrone ring, an aromatic ring, and a gamma lactone group. PMID- 12083874 TI - Influence of herbicide structure, clay acidity, and humic acid coating on acetanilide herbicide adsorption on homoionic clays. AB - Adsorption of chloroacetanilide herbicides on homoionic montmorillonite was studied by coupling batch equilibration and FT-IR analysis. Adsorption decreased in the order metolachlor > acetochlor > alachlor > propachlor on Ca(2+)- or Mg(2+)-saturated clays and in the order metolachlor > alachlor > acetachlor > propachlor on Al(3+)- or Fe(3+)-saturated clays. FT-IR spectra showed that the carbonyl group of the herbicide molecule was involved in bonding. For the same herbicide, adsorption of alachlor, acetachlor, and metolachlor on clay followed the order Ca(2+) approximately Mg(2+) < Al(3+) < or = Fe(3+), which coincided with the increasing acidity of homoionic clays. Adsorption of propachlor, however, showed an opposite dependence, suggesting a different governing interaction. In clay and humic acid mixtures, herbicide adsorption was less than that expected from independent additive adsorption by the individual constituents, and the deviation was dependent on the clay-to-humic acid ratio, with the greatest deviation consistently occurring at a 60:40 clay-to-humic acid ratio. PMID- 12083875 TI - Volatilization of trifluralin, atrazine, metolachlor, chlorpyrifos, alpha endosulfan, and beta-endosulfan from freshly tilled soil. AB - The volatile and soil loss profiles of six agricultural pesticides were measured for 20 days following treatment to freshly tilled soil at the Beltsville Agricultural Research Center. The volatile fluxes were determined using the Theoretical Profile Shape (TPS) method. Polyurethane foam plugs were used to collect the gas-phase levels of the pesticides at the TPS-defined critical height above a treated field. Surface-soil (0-8 cm) samples were collected on each day of air sampling. The order of the volatile flux losses was trifluralin > alpha endosulfan > chlorpyrifos > metolachlor > atrazine > beta-endosulfan. The magnitude of the losses ranged from 14.1% of nominal applied amounts of trifluralin to 2.5% of beta-endosulfan. The daily loss profiles were typical of those observed by others for volatile flux of pesticides from moist soil. Even though heavy rains occurred from the first to third day after treatment, the majority of the losses took place within 4 days of treatment, that is, 59% of the total applied atrazine and metolachlor and >78% of the other pesticides. Soil losses generally followed pseudo-first-order kinetics; however, leaching due to heavy rainfall caused significant errors in these results. The portion of soil losses that were accounted for by the volatile fluxes was ordered as follows: alpha-endosulfan, 34.5%; trifluralin, 26.5%; chlorpyrifos, 23.3%; beta endosulfan, 14.5%; metolachlor, 12.4%; and atrazine, 7.5%. PMID- 12083876 TI - Production, characterization, and effects on tomato of humic acid-like polymerin metal derivatives from olive oil mill waste waters. AB - The dark polymeric organic fraction rich in potassium recovered from olive oil mill waste waters (OMWW) and named polymerin and the potassium salified deglycosylated polymerin derivative (K-SDpolymerin) were easily transformed into their metal derivatives by saturation with various metals, including Na, Cu, Zn, Mn, Fe, and Al. Saturated metal polymerins were characterized by diffuse reflectance infrared Fourier transform spectroscopy and atomic absorption spectrometry. Tests on tomato plants of the various polymerins showed that only the soluble polymerin, K-SDpolymerin, and the insoluble Mn-SDpolymerin were significantly toxic. The toxic effects of OMWW on tomato at the original concentration and diluted 1:10 were much stronger than those of any polymerin. The possible exploitation of polymerins as bioamendments and/or metal biointegrators as a functon of their phytotoxic effects, their humic acid-like nature, and their richness in macro- and micronutrient metals is also discussed. PMID- 12083877 TI - Aroma biosynthesis in strawberry: s-adenosylmethionine:furaneol o methyltransferase activity in ripening fruits. AB - Among the most important volatile compounds in the aroma of strawberries are 2,5 dimethyl-4-hydroxy-3(2H)-furanone (Furaneol) and its methoxy derivative (methoxyfuraneol, mesifuran). Three strawberry varieties, Malach, Tamar, and Yael, were assessed for total volatiles, Furaneol, and methoxyfuraneol. The content of these compounds sharply increased during fruit ripening, with maximum values at the ripe stage. An enzymatic activity that transfers a methyl group from S-adenosylmethionine (SAM) to Furaneol sharply increases during ripening of strawberry fruits. The in vitro generated methoxyfuraneol was identified by radio TLC and GC-MS. The partially purified enzyme had a native molecular mass of approximately 80 kDa, with optimum activity at pH 8.5 and 37 degrees C. A high apparent K(m) of 5 mM was calculated for Furaneol, whereas this enzyme preparation apparently accepted as substrates other o-dihydroxyphenol derivatives (such as catechol, caffeic acid, and protocatechuic aldehyde) with much higher affinities (K(m) approximately 105, 130, and 20 microM, respectively). A K(m) for SAM was found to be approximately 5 microM, regardless of the acceptor used. Substrates that contained a phenolic group with only one OH group, such as p coumaric and trans-ferulic acid, as well as trans-anol and coniferyl alcohol, were apparently not accepted by this activity. It is suggested that Furaneol methylation is mediated by an O-methyltransferase activity and that this activity increases during fruit ripening. PMID- 12083878 TI - Catalytic properties of alcohol acyltransferase in different strawberry species and cultivars. AB - The substrate specificity of alcohol acyltransferase (AAT) enzymes from different strawberry varieties was studied. Proteins with AAT activity from fruits of Fragaria x ananassa Duch. cv. Oso Grande were purified to apparent homogeneity and used for kinetic studies with different straight-chain alcohols and acyl CoAs. K(m) values obtained for Oso Grande enzyme with six different alcohols, using acetyl-CoA as cosubstrate, decreased with increasing length of the alcohol chain. In similar experiments the increase in the acyl-CoA carbon chain was also found to be correlated with a higher substrate specificity. Heptanol (K(m) = 0.73 mM) and hexanoyl-CoA (K(m) = 0.41 mM) were the best substrates for Oso Grande AAT. Comparative catalytic studies were carried out with AAT partially purified extracts from the wild type Fragaria vesca and five commercial strawberry varieties: Tudnew, Carisma, Camarosa, Sweet Charlie, and Eris. The specificities of these enzymes toward five selected alcohols and acyl-CoAs reflected interesting cultivar differences. PMID- 12083879 TI - Biosynthesis of strawberry aroma compounds through amino acid metabolism. AB - The fate of amino acids in relation to aroma biogenesis was studied in strawberries using the in vitro growth approach. This fruit presented differences in the level of metabolization for different amino acids. Incubations of strawberries with L-isoleucine gave rise to an increase of fourteen compounds in this fruit aroma, either not detected previously or constituents of strawberry aroma. However, L-valine incubations did not provide a significant change in this fruit aroma. Strawberry feeding with L-isoleucine resulted in a 7-fold increase in the sum of 2-methylbutanoate esters, and a double production of 2-methylbutyl esters compared to those of control fruits. Around 94% of the ester increase corresponded to 2-methylbutanoates, with ethyl 2-methylbutanoate being the most representative compound (92%). On the other hand, among the 2-methylbutyl esters, comprising around 6% of total aroma volatiles increase, 2-methylbutyl acetate was the major compound (95%) arising from L-isoleucine strawberry feeding. The role of enzymatic activities within the amino acid metabolic pathway in strawberry fruits is discussed. PMID- 12083880 TI - Effect of storage on some volatile aroma compounds in fresh-cut cantaloupe melon. AB - Changes in volatile aroma constituents of fresh-cut cantaloupe melon with storage were determined by headspace solid-phase microextraction gas chromatography-mass spectrometry. The compounds isolated from the fruit immediately after cutting were predominantly aliphatic and aromatic esters. Storage of fruit at 4 degrees C caused a considerable decrease in concentration of esters and synthesis of the terpenoid compounds beta-ionone and geranylacetone over a period of 24 h. This change in the volatile profile with storage is consistent with that of a stress induced defense response in the cut fruit as an adaptation process to tissue exposure and cell disruption. The same effect occurred in fruit stored at 22 degrees C and in those treated with sodium azide and ascorbic acid prior to storage. Fruit treated with ascorbic acid and sodium azide had higher concentrations of beta-ionone and geranylacetone and retained these compounds better with storage time. The reduction of esters appears to be an important early reaction step in the loss of freshness during storage of fresh-cut cantaloupe. PMID- 12083881 TI - Chemical characterization of the aroma of Grenache rose wines: aroma extract dilution analysis, quantitative determination, and sensory reconstitution studies. AB - The aroma of a Grenache rose wine from Calatayud (Zaragoza, Spain) has been elucidated following a strategy consisting of an aroma extract dilution analysis (AEDA), followed by the quantitative analysis of the main odorants and the determination of odor activities values (OAVs) and, finally, by a series of reconstitution and omission tests with synthetic aroma models. Thirty-eight aroma compounds were found in the AEDA study, 35 of which were identified. Twenty-one compounds were at concentrations higher than their corresponding odor thresholds. An aroma model prepared by mixing the 24 compounds with OAV > 0.5 in a synthetic wine showed a high qualitative similarity with the aroma of the rose wine. The addition of compounds with OAV < 0.5 did not improve the model, whereas the aroma of a model containing only odorants with OAV > 10 was very different from that of the wine. Omission tests revealed that the most important odorant of this Grenache rose wine was 3-mercapto-1-hexanol, with a deep impact on the wine fruity and citric notes. The synergic action of Furaneol and homofuraneol also had an important impact on wine aroma, particularly in its fruity and caramel notes. The omission of beta-damascenone, which had the second highest OAV, caused only a slight decrease on the intensity of the aroma model. Still weaker was the sensory effect caused by the omission of 10 other compounds, such as fatty acids and their ethyl esters, isoamyl acetate, and higher alcohols. PMID- 12083882 TI - Characterization of 2-methyl-4-amino-5-(2-methyl-3-furylthiomethyl)pyrimidine from thermal degradation of thiamin. AB - Thiamin hydrochloride was thermally degraded in phosphate buffer (pH 6.5) at 110 degrees C for 2 h. A major decomposition product was isolated by column chromatography and structurally identified by spectrometric techniques ((1)H NMR, (13)C NMR, 2D NMR, and MS) as 2-methyl-4-amino-5-(2-methyl-3 furylthiomethyl)pyrimidine (MAMP). The possible formation pathway of MAMP was studied using two model systems. It is proposed that MAMP is formed by nucleophilic attack of 2-methyl-3-furanthiol on the thiamin. PMID- 12083883 TI - Phytochemical induction by herbivores could affect quality of essential oils from aromatic plants. AB - Plant tissues may show chemical changes following herbivory. In aromatic plants such changes could affect the specific compounds on which commercial exploitation is based. This possibility was analyzed for Mintosthachys mollis, a member of the Lamiaceae native to Central Argentina with medicinal and aromatic uses in the region, and two types of insect herbivores: a leaf miner and a gall insect. Analysis of the essential oils of mined/undamaged leaves, as well as leaves from stems with and without galls, revealed changes in concentrations of the two main monoterpenes. A decrease in pulegone concentration was associated with both types of insect damage, whereas menthone increased significantly only in mined leaves. Inducible chemical changes in aromatic and medicinal plants may be common and widespread; their economic implications deserve investigation. PMID- 12083884 TI - Thermal degradation studies of glucose/glycine melanoidins. AB - Nondialyzable and water-insoluble melanoidins, isolated from a glucose/glycine model reaction mixture, which was prepared in a standardized way according to the guidelines of the COST Action 919, were heated at different temperatures ranging from 100 to 300 degrees C. Among the volatile compounds, which were analyzed by SPME and GC-MS, pyrazines, pyridines, pyrroles, and furans were detected. In general, total amounts of volatile compounds increased with the temperature. When water-insoluble melanoidins were heated, especially at higher temperatures, this resulted in a higher diversity of isolated compounds. For furans, pyrroles, pyrazines, and carbonyl compounds a maximum was observed in the case of high molecular weight melanoidins around 200-220 degrees C. Pyridines and total oxazoles, however, were generated in higher yields with increasing temperatures. These results demonstrate the possibility of producing some flavor-significant volatiles from heated standard melanoidins at temperatures relevant to food preparation and contribute to the flavor aspects originating from melanoidins. PMID- 12083885 TI - Aroma quality differentiation of pyrazine derivatives using self-organizing molecular field analysis and artificial neural network. AB - The encoding of various aroma impressions and the distinction between different aroma qualities are unsolved problems, as differences between aroma impressions can be described only in a qualitative but not in a quantitative manner. As a consequence, classifications of various aroma qualities cannot easily be performed by standard QSAR methods. To find a proper way to encode aroma impressions for SAR studies, a total of 50 pyrazine-based aroma compounds showing the aroma quality of earthy, green-earthy, or green are analyzed. Special attention is thereby turned on the mixed aroma impression green-earthy. Classifications on the whole data set as well as on smaller subsets are calculated using self-organizing molecular field analysis (SOMFA) and artificial neural networks (ANNs). SOMFA classifies between two or three aroma impressions, leading to models satisfying in predictive power. ANN analysis using multilayer perceptron network architecture with one hidden layer and nominal output as well as genetic regression neural network) with two hidden layers and numerical output both lead to a rather good performance rate of 94%. PMID- 12083886 TI - Sulfur aroma precursor present in S-glutathione conjugate form: identification of S-3-(hexan-1-ol)-glutathione in must from Vitis vinifera L. cv. Sauvignon blanc. AB - When Sauvignon blanc or Gros Manseng grape must was percolated through an immobilized gamma-glutamyltranspeptidase column, there was a significant increase in the concentration of S-3-(hexan-1-ol)-L-cysteine, the precursor of 3 mercaptohexan-1-ol, a compound that contributes to the varietal aroma of wines made from these grapes. Low- and high-resolution liquid secondary ion mass sepectrometry (LSIMS) analyses established the presence of S-3-(hexan-1-ol) glutathione in Sauvignon blanc must. The identification of this compound suggests that the S-3-(hexan-1-ol)-L-cysteine in grapes is produced by the catabolism of S 3-(hexan-1-ol)-glutathione. As is the case in other plant or animal organisms, S glutathione conjugates may be involved in certain detoxification systems in vines. PMID- 12083887 TI - Formation of sulfur aroma compounds in reaction mixtures containing cysteine and three different forms of ribose. AB - The headspace volatiles produced from buffered and unbuffered cysteine model systems, containing inosine 5'-monophosphate, ribose 5-phosphate, or ribose, were examined by GC-MS. Sulfur compounds dominated the volatiles of all systems and included mercaptoketones, furanthiols, and disulfides. The inosine monophosphate systems produced much lower quantities of volatiles than ribose phosphate or ribose systems. In the systems buffered with phosphate or phthalate buffers, both ribose and ribose phosphate systems gave similar quantities of sulfur volatiles. However, in the absence of buffer, the ribose system was relatively unreactive, especially for volatiles formed via the 2,3-enolization route in the Maillard reaction, where 4-hydroxy-5-methyl-3(2H)-furanone is a key intermediate. A number of keto-enol tautomerisms, which are known to be acid-base-catalyzed, occur in the 2,3-enolization route. This may explain the catalysis of the ribose systems by the buffers. In the ribose phosphate systems, however, Maillard mechanisms probably played a less important role, because ribose 5-phosphate readily dephosphorylated to give 4-hydroxy-5-methyl-3(2H)-furanone on heating and thus provided an easier route to aroma compounds than the Maillard reaction. PMID- 12083888 TI - Lipase-assisted generation of 2-methyl-3-furanthiol and 2-furfurylthiol from thioacetates. AB - Enzymatic hydrolysis of S-3-(2-methylfuryl) thioacetate and S-2-furfuryl thioacetate using lipase from Candida rugosa produced 2-methyl-3-furanthiol and 2 furfurylthiol, respectively. When reactions were carried out at room temperature and pH 5.8, 2-methyl-3-furanthiol was produced in a optimal yield of 88% after 15 min of reaction, whereas 2-furfurylthiol was obtained in a yield of 80% after 1 h of reaction time. Enzymatic hydrolysis was also performed in n-hexane, n-pentane, and water/propylene glycol mixture. The reaction rates in these media were slower as compared to those in aqueous medium; however, the reaction yields were quite similar. As expected, the stability of the generated 2-methyl-3-furanthiol and 2 furfurylthiol was better in n-hexane, n-pentane, and the water/propylene glycol mixture as compared to that in water or phosphate buffer. PMID- 12083889 TI - Clarification of juice by thermolabile valencia pectinmethylesterase is accelerated by cations. AB - Pectinmethylesterase (PME) was isolated from Valencia orange pulp and added to reconstituted juice at 1.2 units/mL of juice in the presence or absence of cations at 4.2 or 16.7 mM. The percent transmittance (%T) of control juices with no added PME or cation did not clarify. The %T of juices with added PME and added cation was 45-55% by the second day. Increases in the average particle size was observed with PME- or cation-added juices and preceded increases in %T. Most likely, cations displaced PME from an inactive pectin substrate complex and increased clarification. PME, in the absence of cations, increased particle size but did not affect %T, suggesting a direct interaction of PME with cloud particles. PMID- 12083890 TI - Variations in the profile and content of anthocyanins in wines made from cabernet sauvignon and hybrid grapes. AB - To detect adulteration of wine, it has been proposed that the ratio of acetylated to p-coumaroylated conjugates of nine characteristic anthocyanins can be used to determine whether a wine is derived from Cabernet Sauvignon or hybrid grapes. If the ratio is >3, then a wine is classified as being derived from Cabernet Sauvignon grapes. This test has significant commercial implications as it is being used to decide whether Cabernet Sauvignon-labeled wines are genuine and can be imported into Germany. To assess whether this is a valid approach, 24 wines were analyzed, 4 of which were made from hybrids and 20 from Cabernet Sauvignon, with vintages ranging from 1993 to 2000. Only 13 of the Cabernet Sauvignon wines contained all nine of the "characteristic" anthocyanins, and the ratio of acetylated to p-coumaroylated derivatives varied from 1.2 to 6.5. It is evident that the use of the anthocyanin ratio method is flawed and that examination of the whole anthocyanin profile and/or investigation of the proportion of monoglucoside and acetylated anthocyanins is a better approach to distinguish between hybrid and Cabernet Sauvignon wines. PMID- 12083892 TI - Kinetics of mushroom tyrosinase inhibition by quercetin. AB - The effects of quercetin on the activity of mushroom tyrosinase were studied. The equilibrium constants for this inhibitor binding with the enzyme molecule were established. The inhibition mechanism obtained from Lineweaver-Burk plots show that quercetin is a competitive inhibitor. In the time course of the oxidation of L-3,4-dihydroxyphenylalanine (L-DOPA) catalyzed by the enzyme in the presence of different concentrations of quercetin, the rate decreased with increasing time until a straight line was approached. The inhibition of tyrosinase by quercetin is a slow and reversible reaction with residual enzyme activity. The microscopic rate constants were determined for the reaction of quercetin with the enzyme. PMID- 12083891 TI - Partial purification and characterization of pectin methylesterase from acerola (Malpighia glabra L.). AB - The enzyme pectin methylesterase (PME) is present in acerola fruit and was partially purified by gel filtration on Sephadex G-100. The results of gel filtration showed different PME isoforms. The total PME (precipitated by 70% salt saturation) and one of these isoforms (fraction from Sephadex G-100 elution) that showed a molecular mass of 15.5 +/- 1.0 kDa were studied. The optimum pH values of both forms were 9.0. The total and the partially purified PME showed that PME specific activity increases with temperature. The total acerola PME retained 13.5% of its specific activity after 90 min of incubation at 98 degrees C. The partially purified acerola (PME isoform) showed 125.5% of its specific activity after 90 min of incubation at 98 degrees C. The K(m) values of the total PME and the partially purified PME isoform were 0.081 and 0.12 mg/mL, respectively. The V(max) values of the total PME and the partially purified PME were 2.92 and 6.21 micromol/min/mL/mg of protein, respectively. PMID- 12083893 TI - Improvement of gel properties of dried egg white by modification with galactomannan through the Maillard reaction. AB - The effects of Maillard reaction on gel properties of dried egg white (DEW) with galactomannan (GM) were investigated. Maillard-reacted DEW (MDEW) was prepared by dry-heating a mixture with a weight ratio of 1:4 of GM to DEW at 60 degrees C and 65% relative humidity. The modification of amino groups and polymerization of DEW proteins dry-heated with GM proceeded with increasing the dry-heating time. The covalent attachment of GM to DEW was confirmed from SDS-PAGE analysis. Gel strength and water-holding capacity of MDEW gels were higher than those of DEW dry-heated without GM (control DEW) and reached maximum after 3 days of dry heating. The appearance of MDEW gels became transparent with increasing the dry heating time, but control DEW gels were still turbid. MDEW dry-heated for 3 days was almost soluble even after heating of its solution at 90 degrees C, whereas control DEW proteins precipitated. The modification of DEW with GM through the Maillard reaction was an effective method to make a firm and transparent gel from DEW at broader range of pH and NaCl concentration of the medium. PMID- 12083894 TI - Kinetic parameters for the thermal inactivation of quality-related enzymes in carrots and potatoes. AB - Kinetic parameters for the thermal inactivation of several enzymes in carrot and potato homogenates have been determined. In carrots the most heat-resistant activity was polygalacturonase, followed by peroxidase and pectinmethylesterase. In potatoes peroxidase was the most resistant, followed by pectin methylesterase, polyphenol oxidase, and lipoxygenase. There were several notable similarities between the inactivation kinetics in the two vegetables. In both cases peroxidase activity gave simple first-order inactivation kinetics but yielded a curved Arrhenius plot for the temperature dependence. Pectin methylesterase in both commodities consisted of a labile and a resistant form. The relative amounts of the two forms and the temperature dependences for their inactivation were also similar. PMID- 12083895 TI - Improvement of flour quality through carbohydrases treatment during wheat tempering. AB - Wheat flour is obtained by the milling process, which includes several steps such as cleaning, tempering, and milling. In the tempering the moisture content of wheat grains is increased to 15.5% by adding an adequate amount of water. The addition of different enzymes (cellulase, xylanase, and beta-glucanase) to the tempering solution has been tested in order to modify the quality of the resulting flour. Rheological and fermentative properties were measured by the farinograph, amylograph, and rheofermentometer. The data show that the technological parameters of the resulting flours were greatly modified by the addition of enzymes to the tempering solution. The quality of the fresh bread obtained from the carbohydrase-treated wheat was improved with regard to specific bread volume, bread shape, and crumb firmness. This method is revealed as an excellent tool to ensure a good distribution of the enzymes in the resulting flour, to control dosage during milling, and to obtain flour of specific characteristics according to their final use. PMID- 12083896 TI - Comparative efficiencies of isopropyl and tert-butyl alcohols for extracting zeins from maize endosperm. AB - Protein of endosperm of maize grains originating from three wild-type inbreds and their opaque-2 versions were solubilized in diverse extracts (E) by the sequential use of 0.5 M NaCl, water (E(1,2)), alcohol plus a reducing agent (E(3)), and salt plus a reducing agent (E(4)). Zeins were isolated in extracts E(3) and E(4) obtained by using 55% (w/w) isopropyl alcohol (i-PrOH) + 0.2% dithiothreitol (DTT) followed by 0.5 M NaCl + 0.2% DTT buffered at pH 10 or 60% tert-butyl alcohol (t-BuOH) + 0.2% DTT followed by 0.5% sodium acetate + 0.2% DTT in 30% t-BuOH. For a given genotype the percentage of extracted zeins was independent of the nature of the alcohol. The latter had a slight effect on the respective magnitude of E(3) and E(4): E(3) increased at the expense of E(4) when t-BuOH was substituted to i-PrOH for their isolation. The percentage of the total endosperm nitrogen present in E(3) + E(4) was identical to that of fractions F(II) + F(III) + F(IV) isolated according to the classical Landry-Moureaux extraction procedure. SDS-PAGE analysis revealed the presence of all types of zeins (alpha, beta, gamma, and delta) in E(3) and F(III), residual zeins in E(4) isolated with t-BuOH, and streaking only in E(4) and F(IV) isolated with NaCl at pH 10. The data together with those of the literature were discussed with regard to the influence of procedure on the yield of zeins using alcoholic extraction. PMID- 12083897 TI - Conjugated linoleic acid isomers differ in their free radical scavenging properties. AB - Conjugated linoleic acid (CLA) isomers were investigated for free radical scavenging properties against the stable 2,2-diphenyl-1-picryhydrazyl radical (DPPH(*)) by electron spin resonance (ESR) spectrometry and spectrophotometric methods. ESR measurements confirmed that both c9,t11-CLA and t10,c12-CLA directly reacted with and quenched DPPH radicals, whereas spectrophotometric analysis demonstrated that c9,t11-CLA and t10,c12-CLA differed in their kinetic and thermodynamic properties in reacting with DPPH radicals. t10,c12-CLA was shown to exhibit a greater initial velocity in CLA-DPPH radical reactions at levels of 2.5 80 mg/mL, and c9,t11-CLA scavenged more DPPH radicals at steady state. Similar dose and time relationships were observed for both isomers. In addition, a mixture of c9,t11- and t10,c12-CLA isomers demonstrated a greater initial velocity in quenching DPPH radicals than either isomer alone on the same concentration basis, suggesting that a synergistic effect between CLA isomers existed in their reactions with DPPH radicals. These results support the conclusion that individual CLA isomers differ in their biological actions and indicate that interaction(s) between isomers may contribute to their beneficial effects. PMID- 12083899 TI - Nutritional value of a highly digestible sorghum cultivar for meat-type chickens. AB - The nutritional value of a newly discovered sorghum mutant cultivar (P851171), with high in vitro protein digestibility, was compared to those of corn and two normal sorghums (P721N and 611Y) in two chick feeding trials. Although 8-20 day protein efficiency ratios and net protein ratios of all three sorghums were inferior to those of corn, P851171 and 611Y had markedly greater mean true amino acid digestibilities (TAAD) than either corn or P721N. In a subsequent 42-day experiment, all three sorghums supported weight gains equal to those of the corn fed chicks. Feeding suboptimal levels of dietary protein resulted in reduced weight gains and no observed benefits of P851171 or 611Y. Furthermore, chicks fed P851171 exhibited poorer feed/gain values as compared to those fed the other cereals. It is possible that the starch content/carbohydrate profile of P851171 was inferior to that of the other sorghums, which offset its superior TAAD and resulted in poorer broiler performance. PMID- 12083898 TI - Furosine as indicator of maillard reaction in jams and fruit-based infant foods. AB - To investigate the presence of furosine in commercial samples of jams and fruit based infant foods a simple method by ion-pair reversed-phase liquid chromatography is described. The yield of furosine during the hydrolysis with hydrochloric acid was optimized. The reproducibility in the repeatability and recovery of the method, expressed in relative standard deviation percentages, proved to be in the ranges of 4.1-8.3% and 1.0-4.4%, respectively. The recovery percentages of furosine varied between 86.7 and 95.3%. The obtained results support the suitability of the method. Furosine was detected in all studied samples. Although a high variability in the content of furosine was noticed, in general terms, the lowest levels of furosine were observed in samples of fruit based infant foods and the highest were observed in jams of more than 60% sugar. These results could be due to different heat treatment, storage conditions, and/or differences in the values of water activity (a(w)) and amounts of sugar. The results obtained in the present paper point out the usefulness of furosine as an indicator of Maillard reaction for jams and fruit-based infant foods. PMID- 12083900 TI - Molecular "chaperones" guide the spontaneous formation of a 15-component hydrogen bonded assembly. AB - Chaperones are small molecules that assist in the folding of naturally occurring peptides. There are no examples of small molecules acting as chaperones in the self-assembly of synthetic noncovalent assemblies. In this communication we describe an unprecedented example of the "chaperone effect" in the noncovalent synthesis of organic nanostructures. Tetrarosette assemblies 2(3).(BuCYA)(12) form quantitatively in CHCl(3) at room temperature upon mixing tetramelamine 2 with N-butylcyanurate (BuCYA) in the presence of 5,5-diethylbarbituric acid (DEB). Without the DEB units present, only oligomeric assemblies are formed that cannot rearrange to the tetrarosettes by themselves. The DEB units act as molecular "chaperones" by preorganizing the tetramelamine units for the spontaneous assembly of the tetrarosette structure. PMID- 12083902 TI - Fabrication of hollow palladium spheres and their successful application to the recyclable heterogeneous catalyst for suzuki coupling reactions. AB - Novel palladium hollow spheres were synthesized using silica spheres as a template, and they were successfully applied as recyclable heterogeneous catalysts for Suzuki cross coupling reactions. PMID- 12083901 TI - Asymmetric induction in the Neber rearrangement of simple ketoxime sulfonates under phase-transfer conditions: experimental evidence for the participation of an anionic pathway. AB - Phase-transfer catalysis has been successfully utilized for the Neber rearrangement of simple ketoxime sulfonates. For instance, treatment of (Z)-1a with p-toluenesulfonyl chloride (1.2 equiv), tetrabutylammonium bromide (5 mol %) and MeOH (10 equiv) in toluene-50% KOH aqueous solution (volume ratio = 3:1) at 0 degrees C for 2 h, and subsequent benzoylation followed by acidic hydrolysis afforded the protected alpha-amino ketone 2a in 80% isolated yield. On the basis of this finding, asymmetric Neber rearrangement with ketoxime (Z)-1a was examined under phase-transfer conditions using structurally rigid, C(2)-symmetric chiral quaternary ammonium bromide 3 as catalyst. The desired 2a was obtained in 80% yield and 51% ee. This represents the first experimental demonstration of the substantial involvement of anion pathway in the Neber rearrangement of simple ketoxime sulfonates. Importantly, the reaction with (E)-1a under otherwise similar conditions afforded racemic 2a in 61% yield. Moreover, enhanced enantioselectivity (63% ee) was observed in the rearrangement of the oxime sulfonate derived from (Z)-1b, and notably, use of mesitylene in place of toluene further increased the enantioselectivity to 70% ee. Our approach provides not only a new mechanistic insight but also an opportunity for extending the full synthetic utility of this classical yet useful rearrangement. PMID- 12083903 TI - Enzymatic manipulation of DNA-nanomaterial constructs. AB - The demonstration and control of biofunction between inorganic nanomaterials and biological scaffolding is crucial to the development of the field of biomaterials. Although unique hierarchical structures can be generated, the impact of nanosized materials on the biological activity of DNA-protein interactions is relatively unknown. Using highly selective proteins that induce sequence-specific conformational perturbations within DNA, we demonstrate the absolute maintenance of biofunction for biomaterials composed of duplex DNA appended with 1.4-nm Au particles. Enzyme activity and DNA binding affinities (K(d)) are unaltered by the nanoparticle-DNA conjugates. Our results provide a foundation for interfacing more complex and diverse protein-DNA-systems. PMID- 12083904 TI - Hydrogen bonding to Trp beta37 is the first step in a compound pathway for hemoglobin allostery. AB - Human hemoglobin is widely thought to change from the R to the T quaternary structure in a single rate process requiring tens of microseconds. Here we present kinetic evidence that the R --> T allosteric pathway in hemoglobin requires more than one step. We use magnetic circular dichroism (MCD) spectroscopy of the aromatic amino acid bands to show that formation of a tryptophan-aspartate hydrogen bond in the hinge region of the dimer-dimer interface is part of an obligatory R --> T step proceeding more than a factor of 10 faster than the kinetic step previously identified in heme-band absorption studies. PMID- 12083905 TI - Synthesis and crystal structure of ionic multicomponent complex: ([Cr(I)(PhH)(2)].+))(2)[Co(II)TPP(C(60)(CN)(2))]-[C(60)(CN)(2)](.-).3(o C(6)H(4)Cl(2)) containing C(60)(CN)(2).- radical anion and sigma-bonded diamagnetic Co(II)TPP(C(60)(CN)(2)) -anion. AB - The ionic multicomponent complex complex: ([Cr(I)(PhH)(2)].+))(2)[Co(II)TPP(C(60)(CN)(2))]-[C(60)(CN)(2)](.-).3(o C(6)H(4)Cl(2)) (Co(II)TPP: cobalt (II) tetraphenylporphyrin; Cr(PhH)(2): bis(benzene)chromium; o-C(6)H(4)Cl(2): o-dichlorobenzene) containing CoTPP(C(60)(CN)(2)- anion and C(60)(CN)(2).- radical anion was obtained. The complex has the cage structure with channels, which accommodate Cr(I)(PhH)(2)(.+) and o-C(6)H(4)Cl(2) molecules. For the first time the sigma-bonding of Co(II)TPP to the fullerene radical anion with the essentially shortened Co.C(C(60)(CN)(2)) contact of 2.282 A is observed. The sigma-bonding results in the diamagnetism of Co(II)TPP(C(60)(CN)(2))(-) anion. The nonbonded C(60)(CN)(2)(.-) radical anion retains both the C(2)(v)symmetry and the shape of the molecule. The length of the C(triple bond)N bonds is 1.141 and 1.152 A. PMID- 12083907 TI - Kinetic and thermodynamic preferences for the diastereoselective oxidative addition of H(2) to trans-Ir(PR(3))(2)(CO)Cl: monodentate chiral phosphines may impart exceptional degrees of diastereoselectivity. AB - Studies on square planar iridium complexes of the type trans-Ir(PR(3))(2)(CO)Cl, where PR(3) is PhP[(C(5)Me(4))](2), PhP[Me(2)C(4)H(6)], or PhP[Pr(i)(2)C(4)H(6)], demonstrate that monodentate chiral phosphines impart exceptional degrees of diastereoselectivity in the oxidative addition of H(2). Thus, the oxidative addition of H(2) to the two faces of the meso isomer (R,S)-trans Ir(PR(3))(2)(CO)Cl proceeds with a kinetic diastereoselectivity which exceeds that for related square planar iridium complexes employing bidentate chiral phosphine ligands. Furthermore, the kinetically favored dihydride is not favored thermodynamically, and the magnitude of the inversion of the kinetic and thermodynamic selectivities is greater than has previously been observed using bidentate phosphines. PMID- 12083906 TI - A versatile prodrug approach for liposomal core-loading of water-insoluble camptothecin anticancer drugs. AB - We describe a versatile prodrug strategy for loading the liposomal lumen with water-insoluble camptothecins. The procedure involves conversion of an active camptothecin analogue to a 20-OR omega-aminoalkanoanic ester prodrug in which R = CO[CH(2)](n)()NH(2) and n = 1-3. The basic amino group of the prodrug serves three roles. First, at pH ranges of 3-5, the amine enhances aqueous solubility. Second, it enhances responsiveness to a transmembrane ammonium sulfate gradient across the liposomal bilayer, thereby facilitating active loading of the agent into the liposomal aqueous core. Third, at a physiological pH of 7 or above (the pH to be encountered following drug release at the tumor site), the nucleophilicity of the amine manifests itself and cyclization to the C-21 carbonyl carbon occurs. This cyclization triggers a rapid and convenient nonenzymatic decomposition process that releases active camptothecin. Accordingly, this novel liposomal approach offers a potential system for tumor targeting prodrugs of many water-insoluble camptothecins, including the highly lipophilic and clinically attractive analogues SN-38, 9-nitrocamptothecin and DB 67. The rate of formation of the active agent at the tumor site can be controlled through the selection of n (the length of the alkyl spacer group). PMID- 12083908 TI - Interfacial chemistries for nanoscale transfer printing. AB - We describe a patterning technique that uses self-assembled monolayers and other surface chemistries for guiding the transfer of material from relief features on a stamp to a substrate. This purely additive contact printing technique is capable of nanometer resolution. Pattern transfer is fast and it occurs at ambient conditions. We illustrate the versatility of this method by printing single-layer metal patterns with feature sizes from a few tens of microns to a few tens of nanometers. We also demonstrate its use for patterning, in a single step, metal/dielectric/metal multilayers for functional thin film capacitors on plastic substrates. PMID- 12083909 TI - A cyano-bridged single-molecule magnet: slow magnetic relaxation in a trigonal prismatic MnMo(6)(CN)(18) cluster. AB - We report the synthesis of the first well-documented example of a cyano-bridged single-molecule magnet. An assembly reaction parallel to that employed in producing the trigonal prismatic [(Me(3)tacn)(6)MnCr(6)(CN)(18)](2+) (Me(3)tacn = N,N',N"-trimethyl-1,4,7-triazacyclononane) cluster affords K[(Me(3)tacn)(6)MnMo(6)(CN)(18)](ClO(4))(3) (1), containing an analogous molybdenum(III)-substituted cluster. Fits to the DC magnetic susceptibility and magnetization data for 1 show that the MnMo(6) cluster possesses weak antiferromagnetic coupling (J = -6.7 cm(-1)), leading to an S = (13)/(2) ground state with significantly enhanced magnetic anisotropy (D = -0.33 cm(-1) and E = 0.018 cm(-1)). Consistent with these results, AC magnetic susceptibility measurements show the molecule to exhibit slow magnetic relaxation indicative of a single-molecule magnet with an energy barrier of 10 cm(-1) for spin reversal. PMID- 12083910 TI - Enantioselective guest exchange in a chiral resorcin[4]arene cavity. AB - Gas-phase proton-bound complexes between a chiral resorcin[4]arene and some representative amino acids, that is, L- and D-alanine or L- and D-serine, were generated in the source of a Fourier transform ion cyclotron resonance mass spectrometer. Gas-phase exchange of the amino acid from the diastereomeric complexes with the enantiomers of 2-butylamine exhibits a significant enantioselectivity, which depends not only upon the configuration of the leaving guest but also on that of the incoming amine. These findings, coupled with molecular dynamic calculations, point to the observed gas-phase enantioselectivity as determined by the effects of the resorcin[4]arene chiral cavity upon the diastereomeric exchange transition structures. PMID- 12083912 TI - Et(2)Al(+) alumenium ion-like chemistry. synthesis and reactivity toward alkenes and alkene oxides. AB - Inert weakly coordinating carborane anions, CB(11)H(6)X(6)(-) (X = Cl, Br), allow access to the long sought, highly electrophilic diethylaluminum moiety in Et(2)Al(CB(11)H(6)X(6)). X-ray crystallography reveals ion-like structural features reminiscent of the corresponding trialkylsilylium species. Et(2)Al(CB(11)H(6)X(6)) is a potent catalyst for the electrophilic ethenation of benzene, the polymerization of cyclohexene oxide, and the oligomerization of ethene to a low molecular weight, highly branched product. PMID- 12083911 TI - Diuranium inverted sandwiches involving naphthalene and cyclooctatetraene. AB - Treatment of IU(DME)(NC[(t)Bu]Mes)(3) (2-I-DME) with 4 equiv of KC(8) and 0.5 equiv of naphthalene in DME allowed the isolation of a naphthalene-bridged compound, K(2)(mu-eta(6),eta(6)-C(10)H(8))[U(NC[(t)Bu]Mes)(3)](2) (K(2)-2(2)-mu C(10)H(8)), in 60% yield as a dark brown powder. The twelve U-C distances are rather short, varying from 2.565(11) to 2.749(10) A. Treatment of M(2)-2(2)-mu C(10)H(8) (M = Na, K) with 2 equiv of 1,3,5,7-cyclooctatetraene afforded a mixture of two products: M-2-COT and 2(2)-mu-COT. Compound 2(2)-mu-COT can be assembled independently in 90% yield by salt elimination upon reaction of M-2-COT with iodide 2-I-DME. The U-C(arene) distance in compound 2(2)-mu-COT is longer than that in its naphthalene counterpart K(2)-2(2)-mu-C(10)H(8)(2.822 vs 2.634 A), in accord with bonding considerations. A DFT study performed on model compounds for both M(2)-2(2)-mu-C(10)H(8) and 2(2)-mu-COT indicates that the delta bonds present in the former compound show better covalent overlap. PMID- 12083913 TI - New poly(phenyleneethynylene)s with cationic, facially amphiphilic structures. AB - Polymers based on meta substituted phenylene ethylene are prepared with patterned polar and nonpolar groups to favor an extended conformation. These polymers were characterized at the air-water interface by Langmuir techniques and found to form stable monolayers with an extended conformation based on molecular models. In addition, these polymers show phospholipid membrane activity as measured by induced leakage of calcein from large unilamellar vesicles. These polymers represent new facially amphiphilic structures which are cationic in nature and surface active. PMID- 12083914 TI - [(18)O]-oxygen incorporation reveals novel pathways in spiroacetal biosynthesis by Bactrocera cacuminata and B. cucumis. AB - The origins of the oxygen atoms in 1,7-dioxaspiro[5.5]undecane (1) and hydroxyspiroacetal (2) from Bactrocera cacuminata, and in 2,8-dimethyl-1,7 dioxaspiro[5.5]undecane (3) and hydroxyspiroacetal (4) from B. cucumis, have been investigated by incorporation studies from both [(18)O(2)]-dioxygen and [(18)O] water. Combined GC-MS examination and high-field NMR analysis have demonstrated that all oxygen atoms in 1 and 2 from B. cacuminata are dioxygen derived, but in contrast, the spiroacetals 3 and 4 from B. cucumis incorporate one ring oxygen from water and one ring oxygen (and the hydroxyl oxygen in 4) from [(18)O(2)] dioxygen. These results reveal not only the generality of monoxygenase mediation of spiroacetal formation in Bactrocera sp., but also an unexpected complexity in their biosynthesis. A general paradigm accommodating these and other observations is presented. PMID- 12083915 TI - Photonic switching of photoinduced electron transfer in a dithienylethene porphyrin-fullerene triad molecule. AB - A dithienylethene (DTE)-porphyrin (P)-fullerene (C(60)) triad molecule in which intramolecular photoinduced electron transfer is controlled by the photochromic DTE moiety has been prepared. Irradiation of the molecule with visible light gives the open form of the dithienylethene (DTEo). Excitation of the porphyrin gives DTEo-(1)P-C(60), which undergoes photoinduced electron transfer with a time constant of 25 ps to generate DTEo-P(.+)-C(60)(.-). Irradiation with ultraviolet light produces the closed form of the dithienylethene (DTEc). Excitation of DTEc P-C(60) yields DTEc-(1)P-C(60), whose porphyrin first excited singlet state is quenched in 2.3 ps by singlet-singlet energy transfer to DTEc, generating (1)DTEc P-C(60) and precluding significant photoinduced electron transfer. Such highly reversible photonically controlled intramolecular photoinduced electron transfer may eventually be useful in the design of photonic or optoelectronic devices. PMID- 12083916 TI - Characterization of a didehydrobenzoxazine intermediate in a novel carbene-to carbene transformation. AB - Irradiation of N(2) matrix-isolated 3-chloro-3-(2-benzoxazolyl)diazirine gives a mixture of syn- and anti-benzoxazolylchlorocarbenes which could be characterized by IR, UV/vis, and B3LYP modeling. Subsequent irradiation of the carbene caused ring opening to the corresponding quinoimine, which was similarly characterized. In turn, the quinoimine photochemically underwent ring-closure to a novel, highly strained cyclic ketenimine. Unrestricted singlet DFT calculations were required to fit the IR spectrum of the ketenimine, suggesting significant diradical character. Photolysis of the ketenimine led to ring cleavage in yet another fashion, to give an isonitrile-phenoxychlorocarbene, whose spectra were similar to those of the previously characterized phenoxychlorocarbene. Finally, at shorter wavelengths, the phenoxycarbene underwent the expected rearrangement to the corresponding benzoyl chloride. PMID- 12083917 TI - Hydrogen uptake in boron nitride nanotubes at room temperature. AB - Hydrogen uptake capacities of 1.8 and 2.6 wt % were obtained on BN multiwalled nanotubes and bamboo-like nanotubes, respectively, under approximately 10 MPa at room temperature. The preliminary results show that chemical interactions mainly take place between hydrogen and BN. Taking into consideration the uniform semiconducting electronic property and ultimate stable chemical and thermal stability, BN nanotubes may exhibit some interesting possibilities in storing hydrogen. This will also provide some viewpoints on designing the carbon-based hydrogen storage system. PMID- 12083918 TI - First isolation and characterization of an anti-apicophilic spirophosphorane bearing an oxaphosphetane ring: a model for the possible reactive intermediate in the Wittig reaction. AB - An anti-apicophilic phosphorane bearing an oxaphosphetane ring, in which the ring carbon is apical and the ring oxygen is equatorial (C-apical), has been prepared as a thermally less stable stereoisomer of a phosphorane with an ordinary equatorial carbon-apical oxygen array in the oxaphosphetane ring (O-apical). This novel C-apical phosphorane, which could be considered to be a model compound of the reactive intermediate in the Wittig reaction, was fully characterized by NMR and X-ray structural analysis. The compound was found to easily isomerize to its more stable O-apical isomer, especially in the presence of proton sources, and the latter O-apical compound was found to furnish olefin at elevated temperatures. PMID- 12083919 TI - Photolysis of gamma-(alpha-carboxy-2-nitrobenzyl)-L-glutamic acid investigated in the mcrosecond time scale by time-resolved FTIR. AB - The photolytic release of substrates from caged substrates has proven to be an excellent method to generate concentration jumps for kinetic measurements in the microsecond time scale. In this report we use time-resolved FTIR in the step-scan mode to probe the photolysis mechanism of one such caged compound, namely gamma (alpha-carboxy-2-nitrobenzyl)glutamate, and to obtain a direct measure of the rate of photorelease of the substrate glutamate. The time-resolved difference FTIR spectra exhibit specific signals that can be assigned to the reactant caged glutamate, photolytically released product glutamate, as well as to the aci-nitro intermediate, the key intermediate of the photolysis reaction. Therefore these signals allow the characterization of the kinetics of formation and decay of the intermediate and products. This is the first such report that provides a direct determination of the rate of formation of the photolysis product from a caged compound in the microsecond time scale. Furthermore, the results presented provide a good basis for further time-resolved FTIR studies of molecular reaction mechanisms, such as ligand protein interactions, in the microsecond time scale through the photolytic release of substrates from caged compounds. PMID- 12083920 TI - A novel peptide-based encoding system for "one-bead one-compound" peptidomimetic and small molecule combinatorial libraries. AB - The "one-bead one-compound" (OBOC) combinatorial library method is highly efficient, especially when used with well-established on-bead binding or functional assays. Literally, millions of compounds can be screened concurrently within 1 to 2 days. However, structure determination of peptidomimetic and small molecule compounds on one single bead is not trivial. A novel, highly efficient, and robust peptide-based encoding system has been developed for OBOC peptidomimetic and small molecule combinatorial libraries. In this system, topologically segregated bifunctional beads, which are made by a simple biphasic solvent strategy, are employed for the preparation and screening of an OBOC combinatorial peptidomimetic and small molecule libraries. Testing molecules are on the outer layer, and the coding tags in the interior of the bead do not interfere with screening. The coding tag is a peptide containing a large number of unnatural alpha-amino acids derived from different building blocks used for generating the peptidomimetic or small molecule. By coupling common building blocks simultaneously to the scaffold of the testing compound and to the side chains of the alpha-amino acids on the coding peptide, extra synthetic steps are eliminated and the amount of undesirable side products is minimized. Positive bead decoding is easy and straightforward as there is no need for cleavage and retrieval of the coding tag, and positive beads can be sequenced directly with Edman degradation. To demonstrate the efficiency and simplicity of our encoding system, an encoded 158 400-member model peptidomimetic library has been generated and screened for ligands that bind to streptavidin. Potent and novel ligands with clear motifs have been identified. PMID- 12083921 TI - Pentalenene synthase. Analysis of active site residues by site-directed mutagenesis. AB - Incubation of farnesyl diphosphate (1) with the W308F or W308F/H309F mutants of pentalenene synthase, an enzyme from Streptomyces UC5319, yielded pentalenene (2), accompanied by varying proportions of (+)-germacrene A (7) with relatively minor changes in k(cat) and k(cat)/K(m). By contrast, single H309 mutants gave rise to both (+)-germacrene A (7) and protoilludene (8) in addition to pentalenene (2). Mutation to glutamate of each of the three aspartate residues in the Mg(2+)-binding aspartate-rich domain, (80)DDLFD, resulted in reduction in the k(cat)/K(m) for farnesyl diphosphate and formation of varying proportions of pentalenene and (+)-germacrene A (7). Formation of (+)-germacrene A (7) by the various pentalenene synthase mutants is the result of a derailment of the natural anti-Markovnikov cyclization reaction, and not simply the consequence of trapping of a normally cryptic, carbocationic intermediate. Both the N219A and N219L mutants of pentalenene synthase were completely inactive, while the corresponding N219D mutant had a k(cat)/K(m) which was 3300-fold lower than that of the wild type synthase, and produced a mixture of pentalenene (2) (91%) and the aberrant cyclization product beta-caryophyllene (9) (9%). Finally, the F77Y mutant had a k(cat)/K(m) which was reduced by 20-fold compared to that of the wild-type synthase. PMID- 12083922 TI - Electrostatic properties of membrane lipids coupled to metarhodopsin II formation in visual transduction. AB - Changes in lipid composition have recently been shown to exert appreciable influences on the activities of membrane-bound proteins and peptides. We tested the hypothesis that the conformational states of rhodopsin linked to visual signal transduction are related to biophysical properties of the membrane lipid bilayer. For bovine rhodopsin, the meta I-meta II conformational transition was studied in egg phosphatidylcholine (PC) recombinants versus the native rod outer segment (ROS) membranes by means of flash photolysis. Formation of metarhodopsin II was observed by the change in absorbance at 478 nm after a single actinic flash was delivered to the sample. The meta I/meta II ratio was investigated as a function of both temperature and pH. The data clearly demonstrated thermodynamic reversibility of the transition for both the egg PC recombinants and the native ROS membranes. A significant shift of the apparent pK(a) for the acid-base equilibrium to lower values was evident in the egg PC recombinant, with little meta II produced under physiological conditions. Calculations of the membrane surface pH using a Poisson-Boltzmann model suggested the free energies of the meta I and meta II states were significantly affected by electrostatic properties of the bilayer lipids. In the ROS membranes, phosphatidylserine (PS) is needed for full formation of meta II, in combination with phosphatidylethanolamine (PE) and polyunsaturated docosahexaenoic acid (DHA; 22:6omega3) chains. We propose that the PS surface potential leads to an accumulation of hydronium ions, H(3)O(+), in the electrical double layer, which drive the reaction together with the large negative spontaneous curvature (H(0)) conferred by PE plus DHA chains. The elastic stress/strain of the bilayer arises from an interplay of the approximately zero H(0) from PS and the negative H(0) due to the PE headgroups and polyunsaturated chains. The lipid influences are further explained in terms of matching of the bilayer spontaneous curvature to the curvature at the lipid/rhodopsin interface, as formulated by the Helfrich bending energy. These new findings guide current ideas as to how bilayer properties govern the conformational energetics of integral membrane proteins. Moreover, they yield knowledge of how membrane lipid-protein interactions involving acidic phospholipids such as PS and neutral polyunsaturated DHA chains are implicated in key biological functions such as vision. PMID- 12083923 TI - High-throughput NMR-based screening with competition binding experiments. AB - The Achilles heel of ligand-based NMR screening methods is their failure to detect high-affinity ligands and molecules that bind covalently to the receptor. We have developed a novel approach for performing high-throughput screening with NMR spectroscopy that overcomes this limitation. The method also permits detection of potential high-affinity molecules that are only marginally soluble, thus significantly enlarging the diversity of compounds amenable to NMR screening. The techniques developed utilize transverse and/or selective longitudinal relaxation parameters in combination with competition binding experiments. Mathematical expressions are derived for proper setup of the NMR experiments and for extracting an approximate value of the binding constant for the identified ligand from a single-point measurement. With this approach it is possible to screen thousands of compounds in a short period of time against protein or DNA and RNA fragments. The methodology can also be applied for screening plant and fungi extracts. PMID- 12083925 TI - Molecular "compasses" and "gyroscopes." III. Dynamics of a phenylene rotor and clathrated benzene in a slipping-gear crystal lattice. AB - Samples of 1,4-bis(3,3,3-triphenylpropynyl)benzene 3 were prepared by Pd(0) catalyzed coupling of 3,3,3-triphenylpropyne (1) and 1,4-diiodobenzene. The structure of compound 3 is such that the central phenylene can play the role of a gyroscope wheel, while the alkyne bond and trityl groups can act as an axle and shielding frameworks, respectively. Crystals grown from benzene and dichloromethane were characterized by X-ray diffraction, variable-temperature (13)C CPMAS NMR, quadrupolar echo solid-state (2)H NMR, and thermal analyses. The rotational dynamics of benzene molecules and phenylene groups were characterized in terms of 6-fold rotation and 2-fold flipping models, respectively. The possibility of a gearing mechanism between adjacent benzene molecules and phenylene groups suggested by the clathrate structure was investigated. However, it was found that 6-fold rotation of benzene molecules at 300 K occurs in the gigahertz regime (or higher) and 2-fold flipping of phenylene groups in the kilohertz range in a structure that can be described as a slipping-gear lattice. The rotational dynamics of the phenylene group in the solvent-free structure were remarkably similar to those in the clathrate, and both are among the fastest known for phenylene rotation in solids. The results presented here provide a valuable starting point for the design and analysis of crystalline solids with correlated molecular motions. PMID- 12083924 TI - Structural control in thin layers of poly(p-phenyleneethynylene)s: photophysical studies of Langmuir and Langmuir-Blodgett films. AB - We present the relationship between the spatial arrangement and the photophysical properties of fluorescent polymers in thin films with controlled structures. Eight surfactant poly(p-phenyleneethynylene)s were designed and studied. These detailed studies of the behavior of the polymers at the air-water interface, and of the photophysical properties of their transferred LB films, revealed key structure-property relationships. Some of the polymers displayed pi-aggregates that are characteristic of an edge-on structure at the air-water interface. Monolayer LB films of these polymers showed greatly reduced quantum yields relative to solution values. Other polymers exhibited a highly emissive face-on structure at the air-water interface, and did not form pi-aggregates. The combination of pressure-area isotherms and the surface pressure dependent in situ UV-vis spectra of the polymers at the air-water interface revealed different behavioral details. In addition, the UV-vis spectra, fluorescence spectra, and quantum yields of the LB films provide design principles for making highly emissive films. PMID- 12083926 TI - Stable dialkyl ether/poly(hydrogen fluoride) complexes: dimethyl ether/poly(hydrogen fluoride), a new, convenient, and effective fluorinating agent. AB - The preparation, (1)H, (13)C, and (19)F NMR structural characterization as well as with DFT-based theoretical calculations of stable dialkyl ether/poly(hydrogen fluoride) complexes are reported. Dimethyl ether/poly(hydrogen fluoride) (DMEPHF), are stable complexes of particular interest and use. The DFT calculations, that are in agreement with NMR data, suggest a cyclic poly(hydrogen fluoride) bridged structure for DMEPHF. The complex, DME-5 HF was found to be a convenient and effective new fluorinating agent with the ease of workup and applied to several fluorination reactions, such as the hydrofluorination and bromofluorination of alkenes, and fluorination of alcohols giving good to excellent yield with high selectivity. Homologous dialkyl ether/poly(hydrogen fluoride) (R(2)O/[HF](n,), R = Et, nPr) systems are also stable and suitable for fluorination reactions. PMID- 12083927 TI - Solvation phenomena of a tetrapeptide in water/trifluoroethanol and water/ethanol mixtures: a diffusion NMR, intermolecular NOE, and molecular dynamics study. AB - Solvation of a tetrapeptide, NAc-Ser-Phe-Val-Gly-OMe (1), in water and in water/alcohol mixtures with 2,2,2-trifluoroethanol (TFE)/water or ethanol (ETH)/water has been studied by diffusion NMR and intermolecular NOE measurements. The experimental results were compared with those obtained from detailed Molecular Dynamics (MD) calculations. Independently, all three methods revealed preferential solvation on the surface of the peptide by TFE in the water/TFE mixtures, but not by ETH in the water/ETH mixtures. The MD calculations show that the TFE concentration coating the peptide is higher than that in the bulk, while for ethanol, the concentration is nearly equal to that in the bulk. Calculated site-specific preferential solvation data between TFE, ETH, and water with the different peptide groups have been compared with the NMR data and shown to be in general agreement with the experimental facts. PMID- 12083929 TI - The tightness contribution to the Bronsted alpha for hydride transfer between NAD+ analogues. AB - It has been shown that the rate of symmetrical hydride transfer reaction varies with the hydride affinity of the (identical) donor and acceptor. In that case, Marcus theory of atom and group transfer predicts that the Bronsted alpha depends on the location of the substituent, whether it is in the donor or the acceptor, and the tightness of the critical configuration, as well as the resemblance of the critical configuration to reactants or products. This prediction has now been confirmed for hydride transfer reactions between heterocyclic, nitrogen containing cations, which can be regarded as analogues of the enzyme cofactor, nicotinamide adenine dinucleotide (NAD+). A series of reactions with substituents in the donor gives Bronsted alpha of 0.67 +/- 0.03 and a tightness parameter, tau, of 0.64 +/- 0.06. With substituents in the acceptor alpha = 0.32 +/- 0.03 and tau = 0.68 +/- 0.08. The reactions are all spontaneous, with equilibrium constants between 0.4 and 3 x 10(4), and the two sets span about the same range of equilibrium constants. The two tau values are essentially identical with an average value of 0.66 +/- 0.05. These results can be semiquantitatively mimicked by rate constants calculated for a linear, triatomic model of the reaction. Variational transition state theory and a physically motivated but empirically calibrated potential function were used. The computed rate constants generate an alpha value of 0.56 if the hydride affinity of the acceptor is varied and an alpha of 0.44 if the hydride affinity of the donor is varied. The calculated kinetic isotope effects are similar to the measured values. A previous error in the Born charging term of the potential function has been corrected. Marcus theory can be successfully fitted to both the experimental and computed rate constants, and appears to be the most compact way to express and compare them. The success of the linear triatomic model in qualitatively reproducing these results encourages the continued use of this easily conceptualized model to think about group, ion, and atom transfer reactions. PMID- 12083930 TI - Functionalizable polycyclic aromatics through oxidative cyclization of pendant thiophenes. AB - We present a general strategy for obtaining large sulfur-containing polycyclic aromatics from thienyl precursors through iron(III) chloride mediated oxidative cyclizations. By placing thienyl moieties in close proximity to adjacent arenes, we have directed the oxidized intermediates into controlled cyclization pathways, effectively suppressing polymer formation. Utilizing these cyclized compounds and their thienyl precursors, we have studied cyclization/polymerization pathways of polymers such as poly(2). The unsubstituted positions alpha to the sulfur atoms within these aromatic cores allowed for efficient halogenation and further functionalization. As a demonstration, we prepared a series of arylene-ethynylene polymers with varying degrees of chromophore aromatization and used them to probe the effects of synthetically imposed rigidity on polymer photophysical behavior. The symmetries and effective conjugation pathways within the monomers play a key role in determining photophysical properties. We observed that rigid, aromatized chromophores generally led to increased excited-state lifetimes by decreasing radiative rates of fluorescence decay. PMID- 12083928 TI - Formation of isoprostane bicyclic endoperoxides from the autoxidation of cholesteryl arachidonate. AB - Autoxidation of polyunsaturated fatty acids and esters leads to a complex mixture containing hydroperoxides and cyclic peroxides. Prostaglandin bicyclic endoperoxides have been detected from the autoxidation of cholesteryl arachidonate by LC-MS and GC-MS techniques. All four possible types (I-IV) of bicyclic endoperoxides have been found starting from different regioisomeric hydroperoxides of cholesteryl arachidonate. Furthermore, the stereochemistry of Type IV bicyclic endoperoxides has been determined by conversion to pentafluorobenzyl (PFB) ester, trimethylsilyl (TMS) derivatives, and comparison with synthetic standards by the use of GC-MS. All eight possible diastereomers of the derivatized isoprostanes were observed and were separated by gas chromatography. The bicyclic endoperoxides with the two alkyl chains syn on the cyclopentane ring were formed preferentially to those with anti configuration, a result anticipated from earlier work. Substantial amounts of the anti-substituted isoprostanes, including PGF(2)(alpha), were, however, observed in the product mixture. PMID- 12083931 TI - Combined solid state NMR and X-ray diffraction investigation of the local structure of the five-coordinate silicon in fluoride-containing as-synthesized STF zeolite. AB - The local structure of the [SiO(4/2)F]- unit in fluoride-containing as synthesized STF zeolite has been experimentally determined by a combination of solid-state NMR and microcrystal X-ray diffraction to be very close to trigonal bipyramidal. Because the fluoride ions are disordered over two sites, the resulting local structure of the [SiO(4/2)F]- unit from a conventional XRD refinement is an average between tetrahedral SiO(4/2) and five-coordinate [[SiO(4/2)F]-, giving an apparent F-Si distance longer than expected. The correct F-Si distance was determined by slow spinning MAS and fast spinning (19)F/(29)Si CP and REDOR solid-state NMR experiments and found to be between 1.72 and 1.79 A. In light of this, the X-ray structure was re-refined, including the disorder at Si3. The resulting local structure of the [SiO(4/2)F]- unit was very close to trigonal bipyramidal with a F-Si distance of 1.744 (6) A, in agreement with the NMR results and the prediction of Density Functional Theory calculations. In addition, further evidence for the existence of a covalent F-Si bond is provided by a (19)F-->(29)Si refocused INEPT experiment. The resonance for the five coordinate species at -147.5 ppm in the (29)Si spectrum is a doublet due to the (19)F/(29)Si J-coupling of 165 Hz. The peaks in this doublet have remarkably different effective chemical shift anisotropies due to the interplay of the CSA, dipolar coupling, and J-coupling tensors. The distortions from tetrahedral geometry of the neighboring silicon atoms to the five-coordinate Si3 atom are manifested in increased delta(aniso) values. This information, along with F-Si distances measured by (19)F-->(29)Si CP experiments, makes it possible to assign half of the (29)Si resonances to unique tetrahedral sites. As well as determining the local geometry of the [SiO(4/2)F]- unit, the work presented here demonstrates the complementarity of the solid-state NMR and X-ray diffraction techniques and the advantages of using them together. PMID- 12083932 TI - Luminescent lanthanide complexes of a bis-bipyridine-phosphine-oxide ligand as tools for anion detection. AB - The Gd(3+), Tb(3+), and Eu(3+) complexes of a bis-bipyridine-phenylphosphine oxide ligand PhP(O)(bipy)(2) 1 (bipy for 6-methylene-6'-methyl-2,2'-bipyridine) have been synthesized. In acetonitrile solutions at room temperature, the Tb(3+) and Eu(3+) complexes show a metal-centered luminescence, indicative of an efficient energy transfer from the two bipy subunits to the Ln center. The photophysical properties drastically depend on the nature of the anions present in solution. In particular, addition of 2 equiv of nitrate anions to a solution containing the [Ln.1](OTf-)(3) leads to an 11-fold increase of the luminescence intensity for the Eu(3+) and a 7-fold increase for the Tb(3+) complexes. Similar effects are provided with Cl-, F-, and CH(3)COO- anions. UV-vis titration experiments were used to determine association constants for binding of, respectively, one, two, and three anions. Stepwise anion addition has also been investigated on the molecular level using quantum mechanical (QM) calculations for the Eu complexes. These calculations reproduce the experimental findings, especially if solvent molecule addition is taken into account. The X-ray crystal structure of the nitrate salt of the Tb complex, as well as QM calculation of a similar Eu complex, demonstrates the coordination of three nitrate anions in a bidentate mode and the step-by-step relegation of the bipy subunits in the second coordination sphere. These features give valuable insights into the mechanism of the overall light amplification process. PMID- 12083933 TI - Highly selective addition of chiral, sulfonimidoyl substituted bis(allyl)titanium complexes to N-sulfonyl alpha-imino esters: asymmetric synthesis of gamma,delta unsaturated alpha-amino acids bearing a chiral, electron-withdrawing nucleofuge at the delta-position. AB - Selective addition of the chiral, sulfonimidoyl substituted bis(allyl)titanium complexes 5a-d, which are configurationally labile in regard to the Calpha-atoms, to N-toluenesulfonyl (Ts)-, N-2-trimethylsilylethanesulfonyl (SES)-, and N-tert butylsulfonyl (Bus) alpha-imino ester (9a-c) in the presence of Ti(OiPr)(4) and ClTi(OiPr)(3) afforded with high regio- and diastereoselectivities in good yields the (syn, E)-configured beta-alkyl-gamma,delta-unsaturated alpha-amino acid derivatives 2a-g, which carry a chiral, electron-withdrawing nucleofuge at the delta-position and a cyclohexyl, an isopropyl, a phenyl, and a methyl group at the beta-position. Addition of the cyclic bis(allyl)titanium complex 14 to N-Bus alpha-imino ester 9c afforded with similar high regio- and diastereoselectivities the (E)- and (Z)-configured amino acid derivatives (E)-8 and (Z)-8. Reaction of complexes 5a-d with alpha-imino esters 9a-c in the presence of Ti(OiPr)(4) occurs stepwise to give first the mono(allyl)titanium complexes containing 2a-g as ligands, which react in the presence of ClTi(OiPr)(3) with a second molecule of 9a-c with formation of two molecules of 2a-g. Formation of (S,R,E)-configured homoallylic amines 2a-g entails Si,Re,E processes of alpha-imino esters 9a-c with the (R,R)-configured bis(allyl)titanium complexes (R,R)-5a-d and (R)-configured mono(allyl)titanium complexes (R)-17a-d, both of which are most likely in rapid equilibrium with their (S,S)-diastereomers and (S)-diastereomers, respectively. Interestingly, in the reaction of 5a-d with aldehydes, the (S,S)-configured complexes (S,S)-5a-d are the ones which react faster. Reaction of the N-titanated amino acid derivatives Ti-2a and Ti-2b with N-Ts alpha-imino ester 9a led to the highly diastereoselective formation of imidazolidinones 15a and 15b, respectively. Cleavage of the sulfonamide group of the N-Bus amino acid derivative 2d with CF(3)SO(3)H gave quantitatively the sulfonimidoyl functionalized amino acid H-2d. A Ni-catalyzed cross-coupling reaction of the amino acid derivative 2e with ZnPh(2) led to a substitution of the sulfonimidoyl group by a phenyl group and furnished the enantiomerically pure protected alpha amino acid Bus-1. Two new N-sulfonyl alpha-imino esters, the SES and the Bus alpha-imino esters 9b and 9c, respectively, have been synthesized from the corresponding sulfonamides by the Kresze method in medium to good yields. The N SES alpha-imino ester 9b and the N-Bus alpha-imino ester 9c should find many synthetic applications, in particular, in cases where the N-Ts alpha-imino ester 9a had been used before. PMID- 12083934 TI - Long-range electron transfer across molecule-nanocrystalline semiconductor interfaces using tripodal sensitizers. AB - Four tripodal sensitizers, Ru(bpy)(2)(Ad-tripod-phen)(2+) (1), Ru(bpy)(2)(Ad tripod-bpy)(2+) (2), Ru(bpy)(2)(C-tripod-phen)(2+) (3), and Ru(bpy)(2)(C-tripod bpy)(2+) (4) (where bpy is 2,2'-bipyridine, phen is 1,10-phenanthroline, and Ad tripod-bpy (phen) and C-tripod-bpy (phen) are tripod-shaped bpy (phen) ligands based on 1,3,5,7-tetraphenyladamantane and tetraphenylmethane, respectively), have been synthesized and characterized. The tripodal sensitizers consist of a rigid-rod arm linked to a Ru(II)-polypyridine complex at one end and three COOR groups on the other end that bind to metal oxide nanoparticle surfaces. The excited-state and redox properties of solvated and surface-bound 1-4 have been studied at room temperature. The absorption spectra, emission spectra, and electrochemical properties of 1-4 in acetonitrile solution are preserved when 1-4 are bound to nanocrystalline (anatase) TiO(2) or colloidal ZrO(2) mesoporous films. This behavior is indicative of weak electronic coupling between TiO(2) and the sensitizer. The kinetics for excited-state decay are exponential for 1-4 in solution and are nonexponential when 1-4 are bound to ZrO(2) or TiO(2). Efficient and rapid (k(cs) > 10(8) s(-)(1)) excited-state electron injection is observed for 1-4/TiO(2). The recombination of the injected electron with the oxidized Ru(III) center is well described by a second-order kinetic model with rate constants that are independent of the sensitizer. The sensitizers bound to TiO(2) were reversibly oxidized electrochemically with an apparent diffusion coefficient approximately 1 x 10(-11) cm(2) s(-)(1). PMID- 12083935 TI - Open-framework and microporous vanadium silicates. AB - A series of novel vanadium silicates with open-framework and microporous structures has been synthesized under mild hydrothermal conditions. Ten distinct framework types have been identified that all have structures based on cross linking single silicate sheets with square pyramidal V(IV)O(5) units to give compounds with the general formula A(r)[(VO)(s)(Si(2)O(5))(p)(SiO(2))(q)].tH(2)O, where A is Na, K, Rb, Cs, or a combination. The vanadosilicate (VSH-n) structures have free channel diameters up to 6.5 A and show good thermal stability, absorption, and ion-exchange properties, suggesting their potential for technological applications as molecular sieves or in catalysis. PMID- 12083936 TI - Location of protons in anhydrous Keggin heteropolyacids H(3)PMo(12)O(40) and H(3)PW(12)O(40) by (1)H[(31)P]/(31)P[(1)H] REDOR NMR and DFT quantum chemical calculations. AB - HeteroPolyAcids (HPA's) are a class of solid acids that have broad applications in many fields of science and technology, including catalysis and chemical engineering. The proton locations within the thermally stable and commonly known Keggin unit, which is the primary structure building unit/block, has remained undetermined in anhydrous HPAs, despite numerous theoretical and experimental efforts. However, Rotational Echo DOuble Resonance (REDOR) NMR and Density Functional Theory (DFT) quantum chemical calculations offer a new opportunity to determine the exact locations of protons within the Keggin unit. The crucial experimental evidence is provided for the basic and very extensively studied acidic form of H(8-n)X(n+)M(12)O(40), X = Si, P and M = Mo, W, belonging to the Keggin structure. While showing that the acidic protons are located in the bridging oxygen positions (R(P-H) = 520 +/- 20 pm) in H(3)PMo(12)O(40) and in the terminal oxygen positions (R(P-H) = 570 +/- 20 pm) in H(3)PW(12)O(40), REDOR measurements also provide for the first time the structural basis to consistently rank the acid strength for the important class of Keggin solid catalysts. PMID- 12083937 TI - (57)Fe Mossbauer isomer shifts of heme protein model systems: electronic structure calculations. AB - We report the results of density functional theory (DFT) calculations of the (57)Fe Mossbauer isomer shifts (delta(Fe)) for a series of 24 inorganic, organometallic, and metalloprotein/metalloporphyrin model systems in S = 0, (1)/(2), 1, (3)/(2), 2, and (5)/(2) spin states. We find an excellent correlation between calculation and experiment over the entire 2.34 mm s(-1) range of isomer shifts: a 0.07-0.08 mm s(-1) rms deviation between calculation and experiment (corresponding to 3-4% of the total delta(Fe) range, depending on the functionals used) with R(2) values of 0.973 and 0.981 (p < 0.0001). The best results are obtained by using the hybrid exchange-correlation functional B3LYP, used previously for (57)Fe Mossbauer quadrupole splittings and (57)Fe NMR chemical shifts and chemical shielding anisotropies. The relativistically corrected value of alpha, alpha(rel), converges with the large basis set used in this work, but the exact values vary somewhat with the methods used: -0.253 a(0)(3) mm s(-1) (Hartree-Fock; HF); -0.316 a(0)(3) mm s(-1) (hybrid HF-DFT; B3LYP), or -0.367 a(0)(3) mm s(-1) (pure DFT; BPW91). Both normal and intermediate spin state isomer shifts are well reproduced by the calculations, as is the broad range of delta(Fe) values: from [Fe(VI)O(4)](2-) (-0.90 mm s(-1) expt; -1.01 mm s(-1) calc) to KFe(II)F(3) (1.44 mm s(-1) expt; 1.46 mm s(-1) calc). Molecular orbital analyses of all inorganic solids as well as all organometallic and metalloporphyrin systems studied reveal that there are three major core MO contributions to rho(tot)(0), the total charge density at the iron nucleus (and hence delta(Fe)), that do not vary with changes in chemistry, while the valence MO contributions are highly correlated with delta(Fe) (R(2) = 0.915-0.938, depending on the functionals used), and the correlation between the valence MO contributions and the total MO contribution is even better (R(2) = 0.965-0.976, depending on the functionals used). These results are of general interest since they demonstrate that DFT methods now enable the accurate prediction of delta(Fe) values in inorganic, organometallic, and metalloporphyrin systems in all spin states and over a very wide range of delta(Fe) values with a very small rms error. PMID- 12083938 TI - Development of a molecular recognition ion gating membrane and estimation of its pore size control. AB - We have fabricated a molecular recognition ion gating membrane. This synthetic membrane spontaneously opens and closes its pores in response to specific solvated ions. In addition to this switching function, we found that this membrane could control its pore size in response to a known concentration of a specific ion. The membrane was prepared by plasma graft copolymerization, which filled the pores of porous polyethylene film with a copolymer of NIPAM (N isopropylacrylamide) and BCAm (benzo[18]crown-6-acrylamide). NIPAM is well-known to have an LCST (lower critical solution temperature), at which its volume changes dramatically in water. The crown receptor of the BCAm traps a specific ion, and causes a shift in the LCST. Therefore, selectively responding to either K(+) or Ba(2+), the grafted copolymer swelled and shrank in the pores at a constant temperature between two LCSTs. The solution flux in the absence of Ba(2+) decreased by about 2 orders of magnitude over a solution flux containing Ba(2+). The pore size was estimated by the filtration of aqueous dextran solutions with various solute sizes. This revealed that the membrane changed its pore size between 5 and 27 nm in response to the Ba(2+) concentration changes. No such change was observed for Ca(2+) solutions. Furthermore, this pore size change occurred uniformly in all pores, as a clear cut-off value for a solute size that could pass through pores was always present. This membrane may be useful not only as a molecular recognition ion gate, but also as a device for spontaneously controlling the permeation flux and solute size. PMID- 12083939 TI - Asymmetric total synthesis of 11-deoxytetrodotoxin, a naturally occurring congener. AB - Tetrodotoxin, a toxic principle of puffer fish poisoning, is a specific blocker of sodium channel. Despite many synthetic efforts since the structure elucidation in 1964, the only total synthesis of the racemic tetrodotoxin has been reported by Kishi and co-workers. In the course of our studies directed toward the total synthesis to analyze biologically interesting issues associated with tetrodotoxin, we accomplished a highly stereocontrolled synthesis of (-)-5,11 dideoxytetrodotoxin in 1999. Based on the synthesis, we describe herein the first total synthesis of 11-deoxytetrodotoxin, a naturally occurring analogue. The synthesis started from an allylic alcohol, the same intermediate for the synthesis of 5,11-dideoxytetrodotoxin. Epoxidation of the allylic alcohol was followed by isomerization with Ti(i-PrO)(4) to give an alpha-hydroxy allylic alcohol, in which the configurations of the two hydroxyl groups were inverted by oxidation and then a 2-step reduction. Further epoxidation of the allylic alcohol and ozonolysis of the remaining vinyl group gave an aldehyde, which reacted with magnesium acetylide to give a propargyl alcohol in a stereoselective manner. Oxidative cleavage of the acetylenic moiety with RuO(4) afforded a fully functionalized lactone for 11-deoxytetrodotoxin. Crucial guanidinylation was achieved from trichloroacetamide according to our own method to give acetyldibenzylguanidine. Finally, deprotection of benzyl groups, acetates, and acetal furnished 11-deoxytetrodotoxin. PMID- 12083940 TI - Self-assembly does not account for the hydrophobic effect. AB - Marmur has claimed that large values of activity coefficients for nonelectrolytes, particularly in the context of hydrophobic interactions between solutes in aqueous solution at ambient temperature and pressure, cannot be accounted for by thermodynamics, and has suggested that association (self assembly) of solute molecules in solution solves this dilemma. We show that the analysis of Marmur is incorrect, specifically because the equilibrium in solution between monomeric solute molecules and associated solute molecules is entirely ignored. We show further that activity coefficients such as that for nitromethane solute in hexane solvent, 39.7, and that for solute hexane in solvent water, 4.48 x 10(5), can be calculated as 31.9 and 4.71 x 10(5), respectively, by methods based on well-known molecule-molecule interactions. No assumption of self assembly is required. PMID- 12083941 TI - Understanding the role of stereoelectronic effects in determining collagen stability. 2. A quantum mechanical/molecular mechanical study of (Proline-Proline Glycine)(n) polypeptides. AB - The importance of vicinal and long-range interresidue effects in determining the stability of the collagen triple helix has been investigated by quantum mechanical (QM) and molecular mechanical (MM) computations on suitable model polypeptides, taking into account solvent effects by the polarizable continuum model (PCM). At the QM level, the PII conformation corresponds to an energy minimum for pentapeptide analogues incorporating the sequence Gly-Pro-Pro-Gly, irrespective of the down or up puckering of the pyrrolidine ring. However, our computations indicate that the alternation of down and up prolines characterizing collagen and collagen-like peptides is not due to an intrinsic preference of the Pro-Pro-Gly sequence. This result is confirmed by MM computations of longer polypeptides. Next, MM computations on model triple helices show that a better packing is obtained for specific values of backbone dihedrals, which, in turn, favor the alternation of down and up prolines along each chain. PMID- 12083942 TI - Toward the spectrum of free polyethylene: linear alkanes studied by carbon 1s photoelectron spectroscopy and theory. AB - Trends in carbon 1s ionization energies for the linear alkanes have been investigated using third-generation synchrotron radiation. The study comprises CH(4), C(2)H(6), C(3)H(8), C(4)H(10), C(5)H(12), C(6)H(14), and C(8)H(18). Both inter- and intramolecular shifts in ionization energy have been determined from gas-phase spectra and ab initio calculations. The shifts are decomposed into initial-state and final-state contributions and are shown to relate to the fundamental chemical properties of group electronegativity and polarizability. By extrapolation, we predict C1s spectra of larger n-alkanes, converging toward isolated strands of polyethylene. PMID- 12083943 TI - Clinical importance of beta-adrenoceptor polymorphisms in cardiovascular disease. AB - beta-Adrenoceptor antagonists play an important role in the treatment of cardiovascular disease and have been used for three decades in the treatment of hypertension and ischemic heart disease. More recently they have been demonstrated to improve survival in patients with mild to moderate congestive heart failure. The beneficial effects of beta-adrenoceptor antagonists stems from their ability to limit the deleterious effects of adrenergic stimulation, which in the cardiovascular system is primarily transmitted through two subclasses of receptor, beta(1) and beta(2). The advances of the Human Genome Project have led to an increased appreciation that variations in genetic background may underlie a substantial portion of the clinical heterogeneity apparent in cardiovascular disease. This review examines the molecular, functional, and clinical significance of the most common polymorphisms of the beta(1 and beta(2) adrenoceptors. Initial research in adrenoceptor variation focused on the beta(2) adrenoceptor. Three common polymorphisms appear to influence receptor function: Arg16-->Gly, Glu(27)-->Gln, and Thr(164)-->Ile. In in vitro studies of agonist stimulation, Gly(16) receptors demonstrate enhanced downregulation, while Glu(27) variants are resistant to downregulation. There is much controversy and conflict among various clinical studies regarding the effect of these variants on vasoreactivity and hypertensive risk. The Ile(164) variant demonstrates decreased responsiveness to agonist activity both in vitro and in animal models. In studies of patients with congestive heart failure, this variant has been associated with poor functional capacity and decreased survival. More recent investigations have focused on the two common polymorphisms of the beta(1)-adrenoceptor: Ser(49)- >Gly, and Arg(389)-->Gly. In vitro studies of Arg(389) receptors demonstrate a gain of function, as agonist stimulation results in significantly higher intracellular levels of cyclic adenosine monophosphate when compared with the Gly(389) variant. Consistent with the in vitro data, clinical studies demonstrate increased responsiveness to beta-agonist stimulation, and an increased risk of hypertension among Arg(389) homozygotes. Further investigation of the clinical implications of these common variants of beta(1)- and beta2)-adrenoceptors are needed. Importantly, the pharmacogenetic impact of these variants on the effectiveness of beta-adrenergic blockade remains unknown. PMID- 12083944 TI - Metabolic enzyme polymorphisms and susceptibility to acute leukemia in adults. AB - Genetic approaches to understanding the etiology of the acute leukemias are beginning to deliver meaningful insights. Polymorphic variants in xenobiotic metabolizer loci were a natural starting point to study the relevance of these changes. The finding that glutathione S-transferase (GST) T1 null variants increase leukemia risk has implicated oxidative stress in hematopoietic stem cells as an important etiological factor in acute myeloid leukemia (AML). The importance of these enzyme systems in handling specific substrates has also been confirmed by the finding of an increased risk of therapy-related leukemia in individuals with underactive variants of GSTP1 who have been exposed to a chemotherapeutic agent metabolized by this enzyme. Benzene is a well-recognized leukemogen, and genetic variants in its metabolic pathway can modulate the risk of leukemia following exposure. In particular, underactive variants of the NAD(P)H:quinone oxidoreductase 1 gene (NQO1) seem to increase the risk of AML. Other enzymes within the pathway are proving more difficult to study because of the absence of variants that significantly affect the biological activity of the enzyme under study. No effect of the myeloperoxidase (MPO) gene variants in altering the risk of AML has been seen in our studies. Another pathway recently shown to be important in determining leukemia risk is folic acid metabolism, particularly important in predisposition to acute lymphocytic leukemia (ALL). Polymorphic variants of the methylenetetrahydrofolate reductase gene (MTHFR) which impair its activity have been shown to be associated with a protective effect. This is thought to be due to an increased availability of nucleotide precursors for incorporation into DNA. This finding implicates misincorporation of uracil into DNA as an important mechanism of leukemic change in lymphoid precursors. Future studies will extend these observations but will require biological material collected from large well-controlled epidemiological studies. The technological challenges imposed by the high throughput of samples required by these studies are currently being addressed. PMID- 12083945 TI - Genetic characterization of growth hormone deficiency and resistance: implications for treatment with recombinant growth hormone. AB - Growth failure can be caused by deficient growth hormone production or action. The genes involved in pituitary development, somatotrope function, as well as growth hormone synthesis, secretion, and action have recently been characterized in considerable detail. Familial growth failure has played an important role in identifying these genes, and a large number of mutations adversely affecting the development and function of the growth hormone/insulin-like growth factor axis have been discovered. Inactivating mutations leading to growth retardation in humans have been identified in several pituitary transcription factor genes (HESX1, PITX2, LHX3, PROP1, POU1F1) as well as in genes encoding the growth hormone-releasing hormone receptor (GHRH-R), the G(s) protein alpha subunit (GNAS1), growth hormone itself (GH-1), the growth hormone receptor (GHR), and in a single case each, the insulin-like growth factor I (IGF-I) and the IGF-I receptor. Mutations in pituitary transcription factors cause developmental abnormalities of the pituitary and deficiency of multiple pituitary hormones [growth hormone (GH), prolactin (Prl), thyrotropin (TSH) and lutropin/follitropin (LH/FSH)]. Most of the syndromes respond well to therapy with recombinant GH; exceptions are antibody-mediated resistance in GHD type IA (not all patients) and cases of Laron syndrome (GHR deficiency). Such patients respond to IGF-I therapy. This review summarizes the molecular genetics, functional defects, phenotypes, diagnostic considerations and therapeutic aspects of syndromes associated with mutations in the relevant genes. PMID- 12083946 TI - Functional genomics approaches for the identification and validation of antifungal drug targets. AB - So far, antifungal drug discovery seems to have benefited little from the enormous advances in the field of genomics in the last decade. Although it has become clear that traditional drug screening is not delivering the long-awaited novel potent antifungals, little has been reported on efforts to use novel genome based methodologies in the quest for new drugs acting on human pathogenic fungi. Although the market for a novel systemic and even topical broad-spectrum antifungal appears considerable, many large pharmaceutical companies have decided to scale back their activities in antifungal drug discovery. Here we report on some of the recent advances in genomics-based technologies that will allow us not only to identify and validate novel drug targets but hopefully also to discover active therapeutic agents. Novel drug targets have already been found by 'en masse' gene inactivation strategies (e.g. using antisense RNA inhibition). In addition, genome expression profiling using DNA microarrays helps to assign gene function but also to understand better the mechanism of action of known drugs (e.g. itraconazole) and to elucidate how new drug candidates work. No doubt, we have a long way to go just to catch up with the advances made in other therapeutic areas, but all tools are at hand to derive practical benefits from the genomics revolution. The next few years should prove a very exciting time in the history of antifungal drug discovery. PMID- 12083947 TI - Preventing type 1 diabetes mellitus: the promise of gene therapy. AB - Type 1 (insulin-dependent) diabetes mellitus is an autoimmune disease that has no cure. Closed-loop insulin administration strategies and approaches for replacement of the insulin-producing beta cells may offer improved treatments, which could delay or prevent diabetes complications. In the long run, however, prevention of type 1 diabetes in susceptible individuals represents the best chance for reducing the toll of the disease. Prevention of type 1 diabetes will require reliable methods for early diagnosis of predisposition to the disease, using improved genetic and serological screening on a wide scale. Identification of the primary antigenic target(s) for autoimmunity will allow intervention in prediabetes stages aimed at the induction of antigen-specific tolerance. In addition to manipulation of the immune system, the susceptibility of beta cells to autoimmunity could be reduced. A number of genes have been shown to increase beta-cell resistance to immune effector molecules in animal models and cultured beta-cell lines. These genes could be used for preventive gene therapy of type 1 diabetes mellitus if expressed in beta cells prior to the onset of autoimmune destruction. This prospect depends on the development of safe and efficient vectors, and approaches for cell-specific targeting of these vectors to beta cells in vivo. PMID- 12083949 TI - Influence of GSTT1 and GSTM1 genotypes on sunburn sensitivity. AB - BACKGROUND: Exposure to sunlight may cause sunburn, skin cancer or phototoxic reactions to certain drugs such as Hypericum extract. All these are ultraviolet B (UVB)-mediated reactions which may be modulated by individual genetic susceptibility. UVB exposure results in oxidative stress. Many products of oxidative stress are detoxified by glutathione-S-transferases mu 1 (GSTM1) and theta 1 (GSTT1). Deletion polymorphisms (genotype *0/*0) of GSTM1 and GSTT1 occur in 50% and 20% of Caucasians, respectively. By affecting the individual ability to detoxify oxidative stress-related products, they may influence the severity of the cutaneous photoreaction. METHODS: Minimal erythema doses (MED) of UVB irradiation on the skin were determined in 110 subjects who were selected according to their GSTT1 genotype (28 GSTT1*0/*0, 54 GSTT1*A/*0, and 28 GSTT1*A/*A). Genotypes were detected with novel polymerase chain reaction (PCR) assays that allow the differentiation between homozygous and heterozygous GSTT1 and GSTM1 deletions. RESULTS: In the absence of GSTT1 enzyme, the susceptibility of individuals to UVB-induced inflammatory skin reactions increased significantly (p = 0.02, ANCOVA). 'Gene-equivalents' were calculated from the number of functional GSTM1 and GSTT1 alleles as a measure of the gene-dose. UVB sensitivity correlated with gene dose up to a threshold above which additional GSTT1 or GSTM1 alleles did not provide additional protection. Volunteers who were homozygously deficient in GSTT1 and GSTM1 were most sensitive to UVB. Interestingly, individuals with high GSTM1 gene-doses showed increased photosensitization after administration of Hypericum extract (St. John's wort). CONCLUSION: Individuals harboring the *0/*0 genotype of GSTT1 and/or GSTM1 showed enhanced UVB-induced cutaneous damage. Moreover, GST genotypes modulated Hypericum-induced photosensitization. PMID- 12083948 TI - Gene therapy for Parkinson's disease: current status and future potential. AB - Parkinson's disease appears to be a good candidate for gene therapy. The primary biochemical defect associated with the disease has been clearly determined as an absence of dopamine in the caudate-putamen, and the anatomical region where the neuropathologic hallmark of the disease, death of the nigral dopamine-producing neurons, occurs, remains circumscribed. Based on the biochemical and anatomical information gathered on Parkinson's disease, different gene therapy strategies have been devised to treat it. The first, and most explored strategy so far, consists in engineering cells to produce levodopa or dopamine so they will replace dopaminergic neurotransmission. Several types of cells have been employed in these experiments, and behavioral recovery has been reported in animal models of the disease. However, this approach cannot prevent neuronal death, nor reconstruct brain circuits. Another strategy is to protect cells by transferring genes that encode neurotrophic factors. Effort is now being concentrated into this research area, and promising results have recently been reported. Finally, an additional strategy aims at generating cells with a dopaminergic phenotype so they will be capable of replacing the missing dopaminergic neurons in biochemical, anatomical and functional terms. This has the potential to become an important constituent for an effective cure. Gene therapy holds significant promise for the treatment of neurodegenerative disorders, and Parkinson's disease treatment will benefit greatly from the knowledge and information arising from gene therapy research. PMID- 12083950 TI - Genome screening for susceptibility loci in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a complex, multigenic autoimmune disease with a wide spectrum of clinical manifestations. Much of the pathology is attributed to deposition to various tissues of immune complexes continuously formed with autoantibodies; thus, the pathogenesis is related to dysregulation of self-reactive B cells. Recent family linkage studies and allele-sharing linkage analyses of affected sibling pairs have advanced genome screening for susceptibility loci in SLE, and a considerable number of chromosomal intervals with significant or suggestive linkage to SLE have been identified. However, there are still several inherent difficulties in precisely identifying loci and genes, as the complexity of polygenic inheritance of SLE phenotypes is considerable. One must note that each specific aspect of diverse SLE phenotypes (clinical manifestations and immunological abnormalities) is mostly controlled separately by a different set of susceptibility loci. Involvement of positive and negative epistatic gene interactions often puzzles genetic analyses. Studies on SLE using murine lupus models are ongoing to solve some of these difficulties. Comparative studies have identified several syntenic chromosomal intervals with susceptibility loci in both mouse models and humans. Thus, combining knowledge derived from both human and murine studies is vital. The ultimate identification of susceptibility genes and their functions will probably depend largely on studies using genetically manipulated mutant mice, including those with homologous recombination of potent polymorphic target genes. The up-coming completion of genomic sequences in mice and humans is predicted to limit the numbers of potent candidate genes in particular genomic intervals and accelerates this line of studies. Such knowledge will lead to elucidation of genetic and cellular mechanisms involved in the dysregulation of self-reactive lymphocytes in the pathogenesis of SLE. Prophylactic and therapeutic clinical approaches can then be better designed. PMID- 12083951 TI - Polymorphisms in immunoregulatory genes: towards individualized immunosuppressive therapy? AB - In organ transplantation, successful immunosuppression requires that both rejection and infection episodes be minimized. Unfortunately it is currently impossible to predict individual dose requirement for immunosuppressive drugs, but a number of studies of various immune response genes are now being performed with a view to identifying genotypes associated with rejection and/or infection. The key role of cytokines in the immune response and other processes, including fibrosis, has concentrated most of this attention on polymorphisms in cytokine genes. Data on polymorphisms in genes encoding tumor necrosis factor-alpha, transforming growth factor-beta, interferon-gamma and interleukin (IL)-1, 4, 6 and 10 together with the IL-4 receptor have been analyzed but so far there is currently no indication of any consistently positive associations between graft rejection and any of these polymorphisms. Studies of other immunomodulatory genes including the CTLA4 gene and the chemokine receptor CCR-5 have proved more positive though the data, so far, are only preliminary. In conclusion, additional large series studies of these and other cytokine genes, as well as other immunoregulatory gene polymorphisms of proven functional significance are needed to achieve major progress in this area. PMID- 12083952 TI - Mobile genetic element activation and genotoxic cancer therapy: potential clinical implications. AB - Approximately one-quarter of the human genome is composed of short and long interspersed elements (SINEs and LINEs, respectively). These elements have spread throughout the genome by a process termed retrotransposition, consisting of transcription of an element into RNA, reverse transcription into cDNA, and reinsertion of the copied element into a new genomic location. Recombination events involving these elements, including novel insertions into active genes, have been associated with a number of human diseases. Despite the fact that these elements have replicated to hundreds of thousands of copies in the genome, under most conditions they remain transcriptionally silent, and therefore are not actively replicating. The signals controlling retrotransposable element activation in the genome have not been defined. Our laboratory recently found that exposure of cells to a variety of DNA-damaging agents, including several common chemotherapeutic drugs and gamma-radiation, is associated with dramatic induction of SINE transcription, and of a concomitant endogenous reverse transcriptase activity. As SINEs do not encode for reverse transcriptase, the latter finding suggests a more global activation of retrotransposable elements in response to DNA damage. Together these observations suggest that genotoxic exposure may lead to genomic mutation not only through direct DNA damage, but also through indirect activation of potentially mutagenic mobile elements in the genome. The nonrandom distribution of retrotransposable elements in the human genome may contribute to the pattern of characteristic translocation events associated with secondary malignancies in patients exposed to genotoxic agents. Here we describe these and other mechanisms by which retrotransposable elements can contribute to disease, and present an overview of what is known about this large, and largely unexplored, segment of the genome. Understanding the cellular responses to genotoxic stress may permit the development of a means of predicting the risks and preventing the development of secondary malignancy following cancer therapy. PMID- 12083953 TI - Genetics of multiple sclerosis: linkage and association studies. AB - Multiple sclerosis (MS) is a demyelinating autoimmune disease of the central nervous system caused by an interplay of environmental and genetic factors. The only genetic region that has been clearly demonstrated by linkage and association studies to contribute to MS genetic susceptibility is the human leukocyte antigen (HLA) system. The majority of HLA population studies in MS have focused on Caucasians of Northern European descent, where the predisposition to disease has been consistently associated with the class II DRB1*1501-DQA1*0102-DQB1*0602 haplotype. A positive association with DR4 was detected in Sardinians and in other Mediterranean populations. Moreover DR1, DR7, DR11 have been found to be protective in several populations. Systematic searches aimed at identifying non HLA susceptibility genes were undertaken in several populations by means of linkage studies with microsatellite markers distributed across the whole genome. The conclusion of these studies was that there is no major MS locus, and genetic susceptibility to the disease is most likely explained by the presence of different genes each conferring a small contribution to the overall familial aggregation. The involvement of several candidate genes was tested by association studies, utilizing either a population-based (case control) or a family-based (transmission disequilibrium test) approach. Candidate genes were selected mainly on the basis of their involvement in the autoimmune pathogenesis and include immunorelevant molecules such as cytokines, cytokine receptors, immunoglobulin, T cell receptor subunits and myelin antigens. With the notable exception of HLA, association studies met only modest success. This failure may result from the small size of the tested samples and the small number of markers considered for each gene. New tools for large scale screening are needed to identify genetic determinants with a low phenotypic effect. Large collaborative studies are planned to screen several thousands of patients with MS with several thousands of genetic markers. The tests are increasingly based on the DNA pooling procedure. PMID- 12083954 TI - Bacterial proteomics and vaccine development. AB - Until recently, the development of vaccines for use in humans relied on the response to attenuated or whole-cell preparations, or empirically selected antigens. The post-genomic era holds the possibility of rational design of novel vaccines for important human pathogens. The discovery and development of these new vaccines is likely to be accomplished through integrated proteomic strategies. Although most proteomic studies are based on two-dimensional gel electrophoresis (2D-PAGE) as a separation technique, new methods have been developed within the past two years that provide complementary information concerning microbial protein expression. The 2D-PAGE technique in combination with Western blotting has been successfully applied in the discovery of antigens from Helicobacter pylori, Chlamydia trachomatis and Borrelia garinii. Two dimensional semi-preparative electrophoresis has provided complementary information regarding membrane protein expression in a strain of H. pylori. Through two-dimensional liquid chromatography-tandem mass spectrometry, the most comprehensive information to date regarding protein expression in yeast was obtained. This technique may shortly become an important tool in vaccinology. This review of the current state of bacterial proteomics as applied in vaccinology presents analytical techniques for protein separation, proteomics without gels, reverse vaccinology, and functional approaches to the identification of virulence proteins in microbes. PMID- 12083955 TI - Histamine genomics in silico: polymorphisms of the human genes involved in the synthesis, action and degradation of histamine. AB - BACKGROUND: Histamine is a ubiquitous biogenic amine involved in the regulation of numerous basic physiological and pathophysiological processes. The DNA sequences of the genes encoding proteins (enzymes and receptors) that participate in the synthesis, degradation and cellular binding of histamine are already identified. OBJECTIVE: We analyzed the in silico available human sequences to find genetic polymorphisms in histamine-related genes (L-histidine decarboxylase, histamine receptors, histamine N-methyl transferase and diamine-oxidase), and compared these data with findings concerning structure-function relationships in order to get information about the possible pathophysiological relevance of these polymorphisms. METHODS: Sequence analysis was performed at the National Center for Biotechnology Information Database. The search tool BLAST was applied. RESULTS: Several sequence variations were found, and it is conceivable that some of these genetic polymorphisms may be related to various pathological conditions. Among sequence variations, variants with no amino acid change, variants resulting in amino acid alterations, and many nucleotide changes involving non-coding sequences were revealed. CONCLUSIONS: Histamine genomics may provide a new tool for medical prediction and drug design in the future. PMID- 12083956 TI - Androgen receptor mutations in carcinoma of the prostate: significance for endocrine therapy. AB - Endocrine therapy for advanced prostate cancer involves androgen ablation (orchiectomy or application of luteinizing hormone releasing hormone analogs) and/or blockade of the androgen receptor (AR) with either steroidal (cyproterone acetate) or nonsteroidal (hydroxyflutamide, bicalutamide and nilutamide) antiandrogens. These antagonists prevent androgen-induced conformational change and activation of the AR. During long term androgen ablation, the AR adapts to an environment with low androgen concentrations and becomes hypersensitive to low concentrations of androgens, either alone or in combination with various cellular regulators. Bicalutamide can switch from antagonist to agonist during long-term androgen withdrawal, as shown in prostate cancer LNCaP cells. AR point mutations were detected in metastatic lesions from human prostate cancer more frequently than in primary tumors. Although functional characterization of only some mutant AR detected in prostate cancer tissue has been performed, data available suggest that they are activated by dihydrotestosterone, its precursors and metabolites, synthetic androgens, estrogenic and progestagenic steroids and hydroxyflutamide. A direct association between AR mutations and endocrine withdrawal syndrome has been investigated in only one study thus far. There is no evidence at present that activation of any of the mutant AR genes detected in prostate cancer is enhanced in the presence of a nonsteroidal AR stimulator. Coactivators of the AR are proteins that associate with the receptor, possess histone acetylase activity and facilitate AR activation. The coregulatory proteins ARA70 and ARA160 differentially affected the activity of the mutated AR Glu(231)-->Gly, which was discovered in a mouse authochthonous prostate tumor. ARA70 enhanced receptor activation by both androgen and estradiol, whereas ARA160 augmented only androgen induced AR activity. Novel experimental therapies that down-regulate AR expression have been developed; they include the application of ribozymes and antisense oligonucleotides. PMID- 12083957 TI - Candidate genes for atopic asthma: current results from genome screens. AB - Atopic asthma is one of the most common childhood diseases in developed countries. Asthma is characterized by reversible airway obstruction, bronchial hyper-responsiveness, and airway inflammation. Atopy in childhood is considered the strongest predisposing factor for asthma. The etiology of asthma is complex and is thought to involve the interaction of multiple genes and a variety of environmental factors such as allergens and viral and bacterial infections. To identify genes conferring susceptibility for asthma and atopy, many genome-wide screens for asthma and its associated traits have now been carried out, and genetic linkage has been consistently identified in several regions. Several independent genome-wide screens found regions of linkage with asthma on chromosomes 5, 6, 11, 12, 13, 16 and 19, identifying candidate susceptibility genes including FCER1B, the IL-4 gene cluster, TNFalpha, HLA loci and others. However, the evidence for linkage is still only suggestive for most regions. In an effort to clarify the mechanism underlying the development of asthma, further studies utilizing new technologies and data from the Human Genome Project are ongoing. It is hoped that this accumulation of data will lead to improved genetic testing and assist in the development of new drugs. PMID- 12083958 TI - Bacterial genomics as a potential tool for discovering new antimicrobial agents. AB - The past 30 years have witnessed the emergence of new infectious diseases as well as the re-emergence of those thought to be defeated or under control. It is likely that this threat will continue and that infectious micro-organisms will be found to be responsible for numerous diseases whose etiology had been previously unknown. Compounding this threat is the rapid evolution of drug resistance by micro-organisms that is rendering many existing antimicrobial agents obsolete. Thus, there is an urgent need for the development of new classes of antimicrobial agents and the identification of new drug targets. Over the past decade, advances in high-throughput automated DNA sequencing have delivered a wealth of genetic information in the form of whole genome sequences of microbial pathogens. Coupled with this advancement has been the development of new genetic tools and computational advances capable of selecting genes of particular interest as well as testing for the effects of candidate drugs. While no new drugs have yet been developed, further study into the application and limitations of these new approaches to the identification of novel targets will aid in overcoming the current problem of antimicrobial drug resistance. PMID- 12083959 TI - Drug target pharmacogenomics: an overview. AB - Pharmacogenomics is a field aimed at understanding the genetic contribution to variability in drug efficacy and toxicity. The goal is to be able to select the drugs with the greatest likelihood of benefit and the least likelihood of harm in individual patients, based on their genetic make-up. Pharmacogenetics has historically been a field focused primarily on genetic polymorphisms in drug metabolizing enzymes and their impact on drug efficacy and toxicity. More recently, investigators have begun to study the relationship between drug target polymorphisms and drug efficacy and toxicity. There are now numerous examples in the literature of associations between drug target polymorphisms and drug effect. Drug targets can be broken into three main categories: the direct protein target of the drug, signal transduction cascades or downstream proteins, and disease pathogenesis proteins. While the drug target pharmacogenetics literature provides 'proof of concept' that genetic variability contributes to the variability in drug response, the data are not to the point of being clinically useful in most cases. Specific problems to date include the inability of a single polymorphism to be highly predictive of response, and inconsistencies across studies of the same polymorphism. It seems likely that an important factor in the above limitations is the approach of focusing on a single polymorphism in a single gene. Given that most drug responses involve a large number of proteins, all of whose genes could have several polymorphisms, it seems unlikely that a single polymorphism in a single gene would explain a high degree of drug response variability in a consistent fashion. Thus, it seems that a polygenic, or genomic approach will be more appropriate. Candidate gene and genome scanning approaches to pharmacogenomics have shown promise in relating drug target polymorphisms to response or toxicity, and pharmacogenomic strategies for drug discovery and drug development are now being implemented by most major pharmaceutical companies. Pharmacogenomics has the potential to significantly enhance the ability of clinicians to use medications in a safe and effective manner and, as such, represents an exciting field with tremendous clinical potential. PMID- 12083960 TI - Detecting rare mutations associated with cancer risk. AB - For more than a decade, investigators have been searching for a means of determining the risk of individuals developing cancer by detecting rare oncogenic mutations. The accumulation of mutations and the clonal evolvement of tumors provide opportunities for monitoring disease development and intervening prior to the presentation of clinical symptoms, or determining the risk of disease relapse during remission. A number of techniques, mostly polymerase chain reaction (PCR) based, have been developed that enable the detection of rare oncogenic mutations within the range of 10(-2) to 10(-4) wild-type cells. Only a handful of procedures enable the detection of intragenic single base mutations at one mutant in 10-6 or better. These ultra-sensitive mutation detection techniques have produced some interesting results regarding single base mutation spectra and frequencies in p53, Harvey-ras, N-ras, and other reporter genes and DNA sequences in human tissues. Although there is evidence that some individuals may harbor cells or clones expressing genomic instability, the connection with the processes of carcinogenesis is still tenuous. There remains a need for rigorous epidemiological studies employing these ultra-sensitive mutation detection procedures. Since genomic instability is considered key to tumor development, the relevance of the detection of hypermutable clones in individuals is discussed in the context of cancer risk. PMID- 12083961 TI - High throughput genotyping technologies for pharmacogenomics. AB - Genetic differences between individuals play a role in determining susceptibility to diseases as well as in drug response. The challenge today is first to discover the range of genetic variability in the human population and then to define the particular gene variants, or alleles, that contribute to clinically important outcomes. Consequently, high throughput, automated methods are being developed that allow rapid scoring of microsatellite alleles and single nucleotide polymorphisms (SNPs). Many detection technologies are being used to accomplish this goal, including electrophoresis, standard fluorescence, fluorescence polarization, fluorescence resonance energy transfer, and mass spectrometry. SNP alleles may be distinguished by any one of several methods, including single nucleotide primer extension, allele-specific hybridization, allele-specific primer extension, oligonucleotide ligation assay, and invasive signal amplification. Newer methods require less sample manipulation, increase sensitivity, allow more flexibility, and decrease reagent costs. Recent developments show promise for continuing these trends by combining amplification and detection steps and providing flexible, miniaturized platforms for genotyping. PMID- 12083962 TI - Information technology tools for efficient SNP studies. AB - We are currently facing a new era of studies involving single nucleotide polymorphisms (SNPs). This increased attention is stimulated by interest in individual differences in disease susceptibility as well as individual responses to drug treatment and the falling cost of genotyping. This review is a guide to the numerous public data repositories and Information Technology (IT) tools that may aid planning, preparation, running and analysis of studies involving SNPs. I will also highlight areas where researchers will have to resort to home-made IT solutions. Unfortunately, both information and IT tools are scattered throughout the internet and a lack of data exchange conventions can hamper the efficient use of these existing resources. This can lead to situations where the planning, preparation and analysis of a SNP study can actually cost more than the actual genotyping. We propose that only a customizable backbone IT infrastructure for SNP studies can help reduce costs associated with SNP data handling and tool launching. PMID- 12083963 TI - Determination of human beta(2)-adrenoceptor haplotypes by denaturation selective amplification and subtractive genotyping. AB - OBJECTIVE: beta(2)-Adrenoceptor haplotype may be better associated with asthma severity and drug response than a polymorphic variant at any single site. Because present methods of haplotype determination are time consuming and impractical for large population studies, we sought to develop a simple and efficient method of determining haplotype of 3 common polymorphisms at codons -19 (Arg/Cys), 16 (Arg/Gly) and 27 (Glu/Gln). DESIGN: Preliminary studies showed that the C/G base pair of the Arg(-19) allele increases the local melting temperature over the T/A base pair of the Cys(-19) allele by 3.6 degrees C and establishes a new local maximum denaturation temperature. By choosing a suitable denaturation temperature and appropriate primers and coupling them with restriction fragment length polymorphism (RFLP) analysis, we hypothesized that the genotype of one separately amplified allele followed by subtraction from the combined genotype of two alleles would yield the beta(2) haplotype in > 99% of the population. RESULTS: Haplotype determined by our method was in complete agreement with haplotype determined by cloning and sequencing in 29 samples. The frequencies of haplotype pairs in 78 healthy adults, according to our method, were in agreement with published values that were inferred, and were: RGE/CRQ, 26.9%; CRQ/CRQ, 25.6%; RGE/CGQ, 16.7%; CRQ/CGQ, 10.3%; RGE/RGE, 11.5%; CGQ/CGQ, 7.7%. The haplotype pair in one individual was RRE/CRQ (1.3%). CONCLUSION: Our method of determining beta(2)-adrenoceptor haplotype is simple, accurate and cost effective for haplotyping large populations. PMID- 12083964 TI - Polymorphism of the serotonin transporter: implications for the use of selective serotonin reuptake inhibitors. AB - Selective serotonin reuptake inhibitors (SSRIs) are used to treat a number of psychiatric disorders related to mood and anxiety, and variations in the serotonin transporter (5-HTT) gene may be involved in a number of these. A polymorphic site in the promoter region is associated with differences in 5-HTT gene expression. Studies suggest that the short allele of the 5-HTT promoter (5 HTTPR) site can adversely influence the antidepressant response to SSRIs, and is associated with anxiety-related traits, depression, and impulsive disorders such as alcohol abuse. Several studies do not replicate these findings; potential confounding factors include age, gender, and population stratification. Other 5 HTT polymorphisms also exist. For example, individuals with the short allele of a variable number of tandem repeats (VNTR) polymorphism, located in the second intron, may have reduced responsiveness to SSRIs, and the STin2.12 allele at this site has been associated with bipolar disorder. Findings both supporting and inconsistent with these conclusions are reviewed. The clinical effects of the polymorphisms may be associated with effects on platelets, neural 5-HTT levels, and indices of serotonergic function. PMID- 12083965 TI - Genetic variation of the beta(2)-adrenoceptor: its functional and clinical importance in bronchial asthma. AB - Asthma is a polygenic disease for which no clear genotype-phenotype relationships have emerged. In contrast, although not associated with the diagnosis of asthma per se, variant forms of the beta(2)-adrenoceptor (beta2-AR) gene (ADRB2) display functional effects that may be clinically relevant. Single nucleotide polymorphisms (SNPs) of ADBR2 are common and result in amino acid substitutions at positions 16, 27, and 164 of the receptor as well as position 19 of its 5' upstream peptide. These SNPs influence receptor function in vitro, although evidence regarding exact relationships is conflicting. This has raised the possibility that phenotypes such as bronchial hyper-responsiveness (BHR) and responses to (beta2)-agonist drugs may be genetically determined. To date, no unequivocal relationships between SNPs and phenotype have been identified. In some studies the Gly(16) allele has been associated with increased BHR and asthma severity. In others, the Arg(16) allele has been shown to determine acute bronchodilator response and adverse events during long term beta(2)-agonist therapy. The latter may provide the basis for clinical application of this new knowledge. More recently, a small number of frequently occurring, functionally relevant ADRB2 haplotype pairs have been confirmed. These combinations of alleles may be more important in determining genotype/phenotype relationships than individual SNPs, and may explain why earlier investigations have yielded contrasting results. Future studies will be required to clarify the pharmacodynamic effects of ADRB2haplotypes both in vitro and in vivo. PMID- 12083966 TI - The molecular genetics of bone formation: implications for therapeutic interventions in bone disorders. AB - Skeletal biology is a complex process involving the developmental commitment and differentiation of chondrocytes and osteoblasts which produce and mineralize cartilage and bone matrix during growth and postnatal life. Several genes are involved in controlling osteogenesis by acting on target cells in a very complex manner. Manipulation of genes in mice and studies of genetic mutations affecting the skeleton in humans have enabled the assessment of the role of transcription factors, bone matrix proteins and regulatory factors involved in the control of chondrocyte and osteoblast differentiation, and have considerably improved our understanding of the bone formation process. Clinical studies and gene polymorphism analyses suggest that the variable expression of particular genes may be linked to clinical osteoporosis. A major challenge in the future will be to develop molecularly targeted approaches to stimulating bone formation and increasing bone mass. The use of mouse strain models and transgenic animals with variable bone density may be useful to identify genetic determinants of bone mass which may serve as a basis for drug discovery and development. On the other hand, the availability of gene microarrays and other emerging genomic techniques are promising tools to identify genes that are distinctly expressed in health and disease. These technologies may also serve to test the mechanisms of action of drugs aimed at increasing bone formation. Genetic studies of the molecular signaling pathways involved in normal and pathological osteogenesis may also help to identify genes that could be targeted for therapeutic intervention. Candidate approaches include selective gene transfection in target cells and the use of drugs acting on gene promoters to selectively enhance gene expression in osteoblasts. The development of these strategies is expected not only to bring new insight into the molecular mechanisms that govern bone formation in normal and pathological situations but, in the long term, may also result in the identification of novel molecular targets for therapeutic interventions for bone formation disorders. PMID- 12083967 TI - Polymorphisms in the methylenetetrahydrofolate reductase gene: clinical consequences. AB - 5,10-Methylenetetrahydrofolate reductase (MTHFR) plays a key role in folate metabolism by channeling one-carbon units between nucleotide synthesis and methylation reactions. Severe enzyme deficiency leads to hyperhomocysteinemia and homocystinuria, with altered folate distribution and a phenotype that is characterized by damage to the nervous and vascular systems. Two frequent polymorphisms in the human MTHFR gene confer moderate functional impairment of MTHFR activity for homozygous mutant individuals. The C to T change at nucleotide position 677, whose functional consequences are dependent on folate status, has been extensively studied for its clinical consequences. A second polymorphism, an A to C change at nucleotide position 1298, is not as well characterized. Still equivocal are associations between MTHFR polymorphisms and vascular arteriosclerotic or thrombotic disease. Neural tube defects and pregnancy complications appear to be linked to impaired MTHFR function. Colonic cancer and acute leukemia, however, appear to be less frequent in individuals homozygous for the 677T polymorphism.MTHFR polymorphisms influence the homocysteine-lowering effect of folates and could modify the pharmacodynamics of antifolates and many other drugs whose metabolism, biochemical effects, or target structures require methylation reactions. However, only preliminary evidence exists for gene-drug interactions. This review summarizes the biochemical basis and clinical evidence for interactions between MTHFR polymorphisms and several disease entities, as well as potential interactions with drug therapies. Future investigations of MTHFR in disease should consider the influence of other variants of functionally related genes as well as the medication regimen of the patients. Animal models for genetic deficiencies in folate metabolism will likely play a greater role in our understanding of folate-dependent disorders. PMID- 12083968 TI - Finding genes influencing susceptibility to complex diseases in the post-genome era. AB - During the last decade, hundreds of genes that harbor mutations causing simple Mendelian disorders have been identified using a combination of linkage analysis and positional cloning techniques. Traditional approaches to gene mapping have been largely unsuccessful in mapping genes influencing so-called 'complex' genetic diseases, however, because of low power and other factors. Complex genetic diseases do not display simple Mendelian patterns of inheritance, although genes do have an influence and close relatives of probands consequently have an increased risk. These disorders are thought to be due to the combined effects of variation at multiple interacting genes and the environment. Complex diseases have a significant impact on human health because of their high population incidence (unlike simple Mendelian disorders, which tend to be rare). New techniques are being developed aimed specifically at mapping genes conferring susceptibility to complex diseases. A project aimed at mapping genes influencing susceptibility to a complex disease may be undertaken in several stages: establishing a genetic basis for the disease in one or more populations; measuring the distribution of gene effects; studying statistical power using models; carrying out marker-based mapping studies using linkage or association. Quantitative genetic models can be used to estimate the heritability of a complex (polygenic) disease, as well as to predict the distribution of gene effects and to test whether one or more quantitative trait loci (QTLs) exist. Such models can be used to predict the power of different mapping approaches, but are often unrealistic and therefore provide only approximate predictions. Linkage analyses, association studies and family-based association tests are all hindered by low power and other specific problems. Association studies tend to be more powerful but can generate spurious associations due to population admixture. Alternative strategies for association mapping include the use of recent founder populations or unique isolated populations that are genetically homogeneous, and the use of unlinked markers (so-called genomic controls) to assign different regions of the genome of an admixed individual to particular source populations. Linkage disequilibrium observed in a sample of unrelated affected and normal individuals can also be used to fine-map a disease susceptibility locus in a candidate region. New Bayesian strategies make use of an annotated human genome sequence to further refine the position of a candidate disease susceptibility locus. PMID- 12083969 TI - Cystic fibrosis and the use of pharmacogenomics to determine surrogate endpoints for drug discovery. AB - Cystic fibrosis (CF) is caused by a mutation in the CFTR gene, encoding a chloride channel. For the most common mutation, Delta F508, the basis of the deficit is the failure of the mutant CFTR channel protein to traffic properly to the apical plasma membrane of the affected epithelial cell. The trafficking failure results in loss of the cyclic adenosine monophosphate (cAMP)-activated chloride channel function of the CFTR protein in the plasma membrane. The lung is the principal site affecting patient morbidity and mortality in CF. The main reason is that the CF airway epithelial cells also secrete high levels of the proinflammatory cytokine interleukin (IL)-8, resulting in massive cellular inflammation, infection, tissue damage and lung destruction. The relationship between the trafficking defect, the loss of chloride channel activity, and inflammation is not known. However, gene therapy of CF lung epithelial cells with the wild-type CFTR gene can repair the chloride channel defect, as well as suppress the intrinsic hypersecretion of IL-8. Repair of both defective channels and high IL-8 secretion can also be effected by treatment with the candidate CF drug CPX, which is in clinical trials in CF patients. CPX acts by binding to the mutant CFTR protein, and helps the protein to mature and gain access to the plasma membrane. CPX also suppresses the synthesis and secretion of IL-8 from CF epithelial cells, presumably by virtue of its repair of the trafficking defect of mutant CFTR. To guide pharmacogenomic experiments we have therefore hypothesized that the genomic signature of CF epithelial cells treated with CPX should resemble the signature of the same cells repaired by gene therapy. We have developed two algorithms for identifying genes modified by repair of CFTR defects. The GRASP algorithm uses a statistical test to identify the most profoundly changing genes. The GENESAVER algorithm allows us to identify those genes whose pattern of expression changes in-phase or out-of-phase with IL-8 secretion by CF cells. For the latter algorithm we modified IL-8 secretion from CF cells by treatment with wild-type CFTR, with CPX, or by exposure to bacteria. The results have supported the hypothesis, and have provided a basis for considering the common pharmacogenomic expression signature as a surrogate endpoint for CF drug discovery. Significantly, the nature of the hypothesis, as well as the algorithm developed for this study, can be easily applied to pharmacogenomic studies with other goals. PMID- 12083970 TI - Gaucher disease: pediatric concerns. AB - Gaucher disease, the most prevalent lysosomal storage disorder, is inherited as an autosomal recessive condition. The gold standard for diagnosis is decreased acid beta-glucosidase activity in the lymphocytes or fibroblasts; molecular analysis of mutations allows for some prognostication of disease severity. Prenatal diagnosis and carrier testing for at-risk families are currently available. There is tremendous phenotypic heterogeneity in the non-neuronopathic form (type I), ranging from clinically asymptomatic to massive hepatomegaly, hypersplenism, growth retardation in children and extensive involvement of bone and lungs. Presence on one allele of the most common mutation, N370S, which is the most prevalent among Ashkenazi Jews for whom there is a predilection for Gaucher disease, is protective of neurological involvement. Some mutations, such as 84GG and IVS2+1, are associated with more severe disease manifestations when appearing as compound heterozygotes with N370S, but when occurring in the homozygous state are not compatible with life. Other mutations, such as L444P, are associated with severe non-neurological disease when occurring as compound heterozygotes with N370S, but when occurring in the homozygous state may be predictive of neurological disease of either acute (type II) or subacute (type III) forms. In the past decade, enzyme replacement therapy has become available which has resulted in a reduction in liver and spleen volume and consequently improved anemia and thrombocytopenia in most patients. It has also engendered catch-up growth in many children, induced improvement in lung involvement secondary to Gaucher disease, and to some extent ameliorated episodes of bone pain. By virtue of treatment, many children who may have been severely affected no longer need to undergo splenectomy to treat hypersplenism, and therefore they are not at risk of bone involvement consequent to the loss of the preferred reservoir for lipid-laden 'Gaucher cells'. However, enzyme treatment is ineffective in reversing neurological signs, requires a lifelong commitment to intravenous infusions, thereby reducing quality of life, and is relatively expensive for many national health schemes. Hence, alternative forms of treatment, such as substrate balance, are being explored. Symptomatic management, including orthopedic surgery, pain relief for bone pain and even splenectomy, still has importance for patients with Gaucher disease. In addition, there is the potential for bone marrow transplantation and, in the future, gene therapy to be curative, particularly for patients with the neuronopathic forms. PMID- 12083971 TI - Treatment strategies in children with hemophilia. AB - Hemophilia is an inherited bleeding disorder caused by quantitative or qualitative defects in the synthesis of factor VIII (FVIII) or factor IX (FIX). Clinically, it is divided into severe, moderate and mild disease depending on the levels of FVIII or FIX in the blood. The bleeding tendency is most pronounced and can start at a very young age in severe hemophilia, which is characterized by repeated hemorrhage into the joints and muscles. Without treatment, these episodes lead to severe arthropathy, and there is also a high risk of lethal cerebral hemorrhage. The treatment of bleeding symptoms requires the correction of the coagulation defect. Factor concentrates have been available for 30 years, initially with the development of cryoprecipitate, subsequently with increasingly purified plasma-derived forms, and ultimately with recombinant clotting factor concentrates. The advantage of this highly effective therapy has been subdued by the outbreak of HIV and Hepatitis C infections in patients with hemophilia treated with factor concentrates which did not have adequate viral inactivation steps in the purification process. Plasma-derived and recombinant factor concentrates are today considered to have a good safety profile, but are only available for a small group of hemophilia patients worldwide. A multidisciplinary team approach is important for early diagnosis, communication with the patient and parents, and to tailor the best treatment possible with the amount of clotting factor concentrates available. The main goal of hemophilia treatment is to prevent bleeding symptoms and allow normal integration in social life. In patients with severe hemophilia, this can best be achieved by early home treatment and primary prophylaxis. Future developments in gene therapy may transform severe hemophilia to a mild form, with no need for regular injections of clotting factor concentrates. PMID- 12083972 TI - Controlled hypotension in children: a critical review of available agents. AB - Due to the potential for the transmission of infectious diseases with the homologous transfusion of blood products, there has been an increased interest in measures to limit intraoperative blood loss and avoid the need for homologous transfusion during high-risk surgical procedures including spinal surgery. Controlled hypotension (also referred to as deliberate or induced hypotension), defined as a reduction of systolic blood pressure to 80 to 90 mm Hg, a reduction of mean arterial pressure (MAP) to 50 to 65 mm Hg or a 30% reduction of baseline MAP, is one technique that has been used to limit intraoperative blood loss. In the adult population, several agents have been used alone or in combination for controlled hypotension including the inhalational anesthetic agents, direct acting vasodilators such as nitroglycerin (glyceryl trinitrate) and nitroprusside, beta-adrenoceptor antagonists, and calcium channel antagonists. Despite clinical studies that have clearly demonstrated a reduction in blood loss with controlled hypotension when compared with the normotensive state and despite potential theoretical issues with each agent, there are no definitive studies demonstrating the preferred pharmacologic agent. When considering the pediatric aged patient, studies have reported the use of the inhalational agent sevoflurane, the alpha(2)-adrenoceptor agonist dexmedetomidine as well as various vasodilators including sodium nitroprusside, nitroglycerin, fenoldopam, and alprostadil for controlled hypotension. Sevoflurane offers the advantages of easy dosage titration, no need for an additional intravenous infusion as well as providing anesthesia in addition to controlled hypotension. Disadvantages include a slightly higher cost than some of the intravenous agents and the inability to monitor evoked potentials with high sevoflurane concentrations. Whereas sodium nitroprusside, nicardipine and fenoldopam all provide the desired level of hypotension in pediatric-aged patients, nitroglycerin was not effective in this age group of patients in one study. When comparing nicardipine and sodium nitroprusside, nicardipine offers the potential advantages of fewer episodes of excessive hypotension, less rebound tachycardia and, in one study, less blood loss. Although fenoldopam has been shown to be effective, cost issues may limit is widespread application for this technique. The pharmacologic profile of dexmedetomidine indicates that this drug has potential in controlled hypotension and clinical data are needed to define its role. PMID- 12083973 TI - Nebulized antibiotics in cystic fibrosis. AB - Nebulization is a useful administration route in cystic fibrosis (CF) as it delivers antibiotics directly to the endobronchial site of infection and is associated with decreased toxicity because of limited systemic absorption. It is assumed that the concentration of antibiotics in bronchial secretions should be as high as 10 times the minimum inhibiting concentration to allow penetration of antibiotics into biofilms, suppress inhibitory factors and promote bactericidal effectiveness. However, effective aerosol delivery is compromised by nebulizers with limited capacity to produce particles of a size in the respirable range. Three antibiotics are commonly used for inhalation: tobramycin, amikacin and colistin (colomycin). Placebo-controlled studies evaluating antibiotic aerosol maintenance in stable patients chronically infected with Pseudomonas aeruginosa indicate a significant improvement of lung function and a reduction of the number of hospital admissions for an acute exacerbation of CF. TOBI is a recently marketed preservative- and sulfate-free formula of tobramycin, specially designed for diffusion in the bronchioles and optimal tolerance. A wide-scope study involving 520 patients compared TOBI (300 mg twice daily; n = 258) with placebo (n = 262) for three 28-day cycles with each cycle separated by a 28-day period of no treatment. Respiratory function was significantly improved as early as in the second week and remained so for the rest of the trial even during periods without aerosol treatment. There was also a parallel decrease in the relative risk of hospitalization, the number of days of hospitalization and the number of days on intravenous antipyocyanic treatment. Toxicity studies carried out so far have shown no renal or ototoxicity with nebulized tobramycin. Introduction or selection of resistant bacteria is relatively rare but remains a matter of concern. Aerosol maintenance treatment with an appropriate antibiotic in a high enough dosage can be recommended for patients with CF who are chronically infected with P. aeruginosa. PMID- 12083974 TI - Efficacy and tolerability of extended-interval aminoglycoside administration in pediatric patients. AB - Aminoglycosides are commonly used to treat serious Gram-negative infections in pediatric patients. An effort to improve the efficacy and tolerability of this antibiotic class has led to evaluation of extended-interval aminoglycoside administration (EIAA). EIAA is designed to achieve higher peak plasma aminoglycoside concentrations, with relatively undetectable trough concentrations, when compared with conventional aminoglycoside administration (CAA), and is therefore expected to be markedly effective and to reduce drug accumulation and prevent nephrotoxicity and ototoxicity. Clinical trials evaluating EIAA in neonates included patients with suspected Gram-negative infections requiring short courses of aminoglycoside therapy. Consequently, comparative efficacy of EIAA versus CAA could not be assessed. In addition, ototoxicity was often not assessed, and nephrotoxicity was virtually undetectable. Similarly, trials evaluating EIAA versus CAA in infants and children have not demonstrated a difference in outcomes. The use of EIAA in children with febrile neutropenia has been evaluated primarily with amikacin. The incidences of nephrotoxicity and ototoxicity were low, and were similar between EIAA and CAA. No deaths were reported in any of these studies; however, this could be related to the inclusion of patients with undocumented bacteremia. Further investigation of EIAA is necessary in patients with documented bacteremia, since plasma aminoglycoside concentrations were undetectable for most of the dosage interval in children with febrile neutropenia who were treated once daily. Overall, clinical studies suggest that EIAA has similar efficacy to, and no higher risk of toxicity than, CAA in neonates, infants, and children. A few evaluations have also demonstrated that EIAA is cost-effective in neonates and in children with febrile neutropenia. Future studies evaluating the efficacy and tolerability of EIAA in pediatric patients with documented systemic infections should be prospective, randomized, controlled trials with sample sizes sufficient to detect differences between administration methods. Further evaluations should also address the optimal dosage and cost-effectiveness of EIAA in infants and children. PMID- 12083976 TI - Clinical pharmacokinetics and pharmacodynamics of repaglinide. AB - Repaglinide is a novel, fast-acting prandial oral hypoglycaemic agent developed for the treatment of patients with type 2 diabetes whose disease cannot be controlled by diet and exercise alone. Although repaglinide binds to the sulphonylurea binding sites on pancreatic beta-cells and has a similar mechanism of action, repaglinide exhibits distinct pharmacological properties compared with these agents. Following administration, repaglinide is absorbed rapidly and has a fast onset of dose-dependent blood-glucose lowering effect. The drug is eliminated rapidly via the biliary route, without accumulation in the plasma after multiple doses. Repaglinide is well tolerated in patients with type 2 diabetes, including elderly patients and patients with hepatic or renal impairment. The pharmacokinetic profile of repaglinide and the improvements in post-prandial hyperglycaemia and overall glycaemic control make repaglinide suitable for administration preprandially, with the opportunity for flexible meal arrangements, including skipped meals, without the risk of hypoglycaemia. PMID- 12083975 TI - Cytochrome p450 phenotyping/genotyping in patients receiving antipsychotics: useful aid to prescribing? AB - Many antipsychotics, including perphenazine, zuclopenthixol, thioridazine, haloperidol and risperidone, are metabolised to a significant extent by the polymorphic cytochrome P450 (CYP) 2D6, which shows large interindividual variation in activity. Significant relationships between CYP2D6 genotype and steady-state concentrations have been reported for perphenazine, zuclopenthixol, risperidone and haloperidol when used in monotherapy. Other CYPs, especially CYP1A2 and CYP3A4, also contribute to the interindividual variability in the kinetics of antipsychotics and the occurrence of drug interactions. For many antipsychotics, the role of the different CYPs at therapeutic drug concentrations remains to be clarified. Some studies have suggested that poor metabolisers for CYP2D6 would be more prone to oversedation and possibly parkinsonism during treatment with classical antipsychotics, whereas other, mostly retrospective, studies have been negative or inconclusive. For the newer antipsychotics, such data are lacking. Whether phenotyping or genotyping for CYP2D6 or other CYPs can be used to predict an optimal dose range has not been studied so far. Genotyping or phenotyping can today be recommended as a complement to plasma concentration determination when aberrant metabolic capacity (poor or ultrarapid) of CYP2D6 substrates is suspected. The current rapid developments in molecular genetic methodology and pharmacogenetic knowledge can in the near future be expected to provide new tools for prediction of the activity of the various drug-metabolising enzymes. Further prospective clinical studies in well-defined patient populations and with adequate evaluation of therapeutic and adverse effects are required to establish the potential of pharmacogenetic testing in clinical psychiatry. PMID- 12083977 TI - Clinical pharmacokinetics of depot leuprorelin. AB - Leuprorelin acetate is a synthetic agonist analogue of gonadotropin-releasing hormone. Continued leuprorelin administration results in suppression of gonadal steroid synthesis, resulting in pharmacological castration. Since leuprorelin is a peptide, it is orally inactive and generally given subcutaneously or intramuscularly. Sustained release parenteral depot formulations, in which the hydrophilic leuprorelin is entrapped in biodegradable highly lipophilic synthetic polymer microspheres, have been developed to avoid daily injections. The peptide drug is released from these depot formulations at a functionally constant daily rate for 1, 3 or 4 months, depending on the polymer type [polylactic/glycolic acid (PLGA) for a 1-month depot and polylactic acid (PLA) for depot of >2 months], with doses ranging between 3.75 and 30mg. Mean peak plasma leuprorelin concentrations (C(max)) of 13.1, 20.8 to 21.8, 47.4, 54.5 and 53 microg/L occur within 1 to 3 hours of depot subcutaneous administration of 3.75, 7.5, 11.25, 15 and 30 mg, respectively, compared with 32 to 35 microg/L at 36 to 60 min after a subcutaneous injection of 1mg of a non-depot formulation. Sustained drug release from the PLGA microspheres maintains plasma concentrations between 0.4 and 1.4 microg/L over 28 days after single 3.75, 7.5 or 15mg depot injections. Mean areas under the concentration-time curve (AUCs) are similar for subcutaneous or intravenous injection of short-acting leuprorelin 1mg; a significant dose-related increase in the AUC from 0 to 35 days is noted after depot injection of leuprorelin 3.75, 7.5 and 15mg. Mean volume of distribution of leuprorelin is 37L after a single subcutaneous injection of 1mg, and 36, 33 and 27L after depot administration of 3.75, 7.5 and 15mg, respectively. Total body clearance is 9.1 L/h and elimination half-life 3.6 hours after a subcutaneous 1mg injection; corresponding values after intravenous injection are 8.3 L/h and 2.9 hours. A 3 month depot PLA formulation of leuprorelin acetate 11.25mg ensures a C(max) of around 20 microg/L at 3 hours after subcutaneous injection, and continuous drug concentrations of 0.43 to 0.19 microg/L from day 7 until before the next injection. Recently, an implant that delivers leuprorelin for 1 year has been evaluated. Serum leuprorelin concentrations remained at a steady mean of 0.93 microg/L until week 52, suggesting zero-order drug release from the implant. In general, regular or depot leuprorelin treatment is well tolerated. Local reactions are more common after application of the 3- or 4-month depot in comparison with the 1-month depot. PMID- 12083978 TI - Beta-adrenergic blockers in systemic hypertension: pharmacokinetic considerations related to the current guidelines. AB - Beta-adrenergic blockade has provided one of the major pharmacotherapeutic advances of the 20th century. Beta-blockers are first-line drugs for the management of systemic hypertension, used alone and in combination with other antihypertensive agents. Drugs in the beta-blocking class have the common property of blocking the binding of catecholamines to beta-adrenergic receptor sites; however, there are significant pharmacodynamic and pharmacokinetic differences between the individual agents that are of clinical importance. Among these differences are the completeness of gastrointestinal absorption, the degree of hepatic first-pass metabolism, lipid solubility, protein binding, brain penetration, concentration within the cardiac tissue, rate of hepatic biotransformation, and renal clearance of drug and/or metabolites. Long-acting formulations of existing beta-blockers are currently in use, and ultra-short acting agents are also available. Age, race, cigarette smoking and concomitant drug therapy can also influence the pharmacokinetics of beta-blocking drugs. The wide interpatient variability in plasma drug concentrations observed with beta blockers makes this parameter unreliable in routine patient management. Despite the pharmacokinetic differences among beta-blockers, these drugs should always be titrated to achieve the desired individual patient response. PMID- 12083979 TI - Sevelamer, a phosphate-binding polymer, is a non-absorbed compound. AB - OBJECTIVE: To examine the absorption, distribution and excretion of sevelamer hydrochloride in rats and humans. PARTICIPANTS: Twelve male Sprague-Dawley rats were used in the animal study, and twenty human volunteers participated in the clinical trial. METHODS: In the animal study, six rats received a single oral dose of [(3)H]sevelamer and six rats were pretreated with unlabelled sevelamer in the diet for 28 days followed by a single dose of [(3)H]sevelamer on day 29. Total urine and faeces were collected at intervals up to 72 hours post dose, and tissues were obtained at the time of sacrifice. In the clinical trial, subjects received a single oral dose of [(14)C]sevelamer following 28 days of pretreatment with unlabelled sevelamer. Blood, urine and faeces samples were collected at intervals up to 96 hours. RESULTS: In the rat study, no significant urinary excretion of radioactivity was observed. The average recovery of radioactivity in the faeces was 98% in the single-dose group and greater than 100% in the group pretreated with unlabelled sevelamer for 28 days. A total of less than 0.1% of the dose was found in the tissues. In the human study, no detectable amount of (14)C was found in the blood of any subject at any time. The majority of subjects had no detectable amounts of (14)C recovered in the urine. In subjects where (14)C was recovered in the urine, less than 0.02% was detected, a level equivalent to the free (14)C detected in the [(14)C]sevelamer preparation. On average, greater than 99% of the administered dose was recovered in the faeces of the subjects. CONCLUSION: These studies demonstrate that sevelamer is a non absorbed compound. PMID- 12083981 TI - Could statins prevent age-related macular degeneration? PMID- 12083980 TI - Pharmacokinetics of cetirizine in tear fluid after a single oral dose. AB - BACKGROUND: Antihistamines (histamine H(1) receptor antagonists) are effective and convenient drugs for the treatment of allergic conjunctivitis. Because of the short duration of action generally observed for drugs administered topically to the eye, the oral route is often preferred. However, the presence of a selective barrier between blood and ocular tissues, the so-called blood-ocular barrier, does not allow a priori assessment of the most suitable dosage for ocular therapy. OBJECTIVE: To investigate the tear concentrations of cetirizine, a second-generation antihistamine, over time following oral administration, and to study the relationship between plasma and tear fluid concentrations. DESIGN AND PARTICIPANTS: Pharmacokinetic study of a single oral dose of cetirizine 10mg in 40 patients treated for allergic conjunctivitis. METHODS: Patients received a single oral dose of cetirizine. Samples of blood and tear fluid were taken according to predefined sampling schedules and the concentrations of cetirizine were determined by a new high performance liquid chromatography method. RESULTS: Concentration-time profiles for cetirizine in serum and tear fluid were similar, although the mean maximum concentration in tear fluid was reached later than in serum (90 and 30 min, respectively). However, at 60 and 120 min the cetirizine concentration in tear fluid was 98 and 92% of the mean maximum concentration, respectively, showing a plateau region and indicating that the disposition rate for the tear fluid compartment was very similar to that for the blood compartment. CONCLUSION: Oral administration of cetirizine yields therapeutically effective concentrations of the drug at the anterior surface of the eye. PMID- 12083982 TI - Therapy of Crohn's disease in childhood. AB - Crohn's disease in childhood is a chronic relapsing and remitting condition that can significantly impact on normal growth and development. This influences the choice of both initial and ongoing management. The goal of therapy is to induce and maintain remission with minimal side effects. Enteral nutrition as the sole therapy for active disease is effective in some children, thus avoiding the use of corticosteroids. In disease that is resistant to conventional treatment, immunosuppression or anti-tumour necrosis factor therapy is indicated. We review the use of these treatments and discuss the new therapies being developed, including antibodies, cytokines and probiotics. PMID- 12083983 TI - Current pharmacotherapeutic strategies for overactive bladder. AB - Overactive bladder (OAB) is a chronic, distressing condition characterised by symptoms of urgency (sudden overwhelming urge to urinate) and frequency (urinating more than eight times daily), with or without urge urinary incontinence (sudden involuntary loss of urine). It affects millions of people of all ages and both sexes world wide, with greater prevalence in women and the elderly. The treatment of OAB is aimed at reducing debilitating symptoms, which have a significant effect on all aspects of an individual's quality of life, including social, domestic, psychological, occupational, physical and sexual functioning. Anticholinergic agents are currently recommended as first-line therapy for OAB. Their use results in significant clinical improvement in patients, although a lack of selectivity for receptors in the bladder may lead to troublesome side effects, including dry mouth, blurred vision, somnolence, dizziness and constipation. Recent research efforts have focused on developing drugs with a reduced propensity for causing these problems. Of the available anticholinergic agents, oxybutynin and tolterodine are the most widely used to treat OAB. Studies directly comparing tolterodine immediate-release (IR) with oxybutynin IR have shown that the two agents have similar efficacy. However, tolterodine IR is significantly better tolerated, particularly with respect to the incidence and severity of dry mouth. An extended-release formulation of tolterodine (4 mg capsules) has recently been developed to allow for once-daily dosing. In addition to greater convenience, tolterodine extended-release has shown enhanced efficacy and tolerability compared with tolterodine IR. PMID- 12083984 TI - Current and emerging therapies in osteoporosis. AB - Osteoporosis is a systemic skeletal disorder leading to decreased bone mineralisation and a propensity for fracture. The disease affects millions of people and is a source of significant morbidity and mortality. Fracture-related costs have skyrocketed in recent years and projections suggest an increase as the population ages. Recent advances in the understanding of bone biology have improved the therapeutic options for osteoporosis. This is a review of the osteoporotic therapies that have recently become available and a discussion of emerging options. An overview on current practice guidelines is provided. PMID- 12083985 TI - Advances in the treatment of rheumatoid arthritis: old versus new therapies. AB - Rheumatoid arthritis (RA) is a common cause of disability in the western population, with an annual incidence of 0.05% and a prevalence of 1%. Although a small percentage of patients go into natural remission, the untreated disease progresses to cause disability, morbidity and early mortality. Unravelling of the cytokine network in the pathogenesis of RA has led to the development of drugs that target these cytokines and prevent joint damage. Three biological anticytokine agents, etanercept, infliximab and anakinra, are now available for use in RA. More experience will quantify their safety and benefits. The potency of the older disease-modifying antirheumatic drugs (DMARDs), such as methotrexate and sulfasalazine, is also being realised, especially when used early in the disease process and in combination. Leflunomide is a new DMARD with efficacy similar to methotrexate and sulfasalazine. Symptomatic treatment of RA with nonsteroidal anti-inflammatory drugs has also undergone a revolution with the availability of a new class of COX-2-specific inhibitors. These drugs control inflammation and provide pain relief with less GI toxicity. Management of comorbid conditions associated with RA and its treatment (i.e., osteoporosis, cardiovascular and lung disease) has also become a priority for the rheumatologist. It is hoped that more aggressive use of conventional DMARDs and biological agents will result in less disability and a higher proportion of patients achieving remission. PMID- 12083986 TI - Current treatment options for chronic granulomatous disease. AB - Chronic granulomatous disease is a rare inherited disorder of phagocytic cells which results in a susceptibility to infections of catalase-positive bacteria and fungi (especially Aspergillus species), as well as granuloma formation. The mainstay of therapy is antibacterial and antifungal prophylaxis. Trimethoprim sulfamethoxazole is the drug of choice for the prevention of bacterial infection, while itraconazole is most widely used for the prevention of fungal infection. Immunomodulatory agents, such as IFN-phi, have a role in the prevention and treatment of intractable infection. New antifungal agents provide the promise of improved cure rates for invasive Aspergillus, while bone marrow transplants and gene therapy may offer the promise of complete cure. PMID- 12083987 TI - Hormonal therapies in acne. AB - Hormones, in particular androgen hormones, are the main cause of acne in men, women, children and adults, in both normal states and endocrine disorders. Therefore, the use of hormonal therapies in acne is rational in concept and gratifying in practice. Although non-hormonal therapies enjoy wide usage and continue to be developed, there is a solid place for hormonal approaches in women with acne, especially adult women with persistent acne. This review covers the physiological basis for hormonal influence in acne, the treatments that are in use today and those that show promise for the future. The main treatments to be discussed are oral contraceptives androgen receptor blockers like spironolactone and flutamide, inhibitors of the enzyme 5 alpha-reductase and topical hormonal treatments. PMID- 12083988 TI - Pharmacotherapy of acute respiratory distress syndrome. AB - To date, the only therapeutic option that has convincingly been shown to decrease mortality in acute respiratory distress syndrome (ARDS) has been to use a lung protective strategy that minimises the iatrogenic consequences of providing adequate life support through the use of mechanical ventilation. In terms of the pharmacological options for ARDS, no single drug or treatment has been shown to be the magic bullet in this disease. The search for novel therapies and pharmacological agents is active and relentless. Important pathophysiological areas of focus are preventative therapy, supportive care and treatment of the underlying inflammatory process. In this paper we will review current and experimental approaches to the management of ARDS. In addition, the pathophysiological basis for their putative modes of action, the current state of the literature and the potential for future clinical development will be discussed. PMID- 12083989 TI - Oxygen free radicals and the penis. AB - Penile erection is dependent upon vascular smooth muscle relaxation in erectile tissue and penile arteries, the principal mediator of relaxation being nitric oxide (NO). Evidence from basic scientific studies indicates that oxidative stress mediated through the superoxide radical (superoxide) and other reactive oxygen species (ROS) may be central to impaired cavernosal function in erectile dysfunction (ED). Increased inactivation of NO by superoxide results in impaired penile NO transmission and smooth muscle relaxation. Furthermore, propagation of endothelial dysfunction by ROS may result in chronic impairment of penile vascular function, a process analogous to early atherogenesis. Indeed, ED and atherosclerosis are closely linked through shared risk factors. Given our current understanding of ED pathophysiology, antioxidants may be of benefit in both the short- and long-term. Evidence supporting the paradigm of antioxidant therapy for the prevention or treatment of ED is presented herein. PMID- 12083990 TI - The treatment of Tourette's syndrome: current opinions. AB - Correct education of the patient is one of the most important aspects in the treatment of Tourette's syndrome. Pharmacotherapy is often unsatisfactory and therefore should be limited to those patients who are significantly impaired. Therapy must be individualised and the most troublesome symptom should be targeted first. In the treatment of tics, dopamine receptor blocking agents are currently the most effective drugs. It is currently unknown whether classic neuroleptics, such as pimozide, selective dopamine receptor antagonists, such as sulpiride, or newer atypical antipsychotics, such as risperidone, have the best adverse effect profile. Tiapride can be used as an alternative, particularly in children. Selective serotonin-re-uptake inhibitors are recommended for the treatment of obsessive-compulsive behaviour. In children suffering from attention deficit hyperactivity disorder, psychostimulants, such as methylphenidate, are the treatment of choice. Recent studies have provided increasing evidence that stimulants do not cause a significant increase in tics in the majority of patients. PMID- 12083991 TI - Present and future pharmacotherapy for heart failure. AB - The pharmacotherapy currently recommended by the American College of Cardiology and the American Heart Association for heart failure (HF) is a diuretic, an angiotensin-converting enzyme inhibitor (ACEI), a beta-adrenoceptor antagonist and (usually) digitalis. This current treatment of HF may be improved by optimising the dose of ACEI used, as increasing the dose of lisinopril increases its benefits in HF. Selective angiotensin receptor-1 (AT(1)) antagonists are effective alternatives for those who cannot tolerate ACEIs. AT(1) antagonists may also be used in combination with ACEIs, as some studies have shown cumulative benefits for the combination. In addition to being used in Stage IV HF patients, in whom it has a marked benefit, spironolactone should be studied in less severe HF and in the presence of beta-blockers. The use of carvedilol, extended-release metoprolol and bisoprolol should be extended to severe HF patients as these agents have been shown to decrease mortality in this group. The ancillary properties of carvedilol, particularly antagonism at prejunctional beta adrenoceptors, may give it additional benefits to selective beta(1)-adrenoceptor antagonists. Celiprolol and bucindolol are not the beta-blockers of choice in HF, as they do not decrease mortality. Although digitalis does not reduce mortality, it remains the only option for a long-term positive inotropic effect, as the long term use of the phosphodiesterase inhibitors is associated with increased mortality. The calcium sensitising drug levosimendan may be useful in the hospital treatment of decompensated HF to increase cardiac output and improve dyspnoea and fatigue. The antiarrhythmic drug amiodarone should probably be used in patients at high risk of arrhythmic or sudden death, although this treatment may soon be superseded by the more expensive implanted cardioverter defibrillators, which are probably more effective and have fewer side effects. The natriuretic peptide nesiritide has recently been introduced for the hospital treatment of decompensated HF. Novel drugs that may be beneficial in the treatment of HF include the vasopeptidase inhibitors and the selective endothelin A receptor antagonists but these require much more investigation. However, disappointing results have been obtained in a large clinical trial of the tumour necrosis factor alpha antagonist etanercept, where no likelihood of a difference between placebo and etanercept was observed. Small clinical trials with recombinant growth hormone to thicken ventricles in dilated cardiomyopathy have given variable results. PMID- 12083992 TI - An update on photodynamic therapy in age-related macular degeneration. AB - Age-related macular degeneration (AMD) is the leading cause of irreversible loss of central vision in people aged > 50 years in the western world. Until recently, the only proven treatment to reduce the risk of vision loss from its more severe neovascular form was laser photocoagulation, but this treatment was suitable for only 15% of cases. Photodynamic therapy (PDT) with verteporfin was recently proposed to be effective in reducing the risk of visual loss for an estimated 20 30% of neovascular AMD patients. This review covers AMD epidemiology, the mechanism of PDT, the 2-year results of the two major clinical studies of PDT with verteporfin, the cost-effectiveness of PDT and the current research status of other drugs for PDT in AMD. PMID- 12083993 TI - Recurrent and/or metastatic thyroid cancer: therapeutic options. AB - Thyroid cancer is relatively rare, accounting for 0.5 - 10 cases per 100,000 individuals per year. Despite their generally favourable prognosis, patients with differentiated thyroid cancer are at risk of tumour recurrence for decades after diagnosis. The optimal management remains controversial even in the low-risk patients because of the high cure rates, long natural history and rarity of these tumours. Therapeutic interventions in recurrent and metastatic differentiated thyroid cancer depend on the type of initial treatment, the site and the extent of disease. Surgical excision of the amenable-to-surgery lesions and radioiodine administration remain the first approach. External radiotherapy may be given to patients with inoperable lesions or those not concentrating radioiodine. Chemotherapy has not provided consistently successful results. Various therapeutic approaches for anaplastic carcinoma give poor results, making the development of novel treatments necessary. Innovative strategies, including recombinant human thyroid stimulating hormone, retinoic acid redifferentiation therapy and gene therapy, may lead to further improvement in the management of thyroid cancer arising from follicular cells. PMID- 12083994 TI - Allopathic and complementary alternatives to hormone replacement therapy. AB - Management of the menopause is a rapidly growing concern due to the ageing human population. The overall female lifespan has increased over the last century and up to a third of a woman's life is now spent in menopause. To that end, significant attention has been placed on maximising the quantity and quality of life in the menopausal years. The optimal management strategies are ones that are highly flexible and sensitive to an individual's expectations and concerns. Thus, while traditional oestrogen replacement therapy has been in place for > 20 years, there is now a greater interest in alternatives to this modality for those women who cannot or will not use it. This article reviews some of the alternative therapies that are being incorporated both in the allopathic and complementary medicine arenas. PMID- 12083995 TI - Entacapone in the management of Parkinson's disease. AB - Entacapone is one of a new class of drugs, the catechol-O-methyltransferase (COMT) inhibitors, which expand the therapeutic options for Parkinson's disease by extending the action of levodopa. Entacapone is used in conjunction with levodopa and provides benefit to patients who suffer from motor fluctuations. Side effects include worsening of the dyskinesias associated with peak doses of levodopa, hypotension, constipation and urine discoloration. Unlike tolcapone, an earlier COMT inhibitor, entacapone does not require liver function monitoring. PMID- 12083996 TI - Travoprost--a new prostaglandin analogue for the treatment of glaucoma. AB - Travoprost, a highly selective and potent analogue of the prostaglandin PGF(2)(alpha), has recently been approved and marketed as a topical ocular hypotensive agent for the treatment of ocular hypertension and glaucoma. Following absorption into the eye, the free acid form of travoprost interacts with the endogenous FP prostanoid receptor to enhance aqueous humor outflow and lower intraocular pressure (IOP). Travoprost is distinguished from other marketed prostaglandin analogues in that it is a full agonist at the prostaglandin receptor. It is also highly selective with little or no affinity for other prostanoid or non-prostanoid receptors in the eye. Travoprost provides robust lowering of IOP with little diurnal fluctuation and results in low target pressures in a large percentage of patients. In controlled clinical trials, travoprost 0.004% o.d. used as monotherapy produced greater IOP reduction than timolol 0.5% b.i.d. and equal or greater reduction than latanoprost 0.005%o.d. Travoprost 0.004% was also shown to be an effective adjunctive agent offering an additional 5 - 7 mmHg IOP reduction in patients inadequately controlled on timolol 0.5%. Subgroup analysis of a large Phase III trial revealed travoprost 0.004% to be significantly more effective at lowering IOP in African American patients by almost 2 mmHg compared to non-African Americans. Moreover, a higher percentage of African American patients responded to travoprost 0.004% and reached lower target pressures than with either latanoprost 0.005% or timolol 0.5%. Travoprost is a very stable compound, maintaining its efficacy following exposure to extremely low and high temperatures, repeated freezing and thawing and exposure to light. Throughout all clinical trials, travoprost was found to be safe and well-tolerated with very few (< 5%) discontinuations due to adverse events. Travoprost 0.004% represents a clinically significant advance for the treatment of glaucoma and ocular hypertension, offering superior IOP reduction and diurnal control, especially among African American patients, in a safe, well tolerated, stable formulation. PMID- 12083997 TI - Review of fluoxetine and its clinical applications in premenstrual dysphoric disorder. AB - The largest number of antidepressant treatment trials in premenstrual syndrome and premenstrual dysphoric disorder (PMDD) have been conducted with fluoxetine. Fluoxetine and other selective serotonin re-uptake inhibitors (SSRIs) clearly reduce premenstrual emotional and physical symptoms and improve premenstrual psychosocial functioning. Fluoxetine was the first SSRI to be approved by the FDA as a treatment for the emotional and physical symptoms of PMDD. Fluoxetine 20 mg has been reported to be effective for emotional and physical premenstrual symptoms with continuous daily dosing (every day of the menstrual cycle) and with luteal phase daily dosing (from ovulation to menses). In addition, premenstrual emotional symptoms have been reported to improve with fluoxetine 10 mg in luteal phase daily dosing and with 90 mg 2 and 1 weeks prior to menses. Fluoxetine is generally a well-tolerated treatment for PMDD and discontinuation effects have not been reported with intermittent dosing regimens. PMID- 12083998 TI - Review of zolmitriptan and its clinical applications in migraine. AB - Preclinical studies have shown that zolmitriptan is a selective serotonin 5 HT(1B/1D) receptor agonist (triptan). Randomised, placebo-controlled, double blind trials in patients with migraine have shown that zolmitriptan has good efficacy measured using 2 h response and pain-free rates. Migraine-associated symptoms, including nausea, photophobia and phonophobia, are also improved with zolmitriptan. Oral zolmitriptan (2.5 and 5 mg) has an onset of action within 45 min and efficacy is sustained in most patients who respond at 2 h. The orally disintegrating zolmitriptan tablet has the advantage that it may be taken immediately, without the need for additional fluids, any time a migraine headache occurs. Patients may benefit in terms of improved efficacy from the convenience of the disintegrating tablet, since there is evidence that taking triptan therapy as early as possible in an attack is advantageous. For similar reasons, as well as improved efficacy, a nasal spray formulation is in development. Zolmitriptan is effective in the treatment of migraine associated with menses and migraine with aura. There is no tachyphylaxis following repeated doses for multiple attacks of migraine over a prolonged period of time. Compared to placebo, the incidence of persistent migraine headache is reduced by zolmitriptan and recurrent migraine headache occurs less frequently. Zolmitriptan has also shown efficacy in the treatment of persistent and/or recurrent migraine headache. Comparative clinical studies have shown overall that zolmitriptan has similar or superior efficacy to sumatriptan in the treatment of migraine. Specifically, zolmitriptan 2.5 mg was significantly more effective than sumatriptan 25 or 50 mg according to a number of end points, including headache response at 2 h. Oral zolmitriptan is also effective in the acute treatment of cluster headache. Zolmitriptan is generally well tolerated, with most adverse events being mild-to moderate, transient and resolving without intervention or the need for treatment withdrawal. The consistent efficacy in treating all types of migraine and the choice of available formulations make zolmitriptan acceptable to patients and a suitable first-line therapy for the treatment of migraine. PMID- 12083999 TI - Zoledronate once-yearly increases bone mineral density--implications for osteoporosis. AB - Osteoporosis is a major and growing problem for older women and men in western society. The bisphosphonates are used to treat postmenopausal osteoporosis because they decrease the risk of fractures. Oral bisphosphonates may cause oesophageal irritation and injury and are poorly absorbed. Intermittent intravenous bisphosphonates, including zoledronate, may be a way to avoid the problems associated with oral administration. Intravenous zoledronate increased bone mineral density while decreasing bone turnover and formation. Interestingly, the effects of a single dose (4 mg) of zoledronate were maintained over a year and were similar to those of three-monthly dosing with 0.25 - 1 mg zoledronate. If the increases in bone mineral density with zoledronate translate into reduced fractures, once-yearly intravenous zoledronate could be simple prophylaxis for postmenopausal osteoporosis. PMID- 12084000 TI - Metformin & lifestyle intervention prevent Type 2 diabetes: lifestyle intervention has the greater effect. AB - Diabetes mellitus is now occurring in epidemic proportions in many countries. Owing to the limited effectiveness of drug prophylaxis of diabetic complications after diabetes has developed, it may be more appropriate to investigate ways to prevent the onset of diabetes. A recent trial published by the Diabetes Prevention Programme Research Group investigated whether lifestyle changes or metformin were effective in delaying the onset of diabetes in subjects with impaired glucose tolerance. The goals of the intensive lifestyle intervention were to achieve and maintain a weight reduction of 7% through a low-calorie, low fat diet and to engage in physical activity of moderate intensity, such as brisk walking, for at least 150 min/week. The effectiveness of the intensive lifestyle intervention on body weight was initially quite good but decreased over time. Metformin caused some weight loss but to a lesser extent than the intensive lifestyle intervention. Both therapies decreased the fasting plasma glucose levels to a similar extent initially. The intensive lifestyle intervention decreased plasma glycosylated haemoglobin levels to a greater extent than metformin. Both intensive lifestyle intervention and metformin reduced the incidence of diabetes, with the lifestyle intervention having the greater effect. PMID- 12084002 TI - Antibiotics in the treatment of acute exacerbations of chronic bronchitis. AB - The benefit of antimicrobial therapy for patients with an acute exacerbation of chronic bronchitis (AECB) remains controversial for two main reasons. First, the distal airways of patients with chronic bronchitis are persistently colonised, even during clinically stable periods, with the same bacteria that have been associated with AECB. Second, bacterial infection is only one of several causes of AECB. These factors have led to conflicting analyses on the role of bacterial agents and the response to antimicrobial therapy of patients with AECB. An episode of AECB is said to be present when a patient with chronic obstructive pulmonary disease (COPD) experiences some combination of increased dyspnoea, increased sputum volume, increased sputum purulence and worsening lung function. While the average COPD patient experiences 2 - 4 episodes of AECB per year, some patients, particularly those with more severe airway obstruction, are more susceptible to these attacks than others. Bacterial agents appear to be particularly associated with AECB in patients with low lung function and those with frequent episodes accompanied by purulent sputum. Non-typeable Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis account for up to 50% of episodes of AECB. Gram-negative bacilli are more likely to occur in patients with more severe lung disease. Antibiotics have been used to ameliorate AECB, to prevent AECB and to prevent the long-term loss of lung function that characterises COPD. Numerous prevention trials have been conducted with fairly consistent results; antibiotics do not lessen the number of episodes of AECB but do reduce the number of days lost from work. Most antibiotic trials have studied the impact of treatment on episodes of AECB and results have been inconsistent, largely due to patient selection and end point definition. In patients with severe airway obstruction, especially in the presence of purulent sputum, antibiotic therapy significantly shortens the duration of symptoms and can be cost-effective. Over the past 50 years, virtually all classes of antimicrobial agents have been studied in AECB. Important considerations include penetration into respiratory secretions, spectrum of activity and antimicrobial resistance. These factors limit the usefulness of drugs such as amoxicillin, erythromycin and trimethoprim-sulfamethoxazole. Extended-spectrum oral cephalosporins, newer macrolides and doxycycline have demonstrated efficacy in clinical trials. Amoxicillin-clavulanate and flouoroquinolones should generally be reserved for patients with more severe disease. A number of investigational agents, including ketolides and newer quinolones, hold promise for treatment of AECB. PMID- 12084001 TI - Inhaled diazeniumdiolates (NONOates) as selective pulmonary vasodilators. AB - Selective pulmonary vasodilators cause vasodilatation limited to the pulmonary vasculature, within well-ventilated lung regions. Selective pulmonary vasodilators ideally cause only a minimal effect on the systemic circulation and improve ventilation/perfusion matching. NONOates are a novel group of chemical compounds that spontaneously and continuously release nitric oxide under physiological conditions, over periods of up to 24 h. Inhaled NONOates retain the benefits of gaseous nitric oxide without many of its therapeutic disadvantages. This review focuses on the therapeutic potential of inhaled NONOates in pulmonary hypertension, other lung conditions associated with right ventricular dysfunction and in asthma. The potential toxicity of NONOates is also discussed. PMID- 12084003 TI - Rheumatoid arthritis: developing pharmacological therapies. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease that, despite recent advances in therapy, still results in significant morbidity, mortality and disability. The aetiology remains unknown and past therapies, although helpful for the majority of patients, have been suboptimal. The recent introduction of newer agents has changed the treatment paradigm of RA. COX-2 inhibitors, anti-TNF agents and interleukin-1 antagonists have allowed us to treat RA more effectively with relatively low risk of side effects. Investigations of other possible treatment pathways, such as inhibition of angiogenesis, may produce still better treatment and rapid unraveling of the immune system and how it relates to RA greatly enhances the opportunities for improved therapeutics in RA. PMID- 12084004 TI - Novel therapies in the treatment of spondyloarthritis. AB - Spondyloarthritis represents one of the commonest groups of inflammatory arthritides with onset in the third and fourth decades and primarily affecting the axial skeleton. Current treatment is primarily symptomatic, non-steroidal anti-inflammatory drugs being used most commonly. No therapeutics have been shown to prevent structural damage. The development of validated and standardised outcome instruments and a composite criterion of response should encourage evaluation of new therapeutics. Anti-TNF- alpha -directed therapeutics have been shown to be dramatically effective in short-term (12 week) placebo-controlled trials in both ankylosing spondylitis and psoriatic arthritis whilst observational cohorts describe efficacy that is maintained for over one year. Treatment has been well-tolerated, with mycobacterial infections being the primary concern. Significant costs and the requirement for continuous therapy are likely to spur the development of orally bioavailable agents targeting TNF- alpha expression. PMID- 12084005 TI - TNF inhibition as therapy for rheumatoid arthritis. AB - The introduction of TNF- alpha -inhibiting biologicals has been a major therapeutic breakthrough in rheumatoid arthritis therapy. Against a background of conventional disease-modifying antirheumatic drug experience, this review focuses on present experiences and possible future developments. TNF inhibition results in profound improvement in the majority of rheumatoid arthritis patients, but non response and adverse effects need attention. Adalimumab is being filed for approval. Other monoclonal antibodies or receptor constructs are in late development. Small molecule inhibitors of TNF production or signalling are a hot topic. One emerging target is nuclear factor kappa B and selective inhibition has proved effective in animal models of arthritis. Synovial proliferation in rheumatoid arthritis is characterised by diminished apoptosis of fibroblasts, whereas bone marrow precursor cells undergo accelerated apoptosis in active rheumatoid arthritis. Both abnormalities are seemingly ameliorated by TNF inhibition. Anti-apoptotic strategies will soon go into development for control of unresponsive rheumatoid arthritis. PMID- 12084006 TI - Cough: potential pharmacological developments. AB - Cough is an important defensive reflex of the upper airway and is also a very common symptom of respiratory disease. Cough following an upper respiratory viral infection is transient, and persistent cough is associated with a whole range of conditions, such as asthma, rhino-sinusitis and gastro-oesophageal reflux. Treatment directed at these conditions may improve the associated cough. There is often a need, however, to control cough itself whatever the cause. The most effective drugs in this class are the opioids, such as morphine, codeine or pholcodeine, but at effective doses they have side effects including drowsiness, nausea, constipation and physical dependence. Investigations into the cough reflex and into the potential mechanisms of sensitised cough reflex have uncovered several potential targets for novel drugs. New opioids apart from mu agonists such as kappa- and delta -receptor agonists, have been developed, in addition to non-opioids such as nociceptin. Neurokinin receptor antagonists, bradykinin receptor antagonists, vanniloid receptor VR-1 antagonists may be beneficial by blocking effects of tachykinins and sensory nerve activation. Local anaesthetics, blockers of sodium-dependent channels and maxi-K Ca2+-dependent channel activators of afferent nerves are inhibitors of the cough reflex. Some of these novel agents may act centrally or peripherally or at both sites as antitussives. Large scale trials of these novel compounds have not been carried out in cough in man but there is a serious need for more effective antitussives devoid of side effects. PMID- 12084007 TI - Neutrophil elastase inhibitors as treatment for COPD. AB - Chronic obstructive pulmonary disease, characterised by a slowly progressive, irreversible airways limitation, is a major worldwide cause of chronic morbidity and mortality. The imbalance between human neutrophil elastase and endogenous antiproteases may cause excess human neutrophil elastase in pulmonary tissues, which may be considered a major pathogenic factor in chronic obstructive pulmonary disease. Great effort has been devoted to finding a method to restore the balance, resulting in the discovery of potent two-typed small-molecular weight human neutrophil elastase inhibitors. In the application of chronic obstructive pulmonary disease therapy, the human neutrophil elastase inhibitors mainly focused upon include ONO-5046, MR-889, L-694,458, CE-1037, GW-311616 and TEI-8362 as the acyl-enzyme inhibitors; and ONO-6818, AE-3763, FK-706, ICI 200,880, ZD-0892 and ZD-8321 as the transition-state inhibitors. In this review, various problems that remain to be solved in the clinical use of human neutrophil elastase inhibitors are discussed. PMID- 12084008 TI - Advances in pathogenic concepts and therapeutic agents in Rasmussen's encephalitis. AB - Rasmussen's encephalitis is a rare inflammatory brain disease which occurs mainly in children and is characterised by affection of only one hemisphere. Pathogenetic concepts have considered three different, not mutually exclusive, key factors contributing to the initiating or perpetuating events in the central nervous system. These include viruses, autoimmune antibodies and autoimmune cytotoxic T lymphocytes. Based on these concepts, different therapeutic strategies have been pursued, such as antiviral agents, plasmapheresis, immuno adsorption, immunosuppression or immunomodulation with intravenous immunoglobulins. However, due to the lack of larger studies, to date there is no established therapeutic strategy of this devastating disease. An overview of the current state of immunepathogenic concepts for Rasmussen's encephalitis is given and past and present treatment attempts are discussed, including an outline of future perspectives. An opinion on symptomatic treatment with anticonvulsive drugs is included. PMID- 12084009 TI - Bosentan: a dual endothelin receptor antagonist. AB - The peptide endothelin plays a significant role in a wide array of pathological conditions, including primary pulmonary hypertension and pulmonary arterial hypertension associated with collagen vascular disease. These are life threatening conditions that can severely compromise the function of the lungs and heart. Inhibiting the actions of endothelin by blockade of its receptors provides a new and effective approach to therapy for patients with these conditions. Bosentan (Tracleer ) is the first orally-active dual endothelin receptor antagonist and has recently been approved in the US, Canada, Switzerland and the EU for the treatment of pulmonary arterial hypertension. Bosentan significantly improves exercise capacity, symptoms and functional status in patients with this disease and also slows clinical deterioration, which may be indicative of a delay of disease progression. Results from large-scale studies of bosentan in patients with pulmonary arterial hypertension and chronic heart failure have established its long-term safety and tolerability profiles. The introduction of the dual endothelin receptor antagonist bosentan has provided an essential treatment for pulmonary arterial hypertension and ongoing trials are evaluating its potential role in the management of other endothelin-mediated disease states. PMID- 12084010 TI - TACE inhibition: a new approach to treating inflammation. AB - Clinical trials showing the benefits of reducing the effects of TNF- alpha in rheumatoid arthritis have highlighted the key role of the cytokine TNF- alpha in this inflammatory condition. A new approach to reducing the effects of TNF- alpha is to decrease its synthesis by inhibiting TNF- alpha converting enzyme with GW3333. In rat models of arthritis, GW3333 has some beneficial effects. Further longer-term studies of GW3333 in animal models are required to determine whether its benefit is maintained. TACE inhibition may represent a new approach to treating inflammation. PMID- 12084011 TI - Staurosporine induces endothelial cell apoptosis via focal adhesion kinase dephosphorylation and focal adhesion disassembly independent of focal adhesion kinase proteolysis. AB - The survival of endothelial cells is dependent on interactions between the matrix and integrins mediated through focal adhesions. Focal adhesion kinase (FAK) is thought to play a key role in maintaining focal adhesion function and cell survival, whereas caspase-mediated FAK proteolysis is implicated in focal adhesion disassembly during apoptosis. We examined the relationship between changes in FAK phosphorylation and proteolysis during apoptosis of primary porcine aortic endothelial cells (PAEC) induced by staurosporine, a widely used apoptogenic agent in diverse cell types. Staurosporine-induced PAEC apoptosis was detected after 1 h and was preceded by disruption and loss of FAK localization to focal adhesions within a few minutes, whereas staurosporine-induced cleavage of FAK occurred only after 8-24 h. Staurosporine induced a very rapid dephosphorylation of FAK at Tyr(861) and Tyr(397) and caused dissociation of phosphorylated FAK from focal adhesions as early as 30 s. The effect of staurosporine was very potent with striking inhibition of Tyr(861) and Tyr(397) phosphorylation and focal adhesion disruption occurring in the range 10-100 nM. Selective inhibition of a known target of staurosporine, protein kinase C, using GF109203X, and of phosphoinositide 3'-kinase using wortmannin, did not reduce FAK tyrosine phosphorylation at Tyr(861) and Tyr(397), or cause disruption of focal adhesions. Cycloheximide, the protein synthesis inhibitor, induced PAEC apoptosis more slowly than staurosporine, but did not induce FAK dephosphorylation or rapid focal adhesion disruption, and instead caused a slower loss of focal adhesions and a marked increase in FAK proteolysis. These studies show that FAK dephosphorylation and focal adhesion disassembly are very early events mediating the onset of staurosporine-induced endothelial cell apoptosis and are dissociated from FAK proteolysis. Cycloheximide induces apoptosis through a pathway involving FAK proteolysis without dephosphorylation. PMID- 12084013 TI - Elasticity analysis and design for large metabolic responses produced by changes in enzyme activities. AB - Metabolic control analysis has been extensively used to describe how the sensitivity properties of the component enzymes in a metabolic pathway (represented by the elasticity coefficients) determine the way in which metabolic variables respond (described by the control coefficients). Similarly, metabolic control design addresses the inverse problem of obtaining the sensitivity properties of the component enzymes that are required for the system to show a pre-established pattern of responses. These formalisms, including what is called elasticity analysis and design, were developed for small, strictly speaking infinitesimal, changes. Here we extend them to large metabolic responses. The new approach can be applied to simple two-step pathways or to any arbitrary metabolic system divided into two groups linked by one intermediate. General expressions that relate control and elasticity coefficients for large changes are derived. Concentration and flux connectivity relationships are obtained. The relationships for large changes indicate that the pattern of responses is not necessarily the same as the one obtained with the traditional infinitesimal approach, in some cases the patterns being qualitatively different. The general analysis is used to study the control of ketogenesis in rat liver mitochondria, starting from data available in the literature. The control profile of the pathway subject to large changes shows both quantitative and qualitative differences from the one obtained from an analysis that is performed with infinitesimal coefficients. This exemplifies the type of errors that may be introduced when drawing conclusions about large metabolic responses from results obtained with an infinitesimal treatment. PMID- 12084012 TI - Site-directed mutagenesis reveals a novel catalytic mechanism of Mycobacterium tuberculosis alkylhydroperoxidase C. AB - Mycobacterium tuberculosis alkylhydroperoxidase C (AhpC) belongs to the peroxiredoxin family, but unusually contains three cysteine residues in its active site. It is overexpressed in isoniazid-resistant strains of M. tuberculosis. We demonstrate that AhpC is capable of acting as a general antioxidant by protecting a range of substrates including supercoiled DNA. Active site Cys to Ala mutants show that all three cysteine residues are important for activity. Cys-61 plays a central role in activity and Cys-174 also appears to be crucial. Interestingly, the C174A mutant is inactive, but double mutant C174/176A shows significant revertant activity. Kinetic parameters indicate that the C176A mutant is active, although much less efficient. We suggest that M. tuberculosis AhpC therefore belongs to a novel peroxiredoxin family and might follow a unique disulphide-relay reaction mechanism. PMID- 12084014 TI - Molecular cloning and biochemical characterization of rabbit factor XI. AB - Human factor XI, a plasma glycoprotein required for normal haemostasis, is a homodimer (160 kDa) formed by a single interchain disulphide bond linking the Cys 321 of each Apple 4 domain. Bovine, porcine and murine factor XI are also disulphide-linked homodimers. Rabbit factor XI, however, is an 80 kDa polypeptide on non-reducing SDS/PAGE, suggesting that rabbit factor XI exists and functions physiologically either as a monomer, as does prekallikrein, a structural homologue to factor XI, or as a non-covalent homodimer. We have investigated the structure and function of rabbit factor XI to gain insight into the relation between homodimeric structure and factor XI function. Characterization of the cDNA sequence of rabbit factor XI and its amino acid translation revealed that in the rabbit protein a His residue replaces the Cys-321 that forms the interchain disulphide linkage in human factor XI, explaining why rabbit factor XI is a monomer in non-reducing SDS/PAGE. On size-exclusion chromatography, however, purified plasma rabbit factor XI, like the human protein and unlike prekallikrein, eluted as a dimer, demonstrating that rabbit factor XI circulates as a non-covalent dimer. In functional assays rabbit factor XI and human factor XI behaved similarly. Both monomeric and dimeric factor XI were detected in extracts of cells expressing rabbit factor XI. We conclude that the failure of rabbit factor XI to form a covalent homodimer due to the replacement of Cys-321 with His does not impair its functional activity because it exists in plasma as a non-covalent homodimer and homodimerization is an intracellular process. PMID- 12084017 TI - Outcomes of day care: a pilot study on changes in cognitive function and agitated behaviors of demented elderly in Korea. AB - This study examined outcomes of Korean day care for demented elderly Koreans. Changes in the cognitive function and agitation of demented elderly individuals were assessed over time. A one-group repeated measures design was used. The data were collected on a sample of 13 individuals from March to August 1999. The cognitive function of the subjects remained stable during the 10-week study period, while their level of agitation increased over time, indicating a need for the examination of factors that affect problematic behaviors of the demented elderly. This pilot study showed that day-care programs in Korea may be an effective way to maintain the stability of cognitive function in elderly Koreans with mild- to moderate-stage dementia. Further investigation of effective behavioral modifications within day-care settings could justify additional government support of Korean day-care programs for those suffering from dementia. PMID- 12084019 TI - Using a model to evaluate nursing education and professional practise. AB - The concept of evaluation is becoming increasingly ambiguous and a lot of processes may be called evaluation without any clear definitions. A theoretical frame of reference may function as a compass in an evaluation context when collecting, analysing and interpreting data as well as drawing conclusions. The purpose of the present study was to present and discuss the applicability of an educational interaction model for the evaluation of nursing education programs and the professional competence of nurses. The model combines different dimensions in the educational process, using both a student and an educational perspective. It is not uncommon for evaluations to concentrate on one dimension only, which tends to give an insufficient picture of the process of interaction. Examples are provided from nurse students/nurses education and professional practise to show that the relationship between students' abilities and educational factors, in the form of intentional goals and educational frameworks, have an influence on educational outcome. PMID- 12084018 TI - Self-esteem and student nurses: a cross-cultural study of nursing students in Thailand and the UK. AB - Self-esteem is a key feature in a person's perception of their own worth. This report is of a study of the reported self-esteem levels of two groups of student nurses: one in Thailand and one in the UK. Purposive samples of 120 Thai students and 101 UK undergraduate nursing students were given the Culture-Free Self-Esteem Inventory (CFSEI-2). The CFSEI-2 is a self-reported inventory, which measures an individual's perception of self. The findings of the study indicate that the perceptions of own self-esteem in undergraduate student nurses in the UK and in Thailand were comparable to the normal ranges of self-esteem as assessed by the instrument. An independent sample t-test revealed that there were no significant differences in mean overall and subscale self-esteem scores between UK and Thai nursing students. There were no indications of differences in levels of self esteem for UK and Thai nursing students experiencing different parts of their training. PMID- 12084015 TI - Structural and functional characterization of the USP11 deubiquitinating enzyme, which interacts with the RanGTP-associated protein RanBPM. AB - RanBPM is a RanGTP-binding protein required for correct nucleation of microtubules. To characterize the mechanism, we searched for RanBPM-binding proteins by using a yeast two-hybrid method and isolated a cDNA encoding the ubiquitin-specific protease USP11. The full-length cDNA of USP11 was cloned from a Jurkat cell library. Sequencing revealed that USP11 possesses Cys box, His box, Asp and KRF domains, which are highly conserved in many ubiquitin-specific proteases. By immunoblotting using HeLa cells, we concluded that 921-residue version of USP11 was the predominant form, and USP11 may be a ubiquitous protein in various human tissues. By immunofluorescence assay, USP11 primarily was localized in the nucleus of non-dividing cells, suggesting an association between USP11 and RanBPM in the nucleus. Furthermore, the association between USP11 and RanBPM in vivo was confirmed not only by yeast two-hybrid assay but also by co immunoprecipitation assays using exogenously expressed USP11 and RanBPM. We next revealed proteasome-dependent degradation of RanBPM by pulse-chase analysis using proteasome inhibitors. In fact, ubiquitinated RanBPM was detected by both in vivo and in vitro ubiquitination assays. Finally, ubiquitin conjugation to RanBPM was inhibited in a dose-dependent manner by the addition of recombinant USP11. We conclude that RanBPM was the enzymic substrate for USP11 and was deubiquitinated specifically. PMID- 12084020 TI - Family support and self-care behavior of Chinese chronic obstructive pulmonary disease patients. AB - A descriptive correlation study was conducted at the outpatient pulmonary clinic of a large medical center to describe family support and self-care behavior and to ascertain the relationship of these two variables among Chinese chronic obstructive pulmonary disease (COPD) patients. A total of 98 COPD patients completed the questionnaires providing demographic data and information about perceived family support and self-care behavior. Results showed that most subjects perceived that they had received a high level of family support, and the majority of the subjects perceived that they had initiated an adequate amount of self-care behavior. A slightly statistically significant positive relationship between family support and self-care behavior was ascertained. PMID- 12084022 TI - COHb: a stress-induced marker reflecting surgical insults? PMID- 12084021 TI - Evaluation of a training program for the prevention of lower back pain among hospital employees. AB - An evaluation of a training program for the prevention of lower back pain (LBP) was conducted over a two-year period in the Regional Hospital of Le Mans, which is located in the west of France and employs 2400 staff. The training program was based on a multidisciplinary approach to worker training in the prevention of LBP and in ergonomic efforts in the workplace. Workers received theoretical training on LBP, patient handling and ergonomics, and on-the-job ergonomics instruction to help them to implement participative ergonomics in their working groups. A cohort of 136 employees attended classes on safe posture and patient handling, then received advice from instructors who observed them while they performed their typical workday tasks. PMID- 12084023 TI - Pediatric liver disease in the United States: epidemiology and impact. AB - Liver diseases which affect children are unique. Diseases such as inborn errors of metabolism are fertile grounds for investigation. Study of these 'experiments of nature' will provide insight into hepatobiliary function and lead to novel treatment modalities. Successful treatment or prevention of liver disease in children will have long-term implications. This report describes the current burden of pediatric liver disease and offers approaches to intervention. PMID- 12084024 TI - Non-cirrhotic portal fibrosis: current concepts and management. AB - Non-cirrhotic portal hypertension (NCPH) comprises diseases having an increase in portal pressure (PP) due to intraheptic or prehepatic lesions, in the absence of cirrhosis. The lesions are generally vascular, either in the portal vein, its branches or in the perisinusoidal area. Because the wedged hepatic venous pressure is near normal, measurement of intravariceal or intrasplenic pressure is needed to assess PP. The majority of diseases included in the category of NCPH are well-characterized disease entities where portal hypertension (PHT) is a late manifestation and, hence, these are not discussed. Two diseases that present only with features of PHT and are common in developing countries are non-cirrhotic portal fibrosis (NCPF) and extrahepatic portal vein obstruction (EHPVO). Non cirrhotic portal fibrosis is a syndrome of obscure etiology, characterized by 'obliterative portovenopathy' leading to PHT, massive splenomegaly and well tolerated episodes of variceal bleeding in young adults from low socioeconomic backgrounds, having near normal hepatic functions. In some parts of the world, NCPF is called idiopathic portal hypertension (IPH) or 'hepatoportal sclerosis'. Because 85-95% of patients with NCPF and EHPVO present with variceal bleeding, treatment involves management with endoscopic sclerotherapy (EST) or variceal ligation (EVL). These therapies are effective in approximately 90-95% of patients. Gastric varices are another common cause of upper gastrointestinal bleeding in these patients and these can be managed with cyanoacrylate glue injection or surgery. Other indications for surgery include failure of EST/EVL, and symptomatic hypersplenism. The prognosis of patients with NCPF is good and 5 years survival in patients in whom variceal bleeding can be controlled has been reported to be approximately 95-100%. PMID- 12084025 TI - Perioperative changes in carbonylhemoglobin and methemoglobin during abdominal surgery: alteration in endogenous generation of carbon monoxide. AB - BACKGROUND: Carbon monoxide (CO), which is homologous to nitric oxide (NO) as a monoxide, has been recently studied as a novel gaseous mediator for the maintenance of circulatory homeostasis and as a regulator of organ functions. Abdominal surgery is supposed to modulate the gaseous mediator by the reduction of heme oxygenase (HO) activity or transcriptional regulation of inducible HO. Therefore, we investigated perioperative changes in CO generation during abdominal surgery. METHODS: A total of 397 patients who received abdominal surgery under intubation anesthesia were studied retrospectively by spectrophotometric analysis of carbonylhemoglobin (COHb) and methemoglobin (metHb) in arterial blood obtained at three points: before operation; several hours after return from operation room (0 POD); and next morning (1 POD). Thirty three splenectomies, 36 hepatectomies and 42 drainages with control of infection focus for peritonitis were compared with 286 controls. The influences of smoking and blood transfusion were disregarded in the analysis, because smoking and blood transfusion were shown to increase exogenous and endogenous CO, respectively. RESULTS: In the non-smoker control group without blood transfusion, COHb did not change during the perioperative period, while metHb increased from the preoperative value of 0.52 +/- 0.03 to 0.72 +/- 0.02 at 0 POD and returned to 0.45 +/- 0.03 mg/dL at 1 POD. In the splenectomy group, COHb decreased from the preoperative value of 1.63 +/- 0.36 to 1.19 +/- 0.20 and 1.13 +/- 0.26 mg/dL at 0 and 1 POD, respectively, as a result of the removal of the organ with high HO activity. In the splenectomy group MetHb remained low: 0.47 +/- 0.09 mg/dL at 0 POD as compared with the control value. In the peritonitis and hepatectomy groups, COHb did not change during the perioperative period, while metHb increased to 0.64 +/- 0.06 and 0.73 +/- 0.10 mg/dL at 1 POD, respectively, as compared with the control value. In the hepatectomy group with or without blood transfusion, however, COHb and metHb were higher at 1 POD than the corresponding control value. CONCLUSION: Changes in COHb and metHb concentrations in arterial blood occur during abdominal surgery, although these amplitudes are small when compared with CO intoxication and methemoglobulinemia. It is likely that organ perfusion and functions are affected by these monoxide gas mediators during abdominal surgery. PMID- 12084026 TI - Newly developing hiatus hernia: a survey in patients undergoing upper gastrointestinal endoscopy. AB - BACKGROUND: Data on the incidence of hiatus hernia are lacking. A cross-sectional study was performed in a large population of consecutive patients undergoing endoscopy in order to assess the yearly incidence of hiatus hernia in this population. METHODS: Consecutive patients undergoing upper gastrointestinal endoscopy in whom no macroscopic abnormalities were seen and who, in addition, underwent a second endoscopy were included in the study. The presence of newly developed hiatus hernia was noted, as well as the time elapsed between both endoscopies. RESULTS: Over a period of 8 years, 12 122 endoscopies were performed in 9580 patients. Ninety patients developed a hiatus hernia; this was not the case in a control group of 353 patients. Patients who developed a hiatus hernia were significantly older than those who did not (P < 0.001). The number of women who developed hiatus hernia was higher than the number of men who developed hiatus hernia (P < 0.0001). The total time between both endoscopies in 443 patients was 897 patient years. Ninety patients (20.3%) developed a hiatus hernia. If these data are extrapolated to a yearly occurrence, then 35 of 176 patients will develop a hiatus hernia. It takes an average of 1.9 years for a hiatus hernia to develop. CONCLUSIONS: In the present study, it was calculated that 19.9% of the studied population would develop a hiatus hernia per year. Patients who developed a hiatus hernia were significantly older than patients who did not. The present study also shows that a hiatus hernia actually develops later in life. PMID- 12084027 TI - The Hong Kong index of dyspepsia: a validated symptom severity questionnaire for patients with dyspepsia. AB - AIM: Locally validated symptom questionnaires are essential instruments in dyspepsia research. A symptom score for Chinese dyspeptics was developed. Multiple aspects of validity and reliability were assessed. METHODS: Sixty-five patients with endoscopy-negative dyspepsia and 65 healthy controls were presented with a list of 24 previously determined common abdominal symptoms and asked to rate the severity in a five point Likert scale. Comprehensibility and relevance of symptoms were assessed. The questionnaire was administered 3 h later to assess for reproducibility. A locally validated Short Form-36 quality-of-life questionnaire was also administered. Cisapride 5 mg thrice daily was prescribed to dyspeptic patients but not to the controls. The dyspepsia symptom and SF-36 questionnaires were repeated after 3 weeks for patients under treatment. Global change in symptoms and the need for further medical care were assessed. Concept, content, construct and criterion validity, consistency and reproducibility of the dyspepsia symptom questionnaire were assessed. RESULTS: All items were considered comprehensible by more than 90% of subjects. Relevance of individual symptoms to dyspeptic patients ranged from 10.8-76.9%. Twelve items were selected by logistic regression to account for most of the differences between control and dyspeptic patients. Test-retest reproducibility and internal consistency were good with the intraclass correlation coefficient of 0.89 and Cronbach's alpha coefficient of 0.90. A cut-off score of equal to or greater than 16 was determined to discriminate between controls and dyspeptic patients. The dyspepsia score correlated negatively with all domains of the SF-36 quality of life scale except physical functioning. The dyspepsia questionnaire also discriminated between patients who reported a subjective improvement in symptoms and those who reported no change or worsening. CONCLUSIONS: The dyspepsia symptom index was easy to understand, internally consistent and reproducible. It predicted global symptom change, and the symptom severity scores correlated negatively with quality of life. PMID- 12084028 TI - Chinese tea consumption and lower risk of Helicobacter infection. AB - BACKGROUND: Our first study had shown that Chinese tea has anti-Helicobacter activity. The present study investigated the relationship between patients'Helicobacter status and the history of Chinese tea consumption. METHODS: Chinese patients who had their first upper endoscopy were recruited. Before the procedure, patients completed questionnaires about their current Chinese tea consumption habits and those when they were around 10-25 years old. This information was used to calculate the tea consumption indices (TCI). Gastric biopsies (two from the antrum and two from the corpus) were taken for histological examination for Helicobacter. RESULTS: Eighty patients were recruited. Eight patients were rejected because of either an incomplete questionnaire, absence of gastric biopsy or a suspected history of treatment for Helicobacter infection. Of the 72 patients, 42 (58.3%) were Helicobacter positive. The age, sex ratio and indications for endoscopy were similar in both Helicobacter-positive and -negative groups. Classifying patients using either current, past or total TCI, patients with high tea consumption had significantly lower Helicobacter infection rate than those with low tea consumption (45% compared to 74%, 42% compared to 67% and 40% compared to 83%, respectively). Helicobacter-negative patients had higher overall TCI than Helicobacter-positive patients (28.2:17.3). When current and previous histories were analyzed separately, Helicobacter-negative patients also had a trend towards higher TCI currently (16.6:11.0) and in their 20s (11.6:6.1). CONCLUSIONS: There is a significant inverse relationship between Chinese tea consumption and Helicobacter infection. Chinese tea consumption may decrease the chance of Helicobacter infection. PMID- 12084029 TI - Relationship between nuclear factor-kappaB activation and virulence factors of Helicobacter pylori in Japanese clinical isolates. AB - BACKGROUND: Several factors have been proposed as a possible virulence determinant of Helicobacter pylori infection. The aim of the present study was to evaluate these candidates in nuclear factor (NF)-kappaBeta activation, which is a critical regulator of genes involved in inflammatory reactions. METHODS: We determined the status of cagE, iceA, HP0441 (a virB4 homolog), the s1 signal sequence of vacA and babA2 by polymerase chain reaction, all of which are candidate virulence determinants, in 107 H. pylori strains isolated from Japanese patients. Nuclear factor-kappaBeta activation was evaluated by the luciferase reporter assay. The gastric mucosa of the hosts was examined histologically. RESULTS: The cagE gene was positive in 102 (95.3%) strains, iceA1 was positive in 71 (66.4%) strains, HP0441 was positive in 68 (63.6%) strains, vacA s1 was positive in 105 (98.1%) strains and babA2 was positive in 103 (96.3%) strains. Nuclear factor-kappaBeta was activated by all cagE-positive strains, but was not activated by any of the cagE-negative strains. The status of iceA or HP0441 was not associated with NF-kappaBeta activation. Neutrophil infiltration in gastric mucosa was significantly more severe in patients infected with cagE-positive strains than in patients infected with negative strains. No association was found between the degree of neutrophil infiltration and the status of HP0441 or iceA. Due to very high positivity of vacA s1 and babA2 in Japanese strains, their roles remain to be investigated. CONCLUSIONS: The cag pathogenicity island (PAI) status, as determined by cagE polymerase chain reaction, but not the status of iceA or HP0441, is closely associated with NF-kappaBeta activation and the degree of gastric mucosal inflammation in the hosts. PMID- 12084030 TI - Decreased production of insulin-like growth factor-binding protein (IGFBP)-6 by transfection of colon cancer cells with an antisense IGFBP-6 cDNA construct leads to stimulation of cell proliferation. AB - BACKGROUND: : Previously, we have observed that highly unsaturated dietary (n-3) fatty acids inhibit cell proliferation in conjunction with stimulation of insulin like growth factor-binding protein (IGFBP)-6 secretion in Caco-2 cells, a human colon carcinoma cell line. METHODS: : To test the converse hypothesis that inhibition of endogenous IGFBP-6 secretion stimulates Caco-2 cell proliferation, cells were transfected with the antisense IGFBP-6 expression construct or pcDNA3 vector only, and single colonies resistant to G418 sulfate were isolated. RESULTS: : Our initial studies indicated that three antisense clones grew faster and produced less IGFBP-6 than two pcDNA3 clones, so antisense IGFBP-6 #5 and pcDNA3 #8 were selected for further detailed analysis. Both the control and antisense clones grew in serum-free medium reaching a plateau density at day eight. However, the antisense clone grew at a rate faster than that of the control and reached a final density that was 31 +/- 3% higher than the control. Northern blot, ligand blot and immunoblot analyses revealed that accumulation of IGFBP-6 mRNA and concentrations of IGFBP-6 peptide produced by the antisense clone were decreased by 80-90% compared to the control. The doubling times of the antisense and control clones were 21.9 +/- 0.4 and 24.8 +/- 0.3 h (P < 0.05), respectively. Exogenous IGF-I and IGF-II (0.2-200 nmol/L) stimulated proliferation of both the control and antisense clones in a dose-dependent manner, but the relative potency and efficacy of IGF-II was higher in the antisense clone compared to the control. These results indicate that suppression of IGFBP-6 secretion correlates with an increase in the basal rate of Caco-2 cell growth. CONCLUSIONS: : Our findings are consistent with the hypothesis that IGFBP 6 inhibits cell growth by binding to endogenously produced IGF-II, thereby preventing IGF-II from interacting with the IGF-I receptor to stimulate cellular proliferation by an autocrine mechanism. PMID- 12084031 TI - Long-term quality of life of postoperative rectal cancer patients. AB - BACKGROUND: The long-term quality of life (QOL) of postoperative rectal cancer patients has not been previously investigated in Japan and may vary depending on the surgical technique used (i.e. with or without a stoma). METHODS: The Kanagawa Cancer Registry was used to select 348 rectal cancer patients who underwent surgery at the St Marianna University Hospital between 1978 and 1997. Of these, 164 surviving patients were sent a postal survey consisting of the Japanese EuroQol instrument with an additional questionnaire on present symptoms and lifestyle. RESULTS AND CONCLUSIONS: One hundred and ten responses were received, including 38 from stoma patients. The presence of a stoma did not affect the QOL of the male patients, while it did affect that of the female patients. The QOL of the long-term survival group was associated with several items concerning lifestyle, symptoms and usual activity, and the association did not depend on the presence of a stoma. The long-term QOL could be recognized according to the characteristics of rectal cancer patients, independent of the presence of a stoma. PMID- 12084032 TI - High sustained response to daily dosing of interferon with ribavirin in chronic hepatitis C patients naive to therapy. AB - BACKGROUND: : Viral kinetics suggests that daily administration of alpha interferon (IFN) will clear hepatitis C virus (HCV) RNA earlier and more frequently compared with standard t.i.w. To reduce the likelihood of viral replication, mutation and subsequent development of resistance, daily dosing with IFN may be appropriate. To determine the safety and efficacy of daily IFN with ribavirin in chronic HCV infection we performed a prospective study. METHODS: : Thirty-five naive adult HCV-positive patients (25 male/10 female) were treated with IFN-alpha2b; 5 MU daily for 2 weeks followed by 3 MU daily for 22 weeks and ribavirin 800-1200 mg/day depending on weight. Liver biopsy, performed in 25 patients, showed mild to moderate activity in 19 patients (76%) and severe activity in six patients (24%). Two patients showed staged IV fibrosis. Serotyping was performed in 29 patients by an enzyme immunoassay-based Murex assay. Type 3 was the predominant serotype, present in 14 cases. Hepatitis C virus RNA was measured by the Chiron bDNA assay. RESULTS: : Mean baseline HCV-RNA level was 14.2 +/- 18.7 MEq/mL (median 6.09; range 0.2-92.5), which became undetectable in all but three patients at week 4. Normalization of alanine aminotransferase (ALT) at week 4 was seen in 27 patients. Three patients withdrew due to non-compliance. Thirty-two patients completed 24 weeks of therapy as per the protocol. At the end of treatment, the HCV-RNA level was negative in 29 of 32 patients (90.6%) and ALT was normal in 31 of 32 patients (97%). Sustained viral response at 6 months follow up was seen in 28 of 32 patients (88%). The ALT level was normal in 28 of 32 patients (88%). CONCLUSION: : Daily administration of IFN with ribavirin is well tolerated in the majority of patients. There is rapid elimination of virus with normalization of ALT and a significantly high sustained viral response. PMID- 12084033 TI - Chronic liver disease mortality attributable to hepatitis B and C in New Zealand. AB - BACKGROUND: This research aimed to estimate the prevalence and population attributable risk percent (PAR%) of hepatitis B (HBV) and C (HCV) infection among chronic liver disease (CLD) deaths in New Zealand. The PAR% is the percentage of CLD cases attributable to either HBV or HCV. Within New Zealand, there are large differences in HBV prevalence by ethnic group, so prevalence and PAR% estimates were made separately for the three major ethnic groups. METHODS: The study sample was selected from CLD deaths between 1992 and 1997. Data were extracted from hospital records and coroners' reports. The prevalence and PAR% of HBV and HCV were estimated. RESULTS: Data were extracted for 303 of 359 decedents selected for inclusion. Hepatitis B virus and HCV test results were identified in 67 and 43%, respectively. Among those cases tested, the prevalence (and estimated PAR%) of HBV infection was 68% (PAR% 66%) for Pacific people, 54% (PAR% 52%) for Maori and 10% (PAR% 10%) for European New Zealanders. The prevalence (and estimated PAR%) of past or present HCV infection ranged between 8 and 15% (PAR% 8-14%) for the three major ethnic groups. CONCLUSIONS: The present study has demonstrated that HBV and HCV infections are important contributors to CLD mortality in New Zealand. With the introduction of universal hepatitis B vaccination in the late 1980s, we would expect the burden of CLD deaths attributable to HBV to decrease in the future. However, the burden of CLD deaths due to HCV is likely to increase. PMID- 12084035 TI - Hepatic Kupffer cell phagocytotic function in rats with erythrocytic-stage malaria. AB - BACKGROUND: In the erythrocytic phase of malaria, Kupffer cells show marked hypertrophy and hyperplasia and are filled with malarial pigment. However, phagocytic function in this state has not been well characterized. The aim of the present study was to use mouse Plasmodium berghei to infect rats with malaria and study the phagocytic function and morphology of Kupffer cells. METHODS: We used a recirculating isolated perfused rat liver (IPRL) to quantitate Kupffer cell phagocytic clearance of radiolabeled albumin-latex over 120 min in high parasitemia (53 +/- 6%; n = 7) and low parasitemia (approximately 1%; n = 4) malaria-infected rats and littermate controls (n = 7 and n = 4, respectively). In a further group of high-parasitemic rats, perfusion was ceased after 7 min and liver radioactivity also measured. Electron microscopy was performed after perfusions. RESULTS: In high-parasitemia malaria rats, clearance of radiolabeled latex from IPRL perfusate over 120 min was significantly (P < 0.01) faster than in controls, with a lower area under the curve (0.19 +/- 0.02 vs 0.43 +/- 0.07 /mL per min, respectively) and shorter half-life (t1/2k; 2.4 +/- 0.6 vs 10.0 +/- 2.3 min, respectively). Low-parasitemia rats were identical to controls. After 7 min perfusion in high-parasitemic rats (n = 4), total radioactivity in liver homogenates was higher than in controls (n = 4; 33.1 +/- 6.2 vs 18.4 +/- 1.9% of injected radiolabel; P < 0.05). Electron microscopy showed latex in Kupffer cells, more abundantly seen in high-parasitemic animals. CONCLUSIONS: Total Kupffer cell phagocytic activity of the liver is markedly increased in rats with a high parasitemic load of malarial P. berghei infection. This is presumed to reflect an upregulation of scavenger activity phagocytosing erythrocytes and their breakdown products. PMID- 12084034 TI - Accelerated reversal of carbon tetrachloride-induced cirrhosis in rats by the endothelin receptor antagonist TAK-044. AB - BACKGROUND: A multifunctional mediator, endothelin (ET)-1 is implicated in the pathophysiology of liver cirrhosis. Carbon tetrachloride (CCl4)-induced cirrhosis in rats resolves upon termination of CCl4 treatment. We determined the hepatic ET 1 system during such reversal and assessed whether ET-1 receptor antagonism enhances this process. METHODS: Cirrhosis was induced in rats by CCl4 treatment for 8 weeks. Treatment with an ETA/ETB antagonist TAK-044 (10 mg/kg per day) was then started and determinations were made at 1, 2 and 4 weeks. RESULTS: After termination of CCl4 treatment, accelerated normalization of liver architecture and portal hypertension occurred in TAK-044-treated rats compared with saline treated rats. The increased hepatic hydroxyproline concentration and collagen I mRNA expression also declined to greater extents in the TAK-044-treated group. Higher collagenase activity in cirrhosis decreased in saline-treated rats, but did not reach basal values. In TAK-044-treated rats, collagenase activity tended to increase at weeks 2 and 4. Increased ET-1 concentration and ETA receptor density declined to normal values in both groups. In contrast, increased ETB receptor density did not change in saline-treated rats, but decreased to control values in TAK-044-treated rats. CONCLUSIONS: Our results emphasize the role of ET 1 in chronic liver disease and strongly indicate the potential for ET-1 receptor antagonists in its treatment. PMID- 12084037 TI - Different outcomes of vertical transmission of hepatitis C virus in a twin pregnancy. AB - The risk of vertical transmission of hepatitis C virus (HCV) from mother to infant has been well documented, but the exact mode of transmission is still unclear. In a set of monochorionic diamniotic monozygous twins, only the second baby was infected with HCV from the mother who was positive for serum HCV-RNA. The babies had an uncomplicated vaginal delivery 3 min apart and they were both bottle fed from the outset. The second baby developed clinical hepatitis that persisted to 30 months follow up. The intrauterine environment should have been identical for these twins, and therefore, the maternal HCV factors, including viral load are not the sole determining factors for mother-to-infant transmission of HCV. PMID- 12084036 TI - Xanthine oxidase-derived oxygen radicals play significant roles in the development of chronic pancreatitis in WBN/Kob rats. AB - BACKGROUND: Although oxygen-derived free radicals are known to play a role in cell injury and DNA alterations, the role of active oxidants in chronic pancreatitis has not been fully elucidated. Using WBN/Kob rats, which spontaneously develop chronic pancreatitis-like lesions, we investigated whether xanthine oxidase (XOD)-derived oxygen radicals are involved in pancreatic tissue injury. METHODS: WBN/Kob rats were fed a control or a tungsten diet. The latter depletes XOD activity. Histologic al changes, glutathione (GSH) content and XOD and superoxide dismutase (SOD) activities were determined in pancreatic tissue. Pancreatic 8-hydroxy-deoxyguanosine (8-OH-dG) levels and lithostathine mRNA were also examined. RESULTS: In WBN/Kob rats, parenchymal destruction and fibrosis developed at approximately 12 weeks of age and progressed with each month. The activity of XOD was significantly higher in the early period (8-12 weeks), whereas the levels of GSH and SOD decreased after 16 weeks. Levels of 8-OH-dG in WBN/Kob rats were significantly elevated at 16 weeks. Lithostathine mRNA levels started to increase at 8 weeks, but were suppressed at 16 weeks. The tungsten diet significantly attenuated the histological changes in WBN/Kob rats. The increase in pancreatic XOD activity and 8-OH-dG content in WBN/Kob rats was significantly inhibited by the tungsten diet and lithostathine mRNA levels remained high at 16 weeks. CONCLUSION: These results suggest that oxygen radicals generated by XOD play an important role in oxidative DNA damage and the development of chronic pancreatic injury. PMID- 12084038 TI - Gastrointestinal: vascular lesions of the stomach. PMID- 12084039 TI - Hepatobiliary and pancreatic: choledochoduodenostomy. PMID- 12084040 TI - Iron-deficiency anemia complicating AL amyloidosis with recurrent small bowel pseudo-obstruction and hindgut sparing. AB - AL amyloidosis complicating multiple myeloma is rare but well recognized. Endoscopic appearances of amyloid in the gastrointestinal tract can, however, be highly variable. Macroscopic appearances of the duodenum can range from scalloping (often seen in celiac disease) to that of duodenitis; erosive disease, frank ulceration and even protruberant masses. The importance of small bowel biopsies at the time of duodenal intubation and in gastroscopic examinations for iron deficiency anemia, must therefore be stressed. PMID- 12084041 TI - Fulminant hepatic failure associated with benzbromarone treatment: a case report. PMID- 12084042 TI - Effect of undernourishment on Herpes Simplex Virus Type 1 ocular infection in the Wistar rat model. AB - We have studied the susceptibility to Herpes Simplex Virus Type 1 (HSV-1) infection in malnourished rats. Groups of 10 rats were undernourished during suckling by offspring duplication. The animals were put on commercial diet and at 1, 2, 3, 5 and 8 weeks after weaning, infected in the eye by scarification with HSV-1, strain F. Significant differences in morbidity and mortality were observed between malnourished and control groups infected three weeks after weaning. Viral titres were higher in ocular washings and brains obtained from the malnourished group. This group showed a diminution in antigen dependent lymphocyte proliferation compared to control, and significantly lower delayed type hypersensitivity reaction against inactivated virus (malnourished = 0.16 +/- 0.02 mm, control = 0.26 +/- 0.03 mm, p < 0.05). Neutralizing antibodies in serum were lower in the malnourished group and lower levels of interferon were obtained in the malnourished group 24 h post-infection. We conclude that malnutrition during suckling induces a delay in the capability to overcome HSV infection. PMID- 12084043 TI - Evidence for a perforin-mediated mechanism controlling cardiac inflammation in Trypanosoma cruzi infection. AB - CD8+ T lymphocytes are considered an important cell population involved in the control of parasitaemia and mortality after Trypanosoma cruzi infection. However, despite recent developments in this field, the mechanism whereby this control is exerted is still not completely understood. Here we have used perforin knockout ( /-) mice infected with Y strain T. cruzi in order to evaluate specifically the participation of the perforin-based cytotoxic pathway in the destruction of cardiomyocytes, cellular inflammatory infiltration, and control of parasitaemia and mortality. We observed that although parasitaemia was equivalent in perforin (+/+) and (-/-) groups, survival rate and spontaneous physical performance were significantly lower in the perforin deficient mice. The cardiac inflammatory cell infiltration, mostly composed of CD8+ cells, was more evident in perforin (-/-) mice. Ultrastructural and immunofluorescence analysis, as well as plasma creatine kinase activity, revealed cardiomyocyte damage and necrosis, more evident in perforin (-/-) mice. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assays performed in heart samples revealed similar and modest levels of apoptosis in both perforin (+/+) and (-/-) mice. These results indicate that perforin does not play a pivotal role in the control of parasitaemia and direct lysis of cardiomyocytes, but seems to be an important molecule involved in the control of cardiac inflammation and pathology induced by a highly virulent strain of T. cruzi. PMID- 12084044 TI - Systemic administration of interferon-gamma-expressing plasmid reduces late allergic bronchitis in a mouse model of asthma. AB - Asthma might be caused by a helper T(Th)2 immune response. We hypothesized that the systemic administration of the Th1 cytokines may reduce the Th2 type late asthmatic response (LAR). We examined the effect of the intraperitoneal injection of interferon(IFN)-gamma-expressing plasmid, a Th1 cytokine, or interleukin(IL)-4 expressing plasmid, a Th2 cytokine, at the time of sensitization on a mouse model of asthma induced by ovalbumin in BALB/c mice. We demonstrated that the IFN-gamma expressing plasmid reduced the LAR, whereas the IL-4-expressing plasmid enhanced the LAR as compared with the saline or plasmid-only treated group. The present study suggests that the systemic administration of IFN-gamma-expressing plasmid may have a modulating ability of Th1/Th2 balance to down-regulate Th2 response by a mutual inhibitory mechanism between Th1 and Th2 cells, leading to the reduction of the LAR. PMID- 12084045 TI - Is IL12 necessary in experimental hypersensitivity pneumonitis? AB - Inhalation of Saccharopolyspora rectivirgula (SR) can cause the disease Farmer's Lung, a classic example of hypersensitivity pneumonitis. Th1, but not Th2, cell lines can adoptively transfer experimental hypersensitivity pneumonitis (EHP). Substantial amounts of IL12 appear in bronchoalveolar lavage fluid (BALF) after a single intratracheal (IT) injection of SR, and SR-induced IL12 secretion by both a macrophage cell line and alveolar macrophages. We tested the hypothesis that IL12 is essential for the development of EHP by addition of anti-IL12 to cultured cells, and adoptive transfer of EHP in IL12p40-/- animals. We transferred SR cultured spleen and lung associated lymph node cells from SR sensitized mice (both IL12p40-/- and wild type), to naive recipients (both wild type and IL12p40 /-). The addition of anti-IL12 to cultures of sensitized cells could not ablate the ability of these cells to transfer EHP. Cultured cells from IL12p40-/- animals were fully capable of transferring EHP. In contrast, IL12p40-/- recipients of both wild type and IL12p40-/--cultured cells were less able to express EHP (lung histology and BALF characteristics) than wild type mice, and had more eosinophils in both lung tissue and BALF. We conclude that IL12 is not necessary for development of cells able to adoptively transfer EHP, but that it is required for full expression of EHP in recipient animals. PMID- 12084046 TI - Antiproliferative effect of thalidomide alone and combined with carmustine against C6 rat glioma. AB - Thalidomide could have therapeutic applications in neoplasms and in other diseases, particularly those of autoimmune origin. The objective of this study was to investigate the effect of various doses of thalidomide on the growth of C6 glioma in rats, and to determine its effects on parameters of cell proliferation and angiogenesis. Additionally, we investigated a potential enhancement of the antitumoral action of thalidomide when combined with a low dose of the antineoplastic carmustine. C6 glioma cells were implanted subcutaneously in Wistar rats. A highly malignant glioma developed in 80% of animals. When the tumour reached 2.0 cm diameter thalidomide was administered at doses of 100, 200 or 400 mg/kg/day. When given at a dose of 400 mg/kg/day thalidomide significantly reduced the tumour volume, the mitotic index and cell proliferation but not the vascular density. The combination of thalidomide plus carmustine increased the inhibitory effect on tumoral growth. Our results indicate that thalidomide is effective against malignant glioma; apparently by an antiproliferative effect, rather than by inhibition of angiogenesis; when combined with carmustine it could increase the response of glioma to antineoplastic treatment. PMID- 12084047 TI - Influence of the electric charge of the antigen and the immune complex (IC) lattice on the IC activation of human complement. AB - In order to understand the mechanism of complement (C) activation by immune complexes (ICs), the anti-complementary effect of ICs containing cationized antigens was compared in vitro to that using ICs formed by native antigens. ICs were prepared with affinity-purified rabbit polyclonal IgG antibovine serum albumin (BSA) antibody and either native BSA (isoelectric point 4.2) or BSA rendered cationic by treatment with ethylenediamine (isoelectric point 9.4). Native and cationized antigens were characterized by isoelectric focusing. ICs containing anti-BSA IgG or F(ab')2, formed either at equivalence or in excess of native or cationized antigen, were submitted to ultracentrifugation in a sucrose gradient for mesh size determination. The anti-complementary effect of ICs was evaluated by kinetic determination of haemolytic activity of human serum on haemolysin-sensitized sheep red blood cells. In conditions of antigen excess, the ICs formed by cationized BSA were significantly more efficient in activating human complement than those formed by native antigen. This higher activity was dependent on cationized antigen complexed with complete antibody molecules, as non-complexed cationized BSA or ICs prepared with F(ab')2 fragments were inactive under the same experimental conditions. Furthermore, this difference did not depend on the mesh size of the immune complexes. Our results suggest that the balance between antigen, antibody and C may be of importance in vivo for the onset and course of infections and other pathological processes involving IC formation. ICs containing cationized antigens should be proven of value in experimental models for studies on the regulation of C activation. PMID- 12084049 TI - A new conceptual framework for enzyme catalysis. Hydrogen tunnelling coupled to enzyme dynamics in flavoprotein and quinoprotein enzymes. AB - Recent years have witnessed high levels of activity in identifying enzyme systems that catalyse H-transfer by quantum tunneling. Rather than being restricted to a small number of specific enzymes as perceived initially, it has now become an accepted mechanism for H-transfer in a growing number of enzymes. Furthermore, H tunneling is driven by the thermally induced dynamics of the enzyme. In some of those enzymes that break stable C-H bonds the reaction proceeds purely by quantum tunneling, without the need to partially ascend the barrier. Enzymes studied that fall into this category include the flavoprotein and quinoprotein amine dehydrogenases, which have proved to be excellent model systems. These enzymes have enabled us to study the relationship between barrier shape and reaction kinetics. This has involved studies with "slow" and "fast" substrates and enzymes impaired by mutagenesis. A number of key questions now remain, including the nature of the coupling between protein dynamics and quantum tunneling. The wide ranging implications of quantum tunneling introduce a paradigm shift in the conceptual framework for enzyme catalysis, inhibition and design. PMID- 12084050 TI - Barrier passage and protein dynamics in enzymatically catalyzed reactions. AB - This review describes studies of particular enzymatically catalyzed reactions to investigate the possibility that catalysis is mediated by protein dynamics. That is, evolution has crafted the protein backbone of the enzyme to direct vibrations in such a fashion to speed reaction. The review presents the theoretical approach we have used to investigate this problem, but it is designed for the nonspecialist. The results show that in alcohol dehydrogenase, dynamic protein motion is in fact strongly coupled to chemical reaction in such a way as to promote catalysis. This result is in concert with both experimental data and interpretations for this and other enzyme systems studied in the laboratories of the two other investigators who have published reviews in this issue. PMID- 12084051 TI - Environmentally coupled hydrogen tunneling. Linking catalysis to dynamics. AB - Many biological C-H activation reactions exhibit nonclassical kinetic isotope effects (KIEs). These nonclassical KIEs are too large (kH/kD > 7) and/or exhibit unusual temperature dependence such that the Arrhenius prefactor KIEs (AH/AD) fall outside of the semiclassical range near unity. The focus of this minireview is to discuss such KIEs within the context of the environmentally coupled hydrogen tunneling model. Full tunneling models of hydrogen transfer assume that protein or solvent fluctuations generate a reactive configuration along the classical, heavy-atom coordinate, from which the hydrogen transfers via nuclear tunneling. Environmentally coupled tunneling also invokes an environmental vibration (gating) that modulates the tunneling barrier, leading to a temperature dependent KIE. These properties directly link enzyme fluctuations to the reaction coordinate for hydrogen transfer, making a quantum view of hydrogen transfer necessarily a dynamic view of catalysis. The environmentally coupled hydrogen tunneling model leads to a range of magnitudes of KIEs, which reflect the tunneling barrier, and a range of AH/AD values, which reflect the extent to which gating modulates hydrogen transfer. Gating is the primary determinant of the temperature dependence of the KIE within this model, providing insight into the importance of this motion in modulating the reaction coordinate. The potential use of variable temperature KIEs as a direct probe of coupling between environmental dynamics and the reaction coordinate is described. The evolution from application of a tunneling correction to a full tunneling model in enzymatic H transfer reactions is discussed in the context of a thermophilic alcohol dehydrogenase and soybean lipoxygenase-1. PMID- 12084052 TI - The evolution of monomeric and oligomeric betagamma-type crystallins. Facts and hypotheses. AB - The case of homologous monomeric gamma-type and oligomeric beta-type crystallins has been described and analyzed in evolutionary terms. Data and hypotheses from molecular genetics and structural investigations converge and suggest a novel three-phase model for the evolutionary history of crystallin-type proteins. In the divergent cascades of monomeric and oligomeric crystallins, a pivotal role was played by alterations in the gene segments encoding the C-terminal extensions and the intermotif or interdomain linker peptides. These were genomic hot spots where evolution experimented to produce the modern variety of betagamma crystallin-type quaternary structures. PMID- 12084053 TI - Unique structural determinants in the signal peptides of "spontaneously" inserting thylakoid membrane proteins. AB - A series of thylakoid membrane proteins, including PsbX, PsbY and PsbW, are synthesized with cleavable signal peptides yet inserted using none of the known Sec/SRP/Tat/Oxa1-type insertion machineries. Here, we show that, although superficially similar to Sec-type signal peptides, these thylakoidal signal peptides contain very different determinants. First, we show that basic residues in the N-terminal domain are not important, ruling out electrostatic interactions as an essential element of the insertion mechanism, and implying a fundamentally different targeting mechanism when compared with the structurally similar M13 procoat. Second, we show that acidic residues in the C-domain are essential for the efficient maturation of the PsbX and PsbY-A1 peptides, and that even a single substitution of the -5 Glu by Val in the PsbX signal peptide abolishes maturation in the thylakoid. Processing efficiency is restored to an extent, but not completely, by the highly hydrophilic Asn, implying that this domain is required to be hydrophilic, but preferably negatively charged, in order to present the cleavage site in an optimal manner. We show that substitution of the PsbX C domain Glu residues by Val leads to a burial of the cleavage site within the bilayer although insertion is unaffected. Finally, we show that substitution of the Glu residues in the lumenal A2 loop of the PsbY polyprotein leads to a block in cleavage on the stromal side of the membrane, and present evidence that the PsbY-A2 signal peptide is required to be relatively hydrophilic and unable to adopt a transmembrane conformation on its own. These data indicate that, rather than being merely additional hydrophobic regions to promote insertion, the signal peptides of these thylakoid proteins are complex domains with uniquely stringent requirements in the C-domain and/or translocated loop regions. PMID- 12084054 TI - Trehalose-based oligosaccharides isolated from the cytoplasm of Mycobacterium smegmatis. Relation to trehalose-based oligosaccharides attached to lipid. AB - A series of trehalose-based oligosaccharides were isolated from the cytoplasmic fraction of Mycobacterium smegmatis and purified by gel-filtration and paper chromatography and TLC. Their structures were determined by HPLC and GLC to determine sugar composition and ratios, MALDI-TOF MS to measure molecular mass, methylation analysis to determine linkages, (1)H-NMR to obtain anomeric configurations of glycosidic linkages, and exoglycosidase digestions followed by TLC to determine sequences and anomeric configurations of the monosaccharides. Six different oligosaccharides were identified all with trehalose as the basic structure and additional glucose or galactose residues attached in various linkages. One of these oligosaccharides is the disaccharide trehalose (Glcalpha1 1alphaGlc), which is present in substantial amounts in these cells and also in other mycobacteria. Two other oligosaccharides, the tetrasaccharides Glcalpha1 4Glcalpha1-1alphaGlc6-1alphaGal and Galalpha1-6Galalpha1-6Glcalpha1-1alphaGlc, have not previously been isolated from natural sources or synthesized chemically. The fourth oligosaccharide, Glcbeta1-6Glcbeta1-6Glcalpha1-1alphaGlc, has been isolated from corynebacteria, but not reported in other organisms. Two other oligosaccharides, Glcalpha1-4Glcalpha1-1alphaGlc, which has been synthesized chemically and isolated from insects but not previously reported in mycobacteria, and Glcbeta1-6Glcalpha1-1alphaGlc, which was previously isolated from Mycobacterium fortuitum and yeast, were also characterized. Another trisaccharide found in the cytosol has been partially characterized as arabinosyl-1-4trehalose, but neither the anomeric configuration nor the D or L configuration of the arabinose is known. In analogy with sucrose and its higher homologs, raffinose and stachyose, which may act as protective agents during maturation drying in plants, these trehalose homologs may also have a protective role in mycobacteria, perhaps during latency. PMID- 12084055 TI - Cloning, chromosomal localization and characterization of the murine mucin gene orthologous to human MUC4. AB - We report here the full coding sequence of a novel mouse putative membrane associated mucin containing three extracellular EGF-like motifs and a mucin-like domain consisting of at least 20 tandem repeats of 124-126 amino acids. Screening a cosmid and a BAC libraries allowed to isolate several genomic clones. Genomic and cDNA sequence comparisons showed that the gene consists of 25 exons and 24 introns covering a genomic region of approximately 52 kb. The first intron is approximately 16 kb in length and is followed by an unusually large exon (approximately 9.5 kb) encoding Ser/Thr-rich tandemly repeated sequences. Radiation hybrid mapping localized this new gene to a mouse region of chromosome 16, which is the orthologous region of human chromosome 3q29 encompassing the large membrane-anchored mucin MUC4. Contigs analysis of the Human Genome Project did not reveal any other mucin on chromosome 3q29 and, interestingly, our analysis allowed the determination of the genomic organization of the human MUC4 and showed that its exon/intron structure is identical to that of the mouse gene we cloned. Furthermore, the human MUC4 shares considerable homologies with the mouse gene. Based on these data, we concluded that we isolated the mouse ortholog of MUC4 we propose as Muc4. Expression studies showed that Muc4 is ubiquitous like SMC and MUC4, with highest levels of expression in trachea and intestinal tract. PMID- 12084056 TI - Monoterpene biosynthesis in lemon (Citrus limon). cDNA isolation and functional analysis of four monoterpene synthases. AB - Citrus limon possesses a high content and large variety of monoterpenoids, especially in the glands of the fruit flavedo. The genes responsible for the production of these monoterpenes have never been isolated. By applying a random sequencing approach to a cDNA library from mRNA isolated from the peel of young developing fruit, four monoterpene synthase cDNAs were isolated that appear to be new members of the previously reported tpsb family. Based on sequence homology and phylogenetic analysis, these sequences cluster in two separate groups. All four cDNAs could be functionally expressed in Escherichia coli after removal of their plastid targeting signals. The main products of the enzymes in assays with geranyl diphosphate as substrate were (+)-limonene (two cDNAs) (-)-beta-pinene and gamma-terpinene. All enzymes exhibited a pH optimum around 7; addition of Mn(2+) as bivalent metal ion cofactor resulted in higher activity than Mg(2+), with an optimum concentration of 0.6 mm. K(m) values ranged from 0.7 to 3.1 microm. The four enzymes account for the production of 10 out of the 17 monoterpene skeletons commonly observed in lemon peel oil, corresponding to more than 90% of the main components present. PMID- 12084058 TI - Role of histidine 42 in ascorbate peroxidase. Kinetic analysis of the H42A and H42E variants. AB - To examine the role of the distal His42 residue in the catalytic mechanism of pea cytosolic ascorbate peroxidase, two site-directed variants were prepared in which His42 was replaced with alanine (H42A) or glutamic acid (H42E). Electronic spectra of the ferric derivatives of H42A and H42E (pH 7.0, mu = 0.10 m, 25.0 degrees C) revealed wavelength maxima [lambda(max) (nm): 397, 509, approximately equal to 540(sh), 644 (H42A); 404, 516, approximately equal to 538(sh), 639 (H42E)] consistent with a predominantly five-co-ordinate high-spin iron. The specific activity of H42E for oxidation of L-ascorbate (8.2 +/- 0.3 U.mg(-1)) was approximately equal to 30-fold lower than that of the recombinant wild-type enzyme (rAPX); the H42A variant was essentially inactive but activity could be partially recovered by addition of exogenous imidazoles. The spectra of the Compound I intermediates of H42A [lambda(max) (nm) = 403, 534, 575(sh), 645] and H42E [lambda(max) (nm) = 404, 530, 573(sh), 654] were similar to those of rAPX. Pre-steady-state data for formation of Compound I for H42A and H42E were consistent with a mechanism involving accumulation of a transient enzyme intermediate (K(d)) followed by conversion of this intermediate into Compound I (k'(1)). Values for k'(1) and K(d) were, respectively, 4.3 +/- 0.2 s(-1) and 30 +/- 2.0 mM (H42A) and 28 +/- 1.0 s(-1) and 0.09 +/- 0.01 mM (H42E). Photodiode array experiments for H42A revealed wavelength maxima for this intermediate at 401 nm, 522 nm and 643 nm, consistent with the formation of a transient [H42A H(2)O(2)] species. Rate constants for Compound I formation for H42A were independent of pH, but for rAPX and H42E were pH-dependent [pKa = 4.9 +/- 0.1 (rAPX) and pK(a) = 6.7 +/- 0.2 (H42E)]. The results provide: (a) evidence that His42 is critical for Compound I formation in APX; (b) confirmation that titration of His42 controls Compound I formation and an assignment of the pK(a) for this group; (c) mechanistic and spectroscopic evidence for an intermediate before Compound I formation; (d) evidence that a glutamic acid residue at position 42 can act as the acid-base catalyst in ascorbate peroxidase. PMID- 12084057 TI - Functional integration of mitochondrial and hydrogenosomal ADP/ATP carriers in the Escherichia coli membrane reveals different biochemical characteristics for plants, mammals and anaerobic chytrids. AB - The expression of mitochondrial and hydrogenosomal ADP/ATP carriers (AACs) from plants, rat and the anaerobic chytridiomycete fungus Neocallimastix spec. L2 in Escherichia coli allows a functional integration of the recombinant proteins into the bacterial cytoplasmic membrane. For AAC1 and AAC2 from rat, apparent Km values of about 40 microm for ADP, and 105 microm or 140 microm, respectively, for ATP have been determined, similar to the data reported for isolated rat mitochondria. The apparent Km for ATP decreased up to 10-fold in the presence of the protonophore m-chlorocarbonylcyanide phenylhydrazone (CCCP). The hydrogenosomal AAC isolated from the chytrid fungus Neocallimastix spec. L2 exhibited the same characteristics, but the affinities for ADP (165 microm) and ATP (2.33 mm) were significantly lower. Notably, AAC1-3 from Arabidopsis thaliana and AAC1 from Solanum tuberosum (potato) showed significantly higher external affinities for both nucleotides (10-22 microm); they were only slightly influenced by CCCP. Studies on intact plant mitochondria confirmed these observations. Back exchange experiments with preloaded E. coli cells expressing AACs indicate a preferential export of ATP for all AACs tested. This is the first report of a functional integration of proteins belonging to the mitochondrial carrier family (MCF) into a bacterial cytoplasmic membrane. The technique described here provides a relatively simple and highly reproducible method for functional studies of individual mitochondrial-type carrier proteins from organisms that do not allow the application of sophisticated genetic techniques. PMID- 12084059 TI - A Ca2+/CaM-dependent kinase from pea is stress regulated and in vitro phosphorylates a protein that binds to AtCaM5 promoter. AB - An immuno-homologue of maize Ca2+/calmodulin (CaM)-dependent protein kinase with a molecular mass of 72 kDa was identified in pea. The pea kinase (PsCCaMK) was upregulated in roots in response to low temperature and increased salinity. Exogenous Ca2+ application increased the kinase level and the response was faster than that obtained following stress application. Low temperature-mediated, but not salinity-mediated stress kinase increase was inhibited by the application of EGTA and W7, a CaM inhibitor. The purification of PsCCaMK using immuno-affinity chromatography resulted in coelution of the kinase with another polypeptide of molecular mass 40 kDa (p40). Western blot revealed the presence of PsCCaMK in nuclear protein extracts and was found to phosphorylate p40 in vitro. Gel mobility shift and South-Western analysis showed that p40 is a DNA-binding protein and it interacted specifically with one of the cis acting elements of the Arabidopsis CaM5 gene (AtCaM5) promoter. The binding of p40 to the specific elements in the AtCaM5 promoter was dependent of its dephosphorylated state. Our results suggest that p40 could be an upstream signal component of the stress responses. PMID- 12084060 TI - Balanced expression of single subunits in a multisubunit protein, achieved by cell fusion of individual transfectants. AB - To establish stable cell lines that produce recombinant multisubunit proteins, it is usually necessary to cotransfect cells with several independent gene constructs. Here, we show that a stepwise fusion of individually transfected cells, results in a fused cell-line that secretes a complete multisubunit protein. Functional expression of recombinant multisubunit proteins may require a defined expression ratio between each protein subunit. The cell-fusion technology described allows a predefined expression level of each subunit. Using SIgA as a model protein we demonstrate that the majority of the fused cells inherit the molar expression ratio of the parental transfected cells. These results indicate that simplified screening of clones expressing the expected subunit ratios may be possible using the cell-fusion technology. This technology may therefore be an alternative to generic transfection methods for the establishment of cells that produce multiprotein complexes such as antibodies, receptors, ion channels and other multisubunit proteins. PMID- 12084061 TI - Transglutaminase-mediated polyamination of vasoactive intestinal peptide (VIP) Gln16 residue modulates VIP/PACAP receptor activity. AB - Previous data showing an increase of receptor binding activity of [R16]VIP, a vasoactive intestinal peptide (VIP) structural analogue containing arginine at the position 16 of its amino acid sequence, have pointed out the importance of a positive charge at this site. Here, the functional characterization of three VIP polyaminated adducts (VIPDap, VIPSpd, and VIPSpm), obtained by a transglutaminase catalysed reaction between the VIP Gln16 residue and 1,3-diaminopropane (Dap), spermidine (Spd), or spermine (Spm), is reported. Appropriate binding assays and adenylate cyclase enzymatic determinations have shown that these VIP adducts act as structural VIP agonists, both in vitro and in vivo. In particular, their IC50 and EC50 values of human and rat VIP/pituitary adenylate cyclase activating peptide (PACAP)1 and VIP/PACAP2 receptors indicate that VIPDap is a VIP agonist, with an affinity and a potency higher than that of VIP, while VIPSpd and VIPSpm are also agonists but with affinities lower than that of VIP. These findings suggest that the difference in adduct agonist activity reflects the differences in the positive charge and carbon chain length of the polyamine covalently linked with the VIP Gln16 residue. In addition, the data obtained strongly suggest that the length of polyamine carbon chain could be critical for the interaction of the agonist with its receptor, even though possible hydrophobic interaction cannot be ruled out. In vivo experiments on murine J774 macrophage cell cultures have shown the ability of these compounds to stimulate the inducible nitric oxide synthase activity at the transcriptional level. PMID- 12084062 TI - Amphipathic property of free thiol group contributes to an increase in the catalytic efficiency of carboxypeptidase Y. AB - Cys341 of carboxypeptidase Y, which constitutes one side of the solvent accessible surface of the S1 binding pocket, was replaced with Gly, Ser, Asp, Val, Phe or His by site-directed mutagenesis. Kinetic analysis, using Cbz dipeptide substrates, revealed that polar amino acids at the 341 position increased K(m) whereas hydrophobic amino acids in this position tended to decrease K(m). This suggests the involvement of Cys341 in the formation of the Michaelis complex in which Cys341 favors the formation of hydrophobic interactions with the P1 side chain of the substrate as well as with residues comprising the surface of the S1 binding pocket. Furthermore, C341G and C341S mutants had significantly higher k(cat) values with substrates containing the hydrophobic P1 side chain than C341V or C341F. This indicates that the nonhydrophobic property conferred by Gly or Ser gives flexibility or instability to the S1 pocket, which contributes to the increased k(cat) values of C341G or C341S. The results suggest that Cys341 may interact with His397 during catalysis. Therefore, we propose a dual role for Cys341: (a) its hydrophobicity allows it to participate in the formation of the Michaelis complex with hydrophobic substrates, where it maintains an unfavorable steric constraint in the S1 subsite; (b) its interaction with the imidazole ring of His397 contributes to the rate enhancement by stabilizing the tetrahedral intermediate in the transition state. PMID- 12084063 TI - Synthesis of phosphoenol pyruvate (PEP) analogues and evaluation as inhibitors of PEP-utilizing enzymes. AB - The synthesis of 10 new phosphoenolpyruvate (PEP) analogues with modifications in the phosphate and the carboxylate function is described. Included are two potential irreversible inhibitors of PEP-utilizing enzymes. One incorporates a reactive chloromethylphosphonate function replacing the phosphate group of PEP. The second contains a chloromethyl group substituting for the carboxylate function of PEP. An improved procedure for the preparation of the known (Z)- and (E)-3-chloro-PEP is also given. The isomers were obtained as a 4 : 1 mixture, resolved by anion-exchange chromatography after the last reaction step. The stereochemistry of the two isomers was unequivocally assigned from the (3)J(H-C) coupling constants between the carboxylate carbons and the vinyl protons. All of these and other known PEP-analogues were tested as reversible and irreversible inhibitors of Mg2+- and Mn2+- activated PEP-utilizing enzymes: enzyme I of the phosphoenolpyruvate:sugar phosphotransferase system (PTS), pyruvate kinase, PEP carboxylase and enolase. Without exception, the most potent inhibitors were those with substitution of a vinyl proton. Modification of the phosphate and the carboxylate groups resulted in less effective compounds. Enzyme I was the least tolerant to such modifications. Among the carboxylate-modified analogues, only those replaced by a negatively charged group inhibited pyruvate kinase and enolase. Remarkably, the activity of PEP carboxylase was stimulated by derivatives with neutral groups at this position in the presence of Mg2+, but not with Mn2+. For the irreversible inhibition of these enzymes, (Z)-3-Cl-PEP was found to be a very fast-acting and efficient suicide inhibitor of enzyme I (t(1/2) = 0.7 min). PMID- 12084064 TI - Ligand-induced heterodimerization between the ligand binding domains of the Drosophila ecdysteroid receptor and ultraspiracle. AB - The insect ecdysteroid receptor consists of a heterodimer between EcR and the RXR orthologue, USP. We addressed the question of whether this heterodimer, like all other RXR heterodimers, may be formed in the absence of ligand and whether ligand promotes dimerization. We found that C-terminal protein fragments that comprised the ligand binding, but not the DNA binding domain of EcR and USP and which were equipped with the activation or DNA binding region of GAL4, respectively, exhibit a weak ability to interact spontaneously with each other. Moreover, the heterodimer formation is greatly enhanced upon administration of active ecdysteroids in a dose-dependent manner. This was shown in vivo by a yeast two hybrid system and in vitro by a modified electromobility shift assay. Furthermore, the EcR fragment expressed in yeast was functional and bound radioactively labelled ecdysteroid specifically. Ligand binding was greatly enhanced by the presence of a USP ligand binding domain. Therefore, ecdysteroids are capable of inducing heterodimer formation between EcR and USP, even when the binding of these receptor proteins to cognate DNA response elements does not occur. This capability may be a regulated aspect of ecdysteroid action during insect development. PMID- 12084065 TI - The S100A8/A9 protein as a partner for the cytosolic factors of NADPH oxidase activation in neutrophils. AB - In a previous study, the S100A8/A9 protein, a Ca2+- and arachidonic acid-binding protein, abundant in neutrophil cytosol, was found to potentiate the activation of the redox component of the O2- generating oxidase in neutrophils, namely the membrane-bound flavocytochrome b, by the cytosolic phox proteins p67phox, p47phox and Rac (Doussiere J., Bouzidi F. and Vignais P.V. (2001) Biochem. Biophys. Res. Commun.285, 1317-1320). This led us to check by immunoprecipitation and protein fractionation whether the cytosolic phox proteins could bind to S100A8/A9. Following incubation of a cytosolic extract from nonactivated bovine neutrophil with protein A-Sepharose bound to anti-p67phox antibodies, the recovered immunoprecipitate contained the S100 protein, p47phox and p67phox. Cytosolic protein fractionation comprised two successive chromatographic steps on hydroxyapatite and DEAE cellulose, followed by isoelectric focusing. The S100A8/A9 heterodimeric protein comigrated with the cytosolic phox proteins, and more particularly with p67phox and Rac2, whereas the isolated S100A8 protein displayed a tendancy to bind to p47phox. Using a semirecombinant cell-free system of oxidase activation consisting of recombinant p67phox, p47phox and Rac2, neutrophil membranes and arachidonic acid, we found that the S100A8/A9-dependent increase in the elicited oxidase activity corresponded to an increase in the turnover of the membrane-bound flavocytochrome b, but not to a change of affinity for NADPH or O2. In the absence of S100A8/A9, oxidase activation departed from michaelian kinetics above a critical threshold concentration of cytosolic phox proteins. Addition of S100A8/A9 to the cell-free system rendered the kinetics fully michaelian. The propensity of S100A8/A9 to bind the cytosolic phox proteins, and the effects of S100A8/A9 on the kinetics of oxidase activation, suggest that S100A8/A9 might be a scaffold protein for the cytosolic phox proteins or might help to deliver arachidonic acid to the oxidase, thus favoring the productive interaction of the cytosolic phox proteins with the membrane-bound flavocytochrome b. PMID- 12084066 TI - Pyruvate decarboxylase from Kluyveromyces lactis. An enzyme with an extraordinary substrate activation behaviour. AB - Pyruvate decarboxylase (EC 4.1.1.1) was isolated and purified from the yeast Kluyveromyces lactis. The properties of this enzyme relating to the native oligomeric state, the subunit size, the nucleotide sequence of the coding gene(s), the catalytic activity, and protein fluorescence as well as circular dichroism are very similar to those of the well characterized pyruvate decarboxylase species from yeast. Remarkable differences were found in the substrate activation behaviour of the two pyruvate decarboxylases using three independent methods: steady-state kinetics, stopped-flow measurements, and kinetic dilution experiments. The dependence of the observed activation rate constant on the substrate concentration of pyruvate decarboxylase from K. lactis showed a minimum at a pyruvate concentration of 1.5 mm. According to the mechanism of substrate activation suggested this local minimum occurs due to the big ratio of the dissociation constants for the binding of the first (regulatory) and the second (catalytic) substrate molecule. The microscopic rate constants of the substrate activation could be determined by a refined fit procedure. The influence of the artificial activator pyruvamide on the activation of the enzyme was studied. PMID- 12084067 TI - Structure of human immunodeficiency virus type 1 Vpr(34-51) peptide in micelle containing aqueous solution. AB - Human immunodeficiency virus type 1 protein R (HIV-1 Vpr) promotes nuclear entry of viral nucleic acids in nondividing cells, causes G(2) cell cycle arrest and is involved in cellular differentiation and cell death. Vpr subcellular localization is as variable as its functions. It is known, that consistent with its role in nuclear transport, Vpr localizes to the nuclear envelope of human cells. Further, a reported ion channel activity of Vpr is clearly dependent on its localization in or at membranes. We focused our structural studies on the secondary structure of a peptide consisting of residues 34-51 of HIV-1 Vpr. This part of Vpr plays an important role in Vpr oligomerization, contributes to cell cycle arrest activity, and is essential for virion incorporation and binding to HHR23A, a protein involved in DNA repair. Employing NMR spectroscopy we found this part of Vpr to be almost completely alpha helical in the presence of micelles, as well as in trifluoroethanol containing and methanol/chloroform solvent. Our results provide structural data suggesting residues 34-51 of Vpr to contain an amphipathic, leucine-zipper-like alpha helix, which serves as a basis for oligomerization of Vpr and its interactions with cellular and viral factors involved in subcellular localization and virion incorporation of Vpr. PMID- 12084068 TI - Synthesis, characterization and application of two nucleoside triphosphate analogues, GTPgammaNH(2) and GTPgammaF. AB - Guanosine triphosphate nucleotide analogues such as GppNHp (also named GMPPNP) or GTPgammaS are widely used to stabilize rapidly hydrolyzing protein-nucleotide complexes and to investigate biochemical reaction pathways. Here we describe the chemical synthesis of guanosine 5'-O-(gamma-amidotriphosphate) (GTPgammaNH(2)) and a new synthesis of guanosine 5'-O-(gamma-fluorotriphosphate) (GTPgammaF). The two nucleotides were characterized using NMR spectroscopy and isothermal titration calorimetry. Chemical shift data on (31)P, (19)F and (1)H NMR resonances are tabulated. For GTPgammaNH(2) the enthalpy of magnesium coordination is DeltaH degrees = 3.9 kcal.mol(-1) and the association constant K(a) is 0.82 mm(-1). The activation energy for GTPgammaNH(2).Mg2+ complex formation is DeltaH++ = 7.8 +/- 0.15 kcal.mol(-1), similar to that for the natural substrate GTP. For GTPgammaF we obtained a similar enthalpy of DeltaH degrees = 3.9 kcal.mol(-1) while the magnesium association constant is only K(a) = 0.2 mm(-1). The application of both guanine nucleotide analogues to the GTP binding protein Ras was investigated. The rate of hydrolysis of GTPgammaNH(2) bound to Ras protein lay between the rates found for Ras-bound GTPgammaS and GppNHp, while Ras-catalysed hydrolysis of GTPgammaF was almost as fast as for GTP. The two compounds extend the variety of nucleotide analogues and may prove useful in structural, kinetic and cellular studies. PMID- 12084069 TI - Shb links SLP-76 and Vav with the CD3 complex in Jurkat T cells. AB - This study addresses the interactions between the adaptor protein Shb and components involved in T cell signalling, including SLP-76, Gads, Vav and ZAP70. We show that both SLP-76 and ZAP70 co-immunoprecipitate with Shb in Jurkat T cells and that Shb and Vav co-immunoprecipitate when cotransfected in COS cells. We also demonstrate, utilizing fusion protein constructs, that SLP-76, Gads and Vav associate independently of each other to different domains or regions, of Shb. Overexpression of an SH2 domain-defective Shb causes diminished phosphorylation of SLP-76 and Vav and consequently decreased activation of c-Jun kinase upon T cell receptor (TCR) stimulation. Shb was also found to localize to glycolipid-enriched membrane microdomains (GEMs), also called lipid rafts, after TCR stimulation. Our results indicate that upon TCR stimulation, Shb is targeted to these lipid rafts where Shb aids in recruiting the SLP-76-Gads-Vav complex to the T cell receptor zeta-chain and ZAP70. PMID- 12084070 TI - The structure of the O-chain of the lipopolysaccharide of a prototypal diarrheagenic strain of Hafnia alvei that has characteristics of a new species under the genus Escherichia. AB - The structure of the O-polysaccharide of the lipopolysaccharide from a diarrheal strain isolated in Bangladesh was studied with sugar, and methylation analysis, NMR spectroscopy, mass spectrometry and partial acid hydrolysis. The strain was first designated as Hafnia alvei, but later found to be a possible new species in the genus Escherichia. Two different polysaccharides were detected, a major and a minor one. The structure of the major polysaccharide is given below, while the structure of the minor one was not investigated. The structure of the repeating unit was established as The structure does not resemble any of the previously investigated lipopolysaccharide O-chains from Escherichia coli or H. alvei, but could fit in either group based on types of sugar residues and acidity. Phenotypic microbiological studies cannot definitely assign it to either species of the two genera. Genetic hybridization studies indicate that the Bangladeshi isolates may require a new species designation under the genus Escherichia. PMID- 12084071 TI - Herbaspirillum seropedicae signal transduction protein PII is structurally similar to the enteric GlnK. AB - PII-like proteins are signal transduction proteins found in bacteria, archaea and eukaryotes. They mediate a variety of cellular responses. A second PII-like protein, called GlnK, has been found in several organisms. In the diazotroph Herbaspirillum seropedicae, PII protein is involved in sensing nitrogen levels and controlling nitrogen fixation genes. In this work, the crystal structure of the unliganded H. seropedicae PII was solved by X-ray diffraction. H. seropedicae PII has a Gly residue, Gly108 preceding Pro109 and the main-chain forms a beta turn. The glycine at position 108 allows a bend in the C-terminal main-chain, thereby modifying the surface of the cleft between monomers and potentially changing function. The structure suggests that the C-terminal region of PII proteins may be involved in specificity of function, and nonenteric diazotrophs are found to have the C-terminal consensus XGXDAX(107-112). We are also proposing binding sites for ATP and 2-oxoglutarate based on the structural alignment of PII with PII-ATP/GlnK-ATP, 5-carboxymethyl-2-hydroxymuconate isomerase and 4 oxalocrotonate tautomerase bound to the inhibitor 2-oxo-3-pentynoate. PMID- 12084072 TI - Changes in ultrastructure and the occurrence of permeability transition in mitochondria during rat liver regeneration. AB - Mitochondrial bioenergetic impairment has been found in the organelles isolated from rat liver during the prereplicative phase of liver regeneration. To gain insight into the mechanism underlying this impairment, we investigated mitochondrial ultrastructure and membrane permeability properties in the course of liver regeneration after partial hepatectomy, with special interest to the role played by Ca2+ in this process. The results show that during the first day after partial hepatectomy, significant changes in the ultrastructure of mitochondria in situ occur. Mitochondrial swelling and release from mitochondria of both glutamate dehydrogenase and aspartate aminotransferase isoenzymes with an increase in the mitochondrial Ca2+ content were also observed. Cyclosporin-A proved to be able to prevent the changes in mitochondrial membrane permeability properties. At 24 h after partial hepatectomy, despite alteration in mitochondrial membrane permeability properties, no release of cytochrome c was found. The ultrastructure of mitochondria, the membrane permeability properties and the Ca2+ content returned to normal values during the replicative phase of liver regeneration. These results suggest that, during the prereplicative phase of liver regeneration, the changes in mitochondrial ultrastructure observed in liver specimens were correlated with Ca2+-induced permeability transition in mitochondria. PMID- 12084073 TI - Expression of the aspartate/glutamate mitochondrial carriers aralar1 and citrin during development and in adult rat tissues. AB - Aralar1 and citrin are members of the subfamily of calcium-binding mitochondrial carriers and correspond to two isoforms of the mitochondrial aspartate/glutamate carrier (AGC). These proteins are activated by Ca2+ acting on the external side of the inner mitochondrial membrane. Although it is known that aralar1 is expressed mainly in skeletal muscle, heart and brain, whereas citrin is present in liver, kidney and heart, the precise tissue distribution of the two proteins in embryonic and adult tissues is largely unknown. We investigated the pattern of expression of aralar1 and citrin in murine embryonic and adult tissues at the mRNA and protein levels. In situ hybridization analysis indicates that both isoforms are expressed strongly in the branchial arches, dermomyotome, limb and tail buds at early embryonic stages. However, citrin was more abundant in the ectodermal components of these structures whereas aralarl had a predominantly mesenchymal localization. The strong expression of citrin in the liver was acquired postnatally, whereas the characteristic expression of aralar1 in skeletal muscle was detected at E18 and that in the heart began early in development (E11) and was preferentially localized to auricular myocardium in late embryonic stages. Aralar1 was also expressed in bone marrow, T-lymphocytes and macrophages, including Kupffer cells in the liver, indicating that this is the major AGC isoform present in the hematopoietic system. Both aralar1 and citrin were expressed in fetal gut and adult stomach, ovary, testis, and pancreas, but only aralar1 is enriched in lung and insulin-secreting beta cells. These results show that aralar1 is expressed in many more tissues than originally believed and is absent from hepatocytes, where citrin is the only AGC isoform present. This explains why citrin deficiency in humans (type II citrullinemia) only affects the liver and suggests that aralar1 may compensate for the lack of citrin in other tissues. PMID- 12084074 TI - The cold-active lipase of Pseudomonas fragi. Heterologous expression, biochemical characterization and molecular modeling. AB - A recombinant lipase cloned from Pseudomonas fragi strain IFO 3458 (PFL) was found to retain significant activity at low temperature. In an attempt to elucidate the structural basis of this behaviour, a model of its three dimensional structure was built by homology and compared with homologous mesophilic lipases, i.e. the Pseudomonas aeruginosa lipase (45% sequence identity) and Burkholderia cepacia lipase (38%). In this model, features common to all known lipases have been identified, such as the catalytic triad (S83, D238 and H260) and the oxyanion hole (L17, Q84). Structural modifications recurrent in cold-adaptation, i.e. a large amount of charged residues exposed at the protein surface, have been detected. Noteworthy is the lack of a disulphide bridge conserved in homologous Pseudomonas lipases that may contribute to increased conformational flexibility of the cold-active enzyme. PMID- 12084075 TI - The plastid transcription kinase from mustard (Sinapis alba L.). A nuclear encoded CK2-type chloroplast enzyme with redox-sensitive function. AB - The plastid transcription kinase (PTK), a component of the major RNA polymerase complex from mustard chloroplasts, has been implicated in redox-mediated regulation of plastid gene expression. A cloning strategy to define the PTK gene(s) resulted in the isolation of a full-length cDNA for a protein with overall high homology with the alpha subunit of cytosolic casein kinase (CK2) that contained an N-terminal extension for a putative plastid transit peptide. Using in organello chloroplast import studies, immunodetection and MS, we found that the corresponding protein, termed cpCK2alpha, is targeted to the chloroplast and is associated with the plastid RNA polymerase PEP-A. The bacterially overexpressed protein shows CK2 kinase activity and is subject to glutathione inhibition in the same way as authentic chloroplast PTK. Furthermore, it readily phosphorylates components of the plastid transcription apparatus in vitro with a substrate specificity similar to that of PTK. PMID- 12084076 TI - Positive patch tests with a dermatophagoides mix relate to an increased responsiveness to standard patch test allergens. AB - The diagnostic meaningfulness of patch tests with house dust mite allergens is still questionable. Our own impression has been that positive results with a dermatophagoides mix may occur preferentially in patients with a generally enhanced responsiveness to contact allergens. To check this, all of our patients allocated to patch testing with the standard series were additionally patch tested with a dermatophagoides mix by the same technique that was used for standard contact allergens. Out of 571 patients tested, 188 showed delayed responses to this mix that were indistinguishable from typical allergic patch test reactions but of no apparent clinical relevance. No relationship was found between positive dermatophagoides patch tests and an atopic disposition of the patients or characteristics of their eczema. However, 64.4% of the patients with a positive dermatophagoides patch test showed a response to at least 1 contact allergen of the standard series, compared to only 56.4% of the patients without a positive dermatophagoides reaction (p < 0.05). The reactivity to the mite mix was not related to the responsiveness towards any particular contact allergens. We suppose that some unidentified factors may contribute to positive reactions to the dermatophagoides mix that may also favour an enhanced general responsiveness to contact allergens. PMID- 12084077 TI - Patch tests with thiurams at 0.25% pet. and 1% pet. are of equal diagnostic value. AB - Thiuram mix is tested in the standard series at a test concentration of 1% pet. The single thiurams (DPTD, TMTD, TMTM, TETD), however, are usually tested at 0.25% pet. in Germany. In other countries, the individual components of thiuram mix are tested at 1% pet. The German Contact Dermatitis Research Group (DKG) compared both patch test concentrations in 530 patients in order to find out if (i) a significant number of positive patch tests are missed by testing at the lower concentration, (ii) problems with irritant test reactions occur by increasing the test concentration to 1%, and (iii) the sensitivity of the thiuram mix rises when the breakdown test is done with the higher concentration. Slightly more positive reactions were seen with the higher concentration, but this increase did not reach statistical significance. The reaction index, as a measure for the relation of positive to irritant and/or questionable reactions, remained unchanged for the individual thiurams. The sensitivity of the mix also did not change when the breakdown test was performed with 1% pet. instead of 0.25% pet. Thus, we conclude that both concentrations are of equal diagnostic value in patch testing. PMID- 12084078 TI - Contact urticaria due to polyethylene gloves. AB - We report a rare case of contact urticaria due to polyethylene gloves. The patient, a 46-year-old cook, had had had chronic urticaria since 1985, and first visited our hospital in June 2000. We began by prescribing antihistamine and antiallergenic drugs for him, but his condition did not improve. From a detailed interview, we established that when he put on polyethylene gloves at work, his condition worsened. We suspected some component of his gloves to be the cause of his symptoms. Prick and scratch tests with a solution extracted from his gloves showed a wheal-and-flare reaction at 15 min. We advised him to wear a cotton shirt under his clothes in daily life, and to put on cotton gloves under his polyethylene gloves while at work. Subsequently, the size and the number of wheals were markedly smaller and the subject's symptoms were reduced. PMID- 12084079 TI - Studies on the autoxidation and sensitizing capacity of the fragrance chemical linalool, identifying a linalool hydroperoxide. AB - Fragrances are among the most common causes of allergic contact dermatitis. The two monoterpenes linalool and d-limonene are the most frequently incorporated fragrance chemicals in scented products. Previous studies on d-limonene show that this monoterpene oxidizes on air exposure (autoxidation) and that allergenic oxidation products are formed. Due to structural similarities, linalool might also form allergenic oxidation products on air exposure. The aim of the present study was to study the autoxidation of linalool and to investigate the sensitizing potential of linalool before and after air exposure. Linalool was oxidized for 10 weeks and gas chromatographic analyses showed that the content of linalool decreased to about 80%. The chromatograms revealed the formation of other compounds during oxidation. One of the major oxidation products was isolated and identified as 7-hydroperoxy-3,7-dimethyl-octa-1,5-diene-3-ol. This substance is, to the best of our knowledge, described for the first time. In sensitization studies in guinea pigs, linalool of high purity gave no reactions, while linalool that had been oxidized for 10 weeks sensitized the animals. It is concluded that autoxidation of linalool is essential for its sensitizing potential. PMID- 12084080 TI - Prognosis and work absence due to occupational contact dermatitis. AB - The consequences of a diagnosis of occupational contact dermatitis (OCD) were investigated using cases available from the voluntary surveillance scheme, EPIDERM. Cases of OCD reported from November 1994 to September 1995 were identified and sampled to give at least 100 cases of allergic, irritant and mixed OCD reported by consultant dermatologists (344 cases) and occupational physicians (377 cases). A questionnaire was sent to the reporting physician to elicit further information. 512 completed questionnaires were returned, of which 510 were eligible for analysis. Among cases reported by dermatologists (n = 286) and occupational physicians (n = 224), 7% (6.3%) had been unemployed and 16.8% (20.1%) had taken sick leave. 3 factors independently predicted time off work in a logistic regression analysis: age OR = 1.25 (95% CI, 1.05-1.49), allergic dermatitis OR = 1.77 (95% CI, 1.13-2.79) and medicolegal assessment OR = 4.42 (95% CI, 2.20-8.89). Overall, 15.7% did not improve clinically between the first and last visit. Those who did not improve had been exposed to the agent for longer (mean 7.6 years) than those who did (5.3 years) (p = 0.09). In patients gemifloxacin (MIC50, 0.5 mg/L) > levofloxacin = gatifloxacin (MIC50, 1 mg/L) > BMS284756 (MIC50, 4 mg/L). The MIC50 value for BMS284756 versus Stenotrophomonas/Burkholderia group was 2 mg/L. This potency was similar to that of other quinolones reported previously. The MIC50 results for the Acinetobacter spp. were ciprofloxacin at >2 mg/L, levofloxacin, gatifloxacin and gemifloxacin at 4 mg/L, and BMS284756 at >4 mg/L, all intermediate or resistant concentrations. The E test results compared favorably with the reference dilution test results for P. aeruginosa, Stenotrophomonas/Burkholderia group, and Acinetobacter spp., with an overall essential agreement of 97.0% +/- 2 log2 dilution steps. E test MIC results tended to be slightly lower when testing Stenotrophomonas/Burkholderia group strains. The disk diffusion method correlated well for P. aeruginosa (r = 0.94), Stenotrophomonas/Burkholderia group (r = 0.84) and Acinetobacter spp. (r = 0.99) strains. CONCLUSIONS: BMS284756 was generally less active than other comparison quinolones when tested against non-fermentative Gram-negative bacilli. Its spectrum remains equivalent only if dosing schedules substantiate a proposed susceptible breakpoint of < or =4 mg/L. PMID- 12084102 TI - Lack of correlation between Bartonella DNA detection within fleas, serological results, and results of blood culture in a Bartonella-infected stray cat population. AB - OBJECTIVE: To correlate the presence of different Bartonella species in the blood of a stray cat population trapped on a French military base with specific antibodies and species detected in cat fleas. METHODS: The prevalence of Bartonella bacteremia was investigated in 61 cats by plating frozen whole blood on blood agar plates. Identification of isolates and detection of Bartonella DNA from cat flea batches from ten cats was achieved by PCR amplification and sequencing. Antibody detection was performed by microimmunofluorescence. RESULTS: We obtained 38 isolates of Bartonella from blood. Sixteen were identified as B. clarridgeiae, 15 as B. henselae genotype/serotype Houston 1 (type I), and seven as B. henselae genotype/serotype Marseille (type II). B. henselae was detected in five fleas, and B. clarridgeiae in one flea. Sixty-one per cent of the cats had detectable antibodies against at least one species or serotype. Sixteen cats had antibodies against only one antigen. For each species, the distribution of bacteremia among the cats could not be correlated with either the distribution of infected fleas or the distribution of specific antibodies. CONCLUSIONS: The lack of correlation between Bartonella DNA detection within fleas, serological results, and results of blood culture is probably due to a lack of natural heterologous protection between species or serotypes. Cats suffer bacteremia with three Bartonella species and should therefore be considered the reservoirs of at least three human pathogens. PMID- 12084103 TI - Detection of GB virus C/hepatitis G virus in semen and saliva of HIV type-1 infected men. AB - OBJECTIVE: To investigate the presence of the genome of GB virus C/hepatitis G virus (GBV-C/HGV) in semen and saliva from HIV-1-infected men. METHODS: Samples of blood from 33 men seropositive for HIV-1 were tested for the presence of GBV C/HGV markers of infection, RNA by RT-PCR, and anti-E2 antibodies by ELISA, respectively. The cell-free fractions of seminal fluid and saliva samples of the patients with positive blood samples for GBV-C/HGV RNA or anti-E2 antibodies were then analyzed for the presence of the RNA of this virus. In addition, six semen samples and 11 saliva samples from GBV-C/HGV-negative men were tested. RESULTS: The GBV-C/HGV RNA tested by RT-PCR was recovered from blood in 11 patients of 33 (33.3%), and the antibodies to E2 envelope protein were detected in six patients (18.2%). Since no patient was positive for both markers, the overall prevalence of GBV-C/HGV infection was 51.5% in the studied population. Four-all belonging to the homosexual risk group-of the 17 men with markers to GBV-C/HGV in blood were found to be positive for GBV-C/HGV RNA in mucosal samples: two of them exhibited genomic RNA in both semen and saliva, and two others were positive for semen only. The absence of inhibitors of the PCR technique was confirmed in all mucosal fractions found negative for GBV-C/HGV RNA, except for one saliva sample and one seminal fluid sample. CONCLUSION: These results confirm the high prevalence of GBV-C/HGV infection in patients infected with HIV-1 by sexual exposure and the presence of GBV-C/HGV RNA in seminal fluid and saliva of men with markers of this virus in the blood, suggesting that mucosal fluids could be a potential source for the spread of the GBV-C/HGV infection. PMID- 12084104 TI - Seroprevalence of chronic Chlamydia pneumoniae infection in patients affected by chronic stable asthma. AB - OBJECTIVE: To evaluate the seroprevalence of Chlamydia pneumoniae and age, gender and smoking habits in stable asthmatic patients. METHODS: Over a period of 3 months, 197 adult patients affected by intermittent-to-severe chronic asthma were enrolled from 16 respiratory disease units in the south of Italy. As a control group, we tested 185 healthy, non-asthmatic subjects matched for age and gender, recruited among hospital staff. All patients were submitted to clinical examination, spirometry and blood collection for C. pneumoniae serology. The presence of infection was investigated by microimmunofluorescence (Micro-IF Test) for C. pneumoniae-specific IgG, IgM and IgA antibodies. RESULTS: C. pneumoniae IgG titers > or =1 : 64 were detected in 30.4% of asthmatics and in 30.8% of controls. Correlation of age, gender and smoking habit with C. pneumoniae seropositivity was evaluated by linear regression analysis. Age was significantly associated with C. pneumoniae IgG titer > or =1 : 64 when seropositive asthmatics were tested. Moreover, C. pneumoniae seroprevalence was higher among smokers with a diagnosis of chronic asthma. CONCLUSIONS: The seroprevalence of C. pneumoniae in stable asthmatics was comparable with the controls; therefore, the study does not support the association between C. pneumoniae antibody titers and stable asthma. However, the analysis for likely confounders such as age, gender and smoking status suggests a possible association of enhanced susceptibility to C. pneumoniae infection with age and smoking habitus. PMID- 12084105 TI - Transfusion-transmitted virus prevalence in subjects at high risk of sexually transmitted infection in Turkey. AB - OBJECTIVE: To assess the possible sexual transmission of virus and to identify the prevalence of TTV viremia in Turkey and its association with other hepatotropic viruses. METHODS: Serum samples were collected from 81 subjects (74 prostitutes and seven homosexual men) at high risk of sexually transmitted infection and from 81 healthy controls (74 females and seven males). Sera of patients and controls were tested for TTV, hepatitis A virus, hepatitis B virus, hepatitis C virus, and human immunodeficiency virus. Also, serum alanine and aspartate aminotransferases were measured. RESULTS: The prevalence rates of TTV viremia in the risk group and control group were 86.4% and 82.7%, respectively. There was a statistical difference in mean age between TTV-infected and uninfected subjects (38.6 +/- 9.9 versus 32.2 +/- 6.1 years, respectively, P < 0.001). Prevalence rates of TTV infection in subjects with positive anti-HAV and positive anti-HBc were high when compared with subjects who were negative for these. CONCLUSION: We suggest that TTV infection has a diverse route of transmission, and its prevalence increases with age; also, the prevalence rate of TTV is high in certain risk groups. The prevalence rates of TTV in the group at risk for sexual transmission (86.4%) and in the control group (82.7%) were among the highest ever reported in the world. Also, we suggest that TTV generally does not cause clinical disease, in spite of this high prevalence. PMID- 12084106 TI - Comparative activity of linezolid against staphylococci and enterococci isolated in Italy. AB - The activity of linezolid, a new oxazolidinone, was tested against 862 Gram positive cocci isolated in Italy and compared with the activities of 12 antibiotics. Overall, MIC90s for linezolid (2-4 mg/L) indicated an in vitro activity comparable to that of vancomycin in methicillin-resistant Staphylococcus aureus (4 mg/L), S. epidermidis (2 mg/L) and methicillin-susceptible strains. Enterococcus faecalis strains were susceptible to linezolid (MIC90 2-4 mg/L), glycopeptides and beta-lactams. In E. faecium, only glycopeptides (MIC90 2 mg/L) and linezolid (MIC90 2 mg/L) were active. Linezolid was the only drug active against two strains of Enterococcus showing a VanA phenotype. Owing to its antibacterial profile, linezolid represents a promising drug for the treatment of Gram-positive infections. PMID- 12084109 TI - Abstracts of the British Association of Dermatologists 82nd Annual Meeting. 9-12 July 2002, Edinburgh, United Kingdom. PMID- 12084107 TI - Single- and multistep selection study of the antipneumococcal activity of BMS 284756 compared to ciprofloxacin, levofloxacin, trovafloxacin and moxifloxacin. AB - Single- and Multi-step selection studies were used to test the ability of BMS 284756, ciprofloxacin, levofloxacin, trovafloxacin and moxifloxacin to yield resistant clones from 12 quinolone-susceptible and -resistant Streptococcus pneumoniae strains. Although all quinolones selected, to a greater or lesser degree, for resistant clones with mutations usually in parC or gyrA, BMS-284756 tended to select for resistant clones at a lower rate than other quinolones studied. PMID- 12084108 TI - Detection of Kaposi's sarcoma herpesvirus DNA sequences in pericardial fluid in a patient with multicentric Castleman's disease. PMID- 12084122 TI - Global ramifications of European alcohol policies. PMID- 12084123 TI - Brief interventions over the long term: unfinished business. PMID- 12084124 TI - A review of the published literature into cannabis withdrawal symptoms in human users. AB - Recent experimental papers have been published suggesting the appearance of withdrawal symptoms upon the cessation of cannabis use in human users and proposing the introduction of a diagnostic category for such symptoms. Research also continues to be published into the physiological effects of cannabis on animals via self-administration paradigms and the use of cannabinoid antagonists. Animal research does not provide a clear picture of a consistent withdrawal effect. The literature on withdrawal symptoms appearing in human users following the cessation of cannabis is investigated in this paper to clarify this issue further and enhance the scientific and lay debate on the status of the drug. Methodological weaknesses in the literature are highlighted. These include variable levels of drug-dose administered in laboratory conditions, lack of controlled studies and the absence of definitions of the withdrawal syndrome sought. It is suggested that the studies conducted to date do not provide a strong evidence base for the drawing of any conclusions as to the existence of a cannabis withdrawal syndrome in human users, or as to the cause of symptoms reported by those abstaining from the drug. On the basis of current research cannabis cannot be said to provide as clear a withdrawal pattern as other drugs of abuse, such as opiates. However, cannabis also highlights the need for a further defining of withdrawal, in particular the position that rebound effects occupy in this phenomenon. It is concluded that more controlled research might uncover a diagnosable withdrawal syndrome in human users and that there may be a precedent for the introduction of a cannabis withdrawal syndrome before the exact root of it is known. PMID- 12084125 TI - Alcohol dependence: a public health perspective. AB - AIMS: To review epidemiological research on alcohol dependence and to propose a public health approach to address this problem. DESIGN AND SETTING: The research under review is basically constituted by general population surveys. PARTICIPANTS: Individuals in the community 18 years of age and older. MEASUREMENT: Twelve-month rates of alcohol dependence are estimated using DSM criteria. FINDINGS: The prevalence of alcohol dependence has been stable in the United States, at around 6% for men and 2% for women. The risk of alcohol dependence begins at low levels of drinking and increases linearly with alcohol consumption. Mean group level of consumption shows some relationship to prevalence of dependence. Alcohol-dependent individuals are responsible for about 50% of the social, legal and interpersonal alcohol-related problems in society. CONCLUSION: Alcohol dependence is present at relatively high levels in the community. Alcohol-dependent individuals are responsible for a fair proportion of alcohol-related problems in society. A public health approach to this problem, emphasizing prevention and group level interventions to lower prevalence levels, is warranted. PMID- 12084127 TI - Executive summary of the National Research Council's Report 'Informing America's policy on illegal drugs: what we don't know keeps hurting us'. PMID- 12084128 TI - The first agenda for drug enforcement research. PMID- 12084129 TI - Time to be 'confused' by facts? PMID- 12084130 TI - Drug policies: improving the quality of the relevant debate. PMID- 12084131 TI - Matching drug policy research to drug policy goals. PMID- 12084132 TI - A report that should become a well-thumbed desk companion. PMID- 12084133 TI - The best should not be the enemy of the good. PMID- 12084134 TI - What we don't want to know keeps hurting us. PMID- 12084136 TI - The long-term effectiveness of brief interventions for unsafe alcohol consumption: a 10-year follow-up. AB - AIMS: To examine the long-term impact of brief and early interventions for hazardous and harmful alcohol consumption. DESIGN: A 9-month and 10-year follow up of subjects recruited into a randomized controlled trial of a range of alcohol related brief interventions. SETTING: General practices, the outpatient or acute care services of a major city hospital, and a privately run health screening programmeme. PARTICIPANTS: The cohort of 554 (non-dependent) hazardous and harmful drinkers recruited into the Australian arm of the Phase II World Health Organization collaborative project on identification and treatment of persons with harmful alcohol consumption. INTERVENTION: The effectiveness of three forms of intervention, ranging from 5 to 60 minutes in duration, were compared with a no-treatment control condition. MEASUREMENTS: Included drinking behaviour and biological markers of alcohol use. In addition, at 10 years subjects were asked about symptoms of diagnosable alcohol use disorders and their experience of alcohol-related psychological, social and physical harm. Mortality was also assessed. FINDINGS: Results provide further evidence for the short-term effectiveness of alcohol-related brief interventions. In comparison to controls, subjects offered intervention: (1) report significantly lower consumption; and (2) less unsafe drinking at 9-month follow-up. The intensity of intervention was not related to the amount of change in drinking behaviour. Analysis at 10 years failed to find any differences in outcomes between intervention and control groups in median consumption, mean reduction in consumption from baseline to follow-up, mortality and ICD-10 diagnoses of alcohol dependence or harmful alcohol use. CONCLUSIONS: This study failed to find evidence that brief advice and counselling without regular follow-up and reinforcement can sustain significant long-term reductions in drinking behaviour at 10-year follow-up. PMID- 12084137 TI - Process and outcome changes with relapse prevention versus 12-Step aftercare programs for substance abusers. AB - AIMS: Presumptive support was sought for mechanisms of action whereby two conceptually distinct aftercare programs, relapse prevention (RP) and 12-Step facilitation (TSF), impact upon substance abusers. PATIENTS AND DESIGN: Adults who had just completed intensive treatment were assigned randomly to either RP (n=61) or TSF (n=70) aftercare programs. SETTING: Three residential treatment facilities. INTERVENTIONS: Trained counselors delivered to small groups a manualized aftercare program which focused either upon the utilization of cognitive-behavioral processes to orchestrate change through an individualized treatment plan (i.e. RP) or which sought to facilitate utilization of AA's 12 Steps (i.e. TSF). MEASUREMENTS: Process measures developed specifically to quantify either: (a) the changes in self-efficacy process in RP or (b) the utilization of AA's principles in TSF, as well as psychosocial and substance abuse indices were administered to all patients pre- and post-aftercare and at 6 month follow-up. FINDINGS: A significant relationship between changes in measures of self- efficacy for RP participants as well as a trend for a relationship between process-specific change for TSF participants partially satisfied the first condition for presumptive support. The fact that the intervention-specific mediators covaried with several outcome indices, and that removal of such mediators attenuated prediction of outcome met, respectively, the second and third conditions for presumptive support. CONCLUSION: Carefully orchestrated RP and TSF aftercare programs yield process changes that are related positively to improved outcome. PMID- 12084138 TI - A randomized trial of a brief alcohol intervention for needle exchangers (BRAINE). AB - AIMS: To test motivational interviewing (MI) as a brief intervention for reducing alcohol use among needle exchange clients. DESIGN: Randomized clinical trial. SETTING: Needle exchange program-Providence, Rhode Island, USA. PARTICIPANTS: Between 2/98 and 10/99, we recruited 187 AUDIT-positive (>8) active injection drug users. INTERVENTION: Those assigned to a brief motivational intervention (MI) condition received two 1-hour therapist sessions following assessment visits, 1 month apart, focusing on alcohol use and HIV risk-taking. MEASUREMENTS: Control and MI subjects received identical research assessments at baseline, 1 and 6 months following study enrollment. At 6 months, study outcomes included days of alcohol use measured using the time-line follow-back method. FINDINGS: Study retention was 96.8% at 6 months. Participants reported an average of 12.0 drinking days at baseline and 8.3 at 6 months. Significant reductions in drinking days were observed in both treatment conditions. We found significant treatment x baseline drinking day interaction effects. Tests for simple main effects were significant for subjects with above median (>9) baseline drinking day frequency, but not for those with below median baseline drinking frequency. Comparisons on dichotomous outcomes provided supporting evidence of treatment efficacy; those in MI were over two times more likely than controls to report reductions of 7 days or more (P < 0.05). CONCLUSIONS: This study provides the first direct evidence that brief MI can decrease alcohol use among active injection drug users with drinking problems. Heavier drinkers seem best suited for this intervention, but the optimal intensity of treatments and which components of brief intervention are most effective deserve further study. PMID- 12084139 TI - Whose drinking does the liberalization of alcohol policy increase? Change in alcohol consumption by the initial level in the Finnish panel survey in 1968 and 1969. AB - AIMS: To study how the annual volume of alcohol consumed changed among population groups in different categories of initial consumption in a situation where per capita consumption strongly increased due to a new alcohol law. DESIGN: A post hoc quasi-experiment with a two-wave panel study. The regression to the mean effect was accounted for by using reference data from two auxiliary panel studies. SETTING: Finland in 1968 and 1969, when per capita consumption increased by 46% in a year. Reference data come from Norway in 1975 and 1976, and the United States in 1993 and 1994. PARTICIPANTS: General population sample of 1720 (1292 men) aged 15-69 (response rate 91.4%). REFERENCE DATA: General population surveys in a Norwegian town (n=254) and in the United States (n=658). MEASUREMENTS: Annual volume of alcohol consumed. FINDINGS: In Finland, the greatest change in observed consumption levels occurred among those with moderate consumption levels, but the same phenomenon was observed in the reference data. A comparison between the Finnish and the reference data indicated that in Finland consumption increased at all levels, and the higher the initial consumption level, the greater the increase. CONCLUSIONS: A best estimate for the differential impact of future increases in alcohol consumption is that moderate and particularly heavy drinkers will be affected to a greater extent than will light drinkers or abstainers. PMID- 12084140 TI - Beyond the U-curve: the relationship between sport and alcohol, cigarette and cannabis use in adolescents. AB - AIMS: This study aimed at increasing understanding of the relationship between sporting activities and alcohol, cigarette and cannabis use among adolescents, by examining gender, age and the context of sporting practice. DESIGN, SETTING, SUBJECTS: The study was a national school survey (n=10807; ages 14-19 years) conducted in France in 1999. MEASURES: Respondents were asked confidentially by self-administered questionnaire (pen and paper) about their use of licit and illicit drugs and life-style (including sporting activities outside school: hours per week, registration in a club, type of sport). FINDINGS: The U-shaped curve between the intensity of physical activities and licit and illicit drug use appeared not to be systematic. It depended mainly on the product and the level of use. It only remained significant for boys and heavy smoking once gender and age effect were taken into account. CONCLUSION: The results stress the need to control for age and gender when the survey participants are teenagers. The relationship between drug use and sporting activity also depends on the type of sport. PMID- 12084141 TI - Educational and occupational attainment and drinking behavior: an expectancy model in young adulthood. AB - AIMS: The socio-economic status (SES) variables of education level and occupational functioning have been found to be correlated negatively with alcohol use. The present study examined prospectively the relationship between these functioning measures, alcohol expectancies and alcohol involvement. We propose that expectancies function as a mediator of the relationship between educational/occupational attainment and drinking behavior. We hypothesized that changes in young adult functioning are linked to changes in social context and/or the availability of non-alcohol reinforcers, which in turn affect the reinforcement expected from alcohol. PARTICIPANTS AND DESIGN: Participants were 172 young adults from an ongoing longitudinal study of long-term clinical course of adolescent substance use treatment. Data from 6- and 8-year follow-ups were used in the present analyses. The treated sample (n=100) was recruited from in patient substance abuse treatment programs for adolescents. A community sample (n=72) was matched on family history of substance abuse and SES at intake. FINDINGS: A cross-lag panel analysis indicated that education had a unique longitudinal relationship with expectancy for both the treated and community sample, over and above previous alcohol use and expectancy. Occupational variables did not have a longitudinal relationship with alcohol use and expectancy for either sample. Expectancies mediated the education/drinking relationship for the treated sample only. CONCLUSIONS: These results suggest one means through which changes in functioning may alter alcohol involvement over time: alteration of the reinforcement expected from alcohol. PMID- 12084142 TI - Determining a diagnostic cut-off on the Severity of Dependence Scale (SDS) for cocaine dependence. AB - AIM: The study aimed to assess the efficacy of the Severity of Dependence Scale (SDS) as a diagnostic measure of cocaine dependence and determine the cut-off score that best discriminates between the presence and absence of a DSM-IV diagnosis of cocaine dependence. DESIGN: Cross-sectional survey. SETTING: Sydney, Australia. PARTICIPANTS: One hundred and forty-two cocaine users. MEASUREMENTS: The diagnostic performance of the SDS was measured via ROC analysis against DSM IV diagnoses of cocaine dependence, as measured by the Composite International Diagnostic Interview (CIDI). FINDINGS: ROC analysis revealed the SDS to be a test of high diagnostic utility for the measurement of cocaine dependence. The cut-off point on the SDS at which there is optimal discrimination between the presence and absence of a DSM-IV diagnosis of cocaine dependence was found to be 3 (i.e. a score of 3 or more). CONCLUSIONS: The study statistically validated the utility of the SDS as a diagnostic measure of cocaine dependence and has determined an appropriate cut-off point. The SDS is recommended as a brief screening instrument for cocaine dependence that can be used in addition to more comprehensive measures, such as the CIDI. PMID- 12084143 TI - Gender differences in the psychological determinants of cigarette smoking. AB - AIMS: To compare the distributions of smoking-related variables and the size of associations between these variables in men and women. DESIGN AND PARTICIPANTS: Mail survey in 2934 daily smokers (1533 women and 1401 men) who volunteered for a smoking cessation trial. Follow-up after 7 months in 2456 people (84%). SETTING: Community setting (French-speaking part of Switzerland, 1998). FINDINGS: Women smoked less than men (18 versus 22 cigarettes per day, p < 0.001), had lower confidence in their ability to refrain from smoking, used more frequently the strategy defined as 'coping with the temptation to smoke' and reported more drawbacks of smoking (gender differences ranged between 0.1 and 0.3 standard deviation units on these scales). There was no gender difference in the distribution of smokers by stage of change. At follow-up, smoking cessation rates were similar in men and women (6% versus 5%, p=0.3). Intention to quit, quit attempts in the previous year and a more frequent use of self-change strategies predicted smoking cessation and were associated with tobacco dependence in both sexes. A more frequent use by women of coping strategies suggests that some women are 'self-restrained' smokers who control their smoking permanently. This could explain lower smoking rates in women. The size of associations between smoking related variables was similar in men and women. CONCLUSIONS: Even though there were gender differences in the distributions of some smoking-related variables, associations between these variables were similar in men and women. This suggests that smoking behaviour is regulated by similar psychological mechanisms in men and women. PMID- 12084144 TI - Cigarette smoking is associated with increased severity of gambling problems in treatment-seeking gamblers. AB - AIMS: Cigarette smoking and gambling often co-occur, but very little is known about smoking or its correlates in disordered gamblers. This study compared gambling and psychosocial problems in treatment-seeking gamblers who smoke versus those who do not. METHODS: At intake to gambling treatment programs, gamblers completed the addiction severity index (ASI) and gambling questionnaires. Subjects were categorized into non-daily smokers (n=107) and daily smokers (n=210). Differences in demographics, gambling variables and ASI composite scores were compared between the groups. RESULTS: The daily smokers were more likely to have a history of treatment for a substance use disorder than the non-daily smokers. After controlling for substance abuse treatment histories, gender and age, the daily smokers demonstrated more severe gambling, family/social and psychiatric problems. Compared to non-daily smokers, the daily smokers gambled on more days and spent more money gambling; they also 'craved' gambling more and had lower perceived control over their gambling. The daily smokers were more likely to be taking psychiatric medications, and they experienced psychiatric symptoms, especially anxiety symptoms, on a greater number of days than non-daily smokers. IMPLICATIONS: Results from this study suggest that about two-thirds of treatment seeking gamblers are current daily cigarette smokers, and smoking status is associated with more severe gambling and psychiatric symptoms. These results warrant further investigation of smoking in gamblers and whether smoking adversely affects the course of treatment or outcomes among gamblers. PMID- 12084145 TI - Getting the cannabis withdrawal question into proper perspective: comments on Smith. PMID- 12084146 TI - Cannabis produces dependence: a comment on Smith. PMID- 12084147 TI - Defining the indefinable: comments on Smith. PMID- 12084148 TI - Addiction's decision to withdraw a published paper from citation on the grounds of undisclosed conflict of interest. PMID- 12084157 TI - Animal models of bovine spongiform encephalopathy and vCJD infectivity in blood: two swallows do not a summer make. PMID- 12084158 TI - Brain and buffy coat transmission of bovine spongiform encephalopathy to the primate Microcebus murinus. AB - BACKGROUND: More than 100 cases of variant CJD resulting from infections with bovine spongiform encephalopathy (BSE) have accumulated in the United Kingdom since 1995. Concern about the possibility of secondary transmissions via blood and blood components donated by infected individuals has prompted a variety of international donor deferral policies that will continue until laboratory and epidemiologic evidence provides a consensus about potential risk. STUDY DESIGN AND METHODS: BSE was passaged through macaque monkeys and then adapted to the prosimian microcebe (Microcebus murinus). Brain homogenate and buffy coat from an affected microcebe were separately inoculated intracerebrally into three healthy microcebes (two animals received brain and one received buffy coat). RESULTS: All three inoculated microcebes became ill after incubation periods of 16 to 18 months. Clinical, histopathologic, and immunocytologic features were similar in each of the recipients. CONCLUSION: Buffy coat from a symptomatic microcebe infected 17 months earlier with BSE contained the infectious agent. This observation represents the first documented transmission of BSE from the blood of an experimentally infected primate, which in view of rodent buffy coat infectivity precedents and the known host range of BSE is neither unexpected nor cause for alarm. PMID- 12084159 TI - Comparative analysis of normal prion protein expression on human, rodent, and ruminant blood cells by using a panel of prion antibodies. AB - BACKGROUND: It is not known whether variant CJD can be transmitted within the human population by blood transfusion. The expression of normal cellular prion protein (PrPC) by different blood cell types may permit selective uptake and dissemination of infectivity. STUDY DESIGN AND METHODS: The normal distribution of PrPC on the major blood cell types of species known to be susceptible to natural or experimental transmissible spongiform encephalopathies was studied. Blood from healthy humans, mice, hamsters, cattle, and sheep was examined by flow cytometry by using a large panel of antibodies with different prion protein (PrP) epitope specificities to maximize the detection of PrP variants across species and cell type. RESULTS: PrP was detected on all major human blood cells types except eosinophils, but was not detected as ubiquitously or uniformly on major blood cell types of different animal species. CONCLUSION: Different animal species have unique patterns of expression of PrPC on blood cell types, with none equivalent to the human pattern. This needs to be considered when extrapolating from animal models of blood-borne transmissible spongiform encephalopathy infectivity, particularly in regard to the risk assessment of potential variant CJD spread within the human population. The relationship between PrP distribution and infectivity distribution in blood needs further investigation. PMID- 12084161 TI - Mathematic modeling of the risk of HBV, HCV, and HIV transmission by window-phase donations not detected by NAT. AB - BACKGROUND: Blood transfusion centers around the world have introduced minipool NAT to reduce the risk of HBV, HCV, and HIV transmission by blood donations drawn in the infectious window phase. What would be the reduction in the residual risk when minipool NAT would be replaced by single-donation NAT? STUDY DESIGN AND METHODS: A mathematic model was developed to estimate the probability of virus transmission by blood transfusion when NAT screening methods are used for virologic safety testing. The major assumptions used are threefold: 1) The viral nucleic acid concentrations in the early window phase of infection double in 2.8 (HBV), 0.74 (HCV), and 0.90 (HIV) days. 2) The detectability of low copy numbers of viral DNA or RNA by the screening assay can be described with a probit model. 3) The probability of infection depends linearly on the logarithm of the administered dose, with 50-percent infectivity rates at 10 (HBV and HCV) or 1000 (HIV) viral nucleic acid copies per transfusion unit (estimates based on NAT studies with samples of known infectivity in chimpanzees). RESULTS: A reasonably simple equation was obtained that allows studying the effect of the sensitivity of the NAT assay and of the pool size used for screening on the residual risk of transfusion-transmitted infection. The computations are illustrated by using observed sensitivity estimates of various NAT methods. By using epidemiologic data among European donors over 1997 as baseline, the calculations predict that the incidence of virus transmission per 10-million RBC transfusions reduces with the following numbers when lowering the test pool size from 96 to 1 (single donation testing): HBV from 11 to 13 to 3.3 to 5.1, HCV from 1.7 to 2.0 to 0.5 to 0.8, and HIV from 0.47 to 0.62 to 0.010 to 0.045 (ranges for the different NAT screening methods). CONCLUSION: A proper mathematic model for the calculation of residual infection risk by blood transfusion helps understand the impact of introducing new NAT methods for blood safety testing. PMID- 12084160 TI - Sensitivity of HCV RNA and HIV RNA blood screening assays. AB - BACKGROUND: The FDA requirement for sensitivity of viral NAT methods used in blood screening is a 95-percent detection limit of 100 copies per mL, whereas the NAT screening system should have a sensitivity of at least 5000 copies per mL per individual donation. According to the Common Technical Specifications of the European Directive 98/79/EC for in vitro diagnostics, viral standard dilutions (calibrated against the WHO standard) should be tested at least 24 times for a statistically valid assessment of the 95-percent detection limit. STUDY DESIGN AND METHODS: Viral standard dilution panels (PeliCheck, VQC-CLB) were prepared for HCV RNA genotypes 1 and 3 and for HIV RNA genotypes B and E. In a multicenter study, 23 laboratories tested the panels all together in 8 to 91 test runs per NAT method. RESULTS: The following 95-percent detection limits (and 95% CIs) were found on the HCV RNA genotype 1 reference panels (shown as geq/mL): Gen-Probe TMA, 85 (64-118); AmpliScreen, 126 (83-225); AmpliScreen with NucliSens Extractor, 21 (13-44); Amplicor with NucliSens Extractor, 69 (50-102), and Amplicor with Qiagen extraction technology, 144 (74-102). On HIV RNA genotype B dilution panels, the following 95-percent detection limits were found (shown as geq/mL): Gen-Probe TMA, 31 (20-52); AmpliScreen, 126 (67-311); AmpliScreen with NucliSens Extractor, 37 (23-69), and NucliSens QL assay, 123 (51-566). HIV RNA genotype E panels were detected with equal sensitivity as HIV RNA genotype B panels. In the Gen-Probe TMA assay, the 50-percent detection limits on HIV RNA type B and type E were 3.6 (2.6-5.0) and 3.9 (2.4-5.8) geq per mL, respectively. The HCV RNA genotype 1 and 3 standards were detected with equal sensitivity. CONCLUSION: The differences in sensitivity between NAT assays can be explained by the input of isolated viral nucleic acid in the amplification reactions. The FDA requirements for sensitivity of NAT blood screening assays can be met by the Gen probe TMA, as well as by the AmpliScreen assays, particularly when combined with the NucliSens Extractor. PMID- 12084162 TI - Trypanosoma cruzi in Los Angeles and Miami blood donors: impact of evolving donor demographics on seroprevalence and implications for transfusion transmission. AB - BACKGROUND: Trypanosoma cruzi, the agent of Chagas' disease, continues to be a concern for blood safety, as demonstrated by recent transfusion-transmitted cases in the United States and Canada. The chronic nature of Chagas', coupled with increasing numbers of immigrants from T. cruzi-endemic countries, suggests that Chagas' is a long-term public health problem. Herein, we report on a multiyear epidemiologic study of T. cruzi in Los Angeles and Miami blood donors. STUDY DESIGN AND METHODS: From May 1994 to September 1998, blood donors in Los Angeles and Miami were queried regarding birth or time spent in an endemic country. Donations of "yes" respondents were tested by EIA, confirmed by radioimmunoprecipitation assay, and if confirmed as seropositive, enrolled in look-back investigations. RESULTS: A total of 1,104,030 Los Angeles and 181,139 Miami donors were queried regarding risk; 7.3 and 14.3 percent, respectively, responded yes. Seropositive rates were 1 in 7,500 Los Angeles and 1 in 9,000 Miami donors. In Los Angeles, seroprevalence rates increased significantly from 1996 to 1998 and were significantly higher for directed donors than nondirected donors. Look back identified 18 recipients, all of whom were seronegative for T. cruzi. CONCLUSION: Significant numbers of T. cruzi-seropositive donors contribute to the U.S. blood supply. The incidence of seropositivity is enhanced by minority recruitment efforts necessitated by donor demographic shifts. Similarly, high rates among directed donations in Los Angeles are attributable to a disproportionate number of at-risk directed donors. Current look-back data likely underestimate the frequency of transfusion- transmitted T. cruzi. These results indicate that continued evaluation of transfusion as a mode of T. cruzi transmission in the United States is needed. PMID- 12084164 TI - Transfusion support with RBCs from an Mk homozygote in a case of autoimmune hemolytic anemia following diphtheria-pertussis-tetanus vaccination. AB - BACKGROUND: Autoimmune hemolytic anemia (AIHA) in children, although unusual, is often associated with recent infection. Several reports have identified the diphtheria-pertussis-tetanus (DPT) vaccination as a possible trigger for AIHA. STUDY DESIGN AND METHODS: Life-threatening AIHA was diagnosed in a 6-week-old infant 5 days after receiving a DPT vaccination. The patient required daily transfusion and/or exchange transfusion for 3 weeks. RBCs from an Mk homozygote were found compatible with the patient's autoantibody. Transfusion of RBCs from an Mk homozygote and later RBCs from an individual (K.T.) with a variant glycophorin, Mi.VII, were required to sustain the patient's Hb level until autoantibody production ceased, as evidenced by a fall in antibody titer and the patient's Hct returning to normal. RESULTS: The DAT was positive (3+) with only anti-C3 on presentation. An IgM cold reactive autoantibody with probable anti-Pr specificity and high thermal amplitude (37 degrees C) was identified in the serum. The DAT was no longer positive after transfusion with compatible blood. CONCLUSION: This case represents life-threatening AIHA in an infant, temporally related to a DPT injection and responsive to a combination of immunosuppression and transfusion of rare compatible blood. PMID- 12084163 TI - A randomized controlled trial comparing the frequency of acute reactions to plasma-removed platelets and prestorage WBC-reduced platelets. AB - BACKGROUND: Removal of the plasma supernatant from platelets before transfusion is effective in preventing acute reactions to platelets caused by cytokines. Prestorage WBC reduction of platelets may be even more effective at preventing reactions as the WBCs are removed and WBC-derived cytokines do not accumulate in this component. This study evaluates the effectiveness of plasma removal and two methods of prestorage WBC reduction for preventing acute reactions to platelets. STUDY DESIGN AND METHODS: Platelets given to adults with hematologic malignancies were randomly allocated to one of three types: plasma supernatant removed and a platelet storage solution added, whole blood-derived platelets that are prestorage WBC reduced by filtration before storage, and prestorage WBC-reduced apheresis platelets. Patients were monitored before, during, and after transfusion, and the severity of reactions was graded on a Likert scale. RESULTS: A total of 129 patients from four centers were given 1190 platelet transfusions. The overall frequency of reactions was 13.6 percent (162 of 1190), 21.3 percent (36 of 169) for the plasma-removed platelets, 11.4 percent (59 of 517) for random donor WBC-reduced platelets, and 13.3 percent (67 of 504) for apheresis WBC reduced platelets (p=0.384). The overall frequency of severe reactions was 4.1 percent with plasma-removed platelets, 1.7 percent for whole blood-derived, prestorage WBC-reduced platelets, and 1.4 percent for prestorage WBC-reduced apheresis platelets. CONCLUSION: The frequency of reactions to plasma-removed platelets and prestorage WBC-reduced platelets was not significantly different; however, the power of the study for this comparison was low. There was no difference in the frequency of reactions to the two types of prestorage WBC reduced platelets. The frequency of severe reactions to prestorage WBC-reduced platelets is low, occurring in only 1 to 2 percent of transfusions. PMID- 12084165 TI - Predicting response to plasma exchange in patients with thrombotic thrombocytopenic purpura with measurement of vWF-cleaving protease activity. AB - BACKGROUND: Severe deficiency of vWF-cleaving protease (vWF-CPase) activity was recently found in patients with thrombotic thrombocytopenic purpura (TTP). Although the survival of patients with TTP has been dramatically improved with plasma exchange (PE), there are still many patients who are refractory to PE and immunosuppressive therapy. STUDY DESIGN AND METHODS: The activities of vWF-CPase and its inhibitor were measured in 27 patients with nonfamilial TTP and hemolytic uremic syndrome (HUS) to examine the relationship between the clinical variables and vWF-CPase activity. RESULTS: Eight of nine patients with HUS had more than 40 percent of vWF-CPase activity, whereas one had 28 percent of the normal level at the acute phase. Ten of 12 TTP patients with a good outcome had a severe deficiency of vWF-CPase activity and its inhibitor, whereas four of six patients with a poor outcome had a moderate deficiency of vWF-CPase activity along with a lack of the inhibitor. PE produced normalization of the vWF-CPase activity and neutralization of the inhibitor in TTP patients with a good outcome; however, some TTP patients with vWF-CPase inhibitor had relapsed and required an immunosuppressive therapy. The response to the combination therapy with PE and immunosuppressive treatment was poor in TTP patients without a severe deficiency of vWF-CPase activity. CONCLUSION: Assays of vWF-CPase activity and its inhibitor may be useful for predicting the response to therapy and the outcome of patients with TTP. In some patients, nonfamilial TTP with a poor prognosis may not be caused by a constitutional or acquired deficiency of vWF-CPase with its inhibitor. Although PE and immunosuppressive therapy are effective in patients with nonfamilial TTP and a vWF-CPase inhibitor, other therapeutic modalities may be needed for nonfamilial TTP with unknown etiology. PMID- 12084166 TI - Ex vivo expansion of long-term culture initiating marrow cells by IL-10, SCF, and IL-3. AB - BACKGROUND: Ex vivo expansion of progentior cells may shorten hematopoietic regeneration after myeloablative chemoradiotherapy, increase target cells for gene therapy, and improve purging of progenitor cell components. STUDY DESIGN AND METHODS: Marrow cells were incubated for 1 week in suspension culture with and without IL-10, IL-3, and SCF. As long-term culture initiating cells (LTC-ICs) represent early hematopoietic progenitors in vitro, these cells were quantified at initiation and after a 1-week culture period in a limiting dilution assays. Additionally, immunophenotyping of cells before and after culture was performed. RESULTS: In six experiments, marrow cells cultured for 1 week with IL-10, IL-3, and SCF showed a significant increase (almost doubling) in LTC-ICs as compared with marrow cells before expansion. Additionally, an increased proliferative capacity of LTC-ICs was achieved with a sevenfold increase of committed colony forming cells and a 10-fold proliferation of high proliferative potential colony forming cells. Immunophenotyping revealed a sevenfold increase of CD34+ CD45 RA- cells in IL-10-, IL-3-, SCF-stimulated suspension cultures. In unstimulated cultures, no LTC-ICs were maintained after 1 week. CONCLUSION: Expansion of LTC ICs by IL-10, IL-3, and SCF has not been shown so far. This in vitro model allows expansion of LTC-IC if compared with the input of progenitor cells without extensive progenitor cell manipulation. This should be an attractive model for in vitro purging, gene transfer, or expansion of progenitor cells to allow rapid engraftment after myeloablative chemotherapy. PMID- 12084167 TI - Matrix metalloproteinase-9 (gelatinase B) is elevated during mobilization of peripheral blood progenitor cells by G-CSF. AB - BACKGROUND: Matrix metalloproteinase-9 (MMP-9 or gelatinase B) has recently been implicated in the IL-8-induced mobilization of HPCs in rhesus monkeys and mice. It is not known whether administration of G-CSF causes expression of MMP-9 during HPC mobilization. STUDY DESIGN AND METHODS: Blood samples from 15 allogeneic progenitor cell donors were collected before and during G-CSF-induced HPC mobilization. The expression of the gelatinases MMP-2 and MMP-9 in the plasma of the donors was analyzed by ELISA and zymographic analysis. Gelatinolytic activity was measured with a fluorometric assay that was specific for gelatinases. Expression of IL-6, IL-8, and soluble vascular cell adhesion molecule (VCAM) was measured by ELISA. RESULTS: Highly elevated latent gelatinolytic activity was found on Days 4 and 5 of G-CSF treatment in comparison to pretreatment activity. ELISA and zymographic analyses revealed pro-MMP-9 as the major source of the latent gelatinolytic plasma activity during mobilization. Pro-MMP-2 was not elevated compared with pretreatment levels. As IL-8 has been implicated in the expression of MMP-9, IL-8 concentrations were measured in plasma samples from donors and patients immediately before the start of HPC apheresis, but no significantly elevated IL-8 concentrations were noted. In contrast, pro-MMP-9 and latent gelatinolytic activity was highly correlated with IL-6, which was strongly elevated during mobilization therapy. Finally, soluble VCAM was equally significantly elevated on the days of apheresis. CONCLUSIONS: G-CSF mobilization treatment induces MMP-9, IL-6, and soluble VCAM. Expression of MMP-9 might be involved in the mobilization of human HPCs and might be a final common pathway of different mobilization therapies. Our data do not support a role of IL-8 in G-CSF induced mobilization. In contrast, IL-6 might be involved in the G-CSF-induced expression of MMP-9. PMID- 12084168 TI - Kinetics of G-CSF-induced granulocyte mobilization in healthy subjects: effects of route of administration and addition of dexamethasone. AB - BACKGROUND: Granulocyte donors are frequently given G-CSF with or without dexamethasone approximately 18 hours before apheresis to increase cell yields. The purpose of this study was to assess the kinetics of G-CSF plus dexamethasone neutrophil mobilization to determine whether the neutrophils can be mobilized and collected the same day. STUDY DESIGN AND METHODS: Sixteen subjects were given four separate mobilization regimens: IV G-CSF (5 microg/kg), subcutaneous G-CSF (5 microg/kg), IV G-CSF (5 microg/kg) plus oral dexamethasone (8 mg), and subcutaneous G-CSF (5 microg/kg) plus oral dexamethasone (8 mg). Blood cell counts were measured before and after G-CSF administration. RESULTS: Following all four mobilization regimens, neutrophil counts fell 0.5 hour after the mobilizing agents were given, rose above baseline levels at Hour 2, and increased further with each time interval to Hour 8. In the absence of dexamethasone at Hours 2 through 8, there was no difference in neutrophil counts by subcutaneous or IV G-CSF administration routes. The addition of dexamethasone enhanced mobilization of neutrophils from Hours 3 through 24. Through Hour 8, there was no difference in the degree of mobilization among the subcutaneous G-CSF plus dexamethasone and the IV G-CSF plus dexamethasone regimens. However, at Hour 24, neutrophil counts were sustained at higher levels with subcutaneous G-CSF plus dexamethasone than with IV G-CSF plus dexamethasone. CONCLUSIONS: Granulocyte mobilization response to subcutaneous G-CSF plus dexamethasone is sustained at peak levels for 8 to 24 hours after coadministration of the two drugs. There was no advantage to giving G-CSF intravenously. PMID- 12084169 TI - Characterization and functional analysis of granulocyte concentrates collected from donors after repeated G-CSF stimulation. AB - BACKGROUND: Neutropenic patients often develop bacterial or fungal infections not responding to broad-spectrum antibacterial or antifungal agents. Clinical efforts were made with transfusion of granulocyte concentrates; however, functions of granulocytes after multiple G-CSF stimulations and after apheresis are not yet investigated and described sufficiently. STUDY DESIGN AND METHODS: The aim of this study was to characterize functional and immunologic variables of granulocytes in blood samples drawn from donors before and after each stimulation episode with G-CSF, in the resulting granulocyte concentrates and in the patients 8 hours after transfusion. RESULTS: Chemotaxis was not influenced, neither by G CSF application nor by apheresis. Multiple G-CSF stimulations enhanced oxidative burst and phagocytosis of Escherichia coli in donor granulocytes. These values returned to basal levels in granulocyte concentrates. Expression of granulocytic surface antigens was downregulated after application of G-CSF but returned to normal and in part enhanced values in concentrates. A clinically relevant increase of proinflammatory cytokines could not be detected. Leukotriene B4 production was reduced after the fourth G-CSF stimulation in the donor blood and enhanced in the granulocyte concentrate after apheresis. Results in recipients indicate that changes of granulocyte function noted in concentrates were only transient. CONCLUSION: Stimulation of healthy donors with repeated G-CSF injections and subsequent granulocyte apheresis does not dramatically change decisive functions of granulocytes. PMID- 12084170 TI - A newly developed gel centrifugation test for quantification of RBC-bound IgG antibodies and their subclasses IgG1 and IgG3: comparison with flow cytometry. AB - BACKGROUND: Novel gel centrifugation test (GCT) cards were evaluated with respect to their ability to estimate the quantity of IgG on RBCs and the determination of the IgG subclasses IgG1 and IgG3. STUDY DESIGN AND METHODS: In 65 patients with a positive DAT, the amount of IgG-gamma-, IgG1, and IgG3 on RBCs was examined by use of GCT cards and flow cytometry (FC) in parallel. The results were correlated with the presence or absence of hemolysis. In addition, D+ RBCs were studied after sensitization with anti-D sera from 22 alloimmunized pregnant women. RESULTS: The amount of IgG on the RBCs as determined by GCT dilution cards correlated with FC (r=0.70, p < 0.0001). IgG subclass results as determined by GCT IgG subclass cards were confirmed by FC in 14 cases with an anti-IgG-gamma chain titer > or =300, whereas IgG subclass cards were not suitable in cases with anti-IgG-gamma-chain titers less than 300. In 44 patients with 2+ or 3+ DAT in the GCT and anti-IgG-gamma-chain titer < or =30, no hemolysis was observed, whereas hemolysis occurred in 13 of 14 patients with an anti-IgG-gamma-chain titer > or =300. GCT data obtained by IATs with anti-D sera were concordant with FC results. CONCLUSION: There is a correlation between the amount of RBC-bound IgG and immune hemolysis. The GCT cards that detect the anti-IgG-gamma-chain may be useful to predict hemolysis in patients with a 2+ or 3+ DAT in the GCT. The diagnostic value of GCT cards for IgG subclass testing should be investigated further. PMID- 12084171 TI - UV-visible spectrophotometric approach to blood typing II: phenotyping of subtype A2 and weak D and whole blood analysis. AB - BACKGROUND: A recently introduced quantitative blood typing approach uses antibody-induced changes in the UV-visible spectra of blood. Changes in the blood spectra's slope, caused by RBC agglutination, are translated into a numerical agglutination index (AI). Comparing the AI value against an established threshold yields a "yes and/or no" output from which to determine the phenotype. The efficacy and flexibility of this approach with whole blood use and the ability to analyze weak D, A2, and A2B were examined. STUDY DESIGN AND METHODS: Two hundred randomly selected blood bank donor samples were coded and forward typed directly from whole blood by using the spectrophotometric analysis. Reverse grouping on plasma from each sample was carried out with a new modified procedure by using higher ratios of plasma to RBCs. Results were compared to typing by an FDA cleared automated typing system. Twenty-seven weak D samples, 15 A2 and 12 A2B, were similarly analyzed from whole blood. PEG improved detection of weak D, A2 and A2B subtypes. RESULTS: All two hundred coded samples were accurately typed, yielding identical results to the blood bank analysis in both forward and reverse grouping. All the weak D samples and A2 and A2B samples were clearly identified, having AIs above the type threshold indicator value. CONCLUSION: Spectrophotometric blood typing successfully phenotyped ABO and D in 200 whole blood samples. Reverse grouping of plasma was equally successful. The same method can identify weak D and A2 and A2B subtypes. PMID- 12084172 TI - Molecular background of D(C)(e) haplotypes within the white population. AB - BACKGROUND: D(C)(e) and D(C)e haplotypes may be encountered in the white population. Few data are available on the molecular backgrounds responsible for depressed expression of C and e. STUDY DESIGN AND METHODS: Individuals of white origin carrying a D(C)(e) genotype resulting in depressed expression of C or both C and e were subdivided into two categories based on the RBC reactivity with the human sera Mol and Hor, which contain antibodies against low-frequency antigens of the Rh (RH) system and other non-Rh low-frequency antigens. Neither Hor+, Mol+ nor Hor+, Mol- RBCs expressed the V (RH10), VS (RH20), and/or Rh32 (RH32) low frequency antigens. These results suggested that Hor+, Mol+ variants expressed Rh33 (RH33 or Har) and FPTT (RH50), whereas Hor+, Mol- variants might express an undefined low-frequency antigen. Further serologic and molecular analyses were performed. RESULTS: Molecular analysis of Hor+, Mol+ variants revealed a hybrid gene structure RHCe-D(5)-Ce, in which exon 5 of RHCE (RHCe allele) was replaced by exon 5 of RHD (the so-called RHCeVA allele). The presence of exon 5RHD resulted in several amino acid alterations predicted in the external loop 4 of the CeVA polypeptide. Molecular analysis of Hor+, Mol- variants revealed the presence of a new RHCe allele characterized by a single point mutation C340T within exon 3 (the so-called RHCeMA allele), resulting in a R114W substitution predicted on the external loop 2 of the CeMA polypeptide. A serologic study showed a different pattern of reactivity with C and e MoAbs. CONCLUSION: Two types of mutations resulted in amino acid substitutions predicted in external loops 4 and 2, respectively, which altered both the C and e reactivity, and indicated conformation changes or defective interaction between nonadjacent loops of the Ce polypeptide. Serologic analysis showed that together with Hor and Mol sera testing, the use of different C and e MoAbs could help to identify these variants within the white population. PMID- 12084173 TI - RHC and RHc genotyping in different ethnic groups. AB - BACKGROUND: RH genotyping assays are mainly based on research in whites. These assays may not be reliable in a multiracial society because of the genetic variation in RH among ethnic groups. STUDY DESIGN AND METHODS: Five groups from different ethnic backgrounds were serologically typed for C and c and were genotyped on nucleotide C48 and intron 2 for RHC and RHc on nucleotides C178 and C307. RESULTS: RHc genotyping with both methods proved to be reliable. RHC genotyping on C48 is not reliable because of a 48G>C mutation in the RHce allele (false-positive prediction of C). This mutation was found in every ethnic group and does not affect c or e expression. RHC genotyping on intron 2 is unreliable because of r's (Cdes) alleles (a false-negative prediction of C). This allele was found in whites and blacks from Curacao and South Africa. Reactions of r's cells with anti-C are weaker, but no negative reactions with various MoAbs were found. A new method (RHC/c/hex3-intron 4/exon 7 multiplex PCRs) was developed based on intron 2 and r's hybrid exon 3 characteristics (RHC) and C307 (RHc). CONCLUSIONS: Reliable RHC and RHc genotyping is possible in different ethnic groups with the RHC/c/hex3-intron 4/exon 7 multiplex PCR approach. PMID- 12084174 TI - Novel 3'Rhesus box sequences confound RHD zygosity assignment. AB - BACKGROUND: Paternal RHD zygosity is required for genetic counseling and management of HDN caused by anti-D. The most common D- haplotype is due to the deletion of RHD, which results in the formation of a hybrid Rhesus box, theoretically through the recombination of 5' and 3'Rhesus boxes. STUDY DESIGN AND METHODS: The validity of Rhesus box PstI analysis was assessed to determine RHD zygosity by correlating D phenotype, most probable genotype, and Rhesus box PCR-RFLP. RHD exons were examined, and a 501-bp Rhesus box fragment was sequenced that flanked the polymorphic PstI site. RESULTS: Rhesus box analysis and the most probable genotype differed for 60 of 200 of the samples (30%). The incorrect zygosity assignment by the most probable genotype method is the likely reason for the difference. However, 8 of 328 samples showed Rhesus box copy numbers that were inconsistent with the D phenotype. Two D- samples with one hybrid Rhesus box had a nonfunctional RHD. Six D+ samples appeared to have two copies of the hybrid Rhesus box due to novel 3'Rhesus boxes that contained nucleotide polymorphisms previously assigned to the 5' and hybrid Rhesus boxes. All eight samples were from people of black descent, as determined by the GATA-1 silencing mutation at the FY locus. CONCLUSION: Rhesus box PCR-RFLP analysis for RHD zygosity assignment is confounded by the presence of nonfunctional RHD+ (2.3% of D-) and novel, low frequency (0.9% of all alleles) 3'Rhesus box sequences. PMID- 12084176 TI - Sickle cell patients deserve controlled recommendations. PMID- 12084175 TI - Human neutrophil antigen-2a expression on neutrophils from healthy adults in western Japan. AB - BACKGROUND: Human neutrophil antigen-2a (HNA-2a)-specific MoAb, which is necessary to prepare panel neutrophils and to determine the phenotype of patients' neutrophils, has been produced in many laboratories in Japan. The frequency of the HNA-2a-positive population in western Japan is unknown. STUDY DESIGN AND METHODS: A cell line secreting an HNA-2a-specific MoAb, TAG4, was established. TAG4 was characterized by flow cytometry, and the reactivities of TAG4 and another HNA-2a-specific MoAb, 7D8, were compared in an inhibition assay and two-color flow cytometry. TAG4 was tested against granulocytes from 408 healthy adult volunteers in western Japan. RESULTS: TAG4 reacted with neutrophils, but not with the other blood cells. The reactivity of TAG4 with granulocytes treated with glycosyl-phosphatidylinositol-specific phospholipase C was markedly reduced. The reactivity of TAG4 and 7D8 was similar, but 7D8 did not prevent TAG4 from binding. A total of 406 of 408 donors were TAG4 positive. The mean percentage of HNA-2a expression of females was 70.2 percent, and that of males was 66.8 percent. CONCLUSION: TAG4 and 7D8 seem to bind to different epitopes of HNA-2a. The frequency of the HNA-2a-positive population is 99.5 percent of healthy adults in western Japan. The mean percentage of HNA-2a expression of females was greater than in males. PMID- 12084178 TI - Efficacy of HBV NAT of pooled donor samples. PMID- 12084181 TI - Chair's Message--Dietitians of Canada's new values statement. PMID- 12084180 TI - A comparison of hospital readmission rates between two general physicians with different outpatient review practices. AB - BACKGROUND: There has been a relentless increase in emergency medical admissions in the UK over recent years. Many of these patients suffer with chronic conditions requiring continuing medical attention. We wished to determine whether conventional outpatient clinic follow up after discharge has any impact on the rate of readmission to hospital. METHODS: Two consultant general physicians with the same patient case-mix but markedly different outpatient follow-up practice were chosen. Of 1203 patients discharged, one consultant saw twice as many patients in the follow-up clinic than the other (Dr A 9.8% v Dr B 19.6%). The readmission rate in the twelve months following discharge was compared in a retrospective analysis of hospital activity data. Due to the specialisation of the admitting system, patients mainly had cardiovascular or cerebrovascular disease or had taken an overdose. Few had respiratory or infectious diseases. Outpatient follow-up was focussed on patients with cardiac disease. RESULTS: Risk of readmission increased significantly with age and length of stay of the original episode and was less for digestive system and musculo-skeletal disorders. 28.7% of patients discharged by Dr A and 31.5 % of those discharged by Dr B were readmitted at least once. Relative readmission risk was not significantly different between the consultants and there was no difference in the length of stay of readmissions. CONCLUSIONS: Increasing the proportion of patients with this age- and case-mix who are followed up in a hospital general medical outpatient clinic is unlikely to reduce the demand for acute hospital beds. PMID- 12084182 TI - Re: "Benefits and barriers associated with participation in food programs in three low-income Ontario communities". PMID- 12084183 TI - Food information programs: a review of the literature. AB - This paper summarizes existing evidence on the impact of food information programs. Published and unpublished literature produced within the past decade was searched and reviewed. Relevant data were tabulated and key findings summarized. Food information programs are becoming increasingly popular as tools to help consumers select a healthy diet. The key feature of a food information program is a package logo on foods meeting nutrition criteria set by the program s administering body. The logo acts as a health message. Several countries, including Canada, have adopted food information programs. Critics believe that these programs oversimplify the concept of healthy eating, that consumers misinterpret the logo s meaning, that licensing fees prohibit small companies from participating, and that the programs are limited to purchase behaviour and do not necessarily have an impact on dietary intake. Consumers report support for the programs and are able to interpret a logo s meaning accurately. In addition, evidence shows the programs have had a positive impact on the nutrient composition of foods. Research is still needed, however, to establish the impact of such programs on food purchases and dietary intake, and the overall and long term effectiveness of the programs as a nutrition intervention. PMID- 12084184 TI - Dietary acculturation and health-related issues of Indian immigrant families in Newfoundland. AB - Traditional Indian diets are high in grains, vegetables and fruits, and in this respect are substantially in accord with Canada s Food Guide to Healthy Eating. Adopting North American eating patterns carries potential health risks. In this study we explored migration-associated changes in eating patterns, lifestyle practices, and health-related issues of Indian immigrants and their families in Newfoundland. A cross-sectional survey employing a self-administered mailed questionnaire was conducted. A random sample of 132 subjects aged ten to 65+ took part. Participants were well established in Newfoundland and almost all were fluent in English. They were somewhat acculturated to foods commonly eaten in Canada. The majority reported that they were very or somewhat likely to engage in healthy lifestyle practices, and most (73%) wanted more relevant nutrition information. Consumption of grains, vegetables, and fruits was not in accordance with Canada s Food Guide to Healthy Eating. Sixty-five percent of participants reported that traditional Indian foods were not readily available, and 72% indicated a change in food-preparation methods. Studies of diverse ethnic groups are needed to guide effective nutrition education programs. University curricula and dietetic training programs should include cross-cultural courses to increase awareness of immigrants unique needs. PMID- 12084185 TI - Parkinson's disease: implications for nutritional care. AB - Parkinson's Disease (PD) is a chronic, progressive, neurodegenerative disease. People with PD are particularly susceptible to weight loss and malnutrition. Involuntary movements associated with PD result in increased energy expenditure, while both disease symptoms and medication side-effects can limit food intake. In addition, patients with the disease may choose to follow unconventional nutritional therapies that exacerbate malnutrition. Dietitians play a key role in helping patients with PD to optimize their nutritional status and manage various nutrition-related symptoms and medication side-effects. To assume this role, dietitians need to have current knowledge about PD and its nutritional consequences, as well as strategies for managing a variety of nutrition-related symptoms. PMID- 12084186 TI - Outcome after radical prostatectomy depends on surgical technique but not approach. PMID- 12084187 TI - Linkage disequilibrium between the androgen receptor gene CAG and GGC repeats in the African-American population. AB - African-American men are at an increased risk of developing prostate cancer when compared with other racial and ethnic groups. In addition, African-Americans display a greater propensity for developing aggressive prostate cancer. There are multiple etiologic factors that likely contribute to the development of prostate cancer; however, one potential factor that may explain differences of prostate cancer risk among ethnic and racial groups is the androgen receptor (AR) gene. Studies have showed that there is correlation between two polymorphic microsatellite regions of the AR receptor gene and its transactivational activity. The CAG and GGC repeats both have been implicated as important loci for variation in differential androgen receptor activity. This review analyzes the available data regarding variation of the CAG and GGC repeat sequences among different racial and ethnic populations, and the implications of these variations for prostate cancer risk. PMID- 12084188 TI - The role of transrectal ultrasound in diagnosing prostate cancer. AB - The availability of the prostate-specific antigen test to screen for prostate cancer has caused a significant workload burden for urologists and radiologists alike. Prostate cancer is common in our aging population and most cancers are now definitively detected by transrectal ultrasound (TRUS)-guided prostatic needle biopsy. TRUS alone has limited potential to identify prostatic cancer because of frequent multifocality of cancer within the prostate, the variable sonographic appearance of prostatic tumors, the poor specificity of focal ultrasonic abnormalities, and the substantial percentage of isoechoic prostate cancers (which cannot be differentiated from adjacent benign tissues with imaging). Developments in TRUS equipment over the past decade include the use of color and power Doppler, higher frequencies, broad bandwidth technologies, and harmonic, contrast harmonic, and pulse inversion imaging. All of these improvements may enhance detection of subtle focal sonographic abnormalities within the prostate. Ultrasonic contrast agents can aid the visualization of subtle alterations in prostatic echotexture by highlighting changes in microvasculature. It is possible that Doppler techniques and contrast agents have the potential to reveal prognostic information about cancer in individual patients. PMID- 12084189 TI - Permanent seed implantation for localized adenocarcinoma of the prostate. AB - Prostate brachytherapy has become a popular treatment option for localized prostate cancer with over 44,000 procedures performed in 2000. Eighty-seven percent to 93% of patients who have a serum prostate-specific antigen less than 10 ng/mL, Gleason score of 6 or less, and low risk (disease stage < or = T2a) can be expected to have an 8 to 10 year disease-free rate. The radiation dose delivered by the implants should exceed 140 Gy in men implanted with I-125 monotherapy. Patients with intermediate- and high-risk prostate cancer would benefit from the addition of either hormonal therapy and/or external beam irradiation to the implantation of seeds. Postimplant incontinence and proctitis can be minimized by controlling high radiation doses to the urethra and rectum. Potency is preserved in 70% of men with good preimplantation erectile function. Advances in technology, such as intraoperative dosimetry, will continue to make brachytherapy an attractive treatment option for men with localized prostate cancer. PMID- 12084190 TI - The effect of 15-lipoxygenase-1 expression on cancer cells. AB - Lipoxygenases oxidatively metabolize polyunsaturated fatty acids to a rich spectrum of biologically active products. One enzyme of the lipoxygenase family, Arachidonate 15-lipoxygenase-1 (arachidonate:oxygen 15-oxidoreductase, EC 1.13.11.33), is implicated in several forms of epithelial cell cancers, and provides a rationale for exploring its role in the progression and metastasis of cancers. Thus, 15-LO-1 could be a marker of the growth and spread of cancers. The importance of 15-LO-1 expression is imparted by its metabolic product of linoleic acid, yielding 13-(S)-hydroxyoctadecadienoic acid (13-(S)-HODE). In order to maximize our understanding of the roles of lipoxygenases and cyclooxygenases in cancer, it is necessary to correlate their metabolic pathways to the respective expression levels, and not exaggerate the presence or absence of an enzyme or its function when it clearly is not there. It is also important to acknowledge that some forms of these enzymes may not even be relevant to human disease. Although it is unlikely that any single gene is sufficient to account for the complex etiology of cancer, 15-LO-1 overexpression is clearly a key component in the cell proliferation process. This article is divided into three sections emphasizing the key role of 15-LO-1 in prostate, colorectal, and breast cancers. PMID- 12084191 TI - Diagnostic accuracy of prostate needle biopsy. AB - The introduction of prostate-specific antigen screening has resulted in stage migration and an increased incidence of localized prostate cancer. In this era of increasing nonpalpable disease, it has become necessary to systematically sample the entire prostate gland. Transrectal ultrasound-guided prostate biopsy procedures have evolved greatly over the past decade from the original sextant biopsy. Technological advances, better understanding of zonal anatomy of the prostate, whole mount sectioning of radical prostatectomy specimens, and computer modeling of localized prostate cancers have all led to extended biopsy core protocols directed at the lateral zones of the gland. These have increased the diagnostic accuracy of needle biopsy and have become a standard regimen. However, it remains controversial how to proceed with repeat biopsy in the face of an initial benign diagnosis, and optimal biopsy strategy remains undefined. It is hoped that quantitative analysis of prostate biopsy histology may eventually provide some prognostic information to guide the patient and urologist in preoperative planning. PMID- 12084193 TI - Hyperbaric oxygen for the treatment of radiation cystitis and proctitis. AB - Chronic radiation cystitis and radiation proctitis are known complications of the use of radiotherapy in the treatment of pelvic malignancies. These complications are, in part, due to endothelial damage and decreased vascularity and oxygenation to pelvic tissues. Hyperbaric oxygen therapy may be able to improve oxygenation and induce angiogenesis in damaged organs, resulting in recovery from radiation injury. Several studies have shown significant rates of response to hyperbaric oxygen treatment, however, no randomized trial exists to definitively demonstrate its effectiveness for cystitis and proctitis. In addition, concerns exist regarding the durability of the beneficial effect. PMID- 12084192 TI - Tumor hypoxia and the progression of prostate cancer. AB - Tumor cell hypoxia is an innate environmental factor encountered during the development of many types of human tumors, including malignant prostate tumors. For prostate cancer, however, tumor cell hypoxia may be an even more critical element in tumor development and progression. Recent evidence suggests that androgenic steroids are important regulators of blood flow to prostate tumors and suppressors of tumor cell hypoxia. In addition, because prostate tumor cells are similar to other eukaryotic cells, they have the ability to respond to hypoxic conditions with drastic changes in gene expression mediated by the upregulation of a unique transcription factor, hypoxia-inducible factor-1. This response increases cancer cells' metabolic resistance to hypoxia, and also enhances the ability of prostate cancer cells to attract a more vigorous blood supply by upregulating the expression of pro-angiogenic factors. Because such changes would, in essence, increase the potential aggressiveness of affected prostate cancer cells, it is clear that tumor hypoxia has the potential for being a very important factor in prostate cancer cell biology. This review focuses on recent studies regarding the occurrence and potential role of hypoxia in prostate cancer, including hypoxia-inducible factor-1 and its related signaling pathways. PMID- 12084195 TI - Quality-of-life differences among various populations of localized prostate cancer patients: 2001. AB - An increasing number of studies are being published on the impact of localized prostate cancer on patient quality of life. Research suggests that the majority of patients diagnosed with prostate cancer return to pretreatment levels of general quality of life within 1 year after treatment. However, sexual, urinary, and bowel symptoms continue to be an issue for many patients beyond 1 year post treatment. Focusing on psychologic impacts and social functioning, in addition to physical symptoms, may provide useful avenues for improving patient quality of life. We also discuss emerging evidence of racial and/or ethnic disparities in prostate cancer-related quality of life, the role of social networks and support in recovery and adjustment, as well as the impact of cancer recurrence. PMID- 12084194 TI - Effects of docetaxel on pain due to metastatic androgen-independent prostate cancer. AB - Bone pain commonly plagues patients with metastatic androgen-independent prostate cancer. Studies of mitoxantrone demonstrated that chemotherapy can substantially reduce this debilitating symptom. Two of the available studies examining the use of docetaxel with and without estramustine for treatment of androgen-independent prostate cancer include a detailed prospective analysis of pain and quality of life. One study required patients to have pain at entry and demonstrated significant improvement in pain. The second study enrolled patients with low prevalence and intensity of pain and did not demonstrate pain relief. The available results, although preliminary, suggest that patients with significant bone pain due to androgen-independent prostate cancer can experience substantial pain relief with docetaxel-based therapy. Larger randomized studies targeting patients with sufficient prevalence and intensity of pain are needed to refine our understanding of the contribution of docetaxel to pain control in this patient population. PMID- 12084196 TI - The use of bisphosphonates in the management of bone metastatic prostate cancer. AB - Improved understanding of the pathophysiology of prostate cancer-induced metabolic bone disease implies that bisphosphonates may have a role in the treatment of this disorder. Although prostate cancer cells stimulate osteoblasts, resulting in a marked increase in bone volume and the classic radiologic aspect of sclerotic metastases, accelerated bone resorption has been reported by histomorphometric and biochemical studies. The significant relationship between the biochemical parameters of bone resorption and formation suggests a coupling between these two processes in metastatic prostate cancer. Several studies have demonstrated that administration of bisphosphonates is one of the most cost effective palliative treatments for patients with prostate carcinoma and bone metastases, both as first-line and long-term therapy. With appropriate doses, a large proportion of patients can be maintained free of bone pain until death. Studies of the efficacy of lower doses to prevent skeletal morbidity in patients without metastases or with asymptomatic bone lesions are warranted. PMID- 12084197 TI - Brachytherapy versus radical prostatectomy in patients with clinically localized prostate cancer. AB - In an effort to help physicians offer their patients unbiased advice on the best alternatives for treatment of localized prostate cancer, we present a retrospective comparison of the effectiveness of brachytherapy and radical retropubic prostatectomy in 1305 men with stage T1 and T2 adenocarcinoma of the prostate. Data from 1305 patients treated in our community-based private practice urology group from 1993 to 2002 were reviewed, and patients were classified by initial prostate-specific antigen (PSA) level and risk grouping. Risk grouping was defined by preoperative PSA levels and Gleason scores. We used time to PSA indicated recurrence as the measure of efficacy. Brachytherapy and radical prostatectomy provided similar responses to treatment (no significant differences given the sample size, length of follow-up, and numerical differences observed) for localized prostate cancers. A prospective study is presently underway to evaluate the respective outcome of these procedures (including incidence of incontinence and impotence), and assess their impact on patient quality of life. The results presented here fail to show any superiority of prostatectomy over brachytherapy with palladium-103 (TheraSeed; Theregenics Corp., Buford, GA) with respect to time until relapse indicated by PSA level increase (> 0.2 ng/mL for prostatectomy and >1.5 ng/mL and rising for brachytherapy). In fact, any differences between treatments favor brachytherapy, particularly for intermediate and high-risk groups. We conclude that both brachytherapy and prostatectomy should be offered, equally and without bias, to men with stage T1 or T2 organ confined prostate cancer. PMID- 12084198 TI - Guidelines on benign prostatic hyperplasia: where do we stand in the new millennium? PMID- 12084200 TI - The adolescent varicocele: treatment and outcome. AB - At present, it is reasonable to counsel all patients, as well as family members of patients, who have a palpable varicocele about the long-term risks for impaired fertility. Currently, no one can predict with absolute certainty if any adolescent with a varicocele will be at risk for future infertility. Based on existing data, however, it is not reasonable simply to ignore the potential for such a problem until infertility becomes an issue because by that time the chance for reversibility may be lost. It is important to present a balanced discussion because the majority of men with a varicocele will be fertile, and no one is suggesting that all men with a varicocele undergo surgical treatment. Current recommendations for repair are based on the findings of impaired testicular growth and/or spermatogenesis. With early evaluation and selective treatment, however, we should be able to reduce the potential for future fertility problems significantly in adolescents with a varicocele. PMID- 12084201 TI - Vesicoureteral reflux: a new treatment algorithm. AB - Vesicoureteral reflux (VUR) affects about 1% of all children and carries an increased risk of pyelonephritis and long-term renal impairment. There are several approaches to the treatment of VUR: antibiotic prophylaxis (conservative treatment), open surgery, and endoscopic treatment. For many patients, endoscopic treatment cures VUR with a single procedure, eliminating the need for long-term antibiotic treatment and avoiding the trauma of a major surgical procedure. The choice of material for endoscopic treatment is of key importance, and, until recently, all available materials were associated with concerns regarding safety and efficacy. Emerging data demonstrate that dextranomer/hyaluronic acid (Dx/HA) copolymer has good long-term safety and efficacy in treating VUR. A new treatment algorithm is, therefore, proposed, recommending that most children with persistent VUR (longer than 1 year) be offered endoscopic treatment with Dx/HA copolymer as an alternative to prolonged antibiotic prophylaxis or open surgery. PMID- 12084202 TI - The valve bladder: etiology and outcome. AB - The infravesical obstruction created by posterior urethral valves produces a series of structural, biochemical, and functional changes in these boys' bladders that begins during fetal life. Any change can be irreversible and result in the alterations in bladder function that can be found in some of these patients. Bladder instability, poor compliance, and myogenic failure are the three most common urodynamic patterns and are also responsible for some of these patients' poor long-term prognosis. Treatment of bladder dysfunction in these patients is basically directed toward improving their kidney function prognosis and also to avoid the urinary incontinence presented secondarily by some valve bladders. PMID- 12084203 TI - The endoscopic treatment of incontinence in children. AB - The achievement of urinary continence in children may be very difficult with some congenital malformations of the lower urinary tract, such as bladder exstrophy, or in some neurogenic dysfunctions, such as neuropathic bladder. Endoscopic injection of bulging substances into the bladder neck or posterior urethra may be regarded as a minimally invasive procedure that may provide a better dry interval in these patients. Endoscopic treatment can be performed as either a primary or adjunctive procedure after open bladder neck surgery. It is also of use to increase the functional bladder capacity in some children. Although longer follow up is wise, results so far are encouraging. The search for a safe and reliable substance to use is still ongoing, and the new available biodegradable implants seem adequate. PMID- 12084204 TI - Advances in bladder augmentation. AB - Over the past 20 years, rapid advances have been made in bladder augmentation due to the introduction of clean intermittent catheterization, the use of intestinal segments interposed into the urinary tract, and the development of tissue expansion. The particular augmentation method selected from the multiple methods currently available is an individualized process that takes into account both patient factors and potential complications. Exciting new techniques, such as tissue engineering, may change the face of lower urinary tract reconstruction. PMID- 12084205 TI - Antenatal hydronephrosis: evaluation and outcome. AB - We present a comprehensive and current review of the etiology, evaluation, treatment, and outcome of antenatal hydronephrosis. When a diagnosis of antenatal hydronephrosis is made, many questions regarding pregnancy, prenatal care, intervention, and what may be expected after birth are raised. Debate and controversy exist on the diagnosis and subsequent evaluation and management of the child with antenatal hydronephrosis. A review of the literature and our own experience with antenatal hydronephrosis are presented in order to guide the physician who is caring for mother and child. PMID- 12084206 TI - Laparoscopic radical prostatectomy. AB - Laparoscopy has become an integral part of urologic surgery. Its indications have been progressively extended to the most advanced oncologic and reconstructive procedures. Within this frame, radical prostatectomy is of major interest, especially considering the incidence and clinical significance of prostate cancer. The procedure comprises several steps of challenging dissection in which the preservation of delicate nerve and muscular structures should be conciliated with safe tumor excision. The intervention ends with vesicourethral anastomosis, which is considered the most difficult reconstructive procedure in urologic laparoscopy. Laparoscopic radical prostatectomy has gradually become a wholly standardized procedure, and it is now routinely performed in several centers throughout the world. PMID- 12084207 TI - Comparison of laparoscopic radical prostatectomy techniques. AB - Over the past years, laparoscopic radical prostatectomy has emerged as an attractive, exciting, and new approach for the surgical treatment of localized prostate cancer. Several centers in Europe and the United States have developed their own technique and have already published their first results. Two main routes have been used, namely, the transperitoneal and extraperitoneal approaches. Data from the literature and available experience demonstrate that this laparoscopic procedure is feasible and teachable and that oncologic and functional results seem comparable to those of classic open radical prostatectomy. Minimal bleeding, reduced blood transfusion rates, shorter hospitalization, and shorter recovery time are unquestionable advantages for laparoscopic procedures. Laparoscopic radical prostatectomy remains a difficult intervention and should be performed in selected centers with experienced teams who already have extensive laparoscopic experience and who recruit enough patients to grant them the proper amount of expertise. This is the only way to achieve excellence in terms of oncologic and functional results. PMID- 12084209 TI - Japanese experience with radical prostatectomy. AB - Laparoscopic prostatectomy has become an established treatment option for localized prostate cancer in France, where open prostatectomy is now the gold standard. The main purposes of treatment for prostate cancer are cancer control and preservation of urinary continence and sexual function. To become a standard treatment option for organ-confined prostate cancer, laparoscopic prostatectomy has to show equal or better clinical outcome in these areas than its open counterpart. Many institutes in other countries are now trying to perform this surgery. There are, however, some negative reports, mainly because of the difficulty of the procedure. In Japan, more than 250 patients have undergone this surgery. It seems that satisfactory results in terms of positive surgical margin rate, bleeding volume, recovery from surgery, and urinary continence have been obtained so far. We need longer follow-up to assess recurrence rate and sexual function. The main obstacles for this surgery are the long operative time and the difficulty of the procedures. Although it will take time until laparoscopic prostatectomy becomes an approved treatment modality, we are gradually conquering these problems. In this paper, we review the current situation facing laparoscopic prostatectomy in Japan. PMID- 12084208 TI - Laparoscopic radical prostatectomy: published series. AB - Next to the retropubic and perineal approaches, laparoscopic radical prostatectomy has become the third most common technique in the surgical treatment of localized prostate cancer. Although long-term oncologic data are still lacking, based on several contemporary series, it seems likely that oncologic results will fulfill expectations. Over the past decade, quality-of life issues have come into the spotlight in oncologic surgery in particular. In this regard, the aim of the laparoscopic technique is to become the best in terms of operative stress, postoperative morbidity, catheterization time, and return to normal activities. The unique intraoperative visibility and magnification of the operative field allow ultraprecise dissection and suturing of vital neural and sphincteric structures. Will the use of this new kind of surgery translate into better functional results? The goal of this review is to analyze the published results of laparoscopic radical prostatectomy in the context of its rapidly evolving open surgical counterpart. PMID- 12084210 TI - Laparoscopic radical nephrectomy: retroperitoneal versus transperitoneal approach. AB - Laparoscopic radical nephrectomy can be efficaciously performed by either the transperitoneal or the retroperitoneal laparoscopic approach. The primary indication for selecting one approach over another has historically depended on the individual surgeon's experience and training. With either technique, laparoscopy adheres to established surgical oncologic principles of wide specimen mobilization and early vascular control. This article reviews the history, contraindications, anatomic considerations, patient preparation, and surgical technique of these two laparoscopic approaches. A salient summary of the worldwide experience with these procedures is presented, as well as a brief synopsis of controversial arguments favoring specimen morcellation versus intact extraction. PMID- 12084211 TI - Laparoscopic adrenalectomy: alternative or new standard? AB - Laparoscopy has become a standard approach for adrenalectomy because of its safety, low invasiveness, and less demanding technical nature and the readily removable size of tumor through trocar incision. Comparative studies between open and laparoscopic adrenalectomy document less blood loss, shorter hospital stay, and lower incidence of complication. These reports also show that the patients have less pain, use fewer narcotics postoperatively, and have quicker resumption of oral intake after surgery with the laparoscopic approach. The techniques for laparoscopic adrenalectomy started with the transperitoneal approach and developed into the retroperitoneal approach. Further technical development and recognition yielded three transperitoneal and two retroperitoneal approaches. Characteristics of each approach are discussed. Due to technical developments and experiences in laparoscopic surgery, application of the laparoscopic approach has been expanded to include excision for adrenal cancer and laparoscopic partial adrenalectomy for bilateral pheochromocytoma in certain cases and in selected institutes. PMID- 12084212 TI - Prostate imaging in malignant and benign conditions. PMID- 12084213 TI - A randomized phase III trial of high dose interleukin-2 versus subcutaneous interleukin-2/interferon. PMID- 12084215 TI - Laparoscopic radical nephrectomy for advanced kidney cancer. AB - The management of advanced renal cell carcinoma (RCC) continues to evolve. With the advent of laparoscopic radical nephrectomy (LRN), minimally invasive approaches to kidney cancer have developed. Laparoscopic resection of locally advanced RCC yields a similar cancer-control rate with the advantage of decreased morbidity. Although cytoreductive LRN is a technically challenging procedure, it may be completed safely in selected patients. Further prospective study of the role of LRN for advanced RCC is warranted. PMID- 12084214 TI - Minimally invasive ablative approaches in the treatment of renal cell carcinoma. AB - Ablative techniques for the treatment of renal cell carcinoma (RCC) are an extension of nephron-sparing surgery and include cryoablation, radiofrequency ablation (RFA), and high-intensity focused ultrasound (HIFU). Although experimental, these are evolving treatment modalities. The widespread use of computed tomography scans, ultrasound, and magnetic resonance imaging caused an increase in the diagnosis rate of small renal masses amenable to nephron-sparing surgery. The same imaging modalities permit interactive monitoring both during the delivery of ablative measures and at postoperative follow-up. Cryosurgery is the most studied of the ablative approaches, and clinical studies have demonstrated promising short-term results and a remarkable safety profile. Long term studies, however, are needed in order to determine the appropriate selection criteria and to confirm a response as durable as that for partial and radical nephrectomy. More data are needed to evaluate the efficacy of RFA. Currently, preclinical results with HIFU do not justify its use for treating RCC in humans. PMID- 12084216 TI - Kidney pathology: current classification of renal cell carcinoma. AB - This report discusses the new cytogenetic classification of renal cell carcinoma (RCC) and its biological and clinical significance. It describes the four major types (clear cell, chromophil, chromophobe, and collecting duct) as well as rarer entities, such as small cell carcinoma, cystic RCC, rhabdoid variant of RCC, and unclassified RCC. Sarcomatoid carcinomas are a diverse group representing high grade transformation of the other types. In addition to a description of the pathologic findings, there is a discussion of tumor markers and prognostic indicators. The clinical significance of the classification is discussed, with reference to recent reports. PMID- 12084218 TI - Surgical management of renal cell carcinoma invading the vena cava. AB - Surgery remains the most effective treatment for renal cell carcinoma (RCC) invading the vena cava. Removal of tumor involving the vena cava or atrium may be challenging, but a thorough preoperative assessment and careful surgical technique have enhanced safety of resection and improved outcome. Preoperative imaging is essential for effective surgical intervention. Tumor extension into the vena cava does not adversely influence outcome in most reports. Characteristics of the primary tumor, including grade, perinephric fat invasion, lymph node involvement, and metastases, dictate outcome. Metastases are not an absolute contradiction to surgery, given the encouraging results with adjuvant immunotherapy. Nonetheless, improved methods for appropriate patient selection and new adjuvant therapies are needed to complement surgical intervention. PMID- 12084217 TI - Prognostic factors for biologic therapy in kidney cancer. AB - Renal cell cancer is resistant to most forms of therapy. Cytokine therapy with either interleukin-2 or interferon-alpha yields the best results, with response rates from 10% to 20%. Therapy is not without toxicity, which means that the majority of patients treated with cytokines suffer toxicity without any therapeutic benefit. Recent endeavors have tried to find new ways to identify responders to cytokine therapy. Prognostic factors, such as good performance status, lack of anemia, normal calcium, normal lactate dehydrogenase, and prior nephrectomy, correlate with an increased likelihood of responding to cytokine therapy. Recent studies have examined whether altered subpopulations of lymphocytes, the presence of eosinophils, or altered levels of cytokines can predict response to cytokine therapy. Although prior nephrectomy does correlate with improved survival while the patient is receiving cytokine therapy, it is unclear if this is due to a significant alteration in the tumor's response to cytokine or some unrelated benefit from surgery. Further studies are needed to confirm the current immune parameters and disease characteristics that suggest a better response to cytokine therapy. PMID- 12084219 TI - The role of adjuvant immunotherapy in renal cell carcinoma. AB - After nephrectomy for renal cell carcinoma (RCC), a significant number of patients develop recurrent disease. In order to improve the prognosis of these patients, the role of adjuvant immunotherapy should be clarified; the appropriate selection of patients is especially crucial. For this purpose, the search for prognostic factors is important to identify at-risk patients. Known factors such as stage, grade, and microvascular invasion can be used for appropriate selection. Other molecular markers, such as cadherin-6 and G250 antigen, may become important. So far, adjuvant immunotherapy in RCC has not shown improved survival data, but the results may be hampered by inadequate recruitment and follow-up. Adequate selection of patients and the search for less toxic and more effective immunotherapy approaches are of importance. Therefore, the use of monoclonal antibody G250 or dendritic cell vaccinations, alone or together with cytokines, may be advantageous and is currently used. Today, adjuvant protocols are open for recruitment of patients to elucidate the important question as to whether this approach should be implemented in the treatment of RCC. In this article, an update is given in the field of adjuvant immunotherapy in patients with RCC. PMID- 12084220 TI - DNA microarrays in prostate cancer. AB - DNA microarray technology provides a means to examine large numbers of molecular changes related to a biological process in a high throughput manner. This review discusses plausible utilities of this technology in prostate cancer research, including definition of prostate cancer predisposition, global profiling of gene expression patterns associated with cancer initiation and progression, identification of new diagnostic and prognostic markers, and discovery of novel patient classification schemes. The technology, at present, has only been explored in a limited fashion in prostate cancer research. Some hurdles to be overcome are the high cost of the technology, insufficient sample size and repeated experiments, and the inadequate use of bioinformatics. With the completion of the Human Genome Project and the advance of several highly complementary technologies, such as laser capture microdissection, unbiased RNA amplification, customized functional arrays (eg, single-nucleotide polymorphism chips), and amenable bioinformatics software, this technology will become widely used by investigators in the field. The large amount of novel, unbiased hypotheses and insights generated by this technology is expected to have a significant impact on the diagnosis, treatment, and prevention of prostate cancer. Finally, this review emphasizes existing, but currently underutilized, data-mining tools, such as multivariate statistical analyses, neural networking, and machine learning techniques, to stimulate wider usage. PMID- 12084221 TI - Sacral neuromodulation for the treatment of bladder dysfunction. AB - S3 nerve root neuromodulation is becoming an accepted therapy for individuals afflicted with lower urinary tract symptoms, such as idiopathic urinary urge incontinence, frequency, urgency, and urinary retention, who fail current standard therapies. This patient population is difficult to treat, and this therapy offers an option that can restore quality of life to these individuals. While the precise pathophysiologic mechanisms underlying these voiding symptoms is unknown based on the physiology of bladder function, it is logical to conclude that they relate to some aberration of the neuromuscular apparatus of the bladder and/or its outlet (the urethra). These pathophysiologic defects are not overtly manifested outside the lower urinary tract because these patients routinely do not have any apparent neurologic deficits. The fact that S3 neuromodulation can clinically improve these lower urinary tract symptoms would support the notion that the neural regulation of the bladder is somehow altered in these patients. This paper presents a review of the current knowledge regarding sacral neuromodulation in treatment of non-neurogenic voiding dysfunction. PMID- 12084222 TI - A novel method for the analysis of the androgen receptor. AB - The role of the androgen receptor in the regulation of prostate cancer development and progression has been a focus of intense research. Until recent years, the level of expression of the androgen receptor protein was described qualitatively. Immunohistochemical parameters have been established that show a linear relationship between androgen receptor expression and immunostaining. Intensity of immunostaining can be accurately measured using computer-assisted color video image analysis. Studies of progression and treatment of prostate cancer will benefit from the ability to quantitatively measure androgen receptor expression. PMID- 12084223 TI - Gene therapy for urologic cancer. AB - Advances in molecular technology and the completion of the human genome project have ushered in a new era of medicine, that of gene therapy. In every field of medicine, investigators are developing gene therapeutics in an attempt to cure diseases. Urologic oncology is no exception. Herein, we review the current status of gene therapy for urologic malignancy. Included is an overview of advances in gene delivery systems and immunology, which are driving forces for gene therapy research. Finally, we review the current gene therapy trials and experimental approaches for urologic malignancy. PMID- 12084225 TI - Human comfort: more than modern medical technology. PMID- 12084224 TI - Immune gene therapy in urology. AB - Effective treatments are needed urgently for metastatic disease in bladder, prostate, and renal cell cancer. In the past few years, several new approaches for treating these conditions have been proposed, including gene therapy. A number of different strategies have been developed to accomplish urologic cancer gene therapy. Genetic immunomodulation strategies attempt to activate immune defense mechanisms against tumor cells by transfer of tumor antigens, cytokine genes, or strongly immunogenic cell surface molecules. In this review, we illustrate the recent developments in immune gene therapy. PMID- 12084227 TI - The pathophysiology of varicocele-associated male infertility. AB - Varicocele is the most commonly identifiable, surgically correctable lesion associated with male-factor infertility. Surgical correction of a varicocele, whether unilateral or bilateral, results in improvement not only in semen parameters but also in spontaneous and assisted pregnancy rates. Varicoceles seem to induce a number of changes in the testicular microenvironment. These alterations in temperature, hemodynamics, and reactive oxidative species and antioxidant concentrations have been demonstrated to produce deleterious effects on spermatogenesis. However, despite current knowledge in the pathophysiology of varicocele-associated male infertility, the exact mechanism--or mechanisms--by which varicoceles impair fertility remains elusive. This review examines scientific evidence regarding the pathophysiology of varicocele-associated male infertility. PMID- 12084226 TI - Is intracytoplasmic sperm injection safe? Current status and future concerns. AB - In vitro fertilization with intracytoplasmic sperm injection (ICSI) has revolutionized the treatment of patients with severe forms of male infertility. However, because ICSI has been performed for only 10 years, long-term outcomes and risks to offspring remain largely unknown. The fact that ICSI can potentially bypass natural selection barriers to genetic disease transmission has brought a sobering but important impetus to recent research on the risks and outcomes of ICSI. Several studies were done recently to examine specific risks to children born following ICSI. Because of rapid advances in the ICSI procedure itself, studies evaluating the safety of using immature sperm forms from the testis (spermatids, spermatocytes) also have been undertaken. This review summarizes recent studies examining the risks and long-term outcomes to date of in vitro fertilization with ICSI. PMID- 12084228 TI - Ultrasound and infertility: diagnostic and therapeutic uses. AB - Ultrasound has become essential in the diagnosis and management of the infertile male. Scrotal ultrasonography provides a detailed examination of the testes and assesses the presence or absence of varicoceles and can identify other abnormalities of the scrotal contents and the spermatic cord. Transrectal ultrasonography can "visualize" the excurrent ejaculatory ductal system, including the ejaculatory ducts, seminal vesicles, and vas deferens. The expansion of these ultrasonographic techniques has provided the urologist with non- or minimally invasive techniques with which to evaluate the infertile male. These advancements consequently have led to innovative surgical and radiologic treatments. PMID- 12084229 TI - Predictors of success in microsurgical correction of vasal and epididymal obstruction. AB - Patients who inquire about microsurgical correction of vasal and epididymal obstructions should be informed about the postoperative rates of both return of sperm to the semen and pregnancy. They also should be informed about the possibility of a successful outcome, depending on their individual circumstances. The various preoperative, intraoperative, and postoperative factors that influence success rates are discussed in this article. PMID- 12084230 TI - Diagnosis and treatment of the azoospermic patient. AB - With the advent of intracytoplasmic injection, the management of azoospermia has become ever more important. Gametic manipulation to produce biological offspring is not feasible unless sperm from the azoospermic male patient is obtainable. This article provides an overview of the diagnosis and treatment of azoospermia. PMID- 12084231 TI - Androgen deficiency and the aging male: new urologic perspectives. AB - Among the most prominent health care issues of the 21st century are those relating to aging. Indeed, the elderly represent the fastest growing segment of our population. Typically, a urology practice includes a significant number of elderly males, providing the physician with both a challenge and an opportunity. Androgen deficiency in the aging male, with its detrimental effects on bone, muscle and body composition, sexual function, and psychological well-being, is an area of increasing interest in both the lay press and the medical community. The urologist must understand the pathophysiology of androgen deficiency in the male, recognize and investigate the subtle signs and symptoms of hypogonadism, determine the appropriateness of treatment, and offer informed options to the patient. Health care professionals have a responsibility to make "healthy" aging a priority. PMID- 12084232 TI - Sexual dysfunction and depression: etiology, prevalence, and treatment. AB - Sexual dysfunction and depression are very common conditions that are age-related and chronic. In men, epidemiologic studies have confirmed a strong correlation between erectile dysfunction and symptoms of depression. Both conditions have a significant negative impact on the quality of life of patients and their partners. Several studies showed that restoration of normal sexual function improves the quality of life of patients and their partners, regardless of treatment method. The literature review and recent observations emphasize the multifactorial nature of sexual dysfunction and, more specifically, erectile dysfunction and underline the importance of the comorbidity and bidirectional relationship between erectile dysfunction and depression. Research is progressing on the possible link between andropause, sexual dysfunction, and depression, thus opening potential new opportunities to address issues of aging-related morbidities. PMID- 12084233 TI - Intracavernosal therapy: when oral agents fail. AB - Erectile dysfunction is a common problem in men older than 40 years, and the incidence increases markedly with age. Approximately one fourth of impotent men use intracavernosal injections as a primary treatment. Since its original description in the early 1980s, injection therapy has been used safely and successfully by many men and their partners. Intraurethral and oral agents for the treatment of erectile dysfunction have been introduced over the last few years, but early reports of patient satisfaction and reproducibility of erectile rigidity have not matched those of intracavernosal therapy. Thus, injection therapy remains on the forefront of treatment options. PMID- 12084234 TI - Androgen deficiency in the aging male: pathophysiology, diagnosis, and treatment alternatives. AB - Androgen deficiency in the aging male is associated with negative effects on mood and cognition, diminished sexual function, and osteoporosis. The aging of our population, combined with an increased acceptance of medical technology, has focused greater attention on optimizing health during the "golden years." Androgen replacement therapy may remedy many of the deleterious consequences of hypogonadism. This communication reviews the clinical manifestations and diagnosis of androgen deficiency, the inherent risks and benefits of androgen replacement, and the options of testosterone replacement currently available. PMID- 12084235 TI - Gene therapy: future therapy for erectile dysfunction. AB - Advances in molecular biological techniques, completion of the Human Genome Project, and the ensuing age of molecular medicine, in conjuction with the sum of a decades-long accumulation of knowledge of the physiology of erection and the pathophysiology of erectile dysfunction have converged to make gene therapy for erectile dysfunction a distinct possibility. In short, both the intrinsic complexities of mechanisms responsible for ensuring normal erection and the multifactorial nature of erectile dysfunction ensure that there is a relatively vast number of physiologically relevant molecular targets for gene therapy. As such, perhaps it is not surprising that virtually every preclinical gene therapy strategy/target examined thus far has been largely successful in ameliorating conditions associated with compromised erectile function in vivo and/or in vitro. This report highlights the goals and strategies of gene therapy for erectile dysfunction and reviews the strategies that initially have been employed. In short, the preclinical data, while still quite preliminary in many regards, are nonetheless quite impressive and encouraging. If similar success is obtained in clinical trials, gene therapy for erectile dysfunction may provide the first concrete "proof of concept" for using gene therapy in the treatment of human smooth muscle disorders. PMID- 12084236 TI - Central nervous system agents in the treatment of erectile dysfunction: how do they work? AB - Drugs acting within the central nervous system (CNS) that reduce the sympathetic antierectile flow and enhance the parasympathetic proerectile flow to the penis may restore penile erection in cases of erectile dysfunction of both psychogenic and organic origin. The best characterized of such drugs is the dopaminergic agonist apomorphine, which acts on the hypothalamus and, perhaps, the autonomic nuclei in the spinal cord. Other drugs that target the CNS and have been registered and tested are the a(2)-adrenoceptor antagonists yohimbine and delequamine, the alpha-melanocyte-stimulating hormone agonist melanotan II, and the serotonin reuptake inhibitor trazodone. Androgens also may influence sexual behavior by acting within the CNS, notably by modifying the neurotransmitter system targeted by these drugs. Our knowledge of the mode of action of CNS drugs comes mainly from experiments on rodents. Consequently, explanations regarding the way they work in humans are only speculative. PMID- 12084238 TI - What's my scale? Organ injury severity scaling in the new millennium. PMID- 12084239 TI - Surgery or collagen for the treatment of female stress urinary incontinence: results of a multicenter, randomized trial supports either as first line of treatment. PMID- 12084237 TI - Management of erectile dysfunction following radical prostatectomy. AB - Radical prostatectomy is the standard treatment for organ/specimen-confined prostate cancer, yet erectile dysfunction in selected series is still reported as high as 90% after this procedure. Thus, most men need adjuvant treatments to be sexually active following radical prostatectomy. These include vacuum constriction devices, intracorporeal injections of vasoactive drugs, and transurethral dilators, all of which have reported response rates of 50% to 70%. Unfortunately, long-term compliance is suboptimal, with a discontinuation rate of nearly 50% at one year. These non-oral options should be offered on an individual basis to patients who have failed oral therapy since efficacy and compliance vary. Also, these options should be considered in the early postoperative period to enhance sexual activity and penile oxygenation, which may prevent corporeal fibrosis. Early penile rehabilitation with intracavernosal injections or vacuum constriction devices should be encouraged to increase chances for recovery of rigid erections. In patients with some preservation of nerve tissue, oral sildenafil may be effective in promoting an earlier return of erectile function. The potential impact of sildenafil and other new oral therapies should encourage urologists to continue to perform and perfect the nerve-sparing approach. PMID- 12084240 TI - Transvaginal bladder neck suspension to Cooper's ligament. AB - Numerous surgeries have been proposed for the treatment of genuine stress incontinence, with the goals to improve functional outcome and decrease complications and their associated morbidity. Two new, minimally invasive procedures, transvaginal retropubic urethropexy and transvaginal Cooper's ligament sling, are reviewed in this article. These procedures provide a completely transvaginal approach, without the use of abdominal incisions or bone anchors. The anterior point of suspension is Cooper's ligament. The transvaginal retropubic urethropexy is used for the treatment of genuine stress incontinence with urethral hypermobility, and the transvaginal sling also may be used in the presence of intrinsic sphincteric deficiency. The procedures are described and the recent outcomes discussed. PMID- 12084242 TI - Nonautologous sling materials. AB - This article provides a comprehensive and updated review of the current nonautologous sling materials available for treatment of stress urinary incontinence. The various materials are described, relevant clinical studies are discussed, and newer materials are outlined. Complications arising from use of different materials also are described. PMID- 12084241 TI - Bladder substitution in women. AB - Bladder substitution in women with transitional cell carcinoma (TCC) is not a universally accepted procedure. There are many concerns, one of which is the potential risk of metachronous and synchronous urethral transitional cell carcinoma involvement. Another concern is that voiding dysfunction may be more frequent than in male patients. The numbers of female patients who have had this procedure are still small, and follow-up data are relatively brief. Thus, the true role for orthotopic bladder substitution in the female is still being evaluated. This paper reviews the data on this type of surgery in women, with emphasis on urethral TCC risk and on voiding dysfunction. Early results from a number of institutions are encouraging, particularly in correctly selected patients. Based on more than 15 years experience in an albeit small number of patients, we believe that if there is a functional external sphincter and tumor margins can be safely cleared, this form of surgery offers patients the best opportunity to preserve quality of life following cystectomy. PMID- 12084243 TI - A review of the tension-free vaginal tape procedure: outcomes, complications, and theories. AB - In the last century, the pathophysiology of stress urinary incontinence (SUI) has been investigated and several surgical techniques have been utilized for cure. The most recent evolution in the study of SUI is the minimally invasive tension free vaginal tape (TVT) procedure, which can be done under local anesthesia and intravenous sedation and is individualized for each patient. The procedure recreates the "hammock" of the anterior vaginal wall and endopelvic fascia with a mesh tape of polypropylene. The cure rates of the initial studies are equal to or better than other anti-incontinence procedures, and the permanent supportive mesh is very well tolerated. The TVT creates a backboard on which the urethra compresses itself when it rotates posteriorly during cough or stress. The procedure accomplishes subjective and objective cure without elevating the bladder neck or altering urethral mobility. PMID- 12084244 TI - Urethral diverticula: evolving diagnostics and improved surgical management. AB - Urethral diverticula remain problematic from both diagnostic and therapeutic standpoints. Recent developments in pelvic imaging with computed tomography, sensitive ultrasonography, and magnetic resonance imaging have greatly advanced diagnostic acumen and improved the clinician's ability to stage lesions as to location, size, and coexistent pathology. Coupled with improved recognition has come advancements in surgical technique, reflective of improved understanding of urethral anatomy and function. Better use of concomitant procedures, such as pubovaginal sling or soft tissue interposition, has continued a steady trend toward improved surgical outcomes when considering urethral function and urinary continence. This article reviews these mutually complementary trends. PMID- 12084245 TI - Bladder outlet obstruction in women: prevalence, recognition, and management. AB - Bladder outlet obstruction in women is an infrequently diagnosed urological condition. Its prevalence has been estimated to be between 2.7% and 29%. The large variation in prevalence is likely a result of the lack of standard diagnostic definition for the evaluation of female bladder outlet obstruction. A combination of history taking; physical examination; and diagnostic tests, including simple pressure void studies, radiographic imaging, endoscopic visualization, and multichannel videourodynamics provides a consistent way to accurately recognize and diagnose bladder outlet obstruction. Causes of obstruction are varied and numerous but generally fall within two broad categories: functional and anatomic. Treatment options are tailored to individual causes of obstruction and range from conservative pharmacologic and behavioral options to more-invasive surgical procedures. PMID- 12084246 TI - Valsalva leak point pressure: steps toward standardization. AB - Valsalva leak point pressure (VLPP) has been used as the urodynamic indicator of intrinsic sphincteric deficiency in patients with stress urinary incontinence. However, further validation of VLPP as a tool for diagnosis and assessment of treatment outcome has been delayed mainly because of the lack of a universally accepted standardized methodology. The urodynamic parameters in need of standardization for measurement of VLPP include urethral catheter size, zeroing of the transducer, patient position, bladder volume, type of stress, and timing of measurement. Such standardization likely will allow for further validation of VLPP and its use in distinguishing the right treatment options for the correction of stress urinary incontinence. PMID- 12084247 TI - Vaginal repair of large cystoceles. AB - Repair of cystoceles requires a complete understanding of the pelvic anatomy. While smaller defects are relatively straightforward, greater degrees of prolapse can be among the most challenging surgeries in pelvic floor reconstruction. This article reviews current transvaginal techniques used for repair of large cystoceles. PMID- 12084248 TI - New millennium, new slings. AB - The etiology of stress incontinence is not completely understood. In the past, bladder neck suspensions were performed to correct anatomic abnormalities of the bladder neck and urethral hypermobility. This procedure was attractive because of its simplicity, low morbidity, and excellent early success rate. With time, the successes seen with bladder neck suspensions have not proven to be durable, and alternative surgical procedures have been developed. Until recently, the indications for bladder neck suspension were types I and II stress incontinence; slings were reserved for type III incontinence. However, slings have been shown to be as effective as and more durable than bladder neck suspensions for treatment of all types of stress incontinence; therefore, their popularity has spread. The success of distal urethral slings suggests that it is not necessary to correct anatomic hypermobility to correct stress incontinence. A plethora of new procedures and materials has emerged, leading to an increased need for well controlled, objective outcome studies in order to understand the impact of these surgeries on our patients. PMID- 12084249 TI - Practical outcome measures for assessing efficacy of incontinence procedures. AB - Standard treatment for urinary incontinence in women has evolved during the past few decades. Conservative measures such as pelvic floor exercises and biofeedback may be effective and have been advocated for the past several years. However, the availability of other methods that are potentially more invasive yet efficacious provides a wider range of choices for women with urinary incontinence. With these alternatives comes opportunity and responsibility to assess how successful these treatments are. This article explores current methods of analyzing outcomes of urinary incontinence treatments. PMID- 12084250 TI - Surgery of the adrenal gland. PMID- 12084251 TI - Holmium laser prostatectomy. AB - Holmium laser prostatectomy has evolved in the last decade to reproduce the short and long-term results of both transurethral and open prostatectomy. This article discusses our surgical approach at the Methodist Hospital of Indiana as well as offers a review of the literature of holmium prostatectomy. PMID- 12084252 TI - Use of prostatic stents for the treatment of benign prostatic hyperplasia in high risk patients. AB - Benign prostatic hyperplasia (BPH) is a frequent disease in men and a major cause of lower urinary tract symptoms (LUTS). Transurethral resection of the prostate (TURP) or open surgery remains the gold standard of treatment for symptomatic BPH. However, 10% to 15% of patients with BPH cannot undergo surgery due to grave concomitant diseases. For patients presenting with contraindications to surgery or anesthesia, several minimally invasive alternative treatment modalities are available. One such therapeutic alternative is prostatic stenting, which can serve as a temporary or permanent solution for bladder outlet obstruction caused by BPH. Although not a new concept, this is a relatively new treatment modality in the United States, primarily because of the strict regulatory forces governing the use of these devices. Prostatic urethral stents have been widely demonstrated to be safe and effective for the treatment of symptomatic BPH. In addition to being minimally invasive, prostatic stenting is generally rapid, easy to perform, immediately effective, and has a low cost compared with conventional surgical treatment. Prostatic stents are therefore well suited to treat the frail elderly patient who would not be able to withstand the stress of undergoing surgery. This report reviews the current use of prostatic urethral stents in the treatment of high-risk surgical patients with BPH. PMID- 12084253 TI - Hormonal treatment of patients with benign prostatic hyperplasia: pros and cons. AB - The recognition that dihydrotestosterone is a "major player" in the development of benign prostatic hyperplasia (BPH) provided an impetus for the development of a 5a-reductase inhibitor, finasteride. During the past 5 years, a number of publications have noted that alpha blockers appear more efficacious than finasteride. This article reviews the role of hormones (particularly finasteride) in the treatment of lower urinary tract symptoms caused by BPH. These observations indicate that finasteride has a role in the management of larger prostates. Long-term use reduces the development of urinary retention and need for invasive procedures such as transurethral prostatectomy. The major adverse impact of finasteride is its effect on the patient's libido and sexual function. This is a less morbid problem for the elderly than the potential syncope associated with the use of alpha blockers. A greater understanding of the interaction of hormones on prostate receptors will provide newer tools for the treatment of BPH. PMID- 12084255 TI - Effect of diabetes mellitus on lower urinary tract symptoms and dysfunction in patients with benign prostatic hyperplasia. AB - This article reviews the recent literature on men with diabetes mellitus (DM) and concomitant lower urinary tract symptoms (LUTS) suggestive of benign prostatic hyperplasia (BPH). Diabetes is reported to be associated with greater BPH symptom severity. Men with non-insulin-dependent diabetes had the highest median annual prostate growth rate compared with those who had other metabolic disease. Urodynamic evaluations in the reviewed studies were absent. Extensive diagnostic evaluations with urodynamics in our outpatient department revealed that patients with LUTS suggestive of BPH and with concomitant DM hardly differed from those without DM. PMID- 12084254 TI - Sexual function in patients with benign prostatic hyperplasia. AB - Sexual health has significant impact on quality of life among men with benign prostatic hyperplasia (BPH). The degree of sexual dysfunction matches the severity of lower urinary tract symptoms (LUTS). Treatment of BPH affects not only LUTS, but sexual function as well. Medical, surgical, and minimally invasive therapies differ in their effect on erectile function, ejaculation, and sexual satisfaction. Choice of treatment modality takes into account baseline sexual function and patient expectations. This review outlines the relationship between LUTS and sexual function and how they change with the currently available treatments. PMID- 12084256 TI - Transurethral hot water balloon thermoablation. AB - The inclusion of a determinate technique among the "minimally invasive treatments" of the symptoms due to benign prostatic hyperplasia is a long and careful process. The technique has to be more patient-friendly than conventional techniques, but also needs to be time-tested against older methods. Most of these minimally invasive treatments are based upon distinct types of thermoablation; the method discussed here, hot water balloon thermoablation, is based on the heat conductive properties of the prostatic tissue. PMID- 12084257 TI - Lasers for median lobe hyperplasia. AB - Laser treatment encompases a variety of techniques using different laser wavelengths, application systems, and surgical techniques to achieve contrasting tissue effects such as incision, resection, vaporization, or coagulation. Many studies have proven the clinical efficacy of the various laser techniques for the treatment of benign prostatiuc hyperplasia, including randomized studies versus transurethral prostatectomy (TURP). Recently, long-term follow-up of up to 5 years has demonstrated the durability of the results, although in some of the studies, retreatment rates were higher than after TURP. Median lobes were never seen as a contraindication for treatment in the laser based procedures. Technically, laser treatment techniques such as side-firing transurethral coagulation, contact- and free-beam laser vaporization, interstitial laser coagulation, and the holmium laser-based resection and enucleation are fully suitable for treatment of median lobes. Surprisingly, no studies focussing specifically on laser treatment of median lobes have been published. PMID- 12084258 TI - Impact of interventional therapy for benign prostatic hyperplasia on quality of life and sexual function. AB - Considering the epidemiologic impact of benign prostatic hyperplasia (BPH), the use of minimally invasive surgery in the treatment of this condition is one of the main ambitions of contemporary urology. Compared with the prevalence of BPH and LUTS in elderly men, only a small percentage of patients requires surgery for indications such as renal insufficiency, chronic retention, and infection. The majority of patients seek treatment because of the bothersome nature of their symptoms and expect improvement in their quality of life. The goal of interventional therapy is to restore the comfort and well being of the patients, including preserving their sexuality. Therefore, there is increased focus on the quality of life and sexual function in the evaluation of the outcome of interventional therapy for BPH. PMID- 12084259 TI - Management of candiduria. AB - The distinction between Candida colonization of the urinary tract and infection is often blurred. Asymptomatic candiduria is particularly common in catheterized intensive care unit patients. To date, few studies have addressed the appropriate treatment regimens for candiduria. Fluconazole has become a mainstay of therapy; however, when to treat, whom to treat, and how long to treat are still largely unanswered questions. Asymptomatic nosocomial candiduria infrequently requires treatment intervention because morbidity is low and ascending infection and candidemia are rare complications. An understanding of the anatomic site of infection drives treatment decisions. More research is needed to define diagnostic criteria and therapeutic pathways. This review attempts to summarize the diagnosis and management of candiduria. PMID- 12084260 TI - Practice guidelines for the treatment of uncomplicated cystitis. AB - The Infectious Diseases Society of America has published guidelines for the treatment of uncomplicated cystitis. Recommendations are that for healthy, adult, nonpregnant women with bacterial cystitis, 3 days of trimethoprim/ sulfamethoxazole (TMP/SMZ) or trimethoprim alone is standard therapy in those regions where less than 10% to 20% of Escherichia coli that cause such infections is resistant to TMP/SMZ. In those regions where resistance is more than 10% to 20%, the committee recommended using an oral fluoroquinolone for 3 days, and that alternatives such as nitrofurantoin for 7 days or fosfomycin as single-dose therapy need more study. These recommendations were established in the late 1990s as resistance to TMP/SMZ among uropathogens was increasing in the United States, a phenomenon earlier observed in other parts of the world. Clinicians should be alert to patients infected with possibly resistant organisms, eg, patients who have recently been hospitalized or are receiving antibiotics. PMID- 12084262 TI - Renovascular hypertension in children. PMID- 12084261 TI - Urinary tract infections in the elderly. AB - The elderly population is now increasing in the world. A higher incidence of bacteriuria and urinary tract infection (UTI) is observed in elderly patients, in both long-term care facilities and at home. The management of elderly patients with UTI is increasing in clinical significance. Almost all of UTI in the elderly is complicated UTI. Control of the underlying diseases in the urinary tract is quite important in the management of UTIs in elderly patients. For pyelonephritis, switch therapy using aminoglycosides and fluoroquinolones, carbapenems, third-generation cephalosporines, or penicillins are selections of choice. The recommended duration of treatment for patients with pyelonephritis is 14 days. Seven to 10 days of treatment using fluoroquinolones or trimethoprim sulfamethoxazole is recommended for the treatment of elderly patients with symptomatic cystitis. Although asymptomatic bacteriuria is quite common in the elderly population, antibiotic treatment has no benefit for such patients. Intravaginal estrogen replacement is one of choice for the prevention of recurrent UTIs in postmenopausal women. PMID- 12084263 TI - Neoadjuvant androgen ablation before radical prostatectomy reduces positive margin rate, but has no effect on 36-month post-treatment biochemical failure. PMID- 12084264 TI - High grade prostatic intraepithelial neoplasia is a disease. AB - High grade prostatic intraepithelial neoplasia (PIN) is now widely accepted as the main premalignant lesion that has the potential to progress to prostate adenocarcinoma. High grade PIN is a standard diagnosis that must be included as part of the reported pathologic evaluation of prostate biopsies. Premalignant lesions that affect other organs have been identified and are treated when diagnosed such that the premalignant lesions itself are a disease (eg, carcinoma in situ of the bladder, colon polyps, and cervical dysplasia). Urologists should recognize that high grade PIN is a dangerous lesion and that it should be aggressively managed either by saturation biopsies of the prostate following the diagnosis of high grade PIN, or the more common recommendation--repeated prostate biopsies every 3 to 6 months for 2 years, then annually. Treatment of these precancerous lesions would appear to be of clinical benefit notwithstanding the potential for cancer prevention. These clinical benefits would reduce morbidity, enhance the quality of life, delay surgery or radiation, and increase the interval for surveillance requiring invasive procedures. PMID- 12084265 TI - Gene therapy for prostate cancer. AB - Basic research continues to unravel the molecular complexity of normal and abnormal biologic processes. The development of means to affect the expression level of genes that promote or contribute to cellular transformation, invasion, and metastasis has spawned the concept of gene therapy. This relatively new field seeks to reverse or suspend the pathologic progression of a variety of diseases including the malignant transformation of prostatic epithelial cells. Initial clinical trials for prostate cancer have thus far shown gene therapy to be relatively safe, although definitive evidence of durable therapeutic efficacy remains to be demonstrated. In this article, recent preclinical research, current therapeutic strategies, and recent results of gene therapy clinical trials for the treatment of prostate cancer are reviewed. PMID- 12084266 TI - Incontinence after radical prostatectomy: pathophysiology and management. AB - Despite improvements in surgical technique designed to preserve the functional integrity of the urethral sphincteric mechanism, incontinence after radical prostatectomy still occurs in many patients. Most patients have stress incontinence secondary to intrinsic sphincter deficiency, but many also have bladder dysfunction. The treatment of choice for post- prostatectomy stress incontinence is the artificial urinary sphincter. Other treatment options include collagen injection therapy and the male bulbourethral sling. PMID- 12084267 TI - Does the CaverMap device help preserve potency? AB - Erectile dysfunction remains a common complication following radical prostatectomy. The CaverMap Surgical Aid (UroMed, Boston, MA) was designed to aid the surgeon in identifying and preserving neurovascular bundles (NVBs). However, the size of the CaverMap nerve stimulator may make it difficult to trace the cavernous nerves before the prostate is removed, particularly in obese men or in patients who have a large prostate or a narrow pelvis. In a randomized, controlled study, the use of the CaverMap during radical prostatectomy resulted in improved nocturnal erections, but did not lead to improved overall sexual function. The CaverMap device, however, may be useful as a research tool in that it helps determine whether the NVBs have been successfully preserved after removing the prostate. However, preservation of the NVB does not guarantee recovery of potency, which may be prolonged despite successful stimulation of the cavernous nerves intraoperatively. This suggests that erectile dysfunction following radical prostatectomy is multifactorial. PMID- 12084268 TI - How to improve prostate biopsy detection of prostate cancer. AB - The combination of serum prostate-specific antigen (PSA) testing and transrectal ultrasonography is a highly effective strategy to diagnose prostate cancer at an early curable stage. Even though PSA is the most useful serum biomarker to aid in prostate cancer detection, it has limited specificity: as many as 75% of men who undergo prostate biopsy because of an elevated PSA do not have prostate cancer. Additionally, sextant prostate biopsies miss prostate cancer at least 20% of the time. To reduce the number of false-negative biopsies, many have advocated obtaining 12 or more cores in a single biopsy session. Studies have shown that this practice is safe and can enhance cancer detection modestly. Although it is unlikely that prostate cancer imaging will replace prostate biopsy in the near future, many exciting new imaging technologies should eventually improve targeting of prostate needle biopsy and reduce false-negative biopsies. Some of the most exciting areas include power Doppler sonography, microbubble intravenous ultrasound contrast agents, and magnetic resonance spectroscopy. These functional imaging modalities can assess tumor blood flow and metabolic activity at a cellular level and can detect malignant changes that may not be detected by standard anatomic imaging. PMID- 12084269 TI - Secondary hormonal manipulation of prostate cancer. AB - Prostate cancer is the second leading cause of cancer mortality among men in Western countries. The initial treatment of advanced prostate cancer is suppression of testicular androgen production by medical or surgical castration, but nearly all men with metastases develop disease progression. Patients with hormone-resistant prostate cancer (HRPC) have a median survival of approximately 18 months, and no therapy has yet demonstrated a definitive survival advantage. However, in the past several years, a number of promising new treatment strategies have emerged. One of the most important new treatment strategies involves secondary hormonal manipulation after the failure of primary androgen deprivation. This approach is predicated on the recognition that HRPC is a heterogeneous disease, and some patients may respond to alternative hormonal interventions despite the presence of castrate levels of testosterone. PMID- 12084270 TI - The role of complementary medicine in the treatment of prostate cancer. AB - Complementary and alternative medicine (CAM) has recently attracted national attention in the United States because of its widespread use and associated cost. CAM covers a broad spectrum of healing philosophies and approaches. Despite its widespread use, little is known about its safety, efficacy, cost effectiveness, and mechanism of action. The role of CAM in the management of prostate cancer is becoming more apparent with its rise in use among patients who are suffering from prostate cancer. This trend is patient-driven and reflects the change in values perceived by patients toward conventional medical treatment. In this review, several complementary therapies that can be used for prevention and treatment of prostate cancer are discussed. These therapies were selected because they are supported by scientific evidence. The popularity of CAM continues to grow and CAM is here to stay. Health care professionals can no longer afford to ignore or to treat CAM as an entity outside of conventional science. To acknowledge and to monitor its use among our patients may in the future help evaluate the safety and efficacy of CAM. Our current challenge is to move the field of CAM forward scientifically and systematically with wisdom and reasoning. PMID- 12084271 TI - Is it necessary to do staging pelvic lymph node dissection for T1c prostate cancer? AB - The necessity of pelvic lymph node dissection has been questioned in recent years as a result of improved pre-treatment staging based on clinical and pathologic factors. Accurate evaluation of nodal status allows rational selection of therapy and improved outcomes. Nevertheless, lymph node dissection may play a role even in patients with low stage disease (clinical T1c) despite an overall low risk for metastases. Herein we discuss recent advances in the evaluation of lymph nodes in stage T1c prostate cancer with respect to accurate prediction, radiologic imaging, molecular characterization, and operative considerations. PMID- 12084273 TI - Predicting continence following radical prostatectomy. AB - Stress urinary incontinence is a recognized complication following radical prostatectomy. Fortunately, in the hands of experienced surgeons, the overwhelming majority of men ultimately regain urinary continence following the procedure. Most men regain urinary continence 3 to 12 months after the prostatectomy. We have developed and validated a continence index that is administered at the time of catheter removal after radical prostatectomy. This index identifies those men who rapidly regain continence and men who will have permanent incontinence after prostatectomy. The study population was stratified into tertile groups based on the continence scores. At 3 months, 96%, 82%, and 68% of men in the highest, mid, and lowest tertile groups reported using no pads or one small pad. Based on these observations, we recommend initiating biofeedback immediately postoperatively in men with continence scores of 14 or less. At 1 year, 100%, 98%, and 87% of the men in the highest, mid, and lowest tertile group reported using no pads or only one small pad. Men who have continence scores greater than 14 can be assured that they will regain urinary continence within 1 year. To our knowledge, this index is the only validated instrument that predicts the return of urinary continence after radical prostatectomy. PMID- 12084272 TI - Immunotherapy of prostate cancer. AB - The realization that prostate cancer is an immunogenic tumor, in conjunction with the discovery of novel methods for priming the immune system to generate an antitumor response, has resulted in several new approaches for prostate cancer immunotherapy. Based on these various approaches, several human clinical trials have begun using immune-based therapies for prostate cancer. These approaches can be divided into cytokine-based therapies, tumor-associated antigen-based therapies, tumor vaccines, and dendritic cell-based therapies. This review summarizes the latest findings from each of these approaches and gives results from the few completed human clinical trials. PMID- 12084275 TI - Pediatric pyeloplasty. PMID- 12084276 TI - Vesicoureteral reflux and end-stage renal disease. PMID- 12084274 TI - Complexed prostate-specific antigen improvement in detecting prostate cancer. AB - This article presents the current reports of complexed prostate-specific antigen (PSA) aimed for the enhancement of prostate cancer detection. Further studies are needed to ascertain the variability of complexed PSA. Comparisons of percent free PSA, potential additive value of alpha(1)-antichymotripsin-bound PSA (PSA-ACT) and Bayer complexed PSA (cPSA) remains controversial in men with intermediate elevated total PSA concentration. Volume-referenced complexed PSA (PSA-ACT and cPSA) can enhance prostate cancer detection. Preliminary results show that PSA alpha(2)-macrobloblin (PSA-a(2)M) and PSA-alpha(1)-protease inhibitor (PSA-API) are promising assays for improving cancer detection. PMID- 12084277 TI - Management of the neonatal ureterocele. AB - Ureteroceles are potentially complex anomalies that have historically been managed with open reconstructive operations. In the last decade, endoscopic management has been proposed as an initial approach in patients with ureteroceles. With antenatal diagnosis, intervention can be performed on an outpatient basis prior to the development of any infections. Recent peer-reviewed studies help to define the appropriate technique, the patient population best managed with incision, and long-term outcomes. PMID- 12084278 TI - Management of neonatal ureteropelvic junction obstruction. AB - No topic in urology is more contentious than the management of neonatal ureteropelvic junction obstruction. Those favoring early diagnosis and correction of obstruction in early infancy cite excellent return of function and superior surgical results compared with delayed repair. Others believe that hydronephrosis improves or resolves with growth in many instances. In the most widely quoted series, almost all such patients improved. This article attempts to separate opinion from fact. The arguments for early surgery and for observation, even when obstruction has been diagnosed, are reviewed. Several experiences with long-term surveillance are summarized. PMID- 12084280 TI - Recent advances in hypospadias: current surgical technique and research in incidence and etiology. AB - Surgical techniques to repair hypospadias have undergone remarkable advances in recent years with the development and widespread use of dorsal urethral plate incision to facilitate urethroplasty. Clinical studies of the technique have shown excellent cosmesis and low complication rates for dorsal plate incision (DPI) in distal hypospadias repair, and have demonstrated the efficacy of expanding the use of the DPI in conjunction with other surgical techniques in more proximal repairs. There have been reports of increasing overall incidence of hypospadias in Scandinavian and US populations, but this might not be a uniform trend. There is, in fact, wide variation in the reported incidence of hypospadias among different countries. Many countries have not shown increased rates of hypospadias; furthermore, in countries noted previously to have an increased incidence of hypospadias, rates have leveled off since 1985. Investigations into the etiology of hypospadias have shown potential links to estrogen effects in utero from increased phytoestrogen intake in maternal vegetarian diets, and rare cases of genetic mutations in 5-alpha reductase type II. PMID- 12084279 TI - Management of vesicoureteral reflux in children. AB - Although much has been learned about the diagnosis and management of vesicoureteral reflux, several important areas of investigation remain. Because not all children with reflux are equally susceptible to renal scarring and the development of reflux nephropathy, controversy surrounds the need to evaluate all children with urinary tract infection or to continue prophylaxis in known refluxing children after a certain age. In addition to age, other factors such as sex, grade of reflux, and the presence of voiding dysfunction can all play a role. The grade of reflux as seen on the contrast voiding cystourethrogram is the best predictor of reflux resolution in large numbers of patients, but grade alone cannot predict spontaneous cessation in any one individual. Attempts at refining more quantitative imaging modalities have so far proved unsuccessful. Open ureteral reimplantation remains the standard for surgical care if surgery is necessary. Both cystoscopic and laparoscopic techniques, however, may ultimately prove to be reliable, minimally invasive approaches to definitive correction. Finally, there are data to support continued diagnosis and treatment of reflux in at-risk populations. The incidence of reflux-related morbidity in children has significantly diminished over the last three decades. A major challenge is to better identify at-risk subpopulations of children with reflux, so that not every child will require intensive, long-term medical treatment or surgery. PMID- 12084281 TI - Recent progress in the biology and treatment of Wilms' tumor. AB - The survival rate of patients with Wilms' tumor has rapidly improved in the last few decades. As we enter into the new millennium, overall survival for all Wilms' tumor patients exceeds 80%. Two large cooperative groups prospectively studied children with nephroblastoma: the National Wilms' Tumor Study Group, and the International Society of Paediatric Oncology. This review details the recent progress in understanding the biology of Wilms' tumor. Advances in clinical management of children with nephroblastoma are also outlined. PMID- 12084282 TI - Laparoscopy in pediatric urology. AB - Laparoscopy in pediatric urology is a rapidly evolving field that is becoming part of the operative repertoire of an increasing number of pediatric urologists. This article summarizes the latest ideas and issues in the expanding field of laparoscopy in pediatric urology. New methods of obtaining laparoscopic access and retraction are discussed. Laparoscopic experiences in urologic reconstruction, diagnosis and treatment of the nonpalpable testis, renal surgery, ureteral reimplantation, varicocelectomy, hydrocelectomy, and herniorrhaphy are reviewed. Articles assessing the safety of a pneumoperitoneum in patients with a ventriculoperitoneal shunt are also discussed. PMID- 12084283 TI - Posterior urethral valves: pre- and postnatal management. AB - Posterior urethral valves are a common problem encountered by pediatric urologists. The diagnosis is most frequently suggested by antenatal screening ultrasound. A variety of pre- and postnatal parameters have been identified to aid in predicting ultimate renal outcome. These prognostic tools are invaluable to the clinician for counseling parents and for choosing appropriate management. Several approaches to the treatment of patients with posterior urethral valves exist, and the ideal strategy is debatable. As technology evolves, more options for early intervention become available. Whether early detection and antenatal intervention improve patient outcome remains to be proven. PMID- 12084286 TI - Should laparoscopy be the standard approach used for radical nephrectomy? AB - Since its inception in June 1990, laparoscopic radical/total nephrectomy for renal tumor has been successfully applied worldwide to hundreds of patients. Recent 5-year follow-up data have shown this procedure to produce cancer control identical to that of open radical/total nephrectomy. Although in most centers the cost of the procedure remains higher than open surgery, the patient benefits of decreased pain, reduced hospitalization, less blood loss, and more rapid convalescence appear to be universal. At this time, we believe that laparoscopic radical/total nephrectomy for the treatment of renal tumors should become the new standard of care. PMID- 12084284 TI - The role of endourology in ureteropelvic junction obstruction. AB - Endopyelotomy has benefited from abundant confirmatory investigations, and significant progress in different technical modalities has occurred. Retrograde techniques, including the Acucise (Applied Medical, Laguna Hills, CA) cutting balloon and the ureteroscopic Holmium laser incision, are becoming preferred approaches while the other modalities retain their specific indications. Long term results and potential complications have been carefully studied and reported. Better identification of risk factors has prompted precise preoperative investigations and allowed for careful patient selection, leading to improved results. These results approach those of open pyeloplasty, but with minimal morbidity. PMID- 12084285 TI - A transperitoneal laparoscopic approach to endourology. AB - Although endoscopic methods have become the preferred means of management for many diseases facing the genitourinary surgeon, a laparoscopic approach might be considered comparable or advantageous in select circumstances. In the literature, laparoscopists reporting their work have favored the transperitoneal approach; however, there are clear advantages and disadvantages to both transperitoneal and retroperitoneal laparoscopy. Intracorporeal suturing remains the most time consuming aspect of reconstructive surgery, and research emphasis has been on suturing devices and novel anastomotic techniques. Laparoscopic pyeloplasty is efficacious and should be considered, particularly in the case of a capacious renal pelvis, crossing vessel, or failed previous endopyelotomy. Laparoscopic pyelolithotomy is uniquely suitable for patients with aberrant anatomy, such as a horseshoe kidney, and may be performed concurrently with pyeloplasty for ureteropelvic junction obstruction. The use of laparoscopic extravesical ureteral reimplantation awaits further development in both open and subtrigonal injection techniques. Its use in colposuspension is undetermined and requires further study as suturing technology improves. During laparoscopic exploration, it is possible to address intraoperative injuries to the ureter and bladder laparoscopically. In summary, laparoscopic surgery of the urinary tract is a "work in progress," but it offers promise for some of the most challenging of circumstances. As the technology advances and the clinical experience widens, the indications and contraindications for these techniques will be better established. PMID- 12084288 TI - The treatment of erectile dysfunction in the elderly. PMID- 12084287 TI - Should laparoscopy be the standard approach used for pelvic lymph node dissection? AB - Involvement of the pelvic lymph nodes in patients with prostate cancer worsens the overall prognosis of this common disease entity. Prior radiographic staging techniques, including fine-needle aspiration, are limited by a poor sensitivity and are not reliable. The gold standard for the evaluation of pelvic lymph nodes in men with prostate cancer involves performing a lymphadenectomy. Historically, this procedure was performed using an open surgical technique. Unfortunately, this invasive procedure is associated with significant morbidity. In response, modern surgical technology has provided newer, less invasive techniques, including laparoscopic pelvic lymphadenectomy (LPLND). Improved detection of localized prostate cancer through the institution of screening protocols and early detection programs has decreased the number of patients presenting with lymph node involvement. Various clinical indicators, including prostate-specific antigen, grade, and stage, have been used to improve the selection of "high-risk" patients that are appropriate candidates for pelvic lymph node dissection. The technique of LPLND is a valid option in the armamentarium for staging of prostate cancer. The laparoscopic approach provides the same staging accuracy as the open surgical technique and is superior with respect to morbidity. LPLND is limited to patients who present with a high risk of advanced prostate cancer. In addition, the urologist must accept the additional training, financial expense, and "learning curve" associated with this technique. PMID- 12084289 TI - Antenatal diagnosis and treatment of congenital adrenal hyperplasia. AB - Congenital adrenal hyperplasia (CAH) is a family of monogenic autosomal recessive disorders of steroidogenesis in which enzymatic defects result in impaired synthesis of cortisol by the adrenal cortex. The adrenal 21-hydroxylase (21-OH) enzyme is one of five enzymes necessary for the synthesis of cortisol from cholesterol, and its deficiency is the most common enzymatic defect causing CAH. 21-OH deficiency (21-OHD) occurs in a classical form that can cause genital ambiguity at birth in genetic females. Newborn males have normal genitalia. Prenatal treatment of 21-hydroxylase deficiency with dexamethasone has been used for approximately 15 years. An algorithm was developed for prenatal diagnosis and treatment. PMID- 12084291 TI - Early identification of patients with von Hippel-Lindau disease at risk for pheochromocytoma. AB - von Hippel-Lindau disease (VHL) is an autosomal dominant familial syndrome that predisposes to the formation of tumors in multiple organ systems, including adrenal and extra-adrenal pheochromocytomas. However, fewer than 30% of VHL families develop pheochromocytomas. In recent years, this clinical heterogeneity has been correlated with missense mutations. The VHL patient requires vigilant, lifelong biochemical and radiographic screening for pheochromocytoma. Half of VHL pheochromocytomas present bilaterally, and there is a high incidence of recurrence after surgery. Because of the morbidity of bilateral total adrenalectomy with subsequent steroid replacement therapy, the recent therapeutic trend has been toward observation and minimally invasive adrenal-sparing procedures. PMID- 12084290 TI - Partial adrenalectomy in patients with multiple adrenal tumors. AB - Most adrenal tumors are found incidentally and appear as small solitary nodules on abdominal imaging. Occasionally, work-up demonstrates multifocal or bilateral adrenal tumors. Certain patients are predisposed to multiple lesions, such as those with hereditary forms of pheochromocytoma as seen in von Hippel-Lindau disease, multiple endocrine neoplasia type II, and von Recklinghausen's disease. Partial rather than total adrenalectomy should be considered for these patients in an attempt to preserve endogenous adrenocortical function. Partial adrenalectomy has also been used to resect other types of adrenal tumors, especially in patients with a solitary adrenal gland. A discussion of the indications for partial adrenalectomy and of the surgical technique follows. PMID- 12084292 TI - Phase I clinical trial of interleukin 2 (IL-2) gene therapy for prostate cancer. PMID- 12084294 TI - Laparoscopic nephrectomy for renal cell carcinoma. AB - Since the first reported case of laparoscopic nephrectomy by Clayman et al. in 1991, laparoscopy is gaining acceptance as a viable alternative to open surgery for renal cell carcinoma. The benefits of laparoscopy include improved quality of life and lower incidence of perioperative morbidity. The perceived risks of laparoscopic nephrectomy for renal cell carcinoma include port-site metastasis, increased operative time, and the concern for inadequate surgical resection. The preliminary data concerning laparoscopy in renal cell carcinoma, however, indicate that rates of tumor recurrence are equivalent to open surgery while resulting in better cosmesis, decreased level of perioperative analgesic use, and decreased length of time to full convalescence. PMID- 12084293 TI - Nephron-sparing surgery. AB - The curative management of renal cell carcinoma remains surgical. Recent advances in imaging and increased use of cross-sectional imaging modalities have led to an increased detection of incidental renal cell carcinomas. There is little debate regarding the role of nephron-sparing surgery (NSS) when absolutely indicated. Radical nephrectomy is still considered by many as the treatment of choice for localized renal cell carcinoma in the setting of a normal contralateral kidney. However, there is growing evidence that in the correct patient, the use of NSS in this setting is justified. Therefore, the indications of NSS have evolved in the past decade. Recent data indicate that radical nephrectomy and NSS provide effective and equivalent curative treatment for most renal cell carcinomas, especially those 4 cm or smaller. These data, along with new, refined surgical techniques, new studies regarding the biology of renal cell carcinoma and true incidence of occult multifocality, and earlier diagnosis make NSS an attractive consideration for the practicing urologist. PMID- 12084295 TI - Dendritic cell-based immunotherapy of renal cell carcinoma. AB - Although mostly resistant to cytotoxic therapy, renal cell carcinoma has been a testing ground for immunotherapy. The approval of interleukin-2 for the treatment of renal cell carcinoma was a landmark "proof of principle" which showed that agents working solely via the immune system can cause durable cancer remission. Dendritic cells are central to immune-mediated surveillance and destruction of abnormal cells. They possess all the components required to educate immune effector cells that can then mediate tumor destruction. In vitro strategies to expand and load dendritic cells with antigens have now led to human vaccine trials in renal cell carcinoma and other malignancies. PMID- 12084296 TI - Molecular-based therapies for renal cell carcinoma. AB - The incidence of renal cell carcinoma (RCC) is rising steadily, but the ability to cure patients with metastatic RCC unfortunately remains limited. Emerging interest in gene therapy performance and safety is expressed by patients, medical institutes, and other agencies. It has become evident that better understanding of the genetic impairments and immune pathophysiology in RCC is essential for future improvement in patient care. Clinical trials now underway that are focusing on genetic and immune impairments will hopefully lead to future breakthroughs in RCC therapy. This paper reviews available gene therapies and other related therapeutic approaches for RCC and lists some of the current clinical trials focused on molecular-based therapies. PMID- 12084297 TI - Immunotherapy for bladder cancer. AB - The primary role of immunotherapy for bladder cancer is to treat superficial transitional cell carcinomas (ie, carcinoma in situ, Ta, and T1). Immunotherapy in the form of bacille Calmette-Guerin (BCG), interferon, bropirimine, keyhole limpet hemocyanin, and gene therapy is intended to treat existing or residual tumor, to prevent recurrence of tumor, to prevent progression of disease, and to prolong survival of patients. Presently, BCG is commonly used and is the most effective immunotherapeutic agent against superficial transitional cell carcinoma. Data support that BCG has a positive impact on tumor recurrence, disease progression, and survival. Proper attention to maintenance schedules, route of administration, dosing, strains, and viability is essential to obtain the maximum benefits of BCG immunotherapy. This review highlights and summarizes the recent advances concerning immunotherapy, with special emphasis on BCG therapy for transitional cell carcinoma. PMID- 12084298 TI - Prostate cancer genomics. AB - The molecular processes contributing to cancer of the human prostate gland are under intensive investigation. Methods used for discovering genetic alterations involved in prostate neoplasia include family studies designed to map hereditary disease loci, chromosomal studies to identify aberrations that may locate oncogenes or tumor suppressor genes, and comprehensive gene expression studies. These studies determine how various molecular signaling pathways influence or reflect the process of carcinogenesis. However, a comprehensive overview of the cell is necessary to understand all of the dynamic interactions between genes, their protein products, and the network of cellular processes resulting in tumorigenesis. Unraveling the complexity of these systems in a timely manner involves the integration of computers, miniaturization, and automation into molecular biology. New biotechnologies such as the development of automated DNA sequencing and complementary DNA microarrays allow for a systematic, "discovery driven" approach. These and other technologies afford a comprehensive view of biology and pathology that have the potential to fully characterize the processes involved in neoplasia and therefore provide potential targets for the therapy of prostate and other cancers. PMID- 12084299 TI - Fibrin glue applications in urology. AB - Fibrin glue is used as a hemostatic and adhesive agent. Until recently, safety issues had delayed approval of the commercial preparation in the United States. Fibrin glue has found several urologic applications, especially as an adhesive agent. It has excellent potential in laparoscopic surgery, where conventional tissue approximation techniques are cumbersome and time-consuming. Fibrin glue has also been used as a delivery vehicle for autologous cell transplantation and may play a major role in tissue engineering techniques in the future. PMID- 12084300 TI - Tissue engineering in urology. AB - Congenital abnormalities, cancer, trauma, infection, inflammation, iatrogenic injuries, and other conditions may lead to genitourinary organ damage or loss, requiring eventual reconstruction. Tissue engineering follows the principles of cell transplantation, materials science, and engineering toward the development of biological substitutes that would restore and maintain normal function. Tissue engineering may involve matrices alone, wherein the body's natural ability to regenerate is used to orient or direct new tissue growth, or the use of matrices with cells. Both synthetic (polyglycolic acid polymer scaffolds alone and with co polymers of poly-1-lactic acid and poly-DL-lactide-coglycolide) and natural biodegradable materials (processed collagen derived from allogeneic donor bladder submucosa and intestinal submucosa) have been used, either alone or as cell delivery vehicles. Tissue engineering has been applied experimentally for the reconstitution of several urologic tissues and organs, including bladder, ureter, urethra, kidney, testis, and genitalia. Fetal applications have also been explored. Recently, several tissue engineering technologies have been used clinically, including the use of cells as bulking agents for the treatment of vesicoureteral reflux and incontinence, urethral replacement, and bladder reconstruction. Recent progress suggests that engineered urologic tissues may have clinical applicability in the future. PMID- 12084302 TI - Germ cell transplantation: a potential treatment of severe testicular failure. AB - Although the process of spermatogenesis is relatively efficient and resistant to damage, male infertility can result from exposure to toxic agents such as chemotherapeutic regimes, radiation, or occupational exposures to chemicals. Other types of infertility may result from migratory defects or poor survival of primordial germ cells during development, abnormal repopulation of the tubules by spermatogonia during development, or low cellularity of the testis (hypospermatogenesis). Presently, there are no effective therapies available to treat these patients. Recent studies in animal models have demonstrated that isolated testicular germ cells collected from testes may be transplanted into sterile recipient mice to regenerate spermatogenesis. This technology will have widespread applications in efforts to manipulate the genome and produce transgenic offspring, to improve agricultural species, to enhance sperm production in endangered species, to improve our understanding of the control mechanisms regulating spermatogenesis, and to treat male infertility. PMID- 12084301 TI - The possible effects of environmental estrogen disrupters on reproductive health. AB - The term environmental estrogen refers to chemical substances that exhibit some degree of estrogen-like activity. The primary emphasis for potential adverse effects resulting from exposure to environmental estrogens is on in utero exposure because such exposures can occur during critical periods of organogenesis. Assessment of biological plausibility can be based, in part, on the extensive data on the effects of diethylstilbestrol (DES). The available evidence is too limited to judge with any confidence whether sperm counts have declined during the past 50 years. Based on both animal and human data with DES, it is biologically plausible that in utero exposures to exogenous estrogenic compounds are capable of reducing sperm production in adult men. However, the apparent existence of a maternal dose threshold for DES-induced effects on sperm count undermines the likelihood that environmental estrogens, which are substantially less potent, are capable of causing similar effects. PMID- 12084303 TI - Diagnostic and therapeutic testis biopsy. AB - Testicular biopsy has been traditionally used as a diagnostic tool in the management of infertility. With the advent of assisted reproductive technology, testicular biopsy is now used therapeutically to retrieve sperm for intracytoplasmic sperm injection (ICSI). The feasibility of testicular sperm extraction for ICSI is reviewed in this article. Although less invasive techniques such as testicular fine needle aspiration and percutaneous needle biopsy are efficacious, particularly in cases of obstructive azoospermia, studies in the current literature support that open testicular biopsy is a more reliable method to obtain testicular specimen. Different measures to reduce the complication rate of open testicular biopsy, including the application of microsurgical techniques, are addressed here. Other areas of recent development related to testicular biopsy that are reviewed include processing of testicular tissue, cryopreservation of retrieved spermatozoa, in vitro maturation of spermatozoa, and microinjection of immature spermatogenic precursor cells. PMID- 12084305 TI - Current status of standardized questionnaires in the measurement of erectile dysfunction. AB - The evaluation and management of erectile dysfunction has evolved dramatically over the past decade. Clinicians now benefit from advances in the understanding of the physiologic pathways involved in the function of erections and from the availability of innovative and effective therapies. The clinical history remains the essential starting point in the evaluation of patients with erectile dysfunction. The standardized questionnaire has been used as a valuable complement to the clinical history and in the assessment of treatment efficacy. PMID- 12084304 TI - Genetic causes of male infertility: current concepts. AB - Infertile men with severe spermatogenic defects and low or no sperm counts have a significantly higher rate of genetic abnormalities than fertile men. The fact that intracytoplasmic sperm injection can potentially bypass natural selection barriers to genetic disease transmission has brought a sobering but important impetus to recent research in the area of genetic infertility. Recent studies have focused on examining the prevalence of certain genetic defects in infertile men, analyzing the molecular basis of infertility in genetic disorders, and detecting new causes of genetic infertility. Several novel research findings deserve mention for their potential impact on genetic infertility. It has been demonstrated that elongated and round spermatids can be successfully injected into human oocytes and viable births obtained. Likewise, significant advances have been made in the arena of interspecies germ cell transplantation. Of some concern is the finding of a relationship between faulty DNA repair and infertility in men with severe testis failure. This review summarizes the recent genetic advances in these areas of male genetic infertility. PMID- 12084306 TI - Current status of penile prosthesis implantation. AB - Today most men with erectile dysfunction (ED) can be effectively treated with less invasive means than penile prosthesis implantation. Nevertheless, there remain men who do not respond to simple treatments such as systemic medication and who either do not respond to or reject intermediate treatment such as penile injection therapy. For these men, penile prosthesis implantation remains a reasonable option. This review discusses types of penile prostheses, how to choose among prosthesis types, key features of implant techniques, infection management, penile reconstruction related to prosthesis implantation, and patient and partner satisfaction. PMID- 12084307 TI - Peyronie's disease: advances in basic science and pathophysiology. AB - Peyronie's disease is an idiopathic, localized connective tissue disorder of the penis that involves the tunica albuginea of the corpus cavernosum and the adjacent areolar space. The tunica albuginea plays an important role in the mechanism of erection. Peyronie's disease is characterized by local changes in the collagen and elastic fiber composition of the tunica albuginea. The formation of fibrotic plaques alters penile anatomy and can cause different degrees of bending, narrowing, or shortening of the penis. Moreover, a significant number of men with Peyronie's disease develop erectile dysfunction. Penile blood flow studies in many patients with Peyronie's disease suggest a strong association with veno-occlusive dysfunction. Although long recognized as an important clinical entity of the male genitalia, the etiology of this disease has remained poorly understood. The following review focuses on recent research on the pathophysiology of Peyronie's disease. PMID- 12084308 TI - Rapid ejaculation. AB - Rapid ejaculation, or premature ejaculatory dysfunction, is the most frequently encountered sexual complaint of men and couples. It is most common in adolescents, young adults, and other sexually naive males. Increased risk is associated with lack of sexual experience, lack of knowledge regarding normal male and female sexual responses, and with those individuals who highly associate psychological factors (such as fear, guilt, and anxiety) with sexual activity. This article provides a basic review of the pathophysiology of ejaculatory dysfunction and discusses the current treatment options for rapid ejaculation. PMID- 12084309 TI - Oral and injectable medications for the treatment of erectile dysfunction. AB - The treatment of erectile dysfunction has changed dramatically over the past two decades. The introduction of the oral agent sildenafil 2 years ago has revolutionized the treatment of men with compromised erections and has met with expected success and low morbidity. Sildenafil is effective in most men with erectile dysfunction in the general population and in select populations, such as men with spinal cord injury, diabetes mellitus, and patients who have had nerve sparing radical prostatectomy. It is safe in the general population as well as in many men with cardiac disease. Other newer medications are in trial and may soon be available to supplement treatment with sildenafil. Oral phentolamine, apomorphine, newer phosphodiesterase type-5 inhibitors, and topical agents are currently in phase 3 trials. These agents, in addition to newer intraurethral and indictable agents, may assist men with erectile dysfunction and rescue those in whom sildenafil is ineffective or in whom untoward side effects of sildenafil reduce its effectiveness. The 21st century will witness many additional agents designed for specific patients with specific conditions causing erectile dysfunction. We can expect these oral agents, assisted by topical and injectable agents, to successfully restore erectile function in the majority of men suffering from erectile dysfunction. PMID- 12084310 TI - Androgen deficiency in the aging male: when to evaluate and when to treat. AB - Aging can be associated with an abnormal decline in testosterone production, mainly due to pituitary failure, and is reflected by low serum testosterone concentrations. Manifestations of testosterone deficiency can be subtle, can be misinterpreted as consequences of normal aging, and include lack of libido and energy, mood changes, impaired strength, and development of osteoporosis. The most important factor in the evaluation and treatment of this age-related, usually unrecognized abnormality is the public awareness of its frequent occurrence that will stimulate the use of appropriate screening tests, beginning with a morning testosterone level. Replacement therapy by parenteral or transdermal testosterone administration can be highly beneficial in alleviating the manifestations of hypogonadism and improving quality of life. Careful screening and meticulous follow-up are mandatory. The long-term effects of testosterone replacement on the cardiovascular system, the prostate, and on longevity are unknown. PMID- 12084311 TI - Which patients with transitional cell carcinoma of the bladder or prostatic urethra are candidates for an orthotopic neobladder? AB - Bladder replacement has become a standard method of urinary diversion in adults undergoing cystectomy. When faced with a treatment decision, the patient and urologist are most comfortable if information is available on short- and long term possible outcomes of the proposed intervention and on the magnitude of the expected effect. This article presents and discusses absolute and relative contraindications for orthotopic neobladders that may help optimize the clinical results with these procedures. As our own series has progressed, so has our philosophy regarding patients who are candidates for an orthotopic reconstruction. All men who require radical cystectomy as treatment for bladder cancer are initially thought to be appropriate candidates for orthotopic lower urinary tract reconstruction. Given the criteria presented in this article approximately, 80% of men are considered acceptable candidates for a neobladder. By these criteria, 65% of women are adequate candidates for an orthotopic bladder. PMID- 12084312 TI - Optimal management of the 3- to 6-centimeter anterior urethral stricture. AB - The optimal management of anterior urethral stricture that does not respond to an endoscopic urethrotomy or is found to be unsuitable for excision and anastomotic repair remains controversial. Genital skin island onlay flaps or buccal mucosal grafts are presently the most dependable single stage procedures used for strictures more than 3 cm in length. Nonhirsute penile island fasciocutaneous flaps constitute the most durable substitution technique for pendulous stricture disease, with long-term studies reporting 90% to 96% success. The complex proximal bulb and bulbomenbraneous stricture with a compromised proximal fibrous avascular bed is ideally managed with either a penile or scrotal island flap or some combination of partial urethral excision with a dorsally placed genital skin island. The buccal mucosal onlay graft is a promising addition to this reconstructive paradigm, and early outcomes have been favorable. The graft is presently used for bulbar strictures, avoiding the transsphincteric on pendulous location, or a compromised recipient bed. The present standard of care for proximal bulb strictures is wide bulbospongiosal mobilization, partial urethral excision, a floor strip anastomosis, and placement of an augmenting flap on the graft in a dorsal location. PMID- 12084313 TI - Current investigations and treatment of interstitial cystitis. AB - Interstitial cystitis (IC) is a chronic condition characterized by a constellation of symptoms such as urinary frequency, nocturia, urinary urgency, suprapubic pressure, and bladder and pelvic pain. Since its original description, the etiology of the disorder has remained unknown despite intense investigations. The International Cystitis Association (ICA) and the National Institutes of Arthritis, Diabetes, Digestive and Kidney Diseases (NIDDK) have been instrumental in supporting the United States Interstitial Database (ICDB) and foster research to study the disorder. The NIDDK developed criteria to ensure that all groups of patients treated would be relatively comparable. However, many patients who would be clinically considered to have IC do not fulfill all the NIDDK criteria. Many clinical criteria for the diagnosis of IC, such as the presence of glomerulations and the intravesical potassium chloride test, are being challenged. The epidemiology of the disorder is not well established, but there are an estimated 700,000 cases of IC in the United States. Numerous pathophysiologic mechanisms have been proposed, but none have been proven. There is no representative animal model of IC. Both the oral and intravesical treatments of IC are noncurative, and few are based on a plausible mechanism or scientific evidence. Surgical treatment should be considered with extreme caution; it is the last therapeutic option because failure rate can be substantial. PMID- 12084314 TI - Neurostimulation for lower urinary tract voiding problems. AB - In 1981, Tanagho and Schmidt introduced chronic electrical stimulation of the sacral spinal nerves using a permanently implanted sacral foramen electrode and a battery-powered pulse generator for treatment of different kinds of lower urinary tract dysfunction, refractory to conservative treatment. At our department, chronic unilateral electrical stimulation of the S3 sacral spinal nerve has been used for treatment of vesicourethral dysfunction in 55 patients with a mean postoperative follow-up of 44.3 months. Lasting symptomatic improvement of more than 50% was achieved in 16 of 21 patients with motor urge incontinence (76.2%) and in 22 of the 28 patients with urinary retention (78.6%). In our opinion, chronic sacral neuromodulation offers a sustained therapeutic effect to treatment responders that is not achieved by temporary neuromodulation techniques. Chronic neuromodulation should be considered predominantly in patients with urinary retention and in patients with motor urge incontinence who refuse temporary techniques or who require too much effort to achieve a sustained clinical effect. PMID- 12084315 TI - The role of a second transurethral resection for high-grade bladder cancer. AB - High-grade, non-muscle-invasive bladder cancer poses many difficult diagnostic and therapeutic challenges. The superficial, yet high-risk, nature of disease progression exemplifies the unique behavior of this form of bladder cancer. An adjunct to managing this difficult problem is a second staging transurethral resection of the bladder tumor (TURBT). This article reviews the pros and cons of a second staging TURBT and how it may impact the management of high-grade, low stage superficial bladder cancer. PMID- 12084316 TI - Use of collagen for the treatment of stress urinary incontinence: an update. AB - This article provides a comprehensive and updated overview of the current role of collagen injections in the management of genuine stress incontinence. The clinical indications for collagen injection are described, and relevant technical advances are discussed. Finally, we review the latest outcome data on the use of collagen injection, as well as the factors that affect outcome. PMID- 12084317 TI - Current concepts and controversies in urodynamics. AB - Urodynamics is the dynamic study of the transport, storage, and evacuation of urine by the urinary tract. It is comprised of several tests that, when used individually or collectively, can give information about lower urinary tract function. The components of the urodynamic study are uroflowmetry, cystometry, pressure-flow studies, electromyography, urethral pressure profilometry, leak point pressure measurement, videourodynamics, and ambulatory urodynamics. Familiarity with the recent advances and controversies of each component is essential when using urodynamics to diagnose and treat lower urinary tract dysfunction. PMID- 12084320 TI - Benign prostatic hyperplasia: many new options, same old approach. PMID- 12084318 TI - Lower urinary tract physiology and pharmacology. AB - This review focuses on what we consider to be the most important findings of the last year relating to the smooth muscle of the lower urogenital system and the different levels of regulation that control its contraction and relaxation. One level is through modulation of the smooth muscle itself or its environment. Recent findings examining myosin isoform composition and collagen content as well as mechanisms that appear to be involved in inducing hyperplasia/hypertrophy of smooth muscle are described. Another method of regulation is via calcium dependent phosphorylation of the regulatory light chain of myosin, which increases its activity. Interesting results indicating an uncoupling of force from calcium in the bladder are discussed. A third level of regulation is pharmacologic. Thus, the most recent findings related to receptor subtypes, including muscarinic, endothelin, alpha-adrenergic and nicotinic receptors, are presented. In addition, the effects of diabetes, incontinence, and partial bladder outlet obstruction on these modes of contractile regulation are also discussed. PMID- 12084319 TI - Treatment of urinary incontinence. AB - The treatment of urinary incontinence is a dynamic and evolving field. New therapies, techniques, and procedures, as well as some subtle refinements in treatments currently used, offer hope for the millions of people suffering from this condition. Recent attention has been directed toward reevaluating the efficacy and durability of some standard treatments for stress urinary incontinence, including pelvic floor exercise, bladder neck suspensions, and pubovaginal slings. Occlusive or supportive devices have evolved into a suitable alternative in some patients. Extracorporeal magnetic therapy and alternative periurethral injectable agents may offer additional treatment strategies for the relief of symptomatic stress urinary incontinence. New drugs for the treatment of overactive bladder and urge urinary incontinence have been introduced recently and, in combination with neuromodulation, offer the first new treatments for this condition in over 25 years. Yet, as rapidly as new therapies become available for the treatment of urinary incontinence, problems have become evident with some that were introduced just a short time ago. Thus, cautious skepticism regarding these "new and improved" treatments should be maintained until long-term data become available with respect to safety, efficacy, and durability. PMID- 12084321 TI - Induction of apoptosis in the prostate by alpha1-adrenoceptor antagonists: a novel effect of "old" drugs. AB - Advances in our understanding of the integrated functions governing prostate cell proliferation and cell death imply that effective therapies for prostate neoplasia should not only be molecularly targeted, but should be customized to take into account the delicate balance of opposing growth influences. Evidence from studies on the dynamics of prostate growth in benign prostatic hyperplasia (BPH) and prostate cancer established that disruption of the molecular mechanisms that regulate apoptosis and cell proliferation among the stroma and epithelial cell populations may underlie the neoplastic development that characterizes the aging gland. Our own efforts have been focused on investigating whether a1 adrenoceptor antagonists clinically used for the relief of the obstructive symptoms associated with BPH affect prostate pathophysiology via mechanisms other than smooth muscle contraction. Such efforts led to the identification of a novel effect of two alpha1-adrenoceptor antagonists, doxazosin and terazosin. More recent in vitro experiments examined the potential anti-tumor action of three clinically used alpha1-adrenoceptor antagonists--doxazosin, terazosin and tamsulosin--against prostate cancer cell growth. These findings demonstrate the ability of doxazosin and terazosin, but not tamsulosin, to suppress prostate growth by inducing apoptosis among the epithelial cells in the benign and malignant prostate. Thus, evidence indicates that rather than just causing pure relaxation of the smooth muscle, certain alpha1-blockers can also affect the dynamics of prostate growth by changing the balance between prostate cell proliferation and apoptosis at the expense of the proliferative process. PMID- 12084323 TI - Phytotherapeutic agents in the treatment of benign prostatic hyperplasia. AB - The rationale and efficacy of phytotherapeutic agents in the treatment of lower urinary tract symptoms secondary to benign prostatic hyperplasia (BPH) are continuously debated. While plant extracts are prescribed and reimbursable treatment options in Europe, they are officially classified merely as dietary supplements in the United States. The most commonly used preparations originate from the species Serenoa repens, Pygeum africanum, hypoxis rooperi, pinus, picea, urtica dioica, and secale cereale. Combination extracts derived from two or more plants are also used. Various components have been suggested to be active, and different mechanisms of action are being supposed. Open trials and some short term randomized studies, suggesting safety and efficacy, have been reported. However, if stringent criteria of evidence-based medicine are applied, the data are inconclusive. Therefore, the 4th International Consultation on BPH and the recent German guidelines have not (yet) recommended phytotherapy for the management of symptomatic BPH. PMID- 12084324 TI - Transurethral microwave thermotherapy in benign prostatic hyperplasia. AB - This article reviews the available literature and data on high-energy transurethral microwave therapy (TUMT) in the treatment of benign prostatic hyperplasia (BPH) causing lower urinary tract symptoms (LUTS). TUMT is a safe, durable, (1-hour) procedure, without the need for anesthesia. Emphasis is made on the effect and mechanism of TUMT, the different devices available including different energy protocols, and accompanying clinical results. PMID- 12084322 TI - The clinical role of alpha-blockers in the treatment of benign prostatic hyperplasia. AB - Benign prostatic hyperplasia (BPH) is a highly prevalent condition that represents a significant health problem, which is likely to worsen as the population continues to age. alpha-1 adrenoceptor antagonists have been used in the treatment of this condition for well over 20 years and much is now known about their effects on the urinary tract. As these drugs have improved, their use as a first line treatment option in patients with lower urinary tract symptoms (LUTS), secondary to BPH, has continued to increase. This review details some of the background and rationale for the use of these drugs and, using the available information, compares their efficacy and tolerability profiles. PMID- 12084325 TI - Transurethral vaporization of the prostate: current techniques. AB - Electrovaporization has quickly become a popular procedure for treating benign prostatic hyperplasia. By using standard transurethral electrosurgical technology, electrovaporization offers standard transurethral loop-like resection and transurethral prostatectomy (TURP)-like efficacy with less morbidity than that associated with TURP. Through a combination of two electrosurgical effects (vaporization and desiccation), electrovaporization ablates significant volumes of prostatic tissue with each passage of the electrode. Newly designed thicker loop electrodes have led to a modification of the original electrovaporization technique. "Vaporizing-resection" adds the advantages of transurethral resection to those of electrovaporization with the use of one electrode. Very large glands can be treated efficiently, and prostatic tissue specimens are produced. Moreover, additional vaporization of tissue occurs with minimized bleeding. This report reviews basic electrosurgical principles, electrode design, and factors that affect vaporizing efficiency. Current long-term data on electrovaporization compared with TURP are summarized, and pilot studies evaluating vaporizing- resection have shown promising results. A new technology that has entered the market--bipolar electrovaporization--is also discussed. Finally, combination therapy and Columbia's 4-year experience with vaporization techniques are discussed. PMID- 12084326 TI - The use of lasers in benign prostatic enlargement. AB - This article reviews the laser techniques available for the treatment of lower urinary tract symptoms. Early data on short-term mortality, morbidity, complication rates, and outcome are available, but information on cost effectiveness, long-term outcome, and patient preference are as yet unavailable. In the 8 years since laser methods became popular it has become clear that substantial and variably durable flow and symptomatic responses are achieved that do not quite match transurethral resection of the prostate (TURP). Less immediate physiologic stress is balanced by a delay before full response. Same-day surgery may be possible, but an extended period of catheterization may be necessary. Laser methods remain an option for treatment and are particularly applicable to the less fit man. Whether men will choose a less effective but less invasive treatment option for symptom relief is not yet obvious. Limited early data suggest the holmium laser technique most closely approaches the gold standard. PMID- 12084327 TI - Epidemiology of prostatitis. AB - Prostatitis is the third most important condition of the prostate. Problems in the diagnosis and classification of this condition, however, have hindered epidemiologic research. Consequently, our understanding of the incidence of prostatitis and factors that increase its probability is limited. Recent studies suggest that the prevalence of prostatitis is 5% to 9% among unselected men in the community. Based on estimates from national data, approximately 2 million men in the United States seek treatment for prostatitis each year. A number of recent studies suggest that genetic, behavioral, and environmental factors; age; inflammatory mediators; and oxidative stress may be risk factors for prostatitis. However, findings from most of these studies may at best be considered preliminary because of problems with small sample sizes or lack of generalizability. Nonetheless, these studies suggest hypotheses that should be tested in well-designed population-based studies. PMID- 12084328 TI - An evidence-based approach to the treatment of prostatitis: is it possible? AB - The classification of prostatitis has been re-evaluated, and the National Institutes of Health (NIH) has defined the separate groups more thoroughly. Furthermore, the development of the NIH chronic prostatitis symptom index (CPSI) has allowed the symptoms to be measured and the effect of interventions calculated. A search of the literature finds that the quality of treatment trials in prostatitis is poor and the level of evidence for many of the existing strategies is lacking; there is a total absence of any meta-analyses of randomized controlled trials (level 1a). The bulk of literature consists of small descriptive studies (level III) or opinions from respected authorities (level IV). The advent of the NIH-CPSI has provided the opportunity for well-designed trials to be performed where the symptom outcomes can be quantified. This opportunity will be of greatest value in type III prostatitis where symptom amelioration is the main target of therapy and a number of treatment options abound. PMID- 12084329 TI - Alpha-blockers: an effective treatment for prostatitis? AB - Chronic prostatitis is a multifaceted problem affecting men that is both poorly understood and poorly treated. It is inevitable, therefore, that no single treatment will benefit all patients. However, data suggest that some patients' problems can be treated effectively with alpha-blockers. The prostate is rich in alpha-adrenergic receptors, and some urodynamic studies have shown a failure of the bladder neck to relax during voiding. Other studies have demonstrated turbulent flow in the prostate leading to prostatic duct reflux. Because urine is acidic, reflux into the prostatic ducts will cause inflammation and pain. Thus alpha-blockers are an effective treatment for men with prostatitis caused by dysfunctional voiding. PMID- 12084330 TI - Pelvic floor physical therapy for patients with prostatitis. AB - The most common form of prostatitis is National Institutes of Health category III, also known as chronic abacterial prostatitis/chronic pelvic pain syndrome. The search for effective and durable therapy for this condition remains frustrating for both patients and physicians. A new approach incorporating a neurobahavioral and musculoskeletal perspective is emerging as a means of diagnosing and treating affected patients. Adopting methods from gynecology, colorectal surgery, and physical therapy has had promising effects in men diagnosed with chronic prostatitis. PMID- 12084331 TI - Microbiology of the prostate. AB - Prostatitis is a common genitourinary disease in men. The National Institutes of Health has defined four types of prostatitis. The bacteriology of acute and chronic bacterial prostatitis is well understood. Causative organisms can be isolated by four-glass urine cultures. Chronic pelvic pain syndrome (CPPS; chronic abacterial prostatitis) is an enigma; whether there is a microbiologic cause for this condition is unknown. Researchers have postulated that CPPS may be an infectious disease of the prostate. This article reviews current literature regarding the microbiology of the prostate in CPPS. PMID- 12084332 TI - Phytotherapy for chronic prostatitis. AB - Chronic prostatitis is a poorly defined condition that is difficult to treat; there are therefore multiple therapies. Although there is a paucity of trials using phytotherapeutic agents, plant extracts have been postulated to have anti inflammatory effects that might be useful in the treatment of chronic prostatitis. More placebo-controlled trials of longer duration in this condition are needed to ascertain whether there is a significant benefit to the use of phytotherapeutic agents for chronic prostatitis. PMID- 12084333 TI - Current Urology Reports--what makes us different? PMID- 12084334 TI - Neoadjuvant hormone therapy before radical prostatectomy does not improve disease specific survival. PMID- 12084335 TI - New insights and candidate genes and their implications for care of patients with hereditary prostate cancer. AB - A positive family history is a risk factor for prostate cancer. Most studies based on segregation analysis suggest autosomal dominant transmission of susceptibility genes. Multiple loci on chromosome 1 and chromosome X have been associated with prostate cancer by linkage analysis. The candidate gene approach has also revealed multiple genetic markers that are associated with increased risk for the disease. The genetic studies in prostate cancer suggest there are multiple genes involved in the development and progression of prostate cancer. PMID- 12084336 TI - The role of prostate-specific antigen velocity in prostate cancer early detection. AB - Prostate-specific antigen velocity (PSAV) is the rate of change in prostate specific antigen (PSA) values with repeated measurement over time. Accurate use of PSAV for prostate cancer early detection requires the use of two or more PSA levels collected over approximately 1.5 to 2 years. When these specimen collection criteria are met, more than 95% of men without prostate cancer will have a PSAV less than 0.75 ng/mL/y, whereas approximately 70% of men with prostate cancer will have a PSAV above this threshold. PSAV is thus more specific than routine PSA testing for the presence of prostate cancer, because few men (< 5%) without prostate cancer have a PSAV sufficient to trigger prostate biopsy. The use of PSAV in the increasing number of men with lengthy PSA histories obtained in systematic efforts at prostate cancer early detection may aid in diagnosing prostate cancer and spare some men unnecessary prostate biopsy. This review briefly summarizes the theoretic basis and clinical utility of PSAV in prostate cancer early detection. PMID- 12084337 TI - Continence-preserving anatomic radical retropubic prostatectomy: the "No-Touch" technique. AB - The technique of continence-preserving anatomic radical retropubic prostatectomy focuses on the preservation of the anatomic components of the external striated urethral sphincteric (EUS) complex. Three steps comprise this procedure for improving urinary control: 1) preservation of the anterolateral fascial attachments of the EUS, 2) protection of the innervation and the posterior fascial attachments of the EUS, and 3) avoidance of urethrovesical anastomotic sutures at the 5 and 7 o'clock positions. Preserving the external striated urethral sphincter and its innervation by performing a continence-preserving anatomic retropubic prostatectomy fascilitates a shorter time period for achieving urinary continence. PMID- 12084339 TI - Laparoscopic radical prostatectomy. AB - The radical prostatectomy has been modified over the years. With the introduction to the modern operating room of robots and other tools has come the latest modification: the laparoscopic radical prostatectomy (LRP), first described almost 10 years ago. In the past 2 years, the technique of LRP has been made standard, reproducible, and efficient. The LRP virtually eliminates the physical and emotional toll of radical prostate surgery and reduces blood loss, hospital time, and cost. Published series demonstrate oncologic and functional results comparable with and perhaps better than what is seen with open radical prostatectomy. The LRP is a standard surgical technique in a growing number of medical centers, with world experience estimated at 500 cases. The LRP offers the urologic community a rare opportunity for tangibly improving care. PMID- 12084338 TI - What does prostate-specific antigen recurrence mean? AB - The return of the prostate-specific antigen (PSA) to a detectable serum level (PSA recurrence) is usually the first sign of recurrent disease after radical prostatectomy. PSA recurrence generally occurs in men who are otherwise asymptomatic and may occur as late as 5 to 10 years after surgery. Men in this situation want to know what this means regarding the likelihood of clinical disease recurrence and, ultimately, survival. An evaluation for recurrent disease is warranted but generally does not reveal objective signs of clinical disease in the majority of men. Although select men may benefit from salvage local therapy, a PSA recurrence is most often an early sign of distant disease present since the time of surgery. The decision whether or not to initiate systemic therapy in these men is difficult and controversial. Fortunately, recent developments in determining the significance of a PSA recurrence may help the patient and his physician to make a more informed decision regarding treatment options. PMID- 12084340 TI - Cryosurgery for prostate cancer: new technology and indications. AB - Patients diagnosed with prostate cancer who elect to pursue active treatment of their disease must choose among the many available treatment alternatives. Several treatment options now exist for similar-stage disease (clinical T1 3N0M0), including radical prostatectomy, external beam radiation, prostate brachytherapy (PB), and cryosurgical ablation of the prostate (CSAP). This article reviews the current role of CSAP in the treatment of clinically localized prostate cancer. CSAP has a role in the primary treatment of men with high-risk, clinically localized prostate cancer (defined as PSA >10, Gleason score >or=7, or clinical stage >or= cT2B). CSAP (occasionally followed by external beam radiotherapy) appears to offer improved rates of cancer control over other types of single or combination therapies for this high-risk prostate cancer, and it is associated with an acceptable side-effect profile. CSAP should also be the treatment of choice for men with recurrent local disease who have undergone external beam radiotherapy or PB. PMID- 12084341 TI - Recent advances in chemotherapy for advanced prostate cancer. AB - Recently, several important studies have validated prostate-specific antigen (PSA) as a reliable measure of response to chemotherapeutic treatment in advanced hormone-refractory prostate cancer. Furthermore, although chemotherapy in this setting has always been considered palliative, several analyses of recent clinical trials have demonstrated a significant association between declines in PSA values of 50% or more and prolonged survival. Mitoxantrone, in combination with prednisone, has been shown to provide significant palliation and improved quality of life. The use of combinations of chemotheraputic agents also seems to provide significantly superior objective and subjective responses compared with single-agent regimens. In particular, estramustine has been shown to synergize many of the agents used in prostate cancer treatment and has been demonstrated to provide significant palliation and decline in PSA levels in combination with vinblastine, vinorelbine, etoposide, paclitaxel, and docetaxel. The results of several important trials of the taxanes both as single agents and in combination with estramustine have been completed in the past year and have demonstrated that these agents are very effective in the treatment of hormone-refractory prostate cancer. PMID- 12084342 TI - Screening for prostate cancer in African Americans. AB - African American men are known to have a higher risk of developing prostate cancer. Historically, African American men have presented at a higher stage and had a worse outcome from the disease than non-African American men. There is an ongoing debate whether this disparity is due to biologic, environmental, or behavioral factors, or a combination of these factors. Furthermore, lack of access to care is implicated. Despite this debate, there is emerging data that African American men and their families are receptive to education and early detection. Encouraging data from the military, Veteran's Administration, and private sector suggest that African American men can have a similar outcome to non-African American men if diagnosed early and treated effectively. Early detection efforts depend on prostate-specific antigen (PSA) testing. This article discusses various options for using the PSA test to more effectively screen African American men. In general, testing starting at age 40 is recommended using an upper limit of normal for PSA at 2.0 to 2.5 ng/mL for men between 40 and 49 years of age. In older men, maintaining this lower PSA threshold is reasonable to optimize curable cancer; however, published guidelines of 0 to 4.0, 0 to 4.5, and 0 to 5.5 ng/mL in African American men in their 50s, 60s, and 70s, respectively, are also recognized to balance the sensitivity and specificity of testing. Population-based prospective clinical trials of African American men are needed to further fine-tune the use of PSA in early detection, and to assess whether screening will improve the disease-specific mortality of prostate cancer in the population. PMID- 12084343 TI - Prostatic intraepithelial neoplasia. AB - High-grade prostatic intraepithelial neoplasia (PIN) is now accepted as the most likely pre-invasive stage of adenocarcinoma, a decade after its first formal description. PIN has a high predictive value as a marker for adenocarcinoma, and its identification warrants repeat biopsy for concurrent or subsequent invasive carcinoma. The only method of detection is biopsy; PIN does not significantly elevate serum prostate-specific antigen concentration or its derivatives and cannot be detected by ultrasound. Most studies suggest that most patients with PIN will develop carcinoma within 10 years. PIN is associated with progressive abnormalities of phenotype and genotype that are similar to cancer rather than normal prostatic epithelium, indicating impairment of cell differentiation with advancing stages of prostatic carcinogenesis. Androgen deprivation therapy decreases the prevalence and extent of PIN, suggesting that this form of treatment may play a role in chemoprevention. PMID- 12084344 TI - Quality of life following prostate cancer treatments. AB - Quality of life is of major concern to patients when choosing a treatment for prostate cancer. Health-related quality of life (HRQOL) is a patient-centered variable from the field of health services research that can be assessed in a valid and reliable manner. Using standardized questionnaires specifically designed to measure HRQOL in men with prostate cancer, we can now study the effect of various treatments on patients' quality of life. Treatments for metastatic prostate cancer can have significant effects in all areas of patients' quality of life. Patients with localized disease undergoing radical prostatectomy (RP) tend to have more sexual and urinary dysfunction than do men undergoing external beam radiation therapy (EBRT), although both groups have worse quality of life in these areas than age-matched controls. Men undergoing EBRT have worse bowel function than age-matched controls or men undergoing RP. Recent studies of men undergoing interstitial brachytherapy indicate that these patients have less urinary leakage than those who undergo RP, but experience considerably more irritative voiding symptoms, which can profoundly affect quality of life. Patients need to be informed of the possible impact of therapy on quality of life when choosing treatment. PMID- 12084345 TI - The role of free prostate-specific antigen in prostate cancer detection. AB - Prostate-specific antigen (PSA) is the most important serum tumor marker for prostate cancer detection. Free PSA is one of the many molecular forms of PSA that have been identified. Percent free PSA improves the specificity (elimination of unnecessary biopsies) for prostate cancer detection in men with nonsuspicious digital prostate examination and total serum PSA ranges between 4 and 10 ng/mL. Further study is necessary to determine the optimal clinical utility of percent free PSA in men with a total serum PSA level of less than 4 ng/mL. In addition, the level of free PSA may be affected by many factors, including age, prostate volume, prostate manipulation, sample handling, and type of assay used. PMID- 12084346 TI - Medulloblastomas and primitive neuroectodermal tumors rarely contain polyomavirus DNA sequences. AB - To address the hypothesis that medulloblastoma or supratentorial primitive neuroectodermal tumor (sPNET) can arise through infection by polyomaviruses, we examined genomic DNA isolated from 15 primary medulloblastoma and 5 sPNET biopsy specimens and from 2 medulloblastoma cell lines for the presence of DNA sequences from the polyomaviruses simian virus 40 (SV40), JC virus, and BK virus. These polyomaviruses have oncogenic potential in animals, and their DNA sequences have been detected in other surveys of various solid tumors, including childhood brain tumors. The tumor DNA samples were analyzed by Southern blot hybridization of polymerase chain reaction products that employed probes designed to detect specific polyomavirus sequences. Neither JC virus nor BK virus DNA sequences were detected in any of the specimens. None of the primary medulloblastoma or sPNET specimens contained SV40 sequences. However, SV40 DNA coding and noncoding sequences were detected in the D283-Med (medulloblastoma) cell line. Immunocytochemical studies of D283-Med revealed nuclear expression of SV40 large T antigen. In contrast to childhood ependymomas and choroid plexus tumors, medulloblastomas and sPNETs infrequently express evidence of polyomavirus infection. PMID- 12084347 TI - Cytoplasmic localization of wild-type p53 in glioblastomas correlates with expression of vimentin and glial fibrillary acidic protein. AB - Cytoplasmic accumulation of wild-type p53 in tumor cells indicates that the tumor suppressor is inactive with regard to growth suppressive functions. Whether this occurs randomly during tumor development or characterizes a certain tumor cell subset is not known. Here we assayed primary glioblastomas for expression and subcellular localization of p53 and determined a correlation with expression of intermediate filament proteins characterizing glial cell development. Sixty-nine percent of the tumors were p53 positive in immunohistochemistry. A significant number of tumors (23%) accumulated wild-type p53 in the cytoplasm, which correlated with the presence of vimentin and glial fibrillary acidic protein, except for 1 case. Tumors with exclusive nuclear p53 contained none or only one of these intermediate filament proteins. In an alternative approach, tumors positive for glial fibrillary acidic protein were screened for expression of p53 and vimentin. Thirty-eight percent of these tumors showed cytoplasmic p53, and all of those also expressed vimentin. Tumors with only nuclear p53 were vimentin negative, except for 1 case. No mutation was detected in p53 exons 5 to 8 in tumors with cytoplasmic p53, suggesting that they express wild-type p53. The data indicate that a cytoplasmic accumulation of wild-type p53 in human primary glioblastomas correlates with a certain intermediate filament protein expression, suggesting that it identifies a certain subset of tumors. PMID- 12084348 TI - Prognostic significance of an apoptotic index and apoptosis/proliferation ratio for patients with high-grade astrocytomas. AB - We evaluated the association of spontaneous apoptosis and an apoptosis/proliferation index with survival to determine the potential of such measures to serve as predictive markers for patients with glioblastoma multiforme (GBM). We examined the extent of spontaneous apoptosis in tumors from newly diagnosed patients, 75 with GBM and 21 with anaplastic astrocytoma, who were entered on treatment protocols of the North Central Cancer Treatment Group. In the group of GBM patients, those with a higher apoptotic index tended to live longer ( P = 0.04; Cox proportional hazards model including performance score, age, and extent of resection in a multivariate model). We found that the apoptotic index values for anaplastic astrocytoma patients tended to be lower than those in the GBM patients, although with small sample sizes, the result was not statistically significant ( P = 0.1). We also examined expression of the Ki 67 cell proliferation antigen immunohistochemically using the MIB-1 monoclonal antibody. Ki-67 expression did not provide additional information regarding the survival of patients with GBM. In this group of GBM patients, those patients with higher apoptotic index/proliferation ratios had a better prognosis than did those with a low ratio ( P < 0.021, same model as above). These findings suggest that both apoptosis and a cell death/cell proliferation ratio are associated with patient survival, and they may be useful for either the clinical evaluation of patients with GBM or the stratification of patients for treatment evaluation. PMID- 12084349 TI - Intravascular lymphomatosis presenting with a conus medullaris syndrome mimicking disseminated encephalomyelitis. AB - We describe the clinical, radiologic, and postmortem findings of a 42-year-old man with intravascular lymphomatosis. The patient presented with a conus medullaris syndrome followed by progressive, disseminated spinal and cerebral symptoms. Disseminated encephalomyelitis was suspected due to the clinical, radiologic, and cerebrospinal fluid findings and the results of a stereotactic brain biopsy, all of which were compatible with inflammatory CNS disease. Treatment with methylprednisolone and cyclophosphamide led to a temporary remission of symptoms. The patient died 13 months after onset of symptoms. The diagnosis of disseminated intravascular lymphomatosis was established after death. PMID- 12084350 TI - Paraneoplastic chorea: case study with autopsy confirmation. AB - A 67-year-old man presented with a 7-month history of insidiously progressive chorea, ataxia, and vertigo. Neurologic examination revealed deficits referable to the basal nuclei, cerebellar vermis, and vestibular nuclei. Small-cell lung cancer was diagnosed by fine-needle biopsy of a parahilar mass. After chemotherapy, the patient's chorea worsened. Anti-Hu antibodies were present in serum and cerebrospinal fluid. Microscopic examination of the brain at autopsy revealed diffuse perivascular lymphocytic infiltrates, microglial activation, and neuronophagia throughout the neuraxis, including the brainstem, cerebellum, lenticular nuclei, striatum, and cerebral cortex. Prominent loss of Purkinje cells was seen in the cerebellar vermis and hemispheres to a lesser degree. Chorea is extremely rare as a paraneoplastic manifestation of cancer. The florid presentation and the positive findings contrasted with an unremarkable MRI of the brain. This case illustrates the preeminence of symptoms and signs over negative MRI findings in paraneoplastic encephalitis. PMID- 12084353 TI - On antiretroviral guidelines and evidence based medicine. PMID- 12084354 TI - [Recommendations of the Spanish AIDS Study Group (GESIDA) and the National Aids Plan (PNS) for antiretroviral treatment in adult patients with human immunodeficiency virus infection in 2002]. AB - OBJECTIVE: To provide an update of recommendation on antiretroviral treatment (ART) in HIV-infected adults.Methods. These recommendations have been agreed by consensus by a committee of the spanish AIDS Study Group (GESIDA) and the National AIDS Plan. To do so, advances in the physiopathology of AIDS and the results on efficacy and safety in clinical trials, cohort and pharmacokinetics studies published in biomedical journals or presented at congresses in the last few years have been reviewed. Three levels of evidence have been defined according to the data source: randomized studies (level A), case-control or cohort studies (level B) and expert opinion (level C). Whether to recommend, consider, or not to recommend ART has been established for each situation. RESULTS: Currently, ART with combinations of at least three drugs constitutes the treatment of choice in chronic HIV infection. In patients with symptomatic HIV infection, initiation of ART is recommended. In asymptomatic patients initiation of ART should be based on the CD41/mL lymphocyte count and on the plasma viral load (PVL): a) in patients with CD41 lymphocytes < 200 cells/mL, initiation of ART is recommended; b) in patients with CD41 lymphocytes between 200 and 300 cells/mL, initiation of ART should, in most cases, be recommended; however, it could be delayed when the CD41 lymphocyte count remains close to 350 cells/mL and the PVL is low, and c) in patients with CD41 lymphocytes > 350 cells/mL, initiation of ART can be delayed. The aim of ART is to achieve an undetectable PVL. Adherence to ART plays a role in the durability of the antiviral response. Because of the development of cross-resistance, the therapeutic options in treatment failure are limited. In these cases, genotypic analysis is useful. Toxicity limits ART. The criteria for ART in acute infection, pregnancy and postexposure prophylaxis and in the management of coinfection with HIV and hepatitis C and B virus are controversial. CONCLUSIONS: The current approach to initiating ART is more conservative than in previous recommendations. In asymptomatic patients, the CD41 lymphocyte count is the most important reference factor for initiating ART. Because of the considerable number of drugs available, more sensitive monitoring methods (PVL) and the possibility of determining resistance, therapeutic strategies have become much more individualized. PMID- 12084351 TI - Pten signaling in gliomas. AB - In 1997, the PTEN gene (phosphatase and tensin homolog deleted on chromosome 10) was identified as a tumor suppressor gene on the long arm of chromosome 10. Since then, important progress has been made with respect to the understanding of the role of the Pten protein in the normal development of the brain as well as in the molecular pathogenesis of human gliomas. This review summarizes the current state of the art concerning the involvement of aberrant Pten function in the development of different biologic features of malignant gliomas, such as loss of cell-cycle control and uncontrolled cell proliferation, escape from apoptosis, brain invasion, and aberrant neoangiogenesis. Most of the tumor-suppressive properties of Pten are dependent on its lipid phosphatase activity, which inhibits the phosphatidylinositol-3'-kinase (PI3K)/Akt signaling pathway through dephosphorylation of phosphatidylinositol-(3,4,5)-triphosphate. The additional function of Pten as a dual-specificity protein phosphatase may also play a role in glioma pathogenesis. Besides the wealth of data elucidating the functional roles of Pten, recent studies suggest a diagnostic significance of PTEN gene alterations as a molecular marker for poor prognosis in anaplastic astrocytomas and anaplastic oligodendrogliomas. Furthermore, the possibility of selective targeting of PTEN mutant tumor cells by specific pharmacologic inhibitors of members of the Pten/PI3K/Akt pathway opens up new perspectives for a targeted molecular therapy of malignant gliomas. PMID- 12084355 TI - [Interpretative reading of the non-fermenting gram-negative bacilli antibiogram]. AB - Among non-fermenting Gram-negative rods, the most clinically important species are Pseudomonas aeruginosa, Acinetobacter baumannii, and Stenotrophomonas maltophilia, which are frequently multiresistant. P. aeruginosa resistance to beta-lactams depends on the production of chromosomal and plasmid-mediated beta lactamases, altered permeability (loss of OprD porin is related to carbapenem resistance) and active efflux pumps, particularly MexAB-OprM. In aminoglycoside resistant strains the main mechanism of resistance is the production of inactivating enzymes; the efflux pump MexXY-OprM is also involved. Quinolone resistance in P. aeruginosa is related to changes in topoisomerases, altered permeability and efflux pumps. The mechanisms of resistance of A. baumannii have not been well characterized, which makes interpretative reading of the antibiogram in this organism difficult. Resistance to beta-lactams is associated with the production of beta-lactamases and altered penicillin-binding proteins. Resistance to aminoglycosides has been related to modifying enzymes and resistance to quinolones to altered targets. S. maltophilia is resistant to carbapenems and other beta-lactams because of the production of two beta lactamases (L-1 and L-2). Aminoglycoside-modifying enzymes have also been described in this species. In contrast to what is observed in other organisms, S. maltophilia resistance to quinolones has been mainly related to active efflux, rather than to target alterations. PMID- 12084356 TI - [Neurobiology of narcolepsy]. PMID- 12084357 TI - [Weight gain after functional surgery for Parkinsons disease]. AB - BACKGROUND: The origin of weight gain after functional surgery for Parkinson's disease (PD) is incompletely known. We have done a prospective study to determine the possible causes of weight gain after pallidal and subthalamic surgery. PATIENTS AND METHOD: Twenty-seven patients were studied (9 unilateral pallidotomy, 9 bilateral deep brain stimulation (DBS) of palidum, and 9 bilateral DBS of suthalamic nucleus) with a follow-up of 12 months. The relationship between weight gain and changes in motor situation, levodopa dosage, dyskinesias, dysphagia and mood state were analyzed. The patients filled a questionnaire about the severity and etiology of weight gain. RESULTS: Weight gain was noted in 26 patients (mean of 4.7 kg at 12 months). It was found a significant correlation between weight gain and improvement of dyskinesias (AIMS) (r = 0.461; p = 0.023), the scores of the UPDRS part III, (r = 0.479; p = 0.028), and a significant inverse correlation with the pre-operative weight of the patient (r = 0.399; p = 0.050). Weight gain was most pronounced with bilateral than unilateral pallidal surgery (p = 0.021). The majority of patients referred weight gain as an slight adverse event and secondary to the improvement of dyskinesias. CONCLUSION: Functional surgery for PD, independently of the surgical target, provokes weight gain which is benign in the majority of cases. Reduction of energy expenditure with respect to the pre-operative situation would be the responsible of that phenomenon. PMID- 12084358 TI - [Neuropsychiatric inventory questionnaire (NPI-Q): Spanish validation of an abridged form of the Neuropsychiatric Inventory (NPI)]. AB - BACKGROUND: The Neuropsychiatric Inventory (NPI) is a validated clinical instrument for the evaluation of psychopathology in dementia. OBJECTIVE: To validate a brief questionnaire form of the NPI (NPI-Q) in Spanish from NPI-Q original version, intended for use in routine clinical practice. PATIENTS AND METHOD: We have developed a crossed validated form between NPI and NPI-Q in 120 Alzheimer's disease patients. RESULTS: Test-retest reliability of the NPI-Q, using Pearson correlation index was r = 0.89 for total symptom scale and r = 0.90 for distress scale. The prevalence of analogous symptom ratings differed by less than 6.7%. Convergent validity between NPI-Q and NPI, using Pearson correlation index was r = 0.879 for total symptom and r = 0.92 for distress scale. CONCLUSIONS: The NPI-Q Spanish version offers the possibility to use a reliable and brief instrument that can be used as a screening in the evaluation of neuropsychiatric symptoms in dementia and associated caregiver distress. It may be suitable for use in general clinical practice and it could be used as a brief neuropsychiatric interview. PMID- 12084360 TI - [Influence of infection related factors on neuropsychological performance of HIV seropositive subjects]. AB - There are no definitive risk factors which predict the development of neuropsychological deficits associated to HIV infection, and, taking into account that neuropsychological impairment is a risk factor of early death in all stages of HIV infection, there is a pressing need to determine those factors, either related or foreign to HIV, which help to explain the existing intragroup differences and that may also be associated with the development and progression of the dysfunction. The aim of this work is to revise those factors related with the infection that can influence neuropsychological functioning of the seropositive patients, such as antiretroviral treatments, immunological impairment, viral load and the advance of the infection. Identification of these risk factors will help us to outline characteristics of the seropositive subjects who are neuropsychologically more vulnerable, in order to detect them early with a view to preventing the manifestation of neuropsychological disturbances, and offer quick, suitable treatment. PMID- 12084359 TI - [Miguel Gayarre (1866-1936) and neurology in Madrid: a contribution to his biography]. AB - Miguel Gayarre y Espinal (1886-1936) is considered to be, together with Achucarro, one of the most scientifically influential persons on Lafora, and hence on the so-called Madrid Neurological School preceding the Spanish Civil War (late XIX and first third of the XX century). There is, however, a great deal of scarcity about personal biographic data on Gayarre. We document herein that Gayarre, after graduation, achieved his doctoral degree at the age of 29 by discoursing on a non-neurological subject (On essential paroxysmal tachycardia). It was only afterwards when Gayarre decided to become a pupil of Oppenheim, considered at the time to be a "pure" neurologist, in Berlin. His return to Spain might had been prompted following rejection to Oppenheim on the basis of creed and race to take the chair at La Charite Hospital after Westphal death. Although Lafora considered him "an excellent neurologist", Gayarre, as most other members of the Madrid Neurological School, combined his practice with Neurohistopathology as a student of Achucarro, and with Psychiatry, to become Director of the Ciempozuelos Hospital for the Insane in Madrid. PMID- 12084361 TI - [Bilateral carotid infarct]. PMID- 12084362 TI - [Room tilt illusion: Report of two cases and terminological review]. AB - The room tilt illusion is a transient misperception of the visual image as tilted on its side or even upside down; in this case it has been termed acute upside down reversal of vision. We report on two cases of room tilt illusion as manifestation of VIII nerve neuritis (herpes-zoster infection) and cerebellar hemorrhage. Room tilt illusion has been reported in association with vertebrobasilar stroke, migraine, multiple sclerosis, epilepsy and labyrinthine disorders. The pathophysiology of this rare visual illusion has been related to a lesion of the visual or vestibulo-otolith pathways. In animals the neurones of the parieto-insular vestibular cortex areas are multisensory. So, they can respond to somatosensory, optokinetic and visual stimuli. In humans the knowledge about vestibular cortex function and localization is less precise than in animals. However, we propose a disorder of multisensorial vestibular cortex, resulting from a lession of vestibular pathways or association cortex, as mechanism of this phenomenon. PMID- 12084363 TI - [Chronic subdural hematoma simulating a TIA. Implications for the management transient neurological deficit]. AB - Chronic subdural hematoma that generally happens after cranial trauma doesn't have clinical manifestations until days or weeks after the traumatism. Due to the lesions nature, symptoms are almost always progressive and presentation as transient ischemic attacks (TIA) is very uncommon. We describe 2 cases of chronic subdural hematoma that started simulating TIA and we discuss its implications in the management of those patients who had presented transient neurological deficit. PMID- 12084364 TI - [Central nervous system vasculitis-associated Antons syndrome]. PMID- 12084366 TI - [Prophylaxis of intracranial hypotension syndrome and headache following lumbar puncture]. PMID- 12084368 TI - [Effect of antihypertensive therapy on pulse pressure]. AB - BACKGROUND: Pulse pressure (difference between systolic and diastolic pressure) is an important prognostic factor for cardiovascular mortality and morbidity in elderly hypertensives. However, data regarding the effect of antihypertensive treatment on pulse pressure (PP) are scarce. In the present study, we evaluate the effect of six classes of antihypertensive drugs on PP in an elderly hypertensive cohort. PATIENTS AND METHOD: It was an open, prospective and multicenter study performed by primary care physicians. 857 hypertensive patients (54% women) with a mean age of 68 years were included. Antihypertensive treatment (any antihypertensive drug used in monotherapy) was freely assigned by investigators and then grouped in classes for analysis. Blood pressure was measured by a validated oscillometric device using a standardized protocol. RESULTS: ACE inhibitors were the mostly used class of antihypertensive drugs (27.8%). We found no differences between drug classes in PP reduction. Likewise, no differences were observed regarding the effect on systolic, diastolic, and mean blood pressure. The percentage of adverse reactions was low (6.3%). When effects on PP reduction and adverse reactions were pooled together, angiotensin receptor blockers emerged as the antihypertensive drug class with the best profile. CONCLUSIONS: Antihypertensive drugs do not differ substantially in their ability to reduce PP. Although PP is considered as an important prognostic factor for cardiovascular mortality and morbidity, the present results do not strengthen its usefulness as a distinctive marker of antihypertensive drug classes. PMID- 12084369 TI - [Relative role of physical inactivity and snacking between meals in weight gain]. AB - BACKGROUND: Diet and life-styles are considered as the main factors that determine the high prevalence of obesity in Western societies. Although some countries have registered a decrease in fat intake, the percentage of overweight and obesity has increased. Therefore, it is thought that fat intake may not be the main factor that determines the current epidemic of obesity. The objective of this study was to determine the role of a sedentary life-style and eating between meals (snacking) as major determinants of a recent weight change (over last 5 years). METHOD: By using cross-sectionally baseline data of the SUN cohort, we adjusted non-conditional logistic regression models to estimate the odds ratio (OR) of gaining weight according to age, physical activity in leisure time, watching television, taking a nap, smoking, snacking and the intake of macronutrients. RESULTS: A statistically significant inverse association between leisure-time physical activity and the probability of gaining weight was found for men (OR = 0.93; CI 95%, 0.88-0.98) and a trend was also present among women. Snacking was positively associated with a higher probability of gaining weight among men (OR = 1.88; CI 95%, 1.40-2.53) and among women (OR = 1.38; CI 95%, 1.10 1.73). CONCLUSIONS: Our data suggest a direct association between snacking and weight gain in middle-aged people. PMID- 12084370 TI - [Effectiveness of a group therapy intervention to quit smoking. Randomized clinical trial]. AB - BACKGROUND: Group therapy is a widely used technique for the treatment of tobacco, alcohol and other toxic addictions. Nevertheless, its effectiveness for smoking cessation remains to be established. Our objective was to evaluate the relative effectiveness of a group intervention versus an individual intervention for smoking cessation. METHOD: Randomized clinical trial of 106 smokers wishing to quit their addiction, who were assigned to two types of intervention: Short Individual Intervention (SII) and Group Intervention (GI). Nicotine patches were administered as supplemental therapy when needed in both instances. Results were assessed by intention to treat analysis. RESULTS: Although smoking cessation rates decreased in parallel with longer follow-up periods in each intervention group (from 39.6 to 26.4% and from 22.6 to 15.1% for GI and SII groups, respectively, at 3 and 6-months), the relative effectiveness was similar during both periods and there was no better response to any intervention at 3-months (relative risk [RR] = 1.75; CI95%, 0.96-3.18) and at 6-months (RR = 1.75; CI95%, 0.80-3.82). Compliance with GI was low and less than 60% of smokers attended to 5 out of 7 GI sessions. CONCLUSIONS: Group intervention (GI) is not more effective than short individual intervention to quit smoking. However, this fact could be due to the low compliance observed with regard to smokers' attendance to GI sessions. PMID- 12084371 TI - [Origins of hypertension: birth weight, a new issue]. PMID- 12084372 TI - [Agoraphobia]. PMID- 12084373 TI - [Edema in lower limbs and scrotum in a 66-year-old male]. PMID- 12084374 TI - [Prolonged out-of-hospital prophylaxis of venous thromboembolism]. PMID- 12084375 TI - [Where are you, odds ratio? Where are you, control group?]. PMID- 12084377 TI - [Serum sickness-like reaction induced by bupropion]. PMID- 12084378 TI - [Antiretroviral prophylaxis in a pregnant hospital worker]. PMID- 12084379 TI - [Desensitization to fluconazole in patient with human inmunodeficiency virus infection]. PMID- 12084380 TI - [Recurrent venous thromboembolism with increase of factor VIII and familial clustering]. PMID- 12084381 TI - [Crohns disease in a patient with Hashimotos thyroiditis and Sjogrens syndrome]. PMID- 12084382 TI - [Gastric tuberculosis: a condition to be distinguished from gastric sarcoidosis]. PMID- 12084383 TI - The low conductance K(+) channel in human colonic crypt cells has a voltage dependent permeability not affected by Mg(++). AB - The low conductance K(+) channel found in human colonocytes was investigated using the patch-clamp technique. The channel is Ca(++)-dependent and is blocked by Ba(++) (5 mM) with a decrease in open probability from 0.42 to 0.19. At -40 mV the slope conductance was 29 pS (using intracellular solution in the pipette). In inside-out patches, inward rectification was seen both with KCl (pipette)/NaCl (bath) solutions as well as KCl/KCl solutions. The rectification could not be affected by omitting Mg(++) from the pipette or the bath solution, neither by exposing the patches to the polyamine spermine (1 mM). Using the Goldman-Hodgkin Katz equation we show that the permeability decreased in a linear fashion from approximately 5.2 x 10(-14) cm(3)/s to 1.8 x 10(-14) cm(3)/s (-100 to +100 mV), both with and without Mg(++) in the solutions. There was no significant difference in the nominal values of permeability. This property of the K(+) channel may facilitate the hyperpolarization needed to sustain a chloride secretion. PMID- 12084384 TI - Inhibition of estrogenic stimulation of gene expression by genistein. AB - Two principle soy-derived isoflavones, genistein and daidzein, are believed to play a key role in inhibiting tumor growth. The molecular basis of the anti-tumor activity of these two isoflavones has not been fully established. To determine the mechanism of action of the above phytochemicals on estrogen-responsive genes, we tested the effect of the same on the expression of Estrogen-Regulated mRNA Stabilizing Factor (E-RmRNASF). E-RmRNASF is expressed in the liver in response to estrogen, and is responsible for estrogen-mediated stabilization of apolipoprotein II mRNA (Ratnasabapathy, 1995, Cell. Mol. Biol. Res, 41: 583-594). The estrogen-mediated hepatic expression of apolipoprotein II mRNA is regulated transcriptionally, and also post-transcriptionally in part by stabilization of its mRNA. E-RmRNASF protects the RNA from targeted endonucleolytic degradation. The hepatic expression of E-RmRNASF is also modulated by certain estrogenic and antiestrogenic nonsteroidal environmental xenobiotics (Ratnasabapathy et al., 1997, Biochem. Pharmacol., 53: 1425-1434). Roosters were administered estrogen, genistein, or daidzein parenterally and tested for hepatic expression of E RmRNASF. Expression of E-RmRNASF in the livers was stimulated in roosters who received estrogen and genistein, indicating that they are agonistic at the chicken estrogen receptor. However, a lack of induction of E-RmRNASF expression in the liver was seen with control roosters treated with the vehicle and those treated with daidzein. To determine whether daidzein was anti-estrogenic, roosters were given combinations of estrogen and increasing concentrations of daidzein. Daidzein at concentrations ranging from 5-1000 micromole/kg failed to antagonize stimulation of E-RmRNASF by 5 micromoles/kg estrogen. To determine whether genistein or daidzein exerted partial agonistic effects, roosters were given increasing concentrations of genistein, daidzein or estrogen alone, or combinations of estrogen and increasing doses of genistein or daidzein. At low to intermediate concentrations genistein by itself failed to stimulate E-RmRNASF, and was agonistic only at high concentrations. Genistein at the low concentrations failed to antagonize estrogenic stimulation of E-RmRNASF. At the intermediate concentrations however, genistein blocked stimulation of E-RmRNASF by estrogen, even though by itself could not exert a stimulatory effect. At the higher concentrations genistein stimulated E-RmRNASF regardless of the presence or absence of estrogen. At the higher ratios, the lack of inhibition of estrogenic stimulation by genistein was most likely due to its own agonistic activity. Therefore, genistein appears to behave as a partial agonist; behaves as an agonist by itself, and as an antagonist in the presence of estrogen. However, daidzein did not display any estrogenic or antiestrogenic activity at the concentrations tested. PMID- 12084385 TI - Prolonged impact of pubertal serotonin treatment (hormonal imprinting) on the later serotonin content of white blood cells. AB - The first encounter between the developing receptor and its target hormone establishes the hormonal imprinting which is needed for the normal function of the cell. In the presence of foreign-however able to bind-molecules, faulty imprinting develops with lifelong consequences. Hormonal imprinting influences not only the receptors, but also the later hormone production of cells. The critical time of hormonal imprinting is the perinatal period, however it can be executed sometimes (in continuously differentiating cells) also at puberty. As in earlier experiments single neonatal serotonin treatment caused a life-long alteration of white blood serotonin content in female rats, the early (10-19 day) and late (8 weeks) effect of single pubertal serotonin treatment was studied presently, by using flow cytometry. In contrast to the earlier (neonatal) results, pubertal treatment caused a radical reduction of serotonin content in male's lymphocytes, monocytes, granulocytes and mast cells, independent on the time of study. The effect in females was rather increasing, however uncertain. The experiments call attention to the possible different effects of neonatal and pubertal hormonal imprinting and to the imprintability of blood cells in adolescence. PMID- 12084386 TI - High glucuronidation activity of environmental estrogens in the carp (Cyprinus carpino) intestine. AB - Many adverse effects on carp reproductive organs have been reported to be caused by exposure to environmental estrogens, such as nonylphenol and bisphenol A, which contaminate the aquatic environment. The glucuronidation activities of xenoestrogens (bisphenol A and diethylstilbestrol) and phytoestrogens (coumestrol, genistein and biochanin A), but not nonylphenol and octylphenol, were observed in microsomes prepared from carp organs. The highest levels of glucuronidation of environmental estrogens, for which the optimum temperature was 25-30 degrees C, were observed in the intestinal microsomes of 2-year-old carp. These activities in carp intestine increased developmentally, and the maximum levels corresponded to 5-10 % of that in rat liver microsomes. However, the glucuronidation of phytoestrogen by carp intestinal microsomes corresponded to that of rat liver microsomes. Only bisphenol A-glucuronide was excreted from the everted intestine, indicating that bisphenol A is metabolized in the carp intestine mainly as glucuronide. These results suggest that glucuronidation by carp intestine plays an important role for the detoxification of xenoestrogens and phytoestrogens, except for nonylphenol and octylphenol. PMID- 12084387 TI - Involvement of angiotensin II, TRH and prolactin-releasing peptide in the estrogen-induced afternoon prolactin surge in female rats: studies using antisense technology. AB - The roles of endogenous angiotensin II (AII), thyrotropin-releasing hormone (TRH) and prolactin-releasing peptide (PrRP) on the estrogen-induced prolactin (PRL) surge and the diurnal change of tuberoinfundibular dopaminergic (TIDA) neuronal activity were assessed in this study. Ovariectomized, estrogen-primed rats implanted with intracerebroventricular cannula received daily injection of antisense oligodeoxynucleotide (ODN, 10 microg/3 microl) against the mRNA of AII, TRH or PrRP for two days. Artificial cerebrospinal fluid or the sense ODN were used as the control. In the first experiment, serial blood samples (0.3 ml each) were obtained hourly from each rat through a pre-implanted intraatrial catheter from 1100 to 1700h. Half of the rats pretreated with respective antisense ODN received single injections of AII, TRH or PrRP (1 microg each, i.v.) at 1400h. In the second experiment, groups of rats were decapitated either at 1000 or 1500h. The hypothalamic median eminence tissue of each rat was dissected out and its DOPAC content was used as the index for TIDA neuronal activity. Plasma and serum PRL levels were determined by radioimmunoassay. Pretreatment of antisense ODN against the mRNA of either AII or TRH significantly attenuated the PRL surge; replacement injection of AII or TRH restored the surge. The effect of antisense ODN against PrRP was less significant. None of the treatments significantly affected the diurnal changes of TIDA neuronal activity. In summary, both AII and TRH may play an important role as the PRL-releasing hormone involved in the estrogen-induced afternoon PRL surge. PMID- 12084388 TI - Effects of GABA on circular smooth muscle spontaneous activities of rat distal colon. AB - GABAergic regulation of intestinal motility through the modulation of non adrenergic non-cholinergic (NANC) neurons remains poorly understood especially in rat colon where very few studies have been undertaken. Therefore, the effects of GABA on circular preparations of rat distal colon were investigated using classical organ bath chambers to record spontaneous mechanical activities (SMA). SMA was characterized by the occurrence of rhythmic phasic contractions (type-I) or by spontaneously occurring large contractions superimposed on small rhythmic contractions (type-II). In the presence of atropine and guanethidine (NANC conditions), these large contractions were inhibited by bicuculline, a GABA(A) receptor antagonist as well as by TTX, L-NAME and apamin together, or L 732-138, a NK1-receptor antagonist. In NANC conditions, GABA induced a transient monophasic relaxation or a biphasic effect characterized by a relaxation followed by a tonic contraction in both type-I and -II preparations. Both the inhibitory and excitatory effects of GABA were blocked by TTX and L-NAME + apamin; the GABA induced contraction was also sensitive to L 732-138. The responses to GABA were mimicked by the GABA(A)-receptor agonist, muscimol, whereas baclofen and CACA, respectively GABA(B) and GABA(C)-receptors agonists showed no effect. These results demonstrated that only GABA(A)-receptors seem to be involved in the regulation of SMA in rat distal colon in NANC conditions. Release of NANC inhibitory transmitter (NO and probably ATP) and NANC excitatory transmitter (maybe substance P) might be involved. PMID- 12084389 TI - SSTR2 mediates the somatostatin-induced increase in intracellular Ca(2+) concentration and insulin secretion in the presence of arginine vasopressin in clonal beta-cell HIT-T15. AB - The effects of somatostatin (SRIF) are mediated through the seven transmembrane receptor family that signals via Gi/Go. To date, five distinct SRIF receptors have been characterized and designated SSTR1-5. We have characterized the SRIF receptor that mediates the increase in [Ca(2+)](i) and insulin secretion in HIT T15 cells (Simian virus 40-transformed Syrian hamster islets) using high affinity, subtype selective agonists for SSTR1 (L-797,591), SSTR2 (L-779,976), SSTR3 (L-796,778), SSTR4 (L-803,087), SSTR5 (L-817,818) and PRL-2903, a specific SSTR2 antagonist. In the presence of arginine vasopressin (AVP), SRIF increased [Ca(2+)](i) and insulin secretion. Treatment with the SSTR2 agonist L-779,976 resulted in similar responses to SRIF. In addition, L-779,976 increased both [Ca(2+)](i) and insulin secretion in a dose-dependent manner. Treatment with L 779,976 alone did not alter [Ca(2+)](i) or basal insulin secretion. In the presence of AVP, all other SRIF receptor agonists failed to increase [Ca(2+)](i) and insulin secretion. The effects of SRIF and L-779,976 were abolished by the SSTR2 antagonist PRL-2903. Our results suggest that the mechanism underlying SRIF induced insulin secretion in HIT-T15 cells be mediated through the SSTR2. PMID- 12084391 TI - Antipsychotic profile of 5-[2-[4-(6-fluoro-1H-indole-3-yl)piperidin-1-yl]ethyl]-4 (4-fluorophenyl)thiazole-2-carboxylic acid amide (NRA0562) in rats. AB - The antipsychotic profile of 5-[2-[4-(6-fluoro-1H-indole-3-yl)piperidin-1 yl]ethyl]-4-(4-fluorophenyl)thiazole-2-carboxylic acid amide (NRA0562) was investigated using the conditioned avoidance test in rats. NRA0562 is a putative "atypical" antipsychotic agent with moderate to high affinities for dopamine D(1), D(2), D(4), 5-hydroxytryptamine(2A) receptors and alpha(1) adrenoceptor. NRA0562 (1 and 3 mg/kg, p.o.) dose-dependently and significantly impaired the conditioned avoidance response. Likewise other atypical antipsychotics such as risperidone (1 and 3 mg/kg, p.o.) and clozapine (100 mg/kg, p.o.) dose dependently and significantly impaired the conditioned avoidance response in rats. In addition, typical antipsychotics, haloperidol (1 and 3 mg/kg, p.o.) potently impaired the conditioned avoidance response.These results suggest that antipsychotic profile of NRA0562 is consistent with profiles of clozapine or risperidone and may be considered an atypical antipsychotic agent. PMID- 12084390 TI - Effects of losartan on the blood-brain barrier permeability in long-term nitric oxide blockade-induced hypertensive rats. AB - Hypertension is closely associated with vascular endothelial dysfunction. The aim of this study was to investigate the effects of Angiotensin II (ANG II) receptor antagonist losartan on the blood-brain barrier (BBB) permeability in L-NAME induced hypertension and/or in ANG II-induced acute hypertension in normotensive and hypertensive rats. Systolic blood pressure was measured by tail cuff method before, during and following L-NAME treatment (1 g/L). Losartan (3 mg/kg) was given to the animal for five days. Acute hypertension was induced by ANG II (60 microg/kg). Arterial blood pressure was directly measured on the day of the experiment. BBB disruption was quantified according to the extravasation of the albumin-bound Evans blue dye. Losartan significantly reduced the mean arterial blood pressure from 169 +/- 3.9 mmHg to 82 +/- 2.9 mmHg in L-NAME and from 171 +/ 2.9 mmHg to 84 +/- 2.9 in L-NAME plus losartan plus ANG II groups (p < 0.05). The content of Evans blue dye in the cerebral cortex significantly increased in L NAME (p < 0.01). Moreover, the content of Evans blue dye markedly increased in the cerebellum (p < 0.001) and slightly increased in diencephalon region (p < 0.05) in L-NAME plus ANG II. Losartan reduced the increased BBB permeability to Evans blue dye in L-NAME (p < 0.01) and L-NAME plus ANG II (p < 0.001). These results indicate that L-NAME and L-NAME plus ANG II both lead to an increase in microvascular Evans blue dye efflux to brain, and losartan treatment attenuates this protein-bound dye transport into brain tissue presumably due to its protective effect on endothelial cells of brain vessels. PMID- 12084392 TI - Effects of chronic methamphetamine exposure on heart function in uninfected and retrovirus-infected mice. AB - Methamphetamine (MA) increases catecholamine levels, which have detrimental effects on heart function through vasoconstriction, myocardial hypertrophy, and fibrosis. Murine retrovirus infection induces dilated cardiomyopathy (DCM). The present study investigated the cardiovascular effects of chronic MA treatment on uninfected and retrovirus-infected mice. C57BL/6 mice were studied after 12 weeks treatment. The four study groups were (group I) uninfected, MA placebo; (group II) infected, MA placebo; (group III) uninfected, MA treatment; and (group IV) infected and MA treatment. MA injections were given i.p. once a day for 5 days/week with a increasing dose from 15 mg/kg to 40 mg/kg. Left ventricular mechanics were measured in situ a using Millar conductance catheter system for pressure-volume loop analysis. Cardiac pathology was determined with histological analysis. In the uninfected mice, the load independent contractile parameters, pre-load recruitable stroke work (PRSW) and dP/dt(max) vs. Ved, significantly decreased by 32% and 35% in MA treated mice when compared to the saline injected mice. In retrovirus-infected mice, although there were no significant difference in Ees, PRSW, and dP/dt(max) vs. Ved due to MA treatment, they were increased 45%, 15% and 42% respectively when compared to saline treated mice. No further lowered heart function during murine AIDS may be due to the counteraction of the retroviral DCM and the MA induced myocardial fibrosis and hypertrophy (thickening of the ventricular walls). This is supported by increases in the End-diastolic volume (Ved, 38%) and End-systolic volume (Ves, 84%) in the retrovirus-infected saline injected mice, the decreases of 33% and 17% in the uninfected MA-treated mice, but no significant changes in the retrovirus-infected MA treated mice when compared to uninfected saline injected mice. These data suggest that MA induced myocardial cellular changes compensate for retrovirus induced DCM. PMID- 12084393 TI - Zidovudine (AZT) treatment suppresses granulocyte-monocyte colony stimulating factor receptor type alpha (GM-CSFR alpha) gene expression in murine bone marrow cells. AB - In vitro exposure of murine bone marrow cells to increasing concentrations of zidovudine (AZT, 0.1-50 microM) had a concentration dependent suppressive effect on the growth of granulocyte-monocyte colony forming unit (CFU-GM) derived colonies. In our previous published study, the mechanism of AZT-induced suppression of erythroid colony forming unit (CFU-E) derived colonies was linked to a decrease in erythropoitin receptor (Epo-R) gene expression. In this study, we have observed that AZT exposure also induced a concentration dependent suppressive effect (35-90%) on GM-CSF receptor type alpha (GM-CSFR alpha) gene expression. The suppression of GM-CSFR alpha mRNA expression was specific, since AZT caused a much lower decrease (15-22%) on the IL-3 receptor type alpha (IL-3R alpha) message level, and had an insignificant effect on glyceraldehyde 3 phosphate dehydrogenase (GAPDH) and c-myc message levels. Erythropoietin (Epo) therapy has been used for reversal of AZT induced erythroid toxicity. Exposure to increasing concentrations (10-500 U/ml) of GM-CSF was unable to override the suppressive effect of AZT on CFU-GM derived colonies, however, treatment in combination with IL-3 (10-250 U/ml) ameliorated the suppressive effects of AZT on CFU-GM and on GM-CSFR alpha and IL-3R alpha gene expression. These findings suggest a mechanism via which AZT may suppress granulocyte-monocyte specific differentiation in murine bone marrow cells. These data also suggest that a combination of GM-CSF and IL-3 may be a superior therapeutic intervention for AZT induced neutropenia. PMID- 12084394 TI - Introduction: the role of hormone replacement therapy in prevention and treatment of cardiovascular disease in postmenopausal women. PMID- 12084395 TI - Cardiovascular disease in postmenopausal women: myths and reality. AB - Although cardiovascular disease (CVD) has traditionally been considered a disease that affects middle-aged men, it also has a profound effect on women. By age 65, the number of deaths from CVD in women surpasses deaths in men by 11%. Cardiovascular disease is the leading cause of mortality in women. Despite the impact of CVD, women as a group, as well as healthcare professionals, have not focused on this disease entity. As a result, women may not make adjustments that could reduce their risk for CVD, and healthcare professionals may not adequately counsel women on risk modification, which may include lifestyle changes and pharmaceutical intervention. Although there are many similarities, women differ from men in both disease presentation and prognosis for CVD. Because of the difference in presentation, diagnoses in women may be delayed, potentially causing further harm. Although the number of deaths caused by CVD has been decreasing in men, this trend has yet to be observed in women. PMID- 12084397 TI - Protective effects of estrogen on the cardiovascular system. AB - Estrogen has direct and indirect effects on the cardiovascular system that are mediated by the estrogen receptors ER-alpha and ER-beta. The direct effects of estrogen occur through rapid nongenomic and longer-term genomic pathways. The rapid effects of estrogen are mediated by ERs and result in the activation of endothelial nitric oxide synthase, leading to arterial vasodilation. Longer-term effects involve changes in gene and protein expression, modulating the response to injury and atherosclerosis. Estrogen also indirectly influences serum lipoprotein and triglyceride profiles, and the expression of coagulant and fibrinolytic proteins. Advanced atherosclerosis and certain progestins, however, may attenuate some of the protective effects of estrogen. PMID- 12084399 TI - Atherosclerosis imaging methods: assessing cardiovascular disease and evaluating the role of estrogen in the prevention of atherosclerosis. AB - Various interventions for cardiovascular disease (CVD) slow or reverse the progression of atherosclerosis and reduce the risk of clinical coronary events. Although the cardiovascular benefits of hormone replacement therapy have been demonstrated in observational studies in predominantly healthy women, no benefit has been found in a randomized clinical trial conducted in older women with established CVD. It is possible that the benefit of hormone therapy occurs when it is used relatively early in the progression of atherosclerosis. Techniques are now available to monitor the various stages of atherosclerosis. Quantitative coronary angiography, a technique used to evaluate relatively late-stage atherosclerosis, has been shown to predict the risk of subsequent clinical coronary events. B-mode ultrasonography of the intima-media wall thickness (IMT) of the carotid artery can assess the earlier stages of atherosclerosis and correlates with atherosclerosis risk factors, as well as with clinical cardiovascular and cerebrovascular outcome. This technique offers a relatively rapid and cost-effective method to test therapies for CVD and to screen for individuals who are at high risk for cardiovascular events. As an example of the use of atherosclerosis imaging to evaluate possible therapeutic interventions, measurements of IMT were performed in a randomized, controlled trial comparing oral 17beta-estradiol with placebo. The results demonstrated that 17beta estradiol significantly reduces the progression of subclinical atherosclerosis in healthy, postmenopausal women when compared with placebo. PMID- 12084401 TI - Risk factors for coronary artery disease in women. AB - Because cardiovascular disease (CVD) is the most important cause of death in women in the United States, it is imperative that the main risk factors for CVD in women be identified and modified. The risk factors that have the strongest impact on the incidence of CVD in women are not necessarily the same as those for men. The risk for women increases at menopause, most likely because of the decrease in levels of circulating estrogen. The classic risk factor for CVD is altered lipid levels. In middle-aged women, elevated low-density lipoprotein cholesterol levels are somewhat less important relative to lowered levels of high density lipoprotein cholesterol and elevated triglyceride levels as independent risk factors. The metabolic syndrome, which encompasses a range of conditions known to be CVD risk factors, also has a greater impact on the incidence of CVD in women than in men. Various emerging risk factors appear to be important indicators for vascular disease in women, including C-reactive protein, homocysteine, and lipoprotein(a) levels. Many of these risk factors are affected by hormone replacement therapy, which may diminish CVD risk in postmenopausal women. Because of the complex origin of CVD, it is important to target the full array of risk factors for modification, rather than focusing on a single factor or treatment to the exclusion of other important markers. PMID- 12084403 TI - Hormone replacement and the prevention of cardiovascular disease. AB - Recently, there has been considerable discussion about the benefits and risks of using hormone replacement therapy (HRT) for cardiovascular protection in postmenopausal women. Numerous studies have addressed this issue, with conflicting results. Observational studies strongly suggest a benefit of HRT for primary prevention of cardiovascular disease (CVD), whereas controlled trials that tested a single common regimen of estrogen plus progestin found no effect on CVD events or angiographic progression in older women with established CVD. More recently a primary prevention trial in high risk patients found reduced carotid atheroma with estrogen alone. A variety of combination therapies with various estrogens and progestins are now available. These formulations have significantly different effects on cardiovascular risk markers, which independently affect outcomes. This information should be considered when deciding whether to use HRT in an individual woman, and if so, which formulation to use. PMID- 12084405 TI - Effects of progestins in different hormone replacement therapy formulations on estrogen-induced lipid changes in postmenopausal women. AB - Hormone replacement therapy (HRT), used by many women to relieve the symptoms of menopause and for the prevention of osteoporosis, is available in an increasing number of formulations. Options include various combinations of estrogen and progestin as well as different routes of administration. For women with a uterus, who require a progestin, a convenient and commonly used option is combining the estrogen and progestin in a single pill. Such formulations have the lipid modifying effects of estrogen, altered to specific type and dose of progestin used. The cardioprotective increases in levels of high-density lipoprotein cholesterol associated with exogenous estrogen may be blunted or even reversed with certain HRT regimens. In addition, some progestins minimize the increase in triglycerides that occurs with exogenous estrogen, whereas others do not. Because of the importance of lipid markers as risk factors for cardiovascular disease in postmenopausal women, these lipid effects should be carefully considered when selecting an HRT regimen. PMID- 12084407 TI - Somatostatin immunoreactivity in axon terminals in rat nucleus tractus solitarii arising from central nucleus of amygdala: coexistence with GABA and postsynaptic expression of sst2A receptor. AB - Axon terminals synapsing on neurones in the nucleus tractus solitarii (NTS) that originate from the central nucleus of the amygdala (CeA) have been shown to contain gamma-aminobutyric acid (GABA) immunoreactivity. Here we investigated whether such terminals also contain somatostatin (SOM), a neuropeptide found in axons distributed throughout the NTS and in somata in the CeA, and known to modulate cardiovascular reflexes when microinjected into the NTS. With fluorescence microscopy, SOM immunoreactivity was seen in the varicosities of some axons throughout the NTS that were anterogradely labelled with biotin dextran amine injected into the CeA. Such varicosities were frequently observed in close proximity to dendrites of NTS neurones that were immunoreactive for the SOM receptor sst(2A) subtype, and in many cases also for catecholamine synthesising enzymes. In the caudal, cardioregulatory zone of NTS, SOM immunoreactivity was localised by electron microscopic pre-embedding gold labelling to boutons containing dense-cored and clear pleomorphic vesicles and forming symmetrical synapses, mostly onto dendrites. Additional post-embedding gold labelling for GABA suggested that a subpopulation (29%) of GABAergic terminals sampled in this area of NTS contained SOM. Almost all boutons anterogradely labelled from the amygdala were GABA-immunoreactive (-IR) and 21% of these were SOM-IR. A similar proportion of these boutons (22%) formed synapses onto dendrites containing immunoreactivity for the SOM receptor sst(2A) subtype. These observations provide evidence that some of the GABAergic projection neurones in the CeA that inhibit baroreceptor reflex responses in the NTS in response to fear or emotional stimuli could release SOM, which might modulate the activity of NTS neurones via an action on sst(2A) receptors. PMID- 12084408 TI - The distribution of vasotocin and mesotocin immunoreactivity in the brain of the snake, Bothrops jararaca. AB - Polyclonal antibodies against vasotocin (AVT) and mesotocin (MST) were used to explore the distribution of these peptides in the brain of the snake Bothrops jararaca. Magnocellular AVT- and MST-immunoreactive (ir) perikarya were observed in the supraoptic nucleus (SON), being AVT-ir neurons more numerous. A portion of the SON, in the lateroventral margin of the diencephalon ventrally to optic tract, showed only AVT-ir perikarya and fibers. However, the caudal most portion displayed only mesotocinergic perikarya. Parvocellular and magnocellular AVT- and MST-ir perikarya were present in the paraventricular nucleus (PVN) being AVT-ir fibers more abundant than MST-ir. Vasotocinergic perikarya were also found in a dorsolateral aggregation (DLA) far from the PVN. Mesotocinergic perikarya were also present in the recessus infundibular nucleus and ependyma near to paraventricular organ. Nerve fibers emerging from supraoptic and paraventricular nuclei run along the diencephalic floor, internal zone of the median eminence (ME) to end in the neural lobe. Also a dense network of AVT- and MST-ir fibers was present in the external zone of the ME, close to the vessels of the hypophysial portal system. As a rule, all regions having vasotocinergic and mesotocinergic perikarya also showed immunoreactive fibers. Vasotocinergic and mesotocinergic fibers but not perikarya were found in the lamina terminalis (LT). Moreover AVT-ir fibers were present in the nucleus accumbens and MST-ir fibers in the septum. In mesencephalon and rhombencephalon MST-ir fibers were more numerous than AVT-ir fibers. Vasotocinergic and mesotocinergic fibers in extrahypothalamic areas suggest that these peptides could function as neurotransmitters and/or neuromodulators in the snake B. jararaca. PMID- 12084409 TI - The medial preoptic nucleus receives vasotocinergic inputs in male quail: a tract tracing and immunocytochemical study. AB - The sexually dimorphic testosterone-sensitive medial preoptic nucleus (POM) of quail can be identified by the presence of a dense network of vasotocinergic fibers. This innervation is sexually differentiated (present in males only) and testosterone sensitive. The origin of these fibers has never been formally identified although their steroid sensitivity suggests that they originate in parvocellular vasotocinergic neurons that are found in quail only in the medial part of the bed nucleus striae terminalis (BSTm) and in smaller numbers within the POM itself. We report here that following injections of a retrograde tracer into the POM of male quail, large populations of retrogradely labeled cells can be identified in the BSTm. The POM also receives afferent projections from magnocellular vasotocinergic nuclei, the supraoptic and paraventricular nuclei. Double labeling for vasotocin immunoreactivity of the retrogradely labeled sections failed however to clearly identify magnocellular vasotocin immunoreactive cells that were retrogradely labeled from POM. In contrast a substantial population of vasotocin-immunoreactive neurons in the BSTm contained tracer retrogradely transported from the POM. These data therefore demonstrate that a significant part of the vasotocinergic innervation of the quail POM originates in the medial part of the BST. An intrinsic innervation could however also contribute to this network. This interaction between BSTm and POM could play a key role in the control of male-typical sexual behavior and in its sex dimorphism in quail. PMID- 12084410 TI - Expression of the 40 kDa catecholamine regulated protein in the normal and injured rat retina. AB - Catecholamine regulated protein 40 (CRP40) has been shown to be expressed in the central nervous system (CNS) of several mammalian species where it may function in a similar manner to members of the heat shock protein (HSP) family. Immunohistochemical and immunoblotting techniques were utilized to investigate whether CRP40 is expressed in normal rat retinas. In addition, changes in CRP40 expression were studied following optic nerve transection. The immunohistochemical results showed that CRP40 is expressed in the normal rat retina. The protein was found to be highly expressed in the ganglion cell layer (GCL), the inner nuclear layer (INL) and the outer plexiform layer (OPL). In addition, a low level of CRP40 was found in the inner plexiform layer (IPL), and in the inner segment layer (ISL). No expression was found in the outer nuclear layer (ONL) of normal rat retina. The immunoblotting results show that CRP40 expression decreased in a time-dependent fashion after the optic nerve transection. This decrease indicates that the expression of CRP40 is dependent on the neuron's normal physiological state and that it plays an important function in physiological and pathological conditions in the retina. PMID- 12084411 TI - Vasoactive intestinal polypeptide-containing elements in the olfactory bulb of the hedgehog (Erinaceus europaeus). AB - The distribution of vasoactive intestinal polypeptide (VIP)-immunopositive elements was analyzed in the olfactory bulb (OB) of the Western European hedgehog (Erinaceus europaeus) under light and electron microscopy. The immunoreactivity appeared in an abundant population of periglomerular cells of the glomerular layer, in interneurons of the external plexiform layer, and in a restricted group of deep short-axon cells of the internal plexiform layer, the granule cell layer and the white matter. In the glomerular layer, VIP-containing periglomerular cells constituted a population of non-GABAergic neurons and did not receive synapses from olfactory axons. In the EPL, VIP-immunoreactivity appeared in a morphologically heterogeneous population of GABAergic interneurons, most of them identified as satellite cells and Van Gehuchten cells. These interneurons exerted an abundant and selective innervation of the somata, primary and secondary dendrites of the principal mitral and tufted cells, but did not contact granule cells. Perisomatic innervation of the principal cells followed two different patterns. The first included 'normal' basket-like arrangements of VIP-containing varicosities surrounding the somata of mitral and tufted cells. In the second, a set of satellite cells gave rise to short dendritic shafts that embraced the somata of principal cells in an 'exuberant' basket-like arrangement. These two morphological patterns of perisomatic innervation of principal cells were correlated with a neurochemical specificity of the target. In this sense, the 'exuberant' basket-like structures were always found surrounding a subpopulation of principal cells that did not contain the calcium-binding protein parvalbumin (PV). By contrast, they were never found surrounding the subpopulation of PV containing principal cells, which only showed 'normal' basket-like structures. This study provides new data on the connectivity and neurochemical features of the hedgehog olfactory bulb and suggests that the olfactory circuits in this species are more complex than those described in other mammals. PMID- 12084412 TI - Localization of 5-HT2A, 5-HT3, 5-HT5A and 5-HT7 receptor-like immunoreactivity in the rat cerebellum. AB - Although serotonin (5-hydroxytryptamine, 5-HT) is known to exert a modulatory action on cerebellar function, our current knowledge of the nature of receptor subtypes mediating serotonergic activity in this part of the brain remains fragmentary. In this study, we report the presence and distribution of 5-HT3, 5 HT5A and 5-HT7 receptor-like immunoreactivity in the rat cerebellum using immunofluorescence histochemistry. 5-HT3 immunoreactivity was found in fibers sparsely distributed throughout the cerebellum. Most of them were seen in the cerebellar cortex as fine varicose 5-HT3-positive axonal processes. 5-HT5A immunoreactivity, on the other hand, was observed in neuronal somata of the cerebellar cortex and deep cerebellar nuclei. Based upon cell morphology and the use of cell-specific markers, Purkinje cells, molecular layer interneurons and Golgi cells were found to be 5-HT5A immunopositive. In addition, the use of cell specific markers allowed us to identify previously reported large 5-HT2A-positive cells in the granular layer as being Golgi cells. Finally, 5-HT7 immunoreactivity was observed only in Purkinje cells. Corroborating previous radioligand-binding, in situ hybridization and immunohistochemical studies, our data relate serotonin receptor subtypes to specific cerebellar cell types and may consequently contribute to the elucidation of serotonergic actions in the cerebellum. PMID- 12084413 TI - Newer atypical antipsychotic medication in comparison to clozapine: a systematic review of randomized trials. AB - The aim of this study was to evaluate the effectiveness of newer atypical antipsychotic drugs in comparison to clozapine for schizophrenia. Publications in all languages were searched from all relevant databases and all randomized controlled trials comparing clozapine with newer atypical drugs were included. The review and meta-analysis includes eight studies, most of them short in duration. Newer atypical drugs were broadly similar to clozapine when improvement was measured using a psychosis symptom rating scale or a global index. There was a trend for clozapine to be more effective than the others for positive symptoms, and less effective for the negative symptoms. The adverse effect profile of clozapine and newer atypical drugs was dissimilar: while clozapine produced more fatigue, hypersalivation, and orthostatic dizziness, new atypical drugs, with the exception of olanzapine, produced more extrapyramidal symptoms. As these results were obtained from few studies and a relatively small amount of patients, the equal effectiveness and tolerability of new atypical drugs in comparison with clozapine is not yet demonstrated. More trials of sufficient power, with longer duration, and measuring clinically important outcomes are urgently needed. PMID- 12084415 TI - D-Cycloserine added to risperidone in patients with primary negative symptoms of schizophrenia. AB - BACKGROUND: D-Cycloserine, a partial agonist at the glycine recognition site of the NMDA receptor has previously been shown to improve negative symptoms when added to conventional antipsychotics and to worsen negative symptoms when added to clozapine. The purpose of this study was to examine the effects of D cycloserine when added to risperidone on negative symptoms of schizophrenia. METHOD: Ten patients with schizophrenia who were treated with risperidone completed consecutive two week trials of placebo and four doses of D-cycloserine. Clinical assessments were videotaped and were scored by a rater who was blind to temporal sequence. RESULTS: D-Cycloserine at a dose of 50mg/day was associated with significant reduction in negative symptoms (mean=10%). Ratings of depression, extrapyramidal side effects, and cognitive function were unchanged. Serum concentrations of glutamate and serine increased significantly on this dose of D-cycloserine. CONCLUSIONS: This preliminary study suggests that combination of D-cycloserine, 50mg/day, with risperidone may improve negative symptoms of schizophrenia over a narrow dose range. The degree of improvement appears to be intermediate between improvement of negative symptoms observed with combination of D-cycloserine with conventional antipsychotics and worsening of negative symptoms observed with combination of D-cycloserine with clozapine in previous trials of identical design. PMID- 12084414 TI - Prolactin responses to acute clomipramine and haloperidol of male schizophrenic patients in a drug-free state and after treatment with clozapine or with olanzapine. AB - Atypical neuroleptics share a common feature, showing higher affinity for 5-HT2 receptors than for D2 dopamine receptors, but show considerable differences in their clinical and pharmacological properties. In clinical doses, they occupy serotonergic receptors near saturation, but show considerable differences regarding the D2 receptor occupancies, with clozapine showing the lowest degree of occupation. We assessed serotonergic and dopaminergic receptor responsiveness in two groups of male schizophrenic patients, one treated with the atypical neuroleptic clozapine (14 patients, doses 200-600 mg/d) and the other treated with olanzapine (11 patients, doses 10-30 mg/d). We measured the prolactin responses to the acute administration of a serotonergic drug, clomipramine, and a dopaminergic one, haloperidol. Tests were first performed in the drug-free state, and were repeated after the patients had been treated with stable doses of either drug for six weeks. Clomipramine administration induced significant increases of prolactin in the drug-free state. These responses were eliminated after treatment of the patients with either drug, thereby indicating a high 5-HT receptor occupancy by both clozapine and olanzapine. The prolactin responses to haloperidol were not altered after treatment with clozapine, but were significantly reduced after the olanzapine treatment. The baseline prolactin levels were not influenced by clozapine treatment, and were moderately but significantly increased after treatment with olanzapine. The results indicate that there is a difference between the two drugs in their capacity to block dopamine receptors at the hypothalamus-pituitary level, and match the results obtained by SPECT receptor binding studies for striatal dopamine receptors. PMID- 12084416 TI - Sexual side effects of novel antipsychotic medications. AB - BACKGROUND: The novel antipsychotic medications offer a more favorable extrapyramidal side effect profile than conventional agents. It is uncertain that the novel antipsychotics have a benefit in terms of sexual side effects. METHODS: We prospectively administered a survey of sexual functioning to 25 male patients with DSM-IV schizophrenia, taking conventional and novel antipsychotics. Contrasts were made between three treatment groups: clozapine (CLOZ), risperidone (RIS), and a combined haloperidol/fluphenazine (HAL/FLU) group. RESULTS: A decrease in overall sexual functioning was reported in all medication groups (40 71%). The majority of subjects taking RIS or HAL/FLU reported a decline in one or more aspects of sexual functioning. Examining specific aspects of sexual functioning revealed that, a decline in sexual interest was significantly less common on CLOZ compared to RIS (0 vs. 64%; chi(2)=6.1, df=1, p=0.01) or HAL/FLU (0 vs. 67%; chi(2)=5.2, df=1, p=0.02), while a decline in the erectile frequency was significantly more common on RIS compared to CLOZ (40 vs. 93%; chi(2)=6.2, df=1, p=0.01) or HAL/FLU (50 vs. 93%; chi(2)=4.8, df=1, p=0.03) (0%). For enjoyment of orgasm and ejaculatory volume, significantly fewer CLOZ compared to RIS subjects reported a decline (20 vs. 86%; chi(2)=7.4, df=1, p=0.01). CONCLUSIONS: Sexual side effects are common clinically pertinent adverse effects associated with both novel and conventional antipsychotic medications. They deserve increased attention in clinical work and future research with emerging antipsychotic drugs. PMID- 12084417 TI - Typical antipsychotic drugs -- D(2) receptor occupancy and depressive symptoms in schizophrenia. AB - We tested the hypothesis that the degree of striatal dopamine D(2) receptor blockade induced by typical antipsychotic treatment directly correlates with the presence and severity of depressive symptoms in schizophrenia. Clinical and [(123)I]-IBZM single-photon emission tomography (SPET) scan data obtained from 18 typical antipsychotic treated schizophrenic patients was analysed to evaluate the relationship between striatal D(2) receptor occupancy and the depressive subscale of the Brief Psychiatric Rating Scale (BPRS-D). Striatal D(2) receptor occupancy by typical antipsychotic drugs was significantly positively correlated with BPRS D scores (r=0.52, p=0.025). This study suggests that high striatal dopamine D(2) blockade by typical antipsychotic drugs may contribute to the emergence of depressive symptoms in typical antipsychotic treated schizophrenic patients. PMID- 12084418 TI - Personal evaluation of transitions in treatment (PETiT):a scale to measure subjective aspects of antipsychotic drug therapy in schizophrenia. AB - The paper describes the development and preliminary testing of a scale designed to capture aspects of subjective responses to, and tolerability of antipsychotic drugs, treatment adherence, and impact of antipsychotic drug therapy on the quality of life of individuals treated for schizophrenia. Using empirical study approaches and qualitative methods of data analysis, twelve themes were initially identified as relevant to the quality of life of individuals during antipsychotic drug therapy. Based on these dimensions, in the second phase, a 30 item self report questionnaire was constructed and field tested in a community based, heterogeneous sample of schizophrenic patients (n=335). The scale was easy to self-administer (in 2-5 min) and perceived as user-friendly by patients. Correlational analysis revealed a high internal consistency (Cronbach's alpha=0.92) and split half reliability (Spearman-Brown coefficient of 0.85). The scale scores were able to distinguish subjects with lower and higher rates of treatment adherence, and factor analysis confirmed the robustness of the original construct. Repeated administration of the scale in a sub-sample of clinically stable schizophrenic population (n=71) revealed a test-retest reliability coefficient of 0.97 (P<0.001); and repeat administration at quarterly intervals in a sample of patients receiving active treatment (n=54) indicated a significant increases in mean scores (t=6.2, df=53, P<0.005), suggesting that the scale was sensitive to changes in patients' clinical status. Based on these preliminary data, PETiT could be considered as a potentially useful tool for measuring client centred outcomes in clinical practice, drug trials, quality assurance programs and interventional research involving schizophrenic patients. PMID- 12084419 TI - Initiation of daily smoking and nicotine dependence in schizophrenia and mood disorders. AB - This study replicates, using more refined methodology, the indications of prior studies that patients with schizophrenia show a greater frequency of tobacco smoking than patients with mood disorders. The sample included 66 patients with schizophrenia and 51 patients with a mood disorder who were admitted at a state hospital in Kentucky. The control group included 404 community subjects. Ever daily smoking was studied using logistic regression. Survival analyses of age of onset of daily smoking (AODS) were performed controlling for several variables including education level. Nicotine dependence was measured with a scale. The prevalence of ever and current daily smoking was respectively 92 and 83% for patients with schizophrenia, 78 and 65% for patients with mood disorders, and 47 and 26% for controls. Before the age of 20, the three populations appear to have a similar risk of smoking initiation. However, after the age of 20, the initiation rate of daily smoking for patients with schizophrenia was higher than in patients with a mood disorder, or controls. Among daily smokers, there were no differences in nicotine dependence between patients with schizophrenia and those with a mood disorder. Schizophrenia was associated with a greater probability of ever daily smoking than mood disorders and with higher rates of initiation of daily smoking after 20 years old. PMID- 12084420 TI - Schizophrenia and tobacco smoking: a replication study in another US psychiatric hospital. AB - A prior study in a US state hospital suggested that schizophrenia is more closely associated with tobacco smoking when compared with other severe mental illnesses. This second study, in a neighborhood hospital, tries to (1) replicate that schizophrenia is associated with smoking and heavy smoking, and (2) rule out that this relationship is explained by substance use. The methodology was very similar to the first study. The sample included 588 inpatients. Logistic regression was used to develop models of variables associated with smoking or heavy smoking. The frequency of current smoking for the total, schizophrenic and non-schizophrenic samples were respectively 71, 75, and 55%. The sequence of frequencies from the highest to lowest was the same as in the first study: male schizophrenic patients, male non-schizophrenic patients, female schizophrenic patients and female non-schizophrenic patients. This second study consistently replicated the relationship between schizophrenia and smoking (after correcting for other variables) including history of alcohol and drug abuse or dependence. Only one of two definitions of heavy smoking showed a significant association with schizophrenia. In summary, these two studies suggest that schizophrenia is strongly associated with smoking. Neither substance use, antipsychotics, nor institutionalism can explain this relationship. PMID- 12084421 TI - Smoking in schizophrenia -- all is not biological. AB - High rate of tobacco smoking reported in schizophrenia has been related to the effect of nicotine on the neurobiology of schizophrenia. Nicotine is said to alleviate psychotic symptoms in some patients. The relationship between smoking and psychiatric status may not be simply a biological one as several sociocultural and economic factors could influence smoking behaviour. In this study in India on 286 urban male outpatients with schizophrenia, only 38% were found to be current smokers. This was significantly more than in other psychiatric patients studied (major affective disorders and non-psychotic disorders) but not medically ill controls and not higher than the rates for the general male population in India. Smokeless use of tobacco was infrequent in the study population. More than half of the patients did not experience any positive effects due to smoking. Lack of economic independence and restrictions imposed by the family appeared to be crucial factors that controlled the prevalence of smoking among schizophrenia patients. As smoking is a leading cause of preventable morbidity and mortality, there is a serious need to review the neurobiological issue of smoking in schizophrenia considering the influence culture and social practices could have upon the behaviour. PMID- 12084422 TI - Serotonergic function and suicidal behavior in schizophrenia. AB - Recent studies suggest that altered serotonergic (5-HT) function, as assessed by lower prolactin (PRL) response to fenfluramine (FEN), a specific 5-HT releaser and uptake inhibitor, is associated with suicidal behavior in either depressed and personality disordered patients. The purpose of this study was to investigate, in schizophrenic patients, the relationship between suicidal behavior and PRL response to D-fenfluramine (D-FEN). A D-FEN test was performed in 18 healthy controls and 33 drug-free DSM-IV schizophrenic patients (12 with a history of suicide attempts, 21 without it). Schizophrenic patients with a history of suicide attempts showed a lower PRL response to D-FEN (Delta PRL) compared to schizophrenic patients without such history (P<0.04) and also compared to healthy controls (P<0.0003). Delta PRL did not differentiate schizophrenic patients without suicide attempts and controls. These findings could not be explained by PRL basal hormonal levels, age, sex, menstrual status, demographic or clinical characteristics. These results suggest that PRL response to D-FEN is a marker of suicidal tendencies also in schizophrenia, supporting the hypothesis that a dysfunction in serotonergic function is associated with suicidal behavior regardless of the psychiatric diagnosis. PMID- 12084423 TI - The Kraepelinian dichotomy: preliminary results of a 15-year follow-up study on functional psychoses: focus on negative symptoms. AB - In a 15-year follow-up study, we used a comparative approach to assess course and outcome for all functional psychoses. The presented results focus on negative symptoms and refer to a sample of 76 patients with schizophrenia, 38 patients with a schizoaffective disorder and 32 patients with an affective disorder according to ICD-9. These patients were assessed at their first psychiatric hospitalization and 15 years later. In summary, the findings indicate that the course and outcome of schizophrenia is less favorable than that of affective and schizoaffective disorders. Negative symptoms occurred in all functional psychoses, but were more frequent and prominent in the schizophrenic group than in the other two diagnostic groups at any time of assessment. Narrower concepts of negative symptoms, conceptualized as the deficit syndrome, seem to be specific for schizophrenia and appear quite rarely in patients with affective psychoses. Overall, our study supports Kraepelin's original hypothesis that bifurcated the psychoses into the affective psychoses and schizophrenia, whereby the latter have a more deleterious long-term course and outcome. PMID- 12084424 TI - The relationship between positive symptoms and instrumental work functioning in schizophrenia: a 10 year follow-up study. AB - This longitudinal investigation was designed to determine the association between the positive symptoms of psychosis and instrumental work functioning among patients with schizophrenia in comparison to patients with affective disorders. 173 participants were assessed prospectively for the presence of psychosis and concurrent work adjustment using a series of standardized measures at four consecutive follow-ups over a 10 yr period. The data demonstrate a significant relationship between psychosis and increased impairment in work functioning across diagnostic groups for three of the four follow-up periods (p<0.01), with this relationship being most consistent for the schizophrenia patients throughout the course of their disorder (p<0.01). The more severely psychotic patients, regardless of diagnosis, are least likely to be working effectively; however, the debilitating effect of psychosis on work adjustment is most evident for schizophrenia patients. PMID- 12084425 TI - Symptom patterns and subgrouping of schizophrenic patients: significance of negative symptoms assessed on admission. AB - This study subgroups schizophrenic patients based on symptoms assessed on admission and examines the validity of the subgrouping using follow-up data and other clinical outcome variables. Schizophrenic patients (n=163) from consecutive admission received ratings on the positive and negative syndrome scale (PANSS) on admission and during a 1-year follow-up course. An exploratory graphic analysis on the admission PANSS derived four symptom dimensions: negative symptoms, disorganized thought, hostility/excitement and delusions/hallucinations. This yielded two subgroups of patients on admission, a group with marked negative (GWNEG) and a group without marked negative (GONEG) symptoms. Compared with the GONEG, the GWNEG had a poorer recovery rate, more impairment in attention, a slower response of the delusion/hallucination symptoms to neuroleptic treatment and a longer duration of index hospitalization. At a one-year follow-up, the GWNEG assessed on admission had persistently higher scores on the negative symptom and disorganized thought syndromes, less relapse rate, a shorter duration on job, as well as worse social functioning than the GONEG. Thus, the GONEG might comprise patients having a pure paranoid syndrome with quick and better treatment response, while the GWNEG comprises patients with the blunt-disorganization syndrome having a poorer outcome. PMID- 12084426 TI - Validity of the depressive dimension extracted from principal component analysis of the PANSS in drug-free patients with schizophrenia. AB - Depressive symptoms frequently occur during the course of schizophrenia. This study explored the relationships between the schizophrenia symptomatology and three measures of depression. Eighty-one drug-free inpatients with acute schizophrenia were assessed with the positive and negative syndrome scale (PANSS), the Calgary depression scale for schizophrenia (CDSS), and the Hamilton rating scale for depression (HAM-D). The depressive subscale of PANSS (PANSS-D) was also considered as a third scale for measuring depression. A principal component analysis (PCA) of PANSS items identified five clinical dimensions of schizophrenia called 'negative', 'positive', 'anxio-depressive', 'excitement', and 'disorganisation and others'. Our anxio-depressive dimension (PANSS-ad) was strictly identical with the PANSS-D. Scores on CDSS and HAM-D were highly inter correlated and highly correlated with the PANSS-ad. Furthermore, while scores on CDSS were correlated only with this dimension, scores at HAM-D were also positively correlated with the negative dimension and negatively correlated with the excitement dimension. In conclusion, our results suggest that PANSS evaluation itself may be sufficient to give a correct approximation of the depression in patients with schizophrenia. However, depression scales are of course needed to assess specifically depressive symptoms in patients with schizophrenia; hence, the CDSS could be a more specific instrument than HAM-D. PMID- 12084427 TI - PANSS factors and scores in schizophrenic and bipolar disorders during an index acute episode: a further analysis of the cognitive component. AB - To investigate the factor structure of psychotic symptoms, we compared the clinical characteristics of manic patients with those of schizophrenic patients evaluated with positive and negative syndrome scale (PANSS). The clinical symptoms of 148 bipolar patients and 86 schizophrenic patients hospitalized for an index psychotic episode were assessed. Schizophrenic patients showed more positive and cognitive symptoms than bipolars. The factor analysis of the two PANSS scores showed a three-factor solution with 'positive', 'negative' and 'mixed' depressive-activated factors for bipolars and 'positive', 'negative' and 'depressive' factors for schizophrenics. In both groups, the 'cognitive cluster' loaded on the first 'positive' factor while the 'lack of insight' (LOI) has a different meaning in the two groups, more related to the positive symptoms in the bipolar patients and more related to the negative symptoms in the schizophrenic patients. This finding suggests that LOI could be a non-unitary phenomenon in psychoses and it should be further explored to better elucidate differences in symptom structures between schizophrenic and bipolar disorders. PMID- 12084428 TI - The dimensional symptom structure of schizophrenia and its association with temperament and character. AB - Recent studies suggest that personality may influence symptom expression and social functioning in schizophrenia. This study investigated the relationships between personality and symptom dimensions in schizophrenia patients. Fifty-two schizophrenia patients and 25 five healthy subjects were assessed using the Temperament and Character Inventory (TCI). The patients were also assessed for positive and negative symptoms using SAPS and SANS and scored according to Andreasen's (1995: Andreasen, N.C., Arndt, S., Alliger, R., Miller, D., Flaum, M. 1995. Symptoms of schizophrenia. Methods, meanings, and mechanisms. Arch. Gen. Psychiatry, 52, 341-351) classical three dimensional model and by the five dimensional model of Toomey et al. (1997: Toomey, R., Kremen, W.S., Simpson J.C., Samson, J.A., Seidman, L.J., Lyons, M.J., Faraone, S.V., Tsuang, M.T. 1997. Revisiting the factor structure for positive and negative symptoms: evidence from a large heterogeneous group of psychiatric patients. Am. J. Psychiatry, 154, 371 377). Comparisons between patients and controls revealed significant differences on various TCI scores consistent with a global disorganization of personality in schizophrenia involving both basic neurophysiological and potentially genetically determined traits (i.e. temperament) and developmental aspects of personality (i.e. character). Correlation analysis showed distinct associations between symptoms and personality dimensions. The results suggest that the negative and disorganized dimensions of schizophrenia are related temperamental factors, whereas the psychotic symptoms are more related to characterological abnormalities. The observed patterns of associations also underline the heterogeneity of the classical negative and positive dimensions of schizophrenia. PMID- 12084429 TI - Social and clinical consequences of cognitive deficits in early psychosis: a two year follow-up study of first-admitted patients. AB - OBJECTIVE: To explore whether baseline memory and executive deficits predicted poor social and clinical outcome over the 2 years following a first admission for psychosis, regardless of categorical diagnosis. METHOD: Cognitive functioning was assessed in first-admitted subjects with psychosis (n=35) with a neuropsychological battery of tests measuring executive, language and memory functions. Social and clinical outcome were assessed at 6-monthly intervals over a two-year follow-up using multiple sources of information. RESULTS: A dose response relationship was found between visual and verbal memory performance at first admission and clinical outcome over the 2-year follow-up: the poorer the memory performance, the more likely the risk of presenting with psychotic symptoms and rehospitalization. Poor baseline performance on the WCST executive function predicted better medication adherence. No association was found between cognitive performance and occupational or residential outcome. CONCLUSION: Cognitive performance was a better predictor of clinical than social outcome in this sample of first-episode patients. The association between cognitive deficits and poor social outcome may be more marked in subjects with chronic psychosis than in first-episode subjects. The finding that cognitive deficits predict better medication adherence is in need of further exploration. PMID- 12084430 TI - Independence and overlap among neurocognitive correlates of community functioning in schizophrenia. AB - Existing literature on the neurocognitive correlates of community functioning in schizophrenia has not adequately focused on the relationships among the correlated variables. In a sample of 40 outpatients with schizophrenia and related disorders, we studied two sets of variables that we expected to be related to broad ratings of community functioning: (1) the Wechsler Adult Intelligence Scale-III (WAIS-III) index scores for verbal comprehension, perceptual organization, working memory, and processing speed; and (2) positive, negative, disorganized and affective symptom variables. Of the WAIS-III index scores, working memory and processing speed entered a stepwise regression, together accounting for substantial variance in functional ratings (R(2)=0.37). However, only processing speed remained significantly associated with community functioning after controlling for the other indexes. In relation to community functioning, the remaining indexes appeared to be overlapping markers of general cognitive ability, rather than specific measures of discrete cognitive domains. Addition of positive and negative symptom variables in a further analysis greatly increased the explained functional variance (R(2)=0.65). Processing speed overlapped substantially with negative symptoms in predicting functioning, while the other WAIS-III indexes were independent of symptomatology. Results illustrate the importance of (1) knowing which neurocognitive variables have specific relationships to community functioning and which reflect the influence of more general cognitive ability in daily life, and (2) appreciating areas of overlap and independence among classes of correlates of functioning, such as neurocognitive and symptom variables. This improved understanding has implications for predictive models of community functioning, for cognitive rehabilitation and deficit compensation strategies, and for assessment practice. PMID- 12084431 TI - Positive and negative schizotypy in a student sample: neurocognitive and clinical correlates. AB - Positive and negative schizotypy may represent discrete factors or dimensions. To determine if distinct neurocognitive profiles are associated with these dimensions or factors, we classified university students on the basis of positive and negative schizotypal symptoms and conducted separate analyses. Following prior work in the neuropsychiatric literature, we predicted that subtle prefrontal deficits would be selectively associated with negative schizotypal personality features in a nonclinical student sample. We also investigated the relationship between positive/negative schizotypy and associated clinical states or personality dimensions including antisocial personality disorder, obsessive compulsive personality traits, generalized and social anxiety, empathy, and impulsivity. Classification of subjects into positive and negative schizotypy groups revealed distinct neurocognitive and clinical profiles. We observed a positive relation between measures of temporolimbic dysfunction, impulsivity, antisocial behavior, and positive schizotypal phenomena. Negative schizotypy was associated with subtle performance deficits on measures of frontal executive function, increased social anxiety, and obsessive-compulsive phenomena. Findings are consistent with the contention that positive and negative schizotypy represent discrete factors. PMID- 12084432 TI - Extreme weight gain in a youth with schizophrenia: risk/benefit considerations. PMID- 12084433 TI - The characteristics of clozapine-induced fever. PMID- 12084434 TI - Olanzapine-associated type 2 diabetes mellitus. PMID- 12084435 TI - Depression measures and motor side-effects in patients with acute schizophrenia. PMID- 12084436 TI - Primary progressive multiple sclerosis: increasing clarity but many unanswered questions. AB - Heterogeneity in the clinical course of multiple sclerosis (MS) is well recognised and patients following a primary progressive course, 10-15% of the MS population, have a distinct clinical and paraclinical phenotype. This review examines recent advances in our understanding of this subgroup of patients and examines the new criteria to be applied in diagnosis. It also highlights developments in genetic, immunological, magnetic resonance and pathological aspects of the disease, whilst also outlining the results of recent therapeutic trials. PMID- 12084437 TI - Caspase inhibition by Z-VAD increases the survival of grafted bone marrow cells and improves functional outcome after MCAo in rats. AB - Marrow stromal cells (MSCs) transplantation into brain has been employed to treat experimental ischemia. However, MSCs undergo apoptosis and few survive in the ischemic brain. We test the hypotheses that coadministration of bone marrow cells (BMCs) with a cell-permeable inhibitor of caspases, Z-Val-Ala-DL-Asp fluoromethylketone (Z-VAD), into the ischemic boundary zone (IBZ) of brain promotes BMCs survival and improve outcome. Experimental groups consist of: 24 h after MCAo, either phosphate-buffered saline (PBS, n=4), dead BMC (n=4), fresh BMC (n=10), Z-VAD only (n=4), or BMC with Z-VAD (n=6) were intracerebrally injected. BMCs were harvested from donor adult rats labeled with bromodeoxyuridine (BrdU). Rats were subjected to an adhesive-removal somatosensory and motor-rotarod functional tests before MCAo and at 1 and 7 days after MCAo. Rats treated with a combination of Z-VAD and BMCs exhibited significant improvement in the adhesive-removal test at 7 days compared with the control group (combined MCAo+PBS and MCAo+dead BMC) (p<0.01), and the numbers of BrdU-BMC increased (p<0.05) and apoptotic cells decreased (p<0.05) compared with BMC alone transplantation. Our data suggest that intracerebral coadministration of BMC with Z-VAD enhances the survival of grafted BMC and improves neurological functional recovery after MCAo. PMID- 12084438 TI - Association between genetic polymorphism of angiotensin-converting enzyme gene and Parkinson's disease. AB - This study was designed to investigate the hypothesis that deletion/insertion (D/I) polymorphism of the angiotensin-converting enzyme (ACE) gene may contribute to increased risk of Parkinson's disease (PD). A case-control study was carried out to examine the association between the ACE genotype and the allele frequency in 127 sporadic PD patients compared with 198 healthy controls. The frequency of the homozygote DD genotype of the ACE gene was significantly increased in patients with PD than in the controls (chi(2)=6.09, p=0.048), despite that there was no significant difference in D/I allele frequency (chi(2)=2.25, p=0.133). Moreover, PD patients carrying the homozygote DD genotype were 1.13 times more frequent than subjects without the DD genotype (chi(2)=5.67, 95% CI=1.01-1.25, p=0.017). A stepwise logistic regression analysis of the presence of the DD genotype and data on risk factors for PD confirmed that the homozygote DD genotype was a modest independent risk factor for PD (OR=1.32, 95% CI=1.12-2.16). In addition, there was a trend of increasing number of DD genotype in older PD patients and the modest risk factor of DD genotype in PD was due to the significant difference of the DD homozygosity in old patients with onset age at or after 60 years. In conclusion, results of our study support the hypothesis that the ACE gene may indicate genetic susceptibility to PD, particularly in older individuals. PMID- 12084439 TI - Abnormalities of rate-corrected QT intervals in Parkinson's disease-a comparison with multiple system atrophy and progressive supranuclear palsy. AB - A number of patients with Parkinson's disease (PD) and multiple system atrophy (MSA), in whom sudden death does occur occasionally, have QT or rate-corrected QT (QTc) interval prolongation on electrocardiogram (ECG). Although these QT or QTc interval abnormalities are likely related to autonomic dysfunction, the pathophysiology remains unknown. The aim of this study was to compare the degree of QTc interval prolongation among akinetic-rigid syndromes, namely PD and related disorders, and to evaluate the relationship between QTc prolongation and severity of autonomic dysfunction. Thirty-four patients with PD, 22 with MSA, 11 with progressive supranuclear palsy (PSP) and 30 healthy controls underwent standard autonomic function tests, and electrocardiography variables (RR, QT and QTc intervals) were measured by an ECG recorder with an automated analyzer. The relationship between QTc interval and cardiovascular reflex tests were also analyzed. Orthostatic hypotension and decreased heart rate in response to respiratory stimuli were prominent in MSA, while these were relatively mild in PD. Unlike the RR and QT intervals, the QTc interval significantly differed among all groups (p<0.01). The QTc interval was significantly prolonged in PD (409+/-17 ms; p<0.001) and MSA (404+/-14 ms; p<0.05) compared with healthy controls (394+/ 19 ms). Neither autonomic dysfunction nor QTc interval prolongation was evident in PSP. QTc intervals and cardiovascular reflexes did not correlate, except for Valsalva ratio. The QTc interval was obviously prolonged in PD patients to an extent that could not be accounted for simply by autonomic dysfunction levels. MSA patients showed slightly prolonged QTc intervals in spite of marked cardiovascular autonomic dysfunction. Abnormalities of the QTc may reflect the degeneration of cardioselective sympathetic and parasympathetic neurons that cannot be fully captured by cardiovascular autonomic function tests. PMID- 12084440 TI - Signaling pathway for radiation-induced apoptosis in the lymphoblasts from neuronopathic (type A) and non-neuronopathic (type B) forms of Niemann-Pick disease. AB - Deficient activity of human lysosomal hydrolase, acid sphingomyelinase (ASM), results in the neuronopathic (type A) and non-neuronopathic (type B) forms of Niemann-Pick disease (NPD). A deficiency of ASM is known to deprive lymphoblasts of their response to apoptotic induction by X-ray irradiation. To elucidate the genetic heterogeneity of apoptotic induction in NPD cells, we investigated radiation-induced apoptosis of lymphoblasts in patients with type A (genotype: IVS3-2A-G/IVS3-2A-G) and type B (genotype: S436R/S436R) NPD. Epstein-Barr virus (EBV)-transformed lymphoblasts established from a patient with type A NPD, a patient with type B NPD and a normal control were irradiated with 20 Gy and incubated for 24 h. The cells were harvested and the morphological features of apoptosis were observed with DNA-specific fluorochrome bis-benzimide. Exposure of lymphoblasts to 20 Gy of radiation resulted in 25-30% apoptosis of total cells in normal lymphoblasts, 8-9% apoptosis in type A NPD and 20-27% apoptosis in type B NPD. The radiation-induced apoptotic response in the lymphoblasts of type A NPD was significantly different from that of the normal lymphoblasts (P<0.0005). On the other hand, the radiation-induced apoptotic response in type B NPD was not markedly different from that in normal lymphoblasts (P=0.624). In the patient with type B NPD, the signaling pathway for radiation-induced apoptosis was preserved in lymphoblasts, which suggests that the extent of cell signaling system disturbance due to ASM deficiency may be related to the phenotypes in types A and B NPD. PMID- 12084441 TI - Muscle MRI findings of HTLV-1-associated myelopathy. AB - We report here the muscle MRI findings in two patients with human T-cell lymphotropic virus type I-associated myelopathy (HAM). It is known that thigh muscles are vulnerable in HAM patients, but detailed information about the affected muscles has not been available. Muscle MRI findings of these patients showed that thigh muscles, especially adductor magnus, and semimembranosus muscles were severely affected, but lower leg muscles were comparatively preserved. In these affected muscles, neurogenic changes were observed by EMG. We concluded that muscle MRI is very useful to estimate the affected muscles in HAM patients. PMID- 12084442 TI - Sources and reasons for delays in the care of acute stroke patients. AB - OBJECTIVE: This study aimed to identify sources and reasons for delays in the care of our acute stroke patients. METHODS: Data on time interval from symptom onset or awareness to initial presentation, to neurology assessment, to performance of cranial CT scan, and demographic and medical factors associated with delays among stroke patients admitted at St. Luke's Medical Center from May to October 2000 were obtained by interview and record review. RESULTS: Of 259 patients (mean age 61.5+/-13.6 years, 43% females), 63% had infarction (INF), 32% intracerebral hemorrhage (ICH) and 5% subarachnoid hemorrhage (SAH). Fifty-nine percent presented within 3 h of symptom onset or awareness, 73% within 6 h (median=2 h). Patients with ICH presented earlier than those with infarction. Reasons for delayed consultation included failure to recognize symptoms as serious and stroke-related. A non-neurologist was initially consulted in 97% of cases. Median delay from presentation to neurology evaluation was 7.5 h. Median time from presentation to brain imaging was significantly shorter for patients brought to CT-equipped facilities (2 h) than for those needing transfer to other hospitals (11.5 h). CONCLUSIONS AND RECOMMENDATIONS: Patient delay in presentation is only one cause of delay in acute stroke care. Longer delays arise from healthcare-related factors such as delays in neurologist referral and neuroradiologic diagnosis. Professional and public education on the necessity of early neurologic evaluation and patient transport to CT-equipped "Stroke Centers" is recommended. PMID- 12084443 TI - Peripheral silent periods in essential tremor. AB - BACKGROUND: Pathophysiology of essential tremor (ET) is controversial. In the present study, peripherally induced silent period (SP) in ET patients is studied. AIMS AND OBJECTIVES: To study if the peripherally induced SP was different in ET patients as compared to age matched healthy controls. MATERIAL AND METHODS: 24 patients of ET diagnosed according to diagnostic criteria of Louis et al. [Neurology 50 (1998) 1351] (mean age 45.37+/-14.86 years) and an equal number of healthy controls (mean age 36.21+/-15.72 years) were recruited for the study. Peripherally induced SP was recorded according to the methods already described. Student's t-test and Wilcoxon sign rank test were used for statistical analysis. RESULTS: The peripheral SP was 50.29+/-50.15 and 93.04+/-35.93 ms (p=0.0014) in ET patients and controls respectively. CONCLUSION: Our study shows that peripheral silent period is shorter in patients of ET as compared to normal individuals. PMID- 12084444 TI - Hyperintense and hypointense MRI signals of the precentral gyrus and corticospinal tract in ALS: a follow-up examination including FLAIR images. AB - In amyotrophic lateral sclerosis (ALS) patients, hyperintense signals at the subcortical precentral gyrus in brain fluid attenuated inversion recovery (FLAIR) MR images have been found more frequently than in controls. Quantitative analysis has revealed a significant increase of the FLAIR-magnetic resonance imaging (MRI) signal at the subcortical precentral gyrus of ALS patients compared to healthy controls. In addition, hypointense signals at the rim of the precentral gyrus in FLAIR and T2-weighted images have been shown in ALS patients. In 17 ALS patients, we evaluated hyperintense signals in T2-, T1-, proton density-weighted and FLAIR MR images, and hypointense signals in T2-weighted and FLAIR images 15.7+/-3.0 months after the initial examination by visual scoring. In FLAIR images, a quantitative analysis was added. The visual scores of hyperintense signals along the corticospinal tract did not change significantly in all sequences. However, the quantitative evaluation of FLAIR images revealed a significant increase of the signal intensity at the subcortical precentral gyrus (p<0.005). In addition, the frequency of the visually evaluated hypointense signals at the precentral gyrus increased significantly (p<0.05). The change of MR results did not correlate with the change of clinical parameters. In ALS patients, the increase of the quantified MRI signal at the subcortical precentral gyrus in FLAIR images and the increase of hypointense signals at the rim of the precentral gyrus corroborate the hypothesis that these signals are related to the upper motor neuron degeneration in ALS. Their specificity and clinical relevance have to be clarified further. PMID- 12084445 TI - Voxel-based comparison of regional cerebral glucose metabolism between PSP and corticobasal degeneration. AB - OBJECTIVES: Progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD) are neurodegenerative disorders that may be accompanied by dementia and parkinsonism as clinical symptoms. The purpose of this study was to elucidate cerebral metabolic differences of these two diseases with cognitive impairments by [18F] fluorodeoxyglucose (FDG) and positron emission tomography (PET). METHODS: A total of 12 patients with PSP (age: 62.8+/-6.0 years old, m: 7, f: 5, Mini-Mental State Examination (MMSE): 23.4+/-2.6), 12 patients with CBD (age: 64.8+/-6.3 years old, m: 6, f: 6, MMSE: 22.9+/-4.5), and age-matched healthy subjects (normal control (NC)) (age: 63.8+/-7.7 years old, m: 7, f: 5) were subjected to FDG-PET to obtain glucose metabolic images. We compared regional cerebral metabolic images by a voxel-by-voxel analysis with statistical parametric mapping (SPM) among PSP, CBD, and NC subjects, and evaluated differences of hypometabolic regions. RESULTS: The patients with PSP showed reduced cerebral glucose metabolism in the medial and lateral frontal gyri, basal ganglia, and midbrain compared with NC, whereas the patients with CBD showed significant reduction in the parietal lobes (p<0.001). SPM also revealed parietal hypometabolism in CBD patients compared with PSP patients (p<0.001). CONCLUSIONS: The predominant parietal glucose metabolic reduction in CBD patients was different from previously reported findings. This finding would be the characteristic substance of patients with CBD accompanying cognitive impairments. Our findings suggest that measurement of glucose metabolism by PET and a voxel based analysis is useful to understand the pathophysiology of these two diseases with cognitive impairments. PMID- 12084446 TI - Spectroscopic data following stroke reveal tissue abnormality beyond the region of T2-weighted hyperintensity. AB - Cerebral tissue with T2 magnetic resonance imaging (MRI) abnormalities following stroke is generally considered infarcted, while surrounding regions with normal MRI appearance are believed to be healthy. To assess whether these surrounding regions consist of normal tissue, we explored the distribution of N acetylaspartate (NAA) and lactate within and around the hyperintense area on T2 weighted MRI using proton MR spectroscopy. The study was carried out in 25 patients with middle cerebral artery occlusion imaged between 1 and 42 days after stroke onset. NAA/choline (Cho) ratios were significantly reduced in both areas of T2 hyperintensity and in surrounding tissue. The reduction was greater in the region of T2 hyperintensity than in the surrounding region (-50% vs. -28%, respectively) and was unrelated to the delay after the ictus. Lactate/Cho ratios increased massively within the abnormal T2 area, but did not differ from control values beyond the margin of hyperintensity. Overall data indicate that T2 visible lesions on MRI do not infer the entire injured tissue. PMID- 12084447 TI - Gender differences in cerebral glucose metabolism: a PET study. AB - OBJECTIVE: Some studies have examined gender differences in brain function based on cerebral blood flow and cerebral metabolism by using positron emission tomography (PET). However, the findings of these studies are controversial and most of them were analyzed by the regions of interest (ROIs) method. Here, we evaluated gender differences of cerebral glucose metabolism under the resting state in a voxel-based analysis. METHODS: We studied 44 healthy volunteers (22 females, 63.0+/-6.3 years, and 22 males, 63.1+/-8.4 years). Cerebral glucose metabolic images were obtained with (18)F-fluorodeoxyglucose (FDG) and PET. All individual data were transformed to standard brain space and the male and female groups were compared using statistical parametric mapping (SPM). RESULTS: The males had significantly higher glucose metabolism in the right insula, middle temporal gyrus, and medial frontal lobe than the females. Glucose metabolism in the hypothalamus was significantly higher in females than in males. There was a significant correlation between aging and glucose metabolism in the left thalamus in males and in the left caudate nucleus and hypothalamus in females. In males, but not females, there was a significant asymmetry between the bilateral hemispheres. CONCLUSION: We found that there were obvious gender differences in regional cerebral glucose metabolism and this is the first report of higher glucose metabolism in the hypothalamus in females than in males. PMID- 12084448 TI - Ifosfamide encephalopathy presenting with asterixis. AB - CNS toxic effects of the antineoplastic agent ifosfamide (IFX) are frequent and include a variety of neurological symptoms that can limit drug use. We report a case of a 51-year-old man who developed severe, disabling negative myoclonus of the upper and lower extremities after the infusion of ifosfamide for plasmacytoma. He was awake, revealed no changes of mental status and at rest there were no further motor symptoms. Cranial magnetic resonance imaging and extensive laboratory studies failed to reveal structural lesions of the brain and metabolic abnormalities. An electroencephalogram showed continuous, generalized irregular slowing with admixed periodic triphasic waves indicating symptomatic encephalopathy. The administration of ifosfamide was discontinued and within 12 h the asterixis resolved completely. In the patient described, the presence of asterixis during infusion of ifosfamide, normal laboratory findings and imaging studies and the resolution of symptoms following the discontinuation of the drug suggest that negative myoclonus is associated with the use of IFX. PMID- 12084449 TI - Hyperreflexia in axonal Guillain-Barre syndrome subsequent to Campylobacter jejuni enteritis. AB - We describe a patient with the acute motor axonal neuropathy (AMAN) form of Guillain-Barre syndrome (GBS), who showed generalized hyperreflexia. A 24-year old man developed acute paralysis following Campylobacter jejuni enteritis. He showed exaggerated tendon reflexes with abnormal reflex spread to other segments, and was initially diagnosed as having post-infectious myelitis. Nerve conduction studies showed motor axonal degeneration (the AMAN pattern), and increased soleus H-reflex amplitudes. His serum was positive for IgG antibodies to gangliosides GM1b and GalNAc-GD1a. He was treated with plasmapheresis, resulting in rapid recovery. Hyperreflexia was still present 12 months after onset when muscle strength was completely normal. This case provides further evidence that patients with AMAN can develop increased motor neuron excitability, and possible mechanisms for the hyperreflexia are discussed. PMID- 12084450 TI - Myoclonic involuntary movement associated with chronic manganese poisoning. AB - We report a 17-year-old man showing myoclonic involuntary movement (IVM) associated with chronic manganese (Mn) poisoning. The patient, a welder, showed myoclonic IVM mainly in the right upper and lower extremities, elevated levels of Mn in the blood and hair and high-intensity signals in the globus pallidus on T1 weighted MR images. Chelation therapy resulted in improvement of the myoclonic IVM and MRI abnormalities. This is the first report of Mn poisoning characterized by myoclonic IVM without parkinsonism. PMID- 12084452 TI - Development of transition state analogues of purine nucleoside phosphorylase as anti-T-cell agents. AB - Newborns with a genetic deficiency of purine nucleoside phosphorylase (PNP) are normal, but exhibit a specific T-cell immunodeficiency during the first years of development. All other cell and organ systems remain functional. The biological significance of human PNP is degradation of deoxyguanosine, and apoptosis of T cells occurs as a consequence of the accumulation of deoxyguanosine in the circulation, and dGTP in the cells. Control of T-cell proliferation is desirable in T-cell cancers, autoimmune diseases, and tissue transplant rejection. The search for powerful inhibitors of PNP as anti-T-cell agents has culminated in the immucillins. These inhibitors have been developed from knowledge of the transition state structure for the reactions catalyzed by PNP, and inhibit with picomolar dissociation constants. Immucillin-H (Imm-H) causes deoxyguanosine dependent apoptosis of rapidly dividing human T-cells, but not other cell types. Human T-cell leukemia cells, and stimulated normal T-cells are both highly sensitive to the combination of Imm-H to block PNP and deoxyguanosine. Deoxyguanosine is the cytotoxin, and Imm-H alone has low toxicity. Single doses of Imm-H to mice cause accumulation of deoxyguanosine in the blood, and its administration prolongs the life of immunodeficient mice in a human T-cell tissue xenograft model. Immucillins are capable of providing complete control of in vivo PNP levels and hold promise for treatment of proliferative T-cell disorders. PMID- 12084453 TI - A simple model to predict blood-brain barrier permeation from 3D molecular fields. AB - We report a four-component partial least squares discriminant analysis (PLS) model for the prediction of blood-brain barrier (BBB) permeation using descriptors derived from 3D molecular fields. The 3D fields were transformed by VolSurf into suitable 1D descriptors, which were correlated to the ratio of blood brain partitioning measured at steady state in rats (log C(brain)/C(blood)). The model so obtained sheds light on molecular properties influencing BBB permeation. It can also be used in the virtual screening of new chemicals. PMID- 12084454 TI - Control of alternative splicing by antisense oligonucleotides as a potential chemotherapy: effects on gene expression. AB - Expression of alternatively spliced mRNA variants at specific stages of development or in specific cells and tissues contributes to the functional diversity of the human genome. Aberrations in alternative splicing were found as a cause or a contributing factor to the development, progression, or maintenance of various diseases including cancer. The use of antisense oligonucleotides to modify aberrant expression patterns of alternatively spliced mRNAs is a novel means of potentially controlling such diseases. However, to utilize antisense oligonucleotides as molecular chemotherapeutic agents, the global effects of these molecules need to be examined. The advent of gene expression array technology has now made it possible to simultaneously examine changes that occur in the expression levels of several thousand genes in response to antisense treatment. This analysis should help in the development of more specific and efficacious antisense oligonucleotides as molecular therapeutics. PMID- 12084455 TI - Homeostatic control of uridine and the role of uridine phosphorylase: a biological and clinical update. AB - Uridine, a pyrimidine nucleoside essential for the synthesis of RNA and bio membranes, is a crucial element in the regulation of normal physiological processes as well as pathological states. The biological effects of uridine have been associated with the regulation of the cardio-circulatory system, at the reproduction level, with both peripheral and central nervous system modulation and with the functionality of the respiratory system. Furthermore, uridine plays a role at the clinical level in modulating the cytotoxic effects of fluoropyrimidines in both normal and neoplastic tissues. The concentration of uridine in plasma and tissues is tightly regulated by cellular transport mechanisms and by the activity of uridine phosphorylase (UPase), responsible for the reversible phosphorolysis of uridine to uracil. We have recently completed several studies designed to define the mechanisms regulating UPase expression and better characterize the multiple biological effects of uridine. Immunohistochemical analysis and co-purification studies have revealed the association of UPase with the cytoskeleton and the cellular membrane. The characterization of the promoter region of UPase has indicated a direct regulation of its expression by the tumor suppressor gene p53. The evaluation of human surgical specimens has shown elevated UPase activity in tumor tissue compared to paired normal tissue. PMID- 12084456 TI - Sequence-specific interactions of drugs interfering with the topoisomerase-DNA cleavage complex. AB - DNA-processing enzymes, such as the topoisomerases (tops), represent major targets for potent anticancer (and antibacterial) agents. The drugs kill cells by poisoning the enzymes' catalytic cycle. Understanding the molecular details of top poisoning is a fundamental requisite for the rational development of novel, more effective antineoplastic drugs. In this connection, sequence-specific recognition of the top-DNA complex is a key step to preferentially direct the action of the drugs onto selected genomic sequences. In fact, the (reversible) interference of drugs with the top-DNA complex exhibits well-defined preferences for DNA bases in the proximity of the cleavage site, each drug showing peculiarities connected to its structural features. A second level of selectivity can be observed when chemically reactive groups are present in the structure of the top-directed drug. In this case, the enzyme recognizes or generates a unique site for covalent drug-DNA binding. This will further subtly modulate the drug's efficiency in stimulating DNA damage at selected sites. Finally, drugs can discriminate not only among different types of tops, but also among different isoenzymes, providing an additional level of specific selection. Once the molecular basis for DNA sequence-dependent recognition has been established, the above-mentioned modes to generate selectivity in drug poisoning can be rationally exploited, alone or in combination, to develop tailor-made drugs targeted at defined loci in cancer cells. PMID- 12084457 TI - Self-association and unique DNA binding properties of the anti-cancer agent TAS 103, a dual inhibitor of topoisomerases I and II. AB - The objective of our study was to investigate the self-association and DNA binding properties of the DNA topoisomerases I (Topo I) and II (Topo II) dual inhibitor: 6-[[2-(dimethylamino)ethyl]amino]-3-hydroxy-7H-indeno[2,1-c]quinoline 7-one dihydrochloride (TAS-103), by means of 1H-NMR and 31P-NMR spectroscopy, structure computation techniques, thermal melting study, and UV-Visible spectroscopy. In aqueous solution, all chemical shifts of TAS-103 underwent upfield shifts depending with an increase in concentration. The two-dimensional (2D)-NMR spectra and structure computations indicated that TAS-103 self associated through pi-pi stacking and hydrophobic interactions of the aromatic chromophores. Thermal melting indicated that the binding of TAS-103 to DNA with a potency equal to that of ethidium bromide (EtBr). The UV-Visible spectra of TAS 103 titrated by several DNA exhibited hypochromic and hypsochromic effects. The 31P-NMR spectrum of the 6:1 TAS-103/d(CGCGAATTCGCG)(2) complex showed two broadening signals. 2D-NMR spectra of the 1:1 TAS-103/d(CGCGAATTCGCG)(2) complex indicated that the chemical shift differences of the DNA are very small. However, those of the terminal region are relatively large. The chemical shift differences of TAS-103 showed that the proton resonances except H2 underwent downfield shifts. From these observations, we conclude that TAS-103 binds to DNA by two modes. The major binding mode is on the surface (outside binding) and the minor binding mode by intercalation. PMID- 12084458 TI - Novel aspects of resistance to drugs targeted to dihydrofolate reductase and thymidylate synthase. AB - Drug resistance is often a limiting factor in successful chemotherapy. Our laboratory has been interested in studying mechanisms of resistance to drugs that are targeted to the thymidylate biosynthesis pathway especially those that target thymidylate synthase (TS) and dihydrofolate reductase (DHFR). We have used leukemia as a model system to study resistance to methotrexate (MTX) and colorectal cancer as the model system to study 5-fluorouracil (5-FU) resistance. In leukemias, we and others have shown that transport, efflux, polyglutamylation and hydrolase activities are major determinants of MTX resistance. We have further reported that some leukemic cells have an increase in DHFR gene copy number possibly contributing to the resistant phenotype. Recently, we have begun to study in detail the molecular mechanisms that govern translational regulation of DHFR in response to MTX as an additional resistance mechanism. Studies thus far involving colorectal tumors obtained from patients have focused predominantly on the predictive value of levels of TS expression and p53 mutations in determining response to 5-FU. Although the predictive value of these two measures appears to be significant, given the variety of resistance to 5-FU observed in cell lines, it is not likely that these are the only measures predictive of response or responsible for acquired resistance to this drug. The enzyme uridine cytidine monophosphate kinase (UMPK) is an essential and rate-limiting enzyme in 5-FU activation while dihydropyrimidine dehydrogenase (DPD) is a catabolic enzyme that inactivates 5-FU. Alterations in UMPK and DPD may therefore explain failure of 5-FU response in the absence of alterations in TS or p53. Transcription factors that regulate TS may also influence drug sensitivity. We have found that mRNA levels of the E2F family of transcription factors correlates with TS message levels and are higher in lung metastases than in liver metastases of colorectal cancers. Moreover, gene copy number of the E2F-1 gene appears to be increased in a significant number of samples obtained from metastases of colorectal cancer. We have also generated mutants of both DHFR and TS that confer resistance to MTX as well as 5-FU by random as well as site-directed mutagenesis. These mutants used alone or as fusion cDNAs of the mutants have proven to be useful in transplant studies where transfer of these mutant cDNAs to bone marrow cells have been shown to confer drug resistance to recipients. The fusion cDNAs of DHFR such as the DHFR-herpes simplex virus type 1 thymidine kinase (HSVTK) are also useful for regulation of gene expression in vivo using MTX as the small molecule regulator that can be monitored by positron emission tomography (PET) scanning or by optical imaging using a fusion construct such as DHFR-EGFP. PMID- 12084459 TI - Thymidylate synthase as a translational regulator of cellular gene expression. AB - Studies from our laboratory have shown that the folate-dependent enzyme, thymidylate synthase (TS), functions as an RNA binding protein. There is evidence that TS, in addition to interacting with its own TS mRNA, forms a ribonucleoprotein complex with a number of other cellular mRNAs, including those corresponding to the p53 tumor suppressor gene and the myc family of transcription factors. Using both in vitro and in vivo model systems, we have demonstrated that the functional consequence of binding of TS protein to its own cognate mRNA, as well as binding of TS to the p53 mRNA, is translational repression. Herein, we review current work on the translational autoregulatory control of TS expression and discuss the molecular elements that are required for the TS protein-TS mRNA interaction. TS may play a critical role in regulating the cell cycle and the process of apoptosis through its regulatory effects on expression of p53 and perhaps other cell cycle related proteins. Finally, the ability of TS to function as a translational regulator may have important consequences with regard to the development of cellular resistance to various anticancer drugs. PMID- 12084460 TI - Natural antisense (rTSalpha) RNA induces site-specific cleavage of thymidylate synthase mRNA. AB - The 3' untranslated region (UTR) of rTSalpha RNA is complementary (i.e., antisense) to human thymidylate synthase (TS) RNA. When HEp2 cells (human epidermoid carcinoma) progressed from late-log to plateau phase growth, ribonuclease protection assay (RPA) revealed an inverse correlation between the levels of rTSalpha RNA and TS mRNA, suggesting a possible effect of rTSalpha RNA on TS mRNA levels. HEp2 cells expressing a Tet-On transactivator were transiently co-transfected with pHook-1 and a construct containing rTSalpha (protein and antisense RNA), rTSalphaDelta3' (rTSalpha protein only), rTSalpha-3' (antisense RNA-luciferase) or luciferase. Transfected cells were selected and evaluated for the effects of induced transgene expression on TS mRNA. Induced expression of transfected rTSalpha or rTSalpha-3', but not rTSalphaDelta3' or luciferase, resulted in decreased TS mRNA levels as measured by RPA. These results demonstrated that the antisense region of rTSalpha RNA is necessary and sufficient for this down-regulation of TS mRNA. RPA for TS mRNA also showed the enhanced appearance of two partial-length protected fragments in rTSalpha or rTSalpha-3' transfected cells. RPA stringency evaluations and primer extension assays indicated that TS mRNA is cleaved in vivo in a site-specific manner. These data demonstrate that rTS gene expression likely plays a role in down-regulating TS through a natural RNA-based antisense mechanism. PMID- 12084461 TI - Induction of thymidylate synthase as a 5-fluorouracil resistance mechanism. AB - Thymidylate synthase (TS) is a key enzyme in the de novo synthesis of 2' deoxythymidine-5'-monophosphate (dTMP) from 2'-deoxyuridine-5'-monophosphate (dUMP), for which 5,10-methylene-tetrahydrofolate (CH(2)-THF) is the methyl donor. TS is an important target for chemotherapy; it is inhibited by folate and nucleotide analogs, such as by 5-fluoro-dUMP (FdUMP), the active metabolite of 5 fluorouracil (5FU). FdUMP forms a relatively stable ternary complex with TS and CH(2)THF, which is further stabilized by leucovorin (LV). 5FU treatment can induce TS expression, which might bypass dTMP depletion. An improved efficacy of 5FU might be achieved by increasing and prolonging TS inhibition, a prevention of dissociation of the ternary complex, and prevention of TS induction. In a panel of 17 colon cancer cells, including several variants with acquired resistance to 5FU, sensitivity was related to TS levels, but exclusion of the resistant variants abolished this relation. For antifolates, polyglutamylation was more important than the intrinsic TS level. Cells with low p53 levels were more sensitive to 5FU and the antifolate raltitrexed (RTX) than cells with high, mutated p53. Free TS protein down-regulates its own translation, but its transcription is regulated by E2F, a cell cycle checkpoint regulator. Together, this results in low TS levels in stationary phase cells. Although cells with a low TS might theoretically be more sensitive to 5FU, the low proliferation rate prevents induction of DNA damage and 5FU toxicity. TS levels were not related to polymorphisms of the TS promoter. Treatment with 5FU or RTX rapidly induced TS levels two- to five-fold. In animal models, 5FU treatment resulted in TS inhibition followed by a two- to three-fold TS induction. Both LV and a high dose of 5FU not only enhanced TS inhibition, but also prevented TS induction and increased the antitumor effect. In patients, TS levels as determined by enzyme activity assays, immunohistochemistry and mRNA expression, were related to a response to 5FU. 5FU treatment initially decreased TS levels, but this was followed by an induction, as seen with an increased ratio of TS protein over TS mRNA. The clear retrospective relation between TS levels and response now forms the basis for a prospective study, in which TS levels are measured before treatment in order to determine the treatment protocol. PMID- 12084462 TI - Structure-based studies on species-specific inhibition of thymidylate synthase. AB - Thymidylate synthase (TS) is a well-recognized target for anticancer chemotherapy. Due to its key role in the sole de novo pathway for thymidylate synthesis and, hence, DNA synthesis, it is an essential enzyme in all life forms. As such, it has been recently recognized as a valuable new target against infectious diseases. There is also a pressing need for new antimicrobial agents that are able to target strains that are drug resistant toward currently used drugs. In this context, species specificity is of crucial importance to distinguish between the invading microorganism and the human host, yet thymidylate synthase is among the most highly conserved enzymes. We combine structure-based drug design with rapid synthetic techniques and mutagenesis, in an iterative fashion, to develop novel antifolates that are not derived from the substrate and cofactor, and to understand the molecular basis for the observed species specificity. The role of structural and computational studies in the discovery of nonanalog antifolate inhibitors of bacterial TS, naphthalein and dansyl derivatives, and in the understanding of their biological activity profile, are discussed. PMID- 12084463 TI - A rationale for the clinical development of the thymidylate synthase inhibitor ZD9331 in ovarian and other solid tumours. AB - ZD9331 is an antifolate drug that potently and specifically inhibits thymidylate synthase (TS). In contrast with TS inhibitors such as raltitrexed, it cannot be polyglutamated, leading to antitumour activity independent of folylpolyglutamyl synthetase (FPGS) activity. The growth inhibition IC50 values for ZD9331 and raltitrexed were determined for a panel of 18 human tumour cell lines, that included six colon and six ovarian. The colon lines largely displayed overlapping sensitivities to both drugs with only one of the six lines being drug resistant. In contrast, the ovarian cell lines displayed non-overlapping sensitivities with four being highly resistant to raltitrexed and only one was cross-resistant to ZD9331. Studies were undertaken to explain these results. The colon and ovarian cell lines were characterised for TS activity, and TS and FPGS mRNA expression. TS activity correlated with sensitivity to ZD9331 (r=0.50; p=0.097) and raltitrexed (r=0.74; p=0.0063). Provided the data from the highly drug-resistant cell lines (BE and 41 M) were omitted, TS mRNA expression levels also correlated with ZD9331 (r=0.77; p=0.013) and raltitrexed IC50 (r=0.84; p=0.0031). FPGS mRNA expression correlated with higher sensitivity to raltitrexed relative to ZD9331 (higher ZD9331/raltitrexed IC50 ratios) (r=0.62; p=0.048). Similarly, cell lines with IC50 ratios>median expressed a 1.8-fold higher median level of FPGS mRNA (p=0.0087) compared with those with ratios or =30% of soluble intracellular proteins) through the construction of two-cistronic gene expression system. The solubilisation of recombinant growth hormones from inclusion bodies and subsequent oxidation to correctly folded monomeric form was also carried out. A combination of reverse phase HPLC and non-reducing SDS-PAGE was successfully applied to distinguish between reduced and oxidised forms of growth hormones. A moderate yield ( approximately 40% of starting material, with potential for upscaling), two-step purification process comprising of hydrophobic interaction and ion-exchange chromatographies was developed. The process eliminates the need for costly, laborious and time-consuming steps of ultrafiltration and dialysis, as reported earlier for the purification of many recombinant animal growth hormones. The biophysical, biochemical and functional analyses of purified refolded polypeptides showed that the hormones produced in this study were identical to natural pituitary bovine growth hormone. PMID- 12084479 TI - A new technique to prevent self-ligation of DNA. AB - The most widely used technique for preventing self-ligation (self-circularization and concatenation) of DNA is dephosphorylation of the 5'-end, which stops DNA ligase from catalyzing the formation of phosphodiester bonds between the 3' hydroxyl and 5'-phosphate residues at the DNA ends. The 5'-dephosphorylation technique cannot be applied to both DNA species to be ligated and thus, the untreated DNA species remains capable of self-ligation. To prevent this self ligation, we replaced the 2'-deoxyribose at the 3'-end of the untreated DNA species with a 2',3'-dideoxyribose. Self-ligation was prevented at the replaced 3'-end, while the 5'-phosphate remaining at the 5'-end permitted ligation with the 3'-hydroxyl end of the 5'-dephosphorylated DNA strand. We successfully applied this 3'-replacement technique to gene cloning, adapter-mediated polymerase chain reaction and messenger RNA fingerprinting. The 3'-replacement technique is simple and not restricted by sequence or conformation of the DNA termini and is thus applicable to a wide variety of methods involving ligation. PMID- 12084478 TI - Flax-retting by polygalacturonase-containing enzyme mixtures and effects on fiber properties. AB - Enzyme-retting of flax was accomplished via individual treatment with four polygalacturonase (PGase) containing solutions of various fungal sources and the resulting fibers were characterized. The retting solutions were equilibrated to contain 2.19 U of PGase activity as determined via a dinitrosalicylic acid (DNS) reducing sugar assay. As compared with the buffer control, treatment with the various enzyme solutions increased the yield of fine fibers. Treatment with Aspergillus niger PGase resulted in a 62% increase in fine fiber yield as compared with the buffer control and fiber strength did not statistically differ (P8h) followed by a slow decline; a characteristic of a non-Fickian diffusion process. Scanning electron microscopy studies suggested that the resultant release behavior was the outcome of the combined effects of the non-uniform distribution of the drug in the matrix and the apparent changes in the pores and surface characteristics of the microspheres. Comparison of release rate-time plots of dissolution data of marketed products with the newly developed dosage forms indicated the ability of the latter to sustain more zero order release. PMID- 12084507 TI - Reversible aggregation of lysozyme in a biodegradable amphiphilic multiblock copolymer. AB - Lysozyme-loaded poly(ethylene glycol terephthalate)-poly(butylene terephthalate) (PEGT/PBT) films were prepared using a water-in-oil emulsification solvent evaporation method. Infrared spectroscopic analysis of the dried films indicated the presence of non-covalent lysozyme aggregates in the polymer matrix. The use of methanol to enhance the drying rate of the films increased the relative amount of aggregates. Surprisingly, quantitative in-vitro release of fully active, non aggregated lysozyme was observed, indicating that lysozyme forms reversible aggregates during encapsulation in PEGT/PBT films. PMID- 12084508 TI - Pectinic acid, a novel excipient for production of pellets by extrusion/spheronisation: preliminary studies. AB - A very low soluble pectin-derivative (pectinic acid, degree of methoxylation 4%) was found to be well suited as an excipient for pelletisation by extrusion/spheronisation. Formulations containing pectinic acid and lactose in the following ratios were evaluated: 99/1, 80/20, 50/50 and 20/80. The capacity as an extrusion aid was found to be high; even formulations containing only 20% pectinic acid resulted in nearly spherical pellets. All pectinic acid pellets were mechanically stable, had an aspect ratio of approximately 1.15-1.20 and released 30-60% of a low solubility model drug within 15 min both in simulated gastric acid (0.1M HCl) and intestinal fluid (phosphate buffer pH 6.8). PMID- 12084511 TI - Cicerin and arietin, novel chickpea peptides with different antifungal potencies. AB - Two antifungal peptides with novel N-terminal sequences, designated cicerin and arietin were isolated from seeds of the chickpea (Cicer arietinum), respectively. Both peptides were adsorbed on Affi-gel blue gel and CM-Sepharose and exhibited a molecular weight of approximately 8.2 and 5.6 kDa, respectively. Arietin was more strongly adsorbed on CM-Sepharose than cicerin and manifested a higher translation-inhibiting activity in a rabbit reticulocyte lysate system and a higher antifungal potency toward Mycosphaerella arachidicola, Fusarium oxysporum and Botrytis cinerea. Both were devoid of mitogenic and anti-HIV-1 reverse transcriptase activities. PMID- 12084512 TI - The immunosuppressive activity of peptide fragments of vaccinia virus C10L protein and a hypothesis on the role of this protein in the viral invasion. AB - Our previous studies revealed that the 143-148 fragment of interleukin-1 receptor antagonist (IL-1 Ra) molecule with a Val-Thr-Lys-Phe-Tyr-Phe (VTKFYF) sequence inhibits the interleukin-1 (IL-1) interaction with its cellular receptor. The Val Thr-Arg-Phe-Tyr-Phe (VTRFYF) sequence of the 322-327 fragment of the C-terminal domain of vaccinia virus protein related to the C10L vaccinia gene shows a very high homology to the 143-148 IL-1 Ra fragment, suggesting a similar inhibitory activity. To test this suggestion, we investigated the inhibitory activity of a series of synthetic peptides derived from 316 to 327 fragment of C10L on the interaction of IL-1 with its receptor. We also tested the peptides for their influence on the humoral and cellular immune response. The results indicate that biological activities of the C10L fragments are similar to those obtained for respective fragments of IL-1 Ra. The C-terminal domain of C10L protein can be easily folded into spatial structure similar to the crystallographic one of IL-1 Ra. Based on the crystallographic structure of IL-1 Ra, we constructed a 3-D model of the C10L protein. According to the model, the Val(322)-Asn(328) sequence is localized on the surface of the molecule and, therefore, it may be involved in the interactions with receptors. Our results indicate that the C10L viral protein can play an important role in vaccinia virus evasion of the host immune system. It may consist in the blockade of IL-1 receptors by the C10L protein, a homologue of the IL-1 Ra. PMID- 12084513 TI - Effects of opiate ligands on intraspecific aggression in crickets. AB - In the cricket Gryllus bimaculatus, the opiate antagonist naloxone, 30 microg in 50 microl into hemolymph per animal, caused a release of intraspecific aggression in male-losers and in females. Naloxone had no significant effect on the aggression of winners and isolated males. The agonist of mu-opiate receptors DAGO, 45 microg, suppressed aggression in male winners and decreased the percentage and duration of contact fights between previously isolated males. The results suggest that, during social contacts, the activity of the endogenous opioid system may contribute to suppressing aggression in subordinate males, as well as in females. PMID- 12084515 TI - Antipeptide antibodies for detecting crab (Callinectes sapidus) molt-inhibiting hormone. AB - In crustaceans, the synthesis of ecdysteroid molting hormones is regulated by molt-inhibiting hormone (MIH), a neuropeptide produced by an eyestalk neuroendocrine system, the X-organ/sinus gland complex. Using sequence analysis software, two regions of the blue crab (Callinectes sapidus) MIH peptide were selected for antibody production. Two 14-mer peptides were commercially synthesized and used to generate polyclonal antisera. Western blot analysis revealed that each antiserum bound to proteins of the predicted size in extracts of C. sapidus sinus glands, and lysates of insect cells containing recombinant MIH. Thin section immunocytochemistry using either antiserum showed specific immunoreactivity in X-organ neurosecretory cell bodies, their associated axons and collaterals, and their axon terminals in the sinus gland. PMID- 12084514 TI - Potent antagonistic action of synthetic analogues of APGWGNamide, an antagonist of molluscan neuropeptide APGWamide. AB - Fifty-five kinds of analogues of APGWGNamide (Ala-Pro-Gly-Trp-Gly-Asn-NH2), which is an antagonist of molluscan neuropeptide APGWamide, were synthesized and their antagonistic activities were examined on two molluscan smooth muscles. Among all the analogues tested, on spontaneous contraction of the crop of the land snail, Euhadra congenita, APGWG(L-biphenylalanine, Bip)amide showed the most potent antagonistic activity and its potency was 50-100 times higher than that of APGWGNamide. Likewise, on phasic contraction of the anterior byssus retractor muscle (ABRM) of the sea mussel, Mytilus edulis, the effect of APGWG(D homophenylalanine, dHfe) was the most potent and showed 5-10 times stronger activity than that of APGWGNamide. In the tolerance test to known exo- and endopeptidases or the crop tissue homogenate, APGWGNamide was not only easily degraded by a proline-specific endopeptidase but also by the homogenate. Two kinds of potent antagonists were thus developed: APGWG(Bip)amide and APGWG(dHfe)amide, which will be useful tools for investigation of the function of APGWamide in the snail and the mussel, respectively. PMID- 12084517 TI - Effects of MCH/NEI in the striatum and interactions between them and with alpha MSH on IP3 levels. AB - In this study, we investigated: (a) the effect of melanocyte concentrating hormone (MCH) and neuropeptide glutamine (E)-isoleusine-(I) (NEI) on IP(3) production on an "in vitro" model using slices containing caudate putamen (CP) and accumbens nuclei (ACC); and (b) the interaction between these peptides and with alpha-melanocyte-stimulating-hormone (alpha-MSH) on the production of this second messenger. Only MCH at the highest dose studied (3.6 microM) increased the production of IP(3), whereas at the low concentration (0.6 microM) it did not affect IP(3) levels. NEI and alpha-MSH at both concentrations tested (0.6 and 3.6 microM), did not affect IP(3) production either. However, when NEI or alpha-MSH (at 3.6 microM) were added together with 3.6 microM MCH, the increase in the IP(3) content induced by this last peptide was blocked. PMID- 12084516 TI - Enzymatic activity of soluble and membrane tethered peptide pro-hormone convertase 1. AB - Pro-hormone convertases PC1 and PC2 perform endoproteolytic cleavages of precursors in peptide-containing secretory granules. PC1 and PC2 are soluble, secreted with bioactive peptides. Evolutionarily related PCs have membrane tethers, not secreted. We tethered PC1 to the transmembrane-cytoplasmic domains (CD) of a granule enzyme (peptidylglycine-alpha-amidating monooxygenase; PAM) and Golgi-localized PC8. The tethered PC1 is far more stable to elevated temperature and denaturants than soluble PC1, and more active. Both tethers allow PC1 to visit the cell surface transiently, cleaving soluble molecules outside the cell. Both membrane-bound PC1 chimeras cleave membrane PAM into soluble active fragments when PAM is expressed on adjacent cells. PMID- 12084518 TI - Differential responsiveness of CRF receptor subtypes to N-terminal truncation of peptidic ligands. AB - The role of the N-terminal domains of corticotropin-releasing factor (CRF) and CRF-like peptides in receptor subtype selectivity, ligand affinity and biological potency was investigated. Therefore, human CRF(12-41), human URP(12-38) and antisauvagine-30 (aSvg) were N-terminally prolonged by consecutive addition of one or two amino acids. The peptides obtained were tested for their binding affinities to rat CRF1 and murine CRF(2beta) receptor, and their capability to stimulate cAMP-release by HEK cells producing either receptor. It was observed that human CRF N-terminally truncated by eight residues was bound with high affinity to CRF2 receptor (Ki=5.4nM), whereas affinity for CRF1 receptor was decreased (Ki=250 nM). A similar shift of affinity was found with sauvagine (Svg) analogs. Truncation of human URP analogs did not affect their preference for CRF(2beta) receptor, but reduced their affinity. Changes in affinity were positively correlated with changes in potency. These results indicated that CRF1 receptor was more stringent in its structural requirements for ligands to exhibit high affinity binding than CRF(2beta) receptor. PMID- 12084519 TI - mu-Opioid receptor mRNA expression and immunohistochemical localization in the rat temporomandibular joint. AB - This study was undertaken to examine the presence and distribution of the mu opioid receptor (MOR) in the non-inflamed rat temporomandibular joint (TMJ) using non-radiographic in situ hybridization at the mRNA level and immunohistochemistry at the protein level. MOR mRNA and MOR-like immunoreactivity (MOR-LI) were found around the small blood vessels in the anterior part of the synovial membrane. The number of MOR mRNA signals in the anterior synovial membrane was significantly higher than that in the posterior part. Morphologically, MOR mRNA and MOR-LI were localized in amorphous materials considered to be nervous tissue, as well as some cell types considered to be macrophages, mast cells and endothelial cells. The present study showed the distribution of MOR in the rat TMJ synovial membrane and suggests that the opiate system plays an important role in endogenous analgesia in the TMJ. PMID- 12084520 TI - Differential antagonism of endomorphin-1 and endomorphin-2 supraspinal antinociception by naloxonazine and 3-methylnaltrexone. AB - To determine if different subtypes of mu-opioid receptors were involved in antinociception induced by endomorphin-1 and endomorphin-2, the effect of pretreatment with various mu-opioid receptor antagonists beta-funaltrexamine, naloxonazine and 3-methylnaltrexone on the inhibition of the paw-withdrawal induced by endomorphin-1 and endomorphin-2 given intracerebroventricularly (i.c.v.) were studied in ddY male mice. The inhibition of the paw-withdrawal induced by i.c.v. administration of endomorphin-1, endomorphin-2 or DAMGO was completely blocked by the pretreatment with a selective mu-opioid receptor antagonist beta-funaltrexamine (40 mg/kg), indicating that the antinociception induced by all these peptides are mediated by the stimulation of mu-opioid receptors. However, naloxonazine, a mu1-opioid receptor antagonist pretreated s.c. for 24h was more effective in blocking the antinociception induced by endomorphin-2, than by endomorphin-1 or DAMGO given i.c.v. Pretreatment with a selective morphine-6 beta-glucuronide blocker 3-methylnaltrexone 0.25mg/kg given s.c. for 25 min or co-administration of 3-methylnaltrexone 2.5 ng given i.c.v. effectively attenuated the antinociception induced by endomorphin-2 given i.c.v. and co-administration of 3-methylnaltrexone shifted the dose-response curves for endomorphin-2 induced antinociception to the right by 4-fold. The administration of 3-methylnaltrexone did not affect the antinociception induced by endomorphin-1 or DAMGO given i.c.v. Our results indicate that the antinociception induced by endomorphin-2 is mediated by the stimulation of subtypes of mu-opioid receptor, which is different from that of mu-opioid receptor subtype stimulation by endomorphin-1 and DAMGO. PMID- 12084521 TI - Family of hemorphins: co-relations between amino acid sequences and effects in cell cultures. AB - Hemorphins, i.e. endogenous fragments of beta-globin chain segment (32-41) LVVYPWTQRY(F) suppress the growth of transformed murine fibroblasts L929 cell culture, the effect is due to cytotoxicity and inhibition of cell proliferation. The contribution of cytotoxicity depends on the presence of Leu(32): VV hemorphins, except VV-hemorphin-4, exhibit cytotoxicity significantly higher than respective LVV-hemorphins. Decrease of cell number induced by hemorphins depend on the extent of N- and C-terminal degradation of hemorphins: VV-hemorphins in most cases are more active than LVV-, V-hemorphins, and hemorphins. In the group of VV-hemorphins the activity of VV-hemorphin-5 (valorphin) is significantly higher than of VV-hemorphin-7, VV-hemorphin-6, and VV-hemorphin-4, meaning that the presence of C-terminal Gln is important for suppressing of cell number. The amino acid sequence VVYPWTQ corresponding to valorphin was identified as important for manifestation of the both cytotoxic and antiproliferative effects. PMID- 12084522 TI - beta-Endorphin modulation of pressor response to hyperventilation in hypertensive patients. AB - After hyperventilation, systolic and diastolic blood pressure (BP) significantly decreased in 14 hypertensive patients (group 1), did not change in 9 (group 2) and increased in 8 (group 3). Basal BP, norepinephrine and dynorphin B levels were higher in group 1 than in groups 2 and 3. The decrease in BP after hyperventilation was associated with a decrease in plasma norepinephrine, Met enkephalin and dynorphin B and an increase in beta-endorphin. Naloxone abolished the hyperventilation-induced BP and norepinephrine decreases. Our findings indicate that hyperventilation may select hypertensive patients with different sympatho-adrenergic activity and that the increase in beta-endorphin reduces BP response to hyperventilation in patients with high sympatho-adrenergic tone. PMID- 12084523 TI - Tobacco smoke exposure and bombesin-like peptides in guinea pigs. AB - Bombesin-like peptides (BLPs) are associated with tobacco smoke (TS)-induced diseases. We sought to determine if acute TS exposure releases BLPs into the pulmonary circulation. Sensitized and non-sensitized guinea pigs were chronically exposed to TS or compressed air. Thereafter, the lungs were acutely challenged with TS while perfused. Perfusates were analyzed for BLPs. TS increased BLPs in non-sensitized guinea pigs. A separate study determined daily bombesin exposure increased lung cell counts but not airway hyperresponsivensess. TS exposure releases BLPs into the pulmonary circulation but can be modified by host factors and bombesin itself does not induce airway hyperresponsiveness. PMID- 12084524 TI - Neuropeptide Y Y1 and neuropeptide Y Y2 receptors in human cardiovascular tissues. AB - mRNA encoding the human NPY Y1 and NPY Y2 receptors were detected in cerebral, meningeal, and coronary arteries using reverse transcriptase-polymerase chain reaction (RT-PCR). In addition, the trigeminal and superior cervical ganglia were positive for both receptors. In some arteries and in SK-N-MC cells only mRNA encoding the NPY Y1 was detected. Besides the expected NPY Y1 PCR products, an additional 97 bp longer amplicon originating from an alternative splicing event was found in most tissues studied. Antibodies directed against the NPY Y1 receptor revealed immunostaining mainly in the smooth muscle layer of blood vessels whereas antibodies against the NPY Y2 receptor showed immunostaining in nerve cell bodies. PMID- 12084525 TI - Human bone marrow endothelial cells: a new identified source of B-type natriuretic peptide. AB - B-type natriuretic peptide (BNP) is a hormone mainly secreted by cardiac ventricle myocytes and which is increased in cardiac diseases. Moreover, BNP expression has been shown in various cell/tissue types. Six different human endothelial cell (EC) culture models arising from macro and microcirculation either primary cultures or cell lines were cultured and screened for BNP presence and secretion. All cell types expressed BNP mRNA while only the ECs arising from bone marrow stromal compartment secreted high amounts of BNP protein. This report is the first to identify ECs as a new source of BNP. However, BNP secretion is limited to a particular EC type. PMID- 12084526 TI - Age differences in neurokinin A and substance P from the hypothalamus, pituitary, pineal gland, and striatum of the rat. Effect of exogenous melatonin. AB - Previous data showed that aging of the central nervous system (CNS) is associated with widespread changes in tachykinin gene expression. However, there are no data about the possible role of exogenous melatonin in modulating the tachykinergic system during aging. The aim of this work was to analyze the age-dependent changes on neurokinin A (NKA) and substance P (SP) levels in hypothalamus, pituitary, pineal gland and striatum and the role of exogenous melatonin on these changes. We studied female rats at three different ages: 5-month-old (cyclic), 15 month-old (preacyclic) and 25-month-old (acyclic). Hypothalamic tachykinin levels increase when female rats reached acyclicity, this increase was blunted in acyclic-melatonin-treated rats. However, melatonin treatment in young cyclic rats resulted in significantly increased values as compared to controls. Pituitary NKA concentrations did no show age-dependent changes in control rats, however, in both, preacyclic and acyclic-melatonin-treated rats significantly increased values of pituitary NKA were found compared to controls. In the pineal gland, a marked decrease of NKA levels was observed in acyclic-control rats. Melatonin treatment did not alter this decrease. In the striatum, NKA and SP concentrations were significantly reduced in preacyclic- and acyclic-control rats compared to young cyclic rats, melatonin had no effect on striatal tachykinins. Our results indicate that melatonin may regulate tachykinin stores during aging mainly on structures of the neuroendocrine-reproductive axis. PMID- 12084527 TI - Selective tachykinin NK3-receptor agonists stimulate in vitro exocrine pancreatic secretion in the guinea pig. AB - The tachykinins, including substance P, neurokinin A and neurokinin B, are a mammalian peptide family that have documented motor, sensory and circulatory neurotransmitter functions in the gut. Little is known about their action on the exocrine pancreas. In this study we investigated the effects of PG-KII, a natural NK3-tachykinin receptor agonist, and senktide, a synthetic NK3-tachykinin receptor agonist, on amylase release from isolated pancreatic lobules of the guinea pig in comparison with the secretagogues carbachol, caerulein and substance P and the depolarizing agent KCl. When added to incubation flasks at various concentrations (from 10(-10) to 10(-6)M), PG-KII and senktide both caused a dose-dependent increase in amylase release from pancreatic lobules. PG-KII and senktide elicited a lower maximal response (7.5+/-0.8 and 8.1+/-0.6% of the total lobular amylase content) than carbachol (34.4+/-3.9%), caerulein (26.5+/-2.8%) and KCl (22.5+/-3.8%). Whereas atropine left PG-KII and senktide-stimulated secretion unaffected, the non peptide NK3 receptor antagonist SR 142801 significantly reduced the stimulant effect of PG-KII and senktide. PG-KII (10( 7)M) also slightly though significantly increased the response to lower concentrations of caerulein (10(-11) and 10(-10)M) and carbachol (10(-7) and 10( 6)M). These findings show that PG-KII and senktide are weak stimulants of exocrine pancreatic secretion that act directly on the acinar cells through NK3 receptors, without cholinergic involvement. We suggest also that the tachykininergic NK3 receptor system cooperates with the other known secretagogues in the control of pancreatic exocrine secretion. PMID- 12084529 TI - Anxiolytic-like effects of substance P fragment (SP(1-7)) in non-human primates (Callithrix penicillata). AB - The behavioral effects of the amino (N)-terminal fragment of substance P (SP(1 7)) on the marmoset (Callithrix penicillata) predator confrontation test of fear/anxiety were investigated. The test apparatus consisted of a figure-eight maze with three parallel arms interconnected at each extremity to a perpendicular arm. A taxidermized oncilla cat (Felis tigrina) was placed outside the maze facing one of its corners. Subjects were submitted to seven 30 min maze habituation trials (HTs), in the absence of the 'predator', and then to six 30 min treatment trials (TTs), in the presence of the 'predator', consisting of four doses of SP(1-7) (5, 50, 250 and 500 microg/kg; IP), saline and sham injection. SP(1-7) treatment reversed, in a dose-dependent way, the fear-induced avoidance behavior due to the predator's presence and increased the frequency of exploratory behaviors. Locomotor activity decreased during successive HTs, yet increased after all SP(1-7) treatments. These results indicate that systemic administration of SP(1-7) produces anxiolytic-like effects in marmosets tested in the predator confrontation model of fear/anxiety. PMID- 12084528 TI - Localization of calcitonin gene-related peptide receptors in rat gastric mucosa. AB - The location of calcitonin gene-related peptide (CGRP) receptors in the rat stomach has not been elucidated. It was recently reported that the CGRP receptor is formed when a calcitonin-receptor-like receptor (CRLR) and receptor activity modifying protein (RAMP) 1 are co-expressed on the cell membrane. The aim of this study was to determine the location and the role of CGRP receptors in the rat gastric mucosa. Gene expressions of CRLR and RAMP1 were investigated by Northern blot analysis, reverse transcription-polymerase chain reaction (RT-PCR), and in situ hybridization. Immunohistochemical stainings for CGRP, somatostatin, gastrin, and chromogranin A were performed. Gastric endocrine cells were collected by counterflow-elutriation and their responses to CGRP were studied. CRLR and RAMP1 mRNA was expressed mainly in small gastric epithelial cells in the pyloric glands. The mRNA expression had a similar distribution to that of D cells. In cultured gastric endocrine cells, CGRP enhanced somatostatin production, while it inhibited the secretion of histamine and gastrin. Our results suggest that CGRP receptors are expressed in D cells in the rat gastric mucosa and control production and secretion of somatostatin. PMID- 12084530 TI - Central LIF gene therapy suppresses food intake, body weight, serum leptin and insulin for extended periods. AB - Leukemia inhibitory factor (LIF) overexpression, induced by the intracerebroventricular (i.c.v.) injection of an recombinant adeno-associated viral vector encoding LIF (rAAV-LIF), resulted in a dose-dependent reduction in body weight (BW) gain, food intake (FI) and adiposity, evidenced by suppression of serum leptin and free fatty acids for an extended period in outbred adult female rats. A dose-dependent reduction in serum insulin levels and unchanged serum glucose, energy expenditure through thermogenesis as indicated by uncoupling protein-1 (UCP-1) mRNA expression in brown adipose tissue (BAT), and metabolism as indicated by serum T3 and T4, accompanied the blockade of weight gain. Thus, central rAAV-LIF therapy is a viable strategy to voluntarily reduce appetite and circumvent leptin resistance, a primary factor underlying age dependent weight gain and obesity in rodents and humans. PMID- 12084531 TI - Acute 3rd-ventricular amylin infusion potently reduces food intake but does not produce aversive consequences. AB - In this study, a conditioned taste aversion (CTA) paradigm was used to assess the possibility that 3rd-ventricular (i3vt) administration of the pancreatic hormone amylin produces aversive consequences that secondarily reduce food intake independently of the normal regulation of energy balance. After 1-h daily access to water for 7 days, rats were given 1-h access to a 0.15% saccharin solution, followed immediately by i3vt amylin (100 pmol) in one group (n=7) and i3vt CSF vehicle in another (n=7). As positive control for the formation of a CTA, a third group of seven rats received intraperitonial (i.p.) lithium chloride (LiCl). Saline was given i.p. to a fourth group (n=7) as control for i.p. LiCl. As expected, the LiCl rats exhibited a marked aversion to the saccharin in a subsequent two-bottle intake test. In contrast, although the 100 pmol i3vt amylin dose is substantially higher than that required to reduce food intake, no evidence of a CTA was observed in the rats that had received i3vt amylin. In summary, these data are consistent with the conclusion that acute i3vt amylin infusion does not reduce food intake by producing aversive consequences. PMID- 12084532 TI - Specificity and stability of a new PTH1 receptor antagonist, mouse TIP(7-39). AB - Parathyroid hormone 1 (PTH1) receptor antagonists might be of benefit in hypercalcemia of malignancy (HHM) and hyperparathyroidism. We previously identified bovine tuberoinfundibular peptide (7-39) (bTIP(7-39)) as a high affinity PTH1 receptor antagonist. Mouse TIP(7-39) is an antagonist (rPTH1 K(B)=44 nM, rPTH2=940 nM) that is more potent than other known PTH1 receptor antagonists: bTIP(7-39) (210 nM), PTH-related protein (PTHrP)(7-34) (640 nM), and bPTH(7-34) (>3000 nM). Plasma proteases slowly (t(1/2)=81 min) inactivated [125I] mTIP(7-39). Intravenous plasma [125I]mTIP(7-39) was bi-phasically cleared (radioactivity t(1/2)=2.9 min (70%) and 120 min (30%), binding activity t(1/2)=3.6 min (92%), and t(1/2)=21 min (8%)). Loss of unlabeled mTIP(7-39) (250 microg/kg i.v.) receptor binding was similar. mTIP(7-39)'s high-affinity should facilitate animal evaluation of effects of PTH1 receptor antagonism. PMID- 12084533 TI - Role of SA-Le(a) and E-selectin in metastasis assessed with peptide antagonist. AB - E-selectin ligand Sialyl-Lewis a (SA-Le(a)) carbohydrate is expressed on many carcinomas. Peptide mimicking SA-Le(a) (DLWDWVVGKPAG) was previously selected from a recombinant library by screening with monoclonal antibody (MAb) NS19-9. In this study, the residues critical for interaction with the NS19-9 were mapped using peptide array generated by substitution of various amino acid residues. The replacement of Trp 5 with Phe resulted in a change of peptide's secondary structure and increased binding with MAb and E-selectin, suggesting improved carbohydrate mimicry. Colonization of tumor cells expressing SA-Le(a) was blocked by the peptide and was completely abolished in E-selectin knock out mice. The data suggest the critical role of carbohydrate antigens and E-selectin in metastasis and that peptides mimicking carbohydrate antigens can function as antagonists of this process. PMID- 12084534 TI - Thymosin alpha1 inhibits mammary carcinogenesis in Fisher rats. AB - The effects of thymosin alpha1 (Talpha1) on mammary carcinogenesis was investigated in Fisher rats. Mammary carcinomas were observed 3 months after N nitrosomethylurea (NMU) injection (10mg, i.p.) into Fisher rats. Daily administration of Talpha1 (10 microg, s.c.) reduced mammary carcinoma incidence and prolonged survival time. Animals treated with exogenous Talpha1 had a significantly greater blood white cell density than control Fisher rats. These results suggest that Talpha1 prevents mammary carcinoma incidence as a result of stimulation of the immune system. PMID- 12084535 TI - In vitro effect of short-term exposure to two synthetic peptides, alone or in combination with clarithromycin or rifabutin, on Cryptosporidium parvum infectivity. AB - The viability of Cryptosporidium parvum after exposure to peptide antibiotics was studied by two different methods, a cell culture system and a double fluorogenic staining. The peptides KFFKFFKFF and IKFLKFLKFL exerted high cytotoxic effects on sporozoites, as demonstrated by cell cultures (complete inhibition after 60 min at 100 microg/ml) and flow cytometry (30% after 20 min at 100 microg/ml), but did not affect consistently the oocysts. Clarithromycin and rifabutin demonstrated less activity against sporozoites but higher activity against oocysts (30% after 180 min at 10 microg/ml). The combination between peptides and azithromycin or rifabutin exerted the highest activities. PMID- 12084537 TI - Energy storage and return prostheses: does patient perception correlate with biomechanical analysis? AB - The development and prescription of energy storage and return prosthetic feet in favor of conventional feet is largely based upon prosthetist and amputee experience. Regretfully, the comparative biomechanical analysis of energy storage and return and conventional prosthetic feet is rarely a motivation to either the technical development or clinical prescription of such devices. The development and prescription of prosthetic feet without supportive scientific evidence is likely due to the conflicting or non-significant results often presented in the scientific literature. Despite the sizeable history of comparative prosthetic literature and continued analysis of prosthetic components, the link between clinical experience and scientific evidence remains largely unexplored.A review of the comparative analysis literature evaluating energy storage and return and conventional prosthetic feet is presented to illustrate consistencies between the perceptive assessments and the objective biomechanical data. Results suggest that while experimental methodologies may limit the statistical significance of objective gait analysis results, consistent trends in temporal, kinetic, and kinematic parameters correlate well with perceptive impressions of these feet. These correlations provide insight to subtle changes in gait parameters that are deemed neither clinically nor statistically significant, yet are perceived by amputees to affect their preference for and performance of prosthetic feet during locomotion. Acknowledging and targeting areas of perceptive significance will help researchers develop more structured protocols for energy storage and return prosthesis evaluation as well as provide clinicians with information needed to enhance the appropriateness of their clinical recommendations. Expanding test environments to measure activities of perceived improvement such as high-velocity motions, stair ascent/descent, and uneven ground locomotion will provide a more appropriate assessment of the conditions for which energy storage and return prosthetic feet were designed. Concentrating research to specific test populations by age or amputation etiologies can overcome statistical limitations imposed by small study samples. Finally, directing research toward the areas of gait adaptation, heel performance, and the temporal release of energy in energy storage and return feet may reinforce the selection and utilization of advanced prosthetic components. These enhancements to current biomechanical analyses may serve to reduce the boundaries of perceptive significance and provide clinicians, designers, and researchers with the supportive data needed to prescribe, design, and evaluate energy storage and return prosthetic feet. PMID- 12084538 TI - Relationship between pain and vertebral motion in chronic low-back pain subjects. AB - OBJECTIVES: To investigate the relationship between intervertebral motion, intravertebral deformation and pain in chronic low-back pain patients. DESIGN: This study measured vertebral motion of the lumbar spine and associated pain in a select group of chronic low-back pain patients as they performed a standard battery of motions in all planes. BACKGROUND: Numerous studies have demonstrated that individuals with low-back pain have impaired spinal motion, yet few studies have examined the specific relationship between pain and motion parameters. Although it is accepted that the pain in mechanical low-back patients is due to specific spinal motions, no studies have related specific motions to pain measures. METHODS: Percutaneous intra-pedicle screws were placed into the right and left L4 (or L5) and S1 segments of nine chronic low-back pain patients. The external fixator frame was removed following the clinical external fixation test. The 3D locations of the pedicle screws and the level of pain were recorded as the subjects performed a battery of motions. The relationship between the pain and motion parameters was assessed using linear discriminant analysis and neural network models. RESULTS: The neural network model showed a strong relationship between observed and predicted pain (R(2)=0.997). The discriminant analysis showed a weak relationship (R(2)=0.5). CONCLUSIONS: Vertebral motion parameters are strongly predictive of pain in this select group of chronic low-back pain patients. The nature of the relationship is nonlinear and involves interactions; neural networks are able to effectively describe these relationships. RELEVANCE: Specific patterns of intervertebral motion and intravertebral deformation result in pain in chronic low-back pain patients. This substantiates the mechanical back pain aetiology. PMID- 12084539 TI - Examination of the flexion relaxation phenomenon in erector spinae muscles during short duration slumped sitting. AB - OBJECTIVE: The purpose of this study was to examine the myoelectric activity of the erector spinae muscles of the back in order to determine if the flexion relaxation phenomenon occurs in seated forward flexion or slumped postures. BACKGROUND: The flexion relaxation phenomenon during standing forward flexion is well documented. However, flexion relaxation in seated forward flexion has not been studied. It is possible that flexion relaxation could be linked with low back pain that some individuals experience during seated work. METHODS: Twenty two healthy subjects (11 males, 11 females) participated in the study. Surface electromyography was used to measure the level of muscle activity at the thoracic and lumbar levels of the erector spinae muscles. An electromagnetic tracking device measured the three-dimensional movement of the lumbar spine. Five trials each of standing and seated forward flexion were performed. RESULTS: A slumped sitting posture yielded flexion relaxation of the thoracic erector spinae muscles, whereas the lumbar erector spinae muscle group remained at relatively constant activation levels regardless of seated posture. Thoracic erector spinae silence occurred at a smaller angle of lumbar flexion during sitting than the flexion relaxation angle observed during standing flexion relaxation. CONCLUSIONS: Since the myoelectric activity of the lumbar erector spinae did not increase, it is likely that the passive tissues of the vertebral column were loaded to support the moment at L4/L5. Ligaments contain a large number of free nerve endings which act as pain receptors and therefore could be a potential source of low back pain during seated work. RELEVANCE: Examination of flexion relaxation during seated postures may provide insight into the association between low back pain and seated work. PMID- 12084540 TI - Influence of age on cervicothoracic spinal curvature: an ex vivo radiographic survey. AB - OBJECTIVE: To define the post-mortem cervicothoracic spinal curvature relative to age. DESIGN: Spinal curvature assessment of lateral cervicothoracic radiographs. BACKGROUND: A late consequence of age is the progressive accentuation of spinal curvatures, particularly the thoracic kyphosis. Little is known about the influence of the kyphosis on the alignment of the cervical spine. METHOD: One hundred and seventy two lateral spinal radiographs (113 males, 59 females) were analysed using two procedures: (1) sagittal curve deformation angles were derived, according to the method of Cobb, for thoracic (T1-T12), cervical (C2-C7) and cervicothoracic junctional regions (C6-T4); and (2) the cervicothoracic curvatures were digitised (C2-T12), to derive the apex of both curves and the inflexion point. RESULTS: A significantly increasing thoracic spinal curvature was determined for both genders, with the mean apex of the kyphosis close to T6. The cervical lordosis tended to flatten with increasing age, particularly in males, with the cervical apex location shifting cranially. This association was significant in older males and females. The mean location of the cervicothoracic curve inflexion point moved from T3 towards C7-T1 with increasing age. CONCLUSION: The cervicothoracic spinal curvature undergoes progressive change through the lifespan with a subsequent cranial migration of the inflexion point between the thoracic kyphosis and cervical lordosis, accompanied by a similar shift in the cervical apex. RELEVANCE: Sensitive measures of spinal curvature have utility in determining changes attributed to age, deformity or trauma on cervicothoracic spinal alignment. The value of assessing the location of curve inflexion lies in the ability to quantify changes in the relationship between different regions of the human spine without problems associated with identifying specific vertebral landmarks. PMID- 12084542 TI - Computer assisted screw insertion into real 3D rapid prototyping pelvis models. AB - OBJECTIVE: Show the use of computer navigation in exact screw positioning in the different pelvic bones. BACKGROUND: Computer assisted pedicle screw insertion in the spine is an established procedure. Screw fixation is also used in highly difficult pelvic and hip surgery (arthroplasty revision surgery and tumor surgery). DESIGN: Insert as long screws as possible with computer navigation into the different bones of the pelvis and compare these results with a non-computer controlled method. METHODS: The computer navigation was done with the system of Medivision (Oberdorf, Switzerland), the software was SurgiGATE 2.1. Optically controlled spine instruments and a special calibrated drill were used. The screw insertion with and without computer navigation took place in seven real rapid prototyping pelvis models matched by pairs. Three screws were inserted into the Os ileum, one into the Os pubis and one into the Os ischium. The length of the inserted screws were measured and compared using routine statistic methods. RESULTS: The mean screw length with vs. without computer navigation was 8.9 vs. 5.7 cm in the Os ileum (P=0.0001), 6.0 vs. 4.2 cm in the Os pubis (P=0.01) and 4.3 vs. 3.9 cm in the Os ischium (not significant). CONCLUSIONS: The use of computer navigation allows for the insertion of longer screws into the bones of the pelvis (more exact positioning), which requires a more precise original point of entry and direction of the drill (vector). RELEVANCE: The insertion of fixation screws in highly difficult pelvic and hip surgery (revision arthroplasty, tumor surgery) are another field for the use of computer navigation. PMID- 12084541 TI - Influence of laptop computer design and working position on physical exposure variables. AB - OBJECTIVE: To evaluate the impact of two laptop designs (with or without palm rest) and two work situations (on desk or lap) on neck and upper limb posture, muscle activity and productivity. DESIGN AND METHODS: Eight healthy subjects performed a standardized typing task of 15 min duration. During the last 5 min of each test, the neck, upper arm and trunk postures were captured by a three dimensional video system, wrist motion was measured by a biaxial electrogoniometer and muscle activity of four neck and upper limb muscles was recorded. RESULTS: Only minor differences in postures, wrist positions and productivity were observed when comparing the two laptop designs in the same situation. Larger differences were found when comparing the two situations (desk or lap). In the desk situation, the subjects bent their heads forward less, had less backward trunk inclination and wrist extension, but more elevation of the upper arm. Higher electromyographic (EMG) levels in the trapezius and deltoid muscles and lower EMG levels in the wrist extensors were also found in the desk situation. CONCLUSIONS: Our findings do not favor one particular laptop design because only small differences in physical exposure were found. However, the workstation set up influenced the physical exposure variables, and was pinpointed as the main determinant to be considered when doing laptop work even-though no ideal situation was found. Greater physical (muscular and articular) constraints seem to be imposed to the shoulder region in the desk situation whereas the head neck and wrist segments appear to be more stressed in the lap situation. RELEVANCE: Laptop computers are often used although the physical exposure in laptop work and the impact of different laptop designs have not been systematically assessed. A better understanding of these factors may help formulate some recommendations for laptop users. PMID- 12084543 TI - Unintentional forces developed during isometric test of the shoulder. AB - OBJECTIVE: To study unintentional, spontaneous forces developed during isometric shoulder strength testing. DESIGN: An experimental study. BACKGROUND: During torque measurements, subjects may apply forces in directions other than requested, thereby obscuring the interpretation of the results. METHODS: A shoulder strength test was performed with a strength test device permitting measurements of force in the requested direction as well as perpendicular to it. Nine fishermen and nine welders performed isometric abduction and elevation with the arm held in various angles. Moreover four rotational postures were investigated. RESULTS: Spontaneous force deviations from requested direction were found in almost all posture tested, with errors--defined as 100 times the ratio of the magnitude of intentional torque to the magnitude of unintentional (erroneous) torque generated perpendicular to it--of more than 30% found for some postures. Abduction in 45 degrees horizontal flexion was better controlled than abduction in 90 degrees horizontal flexion. Detailed analysis indicated some groupwise differences between fishermen and welders. Also pairwise comparison of handiness indicated some differences. CONCLUSIONS: Considerable deviations from requested direction of action can arise during strength tests. Thus, one must either monitor these deviations or minimize them by a proper design of the examination to get relevant data. As the error can differ between certain subject categories depending on posture this indicate a new tool for discriminating between musculo-skeletal problems. RELEVANCE: Muscle strength devices must be used with care as their measurement setups may mask the truce muscle action of the patients. PMID- 12084544 TI - In vivo determination of subject-specific musculotendon parameters: applications to the prime elbow flexors in normal and hemiparetic subjects. AB - OBJECTIVE: This study aimed at estimating the musculotendon parameters of the prime elbow flexors in vivo for both normal and hemiparetic subjects. DESIGN: A neuromusculoskeletal model of the elbow joint was developed incorporating detailed musculotendon modeling and geometrical modeling. BACKGROUND: Neuromusculoskeletal modeling is a valuable tool in orthopedic biomechanics and motor control research. However, its reliability depends on reasonable estimation of the musculotendon parameters. Parameter estimation is one of the most challenging aspects of neuromusculoskeletal modeling. METHODS: Five normal and five hemiparetic subjects performed maximum isometric voluntary flexion at nine elbow positions (0 degrees -120 degrees of flexion with an increment of 15 degrees ). Maximum flexion torques were measured at each position. Computational optimization was used to search for the musculotendon parameters of four prime elbow flexors by minimizing the root mean square difference between the predicted and the experimentally measured torque-angle curves. RESULTS: The normal group seemed to have larger maximum muscle stress values as compared to the hemiparetic group. Although the functional ranges of each selected muscle were different, they were all located at the ascending limb of the force-length relationship. The muscle optimal lengths and tendon slack lengths found in this study were comparable to other cadaver studies reported in the literature. CONCLUSION: Subject-specific musculotendon parameters could be properly estimated in vivo. RELEVANCE: Estimation of subject-specific musculotendon parameters for both normal and hemiparetic subjects would help clinicians better understand some of the effects of this pathological condition on the musculoskeletal system. PMID- 12084545 TI - A biomechanical analysis of the rheumatoid index finger after joint arthroplasty. AB - OBJECTIVE: To determine the mechanisms responsible for the recurrence of ulnar drift after metacarpophalangeal joint arthroplasty in the rheumatoid hand. DESIGN: A three-dimensional biomechanical model of the index finger joints was used to predict the implant loads during several activities of daily living. BACKGROUND: Post-operative clinical evaluation of Sutter metacarpophalangeal prostheses shows a high incidence of fracture and recurrent deformity. METHODS: A six-component force transducer in conjunction with a six-camera motion analysis system were used to obtain kinematic and external loading data from eight patients with rheumatoid arthritis during several simulated activities. These data were used as input into a three-dimensional biomechanical model of the implant and interphalangeal joints of the index finger. Tendon lines of action and moment arms were obtained using a series of MRI scans and CAD modelling techniques. RESULTS: Implant forces were oriented in a radial and dorsal direction to resist the ulnarpalmarly pull of tendons associated with the metacarpophalangeal joint. CONCLUSIONS: The recurrence of ulnar drift is attributable to fatigue failure of the prostheses. After fracture the implant is unable to support the repetitive loading patterns experienced during activities of daily living. RELEVANCE: Understanding the mechanisms responsible for the recurrence of ulnar drift and implant failure is a step towards improving the prosthesis design, surgical procedures and ultimately the patient's prognosis. PMID- 12084546 TI - Measurement of plantar pressure distribution in hemiplegic children: changes to adaptative gait patterns in accordance with deficiency. AB - OBJECTIVE: A comparison of plantar pressure distribution in hemiplegic children with a healthy control group was performed to illustrate the link between the changing dynamics during the stance phase and the degree of deficiency. DESIGN: Twenty three healthy control subjects and two groups of six hemiplegic children with minor and major spasticity were tested. BACKGROUND: In previous studies, it was shown that the musculoskeletal dysfunction due to spasticity disturbed the hemiplegic gait pattern, for example reduced walking speed and step length. However, plantar pressure measurements which would help to understand the pathological gait mechanisms have not been studied. METHODS: The stance phase parameters measured included spatio-temporal data and the relative impulse measurements during consecutive gait cycles. The relative impulses under eight plantar areas were determined with an in-shoe plantar pressure measurement device. RESULTS: Each group had a specific plantar pressure distribution profile that varied with the degree of deficiency. The most significant differences were found beneath the midfoot, the first metatarsal head and the hallux. CONCLUSION: The neuromuscular disorders and foot deformities due to the cerebral lesion modified the contact of the foot with the ground, and led to specific plantar pressure distribution profiles. The equinovarus, with clawed-toe deformity due to greater spasticity, seemed to be an important factor in disturbances of the terminal stance phase. However, the spastic hemiplegic subjects seemed to adopt a gait pattern required for optimal stability. RELEVANCE: From previous work, comparable data were available only for the hemiparetic adult patients. In-shoe pressure data from spastic hemiplegic children can provide detailed information specific to each region of contact for the support of diagnosis, a clinical decision or the prescription of appropriate footwear, braces or othoses. Both peak pressure and local impulse have proven to be valuable for the understanding of foot function, lower extremity dysfunctions and walking strategies. PMID- 12084547 TI - The influence of fatigue on trunk muscle responses to sudden arm movements, a pilot study. AB - OBJECTIVE: To examine fatigue induced changes in trunk muscle latencies following trunk muscle fatigue. DESIGN: A repeated measures within subject design.Background. Trunk muscle responses to sudden movements is of interest in clinical biomechanics and motor control. METHODS: Electromyographic profiles were recorded from transversus abdominis (finewire), internal oblique, rectus abdominis and external oblique and longissimus at the level of the 3rd lumbar vertebrae bilaterally. Four asymptomatic subjects performed arm-raising task using a visual cue before and after an isometric fatiguing trunk extension task. RESULTS: Feed-forward responses were not detected in all muscles for every trial. In general, following fatigue trunk muscle onset latencies occur earlier (left, P=0.0016; right, P=0.0475). CONCLUSIONS: Trunk muscle fatigue alters anticipatory postural adjustments in normal subjects. It remains unclear if there is a pattern for specific muscles changes between individuals and if these are reflected in individuals with low back pain. RELEVANCE: Trunk muscle fatigue and altered trunk muscles latencies to movement perturbations have been associated with low back pain. These findings suggest that there may be a link between centrally mediated response to isometric muscle fatigue and anticipatory motor control strategies. PMID- 12084549 TI - Prolyl 4-hydroxylase inhibitor is more effective for the inhibition of proliferation than for inhibition of collagen synthesis of rat hepatic stellate cells. AB - The aim of this study was to investigate whether a prolyl 4-hydroxylase inhibitor (HOE 077) prevents the proliferation and collagen synthesis of rat hepatic stellate cells (HSCs). Rat HSCs were isolated and cultured with 100, 500, 1000 or 2000 &mgr;g/ml of HOE 077 with or without hepatocytes. After 4 day culture, the cell cycle of HSCs was examined by flow cytometry along with messenger RNA expression of procollagen type I. After 4 days of culture, HSCs had DNA synthesis and a high concentration (2000 &mgr;g/ml) of HOE 077 significantly reduced this DNA synthesis. However, HOE 077 incubated with hepatocytes could significantly reduce mitosis at a lower concentration of HOE077 (500 &mgr;g/ml) without significant reduction of alpha(2)(I) procollagen mRNA expression. These results indicate that the prolyl 4-hydroxylase inhibitor is more effective for the inhibition of proliferation than for inhibition of collagen synthesis of HSCs. PMID- 12084548 TI - The effect of shoulder movements on thoracic spine 3D motion. AB - OBJECTIVE: To investigate whether there is any repeatable coupling pattern of upper thoracic movements, during right arm elevation in sagittal and scapular plane. METHODS: Upper thoracic movement patterns (T2-T7) were examined during right arm elevation, in the sagittal and scapular plane using an electromagnetic device. The participants were 25 asymptomatic woman aged between 45 and 64 years. RESULTS: The majority of the participants performed an ipsilateral coupling pattern between lateral flexion and rotation which was repeatable and comparable for both arm elevation planes. Ipsilateral coupling of right lateral flexion and rotation in association with extension occurred in 23 subjects during arm elevation in sagittal plane, and in 19 subjects during arm elevation in scapular plane. CONCLUSIONS: Analysis revealed that the participants demonstrated a repeatable upper thoracic movement pattern within and between subjects, during arm elevation in sagittal and scapular planes. RELEVANCE: The biomechanical relationship between the arm and the thoracic spine is important to clinical practice because of the contribution of spinal to shoulder movements and vice versa, in the recognition, prevention and restoration of musculoskeletal function. PMID- 12084550 TI - Novel arterial infusion chemotherapy using cisplatin, 5-fluorouracil, and leucovorin for patients with advanced hepatocellular carcinoma. AB - Advanced hepatocelluar carcinoma (HCC) has a poor prognosis. In this study, the authors evaluated the efficacy of chemotherapy using cisplatin (CDDP), 5 fluorouracil (5-FU), and leucovorin (LV), comparing our regimen with chemotherapy using CDDP and 5-FU. Nineteen patients with advanced HCC were treated by arterial infusion of a chemotherapeutic agent via a subcutaneously implanted injection port. In Group A (n=9), one course of chemotherapy consisted of the daily administration of CDDP (10 mg/1 h, on 5 days) and LV (12 mg/10 min, on 5 days) followed by 5-FU (250 mg/5 h, on 5 days). In Group B (n=10), except for the administration of LV, the same regimen was employed. This course was repeated each week for 4 weeks. In Group A, two patients showed a complete response (CR), and the other three showed a partial response (PR). In Group B, two patients showed PR. The response rate (CR+PR/all cases) in Group A was significantly higher than that in Group B (56 vs. 20%; P=0.022). The 1- and 2-year survival rates of Group A (66.7, 44.4%) were significantly higher than those of Group B (10, 0%) (P=0.033). These results suggest that our regimen may be useful in treating patients with advanced HCC. PMID- 12084551 TI - Population-based study of hepatitis C virus infection and hepatocellular carcinoma in western Japan. AB - Hepatitis C virus (HCV) infection is a major health problem in Japan. This infection is highly prevalent in subjects with chronic liver disease and is strongly associated with hepatocellular carcinoma. Epidemiological studies conducted by the Japanese Ministry of Health and Welfare showed that the mortality rate associated with hepatocellular carcinoma (HCC) was high in several prefectures in western Japan. In 1990, Saga Prefecture on the island of Kyushu in western Japan reported the highest HCC related mortality rate. Here, we report the results of a population-based survey in Saga Prefecture, where prevalence of HCV infection and mortality rate of HCC was assessed in the general population. From 1992 to 1997, the prevalence of HCV infection was assessed in the general population of 47 districts in Saga Prefecture. Among the 161307 subjects (52590 men, 108357 women, older than 30 years) examined, 13129 (8.1%) reacted positively to HCV antibody (anti-HCV). The prevalence of HCV-Ab reactivity was highest in subjects over 60 years of age. Reactivity increased from 3.6% in subjects 30-49 years old to 11% in those >50 years old. Highly significant differences were observed among the 45 districts in seropositive rates for anti-HCV, with the range being from 0.8 to 20.0%. We evaluated the association between the prevalence of anti-HCV reactivity and age-adjusted death rate from HCC in the general population of these districts, and detected a significant association (Pearson' s correlation coefficient=0.721, P<0.0001, Y=1.86X+16.1). In conclusion, these observations indicated that the outbreak of HCV in this area was a major cause of HCC in the population. PMID- 12084552 TI - An experimental study on Nd-YAG laser induced thermotherapy. Its possible application of the laser irradiation for therapy of hepatocellular carcinoma. AB - It has been reported that the local thermotherapy is useful in treatment of hepatocelluar carcinoma complicated with severe liver cirrhosis (LC). We studied the effect of laser beam generated by a newly developed Nd-YAG laser system on liver tissue of piglet. The system was composed of their quartz bare fiber with a cone-shaped tip. After liver was exposed by incision of abdomen under general anesthesia, the fiber was inserted into liver tissue and then the laser beam was generated. The power, duration and frequency of irradiation were varied. The coagulated area was inspected measured and examined histologically. The system produced an oval-shaped coagulated area in front of the needle tip. Furthermore, the volume of tissue coagulated was correlated with the total output of laser, enabling us to estimate the coagulated volume based on the laser output applied. Our findings indicated that Nd-YAG laser-induced thermotherapy would be successfully applied for the clinical treatment of HCC. PMID- 12084553 TI - Analysis of natural killer cells and interleukin-15 in patients with acute and fulminant hepatitis. AB - To clarify the role of natural killer (NK) cells in acute hepatitis (AH) and fulminant hepatitis (FH), we measured the prevalence of CD16+ and CD57+ NK cells and the serum levels of IL-15, an inducer of NK cell activity. We analyzed 14 healthy subjects, 30 AH patients, and 13 FH patients for the percentages of CD16+ and CD57+ cells in peripheral blood lymphocytes and the serum IL-15 levels. In the AH patients with HAV, the percentage of CD16+ cells was significantly increased compared to healthy subjects, but that of CD57+ cells did not differ between the groups. The mean values of IL-15 levels of AH with HAV and unknown etiology, as well as of FH were higher than those of healthy subjects. Four subacute type FH (S-FH) patients had an increase in serum IL-15 levels, as well as the percentage of CD16+ cells throughout the course of their illness. However, in four acute type FH patients, this phenomenon was not observed. Our data suggest that NK cell immunity appears to be important in patients with AH secondary to HAV infection. In addition, the number of IL-15 and NK cells increased in the later stages of the illness, which may have contributed to the pathogenesis of S-FH. PMID- 12084554 TI - Activation and maturation of antigen-presenting dendritic cells during vaccine therapy in patients with chronic hepatitis due to hepatitis B virus. AB - BACKGROUND/AIMS: The function and phenotype of antigen-presenting dendritic cells (DC) have been checked in patients with chronic hepatitis (CH) undergoing vaccine therapy to have insight about the mechanism of this immune therapy. SUBJECTS AND METHODS: Eleven patients with CH due to hepatitis B virus (HBV) were injected with vaccine containing 20 &mgr;g of hepatitis B surface antigen (HBsAg) once at every 2 weeks for 12 consecutive times. The serum interluekin-12 levels were estimated and the function and phenotype of peripheral blood DC were checked before, during and after the vaccine therapy. RESULTS: Sustained normalization of DNA-polymerase activity and alanine aminotransferase were seen in six patients (complete responders [CR]) due to vaccine therapy. The levels of serum interleukin-12 (P<0.01), the stimulatory capacity of peripheral blood DC (P<0.05) and the numbers of CD83-positive mature DC and CD86-positive activated DC (P<0.05) were significantly increased due to vaccination in CH-B patients, especially in younger patients. CONCLUSIONS: A role of DC during vaccine therapy is shown. This study also indicates that more effective vaccine therapy can be developed by upregulating the function of DC in situ in younger patients. PMID- 12084555 TI - Quantitative analysis of wild-type and precore mutant hepatitis B virus in carriers. AB - In the present study, we have analyzed the amount of precore wild-type hepatitis B virus (HBV) (wild-type) and precore mutant HBV (nt 1896: G-->A) (precore mutant) of HBV carriers; 31 asymptomatic healthy carriers (ASC) and 28 patients with chronic hepatitis (CH). Wild-type and precore mutant were quantified using sensitive and specific quantification methods: competitive wild-type-sequence specific assay and competitive mutation-site-specific assay with different sets of specific primers and internal controls. Median serum levels of wild-type and precore mutant were 9.60 and 8.60 log copies/ml (median percentages of precore mutant in total HBV-DNA: 11.7%) in HBeAg(+) ASC, 8.48 and 8.00 (33.3%) in HBeAg(+) CH, and 6.30 and 6.85 (84.7%) in anti-HBe(+) CH, respectively, showing higher levels of the relative amount of precore mutant to wild-type along with HBeAg/anti-HBe status. Only precore mutant, but not wild-type was detected in anti-HBe(+) ASC. Although median percentages of precore mutant at the anti-HBe(+) ASC and CH stages were much higher than those at the HBeAg(+) ASC and CH stages, a substantial amount of precore mutant was found even at the HBeAg(+) stages. Existence of a substantial amount of precore mutant even in HBeAg(+) ASC suggests that the occurrence of precore mutant is not always closely associated with seroconversion from HBeAg to anti-HBe. PMID- 12084556 TI - Clinical spectrum of acute sporadic hepatitis E and possible benefit of glycyrrhizin therapy. AB - The aim of the present study was to record the spectrum of sporadic hepatitis due to hepatitis E virus infection with special reference to moderate and severe liver disease, described as sub-acute hepatitis. Further, efficacy of glycyrrhizin therapy was studied as an open trial. Sixty-two consecutive patients were registered for the study. The clinical and laboratory profile of the patients was recorded on a preplanned proforma. Moderate and severe hepatitis was arbitrarily defined on the basis of clinical symptoms and serum bilirubin (total) of 10-15 mg% and 16 mg% or higher, respectively, at the time of presentation. It was noted that 22 (36.1%) of acute sporadic hepatitis E patients had moderate or severe liver disease. Glycyrrhizin was administered to these 22 patients by intravenous (IV) route in the dose of 60 ml daily. Therapy was tapered and stopped once significant clinical and biochemical improvement was noted. All patients showed clinical improvement by the seventh day of therapy. Total bilirubin was reduced by 68.9% by the end of 2 weeks of treatment and at this time, reduction in AST and ALT levels was to the tune of 94 and 97%, respectively. Normalization of AST and ALT levels was recorded in 19 patients (86.4%) and total bilirubin in 13 (59.1%) patients within 30 days of commencement of therapy. There were no side effects of IV glycyrrhizin therapy. It is concluded from the results of the present study that over one-third patients with acute sporadic hepatitis E in India have either moderate or severe liver injury. IV glycerrhizin therapy in this group of patients is well tolerated and effective. PMID- 12084557 TI - Transient biochemical response in interferon therapy decreases the development of hepatocellular carcinoma for five years and improves the long-term survival of chronic hepatitis C patients. AB - The development of hepatocellular carcinoma (HCC) was significantly reduced in both sustained responders (SR) and transient biochemical responders (TR) in chronic hepatitis C (CH-C) patients who received interferon (IFN) therapy. However, the long-term clinical outcome of TR remains unclear. One thousand three hundred and seventy CH-C Japanese patients who received IFN therapy and 54 control cirrhotic patients were enrolled. TR were defined as those patients who showed a normal serum alanine aminotransferase level (<==30 IU/l) at the end of therapy and then relapsed. Mean follow-up period was 5.6 years (6.1 years in 48 cirrhotic patients) in the IFN group and 8.3 years in the 54 control cirrhotic patients. HCC was detected in 114 patients in the IFN group among whom 4 were in the 425 SR, 21 were in the 359 TR and 89 were in the 586 non-responders (NR). The cumulative incidence of HCC was significantly (P=0.0001) inhibited in both SR and TR compared with NR. Its inhibitory effect in TR was within 5 years. Platelet count did not significantly decrease for 2-4 years after IFN therapy in TR, but it significantly decreased in NR 2 years after IFN therapy. The cumulative survival in both SR and TR was significantly higher than NR (SR vs NR; P=0.0001, TR vs NR; P=0.0305). These results indicate that IFN therapy lowers the rate of the progression of HCC and improves the long-term survival even in CH-C patients who transiently respond to IFN therapy. PMID- 12084558 TI - Expression of the hepatocyte canalicular multidrug resistance protein (MRP2) in primary biliary cirrhosis. AB - The canalicular multidrug resistance protein 2 (MRP2; gene symbol: ABCC2) mediates ATP-dependent biliary excretion of organic anions such as bilirubin diglucuronide, glutathione conjugates and sulfated and glucuronidated bile salts. In chronic cholestatic liver diseases, the biliary excretion of cholephilic organic anions is impaired. While the underlying transport defects have been studied in rat models of cholestasis, little is known about the molecular basis of impaired organic anion excretion in human cholestatic liver disease. Our aim, therefore, was to analyze expression of MRP2 in patients with primary biliary cirrhosis (PBC), a chronic cholestatic liver disease characterized by progressive destruction of small intrahepatic bile ducts. Four patients with PBC stages III (n=1) and IV (n=3) were compared with three non-cholestatic patients with alcoholic liver disease, idiopathic liver cirrhosis and cirrhosis from chronic hepatitis C. Immunohistochemistry was performed on paraffin-embedded tissue slides using a monoclonal antibody to MRP2. MRP2 was detected at the canalicular hepatocyte membrane of all patients. In two PBC patients (stages III and IV, respectively), the degree of immunostaining was comparable with controls, whereas in two other PBC patients with stage IV disease immunostaining was decreased. We conclude that MRP2 expression decreases with progressive cholestasis in PBC. PMID- 12084559 TI - A method for spike sorting and detection based on wavelet packets and Shannon's mutual information. AB - Studying the dynamics of neural activity via electrical recording, relies on the ability to detect and sort neural spikes recorded from a number of neurons by the same electrode. We suggest the wavelet packets decomposition (WPD) as a tool to analyze neural spikes and extract their main features. The unique quality of the wavelet packets-adaptive coverage of both time and frequency domains using a set of localized packets, facilitate the task. The best basis algorithm utilizing the Shannon's information cost function and local discriminant basis (LDB) using mutual information are employed to select a few packets that are sufficient for both detection and sorting of spikes. The efficiency of the method is demonstrated on data recorded from in vitro 2D neural networks, placed on electrodes that read data from as many as five neurons. Comparison between our method and the widely used principal components method and a sorting technique based on the ordinary wavelet transform (WT) shows that our method is more efficient both in separating spikes from noise and in resolving overlapping spikes. PMID- 12084560 TI - Extracellular potentials in low-density dissociated neuronal cultures. AB - The detection of extracellular potentials by means of multi-electrode arrays (MEA) is a useful technique for multi-site long-term monitoring of cultured neuronal activity with single-cell resolution. To optimize the geometry of the MEA it is advantageous to localize the cellular compartments that constitute the generators of these signals. For this purpose, an in vitro technique for the detection of extracellular signals with subcellular resolution has been developed. It makes use of easy-to-manufacture large-tip pipettes, monitoring of electrode-cell gap resistance for precise electrode positioning and low-density (100 cells/mm(2)) dissociated hippocampal cultures. Negative monophasic extracellular spikes, typically 60 microV, were measured over putative axonal processes and monophasic, biphasic and triphasic signals were recorded over the soma. A compartmental simulation suggests that different somatic conductance densities of Na(+) (1-10 mS/cm(2)) and K(+) (5-10 mS/cm(2)) channels can produce characteristic somatic extracellular potentials, with a variety of shapes similar to those observed experimentally. PMID- 12084562 TI - MEA-Tools: an open source toolbox for the analysis of multi-electrode data with MATLAB. AB - Recent advances in electrophysiological techniques have created new tools for the acquisition and storage of neuronal activity recorded simultaneously with numerous electrodes. These techniques support the analysis of the function as well as the structure of individual electrogenic cells in the context of surrounding neuronal or cardiac network. Commercially available tools for the analysis of such data, however, cannot be easily adapted to newly emerging requirements for data analysis and visualization, and cross compatibility between them is limited. In this report we introduce a free open source toolbox called microelectrode array tools (MEA-Tools) for the analysis of multi-electrode data based on the common data analysis environment MATLAB (version 5.3-6.1, The Mathworks, Natick, MA). The toolbox itself is platform independent. The file interface currently supports files recorded with MCRack (Multi Channel Systems, Reutlingen, Germany) under Microsoft Windows 95, 98, NT, and 2000, but can be adapted to other data acquisition systems. Functions are controlled via command line input and graphical user interfaces, and support common requirements for the analysis of local field potentials, extracellular spike activity, and continuous recordings, in addition to supplementary data acquired by additional instruments, e.g. intracellular amplifiers. Data may be processed as continuous recordings or time windows triggered to some event. PMID- 12084561 TI - Evaluation of brain toxicity following near infrared light exposure after indocyanine green dye injection. AB - Indocyanine green (ICG) has excellent safety records and is widely used in medical diagnosis. Recently, a new method has been developed to estimate cerebral blood flow (CBF) using ICG in combination with near-infrared spectroscopy (NIRS). The new technique may be of wide clinical interest, as it is noninvasive and easy to perform at the bedside in stroke patients. Additionally, ICG with the use of specific wavelength lasers is documented to be effective in photodynamic therapy (PDT). Under normal conditions ICG does not cross the intact blood brain barrier (BBB). However, in patients with brain injuries where the BBB may be disturbed, ICG could accumulate in brain parenchyma and in combination with NIR-light exposure, phototoxicity could occur. The aim of the present study was to examine the possible toxicity of ICG in combination with NIRS in a specific setting for CBF measurements. In five rats with mannitol induced BBB breakdown no traces of ICG were found during spectrophotometric analysis of the brain cell suspensions. In ten rats with disrupted BBB there were no significant increases of brain temperature or histological signs of brain damage following 1 h NIR-light exposure after ICG injection. The existing literature concerning the application of ICG in combination with NIR light is reviewed. PMID- 12084563 TI - Non-invasive induction of focal cerebral ischemia in mice by photothrombosis of cortical microvessels: characterization of inflammatory responses. AB - In this study, we adapted the original rat photothrombosis model of Watson et al. (Ann Neurol 17 (1985) 497) for use in mice by refining the application route of the dye, illumination and stereotactic parameters. After intraperitoneal injection of the photosensitive dye Rose bengal, subsequent focal illumination of the brain with a cold light source through the intact skull led to focal cortical infarcts of reproducible size, location and geometry. Cresyl violet histology displayed well-demarcated infarcts that matured with time in a predictable manner. Microglial responses, as assessed by immunocytochemistry, against F4/80 and CD11b antigens were rapid and complete at the infarct site, but delayed and incomplete in degenerating fiber tracts and ipsilateral thalamic nuclei. In contrast to the rat, where the expression of CD4 and CD8 antigens discriminate distinct subpopulations of lesion-associated phagocytes, the expression of both markers was low to absent in the mouse model. In both rats and mice, cerebral photothrombosis shares essential inflammatory responses with focal ischemia induced by middle cerebral artery occlusion. It may provide a useful model to study functional aspects of lesion-associated and remote molecular responses in transgenic mice. PMID- 12084564 TI - Foci identification of spike discharges in the EEGs of sleeping El mice based on the electric field model and wavelet decomposition of multi monopolar derivations. AB - We report on spike discharges in the EEGs of sleeping El mice, which are considered to be homologous to the state seen in human epileptic patients. The discrete wavelet transform (DWT) was used to decompose the EEGs derived from seven electrodes, with the primary spike frequency detected in the 15.6-7.8 Hz detail. The synchronicity of the spikes was evaluated in 98 samples. We succeeded in detecting the foci of the spikes by applying the electric field model, in which the surface potential was expressed as a function of distance between the focus and each electrode based on Gauss' theorem. The foci were obtained in 73 of 91 spikes so evaluated. In 69 of these 73 spikes, the calculated foci occurred within the brain, with none of the foci occurring at identical positions. The distribution occurred in relatively deeper brain regions, suggesting involvement of the paleocortex rather than the hippocampal system. The present study provides reasonable grounds for an explanation of the EEG using the electric field model supported by efficient foci detection of the spikes in a one-to-one correspondence. This paradigm will be applicable to all animal species including humans and will be helpful in the detection of the epileptic focus and in obtaining an insight into the mechanisms of spike genesis. PMID- 12084565 TI - Adaptive time-frequency parametrization in pharmaco EEG. AB - Adaptive time-frequency approximations offer description of the local structures of a signal in terms of their time and frequency coordinates, widths and amplitudes. These parameters can then be used to select and study electroencephalogram (EEG) structures like sleep spindles or slow wave activity (SWA) with high resolution. Such a detailed description of relevant structures improves on the sensitivity of the traditionally used spectral power estimates and opens new possibilities of investigation. These advantages are illustrated using a double-blind test of the influence of zolpidem and midazolam on sleep EEG, and the results are compared with the traditional approach. The observed decrease of frequency of the SWA under the influence of sleep-inducing drugs gives an example of an effect elusive to classical methodology. PMID- 12084566 TI - Axotomy of single fluorescent nerve fibers in developing mammalian spinal cord by photoconversion of diaminobenzidine. AB - A technique has been developed for cutting single nerve fibers in mammalian spinal cord. In the presence of diaminobenzidine (DAB), a laser microbeam was applied to carbocyanine (Dil) stained sensory fibers in cultured spinal cords of the newly born opossum Monodelphis domestica. Digital images of fluorescent fibers were acquired with an intensified video CCD-camera coupled to an image processor. Laser illumination of two spots on a fiber in the presence of 3 mg/ml DAB cut it, so that following DAB wash out, Dil fluorescence did not return after the intermediate segment was bleached. In contrast, when a similar procedure was carried out without DAB, fluorescence of the bleached segment was recovered within minutes in darkness, by dye diffusion from adjacent regions of the uncut fiber. After exposure to DAB, through-conduction of compound action potentials continued in undamaged fibers. The DAB reaction product remained as a dark precipitate, helping to localize the lesion sites. By illuminating a continuous series of spots it was possible to cut whole nerve roots. Fluorescent fibers extended across the cut segment 24 h later. With minor modifications, the procedure described here allows a precise lesioning of single fibers within an intact nervous system. PMID- 12084567 TI - Correlative fluorescence and electron microscopy of biocytin-filled neurons with a preservation of the postsynaptic ultrastructure. AB - Several techniques enable to inject intracellularly neurons with dyes and to use light and electron microscopy to correlate the physiological data with the morphological properties of the neuron. However, the ultrastructure of the neuron is usually obscured by the injected dye thus notably precluding the analysis of the postsynaptic specialisation and that of the other organelles. To overcome this problem, we have developed a technique based on fluorophore- and ultra small gold-conjugated streptavidins. We report, that this method facilitates the identification of intracellular organelles of the biocytin-filled neuron and of postsynaptic densities. This method is valid for the study of early postnatal neurons that are particularly refractory to this type of analysis. The procedure introduced here consists of the following steps: (1) injection of biocytin into the neuron by a patch-clamp pipette, (2) aldehyde fixation, (3) reaction with a fluorophore-conjugated streptavidin, (4) analysis with a fluorescence microscope, (5) formation of avidin-biotin complexes (ABC), (6) reaction with an ultra small gold-conjugated streptavidin, (7) silver enhancement of gold, (8) postfixation with osmium tetroxide and embedding in resin, (9) ultrathin sectioning and analysis with an electron microscope. Using this method, we show that in early postnatal hippocampal neurons, that have been injected with biocytine, it is possible to determine the morphology of the dendritic and axonal trees (including very thin details such as spines and filopodia) and to identify the localisation of the symmetric and asymmetric synapses on dendrites of the injected neuron. PMID- 12084568 TI - Improved electrochemical detection of biogenic amines in Aplysia using base hydrolyzed cellulose-coated carbon fiber microelectrodes. AB - A major challenge with electrochemical techniques in vivo, using carbon-fiber microelectrodes, is to achieve sufficient sensitivity to detect the low concentrations of transmitters released by neurons. In particular, when an electrode is inserted into living tissue, its sensitivity is usually substantially decreased as a result of the degradation of the carbon surface by proteins. Here, we show that this decrease can be significantly attenuated by coating the electrode with cellulose acetate. The cellulose film offers a steric barrier that prevents macromolecules from diffusing to the carbon surface and its porosity can be progressively increased by controlled hydrolysis. We compared different cellulose-coated electrodes, either non-hydrolysed or hydrolyzed, in 0.08 N KOH for 10-30 min. We found that dopamine and serotonin detection was blocked by non-hydrolysed cellulose films, but that hydrolysis restored optimal detection similar to uncoated electrodes. Moreover, cellulose films (hydrolyzed for 20 min) significantly diminished electrode degradation in vivo and allowed reliable detection of fast concentration changes with <0.5 s delay, compared to uncoated electrodes. Finally, the sensitivity to endogenous 5-HT release in Aplysia central nervous system was more than doubled with these electrodes. We conclude that the optimal hydrolysis time of cellulose-coated electrodes is approximately 20 min with our protocol and carbon fiber electrodes prepared with this method offer improved sensitivity for the detection of biogenic amines. PMID- 12084569 TI - Wavelet entropy analysis of event-related potentials indicates modality independent theta dominance. AB - Sensory/cognitive stimulation elicits multiple electroencephalogram (EEG) oscillations that may be partly or fully overlapping over the time axis. To evaluate co-existent multi-frequency oscillations, EEG responses to unimodal (auditory or visual) and bimodal (combined auditory and visual) stimuli were analyzed by applying a new method called wavelet entropy (WE). The method is based on the wavelet transform (WT) and quantifies entropy of short segments of the event-related brain potentials (ERPs). For each modality, a significant transient decrease of WE emerged in the post-stimulus EEG epoch indicating a highly-ordered state in the ERP. WE minimum was always determined by a prominent dominance of theta (4-8 Hz) ERP components over other frequency bands. Event related 'transition to order' was most pronounced and stable at anterior electrodes, and after bimodal stimulation. Being consistently observed across different modalities, a transient theta-dominated state may reflect a processing stage that is obligatory for stimulus evaluation, during which interfering activations from other frequency networks are minimized. PMID- 12084570 TI - A noncoding RNA regulates human protease-activated receptor-1 gene during embryogenesis. AB - Activation of the human protease-activated receptor-1 (PAR-1) by thrombin leads to myriad functions essential for maintaining vascular integrity. Upregulation of PAR-1 expression is considered important in atherosclerosis, angiogenesis and tumor metastasis. In vitro analysis of the human PAR-1 promoter function revealed a positive regulatory element between -4.2 and -3.2 kb of the transcription start site. This element was examined in transgenic mice containing either 4.1 or 2.9 kb of the 5' flanking sequence driving a LacZ reporter gene. Only the 4.1 kb PAR 1 transgene was expressed in vivo and only during embryonic development. The transgene expression was observed only in developing arteries and not in veins. Further examination of this putative regulatory sequence identified a novel noncoding RNA (ncR-uPAR:noncoding RNA upstream of the PAR-1) gene at -3.4 kb. The ncR-uPAR upregulated PAR-1-core promoter-driven luciferase activity and mRNA expression in vitro in a Pol II-dependent manner. This noncoding RNA appears to act in trans, albeit locally at the adjacent PAR-1 promoter. These data suggest that an untranslated RNA plays a role in PAR-1 gene expression during embryonic growth. PMID- 12084571 TI - Molecular cloning and expression of a Toll receptor gene homologue from the silkworm, Bombyx mori. AB - A novel Toll receptor gene was cloned from the bacterial artificial chromosomal (BAC) library of the silkworm, Bombyx mori, using primers for polymerase chain reactions (PCR), synthesized based on an expressed-sequence tag (EST). The open reading frame of this gene was intronless. The deduced amino acid sequence including a putative signal peptide from the nucleotide sequence of the B. mori Toll (BmToll) gene shows 53.7%, 49.0%, 42.3% and 39.2% similarity to those of Toll-7, 18 wheeler (18w), Toll-6 and Toll-8 from Drosophila melanogaster. BmToll, however, does not show marked similarity to Toll, Toll-3, Toll-4 or Toll-5. Phylogenetic insect Toll family analysis shows that BmToll is strongly related to Toll-7 and 18w. An analysis of the tissue-specific expression of the BmToll gene by reverse transcription PCR (RT-PCR) showed that the BmToll gene is constitutively expressed in the fat body but not in the Malpighian tubule, silk gland, midgut or hemocyte. Injecting lipopolysaccharide (LPS) into silkworm hemocoel strongly suppressed BmToll gene expression. The time course showed that BmToll gene expression is totally suppressed within 2 h and gene transcripts appeared again 12 h after LPS injection, suggesting strong downregulation of the BmToll gene expression by LPS. PMID- 12084572 TI - Cloning and molecular characterization of the rat CB2 cannabinoid receptor. AB - The rat peripheral cannabinoid receptor (rCB2) was cloned from a Sprague-Dawley rat spleen cDNA library and when translated, encodes a protein of 410 amino acids. Alignment of rCB2 with mouse (mCB2) and human (hCB2) peripheral cannabinoid receptors reveals a high degree of homology except in the carboxy terminus where rCB2 is 50 and 63 residues longer than hCB2 and mCB2, respectively. PCR screening and sequencing of rat genomic DNA showed that rCB2 is encoded by three exons interrupted by two introns, one of which is polymorphic and contains a 209 base pair B2 (SINE) element. By Northern hybridization and ribonuclease protection assay (RPA), rCB2 mRNA was detected in rat spleen, testis, thymus and lung but not in rat brain, heart, kidney or liver. Like hCB2 and mCB2 receptors, rCB2 activates mitogen-activated protein kinase when it is stably expressed in Chinese Hamster Ovary (CHO) cells. The importance of the carboxy terminus in regulating CB2 receptor desensitization and internalization is well-established. Thus, the profound differences identified in this region of the CB2 receptor between species mandates caution when extrapolating experimental results from non-human models to the effects of chronic CB2 receptor stimulation in humans. PMID- 12084573 TI - Xwnt-2 (Xwnt-2b) is maternally expressed in Xenopus oocytes and embryos. AB - Xwnt-2 (formerly Xwnt-2b) is a member of the Xwnt-8 class of axis-inducing Wnts. Its zygotic expression is at the prosencephalic-mesencephalic border of the early tadpole brain and above the heart primordium [Mech. Dev. 63 (1997) 199]. Here, we report that Xwnt-2 has an earlier, maternal pattern of expression. It is detected in the oocyte, egg and the developing embryo. Studies of the spatial localization of maternal Xwnt-2 show transcripts in both vegetal and animal blastomeres with enrichment in the animal hemisphere. The identification of maternal Xwnt-2 raises questions about possible roles of dorsalizing Xwnts in axial patterning of the Xenopus embryo. PMID- 12084574 TI - Development of multicomponent DNA delivery systems based upon poly(amidoamine) PEG co-polymers. AB - PEGylated polyamidoamine (PAA) polymers were investigated for the production of sterically stabilised DNA delivery systems. Comparison of a PEGylated polymer (NG47) with a non-PEGylated polymer (NG49) showed similar binding of co-polymer to DNA by displacement of ethidium bromide (EB) and DNA melting studies. Gel electrophoresis, turbidimetric analysis and PCS demonstrated differences in the colloidal properties of the complexes, which were attributable to the formation of soluble complexes by the PEGylated co-polymer. However, transmission electron microscopy (TEM) showed that the resulting complexes containing poly(ethylene glycol) (PEG) were not well condensed, susceptible to degradation by nucleases, and thus not suited for in vivo delivery. The poor properties of the PEGylated co polymer were attributed to an excess of PEG. However, polymer blends of NG47 and NG49 at defined ratios of polymer to co-polymer and total repeating units (RUs) to nucleotide, spontaneously formed complexes with a range of desirable properties. These included small size and polydispersity, high particle density, low surface charge and resistance to nuclease degradation. Complexes made with PEGylated polymer alone, and the polymer blends both suffered from a reduced polyfection activity. This was attributed to a low surface charge on the complex, which reduced interactions with the cell membrane and consequent uptake of the particles into the cell. PMID- 12084575 TI - Crooked neck is a component of the human spliceosome and implicated in the splicing process. AB - The Drosophila crooked neck (crn) gene is essential for embryogenesis and has been implicated in cell cycle progression and in pre-mRNA splicing although a direct role in either process has not been established. Here we report isolation of the human crooked neck homolog, HCRN, and provide evidence for its function in splicing. HCRN encodes an unusual protein composed largely of tetratricopeptide repeat (TPR) elements. The crooked neck protein co-localizes with the SR and Sm protein splicing factors in discrete subnuclear domains implicated in snRNP biogenesis. In vitro assembly experiments show that an 83 kDa hcrn isoform is stably recruited to splicing complexes coincident with the addition of the U4/U6.U5 tri-snRNP particle. Crooked neck activity appears essential as extracts depleted of hcrn fail to splice pre-mRNA. These and related data support the view that crooked neck is a phylogenetically conserved pre-mRNA splicing factor. PMID- 12084577 TI - cis-Acting elements within CFTR 5'-flanking DNA are not sufficient to decrease gene expression in response to phorbol ester. AB - The cystic fibrosis transmembrane conductance regulator gene (CFTR) is regulated in a tissue-specific and developmental fashion. Although it has been known for some time that phorbol esters decrease CFTR expression in cell lines that have high CFTR mRNA levels, the cis-acting elements that control this down-regulation remain ill-defined. The role of cis-acting elements within the CFTR minimal promoter in modulating responses to phorbol 12-myristate 13-acetate (PMA) and forskolin was assessed using luciferase reporter gene (luc)-containing plasmids transfected into Calu-3 and HT-29 cells. PMA treatment had no effect on luciferase activity in Calu-3 cells transiently transfected with plasmids containing luc driven by up to 2.3 kb of CFTR 5'-flanking DNA. PMA increased luciferase activity in transfected HT-29 cells. A more extensive region of DNA was evaluated using a yeast artificial chromosome (YAC) containing luc driven by approximately 335 of CFTR 5'-flanking DNA (y5'luc) stably introduced into HT-29 cells. Clonal cell lines containing y5'luc were created and assessed for luciferase activity at baseline and in response to forskolin and PMA. There was a wide range of baseline luciferase activities among the clones (42-1038 units/microg protein) that was not entirely due to the number of luc copies present within the cells. Treatment with both PMA and forskolin led to increased luciferase activity in six randomly selected clonal cell lines. As expected, endogenous CFTR expression increased in response to forskolin and decreased in response to PMA. These studies demonstrate that luc-containing YAC vectors can be used to study CFTR expression in human cells. In addition, these data suggest that important regulatory elements responsible for decreased CFTR expression in response to PMA are not located upstream of CFTR in the approximately 335 kb 5' flanking sequence included in this YAC construct. PMID- 12084576 TI - Hyper-phosphorylation of the adeno-associated virus Rep78 protein inhibits terminal repeat binding and helicase activity. AB - The replication (Rep) proteins of adeno-associated virus (AAV) play prominent roles in regulation of viral DNA replication, RNA transcription, assembly of an infectious virion and establishment of the provirus. We have previously demonstrated that all four Rep proteins are phosphorylated on serine residues [Virology 23 (1997) 332-336]. Reversible phosphorylation may provide a mechanism for regulating Rep protein function. To test this hypothesis, we used the phosphatase inhibitor okadaic acid (OA) to obtain hyper-phosphorylated Rep proteins. OA treatment of AAV- and adenovirus (Ad)-infected cells and baculovirus infected insect cells at a concentration of 100 nM resulted in a significant increase in Rep protein phosphorylation. This concentration suggests that protein phosphatase 2A (PP2A) is one of the enzymes involved in regulation of Rep phosphorylation. The increased phosphorylation occurred primarily on serine residues with a detectable amount of phosphate on threonine. Hyper phosphorylation of Rep78 resulted in reduced binding to the AAV origin of DNA replication. Hyper-phosphorylated Rep78 also had diminished helicase activity. These results suggest that regulated phosphorylation of Rep78 plays a role in controlling Rep functions in the virus replication cycle. PMID- 12084578 TI - Molecular and functional characterization of a second copy of the aflatoxin regulatory gene, aflR-2, from Aspergillus parasiticus. AB - The genes required for the synthesis of aflatoxin (AF) in Aspergillus flavus and Aspergillus parasiticus have been shown to be clustered on a chromosome in these fungi. Transcription of most of these genes is dependent upon the activity of the aflR gene, also present on the gene cluster, which encodes a zinc binuclear cluster DNA-binding protein. While many strains of A. parasiticus have only one copy of aflR (aflR-1), many others contain a second copy of this gene (aflR-2) which resides on a duplicated region of the aflatoxin gene cluster. Targeted disruption of aflR-1 generated a number of non-aflatoxin producing transformants of A. parasiticus SU-1 which still harbored a wild-type aflR-2 gene. Southern and Northern hybridization analyses and ELISA assays demonstrated that aflR-1 had been successfully inactivated in strain AFS10. DNA sequence analysis showed that aflR-2 was capable of encoding a deduced 47 kDa protein. Northern and RT-PCR analysis of RNA from a toxin producing strain indicated that aflR-2 was transcribed at extremely low levels compared to aflR-1. RT-PCR analysis of RNA from AFS10 demonstrated that mRNAs of aflatoxin pathway genes were not processed to their mature forms. Functional analysis of aflr-2 protein in a yeast system showed that it was not activating transcription. PMID- 12084579 TI - Promoter II of the bovine acetyl-coenzyme A carboxylase-alpha-encoding gene is widely expressed and strongly active in different cells. AB - Three promoters express the bovine gene encoding the enzyme acetyl-CoA carboxylase-alpha (ACC alpha) known to be rate-limiting for fatty acid synthesis. Our sequence of promoter II shows that PII is evolutionary conserved, unlike PI or PIII. In vivo expression of PII reveals little tissue-specific restrictions. The proximal 133 bp of this promoter are sufficient for a strong basal expression in different cell lines. PMID- 12084580 TI - Isolation and molecular characterization of the [Fe]-hydrogenase from the unicellular green alga Chlorella fusca. AB - [Fe]-hydrogenases are redoxenzymes that catalyze the reversible reduction of protons to hydrogen. Hydrogenase activity was observed in a culture of the unicellular green alga Chlorella fusca after an anaerobic incubation, but not in the related species Chlorella vulgaris. Specific polymerase chain reaction (PCR) techniques lead to the isolation of the cDNA and the genomic DNA of a special type of [Fe]-hydrogenase in C. fusca. The functional [Fe]-hydrogenase was purified to homogeneity and its N-terminus was sequenced. The polypeptide sequence shows a high degree of identity with the amino acid sequence deduced from the respective cDNA region. Structural and biochemical analyses indicate that ferredoxin is the main physiological electron donor. PMID- 12084581 TI - Cloning and identification of a new member of water channel (AQP10) as an aquaglyceroporin. AB - Recently, a new member of aquaporins was reported as AQP10 [Biochem. Biophys. Res. Commun. 287 (2001) 814], which is incompletely spliced to lose the sixth transmembrane domain and has poor water and no glycerol/urea permeabilities. Independently, we identified a similar clone in human. Our AQP10 consists of 301 amino acids with a highly conserved sixth transmembrane domain. AQP10 has higher identity with aquaglyceroporins (50% with AQP9, 48% with AQP3, 42% with AQP7) and lower identity with other aquaporins (32% with AQP1 and AQP8). AQP10 is expressed only in the small intestine with (approximately 2 kb). RNase protection assay revealed the absence of the unspliced form, supporting the authenticity of our clone. When expressed in Xenopus oocytes, AQP10 stimulated osmotic water permeability sixfold in a mercury-sensitive manner. Glycerol and urea uptakes were also stimulated, while adenine uptake was not. The genome structure of AQP10 is similar to those of other aquaglyceroporins (AQP3, AQP7, AQP9) with six exons. We conclude that AQP10 represents a new member of aquaglyceroporins functionally as well as structurally. PMID- 12084582 TI - Identification of a novel mouse membrane-bound family 1 glycosidase-like protein, which carries an atypical active site structure. AB - We have identified a novel mouse gene klph (Klotho-LPH related protein; where LPH stands for lactase-phlorizin hydrolase) that encodes a novel mammalian family 1 glycosidase-like protein. KLPH was a putative type I membrane protein that consists of N-terminal signal sequence, glycosidase domain, transmembrane region and short cytoplasmic tail. Despite its overall structural similarity to other family 1 glycosidases, the glutamic acid for the acid-base catalyst was not conserved in this protein. klph mRNA was predominantly expressed in the kidney and skin. Epitope-tagged KLPH was localized to the perinuclear tubular network structure of the endoplasmic reticulum in cultured cells. PMID- 12084583 TI - Chloroplast ribosomal S14 protein transcript is edited to create a translation initiation codon in the moss Physcomitrella patens. AB - The rps14 transcript is edited in the moss Physcomitrella patens chloroplast by a C-to-U transition, to create a translation initiation codon, AUG. The efficiency of RNA editing was low, with approximately 20% of rps14 transcripts edited. This suggests that the translation of rps14 mRNA is strictly regulated by RNA editing. This is the first report of RNA editing in P. patens and the creation of a translation initiation codon in rps14 mRNA in chloroplasts. PMID- 12084584 TI - Rationale for on-site cardiac surgery for primary angioplasty: a time for reappraisal. AB - Since the early 1990s, with significant improvement in the procedural success of percutaneous coronary interventions (PCIs), there has been a concomitant reduction in the need for emergency coronary artery bypass graft surgery. This review article focuses on the need for on-site cardiac surgery in patients with acute myocardial infarction undergoing primary angioplasty at centers without on site cardiac surgical backup. It gives an overview of the need for emergency bypass surgery in both the large trial setting and the community hospital setting. Special consideration is also given to the risks and benefits of primary angioplasty compared with thrombolytic therapy, transfer to an institution with an on-site cardiac surgical facility compared with primary PCI, the frequency and indications for emergency cardiac surgery related and unrelated to primary angioplasty and the requirements for primary angioplasty that must be met in hospitals without the capability of on-site cardiac surgery. PMID- 12084585 TI - The incidence of congenital heart disease. AB - This study was designed to determine the reasons for the variability of the incidence of congenital heart disease (CHD), estimate its true value and provide data about the incidence of specific major forms of CHD. The incidence of CHD in different studies varies from about 4/1,000 to 50/1,000 live births. The relative frequency of different major forms of CHD also differs greatly from study to study. In addition, another 20/1,000 live births have bicuspid aortic valves, isolated anomalous lobar pulmonary veins or a silent patent ductus arteriosus. The incidences reported in 62 studies published after 1955 were examined. Attention was paid to the ways in which the studies were conducted, with special reference to the increased use of echocardiography in the neonatal nursery. The total incidence of CHD was related to the relative frequency of ventricular septal defects (VSDs), the most common type of CHD. The incidences of individual major forms of CHD were determined from 44 studies. The incidence of CHD depends primarily on the number of small VSDs included in the series, and this number in turn depends upon how early the diagnosis is made. If major forms of CHD are stratified into trivial, moderate and severe categories, the variation in incidence depends mainly on the number of trivial lesions included. The incidence of moderate and severe forms of CHD is about 6/1,000 live births (19/1,000 live births if the potentially serious bicuspid aortic valve is included), and of all forms increases to 75/1,000 live births if tiny muscular VSDs present at birth and other trivial lesions are included. Given the causes of variation, there is no evidence for differences in incidence in different countries or times. PMID- 12084586 TI - The regulation and measurement of plasma volume in heart failure. AB - Plasma volume, the intravascular portion of the extracellular fluid volume, can be measured using standard dilution techniques with radiolabeled tracer molecules. In healthy persons, plasma volume remains relatively constant as a result of tight regulation by the complex interaction between neurohormonal systems involved in sodium and water homeostasis. Although chronic heart failure (CHF) is characterized by activation of many of these neurohormonal systems, few studies have evaluated plasma volume in this condition under treatment. Untreated edematous decompensated heart failure (HF) is associated with a significant expansion of plasma volume. Patients with stable CHF, receiving conventional therapy, appear to have a contracted plasma volume, a concept that is in contrast to the widely held belief that CHF is associated with long-term hypervolemia. It is likely that significant changes in plasma volume occur during intensification of medical therapy or during transition from the edematous to the stable state. Clinical assessment of plasma volume may be of particular value during treatment in patients with decompensated HF, in whom the plasma volume is contracted despite an increase in total extracellular fluid volume. Under these circumstances, treatment with inotropes or renal vasodilators may be more appropriate than intravenous diuretics alone. Further studies evaluating plasma volume in HF may help to improve our understanding of the pathophysiologic mechanisms occurring in the development and progression of this complex condition. PMID- 12084587 TI - Biology or bias: practice patterns and long-term outcomes for men and women with acute myocardial infarction. AB - OBJECTIVES: The goal of our study was to examine how age and gender affect the use of coronary angiography and the intensity of cardiac follow-up care within the first year after acute myocardial infarction (AMI). Another objective was to evaluate the association of age, gender and treatment intensity with five-year survival after AMI. BACKGROUND: Utilization rates of specialized cardiac services inversely correlate with age. Gender-specific practice patterns may also vary with age in a manner similar to known age-gender survival differences after AMI. METHODS: Using linked population-based administrative data, we examined the association of age and gender with treatment intensity and long-term survival among 25,697 patients hospitalized with AMI in Ontario between April 1, 1992, and December 31, 1993. A Cox proportional hazards model was used to adjust for socioeconomic status, illness severity, attending physician specialty and admitting hospital characteristics. RESULTS: After adjusting for baseline differences, the relative rates of angiography and follow-up specialist care for women relative to men, respectively, fell 17.5% (95% confidence interval [CI], 13.6 to 21.3, p < 0.001) and 10.2% (95% CI, 7.1 to 13.2, p < 0.001) for every 10 year increase in age. Conversely, long-term AMI survival rates in women relative to men improved with increasing age, such that the relative survival in women rose 14.2% (95% CI, 10.1 to 17.5, p < 0.001) for every 10-year age increase. CONCLUSIONS: Gender differences in the intensity of invasive testing and follow up care are strongly age-specific. While care becomes progressively less aggressive among older women relative to men, survival advantages track in the opposite direction, with older women clearly favored. These findings suggest that biology is likely to remain the main determinant of long-term survival after AMI for women. PMID- 12084588 TI - Systemic inflammation in unstable angina is the result of myocardial necrosis. AB - OBJECTIVES: We investigated whether the source of the acute phase response in unstable angina (UA) lay within the culprit coronary plaque or distal myocardium. BACKGROUND: An inflammatory response is an important component of the acute coronary syndromes. However, its origin and mechanism remain unclear. METHODS: In 94 stable patients undergoing coronary angiography, the relationship between systemic levels of tumor necrosis factor (TNF)-alpha, interleukin-6 (IL-6) and C reactive protein (CRP) and extent of atherosclerosis was studied. The temporal relationship between these markers and troponin T (TnT) was determined in 91 patients with UA. Cytokine levels were measured in the aortic root and coronary sinus of 36 unstable patients. RESULTS: There was no relationship found between stable coronary atherosclerosis and inflammatory marker levels. Compared with this group, admission levels of IL-6 (3.6 +/- 0.3 ng/ml vs. 10.7 +/- 1.7 ng/ml, p < 0.05) and CRP (2.3 +/- 0.1 mg/l vs. 4.6 +/- 0.6 mg/l, p < 0.05) were elevated in patients with UA. In this group, IL-6 and CRP remained elevated in those who subsequently experienced major adverse cardiac events. This inflammatory response occurred in parallel to the appearance of TnT. Both TNF-alpha (19.2 +/- 3.4 ng/ml vs. 17.1 +/- 3.3 ng/ml, p < 0.001) and IL-6 (10.3 +/- 1.4 ng/ml vs. 7.7 +/- 1.1 ng/ml, p < 0.01) were elevated in the coronary sinus compared with aortic root in patients with UA. This was principally observed in those who were TnT positive. There was no cytokine gradient across the culprit plaque. CONCLUSIONS: There is an intracardiac inflammatory response in UA that appears to be the result of low grade myocardial necrosis. The ruptured plaque does not appear to contribute to the acute phase response. PMID- 12084589 TI - Early angioplasty in acute coronary syndromes without persistent ST-segment elevation improves outcome but increases the need for six-month repeat revascularization: an analysis of the PURSUIT Trial. Platelet glycoprotein IIB/IIIA in Unstable angina: Receptor Suppression Using Integrilin Therapy. AB - OBJECTIVES: We explored the effect of timing of percutaneous coronary intervention (PCI) in acute coronary syndromes (ACS) without persistent ST segment elevation on the need for repeat revascularization, and we related this effect to other events. BACKGROUND: Percutaneous coronary intervention is widely used to treat ACS without persistent ST-segment elevation. Moreover, restenosis and subsequent revascularization after PCI are more frequent in ACS than in stable angina. The optimal timing of PCI in ACS without persistent ST-segment elevation is unknown. METHODS: In the Platelet glycoprotein IIB/IIIA in Unstable angina: Receptor Suppression Using Integrilin Therapy (PURSUIT) database, patients were stratified by the time of PCI. In the PURSUIT trial, 9,461 patients received a platelet glycoprotein IIb/IIIa inhibitor, eptifibatide or placebo for 72 h. The investigators decided on other treatments. RESULTS: A total of 2,430 patients underwent PCI within 30 days. Repeat revascularization (during 165 days) was notably higher for PCI within 24 h of enrollment (n = 620 [19%]) than for PCI at 24 to 72 h (n = 624 [16.7%]), 3 to 7 days (n = 614 [13.2%]), or 8 to 30 days (n = 561 [7.7%]; p < 0.001), regardless of eptifibatide use. This gradual reduction in the revascularization rate for later PCI was also observed after multivariate analysis correcting for baseline characteristics and with time as a continuous variable. CONCLUSIONS: Percutaneous coronary intervention within 24 is associated with improved outcome (other analysis) but more repeat revascularization. Prospective analyses are needed to test the hypothesis that rapid PCI in ACS with a platelet glycoprotein IIb/IIIa receptor antagonist reduces myocardial infarction (and possibly death) and is therefore most suited for patients at highest risk of infarction, despite a higher need for repeat revascularization. PMID- 12084590 TI - The effect of intracoronary radiation for the treatment of recurrent in-stent restenosis in patients with diabetes mellitus. AB - OBJECTIVES: The purpose of this study was to examine the effect of intracoronary radiation therapy (IRT) in diabetic patients with in-stent restenosis (ISR). BACKGROUND: Diabetic patients are at an increased risk for restenosis, repeat revascularization procedures and late mortality after percutaneous coronary interventions and stenting. Intracoronary radiation therapy, utilizing both gamma and beta-emitters, has been shown to reduce the rate of ISR. METHODS: The study group consisted of 749 consecutive patients with ISR who were treated with either IRT or placebo in randomized trials and registries at our center. Diabetic patients (252 radiation and 51 placebo) were compared with nondiabetic patients (371 radiation and 75 placebo). RESULTS: In-hospital outcomes were similar between diabetic and nondiabetic patients treated with and without radiation. At six-month clinical and angiographic follow-up, there was a significant reduction in the binary restenosis (63.8% vs. 15.7%, p < 0.0001), target lesion revascularization (66.7% vs. 17.6%, p < 0.0001) and target vessel revascularization (TVR) (70.6% vs. 22.9%, p < 0.0001) rates in diabetic patients treated with radiation compared to placebo. Comparisons between the placebo arms detected a trend towards higher restenosis (63.8% vs. 48.4% p = 0.13) and TVR (70.6% vs. 56.0%, p = 0.14) in diabetic versus nondiabetic patients. In contrast, diabetic and nondiabetic patients treated with IRT experienced similar restenosis (15.6% vs. 10.7% p = 0.33) and TVR (22.9% vs. 28.2% p = 0.41) rates. CONCLUSIONS: In diabetic patients with ISR, intracoronary radiation significantly reduced the recurrence of ISR compared to placebo. Additionally, similar rates of restenosis and revascularization procedures were achieved in irradiated diabetic and nondiabetic patients. In view of these results, IRT should be considered as a valuable therapeutic alternative in all diabetic patients with ISR. PMID- 12084591 TI - Dose heterogeneity may not affect the neointimal proliferation after gamma radiation for in-stent restenosis: a volumetric intravascular ultrasound dosimetric study. AB - OBJECTIVES: The goal of this study was to use serial (postirradiation and follow up) volumetric intravascular ultrasound (IVUS): 1) to evaluate the actual distribution of gamma radiation in human in-stent restenosis (ISR) lesions, and 2) to analyze the relationship between neointimal regrowth and the delivered radiation dose. BACKGROUND: The relationship between the neointimal regrowth and delivered dose during the treatment of ISR remains unknown. METHODS: We analyzed 20 actively (gamma emitter) treated, native artery ISR patients from the Washington Radiation for In-Stent restenosis Trial (WRIST) that met the following criteria: on both postirradiation and six-month follow-up IVUS imaging, > or =80% of the external elastic membrane circumference could be identified throughout the treated length including the lesion and proximal and distal reference segments. Intravascular ultrasound images were digitized every 1 mm. Proximal and distal reference and stented segment luminal and adventitial contours were imported and reconstructed. The source was placed circumferentially at the site of the IVUS catheter and longitudinally according to the relationship between the radioactive seeds and stent edges. Using Monte Carlo simulations, dose volume histograms for the adventitia and intima were calculated. The relationship between the neointimal regrowth and calculated doses were evaluated. RESULTS: There was large dose heterogeneity at both the intimal and adventitial levels. Most of the sites (93%) received >4 Gy at the adventitia, and all of the sites received >4 Gy at the intima. There was no relationship between neointimal regrowth and radiation dose. CONCLUSIONS: Although there may be large dose heterogeneity, gamma irradiation (using a fixed dose prescription) appears to deliver a sufficient dose to prevent neointimal regrowth. PMID- 12084592 TI - Nitroglycerin upregulates matrix metalloproteinase expression by human macrophages. AB - OBJECTIVES: This study aimed to determine whether nitroglycerin (NTG) treatment affects matrix metalloproteinase (MMP) gene expression and activities in human macrophages. BACKGROUND: Nitroglycerin is one of the most frequently used therapeutic agents for the symptomatic relief of stable or unstable coronary artery disease; however, its effects on vascular biology are poorly characterized. Despite its powerful vasodilator activity, NTG has not been shown to improve outcomes in coronary disease. We now describe evidence that NTG has potentially pro-inflammatory effects in human monocyte-derived macrophages (MDMs). METHODS: Human monocytes were isolated from whole blood by elutriation and allowed to differentiate into macrophages over eight to 10 days. The MDMs were then treated for 4 or 24 h with control media, pharmacologically relevant doses of NTG or other nitric oxide donors. Matrix metalloproteinase activity was measured by zymography, protein levels measured by enzyme-linked immunosorbent assay and messenger ribonucleic acid (mRNA) levels were quantified by competitive reverse transcription-polymerase chain reaction. RESULTS: The major MMP expressed by MDMs was MMP-9. Nitroglycerin treatment stimulated a dose-dependent increase in MMP-9 mRNA levels (NTG 200 pmol: 193 +/- 6% and NTG 2,000 pmol: 372 +/- 9% compared to controls, p < 0.005) and MMP-9 activity (NTG 200: 142 +/- 5.5% and NTG 2,000: 167 +/- 11% compared to controls, p < 0.005). Nitroglycerin 2,000 pmol also increased MMP-2 and MMP-7 mRNA levels to 187 +/- 8% and 183 +/- 21% of control values, respectively (p < 0.05). Furthermore, tissue inhibitor of metalloproteinase (TIMP)-1 (the major tissue inhibitor of MMPs) mRNA and protein levels were decreased in NTG 2,000 pmol-treated MDMs compared with control cells (mRNA: 67 +/- 7%, p < 0.005; protein: 45 +/- 5%, p < 0.005). CONCLUSIONS: Nitroglycerin in pharmacologically relevant concentrations activates MMP but represses TIMP expression in human macrophages. The subsequent imbalance in MMP/TIMP expression associated with NTG treatment could promote matrix degradation, with potentially adverse effects on plaque stability. PMID- 12084593 TI - Atorvastatin decreases vascular endothelial growth factor in patients with coronary artery disease. AB - OBJECTIVES: The aim of this study was to test a possible influence of atorvastatin on the production of vascular endothelial growth factor (VEGF) in patients with coronary artery disease (CAD) and in vitro. BACKGROUND: Vascular endothelial growth factor is suggested to be involved in the growth of atherosclerotic plaque by inducing its neovascularization. Hepatic hydroxymethyl glutaryl-coenzyme A reductase inhibitors (statins) are known to have atheroprotective effects beyond lipid lowering. METHODS: Blood was collected from 14 male hypercholesterolemic patients with angiographically confirmed CAD at baseline and after two months of atorvastatin therapy (20 mg/d) and from eight male control patients. In an ex vivo assay, human coronary artery smooth muscle cells (HCASMC) were incubated with the patient plasma collected before and after atorvastatin therapy. To test the direct effect of atorvastatin on VEGF synthesis in vitro, HCASMC were treated with atorvastatin (1, 3 and 10 microM). The VEGF concentration was measured by enzyme-linked immunosorbent assay. RESULTS: Atorvastatin therapy reduced VEGF plasma levels in CAD patients (from 31.1 +/- 6.1 to 19.0 +/- 3.6 pg/ml; p < 0.05). The VEGF plasma concentration tended to be higher in CAD patients before treatment compared to control patients (31.1 +/- 6.1 vs. 23.4 +/- 3.6 pg/ml; p = NS). Plasma collected before therapy induced significantly more VEGF in HCASMC compared to the plasma collected after treatment and compared to control cells. In vitro, atorvastatin decreased both the basal and the interleukin-1beta-induced VEGF release in HCASMC. CONCLUSIONS: These data suggest that atorvastatin may lower the plasma level of VEGF in CAD patients, which could represent a novel beneficial effect of this and perhaps other statins. PMID- 12084594 TI - Biphasic versus monophasic shock waveform for conversion of atrial fibrillation: the results of an international randomized, double-blind multicenter trial. AB - OBJECTIVES: This study compared a biphasic waveform with a conventional monophasic waveform for cardioversion of atrial fibrillation (AF). BACKGROUND: Biphasic shock waveforms have been demonstrated to be superior to monophasic shocks for termination of ventricular fibrillation, but data regarding biphasic shocks for conversion of AF are still emerging. METHODS: In an international, multicenter, randomized, double-blind clinical trial, we compared the effectiveness of damped sine wave monophasic versus impedance-compensated truncated exponential biphasic shocks for the cardioversion of AF. Patients received up to five shocks, as necessary for conversion: 100 J, 150 J, 200 J, a fourth shock at maximum output for the initial waveform (200 J biphasic, 360 J monophasic) and a final cross-over shock at maximum output of the alternate waveform. RESULTS: Analysis included 107 monophasic and 96 biphasic patients. The success rate was higher for biphasic than for monophasic shocks at each of the three shared energy levels (100 J: 60% vs. 22%, p < 0.0001; 150 J: 77% vs. 44%, p < 0.0001; 200 J: 90% vs. 53%, p < 0.0001). Through four shocks, at a maximum of 200 J, biphasic performance was similar to monophasic performance at 360 J (91% vs. 85%, p = 0.29). Biphasic patients required fewer shocks (1.7 +/- 1.0 vs. 2.8 +/- 1.2, p < 0.0001) and lower total energy delivered (217 +/- 176 J vs. 548 +/- 331 J, p < 0.0001). The biphasic shock waveform was also associated with a lower frequency of dermal injury (17% vs. 41%, p < 0.0001). CONCLUSIONS: For the cardioversion of AF, a biphasic shock waveform has greater efficacy, requires fewer shocks and lower delivered energy, and results in less dermal injury than a monophasic shock waveform. PMID- 12084595 TI - Intracardiac echocardiography-guided, anatomically based radiofrequency ablation of focal atrial fibrillation originating from pulmonary veins. AB - OBJECTIVES: In patients with a pulmonary vein (PV) source for atrial fibrillation (AF), we sought the use of intracardiac echocardiography (ICE) to evaluate PV anatomy, guide radiofrequency (RF) ablation and monitor for acute stenosis during ablation. BACKGROUND: A focal source for AF may be found in the proximal component of the PVs and can be effectively treated by ablative techniques. However, the procedure may be challenging due to the complex anatomy of the left atrium and PVs, uncertain catheter positioning within the PVs and difficulties in mapping atrial extrasystoles, which may be rare or repeatedly induce AF and require cardioversion. METHODS: Sixty-four patients were referred for RF ablation of a focal source of AF, and 56 were identified as having AF triggers in > or =1 PV. Using ICE guidance, RF lesions were applied around the circumference of the vein near the os until there was electrical isolation. RESULTS: Lesions were placed in 82 veins (36 right superior PV, 33 left superior PV, 9 left inferior PV, 4 right inferior PV); 24 +/- 12 lesions per vein were necessary to create electrical isolation with a fluoroscopic time of 11 +/- 4 min and a mean of 22% reduction in luminal area. After a follow-up of 13 +/- 7 months, 66% of patients remained free of AF, and another 13% responded better to medications. CONCLUSIONS: We describe an anatomic approach to PV electrical isolation in which ICE is used to define the anatomy, guide RF ablation and monitor for acute PV changes. PMID- 12084596 TI - Lesional tachycardias related to mitral valve surgery. AB - OBJECTIVES: The purpose of this study was to define the anatomic distribution of electrically abnormal atrial tissue and mechanisms of atrial tachycardia (AT) after mitral valve (MV) surgery. BACKGROUND: Atrial tachycardia is a well recognized long-term complication of MV surgery. Because atrial incisions from repair of congenital heart defects provide a substrate for re-entrant arrhythmias in the late postoperative setting, we hypothesized that atriotomies or cannulation sites during MV surgery also contributed to postoperative arrhythmias. METHODS: In 10 patients with prior MV surgery, electroanatomic maps were constructed of 11 tachycardias (6 right atrium [RA], 4 left atrium [LA] and 1 biatrial). Activation and voltage maps were used to identify areas of low voltage, double potentials and conduction block. RESULTS: Lesions were present in the lateral wall of the RA (six of seven maps) and in the LA along the septum adjacent to the right pulmonary veins (four of five maps). In 8 of 10 patients, these findings corresponded to atrial incisions or cannulation sites. Arrhythmia mechanisms were identified for 9 of 11 tachycardias. A macro-re-entrant circuit was mapped in six cases, three involving lesions in the lateral wall of the RA and three involving the LA septum and right pulmonary veins. In three of these cases figure-of-eight re-entry was demonstrated, and in the other three a single macro-re-entrant circuit was observed. In three other cases, a focal origin was identified adjacent to abnormal tissue in the RA (two cases) or within a pulmonary vein (one case). CONCLUSIONS: Surgical incisions for MV surgery provide a substrate for atrial arrhythmias. Both macro-re-entrant and focal mechanisms contribute to AT after MV surgery. PMID- 12084597 TI - Differential effects of beta-blockade on dispersion of repolarization in the absence and presence of sympathetic stimulation between the LQT1 and LQT2 forms of congenital long QT syndrome. AB - OBJECTIVES: This study compared the effects of beta-blockade on transmural and spatial dispersion of repolarization (TDR and SDR, respectively) between the LQT1 and LQT2 forms of congenital long QT syndrome (LQTS). BACKGROUND: The LQT1 form is more sensitive to sympathetic stimulation and more responsive to beta-blockers than either the LQT2 or LQT3 forms. METHODS: Eighty-seven-lead, body-surface electrocardiograms (ECGs) were recorded before and after epinephrine infusion (0.1 microg/kg body weight per min) in the absence and presence of oral propranolol (0.5-2.0 mg/kg per day) in 11 LQT1 patients and 11 LQT2 patients. The Q-T(end) interval, the Q-T(peak) interval and the interval between T(peak) and T(end) (T(p-e)), representing TDR, were measured and averaged from 87-lead ECGs and corrected by Bazett's method (corrected Q-T(end) interval [cQT(e)], corrected Q-T(peak) interval [cQT(p)] and corrected interval between T(peak) and T(end) [cT(p-e)]). The dispersion of cQT(e) (cQT(e)-D) was obtained among 87 leads and was defined as the interval between the maximum and minimum values of cQT(e). RESULTS: Propranolol in the absence of epinephrine significantly prolonged the mean cQT(p) value but not the mean cQT(e) value, thus decreasing the mean cT(p-e) value in both LQT1 and LQT2 patients; the differences with propranolol were significantly larger in LQT1 than in LQT2 (p < 0.05). The maximum cQT(e), minimum cQT(e) and cQT(e)-D were not changed with propranolol. Propranolol completely suppressed the influence of epinephrine in prolonging the mean cQT(e), maximum cQT(e) and minimum cQT(e) values, as well as increasing the mean cT(p-e) and cQT(e)-D values in both groups. CONCLUSIONS: Beta-blockade under normal sympathetic tone produces a greater decrease in TDR in the LQT1 form than in the LQT2 form, explaining the superior effectiveness of beta-blockers in LQT1 versus LQT2. Beta-blockers also suppress the influence of sympathetic stimulation in increasing TDR and SDR equally in LQT1 and LQT2 syndrome. PMID- 12084598 TI - Epicardial electrogram of the right ventricular outflow tract in patients with the Brugada syndrome: using the epicardial lead. AB - OBJECTIVES: We tried to record an epicardial electrogram directly, and we examined local electrograms before and after administration of a class IC anti arrhythmic drug in patients with the Brugada syndrome. BACKGROUND: Electrical heterogeneity of the epicardium in the right ventricular outflow tract (RVOT) has been thought to be related to the Brugada syndrome. However, an epicardial abnormality has not been demonstrated in patients with the Brugada syndrome. METHODS: In five patients with a Brugada-type electrocardiogram (ECG), local unipolar electrograms were recorded at the epicardium and endocardium of the RVOT. To record the epicardial electrogram directly, we introduced an electrical guidewire into the conus branch (CB) of the right coronary artery. The duration of the local electrogram after termination of the QRS complex (DP) was measured before and after class IC anti-arrhythmic drug administration. The signal averaged electrocardiogram (SAECG) was also obtained in all patients. RESULTS: A definite DP was observed at the epicardium, but not at the endocardium. After administration of a class IC anti-arrhythmic drug, the DP at the epicardium was prolonged from 38 +/- 10 ms to 67 +/- 24 ms. The late potential corresponding to the DP at the epicardium was observed in all patients on the SAECG. CONCLUSIONS: An epicardial electrogram can be recorded from the CB. Recording from the CB enables identification of an epicardial abnormality in patients with the Brugada syndrome. These abnormal electrograms may be related to a myocardial abnormality in the epicardium of patients with the Brugada syndrome. PMID- 12084599 TI - Late potentials and the Brugada syndrome. PMID- 12084600 TI - Clinical correlates of PR-segment depression in asymptomatic patients with pericardial effusion. AB - OBJECTIVES: The purpose of this study was to determine the clinical correlates of PR-segment depression among consecutive asymptomatic patients with pericardial effusion (PE) detected by routine echocardiography. BACKGROUND: Pericardial effusion is a relatively common finding in clinical practice, but not many studies have evaluated electrocardiographic (ECG) changes associated with the occurrence of PE. METHODS: Among 4,061 consecutive patients referred to our echocardiography laboratory, 176 asymptomatic patients had PE correlated with their clinical history and ECG findings. RESULTS: PR-segment depression was detected in 40 patients (23%). There were no significant differences in age, gender distribution or heart rate between patients with and without PR-segment depression. Fifteen post-pericardiotomy patients (33%), 19 patients (40%) with malignant disease and 6 patients (46%) with connective tissue disease had PR segment depression, whereas no patient with heart disease (dilated cardiomyopathy, hypertensive heart disease, old myocardial infarction, valvular heart disease), renal disease or hypothyroidism had PR-segment depression, nor widespread ST-segment elevation. Among 40 patients with PR-segment depression, 8 had ST-segment elevation in the leads of epicardial derivation, 8 had upright T waves, 20 had low to inverted T waves with an isoelectric ST-segment and 4 had ST T-wave changes due to bundle branch block. CONCLUSIONS: PR-segment depression was a relatively common ECG sign associated with clinically silent PE, and it was an ECG indicator of inflammatory pericardial involvement. PMID- 12084601 TI - Prognostic significance of left ventricular diastolic dysfunction in essential hypertension. AB - OBJECTIVES: We sought to assess the prognostic value of alterations in left ventricular (LV) diastolic function in patients with essential hypertension. BACKGROUND: Alterations in LV diastolic function are frequent in patients with hypertension, even in the absence of LV hypertrophy, but their prognostic significance has never been investigated. METHODS: In the setting of the Progetto Ipertensione Umbria Monitoraggio Ambulatoriale (PIUMA) study, we followed, for up to 11 years (mean: 4.4 years), 1,839 Caucasian hypertensive patients (50 +/- 12 years, 53% men, blood pressure (BP) 156/98 mm Hg) without previous cardiovascular events, who underwent Doppler echocardiography and 24-h BP monitoring before therapy. The early/atrial (E/A) mitral flow velocity ratio was calculated and corrected for age and heart rate (HR). RESULTS: During follow-up, there were 164 major cardiovascular events (2.04 per 100 patient-years). The incidence of cardiovascular events was 2.47 and 1.65 per 100 patient-years in patients with an age- and HR-adjusted E/A ratio below (n = 919) and above (n = 920) the median value, respectively (p < 0.005 by the log-rank test). In Cox analysis, controlling for age, gender, diabetes, cholesterol, smoking, LV mass and 24-h systolic BP (all p < 0.05), a low age- and HR-adjusted E/A ratio conferred an increased risk of cardiovascular events (odds ratio 1.57, 95% confidence interval [CI] 1.11 to 2.18, p < 0.01). A 21% excess risk was found for each 0.3 decrease of the adjusted E/A ratio (95% CI from +2% to +43%; p = 0.03). CONCLUSIONS: Impaired LV early diastolic relaxation, detected by pulsed Doppler echocardiography, identifies hypertensive patients at increased cardiovascular risk. Such association is independent of LV mass and ambulatory BP. PMID- 12084602 TI - Noninvasive assessment of microvascular function in arterial hypertension by transthoracic Doppler harmonic echocardiography. AB - OBJECTIVES: The present study sought to investigate the use of transthoracic Doppler harmonic echocardiography (TTDHE) to evaluate changes in coronary flow dynamics due to microvascular dysfunction. BACKGROUND: Coronary flow velocity reserve (CFVR) measurements by TTDHE are useful for assessing epicardial coronary artery stenoses. It remains unclear, however, if microvascular disease can be detected. METHODS: In 54 patients with chest pain, intracoronary Doppler (ICD) and TTDHE were used to measure average peak velocity at baseline and hyperemia. Significant coronary lesions had been ruled out by both angiography and intravascular ultrasound. Comparative measurements were performed in the distal left anterior descending coronary artery after intracoronary and intravenous administration of adenosine, and CFVR was calculated. Hypertensive patients (n = 25) were studied and compared to a control group (26 normotensive individuals). RESULTS: Three patients (5%) had to be excluded because of insufficient image quality or side effects. In both groups, TTDHE-derived CFVR data correlated closely with ICD measurements (group 1: y = 0.67x + 0.076, standard error of estimate [SEE] = 0.25, r = 0.87, p < 0.001; group 2: y = 0.64x + 1.11, SEE = 0.26, r = 0.87, p < 0.001). CFVR was lower in hypertensives than in normotensive controls (2.44 +/- 0.49 vs. 3.33 +/- 0.40, p < 0.001, cut point = 2.84). CONCLUSIONS: The newly described echocardiographic method is suitable for assessing microvascular dysfunction noninvasively and corresponds well to invasive measurements. PMID- 12084603 TI - Transcatheter closure of patent foramen ovale in patients with cerebral ischemia. AB - OBJECTIVES: The present study was conducted to determine the safety of the transcatheter closure of a patent foramen ovale (PFO) in patients with cryptogenic cerebral ischemia and the midterm follow-up of recurrent thromboembolic events after interventional PFO closure. BACKGROUND: Current therapeutic options for stroke prevention in patients with PFO and a history of thromboembolic events include chronic antithrombotics and more invasive treatments such as surgical closure or minor invasive transcatheter permanent closure of the PFO. Promising preliminary and pilot data with the Amplatzer Septal Occluder or the PFO-Star Occluder have been reported. Systematic and long term data are still missing. METHODS: A total of 276 consecutive patients with a PFO and a history of at least one thromboembolic event were recruited in four medical centers and underwent percutaneous PFO closure with the PFO-Star device. Follow-up data were analyzed over an average of 15.1 months, equivalent to 345 patient-years. RESULTS: The implantation was successful in all 276 patients. Peri interventional reversible complications included transient ST-segment elevations (1.8%) and transient ischemic attack (TIA) (0.8%). Two devices have been removed surgically. During follow-up the annual recurrence rate of thromboembolic events was 1.7% for TIA, 0% for stroke and 0% for peripheral emboli. CONCLUSIONS: Interventional PFO closure with the PFO-Star device appears to be a reliable and promising technique resulting in a low recurrence rate of thromboembolic events, especially stroke in patients with a history of cryptogenic ischemia presumably due to paradoxical embolization. To our knowledge, this is the largest coherent and prospective study for interventional PFO closure. PMID- 12084604 TI - Long-term clinical effect of hemodynamically optimized cardiac resynchronization therapy in patients with heart failure and ventricular conduction delay. AB - OBJECTIVES: We sought to compare the short- and long-term clinical effects of atrial synchronous pre-excitation of one (univentricular) or both ventricles (biventricular), that provide cardiac resynchronization therapy (CRT). BACKGROUND: In patients with heart failure (HF) who have a ventricular conduction delay, CRT improves systolic hemodynamic function. The clinical benefit of CRT is still being investigated. METHODS: Forty-one patients were randomized to four weeks of first treatment with biventricular or univentricular stimulation, followed by four weeks without treatment, and then four weeks of a second treatment with the opposite stimulation. The best CRT stimulation was continued for nine months. Cardiac resynchronization therapy was optimized by hemodynamic testing at implantation. The primary end points were exercise capacity measures. Data were analyzed by two-way repeated-measures analysis of variance. RESULTS: The left ventricle was selected for univentricular pacing in 36 patients. The clinical effects of univentricular and biventricular CRT were not significantly different. The results of each method were pooled to assess sequential treatment effects. Oxygen uptake during bicycle exercise increased from 9.48 to 10.4 ml/kg/min at the anaerobic threshold (p = 0.03) and from 12.5 to 14.3 ml/kg/min at peak exercise (p < 0.001) with the first treatment, and from 10.0 to 10.7 ml/kg/min at the anaerobic threshold (p = 0.2) and from 13.4 to 15.2 ml/kg/min at peak exercise (p = 0.002) with the second treatment. The 6-min walk distance increased from 342 m at baseline to 386 m after the first treatment (p < 0.001) and to 416 m after the second treatment (p = 0.03). All improvements persisted after 12 months of therapy. CONCLUSIONS: Cardiac resynchronization therapy produces a long-term improvement in the clinical symptoms of patients with HF who have a ventricular conduction delay. The differences between optimized biventricular and univentricular therapy appear to be small for short-term treatment. PMID- 12084605 TI - Vasopeptidase inhibition with omapatrilat in chronic heart failure: acute and long-term hemodynamic and neurohumoral effects. AB - OBJECTIVES: We investigated the acute and long-term hemodynamic and neurohumoral effects of the vasopeptidase inhibitor omapatrilat in human heart failure. BACKGROUND: Angiotensin-converting enzyme (ACE) inhibition constitutes a major advance in the treatment of chronic heart failure (CHF). Simultaneous inhibition of both neutral endopeptidase and ACE with omapatrilat may represent a new treatment strategy in CHF. METHODS: Three hundred and sixty-nine patients with symptomatic heart failure were randomized to double-blind treatment with omapatrilat (first 190 patients: 2.5 mg, 5 mg or 10 mg; last 179 patients: 2.5 mg, 20 mg or 40 mg once daily) for 12 weeks. RESULTS: Acutely, the 10 mg, 20 mg and 40 mg doses of omapatrilat produced greater reductions in pulmonary capillary wedge pressure (PCWP), systolic blood pressure (SBP) and systemic vascular resistance compared with 2.5 mg. Higher doses were associated with greater increases in vasodilator and natriuretic peptides, in addition to ACE inhibition. After 12 weeks, omapatrilat 20 mg and 40 mg showed greater falls from baseline in PCWP (40 mg: 0 h to 12 h average change -7.3 +/- 0.8 mm Hg) and SBP (40 mg: -11.7 +/- 1.7 mm Hg) than 2.5 mg (both p < 0.01 vs. 2.5 mg). The incidence of adverse experiences and patient withdrawal were similar in all groups. CONCLUSIONS: In CHF, the acute hemodynamic benefit seen with higher doses of omapatrilat was associated with increases in plasma vasodilator and natriuretic peptide levels in addition to ACE inhibition. After 12 weeks, the hemodynamic benefit was maintained. Omapatrilat may be a promising new agent in CHF. PMID- 12084606 TI - Prevalence and age-dependence of malignant mutations in the beta-myosin heavy chain and troponin T genes in hypertrophic cardiomyopathy: a comprehensive outpatient perspective. AB - OBJECTIVES: The goal of this study was to determine the prevalence of "malignant" mutations in hypertrophic cardiomyopathy (HCM). BACKGROUND: Previous genotype phenotype studies have implicated four mutations (R403Q, R453C, G716R and R719W) as highly malignant defects in the beta-myosin heavy chain (MYH7). In the cardiac troponin T gene (TNNT2), a specific mutation (R92W) has been associated with high risk of sudden death. Routine clinical screening for these malignant mutations has been suggested to identify high-risk individuals. METHODS: We screened 293 unrelated individuals with HCM seen at the Mayo Clinic in Rochester, Minnesota, between April 1997 and October 2000. Deoxyribonucleic acid (DNA) was obtained after informed consent; amplification of MYH7 exons 13 (R403Q), 14 (R453C) and 19 (G716R and R719W), and TNNT2 exon 9 (R92W) was performed by polymerase chain reaction. The mutations were detected using denaturing high-performance liquid chromatography and automated DNA sequencing. RESULTS: The mean age at diagnosis was 42 years with 53 patients diagnosed before age 25. The mean maximal left ventricular wall thickness was 21 mm. Nearly one-third of cases were familial and one-fourth had a family history of sudden cardiac death. Only 3 of the 293 patients possessed one of the five "malignant" mutations, and all 3 patients were <25 years of age at presentation (p < 0.006). CONCLUSIONS: This finding underscores the profound genetic heterogeneity in HCM. Only 1% of unrelated individuals seen at a tertiary referral center for HCM possessed one of the five "malignant" mutations that were examined. Routine clinical testing for these specific mutations is of low yield. PMID- 12084607 TI - Role of genotyping in risk factor assessment for sudden death in hypertrophic cardiomyopathy. PMID- 12084608 TI - Quantitation of basal dyssynchrony and acute resynchronization from left or biventricular pacing by novel echo-contrast variability imaging. AB - OBJECTIVES: This study sought to test a novel echocardiographic method based on contrast variability imaging (CVI), to quantify cardiac dyssynchrony and magnitude of resynchronization achieved by left ventricular (LV) and biventricular (BiV) pacing therapy. BACKGROUND: Left ventricular or BiV pacing is a promising new therapy for patients with heart failure and intraventricular conduction delay. However, precise quantitation of the extent of resynchronization achieved remains scant. METHODS: Ten patients treated with BiV or LV pacing therapy were studied. Echo-contrast was infused slowly, and gated images were acquired before and during contrast appearance. The temporally normalized variance derived from 30 to 50 sequential beats was determined at each pixel to yield the CVI image-displaying improved wall delineation. Systolic regional fractional area of radial sectors was calculated with active and temporarily suspended (AAI) pacing. All analyses were performed blinded to both patient and treatment. RESULTS: Pacing increased septal inward motion from -20.4 +/- 9.6% to -30.5 +/- 14.0%, whereas lateral wall motion occurred earlier with no net magnitude change. Both spatial and temporal dyssynchrony in the LV declined nearly 40% with LV or BiV pacing (p < or = 0.001), and this correlated with increasing ejection fraction (31% to 39%; p < 0.02; p < 0.004 for correlation with dyssynchrony). CONCLUSIONS: The new imaging and regional dyssynchrony analysis methods provide quantitative assessment of resynchronization analogous to that previously obtained only by tagged magnetic resonance imaging. This could provide a useful noninvasive method for both identifying candidates and following long-term therapy. PMID- 12084609 TI - Electrocardiogram-gated single-photon emission computed tomography versus cardiac magnetic resonance imaging for the assessment of left ventricular volumes and ejection fraction: a meta-analysis. AB - OBJECTIVES: The purpose of this study was to evaluate the accuracy of electrocardiogram (ECG)-gated single-photon emission computed tomography (SPECT) for assessment of left ventricular (LV) end-diastolic volume (EDV), end-systolic volume (ESV) and ejection fraction (EF) compared with the gold standard of cardiac magnetic resonance imaging (MRI). BACKGROUND: Several comparisons of ECG gated SPECT with cardiac MRI have been performed for evaluation of LV volumes and EF, but each has considered few subjects, thus leaving uncertainty about the frequency of discrepancies between the two methods. METHODS: We performed a meta analysis of data on 164 subjects from nine studies comparing ECG-gated SPECT versus cardiac MRI. Data were pooled in correlation and regression analyses relating ECG-gated SPECT and cardiac MRI measurements. The frequency of discrepancies of at least 30 ml in EDV, 20 ml in ESV and 5% or 10% in EF and concordance for EF < or =40% versus >40% were determined. RESULTS: There was an overall excellent correlation between ECG-gated SPECT and cardiac MRI for EDV (r = 0.89), ESV (r = 0.92) and EF (r = 0.87). However, rates of discrepancies for individual subjects were considerable (37% [95% confidence interval [CI], 26% to 50%] for at least 30 ml in EDV; 35% [95% CI, 23% to 49%] for at least 20 ml in ESV; 52% [95% CI, 37% to 63%] for at least 5% in EF; and 23% [95% CI, 11% to 42%] for at least 10% in EF). The misclassification rate for the 40% EF cutoff was 11%. CONCLUSIONS: Electrocardiogram-gated SPECT measurements of EDV, ESV and EF show high correlation with cardiac MRI measurements, but substantial errors may occur in individual patients. Electrocardiogram-gated SPECT offers useful functional information, but cardiac MRI should be used when accurate measurement is required. PMID- 12084611 TI - Cardiology workforce: there's already a shortage, and it's getting worse! PMID- 12084610 TI - Reverse remodeling and enhanced adrenergic reserve from passive external support in experimental dilated heart failure. AB - OBJECTIVES: We sought to test the efficacy of a passive elastic containment device to reverse chronic chamber remodeling and adrenergic down-regulation in the failing heart, yet still maintaining preload reserve. BACKGROUND: Progressive cardiac remodeling due to heart failure is thought to exacerbate underlying myocardial dysfunction. In a pressure-volume analysis, we tested the impact of limiting progressive cardiac dilation by an externally applied passive containment device on both basal and adrenergic-stimulated function in failing canine hearts. METHODS: Ischemic dilated cardiomyopathy was induced by repeated intracoronary microembolizations in six dogs. The animals were studied before and three to six months after surgical implantation of a thin polyester mesh (cardiac support device [CSD]) that surrounded both cardiac ventricles. Pressure-volume relations were measured by a conductance micromanometer catheter. RESULTS: Long term use of the CSD lowered end-diastolic and end-systolic volumes by -19 +/- 4% and -22 +/- 8%, respectively (both p < 0.0001) and shifted the end-systolic pressure-volume relation to the left (p < 0.01), compatible with reverse remodeling. End-diastolic pressure and chamber diastolic stiffness did not significantly change. The systolic response to dobutamine markedly improved after CSD implantation (55 +/- 8% rise in ejection fraction after CSD vs. -10 +/- 8% before CSD, p < 0.05), in conjunction with a heightened adenylyl cyclase response to isoproterenol. There was no change in the density or affinity of beta adrenergic receptors. Diastolic compliance was not adversely affected, and preload-recruitable function was preserved with the CSD, consistent with a lack of constriction. CONCLUSIONS: Reverse remodeling with reduced systolic wall stress and improved adrenergic signaling can be achieved by passive external support that does not generate diastolic constriction. This approach may prove useful in the treatment of chronic heart failure. PMID- 12084612 TI - Utility of a BNP as a marker for RV dysfunction in acute pulmonary embolism. PMID- 12084613 TI - The optimal "dose" of disease management programs in HF. PMID- 12084614 TI - Acetaminophen-induced suppression of hepatic AdoMet synthetase activity is attenuated by prodrugs of L-cysteine. AB - Administration of acetaminophen (ACP, 400 mg/kg, i.p.) to fasted, male Swiss Webster mice caused a rapid 90% decrease in total hepatic glutathione (GSH) and a 58% decrease in mitochondrial GSH by 2 h post ACP. This was followed by a time dependent decrease (72%) in hepatic AdoMet synthetase activity and rise in plasma ALT levels (>10000 U/l) at 24 h post ACP treatment. AdoMet synthetase activity was maintained at 82, 78 and 60% of controls, respectively, by the cysteine prodrugs PTCA, CySSME and NAC. Total hepatic and mitochondrial GSH levels were also protected from severe ACP-induced depletion by CySSME and MTCA. These results suggest that the maintenance of GSH homeostasis by cysteine prodrugs can protect mouse hepatic AdoMet synthetase, a sulfhydryl enzyme whose integrity is dependent on GSH, as well as the liver itself from the consequences of oxidative stress elicited by toxic metabolites of xenobiotics. PMID- 12084615 TI - Meso 2,3-dimercaptosuccinic acid (DMSA) and monoisoamyl DMSA effect on gallium arsenide induced pathological liver injury in rats. AB - The effect of meso 2,3-dimercaptosuccinic acid (DMSA) and monoisoamyl DMSA (MiADMSA) on gallium arsenide (GaAs) induced liver damage was studied. The oral feeding rat model was used in this study. The animals were exposed to 10 mg/kg GaAs, orally, once daily, 5 days a week for 24 weeks and treated thereafter with single oral daily dose of either 0.3 mmol/kg DMSA or MiADMSA for two course of 5 days treatment. The animals were sacrificed thereafter. Lipid peroxidation was assessed by measuring liver thiobarbituric acid reactive substance (TBARS). Liver damage was assessed by number of biochemical variables and by light microscopy. The activity of superoxide dismutase (SOD) and delta-aminolevulinic acid dehydratase (ALAD) beside reduced glutathione (GSH) concentration was measured in blood. Exposure to GaAs produced a significant reduction in GSH while, increased the oxidized glutathione (GSSG) concentration. Hepatic glutathione peroxidase (GPx) and catalase activity increased significantly while level of serum transaminase increased moderately. Gallium arsenide exposure also produced marked hepatic histopathological lesions. Overall, treatment with MiADMSA proved to be better than DMSA in the mobilization of arsenic and in the turnover of some of the above mentioned GaAs sensitive biochemical alterations. Histopathological lesions also, responded more favorably to chelation treatment with MiADMSA than DMSA. PMID- 12084616 TI - Syzygium cumini (Jamun) reduces the radiation-induced DNA damage in the cultured human peripheral blood lymphocytes: a preliminary study. AB - The effects of various concentrations (0.0, 1.56, 3.125, 6.25, 12.5, 25, 50 and 100 microg/ml) of the leaf extract of Syzygium cumini Linn. or Eugenia cumini (SC; black plum, Jamun, family Myrtaceae) was studied on the alteration in the radiation-induced micronuclei formation in the cultured human peripheral blood lymphocytes. Treatment of lymphocytes to various concentrations of SC resulted in a dose dependent increase in the micronuclei-induction, especially after 25-100 microg/ml extract. The exposure of human lymphocytes to various concentrations of SC extract before 3 Gy gamma-irradiation resulted in a significant decline in the micronuclei-induction at all the drug doses when compared with the non-drug treated irradiated cultures. A nadir in MNBNC frequency was observed for 12.5 microg/ml drug concentration, where the MNBNC frequency was approximately fourfold lower than that of the non-drug treated irradiated cultures. Therefore, this dose may be considered as an optimum dose for radiation protection. Our study demonstrates that the leaf extract of S. cumini, a plant traditionally used to treat diabetic disorders protects against the radiation-induced DNA damage. PMID- 12084617 TI - Acrolein causes transcriptional induction of phase II genes by activation of Nrf2 in human lung type II epithelial (A549) cells. AB - Acrolein, an alpha,beta-unsaturated aldehyde, is by far the strongest electrophile present in cigarette smoke which is involved in several lung pathophysiological conditions. Acrolein depletes glutathione and creates thiol imbalance. Acrolein due to thiol imbalance as well as covalent modification of cysteine is known to inhibit the activity of redox sensitive transcription factors such as NF-kappaB and AP-1. Exposure of human type II lung epithelial (A549) cells to non-lethal dose of acrolein (150 fmol/cell for 1 h) depletes 80% of intracellular glutathione and increases the transcription of gamma glutamylcysteine synthetase (gamma-GCS) at 6-12 h post-treatment, which helps in replenishing the glutathione to normal level. Acrolein treatment activates transcription of phase II genes in general, as indicated by an increase in mRNA for NAD (P) H:quinone oxidoreductase (NQO1). Western blot analysis revealed the increased level of the transcription factor, Nrf2 in the nuclear extract from acrolein treated cells. Electrophoretic mobility shift assay shows increased binding of nuclear proteins to human antioxidant response element (ARE) consensus sequence after treatment with acrolein. The involvement of Nrf2 in ARE mediated transcriptional activation in response to acrolein exposure has been confirmed by human NQO1-ARE reporter assay. The ability of acrolein to transcriptionaly activate genes responsible for phase II enzymes may form the basis of resistance against cell death and can have implications in cigarette smoke related lung carcinogenesis. PMID- 12084618 TI - Cytotoxicity by nitrite inhalants is not related to peroxynitrite formation. AB - Nitrite inhalant abuse has been correlated with HIV and Kaposi's sarcoma. Mouse models of inhalant exposure show immunosuppression and loss of immune cells. In the present study, isobutyl nitrite caused a dose-dependent loss of viability of a macrophage cell line. In the absence of cells, isobutyl nitrite reacted with hydrogen peroxide to form peroxynitrite. However, assays of mitochondrial respiration and nitration that detect peroxynitrite indicated that very little was present in cell cultures following exposure to the inhalants. Isobutyl, isoamyl, and butyl nitrites inhibited mitochondrial respiration, but only at high concentrations. Similarly, the nitrating activity of isobutyl nitrite occurred only at high concentrations and was not affected by the presence of hydrogen peroxide. Western blots showed that the inhalant did not increase nitrotyrosine formation in RAW cells or in peritoneal exudate macrophages (PEM) from exposed mice. Thus, the toxicity induced by isobutyl nitrite was probably not due to peroxynitrite formation. PMID- 12084619 TI - Amiodarone induces apoptosis in L-132 human lung epithelial cell line. AB - To investigate whether amiodarone induces apoptosis in cells of the L-132 human lung epithelial cell line, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay, 4,6-diamidino-2-phenylindole staining, DNA fragmentation assay, reverse transcription-polymerase chain reaction, and casapse-3 enzyme assay were performed. Through morphological and biochemical analyses, it was demonstrated that L-132 cells treated with amiodarone exhibit several features of apoptosis. In addition, it was shown that amiodarone increases the mRNA levels of bax and caspase-3. Based on the results, amiodarone appears to activate specific intracellular death-related pathways, including possibly the bax-dependent caspase-3 activation pathway, and thus induce apoptosis in human lung epithelial cells. PMID- 12084620 TI - Carp erythrocyte lipids as a potential target for the toxic action of zinc ions. AB - The effects of zinc on the lipid components of carp erythrocytes (Cyprinus carpio L.) were studied on red blood cells exposed to ZnSO4 (0.01-1 mM) for 20 h. The temperatures of the incubation medium were 5 and 20 degrees C for carp collected in winter and summer, respectively. It was observed that increase in the concentration of metal ions in the medium led to significant changes in the content of the main phospholipid classes. Zinc induced the reduction of phosphatidylcholine as well as the increase in the content of phosphatidylserine and phosphatidylinositol. A marked increase in the levels of malondialdehyde and conjugated dienes was also found both in the cells and the incubation medium. Moreover, elevated Zn concentrations caused alterations in the erythrocyte plasma membrane fluidity, estimated by measuring the fluorescence of 1-[4 trimethylaminophenyl]-6-phenyl-1,3,5-hexatriene and 12-[9-anthroyloxy]stearic acid, located on the surface and in the hydrophobic regions of the lipid bilayer. This study in vitro confirmed that higher concentrations of zinc ions induce marked changes in the composition and structure of membrane lipids in carp erythrocytes. PMID- 12084622 TI - Reduction of a 7,12-dimethylbenz[a]anthracene DNA adduct in rat mammary tissue in vivo when pretreated with tamoxifen. AB - Tamoxifen (TAM), an antiestrogenic compound, has been approved for the treatment of breast cancer in high risk women. TAM has been shown to be an effective agent for prevention of breast cancer in women of varying degrees of risk and has been proposed to be used prophylactically in women whose genetic background suggests a high risk for breast cancer. However, it is not known whether TAM given prophylactically will alter the response of women to carcinogens from common environmental exposures such as tobacco smoke. Therefore, we studied the effects of TAM pretreatment on mammary DNA adducts of the model carcinogen, 7,12 dimethylbenz[a]anthracene (DMBA), in the female rats to assess whether TAM would alter the adduct pattern of DMBA. TAM (0.3 mg/day) was given for 7 days prior to a single 20 mg DMBA gavage treatment that is considered a carcinogenic dose in the rat. At 7 days post-DMBA, there was a reduction in the major DMBA-DNA adduct and a significant reduction in the minor DMBA-DNA adduct in mammary glands (P=0.002) of TAM pretreated rats compared to control rats. These data indicate that TAM may alter either metabolic steps in the formation of DNA binding species and/or enhance adduct removal. These data suggest that TAM given to women prior to the development of breast cancer may modulate the impact of environmental exposures, for example, tobacco smoke. Furthermore, research is needed to determine if modulation will be positive, or negative as in the current study. PMID- 12084621 TI - Imidacloprid insecticide metabolism: human cytochrome P450 isozymes differ in selectivity for imidazolidine oxidation versus nitroimine reduction. AB - Many metabolites of imidacloprid (IMI) have been identified, but the enzymatic basis for their formation has not been reported. This study with individual recombinant cytochrome P450 (CYP450) isozymes from human liver shows that the principal organoextractable NADPH-dependent metabolites are the 5-hydroxy (major) and olefin (minor) derivatives from hydroxylation and desaturation of the imidazolidine moiety and the nitrosoimine (major), guanidine (minor) and urea (trace) derivatives from reduction and cleavage of the nitroimine substituent. Isozymes selective for imidazolidine oxidation in order of decreasing overall activity are CYP3A4>CYP2C19 or CYP2A6>CYP2C9, while those selective for nitroimine reduction are CYP1A2, CYP2B6, CYP2D6 and CYP2E1. Three flavin monooxygenase isozymes (FMO1, FMO3, and FMO5) with NADPH are not active as assayed. These observations establish site specificity in IMI metabolism by CYP450 isozymes and that a single enzyme (CYP3A4) both oxidizes and reduces IMI at the imidazolidine and nitroimine moieties, respectively. PMID- 12084624 TI - A cellular cofactor in HIV-1 assembly: INI1 is also an "outtie". AB - Recent findings have shed light on the previously poorly understood process by which HIV is assembled and released from infected cells. The host cell cofactor integrase interactor 1 (INI1), which was previously shown to chaperone the covalent linkage of the viral genome to the chromosomal DNA of the cell, also appears to assist in virus assembly. Another host cofactor, TSG101, that functions in vacuolar protein sorting has been found to play a key role in virus budding. PMID- 12084623 TI - Human congenital long QT syndrome: more than previously thought? AB - Mutations in KCNQ1 and KCNE1, the alpha- and beta-subunits of the I(KS) K+ channel, produce the cardiac long QT (LQT) syndrome. These subunits are expressed in heart and inner ear, but also in epithelial tissues such as kidney or intestine where their functional roles have remained elusive. Recent work has shown that KCNE1-deficient mice display chronic hypokalemia and hyperaldosteronism. These results have significant implications for human congenital LQT syndromes because hypokalemia increases the risk of ventricular arrhythmia and cardiac sudden death. PMID- 12084625 TI - Immunogenicity of rDNA-derived pharmaceuticals. AB - Many biopharmaceuticals have been reported to induce the production of antibodies. Several factors influence immunogenicity, including the methods of production, formulation and storage as well as patient characteristics (e.g. type of disease and genetic background). Because immunogenicity is unpredictable, testing for antibodies should always be part of the evaluation of new biopharmaceuticals, although the biological consequences of these antibodies are rarely severe. PMID- 12084629 TI - The pressing need for combined genotype-phenotype analysis in clinical practice. PMID- 12084630 TI - Can "real-lab" supersede "virtual-ligand" promises? PMID- 12084631 TI - Is DOPA a neurotransmitter? AB - Historically, 3,4-dihydroxyphenylalanine (DOPA) has been considered to be an inert amino acid that alleviates the symptoms of Parkinson's disease by its conversion to dopamine via the enzyme aromatic L-amino acid decarboxylase. In contrast to this generally accepted idea, we propose that DOPA itself is a neurotransmitter and/or neuromodulator in addition to being a precursor of dopamine. Several criteria such as synthesis, metabolism, active transport, existence, physiological release, competitive antagonism and physiological or pharmacological responses must be satisfied before a compound is accepted as a neurotransmitter. Recent evidence suggests that DOPA fulfills these criteria in its involvement in baroreflex neurotransmission. PMID- 12084632 TI - A model for the functioning of family 3 GPCRs. AB - Family 3 G-protein-coupled receptors (GPCRs) comprise the metabotropic glutamate (mglu) receptors, GABA(B) receptors, Ca(2+)-sensing receptors and some taste and putative pheromone receptors. All are composed of two domains, an extracellular ligand-binding domain and a transmembrane heptahelical domain that activates G proteins. Here we propose a model for the function of family 3 GPCRs that takes into account their structure. This model fits with specific pharmacological features of some family 3 GPCRs, such as modulation by positive and negative allosteric regulators. The model also reveals differences between GABA(B) receptors and Group I mglu receptors: in the former there is "tight" functional coupling between the two domains of the receptor whereas the "loose" coupling in the latter gives these receptors specific features not shared by many other GPCRs. PMID- 12084633 TI - The ligand paradox between affinity and efficacy: can you be there and not make a difference? AB - Efficacy is described in terms of the ligand-induced bias of receptor micro conformations. A virtual simulation of binding of a collection of random ligands can reconcile an apparent paradox in receptor theory, namely the theoretical requirement that affinity and efficacy be related to each other and the experimental fact that medicinal chemistry often can separate structure-activity relationships for affinity and efficacy. The simulation indicates that homoscedacity in the relationship allows for a general correlation with isolated instances of non-compliance. This leads to the conclusion that if a ligand binds to the receptor it will change the receptor by its presence. Therefore, this suggests that all ligands with macro-affinity should be extensively studied for pharmacological activities other than simple G-protein activation with the potential promise of discovering new therapeutic applications for old drugs. PMID- 12084634 TI - Cation-pi interactions in ligand recognition and catalysis. AB - The cation-pi interaction is a potent, general noncovalent binding force that is observed in a wide range of biological contexts. Here, we present an overview of well documented cases in which a cation-pi interaction makes an important contribution to small-molecule recognition at a protein binding site. From these and other studies it is clear that, in addition to the hydrophobic effect, hydrogen bonding and ion pairing, the cation-pi interaction must be considered when evaluating drug-receptor interactions. PMID- 12084635 TI - Does insulin dysfunction play a role in Alzheimer's disease? AB - Age-related changes in hormone levels are determinants of a variety of human diseases. Insulin is known to affect numerous brain functions including cognition and memory, and several clinical studies have established links between Alzheimer's disease (AD), insulin resistance and diabetes mellitus. These are reinforced by biological studies that reveal the effects of insulin on the molecular and cellular mechanisms that underlie the pathology of AD. For example, insulin regulates phosphorylation of tau protein, which underlies neurofibrillary lesions in the brains of AD patients. Insulin also affects the metabolism of beta amyloid, the main constituent of AD amyloid pathology. Here, we discuss clinical and biological data that highlight potential targets for therapeutic intervention. PMID- 12084636 TI - Teaching of pharmacology in the 21st century: new challenges and opportunities. PMID- 12084639 TI - Old principles revisited--traction splinting for closed proximal phalangeal fractures. AB - Proximal phalangeal fractures are commonly caused by a blunt injury to the dorsum of the hand. This typically results in a closed transverse fracture with volar angulation, which is an inherently unstable fracture pattern. K-wiring and internal fixation have the disadvantage of damage to the soft tissue envelope, risk of infection and wire loosening. We describe 15 patients treated with an alternative method of skin traction and splintage in our regional trauma centre. Fractures were reduced under X-ray and reduction was maintained using a combination of a malleable metal splint and tape to provide skin traction. Fourteen patients had an excellent objective and subjective outcome. The advantage of our technique is preservation of the soft tissue envelope and it should be considered an alternative to other treatment options as it is simple, quick, inexpensive and non-invasive. PMID- 12084640 TI - Percutaneous bone grafting in the treatment of the delayed union and non-union of tibial fractures. AB - The classic method of open bone grafting in the treatment of un-united tibial fractures has been associated with some complications. A novel, minimally invasive, percutaneous technique of bone grafting is described. Forty-one consecutive patients with delayed union or non-union of a tibial fracture was treated with percutaneous bone grafting from 1993 to 1999. The mean age of the patients was 41 (15-86) years. Twenty-seven patients had grade I-III open fractures. The fractures had initially been treated in a cast (n=26), external fixator (n=14) or intramedullary nail (n=1). Two patients had an earlier unsuccessful Phemister-Charnley-type bone-grafting procedure for delayed union. The mean interval from injury to percutaneous bone grafting was 21 (13-66) weeks. Thirty-seven of the 41 un-united fractures healed in 13 (10-48) weeks after the percutaneous bone grafting. The mean hospital stay was 1 day. No infections, bleeding or neural complications existed. Percutaneous bone grafting appeared to be as effective as open techniques, and possessed considerable advantages. It is safe, time saving and economical, it involves minimal trauma at the fracture site and it avoids major donor site problems. PMID- 12084641 TI - Intramedullary femoral nailing: removing the nail improves subjective outcome. AB - Intramedullary nailing has become an established treatment for femoral fractures. We reviewed the subjective and objective outcome of locked intramedullary nailing since its introduction to our unit in 1988. The outcome was assessed subjectively using the Short Form 36 (SF 36) health questionnaire and objectively by clinical, radiological and case note review. From 1988 to 1995, 220 intramedullary femoral nails were inserted at the North Staffordshire Royal Infirmary. Of these we were able to match the SF 36 questionnaire and case review of 91 patients. Fifty-six patients were male and 35 female with an average age of 40. Acute trauma accounted for 86 cases (six with an injury severity score >15 and seven open injuries). There was a 24% incidence of minor complications; there were no deep infections and no major complications. All cases recorded subjective scores within the normal range but 34 symptomatic patients who had their femoral nails removed (mainly for persistent pain or prominent metalwork) recorded noticeably higher scores in all eight assessment areas. Intramedullary nailing for femoral fractures is a technically demanding procedure and results in both a good objective and subjective outcome. It is associated with a low rate of major complications but a relatively high rate of minor complications. We conclude that the removal of intramedullary femoral nails is justified in symptomatic patients. It results in an improved subjective outcome and has a low complication rate. PMID- 12084642 TI - Traumatic injuries of the distal femoral physis. Retrospective study on 151 cases. AB - This is a retrospective study on a series of 151 injuries involving the distal femoral physis. The average age at the time of injury was 12.3 years. Patients were followed for an average of 8.2 years. The complications encountered were not insignificant and the satisfactory results were relatively low (64.9%). The juvenile age group was the most affected with complications. Salter-Harris type II injuries predominated (43.0%) and they did not have a good prognosis as previously suggested. Symptomatic knee ligamentous laxity was found in 12 patients (7.9%). Compartment syndrome is a devastating complication that occurred in 2 cases (1.3%). We stress the point that a child with a physeal injury of the distal femur should be followed for several years after injury and preferably until skeletal maturity. The surgeon must have high index of suspicious for compartment syndrome. Physeal injuries of the distal femur and in particular Salter and Harris type II should be reduced anatomically and fixed well. Whether this should be achieved by closed or opened means, requires a controlled prospective study to provide a confident answer. PMID- 12084643 TI - Outcomes following quadriceps tendon ruptures. AB - Complete rupture of the quadriceps femoris tendon is a well-described injury. There is a scarcity of literature relating to the outcome of patients with this injury after surgery. We undertook a retrospective analysis of patients who had surgical repair of their ruptured quadriceps tendon at our institution over a seven year period-totalling 27 patients. Males were more commonly affected with a M:F ratio of 8:1. The peak incidence was in the sixth decade of life. The non dominant limb was twice as likely to be affected. Assessment consisted of completion of a functional knee questionnaire, clinical examination and isokinetic testing on a Biodex dynanometer. Symptomatic outcome following surgical repair was excellent with a mean symptom score generated of 22.7 out of a maximum of 25. 18/19 patients returned to their premorbid level of activity on average 18.1 weeks following injury. There was no difference in quadriceps girth comparing affected and unaffected limbs. Less than 5 degrees deficit in range of motion existed between affected and unaffected limbs. Approximately two-thirds of patients were the same or better when comparing peak torque/body weight, average power, maximum average peak torque and total work/body weight in affected and unaffected limbs. PMID- 12084644 TI - A new method of removing a fractured interlocked nail. PMID- 12084645 TI - Augmentation of patellar tendon repair with poly-p-dioxannone cord. PMID- 12084646 TI - Distal radius fracture in a premature infant with osteopenia caused by handling during intravenous cannulation. PMID- 12084647 TI - Osteochondritis of the base of the first metacarpal following a physeal injury. PMID- 12084648 TI - A new technique for treatment of medial epicondyle fracture with displacement into the elbow joint. PMID- 12084649 TI - Isolated dislocation of the radial head, with simultaneous dislocation of proximal and distal radio-ulnar joints without fracture in an adult patient: case report and review of the literature. PMID- 12084650 TI - A case of forearm compartment syndrome following a ring avulsion injury. PMID- 12084651 TI - A case of intramyocardial sewing needle extracted without stopping the heart. PMID- 12084652 TI - Unilateral pringle manoeuvre and packing for haemostasis in severe liver trauma. PMID- 12084653 TI - Inferior thyroid artery injury due to airbag deployment. PMID- 12084654 TI - Hypotension as a delayed complication of rupture of a branch of the superior gluteal artery, following buttock contusion. PMID- 12084655 TI - Pathological fractures in primary hyperparathyroidism. A case report highlighting diagnostic difficulties. PMID- 12084656 TI - Fracture-dislocation of the ankle with fixed displacement of the fibula behind the tibia--a rare variant. PMID- 12084657 TI - Individual differences in children's cortisol response to the beginning of a new school year. AB - The relationship between salivary cortisol levels at the beginning of the school year and parent reports of temperament using the Children's Behavior Questionnaire (CBQ) were investigated in 35 first graders. Cortisol was sampled in the morning, afternoon, and evening on the first and fifth days of school and two weekend days later in the school year. On the first day of school compared to weekend days, children displayed a steeper cortisol slope across the day due to lower afternoon and evening levels. By the fifth day of school, cortisol levels, on average, were not different than on weekend days. However, using change scores to reflect the response to school days relative to weekend days, children who scored higher in Surgency (i.e., enjoyment of intense play, high activity level, impulsivity, lack of shyness) continued to display a steeper cortisol slope on the fifth day of school with higher morning and lower evening cortisol levels. In contrast, shyness, which negatively contributes to Surgency, was positively correlated with evening change scores on the fifth day of school. PMID- 12084658 TI - Differing concentrations of corticotropin-releasing factor and oxytocin in the cerebrospinal fluid of bonnet and pigtail macaques. AB - The two neuropeptides corticotropin-releasing-factor (CRF) and oxytocin (OT) may produce opposing behavioral effects - elevations of the former have been associated with anxiety and social vigilance and reductions of the latter with reduced social affiliation. We sought to test the hypothesis that, within the primate macaque genus, the more gregarious, affiliative, and affectively stable bonnet species (Macaca radiata) would exhibit lower cerebrospinal fluid (CSF) CRF and higher CSF OT concentrations in comparison to its close relative, the temperamentally volatile and socially distant pigtail (Macaca nemestrina). Cisternal CSF samples were obtained from young adult male and female pigtail and bonnet macaques, and CRF and OT concentrations were measured by radioimmunoassay. Pigtail macaques exhibited significantly higher concentrations of CSF CRF and significant lower concentrations of CSF OT than bonnet macaques. Results were not attributable to age or sex differences in group composition. When included together in a multiple regression, CRF and OT showed a multiple R of 0.76, accounting for more than half of the species variance. Although species differences in the bioeffectiveness of these peptides may possibly confound the observed biobehavioral relationships, in the absence of any existing data to that effect, the current findings appear in accordance with the hypothesis and consistent with previously reported species-typical behaviors observed in these macaques. PMID- 12084659 TI - REM sleep and prolactin in patients with non-affective psychoses. AB - Reduced REM latency is a common polysomnographic finding in patients with schizophrenia. This has been attributed to cholinergic hyperactivity secondary to increased dopaminergic tone. We studied polysomnographic sleep recordings, and morning serum prolactin levels as a measure of dopaminergic tone in 17 drug-free patients suffering from non-affective psychoses, hypothesizing that REM-latency and prolactin would correlate. A clear-cut positive correlation between prolactin and REM latency was found, as well as a negative correlation between prolactin and REM sleep. The findings may be explained by dopaminergic and secondary hypercholinergic and/or serotonergic mechanisms responsible for the regulation of REM sleep and the secretion of prolactin. PMID- 12084660 TI - Expression of tumor necrosis factor-alpha and interferon-gamma mRNA in blood cells correlates with depression scores during an acute attack in patients with multiple sclerosis. AB - Depression is a common problem in multiple sclerosis (MS) and affects about 50% of MS patients. Since a dysregulation of cytokine levels has been implicated in the pathogenesis of MS and alterations in cytokine serum levels have been found in depressive illness, we examined the relationship between depressive symptoms, cytokine mRNA expression levels of Th1-type and Th2-type cytokines and neurological disability among early diagnosed MS patients in a prospective study. Sixteen patients with clinically or laboratory supported MS were assessed using the Beck Depression Inventory (BDI) and the Kurtzke Expanded Disability Status Scale (EDSS). Cytokine mRNA in whole blood was serially determined by a new quantitative polymerase chain reaction (PCR) method. BDI sum scores (2,9 fold) and the expression levels of tumor necrosis factor-alpha (TNF-alpha; 4 fold), interferon-gamma (IFN-gamma; 4,6 fold) and interleukin-10 (IL-10; 6,1 fold) mRNA were increased in MS patients during an acute attack compared to age and sex matched healthy controls. We detected a significant positive correlation between TNF-alpha (r=0.55) and interferon-gamma (r=0.54) mRNA expression and the BDI sum scores during an acute attack in MS patients. At follow-up after 3-6 months, only TNF-alpha mRNA expression was correlated with BDI sum scores (r=0.62 resp. r=0.31). No correlation of the BDI sum scores with Th2-type cytokine mRNA expression for interleukin-4 (IL-4) and interleukin-10 (IL-10) or with the extent of neurological disability was observed. The possible contribution of Th1-type cytokines to the development of depression in MS is discussed. PMID- 12084661 TI - Pubertal changes in gonadal hormones do not underlie adolescent dopamine receptor overproduction. AB - Males, but not females, overproduce dopamine receptors in the striatum of rats across the periadolescent period followed by their elimination during young adulthood. In order to investigate the role that gonadal hormones play in this pubertal process, rats were castrated or ovariectomized at postnatal day (P) 28 when estrogen and testosterone levels are beginning to surge. Dopamine D1 and D2 striatal receptor density was then determined with autoradiography at P40 (adolescence) and P80 (young adulthood) to determine if either testosterone stimulates the overproduction of receptors in males or if estrogen inhibits this process in females. Neither castration nor ovariectomy altered dopamine receptor density, although enhanced testosterone levels increased D1 receptor binding 4.2% and 19.5% in males and females, respectively. The results of this study suggest that the endogenous rise in gonadal steroid hormones during puberty is not responsible for the overproduction of receptors in males or the lack of overproduction in females. PMID- 12084662 TI - HPA axis response to a psychological stressor in generalised social phobia. AB - Social phobia may be associated with a dysfunction of the hypothalamic-pituitary adrenal (HPA) axis. In this study we determined HPA axis responsivity to a psychological stressor in patients with social phobia and compared them to healthy controls. Fifteen patients with DSM IV social phobia with a mean score of 77.7 on the Liebowitz Social Anxiety Scale and 15 age and sex matched controls underwent the stressor consisting of mental arithmetic and a short term memory test performed in front of an audience. Plasma levels of cortisol and corticotropin were measured at various intervals throughout the test. Although baseline measures of cortisol did not differ between patients (319.8+/-34.6 nmol/l) and controls (279.5+/-42.7 nmol/l)(t=0.7, df=28, P<0.5) nor did baseline corticotropin values (8.6+/-2.1 pg/ml vs 13.7+/-2.0 pg/ml respectively) (t=-1.8, df=28, P<0.08) this stressor resulted in a significantly greater delta max cortisol response (the difference between baseline values and the maximum increase during the stressor) in patients (167.1+/-23.7 nmol/l) than in controls (106.7+/-16 nmol/l) (t=2.1, df=28, P<0.04). There was no significant difference in delta max corticotropin between groups (patients 8.8+/-2.1 pg/ml vs controls 9.1+/-1.9 pg/ml) (t=-0.08, df=28, P<0.9). This preliminary study indicates that patients with social phobia appear to have a hyper-responsive adrenocortical response to psychological stress. PMID- 12084663 TI - Corticotropin-releasing factor induces social preferences in male prairie voles. AB - Exposure to stressors facilitates the formation of social preferences in monogamous male prairie voles (Microtus ochrogaster). In the present study, the hypothesis was tested that treatment with corticotropin-releasing factor (CRF), a neuropeptide released during stress, is capable of inducing social preferences in male prairie voles. The effects of five doses of CRF (0.01, 0.1, 1.0, 10 and 100 ng; i.c.v.) on social preference were assessed. Exogenous CRF did not alter the amount of social contact that occurred between the experimental animal and partner during the initial cohabitation period. However, when tested after 3 h of cohabitation, animals that had been treated with 0.1 or 1.0 ng CRF spent significantly more time in physical contact with the partner than a stranger. In contrast, 3 h of cohabitation was not sufficient to induce social preferences in animals pre-treated with an artificial CSF vehicle or other doses of CRF. Furthermore, co-administration of a CRF receptor antagonist prevented the formation of CRF-induced social preferences. These data provide support for a role of the hypothalamic-pituitary-adrenal axis in social bonding in prairie voles. PMID- 12084665 TI - Attunement of maternal and child adrenocortical response to child challenge. AB - Although a great deal is known about physiological responding to stress in nonhuman animals, and also about individual differences in behavioral attunement in humans, physiological attunement between human mothers and their children has never been studied. The current study examined attunement in adrenocortical response between mother and child in the context of the child's exposure to a novel and potentially challenging task. Children ranging in age from two to four years of age walked on a balance beam for the first time while mothers watched on a monitor from the next room. Saliva samples were collected from both mothers and children before and 30 minutes following the beam walk. Individual differences in behavioral attunement were assessed from a videotaped mother-child teaching task, and coded for maternal sensitivity. We expected that mothers rated as highly sensitive would show better physiological attunement with their children's adrenocortical response to the balance beam walk than less sensitive mothers. We did not expect that all children would show a cortisol elevation in response to the task. Rather, we were interested in the degree to which mothers "matched" children's adrenocortical fluctuations, regardless of elevations or decreases in cortisol. Results supported the hypothesis. In the highly sensitive group, mothers' and children's adrenocortical responses to the child beam walk were significantly correlated, and in the less sensitive group the responses were not significantly related. Findings suggest that physiological attunement may co occur with behavioral sensitivity in normal mother-child relationships. PMID- 12084664 TI - Inhalation of 35% CO(2) results in activation of the HPA axis in healthy volunteers. AB - BACKGROUND: The hypothalamo-pituitary-adrenal (HPA) axis is a major stress responsive system in humans. Although there are numerous ways of testing responsiveness of the HPA in experimental animals, this is much more difficult in man. Hypercapnea is a very stressful stimulus for humans and has been used as an anxiogenic probe in psychiatric patients. We have now investigated whether the simple challenge of a single 35% inhalation of CO(2) activates the neuroendocrine system as evidenced by changes in HPA activity, as well as cardiovascular and subjective responses, in healthy volunteers. METHODS: Fourteen healthy male volunteers were recruited. They underwent single vital capacity inhalation of room air and 35% CO(2), in a single blind fashion. Neuroendocrine, cardiovascular and subjective fear measures were taken at regular intervals. RESULTS: CO(2) inhalation produced significant activation of the HPA axis in all subjects, as measured with plasma cortisol. Heart rate was decreased and systolic blood pressure was significantly increased shortly after the inhalation of CO(2). The subjects reported short-lived symptoms of fear with the experimental gas. CONCLUSIONS: Single vital capacity inhalation of 35% CO(2) activated the HPA axis in healthy volunteers. It also had a significant cardiovascular and psychological (anxiogenic) effect, as expected from previous published studies. The test is potentially useful in studying the responsivity of the HPA axis in health and disease. PMID- 12084666 TI - Short-term suppression of ovarian function and immune measures in healthy women. AB - Ovarian function was suppressed to postmenopausal levels with a GnRH agonist for four months in healthy, premenopausal women to examine the effect of short-term alterations in sex steroid exposure on immune measures at rest and in response to standardized laboratory stressors. Twenty-two women were assessed at three times: (a) during the early follicular phase of the menstrual cycle; (b) after ovarian function had been suppressed by a GnRH agonist; and (c) either in the early follicular phase after the resumption of regular menstrual cycling (group labeled Cycle), or when women received estradiol transdermally in conjunction with a GnRH agonist (group labeled Patch). A third group of 11 women was assessed at similar time points, but in a different order of pharmacologic intervention to evaluate the effects of ovarian function suppression that were not confounded by habituation to the laboratory stressors (group labeled Hormone Control). Finally, 15 women served as control subjects to control for the effects of time, seasonality and blood collection procedures on immune measures (group labeled Immune Control). Immune measures included circulating cell counts, response to mitogens (PHA and PWM), and NK cytotoxicity. Results revealed no consistent changes in basal or stress-induced immune measures that varied with a period of short-term suppression of ovarian function. Basal and acute stress-induced cell counts showed moderate to high temporal stability over a six to ten month time interval. PMID- 12084667 TI - Diet and cardiovascular disease. PMID- 12084670 TI - Industry attacks on science. PMID- 12084672 TI - Training the public health education workforce. PMID- 12084673 TI - Misunderstanding the World Health Report 2000. PMID- 12084677 TI - A code of ethics for public health. PMID- 12084678 TI - Critical policy challenges in the third decade of the HIV/AIDS epidemic. AB - Numerous policy challenges continue to face the United States in the third decade of the HIV/AIDS pandemic, in both the health and foreign policy arenas. They include long-standing questions about care, treatment, prevention, and research, as well as new ones introduced by the changing nature of the epidemic itself and the need to balance demands for limited resources. These challenges concern the United States not only in its role as a world leader in combating a global epidemic, but in its decisions and focus at home, where the epidemic continues to take a toll. PMID- 12084679 TI - Alternatives to a national system of population-based state cancer registries. AB - To support cancer control efforts, the National Program of Cancer Registries was established to enhance or develop cancer registries in every state. But there is an alternative. State cancer control programs could be adequately planned and evaluated without a cancer registry, and federal support of cancer registration could be selectively provided to registries that provide data needed to monitor cancer incidence and survival for the nation or that serve as a resource for population-based etiologic and cancer control research. The funds saved could be redirected to support the continued collection of high-quality cancer incidence and survival data for the nation as such efforts become more costly and complex in the future, and to expand support of population-based cancer research efforts. PMID- 12084680 TI - Time, place, and consciousness: three dimensions of meaning for US institutional review boards. AB - In the past few years, US federal agencies governing research with human subjects and institutional review boards have taken a higher-profile path than ever before, both at home and internationally. This trend carries profound significance for US-based institutions and has implications also for the rest of the world. What does this critical moment of heightened federal scrutiny mean for the workings of US institutional review boards? We examined board activity across 3 dimensions: time, place, and consciousness. We conclude that although institutions in all areas of biomedical and social science research are adapting their practices, the field of public health is especially well positioned to adapt to, and succeed in, new efforts to ensure protection of human research subjects. PMID- 12084681 TI - Nontherapeutic research with children: Grimes v Kennedy Krieger Institute. AB - Research with young children raises difficult issues of law and ethics. A recent Maryland case, Grimes v Kennedy Krieger Institute, Inc, appears to impose restrictive rules on research with children when the subjects are put at risk but cannot derive direct benefit from their involvement in the research project. This case exemplifies the tension that exists between the goal of science to increase knowledge and the protection of the rights and welfare of nonconsenting research subjects. While some language in the opinion may be difficult to understand or apply, for the most part the case reflects the problems other courts and ethicists have had in delineating the role of children in "nontherapeutic" research. PMID- 12084682 TI - Risk and responsibility: ethics, Grimes v Kennedy Krieger, and public health research involving children. AB - The legal case of Grimes v. Kennedy Krieger Institute, Inc, has raised concerns in the public health research community regarding the acceptable level of risk in research involving children, parental authority for informed consent, and exploitation of research subjects for the benefit of public health. We provide an overview of the case and discuss the impact of the court's decision and its possible effect on future research protection policies and practices. PMID- 12084683 TI - The impact of genomics on global health. AB - Ensuring that advances in genomics are applied to the health improvement of people living in developing countries is an important contemporary challenge. In the near term, such advances are likely to alleviate infectious diseases, with longer-term benefits envisaged for chronic disorders. To ensure that benefits are shared by developing countries, attention must be paid to complex ethical, legal, social, and economic issues, as well as to public education and engagement. Creative and equitable international mechanisms and goodwill are needed to turn high hopes into reality and allow the use of genomics to reduce health inequities between rich and poor nations. PMID- 12084685 TI - The effects of the corset. PMID- 12084684 TI - Ethical oversight of public health research: can rules and IRBs make a difference in developing countries? AB - Controversies in the conduct of international research continue to pose challenges for the system of ethical review, particularly for developing countries. Although the concept of vulnerability is key to addressing these challenges, ethical review has typically ignored the agency of vulnerable participants and groups in determining what kind of review process is needed. Concurrent with developments shaping the new public health that seek to operationalize empowerment of communities by placing them as initiators and organizers of their own health, ethical review of public health research must find ways to recognize the agency of vulnerable individuals, groups, and communities in the review process if it is to address effectively the ethical dilemmas currently evident in collaborative international research. PMID- 12084686 TI - Burning Love: big tobacco takes aim at LGBT youths. AB - Secret tobacco industry documents lay bare the industry's targeting, seduction, and recruitment of minority groups and children. They also unmask Big Tobacco's disdain for its targets. PMID- 12084687 TI - Increasing the use of child restraints in motor vehicles in a Hispanic neighborhood. AB - The use of restraints in motor vehicles is less common in minority and low-income populations than in the general population. A preliminary survey of Hispanic preschool-aged children in west Dallas, Tex, conducted in 1997 showed much lower child restraint use (19% of those surveyed) than among preschool children of all races in the rest of the city (62%). Because there are few reports of successful programs to increase child restraint use among Hispanics, we undertook to implement and evaluate such a program. The program was conducted by bilingual staff and was tailored for this community. It was successful in increasing both child restraint use and driver seat belt use. PMID- 12084688 TI - Zip code caveat: bias due to spatiotemporal mismatches between zip codes and US census-defined geographic areas--the Public Health Disparities Geocoding Project. PMID- 12084689 TI - Rural-urban differences in the public health workforce: local health departments in 3 rural Western states. PMID- 12084690 TI - Determinants of maternal vitamin A or beta-carotene supplementation coverage: village-based female distributors in Nepal. PMID- 12084691 TI - Prevalence of exclusive breastfeeding among US infants: the Third National Health and Nutrition Examination Survey (Phase II, 1991-1994). PMID- 12084692 TI - Sexual and drug use behavior among women who have sex with both women and men: results of a population-based survey. PMID- 12084693 TI - A tailored intervention to aid decision-making about hormone replacement therapy. PMID- 12084694 TI - Effects of hospital staffing and organizational climate on needlestick injuries to nurses. AB - OBJECTIVES: This study determined the effects of nurse staffing and nursing organization on the likelihood of needlestick injuries in hospital nurses. METHODS: We analyzed retrospective data from 732 and prospective data from 960 nurses on needlestick exposures and near misses over different 1-month periods in 1990 and 1991. Staffing levels and survey data about working climate and risk factors for needlestick injuries were collected on 40 units in 20 hospitals. RESULTS: Nurses from units with low staffing and poor organizational climates were generally twice as likely as nurses on well-staffed and better-organized units to report risk factors, needlestick injuries, and near misses. CONCLUSIONS: Staffing and organizational climate influence hospital nurses' likelihood of sustaining needlestick injuries. Remedying problems with understaffing, inadequate administrative support, and poor morale could reduce needlestick injuries. PMID- 12084695 TI - Prescription noncompliance due to cost among adults with disabilities in the United States. AB - OBJECTIVES: This study estimated national prevalence rates of medication noncompliance due to cost and resulting health problems among adults with disabilities. METHODS: Analyses involved 25,805 respondents to the Disability Follow-Back Survey, a supplement to the 1994 and 1995 National Health Interview Surveys. RESULTS: Findings showed that about 1.3 million adults with disabilities did not take their medications as prescribed because of cost, and more than half reported health problems as a result. Severe disability, poor health, low income, lack of insurance, and a high number of prescriptions increased the odds of being noncompliant as a result of cost. CONCLUSIONS: Prescription noncompliance due to cost is a serious problem for many adults with chronic disease or disability. Most would not be helped by any of the current proposals to expand Medicare drug coverage. PMID- 12084696 TI - Twenty years of public health research: inclusion of lesbian, gay, bisexual, and transgender populations. AB - OBJECTIVES: This study determined to what extent lesbian, gay, bisexual, and transgender (LGBT) populations have been studied over the past 20 years of public health research. METHODS: From MEDLINE English-language articles on human subjects published between 1980 and 1999, I identified articles that included LGBT individuals. The abstracts were analyzed with a coding procedure that categorized the content by topic, sexual orientation, and race/ethnicity. RESULTS: LGBT issues were addressed by 3777 articles, or 0.1% of all Medline articles; 61% of the articles were disease-specific, and 85% omitted reference to race/ethnicity. Research unrelated to sexually transmitted diseases addressed lesbians and gay men with similar frequency, whereas bisexual persons were less frequently considered, and the least amount of research focused on transgender individuals. CONCLUSIONS: Findings supported that LGBT issues have been neglected by public health research and that research unrelated to sexually transmitted diseases is lacking. PMID- 12084697 TI - Prediction of depressive distress in a community sample of women: the role of sexual orientation. AB - OBJECTIVES: This study compared factors known or hypothesized to influence depressive symptomatology in a community sample of lesbians and heterosexual women. METHODS: Data were collected in a multisite survey of lesbians' physical and mental health. RESULTS: Findings confirmed earlier reports suggesting that traumatic life events such as physical and sexual abuse, and individual traits and coping styles are risk factors for depressive distress. However, findings of higher rates of suicidal behavior and of several risk factors for depressive distress among lesbians suggest that risk for depression may differ among lesbians and heterosexual women. CONCLUSIONS: Sexual orientation may represent an important but poorly understood risk factor for depressive distress as well as suicidal ideation and behavior. PMID- 12084698 TI - Risk behaviors, medical care, and chlamydial infection among young men in the United States. AB - OBJECTIVES: This study assessed factors related to chlamydial infection among young men in the United States. METHODS: Data were from interviews of nationally representative samples of 470 men aged 18 to 19 years (teenagers) and 995 men aged 22 to 26 years (young adults) and from urine specimens tested by means of polymerase chain reaction. RESULTS: Although a majority of the men reported occasional unprotected intercourse, only a minority perceived themselves to be at risk for contracting a sexually transmitted disease (STD). Chlamydial infection was detected in 3.1% of the teenagers and 4.5% of the young adults. A minority of those infected had symptoms or had been tested for STDs; very few had been diagnosed with STDs. CONCLUSIONS: Chlamydial infection is common but usually asymptomatic and undiagnosed. Primary and secondary prevention efforts should be increased, particularly among young adult men. PMID- 12084699 TI - Mammography screening and differences in stage of disease by race/ethnicity. AB - OBJECTIVES: We examined the effect of routine screening on breast cancer staging by race/ethnicity. METHODS: We used a 1990 to 1998 mammography database (N = 5182) of metropolitan Denver, Colo, women to examine each racial/ethnic cohort's incident cancer cases (n = 1902) and tumor stage distribution given similar patterns of routine screening use. RESULTS: Regardless of race/ethnicity, women participating in routine screenings had earlier-stage disease by 5 to 13 percentage points. After control for possible confounding factors, White women were more likely to have early-stage disease compared with Black and Hispanic women. CONCLUSIONS: Lack of screening coverage in certain racial/ethnic populations has often been cited as a reason for tumor stage differences at detection. In this study, correcting for screening did not completely reduce stage differentials among Black and Hispanic women. PMID- 12084700 TI - Optimal indicators of socioeconomic status for health research. AB - OBJECTIVES: In this study we examined the relationship between indicators of socioeconomic status (SES) and mortality for a representative sample of individuals. METHODS: The sample included 3734 individuals aged 45 and older interviewed in 1984 in the Panel Study of Income Dynamics. In the current study, mortality was tracked between 1984 and 1994 and is related to SES indicators of education, occupation, income, and wealth. RESULTS: Wealth and recent family income were the indicators that were most strongly associated with subsequent mortality. These associations persisted after we controlled for the other SES indicators and were stronger for women than for men and for non-elderly than for elderly individuals. CONCLUSIONS: We found that the economic indicators of SES were usually as strongly associated with mortality as, if not more strongly associated with mortality than, the more conventional indicators of completed schooling and occupation. PMID- 12084701 TI - Checking the pulse: Midwestern reporters' opinions on their ability to report health care news. AB - OBJECTIVES: Newspapers play a key role in disseminating information and shaping perceptions about health, research, and policies. Inadequate or misleading reporting constitutes a public health threat that can jeopardize individual health and lead to harmful health policies. METHODS: Surveys were mailed to 165 reporters at 122 newspapers in 5 Midwest states. The association of training, newspaper size, and experience with reporter's self-perceived reporting ability was assessed. RESULTS: The response rate was 69.6% (115/165). Between 66% and 85% of the reporters assessed 4 tasks vital to sound health reporting as "sometimes difficult" to "nearly always difficult." No significant differences in perceived ability were found by training or newspaper size. Respondents with less experience reported higher perceived ability. CONCLUSIONS: These findings show that reporters may have difficulty understanding complex health issues and interpreting statistics because they are inadequately trained. PMID- 12084702 TI - Increasing rural-urban gradients in US suicide mortality, 1970-1997. AB - OBJECTIVES: This study examined rural-urban gradients in US suicide mortality and the extent to which such gradients varied across time, sex, and age. METHODS: Using a 10-category rural-urban continuum measure and 1970-1997 county mortality data, we estimated rural-urban differentials in suicide mortality over time by multiple regression and Poisson regression models. RESULTS: Significant rural urban gradients in age-adjusted male suicide mortality were found in each time period, indicating rising suicide rates with increasing levels of rurality. The gradient increased consistently, suggesting widening rural-urban differentials in male suicides over time. When controlled for geographic variation in divorce rate and ethnic composition, rural men, in each age cohort, had about twice the suicide rate of their most urban counterparts. Observed rural-urban differentials for women diminished over time. In 1995 to 1997, the adjusted suicide rates for young and working-age women were 85% and 22% higher, respectively, in rural than in the most urban areas. CONCLUSIONS: The slope of the relationship between rural urban continuum and suicide mortality varied substantially by time, sex, and age. Widening rural-urban disparities in suicide may reflect differential changes over time in key social integration indicators. PMID- 12084703 TI - Assessment of a new approach to family planning services in rural Pakistan. AB - OBJECTIVES: In 1993, the government of Pakistan started a new approach to the delivery of contraceptive services by training literate married women to provide doorstep advice and supplies in their own and neighboring communities. This report assesses whether this community-based approach is starting to have an impact on contraceptive use in rural areas. METHODS: A clustered nationally representative survey was used to collect data on contraceptive use and access to services in each cluster. Two-level logistic regression was applied to assess the effects of service access, after potential confounders were taken into account. RESULTS: Married women living within 5 km of 2 community-based workers were significantly more likely to be using a modern, reversible method of contraception than those with no access (odds ratio = 1.74; 95% confidence interval = 1.11, 2.71). CONCLUSIONS: After decades of failure, the managers of the family planning program have designed a way of presenting modern contraceptives that is appropriate to the conditions of rural Pakistan. The new community-based approach should be steadily expanded. PMID- 12084704 TI - Use of topical sunscreens and the risk of malignant melanoma: a meta-analysis of 9067 patients from 11 case-control studies. AB - OBJECTIVES: This study examined the methodology of epidemiological studies that suggest use of topical sunscreen preparations is associated with increased risk of malignant melanoma. METHODS: We pooled data from observational studies using a general variance-based meta-analytic method that employed confidence intervals (previously described). The outcome of interest was a summary relative risk (RR) reflecting the risk of melanoma associated with sunscreen use versus nonuse. Sensitivity analyses were performed when necessary to explain any observed statistical heterogeneity. RESULTS: Combining studies that used non-heterogeneous data yielded a summary RR of 1.01, indicating no association between sunscreen use and development of malignant melanoma. CONCLUSIONS: The available epidemiological data do not support the existence of a relationship between topical sunscreen use and an increased risk of cutaneous malignant melanoma. PMID- 12084705 TI - The perceived impact of privatization on local health departments. AB - OBJECTIVES: This article presents nationally representative data on the effects of privatization on local health departments (LHDs). METHODS: A stratified representative national sample of 380 LHDs was drawn from a national list of 2488 departments. Telephone interviews were conducted with 347 LHD directors. RESULTS: One half of the directors of LHDs with privatized services reported that privatization helped the performance of core functions. Privatization often resulted in increased time needed for management and administration. More than a third of LHD directors reported concern about loss of control over the performance of privatized functions and services. CONCLUSIONS: Privatization is part of a broader shift toward "managing" rather than directly providing public health services, yet privatization often reduces LHDs' control over the performance of services. PMID- 12084706 TI - Wengen, a member of the Drosophila tumor necrosis factor receptor superfamily, is required for Eiger signaling. AB - We identified Wengen, the first member of the Drosophila tumor necrosis factor receptor (TNFR) superfamily. Wengen is a type III membrane protein with conserved cysteine-rich residues (TNFR homology domain) in the extracellular domain, a hallmark of the TNFR superfamily. wengen mRNA is expressed at all stages of Drosophila development. The small-eye phenotype caused by an eye-specific overexpression of a Drosophila TNF superfamily ligand, Eiger, was dramatically suppressed by down-regulation of Wengen using RNA interference. In addition, Wengen and Eiger physically interacted with each other through their TNFR homology domain and TNF homology domain, respectively. These results suggest that Wengen can act as a component of a functional receptor for Eiger. Our identification of Wengen and further genetic analysis should provide increased understanding of the evolutionarily conserved roles of TNF/TNFR superfamily proteins in normal development, as well as in some pathophysiological conditions. PMID- 12084707 TI - Regulatory motifs for CREB-binding protein and Nfe2l2 transcription factors in the upstream enhancer of the mitochondrial uncoupling protein 1 gene. AB - Thermogenesis against cold exposure in mammals occurs in brown adipose tissue (BAT) through mitochondrial uncoupling protein (UCP1). Expression of the Ucp1 gene is unique in brown adipocytes and is regulated tightly. The 5'-flanking region of the mouse Ucp1 gene contains cis-acting elements including PPRE, TRE, and four half-site cAMP-responsive elements (CRE) with BAT-specific enhancer elements. In the course of analyzing how these half-site CREs are involved in Ucp1 expression, we found that a DNA regulatory element for NF-E2 overlaps CRE2. Electrophoretic mobility shift assay and competition assays with the CRE2 element indicates that nuclear proteins from BAT, inguinal fat, and retroperitoneal fat tissue interact with the CRE2 motif (CGTCA) in a specific manner. A supershift assay using an antibody against the CRE-binding protein (CREB) shows specific affinity to the complex from CRE2 and nuclear extract of BAT. Additionally, Western blot analysis for phospho-CREB/ATF1 shows an increase in phosphorylation of CREB/ATF1 in HIB-1B cells after norepinephrine treatment. Transient transfection assay using luciferase reporter constructs also indicates that the two half-site CREs are involved in transcriptional regulation of Ucp1 in response to norepinephrine and cAMP. We also show that a second DNA regulatory element for NF-E2 is located upstream of the CRE2 region. This element, which is found in a similar location in the 5'-flanking region of the human and rodent Ucp1 genes, shows specific binding to rat and human NF-E2 by electrophoretic mobility shift assay with nuclear extracts from brown fat. Co-transfections with an Nfe2l2 expression vector and a luciferase reporter construct of the Ucp1 enhancer region provide additional evidence that Nfe2l2 is involved in the regulation of Ucp1 by cAMP-mediated signaling. PMID- 12084708 TI - Signal transduction through tyrosine-phosphorylated C-terminal fragments of amyloid precursor protein via an enhanced interaction with Shc/Grb2 adaptor proteins in reactive astrocytes of Alzheimer's disease brain. AB - The proteolytic processing of amyloid precursor protein (APP) through the formation of membrane-bound C-terminal fragments (CTFs) and of soluble beta amyloid peptides likely influences the development of Alzheimer's disease (AD). We show that in human brain a subset of CTFs are tyrosine-phosphorylated and form stable complexes with the adaptor protein ShcA. Grb2 is also part of these complexes, which are present in higher amounts in AD than in control brains. ShcA immunoreactivity is also greatly enhanced in patients with AD and occurs at reactive astrocytes surrounding cerebral vessels and amyloid plaques. A higher amount of phospho-ERK1,2, likely as result of the ShcA activation, is present in AD brains. In vitro experiments show that the ShcA-CTFs interaction is strictly confined to glial cells when treated with thrombin, which is a well known ShcA and ERK1,2 activator and a regulator of APP cleavage. In untreated cells ShcA does not interact with either APP or CTFs, although they are normally generated. Altogether these data suggest that CTFs are implicated in cell signaling via Shc transduction machinery, likely influencing MAPK activity and glial reaction in AD patients. PMID- 12084709 TI - Pctaire1 interacts with p35 and is a novel substrate for Cdk5/p35. AB - Cyclin-dependent kinase 5 (Cdk5) is a serine/threonine kinase that plays important roles during central nervous system development. Cdk5 kinase activity depends on its regulatory partners, p35 or p39, which are prominently expressed in the central nervous system. We have previously demonstrated the involvement of Cdk5 in the regulation of acetylcholine receptor expression at the neuromuscular junction, suggesting a novel functional role of Cdk5 at the synapse. Here we report the identification of Pctaire1, a member of the Cdk-related kinase family, as a p35-interacting protein in muscle. Binding of Pctaire1 to p35 can be demonstrated by in vitro binding assay and co-immunoprecipitation experiments. Pctaire1 is associated with p35 in cultured myotubes and skeletal muscle, and is concentrated at the neuromuscular junction. Furthermore, Pctaire1 can be phosphorylated by the Cdk5/p25 complex, and serine 95 is the major phosphorylation site. In brain and muscle of Cdk5 null mice, Pctaire1 activity is significantly reduced. Moreover, Pctaire1 activity is increased following preincubation with brain extracts and phosphorylation by the Cdk5/p25 complex. Taken together, our findings demonstrate that Pctaire1 interacts with p35, both in vitro and in vivo, and that phosphorylation of Pctaire1 by Cdk5 enhances its kinase activity. PMID- 12084710 TI - Phorbol ester-regulated oligomerization of diacylglycerol kinase delta linked to its phosphorylation and translocation. AB - Diacylglycerol kinase (DGK) plays an important role in signal transduction through modulating the balance between two signaling lipids, diacylglycerol and phosphatidic acid. In yeast two-hybrid screening, we unexpectedly found a self association of the C-terminal part of DGKdelta containing a sterile alpha-motif (SAM) domain. We then bacterially expressed the SAM domain fused with maltose binding protein and confirmed the formation of dimeric and tetrameric structures. Moreover, gel filtration and co-immunoprecipitation analyses demonstrated that DGKdelta formed homo-oligomeric structures in intact cells and that the SAM domain was critically involved in the oligomerization. Interestingly, phorbol ester stimulation induced dissociation of the oligomeric structures with concomitant phosphorylation of DGKdelta. Furthermore, we found that DGKdelta was translocated from cytoplasmic vesicles to the plasma membrane upon phorbol ester stimulation. In this case, DGKdelta mutants lacking the ability of self association were localized at the plasma membranes even in the absence of phorbol ester. A protein kinase C inhibitor, staurosporine, blocked all of the effects of phorbol ester, i.e. oligomer dissociation, phosphorylation, and translocation. We confirmed that tumor-promoting phorbol esters did not directly bind to DGKdelta. The present studies demonstrated that the formation and dissociation of oligomers serve as the regulatory mechanisms of DGKdelta and that DGKdelta is a novel downstream effector of phorbol ester/protein kinase C signaling pathway. PMID- 12084711 TI - The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induces apoptosis in mouse nigrostriatal glia. Relevance to nigral neuronal death and striatal neurochemical changes. AB - Swiss mice were given 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), 25 mg/kg/day, for 5 consecutive days and killed at different days after MPTP discontinuance. Decreases in striatal tyrosine hydroxylase activity and levels of dopamine and its metabolites were observed 1 day after MPTP discontinuance. Ascorbic acid and glutamate levels had increased, dehydroascorbic acid and GSH decreased, whereas catabolites of high-energy phosphates (inosine, hypoxanthine, xanthine, and uric acid) were unchanged. In addition, gliosis was observed in both striatum and substantia nigra compacta (SNc). Sections of SNc showed some terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end labeling (TUNEL)-positive cells. Neurochemical parameters of dopaminergic activity showed a trend toward recovery 3 days after MPTP discontinuance. At this time point, TUNEL-positive cells were detected in SNc; some of them showed nuclei with neuronal morphology. A late (days 6-11) increase in striatal dopamine oxidative metabolism, ascorbic acid oxidative status, and catabolites of high-energy phosphates were observed concomitant with nigral neuron and nigrostriatal glial cell apoptotic death, as revealed by TUNEL, acridine orange, and Hoechst staining, and transmission electron microscopy. These data suggest that MPTP-induced activation/apoptotic death of glial cells plays a key role in the sequential linkage of neurochemical and cellular events leading to dopaminergic nigral neuron apoptotic death. PMID- 12084712 TI - Mus81-Eme1 and Rqh1 involvement in processing stalled and collapsed replication forks. AB - The processing of stalled replication forks and the repair of collapsed replication forks are essential functions in all organisms. In fission yeast DNA junctions at stalled replication forks appear to be processed by either the Rqh1 DNA helicase or Mus81-Eme1 endonuclease. Accordingly, we show that the hypersensitivity to agents that cause replication fork stalling of mus81, eme1, and rqh1 mutants is suppressed by a Holliday junction resolvase (RusA), as is the synthetic lethality of a mus81(-) rqh1(-) double mutant. Recombinant Mus81-Eme1, purified from Escherichia coli, readily cleaves replication fork structures but cleaves synthetic Holliday junctions relatively poorly in vitro. From these data we propose that Mus81-Eme1 can process stalled replication forks before they have regressed to form a Holliday junction. We also implicate Mus81-Eme1 and Rqh1 in the repair of collapsed replication forks. Here Mus81-Eme1 and Rqh1 seem to function on different substrates because RusA can substitute for Mus81-Eme1 but not Rqh1. PMID- 12084713 TI - Pharmacology and functional properties of NTS2 neurotensin receptors in cerebellar granule cells. AB - The binding and signaling properties of neuronal NTS2 neurotensin (NT) receptors were examined in cultured rat cerebellar granule cells. As shown by reverse transcription-PCR, receptor autoradiography, and confocal microscopic localization of fluorescent NT, these cells selectively express the NTS2 receptor subtype. Accordingly, a single apparent class of (125)I-NT-binding sites, with an affinity of 3.1 nm, was detected in cerebellar granule cell cultures. This binding was competed for with high affinity (IC(50) = 5.7 nm) by the NTS2 ligand levocabastine and with low affinity (IC(50) = 203 nm) by the NTS1 antagonist SR48692. Hypertonic acid stripping of surface-bound ligand and hyperosmolar sucrose treatment revealed that 64% of specifically bound (125)I-NT was internalized at equilibrium via a clathrin-dependent pathway. In cells loaded with the Ca(2+)-sensitive fluorescent dye Fluo4, SR48692, but neither NT nor levocabastine, triggered a marked increase in cytosolic [Ca(2+)](i). By contrast, both NT and levocabastine, but not SR48692, induced a sustained (>60 min) activation of the mitogen-activated protein kinases, p42/p44, indicating functional coupling of NTS2 receptors. Complementary experiments carried out on synaptosomes from adult rat cerebellum demonstrated the presence of presynaptic NTS2 receptors. However, in contrast to perikaryal NTS2 sites, these presynaptic receptors did not internalize in response to NT stimulation. Taken together, the present results demonstrate that NTS2 receptors are present both presynaptically and postsynaptically in central neurons and that NT and levocabastine act as agonists on these receptors. PMID- 12084715 TI - DnaK-sigma 32 interaction is temperature-dependent. Implication for the mechanism of heat shock response. AB - The heat shock response in bacteria is a complex phenomenon in which sigma 32 plays the central role. The DnaK/J chaperone system binds and promotes degradation of sigma 32 at lower temperatures. At heat shock temperatures, the DnaK/J-mediated degradation of sigma 32 is largely abolished by a mechanism, which is not yet fully understood. In this article we have shown that interaction of DnaK with sigma 32 is highly temperature-dependent. This interaction is completely abolished at 42 degrees C. To investigate the origin of such strong temperature dependence, we have monitored the structural changes that occur in the sigma 32 protein upon upshift of temperature and attempted to elucidate its functional roles. Upon a shift of temperature from 30 to 42 degrees C, the CD spectrum of sigma 32 becomes significantly more positive without significant change in either tryptophan fluorescence spectra or quenchability to external quenchers. 1,8-Anilinonaphthalene sulfonic acid binding at 42 degrees C is not significantly affected. The equilibrium guanidine hydrochloride denaturation of sigma 32 is biphasic. The first phase shifts to even lower guanidine hydrochloride concentrations at 42 degrees C, whereas the major phase remains largely unchanged. The sigma 32-core interaction remains unchanged as a function of temperature. This suggests that increased temperature destabilizes a structural element. We discuss the possible location of this temperature sensitive structural element. PMID- 12084714 TI - Raloxifene, a mixed estrogen agonist/antagonist, induces apoptosis through cleavage of BAD in TSU-PR1 human cancer cells. AB - Selective estrogen receptor modulator is a proven agent for chemoprevention and chemotherapy of cancer. Raloxifene, a mixed estrogen agonist/antagonist, was developed to prevent osteoporosis and potentially reduce the risk of breast cancer. In this study, we examined the effect of raloxifene on the TSU-PR1 cell line. This cell line was originally reported to be a prostate cancer cell line, but recently it has been shown to be a human bladder transitional cell carcinoma cell line. The TSU-PR1 cell line contains high levels of estrogen receptor beta. Following treatment with raloxifene, evidence of apoptosis, including change in nuclear morphology, DNA fragmentation, and cytochrome c release, was observed in a dose-dependent manner in the TSU-PR1 cells (10(-9) to 10(-6) m range). We observed no detectable change in the steady-state levels of Bax, Bcl-2, and Bcl X(L) following raloxifene treatment. However, raloxifene induced caspase dependent cleavage of BAD to generate a 15-kDa truncated protein. Overexpression of a double mutant BAD resistant to caspase 3 cleavage blocked raloxifene-induced apoptosis. These results demonstrate that raloxifene induces apoptosis through the cleavage of BAD in TSU-PR1 cells. This molecular mechanism of apoptosis suggests that raloxifene may be a therapeutic agent for human bladder cancer. PMID- 12084716 TI - Copper-dependent autocleavage of glypican-1 heparan sulfate by nitric oxide derived from intrinsic nitrosothiols. AB - Cell surface heparan sulfate proteoglycans facilitate uptake of growth-promoting polyamines (Belting, M., Borsig, L., Fuster, M. M., Brown, J. R., Persson, L., Fransson, L.-A., and Esko, J. D. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 371 376). Increased polyamine uptake correlates with an increased number of positively charged N-unsubstituted glucosamine units in the otherwise polyanionic heparan sulfate chains of glypican-1. During intracellular recycling of glypican 1, there is an NO-dependent deaminative cleavage of heparan sulfate at these glucosamine units, which would eliminate the positive charges (Ding, K., Sandgren, S., Mani, K., Belting, M., and Fransson, L.-A. (2001) J. Biol. Chem. 276, 46779-46791). Here, using both biochemical and microscopic techniques, we have identified and isolated S-nitrosylated forms of glypican-1 as well as slightly charged glypican-1 glycoforms containing heparan sulfate chains rich in N-unsubstituted glucosamines. These glycoforms were converted to highly charged species upon treatment of cells with 1 mm l-ascorbate, which releases NO from nitrosothiols, resulting in deaminative cleavage of heparan sulfate at the N unsubstituted glucosamines. S-Nitrosylation and subsequent deaminative cleavage were abrogated by inhibition of a Cu(2+)/Cu(+) redox cycle. Under cell-free conditions, purified S-nitrosylated glypican-1 was able to autocleave its heparan sulfate chains when NO release was triggered by l-ascorbate. The heparan sulfate fragments generated in cells during this autocatalytic process contained terminal anhydromannose residues. We conclude that the core protein of glypican-1 can slowly accumulate NO as nitrosothiols, whereas Cu(2+) is reduced to Cu(+). Subsequent release of NO results in efficient deaminative cleavage of the heparan sulfate chains attached to the same core protein, whereas Cu(+) is oxidized to Cu(2+). PMID- 12084717 TI - Secretory leucoprotease inhibitor prevents lipopolysaccharide-induced IkappaBalpha degradation without affecting phosphorylation or ubiquitination. AB - Secretory leucoprotease inhibitor (SLPI) is a non-glycosylated protein produced by epithelial cells, macrophages, and neutrophils and was initially identified as a serine protease inhibitor of the neutrophil proteases elastase and cathepsin G. In addition to its antiprotease activity, SLPI has been shown to exhibit anti inflammatory properties including down-regulation of tumor necrosis factor-alpha expression by lipopolysaccharide (LPS) in monocytes, inhibition of NF-kappaB activation by IgG immune complexes in a rat model of acute lung injury, and prevention of human immunodeficiency virus infectivity in monocytic cells via as yet unidentified mechanisms. In this report we have shown that SLPI prevents LPS induced NF-kappaB activation by inhibiting degradation of IkappaBalpha without affecting the LPS-induced phosphorylation and ubiquitination of IkappaBalpha. We have also demonstrated that SLPI prevents LPS-induced interleukin-1 receptor associated kinase and IkappaBbeta degradation. In addition, we have demonstrated that oxidized SLPI, a variant of SLPI that has diminished antiprotease activity, cannot prevent LPS-induced NF-kappaB activation or Inhibitor kappaB alpha/beta degradation indicating that the anti-inflammatory effect of SLPI on the LPS signaling pathway is dependent on its antiprotease activity. These results suggest that SLPI may be inhibiting proteasomal degradation of NF-kappaB regulatory proteins, an effect that is dependent on the antiprotease activity of SLPI. PMID- 12084718 TI - Regulation of hyaluronidase activity by alternative mRNA splicing. AB - Hyaluronidase is a hyaluronic acid-degrading endoglycosidase that is present in many toxins and the levels of which are elevated in cancer. Increased concentration of HYAL1-type hyaluronidase correlates with tumor progression and is a marker for grade (G) 2 or 3 bladder cancer. Using bladder tissues and cells, prostate cancer cells, and kidney tissues and performing reverse transcription PCR, cDNA cloning, DNA sequencing, and in vitro translation, we identified splice variants of HYAL1 and HYAL3. HYAL1v1 variant lacks a 30-amino acid (aa) sequence (301-330) present in HYAL1 protein. HYAL1v1, HYAL1v2 (aa 183-435 present in HYAL1 wild type), HYAL1v3 (aa 1-207), HYAL1v4 (aa 260-435), and HYAL1v5 (aa 340-435) are enzymatically inactive and are expressed in normal tissues/cells and G1 bladder tumor tissues. However, HYAL1 wild type is expressed in G2/G3 tumors and in invasive tumor cells. Stable transfection and HYAL1v1-specific antibody confirmed that the HYAL1 sequence from aa 301 to 330 is critical for hyaluronidase activity. All tumor cells and tissues mainly express HYAL3 variants. HYAL3v1 lacks a 30-aa sequence (299-328) present in HYAL3 protein, that is homologous to the 30-aa HYAL1 sequence. HYAL3v1, HYAL3v2 (aa 251-417 present in HYAL3 wild type), and HYAL3v3 (aa 251-417, but lacking aa 299-328), are enzymatically inactive. Although splicing of a single independent exon generates HYAL1v1 and HYAL3v1, internal exon splicing generates the other HYAL1/HYAL3 variants. These results demonstrate that alternative mRNA splicing controls cellular expression of enzymatically active hyaluronidase and may explain the elevated hyaluronidase levels in bladder/prostate cancer. PMID- 12084720 TI - Identification of human male germ cell-associated kinase, a kinase transcriptionally activated by androgen in prostate cancer cells. AB - Androgen is involved in both normal development and malignant transformation of prostate cells. The signal transduction pathways associated with these processes are not well understood. Using a novel kinase display approach, we have identified a protein kinase, human male germ cell-associated kinase (hMAK), which is transcriptionally induced by the androgenic hormone 5alpha-dihydrotestosterone (DHT). The kinetics of induction is rapid and dose-dependent, and the induction is not blocked by cycloheximide treatment. Real time reverse transcription-PCR studies demonstrated a 9-fold induction of hMAK by 10 nm DHT at 24 h post stimulation. The expression levels of hMAK in prostate cancer cell lines are in general higher than those of normal prostate epithelial cells. A reverse transcription-PCR product encompassing the entire hMAK open reading frame was isolated. The results from sequencing analysis showed that the hMAK protein is 623 amino acids in length and contains a kinase catalytic domain at its N terminus, followed by a proline/glutamine-rich domain. The catalytic domain of this kinase contains sequence motifs related to both the cyclin-dependent kinase and the mitogen-activated protein kinase families. When expressed in COS1 cells, hMAK is kinase-active as demonstrated by autophosphorylation and phosphorylation of exogenous substrate and is localized in the nucleus. A 3.7-kilobase pair promoter of the hMAK locus was isolated from a human genomic DNA bacterial artificial chromosome clone and was shown to be activated by DHT. This activation can be blocked by an anti-androgen drug bicalutamide (Casodex), implicating the involvement of androgen receptor in this process. Taken together, these data suggest that hMAK is a protein kinase targeted by androgen that may participate in androgen-mediated signaling in prostate cancer cells. PMID- 12084719 TI - Role for 18:1 lysophosphatidic acid as an autocrine mediator in prostate cancer cells. AB - Lysophosphatidic acid (LPA) is a lipid mediator that may play an important role in growth and survival of carcinomas. In this study, LPA production and response were characterized in two human prostate cancer (CaP) cell lines: PC-3 and Du145. Bombesin, a neuroendocrine peptide that is mitogenic for CaP cells, stimulated focal adhesion kinase phosphorylation and activated the extracellular signal regulated kinase/mitogen-activated protein kinase pathway. Similar responses were elicited by 18:1 LPA (oleoyl-LPA). Studies using radioisotopic labeling revealed that both PC-3 and Du145 generate LPA and that LPA production is increased by bombesin. The kinetics of bombesin-induced phospholipase D activation and LPA production were similar. Using electrospray ionization mass spectrometry, 18:1 LPA was found to be an abundant LPA species in CaP cell medium. Structure activity studies of acyl-LPAs revealed that 18:1 LPA is most efficacious for activation of extracellular signal-regulated kinase and phospholipase D in CaP cells. Incubation with 18:1 LPA caused homologous desensitization of LPA response, whereas bombesin caused heterologous desensitization. LPA was present at nanomolar levels in medium from bombesin-treated cells. LPA extracted from the medium induced calcium mobilization in CaP cells. These results demonstrate that bioactive LPA is generated by CaP cells in response to a mitogen and suggest that 18:1 LPA can act as an autocrine mediator. PMID- 12084721 TI - FGF3 attached to a phosholipid membrane anchor gains a high transforming capacity. Implications of microdomains for FGF3 cell transformation. AB - NIH3T3 cells transformed by mouse FGF3-cDNA (DMI cells) selected for their ability to grow as anchorage-independent colonies in soft agar and in defined medium lacking growth factors exhibit a highly transformed phenotype. We have used dominant negative (DN) fibroblast growth factor (FGF) receptor 2 (FGFR2) isoforms to block the FGF response in DMI cells. When the DN-FGFR was expressed in DMI cells, their transformed phenotype can be reverted. The truncated FGFR2(IIIb), the high affinity FGFR for FGF3, is significantly more efficient at reverting the transformed phenotype as the IIIc isoform, reaffirming the notion that the affinity of the ligand to the DN-FGFR2 isoform determines the effect. Heparin or heparan sulfate displaces FGF3 from binding sites on the cell surface inhibiting the growth of DMI cells and reverts the transformed phenotype (). However, the presence of heparin is necessary to induce a mitogenic response in NIH3T3 cells when stimulated with soluble purified mouse FGF3. We have investigated the importance of cell surface binding of FGF3 for its ability to transform NIH3T3 cells by creating an FGF3 mutant anchored to the membrane via glycosylphosphatidylinositol (GPI). The GPI anchor renders the cell surface association of FGF3 independent from binding to heparan sulfate-proteoglycan of the cell surface membrane. Attachment of a GPI anchor to FGF3 also confers a much higher transforming potential to the growth factor. Even more, the purified GPI attached FGF3 is as much transforming as the secreted protein acting in an autocrine mode. Because NIH3T3 cells do not express the high affinity tyrosine kinase FGF receptors for FGF3, these findings suggest that FGF3 attached to GPI linked heparan sulfate-proteoglycan may have a broader biological activity as when bound to transmembrane or soluble heparan sulfate-proteoglycan. PMID- 12084722 TI - Naturally occurring mutations in the largest extracellular loops of ABCA1 can disrupt its direct interaction with apolipoprotein A-I. AB - The ABCA1 transporter contains two large domains into which many of the genetic mutations in individuals with Tangier disease fall. To investigate the structural requirements for the cellular cholesterol efflux mediated by ABCA1, we have determined the topology of these two domains and generated transporters harboring five naturally occurring missense mutations in them. These mutants, unlike wild type ABCA1, produced little or no apoA-I-stimulated cholesterol efflux when transfected into 293 cells, establishing their causality in Tangier disease. Because all five mutant proteins were well expressed and detectable on the plasma membrane, their interaction with the ABCA1 ligand, apolipoprotein (apo) A-I, was measured using bifunctional cross-linking agents. Four of five mutants had a marked decline in cross-linking to apoA-I, whereas one (W590S) retained full cross-linking activity. Cross-linking of apoA-I was temperature-dependent, rapid in onset, and detectable with both lipid- and water-soluble cross-linking agents. These results suggest that apoA-I-stimulated cholesterol efflux cannot occur without a direct interaction between the apoprotein and critical residues in two extracellular loops of ABCA1. The behavior of the W590S mutant indicates that although binding of apoA-I by ABCA1 may be necessary, it is not sufficient for stimulation of cholesterol efflux. PMID- 12084723 TI - An osmotically induced cytosolic Ca2+ transient activates calcineurin signaling to mediate ion homeostasis and salt tolerance of Saccharomyces cerevisiae. AB - Hyperosmotic stress caused by NaCl, LiCl, or sorbitol induces an immediate and short duration ( approximately 1 min) transient cytosolic Ca(2+) ([Ca(2+)](cyt)) increase (Ca(2+)-dependent aequorin luminescence) in Saccharomyces cerevisiae cells. The amplitude of the osmotically induced [Ca(2+)](cyt) transient was attenuated by the addition of chelating agents EGTA or BAPTA, cation channel pore blockers, competitive inhibitors of Ca(2+) transport, or mutations (cch1Delta or mid1Delta) that reduce Ca(2+) influx, indicating that Ca(ext)(2+) is a source for the transient. An osmotic pretreatment (30 min) administered by inoculating cells into media supplemented with either NaCl (0.4 or 0.5 m) or sorbitol (0.8 or 1.0 m) enhanced the subsequent growth of these cells in media containing 1 m NaCl or 2 m sorbitol. Inclusion of EGTA in the osmotic pretreatment media or the cch1Delta mutation reduced cellular capacity for NaCl but not hyperosmotic adaptation. The stress-adaptive effect of hyperosmotic pretreatment was mimicked by exposing cells briefly to 20 mm CaCl(2). Thus, NaCl- or sorbitol-induced hyperosmotic shock causes a [Ca(2+)](cyt) transient that is facilitated by Ca(2+) influx, which enhances ionic but not osmotic stress adaptation. NaCl-induced ENA1 expression was inhibited by EGTA, cch1Delta mutation, and FK506, indicating that the [Ca(2+)](cyt) transient activates calcineurin signaling to mediate ion homeostasis and salt tolerance. PMID- 12084724 TI - Requirement of BMP-2-induced phosphatidylinositol 3-kinase and Akt serine/threonine kinase in osteoblast differentiation and Smad-dependent BMP-2 gene transcription. AB - The mechanism by which bone morphogenetic protein-2 (BMP-2) induces osteoblast differentiation is not precisely known. We investigated the involvement of the phosphatidylinositol (PI) 3-kinase/Akt signal transduction pathway in modulation of this process. BMP-2 stimulated PI 3-kinase activity in osteogenic cells. Inhibition of PI 3-kinase activity with the specific inhibitor Ly-294002 prevented BMP-2-induced alkaline phosphatase, an early marker of osteoblast differentiation. Expression of dominant-negative PI 3-kinase also abolished osteoblastic induction of alkaline phosphatase in response to BMP-2, confirming the involvement of this lipid kinase in this process. BMP-2 stimulated Akt serine/threonine kinase activity in a PI 3-kinase-dependent manner in osteoblast precursor cells. Inhibition of Akt activity by a dominant-negative mutant of Akt blocked BMP-2-induced osteoblastic alkaline phosphatase activity. BMP-2 stimulates its own expression during osteoblast differentiation. Expression of dominant-negative PI 3-kinase or dominant-negative Akt inhibited BMP-2-induced BMP-2 transcription. Because all the known biological activities of BMP-2 are mediated by transcription via BMP-specific Smad proteins, we investigated the involvement of PI 3-kinase in Smad-dependent BMP-2 transcription. Smad5 stimulated BMP-2 transcription independent of addition of the ligand. Dominant negative PI 3-kinase or dominant-negative Akt inhibited Smad5-dependent transcription of BMP-2. Furthermore dominant-negative Akt inhibited translocation of BMP-specific Smads into nucleus. Together these data provide the first evidence that activation of BMP receptor serine/threonine kinase stimulates the PI 3 kinase/Akt pathway and define a role for this signal transduction pathway in BMP-specific Smad function during osteoblast differentiation. PMID- 12084725 TI - Transcriptional activation of the human leptin gene in response to hypoxia. Involvement of hypoxia-inducible factor 1. AB - In addition to having a major role in energy homeostasis, leptin is emerging as a pleiotropic cytokine with multiple physiological effector functions. The recently discovered proangiogenic activity of leptin suggested the hypothesis that its production might be regulated by hypoxia, as are other angiogenic factors. To examine this proposal, the expression of leptin protein and mRNA was measured and found to be markedly up-regulated in response to ambient or chemical hypoxia (upon exposure to desferrioxamine or cobalt chloride), an effect that requires intact RNA synthesis, suggesting a transcriptional mechanism. Transient transfection of cultured cells with deletion constructs of the leptin gene promoter linked to a reporter gene revealed a functional hypoxia response element (HRE) located at position -116 within the proximal upstream region. This putative HRE harbors a characteristic 5'-RCGTG-3' core motif, a hallmark of hypoxia sensitive genes and recognized by the hypoxia-inducible factor 1 (HIF1), which consists of a HIF1alpha/HIFbeta heterodimer. Constructs harboring this -116/HRE supported reporter gene expression in response to hypoxia but not when mutated. Expression of HIF1alpha cDNA in normoxic cells mimicked hypoxia-induced reporter gene expression in cells cotransfected with the wild type leptin -116/HRE construct but not with the mutant. Gel shift assays with a (32)P-labeled leptin promoter -116/HRE probe and nuclear extracts from hypoxia-treated cells indicated binding of the HIF1alpha/beta heterodimer, which was blocked with an excess of unlabeled -116/HRE probe or a HIF1-binding probe from the erythropoietin gene enhancer. Taken together, these observations demonstrate that the leptin gene is actively engaged by hypoxia through a transcriptional pathway commonly utilized by hypoxia-sensitive genes. PMID- 12084726 TI - Targeted disruption of Np95 gene renders murine embryonic stem cells hypersensitive to DNA damaging agents and DNA replication blocks. AB - NP95, which contains a ubiquitin-like domain, a cyclin A/E-Cdk2 phosphorylation site, a retinoblastoma (Rb) binding motif, and a ring finger domain, has been shown to be colocalized as foci with proliferating cell nuclear antigen in early and mid-S phase nuclei. We established Np95 nulligous embryonic stem cells by replacing the exons 2-7 of the Np95 gene with a neo cassette and by selecting out a spontaneously occurring homologous chromosome crossing over with a higher concentration of neomycin. Np95-null cells were more sensitive to x-rays, UV light, N-methyl-N"-nitro-N-nitrosoguanidine (MNNG), and hydroxyurea than embryonic stem wild type (Np95(+/+)) or heterozygously inactivated (Np95(+/-)) cells. Expression of transfected Np95 cDNA in Np95-null cells restored the resistance to x-rays, UV, MNNG, or hydroxyurea concurrently to a level similar to that of Np95(+/-) cells, although slightly below that of wild type (Np95(+/+)) cells. These findings suggest that NP95 plays a role in the repair of DNA damage incurred by these agents. The frequency of spontaneous sister chromatid exchange was significantly higher for Np95-null cells than for Np95(+/+) cells or Np95(+/ ) cells (p < 0.001). We conclude that NP95 functions as a common component in the multiple response pathways against DNA damage and replication arrest and thereby contributes to genomic stability. PMID- 12084727 TI - A novel biosynthetic pathway providing precursors for fatty acid biosynthesis and secondary metabolite formation in myxobacteria. AB - Short chain carboxylic acids are well known as the precursors of fatty acid and polyketide biosynthesis. Iso-fatty acids, which are important for the control of membrane fluidity, are formed from branched chain starter units (isovaleryl-CoA and isobutyryl-CoA), which in turn are derived from the degradation of leucine and valine, respectively. Branched chain carboxylic acids are also employed as starter molecules for the biosynthesis of secondary metabolites, e.g. the therapeutically important anthelmintic agent avermectin or the electron transport inhibitor myxothiazol. During our studies on myxothiazol biosynthesis in the myxobacterium, Stigmatella aurantiaca, we detected a novel biosynthetic route to isovaleric acid. After cloning and inactivation of the branched chain keto acid dehydrogenase complex, which is responsible for the degradation of branched chain amino acids, the strain is still able to produce iso-fatty acids and myxothiazol. Incorporation studies employing deuterated leucine show that it can only serve as precursor in the wild type strain but not in the esg mutant. Feeding experiments using (13)C-labeled precursors show that isovalerate is efficiently made from acetate, giving rise to a labeling pattern in myxothiazol that provides evidence for a novel branch of the mevalonate pathway involving the intermediate 3,3 dimethylacryloyl-CoA. 3,3-Dimethylacrylic acid was synthesized in deuterated form and fed to the esg mutant, resulting in strong incorporation into myxothiazol and iso-fatty acids. Similar experiments employing Myxococcus xanthus revealed that the discovered biosynthetic route described is present in other myxobacteria as well. PMID- 12084728 TI - Aggregation of misfolded proteins can be a selective process dependent upon peptide composition. AB - Intracellular aggregation of misfolded proteins is observed in a number of human diseases, in particular, neurologic disorders in which expanded tracts of polyglutamine residues play a central role. A variety of other proteins are prone to aggregation when mutated, indicating that this process is a common pathologic mechanism for inherited disorders. However, little is known about the relationship between the sequence of aggregating peptides and the specificity of intracellular accumulation. Here we demonstrate that substitution of two residues eliminates aggregation of a 111-amino acid peptide derived from the C-terminal portion of the cystic fibrosis transmembrane conductance regulator (CFTR). We also show that fusion to a reporter protein considerably alters the subcellular distribution of aggregating peptide. When fused to green fluorescent protein, the peptide containing amino acids 1370-1480 of CFTR accumulates in large perinuclear or nuclear aggregates. The same CFTR fragment devoid of green fluorescent protein localizes predominantly to discrete accumulations associated with mitochondria. Importantly, both types of accumulation are dependent on the presence of the same two amino acids within the CFTR sequence. Co-expression studies show that both CFTR-derived proteins can co-localize in large cytoplasmic/nuclear aggregates. However, neither CFTR construct accumulates in intracellular inclusions formed by N-terminal fragment of huntingtin. In addition to unique accumulation patterns, each aggregating peptide shows differences in association with chaperone proteins. Thus, our results indicate that the process of intracellular aggregation can be a selective process determined by the composition of the aggregating peptides. PMID- 12084729 TI - Comparison of Cak1p-like cyclin-dependent kinase-activating kinases. AB - Cyclin-dependent kinases (cdks) coordinate progression through the eukaryotic cell cycle and require phosphorylation by a cdk-activating kinase (CAK) for full activity. In most eukaryotes Cdk7 is the catalytic subunit of a heterotrimeric CAK (Cdk7-cyclin H-Mat1) that is also involved in transcription as part of the transcription factor IIH complex. The Saccharomyces cerevisiae CAK, Cak1p, is a monomeric protein kinase with an atypical sequence and unusual biochemical properties compared with trimeric CAKs and other protein kinases. We sought to determine whether these properties were shared by a small group of monomeric CAKs that can function in place of CAK1 in S. cerevisiae. We found that Schizosaccharomyces pombe Csk1, Candida albicans Cak1, and Arabidopsis thaliana Cak1At, like Cak1p, all displayed a preference for cyclin-free cdk substrates, were insensitive to the protein kinase inhibitor 5' fluorosulfonylbenzoyladenosine (FSBA), and were insensitive to mutation of a highly conserved lysine residue found in the nucleotide binding pocket of all protein kinases. The S. pombe and C. albicans kinases also resembled Cak1p in their kinetics of nucleotide and protein substrate utilization. Conservation of these unusual properties in fungi and plants points to shared evolutionary requirements not met by Cdk7 and raises the possibility of developing antifungal agents targeting CAKs. PMID- 12084730 TI - The synthesis of inositol hexakisphosphate. Characterization of human inositol 1,3,4,5,6-pentakisphosphate 2-kinase. AB - The enzyme(s) responsible for the production of inositol hexakisphosphate (InsP(6)) in vertebrate cells are unknown. In fungal cells, a 2-kinase designated Ipk1 is responsible for synthesis of InsP(6) by phosphorylation of inositol 1,3,4,5,6-pentakisphosphate (InsP(5)). Based on limited conserved sequence motifs among five Ipk1 proteins from different fungal species, we have identified a human genomic DNA sequence on chromosome 9 that encodes human inositol 1,3,4,5,6 pentakisphosphate 2-kinase (InsP(5) 2-kinase). Recombinant human enzyme was produced in Sf21 cells, purified, and shown to catalyze the synthesis of InsP(6) or phytic acid in vitro. The recombinant protein converted 31 nmol of InsP(5) to InsP(6)/min/mg of protein (V(max)). The Michaelis-Menten constant for InsP(5) was 0.4 microM and for ATP was 21 microM. Saccharomyces cerevisiae lacking IPK1 do not produce InsP(6) and show lethality in combination with a gle1 mutant allele. Here we show that expression of the human InsP(5) 2-kinase in a yeast ipk1 null strain restored the synthesis of InsP(6) and rescued the gle1-2 ipk1-4 lethal phenotype. Northern analysis on human tissues showed expression of the human InsP(5) 2-kinase mRNA predominantly in brain, heart, placenta, and testis. The isolation of the gene responsible for InsP(6) synthesis in mammalian cells will allow for further studies of the InsP(6) signaling functions. PMID- 12084732 TI - Retinal nerve fibre layer loss in diabetes mellitus without retinopathy. PMID- 12084733 TI - Surgery for glaucoma in the 21st century. PMID- 12084734 TI - Antique ophthalmic instruments and books: the Royal College Museum. PMID- 12084736 TI - Models for improving cataract surgical rates in southern China. AB - BACKGROUND/AIMS: Unoperated cataract is the main challenge to blindness reduction in China. This demonstration project in Guangdong Province was designed to test various strategies to improve the cataract surgical rate. METHODS: Two strategies (reduction of the cataract surgical fee and training of country doctors in the detection of cataract blindness) were implemented in each of two counties. Both interventions were introduced in a third county. RESULTS: The cataract surgery rate (CSR) per million was 366 in the county where training took place, 588 where the fee was lowered, and 1140 where both interventions took place. The improvement in CSR was highly significant (p<0.001). CONCLUSIONS: In southern China, CSR can be increased with community based measures. Sustainability for such measures will be the major future challenge. PMID- 12084735 TI - Visual impairment and blindness in Europe and their prevention. AB - The European region currently differs in many aspects, such as political, socioeconomic, and geographical. After substantial political changes at the beginning of the 1990s, the majority of central and eastern European countries started to rebuild their healthcare systems. It is apparent that eastern Europe represents a highly diverse region where the difference among countries broadens year after year. In highly industrialised countries of Europe, the leading causes of childhood serious visual loss are lesions of the central nervous system, congenital anomalies and retinal disorders. In the middle income countries of Europe, congenital cataract, glaucoma and, mainly, retinopathy of prematurity are highly expressed. The major cause of serious visual loss in adults in industrialised countries is age related macular degeneration. The other conditions comprise cataract, glaucoma, diabetic retinopathy, and uncorrected/uncorrectable refractive errors, along with low vision. In people of working age, diabetic retinopathy, retinopathy pigmentosa, and optic atrophy are the most frequently reported causes of serious visual loss. In the middle income countries of Europe, advanced cataract, glaucoma, and diabetic retinopathy are more frequently observed. PMID- 12084737 TI - Retinal nerve fibre layer loss in patients with type 1 diabetes mellitus without retinopathy. AB - BACKGROUND/AIM: There is evidence suggesting the occurrence of neurovisual abnormalities in patients with diabetes without retinopathy. However, the determination of abnormalities in the neural and glial elements in vivo is difficult. The aim of this study was to investigate whether a retinal nerve fibre layer (RNFL) defect (as determined by scanning laser polarimetry, SLP) is present in patients without clinical manifestations of diabetic retinopathy. METHODS: 12 patients with type 1 diabetes mellitus (DM) without retinopathy or other diabetes induced microvascular complications, underwent a complete ophthalmological examination, including automated perimetry and RNFL measurements with a nerve fibre layer analyser GDx. The data were compared with a normal control group matched for age and sex. RESULTS: The superior segment retardation in patients with diabetes was lower than in the control group, based on the superior integral (0.19 (SD 0.06) v 0.23 (0.04) mm(2), p=0.03) and the superior average (71.0 (11.05) v 84.27 (10.56) microm, p=0.007) parameters. CONCLUSION: This finding may be indicative of significant nerve fibre loss in the superior segment of the retina in patients with type 1 diabetes mellitus but without retinopathy. The meaning of intraretinal differences in RNFL retardation, indicating asymmetric NFL loss, in patients with diabetes is yet not understood. PMID- 12084738 TI - A novel mutation in KRT12 associated with Meesmann's epithelial corneal dystrophy. AB - BACKGROUND: The molecular basis of Meesmann's epithelial corneal dystrophy (MECD) has recently been attributed to mutations in the cornea specific keratin genes KRT3 and KRT12. The mechanisms by which these mutations cause the Meesmann's phenotype are not clear. This study presents new data, examines clinical, histological, ultrastructural, and molecular aspects of MECD, and compares the features seen in this condition with those observed in other well studied keratin diseases such as epidermolysis bullosa simplex. METHODS: A two generation family with typical features of Meesmann's epithelial corneal dystrophy (MECD) was studied. All family members were examined under a slit lamp. Biopsy material from elective keratoplasty was studied by histopathological and ultrastructural analysis using standard techniques. Direct automated sequencing of genomic DNA was used for mutation detection, mutations were confirmed by restriction digest analysis. RESULTS: The abnormal corneal epithelium was acanthotic and contained numerous dyskeratotic cells and intraepithelial vesicles. By electron microscopy abnormally aggregated and clumped keratin filament bundles were detected in basal and suprabasal keratinocytes from the centre of the cornea. Direct sequencing of the patients' genomic DNA revealed a novel missense mutation (423T>G) in exon 1 of the cornea specific keratin 12 (KRT12) gene. This mutation predicts the amino acid change N133K within the helix initiation motif of the K12 polypeptide. Comparative studies with well established keratin disorders of other human epithelia underscore the pathogenic relevance of K3 and K12 gene mutations in Meesmann's epithelial corneal dystrophy. The morphological data presented here illustrate the disruptive effects of keratin gene mutations on the integrity of corneal keratinocytes. CONCLUSIONS: A clinical, histopathological, and ultrastructural study of a previously unreported family with MECD is presented. In this family the disease is ascribed to a novel mutation in KRT12. A molecular mechanism is proposed for MECD based on the comparison with other well characterised keratin diseases. PMID- 12084739 TI - Role of argon laser trabeculoplasty as primary and secondary therapy in open angle glaucoma in Indian patients. AB - AIMS: To study the efficacy of argon laser trabeculoplasty (ALT) as a primary and secondary therapy in Indian patients with primary open angle glaucoma (POAG). METHODS: ALT was performed as a primary therapy in 40 eyes of 21 patients newly diagnosed with POAG (group I) and as a secondary therapy in 39 eyes of 21 patients who had been taking topical antiglaucoma medications for more than 1 year. The best corrected visual acuity, baseline intraocular pressure (IOP), diurnal variation of IOP, anterior chamber angles, and visual fields were the various parameters evaluated before and after performing ALT. Follow up visits were scheduled at 24 hours, 1 week, 1 month, 3 months, 6 months, and 1 year after the surgery. The patients were recalled at the end of 5 years for the final follow up examination. Success of ALT was defined as IOP 15% in twice as many males than expected (62% v 28%, p <0.0001). This rose to almost six times when cCHDRs greater than 30% were compared (17% v 3%, p = 0.002). There was a fourfold increase in the proportion of female RVO patients with a cCHDR above 15% (40% v 9%, p <0.0001) and at a cCHDR of 30% and above (10% v 0%, p = 0.004). There were also significant differences in the cCHDR between central and branch RVO (both sexes). The branch form of RVO (BRVO) having higher cCHDRs because of systolic hypertension (164.1 (21.6) mm Hg v 149.5 (23.5) mm Hg, p = 0.003) and age (61.7 (8.3) years v 56.7 (10.6) years, p = 0.017). CONCLUSIONS: RVO is the presenting complaint in a group of patients at increased risk of CVD and is in agreement with the long term follow up data demonstrating an increased mortality from CVD in patients with RVO. The Framingham algorithm can accurately determine the cCHDR (or cCVDR) to assist the clinician in deciding who to treat in accordance with the Joint British Societies' guidelines, with particular regard to hypertension, lipid lowering, and the use of aspirin therapy. PMID- 12084749 TI - Perceived quality of health care in macular disease: a survey of members of the Macular Disease Society. AB - AIM: To investigate the experiences of people with macular disease within the British healthcare system. METHOD: The Macular Disease Society Questionnaire, a self completion questionnaire designed to survey the experiences of people with macular disease, was sent to 2000 randomly selected members of the Macular Disease Society. The questionnaire incorporated items about people's experiences with health professionals and the information and support provided by them at the time of diagnosis and thereafter. RESULTS: Over 50% thought their consultant eye specialist was not interested in them as a person and 40% were dissatisfied with their diagnostic consultation. 185 people thought their general practitioner (GP) was well informed about macular disease but twice as many people thought their GP was not well informed. About an equal number of people thought their GP was supportive as those who thought their GP was not supportive. A total of 1247 people were told "nothing can be done to help with your macular disease." A number of negative emotional reactions were experienced by those people as a result, with 61% of them reporting feeling anxious or depressed. Of 282 people experiencing visual hallucinations after diagnosis with macular disease, only 20.9% were offered explanations for them. CONCLUSIONS: Many people with macular disease have unsatisfactory experiences of the healthcare system. Many of the reasons for dissatisfaction could be resolved by healthcare professionals if they were better informed about macular disease and had a better understanding of and empathy with patients' experiences. PMID- 12084750 TI - Aetiology of congenital and paediatric cataract in an Australian population. AB - BACKGROUND/AIM: Paediatric cataract is a major cause of childhood blindness. Several genes associated with congenital and paediatric cataracts have been identified. The aim was to determine the incidence of cataract in a population, the proportion of hereditary cataracts, the mode of inheritance, and the clinical presentation. METHODS: The Royal Children's Hospital and the Royal Victorian Eye and Ear Hospital have a referral base for almost all paediatric patients with cataracts in south eastern Australia. The database contains cases seen over the past 25 years. The medical histories of these patients were reviewed. RESULTS: 421 patients with paediatric cataract were identified, which gives an estimated incidence of 2.2 per 10,000 births. Of the 342 affected individuals with a negative family history, 50% were diagnosed during the first year of life, and 56/342 (16%) were associated with a recognised systemic disease or syndrome. Unilateral cataract was identified in 178/342 (52%) of sporadic cases. 79 children (from 54 nuclear families) had a positive family history. Of these 54 families, 45 were recruited for clinical examination and DNA collection. Ten nuclear families were subsequently found to be related, resulting in four larger pedigrees. Thus, 39 families have been studied. The mode of inheritance was autosomal dominant in 30 families, X linked in four, autosomal recessive in two, and uncertain in three. In total, 178 affected family members were examined; of these 8% presented with unilateral cataracts and 43% were diagnosed within the first year of life. CONCLUSIONS: In the paediatric cataract population examined, approximately half of the patients were diagnosed in the first year of life. More than 18% had a positive family history of cataracts. Of patients with hereditary cataracts 8% presented with unilateral involvement. Identification of the genes that cause paediatric and congenital cataract should help clarify the aetiology of some sporadic and unilateral cataracts. PMID- 12084751 TI - A randomised controlled trial of written information: the effect on parental non concordance with occlusion therapy. AB - BACKGROUND/AIMS: It is well documented that non-concordance with occlusion therapy is both substantial and a major factor leading to treatment failure. Parental understanding in previous work has been found to be poor in key areas such as the critical period and effect of age on prognosis. Research in other areas of medicine has shown that the level of understanding can have a direct effect on the level of concordance. The aims of this study were to assess the ability of educational material in the form of a leaflet, to improve parental understanding of amblyopia and occlusion, and subsequently increase concordance. METHODS: Parents of children aged between 1 and 7 years receiving a minimum of 1 hour of occlusion for amblyopia were recruited. A randomised controlled trial was undertaken where, on inclusion, the patients were randomised into a leaflet group, whose parents were issued with written educational material, and a control group whose parents did not receive the written information. Patients were paired and matched for age (<2 years difference) and amount of prescribed occlusion (no more than 1 hour difference). Concordance was monitored by a parental diary and knowledge and parental reasons for non-concordance were assessed by a questionnaire. Concordance was analysed by means of a concordance index and by calculating the proportion of non-concordant parents by setting a threshold of concordance at 80%. RESULTS: Parental knowledge was significantly greater in the leaflet group (88% had complete knowledge) compared to the control group (49% had complete knowledge) (p <0.001). There were also differences between the groups in the area of the treatment regimen, with errors only occurring in the control group (three patients occluded the incorrect eye), but this did not reach statistical significance. Concordance was significantly greater in the leaflet group (mean concordance index 0.85) compared to the non-leaflet group (mean concordance index (0.71) (p <0.001). Comparison of the proportion of non concordant parents was also statistically different (p <0.005) at 0.23 (95% CI 0.13 to 0.35) for the leaflet group compared to 0.54 (95% CI 0.41 to 0.67) for the control group. CONCLUSION: A large proportion of patients would benefit by increasing parental knowledge in key areas such as the critical period, importance of occlusion, and potential negative consequences of not treating amblyopia. Written information is a simple, inexpensive, easy to implement, yet effective method of improving parental understanding and subsequent concordance. PMID- 12084752 TI - Analysis of the results of surgical endoscopic dacryocystorhinostomy: effect of the level of obstruction. AB - AIM: One of the main factors in determining success rate of lacrimal surgery is the level of obstruction in the lacrimal drainage system. There are only few reports which quantify this, and none on endoscopic dacryocystorhinostomy (DCR). METHODS: A case series of patients who had endoscopic DCR for anatomical obstruction of the lacrimal drainage system was performed. All patients who had lacrimal blockage referred to a district general hospital, irrespective of the level of blockage, had endoscopic DCR as the initial treatment by the authors. A total of 191 endoscopic DCRs were performed between 1994 and 1999. No other forms of lacrimal surgery were performed during this period. The level of the obstruction was assessed by the ophthalmologist before the operation and confirmed at surgery. All cases were followed up for a minimum of 6 months, and 96 cases were also reviewed 12 months after surgery. The outcome of the endoscopic DCR operation for each eye was categorised into complete cure, partial cure, or no improvement according to the degree of symptomatic relief following the operation. RESULTS: Complete relief from epiphora was achieved in 89% of cases overall at 6 months. The success rate in cases with lacrimal sac/duct obstruction (93%) or common canalicular blockage (88%) was comparable. In canalicular obstruction, however, the complete cure rate was lower at 54%. The benefit of the operation was maintained at 12 months. CONCLUSION: This study demonstrates that the success rate of surgical (non-laser) endoscopic DCR is comparable to that reported for external DCR. Moreover, the technique is appropriate for initial treatment of patients with common canalicular or even canalicular obstruction. PMID- 12084753 TI - Prevalence of visual impairment in people aged 75 years and older in Britain: results from the MRC trial of assessment and management of older people in the community. AB - AIMS: To measure the prevalence of visual impairment in a large representative sample of people aged 75 years and over participating in the MRC trial of assessment and management of older people in the community. METHODS: 53 practices in the MRC general practice research framework. Data were obtained from 14 600 participants aged 75 years and older. Prevalence of visual impairment overall (binocular visual acuity <6/18) which was categorised separately into low vision (binocular visual acuity <6/18-3/60) or blindness (binocular visual acuity of <3/60). The prevalence of binocular acuity <6/12 was presented for comparison with other studies. Visual acuity was measured using Glasgow acuity charts; glasses, if worn, were not removed. RESULTS: Visual acuity was available for 14 600 people out of 21 241 invited (69%). Among people with visual acuity data, 12.4% overall (1803) were visually impaired (95% confidence intervals 10.8% to 13.9%); 1501 (10.3%) were categorised as having low vision (8.7% to 11.8%), and 302 (2.1%) were blind (1.8% to 2.4%). At ages 75-79, 6.2% of the cohort were visually impaired (5.1% to 7.3%) with 36.9% at age 90+ (32.5% to 41.3%). At ages 75-79, 0.6% (0.4% to 0.8%) of the study population were blind, with 6.9% (4.8% to 9.0%) at age 90+. In multivariate regression, controlling for age, women had significant excess risk of visual impairment (odds ratio 1.43, 95% confidence interval 1.29 to 1.58). Overall, 19.9% of study participants had a binocular acuity of less than 6/12 (17.8% to 22.0%). CONCLUSION: The results from this large study show that visual impairment is common in the older population and that this risk increases rapidly with advancing age, especially for women. A relatively conservative measure of visual impairment was used. If visual impairment had been defined as visual acuity of <6/12 (American definition of visual impairment), the age specific prevalence estimates would have increased by 60%. PMID- 12084754 TI - Automated tri-image analysis of stored corneal endothelium. AB - BACKGROUND: Endothelial examination of organ culture stored corneas is usually done manually and on several mosaic zones. Some banks use an image analyser that takes account of only one zone. This method is restricted by image quality, and may be inaccurate if endothelial cell density (ECD) within the mosaic is not homogeneous. The authors have developed an analyser that has tools for automatic error detection and correction, and can measure ECD and perform morphometry on multiple zones of three images of the endothelial mosaic. METHODS: 60 human corneas were divided into two equal groups: group 1 with homogeneous mosaics, group 2 with heterogeneous ones. Three standard microscopy video images of the endothelium, graded by quality, were analysed either in isolation (so called mono image analysis) or simultaneously (so called tri-image analysis), with 50 or 300 endothelial cells (ECs) counted. The automated analysis was compared with the manual analysis, which concerned 10 non-adjacent zones and about 300 cells. For each analysis method, failures and durations were studied according to image quality. RESULTS: All corneas were able to undergo analysis, in about 2 or 7.5 minutes for 50 and 300 ECs respectively. The tri-image analysis did not increase analysis time and never failed, even with mediocre images. The tri-image analysis of 300 ECs was always most highly correlated with the manual count, particularly in the heterogeneous cornea group (r=0.94, p<0.001) and prevented serious count errors. CONCLUSIONS: This analyser allows reliable and rapid analysis of ECD, even for heterogeneous endothelia mosaics and mediocre images. PMID- 12084755 TI - Comparative experiments for in vivo fibroplasia and biological stability of four porous polymers intended for use in the Seoul-type keratoprosthesis. AB - AIMS: To evaluate in vivo fibroplasia and biological stability of porous polymers intended for use in the Seoul-type keratoprosthesis (S-KPro). METHODS: Four porous polymers (polypropylene, two kinds of polyethylene terephthalate (PE70 and PE50), and polyurethane) were investigated. Discs of polymers were inserted into the corneal stroma of rabbits for a 2 and 5 month period. Corneal oedema and neovascularisation were evaluated. The fibroplasia and collagen deposition were examined under light and transmission electron microscopy. S-KPros, whose skirt was made of four types of polymer, were implanted into the rabbits' eyes. The retention time and complications were evaluated. RESULTS: Neovascularisation and corneal oedema were found in all of the disc inserted eyes, but the corneal oedema subsided within 2 months in most of the eyes. The mean number of fibroblasts increased significantly in polypropylene and PE50 disc inserted eyes compared with polyurethane disc inserted eyes. Plentiful collagen deposition was also found in both polypropylene and PE50 disc inserted eyes. Mean retention time in the polypropylene SK-Pro implanted eyes was longer than that of the other eyes (20.7 weeks). The PE70 skirt induced corneal melting around the prosthesis. CONCLUSION: Polypropylene encourages fibroblast ingrowth and shows good biological stability when used as a skirt material in S-KPro. PMID- 12084756 TI - The surgical correction of moderate hypermetropia: the management controversy. PMID- 12084758 TI - Histopathological findings in filtering blebs with recurrent blebitis. PMID- 12084757 TI - Glaucoma: squaring the psychophysics and neurobiology. AB - Advances in our understanding of the pathophysiology of retinal ganglion cell death in glaucoma are providing important insights into the functional changes occurring in retinal ganglion cells in the early stages of the disease. These exciting new findings may help us develop psychophysical tests to monitor early retinal ganglion cell damage, possibly before neurons are committed to the process of cell death. PMID- 12084759 TI - Retinal detachment surgery in district general hospitals: an audit of changing practice. PMID- 12084761 TI - Do climatic variables influence the development of posterior vitreous detachment? PMID- 12084760 TI - Ocular and systemic posaconazole(SCH-56592) treatment of invasive Fusarium solani keratitis and endophthalmitis. PMID- 12084762 TI - World Sight Day and cataract blindness. PMID- 12084763 TI - "No-needle" sub-Tenon's anaesthesia. PMID- 12084764 TI - Trachoma and recurrent trichiasis. PMID- 12084765 TI - Failure of amniotic membrane transplantation in the treatment of acute ocular burns. PMID- 12084766 TI - A re-analysis of astigmatism correction. PMID- 12084767 TI - Role of autologous serum in persistent epithelial defects. PMID- 12084770 TI - Current-dependent block of rabbit sino-atrial node I(f) channels by ivabradine. AB - "Funny" (f-) channels have a key role in generation of spontaneous activity of pacemaker cells and mediate autonomic control of cardiac rate; f-channels and the related neuronal h-channels are composed of hyperpolarization-activated, cyclic nucleotide-gated (HCN) channel subunits. We have investigated the block of f channels of rabbit cardiac sino-atrial node cells by ivabradine, a novel heart rate-reducing agent. Ivabradine is an open-channel blocker; however, block is exerted preferentially when channels deactivate on depolarization, and is relieved by long hyperpolarizing steps. These features give rise to use-dependent behavior. In this, the action of ivabradine on f-channels is similar to that reported of other rate-reducing agents such as UL-FS49 and ZD7288. However, other features of ivabradine-induced block are peculiar and do not comply with the hypothesis that the voltage-dependence of block is entirely attributable to either the sensitivity of ivabradine-charged molecules to the electrical field in the channel pore, or to differential affinity to different channel states, as has been proposed for UL-FS49 (DiFrancesco, D. 1994. Pflugers Arch. 427:64-70) and ZD7288 (Shin, S.K., B.S. Rotheberg, and G. Yellen. 2001. J. Gen. Physiol. 117:91 101), respectively. Experiments where current flows through channels is modified without changing membrane voltage reveal that the ivabradine block depends on the current driving force, rather than voltage alone, a feature typical of block induced in inwardly rectifying K(+) channels by intracellular cations. Bound drug molecules do not detach from the binding site in the absence of inward current through channels, even if channels are open and the drug is therefore not "trapped" by closed gates. Our data suggest that permeation through f-channel pores occurs according to a multiion, single-file mechanism, and that block/unblock by ivabradine is coupled to ionic flow. The use-dependence resulting from specific features of I(f) block by ivabradine amplifies its rate reducing ability at high spontaneous rates and may be useful to clinical applications. PMID- 12084772 TI - Spontaneous transient outward currents arise from microdomains where BK channels are exposed to a mean Ca(2+) concentration on the order of 10 microM during a Ca(2+) spark. AB - Ca(2+) sparks are small, localized cytosolic Ca(2+) transients due to Ca(2+) release from sarcoplasmic reticulum through ryanodine receptors. In smooth muscle, Ca(2+) sparks activate large conductance Ca(2+)-activated K(+) channels (BK channels) in the spark microdomain, thus generating spontaneous transient outward currents (STOCs). The purpose of the present study is to determine experimentally the level of Ca(2+) to which the BK channels are exposed during a spark. Using tight seal, whole-cell recording, we have analyzed the voltage dependence of the STOC conductance (g((STOC))), and compared it to the voltage dependence of BK channel activation in excised patches in the presence of different [Ca(2+)]s. The Ca(2+) sparks did not change in amplitude over the range of potentials of interest. In contrast, the magnitude of g((STOC)) remained roughly constant from 20 to -40 mV and then declined steeply at more negative potentials. From this and the voltage dependence of BK channel activation, we conclude that the BK channels underlying STOCs are exposed to a mean [Ca(2+)] on the order of 10 microM during a Ca(2+) spark. The membrane area over which a concentration > or =10 microM is reached has an estimated radius of 150-300 nm, corresponding to an area which is a fraction of one square micron. Moreover, given the constraints imposed by the estimated channel density and the Ca(2+) current during a spark, the BK channels do not appear to be uniformly distributed over the membrane but instead are found at higher density at the spark site. PMID- 12084773 TI - Molecular proximity of Kv1.3 voltage-gated potassium channels and beta(1) integrins on the plasma membrane of melanoma cells: effects of cell adherence and channel blockers. AB - Tumor cell membranes have multiple components that participate in the process of metastasis. The present study investigates the physical association of beta1 integrins and Kv1.3 voltage-gated potassium channels in melanoma cell membranes using resonance energy transfer (RET) techniques. RET between donor-labeled anti beta1-integrin and acceptor-labeled anti-Kv1.3 channels was detected on LOX cells adherent to glass and fibronectin-coated coverslips. However, RET was not observed on LOX cells in suspension, indicating that molecular proximity of these membrane molecules is adherence-related. Several K(+) channel blockers, including tetraethylammonium, 4-aminopyridine, and verapamil, inhibited RET between beta1 integrins and Kv1.3 channels. However, the irrelevant K(+) channel blocker apamin had no effect on RET between beta1-integrins and Kv1.3 channels. Based on these findings, we speculate that the lateral association of Kv1.3 channels with beta1 integrins contributes to the regulation of integrin function and that channel blockers might affect tumor cell behavior by influencing the assembly of supramolecular structures containing integrins. PMID- 12084774 TI - Channel openings are necessary but not sufficient for use-dependent block of cardiac Na(+) channels by flecainide: evidence from the analysis of disease linked mutations. AB - Na(+) channel blockers such as flecainide have found renewed usefulness in the diagnosis and treatment of two clinical syndromes arising from inherited mutations in SCN5A, the gene encoding the alpha subunit of the cardiac voltage gated Na(+) channel. The Brugada syndrome (BrS) and the LQT-3 variant of the Long QT syndrome are caused by disease-linked SCN5A mutations that act to change functional and pharmacological properties of the channel. Here we have explored a set of SCN5A mutations linked both to BrS and LQT-3 to determine what disease modified channel properties underlie distinct responses to the Na(+) channel blocker flecainide. We focused on flecainide block that develops with repetitive channel activity, so-called use-dependent block (UDB). Our results indicate that mutation-induced changes in the voltage-dependence of channel availability (inactivation) may act as determinants of flecainide block. The data further indicate that UDB by flecainide requires channel opening, but is not likely due to open channel block. Rather, flecainide appears to interact with inactivation states that follow depolarization-induced channel opening, and mutation-induced changes in channel inactivation will alter flecainide block independent of the disease to which the mutation is linked. Analysis of flecainide block of mutant channels linked to these rare disorders has provided novel insight into the molecular determinants of drug action. PMID- 12084775 TI - Spermine block of the strong inward rectifier potassium channel Kir2.1: dual roles of surface charge screening and pore block. AB - Inward rectification in strong inward rectifiers such as Kir2.1 is attributed to voltage-dependent block by intracellular polyamines and Mg(2+). Block by the polyamine spermine has a complex voltage dependence with shallow and steep components and complex concentration dependence. To understand the mechanism, we measured macroscopic Kir2.1 currents in excised inside-out giant patches from Xenopus oocytes expressing Kir2.1, and single channel currents in the inside-out patches from COS7 cells transfected with Kir2.1. We found that as spermine concentration or voltage increased, the shallow voltage-dependent component of spermine block at more negative voltages was caused by progressive reduction in the single channel current amplitude, without a decrease in open probability. We attributed this effect to spermine screening negative surface charges involving E224 and E299 near the inner vestibule of the channel, thereby reducing K ion permeation rate. This idea was further supported by experiments in which increasing ionic strength also decreased Kir2.1 single channel amplitude, and by mutagenesis experiments showing that this component of spermine block decreased when E224 and E299, but not D172, were neutralized. The steep voltage-dependent component of block at more depolarized voltages was attributed to spermine migrating deeper into the pore and causing fast open channel block. A quantitative model incorporating both features showed excellent agreement with the steady-state and kinetic data. In addition, this model accounts for previously described substate behavior induced by a variety of Kir2.1 channel blockers. PMID- 12084776 TI - Hormonal regulation of the epithelial sodium channel ENaC: N or P(o)? PMID- 12084777 TI - cAMP increases density of ENaC subunits in the apical membrane of MDCK cells in direct proportion to amiloride-sensitive Na(+) transport. AB - Antidiuretic hormone and/or cAMP increase Na(+) transport in the rat renal collecting duct and similar epithelia, including Madin-Darby canine kidney (MDCK) cell monolayers grown in culture. This study was undertaken to determine if that increment in Na(+) transport could be explained quantitatively by an increased density of ENaC Na(+) channels in the apical membrane. MDCK cells with no endogenous ENaC expression were retrovirally transfected with rat alpha-, beta-, and gammaENaC subunits, each of which were labeled with the FLAG epitope in their extracellular loop as described previously (Firsov, D., L. Schild, I. Gautschi, A.-M. Merillat, E. Schneeberger, and B.C. Rossier. 1996. PROC: Natl. Acad. Sci. USA. 93:15370-15375). The density of ENaC subunits was quantified by specific binding of (125)I-labeled anti-FLAG antibody (M2) to the apical membrane, which was found to be a saturable function of M2 concentration with half-maximal binding at 4-8 nM. Transepithelial Na(+) transport was measured as the amiloride sensitive short-circuit current (AS-I(sc)) across MDCK cells grown on permeable supports. Specific M2 binding was positively correlated with AS-I(sc) measured in the same experiments. Stimulation with cAMP (20 microM 8-p-chlorothio-cAMP plus 200 microM IBMX) significantly increased AS-I(sc) from 11.2 +/- 1.3 to 18.1 +/- 1.3 microA/cm(2). M2 binding (at 1.7 nM M2) increased in direct proportion to AS I(sc) from 0.62 +/- 0.13 to 1.16 +/- 0.18 fmol/cm(2). Based on the concentration dependence of M2 binding, the quantity of Na(+) channels per unit of AS-I(sc) was calculated to be the same in the presence and absence of cAMP, 0.23 +/- 0.04 and 0.21 +/-0.05 fmol/microA, respectively. These values would be consistent with a single channel conductance of approximately 5 pS (typically reported for ENaC channels) only if the open probability is <0.02, i.e., less than one-tenth of the typical value. We interpret the proportional increases in binding and AS-I(sc) to indicate that the increased density of ENaC subunits in the apical membrane can account completely for the I(sc) increase produced by cAMP. PMID- 12084780 TI - Aging and the representation of spatial situations in narrative understanding. AB - Age differences in the construction of the situation model during text understanding were investigated. Situation model processing was measured in terms of the distance effect, the tendency for readers to process information about objects in a narrative more quickly when the objects are spatially closer to the protagonist than when they are farther away. To examine readers' ability to construct the model directly from the text, the distance effect was contrasted for objects that were either presented in a layout of the narrative setting prior to reading (learned objects) or introduced in the narrative itself (new objects). Both younger and older readers showed strong distance effects for learned objects. When objects had not been learned but were only introduced in the text, however, younger adults did not show a reliable distance effect. Older adults with high levels of comprehension did, lending support to the position that older readers differentially rely on the situation model for effective narrative understanding. PMID- 12084778 TI - Tail end of the s6 segment: role in permeation in shaker potassium channels. AB - The permeation pathway in voltage-gated potassium channels has narrow constrictions at both the extracellular and intracellular ends. These constrictions might limit the flux of cations from one side of the membrane to the other. The extracellular constriction is the selectivity filter, whereas the intracellular bundle crossing is proposed to act as the activation gate that opens in response to a depolarization. This four-helix bundle crossing is composed of S6 transmembrane segments, one contributed by each subunit. Here, we explore the cytoplasmic extension of the S6 transmembrane segment of Shaker potassium channels, just downstream from the bundle crossing. We substituted cysteine for each residue from N482 to T489 and determined the amplitudes of single channel currents and maximum open probability (P(o,max)) at depolarized voltages using nonstationary noise analysis. One mutant, F484C, significantly reduces P(o,max), whereas Y483C, F484C, and most notably Y485C, reduce single channel conductance (gamma). Mutations of residue Y485 have no effect on the Rb(+)/K(+) selectivity, suggesting a local effect on gamma rather than an allosteric effect on the selectivity filter. Y485 mutations also reduce pore block by tetrabutylammonium, apparently by increasing the energy barrier for blocker movement through the open activation gate. Replacing Rb(+) ions for K(+) ions reduces the amplitude of single channel currents and makes gamma insensitive to mutations of Y485. These results suggest that Rb(+) ions increase an extracellular energy barrier, presumably at the selectivity filter, thus making it rate limiting for flux of permeant ions. These results indicate that S6(T) residues have an influence on the conformation of the open activation gate, reflected in both the stability of the open state and the energy barriers it presents to ions. PMID- 12084781 TI - Working memory and online syntactic processing in Alzheimer's disease: studies with auditory moving window presentation. AB - Twenty patients with dementia of the Alzheimer's type (DAT) and 20 controls were tested on six tests of working memory and a test of online auditory sentence comprehension in which listening times for each phrase in the sentence, as well as the time required for an end-of-sentence plausibility judgment, were measured. The sentences differed in syntactic complexity. Patients had lower working memory scores than controls and performed more poorly on the plausibility judgments. However, patients were not more affected than controls by the syntactic complexity of a sentence in these judgments, and both groups showed similar effects of syntactic structure in the listening-time data. The increase in listening times at syntactically capacity-demanding points in complex sentences, compared with comparable points in matched simpler sentences, did not correlate with measures of working memory. The results indicate that early-stage DAT patients are not impaired in their ability to assign syntactic structure and to use it to determine aspects of sentence meaning, despite their reduced working memories. This provides evidence for a specialization within working memory for syntactic processing. PMID- 12084779 TI - Cysteine mutagenesis and computer modeling of the S6 region of an intermediate conductance IKCa channel. AB - Cysteine-scanning mutagenesis (SCAM) and computer-based modeling were used to investigate key structural features of the S6 transmembrane segment of the calcium-activated K(+) channel of intermediate conductance IKCa. Our SCAM results show that the interaction of [2-(trimethylammonium)ethyl] methanethiosulfonate bromide (MTSET) with cysteines engineered at positions 275, 278, and 282 leads to current inhibition. This effect was state dependent as MTSET appeared less effective at inhibiting IKCa in the closed (zero Ca(2+) conditions) than open state configuration. Our results also indicate that the last four residues in S6, from A283 to A286, are entirely exposed to water in open IKCa channels, whereas MTSET can still reach the 283C and 286C residues with IKCa maintained in a closed state configuration. Notably, the internal application of MTSET or sodium (2 sulfonatoethyl) methanethiosulfonate (MTSES) caused a strong Ca(2+)-dependent stimulation of the A283C, V285C, and A286C currents. However, in contrast to the wild-type IKCa, the MTSET-stimulated A283C and A286C currents appeared to be TEA insensitive, indicating that the MTSET binding at positions 283 and 286 impaired the access of TEA to the channel pore. Three-dimensional structural data were next generated through homology modeling using the KcsA structure as template. In accordance with the SCAM results, the three-dimensional models predict that the V275, T278, and V282 residues should be lining the channel pore. However, the pore dimensions derived for the A283-A286 region cannot account for the MTSET effect on the closed A283C and A286 mutants. Our results suggest that the S6 domain extending from V275 to V282 possesses features corresponding to the inner cavity region of KcsA, and that the COOH terminus end of S6, from A283 to A286, is more flexible than predicted on the basis of the closed KcsA crystallographic structure alone. According to this model, closure by the gate should occur at a point located between the T278 and V282 residues. PMID- 12084782 TI - Aging, cognitive complexity, and the fundamental attribution error. AB - Age differences in the prevalence and underpinnings of the fundamental attribution error were examined. Young, middle-aged, and older adults observed an actor providing responses to questions about political issues. In the no-choice condition, the actor was instructed how to respond, whereas in the choice condition, he was allowed to select his response. Consistent with previous research, middle-aged adults were less prone to the fundamental attribution error than were young and older adults. This was evidenced by their reduced tendency to make attitude attributions in the no-choice condition relative to the choice condition. Although high levels of both cognitive and attributional complexity were associated with reductions in attributional bias, complexity did not systematically account for the between-age-group differences in performance. It is suggested that the observed pattern of age effects is related to variations in both cognitive complexity and cognitive resources. PMID- 12084783 TI - Perceptual processing and search efficiency of young and older adults in a simple feature search task: a staircase approach. AB - The reasons that visual search may sometimes be difficult, especially for older adults, remain important research issues. This study investigated (a) whether age related differences can occur in simple-feature search, (b) if so, whether slowing adequately accounts for these differences, (c) whether other perceptual/cognitive factors are involved, and (d) the role of perceptual strategies. The authors tested 15 young adults (ages 18-30) and 15 older adults (ages 65-78). The target was a red disc presented among red diamonds in an array of 16 or 36 items. The forced-choice staircase procedure emphasized perceptual processing while deemphasizing decision-making and psychomotor processing. Although perceptual slowing may affect older adults' search performance, the perceptual slowing model is not simple, and other perceptual/cognitive factors, such as spatial resolution and distractibility, also are implicated. Moreover, perceptual strategies involving perceptual grouping or suppression of distractors play a key role in explaining why search efficiency is actually better for the larger set size. PMID- 12084784 TI - Temporal and reciprocal relationship between IADL/ADL disability and depressive symptoms in late life. AB - A strong association between functional disability and depressive symptoms in older people has frequently been reported. Some studies attribute this association to the disabling effects of depression, others to the depressogenic effects of physical health-related disability. The authors examined the reciprocal effects between depressive symptoms and functional disability and their temporal character in a community-based cohort of 753 older people with physical limitations who were assessed at yearly intervals. They compared structural equation models that differed in terms of direction and speed of effects between patient-reported disability in instrumental and basic activities of daily living (IADL/ADLs) and depressive symptoms. The association between disability and depression could be separated into three components: (a) a strong contemporaneous effect of change in disability on depressive symptoms, (b) a weaker 1-year lagged effect of change in depressive symptoms on disability (probably indirect through physical health), and (c) a weak correlation between the trait (or stable) components of depression and disability. IADL/ADL disability and depressive symptoms are thus mutually reinforcing over time. Compensatory forces like effective treatment and age-related adaptation may protect elders against this potential downward trend. To improve quality of life in elderly adults, treatment should target disability when it is new and depression when it is persistent. PMID- 12084785 TI - Evaluation of geriatric day care units: effects on patients and caregivers. AB - The aim of this study was to evaluate the effects of geriatric day care on patients and caregivers. Day care patients (n = 43) were compared with an untreated matched control group (n = 40). Matching criteria were age, gender, physical and mental health status, and socioeconomic status. The mean age of both groups was 79.5 years. Data were collected at three measurement points: T1 took place during the first 10 days of service use, T2 was conducted 6 months later, and T3 took place 9 months after service use began. In addition, a short follow up was conducted 6 months after T3. Well-being, dementia symptoms, health indicators, and activities of daily living were investigated. Individual growth curves of these patient variables were computed and analyzed with multivariate analyses of variance. The results show significant positive effects of day care on well-being and dementia symptoms. Patients in day care stabilized or improved on various measures, whereas the untreated control participants worsened. Follow up data showed a significant decline in health in the control group in comparison with the day care users. Results concerning effects on caregivers were not that clear. Individual change parameters in subjective well-being and burden did not differ between the two groups of caregivers in the longitudinal follow-up. In a semistructured interview, however, caregivers of patients in the treatment group reported substantial positive change due to use of day care. PMID- 12084786 TI - Fear of death in older adults: predictions from terror management theory. AB - Terror management theory asserts that death fear (fear of annihilation) is buffered by self-esteem and beliefs in literal and symbolic immortality achieved through participation in the cultural system. The aims of this study were to determine how variables suggested by the theory were related to fear of death measures. Participants were 123 Black and 265 White elders aged 60 to 100 years; they were assessed on the Multidimensional Fear of Death Scale (MFODS), self esteem, religiosity, locus of control, socioeconomic status, social support, and health. Regression analysis findings ( p <.05) offered partial support to the theory, with greater Fear of the Unknown (fear of annihilation) related to weaker religiosity, less social support, and greater externality; the effect of self esteem was mediated by externality. Other predictors were related to an overall fear score based on the remaining 7 MFODS subscales. Findings are interpreted in terms of changing sources of self-esteem in old age. PMID- 12084787 TI - Giving up and replacing activities in response to illness. AB - Data from a longitudinal study of 250 older adults were used to examine activity loss and replacement as a consequence of an important illness episode. Multiple regression analyses revealed that reductions in activity were predicted by physical factors, specifically illness chronicity and severity. In contrast, replacing lost activities was facilitated by social support and optimism and inhibited by a belief in the need to conserve physical resources. An examination of the long-term benefits of replacing activities revealed that older adults who replaced a lost activity had higher positive affect levels 1 year after illness onset than those who did not replace activities. Continuing activity during illness episodes can help maintain positive well-being over time. PMID- 12084788 TI - Cardiovascular risk factors and cognitive function in African Americans. AB - The present study examined the cross-sectional association of medically determined cardiovascular risk factors with cognitive function in 43 African Americans (aged 43-82 yr; 83% women). Measures of attention, memory, and executive functions were evaluated in relation to blood pressure (BP), total cholesterol, glycosylated hemoglobin (HbA1c), and fitness level (peak O(2)). Multiple regression analyses with age, education, number of antihypertensive medications, HbA1c, diastolic BP, and peak O(2) as predictors revealed significant (and marginally significant) associations between lower levels of fitness (peak O(2)) and poorer executive functions and delayed verbal memory. Antihypertensive medications were associated with poorer attention, but better delayed verbal memory. In addition, greater levels of HbA1c were positively related to attention. These results suggest that cardiovascular risk factors are important predictors of cognitive function among middle-aged and older African Americans. PMID- 12084789 TI - Housing quality and psychological well-being among the elderly population. AB - With exponential growth in the number of elderly Americans living independently in the community, it is important to better understand the implications of living conditions for psychological well-being among the elderly population. In a cross sectional study, housing quality is associated with positive affect among the older adults living independently in the community. Moreover, this relation is mediated by place attachment. Elderly residents of higher quality homes, independent of multiple sociodemographic factors (e.g., income, gender), feel more attached to their home, which, in turn, appears to account for the relation between housing quality and positive affect. PMID- 12084790 TI - Who expects to continue working after age 62? The retirement plans of couples. AB - OBJECTIVES: This study examines the individual, spousal, and household characteristics associated with the retirement expectations of husbands and wives. METHODS: Using data from the 1992 Health and Retirement Study, subjective probabilities of working full-time after reaching age 62 and age 65 are used to measure retirement expectations. The retirement expectations of husbands and wives are modeled simultaneously using a joint-generalized least-squares approach. RESULTS: Within a marriage, retirement expectations are shaped by individual, spousal, and household characteristics. We observe some gender differences in cross-spousal influence with wives' retirement expectations being more influenced by husbands' resources and constraints than vice versa. Nonetheless, individual and household factors associated with retirement expectations are widely shared by husbands and wives. DISCUSSION: Husbands and wives both respond to individual and joint constraints and opportunities when planning for retirement. Findings support that there is considerable overlap in retirement planning of husbands and wives during early parts of the retirement decision-making process. However, inequity in cross-spousal influences is a defining characteristic of retirement decision making. Implications for both policy makers and practitioners are briefly discussed. PMID- 12084791 TI - Transitions in living arrangements among elders in Japan: does health make a difference? AB - OBJECTIVE: This research evaluates the hypothesis that poor health triggers a change in living arrangements among elderly adults in Japan. METHODS: Data came from a national probability sample of 2,200 Japanese people aged 60 or older. Four surveys of this sample were conducted over a period of 9 years, from 1987 to 1996. Multinomial logit regression analyses were used to analyze the effects of demographic characteristics, socioeconomic status, and health on changes in living arrangements. RESULTS: Living arrangements among the Japanese elderly people remained quite stable over the 9-year period. Physical and mental health conditions were found to exert both direct and indirect effects on transitions in living arrangements. DISCUSSION: Poor health does trigger changes in living arrangement. Both physical (i.e., chronic conditions and functional status) and mental (i.e., depressed affect) health conditions play a role in such transitions. Because health conditions are correlated with competing risks of mortality, attrition, and proxy interview, health effects on changes in living arrangement are likely underestimated. PMID- 12084792 TI - Gender and ethnic/racial disparities in health care utilization among older adults. AB - OBJECTIVE: We examine the role of economic access in gender and ethnic/racial disparities in the use of health services among older adults. METHODS: Data from the 1993-1995 study on the Asset of Health Dynamics Among the Oldest Old (AHEAD) were used to investigate differences in the 2-year use of health services by gender and among non-Hispanic White versus minority (Hispanic and African American) ethnic/racial groups. Analyses account for predisposing factors, health needs, and economic access. RESULTS: African American men had fewer physician contacts; minority and non-Hispanic White women used fewer hospital or outpatient surgery services; minority men used less outpatient surgery; and Hispanic women were less likely to use nursing home care, compared with non-Hispanic White men, controlling for predisposing factors and measures of need. Although economic access was related to some medical utilization, it had little effect on gender/ethnic disparities for services covered by Medicare. However, economic access accounted for minority disparities in dental care, which is not covered by Medicare. DISCUSSION: Medicare plays a significant role in providing older women and minorities access to medical services. Significant gender and ethnic/racial disparities in use of medical services covered by Medicare were not accounted for by economic access among older adults with similar levels of health needs. Other cultural and attitudinal factors merit investigation to explain these gender/ethnic disparities. PMID- 12084793 TI - Patterns of family visiting with institutionalized elders: the case of dementia. AB - OBJECTIVES: The purpose of this study was to examine long-term trajectories of visits to nursing homes by family caregivers to persons suffering from dementia. METHODS: The data were obtained from a 5-year longitudinal study of family caregiving. This analysis used a subsample of caregivers who moved their relatives from home to a nursing home (n = 210). The frequency (times per week) and length (hours per week) of visits for up to 5 years after placement were examined in relation to select caregiver and care-recipient characteristics. A semiparametric, group-based method using a latent class mixture model was used to identify distinctive trajectories over time. RESULTS: The majority of caregivers maintained the frequency and length of their visits for extended periods of time. Five distinct patterns for frequency and three patterns for length were identified. Being a spouse, lower education, a close past relationship, a strong sentiment against placement, and living close to the facility predicted membership in groups visiting frequently and for longer times. Caregiver characteristics were more strongly associated with frequency, whereas care recipient characteristics were related to length. DISCUSSION: To a large extent, family visits to nursing home residents with dementia become established in the period immediately after relocation, which means that efforts to enhance continued family involvement should commence at the time of admission. PMID- 12084794 TI - The association of race and socioeconomic status with cardiovascular disease indicators among older adults in the health, aging, and body composition study. AB - OBJECTIVES: We hypothesized that older Black adults would have a higher prevalence of cardiovascular disease (CVD) than White adults, as indicated by elevated systolic blood pressure (SBP), low ankle-arm blood pressure index (AAI), and left ventricular hypertrophy (LVH). But, accounting for a broad interpretation of socioeconomic status (SES) (i.e., education, family income, home ownership, and other assets) would reduce these differences. METHODS: Data are from the Health, Aging, and Body Composition study, a longitudinal clinical research study of 3,075 well-functioning adults aged 70-79, in which 46% of women and 33% of men are Black. Logistic regression modeled racial and SES differences in CVD indicators. RESULTS: Being Black was significantly associated with elevated SBP (men only), low AAI, and LVH, and remained significant after accounting for each SES measure. The racial association with CVD was reduced the most by income for elevated SBP in men, other assets for low AAI in women and men, and other assets for LVH in men. DISCUSSION: Contrary to the age-as-leveler theory, being Black was strongly associated with CVD indicators, and accounting for SES did not reduce this association. Whether other SES measures, such as access to care, could explain the racial association remains to be explored. PMID- 12084795 TI - Cellular senescence and tissue aging in vivo. AB - A long-standing controversy concerns the relevance of cellular senescence, defined and observed as a cell culture phenomenon, to tissue aging in vivo. Here the evidence on this topic is reviewed. The main conclusions are as follows. First, telomere shortening, the principal known mediator of cellular senescence, occurs in many human tissues in aging. Second, it is not clear whether this results in cellular senescence or in some other cell fate (e.g., crisis). Third, rodents probably are not appropriate experimental models for these questions, because of important differences in telomere biology between rodent cells and cells from long-lived mammals (e.g., human or bovine cells). Fourth, better and more comprehensive observations on aging human tissues are needed to answer the question of the occurrence of senescent cells in tissues, and new experimental approaches are needed to elucidate the consequences of telomere shortening in tissues in aging. PMID- 12084796 TI - Replicative senescence revisited. AB - Forty years after its discovery, replicative senescence remains a rich source of information about cell-cycle regulation and the progression from a normal to a transformed phenotype. Effectors of this growth-arrested state are being discovered at a great pace. This review discusses the latest findings on the players responsible for establishing replicative senescence, as well as the associated telomere shortening. PMID- 12084798 TI - Mental stress response, arterial stiffness, and baroreflex sensitivity in healthy aging. AB - This study examined the relationship of pressor responses during mental stress to arterial stiffness and baroreflex sensitivity. Hemodynamic responses of 24 healthy individuals (51-86 years old) to two mental stress tasks (math and speech) were compared with common carotid artery mechanical stiffness and autonomic nervous system regulation of blood pressure as measured by using the modified Oxford technique. At the ages studied, no effect of age on stress task responsiveness, carotid stiffness, or baroreflex sensitivity was observed. Carotid stiffness and baroreflex sensitivity demonstrated a strong inverse relation. Change in heart rate during the speech task was correlated with arterial stiffness, and the increase in mean arterial pressure was associated with carotid stiffness and was inversely correlated to baroreflex sensitivity. These associations suggest that acute hemodynamic reactions to mental stress among healthy adults are determined, in part, by structural properties of arterial vessels and sensitivity of arterial baroreflex. These observations may provide a mechanistic link between the physiology of cardiovascular reactivity to stress and risk of cardiovascular events in middle-aged and older individuals. PMID- 12084797 TI - Age-associated cardiomyopathy in heterozygous carrier mice of a pathological mutation of carnitine transporter gene, OCTN2. AB - The purpose of this study was to test whether heterozygotes of juvenile visceral steatosis mice, a model for systemic carnitine deficiency, may develop age associated cardiomyopathy. Tissue morphological observations were carried out by light and electron microscopy to compare the heterozygous and age-matched control mice at periods of 1 and 2 years. Possible effects of the pathological mutation on lipid and glucose levels was also evaluated in humans and mice. Except mild increases in serum cholesterol levels in male heterozygous mice and humans, no changes were found in other factors, indicating that none of the confounding factors seems to be profound. Results demonstrated that heterozygous mice had larger left ventriclular myocyte diameters than the control mice. Morphological changes in cardiac muscles by electron microscopy revealed age-associated changes of lipid deposition and abnormal mitochondria in heterozygous mice. Two out of 60 heterozygous cohort and one out of nine heterozygous trim-kill mice had cardiac hypertrophy at ages older than 2 years. The present study and our previous work suggest that the carrier state of OCTN2 pathological mutations might be a risk factor for age-associated cardiomyopathy. PMID- 12084799 TI - Blood-sample processing for the study of age-dependent gene expression in peripheral blood mononuclear cells. AB - Although most new biogerontological studies seeking to identify longevity candidate genes and factors involved in successful human aging are population based, and likely to involve the collection of blood from extremely old individuals, to our knowledge no unified protocols have yet been published to describe a methodology permitting the simultaneous generation of different kinds of biological specimens derived from a single source of a very small volume of peripheral blood. Here we describe a method permitting the simultaneous generation of plasma, RNA, DNA, protein, fixed lymphocytes, and frozen blood aliquots from a single 10- to 30-ml blood sample obtained from donors of any age (10-102 years old), and we show that the quality and quantity of DNA, RNA, protein, and fixed lymphocytes obtained do not vary significantly with age. As is frequently observed, the older individuals have higher plasma proportions. PMID- 12084800 TI - Should hypercholesterolemia in older persons be treated to reduce cardiovascular events? PMID- 12084801 TI - The impact of the use of statins on the prevalence of dementia and the progression of cognitive impairment. AB - BACKGROUND: Previous evidence suggests that treatment with 3-hydroxy-3 methylglutaryl-coenzyme-A reductase inhibitors (statins) has a positive impact on dementia. We decided to investigate the association between the use of statins and the prevalence of dementia and statins' impact on the progression of cognitive impairment. METHODS: This is a case-control and a retrospective cohort study of a community-based ambulatory primary care geriatric practice. We included a convenience sample of all patients (N = 655, mean age 78.7 +/- 0.3 years, 85% Caucasian, 74% women) with hypercholesterolemia or dementia, or using statins. We compared those using statins with those who do not with respect to the clinical diagnosis of dementia and its subtypes and the progression of cognitive impairment. RESULTS: At the initial visit, 35% had dementia, and 17% were using statins. After covariate adjustments, patients on statins were less likely to have dementia (odds ratio [OR] for dementia based on composite definition = 0.23; 95% confidence interval [CI] [0.1-0.56], p =.001, OR Alzheimer's disease = 0.37; 95% CI [0.19-0.74], p =.005, OR vascular dementia = 0.25; 95% CI [0.08-0.85], p =.027). At follow-up, patients on statins showed an improvement on their Mini-Mental Status Examination score by 0.7 +/- 0.4 compared to a decline by 0.5 +/- 0.3 in controls, p =.025 (OR for no change or improvement on statins = 2.81; 95% CI [1.02-8.43], p =.045) and scored higher on the Clock Drawing Test (difference of 1.5 +/- 0.1, p =.036). CONCLUSIONS: The use of statins is associated with a lower prevalence of dementia and has a positive impact on the progression of cognitive impairment. PMID- 12084802 TI - Putting the "home" back in nursing home. PMID- 12084803 TI - The Eden Alternative: findings after 1 year of implementation. AB - BACKGROUND: This study examines the effects of the Eden Alternative (EA), a systematic introduction of pets, plants, and children into a nursing home, on the quality of life of nursing home residents. METHODS: Two nursing homes run by the same organization participated. The study site began implementing the EA in November 1998. The control site continued traditional care. Patient-level data from the Minimum Data Set (MDS), Version 2.0, and aggregate data based on staff reports were used to compare the residents at the two sites in terms of cognition, survival, immune function, functional status, and cost of care after 1 year. RESULTS: After adjusting for baseline differences, follow-up MDS data indicated that the Eden site had significantly greater proportions of residents who had fallen within the past 30 days (p =.011) and residents who were experiencing nutritional problems (p <.001). Staff report data indicated that, during the study period, the Eden site had significantly higher rates of residents requiring skilled nursing and hypnotic prescriptions, and more staff terminations and new hires. The control site had significantly higher rates of residents requiring anxiolytic prescriptions. CONCLUSIONS: The findings from this study indicate no beneficial effects of the EA in terms of cognition, functional status, survival, infection rate, or cost of care after 1 year. However, qualitative observations at the Eden site indicated that the change was positive for many staff as well as residents, suggesting that it may take longer than a year to demonstrate improvements attributable to the EA. PMID- 12084804 TI - The effects of animal-assisted therapy on loneliness in an elderly population in long-term care facilities. AB - BACKGROUND: Animal-assisted therapy (AAT) is claimed to have a variety of benefits, but almost all published results are anecdotal. We characterized the resident population in long-term care facilities desiring AAT and determined whether AAT can objectively improve loneliness. METHODS: Of 62 residents, 45 met inclusion criteria for the study. These 45 residents were administered the Demographic and Pet History Questionnaire (DPHQ) and Version 3 of the UCLA Loneliness Scale (UCLA-LS). They were then randomized into three groups (no AAT; AAT once/week; AAT three times/week; n = 15/group) and retested with the UCLA-LS near the end of the 6-week study. RESULTS: Use of the DPHQ showed residents volunteering for the study had a strong life-history of emotional intimacy with pets and wished that they currently had a pet. AAT was shown by analysis of covariance followed by pairwise comparison to have significantly reduced loneliness scores in comparison with the no AAT group. CONCLUSIONS: The desire for AAT strongly correlates with previous pet ownership. AAT reduces loneliness in residents of long-term care facilities. PMID- 12084805 TI - The autopsy in gerontological research: a retrospective. PMID- 12084806 TI - The future history of gerontological nursing. AB - Having established an academic and clinical infrastructure, geriatric nursing is well positioned to play a central role in improving the health of the nation's older adults now and in the future. Currently, whether working independently, in collaborative practice with physicians, or as members of geriatric teams, geriatric nurse practitioners and clinical specialists have been shown to improve care to older adults in the community, in hospitals, and in skilled nursing facilities. Sixty-three master's programs now prepare advanced practice geriatric nurses. Geriatric nurse researchers have contributed to our understanding of the most pressing problems that impact profoundly on the health and quality of life of older adults. Despite these advances, the number of geriatric nurse specialists remains small, with only 4200 certified specialists and a serious shortage of geriatric nursing faculty. Geriatric nursing is moving to ensure geriatric competency in all nurses who work with older adults in the future. The future should see the benefits of current efforts to infuse geriatric content into baccalaureate programs that prepare registered nurses, into master's programs that prepare adult and family nurse practitioners, and into the day-to day practice of the nation's 2.2 million practicing registered nurses. PMID- 12084807 TI - The effects of acute illness on ADL decline over 1 year in frail older adults with and without cognitive impairment. AB - BACKGROUND: Acute illness may lead to long-term losses in older adults' ability to independently perform activities of daily living (ADLs). The magnitude of these losses may differ for patients with and without cognitive impairment. These relationships have not been described for the frailest of older adults whose high rates of acute illness and cognitive impairment put them at the greatest risk for loss of ADL functioning. METHODS: We conducted a prospective study of 2593 patients enrolled in a nationwide medical and psychosocial program for frail, community-living, nursing home-eligible patients. We determined the independent and interactive effects of baseline cognitive impairment and admission for an acute illness on change in ADL functioning over 1 year. RESULTS: ADL decline over 1 year occurred in 53% of cognitively impaired patients who were admitted for an acute illness, 38% of cognitively impaired patients who were not admitted for an acute illness, 42% of noncognitively impaired patients who were admitted for an acute illness, and 25% of noncognitively impaired patients who were not admitted for an acute illness (p <.001). The amount of additional decline in ADLs associated with an admission for an acute illness was similar between patients with and without cognitive impairment (-.85 vs -.74; p for interaction =.86). Among patients who were admitted for an acute illness, significant decline in ADL functioning occurred only in the quarter surrounding the acute illness with no evidence of recovery in the months after the acute illness episode. CONCLUSIONS: Among frail older adults, loss of ADL functioning over 1 year is independently associated with both acute admission for an acute illness and cognitive impairment. Frail elders, especially those with cognitive impairment, are in need of interventions that reduce the long-term functional consequences of acute illness. PMID- 12084808 TI - Muscle size, strength, and bone geometry in the upper limbs of young and old men. AB - BACKGROUND: Bone loss in old men is associated with a decrease in muscle mass and strength. However, the influence of muscle size and strength on age-related changes in bone geometry has not been comprehensively described. Methods. Men in their third (group I, 23 +/- 3 y, n = 20), eighth (group II, 77 +/- 1 y, n = 10), and ninth (group III, 86 +/- 4 y, n = 13) decades of age were studied. The cross sectional area (CSA) of the elbow flexors, elbow extensors, and forearm muscles, the total area (TA), cortical area (CA), and medullary area (MA) of the midhumerus, and distal third of the radius and ulna (n = 7 group II; n = 6 group III) were measured with magnetic resonance imaging. The maximal isometric strength (MVC) of the elbow flexors and elbow extensors was also determined. RESULTS: The CSA and MVC of the arm muscles (elbow flexors plus elbow extensors) were less in group II (-17% and -22%) and III (-32% and -39%), respectively, compared to group I. However, forearm CSA was less (-21%) in group III only. The TA and MA of all bones were greater in the older groups. The CA of the humerus ( 14%) and ulna (-10%), but not the radius, was less in group III compared to group I, whereas CA was unchanged in group II. Stepwise multiple linear regression determined that arm muscle CSA (r = 0.52, p <.01) and forearm muscle CSA (r = 0.41, p <.05) provided the best prediction of CA in the humerus and forearm, respectively. CONCLUSIONS: Muscle size and strength are important determinants of CA in the humerus and forearm. The lower CA in the ninth decade may be explained, in part, by reduced bone strains due to a smaller muscle mass. PMID- 12084810 TI - Nonauditory determinants of self-perceived hearing problems among older adults: the role of stressful life conditions, neuroticism, and social resources. AB - BACKGROUND: The present study explored factors that influenced older individuals' subjective perception of hearing problems. In addition to objectively screened hearing ability, nonauditory factors such as stressful life conditions (visual impairment, chronic disease, disability, and recent stressful life events), neuroticism, and social resources were hypothesized to be predictors of self perceived hearing problems. METHODS: These hypotheses were tested with a hierarchical regression model using a stratified sample of 425 community-dwelling older individuals (mean age = 72.2). RESULTS: Individuals with more recent stressful life events, higher levels of neuroticism, and less emotional support reported greater hearing problems after controlling for objectively screened hearing. In addition to the main effects, a significant interaction was observed between neuroticism and screened hearing, indicating that the combination of poor hearing and high neuroticism increased the level of self-perceived hearing problems. CONCLUSIONS: The findings suggest that the effects of nonauditory factors should be taken into account in the application of self-assessed measures of hearing problems. PMID- 12084809 TI - Effects of transdermal testosterone on lipids and vascular reactivity in older men with low bioavailable testosterone levels. AB - BACKGROUND: Sex hormones are known to affect cholesterol levels and vascular tone in women. The effects of testosterone on cholesterol and vascular tone in men are less well understood. Low testosterone levels have been associated with higher cholesterol levels in epidemiologic studies, but testosterone replacement has resulted in variable changes in cholesterol levels. Similarly, clinical studies suggest that testosterone may be vasodilatory, but few studies have directly evaluated the effects of testosterone on vascular tone. METHODS: Sixty-seven men (mean age 76 +/- 4 years, range 65-87) with bioavailable testosterone levels below 4.44 nmol/l (lower limit for adult normal range) were randomized to receive transdermal testosterone (2-2.5 mg patches/d) or placebo patches for 1 year. Twenty-three men (34%) withdrew from the study; 44 men completed the trial. RESULTS: While total cholesterol, triglyceride, and low-density lipoprotein cholesterol levels did not significantly change during the year of therapy, high density lipoprotein (HDL) levels (p =.004) and, specifically, HDL(2) subfraction (p =.02) decreased in men receiving testosterone supplementation. Vascular tone was measured by brachial artery reactivity in 36 men. Endothelium-dependent brachial artery reactivity did not change from baseline measurements in men receiving transdermal testosterone (0.3 +/- 6.7% to 1.6 +/- 4.6%; p =.58) or in the placebo group (3.2 +/- 5.5% to 0.7 +/- 5.5%; p =.23). CONCLUSIONS: Transdermal testosterone decreased HDL(2) cholesterol but did not affect vascular reactivity in men older than 65 years selected for low testosterone levels. No study to date has addressed the direct relationship between testosterone replacement and cardiovascular events. PMID- 12084811 TI - The influence of intellectual disability on life expectancy. AB - BACKGROUND: To date, relatively few representative data have been available to health planners and advocacy groups on the life expectancy of people with intellectual disability. A study of trends in the survival profiles of people with intellectual disability was undertaken to assist in the planning of appropriate medical and support services. METHODS: Since 1953, the Disability Services Commission of Western Australia has maintained a database of persons diagnosed with intellectual disability. The database was used to calculate survival probabilities on a total of 8724 individuals, 7562 of whom were still alive at the time of sampling in December 2000. RESULTS: Kaplan-Meier survival plots showed a strong negative association between severity of intellectual disability and survival, with median life expectancies of 74.0, 67.6, and 58.6 years for people with mild, moderate, and severe levels of handicap. Significant negative associations also were observed with male gender, Indigenous Australian parentage, and individuals diagnosed with a specific genetic disorder. CONCLUSIONS: The findings indicate a major and expanding increase in the service requirements of this aging, intellectually disabled population during the past two generations. PMID- 12084812 TI - Falls in African American and white community-dwelling elderly residents. AB - BACKGROUND: Few studies have examined the relationship of race to falls. This study evaluated the association between potential risk factors and falls in a representative sample of 1049 African American and 1947 white participants of the second in-person wave of the Duke Established Populations for Epidemiologic Studies of the Elderly. METHODS: Information about sociodemographic characteristics, health-related behaviors, health status, visual function, and drug use was determined during baseline in-home interviews. Three years later, falls in the previous 12 months were assessed by self-report. RESULTS: One or more falls occurred in 22.2% of the participants. Nearly half the fallers reported more than one fall. Multivariable analysis revealed that African Americans were less likely than whites to have any fall (adjusted odds ratio [OR] 0.77, 95% confidence interval [CI] 0.62-0.94). Increased age and education, arthritis, diabetes, and history of broken bones were also significant (p <.05) independent risk factors for any fall. In multivariable analyses comparing those with two or more falls to those with none, again, increased age and education, arthritis, and diabetes were significant (p <.05) independent risk factors while smoking was protective. Race was not a significant predictor of multiple falls (adjusted OR 0.90, 95% CI 0.64-1.26). CONCLUSIONS: Similar sociodemographic characteristics and health problems appear to be important risk factors for any and multiple falls in community-dwelling African American and white elderly residents, with white elders at greater risk of one-time falls. PMID- 12084813 TI - SKIP is an indispensable factor for Caenorhabditis elegans development. AB - SKI-binding protein (SKIP) is a transcription cofactor present in all eukaryotes. Here we show that SKIP is a unique protein that is required for Caenorhabditis elegans viability and development. Expression of CeSKIP (skp-1) assayed by RT-PCR and by GFP fluorescence in transgenic lines starts in embryos and continues to adulthood. Loss of CeSKIP activity by RNA-mediated inhibition results in early embryonic arrest similar to that seen following inhibition of RNA polymerase II. RNA polymerase II phosphorylation appears normal early in CeSKIP RNA-mediated inhibition treated embryos although the expression of several embryonic GFP reporter genes is severely restricted or absent. Our data suggest that CeSKIP is an essential component of many RNA polymerase II transcription complexes and is indispensable for C. elegans development. PMID- 12084814 TI - Adeno-associated viral vector-mediated hypoxia response element-regulated gene expression in mouse ischemic heart model. AB - Intramyocardial injection of genes encoding angiogenic factors could provide a useful approach for the treatment of ischemic heart disease. However, uncontrolled expression of angiogenic factors in vivo may cause some unwanted side effects, such as hemangioma formation, retinopathy, and arthritis. It may also induce occult tumor growth and artherosclerotic plaque progression. Because hypoxia-inducible factor 1 is up-regulated in a variety of hypoxic conditions and it regulates gene expression by binding to a cis-acting hypoxia-responsive element (HRE), we propose to use HRE, found in the 3' end of the erythropoietin gene to control gene expression in ischemic myocardium. A concatemer of nine copies of the consensus sequence of HRE isolated from the erythropoietin enhancer was used to mediate hypoxia induction. We constructed two adeno-associated viral vectors in which LacZ and vascular endothelial growth factor (VEGF) expressions were controlled by this HRE concatemer and a minimal simian virus 40 promoter. Both LacZ and VEGF expression were induced by hypoxia and/or anoxia in several cell lines transduced with these vectors. The functions of these vectors in ischemic myocardium were tested by injecting them into normal and ischemic mouse myocardium created by occlusion of the left anterior descending coronary artery. The expression of LacZ gene was induced eight times and of VEGF 20 times in ischemic myocardium compared with normal myocardium after the viral vector transduction. Hence, HRE is a good candidate for the control of angiogenic factor gene expression in ischemic myocardium. PMID- 12084816 TI - The genome-wide expression response to telomerase deletion in Saccharomyces cerevisiae. AB - Loss of the protective function of telomeres has previously been hypothesized to cause a DNA damage response. Here, we report a genome-wide expression response, the telomerase deletion response (TDR), that occurs when telomeres can no longer be maintained by telomerase. The TDR shares features with other DNA damage responses and the environmental stress response. Unexpectedly, another feature of the TDR is the up-regulation of energy production genes, accompanied by a proliferation of mitochondria. Finally, a discrete set of genes, the "telomerase deletion signature", is uniquely up-regulated in the TDR but not under other conditions of stress and DNA damage that have been reported. The telomerase deletion signature genes define new candidates for involvement in cellular responses to altered telomere structure or function. PMID- 12084815 TI - Interactions between Eph kinases and ephrins provide a mechanism to support platelet aggregation once cell-to-cell contact has occurred. AB - Eph kinases are receptor tyrosine kinases whose ligands, the ephrins, are also expressed on the surface of cells. Interactions between Eph kinases and ephrins on adjacent cells play a central role in neuronal patterning and vasculogenesis. Here we examine the expression of ephrins and Eph kinases on human blood platelets and explore their role in the formation of the hemostatic plug. The results show that human platelets express EphA4 and EphB1, and the ligand, ephrinB1. Forced clustering of EphA4 or ephrinB1 led to cytoskeletal reorganization, adhesion to fibrinogen, and alpha-granule secretion. Clustering of ephrinB1 also caused activation of the Ras family member, Rap1B. In platelets that had been activated by ADP and allowed to aggregate, EphA4 formed complexes with two tyrosine kinases, Fyn and Lyn, and the cell adhesion molecule, L1. Blockade of Eph/ephrin interactions prevented the formation of these complexes and caused platelet aggregation at low ADP concentrations to become more readily reversible. We propose that when sustained contacts between platelets have occurred in response to agonists such as collagen, ADP, and thrombin, the binding of ephrins to Eph kinases on adjacent platelets provides a mechanism to perpetuate signaling and promote stable platelet aggregation. PMID- 12084817 TI - A protein kinase B-dependent and rapamycin-sensitive pathway controls skeletal muscle growth but not fiber type specification. AB - Nerve activity controls fiber size and fiber type in skeletal muscle, but the underlying molecular mechanisms remain largely unknown. We have previously shown that Ras-mitogen-activated protein kinase and calcineurin control fiber type but not fiber size in regenerating rat skeletal muscle. Here we report that constitutively active protein kinase B (PKB), also known as Akt, increases fiber size and prevents denervation atrophy in regenerating and adult rat muscles but does not affect fiber type profile. The coexistence of hypertrophic muscle fibers overexpressing activated PKB with normal-size untransfected fibers within the same muscle points to a cell-autonomous control of muscle growth by PKB. The physiological role of this pathway is confirmed by the finding that PKB kinase activity and phosphorylation status are significantly increased in innervated compared with denervated regenerating muscles in parallel with muscle growth. Muscle fiber hypertrophy induced by activated PKB and by a Ras double mutant (RasV12C40) that activates selectively the phosphoinositide 3-kinase-PKB pathway is completely blocked by rapamycin, showing that the mammalian target of rapamycin kinase is the major downstream effector of this pathway in the control of muscle fiber size. On the other hand, nerve activity-dependent growth of regenerating muscle is only partially inhibited by dominant negative PKB and rapamycin, suggesting that other nerve-dependent signaling pathways are involved in muscle growth. The present results support the notion that fiber size and fiber type are regulated by nerve activity through different mechanisms. PMID- 12084818 TI - Direct detection of potential selenium delivery proteins by using an Escherichia coli strain unable to incorporate selenium from selenite into proteins. AB - Selenium can be metabolized for protein synthesis by two major pathways in vivo. In a specific pathway it can be inserted into polypeptide chains as the amino acid selenocysteine, as directed by the UGA codon. Alternatively, selenium can be substituted for sulfur to generate the free amino acids selenocysteine and selenomethionine, and these are incorporated nonspecifically into proteins in place of cysteine and methionine, respectively. A mutant strain of Escherichia coli was constructed that is deficient in utilization of inorganic selenium for both specific and nonspecific pathways of selenoprotein synthesis. Disruption of the cysK gene prevented synthesis of free cysteine and selenocysteine from inorganic S and Se precursors. Inactivation of the selD gene prevented synthesis of selenophosphate, the reactive selenium donor, required for the specific incorporation pathway. As expected, the double mutant strain, RL165 Delta selD, when grown anaerobically in LB + glucose medium containing (75)SeO(3)(2-), failed to synthesize selenium-dependent formate dehydrogenase H and seleno-tRNAs. However, it incorporated 24% as much selenium as the wild-type strain. Selenium in the deficient strain was bound to five different proteins. A 39-kDa species was identified as glyceraldehyde-3-phosphate dehydrogenase. It is possible that selenium was bound as a perselenide derivative to the reactive cysteine residue of this enzyme. A 28-kDa protein identified as deoxyribose phosphate aldolase also contained bound selenium. These (75)Se-labeled proteins may have alternate roles as selenium delivery proteins. PMID- 12084819 TI - Live-cell imaging reveals divergent intracellular dynamics of polyglutamine disease proteins and supports a sequestration model of pathogenesis. AB - Protein misfolding and aggregation are central features of the polyglutamine neurodegenerative disorders, but the dynamic properties of expanded polyglutamine proteins are poorly understood. Here, we use fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) with green fluorescent protein fusion proteins to study polyglutamine protein kinetics in living cells. Our results reveal markedly divergent mobility states for an expanded polyglutamine protein, ataxin-3, and establish that nuclear inclusions formed by this protein are aggregates. Additional studies of green fluorescent protein-tagged cAMP response element binding protein coexpressed with either of two mutant polyglutamine proteins, ataxin-3 and huntingtin, support a model of disease in which coaggregation of transcriptional components contributes to pathogenesis. Finally, studies of a third polyglutamine disease protein, ataxin 1, reveal unexpected heterogeneity in the dynamics of inclusions formed by different disease proteins, a finding which may help explain disease-specific elements of pathogenesis in these neurodegenerative disorders. PMID- 12084820 TI - Probing the mysteries of lignin biosynthesis: the crystal structure of caffeic acid/5-hydroxyferulic acid 3/5-O-methyltransferase provides new insights. PMID- 12084821 TI - Contrapuntal networks of gene expression during Arabidopsis seed filling. AB - We have used cDNA microarrays to examine changes in gene expression during Arabidopsis seed development and to compare wild-type and mutant wrinkled1 (wri1) seeds that have an 80% reduction in oil. Between 5 and 13 days after flowering, a period preceding and including the major accumulation of storage oils and proteins, approximately 35% of the genes represented on the array changed at least twofold, but a larger fraction (65%) showed little or no change in expression. Genes whose expression changed most tended to be expressed more in seeds than in other tissues. Genes related to the biosynthesis of storage components showed several distinct temporal expression patterns. For example, a number of genes encoding core fatty acid synthesis enzymes displayed a bell shaped pattern of expression between 5 and 13 days after flowering. By contrast, the expression of storage proteins, oleosins, and other known abscisic acid regulated genes increased later and remained high. Genes for photosynthetic proteins followed a pattern very similar to that of fatty acid synthesis proteins, implicating a role in CO(2) refixation and the supply of cofactors for oil synthesis. Expression profiles of key carbon transporters and glycolytic enzymes reflected shifts in flux from cytosolic to plastid metabolism. Despite major changes in metabolism between wri1 and wild-type seeds, <1% of genes differed by more than twofold, and most of these were involved in central lipid and carbohydrate metabolism. Thus, these data define in part the downstream responses to disruption of the WRI1 gene. PMID- 12084822 TI - Functional analysis of a DNA-shuffled movement protein reveals that microtubules are dispensable for the cell-to-cell movement of tobacco mosaic virus. AB - Microtubules interact strongly with the viral movement protein (MP) of Tobacco mosaic virus (TMV) and are thought to transport the viral genome between plant cells. We describe a functionally enhanced DNA-shuffled movement protein (MP(R3)) that remained bound to the vertices of the cortical endoplasmic reticulum, showing limited affinity for microtubules. A single amino acid change was shown to confer the MP(R3) phenotype. Disruption of the microtubule cytoskeleton in situ with pharmacological agents, or by silencing of the alpha-tubulin gene, had no significant effect on the spread of TMV vectors expressing wild-type MP (MP(WT)) and did not prevent the accumulation of MP(WT) in plasmodesmata. Thus, cell-to-cell trafficking of TMV can occur independently of microtubules. The MP(R3) phenotype was reproduced when infection sites expressing MP(WT) were treated with a specific proteasome inhibitor, indicating that the degradation of MP(R3) is impaired. We suggest that the improved viral transport functions of MP(R3) arise from evasion of a host degradation pathway. PMID- 12084823 TI - IRT1, an Arabidopsis transporter essential for iron uptake from the soil and for plant growth. AB - Plants are the principal source of iron in most diets, yet iron availability often limits plant growth. In response to iron deficiency, Arabidopsis roots induce the expression of the divalent cation transporter IRT1. Here, we present genetic evidence that IRT1 is essential for the uptake of iron from the soil. An Arabidopsis knockout mutant in IRT1 is chlorotic and has a severe growth defect in soil, leading to death. This defect is rescued by the exogenous application of iron. The mutant plants do not take up iron and fail to accumulate other divalent cations in low-iron conditions. IRT1-green fluorescent protein fusion, transiently expressed in culture cells, localized to the plasma membrane. We also show, through promoter::beta-glucuronidase analysis and in situ hybridization, that IRT1 is expressed in the external cell layers of the root, specifically in response to iron starvation. These results clearly demonstrate that IRT1 is the major transporter responsible for high-affinity metal uptake under iron deficiency. PMID- 12084825 TI - Mutations in the Dof zinc finger genes DAG2 and DAG1 influence with opposite effects the germination of Arabidopsis seeds. AB - We describe the Arabidopsis gene DAG2 encoding a Dof zinc finger protein and show that it is involved in the control of seed germination. An Arabidopsis mutant line with a T-DNA insertion in DAG2 isolated by reverse genetics produces seeds that are substantially more dependent than the wild type on the physical stimuli light and cold treatment-that promote germination. Mutant dag2 seeds also are less sensitive to the germination-promotive effect of gibberellins, because a 10 fold higher amount of gibberellins is needed to restore germination when endogenous gibberellin biosynthesis is blocked. The seed germination characteristics of the dag2 mutant are opposite to those of dag1, a knockout mutant of another Dof gene (DAG1) that we showed previously to be involved in the control of seed germination, and are similar to those of plants that overexpress DAG1. The promoter of the DAG2 gene is active specifically in the vascular system of the mother plant but not in the embryo, and segregation analysis indicates that the effect of the dag2 mutation is maternal. Both characteristics are in common with DAG1; additionally, the DAG1 and DAG2 proteins share high sequence homology and an identical zinc finger domain. These data suggest, and the germination phenotype of the double mutant is compatible with, a model whereby the zinc finger proteins DAG1 and DAG2 act on a maternal switch that controls seed germination, possibly by regulating the same gene(s). PMID- 12084824 TI - A mitochondrial complex I defect impairs cold-regulated nuclear gene expression. AB - To study low-temperature signaling in plants, we previously screened for cold stress response mutants using bioluminescent Arabidopsis plants that express the firefly luciferase reporter gene driven by the stress-responsive RD29A promoter. Here, we report on the characterization and cloning of one mutant, frostbite1 (fro1), which shows reduced luminescence induction by cold. fro1 plants display reduced cold induction of stress-responsive genes such as RD29A, KIN1, COR15A, and COR47. fro1 leaves have a reduced capacity for cold acclimation, appear water soaked, leak electrolytes, and accumulate reactive oxygen species constitutively. FRO1 was isolated through positional cloning and found to encode a protein with high similarity to the 18-kD Fe-S subunit of complex I (NADH dehydrogenase, EC 1.6.5.3) in the mitochondrial electron transfer chain. Confocal imaging shows that the FRO1:green fluorescent protein fusion protein is localized in mitochondria. These results suggest that cold induction of nuclear gene expression is modulated by mitochondrial function. PMID- 12084826 TI - Structural basis for the modulation of lignin monomer methylation by caffeic acid/5-hydroxyferulic acid 3/5-O-methyltransferase. AB - Caffeic acid/5-hydroxyferulic acid 3/5-O-methyltransferase (COMT) from alfalfa is an S-adenosyl-L-Met-dependent O-methyltransferase involved in lignin biosynthesis. COMT methylates caffeoyl- and 5-hydroxyferuloyl-containing acids, aldehydes, and alcohols in vitro while displaying a kinetic preference for the alcohols and aldehydes over the free acids. The 2.2-A crystal structure of COMT in complex with S-adenosyl-L-homocysteine (SAH) and ferulic acid (ferulate form), as well as the 2.4-A crystal structure of COMT in complex with SAH and 5 hydroxyconiferaldehyde, provide a structural understanding of the observed substrate preferences. These crystal structures identify residues lining the active site surface that contact the substrates. Structurally guided site directed mutagenesis of active site residues was performed with the goal of altering the kinetic preferences for physiological substrates. The kinetic parameters of the COMT mutants versus wild-type enzyme are presented, and coupled with the high-resolution crystal structures, they will serve as a starting point for the in vivo manipulation of lignin monomers in transgenic plants. Ultimately, this structurally based approach to metabolic engineering will allow the further alteration of the lignin biosynthetic pathway in agronomically important plants. This approach will lead to a better understanding of the in vivo operation of the potential metabolic grid for monolignol biosynthesis. PMID- 12084827 TI - Dark-stimulated calcium ion fluxes in the chloroplast stroma and cytosol. AB - Using transgenic Nicotiana plumbaginifolia seedlings in which the calcium reporter aequorin is targeted to the chloroplast stroma, we found that darkness stimulates a considerable flux of Ca(2+) into the stroma. This Ca(2+) flux did not occur immediately after the light-to-dark transition but began approximately 5 min after lights off and increased to a peak at approximately 20 to 30 min after the onset of darkness. Imaging of aequorin emission confirmed that the dark stimulated luminescence emanated from chloroplast-containing tissues of the seedling. The magnitude of the Ca(2+) flux was proportional to the duration of light exposure (24 to 120 h) before lights off; the longer the duration of light exposure, the larger the dark-stimulated Ca(2+) flux. On the other hand, the magnitude of the dark-stimulated Ca(2+) flux did not appear to vary as a function of circadian time. When seedlings were maintained on a 24-h light/dark cycle, there was a stromal Ca(2+) burst after lights off every day. Moreover, the waveform of the Ca(2+) spike was different during long-day versus short-day light/dark cycles. The dark-stimulated Ca(2+) flux into the chloroplastidic stroma appeared to affect transient changes in cytosolic Ca(2+) levels. DCMU, an inhibitor of photosynthetic electron transport, caused a significant increase in stromal Ca(2+) levels in the light but did not affect the magnitude of the dark stimulated Ca(2+) flux. This robust Ca(2+) flux likely plays regulatory roles in the sensing of both light/dark transitions and photoperiod. PMID- 12084828 TI - Membrane protein transport between the endoplasmic reticulum and the Golgi in tobacco leaves is energy dependent but cytoskeleton independent: evidence from selective photobleaching. AB - The mechanisms that control protein transport between the endoplasmic reticulum (ER) and the Golgi apparatus are poorly characterized in plants. Here, we examine in tobacco leaves the structural relationship between Golgi and ER membranes using electron microscopy and demonstrate that Golgi membranes contain elements that are in close association and/or in direct contact with the ER. We further visualized protein trafficking between the ER and the Golgi using Golgi marker proteins tagged with green fluorescent protein. Using photobleaching techniques, we showed that Golgi membrane markers constitutively cycle to and from the Golgi in an energy-dependent and N-ethylmaleimide-sensitive manner. We found that membrane protein transport toward the Golgi occurs independently of the cytoskeleton and does not require the Golgi to be motile along the surface of the ER. Brefeldin A treatment blocked forward trafficking of Golgi proteins before their redistribution into the ER. Our results indicate that in plant cells, the Golgi apparatus is a dynamic membrane system whose components continuously traffic via membrane trafficking pathways regulated by brefeldin A- and N ethylmaleimide-sensitive machinery. PMID- 12084829 TI - Developing seeds of Arabidopsis store different minerals in two types of vacuoles and in the endoplasmic reticulum. AB - Mineral-accumulating compartments in developing seeds of Arabidopsis were studied using high-pressure-frozen/freeze-substituted samples. Developing seeds store minerals in three locations: in the protein storage vacuoles of the embryo, and transiently in the endoplasmic reticulum (ER) and vacuolar compartments of the chalazal endosperm. Energy dispersive x-ray spectroscopy and enzyme treatments suggest that the minerals are stored as phytic acid (myo-inositol-1,2,3,4,5,6 hexakisphosphate) salts in all three compartments, although they differ in cation composition. Whereas embryo globoids contain Mg, K, and Ca as cations, the chalazal ER deposits show high levels of Mn, and the chalazal vacuolar deposits show high levels of Zn. The appearance of the first Zn-phytate crystals coincides with the formation of network-like extensions of the chalazal vacuoles. The core of these networks consists of a branched network of tubular ER membranes, which are separated from the delineating tonoplast membranes by a layer of cytosolic material. Degradation of the networks starts with the loss of the cytosol and is followed by the retraction of the ER, generating a network of collapsed tonoplast membranes that are resorbed. Studies of fertilized fis2 seeds, which hyperaccumulate Zn-phytate crystals in the chalazal vacuolar compartments, suggest that only the intact network is active in mineral sequestration. Mineral determination analysis and structural observations showed that Zn and Mn are mobilized from the endosperm to the embryo at different developmental stages. Thus, Zn appears to be removed from the endosperm at the late globular stage, and Mn stores appear to be removed at the late bent-cotyledon stage of embryo development. The disappearance of the Mn-phytate from the endosperm coincides with the accumulation of two major Mn binding proteins in the embryo, the 33-kD protein from the oxygen-evolving complex of photosystem II and the Mn superoxide dismutase. The possible functions of transient heavy metal storage in the chalazal endosperm are discussed. A model showing how phytic acid, a potentially cytotoxic molecule, is transported from its site of synthesis, the ER, to the different mineral storage sites is presented. PMID- 12084830 TI - Molecular cloning and characterization of glucanase inhibitor proteins: coevolution of a counterdefense mechanism by plant pathogens. AB - A characteristic plant response to microbial attack is the production of endo beta-1,3-glucanases, which are thought to play an important role in plant defense, either directly, through the degradation of beta-1,3/1,6-glucans in the pathogen cell wall, or indirectly, by releasing oligosaccharide elicitors that induce additional plant defenses. We report the sequencing and characterization of a class of proteins, termed glucanase inhibitor proteins (GIPs), that are secreted by the oomycete Phytophthora sojae, a pathogen of soybean, and that specifically inhibit the endoglucanase activity of their plant host. GIPs are homologous with the trypsin class of Ser proteases but are proteolytically nonfunctional because one or more residues of the essential catalytic triad is absent. However, specific structural features are conserved that are characteristic of protein-protein interactions, suggesting a mechanism of action that has not been described previously in plant pathogen studies. We also report the identification of two soybean endoglucanases: EGaseA, which acts as a high affinity ligand for GIP1; and EGaseB, with which GIP1 does not show any association. In vitro, GIP1 inhibits the EGaseA-mediated release of elicitor active glucan oligosaccharides from P. sojae cell walls. Furthermore, GIPs and soybean endoglucanases interact in vivo during pathogenesis in soybean roots. GIPs represent a novel counterdefensive weapon used by plant pathogens to suppress a plant defense response and potentially function as important pathogenicity determinants. PMID- 12084831 TI - Expression of the IRT1 metal transporter is controlled by metals at the levels of transcript and protein accumulation. AB - Iron, an essential nutrient, is not readily available to plants because of its low solubility. In addition, iron is toxic in excess, catalyzing the formation of hydroxyl radicals that can damage cellular constituents. Consequently, plants must carefully regulate iron uptake so that iron homeostasis is maintained. The Arabidopsis IRT1 gene is the major transporter responsible for high-affinity iron uptake from the soil. Here, we show that the steady state level of IRT1 mRNA was induced within 24 h after transfer of plants to iron-deficient conditions, with protein levels peaking 72 h after transfer. IRT1 mRNA and protein were undetectable 12 h after plants were shifted back to iron-sufficient conditions. Overexpression of IRT1 did not confer dominant gain-of-function enhancement of metal uptake. Analysis of 35S-IRT1 transgenic plants revealed that although IRT1 mRNA was expressed constitutively in these plants, IRT1 protein was present only in the roots when iron is limiting. Under these conditions, plants that overexpressed IRT1 accumulated higher levels of cadmium and zinc than wild-type plants, indicating that IRT1 is responsible for the uptake of these metals and that IRT1 protein levels are indeed increased in these plants. Our results suggest that the expression of IRT1 is controlled by two distinct mechanisms that provide an effective means of regulating metal transport in response to changing environmental conditions. PMID- 12084832 TI - TRANSPARENT TESTA GLABRA2, a trichome and seed coat development gene of Arabidopsis, encodes a WRKY transcription factor. AB - Mutants of a new gene, TRANSPARENT TESTA GLABRA2 (TTG2), show disruptions to trichome development and to tannin and mucilage production in the seed coat. The gene was tagged by the endogenous transposon Tag1 and shown to encode a WRKY transcription factor. It is the first member of this large, plant-specific family known to control morphogenesis. The functions of all other WRKY genes revealed to date involve responses to pathogen attack, mechanical stress, and senescence. TTG2 is strongly expressed in trichomes throughout their development, in the endothelium of developing seeds (in which tannin is later generated) and subsequently in other layers of the seed coat, and in the atrichoblasts of developing roots. TTG2 acts downstream of the trichome initiation genes TTG1 and GLABROUS1, although trichome expression of TTG2 continues to occur if they are inactivated. Later, TTG2 shares functions with GLABRA2 in controlling trichome outgrowth. In the seed coat, TTG2 expression requires TTG1 function in the production of tannin. Finally, TTG2 also may be involved in specifying atrichoblasts in roots redundantly with other gene(s) but independently of TTG1 and GLABRA2. PMID- 12084833 TI - In vivo interaction between NPR1 and transcription factor TGA2 leads to salicylic acid-mediated gene activation in Arabidopsis. AB - The Arabidopsis NPR1 protein is a key regulator of salicylic acid (SA)-mediated gene expression in systemic acquired resistance. Based on yeast two-hybrid analysis, NPR1 has been suggested to interact with members of the TGA family of transcription factors, including TGA2 (AHBP-1b). However, genetic evidence demonstrating that the NPR1-TGA interaction occurs in planta is still lacking, and the role of this interaction in SA-mediated gene activation has yet to be determined. In this study, we expressed a truncated form of TGA2 in Arabidopsis and found that the resulting transgenic lines displayed phenotypes similar to those of npr1 mutants. This dominant-negative effect of the TGA2 mutant shows that TGA2 and NPR1 interact in planta. We also present biochemical evidence indicating that this interaction is specific and enhanced by SA treatment. Moreover, using a chimera reporter system, we found that a chimeric TGA2GAL4 transcription factor activated a UAS(GAL)::GUS reporter gene in response to SA and that this activation was abolished in the npr1 mutant. NPR1 is required for the DNA binding activity of the transcription factor. These genetic data clearly demonstrate that TGA2 is a SA-responsive and NPR1-dependent transcription activator. PMID- 12084834 TI - The homologous ABI5 and EEL transcription factors function antagonistically to fine-tune gene expression during late embryogenesis. AB - In Arabidopsis, the basic leucine zipper transcription factor ABI5 activates several late embryogenesis-abundant genes, including AtEm1 and AtEm6. However, the expression of many other seed maturation genes is independent of ABI5. We investigated the possibility that ABI5 homologs also participate in the regulation of gene expression during seed maturation. We identified 13 ABI5 related genes in the Arabidopsis genomic sequence. RNA gel blot analysis showed that seven of these genes are active during seed maturation and that they display distinct expression kinetics. We isolated and characterized two mutant alleles of one of these genes, AtbZIP12/EEL. Unlike abi5, the eel mutations did not inhibit the expression of any of the maturation marker genes that we monitored. On the contrary, the accumulation of the AtEm1 and AtEm6 mRNAs was enhanced in eel mutant seeds compared with wild-type seeds. Gel mobility shift assays, combined with analysis of the genetic interactions among the eel and abi5 mutations, indicated that ABI5 and EEL compete for the same binding sites within the AtEm1 promoter. This study illustrates how two homologous transcription factors can play antagonistic roles to fine-tune gene expression. PMID- 12084835 TI - Jasmonate response locus JAR1 and several related Arabidopsis genes encode enzymes of the firefly luciferase superfamily that show activity on jasmonic, salicylic, and indole-3-acetic acids in an assay for adenylation. AB - Jasmonic acid (JA) and related cyclopentanones are critical plant signaling molecules, but their mode of action at the molecular level is unclear. A map based approach was used to identify the defective gene in the Arabidopsis JA response mutant jar1-1. JAR1 is 1 of 19 closely related Arabidopsis genes that are similar to the auxin-induced soybean GH3 gene. Analysis of fold predictions for this protein family suggested that JAR1 might belong to the acyl adenylate forming firefly luciferase superfamily. These enzymes activate the carboxyl groups of a variety of substrates for their subsequent biochemical modification. An ATP-PPi isotope exchange assay was used to demonstrate adenylation activity in a glutathione S-transferase-JAR1 fusion protein. Activity was specific for JA, suggesting that covalent modification of JA is important for its function. Six other Arabidopsis genes were specifically active on indole-3-acetic acid (IAA), and one was active on both IAA and salicylic acid. These findings suggest that the JAR1 gene family is involved in multiple important plant signaling pathways. PMID- 12084836 TI - The plastidic 2-cysteine peroxiredoxin is a target for a thioredoxin involved in the protection of the photosynthetic apparatus against oxidative damage. AB - The chloroplastic drought-induced stress protein of 32 kD (CDSP32) is composed of two thioredoxin modules and is induced by environmental and oxidative stress conditions. We investigated whether the plastidic protein BAS1, which is related to eubacterial 2-Cys peroxiredoxin, is a target for CDSP32. Using a CDSP32 active site mutant, we showed that the BAS1 and CDSP32 proteins form a mixed disulfide complex in vitro. Moreover, affinity chromatography indicated that BAS1 is a major target for CDSP32 in chloroplasts. CDSP32 was able to reduce BAS1 in vitro, and BAS1 displayed CDSP32-dependent peroxidase activity. The function of CDSP32 was investigated in transgenic potato lines without detectable levels of the protein as a result of cosuppression. Under conditions of photooxidative stress induced by incubation with either methyl viologen or t-butyl hydroperoxide or by exposure to low temperature under high light, plants lacking CDSP32 exhibited decreased maximal photosystem II photochemical efficiencies compared with the wild type and transgenic controls. In addition, plants without CDSP32 retained much less chlorophyll than controls under stress, indicating increased damage to photosynthetic membranes. We conclude that CDSP32 is a thioredoxin with a critical role in plastid defense against oxidative damage and that this role is related to its function as a physiological electron donor to the BAS1 peroxiredoxin. PMID- 12084839 TI - Perinatal events, vitamin D, and the development of allergy. PMID- 12084840 TI - Mode of delivery and asthma -- is there a connection? AB - Genetic factors cannot explain the recent rapid increase in the incidence of atopic diseases. The phenomenon has been explained by environmental factors, and there are data for and against the hypothesis that a decline in the pressure of microbial stimulation early in life could be behind the allergy epidemic. Changes have also occurred in maternity care, among them a rise in the caesarean section rate, which could diminish initial microbial exposure and thereby alter T helper 1 cell/T helper-2 cell development and affect the risk of developing atopy. In this study, we sought to establish whether mode of delivery does influence the development of atopic asthma. Finnish 1987 Medical Birth Register (n = 59,927 live births) information was linked between several national health registers to obtain information on asthma and mode of delivery in children registered. The data were adjusted for maternal age, previous deliveries, child's sex, and birth size. Atopy was evaluated in the second study (Turku Birth Cohort), which involved 219 children born by vaginal delivery (n = 106) or caesarean section (n = 113); history of atopic symptoms was established by questionnaire and a clinical examination was conducted, including skin prick testing and determination of total and allergen-specific IgE in serum. The register study showed the cumulative incidence of asthma at the age of seven to be significantly higher in children born by caesarean section (4.2%) than in those vaginally delivered (3.3%), the adjusted odds ratio (OR) for confounding variables being 1.21 (1.08-1.36), p < 0.01. In the second study, significantly more positive allergy tests were reported in questionnaires in the caesarean (22%) than in the vaginal delivery group (11%), OR 2.22 (1.06-4.64), p < 0.01, and a trend toward more positive skin prick reactions was documented at clinical examination; 41% versus 29%, OR 1.31 (0.65-2.65), p = 0.11. In conclusion, these results suggest that caesarean section delivery may be associated with an increased prevalence of atopic asthma. PMID- 12084841 TI - 1 alpha,25(OH)2D3 inhibits not only Th1 but also Th2 differentiation in human cord blood T cells. AB - Human naive CD4+ T helper (Th) and CD8+ cytotoxic (Tc) T cells, which only produce IL-2, may differentiate into Th1/Tc1- or Th2/Tc2-like lymphocytes, characterized by their cytokine production profile. 1 alpha,25-dihydroxyvitamin D3 (1 alpha, 25(OH)2D3) has been reported to inhibit Th1/Tc1-related, but increase Th2/Tc2-associated cytokines in T cells from adults. In industrialized countries, vitamin D supplementation for prevention of rickets is initiated within the first days of life and continued throughout the entire first year. Epidemiologic studies suggest an association of vitamin D exposure in newborns with the incidence of allergic diseases in later life. This study addresses the effects of 1 alpha, 25(OH)2D3 on Th1/Tc1 versus Th2/Tc2 differentiation in long term cell cultures of (naive) cord blood T lymphocytes. Our results show that in CD4+ as well as CD8+ cord blood cells, 1 alpha, 25(OH)2D3 inhibits not only IL-12 generated IFN-gamma production, but also suppresses IL-4 and IL-13 expression induced by IL-4. Thus, in cord blood 1 alpha, 25(OH)2D3 induces a T cell population without predominance of Th2 related cytokines. PMID- 12084842 TI - Sildenafil reverses O2 constriction of the rabbit ductus arteriosus by inhibiting type 5 phosphodiesterase and activating BK(Ca) channels. AB - Oxygen constriction causes functional closure of the ductus arteriosus (DA) at birth. Although DA closure is crucial for postnatal adaptation, patency of the DA is critical for survival of newborns with duct-dependent cardiac malformations. In these cases, DA patency is achieved by i.v. infusion of prostaglandin E1, which, though effective, is often associated with complications. We hypothesized that sildenafil, a specific phosphodiesterase type 5 inhibitor, is an effective DA vasodilator. In isolated DA rings from term (d 30) fetal rabbits, sildenafil (10(-6)-10(-4) M) and diethylamine NONOate (10(-7)-10(-5) M) induced dose dependent relaxation of oxygen-constricted DA (-52 +/- 4% and -51 +/- 6%, respectively) that was inhibited by the soluble guanylyl-cyclase inhibitor, 1H [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (5 x 10(-5) M). Sildenafil increased cyclic GMP levels. Iberiotoxin (200 nM), an inhibitor of calcium-sensitive potassium channels, decreased the vasodilatory effect of sildenafil and diethylamine NONOate (-30 +/- 2% and -27 +/- 4%, respectively). Oxygen inhibition of whole-cell K+ current and membrane depolarization were partially restored by sildenafil, and this was inhibited by iberiotoxin. Immunohistochemistry and immunoblotting confirmed the presence of phosphodiesterase type 5 and calcium sensitive potassium channels in the DA smooth muscle cells. This is the first study to demonstrate that sildenafil dilates the DA by increasing soluble guanylyl-cyclase-derived cGMP levels and thereby activating calcium-sensitive potassium channels, causing membrane hyperpolarization. Sildenafil, already approved for human usage, might be an alternative or a useful adjunct to prostaglandin E1 as a bridge to cardiac surgery. PMID- 12084843 TI - A role for platelet-derived growth factor-BB in rat postpneumonectomy compensatory lung growth. AB - Unilateral pneumonectomy leads to compensatory growth in the residual lung, the mediators of which are largely unknown. We hypothesized, based on its other known roles in lung cell growth, that platelet-derived growth factor (PDGF)-BB would be an essential mediator of postpneumonectomy compensatory lung growth. Left-sided pneumonectomies were performed on 21-d-old rats, for comparison with sham operated or unoperated control animals. Body weights were not different between groups. Right lung weights and DNA content were significantly increased (p < 0.05), compared with controls, by 10 d after pneumonectomy. The rate of DNA synthesis was maximal on d 5 postpneumonectomy. Total right lung PDGF-B mRNA and PDGF-BB protein increased after pneumonectomy, but were apparently tightly regulated, relative to total right lung beta-actin mRNA and protein content, respectively. However, PDGF-BB expression after pneumonectomy was apparently not purely constitutive, in that daily i.p. injections of a truncated soluble PDGF beta-receptor both reduced activation of the native PDGF beta-receptor, and attenuated increased lung DNA synthesis on d 3 after pneumonectomy. These findings are consistent with a critical role for PDGF-BB in postpneumonectomy lung growth. PMID- 12084844 TI - Effect of glutathione on lung activator protein-1 activation and heme oxygenase-1 induction in the immature rat. AB - Differences in lung heme oxygenase-1 (HO-1) regulation have been demonstrated in newborn (<12 h old) and adult (>2 month old) rats after exposure to hyperoxia. Contrary to adults, neonates do not demonstrate increased lung HO-1 induction nor transcription factor activator protein-1 (AP-1) binding in hyperoxia. Because AP 1 activation can be posttranslationally modified by oxidants or reductants, we investigated whether differences in lung glutathione (GSH) content account for the maturational differences in AP-1 activation and subsequent HO-1 gene regulation after hyperoxia. Neonatal rats were injected with either 1-buthionine [S,R] sulfoximine (BSO), diamide, or selenite during the 72-h hyperoxic exposure. Lung GSH content, glutathione disulfide (GSSG) content, AP-1 binding, and HO-1 mRNA were evaluated. The ratios of GSSG to GSH were used to reflect the GSH redox state in the lungs. Changes in lung GSSG/GSH ratio did not alter AP-1 binding but did increase HO-1 mRNA in neonates. These data suggest that the neonatal lung is relatively resistant to AP-1 activation and HO-1 induction by GSH perturbation. PMID- 12084846 TI - Oxidative stress in preterm neonates at birth and on the seventh day of life. AB - Previous studies have demonstrated increased oxidative damage to proteins and increased lipid peroxidation products in the plasma of hypoxic newborns at birth. We tested the hypothesis that hypoxic preterm newborns are at increased risk for oxidative stress in the first week of life. Heparinized blood samples of 34 hypoxic and 15 control preterm newborns were obtained at birth from the umbilical vein immediately after delivery and from a peripheral vein on postnatal d 7. Plasma levels of hypoxanthine, total hydroperoxide (TH), and advanced oxidation protein products (AOPP) were measured in cord blood and blood drawn on d 7. Hypoxanthine, TH, and AOPP levels were significantly higher in cord and d 7 blood samples of hypoxic newborn than control infants. Statistically significant correlations were observed between AOPP and hypoxanthine and between AOPP and TH plasma levels on d 7. AOPP and TH plasma levels significantly increased from cord to d 7 blood in neonates without hypoxia. These findings show that the oxidative stress observed in cord blood of hypoxic preterm newborns is still higher than control infants on d 7. The significant increase in TH and AOPP levels in nonhypoxic preterm newborns at the end of the first postnatal week indicates that damage caused by free radicals also occurs in nonhypoxic babies with normal clinical course. In summary, TH and AOPP production is prolonged for several days after birth in hypoxic preterm babies. The risk of free radical damage is lower but still exists in preterm neonates with normal clinical course. PMID- 12084845 TI - Biphasic ATP depletion caused by transient oxidative exposure is associated with apoptotic cell death in rat embryonal cortical neurons. AB - Hypoxia-ischemia leads to an acute depletion of high-energy phosphates in neonatal brain. After reperfusion, energy status is restored, but may show progressive secondary failure, associated with neuronal loss, brain damage, or death. Oxidants are produced on reperfusion. We investigated whether a biphasic energy failure develops in cultured neurons after oxidant exposure, and whether the degree of primary disturbance correlates with later ATP synthesis and mode of cell death. Embryonic rat cortical neurons were exposed to varying doses of hydrogen peroxide for 60 min and incubated for 12, 24, or 48 h. Adenine nucleotides and the incorporation of [(14)C]adenine into adenine nucleotides were quantified. Apoptosis was evaluated by DNA electrophoresis and in situ end labeling. A mild insult (10-50 microM) caused no ATP depletion or change in subsequent growth or energy metabolism, whereas an intermediate insult (100 microM) caused acute ATP depletion (49 +/- 12% of control). This recovered to 91 +/- 28% by 12 h, but then declined to 61 +/- 18% at 24 h. A severe insult (1 mM) depleted ATP to 15 +/- 3% of control, with no recovery. Moderate ATP depletion was associated with apoptotic cell death, whereas a severe insult caused acute necrosis. Transient oxidant exposure of embryonal cortical neurons causes a biphasic energy depletion followed by apoptosis in analogy with asphyxiated brains. This model may prove useful for the study of pathogenesis and treatment of hypoxic-ischemic encephalopathy. PMID- 12084847 TI - Effects of hypoxia on respiratory neural output and lower esophageal sphincter pressure in piglets. AB - We have previously documented anatomic and functional relationships between ventilatory and autonomic neural output. Therefore, we hypothesized in this study that hypoxia-induced changes in respiratory neural output are associated with changes in autonomic regulation of lower esophageal sphincter (LES) pressure. Respiratory neural output, heart rate, and LES pressure were measured before and during a 3-min exposure to 8% oxygen (balance nitrogen) in 12 3- to 7-d-old piglets. Respiratory neural output was determined from diaphragmatic electromyogram and LES pressure from an esophageal catheter. Studies were repeated after atropine administration in eight animals. Hypoxic exposure resulted in significant increases in diaphragmatic amplitude, respiratory rate, and minute diaphragmatic activity as well as heart rate. The biphasic response of diaphragm amplitude peaked at 1 min, whereas the responses of respiratory frequency and heart rate were sustained. Hypoxia caused a 50% increase in LES pressure (p < 0.05), which was eliminated by i.v. atropine administration. Development of apnea during subsequent hyperoxic exposure was always followed by a decline in LES pressure. Hypoxia-induced increase in respiratory neural output and accompanying increase in heart rate are associated with enhanced constrictive output to the LES. Blockade by atropine implicates a peripheral cholinergic mechanism for this LES response. We speculate that whereas hypoxia in the presence of enhanced respiratory neural output seems to be protective against reflux, decreased respiratory drive and accompanying apnea may be associated with a decline in LES tone and predispose to gastroesophageal reflux. PMID- 12084848 TI - 1H-NMR spectroscopy of cerebrospinal fluid of fetal sheep during hypoxia-induced acidemia and recovery. AB - The purpose of the study was to investigate the sequence of processes occurring during and after hypoxia-induced acidemia. We used proton nuclear magnetic resonance spectroscopy, which provides an overview of metabolites in cerebrospinal fluid (CSF), reflecting neuronal metabolism and damage. The pathophysiological condition of acute fetal asphyxia was mimicked by reducing maternal uterine blood flow in 14 unanesthetized pregnant ewes. CSF metabolites were measured during hypoxia-induced acidemia, and during the following recovery period, including the periods at 24 and 48 h after the hypoxic insult. Maximum values of the following CSF metabolites were reached during severe hypoxia (pH 85 and neuromotor scores (NMS) <3; n = 15], abnormal [if MDI or=3 at 12 and 30 mo; n = 11], or deteriorated [if normal at 12 mo and abnormal at 30 mo (MDI or=3); n = 11]. Thirty percent (11/37) of our cohort deteriorated between 12 and 30 mo. N-acetylaspartate/choline decreased across the groups ordered as normal, deteriorated, and abnormal [in basal nuclei (p 50 chromosomes) than in patients with diploid or hyperdiploid (46-50 chromosomes) leukemic blasts. PMID- 12084859 TI - Effects of hyperbilirubinemia on cerebrocortical electrical activity in newborns. AB - In our study, cerebrocortical electrical activity was recorded as an indicator of bilirubin neurotoxicity. Bilirubin especially affects the thalamus and cerebral cortex. Inasmuch as rhythmic oscillations on the EEG arise from the interaction between cortex and thalamus, electrophysiologic effects of bilirubin on the rhythmic oscillations with long-term postnatal age were investigated. Brain maturation was also analyzed with power spectral analysis quantitatively. For this purpose, 141 EEG records were taken (in the first week, 15th d, at the end of the first month, and at the third month) from 17 infants with hyperbilirubinemia and 22 healthy infants. In all records, the major frequency component was formed by the delta frequency in both groups. In the first records of the hyperbilirubinemia group, the delta frequency was higher than the control group; however the theta, alpha, and beta frequencies and the amplitude levels were lower (p < 0.001). These changes were found to be significantly correlated with the bilirubin levels (p < 0.001). On the 15th d the amplitude of the hyperbilirubinemia group increased to similar levels as the control group. At the frequency bands of delta and theta, there were significant changes related to postnatal age (p < 0.001). In all cerebral regions, the delta frequency decreased and the theta frequency increased with age. However, in the hyperbilirubinemia group the delta frequency was higher, the theta frequency was lower, and the changes between the groups disappeared in the third month despite the differences at all regions of the brain. In terms of the vertex, k complex, and sleep spindle, there were no differences between both groups in the third month (p > 0.05). We conclude that hyperbilirubinemia affects the cerebrocortical electrical activity but appears to be time limited. PMID- 12084858 TI - Palm versus soybean oil on intestinal recovery from malnutrition in Guinea pigs. AB - Recent recommendations on feeding malnourished children do not provide indication on the nature of dietary lipids. Our aim was to compare the effect of palm oil (mainly saturated and monounsaturated fatty acids) and soybean oil (mainly polyunsaturated fatty acids) on the recovery from malnutrition in guinea pigs. In a first experiment, guinea pigs received a balanced (control group) or a maize (malnourished group) diet for 7, 12, and 21 d. In a second experiment, after 12 d of malnutrition, guinea pigs received a rehabilitation diet containing palm or soybean oil. Both rehabilitation diets allowed a partial recovery from the severe weight loss induced by malnutrition. Thiobarbituric acid reactive substances content, measured in intestinal homogenates, increased in malnourished guinea pigs compared with control animals (40%, p < 0.05) and returned to near control values after rehabilitation with palm (10%) but not soybean (43%) oil diet. Intestinal short-circuit current, assessed in jejunal segments mounted in Ussing chambers, increased progressively during malnutrition (p < 0.001) and returned to near control values with both rehabilitation diets. Compared with control animals, the cell turnover (Ki-67 index assessed by immunohistochemistry detection of the Ki-67 antigen) decreased after soybean (-60%, p < 0.01) but not after palm oil. These results confirm that experimental polydeficient malnutrition induces oxidative stress and dysfunction in the intestine. They show a differential effect of palm and soybean oil on these intestinal measurements, suggesting that the composition of dietary lipids may be important in the treatment of malnutrition. PMID- 12084860 TI - New dosages and formulations of AEDs for use in pediatric epilepsy. PMID- 12084862 TI - Integrating clinical trial data into clinical practice. PMID- 12084863 TI - Epilepsy surgery in children: when, why, and how. PMID- 12084864 TI - The ketogenic diet: uses and abuses. PMID- 12084867 TI - In-flight neurologic emergencies. PMID- 12084868 TI - Time is money--or is it? Estimating the costs of informal caregiving. PMID- 12084869 TI - Practice parameter: neuroimaging of the neonate: report of the Quality Standards Subcommittee of the American Academy of Neurology and the Practice Committee of the Child Neurology Society. AB - OBJECTIVE: The authors reviewed available evidence on neonatal neuroimaging strategies for evaluating both very low birth weight preterm infants and encephalopathic term neonates. IMAGING FOR THE PRETERM NEONATE: Routine screening cranial ultrasonography (US) should be performed on all infants of <30 weeks' gestation once between 7 and 14 days of age and should be optimally repeated between 36 and 40 weeks' postmenstrual age. This strategy detects lesions such as intraventricular hemorrhage, which influences clinical care, and those such as periventricular leukomalacia and low-pressure ventriculomegaly, which provide information about long-term neurodevelopmental outcome. There is insufficient evidence for routine MRI of all very low birth weight preterm infants with abnormal results of cranial US. IMAGING FOR THE TERM INFANT: Noncontrast CT should be performed to detect hemorrhagic lesions in the encephalopathic term infant with a history of birth trauma, low hematocrit, or coagulopathy. If CT findings are inconclusive, MRI should be performed between days 2 and 8 to assess the location and extent of injury. The pattern of injury identified with conventional MRI may provide diagnostic and prognostic information for term infants with evidence of encephalopathy. In particular, basal ganglia and thalamic lesions detected by conventional MRI are associated with poor neurodevelopmental outcome. Diffusion-weighted imaging may allow earlier detection of these cerebral injuries. RECOMMENDATIONS: US plays an established role in the management of preterm neonates of <30 weeks' gestation. US also provides valuable prognostic information when the infant reaches 40 weeks' postmenstrual age. For encephalopathic term infants, early CT should be used to exclude hemorrhage; MRI should be performed later in the first postnatal week to establish the pattern of injury and predict neurologic outcome. PMID- 12084870 TI - Is there a neurologist on this flight? AB - OBJECTIVE: To analyze the frequency of neurologic events during commercial airline flights and to assess whether onboard emergency medical kits are adequate for in-flight neurologic emergencies. METHODS: Collaboration of the Mayo Clinic's Departments of Emergency Medicine and Medical Transportation Service and the Division of Aerospace Medicine to provide real-time in-flight consultation to a major US airline that flies approximately 10% of all US passengers. We analyzed all medical events reported from 1995 to 2000 in a database that catalogs the air to-ground medical consultations. All cases with potential neurologic symptoms were reviewed and classified into various neurologic symptom categories. The cost of diversion for each neurologic symptom was calculated and then extrapolated to assess the cost of neurologic symptoms to the US airline industry. RESULTS: A total of 2,042 medical incidents led to 312 diversions. Neurologic symptoms were the single largest category of medical incidents, prompting 626 air-to-ground medical calls (31%). They caused 34% of all diversions. Dizziness/vertigo was the most common neurologic symptom followed by seizures, headaches, pain, and cerebrovascular symptoms. Whereas seizures and dizziness/vertigo were the most common reasons for diversion, loss of consciousness/syncope was the complaint most likely to lead to a diversion. The estimated annual cost of diversions due to neurologic events is almost 9,000,000 dollars. CONCLUSION: Neurologic symptoms are the most common medical complaint requiring air-to-ground medical support and are second only to cardiovascular problems for emergency diversions and their resultant costs to the US airline industry. Adding antiepileptic drugs to the onboard medical kit and greater emergency medical training for in-flight personnel could potentially reduce the number of diversions for in-flight neurologic incidents. PMID- 12084871 TI - Comparative diagnostic utility of 1H MRS and DWI in evaluation of temporal lobe epilepsy. AB - OBJECTIVE: To compare the ability of diffusion-weighted MRI (DWI) and (1)H MRS to lateralize to the temporal lobe of seizure onset and to predict postoperative seizure control in patients with temporal lobe epilepsy (TLE). METHODS: Forty TLE patients who subsequently underwent epilepsy surgery and 20 normal subjects were studied with (1)H MRS and DWI. Medial parietal and temporal lobe N acetylaspartate (NAA)/creatine (Cr) ratios and hippocampal and temporal stem apparent diffusion coefficients (ADC) were obtained. Lateralization to either temporal lobe with each MR measurement was based on the threshold values derived from +/-1-SD right/left ratios of normal subjects. RESULTS: Temporal lobe NAA/Cr lateralized to the operated temporal lobe in 18 of 40 (45%), hippocampal ADC in 32 of 40 (80%), and temporal stem ADC in 26 of 40 (65%) patients. Almost all of the cases that lateralized to the surgical side with NAA/Cr ratios (94%) had an excellent postoperative seizure control (p = 0.01). Lateralization to the side of surgery was not associated with surgical outcome with hippocampal and temporal stem ADC (p > 0.05). CONCLUSION: (1)H MRS and DWI complement each other in the clinical setting. DWI more frequently lateralized to the operated side, and (1)HMRS was a better predictor of postoperative seizure control. PMID- 12084872 TI - A national study of the quantity and cost of informal caregiving for the elderly with stroke. AB - BACKGROUND: As the US population ages, increased stroke incidence will result in higher stroke-associated costs. Although estimates of direct costs exist, little information is available regarding informal caregiving costs for stroke patients. OBJECTIVE: To determine a nationally representative estimate of the quantity and cost of informal caregiving for stroke. METHODS: The authors used data from the first wave of the Asset and Health Dynamics (AHEAD) Study, a longitudinal study of people over 70, to determine average weekly hours of informal caregiving. Two part multivariable regression analyses were used to determine the likelihood of receiving informal care and the quantity of caregiving hours for those with stroke, after adjusting for important covariates. Average annual cost for informal caregiving was calculated. RESULTS: Of 7,443 respondents, 656 (8.8%) reported a history of stroke. Of those, 375 (57%) reported stroke-related health problems (SRHP). After adjusting for cormorbid conditions, potential caregiver networks, and sociodemographics, the proportion of persons receiving informal care increased with stroke severity, and there was an association of weekly caregiving hours with stroke +/- SRHP (p < 0.01). Using the median 1999 home health aide wage (8.20 dollars/hour) as the value for family caregiver time, the expected yearly caregiving cost per stroke ranged from 3,500 dollars to 8,200 dollars. Using conservative prevalence estimates from the AHEAD sample (750,000 US elderly patients with stroke but no SRHP and 1 million with stroke and SRHP), this would result in an annual cost of up to 6.1 billion dollars for stroke related informal caregiving in the United States. CONCLUSIONS: Informal caregiving-associated costs are substantial and should be considered when estimating the cost of stroke treatment. PMID- 12084873 TI - A multidrug combination designed for reversing resistance to BCNU in glioblastoma multiforme. AB - BACKGROUND: Nitrosoureas constitute the main resource of chemotherapy for glioblastoma. However, because of chemoresistance, which is intrinsic or rapidly acquired after the first administration of chemotherapy, there have been few improvements in survival. Because O(6)-alkylguanine-DNA alkyltransferase (AGT) is the main target for increasing cell sensitivity to the nitrosoureas, we postulated that preexposure to other alkylating agents might increase the therapeutic index of the nitrosoureas by saturating all the copies of AGT present in the tumor cells. OBJECTIVE: To investigate the response rate, toxic effects, time from start of chemotherapy to progression of disease or exit from the study for any reason (TTP), and progression-free survival at 6 months (PFS-6) associated with a multidrug combination that could reverse resistance to carmustine (BCNU) through AGT depletion. METHODS: We conducted a phase 2 study of patients with glioblastoma at first relapse or progression after surgery and standard radiotherapy. Patients were treated with 100 mg/m(2) of procarbazine on days 1 to 5, 80 mg/m(2) of BCNU on days 3 to 5, and 1.4 mg/m(2) of vincristine on day 3 every 8 weeks. RESULTS: Fifty-eight patients were enrolled in the study, and all were assessable for response and toxic effects. Six patients (10.3%) had a complete response, 11 (19%) had a partial response, and 17 (29.3%) had stable disease. The median TTP was 4.8 months; 42.3% of patients had PFS-6, and 15.4% had PFS at 12 months. Response to chemotherapy was the only significant prognostic factor for TTP. Neutropenia was grade 3 in 8.6% of patients and grade 4 in 5.2% of patients, and thrombocytopenia was grade 3 in 17.2% of patients and grade 4 in 12% of patients; hepatic and pulmonary toxic effects were grade 3 in 5.2% and 8.6% of patients, respectively. CONCLUSION: This regimen proved active in chemotherapy-naive patients with recurrent glioblastoma even though toxic effects were substantial. PMID- 12084875 TI - Hereditary neuropathy with liability to pressure palsy: the electrophysiology fits the name. AB - BACKGROUND: Studies of patients with hereditary neuropathy with liability to pressure palsies (HNPP) have shown accentuated distal slowing along with nonuniform conduction abnormalities at segments liable to compression, suggesting a distal myelinopathy as an underlying pathophysiological mechanism. METHODS: We evaluated 12 patients with HNPP by standard nerve conduction studies and by conduction to more proximal muscles in the arm and leg. Three CMT1A patients and six healthy subjects also were evaluated as controls. RESULTS: Median and peroneal motor nerves in all HNPP patients showed prolonged distal motor latencies (DML) (mean +/- SE, 5.9 +/- 0.41 and 8.63 +/- 0.58 milliseconds), but the ulnar and tibial DML were minimally prolonged or normal (mean +/- SE, 3.87 +/ 0.16 and 5.66 +/- 0.24 milliseconds). DML to forearm flexor (median and ulnar nerves) or anterior tibial muscles (peroneal nerve) were also normal. CONCLUSION: Accentuated distal slowing is found primarily in median and peroneal nerve segments liable to pressure palsies or repetitive trauma. However, the ulnar and tibial nerves, which are less liable to compression, have minimal changes. In addition, distal latencies to more proximal muscles in the arm and leg do not have distal slowing. These findings do not support a distal myelinopathy as a determinant of the conduction abnormalities in HNPP. PMID- 12084874 TI - Incidence of and risk factors for HIV-associated distal sensory polyneuropathy. AB - OBJECTIVE: To assess the incidence of and risk factors for distal sensory polyneuropathy (DSP) in a cohort of HIV-infected subjects. METHODS: We followed 272 subjects semiannually for up to 30 months. DSP was diagnosed if subjects had decreased or absent ankle jerks, decreased or absent vibratory perception at the toes, or decreased pinprick or temperature in a stocking distribution. Subjects were further classified at each visit as having asymptomatic DSP (ADSP) (signs only) or symptomatic DSP (SDSP) if, in addition to the neurologic signs, paresthesias or pain was reported. RESULTS: At baseline, 45% of the subjects did not meet criteria for DSP, 20% met criteria for ADSP, and 35% met criteria for SDSP. Dideoxynucleoside therapy was used by 23% of the patients, and this treatment was independent of their neuropathy status. In longitudinal univariate analyses, history of AIDS diagnoses (hazard ratio [HR] = 1.89; p = 0.02) and lower CD4 cell count (HR = 0.69; p = 0.0006) were risk factors for incident DSP (ADSP or SDSP). However, for incident SDSP only, in addition to history of AIDS diagnoses, mood and neurologic (other than DSP) and functional abnormalities were significant risk factors. Functional abnormalities remained a significant risk factor in a multiple regression analysis. The presence of ADSP and the use of dideoxynucleosides at baseline were not significant risk factors for incident SDSP. The Kaplan-Meier estimate of the 1-year incidence of SDSP was 36%. CONCLUSION: Subjects with moderate-to-severe immunosuppression from HIV infection commonly have SDSP. However, sex, use of dideoxynucleosides, and presence of ADSP were not significant risk factors for SDSP. PMID- 12084876 TI - Clinicopathological features of genetically confirmed Danon disease. AB - BACKGROUND: Danon disease is due to primary deficiency of lysosome-associated membrane protein-2. OBJECTIVE: To define the clinicopathologic features of Danon disease. METHODS: The features of 20 affected men and 18 affected women in 13 families with genetically confirmed Danon disease were reviewed. RESULTS: All patients had cardiomyopathy, 18 of 20 male patients (90%) and 6 of 18 female patients (33%) had skeletal myopathy, and 14 of 20 male patients (70%) and one of 18 female patients (6%) had mental retardation. Men were affected before age 20 years whereas most affected women developed cardiomyopathy in adulthood. Muscle histology revealed basophilic vacuoles that contain acid phosphatase-positive material within membranes that lack lysosome-associated membrane protein-2. Heart transplantation is the most effective treatment for the otherwise lethal cardiomyopathy. CONCLUSIONS: Danon disease is an X-linked dominant multisystem disorder affecting predominantly cardiac and skeletal muscles. PMID- 12084877 TI - Differential expression of chemokines in inflammatory myopathies. AB - BACKGROUND: Chemokines represent a family of small-molecular-weight cytokines that recruit and activate inflammatory cells in response to inflammation. Invasion of cytotoxic memory T cells and macrophages in nonnecrotic muscle fibers characterizes polymyositis and sporadic inclusion body myositis. Dermatomyositis is a complement-mediated endotheliopathy. Elucidation of the mechanisms guiding lymphocyte diapedesis and trafficking could lead to selective therapeutic interventions. METHODS: Immunoblots and multistep immunofluorescence studies with non-cross-reactive antibodies recognizing interleukin-8, monocyte chemoattractant protein-1 (MCP-1), MCP-3, TARC (thymus and activation regulated cytokine), and RANTES (regulated upon activation, normal T-cell expressed and secreted), using appropriate positive and negative controls. In situ hybridization was used to localize MCP-1 mRNA. RESULTS: MCP-1 protein was strongly expressed on T cells and a subset of macrophages actively invading a proportion of the nonnecrotic muscle fibers in polymyositis and inclusion body myositis alike. Capillaries and arterioles in the vicinity of endomysial inflammatory foci were immunoreactive for MCP-1, with faint or no expression in unaffected parts of the tissue. By contrast, widespread and strong endothelial MCP-1 expression occurred on perifascicular and perimysial endothelia in dermatomyositis, also at sites remote from inflammatory infiltrates. In some control specimens, a subset of capillaries also expressed MCP-1, possibly reflecting a role of this chemokine in normal immune surveillance. MCP-1 mRNA was detected in scattered macrophages in each inflammatory myopathy. All other chemokines were absent. CONCLUSION: Chemokines are differentially expressed in the symptomatic stage of inflammatory myopathies. MCP-1 plays a major role in the myocytotoxicity in polymyositis and inclusion body myositis. MCP-1 may be induced by membranolytic attack complex binding to endothelial cells in dermatomyositis. PMID- 12084878 TI - Neutralizing antibodies reduce MxA protein induction in interferon-beta-1a treated MS patients. AB - BACKGROUND: Neutralizing antibodies (NAb) during interferon-beta (IFNbeta) treatment of MS are associated with reduced clinical and MR efficacy. NAb inhibit the IFN- inducible MxA gene expression and neutralize the capability of IFNbeta to inhibit virus growth in vitro. Presently, there is no clear concept of the biologic importance of IFNbeta antibodies; most of the tests applied for the detection of NAb in previous publications are not widely available, and the results are not fully comparable. METHODS: A 1-year prospective study of the development of binding antibodies (BAb) and NAb and their relationship to IFN inducible MxA protein levels in peripheral blood leukocytes in 20 IFNbeta-1a treated patients with relapsing-remitting MS was conducted. RESULTS: In seven of nine NAb-positive patients, IFNbeta-1a was unable to induce MxA protein. BAb were detected in 11 patients, and they preceded or paralleled the development of NAb in all the patients. The titer of NAb correlated positively with BAb titer and negatively with MxA expression level. There was also a weaker but clear correlation between BAb titers and MxA levels. CONCLUSIONS: NAb, in most but not all cases, inhibited the in vivo function of IFNbeta. Analysis of MxA protein in lymphocytes together with analysis of NAb is a promising marker for evaluating the biologic effects of IFNbeta treatment in MS patients. PMID- 12084879 TI - Phases of A beta-deposition in the human brain and its relevance for the development of AD. AB - BACKGROUND: The deposition of the amyloid beta protein (Abeta) is a histopathologic hallmark of AD. The regions of the medial temporal lobe (MTL) are hierarchically involved in Abeta-deposition. OBJECTIVE: To clarify whether there is a hierarchical involvement of the regions of the entire brain as well and whether there are differences in the expansion of Abeta-pathology between clinically proven AD cases and nondemented cases with AD-related pathology, the authors investigated 47 brains from demented and nondemented patients with AD related pathology covering all phases of beta-amyloidosis in the MTL (AbetaMTL phases) and four control brains without any AD-related pathology. METHODS: Abeta deposits were detected by the use of the Campbell-Switzer silver technique and by immunohistochemistry in sections covering all brain regions and brainstem nuclei. It was analyzed how often distinct regions exhibited Abeta deposits. RESULTS: In the first of five phases in the evolution of beta-amyloidosis Abeta deposits are found exclusively in the neocortex. The second phase is characterized by the additional involvement of allocortical brain regions. In phase 3, diencephalic nuclei, the striatum, and the cholinergic nuclei of the basal forebrain exhibit Abeta deposits as well. Several brainstem nuclei become additionally involved in phase 4. Phase 5, finally, is characterized by cerebellar Abeta-deposition. The 17 clinically proven AD cases exhibit Abeta-phases 3, 4, or 5. The nine nondemented cases with AD-related Abeta pathology show Abeta-phases 1, 2, or 3. CONCLUSIONS: Abeta-deposition in the entire brain follows a distinct sequence in which the regions are hierarchically involved. Abeta-deposition, thereby, expands anterogradely into regions that receive neuronal projections from regions already exhibiting Abeta. There are also indications that clinically proven AD cases with full-blown beta-amyloidosis may be preceded in early stages by nondemented cases exhibiting AD-related Abeta pathology. PMID- 12084880 TI - Cognitive profiles differ in autopsy-confirmed frontotemporal dementia and AD. AB - BACKGROUND: Frontotemporal dementia (FTD) is currently distinguished from AD primarily on the basis of behavioral features because studies of cognition have shown negligible or inconsistent differences. However, the poor discriminability of cognitive measures may relate to reliance on imprecise clinically diagnosed groups. Therefore, a retrospective examination of neuropsychological test performance in autopsy-confirmed patients is warranted. OBJECTIVE: To compare the pattern of cognitive deficits exhibited by patients with autopsy-confirmed FTD and AD. METHODS: The profiles of cognitive deficits exhibited by patients with neuropathologic diagnosis of FTD (n = 14) or AD (n = 28) were compared. The Mattis Dementia Rating Scale (MDRS), letter and category fluency tests, Wechsler Intelligence Scale for Children-Revised block design test, Boston naming test, and clock drawing test were administered. RESULTS: Multivariate analysis of covariance controlling for age, education, and level of dementia revealed that patients with FTD performed significantly worse than patients with AD on letter and category fluency tests but significantly better on the MDRS memory subscale, block design test, and clock drawing test. A logistic regression model, validated in an independent clinical sample, used letter fluency, MDRS memory, and block design scores to correctly classify 91% of AD patients and 77% of FTD patients. CONCLUSIONS: A double dissociation in the pattern of cognitive deficits exhibited by FTD and AD patients was demonstrated. The FTD patients were more impaired than AD patients on word generation tasks (i.e., verbal fluency) that are sensitive to frontal lobe dysfunction but less impaired on tests of memory and visuospatial abilities sensitive to dysfunction of medial temporal and parietal association cortices. PMID- 12084882 TI - Progression of gait disorder and rigidity and risk of death in older persons. AB - BACKGROUND: Bradykinesia, gait disturbance, rigidity, and tremor are common motor signs in old age. All of these signs are associated with increased morbidity and mortality, but the extent to which they are progressive is unknown. METHODS: Study participants were 787 older Catholic clergy members without clinically diagnosed PD, related conditions, or dementia at baseline. They were evaluated annually for up to 7 years, with >95% follow-up participation by survivors. Evaluations included administration of a modified version of the motor portion of the Unified PD Rating Scale (UPDRS), from which previously established measures of the global UPDRS and four specific motor signs were derived. Scores represent the percent of the total possible UPDRS score obtained. RESULTS: At baseline, the global UPDRS score ranged from 0 to 36.3 (mean +/- SD, 7.3 +/- 6.4). It increased by an average of 0.69 unit per year during follow-up, with more rapid progression in older persons, but there was wide variability with no progression in 21% of subjects and annual increases of up to 8.23 units in the remaining 79%. Of 129 persons who died, 106 had follow-up UPDRS data. In a proportional hazards model, risk of death was associated with both the level of the global UPDRS score at baseline and the annual rate of progression (both p < 0.001). Overall, risk of death in subjects who had some worsening of the global UPDRS score was 2.93 times the rate among those without progression (95% CI, 1.32-6.50). Gait disorder/postural reflex impairment and rigidity worsened, but bradykinesia and tremor did not. Risk of death was associated with worsening of gait/posture but not with the other signs. CONCLUSION: Gait disorder and rigidity, as assessed with the modified UPDRS, are usually progressive in old age. Both the severity of the gait disorder and its rate of progression are strongly associated with risk of death. PMID- 12084881 TI - Oxidative damage and cytogenetic analysis in leukocytes of Parkinson's disease patients. AB - BACKGROUND: Postmortem studies suggest excessive free radical toxicity in the substantia nigra of patients with PD. Increased lipid peroxidation and oxidative DNA damage have been reported in the CNS. Markers of oxidative stress have been identified in the blood of patients with PD. OBJECTIVE: To assess the presence of spontaneous chromosome and primary or oxidative DNA damage in peripheral blood leukocytes of patients with untreated PD. METHODS: Patients with de novo PD (20) and control subjects (16), matched for age, sex, and smoking habits, underwent cytogenetic analysis using the human lymphocyte micronucleus assay coupled with the fluorescence in situ hybridization technique and the Comet assay. RESULTS: Compared with controls, patients with PD showed an increase in the incidence of spontaneous micronuclei (p < 0.001); single strand breaks (p < 0.001); and oxidized purine bases (p < 0.05). Fluorescence in situ hybridization analysis showed micronuclei harboring acentric fragments. CONCLUSIONS: There is chromosomal, primary DNA damage and oxidative DNA damage demonstrable in lymphocytes of patients with untreated PD. PMID- 12084883 TI - Novel three-stage ascertainment method: prevalence of PD and parkinsonism in South Tyrol, Italy. AB - BACKGROUND: A number of community-based studies on the prevalence of PD have been conducted worldwide, but they are often extremely costly and time consuming. OBJECTIVE: To assess the prevalence of PD and parkinsonism for the population aged between 60 and 85 years in South Tyrol, Northern Italy, using a novel population-based three-stage ascertainment method. METHODS: Seven hundred fifty persons aged 60 to 85 years from South Tyrol received a validated screening mail questionnaire for parkinsonism. In the second stage of the ascertainment method, trained primary care physicians (PCP) identified all persons with possible parkinsonism among those screened positive. In the third stage, movement disorders specialists excluded or confirmed the diagnosis in all identified people. RESULTS: The response rate was 87.6%. The prevalence rate per 100 population over 65 years of age was 1.5 (95% CI 0.6 to 2.3) for PD and 2.2 (95% CI 1.2 to 3.3) for parkinsonism after having been adjusted to the 1991 European standard population. Overall, 78% (95% CI 59 to 96%) of patients with parkinsonism were newly detected through the survey. CONCLUSIONS: The prevalence of PD and parkinsonism in people aged over 65 in South Tyrol was similar to that observed in door-to-door surveys in other European countries. The novel three stage case ascertainment method employed proved a useful tool to substitute for expensive door-to-door surveys for prevalence studies of parkinsonism, detecting a high number of undiagnosed cases, particularly in geographically remote areas. PMID- 12084884 TI - Progressive multifocal leukoencephalopathy. PMID- 12084885 TI - Prevalence of narcolepsy symptomatology and diagnosis in the European general population. AB - OBJECTIVE: To determine the prevalence of narcolepsy in the general population of five European countries (target population 205,890,882 inhabitants). METHODS: Overall, 18,980 randomly selected subjects were interviewed (participation rate 80.4%). These subjects were representative of the general population of the UK, Germany, Italy, Portugal, and Spain. They were interviewed by telephone using the Sleep-EVAL expert system, which provided narcolepsy diagnosis according to the International Classification of Sleep Disorders (ICSD). RESULTS: Excessive daytime sleepiness was reported by 15% of the sample, with a higher prevalence in the UK and Germany. Napping two times or more in the same day was reported by 1.6% of the sample, with a significantly higher rate in Germany. Cataplexy (episodes of loss of muscle function related to a strong emotion), a cardinal symptom of narcolepsy, was found in 1.6% of the sample. An ICSD narcolepsy diagnosis was found in 0.047% of the sample: The narcolepsy was severe for 0.026% of the sample and moderate in 0.021%. CONCLUSION: This is the first epidemiologic study that estimates the prevalence of narcolepsy in the general population of these five European countries. The disorder affects 47 individuals/100,000 inhabitants. PMID- 12084886 TI - Influence of final search direction on tactile line bisection in normal subjects. AB - BACKGROUND: Many variables influence the tactile bisection performance of normal subjects. Of these, studies that investigated the starting point effect have reported inconsistent results. OBJECTIVE: To determine if the final search direction rather than the initial search direction (starting point) may have an effect on tactile bisection performance, especially when subjects are making multiple searches. METHODS: The authors divided the experiment into single- and multiple-search tasks. Thirty-two blindfolded normal subjects were asked to indicate the midpoint of a rod after a one-way search (single-search task) or after two or more one-way searches (multiple-search task). In each task, the subject started on either end of the horizontally placed rods. The same procedure was also conducted with rods oriented vertically or radially. Therefore, there were six conditions (two starting points x three orientations). RESULTS: In the single-search task, there was a significant effect of starting point or movement direction in all six conditions, with biases occurring toward the starting point from the true midpoint (this is termed the "overshoot phenomenon"). In the multiple-search task, however, there was no significant effect of the starting point in all six conditions. Rather, biases depended on the final movement direction in five of the six conditions, occurring toward the starting point of the final search. CONCLUSION: Future research is needed to understand the mechanism of the overshoot phenomenon in tactile bisection by normal subjects and how the overshoot phenomenon influences patients with neglect. PMID- 12084887 TI - Neuropathology of a case of dopa-responsive dystonia associated with a new genetic locus, DYT14. AB - Detailed autopsy findings are reported for a patient with dopa-responsive dystonia genetically related to the dopa-responsive dystonia locus DYT14 on chromosome 14q13. Substantia nigra and locus ceruleus showed a normal abundance of severely hypomelanized dopaminergic neurons and no Lewy body. In the nigra, the reduction of melanin pigment was found to be asymmetric between the two sides and uneven within neurons, and the lateral aspect of the nigra appeared more affected than the medial, in a pattern similar to the neuronal loss in PD. Dopa responsive dystonia has a unique neuropathologic signature that seems to be independent of its genotype. PMID- 12084888 TI - Botulinum toxin improves lid opening delays in blepharospasm-associated apraxia of lid opening. AB - Lid movement and EMG of the orbicularis oculi (OOc) were analyzed in 10 patients with apraxia of lid opening associated with blepharospasm before and after botulinum toxin treatment. The latencies to onset and to complete the eye opening and the time during which eye opening was sustained were studied in relation to OOc activity and compared with control values obtained in 12 healthy subjects. Following treatment there was an improvement of all lid opening measurements, a decrease of the abnormally prolonged OOc activity, and a reduction of the functional disability. PMID- 12084889 TI - Sudden coma from acute bilateral internal carotid artery territory infarction. AB - Six patients with bilateral internal carotid artery occlusion who presented with sudden loss of consciousness, quadriplegia, and initially intact brainstem reflexes are described. They soon lost brainstem reflexes and died within 3 days. The presumed causes of internal carotid artery occlusion were atherothrombosis in three patients and cardiogenic embolism in the others. This catastrophic stroke syndrome mimics severe brainstem stroke and has an extremely poor prognosis. PMID- 12084890 TI - History of head trauma and risk of intracranial meningioma: population-based case control study. AB - A population-based case-control study in western Washington state was performed to assess the relation between head trauma and meningioma. Based on 200 case and 400 control subjects, head trauma was associated with an increased risk of meningioma (odds ratio = 1.83; 95% CI = 1.28, 2.62), especially head traumas occurring 10 to 19 years before reference date (odds ratio = 4.33; 95% CI = 2.06, 9.10). A dose-response relationship was present for number, but not severity, of head traumas. Whether the associations observed in this study are causal remains unclear. PMID- 12084891 TI - Reduced cortical gray matter in narcolepsy: preliminary findings with voxel-based morphometry. AB - There is a selective loss of hypocretin/orexin-containing hypothalamic neurons in patients with narcolepsy. The authors compared MRI-derived gray matter maps of 12 patients with narcolepsy with matched controls using voxel-based morphometry to ascertain whether there are other structural brain abnormalities. Patients with narcolepsy showed bilateral cortical gray matter reductions predominantly in inferior temporal and inferior frontal brain regions. Relative global gray matter loss was independent of disease duration or medication history. No significant subcortical gray matter alterations were noted. PMID- 12084892 TI - High-dose cyclophosphamide without stem-cell rescue for refractory CIDP. AB - Four patients with chronic inflammatory demyelinating polyneuropathy (CIDP) who were refractory to conventional treatment were treated with high-dose cyclophosphamide (200 mg/kg over 4 days). All improved in functional status and muscle strength. Nerve conduction studies improved in three of four. Other immunomodulatory medications have been discontinued. High-dose cyclophosphamide can be given safely to patients with CIDP and patients with disease persistence after standard therapy may have a response that lasts for over 3 years and results in long-term disease remission. PMID- 12084893 TI - Spinal catheter insertion via seated lumbar puncture using a massage chair. PMID- 12084894 TI - Can a critical lower threshold of cerebral perfusion be determined? A pilot observational study. PMID- 12084895 TI - NDUFA-1 is not a nuclear modifier gene in Leber hereditary optic neuropathy. PMID- 12084896 TI - MNGIE: diarrhea and leukoencephalopathy. PMID- 12084897 TI - Daytime sleepiness and other sleep disorders in Parkinson's disease. PMID- 12084898 TI - Experience, competency, and education: graduating neurology residents' experience with tPA. PMID- 12084899 TI - Driving restrictions and people with epilepsy. PMID- 12084900 TI - Procerus sign in progressive supranuclear palsy. PMID- 12084901 TI - The cerebrospinal fluid production rate is reduced in dementia of the Alzheimer's type. PMID- 12084902 TI - The prevalence of neuropsychiatric syndromes in systemic lupus erythematosus. PMID- 12084903 TI - Liver and thyroid function and autoimmunity during interferon-beta 1b treatment for multiple sclerosis. PMID- 12084904 TI - Reproductive biology. PMID- 12084905 TI - Estrogen actions in the brain. AB - Understanding of the mechanisms of estrogen action in the brain has always been poor. Neurons in several brain regions do not harbor estrogen receptor alpha (ERalpha) and yet are estrogen responsive. It was formerly thought that these responses represented indirect actions of estrogen. It is now evident that these neurons express ERbeta and that estrogen receptors have diverse actions in the central nervous system. By clear delineation of the cellular expression and function of the two estrogen receptors, it is likely that, in the future, selective ERalpha and ERbeta ligands will be developed and used for treatment of depression and behavioral disorders and may be useful in preventing degenerative diseases, such as Alzheimer's and Parkinson's disease. PMID- 12084906 TI - Rapid actions of steroid receptors in cellular signaling pathways. AB - Steroid hormones regulate cellular processes by binding to intracellular receptors that, in turn, interact with discrete nucleotide sequences to alter gene expression. Because most steroid receptors in target cells are located in the cytoplasm, they need to get into the nucleus to alter gene expression. This process typically takes at least 30 to 60 minutes. In contrast, other regulatory actions of steroid hormones are manifested within seconds to a few minutes. These time periods are far too rapid to be due to changes at the genomic level and are therefore termed nongenomic or rapid actions, to distinguish them from the classical steroid hormone action of regulation of gene expression. The rapid effects of steroid hormones are manifold, ranging from activation of mitogen activated protein kinases (MAPKs), adenylyl cyclase (AC), protein kinase C (PKC), and heterotrimeric guanosine triphosphate-binding proteins (G proteins). In some cases, these rapid actions of steroids are mediated through the classical steroid receptor that can also function as a ligand-activated transcription factor, whereas in other instances the evidence suggests that these rapid actions do not involve the classical steroid receptors. One candidate target for the nonclassical receptor-mediated effects are G protein-coupled receptors (GPCRs), which activate several signal transduction pathways. One characteristic of responses that are not mediated by the classical steroid receptors is insensitivity to steroid antagonists, which has contributed to the notion that a new class of steroid receptors may be responsible for part of the rapid action of steroids. Evidence suggests that the classical steroid receptors can be localized at the plasma membrane, where they may trigger a chain of reactions previously attributed only to growth factors. Identification of interaction domains on the classical steroid receptors involved in the rapid effects, and separation of this function from the genomic action of these receptors, should pave the way to a better understanding of the rapid action of steroid hormones. PMID- 12084907 TI - A new type of photoreceptor in algae. PMID- 12084908 TI - Might a vanguard of mRNAs prepare cells for the arrival of herpes simplex virus? PMID- 12084909 TI - Omnipotent decoding potential resides in eukaryotic translation termination factor eRF1 of variant-code organisms and is modulated by the interactions of amino acid sequences within domain 1. AB - In eukaryotes, a single translational release factor, eRF1, deciphers three stop codons, although its decoding mechanism remains puzzling. In the ciliate Tetrahymena thermophila, UAA and UAG codons are reassigned to Gln codons. A yeast eRF1-domain swap containing Tetrahymena domain 1 responded only to UGA in vitro and failed to complement a defect in yeast eRF1 in vivo at 37 degrees C. This finding demonstrates that decoding specificity of eRF1 from variant code organisms resides at domain 1. However, the wild-type eRF1 hybrid fully restored the growth of eRF1-deficient yeast at 30 degrees C. Tetrahymena eRF1 contains a variant sequence, KATNIKD, at the tip of domain 1. The TASNIKD variant of hybrid eRF1 rendered the eRF1-nullified yeast viable, although in an in vitro assay, the same hybrid eRF1 responded only to UGA. Nevertheless, the yeast eRF1 bearing the KATNIKD motif instead of the TASNIKS heptapeptide present in higher eukaryotes remains omnipotent in vivo. Collectively, these data suggest that variant genetic code organisms like Tetrahymena have an intrinsic potential to decode three stop codons in vivo, and that interaction within domain 1 between the KAT tripeptide and other sequences modulates the decoding specificity of Tetrahymena eRF1. PMID- 12084910 TI - TRILOGY: Discovery of sequence-structure patterns across diverse proteins. AB - We describe a new computer program, trilogy, for the automated discovery of sequence-structure patterns in proteins. trilogy implements a pattern discovery algorithm that begins with an exhaustive analysis of flexible three-residue patterns; a subset of these patterns are selected as seeds for an extension process in which longer patterns are identified. A key feature of the method is explicit treatment of both the sequence and structure components of these motifs: each trilogy pattern is a pair consisting of a sequence pattern and a structure pattern. Matches to both these component patterns are identified independently, allowing the program to assign a significance score to each sequence-structure pattern that assesses the degree of correlation between the corresponding sequence and structure motifs. trilogy identifies several thousand high-scoring patterns that occur across protein families. These include both previously identified and potentially novel motifs. We expect that these sequence-structure patterns will be useful in predicting protein structure from sequence, annotating newly determined protein structures, and identifying novel motifs of potential functional or structural significance. Further details on 7,768 significant patterns identified by trilogy can be found at http://theory.lcs.mit.edu/trilogy. PMID- 12084911 TI - The rate-limiting step in the folding of a large ribozyme without kinetic traps. AB - A fundamental question in RNA folding is the nature of the rate-limiting step. Folding of large RNAs often is trapped by the need to undo misfolded structures, which precludes the study of the other, potentially more interesting aspects in the rate-limiting step, such as conformational search, metal ion binding, and the role of productive intermediates. The catalytic domain of the Bacillus subtilis RNase P RNA folds without a kinetic trap, thereby providing an ideal system to elucidate these steps. We analyzed the folding kinetics by using fluorescence and absorbance spectroscopies, catalytic activity, and synchrotron small-angle x-ray scattering. Folding begins with the rapid formation of early intermediates wherein the majority of conformational search occurs, followed by the slower formation of subsequent intermediates. Before the rate-limiting step, more than 98% of the total structure has formed. The rate-limiting step is a small-scale structural rearrangement involving prebound metal ions. PMID- 12084912 TI - Specificity and affinity motifs for Grb2 SH2-ligand interactions. AB - Protein-protein interactions are often mediated by the recognition of short continuous amino acid stretches on target proteins by specific binding domains. Affinity-based selection strategies have successfully been used to define recognition motifs for a large series of such protein domains. However, in many biological systems specificity of interaction may be of equal or greater importance than affinity. To address this issue we have developed a peptide library screening technology that can be used to directly define ligands for protein domains based on both affinity and specificity of interaction. We demonstrate the value of this approach by the selection of peptide ligands that are either highly specific for the Grb2 Src homology 2 (SH2) domain or that are cross-reactive between a group of related SH2 domains. Examination of previously identified physiological ligands for the Grb2 SH2 domain suggests that for these ligands regulation of the specificity of interaction may be an important factor for in vivo ligand selection. PMID- 12084913 TI - Isolation of receptor-ligand pairs by capture of long-lived multivalent interaction complexes. AB - We have combined phage display and array screening for the rapid isolation of pairs of interacting polypeptides. Our strategy, named SAC (selection by avidity capture), is based on the avidity effect, the formation of highly stable complexes formed by multivalent interactions; in our case, between a receptor (multivalently displayed on phage) and a ligand (coexpressed as a multimeric fusion protein). Capture of the long-lived interaction complex allows the isolation of phage bearing cognate interaction pairs, as we demonstrate for a range of interactions, including Ab-antigen pairs and the rapamycin-dependent interaction of FKBP-12 and FRAP. Cognate phage are enriched by SAC up to 1000 fold and interacting pairs can be identified by array screening. Application of SAC to Ab-antigen interactions as a model system yielded over 140 specific Abs to a single antigen and 92 Abs to three different fetal human brain antigens in a single round of SAC each. Our results suggest that SAC should prove useful for the identification and study of receptor-ligand interactions in particular among extracellular proteins, as well as for the rapid generation of specific Abs to multiple antigens. PMID- 12084914 TI - Autoregulation of a bacterial sigma factor explored by using segmental isotopic labeling and NMR. AB - Bacterial sigma factors combine with the catalytic core RNA polymerase to direct the process of transcription initiation through sequence-specific interactions with the -10 and -35 elements of promoter DNA. In the absence of core RNA polymerase, the DNA-binding function of sigma is autoinhibited by its own N terminal 90 amino acids (region 1.1), putatively by a direct interaction with conserved region 4.2, which binds the -35 promoter element. In the present work, this mechanism of autoinhibition was studied by using a combination of NMR spectroscopy and segmental isotopic labeling of a sigma70-like subunit from Thermotoga maritima. Our data argue strongly against a high-affinity interaction between these two domains. Instead we suggest that autoinhibition of DNA binding occurs through an indirect steric and/or electrostatic mechanism. More generally, the present work illustrates the power of segmental isotopic labeling for probing molecular interactions in large proteins by NMR. PMID- 12084915 TI - Interactions of mutant and wild-type flap endonucleases with oligonucleotide substrates suggest an alternative model of DNA binding. AB - Previous structural studies on native T5 5' nuclease, a member of the flap endonuclease family of structure-specific nucleases, demonstrated that this enzyme possesses an unusual helical arch mounted on the enzyme's active site. Based on this structure, the protein's surface charge distribution, and biochemical analyses, a model of DNA binding was proposed in which single stranded DNA threads through the archway. We investigated the kinetic and substrate-binding characteristics of wild-type and mutant nucleases in relation to the proposed model. Five basic residues R33, K215, K241, R172, and R216, are all implicated in binding branched DNA substrates. All these residues except R172 are involved in binding to duplex DNA carrying a 5' overhang. Replacement of either K215 or R216 with a neutral amino acid did not alter kcat appreciably. However, these mutant nucleases displayed significantly increased values for Kd and Km. A comparison of flap endonuclease binding to pseudoY substrates and duplexes with a single-stranded 5' overhang suggests a better model for 5' nuclease-DNA binding. We propose a major revision to the binding model consistent with these biophysical data. PMID- 12084916 TI - Gene activation by interaction of an inhibitor with a cytoplasmic signaling protein. AB - Galactose-inducible genes (GAL genes) in yeast Saccharomyces cerevisiae are efficiently transcribed only when the sequence-specific transcription activator Gal4p is activated. Activation of Gal4p requires the interaction between the Gal4p inhibitory protein Gal80p and the galactokinase paralog, Gal3p. It has been proposed that Gal3p binds to a Gal80p-Gal4p complex in the nucleus to activate Gal4p. Here, we present evidence that the Gal3p-Gal80p interaction occurs in the cytoplasm, and concurrently, Gal80p is removed from Gal4p at the GAL gene promoter. We also show that GAL gene expression can be activated by heterologous protein-protein interaction in the cytoplasm that is independent of galactose and Gal3p function. These results indicate that galactose-triggered Gal3p-Gal80p association in the cytoplasm activates Gal4p in the nucleus. PMID- 12084917 TI - Sequence determinants of the energetics of folding of a transmembrane four-helix bundle protein. AB - Although previous studies are beginning to point to the specific types of helix helix interactions that stabilize the folds of membrane-bound helical proteins, quantitative thermodynamic data on natural membrane proteins has been very limited. Here the database is expanded substantially by adding thermodynamic data for a series of sequence variants of M2 protein from influenza A virus. The M2 protein has a single transmembrane helix that homotetramerizes to form proton selective channels that are essential to virus function. To determine the contributions of specific residues to the folding of this protein, a series of transmembrane peptides with single-site changes near the core of the protein were studied by using sedimentation equilibrium analytical ultracentrifugation. Remarkably, a large number of the mutations increased the stability of the protein. The free energies of tetramerization of the variants can be understood in terms of current models for the structure of the protein. In general, the energetic consequences of the mutations are smaller than those observed for similar mutations in water-soluble proteins. This observation is consistent with previous studies and hence may represent a general phenomenon. PMID- 12084918 TI - Characterization of an orphan G protein-coupled receptor localized in the dorsal root ganglia reveals adenine as a signaling molecule. AB - The cloning of novel G protein-coupled receptors and the search for their natural ligands, a process called reverse pharmacology, is an excellent opportunity to discover novel hormones and neurotransmitters. Based on a degenerate primer approach we have cloned a G protein-coupled receptor whose mRNA expression profile indicates highest expression in the dorsal root ganglia, specifically in the subset of small neurons, suggesting a role in nociception. In addition, moderate expression was found in lung, hypothalamus, peripheral blood leukocytes, and ovaries. Guided by a receptor-activation bioassay, we identified adenine as the endogenous ligand, which activated the receptor potently and with high structural stringency. Therefore, we propose to name this receptor as the adenine receptor. Hormonal functions have already been demonstrated for adenine derivatives like 6-benzylaminopurine in plants and 1-methyladenine in lower animals. Here, we demonstrate that adenine functions as a signaling molecule in mammals. This finding adds a third family besides P1 and P2 receptors to the class of purinergic receptors. PMID- 12084919 TI - Hsp90 enables Ctf13p/Skp1p to nucleate the budding yeast kinetochore. AB - Binding of CBF3, a protein complex consisting of Ndc10p, Cep3p, Ctf13p, and Skp1p, to the centromere DNA nucleates kinetochore formation in budding yeast. Here, we investigate how the Ctf13p/Skp1p complex becomes competent to form the CBF3-centromere DNA complex. As revealed by mass spectrometry, Ctf13p and Skp1p carry two and four phosphate groups, respectively. Complete dephosphorylation of Ctf13p and Skp1p does not interfere with the formation of CBF3-centromere DNA complexes in vitro. Furthermore, deletion of corresponding phosphorylation sites results in viable cells. Thus, in contrast to the current view, phosphorylation of Ctf13p and Skp1p is not essential for the formation of CBF3-centromere DNA complexes. Instead, the formation of active Ctf13p/Skp1p requires Hsp90. Several lines of evidence support this conclusion: activation of heterologous Ctf13p/Skp1p by reticulocyte lysate is inhibited by geldanamycin and Hsp90 depletion. skp1 mutants exhibit growth defects on media containing geldanamycin. A skp1 mutation together with Hsp90 mutations exhibits synthetic lethality. An Hsp90 mutant contains decreased levels of active Ctf13p/Skp1p. PMID- 12084920 TI - A target essential for the activity of a nonacidic yeast transcriptional activator. AB - P201 is a short (eight-residue) nonacidic peptide that comprises a strong transcriptional activating region when tethered to DNA in yeast. Here we identify the mediator protein Gal11 as an essential target of P201. Deletion of Gal11, which modestly decreases activation elicited by the typical acidic yeast activator, abolishes activation by DNA-tethered P201. A point mutation in Gal11, which has no effect on other Gal11 functions, also greatly diminishes activation by DNA-tethered P201. P201 binds to a fragment of Gal11 in vivo and in vitro, and the interaction is diminished by mutations in either component that decrease activation in vivo. P201, unlike the typical yeast acidic activating region, does not work in mammalian cells, which is consistent with the notion that the unique target of P201 (Gal11) is absent from mammalian cells. PMID- 12084921 TI - Specific zinc-finger architecture required for HIV-1 nucleocapsid protein's nucleic acid chaperone function. AB - The nucleocapsid protein (NC) of HIV type 1 (HIV-1) is a nucleic acid chaperone that facilitates the rearrangement of nucleic acid secondary structure during reverse transcription. HIV-1 NC contains two CCHC-type zinc binding domains. Here, we use optical tweezers to stretch single lambda-DNA molecules through the helix-to-coil transition in the presence of wild-type and several mutant forms of HIV-1 NC with altered zinc-finger domains. Although all forms of NC lowered the cooperativity of the DNA helix-coil transition, subtle changes in the zinc-finger structures reduced NC's effect on the transition. The change in cooperativity of the DNA helix-coil transition correlates strongly with in vitro nucleic acid chaperone activity measurements and in vivo HIV-1 replication studies using the same NC mutants. Moreover, Moloney murine leukemia virus NC, which contains a single zinc finger, had little effect on transition cooperativity. These results suggest that a specific two-zinc-finger architecture is required to destabilize nucleic acids for optimal chaperone activity during reverse transcription in complex retroviruses such as HIV-1. PMID- 12084922 TI - How to describe protein motion without amino acid sequence and atomic coordinates. AB - This paper reports a computational method, the quantized elastic deformational model, that can reliably describe the conformational flexibility of a protein in the absence of the amino acid sequence and atomic coordinates. The essence of this method lies in the fact that, in modeling the functionally important conformational changes such as domain movements, it is possible to abandon the traditional concepts of protein structure (bonds, angles, dihedrals, etc.) and treat the protein as an elastic object. The shape and mass distribution of the object are described by the electron density maps, at various resolutions, from methods such as x-ray diffraction or cryo-electron microscopy. The amplitudes and directionality of the elastic deformational modes of a protein, whose patterns match the biologically relevant conformational changes, can then be derived solely based on the electron density map. The method yields an accurate description of protein dynamics over a wide range of resolutions even as low as 15-20 A at which there is nearly no visually distinguishable internal structures. Therefore, this method dramatically enhances the capability of studying protein motions in structural biology. It is also expected to have ample applications in related fields such as bioinformatics, structural genomics, and proteomics, in which one's ability to extract functional information from the not-so-well defined structural models is vitally important. PMID- 12084923 TI - Using deeply trapped intermediates to map the cytochrome c folding landscape. AB - Replacement of iron with cobalt(III) selectively introduces a deep trap in the folding-energy landscape of the heme protein cytochrome c. Remarkably, neither the protein structure nor the folding thermodynamics is perturbed by this metal ion substitution, as shown by data from spectroscopic and x-ray diffraction experiments. Through kinetics measurements, we have found parallel folding pathways involving several different misligated Co(III) species, and, as these folding intermediates persist for several hours under certain conditions, we have been able to elucidate fully their spectroscopic properties. The results, along with an analysis of the fluorescence energy-transfer kinetics during refolding, show that rapidly equilibrating populations of compact and extended polypeptide conformations are present until all molecules have reached the native structure. These measurements provide direct evidence that collapsed denatured structures are not substantially more stable than extended conformations of cytochrome c. PMID- 12084924 TI - Topological determinants of protein folding. AB - The folding of many small proteins is kinetically a two-state process that represents overcoming the major free-energy barrier. A kinetic characteristic of a conformation, its probability to descend to the native state domain in the amount of time that represents a small fraction of total folding time, has been introduced to determine to which side of the free-energy barrier a conformation belongs. However, which features make a protein conformation on the folding pathway become committed to rapidly descending to the native state has been a mystery. Using two small, well characterized proteins, CI2 and C-Src SH3, we show how topological properties of protein conformations determine their kinetic ability to fold. We use a macroscopic measure of the protein contact network topology, the average graph connectivity, by constructing graphs that are based on the geometry of protein conformations. We find that the average connectivity is higher for conformations with a high folding probability than for those with a high probability to unfold. Other macroscopic measures of protein structural and energetic properties such as radius of gyration, rms distance, solvent-accessible surface area, contact order, and potential energy fail to serve as predictors of the probability of a given conformation to fold. PMID- 12084925 TI - Binding sites of different geometries for the 16-3 phage repressor. AB - Prokaryotic repressor-operator systems provide exemplars for the sequence specific interactions between DNA and protein. The crucial atomic contacts of the two macromolecules are attained in a compact, geometrically defined structure of the DNA-protein complex. The pitch of the DNA interface seems an especially sensitive part of this architecture because changes in its length introduce new spacing and rotational relations in one step. We discovered a natural system that may serve as a model for investigating this problem: the repressor of the 16-3 phage of Rhizobium meliloti (helix-turn-helix class protein) possesses inherent ability to accommodate to various DNA twistings. It binds the cognate operators, which are 5'-ACAA-4 bp-TTGT-3' (O(L)) and 5'-ACAA-6 bp-TTGT-3' (O(R)) and thus differ 2 bp in length, and consequently the two half-sites will be rotated with respect to each other by 72 degrees in the idealized B-DNA (64 degrees by dinucleotide steps calculations). Furthermore, a synthetic intermediate (DNA sequence) 5'-ACAA-5 bp-TTGT-3' (O5) also binds specifically the repressor. The natural operators and bound repressors can form higher order DNA-protein complexes and perform efficient repression, whereas the synthetic operator repressor complex cannot do either. The natural operators are bent when complexed with the repressor, whereas the O5 operator does not show bending in electrophoretic mobility assay. Possible structures of the complexes are discussed. PMID- 12084926 TI - Alternate interactions define the binding of peptides to the MHC molecule IA(b). AB - We have solved the crystal structure of the MHCII molecule, IA(b), containing an antigenic variant of the major IA(b)-binding peptide derived from the MHCII IEalpha chain. The four MHC pockets at p1, p4, p6, and p9 that usually bind peptide side chains are largely empty because of alanines in the peptide at these positions. The complex is nevertheless very stable, apparently because of unique alternate interactions between the IA(b) and peptide. In particular, there are multiple additional hydrogen bonds between the N-terminal end of the peptide and the IA(b) alpha chain and an extensive hydrogen bond network involving an asparagine at p7 position of the peptide and the IA(b) beta chain. By using knowledge of the shape and size of the traditional side chain binding pockets and the additional possible interactions, an IA(b) peptide-binding motif can be deduced that agrees well with the sequences of known IA(b)-binding peptides. PMID- 12084927 TI - CD25+CD4+ T cells contribute to the control of memory CD8+ T cells. AB - Previously we demonstrated that IL-15 and IL-2 control the number of memory CD8+ T cells in mice. IL-15 induces, and IL-2 suppresses the division of these cells. Here we show that CD25+CD4+ regulatory T cells play an important role in the IL-2 mediated control of memory phenotype CD8+ T cell number. In animals, the numbers of CD25+CD4+ T cells were inversely correlated with the numbers of memory phenotype CD8+ T cells with age. Treatment with anti-IL-2 caused CD25+CD4+ T cells to disappear and, concurrently, increased the numbers of memory phenotype CD8+ T cells. This increase in the numbers of CD8+ memory phenotype T cells was not manifest in animals lacking CD4+ cells. Importantly, adoptive transfer of CD25+CD4+ T cells significantly reduced division of memory phenotype CD8+ T cells. Thus, we conclude that CD25+CD4+ T cells are involved in the IL-2-mediated inhibition of memory CD8+ T cell division and that IL-2 controls memory phenotype CD8+ T cell numbers at least in part through maintenance of the CD25+CD4+ T cell population. PMID- 12084928 TI - Identification of gene function by cyclical packaging rescue of retroviral cDNA libraries. AB - Genes regulating responses in mammalian cells are often difficult to identify by functional cloning strategies limited to a single round of selection. Here we describe a strategy, cyclical packaging rescue (CPR), which allows rapid recovery and retransmission of retroviral cDNA libraries. CPR can be used not only with immortalized cell lines such as fibroblasts and Jurkat T cells, but also with primary B lymphocytes, which can be maintained only in short-term cultures. CPR allows for multiple rounds of selection and enrichment to identify cDNAs regulating responses in mammalian cells. Using CPR, five cDNAs were functionally cloned, which conferred protection against tumor necrosis factor alpha (TNFalpha) induced apoptosis in RelA(-/-) fibroblasts. Three of the genes, RelA, cellular FLICE-like inhibitory protein (c-FLIP), and a dominant-negative mutant of TNF receptor 1 arising through CPR afforded strong protection against apoptosis. Two of the genes identified, Dbs and Fas-associated death domain protein (FADD), previously identified as a proapoptotic molecule, afforded partial protection against TNFalpha-induced apoptosis. These results suggest that CPR is a versatile method that permits functional identification of both wild-type and dominant negative gene products that regulate cellular responses. PMID- 12084929 TI - Distinct recognition by two subsets of T cells of an MHC class II-peptide complex. AB - We examine here the nature of the differential recognition by CD4+ T cells of a single peptide from hen-egg white lysozyme (HEL) presented by I-A(k) class II MHC molecules. Two subsets of T cells (called A and B) interact with the same peptide, each in unique ways that reflect the nature of the complex of peptide and MHC. We show that the A and B set of T cells can be distinguished by their functional interaction with the three T cell receptor (TCR) contact residues of the bound peptide. The dominant peptide of HEL selected from processing is bound in a single register where a critical TCR contact residue is situated about the middle of the core segment of the peptide: all T cells establish functional contact with it. Three sets of T cells, however, can be distinguished by their differential recognition of two TCR contacts situated at the amino and carboxyl sides of the central TCR contact residue. Type A T cells, the conventional cells that see the peptide after processing of HEL, need to recognize all three TCR contact residues. In contrast, the type B T cells recognize the peptide given exogenously, but not when processed: these T cells recognize either one of the peripheral TCR contact residues, indicating a much more flexible interaction of peptide with I-A(k) molecules. We discuss the mode of generation of the various T cells and their biological relevance. PMID- 12084930 TI - The myelin basic protein gene is expressed in differentiated blood cell lineages and in hemopoietic progenitors. AB - Myelin basic proteins (MBP) are major constituents of the myelin sheath of oligodendrocytes and Schwann cells in the central nervous system and the peripheral nervous system, respectively. We previously showed that MBP-related transcripts are present in the bone marrow and the immune system. These mRNAs are transcribed from a region called 0', consisting of three exons, located upstream of the classical MBP exons; these three exons belong to the long MBP gene otherwise called "Golli-MBP." The most abundant of these mRNAs, now called HMBP (hemopoietic MBP), encompasses the sequence encoded by the region 0' plus exon 1 and part of intron 1 of the classic MBP gene. Antisera to recombinant HMBP proteins are immunoreactive with proteins of about 26-28 kDa in brain, thymus, and spleen. This report demonstrates that HMBP proteins are present in the vast majority (>95%) of thymic T cells, which express the corresponding transcripts, as do mature T cells from lymph nodes and spleen. HMBP mRNAs and proteins are also manifest in the majority of spleen B lymphocytes and in B cell lines. In addition to lymphoid cells, HMBP proteins are in all types of myeloid lineage cells, i.e., macrophages, dendritic cells, and granulocytes, as well as in megakaryocytes and erythroblasts. Finally, HMBP proteins are present in CD34+ bone marrow cells, and, furthermore, in highly proliferative cultures, these CD34+ cells express HMBP RNAs and proteins. Thus, MBP gene products are present both in the nervous system and in the entire hemopoietic system. PMID- 12084932 TI - Stable expression of calpain 3 from a muscle transgene in vivo: immature muscle in transgenic mice suggests a role for calpain 3 in muscle maturation. AB - Limb-girdle muscular dystrophy, type 2A (LGMD 2A), is an autosomal recessive disorder that causes late-onset muscle-wasting, and is due to mutations in the muscle-specific protease calpain 3 (C3). Although LGMD 2A would be a feasible candidate for gene therapy, the reported instability of C3 in vitro raised questions about the potential of obtaining a stable, high-level expression of C3 from a transgene in vivo. We have generated transgenic (Tg) mice with muscle specific overexpression of full-length C3 or C3 isoforms, which arise from alternative splicing, to test whether stable expression of C3 transgenes could occur in vivo. Unexpectedly, we found that full-length C3 can be overexpressed at high levels in vivo, without toxicity. In addition, we found that Tg expressing C3 lacking exon 6, an isoform expressed embryonically, have muscles that resemble regenerating or developing muscle. Tg expressing C3 lacking exon 15 shared this morphology in the soleus, but not other muscles. Assays of inflammation or muscle membrane damage indicated that the Tg muscles were not degenerative, suggesting that the immature muscle resulted from a developmental block rather than degeneration and regeneration. These studies show that C3 can be expressed stably in vivo from a transgene, and indicate that alternatively spliced C3 isoforms should not be used in gene-therapy applications because they impair proper muscle development. PMID- 12084931 TI - Urokinase-urokinase receptor interaction mediates an inhibitory signal for HIV-1 replication. AB - Elevated levels of soluble urokinase-type plasminogen activator (uPA) receptor, CD87/u-PAR, predict survival in individuals infected with HIV-1. Here, we report that pro-uPA (or uPA) inhibits HIV-1 expression in U937-derived chronically infected promonocytic U1 cells stimulated with phorbol 12-myristate 13-acetate (PMA) or tumor necrosis factor-alpha (TNF-alpha). However, pro-uPA did not inhibit PMA or TNF-alpha-dependent activation of nuclear factor-kB or activation protein-1 in U1 cells. Cell-associated HIV protein synthesis also was not decreased by pro-uPA, although the release of virion-associated reverse transcriptase activity was substantially inhibited, suggesting a functional analogy between pro-uPA and the antiviral effects of IFNs. Indeed, cell disruption reversed the inhibitory effect of pro-uPA on activated U1 cells, and ultrastructural analysis confirmed that virions were preferentially retained within cell vacuoles in pro-uPA treated cells. Neither expression of endogenous IFNs nor activation of the IFN-inducible Janus kinase/signal transducer and activator of transcription pathway were induced by pro-uPA. Pro-uPA also inhibited acute HIV replication in monocyte-derived macrophages and activated peripheral blood mononuclear cells, although with great inter-donor variability. However, pro-uPA inhibited HIV replication in acutely infected promonocytic U937 cells and in ex vivo cultures of lymphoid tissue infected in vitro. Because these effects occurred at concentrations substantially lower than those affecting thrombolysis, pro-uPA may represent a previously uncharacterized class of antiviral agents mimicking IFNs in their inhibitory effects on HIV expression and replication. PMID- 12084933 TI - Bucillamine, a thiol antioxidant, prevents transplantation-associated reperfusion injury. AB - Ischemia/reperfusion (I/R) injury is a serious potential threat to outcomes in organ transplantation and other clinical arenas in which there is temporary interruption of blood flow. I/R is a frequent cause of primary failure in organ transplantation. We hypothesized that the antioxidant bucillamine, a potent sulfhydryl donor, would protect against I/R injury in high-risk organ transplants. Because livers subjected to prolonged ischemia and very fatty livers are highly susceptible to severe I/R injury, we studied the effect of bucillamine in three animal models of liver transplantation: two ex vivo models of isolated perfused livers, either normal or fatty rat livers, and an in vivo model of syngenic orthotopic liver transplants in rats. In all models, livers were deprived of oxygen for 24 h before either ex vivo reperfusion or transplantation. In the ex vivo models, bucillamine treatment significantly improved portal vein blood flow and bile production, preserved normal liver architecture, and significantly reduced liver enzyme release and indices of oxidative stress. Moreover, bucillamine treatment significantly increased levels of reduced glutathione in the liver and lowered levels of oxidized glutathione in both liver and blood. In rats subjected to liver transplants, bucillamine significantly enhanced survival and protected against hepatic injury. Possible mechanisms of this protection include prevention of excessive accumulation of toxic oxygen species, interruption of redox signaling in hepatocytes, and inhibition of macrophage activation. This study demonstrates the potential utility of bucillamine or other cysteine-derived thiol donors for improving outcomes in organ transplantation and other clinical settings involving I/R injury. PMID- 12084934 TI - Isolated allogeneic bone marrow-derived mesenchymal cells engraft and stimulate growth in children with osteogenesis imperfecta: Implications for cell therapy of bone. AB - Treatment with isolated allogeneic mesenchymal cells has the potential to enhance the therapeutic effects of conventional bone marrow transplantation in patients with genetic disorders affecting mesenchymal tissues, including bone, cartilage, and muscle. To demonstrate the feasibility of mesenchymal cell therapy and to gain insight into the transplant biology of these cells, we used gene-marked, donor marrow-derived mesenchymal cells to treat six children who had undergone standard bone marrow transplantation for severe osteogenesis imperfecta. Each child received two infusions of the allogeneic cells. Five of six patients showed engraftment in one or more sites, including bone, skin, and marrow stroma, and had an acceleration of growth velocity during the first 6 mo postinfusion. This improvement ranged from 60% to 94% (median, 70%) of the predicted median values for age- and sex-matched unaffected children, compared with 0% to 40% (median, 20%) over the 6 mo immediately preceding the infusions. There was no clinically significant toxicity except for an urticarial rash in one patient just after the second infusion. Failure to detect engraftment of cells expressing the neomycin phosphotransferase marker gene suggested the potential for immune attack against therapeutic cells expressing a foreign protein. Thus, allogeneic mesenchymal cells offer feasible posttransplantation therapy for osteogenesis imperfecta and likely other disorders originating in mesenchymal precursors. PMID- 12084935 TI - Human alpha-synuclein-harboring familial Parkinson's disease-linked Ala-53 --> Thr mutation causes neurodegenerative disease with alpha-synuclein aggregation in transgenic mice. AB - Mutations in alpha-synuclein (alpha-Syn) cause Parkinson's disease (PD) in a small number of pedigrees with familial PD. Moreover, alpha-Syn accumulates as a major component of Lewy bodies and Lewy neurites, intraneuronal inclusions that are neuropathological hallmarks of PD. To better understand the pathogenic relationship between alterations in the biology of alpha-Syn and PD-associated neurodegeneration, we generated multiple lines of transgenic mice expressing high levels of either wild-type or familial PD-linked Ala-30 --> Pro (A30P) or Ala-53 -> Thr (A53T) human alpha-Syns. The mice expressing the A53T human alpha-Syn, but not wild-type or the A30P variants, develop adult-onset neurodegenerative disease with a progressive motoric dysfunction leading to death. Pathologically, affected mice exhibit neuronal abnormalities (in perikarya and neurites) including pathological accumulations of alpha-Syn and ubiquitin. Consistent with abnormal neuronal accumulation of alpha-Syn, brain regions with pathology exhibit increases in detergent-insoluble alpha-Syn and alpha-Syn aggregates. Our results demonstrate that the A53T mutant alpha-Syn causes significantly greater in vivo neurotoxicity as compared with other alpha-Syn variants. Further, alpha-Syn dependent neurodegeneration is associated with abnormal accumulation of detergent insoluble alpha-Syn. PMID- 12084936 TI - Trace fear conditioning involves hippocampal alpha5 GABA(A) receptors. AB - The heterogeneity of gamma-aminobutyric acid type A (GABA(A)) receptors contributes to the diversity of neuronal inhibition in the regulation of information processing. Although most GABA(A) receptors are located synaptically, the small population of alpha5GABA(A) receptors is largely expressed extrasynaptically. To clarify the role of the alpha5GABA(A) receptors in the control of behavior, a histidine-to-arginine point mutation was introduced in position 105 of the murine alpha5 subunit gene, which rendered the alpha5GABA(A) receptors diazepam-insensitive. Apart from an incomplete muscle relaxing effect, neither the sedative, anticonvulsant, nor anxiolytic-like activity of diazepam was impaired in alpha5(H105R) mice. However, in hippocampal pyramidal cells, the point mutation resulted in a selective reduction of alpha5GABA(A) receptors, which altered the drug-independent behavior. In line with the role of the hippocampus in certain forms of associative learning, trace fear conditioning, but not delay conditioning or contextual conditioning, was facilitated in the mutant mice. Trace fear conditioning differs from delay conditioning in that the conditioned and unconditioned stimulus are separated by a time interval. Thus, the largely extrasynaptic alpha5GABA(A) receptors in hippocampal pyramidal cells are implicated as control elements of the temporal association of threat cues in trace fear conditioning. PMID- 12084937 TI - Altered accumbens neural response to prediction of reward associated with place in dopamine D2 receptor knockout mice. AB - Midbrain dopaminergic activity seems to be important in forming the prediction of future events such as rewards. The nucleus accumbens (NAc) plays an important role in the integration of reward with motor function, and it receives dense dopamine innervation and extensive limbic and cortical afferents. Here, we examined the specific role of the dopamine D2 receptor (D2R) in mediating associative learning, locomotor activity, and regulating NAc neural responses by using D2R-knockout (KO) mice and their wild-type littermates. D2R-KO mice displayed reduced locomotor activity and slower acquisition of a place-learning task. D2R-KO eliminated the prereward inhibitory response of neurons in the NAc. In contrast, an increased number of neurons in D2R-KO mice displayed place related activity. These results provide evidence that D2R in the NAc participates in coding for a specific type of neural response to incentive contingencies and partly in spatial learning. PMID- 12084938 TI - Deciphering peripheral nerve myelination by using Schwann cell expression profiling. AB - Although mutations in multiple genes are associated with inherited demyelinating neuropathies, the molecular components and pathways crucial for myelination remain largely unknown. To approach this question, we performed genome-wide expression analysis in several paradigms where the status of peripheral nerve myelination is dynamically changing. Anchor gene correlation analysis, a form of microarray analysis that integrates functional information, using correlation based clustering, with a statistically rigorous test, the Westfall and Young step down algorithm, was applied to this data set. Biological pathways active in myelination, genes encoding proteins involved in myelin synthesis, and genes whose mutation results in myelination defects were identified. Many known genes and previously uncharacterized ESTs not heretofore associated with myelination were also identified. One of these ESTs, MASR (myelin-associated SUR4 protein), encodes a member of the SUR4 family of fatty acid desaturases, enzymes involved in elongation of very long chain fatty acids. Its specific localization in myelinating Schwann cells indicates a crucial role for MASR in normal myelin lipid synthesis. PMID- 12084939 TI - Leukemia inhibitory factor inhibits neuronal terminal differentiation through STAT3 activation. AB - The discovery of stem cells in the adult central nervous system raises questions concerning the neurotrophic factors that regulate postnatal neuronal development. Olfactory receptor neurons (ORNs) are a useful model, because they are capable of robust neurogenesis throughout adulthood. We have investigated the role of leukemia inhibitory factor (LIF) in postnatal neuronal development by using ORNs as a model. LIF is a multifunctional cytokine implicated in various aspects of neuronal development, including phenotype determination, survival, and in response to nerve injury. LIF-deficient mice display significant increases, both in the absolute amount and in the number of cells expressing olfactory marker protein, a marker of mature ORNs. The maturation of ORNs was significantly inhibited by LIF in vitro. LIF activated the STAT3 pathway in ORNs, and transfection of ORNs with a dominant negative form of STAT3 abolished the effect of LIF. These findings demonstrate that LIF negatively regulates ORN maturation via the STAT3 pathway. Thus, LIF plays a critical role in controlling the transition of ORNs to maturity. Consequently, a population of ORNs is maintained in an immature state to facilitate the rapid repopulation of the olfactory epithelium with mature neurons during normal cell turnover or after injury. PMID- 12084940 TI - Cocaine sensitization and reward are under the influence of circadian genes and rhythm. AB - Investigations using the fruit fly Drosophila melanogaster have shown that the circadian clock gene period (Per) can influence behavioral responses to cocaine. Here we show that the mouse homologues of the Drosophila Per gene, mPer1 and mPer2, modulate cocaine sensitization and reward, two phenomena extensively studied in humans and animals because of their importance for drug abuse. In response to an acute cocaine injection mPer1 and mPer2 mutant mice as well as wild-type mice exhibited an approximately 5-fold increase in activity compared with saline control levels, showing that there is no initial difference in sensitivity to acute cocaine administration in Per mutants. After repeated cocaine injections wild-type mice exhibited a sensitized behavioral response that was absent in mPer1 knockout mice. In contrast, mPer2 mutant mice exhibited a hypersensitized response to cocaine. Conditioned place preference experiments revealed similar behavioral reactions: mPer1 knockout mice showed a complete lack of cocaine reward whereas mPer2 mutants showed a strong cocaine-induced place preference. In another set of experiments, we tested C57/BL6J mice at different Zeitgeber times and found that cocaine-induced behavioral sensitization and place preference are under the control of the circadian clock. In conclusion, we demonstrate that processes involved in cocaine addiction depend on the circadian rhythm and are modulated in an opposing manner by mPer1 and mPer2 genes. PMID- 12084941 TI - Inosine induces axonal rewiring and improves behavioral outcome after stroke. AB - Cerebral infarct (stroke) often causes devastating and irreversible losses of function, in part because of the brain's limited capacity for anatomical reorganization. The purine nucleoside inosine has previously been shown to induce neurons to express a set of growth-associated proteins and to extend axons in culture and in vivo. We show here that in adult rats with unilateral cortical infarcts, inosine stimulated neurons on the undamaged side of the brain to extend new projections to denervated areas of the midbrain and spinal cord. This growth was paralleled by improved performance on several behavioral measures. PMID- 12084943 TI - Suppression and acceleration of cell elongation by integration of xyloglucans in pea stem segments. AB - Xyloglucan is a key polymer in the walls of growing plant cells. Using split pea stem segments and stem segments from which the epidermis had been peeled off, we demonstrate that the integration of xyloglucan mediated by the action of wall bound xyloglucan endotransglycosylase suppressed cell elongation, whereas that of its fragment oligosaccharide accelerated it. Whole xyloglucan was incorporated into the cell wall and induced the rearrangement of cortical microtubules from transverse to longitudinal; in contrast, the oligosaccharide solubilized xyloglucan from the cell wall and maintained the microtubules in a transverse orientation. This paper proposes that xyloglucan metabolism controls the elongation of plant cells. PMID- 12084944 TI - Disease surveillance in recombining pathogens: multilocus genotypes identify sources of human Coccidioides infections. AB - Molecular surveillance of pathogenic microbes works by genotyping isolates with DNA fingerprinting techniques and then using these genotypes to assign individuals to populations. Clonality is assumed in many fingerprinting studies, although this assumption has been shown to be false for many organisms. To accommodate recombining organisms into surveillance programs, methods using population allele frequencies in combination with individual multilocus genotypes are necessary. Here, we develop a statistical method appropriate for haploid recombining microbes that allows individuals to be assigned to populations. We illustrate the usefulness of this technique by inferring the source populations for Coccidioides isolates recovered from patients treated outside the endemic area of Coccidioides sp., the etiological agents of human coccidioidomycosis, but with a travel history including visits to one or more endemic areas. PMID- 12084942 TI - Two immediate-early pathogen-responsive members of the AtCMPG gene family in Arabidopsis thaliana and the W-box-containing elicitor-response element of AtCMPG1. AB - The Arabidopsis thaliana genome contains at least 50 predicted AtCMPG genes. The encoded protein family is defined by a common domain possessing four strictly conserved amino acid residues [Cys, Met, Pro, and Gly (CMPG)] that designate the family. Two members, AtCMPG1 and AtCMPG2, with high sequence similarity to the previously described, immediate-early pathogen-responsive PcCMPG1 gene from Petroselinum crispum were selected for analysis of their expression modes and defense-related promoter elements. Among the most striking similarities with PcCMPG1 were immediate-early transcriptional activation on infection or treatment with a pathogen-derived elicitor and the functional importance of a W-box containing AtCMPG1 promoter element. Remarkably, this strongly pathogen/elicitor responsive element, F, did not respond to wounding, in contrast to the AtCMPG1 promoter itself. Comparative analysis, both within the A. thaliana genome and across species, provided further insight into the large structural diversity of W box-containing elements. Possible roles of AtCMPG proteins in regulatory processes are discussed with reference to a large variety of family members, partly with assigned functions, from plants as well as animals. PMID- 12084945 TI - Improving the quality of care for children. PMID- 12084947 TI - Exploring the power of Powerpoint. PMID- 12084946 TI - Respiratory distress in neonates with special reference to pneumonia. AB - OBJECTIVE: (i) To find causes of respiratory distress in neonates brought to a referral neonatal unit with symptoms suggestive of respiratory disorder; (ii) to evaluate clinical signs for diagnosis of neonatal pneumonia; (iii) To determine bacterial etiology of neonatal pneumonia; and (iv) To determine indicators of fatality in neonatal pneumonia. DESIGN: Prospective descriptive. SETTING: Referral neonatal unit of a teaching hospital. SUBJECTS: 150 neonates admitted with respiratory symptoms consecutively. METHODS: All neonates presenting with respiratory symptoms were included in the study. The diagnosis of the cause of respiratory distress was based on guidelines recommended by the National Neonatology Forum. Clinical features, FiO2 requirement, sepsis screen, X-ray chest, blood culture and antibiotic sensitivity, arterial blood gases and other relevant investigations were documented in a structured proforma. The neonates were regularly followed up for outcome. Multivariate unweighted logistic regression was done to find out the indicators of fatality in neonatal pneumonia for those variables which were significantly associated with outcome on univariate analysis. RESULTS: Pneumonia was found to be the most common cause (68.6%) of respiratory distress in neonates. Other conditions included hydline membrane disease (HMD), transient tachypneia of new born (TTNB), birth asphyxia with hypoxic ischemic encephalopathy (HIE) and meconium aspiration syndrome (MAS). Clinical signs and symptoms were non specific and did not differentiate between pneumonia and other causes of respiratory distress. Respiratory rate was less than 60 per minute in 11.6% neonates with pneumonia. The most common organism responsible for neonatal pneumonia was Klebsiella pneumoniae. Chest X ray was clear in 15% of neonates with pneumonia. On univariate analysis weight < 2000 g, gestation age < 34 weeks, age at presentation < 72 hours, lethargy, absent neonatal reflexes, shock, positive C-reactive protein, positive ventilatory support, blood culture positivity, Silverman Score >3, FiO2 >40%, pH < 7.2, alveolar arterial gradient (AaDO2) > 250 mmHg and arterial alveolar tension ratio (a/A) of < 0.25 were significantly associated with mortality in neonates with pneumonia. However, on multivariate analysis, only AaDO2 of > 250 mmHg was found to be independent predictor of fatality in neonatal pneumonia. CONCLUSION: Pneumonia was the most common cause of respiratory distress in neonates. Clinical features and X-ray chest missed the diagnosis of pneumonia in 15 cases and had to be corroborated with sepsis screen and blood culture. AaDO2 > 250 mmHg was an independent predictor of fatality in neonatal pneumonia PMID- 12084948 TI - Anthracycline associated cardiac toxicity in children with malignancies. PMID- 12084950 TI - Human cytomegalovirus perinatal infections in a tertiary care setting. PMID- 12084949 TI - Measles vaccine efficacy evaluated by case reference technique. PMID- 12084951 TI - Efficacy of alpha, beta - Arteether in children with cerebral malaria in forested tribal belt. PMID- 12084952 TI - Prevalence of positive throat cultures for group A beta-hemolytic streptococci among school children in Istanbul. PMID- 12084953 TI - Congenital hemolytic anemia in Bangladesh: types and clinical manifestations. PMID- 12084954 TI - Perforated duodenal ulcer in a child. PMID- 12084955 TI - Succimer in symptomatic lead poisoning. PMID- 12084956 TI - Resistant malaria in a neonate. PMID- 12084957 TI - Biliary peritonitis: a rare presentation of perforated choledochal cyst. PMID- 12084958 TI - Ash leaf macules. PMID- 12084959 TI - Mucopolysaccharidosis Types IV and VI. PMID- 12084960 TI - HIV wasting syndrome. PMID- 12084961 TI - Podophyllin poisoning. PMID- 12084962 TI - Hypernatremic dehydration in a neonate. PMID- 12084964 TI - An indigenous leucocyte esterase test along with Pandy's test for the diagnosis of bacterial meningitis. PMID- 12084966 TI - Cucumber: a model angiosperm for mitochondrial transformation? AB - Plants possess three major genomes, carried in the chloroplast, mitochondrion, and nucleus. The chloroplast genomes of higher plants tend to be of similar sizes and structure. In contrast both the nuclear and mitochondrial genomes show great size differences, even among closely related species. The largest plant mitochondrial genomes exist in the genus Cucumis at 1500 to 2300 kilobases, over 100 times the sizes of the yeast or human mitochondrial genomes. Biochemical and molecular analyses have established that the huge Cucumis mitochondrial genomes are due to extensive duplication of short repetitive DNA motifs. The organellar genomes of almost all organisms are maternally transmitted and few methods exist to manipulate these important genomes. Although chloroplast transformation has been achieved, no routine method exists to transform the mitochondrial genome of higher plants. A mitochondrial-transformation system for a higher plant would allow geneticists to use reverse genetics to study mitochondrial gene expression and to establish the efficacy of engineered mitochondrial genes for the genetic improvement of the mitochondrial genome. Cucumber possesses three unique attributes that make it a potential model system for mitochondrial transformation of a higher plant. Firstly, its mitochondria show paternal transmission. Secondly, microspores possess relatively few, huge mitochondria. Finally, there exists in cucumber unique mitochondrial mutations conditioning strongly mosaic (msc) phenotypes. The msc phenotypes appear after regeneration of plants from cell culture and sort with specific rearranged and deleted regions in the mitochondrial genome. These mitochondrial deletions may be a useful genetic tool to develop selectable markers for mitochondrial transformation of higher plants. PMID- 12084967 TI - Cytogenetic analysis of an asymmetric potato hybrid. AB - An asymmetric potato hybrid and its parental lines were cytogenetically examined. DAPI (4'-6-diamidino-2-phenylindole) staining was used to count chromosomes in all analysed lines and revealed the presence of minichromosomes in the hybrid genome. Fluorescent in situ hybridization (FISH) with rDNA sequence as a probe helped to determine the ploidy level of analysed lines and revealed that none of the minichromosomes contains rDNA repeats. CMA (chromomycin A3) band occurred to be a new chromosome marker that identifies potato chromosome No.1. It was possible to detect a deletion in one of four chromosomes No. 1 of the asymmetric potato hybrid. On the basis of these analyses a karyotype of the asymmetric hybrid was constructed. PMID- 12084968 TI - Interaction of gene effects with environments for malting quality of barley doubled haploids. AB - Barley doubled haploids covering a wide range of malting quality, along with their parental cultivars and F2, F3 hybrids, were investigated in six environments (three locations, two years) to study the genotype-environment (G x E) interaction structure and the influence of environments on additive, dominance and epistatic gene effects. Grain and malt characters, such as 1000-grain weight, percentage of plump kernels, malt extract yield, protein content, Kolbach index and malt fine-coarse difference (FCD), were measured. Main effects for genetic parameters were estimated and regression analysis was used to explain the interaction of gene effects with environments. The results show that additive effects had the greatest interaction with environments for all the analysed traits, but only for malt characters this interaction was linear. Interaction of dominance effects was much lower and only in the case of 1000-grain weight, protein content and Kolbach index it proved to be significant. The results suggest that effects of heterozygous loci are more stable in contrasting environments than effects of homozygous loci. PMID- 12084969 TI - Genetic relationship between Polish and Chinese strains of the mushroom Agaricus bisporus (Lange) Sing., determined by the RAPD method. AB - The genetic relationship between twenty-six strains of Agaricus bisporus were analysed by the RAPD (random amplified polymorphic DNA) method. DNA amplification was performed with the use of twelve arbitrary 10-mer primers. Four primers, which gave polymorphic band patterns were chosen for RAPD analysis. In total, they gave 24 distinguishable bands, of which nine were polymorphic. The conducted research showed that there is a great genetic similarity among the examined strains. Low polymorphism of the strains may be a proof of a limited genetic pool used in the cultivation of those strains. PMID- 12084970 TI - Morphological diversity of androgenic carrot plants. AB - The aim of this study was to evaluate phenotypic variation of R0 androgenic plants obtained from four seed sources and donor plants by anther culture. Several morphological traits (leaf size, petiole length, leaf division, cortex colour) and the range of diversity were evaluated. There was large variation in all traits among the donor varieties. Especially leaf division and cortex colour differed significantly among the androgenic plants that came from different seed sources. The plants regenerated from four donor plants of variety 62 were significantly different in most traits except for leaf width and cortex colour. Evaluation of R1 plants will demonstrate whether the R0 variation observed is due to genetic variation or physiological differences from tissue culture. PMID- 12084971 TI - Genetic aspects of twinning in cattle. AB - Twinning in cattle ranges from about 1% for beef breeds to about 4% for dairy breeds. The incidence of double births may have both positive and negative effects, which mainly depends on the purpose for which the cattle are raised. Because of freemartinism, as well as management problems connected e.g. with a greater risk of dystocia and retained placenta, it is an undesirable trait in dairy herds. In beef cattle, however, twinning can considerably increase the efficiency of production. Low heritability, a long generation interval for progeny testing, sex-limited expression and an unfavourable correlation with milk yield make twinning difficult to control by selection. Hence, it is the type of trait for which the identification of the genetic marker - quantitative trait loci (QTL) linkage and the implementation of marker-assisted selection in breeding strategies are expected to be especially beneficial. Searching for QTL influencing the reproductive rate in cattle was performed mainly in the US Meat Animal Research Center twinning herd and in the commercial Norwegian cattle population. Among several genome regions that appear to control twinning and ovulation rates, the most interesting seem to be chromosomes 5, 7, 19 and 23. PMID- 12084972 TI - Application of a mixed inheritance model to the detection of quantitative trait loci in swine. AB - The primary goal of this study was to investigate statistical properties of a mixed inheritance model for the localization of quantitative trait loci (QTL). This is based on the analysis of phenotypic data for the amount of intramuscular fat (IMF) scored on 305 individuals originating from a cross between Duroc and Norwegian Landrace breeds. Marker genotype information is available for F1 and F2 generations. Statistical procedures compared involve i) the interval mapping, ii) the composite interval mapping, iii) a regression method, and iv) a mixed inheritance model accounting for a random animal additive genetic effect and relationships between individuals. The basic statistical properties of the latter approach are then assessed using Monte Carlo simulations showing slight unconservativeness as compared to chi(2)2df and reasonable power to detect QTL of moderate effects. In the analysis of IMF data, the significant evidence for the existing QTL is detected on chromosome 6. A chromosomal region recommended for a second-step fine mapping analysis is identified between markers SW1823 and S0228, based on three types of confidence intervals derived by using: i) the Jackknife algorithm, ii) the numerical variance approximation, and iii) the LOD score approach. The Jackknife algorithm was additionally used to quantify each family's contribution to the test statistic and to the estimate of QTL position. PMID- 12084973 TI - Analysis of associations of PIT1 genotypes with growth, meat quality and carcass composition traits in pigs. AB - PIT1 was chosen as a candidate gene to investigate its associations with growth, meat quality and carcass composition traits in the pig. PIT1 is known as the pituitary-specific activator of the growth hormone in several mammals. Furthermore, PIT1 is a positive regulatory factor of prolactin and thyroid stimulating hormone beta. PIT1 is a member of the POU-domain family of genes and is located on porcine chromosome 13. Two informative three-generation families of the University of Hohenheim were used for the presented investigations. The families were based on crosses of the European Wild boar (W) x Pietrain (P) and Meishan (M) x Pietrain (P). Each family included 310 F(2) animals. A RsaI (PCR) RFLP described by Yu et al. (1994) was used for genotyping the animals. Altogether over 50 parameters of growth, meat quality, carcass composition and stress susceptibility were evaluated concerning their associations with RsaI PCR RFLP. The statistical model of association analyses was used including fixed effects of sex, family, PIT1 genotypes and covariate age at slaughter. Taking the significance level of p < 0.05 as the basis, fourteen traits of growth and carcass composition were associated with PIT1 genotypes in family W x P. Results from this study suggest that there are contributions of PIT1 gene to variations in the analysed performance traits in pigs. The influence of PIT1 genotypes could not be confirmed under the supposition of a genome-wide test limit. PMID- 12084974 TI - Distribution of polymorphic forms at the porcine GH locus in a population of day 10 pig embryos. AB - The present study describes an analysis of genotype and allele distribution at the porcine GH locus among day-10 pig embryos. Embryos were collected post mortem from 6 crossbred (Danish Landrace x Yorkshire) sows inseminated with mixed Duroc semen and individually frozen for later analysis. After extraction, DNA was subjected to PCR amplification and restriction analysis with Msp I and Hae II enzymes. The genotype frequencies were: Msp I CD 0.17, DD 0.83; and Hae II AA 0.33, AB 0.58; and BB 0.09. The Msp I CC genotype was not found among analysed embryos. To our knowledge, this is the first report on the genotype and allele distribution at the GH locus among early pig embryos. PMID- 12084975 TI - Recent advances in understanding of the molecular basis of anhidrotic ectodermal dysplasia: discovery of a ligand, ectodysplasin A and its two receptors. AB - Recent developments of the investigations on the molecular basis of anhidrotic ectodermal dysplasia are reviewed. Identification of the major product of the EDA gene (ectodysplasin A), a protein belonging to a group of TNF ligands, and molecular cloning of the cDNA, encoding its receptor (EDAR), a member of the TNF receptor family, are presented. The role of an alternative EDA receptor, localised on the X chromosome (XEDAR) in the developmental control of the differentiation of skin appendages, is discussed. Recent findings have elucidated the cause of the autosomal forms of EDA, both dominant and recessive, and indicated an important role of a signal transduction pathway involving a protein product of the NEMO gene and the transcription factor NFkappaB in the differentiation of skin appendages. PMID- 12084976 TI - Transition C2718T in the AR gene, resulting in generation of a termination codon and truncated form of the androgen receptor, causes complete androgen insensitivity syndrome. AB - The action of testosterone and 5 alpha-dihydrotestosterone are essential to the development of the male phenotype. Patients with karyotype 46,XY, resistant to these hormones, exhibit a wide spectrum of phenotypes: from phenotypic female, through a range of incomplete masculinization, to under-virilized, infertile man. These disturbances are caused by mutations in the androgen receptor gene (AR). We studied a 46,XY fenotypic female with typical symptoms of Complete Androgen Insensitivity Syndrome (CAIS). Multiple temperature single-stranded conformation polymorphism (MSSCP) and sequence analysis of exon 6 of the AR gene in a patient revealed a C2718T transition causing R786X mutation in the loop between helices VII and VIII of the LBD of the androgen receptor. The R786X mutation has been described in a patient with CAIS only once and no such mutations have been described in Eastern Europe. PMID- 12084977 TI - Trisomy 8 mosaicism syndrome. AB - Authors present the case of a 15-year-old boy assessed for Marfan syndrome for many years. The child was treated because of skeletal defects, mild mental deficiency and dysmorphic features of face. Chromosomal analysis showed a trisomy 8 mosaicism. PMID- 12084978 TI - Cell density-dependent changes of glycosphingolipid biosynthesis in cultured human skin fibroblasts. AB - In this study, the glycosphingolipid biosynthesis was investigated in the sparse and the confluent cell populations of cultured human skin fibroblasts. The human skin fibroblast cell populations were metabolically pulse labeled with (14)C galactose (48 h). The amounts of (14)C-radioactivity (cpm) incorporated into extracted and purified total cellular glycosphingolipid fractions were counted by beta-scintillation and the individual glycosphingolipid species were separated by high performance thin layer chromatography and visualized by autoradiography. The relative labeling (%) of individual newly synthesized glycosphingolipid species was detected by densitometric scanning of autoradiographic glycosphingolipid patterns. The incorporation of (14)C-label into total glycosphingolipids per cell increased significantly as the cell-density increased, referring to five fold higher rate of glycosphingolipid biosynthesis de novo in cells at confluency vs. sparse populations. The total newly synthesized glycosphingolipid pattern (100%) of sparse cell populations showed a significant predominance of the gangliosides (70%) over the neutral glycosphingolipids (30%), with ganglioside GM2 as the major species followed by monohexosyl-ceramide. Oppositely, the newly synthesized neutral glycosphingolipids (67%) predominated over the gangliosides (33%) in cells at confluency (contact inhibition). Cells reaching confluency were characterized by: (a) a dramatic increase of absolute amount of all newly synthesized neutral glycosphingolipid species, particularly the most abundant monohexosyl-ceramide and trihexosyl-ceramide, but also of the ganglioside GM3; (b) a drastic decrease of absolute amount of newly synthesized ganglioside GM2. The specific shift in newly synthesized glycosphingolipid pattern in cells reaching confluency suggests a down-regulation of biosynthetic pathway primarily at the level of N-acetylgalactosaminyl-transferase. A possible involvement of glycosphingolipids in cell density-dependent regulation of cell growth through establishment of the direct intermolecular intermembrane interactions is discussed. PMID- 12084979 TI - Identification of a cDNA encoding a plant Lewis-type alpha1,4-fucosyltransferase. AB - Recently, it has been found that plants, including tomato (Lycopersicon esculentum), express the Lewis-a epitope, Galbeta1,3(Fucalpha1,4)GlcNAc, on some N-glycans. By searching the EST database, it was possible to identify a tomato cDNA encoding a protein, designated FucTC, of 413 amino acids with homology to plant and mammalian alpha1,3/4-fucosyltransferases. The cDNA was expressed in Pichia pastoris and the recombinant enzyme was found to transfer fucose from GDP Fuc (K(m) 16 microM) to lacto-N-tetraose (Galbeta1,3GlcNAcbeta1,3Galbeta1,4Glc; K(m) 80 microM) as well as to beta1,3- and beta1,4-galactosylated N-glycans. It is concluded that FucTC is responsible for the biosynthesis of Lewis-a on N glycans in tomato. PMID- 12084981 TI - Identification by saturation mutagenesis of a single residue involved in the alpha-galactosidase AgaB regioselectivity. AB - The alpha-galactosidase AgaB of Bacillus stearothermophilus displays a major 1,6 and a minor 1,3 regioselectivity. The wild-type enzyme was subjected to directed evolution (random mutagenesis and in vitro recombination) using a double screening strategy based on the elimination of the 1,6 regioselectivity and the analysis by TLC of the transglycosylation products. One of the AgaB mutants (E500) exhibited a new 1,2 regioselectivity and a rather high level of transglycosylation. The corresponding gene contains 10 mutations compared to the agaB gene and we demonstrated by saturation mutagenesis that the G442R substitution strongly contributes to the emergence of this new regioselectivity. Moreover, other single point mutations at this position led to new mutants displaying other kinds of regioselectivity demonstrating the importance of this position in the subtle kinetic control of transglycosylation. PMID- 12084980 TI - Effect of carbohydrate structure on biological activity of artificially N glycosylated eel calcitonin. AB - To reveal the function of the carbohydrate portion of glycopeptides and glycoproteins, we chemo-enzymatically synthesized artificially N-glycosylated derivatives of eel calcitonin and studied their three-dimensional structure and biological activity. The CD and NMR spectra in trifluoroethanol-H(2)O solution showed that the glycosylation did not change the three-dimensional structure. All the derivatives retained the strong in vivo hypocalcemic activity of calcitonin. However, the relative activity was dependent on the structure of the attached carbohydrate. The single GlcNAc attachment best enhanced the activity, while larger carbohydrates decreased the activity. This relative activity order of compounds could be partly explained by their calcitonin-receptor binding affinity, though the affinity of the GlcNAc derivative did not exceed that of calcitonin. The enhanced hypocalcemic activity of the GlcNAc derivative was explained by its altered biodistribution. The GlcNAc attachment caused calcitonin to escape from the trap at the liver during the early circulation. Thus, the glycosylation was shown to modulate the biological activity of calcitonin depending on the carbohydrate structure without a change in the peptide backbone conformation. PMID- 12084982 TI - In vivo fucosylation of lacto-N-neotetraose and lacto-N-neohexaose by heterologous expression of Helicobacter pylori alpha-1,3 fucosyltransferase in engineered Escherichia coli. AB - We report here the in vivo production of type 2 fucosylated-N-acetyllactosamine oligosaccharides in Escherichia coli. Lacto-N-neofucopentaose Galbeta1 4GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3)Glc, lacto-N-neodifucohexaose Galbeta1 4(Fucalpha1-3)Glc-NAcbeta1-3Galbeta1-4(Fucalpha1-3)Glc, and lacto-N neodifucooctaose Galbeta1-4GlcNAcbeta1-3Galbeta1-4(Fucalpha1-3)GlcNAcbeta1 3Galbeta1-4(Fucalpha1-3)Glc were produced from lactose added in the culture medium. Two of them carry the Lewis X human antigen. High cell density cultivation allowed obtaining several grams of fucosylated oligosaccharides per liter of culture. The fucosylation reaction was catalyzed by an alpha-1,3 fucosyltransferase of Helicobacter pylori overexpressed in E. coli with the genes lgtAB of N. meningitidis. The strain was genetically engineered in order to provide GDP-fucose to the system, by genomic inactivation of gene wcaJ involved in colanic acid synthesis and overexpression of RcsA, positive regulator of the colanic acid operon. To prevent fucosylation at the glucosyl residue, lactulose Galbeta1-4Fru was assayed in replacement of lactose. Lactulose-derived oligosaccharides carrying fucose were synthesized and characterized. Fucosylation of the fructosyl residue was observed, indicating a poor acceptor specificity of the fucosyltransferase of H. pylori. PMID- 12084983 TI - Monoclonal antibody (5F3) defining renal cell carcinoma-associated antigen disialosyl globopentaosylceramide (V3NeuAcIV6NeuAcGb5), and distribution pattern of the antigen in tumor and normal tissues. AB - Renal cell carcinoma (RCC) has been characterized by high expression of three types of disialogangliosides: two based on lacto-series type 1 structure (disialosyl Lc(4), GalNAc disialosyl Lc(4)), the other based on globo-series structure (disialosyl globopentaosylceramide; disialosyl Gb5). The present study established a mAb, 5F3, directed to disialosyl Gb5. 5F3 was established after immunization with RCC cell line ACHN. The major disialoganglioside antigen isolated from ACHN cells, showing specific reactivity with 5F3, was characterized unequivocally as disialosyl Gb5 (V(3)NeuAcIV(6)NeuAcGb5) by identification of the core structure as globopentaosylceramide (Gb5) after enzymatic and acid hydrolysis, and by 2-dimensional (1)H-NMR spectroscopy. 5F3 does not react with monosialosyl Gb5 (V(3)NeuAcGb5), Gb5, or any lacto-series structures. 5F3 strongly stained 19 of 41 cases of primary RCC tissue. It reacted with proximal tubules (but not distal tubules) of kidney, microglial cells of cerebrum and cerebellum, goblet cells of stomach and intestine, smooth muscle of various organs. It did not react with parenchymatous cells of various organs, except for kidney epithelia and prostate stroma. Immunostaining of RCC tissue by mAb 5F3, in combination with staining by other antibodies directed to globo-series and lacto series structures, has prognostic significance in defining metastatic potential of RCC. PMID- 12084984 TI - Influence of TNFalpha on the sialylation of mucins produced by a transformed cell line MM-39 derived from human tracheal gland cells. AB - In order to investigate the influence of inflammation on the peripheral glycosylation of airway mucins, a human respiratory glandular cell line (MM-39) was treated by TNFalpha. The expression and the activity of sialyl- and fucosyl transferases, involved in the biosynthesis of peripheral carbohydrate determinants like sialyl-Lewis x, were investigated by RT-PCR and by HPAEC respectively. The mRNA steady-state level of sialyl- (ST3Gal III) and of fucosyl- (FUT3) transferases was moderately up-regulated by TNFalpha; a 52% increase of alpha2,3-sialyltransferase activity was also observed in TNFalpha-stimulated MM 39 cells. After metabolic radio-labelling with [(3)H]glucosamine and [(3)H]fucose, the mucins released in the culture supernatant were purified by Sepharose CL-4B, density-gradient centrifugation and treatment with glycosaminoglycans-degrading enzymes. The mucins, released in the culture supernatant from control MM-39 cells, were constituted by two populations of molecules having the same 1.39-1.44 mg/ml density but carrying either high or low amounts of sialic acid residues at their periphery. TNFalpha was able to increase the sialylation of the weakly sialylated mucins. This effect and the enhancement of the alpha2,3-sialyltransferase activity by TNFalpha argue in favour of a regulation of the mucin sialylation by this pro-inflammatory cytokine. Despite the moderate overexpression of FUT3, no fucosylation of mucins produced by MM-39 cells was induced by TNFalpha. In conclusion, the influence of TNFalpha on the sialylation of mucins could explain why the mucins from infected patients suffering either from cystic fibrosis or from chronic bronchitis are more sialylated. PMID- 12084986 TI - [New diagnostic frontiers in Brucellosis]. AB - Because the symptoms and signs of Brucellosis are nonspecific the diagnosis is made with certainty by isolation of brucella or of its nucleic acids, or by evidence of a specific immunologic response. The rate of isolation from blood varies from 50 to 80% during the acute phase; bone marrow cultures may have a higher rate than blood in chronic disease. Recently, the isolation time for brucellae may be reduced by automated culture systems (e.g. the lysis concentration technique or BACTEC) from weeks to days. Although a number of techniques have been developed to measure brucella antibodies (immunoenzymatic, immunofluorescent assays), the serum agglutination test is the simplest and most widely used. Recently, the Polymerase Chain Reaction has been used to evidence nucleic acids of brucella; although it shows high specificity and sensitivity, in the acute and chronic disease, and allows rapid diagnosis, standardization and further evaluation are needed. PMID- 12084985 TI - Glypican-2 binds to midkine: the role of glypican-2 in neuronal cell adhesion and neurite outgrowth. AB - Cell-surface heparan sulfate proteoglycans participate in molecular events that regulate cell adhesion, migration, and proliferation. The present study was performed to elucidate whether glypican-2 plays a role in interactions of neurons with midkine (MK), a heparin-binding neuroregulatory factor. MK bound to heparan sulfate chains of glypican-2 in a manner similar to syndecan-3. Microbeads coated with MK or poly-L-lysine induced clustering of glypican-2 as well as syndecan-3. Substratum-bound MK or poly-L-lysine induced cell adhesion of N2a neuroblastoma cells, while only MK promoted neurite outgrowth of these cells. Ligation of cell surface glypican-2 with MK or an antibody against epitope-tagged glypican-2 induced cell adhesion and promoted neurite outgrowth. These results verified that cell-surface glypican-2 bound to MK and suggested that MK-glypican-2 interactions participate in neuronal cell migration and neurite outgrowth. In addition, we observed different localization of epitope-tagged glypican-2 and syndecan-3 on the surface of N2a cells; the result suggested that they may play different roles in MK-mediated neural function. PMID- 12084987 TI - [Potential antibiotic pharmaceutical cost reduction in an operative unit of infectious diseases. Projects for making savings and improvements]. AB - The authors intend to evaluate the results obtained from a pharmaco-economic project included in the 1999 and 2000 budgets. The objectives of the study are: 1) to establish whether there has been a saving in expenses for antibiotic therapy (T.A.) during the first half-year of 1999 and during the first half-year of 2000 and to ascertain the reasons for any positive result; 2) to evaluate the average hospital stay in the first half-year of 2000 and to observe how many times patients were been discharged early with shift therapy. Of the 286 patients (1999) and 309 (2000) considered, we focused attention on 187 (1999) and 190 patients (2000) who were treated with antibiotic therapy. A substantial cost saving was found in antibiotic use (-31%) in the first half-year of 2000 due to the early discharge and the continuation of home therapy for a fair number of patients and due to increased attention in the choice of medicine for less serious diseases. The second objective was not achieved with the reduction of average hospital stay even if the average length of antibiotic therapies (D.M.T.) fell from 13.1 days at 1st September to 9.8 days in the first half-year of 2000 (P:NS). These results suggest that projects designed to achieve financial savings and improvements require more cooperation between clinical U.O. and services that enable hospital stay to be shortened. PMID- 12084988 TI - Apoptosis and inflammation in renal reperfusion injury. AB - Ischemia followed by reperfusion (I/R) has cardinal implications in the pathogenesis of organ transplantation and rejection. Apoptosis and inflammation are central mechanisms leading to organ damage in the course of renal I/R. General aspects of apoptosis, morphology, induction, and biochemistry are discussed. Activated caspases, the classical effector enzymes of apoptosis, are able to induce not only apoptosis but also inflammation after I/R in experimental models. This redefines the involvement of apoptosis in I/R injury toward a central and functional role in the development of organ damage. Our purpose is to assess aspects of apoptosis and inflammation in terms of involvement in the pathogenesis of I/R-induced organ damage. Moreover, the implications of recent experimental advances for diagnosis and treatment of renal I/R injury in clinical practice will be discussed. PMID- 12084989 TI - Living-related liver transplantation from the view of the donor: a 1-year follow up survey. Transplantation 2002; 73: 1799. PMID- 12084990 TI - VEGF-A and -C but not -B mediate increased vascular permeability in preserved lung grafts. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is a potent endothelial cell growth and permeability factor, expressed in the lung. Overexpression of VEGF is associated with increased vascular permeability in the early stage of acute lung injury in mice. The role of various forms of VEGF in transplantation induced lung injury is not well understood. METHODS: VEGF mRNA and protein expression was measured in biopsies of preserved donor lung grafts as well as in control lung biopsies, using real-time reverse transcriptase-polymerase chain reaction and Western blot analysis. VEGF tissue expression was also evaluated by immunocytochemistry. Serum VEGF was measured in recipients after transplantation and in controls using ELISA. RESULTS: Although VEGF-A and VEGF-C protein expression was up-regulated, their mRNA levels were decreased in donor versus control lung biopsies (P<0.05). VEGF-B mRNA was decreased, but its protein level was unchanged in donors. Flt-1 was unchanged, KDR gene expression was down regulated in donors (P<0.05), and both receptors' protein expression was under the detection level in donor and control lungs. VEGF-A was detected in pulmonary vessels and bronchi, whereas VEGF-C was only detectable in vessels of both donor and control lungs. After transplantation, serum VEGF increased (P<0.05) and returned to control baseline levels 12 weeks after surgery. Wet-to-dry lung weight was increased in donor versus control lungs. CONCLUSIONS: These results indicate that unventilated hypoxia increases vascular permeability in lung grafts and that this process is mainly regulated at VEGF-A and VEGF-C translational but not transcriptional level. Selective VEGF antagonism during graft preservation might be of benefit to counteract edema formation. PMID- 12084991 TI - Mast cells in ongoing acute rejection: increase in number and expression of a different phenotype in rat heart transplants. AB - BACKGROUND: Mast cells (MC) are resident in healthy hearts and play important physiological and pathophysiological roles. In the transplanted heart, correlations have been found between MC number and the severity of rejection episodes, the intensity of chronic inflammation, and allograft arteriosclerotic changes. However, not much emphasis has been placed on the fact that resident donor MC, and infiltrating recipient MC do not forcedly need to share the same properties and function. To gain insight in the role of cardiac MC during acute, and ongoing acute rejection of heart transplants, we investigated MC kinetics and MC phenotype in a rat heart transplantation model. METHODS: Donor hearts from female Brown-Norway rats were transplanted to male Lewis rats. Immunosuppression was started at day 5 using ciclosporin and prednisolone. Connective tissue type MC (CTMC) were distinguished from mucosa type MC (MMC) by immunohistochemistry for rat MC protease (RMCP) -1 and -2. Expression of RMCP-1 and -2 mRNA was quantified by real time reverse transcription-polymerase chain reaction. Infiltrating Y chromosome positive MC were detected by fluorescence in situ hybridization. mRNA expression of interleukin-3 (IL-3) and of the two differentially spliced isoforms of kit ligand (KL, stem cell factor) was quantified using reverse transcription-polymerase chain reaction. RESULTS: Resident cardiac donor MC are almost exclusively CTMC and decrease in number during acute rejection. MC increase in number, and recipient MC invade the cardiac allograft during ongoing acute rejection. The phenotype of the invading MC is characterized by the expression of RMCP-2, or both RMCP-1 and RMCP-2, and thus resemble a MMC type. IL-3 mRNA is highly expressed, and the ratio of the differentially spliced mRNAs for KL-1 and KL-2 rises up to 2-fold during ongoing acute rejection. CONCLUSIONS: Our data show that MC in posttransplant hearts during ongoing acute rejection differ from MC in healthy hearts and isografts by expressing a different phenotype. Changes in IL-3 and KL expression might be responsible for the predominance of MMC over CTMC. The notion is of importance that MC in cardiac allografts may have properties and functions that differ from those in nontransplanted healthy hearts. PMID- 12084992 TI - Disseminated intravascular coagulation in association with pig-to-primate pulmonary xenotransplantation. AB - BACKGROUND: Profound coagulopathy has been proposed as a barrier to xenotransplantation. Disseminated intravascular coagulation (DIC) has been observed with the rejection of renal and bone marrow xenografts but has not yet been described in pulmonary xenografts. METHODS: This study examined the coagulation parameters in five baboons that received pulmonary xenografts and one baboon that was exposed to porcine lung during an extracorporeal perfusion. Platelet counts, prothrombin times (PT), and levels of fibrinogen, D-dimers, and thrombin-antithrombin III complex (TAT) were analyzed. In addition, serum levels of plasminogen activator inhibitor-1 (PAI-1), thrombomodulin (TM), tissue plasminogen activator (tPA), and tissue factor (TF) were determined. RESULTS: Hyperacute pulmonary xenograft dysfunction, which occurred within 0-9 hr of graft reperfusion, was associated with clinically evident DIC. This coagulopathy was characterized by thrombocytopenia, decreased fibrinogen levels, elevations in PT, and increases in D-dimers and TAT. Furthermore, transient increases in PAI-1, increases in TM, and increases in tPA were observed in the serum of some but not all recipients. None of the baboons demonstrated measurable increases in soluble TF. CONCLUSIONS: Although DIC in renal or bone marrow xenotransplantation develops over a period of days, DIC associated with hyperacute pulmonary xenograft dysfunction develops within hours of graft reperfusion. Thus, the DIC in pulmonary xenotransplantation may represent a unique and/or accelerated version of the coagulopathy observed with renal and bone marrow xenotransplantation. PMID- 12084993 TI - Nitric oxide in the development of obliterative bronchiolitis in a heterotopic pig model. AB - BACKGROUND: Inflammation, epithelial cell injury, and development of fibrosis and airway obliteration are the major histological features of posttransplant obliterative bronchiolitis (OB). The expression of inducible nitric oxide synthase (iNOS) in the damaged epithelium, accompanied by peroxynitrite, suggests that endogenous nitric oxide (NO) mediates the epithelial destruction preceding obliteration. To elucidate the role of NO in this cascade, heterotopic bronchial allografts were studied in pigs. METHODS: Allografts or autografts were harvested serially 3-90 days after transplantation and processed for histology and immunocytochemistry for iNOS, nitrotyrosine, a marker of peroxynitrite formation, and superoxide dismutase (SOD). RESULTS: During initial ischemic damage to the epithelium, iNOS, nitrotyrosine, and SOD were found to be strongly expressed in the epithelium of all implants as well as later, after partial recovery, parallel to onset of epithelial destruction and subsequent airway obliteration in allografts. The levels of expression of iNOS in fibroblasts during the early phase of obliteration paralleled the onset of fibrosis. Constant expression of iNOS and SOD, but not nitrotyrosine, occurred in autografts and allografts with blocked alloimmune response. CONCLUSIONS: These findings suggest that an excessive amount of NO promotes posttransplant obliterative bronchiolitis by destroying airway epithelium and stimulating fibroblast activity. SOD may provide protection by binding reactive molecules and preventing peroxynitrite formation. PMID- 12084994 TI - Effect of cold-ischemia time on C-X-C chemokine expression and neutrophil accumulation in the graft liver after orthotopic liver transplantation in rats. AB - BACKGROUND: The precise mechanisms leading to polymorphonuclear neutrophil (PMN) recruitment and activation in the extended cold-preserved liver after transplantation are not yet fully understood. METHODS: We histologically evaluated the number of accumulated PMNs in graft livers, with varying time periods of cold ischemia (1, 6, and 24 hr in University of Wisconsin solution at 4 degrees C), after liver transplantation in rats. Intragraft expression of macrophage inflammatory protein-2 (MIP-2) and cytokine-induced neutrophil chemoattractant (CINC) mRNA, as well as immunohistochemical expression of MIP-2 and CINC in the graft liver, were investigated after reperfusion. The levels of MIP-2 and CINC in the hepatic vein, and tumor necrosis factor (TNF)-alpha, which stimulates these chemokine production, were also monitored. RESULTS: The number of accumulated PMNs in sinusoids significantly increased in the 24-hr cold ischemia group within 3 hr after reperfusion, compared with the 1-hr and 6-hr groups. Serum MIP-2 levels in the 24-hr group significantly increased at 3, 6, and 12 hr after reperfusion, compared with the other groups. Intragraft MIP-2 mRNA was also up-regulated to a greater extent in the 24-hr group. Similarly, serum CINC levels in the 24-hr group significantly increased at 3 hr, compared with the 1-hr group. CINC mRNA also increased as cold-ischemia time was prolonged. Immunohistochemical staining revealed that hepatocytes were the main source of both MIP-2 and CINC protein. In addition, TNF-alpha in the hepatic vein was detected only in the 24-hr group after reperfusion. CONCLUSION: Extended cold preservation of the graft liver might up-regulate MIP-2 and CINC expression of hepatocytes, most probably through elevated TNF-alpha, and might contribute to PMN recruitment and activation after reperfusion. PMID- 12084996 TI - Enhanced development of caspase-independent cortical cell death during cold storage in kidneys of non-heart-beating donors. AB - BACKGROUND: Understanding the mechanisms of injury associated with cardiac arrest is essential for defining strategies aimed at improving preservation and function of kidneys harvested in non-heart-beating (NHB) donors. METHODS: We standardized a model of NHB donors in rats and studied the kinetics and types (apoptosis vs. necrosis) of renal cell death developing during cold storage. Using quantitative polymerase chain reaction, immunoblotting, and caspase inhibition, we also studied the molecular pathways regulating renal cell death in this model. RESULTS: The kinetics and extent of cell death developing in cortical tubules during cold storage were found to be increased in non-heart-beating (NHB) kidneys. Apoptosis of cortical tubules predominated in NHB kidneys exposed to 10 hr of cold storage, whereas necrosis increased after longer periods of cold ischemia. Shortly after cardiac arrest, a rapid up-regulation of Bax and Hsp 70 was found at the protein level in NHB kidneys. After 24 hr of cold storage, induction of Bax was maintained, whereas protein levels of Hsp70 returned to levels comparable to heart-beating (HB) controls. Also, mRNA levels of Bax were found to increase during cold storage in NHB kidneys. Cortical cell death was found to be largely caspase-independent but responsive to hydroxyl-radical scavenging with dimethyl sulfoxide (DMSO). CONCLUSIONS: Cardiac arrest promotes activation of death-inducing molecules such as Bax and is associated with increased development of caspase-independent renal cell death during cold storage. Developing strategies, such as free radical scavenging, aimed at inhibiting cell death during cold storage, could prove useful for improving preservation of NHB kidneys. PMID- 12084995 TI - A novel CD154 monoclonal antibody in acute and chronic rat vascularized cardiac allograft rejection. AB - BACKGROUND: The CD40-CD154 interaction is critically important in the cell mediated immune responses. Blockade of this costimulatory pathway has been shown to prevent acute allograft rejection in murine, as well as nonhuman primate models. However, the role of the CD40-CD154 pathway in the development of chronic rejection and the effects of CD154 targeting on progression of chronic rejection have not been evaluated. METHODS: We examined the effect of AH.F5, a new hamster anti-rat CD154 monoclonal antibody, in a fully allogeneic acute(u) into Lewis [LEW] (RT11) and chronic [WF.1L (RT1l) into LEW (RT1l)] vascularized cardiac allograft rejection model. In the chronic model, the antibody was evaluated for prevention (starting day of transplant) and interruption of progression (starting day 30 or 60 after transplant) of chronic vasculopathy. Graft survival, morphology, and immunohistology were evaluated. RESULTS: In the acute rejection model, anti-CD154 therapy alone prevented acute allograft rejection and resulted in 50% long-term allograft survival (>200 days) and donor-specific tolerance. In recipients treated with anti-CD154 monoclonal antibody in combination with a short course of cyclosporine, 100% of allografts survived long-term and all recipients achieved donor-specific tolerance. In the chronic rejection model, allografts from animals treated with the anti-CD154 antibody had a statistically significant lower score of graft arteriosclerosis and fibrosis in both the prevention and 30-day interruption groups when compared with control allografts. In addition, immunohistochemistry showed a decrease in intragraft mononuclear cell infiltration and activation. CONCLUSION: A new anti-CD154 antibody not only prevents acute allograft rejection, but also inhibits and interrupts the development of chronic rejection. In the acute rejection model cyclosporine acts synergistically with anti-CD154 therapy to prolong allograft survival and induce tolerance. In the chronic rejection model relatively early initiation of therapy is essential to prevent progression of chronic allograft vasculopathy and fibrosis. PMID- 12084997 TI - Reconstructed human skin produced in vitro and grafted on athymic mice. AB - BACKGROUND: The best alternative to a split-thickness graft for the wound coverage of patients with extensive burns should be in vitro reconstructed autologous skin made of both dermis and epidermis and devoid of exogenous extracellular matrix proteins and synthetic material. We have designed such a reconstructed human skin (rHS) and present here its first in vivo grafting on athymic mice. METHODS: The rHS was made by culturing newborn or adult keratinocytes on superimposed fibrous sheets obtained after culturing human fibroblasts with ascorbic acid. Ten days after keratinocyte seeding, reconstructed skins were either cultured at the air-liquid interface or grafted on athymic mice. We present the macroscopic, histologic, and phenotypic properties of such tissues in vitro and in vivo after grafting on nude mice. RESULTS: After maturation in vitro, the reconstructed skin exhibited a well developed human epidermis that expressed differentiated markers and basement membrane proteins. Four days after grafting, a complete take of all grafts was obtained. Histological analysis revealed that the newly generated epidermis of newborn rHS was thicker than that of adult rHS after 4 days but similar 21 days after grafting. The basement membrane components (bullous pemphigoid antigens, laminin, and type IV and VII collagens) were detected at the dermo-epidermal junction, showing a continuous line 4 days after grafting. Ultrastructural studies revealed that the basement membrane was continuous and well organized 21 days after transplantation. The macroscopic aspect of the reconstructed skin revealed a resistant, supple, and elastic tissue. Elastin staining and elastic fibers were detected as a complex network in the rHS that contributes to the good elasticity of this new reconstructed tissue. CONCLUSIONS: This new rHS model gives supple and easy to handle skins while demonstrating an adequate wound healing on mice. These results are promising for the development of this skin substitute for permanent coverage of burn wounds. PMID- 12084999 TI - Platelet adherence to isolated rat hepatic sinusoidal endothelial cells after cold preservation. AB - BACKGROUND: Platelet adhesion to hepatic sinusoidal endothelial cells (SEC) is a major mechanism of cold preservation injury. This study was performed to determine whether cold preservation leads to adhesion between SEC and platelets in the absence of other cell types, whether adherent platelets become activated upon adhesion, and whether there is increased expression of the platelet receptor von Willebrand factor (vWF) on cold preserved SEC. Because we previously showed that cold causes actin disassembly and matrix metalloprotease (MMP) secretion by SEC, we also questioned whether these events are related to increased adhesion of platelets to SEC after preservation. METHODS: Isolated SEC were cold preserved for 8 hr and rewarmed briefly. Biotinylated platelets were added to the plates and adhesion was assessed. Activation of platelets was determined by staining with antibody to P-selectin. Expression of vWF was assessed with a specific antibody. RESULTS: Cold preservation induced increased adhesion of platelets to SEC in the absence of other cell types. Adherent platelets were activated. Preservation increased the expression of vWF on SEC. Pretreatment with phalloidin or treatment with MMP inhibitors partly prevented platelet adhesion and activation, as well as vWF expression. Treatment of SEC at 37 degrees C with recombinant human MMPs for 24 hr also rendered the SEC more adherent for unactivated platelets. CONCLUSION: Cold preservation of SEC results in increased expression of the platelet receptor vWF, increased platelet adhesion, and platelet activation. This is at least partly mediated by actin disassembly and MMP secretion. PMID- 12084998 TI - Effect of mixed hematopoietic chimerism on cardiac allograft survival in cynomolgus monkeys. AB - BACKGROUND: We have previously reported the successful induction of mixed chimerism and long-term acceptance of renal allografts in MHC-mismatched nonhuman primates after nonmyeloablative conditioning and donor bone marrow transplantation. In this study, we extended our regimen to cardiac allotransplantation and compared the immunological responses of heart and kidney allograft recipients. METHODS: Five cynomolgus monkeys were conditioned with low dose total body irradiation (1.5 Gy on days -6 and -5), supplemental thymic irradiation (7 Gy on day -1), antithymocyte globulin (50 mg/kg on days -2, -1, and 0), splenectomy (day 0), donor bone marrow transplantation (day 0), and a 4 week posttransplant course of cyclosporine. Heart allografts from MHC-mismatched donors were transplanted heterotopically on day 0. RESULTS: Two monkeys failed to develop multilineage chimerism and rejected their allografts soon after cyclosporine was stopped (postoperative days [PODs] 43 and 56). Three monkeys developed multilineage chimerism, which persisted 20 to 43 days posttransplant by flow cytometric analysis and to POD 124 by polymerase chain reaction analysis. Allograft survival in these recipients was prolonged to 138, 428, and 509 days, and in vitro mixed leukocyte reaction and cell-mediated lympholysis (CML) assays demonstrated donor-specific hyporesponsiveness. However, in contrast to kidney allograft recipients, long-term heart allograft recipients eventually developed humoral and cellular immunity against the donor and rejected the grafts. At the time of rejection, 1.3% to 9.5% of donor coronary arteries exhibited intimal proliferation. CONCLUSIONS: The induction of transient mixed hematopoietic chimerism leads to long-term heart allograft survival in MHC disparate monkeys without chronic immunosuppression. However, unlike kidney allografts, full tolerance to cardiac allografts was not achieved. Organ-specific modifications of the preparative regimen may be necessary to prevent the chronic cellular and humoral immune responses elicited by cardiac allografts. PMID- 12085000 TI - Antisense deoxyoligonucleotides or antibodies to murine MD-1 inhibit rejection of allogeneic and xenogeneic skin grafts in C3H mice. AB - BACKGROUND: Altered expression of murine MD-1, a molecule controlling expression of members of the interleukin (IL)-1 receptor family of signaling proteins, regulates antigen-presenting cell-induced alloreactions. We investigated the effect of treatment with antisense deoxyoligonucleotides or antibodies to MD-1 in vivo on allogeneic and xenogeneic skin graft survival and the immune responses in transplanted mice. METHODS: C3H mice received C57BL/6 or Lewis rat skin grafts, followed by i.v. injections of anti-MD-1 antibody or antisense oligonucleotides or control reagents at 48-hr intervals. Survival was monitored. In separate studies, mice were sacrificed at 5-day intervals. Serum was analyzed for circulating MD-1 antigen, and peritoneal cells for surface expression of MD-1. The proliferative and cytolytic response of lymphocytes harvested from treated animals and restimulated in vitro with allo- or xenogeneic cells, and the cytokines produced, was measured. Graft histology was assessed at 11 days after transplantation. RESULTS: Treatment with anti-MD-1 oligonucleotides or antibodies suppressed rejection of both xeno- and allogeneic grafts, decreased induction of graft-specific cytotoxic T cells, increased production of type-2 cytokines (IL-4 and IL-10), and decreased production of type-1 cytokines (IL-2 and interferon gamma). Serum levels of MD-1 were suppressed, as was expression of MD-1 on the surface of antigen-presenting cells. Grafts from MD-1-treated mice showed little lymphocyte infiltration, and no signs of graft necrosis. CONCLUSION: Our data suggest a critical in vivo role for MD-1 expression in regulating graft rejection, as well as in the concomitant sensitization of T cells and their cytokine production profile, which parallels the rejection response. PMID- 12085001 TI - Analysis of the control of the anti-gal immune response in a non-human primate by galactose alpha1-3 galactose trisaccharide-polyethylene glycol conjugate. AB - BACKGROUND: The current limitation to the clinical application of xenotransplantation using pig organs is a rejection process that has been termed delayed xenograft rejection or acute vascular rejection. It is thought that acute vascular rejection may be mediated at least in part by both the continued synthesis, of preexisting, and the induction, posttransplantation, of antibodies against the carbohydrate moiety galalpha1-3gal that is present on glycoproteins and glycolipids of the pig endothelium. The synthesis of these antibodies has proven difficult to control with currently available immunosuppressive agents. METHODS: We have synthesized galalpha1-3gal conjugated polyethylene glycol polymers that can bind to anti-galalpha1-3gal antibodies and tested their activity in non-human primates. RESULTS: These conjugates when administered to non-human primates can substantially reduce the levels of preexisting and control the induction of anti-galalpha1-3gal antibodies. The level of circulating antibody-secreting cells that make anti-galalpha1-3gal antibodies is also reduced. CONCLUSION: These alpha-gal polyethylene glycol conjugates may have the potential to control the anti-gal antibody response in a pig to primate organ transplant setting and may be a useful therapeutic agent in prolonging graft survival. PMID- 12085002 TI - Bone mineral density and height gain in children with chronic cholestatic liver disease undergoing transplantation. AB - BACKGROUND: Osteodystrophy is a well-described complication of chronic liver disease. Previous reports in adults and children undergoing liver transplantation (LT) were discordant, with the former showing no improvement of bone disease in the first year after transplantation and the latter demonstrating remarkable benefit from it. Our aim was to perform a pilot study on osteodystrophy in children undergoing LT and evaluate the contribution of growth on bone mineral density (BMD) changes. METHODS: We studied six patients (two male), with a median age at transplantation of 8.8 (range 3.8-16.6) years. Indications for transplantation were biliary atresia and progressive familial intrahepatic cholestasis (three patients each). BMD was studied with dual-energy x-ray absorptiometry and biochemical markers of liver and bone function in patients before and at 3, 6, and 12 months after LT. RESULTS: Median L2-L4 spinal BMD was 0.54 g/cm2 (range 0.29-0.87) before LT, and 0.58 g/cm2 (0.27-0.86) at 3 months, 0.66 g/cm2 (0.36-1.00) at 6 months, and 0.76 g/cm2 (0.44-1.02) at 12 months after LT (P=0.005). Median height was 133 (range 93-167) cm before LT, and 134 (93-167) at 3 months, 136 (97-167) at 6 months, and 139 (102-167) at 12 months after LT. There was direct correlation between height gain and total body BMD improvement (r=0.929, P=0.007). CONCLUSION: BMD in children with chronic cholestatic liver disease improves remarkably by 12 months after LT. Catch-up growth in children can account for the different effect of LT on bone density between adult and pediatric populations in the first year after surgery. PMID- 12085003 TI - Addition of inhaled corticosteroids to systemic immunosuppression after lung transplantation: a double-blind, placebo-controlled trial. AB - BACKGROUND: It is postulated that bronchiolitis obliterans syndrome (BOS) is preceded by airway inflammation that has been described even in stable lung transplant recipients. Airway inflammation is known to be suppressed by inhaled steroids in other chronic inflammatory lung diseases, e.g., asthma and chronic obstructive pulmonary disease. BOS is the major cause of morbidity and mortality after lung transplantation. OBJECTIVE: To examine the effect of inhaled corticosteroids on the development of BOS in lung transplant recipients. METHODS: Thirty patients were recruited and randomized in a double-blind fashion to receive either 750 microg of fluticasone propionate (FP) or an identical appearing placebo twice daily for 3 months; 20 of this group continued until 2 years posttransplantation. Detailed spirometry was performed regularly throughout the study. RESULTS: In the short-term study no differences were found in any examined parameters. In the long-term component of the study no differences were found in the development of neither BOS nor survival. There were minor differences in bronchoalveolar lavage (BAL) lymphocyte percentages. CONCLUSIONS: FP is ineffective for the prevention of BOS after lung transplantation despite the airway inflammation that characterizes this condition. Inadequate local delivery, timing of the therapy relative to transplantation and inherent steroid resistance of this condition may explain the negative finding of this study. PMID- 12085004 TI - Living-related liver transplantation from the view of the donor: a 1-year follow up survey. AB - BACKGROUND: In the past, follow-up surveys for living-related liver transplantation (LRLT) mainly focused on the medical outcome of recipients and donors. In this survey the prevalence of personal, familial, or economic problems of the donors and changes of quality of life after donation were studied. METHODS: Questionnaires were sent to 24 donors after right hepatectomy for LRLT (response 92%). The modified EUROTOLD (European Multicenter Study of Transplantation Using Living Donors) questionnaire was used to inquire about the decision-making process, family problems, and economic problems related to the donation. Global quality of life was measured with the SF-36 Health Survey. RESULTS: For most donors the decision to donate was easy or not very difficult (21/22) and was made spontaneously (17/22). The amount of information about the risks of LRLT was limited at the time of decision but increased remarkably immediately before the operation. In 28%, family conflicts occurred (5/22). Retrospectively, all but two donors (91%) would donate again. On average, donors started working after 9 (+/-3.7) weeks and felt fully recovered after 13 (+/-7.3) weeks. Adverse financial affects were experienced by 41% of the donors (9/22) because of the donation, and four of those received a compensation. Importantly, quality of life did not differ between donors and nondonors. CONCLUSION: Donors viewed LRLT positively. Quality of life after donation did not change. However, donors had a prolonged period of physical rehabilitation, and 41% experienced financial disadvantages. PMID- 12085005 TI - Indirect alloreactivity and chronic rejection. AB - BACKGROUND: The relative contribution of the direct versus indirect pathway of T lymphocyte alloreactivity to the development of chronic rejection is incompletely understood. Utilizing a murine model of cardiac allograft vasculopathy (CAV) and a recipient strain with markedly reduced capacity for indirect alloreactivity, we sought to define the importance of indirect allorecognition in CAV. METHODS: The cells from H2-M mutant mice are unable to present intact protein antigens via class II molecules and have a markedly reduced capacity to present exogenous peptides. B6C.H-2(bm12) strain donor hearts were transplanted into either C57Bl/6 wild-type (WT) or H2-M mutant mice (on C57Bl/6 background). Recipients were killed on day 24. T lymphocyte and macrophage infiltration were graded immunohistochemically. Intimal lesions were measured morphometrically. RESULTS: Donor hearts in WT recipients developed significant intimal lesions, as expected (50+/-7%). Moreover, the donor hearts in H2-M mutant mice also developed comparable intimal lesions (52+/-9%, P=NS vs. WT). Furthermore, the extent of T lymphocyte and macrophage infiltration was similar in both groups. CONCLUSIONS: This study demonstrates that a markedly reduced capacity for indirect alloreactivity does not affect the severity of intimal lesions in this model of CAV. The findings of this study question the role of indirect alloreactivity as the sole pathway of allorecognition leading to chronic rejection. PMID- 12085006 TI - Tacrolimus and cyclosporine differ in their capacity to overcome ongoing allograft rejection as a result of their differential abilities to inhibit interleukin-10 production. AB - BACKGROUND: Accumulated evidence from clinical transplantation has suggested that tacrolimus-based treatment can reverse ongoing allograft rejection in patients treated with cyclosporine (CsA)-based immunosuppression, even when a high dose of antirejection rescue therapy has failed. This evidence prompted us to investigate whether these two compounds, which share an in vitro mechanism, would differ in their abilities to regulate in situ cellular and molecular events during ongoing allograft rejection. METHODS: The equivalent effective doses of tacrolimus (3.2 mg/kg/day) and CsA (10 mg/kg/day), when administered orally to Lewis rats for 10 days (day 0-9), were predetermined and defined as the ability of the drug to induce a similar survival of Brown Norway rat heart allografts with an equal suppression of intragraft interleukin (IL)-2 mRNA expression. To investigate the ability of each drug to rescue ongoing allograft rejection, Lewis recipients of Brown Norway rat heart grafts were left untreated for the first 5 days after transplantation. Tacrolimus or CsA was then administered at the equivalent effective dose for 10 days (days 5-14). Heart grafts and blood samples, harvested on days 3, 5, 7, and 10, were analyzed by reverse transcriptase-polymerase chain reaction, real-time quantitative polymerase chain reaction, ELISA, and immunohistology. RESULTS: Ongoing allograft rejection was found to be rescued by tacrolimus but not by CsA at the equivalent dose (median survival time: untreated, 6 days; tacrolimus, 18 days; and CsA, 7 days). A significant suppression of local intragraft IL-10 mRNA expression and serum protein production along with a dramatic down-regulation of functional CD8+ T and NKR P1a+ natural killer cell local infiltration by means of decreased of cytotoxic factor release, including granzyme B and perforin 1, was found to be associated with tacrolimus but not CsA treatment. However, both drugs inhibited other immune cells (CD4+ T cell, ED2+ macrophage) and cytokines (IL-1beta, IL-2, IL-4, IL-6, IL-12, interferon-gamma, transforming growth factor-beta, and tumor necrosis factor-alpha) at almost the same levels. The inability of CsA to overcome ongoing allograft rejection could be rescued by cotreating recipients with neutralizing anti-IL-10 antibody on day 5 and day 6 after transplantation: anti-IL-10 antibody alone did not show such an effect. CONCLUSIONS: Inhibition of IL-10 production is a critical factor in the ability of tacrolimus to reverse ongoing allograft rejection. PMID- 12085007 TI - Liver repopulation after cell transplantation in mice treated with retrorsine and carbon tetrachloride. AB - BACKGROUND: Efficiency of engraftment after liver cell transplantation is less than 1% under conventional conditions. Our aim was to develop a high-efficiency, nonsurgical, no-genetic-advantage mouse model of liver repopulation with transplanted cells. METHODS: Mice were conditioned with nonlethal doses of a cell cycle inhibitor, retrorsine, 70 mg/kg, to irreversibly block proliferation of native hepatocytes. After the drug was eliminated, 2 million freshly isolated beta-galactosidase-labeled liver cells were transplanted into the spleens of C57BL/6J recipient mice. To stimulate donor cell proliferation, three doses of carbon tetrachloride (CCl4), 0.5 ml/kg, were given. Several control groups were studied to evaluate the contribution of each treatment to liver repopulation. RESULTS: Repopulation, as measured by cell isolation from recipient livers 1-7 months after transplantation, was on average 20%. Repopulation was 10% if CCl4 was given only once, between 0.5% and 1% if only retrorsine or CCl4 were used, and 0.05% if no conditioning was used. Phenotypically, whole livers turned blue on exposure to X-gal staining, whereas negative (control) livers remained pale brown. More than 55% of liver repopulation resulted from clusters containing 21 or more cells, some of which contained more than 200 cells, suggesting seven or more rounds of cell division in a subset of transplanted cells. CONCLUSION: This murine study demonstrates high levels of repopulation after liver cell transplantation into nongenetically modified livers, using a cell cycle inhibitor and chemical liver injury to provide transplanted cells a proliferative advantage. Liver repopulation was effected mostly by a small fraction of transplanted cells. Analogous nonsurgical liver cell transplantation strategies, but with clinically applicable drugs, could be devised for the treatment of liver based metabolic diseases. PMID- 12085008 TI - MDR1 gene expression in peripheral blood mononuclear cells after liver transplantation. AB - BACKGROUND: The multidrug resistance pump, P-glycoprotein 170 (P-gp), the MDR1 gene expression product, is expressed in peripheral blood mononuclear cells (PBMC). Cyclosporine (CsA) and tacrolimus are inhibitors and substrates of the pump, and can induce its expression. The aim of the present study was to assess MDR1 gene expression in PBMC after liver transplantation. METHODS: P-gp expression was examined in PBMC from 23 adult patients by flow cytometry analysis using UIC2, a monoclonal anti-Pgp antibody, on days 7, 14, 21, 28, 90, and 180, and whenever a rejection episode was suspected. MDR1 mRNA was estimated by reverse transcription-polymerase chain reaction. RESULTS: P-gp expression, assessed as the ratio of the mean fluorescence, increased from 1.59 before transplantation to 2.05 after transplantation, and did not vary significantly thereafter. There was no significant difference in P-gp expression between CsA and tacrolimus-based treatments. MDR1 mRNA levels did not vary significantly under CsA or tacrolimus therapy until day 28. P-gp expression was marginally higher in patients before acute rejection (P=0.054), and was lower in patients with severe infection (P=0.003) than in those with uneventful evolution. Alcoholic patients exhibited higher levels of P-gp expression than other patients. CONCLUSION: P-gp expression increases after liver transplantation. Its variations do not allow the ability to predict acute rejection, but could help in making the diagnosis of overimmunosuppression. PMID- 12085009 TI - Lymphocyte apoptosis and cell replacement in human liver allografts. AB - BACKGROUND: Apoptosis of graft-infiltrating T cells has been described after rodent liver transplantation. The aim of this study was to assess lymphocyte apoptosis in human allografts. Additionally, kinetics of leukocyte turnover were studied to determine whether apoptotic cells were likely to be of donor or recipient origin. METHODS: Liver biopsy specimens (n=36) taken between days 3 and 1855 were stained with terminal deoxynucleotidyl transferase dUTP nick end labeling and anti-CD3 to detect apoptotic lymphocytes. Renal allograft and hepatitis C biopsy specimens served as controls. Donor cell turnover was studied in sex-mismatched grafts using Y-chromosome in situ hybridization to detect recipient cells and double immunostaining for leukocyte phenotyping. RESULTS: T cell apoptosis was prominent in hepatic sinusoids (72% of biopsy specimens) as early as day 3. It ranged from 0% to 18.2% of CD3+ cells (mean 5.28+/-0.82%) and persisted for >14 days, including time points >1 year. There was no difference between biopsy specimens with or without rejection (6.34+/-1.14% and 4.61+/ 1.13%, P=NS). Apoptotic cells in portal tracts were less frequent (33% of biopsy specimens) and less abundant (1.13+/-0.36%, P<0.0001). No lymphocyte apoptosis was seen in renal allograft biopsy specimens or hepatitis C biopsy specimens, indicating that it is a distinctive feature of the liver allograft. Persisting lymphocyte apoptosis even after donor lymphocytes had been replaced suggests that recipient lymphocyte deletion must occur. Donor Kupffer cells persisted for many months. CONCLUSIONS: Our results suggest that the sinusoidal microenvironment promotes recipient lymphocyte apoptosis, which may account for the improved outcome of liver grafts compared with other organ allografts. PMID- 12085010 TI - FKBP12 is the only FK506 binding protein mediating T-cell inhibition by the immunosuppressant FK506. AB - BACKGROUND: FK506-binding proteins (FKBP) are immunophilins that interact with the immunosuppressive drugs FK506 and rapamycin. Several FKBP family members such as FKBP12, FKBP12.6, and FKBP51 are expressed in T cells. It has been speculated that these FKBPs are possibly redundant in the immunosuppressant-induced T-cell inactivation. To determine the pharmacological relevance of multiple FKBP members in the immunosuppressant-induced T-cell inactivation, we have investigated the physiological responses of FKBP12-deficient and FKBP12.6-deficient mutant T cells to the immunosuppressive agent FK506. METHODS: FKBP12-deficient and FKBP12.6 deficient T cells were isolated from genetically engineered FKBP12-deficient and FKBP12.6-deficient mice, respectively. T-cell growth inhibitory assay was used to assess their responses to immunosuppressant FK506 treatments. RESULTS: We found that growth inhibition induced by FK506 is abolished in FKBP12-deficient cells but not in FKBP12.6-deficient cells. CONCLUSIONS: FKBP12 is the only FKBP family member that plays a key role in immunosuppressant-mediated immunosuppression. PMID- 12085011 TI - Hand-assisted retroperitoneoscopic living-donor nephrectomy: initial 10 cases. AB - BACKGROUND: Most living-donor nephrectomies are performed either by transperitoneal laparoscopy, with or without hand assistance, or by retroperitoneal open surgery, with or without video assistance. We therefore started to combine the benefits of these techniques: hand assistance to increase safety and control of the laparoscopic technique, and the retroperitoneal approach to minimize the risk of complications associated with the transabdominal approach. Herein, we report on our first 10 donors nephrectomized with hand assisted retroperitoneoscopy. RESULTS: Only left nephrectomies were performed. One donor had two renal arteries and two donors had retroaortic renal veins. There were no intra- or postoperative complications. Mean operating time was 155 min (110-230 min). CONCLUSIONS: Hand-assisted retroperitoneoscopy in living-donor nephrectomy is a promising new method that could reduce the risks of traditional transperitoneal laparoscopy and should be further evaluated. PMID- 12085012 TI - Utility and limitations of serum creatinine as a measure of renal function in experimental renal ischemia-reperfusion injury. AB - BACKGROUND: Ischemia-reperfusion injury (IRI) is the major cause of delayed graft function in renal allografts. The present study was performed to investigate the validity of serum creatinine (SCr) level as an indicator of postischemic renal dysfunction in mice. METHODS: Renal IRI or sham surgery was induced in C57BL/6 mice, and SCr level and inulin clearance (Cin) were measured between 24 hr and 7 days after ischemia. RESULTS: Cin in IRI mice was reduced 75% at 72 hr after ischemia in association with a nearly threefold increase in SCr level. Cin in IRI mice did not recover between 72 hr and 7 days after ischemia, even though SCr level at 7 days was not different between control and IRI mice. In IRI mice, SCr level measured at 24, 48, and 72 hr after ischemia correlated inversely with Cin measured at 72 hr, but not 7 days, after ischemia. CONCLUSIONS: SCr level in the early postischemic period (24-72 hr) seems to be a valid indicator of early postischemic renal dysfunction, and that renal function remains markedly depressed at 7 days despite suggestion from the SCr value that renal function is improving. PMID- 12085013 TI - Improvement in organ shortage through education. AB - Given the persisting shortage of organs for transplantation, it is time to consider whether the message that is sent to society to encourage donation is unsuitable. This message is mainly based on altruism and solidarity, and it has failed. Patients die because of the lack of cadaver organs that society refuses to offer; paradoxically, society is denying itself a chance of life. The reasons for this enigma are (1) lack of awareness that transplantation is a common practice; (2) the persistence of the "cult of dead body integrity;" and (3) the myths surrounding transplantation.A pathway to solving this organ shortage may be education, to make society understand their need of organs for transplantation and its role to resolve this necessity. It should focus on youth but must also reach adults, through their children receiving this education, or from the media or other sources. It should erase the concept of integrity of the cadaver and establish that using cadaver organs means sharing and guaranteeing a source of health for humanity.Similarly, a new message should stress that "using" cadaver organs means "sharing a source of health for humankind." Rather than "a gift of life" we should convey the idea of "sharing a social right and obligation" or that "My decision today assures my and my family health tomorrow." PMID- 12085014 TI - Brain death impairs coronary endothelial function. AB - BACKGROUND: To characterize the impact of brain death (BD) on endothelial dysfunction after cardiac transplantation we investigated coronary circulation and vasomotor function in a canine model. METHODS: Left ventricular pressure volume data (conductance catheter) and coronary blood flow (CBF) were monitored continuously. Endothelium-dependent vasodilatation after acetylcholine and endothelium-independent vasodilation after sodium nitroprusside were assessed before and 3 hr after BD induction (inflation of a subdural balloon). RESULTS: BD led to an initial hyperdynamic reaction with significant (P<0.05) increase of CBF. After 3 hr, CBF decreased significantly (P<0.05). Although before BD, application of acetylcholine led to a monophasic vasodilatative response, after BD a short mild vasodilatation was followed by a longer vasoconstriction. Endothelium-independent vasodilatation remained unchanged. CONCLUSIONS: BD affects coronary circulation by two means: (1) impairment of CBF to decrease in parallel in afterload with consecutive hemodynamic deterioration and (2) severe endothelial dysfunction that may be a contributing factor to posttransplant outcome. PMID- 12085015 TI - Temporary regression of Merkel cell carcinoma metastases after cessation of cyclosporine. PMID- 12085016 TI - Successful treatment of hepatitis C virus after liver transplantation from an identical twin. PMID- 12085017 TI - Pyruvate improvement of cellular energetics, not simply generating ATP during anoxia. PMID- 12085018 TI - Colonic inertia disorders in pediatrics. PMID- 12085019 TI - Picking up the slack--again. PMID- 12085021 TI - Working at the interface. President's message. PMID- 12085022 TI - Moving forward: a new code of ethics. PMID- 12085023 TI - Brandon/Hill selected list of print nursing books and journals. PMID- 12085024 TI - In response to terror: voices of seasoned empathy. PMID- 12085025 TI - Reducing health disparities in the twenty-first century: opportunities for nursing research. AB - This article highlights selected components of the National Institute of Nursing Research's (NINR) Strategic Plan on Reducing Health Disparities. Building on its long history of supporting research devoted to improving minority health, NINR has expanded its opportunities and efforts related to health disparities and enhancing the research career development of minority investigators. Given the increased emphasis on reducing and, ultimately, eliminating health disparities, nurse researchers are encouraged to explore the various opportunities provided by NINR when addressing health inequalities. PMID- 12085026 TI - A vision for tomorrow: transformational nursing leaders. AB - BACKGROUND: Interviews with 8 transformational nursing leaders are presented. PURPOSE: My purpose was to provide a picture of leadership principles and values as practiced by transformational nursing leaders. METHOD: Interviews were conducted as person-to-person, by telephone, or through e-mail. DISCUSSION: The leaders shared their transformational leadership qualities, ideas of a successful environment, effective change strategies, antecedents for success, and visions for the future of nursing practice. CONCLUSIONS: All of the transformational leaders interviewed strive for excellence, value integrity, shape their environment for success, demonstrate perseverance, attempt to improve the lives of others, possess a genuine love for people, motivate others with inspiration, "invent" the future, and share the path toward self-discovery. PMID- 12085027 TI - Establishing a practice-based research network of advanced practice registered nurses in southern New England. AB - BACKGROUND: Biomedical research focuses on highly controlled clinical trials in academic health centers, where the emphasis is on disease treatment and rehabilitation. The results, therefore, are not readily applicable to patient populations that primary care clinicians encounter in their community practices, where the focus is on health promotion and disease prevention. Although the number of practice-based research networks (PBRNs) is growing in primary care, these networks are governed by the physicians who created them and address research questions reflective of physicians' practices and perspectives. Other primary care clinicians, such as advanced practice registered nurses (APRNs), who routinely provide care for underserved and minority populations lack a forum for studying their practice problems and processes. A PBRN of APRNs in primary care provides a means for this research. SUMMARY: The Advanced Practice Registered Nurses' Research Network in southern New England is the first PBRN established for APRNs. This article describes the development of the network. PMID- 12085028 TI - Recurrent bleeding from peptic ulcer associated with adherent clot: a randomized study comparing endoscopic treatment with medical therapy. AB - BACKGROUND: Endoscopic therapy reduces the recurrence of bleeding from actively bleeding peptic ulcers and those with visible vessels. However, the use of endoscopic therapy for ulcers with adherent clots remains controversial. The purpose of this study was to determine whether removal of clot from an ulcer and endoscopic therapy reduces the frequency of recurrent bleeding. METHODS: Patients with acute upper GI bleeding from peptic ulcers with adherent clots and no active bleeding were enrolled in a multicenter study. At each center patients were stratified for age, use of nonsteroidal anti-inflammatory drugs, and ulcer location, and were randomized to endoscopic or medical management. Endoscopic therapy consisted of injection of the base of the adherent clot with a solution of epinephrine and mechanical removal of the clot. The base of the ulcer and any stigmata of bleeding were then coagulated until cavitation and adequate coagulation were obtained. Patients in both groups received standard medical therapy for peptic ulcer. Patients were evaluated for recurrence of bleeding for 1 month. RESULTS: Fifty-six patients were enrolled. Rates of recurrent bleeding were 34.3% (12/35) in the medical treatment arm versus 4.8% (1/21) in the endoscopic treatment arm (p < 0.02). CONCLUSIONS: In patients with GI bleeding caused by gastric or duodenal ulcers with an adherent clot found on endoscopy, endoscopic therapy with injection of the base of the clot, clot removal, and heat probe coagulation significantly reduces the rate of recurrent bleeding compared with medical therapy alone. PMID- 12085029 TI - Outcome differences after endoscopic drainage of pancreatic necrosis, acute pancreatic pseudocysts, and chronic pancreatic pseudocysts. AB - BACKGROUND: Comparative outcomes after endoscopic drainage of specific types of symptomatic pancreatic fluid collections, defined by using standardized nomenclature, have not been described. This study sought to determine outcome differences after attempted endoscopic drainage of pancreatic fluid collections classified as pancreatic necrosis, acute pseudocyst, and chronic pseudocyst. METHODS: Outcomes were retrospectively analyzed for consecutive patients with symptoms caused by pancreatic fluid collections referred for endoscopic transmural and/or transpapillary drainage. RESULTS: Complete endoscopic resolution was achieved in 113 of 138 patients (82%). Resolution was significantly more frequent in patients with chronic pseudocysts (59/64, 92%) than acute pseudocysts (23/31, 74%, p = 0.02) or necrosis (31/43, 72%, p = 0.006). Complications were more common in patients with necrosis (16/43, 37%) than chronic (11/64, 17%, p = 0.02) or acute pseudocysts (6/31, 19%, p = NS). At a median follow-up of 2.1 years after successful endoscopic treatment (resolution), pancreatic fluid collections had recurred in 18 of 113 patients (16%). Recurrences developed more commonly in patients with necrosis (9/31, 29%) than acute pseudocysts (2/23, 9%, p = 0.07) or chronic pseudocysts (7/59, 12%, p = 0.047). CONCLUSIONS: Successful resolution of pancreatic fluid collections may be achieved endoscopically by an experienced therapeutic endoscopist. Outcomes differ depending on the type of pancreatic fluid collection drained. Further studies of endoscopic drainage of pancreatic fluid collections must use defined terminology to allow meaningful comparisons. PMID- 12085030 TI - Pancreatic stent placement for duct disruption. AB - BACKGROUND: The aim of this study was to identify predictors of outcome after pancreatic duct stent placement for duct disruption. METHODS: Patients were identified from endoscopy databases. Disruption was defined by extravasation of contrast from the pancreatic duct during endoscopic retrograde pancreatography. Data collected included demographic information, imaging studies, management before and outcome after stent placement. Success was defined as resolution of the disruption clinically, on radiologic imaging, and/or at endoscopic retrograde pancreatography. RESULTS: Forty-three patients (23 women, 20 men; mean age 57 years, [SD] 15.2 years) were studied. The etiology of pancreatic duct disruption was acute pancreatitis in 24, chronic pancreatitis in 9, operative injury in 7, and trauma in 3 patients. In 25 patients there was resolution of the disruption, whereas stent therapy was unsuccessful in 16 and the outcome was indeterminate in 2 patients. On univariate analysis, stent positioned to bridge the disruption (p = 0.04) and longer duration of stent therapy (p = 0.002) were associated with a successful outcome. Female gender (p = 0.05) and acute pancreatitis (p = 0.05) were associated with a lack of success. On multivariate analysis, only the bridging stent position remained correlated to outcome. Complications occurred in 4 patients. CONCLUSIONS: A bridging stent is associated with a successful outcome after pancreatic duct stent placement for duct disruption. PMID- 12085031 TI - MR cholangiography versus cholangioscopy for evaluation of longitudinal extension of hilar cholangiocarcinoma. AB - BACKGROUND: The utility of magnetic resonance cholangiography for assessment of longitudinal tumor extension of hilar cholangiocarcinoma was investigated with reference to findings by percutaneous transhepatic cholangioscopy. METHODS: Ninety-nine patients with hilar cholangiocarcinoma underwent both magnetic resonance cholangiography and percutaneous transhepatic cholangioscopy. Longitudinal tumor extension was described with the Bismuth-Corlette classification. Hilar cholangiocarcinoma was classified morphologically into stenotic, diffuse sclerosing, and polypoid types based on selective cholangiographic findings obtained during percutaneous transhepatic cholangioscopy. Agreement between percutaneous transhepatic cholangioscopy and magnetic resonance cholangiography according to the Bismuth-Corlette classification was compared. The degree of agreement between magnetic resonance cholangiography and percutaneous transhepatic cholangioscopy according to each morphologic type was also compared in each subgroup without reference to Bismuth Corlette type. RESULTS: The overall agreement between magnetic resonance cholangiography and percutaneous transhepatic cholangioscopy with regard to Bismuth-Corlette types was 87.9% (kappa = 0.832, p < 0.01). The agreement of magnetic resonance cholangiography for each Bismuth-Corlette type with reference to percutaneous transhepatic cholangioscopy was as follows: type I (n = 18), 16/18 (88.9%); type II (n = 16), 14/16 (87.5%); type IIIa (n = 23), 19/23 (82.6%); type IIIb (n = 14), 14/14 (100%); and type IV (n = 28), 24/28 (85.7%). The overall agreement between magnetic resonance cholangiography and percutaneous transhepatic cholangioscopy for Bismuth-Corlette type according to selective cholangiographic findings was as follows: stenotic type, 58/61 (95.1%, kappa = 0.929, p < 0.01); diffuse sclerosing type, 12/16 (75%, kappa = 0.619, p < 0.01); and polypoid type, 17/22 (77.3%, kappa = 0.696, p < 0.01). CONCLUSION: There is good overall agreement between magnetic resonance cholangiography and percutaneous transhepatic cholangioscopy on longitudinal extension of hilar cholangiocarcinoma. Especially for the stenotic type of hilar cholangiocarcinoma (based on selective cholangiographic findings), magnetic resonance cholangiography may replace percutaneous transhepatic cholangioscopy in the determination of longitudinal tumor extension. For polypoid or diffuse sclerosing types, however, percutaneous transhepatic cholangioscopy is required for accurate evaluation of longitudinal tumor extension. PMID- 12085032 TI - Endoscopic palliation of patients with biliary obstruction caused by nonresectable hilar cholangiocarcinoma: efficacy of self-expandable metallic Wallstents. AB - BACKGROUND: The aim of this study was to evaluate the efficacy of an endoscopically inserted self-expandable metal stent for treatment of biliary obstruction caused by nonresectable hilar cholangiocarcinoma. METHODS: Data on all patients with nonresectable hilar cholangiocarcinoma receiving self expandable metal stents between January 1990 and April 2000 were retrospectively reviewed. Patients with pseudo Klatskin tumors (metastases in the liver hilum) and gallbladder carcinoma were excluded. Early complications (within 30 days of stent placement), early and late stent occlusion, stent patency, and biliary reintervention rates were recorded. RESULTS: Fifty-two Wallstents were inserted in 36 patients, with 7, 13, and 16 having, respectively, Bismuth type I/II, III, and IV lesions. Insertion was successful in 35 of 36 (97%) patients. Complications occurred within the first 30 days after insertion in 5 patients (14%) but did not require biliary reintervention. Thirty-day mortality was 6% (2 deaths). There were 3 early (9%) and 8 late (23%) stent occlusions, bringing the total to 11 patients (31%) who required biliary reintervention after Wallstent insertion. Median Wallstent patency and patient survival were, respectively, 169 days (95% CI [154, 184]) and 147 days (95% CI [106, 188]), with no difference between patients with Bismuth I/II, III, IV type tumors. CONCLUSIONS: Wallstent insertion is safe and feasible, and achieves successful palliation without the need for further biliary reintervention in the majority (69%) of patients with nonresectable hilar cholangiocarcinoma. PMID- 12085033 TI - Detecting cholangiocarcinoma in patients with primary sclerosing cholangitis. AB - BACKGROUND: Primary sclerosing cholangitis is a progressive cholestatic liver disease associated with cholangiocarcinoma. Brush cytology and serum tumor markers (carcinoembryonic antigen, carbohydrate antigen 19-9 [CA19-9]) have been used to diagnose cholangiocarcinoma, but there are few data comparing their effectiveness. METHODS: The effectiveness of brush cytology, carcinoembryonic antigen, and CA19-9 for the diagnosis of cholangiocarcinoma was retrospectively studied by review of patients with primary sclerosing cholangitis. Receiver operator curves were used to identify cutoff points for carcinoembryonic antigen and CA19-9. RESULTS: Of 692 patients with primary sclerosing cholangitis screened, adequate follow-up was obtained in 333, 44 (13%) of whom had a diagnosis of cholangiocarcinoma. Three hundred eighteen brush cytology specimens were obtained in 151 patients; serum carcinoembryonic antigen and CA19-9 levels were obtained in 144 and 55 patients, respectively. The overall sensitivity and specificity of brush cytology were, respectively, 46.4% (95% CI [27.5, 64.5]) and 100% (95% CI [97.2, 100]). A carcinoembryonic antigen >5.2 ng/mL had a sensitivity of 68.0% (95% CI [47.5, 83.9]) and specificity of 81.5% (95% CI [73.9, 87.7]). A CA19-9 >180 U/mL had a sensitivity of 66.7% (95% CI [34.9, 87.7]) and specificity of 97.7% (95% CI [88.2, 99.9]). In the subset of patients in which all 3 tests were obtained, (n = 45, cholangiocarcinoma = 8) the combination of an abnormal carcinoembryonic antigen or CA19-9 had the highest sensitivity: 100% (95% CI [65.1, 100.0]) with a specificity of 78.4% (95% CI [63.1, 89.7]). The combination of a positive brush cytology or an abnormal CA19-9 had a sensitivity and specificity of, respectively, 87.5% (95% CI [50.0, 99.4]) and 97.3% (95% CI [86.2, 99.9]). CONCLUSIONS: Screening patients with primary sclerosing cholangitis for cholangiocarcinoma with CA19-9 and carcinoembryonic antigen is reasonable, but the ideal intervals at which to obtain these tests and the cost-effectiveness require further study. PMID- 12085034 TI - Increased risk of colorectal neoplasia in patients with primary sclerosing cholangitis and ulcerative colitis: a meta-analysis. AB - BACKGROUND: Published data on the risk of colorectal neoplasia in patients with ulcerative colitis with and without primary sclerosing cholangitis are conflicting. A meta-analysis was performed to synthesize available publications and to compare the risk of colorectal neoplasia in patients with ulcerative colitis with and without primary sclerosing cholangitis. METHODS: By using MEDLINE and manual search methods, studies were identified that compared the risk of colorectal neoplasia (dysplasia and carcinoma) in patients with ulcerative colitis with and without primary sclerosing cholangitis. In addition, citations were reviewed in relevant articles and proceedings from gastroenterology meetings, and investigators were contacted when data were incomplete. The summary odds ratio (OR) was then calculated for the risk for patients with ulcerative colitis and primary sclerosing cholangitis of having colorectal neoplasia develop compared with that of patients with ulcerative colitis without primary sclerosing cholangitis. RESULTS: Eleven studies met all eligibility criteria for the meta analysis. Patients with ulcerative colitis and primary sclerosing cholangitis are at increased risk of colorectal dysplasia and carcinoma compared with patients with ulcerative colitis alone; OR 4.79: 95% CI [3.58, 6.41] with the Mantel Haenszel method, and OR 5.11: 95% CI [3.15, 8.29] with the Der Simonian and Laird method. This increased risk is present even when the risk of colorectal carcinoma alone is considered; OR 4.09: 95% CI [2.89, 5.76] and OR 4.26: 95% CI [2.80, 6.48] by using, respectively, the Mantel-Haenszel and the Der Simonian and Laird methods. CONCLUSIONS: Patients with ulcerative colitis and primary sclerosing cholangitis have a significantly higher risk for the development of colorectal neoplasia than patients with ulcerative colitis but not primary sclerosing cholangitis. More intensive colonoscopic surveillance should be considered for patients with ulcerative colitis and primary sclerosing cholangitis. PMID- 12085035 TI - Role of cholecystectomy in preventing recurrent cholangitis. AB - BACKGROUND: Whether cholecystectomy should be performed after an episode of acute cholangitis is still unresolved. The purpose of this study was to analyze the role of elective cholecystectomy in preventing recurrent acute cholangitis in Asian patients. METHODS: Two hundred ten consecutive Asian patients with acute cholangitis caused by choledocholithiasis with coexisting cholelithiasis were studied prospectively. RESULTS: Forty-one patients (19.5%, Group 1) agreed to elective cholecystectomy whereas 169 patients (80.5%, Group 2) did not. Mean (+/- SEM) follow-up for Groups 1 and 2 were, respectively, 110.2 +/- 6.6 and 96.8 +/- 2.9 months. Endoscopic papillotomy was performed in 120 patients, 22 (53.7%) in Group 1 and 98 (58%) in Group 2. Recurrent acute cholangitis developed in 31 patients (14.8%), 9 in Group 1 and 22 in Group 2. There was no significant difference in the Kaplan-Meier estimates of the cumulative probability of occurrence of recurrent acute cholangitis between the 2 groups (p = 0.90). Recurrent acute cholangitis developed in 10 patients (8.3%) who underwent endoscopic papillotomy and in 21 (23.3%) patients who did not. There was a significant difference in the Kaplan-Meier estimates of the cumulative probability of occurrence of recurrent acute cholangitis between the patients with endoscopic papillotomy versus those without endoscopic papillotomy (p = 0.001). CONCLUSION: Cholecystectomy did not prevent recurrent acute cholangitis in Asian patients. In these patients, early endoscopic papillotomy lowered the frequency of recurrent acute cholangitis. PMID- 12085036 TI - Management of gallstone pancreatitis: cholecystectomy or ERCP and endoscopic sphincterotomy. AB - BACKGROUND: Currently, cholecystectomy is recommended for patients with gallstone induced pancreatitis. ERCP with endoscopic sphincterotomy (ES) within 24 to 48 hours is also suggested for the treatment of acute gallstone pancreatitis. The aim of this study was to determine outcome after cholecystectomy versus ES alone in patients with gallstone pancreatitis. METHODS: One hundred seventeen patients with gallstone pancreatitis were included in this prospective observational study. Inclusion criteria were typical abdominal pain; serum amylase level 3 times or greater than normal; and gallbladder stones and a dilated bile duct, with or without stones, by US, CT, or ERCP. RESULTS: Eighty-three patients (Group A) underwent cholecystectomy after initial evaluation including ERCP in 43 (53%) and ERCP with ES in 38 (47%). The remaining 34 (Group B) underwent successful ERCP with ES alone. Mean follow-up was 33 months for Group A and 34 months for Group B. Recurrent gallstone pancreatitis was noted in 2 patients (2.4%) in Group A (bile duct stone in 2, sludge and papillary stenosis in 1), and in 1 patient (2.9%) in Group B. Ten patients in Group B had follow-up US of the gallbladder that showed disappearance of stones in 3. During follow-up, there was no significant difference in the rates of biliary complications (Group A, 3.6% vs. Group B, 11.6%; p = 0.19) or serious complications (pancreatitis, cholecystitis, cholangitis) (Group A, 3.6% vs. Group B, 5.8%). Also, there was no significant difference in procedure-related complications. CONCLUSIONS: Recurrence of pancreatitis after ERCP with ES alone for gallstone pancreatitis is rare. In patients who have undergone ES alone, cholecystectomy should be considered only if there are overt manifestations of gallbladder disease (e.g., biliary pain, cholecystitis, cystic duct obstruction) and not for prevention of recurrent gallstone pancreatitis. Because treatment by ES alone may be associated with a higher risk of biliary complications during follow-up compared with cholecystectomy, these patients may require close surveillance. PMID- 12085037 TI - Hereditary pancreatitis: clinical presentation, ERCP findings, and outcome of endoscopic therapy. AB - BACKGROUND: Hereditary pancreatitis (HP) is an autosomal dominant disorder characterized by the onset of recurrent attacks of acute pancreatitis in childhood and frequent progression to chronic pancreatitis. There are limited data on the role of diagnostic and therapeutic ERCP in patients with HP. The aims of this study were to describe the clinical presentation and findings on pancreatography and to evaluate the role of endoscopic and surgical intervention in patients with HP. METHODS: Twenty-seven consecutive patients with HP were referred for ERCP at two centers in the midwest United States. Medical records and pancreatograms were reviewed. RESULTS: Pancreatography was abnormal in all patients. The majority (70%) had severe (Cambridge Grade III) changes of chronic pancreatitis. Nineteen (70%) patients underwent endoscopic therapy and 8 (30%) surgical intervention. After endoscopic therapy (mean follow-up 32 months), 50% were symptom free, 38% had moderate improvement, and 12% had no or transient improvement in symptoms. After surgery (mean follow-up 32 months), 38% were symptom-free, 25% had moderate improvement, and 12% had transient improvement in symptoms. Two patients treated surgically died of pancreatic cancer. CONCLUSIONS: In the short-term, improving drainage of the pancreatic ductal system by endoscopic means effectively relieves pain and reduces the number of attacks of pancreatitis in the majority of patients with HP. PMID- 12085038 TI - Effects of adherence factors and human bile on bacterial attachment and biliary stent blockage: an in vitro study. AB - BACKGROUND: Bacterial attachment plays an important role in the initiation of biliary sludge formation and stent blockage. In vitro studies were conducted to determine the effects of adherence factors, namely pili and glycocalyx production, and culture media, including brain heart infusion broth, modified Vogel and Bonner medium, and human bile, on the adherence of Escherichia coli to plastic stents. METHODS: Clinical isolates of E coli with different adherence mechanisms, that is, piliated (P+) or nonpiliated (P-), glycocalyx producing (G+) and nonglycocalyx producing (G-), were obtained from clogged stents. Adherence studies were conducted by using the modified Robbins device, and stents were removed at regular intervals to determine the number of attached bacteria/cm(2) with the viable plate count method. Polyethylene stents were used to compare the adherence curves of E coli with different adherence factors in brain heart infusion broth. The effects of different culture media on the adherence of P+G+ E coli to polyethylene stents were determined. In addition, the adherence of P+G+ E coli to different plastics in brain heart infusion broth and human bile was compared. RESULTS: P+G+ E coli adhered better than P-G+ and P-G- E coli to polyethylene stents. Modified Vogel and Bonner medium, which stimulates glycocalyx production, enhanced the attachment of P+G+ E coli, whereas human bile decreased E coli attachment to polyethylene stents, despite an increase in glycocalyx production. There was a difference in adherence of P+G+ E coli to polyethylene, polyurethane, and Teflon stents in brain heart infusion broth, but the differences were nullified in the presence of human bile. CONCLUSIONS: P+G+ E coli with both adherence factors adhere best to plastic stents. Media such as modified Vogel and Bonner medium that stimulate glycocalyx production also enhance bacterial attachment. The toxic effects of bile salts in human bile on the bacteria might alter the adherence mechanism and reduce E coli attachment. PMID- 12085039 TI - Prospective evaluation of hemoclip application with injection of epinephrine in hypertonic saline solution for hemostasis in unstable patients with shock caused by upper GI bleeding. AB - BACKGROUND: It is difficult to arrest severe upper GI bleeding with any of the available hemostatic modalities in unstable patients who are in shock, and the rates of persistent bleeding and mortality in this group remain high. This prospective study evaluated hemoclip application alone and in combination with injection of hypertonic saline solution with epinephrine in this subgroup of patients with GI bleeding. METHODS: Twenty-two patients in shock because of upper GI bleeding were enrolled and divided into 2 groups based on the response of systolic blood pressure to rapid infusion of 1000 mL of lactated Ringer's solution: an unstable shock group, in which systolic blood pressure did not stabilize at greater than 90 mm Hg, and a stable shock group, in which systolic blood pressure stabilized at greater than 90 mm Hg. Emergency endoscopy was performed in both groups; those in the stable group were treated by hemoclip application alone and those in the unstable group were treated by hemoclip application combined with injection of hypertonic saline solution with epinephrine. The following parameters were compared: vital signs on admission and after infusion of lactated Ringer's solution, hemoglobin concentration, endoscopic classification of type and site of bleeding, number of hemoclips required to arrest bleeding, volume of hypertonic saline solution with epinephrine injected, initial hemostatic rate, rate of recurrent bleeding, the need for additional preventive therapy (hemoclip application), and mortality. RESULTS: The rate of initial hemostasis was 92% in the stable shock group and 100% in the unstable shock group. Bleeding did not recur in either group. The volume of packed red cells transfused and the endotracheal intubation rate were significantly greater in the unstable shock group. Preventive application of hemoclips was performed at endoscopic follow-up 12 times in 10 patients in the stable shock group and 9 times in 7 patients in the unstable shock group. There were no deaths in either group. CONCLUSION: Endoscopic injection of hypertonic saline solution with epinephrine combined with hemoclip application provides effective hemostasis in unstable patients in shock caused by severe upper GI bleeding. The hemostatic result is comparable with that achieved by hemoclip application alone in patients with bleeding but less severe shock. PMID- 12085040 TI - Use of detachable snares and elastic bands for endoscopic control of bleeding from large gastric varices. AB - BACKGROUND: Bleeding from gastric varices larger than 2 cm in diameter represents a major limitation for endoscopic hemostasis. METHODS: Endoscopic ligation of gastric varices was performed with detachable snares and elastic bands in 41 patients who had recent bleeding from gastric varices larger than 2 cm in diameter. Gastric varices larger than 2 cm were ligated with detachable snares, and then adjacent small gastric varices were ligated with elastic bands. RESULTS: Among the 41 patients, 10 of 12 patients with active bleeding and 28 of 29 patients with red color signs at initial endoscopy were successfully treated by endoscopic ligation by using detachable snares and elastic bands. Bleeding recurred early (before variceal eradication) in 4 of 38 patients (10.5%). The overall hemostatic rate for endoscopic ligation was 82.9% (34/41). In 33 of 36 patients (91.7%) who underwent repeated ligation treatments, variceal eradication was nearly complete. Mean follow-up in 30 of 33 patients after eradication of varices was 16.4 months (3-32 months), and 29 of 30 did not have recurrent bleeding. During and after ligation there were no serious complications. CONCLUSIONS: Endoscopic ligation therapy with large detachable snares and elastic bands is safe and effective for treatment of large bleeding gastric varices. PMID- 12085041 TI - Low-volume oral colonoscopy bowel preparation: sodium phosphate and magnesium citrate. AB - BACKGROUND: Low-volume oral colonoscopy preparations are easier to tolerate and are gaining popularity at the expense of large-volume lavage solutions. The aims of this two-part study were to compare two oral colonoscopy bowel preparations, sodium phosphate (NaP) and magnesium citrate (Mg), and to assess the effects of timing and dilution of NaP. METHODS: Part 1 included 300 ambulatory patients randomized to receive NaP or Mg on the day before colonoscopy. Patients completed a questionnaire before colonoscopy. The endoscopist, blinded to the type of bowel preparation, rated bowel cleansing and recorded the presence of any rectosigmoid aphthous ulcers. In part 2, a total of 297 patients received NaP diluted into 3 doses 10 minutes apart the evening before and the morning of the day of colonoscopy. This group was compared with the group in part 1 that received NaP in two separate single doses entirely the day before colonoscopy. RESULTS: NaP and Mg were well tolerated. When preparations were taken the day before colonoscopy, Mg achieved better bowel cleansing (p < 0.001). Taking NaP entirely the day before colonoscopy resulted in poor right colon cleansing (27%). In contrast, taking NaP the evening before and morning of colonoscopy improved cleansing in the right colon (p < 0.001). Vomiting occurred less frequently with dilution of NaP. Rectosigmoid aphthous ulcers occurred more often with NaP (5.5%) than Mg (1%), p < 0.01. For patients who, according to questionnaire, had received a previous bowel preparation, NaP and Mg were both preferred over large volume lavage solutions, whereas NaP was preferred over Mg. CONCLUSIONS: When taken entirely the day before colonoscopy, Mg is superior to NaP. Taking NaP the evening before and morning of colonoscopy improved bowel cleansing compared with taking it entirely the day before. Dilution of NaP reduces vomiting. NaP induces rectosigmoid aphthous ulcers more often than Mg. PMID- 12085042 TI - Efficacy of colonic submucosal saline solution injection for the reduction of iatrogenic thermal injury. AB - BACKGROUND: Submucosal saline solution injection may limit the depth of thermal injury to the gut wall by acting as a heat-sink and by increasing the distance between burn and serosa. The aim of this study was to determine the effect of submucosal saline solution injection on depth of colonic thermal injury produced by commonly used endoscopic thermal modalities. METHODS: Longitudinal colotomy incisions were made on the antimesenteric colonic border of anesthetized swine. Lesions were made by using a bipolar device (20 W, 2 seconds), heat probe (30 J); monopolar contact with hot biopsy forceps (20 W, 2 seconds), and monopolar noncontact with argon plasma coagulation (45 W, 3 seconds). Ten or more lesions were created with each device. Lesions were made with or without prior submucosal injection of 2 mL of normal saline solution. After 24 hours the lesions were excised for histologic analysis. Injury was assessed in relation to the severity of damage to the deep (longitudinal) muscle layer. RESULTS: The proportions of control lesions (without submucosal saline solution injection) in which deep injury was evident were as follows: argon plasma coagulation, 86%; hot biopsy forceps, 64%; heat probe, 50%; bipolar device, 18%. Submucosal saline solution injection significantly reduced the proportions of lesions with deep injury for argon plasma coagulation (p = 0.009) and heat probe (p = 0.03), but not hot biopsy forceps or bipolar device (argon plasma coagulation, 86% to 21%; heat probe, 50% to 0%; hot biopsy forceps, 64% to 50%; bipolar device, 18% to 9%). CONCLUSIONS: At equivalent energy outputs, the bipolar device results in less deep injury than the monopolar or heat probe. Submucosal saline solution injection reduced injury to the muscularis propria caused by both heat probe and argon plasma coagulation, but not hot biopsy forceps. Despite submucosal saline solution injection, caution should be exercised when using prolonged monopolar cautery. PMID- 12085043 TI - EUS for rectal disease. PMID- 12085044 TI - Infection with a black Pseudoterranova decipiens. PMID- 12085045 TI - Primary ileal mucinous adenocarcinoma. PMID- 12085046 TI - Bouveret's syndrome. PMID- 12085047 TI - Surgical clip choledocholithiasis. PMID- 12085048 TI - Sclerosing cholangitis caused by Clonorchis sinensis. PMID- 12085049 TI - Bleeding Dieulafoy's lesion. PMID- 12085050 TI - A novel endoscopic full-thickness plicator for treatment of GERD: an animal model study. AB - BACKGROUND: This study was designed to demonstrate the efficacy and safety of a novel endoscopic full-thickness plication device for the treatment of GERD. METHODS: Gastric pressures and reflux from the gastroesophageal junction were measured in 20 excised pig stomachs before and after full-thickness plication within 1 cm of the gastroesophageal junction. Endoscopic full-thickness plication was performed on 11 live mini-swine that were subsequently observed for up to 12 weeks. OBSERVATIONS: Full-thickness plication significantly increased the mean intragastric pressure at which gastroesophageal reflux occurred. No complication, death, or sign of infection or tissue damage was noted by endoscopic observation or inspection of excised tissue at up to 12 weeks' follow-up. CONCLUSIONS: These studies in the ex vivo porcine model and live pigs suggest that full-thickness plication is effective and safe. PMID- 12085051 TI - Laryngeal mask airway--a novel method of airway protection during ERCP: comparison with endotracheal intubation. AB - BACKGROUND: ERCP sometimes requires deep sedation and rarely general anesthesia with airway protection. The laryngeal mask airway device is placed perorally to create a seal over the larynx. Unlike endotracheal intubation, no tube traverses the vocal cords, thus reducing airway stimulation and obviating the need to administer muscle relaxants. The feasibility of using the laryngeal mask airway during ERCP was evaluated and recovery times compared for patients undergoing ERCP with the laryngeal mask airway versus endotracheal intubation. METHODS: In this retrospective cohort study, anesthesia records were reviewed for anesthesiologist-assisted ERCP procedures performed during a 30-month period. Demographics, procedure duration, and time from endoscope removal to extubation were abstracted. Either propofol or inhalation agents were used for anesthesia in all patients. OBSERVATIONS: Anesthesiologists administered sedation for 41 ERCPs. The airway was managed in 12 patients with endotracheal intubation and the laryngeal mask airway in 20 patients. Six patients underwent laryngeal mask airway insertion and removal while prone. A therapeutic duodenoscope was passed beyond the laryngeal mask airway with little or no resistance in all cases. Repositioning the laryngeal mask airway during the procedure was required in 1 case. Laryngeal mask airway use was associated with shorter extubation time compared with endotracheal intubation (7.2 vs. 12 min.; p = 0.004). There were no airway complications. CONCLUSION: ERCP can be performed while using the laryngeal mask airway for airway protection. The laryngeal mask airway can be placed with the patient prone, obviating the need to change position. Laryngeal mask airway shortens extubation time compared with endotracheal intubation. PMID- 12085052 TI - EUS-guided pancreaticogastrostomy. AB - BACKGROUND: The pain of chronic pancreatitis can be caused by pancreatic ductal hypertension, and endoscopic drainage of the main pancreatic duct can provide relief. When transpapillary access to a dilated portion of the main duct cannot be obtained, conventional endoscopic drainage is not possible. The use of interventional EUS to perform a pancreaticogastrostomy in such cases is described. METHODS: Four patients presented with pain and a dilated main pancreatic duct proximal to a complete obstruction. EUS was used to access the dilated duct and create a pancreaticogastrostomy. Patency of the latter was maintained by placement of a pancreaticogastric stent. OBSERVATIONS: EUS-guided pancreaticogastrostomy was performed without major complication. Three of 4 patients had satisfactory relief of pain at a median follow-up of 1 year. CONCLUSION: EUS-guided pancreaticogastrostomy may be a promising new technique for pancreatic drainage and pain relief when conventional transpapillary access to the pancreatic duct is not possible. PMID- 12085053 TI - Spontaneous intramural rupture of the esophagus: clinical presentation and endoscopic findings. PMID- 12085054 TI - EUS in combination with fine-needle injection celiac plexus neurolysis from within a Wallstent stent. PMID- 12085056 TI - Esophageal bronchogenic cyst successfully excised by endoscopic mucosal resection. PMID- 12085055 TI - Aortic valve replacement: a last resort for aortic stenosis-associated refractory GI bleeding. PMID- 12085057 TI - Stridor: a complication of PEG placement. PMID- 12085058 TI - Intestinal bleeding due to a stromal tumor in a Meckel's diverticulum. PMID- 12085059 TI - Pancreatic endotherapy for pseudocysts and fluid collections. PMID- 12085060 TI - Hemostasis of upper GI bleeding. PMID- 12085063 TI - Colonoscopically applied clips to treat postoperative fecal fistula. PMID- 12085062 TI - False-negative rapid urease test in acute upper GI bleeding: is human plasma alone responsible? PMID- 12085064 TI - Combined antegrade and retrograde dilation: a new endoscopic technique in the management of complex esophageal obstruction. PMID- 12085066 TI - A study of the workforce in emergency medicine: 1999. AB - STUDY OBJECTIVE: We estimate the total number of physicians practicing clinical emergency medicine during a specified period, describe certain characteristics of those individuals to estimate the total number of full-time equivalents (FTEs) and the total number of individuals needed to staff those FTEs, and compare the data collected with those data collected in 1997. METHODS: Data were gathered from a survey of a random sample of 2,153 hospitals drawn from a population of 5,329 hospitals reported by the American Hospital Association as having, or potentially having, an emergency department. The survey instrument addressed items such as descriptive data on the institution, enumeration of physicians in the ED, and the total number of physicians working during the period from June 6 to June 9, 1999. Demographic data on the individuals were also collected. RESULTS: A total of 940 hospitals responded (a 44% return rate). These hospitals reported that a total of 6,719 physicians were working during the specified period, or an average of 7.85 persons scheduled per institution. The physicians were scheduled for a total of 347,702 hours. The average standard for FTE was 40 clinical hours per week. This equates to 4,346 FTEs or 5.29 FTEs per institution. The ratio of persons to FTEs was 1.48:1. With regard to demographics, 83% of the physicians were men, and 82% were white. Their average age was 42.6 years. As for professional credentials, 42% were emergency medicine residency trained, and 58% were board certified in emergency medicine; 50% were certified by the American Board of Emergency Medicine. CONCLUSION: Given that there are 5,064 hospitals with EDs and given that the data indicate that there are 5.35 FTEs per ED, the total number of FTEs is projected to be 27,067 (SE=500). Given further that the data indicate a physician/FTE ratio of 1.47:1, we conclude that there are 39,746 persons (SE=806) needed to staff those FTEs. When adjusted for persons working at more than one ED, that number is reduced to 31,797. When the 1999 data are compared with those collected in 1997, we note a statistically significant decline in the number of hospital EDs, from 5,126 in 1997 to 5,064 in 1999 (P =.02). The total number of emergency physicians remained the same over the 2-year period, whereas the number of FTEs per institution increased from 5.11 to 5.35. The physician/FTE ratio remained unchanged. PMID- 12085067 TI - The emergency medicine workforce study: more questions than answers. PMID- 12085069 TI - Hair apposition for scalp lacerations. PMID- 12085068 TI - A randomized controlled trial comparing the hair apposition technique with tissue glue to standard suturing in scalp lacerations (HAT study). AB - STUDY OBJECTIVE: We evaluate a new technique of treating scalp lacerations, the hair apposition technique (HAT). After standard cleaning procedures, hair on both sides of a laceration is apposed with a single twist. This is then held with tissue adhesives. HAT was compared with standard suturing in a multicenter, randomized, prospective trial. METHODS: All linear lacerations of the scalp less than 10 cm long were included. Severely contaminated wounds, actively bleeding wounds, patients with hair strand length less than 3 cm, and hemodynamically unstable patients were excluded. Patients were randomized to receive either HAT or standard suturing, and the time to complete the wound repair was measured. All wounds were evaluated 7 days later in a nonblinded manner for satisfactory wound healing, scarring, and complications. RESULTS: There were 96 and 93 patients in the study and control groups, respectively. Wound healing trended toward being judged more satisfactory in the HAT group than standard suturing (100% versus 95.7%; P =.057; effect size 4.3%; 95% confidence interval 0.1% to 8.5%). Patients who underwent HAT had less scarring (6.3% versus 20.4%; P =.005), fewer overall complications (7.3% versus 21.5%; P =.005), significantly lower pain scores (median 2 versus 4; P <.001), and shorter procedure times (median 5 versus 15 minutes; P <.001). There was a trend toward less wound breakdown in the HAT group (0% versus 4.3%; P =.057). When patients were asked whether they were willing to have HAT performed in the future, 84% responded yes, 1% responded no, and 15% were unsure. CONCLUSION: HAT is equally acceptable and perhaps superior to standard suturing for closing suitable scalp lacerations. Advantages include fewer complications, a shorter procedure time, less pain, no need for shaving or removal of stitches, similar or superior wound healing, and high patient acceptance. HAT has become our technique of choice for suitable scalp lacerations.[Ong Eng Hock M, Ooi SBS, Saw SM, Lim SH. A randomized controlled trial comparing the hair apposition technique with tissue glue to standard suturing in scalp lacerations (HAT study). PMID- 12085070 TI - Bimanual laryngoscopy: a videographic study of external laryngeal manipulation by novice intubators. AB - STUDY OBJECTIVE: We perform a videographic analysis of external laryngeal manipulation (ELM) by novice intubators using a direct laryngoscopic imaging system and a validated means of assessing laryngeal view (percentage of glottic opening [POGO] score). METHODS: Nine first-year emergency medicine residents performed a total of 484 laryngoscopies during a 1-year study period, of which 271 were videotaped. Of this convenience sample, cases were included in the study if the initial laryngeal view had a POGO score of less than 50% and the quality of video imaging permitted POGO scoring before and after application of ELM. RESULTS: Seventy-two cases met the study criteria. Initial POGO scores equaled 0% in 30 cases, less than 20% in 20 cases, and 20% to 50% in 22 cases. ELM improved the POGO scores in all cases (95% confidence interval [CI] 96% to 100%) by an average of 57% (SD 23.7; 95% CI 40% to 75%). Among those with an initial POGO score equal to 0%, the mean improvement was 61% (SD 36.3; 95% CI 48% to 74%). For those with an initial POGO score of less than 20%, the mean improvement was 63% (SD 19.5; 95% CI 54% to 71%), and for those with an initial POGO score of 20% to 50%, the mean improvement was 47% (SD 16.0; 95% CI 40% to 55%). CONCLUSION: ELM is a simple and very effective technique for improving laryngeal view during laryngoscopy by novice intubators. PMID- 12085071 TI - External laryngeal manipulation: a simple intervention for difficult intubations. PMID- 12085072 TI - Teaching procedural skills to medical students: one institution's experience with an emergency procedures course. AB - STUDY OBJECTIVE: We examine the effect of a preclinical emergency procedures course on students' clinical procedural skills and medical knowledge. METHODS: This is a retrospective review of evaluation forms for a cohort of 86 students graduating from medical school at an academic center. A cross section of students (n=57) taking a clinical emergency medicine rotation over a 4-year period was also studied. Numeric scores (1 to 9 on a Likert scale) in procedural skills and medical knowledge categories were extracted from evaluations for internal medicine, surgery, obstetrics and gynecology, and emergency medicine rotations. Scores of students who had taken an elective course, Essential Procedures in Emergency Medicine (EPEM), were compared with scores of students who did not take this course. US Medical Licensing Examination Step I scores for both groups were also compared. RESULTS: Students who took EPEM scored significantly higher in the procedural skills category during the emergency medicine rotation (P =.04) and during both months of the internal medicine rotation (P =.02; P =.02). Students scored on average higher in the surgery and obstetrics and gynecology rotations, but these differences were not statistically significant. Students who took EPEM scored significantly higher in the medical knowledge category for emergency medicine (P =.01; P =.002), both months of internal medicine (P =.03; P =.006), and 1 of 2 months of surgery (P =.01) rotations. Students in obstetrics and gynecology rotations scored higher, although not significantly. US Medical Licensing Examination Step I scores were not different between students taking or not taking EPEM. CONCLUSION: Students taking EPEM achieved higher procedural skill and medical knowledge scores in clinical rotations. Emergency medicine is a specialty well suited to study procedures teaching and performance. PMID- 12085074 TI - Moonlighting and the emergency medicine resident. PMID- 12085073 TI - Improving emergency medicine residents' approach to patients with alcohol problems: a controlled educational trial. AB - STUDY OBJECTIVE: We determine whether training using a structured skills-based intervention would improve emergency medicine residents' knowledge and practice in screening and intervening with patients presenting to the emergency department with alcohol problems. METHODS: In a controlled trial conducted at 2 similar emergency medicine residency programs associated with urban, Level I trauma centers, a 4-hour didactic, video, and skills-based workshop was conducted. Main outcome measures included (1) scores on changes in self-reported knowledge, current practice, self-efficacy, role-responsibility, attitudes and beliefs, and provider readiness to change from baseline to 1 year after intervention and (2) change in practice as measured by record review before and after intervention. RESULTS: The intervention group (n=17) had a significant increase in knowledge scores (P <.001) and practice with regard to percent of medical records with evidence of screening and intervention (17% before versus 58% after; 95% confidence interval [CI] 31 to 50; P <.001); no change was observed in the control group (n=19). These increases were significantly different between groups (95% CI 30 to 54; P <.001). There were no significant differences within or between groups for composite scores derived for current practice, self-efficacy, role responsibility, or readiness to change. CONCLUSION: A brief, structured, educational intervention for residents contributed to significant improvement in knowledge and practice with regard to patients with alcohol problems. PMID- 12085075 TI - Management of corneal abrasion in children: a randomized clinical trial. AB - STUDY OBJECTIVE: We compare percentage of healing, comfort, and complications in children with corneal abrasions treated with an eye patch versus no eye patch. METHODS: We performed a randomized clinical trial of patients aged 3 to 17 years who were diagnosed with isolated corneal abrasion. Patients were randomly assigned to an eye patch or no patch group. Abrasion size was documented with digital photographs and/or an eye template diagram at presentation and at 20- to 24-hour follow-up examination. A reviewer masked to treatment group determined percent healing by measuring presentation and follow-up abrasion sizes on the photographs/template. At follow-up, interference with activities of daily living (ADL) was measured with a visual analog scale and the number of pain medication doses taken since presentation was recorded. RESULTS: A total of 37 patients were enrolled: 17 with an eye patch and 18 with no eye patch. The mean patient age was 10 years, and two thirds of the patients were male. The majority (86%) of patients had 95% or more healing at follow-up, and there was no significant difference in percent healing between the 2 groups, even when adjusted for age and initial abrasion size (95% confidence interval [CI] for the difference in means -11 to 8 and -13 to 5, respectively). There was no difference between groups for number of pain medication doses required. Among measurements of interference with ADL, only the difficulty walking score was found to be significantly different between groups (patch mean 1.7 cm [SD 2.1 cm] versus no patch mean 0.3 cm [SD 0.7 cm]; 95% CI for the difference in means 0.3 to 2.5). CONCLUSION: This study suggests that eye patching in children with corneal abrasions makes no difference in the rate of healing. There was no difference in discomfort and interference with ADL, other than greater difficulty walking in the patch group, and there were no complications in either group. PMID- 12085076 TI - A prospective study of acute, unintentional, pediatric superwarfarin ingestions managed without decontamination. AB - STUDY OBJECTIVE: We determine the incidence of clinically important bleeding in children with superwarfarin rodenticide ingestions not treated with gastrointestinal decontamination or prophylactic vitamin K. METHODS: We prospectively studied patients younger than 6 years of age who reported to our poison center with acute unintentional superwarfarin ingestions. Patients who received gastrointestinal decontamination or prophylactic vitamin K were excluded. Forty-eight- to 96-hour prothrombin time or international normalized ratio (INR) blood tests were recommended, and telephone contact was attempted at least 3 days after ingestion. RESULTS: A total of 595 consecutive patients were enrolled during the 16-month study period. Fifty patients were excluded: 8 who were known to have ingested 1 pellet or less; 25 who received activated charcoal; 15 who were treated with induced emesis; and 2 who received prophylactic vitamin K. The resulting study group contained 545 patients. Eighty-two patients were lost to follow-up. Follow-up was obtained for 463 patients, including 222 by telephone contact alone, 62 by 48- to 96-hour INR, and 179 by both methods. None of the patients had clinically important coagulopathy. Two patients had an INR of 1.5 or greater (1.5 and 1.8) without symptoms. Single nosebleeds were reported in another 2 patients with normal 48-hour INRs. Another child had a small amount of blood crusted in the nose with no other symptoms and no laboratory work available. One child with a normal 48-hour INR had blood-streaked stools that were thought to be caused by an anal fissure. CONCLUSION: Children with acute unintentional superwarfarin ingestions of less than 1 box may be managed without gastric decontamination or prophylactic vitamin K. Laboratory testing for coagulopathy should be reserved for cases involving clinically evident bleeding abnormalities. PMID- 12085077 TI - Emergency Medical Services Outcomes Project III (EMSOP III): the role of risk adjustment in out-of-hospital outcomes research. AB - The purpose of the Emergency Medical Services Outcomes Project (EMSOP) is to develop a foundation and framework for out-of-hospital outcomes research. Fundamental to that purpose is the identification of priority conditions, risk adjustment measures (RAMs), and outcome measures. In this third EMSOP article, we examine the topic of risk adjustment, discuss the relevance of risk adjustment for out-of-hospital outcomes research, and recommend RAMs that should be evaluated for potential use in emergency medical services (EMS) research. Risk adjustment allows better judgment about the effectiveness and quality of alternative therapies; it fosters a better comparison of potentially dissimilar groups of patients. By measuring RAMs, researchers account for an important source of variation in their studies. Core RAMs are those measures that might be necessary for out-of-hospital outcomes research involving any EMS condition. Potential core RAMs that should be evaluated for their feasibility, validity, and utility in out-of-hospital research include patient age and sex, race and ethnicity, vital signs, level of responsiveness, Glasgow Coma Scale, standardized time intervals, and EMS provider impression of the presenting condition. Potential core RAMs that could be obtained through linkage to other data sources and that should be evaluated for their feasibility, validity, and utility include principal diagnosis and patient comorbidity. We recommend that these potential core RAMs be systematically evaluated for use in risk adjustment of out-of hospital patient groups that might be used for outcomes research PMID- 12085078 TI - Knowledge and attitude assessment and education of prehospital personnel in child abuse and neglect: report of a National Blue Ribbon Panel. PMID- 12085079 TI - Hemorrhagic shock from a ruptured ectopic pregnancy in a patient with a negative urine pregnancy test result. AB - Ectopic pregnancy has been increasing in frequency over the past 2 decades. The sudden rupture of a fallopian tube caused by ectopic pregnancy can lead to hemorrhagic shock and death if not diagnosed and treated in a timely fashion. The emergency physician is often the health professional that is called on to make the diagnosis and coordinate timely and effective intervention. The first step in the diagnosis of ectopic pregnancy is demonstration of pregnancy by means of a rapidly performed and sensitive qualitative urine test for the beta-subunit of human chorionic gonadotropin (beta-hCG). A negative urine pregnancy test result will generally be used to exclude ectopic pregnancy from further consideration. The following is a report of a patient presenting to an emergency department with hypovolemic shock in conjunction with 2 negative urine beta-hCG analysis results and a quantitative serum beta-hCG level of 7 mIU/mL, a value less than the lower limit of detection for the highly sensitive qualitative urine and serum tests. This case report demonstrates the importance of further consideration of the diagnosis of ectopic pregnancy in the setting of a negative urine pregnancy test result. PMID- 12085080 TI - Interstitial pregnancy: a potential for misdiagnosis of ectopic pregnancy with emergency department ultrasonography. AB - Interstitial pregnancy is a rare and dangerous form of ectopic pregnancy that can be mistaken for a normal intrauterine pregnancy on ultrasonography, leading to catastrophic results. Increasingly, emergency physicians are using ultrasonography to diagnose intrauterine pregnancy. Emergency physicians should be aware of the potential for mistaking an interstitial pregnancy for an intrauterine pregnancy. We present a case report of an interstitial pregnancy misdiagnosed as an intrauterine pregnancy and discuss ultrasonographic and physical examination findings to help differentiate interstitial pregnancy from an intrauterine pregnancy. PMID- 12085081 TI - Evidence-based emergency medicine. Antibiotic treatment for acute bronchitis. PMID- 12085082 TI - Evidence-based emergency medicine. Hypertonic versus isotonic crystalloid for fluid resuscitation in critically ill patients. PMID- 12085083 TI - Update on emerging infections: news from the Centers for Disease Control and Prevention. Pertussis--United States, 1997-2000. PMID- 12085084 TI - If the walls could speak. PMID- 12085085 TI - Always believe. PMID- 12085086 TI - The epidermal growth factor receptor: a new target for anticancer therapy. PMID- 12085087 TI - Constitutional protection for animals. PMID- 12085088 TI - Magnetic stimulation reveals the distribution of language in a normal population. PMID- 12085089 TI - NMDA receptors lose their inhibitions. PMID- 12085090 TI - Short circuiting the circadian clock. PMID- 12085091 TI - Be caught napping: you're doing more than resting your eyes. PMID- 12085093 TI - Genetically engineered mouse models of neurodegenerative diseases. AB - Recent research has significantly advanced our understanding of the molecular mechanisms of neurodegenerative diseases, including Alzheimer's disease (AD) and motor neuron disease. Here we emphasize the use of genetically engineered mouse models that are instrumental for understanding why AD is a neuronal disease, and for validating attractive therapeutic targets. In motor neuron diseases, Cu/Zn superoxide dismutase and survival motor neuron mouse models are useful in testing disease mechanisms and therapeutic strategies for amyotrophic lateral sclerosis (ALS) and spinal motor atrophy, respectively, but the mechanisms that account for selective motor neuron loss remain uncertain. We anticipate that, in the future, therapies based on understanding disease mechanisms will be identified and tested in mouse model systems. PMID- 12085097 TI - Utilization of central disk of blastoderm and germinal crescent region for production of interspecific germline chimera between chicken and quail. AB - AIM: The production of interspecific germline chimeras between chicken and quail were attempted employing the dissociated cells derived from the blastodermal central disk (stage X) and the germinal crescent region of embryo (stage 7-8). METHODS: The central disk (CD) of the area pellucida in chicken blastoderm (stage X) and the germinal crescent region (GCR) of embryo (stage 7-8) were dispersed and injected into the subgerminal cavity of quail blastoderm (stage X). Injected eggs were incubated for 7 days or to hatching. The donor chicken DNA was detected by the polymerase chain reaction. RESULTS: In day-7 embryos, chicken DNA was detected in 5 gonads and 9 brains from 53 survived embryos received chicken CD cells, and 1 gonads and 6 brains from 27 survived embryos received chicken GCR. Chicken DNA was also detected from the semen of one adult male hatched from eggs received chicken GCR cells. CONCLUSION: CD and GCR cells as the donors showed the possibility to produce the interspecific germline chimera, but further studies are needed to make necessary improvement. PMID- 12085098 TI - Relationship between bicarbonate and cyclic nucleotide in the promoting effects on head-to-head agglutination in boar spermatozoa. AB - AIM: To clarify the relationship between bicarbonate and cAMP in the promoting effects on the sperm agglutination. METHODS: Spermatozoa were collected from mature boars, washed and resuspended in a modified Krebs-Ringer HEPES lacking calcium chloride (mKRH). The sperm suspensions were incubated in a water bath (38.5 degrees C) for 60 min and then the percentage of head-to-head agglutinated spermatozoa was determined. RESULTS: Supplementation of the mKRH with sodium bicarbonate (5-10 mM) significantly raised the percentage of head-to-head agglutinated spermatozoa in the samples. The addition of selective inhibitors for calcium/calmodulin-dependent phosphodiesterases (type 1: 8-methoxymethyl-IBMX and vinpocetine, 25-50 micro M) or for cAMP-specific phosphodiesterases (type 4: Ro20 1724 and rolipram, 25-50 microM) enhanced the effect of bicarbonate on sperm agglutination as highly as did the addition of non-selective inhibitors for phosphodiesterases (IBMX and papaverine, 25-50 microM). A calmodulin antagonist (W-7, 2 microM), that potentially blocks the stimulator of the calcium/calmodulin dependent phosphodiesterases, significantly enhanced the effect of bicarbonate on sperm agglutination. Moreover, a phosphodiesterase-resistant cAMP analogue (cBiMPS, 0.1 mM) markedly induced agglutination in more spermatozoa (76%) after the incubation without bicarbonate and phosphodiesterase inhibitors than did a less potent cAMP analogue (dibutyryl cAMP, 1 mM) (21%), while three kinds of cGMP analogues (0.1-1 mM) had no effect on sperm agglutination. In addition, a cAMP antagonist (Rp-cAMPS, 1 mM) significantly reduced the sperm agglutination resulting from the actions of bicarbonate and IBMX. On the other hand, the effect of bicarbonate was abolished by a change of incubation temperature from 38.5 degrees C to 25 degrees C. CONCLUSION: These findings demonstrate that the bicarbonate-induced agglutination of boar spermatozoa is controlled via the cAMP mediated, temperature-dependent signaling cascade. This cascade is suppressed by the action of the phosphodiesterase (at least types 1 and 4). PMID- 12085099 TI - Relationship between sperm mitochondrial membrane potential, sperm motility, and fertility potential. AB - AIM: To analyze the relationship between sperm mitochondrial membrane potential and sperm motility parameters by means of a computer-assisted sperm analyzer (CASA) and in-vitro fertilization rate(%FR). METHODS: Semen samples were obtained from 26 men undergoing in vitro fertilization-embryo transfer (IVF-ET). Informed consent was obtained from all men prior to the study. Samples were prepared using wash and swim-up method in HEPES-HTF medium. The sperm motility (%MOT), progressive motility (%PMOT), average path velocity (VAP) microm/s), straight line velocity (VSL) (micro m/s), curvilinear velocity (VCL) (microm/s) and %hyperactivated sperm (%HA), and the %FR were assessed. The samples were incubated in the presence of 2.0 mciromol/L of 5,5',6,6'-tetra-chloro-1,1',3,3' tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) for 30 min at 37 degrees C in air and washed in PBS before flow cytometry (FACSCalibur: Becton Dickinson) analysis. The mitochondrial probe JC-1 was used to identify the mitochondrial membrane potential. The sperm was divided into three populations according to the fluorescence pattern as follows: the high mitochondrial membrane potential group (n=8), the moderate group (n=5), and the low group (n=13). Statistical analysis was performed using unpaired t-test. RESULTS: Significant differences were found between the high and the low groups in %MOT (91.1+/-8.5 vs 63.0+/-32.7, mean+/ SD), VAP (73.0+/-14.2 vs 52.1+/-12.5), VCL (127.0+/-28.1 vs 87.0+/-22.6), %HA (27.3+/-23.6 vs 7.2+/-9.0) and %FR [73.2 (48/56) vs 59.0 (69/117)]. No significant differences were found in other CASA parameters. CONCLUSION: When the sperm mitochondrial membrane potential increases, sperm motility parameters and fertility potential will also increase. The JC-1 dye method is useful to predict sperm fertility potential. PMID- 12085100 TI - Distribution of intracellular and extracellular expression of transforming growth factor-beta1 (TGF-beta1) in human testis and their association with spermatogenesis. AB - AIM: Spermatogenic dysfunction may result from thickening of seminiferous tubular basement membrane (BM) with tubular sclerosis. Transforming growth factor beta1 (TGF-beta1) plays an important role in fibrogenesis. The intracellular and extracellular expression of TGF-beta1 in the testis were immunohistochemically determined, using LC antibody (LC) for intracellular TGF-beta1 and CC antibody (CC) for extracellular TGF-beta1. METHODS: Twenty-three testicular biopsy specimens were obtained from varicocele and five from Sertoli-cell-only (SCO) patients, and five from normal volunteers. The relative area involved by the expression of TGF-beta1 for CC or LC (TGF-beta1 index for CC or LC) was examined, and semen parameters and serum hormonal levels and TGF-beta1 were analyzed. The Johnson score (JS), the BM thickness, and the tubular diameter were also determined. RESULTS: Immunoreactivity for CC was hardly detected. That for LC was detected in the Sertoli and germ cells. The TGF-beta1 index for LC was significantly higher in the varicoceles than in the normal testes. Interestingly, that for LC was significantly higher in the varicoceles than in the SCO. The level of serum TGF-beta1 was significantly higher in varicoceles than in the normal testes. CONCLUSION: The distribution of the intracellular and extracellular expression of TGF-beta1 in human testis was demonstrated. It suggests that TGF-beta1 is related to fibrosis of seminiferous tubules and may lead to spermatogenic disruption. PMID- 12085102 TI - Bone mineral density in hypogonadal men remains low after long-term testosterone replacement. AB - AIM: In 11 congenital hypogonadal men, the bone mineral density (BMD) values were determined to assess the effect of long-term androgen replacement therapy (ART) on skeletal integrity. METHODS: Eleven congenital hypogonadal men, including 8 isolated gonadotropin deficiency patients, 2 Kallmann's syndrome and 1 vanishing testes syndrome were recruited and treated with 250 mg of testosterone enanthate intramuscularly every 4 weeks for 7-43 years (mean+/-SD: 21.5 +/-13 years). In these patients and a group of 10 healthy young men (controls), the whole and trabecular BMDs were examined at the distal end of radius by means of a peripheral quantitative computerized tomography device. RESULTS: The whole radial BMD in hypogonadal men was significantly less in the patients than in the healthy men (498+/-115 and 725+/-134 mg/cm(3), respectively; P<0.01); the trabecular BMD was also lower in the hypogonadal men (199+/-80 and 375+/-89 mg/cm(3); P< 0.01). The whole radial BMD values in 10 of 11 hypogonadal men were at least 1 SD below the mean value for healthy young men; 2 hypogonadal men had BMD values more than 2.5 SD lower than the healthy mean. Additionally, the whole radial BMD showed a significant negative correlation with the patient's age at the initiation of ART (r = 0.748, P<0.01). The serum level of bone-specific alkaline phosphatase and the urinary level of deoxypyridinoline were not significantly different between the two groups. CONCLUSION: Osteopenia persists in the hypogonadal men after long term ART, suggesting that such patients have a persistent defect in bone development not alleviated by androgen replacement. PMID- 12085101 TI - Microdeletions in the Y chromosome of patients with idiopathic azoospermia. AB - AIM: To evaluate the occurrence and prevalence of microdeletions in the gamma chromosome of patients with azoospermia. METHODS: DNA from 29 men with idiopathic azoospermia was screened by polymerase chain reaction (PCR) analysis with a set of gamma chromosome specific sequence-tagged sites (STSs) to determine microdeletions in the gamma chromosome. RESULTS: Deletions in the DAZ (deleted in azoospermia) loci sgamma254 and sgamma255 were found in three patients with idiopathic azoospermia, resulting in an estimated frequency of deletions of 10.7% in idiopathic azoospermia men. CONCLUSION: We conclude that PCR analysis is useful for the diagnosis of microdeletions in the Y chromosome, which is important when deciding the suitability of a patient for assisted reproductive technology such as testicular sperm extracion-intracytoplasmic sperm injection (TESE-ICSI). PMID- 12085103 TI - Mechanisms of metastasis suppression by introduction of human chromosome 10 into rat prostate cancer. AB - AIM: The metastatic ability of a Dunning R-3327 rat prostate cancer subline (AT6.3) was suppressed by the introduction of human chromosome 10, when these hybrid cancer cells were injected subcutaneously into nude mice (Nihei et al., Genes Chromosomes Cancer 14:112-119, 1995). The present study was undertaken to clarify which step of metastasis was suppressed in the human chromosome 10 containing microcell hybrids (AT 6.3-10 clones). METHODS: Gelatin zymography, an in vitro invasion assay using a Boyden chamber and an intravenous metastasis assay involving the injection of hybrid cells into nude mice were performed. RESULTS: Gelatin zymography revealed that AT6.3-10 microcell hybrid clones expressed the 72 kD type IV collagenase (MMP-2) at an almost equal level as control microcell hybrid clones. Both the invasiveness as measured by the invasion assay and the number of lung metastases as measured by the intravenous metastasis assay of AT6.3-10 hybrid clones were significantly less than those of the AT6.3 parental clone. CONCLUSION: The human chromosome 10 suppresses both the local invasion and the metastatic process after entry into the blood circulation of rat prostate cancer. This decrease in local-invasive ability does not seem to require a decrease in MMP-2 activity. PMID- 12085104 TI - Suppression of metastasis of rat prostate cancer by introduction of human chromosome 13. AB - AIM: Chromosome 13 is one of the most frequently altered chromosomes in prostate cancer. The present study was undertaken to examine the role of human chromosome 13 in the progression of prostate cancer. METHODS: Human chromosome 13 was introduced into highly metastatic rat prostate cancer cells via microcell mediated chromosome transfer. RESULTS: Microcell hybrid clones containing human chromosome 13 showed suppression of metastasis to the lung without any suppression of tumorigenicity, except for one clone, which contained the smallest sized human chromosome 13 and did not show any suppression on lung metastasis. Expression of two known tumor suppressor genes, BRCA2 and RB1, which map to chromosome 13, was examined by reverse transcription- polymerase chain reaction analysis. BRCA2 was expressed only in the metastasis-suppressed microcell-hybrid clones, whereas RB1 was expressed in all clones. CONCLUSION: Human chromosome 13 contains metastasis suppressor gene(s) for prostate cancer derived from rat. Furthermore, the RB1 gene is unlikely to be involved in the suppression of metastasis evident in this system. PMID- 12085105 TI - The influence of cryoprotective media and processing procedures on motility and migration of frozen-thawed human sperm. AB - AIM: The study was designed to examine the effects of cryoprotective media, and glycerolating and thawing procedures on human sperm motility and gel penetrating ability. METHOD: Fifteen unselected donors provided semen varying in quality that was distributed in a factorial design across three cryoprotectants (glycerol, egg yolk-citrate-glucose-glycerol and egg yolk-tris-glucose-glycerol). Also, glycerol was added at room temperature versus at 4 degrees C. Two thaw temperatures were tested (laboratory air temperature for 10 min versus a 65 degrees C waterbath for 4 seconds). The proportion of total and progressively motile sperm was estimated immediately after thawing and following incubation at 35 degrees C for 2 h. Migration of sperm for 30 min at 37 degrees C through polyacrylamide gel was tested. RESULTS: Donors differed greatly, with post-thaw total motility of sperm ranging from 9 to 44% (P<0.05). Egg yolk-citrate-glucose-glycerol and egg yolk tris-glucose-glycerol were superior to glycerol alone (post-thaw values of 35, 37 and 21%, respectively, P<0.05). This was due primarily to poor sperm survival when semen was cooled to 4 degrees C without glycerol or egg yolk. The two thaw temperatures gave similar results. Sperm migration tests paralleled the motility results, but were more sensitive in detecting differences. CONCLUSION: Egg yolk, particularly in a tris-based medium that is widely used in domestic animals, improved the cryopreservation of both good and poor quality human semen. PMID- 12085106 TI - Testicular dysfunction in BALB C mice with Schistosoma intercalatum bilharziasis. AB - AIM: To evaluate the effect of Schistosoma intercalatum infestation on the testicular function of mice. METHODS: Male BALB C mice were infested by immersion of the tail and hind feet into the water with 50 or 100 cercariae of Cameroon strain S. intercalatum. Sixty days later the animals were killed, blood was collected and the testis, epididymis and seminal vesicles were dissected and weighed. The plasma and testicular testosterone were evaluated with radioimmunoassay, the seminal vesicular fructose with colorimetric method, and the histology of testis and cauda epididymis observed under light microscope. The intensity of infestation was estimated in terms of S. intercalatum egg load in the liver. RESULTS: In infested mice, the testicular weight did not change significantly while the epididymal and seminal vesicular weights were significantly lowered compared to the controls. Furthermore, the fructose levels in the seminal vesicle fluid were significantly (P<0.01) reduced in about 50% of infested mice. S. intercalatum infestation also decreased the plasma and testicular testosterone concentrations. Histological studies indicated that the spermatogenesis, the testicular interstitial tissue and the cauda epididymis were qualitatively normal. Parasite eggs were not found in these organs. The mean seminiferous tubular diameter did not show significant differences between the infested and control mice. CONCLUSION: S. intercalatum infestation impairs testicular function. PMID- 12085107 TI - Cryptorchidism and AZF microdeletion. PMID- 12085108 TI - Antioxidative effect of fullerenol on goat epididymal spermatozoa. AB - AIM: To evaluate the effect of fullerenol on the antioxidant system of goat epididymal sperm. METHODS: Fresh epididymides of adult goats were obtained from local slaughter houses and sperm were collected by chopping the epididymis in modified Ringer's phosphate solution (RPS medium). After several washings the sperm samples were equally dispersed in RPS medium and incubated with fullerenol (1, 10 and 100 micromol) and FeSO(4)/ascorbate (40/200 micromol) with or without fullerenol (1, 10 and 100 micromol) for 3 h at 32 degree C. After incubation, an aliquot of sperm samples were homogenized and centrifuged and the supernatant used for biochemical studies. RESULTS: In FeSO(4)/ascorbate-incubated samples, the activities of antioxidant enzymes, superoxide dismutase, glutathione peroxidase and glutathione reductase, were decreased while lipid peroxidation increased as compared to the control sperm samples. In fullerenol-incubated sperm samples, the activities of superoxide dismutase, glutathione peroxidase and glutathione reductase were increased while lipid peroxidation was decreased in a dose-dependent manner. Co-incubation of sperm with fullerenol (1,10 and 100 micromol) and FeSO(4)/ascorbate (40/200 micromol) increased the activities of antioxidant enzymes and prevented the iron-induced elevation of lipid peroxidation in a dose-dependent manner. CONCLUSION: Fullerenol reduces iron induced oxidative stress in epididymal sperm of goat by increasing the activities of antioxidant enzymes and decreasing lipid peroxidation. PMID- 12085109 TI - Cr(V) involvement in the toxicity pathway of testicular damage. AB - AIM: The functional integrity of the blood-testis barrier (BTB) in male mice exposed to Cr(V) was studied in order to clarify the mechanism underlying testicular injury. METHODS: Adult male mice were subcutaneously injected repeated doses of 8.02 micromol (0.5 ml) of Cr/mouse.day for 5 days. Animals receiving a similar volume of bis(hydroxyethyl)-aminotris(hydroxymethyl)methane buffer (BT) were used as controls. The animals were sacrificed on day 6 and small fragments of seminiferous tubules, approximately 8-10 mm length, were incised and sutured at both ends. They were exposed in vitro to horseradish peroxidase-containing culture medium for 10 minutes. Tissues were then fixed and processed for ultrastructural studies. RESULTS: Controls and Cr(V)-treated group resulted in the uptake of the tracer by Sertoli cells. However, the major finding consisted in the permeability of the BTB only in the Cr(V)-group, as evidenced by the presence of the tracer within the junctions between the neighbouring Sertoli cells. CONCLUSION: The BTB is disrupted in mice submitted to Cr(V). The permeability of the BTB is a crucial feature to be investigated for the understanding of lesions within the seminiferous tubule. PMID- 12085110 TI - Luteinizing hormone pulse frequency and amplitude in azoospermic, oligozoospermic and normal fertile men in Turkey. AB - AIM: To investigate the LH pulse frequency and amplitude in azoospermic and oligozoospermic patients and to compare them with normal fertile subjects. METHODS: In this controlled clinical study, 10 normal fertile male volunteers and 20 infertile patients (10 oligozoospermic and 10 azoospermic) were enrolled. Blood samples were taken every 30 minutes for 12 hours. FSH, LH and T levels were determined. LH was observed at all the blood samples, but FSH and testosterone only at the first, middle and last samples. RESULTS: The mean LH levels were significantly different between all the groups, but there was no statistical difference in the FSH levels between the fertile and oligozoospermic groups. The mean LH levels increased from the fertile towards the azoospermic groups (P<0.01). The LH pulse amplitude and frequency were significantly different between all the 3 groups. The former increased while the latter decreased from the fertile to the azoospermic groups. The T levels were different statistically only between the fertile and the azoospermic groups. CONCLUSION: The more prominent is the testicular defect, the lower will be the LH pulse frequency and the higher the amplitude. PMID- 12085123 TI - Powered phlebectomy (TriVex) in treatment of varicose veins. AB - This study assesses the operation of transilluminated powered phlebectomy for removal of varicose veins. It was a prospective, noncomparative, multicenter, pilot study designed to evaluate the safety and efficacy of the powered varicose vein extractor for ablation of primary varicose veins. A total of 114 patients (117 limbs) were recruited from four centers in Europe and four centers in the United States. Safety of the varicose vein extractor was evaluated by recording nature and severity of all adverse events and complications. Efficacy was assessed by the patient, an independent study nurse, and the surgeon. Operations were performed under general, spinal, or epidural anesthesia and tumescent anesthesia was added with infusions of dilute lidocaine with epinephrine. Transillumination was achieved with a specially designed cannula, and the vein extraction was done using a vein resector with a rotating tubular inner cannula encased in a stationary outer sheath dissector. Demographic information regarding the 28 men and 89 women included in the study are detailed. Eighty-four percent of the limbs were CEAP class 2 with only 16% being in classes 3 and 4. Accompanying greater saphenous vein stripping was done in 67% of the limbs in the United States and 88% in those in Europe. Proximal ligation only was used in one limb in the United States and eight in Europe. The study showed that transilluminated powered phlebectomy used in varicose vein removal is swift and efficacious with a conservation of operating time and the results being satisfactory to the patient and clinician alike. PMID- 12085124 TI - Incidental discovery of an inferior vena cava aneurysm. AB - Inferior vena cava (IVC) aneurysms are extremely rare, with only 18 reported cases in the world literature. These aneurysms are categorized as acquired, congenital, or associated with arteriovenous fistulae. Thrombosis of an IVC aneurysm can lead to IVC syndrome characterized by massive lower extremity edema, pulmonary embolism, or even death. Therapeutic alternatives range from watchful waiting to operative resection. This report presents a case of an IVC aneurysm noted incidentally at the time of diagnostic computed tomography for the evaluation of blunt chest trauma following a motor vehicle collision. In addition, the classification, embryology, diagnosis, and management of this unusual clinical entity are reviewed. PMID- 12085125 TI - Erythrocyte mean corpuscular volume associated with severity of peripheral arterial disease: an angiographic evaluation. AB - Elevated erythrocyte mean corpuscular volume (MCV) may be a risk factor for peripheral arterial disease (PAD). The aim of the present study was to evaluate whether MCV was associated with the severity of atherosclerotic findings in the lower limbs of PAD patients, as measured by an angiographic scoring system based on vessel lumen reduction. One hundred male patients with symptomatic PAD were studied. MCV was significantly correlated with the angiographic score (rs = 0.247, p = 0.013). PAD patients with an angiographic score in the lower third were compared to those with values in the upper third using a logistic regression model with age, smoking, hypertension, MCV, homocysteine, and total cholesterol and triglycerides as independent variables. This model revealed significant odds ratios (OR) for MCV (OR = 2.02 for an increment of 5 fl, 95% CI = 1.08-3.8) and for age (OR = 2.41 for an increment of 10 years, 95% CI = 1.21-4.81) and facilitated classification of 71% of all subjects correctly. In conclusion, MCV may be associated with angiographically determined disease severity in patients with PAD. This finding supports the hypothesis that MCV is a risk factor for PAD, although the mechanism by which MCV may contribute to the presence and severity of the disease is not yet determined. PMID- 12085126 TI - The impact of peripheral and coronary artery disease on health-related quality of life. AB - Vascular diseases, like peripheral arterial diseases (PAD) and coronary artery disease (CAD), are common diseases with a high morbidity. We investigated and compared the impact of these diseases on daily life using health-related quality of life (HRQOL) analysis. In 89 patients with PAD and 89 patients with multivessel CAD, the SF-36 was assessed before diagnostic staging and compared with an age-matched reference population. Both groups of patients had a significantly impaired HRQOL as compared to a reference population. Patients with PAD scored significantly (p < 0.001) lower than those with CAD on the domains general health perception, change in health, physical functioning, social functioning, and pain. Patients with PAD have a reduced HRQOL in comparison with patients with CAD. Each disease has an impact on a specific domain of the HRQOL. Vascular disease mainly influences physical health. PMID- 12085127 TI - Traumatic bilateral ECCA injury in a roller coaster enthusiast. AB - Vascular and trauma surgeons have seen a marked increase in the incidence of traumatic injury of the ECCA. Making the diagnosis is straightforward, but requires a high index of suspicion. This patient's injury is from hyperextension/flexion trauma that occurred from repetitive rides on roller coasters. PMID- 12085128 TI - Urgent carotid artery repair: retrospective study of 21 cases. AB - The purpose of this retrospective study was to assess the outcome of urgent surgical repair in patients with symptomatic carotid artery lesions. The study population included 21 patients who underwent carotid artery repair less than 24 hr after diagnostic work-up. Indications included crescendo transient ischemic attacks in 5 cases, fluctuating neurological deficits in 11, and stroke in evolution in 5. Immediate mortality was 9.5%. Mean follow-up was 55 months. Actuarial survival rates at 1 and 5 years were 90% and 62%, respectively. Actuarial neurological deficit-free rates at 1 and 5 years were 95% and 76%, respectively. The results of this study document the effectiveness of urgent carotid artery repair for treatment of acute cerebral ischemia. Although higher than elective surgery, morbidity and mortality are acceptable, given the severity of illness in this patient population. PMID- 12085130 TI - What's right when the gallbladder's left? A case report. AB - During laparoscopic cholecystectomy for cholelithiasis in a 40-year-old woman, we came upon a case of incidentally discovered left-sided gallbladder (LSG). Two anatomic variants of LSG are known: (a) "true LSG," in which, according to Gross, an accessory gallbladder originates from the left hepatic duct (LHD), the right embryonic bud is readsorbed, and the cystic duct joins either the CBD from the left or the LHD directly. Otherwise, a normal right-sided gallbladder adheres to the inferior left hepatic lobe, and the cystic duct joins the CBD from the right side (as in our case); (b) gallbladder under the fourth hepatic segment, medial to a right-sided round ligament, probably resulting from a prenatal obliteration of the right umbilical vein. Left-sided gallbladder is a paraphysiologic condition that when identified before surgery, must be studied by CT or MRI, when incidentally discovered during surgery must be promptly recognized by the surgeon, who must be aware of the unpredictable confluence of the cystic duct into the CBD. The following operative precautions are useful for avoiding a lesion of the CBD: The surgeon should start dissection of Calot's triangle as close as possible to the gallbladder margin, prepare and clip the cystic duct as close as possible to the infundibulum, and a 30 degrees angled telescope. If in doubt, the surgeon should perform an intraoperative cholangiography. PMID- 12085131 TI - Thoracoscopic-assisted management of postpneumonic empyema in children refractory to medical response. AB - BACKGROUND: Empyema frequently complicates the hospitalization of children; and in advanced stages, it often requires surgical intervention. In this study, we investigated the use of video-assisted thoracic surgery (VATS) for the management of postpneumonic empyema in children who have had an unsatisfactory medical response. METHODS: We did a retrospective review of the medical records of 51 consecutive patients with loculated empyema (mean age, 5 years; range, 2 months to 15 years) hospitalized at Chang Gung Memorial Hospital between 1995 and 2000. All patients underwent debridement of the necrotic lung tissue and evacuation of the loculated empyema cavity using a VATS approach. RESULTS: The mean operating time for the 51 patients was 90 min (range, 50-210); mean blood loss was 70 cc. Fever subsided within 72 h postoperatively in all patients. On average, chest tubing was removed on the 7th postoperative day (range, 4-18 days). However, in one patient who suffered from a prolonged air leak, the chest tube was not removed until day 18. The mean postoperative stay for all patients was 13.7 days (range, 9-23). No deaths occurred, and all of the children made a good recovery. A follow-up revealed that one of the 51 children patient suffered a left upper lung abscess 7 months after discharge. Left upper lobectomy was performed in this case, and the patient was discharged uneventfully 10 days after the operation. CONCLUSIONS: VATS is a safe and effective treatment for pediatric empyema. Thoracoscopic-assisted surgery facilitates visualization, evacuation, and debridement of the necrotizing lung tissue. Early surgical intervention can avoid lengthy hospitalization and prolonged intravenous antibiotic therapy, and it can accelerate clinical recovery. PMID- 12085133 TI - Ergonomic aspects of four different types of laparoscopic instrument handles with respect to elbow angle. An electromyogram-based study. AB - BACKGROUND: Only few studies have tested different ergonomic aspects of the working posture assumed by laparoscopic surgeons. Although no experimental data are available for a laparoscopic setting, a working posture with a horizontal forearm or an elbow angle 90 degrees to 120 degrees has been recommended for performing minimally invasive surgery (MIS). The comparison of electromyogram (EMG) activities in different muscles provides information about the force developed by each muscle and allows assessment of its contribution to a functional movement. The current study aimed to investigate whether certain handles do not support this posture. METHODS: For this study, 12 volunteers were postured in two different standardized arm positions, defined by elbow angles of 90 degrees and 120 degrees. They were manipulating a 0.1-N and a 2.5-N microswitch with four different types of instrument handle design: axial handle, ring handle, shank handle, Hirschberg handle. During the test, the EMG activities of five forearm muscles were recorded and normalized with respect to the maximum voluntary activity of the respective muscle. RESULTS: Virtually no significant difference in EMG activity was found between the two elbow angles in any of five forearm muscles for a simple grasping maneuver. Thus, the muscle activity required to manipulate different types of MIS handles is similar for the elbow angles of 120 degrees and 90 degrees. CONCLUSIONS: The current study did not show relevant differences between the two elbow angles for any of the four handles during a simple grasping maneuver with respect to the force required in the main forearm muscles. PMID- 12085134 TI - Laparoscopic resection of perforated Meckel's diverticulum in a patient with clinical symptoms of acute appendicitis. AB - We report the laparoscopic resection of a perforated Meckel's diverticulum (MD) found in a 14-year-old boy who presented with abdominal pain and nausea. There was rebound tenderness in the right lower quadrant of the abdomen, which appeared suspicious for acute appendicitis. The patient was referred to the operating room, and laparoscopic appendectomy was performed. With the appendix showing no macroscopic signs of inflammation, laparoscopy was continued and a perforated MD was identified 50 cm proximal to the ileocecal valve. The findings included pus and localized peritonitis between the ileal loops adjacent to the perforation site. The diverticulum was longitudinally resected with an Endo-Gia stapler. The histopathologic workup confirmed the diagnosis of a perforated MD. The patient completely recovered and was discharged 8 days after the procedures. At this writing, he is completely asymptomatic 6 months later. We conclude from our observation that laparoscopic resection of a perforated MD can be performed safely even when localized peritonitis is present. Inspection of the small intestine should be performed to exclude a symptomatic or perforated MD when the appendix does not show any signs of acute appendicitis. PMID- 12085135 TI - Laparoscopic resection of patent omphalomesenteric duct in an adult. AB - The omphalomesenteric duct is the embryonic structure connecting the yolk sac to the primitive gut that disappears at 8 to 9 weeks of gestation. Failure of this duct to involute results in various anomalies. Failure of complete closure at different stages results in various anomalies, which occur in approximately 2% of the population. However, complete patency of omphalomesenteric duct is rare, reportedly occurring in approximately 15% omphalomesenteric duct anomalies. Adult presentations are extremely rare. We report the case of a 44-year-old man with a patent omphalomesenteric duct (POMD). He was admitted with a foul-smelling discharge from the umbilicus that he had experienced for 5 years. The condition had been interpreted as an umbilical granuloma at other centers, and multiple unsuccessful sessions of cauterization with silver-nitrate bar had been performed. At laparoscopy through three ports, the POMD was resected with the aid of a laparoscopic linear stapler. To our knowledge, this is the first adult case of POMD resected laparoscopically. The pitfalls of this diagnosis in the adult and the technique of laparoscopic resection is discussed in light of the available literature. PMID- 12085136 TI - Pseudomonas aeruginosa liver abscesses after diagnostic endoscopic retrograde cholangiography in two patients with sphincter of Oddi dysfunction type 2. AB - Patients with sphincter of Oddi dysfunction have a significantly increased rate of pancreatitis after manometry or sphincterotomy, but septic complications after diagnostic endoscopic retrograde cholangiopancreatography (ERCP) in patients with sphincter of Oddi dysfunction type 2 have not been reported. We describe two patients with sphincter of Oddi dysfunction type 2 in whom Pseudomonas aeruginosa serotype 10 septicemia and multiple small hepatic abscesses developed, all within 48 h after they underwent diagnostic ERCP. The sepsis and hepatic abscesses resolved after successful intravenous antibiotic administration. Despite scrupulous examination of the duodenoscope washing machine and the bottle of water, the bacteria responsible for the sepsis could not be isolated. It is possible that despite disinfection, a nondetectable colony of P. aeruginosa remained in a part of duodenoscope and proliferated to reach a potentially hazardous level the following day. This report highlights the importance administering antibiotic prophylaxis to patients with sphincter Oddi dysfunction type 2 who undergo ERCP, despite the functional nature of the disease. PMID- 12085137 TI - Hand-assisted laparoscopic anatomical left lobectomy using hemihepatic vascular control technique. AB - We report a new approach for laparoscopic anatomical left lobectomy. Although laparoscopic limited resection of the liver has been reported, major liver surgery with a laparoscopic approach remains uncommon. Obstacles to routine laparoscopic surgery on the liver are mainly related to difficulty in retraction with current instrumentation, difficulty in assessing safe margins of resection without the use of tactile sense, and the difficulty of safe parenchymal dissection laparoscopically. We introduce a hand-assisted method that can help in resolving the difficulties and pitfalls associated with laparoscopic liver resection, and in making this surgery safer. The hand is the best atraumatic liver retractor in laparoscopic resection and facilitates the use of laparoscopic ultrasonography. Stable hemostasis can be achieved by proper manual application of vascular clips in case of vascular injury. The hemihepatic inflow control technique used in the present case was the en masse occlusion of Glisson's sheath of the left hemipedicle at the bifurcation. This technique was used exactly the same as in open surgery. Major vessels such as the left hemipedicle and left hepatic vein were dissected by endovascular cutter. The patient had an uneventful, quick postoperative recovery. This technique allows a minimally invasive anatomical major surgery for liver tumors. PMID- 12085138 TI - Gallstone ileus as a complication of acute cholecystitis. Laparoscopic diagnosis and treatment. AB - Gallstone ileus accounts for 1-4% of all cases of intestinal obstruction, with its incidence rising with age of patients. There is often a long delay between onset of symptoms (usually abdominal pain, vomiting, and bowel distension) and proper treatment, with a simple enterolithotomy as the one of choice. We report a case of an atypical gallstone ileus presented as a complication of acute cholecystitis, treated with a laparoscopic guided enterolithotomy. A 67-year-old woman on the 5th p.o. day after a laparoscopic procedure for an empyematous cholecystitis (no sign of fistula or duodenal perforation and a "negative" intraoperative cholangiography) presented continuous vomiting as the only symptoms of a subileus (radiographic diagnostic images negative for intestinal obstruction or intraluminal gallstone or duodenal fistula). A laparoscopic diagnostic approach revealed a gallstone in the distal jejunum. Through a 5 cm midline incision the intestine, including the gallstone, was brought out extracorporally and the stone was removed by a simple enterolithotomy. The postoperative course was uneventful and the patient had no complaint at a 1-year follow-up. We consider the laparoscopic approach, in patients with "abdominal emergencies," feasible and safe in experienced hands. It provides diagnostic accuracy as well as therapeutic capabilities, as in the case of gallstone ileus we have reported. PMID- 12085139 TI - Comparison of interleukin-6, interleukin-8, and granulocyte colony-stimulating factor production by the peritoneum in laparoscopic and open surgery. AB - BACKGROUND: Human mesothelial cells secrete a variety of cytokines. The levels of postoperative serum inflammatory cytokines are thought to reflect the magnitude of surgical stress. METHODS: Pieces of peritoneum were obtained immediately upon and 1 h after entry into the abdominal cavity in nine patients undergoing laparoscopic surgery and 11 patients undergoing open surgery. The samples were cultured and interleukin (IL)-6, IL-8, and granylocyte colony-stimulating factor (G-CSF) levels in the supernatants were measured by enzyme-linked immunosorbent assay (ELISA). Expression of IL-6, IL-8, and G-CSF mRNAs was examined by RT-PCR. RESULTS: At 1 h after laparotomy, the amounts of IL-6 and G-CSF produced by the peritoneum were significantly greater than those obtained immediately after the procedure, but this difference was not observed with laparoscopic surgery. Reverse transcription-polymerase chain reaction (RT-PCR), which showed an increase in the expression of cytokine mRNAs at 1 h after laparotomy, was compatible with these results. CONCLUSION: The lower levels of cytokine production by the peritoneum suggest that laparoscopic surgery is associated with lower degree of surgical stress. PMID- 12085141 TI - A further modification of fundoplication. 90 degrees anterior fundoplication. AB - BACKGROUND: Laparoscopic Nissen fundoplication is the most widely applied procedure for the surgical treatment of gastroesophageal reflux. However, it can be followed by adverse outcomes, including dysphagia and "wind-related" problems. To reduce the likelihood of side effects, we have progressively modified this procedure to an anterior 90 degrees partial fundoplication. METHODS: The procedure entails posterior hiatal repair, posterior esophagopexy, accentuation of the angle of His, and construction of a 90 degrees anterior partial fundoplication. Clinical follow-up was performed prospectively using a standardized questionnaire. RESULTS: From February 1999 to June 2001, 83 patients underwent 90 degrees anterior fundoplication for gastroesophageal reflux disease. In 45 the procedure was chosen because of specific patient or surgeon preference, and in 38 it was performed within the context of an ongoing randomized trial. Operating time ranged from 20 to 140 minutes (median, 52 min), and all but one of the procedures were completed laparoscopically. One patient experienced a major postoperative complication--small bowel injury from Veress needle. Follow-up extends up to 2 years (median, 1 year). Two patients have undergone further surgery, both for recurrent reflux. Control of reflux has been acceptable, with a reduction in heartburn symptom scores and high overall satisfaction. Postoperative dysphagia measured using a visual analog scale was less following surgery compared with preoperative scores. Eighty-two percent of patients could belch normally 3 and 12 months after surgery. CONCLUSIONS: Ninety-degree anterior fundoplication achieves good control of reflux and a low incidence of side effects. To further evaluate its potential, we are currently undertaking a prospective randomized trial. PMID- 12085142 TI - Wound complications of laparoscopic vs open colectomy. AB - BACKGROUND: This study was conducted to determine if laparoscopic colon surgery has changed the incidence of wound complications after colon resection. METHODS: Eighty-three patients were randomized to undergo either laparoscopic (LCR) or open colon resection (OCR) for cancer at our institution as part of a multicenter trial. Data were tabulated from review of the prospective database and physician records. RESULTS: Thirty-seven patients were randomized to LCR and 46 to OCR. Seven patients in the LCR group were converted to OCR. LCR was performed using a limited midline incision for anastomosis and specimen extraction. Incision length was significantly greater (p <0.001) in the OCR group (19.4 +/- 5.6 cm) compared to the LCR extraction site (6.3 +/- 1.4 cm). Wound infections occurred in 13.5% of patients after LCR (2.7% trocar, 10.8% extraction sites) and in 10.9% of patients after OCR. Over a mean follow-up period of 30.1 +/- 17.8 months, incisional hernias developed in 24.3% of patients after LCR and 17.4% after OCR. In the LCR group, extraction sites accounted for 85.7% of all wound complications. CONCLUSIONS: The extraction site for LCR is associated with a high incidence of complications, comparable to open colectomy. Strategies to alter operative technique should be considered to reduce the incidence of these complications. PMID- 12085145 TI - Effectiveness of the clinical pathway to decrease length of stay and cost for laparoscopic surgery. AB - BACKGROUND: Although clinical pathways have become popular strategies to improve the quality of medication in the field of laparoscopic surgeries, their economical effectiveness is not well defined. The aim of this study was to investigate the effect of clinical pathways for laparoscopic surgeries on cost and length of hospital stay. METHODS: From January 2000 to June 2001, clinical pathways were introduced for laparoscopic surgeries, such as laparoscopic cholecystectomy (Lap. C, n = 210), laparoscopically assisted distal gastrectomy with Billroth-I reconstruction (Lap. B-I, n=33), and laparoscopically assisted colectomy (Lap. colon, n=34). We compared total lengths of hospital stay and the economical efficiency before and after pathway implementation at Wakayama Medical University Hospital. RESULTS: The length of hospital stay in Lap. C was shortened from 7.8+/-2.6 (mean+/-SD) days to 6.9+/-2.0 days (p = 0.03) and the total costs during hospitalization decreased from yen 509,320+/-58,800 to yen 489,130+/ 43,860 (p=0.009), resulting in less burden for patients. At the same time, the daily costs were increased from yen 66,230+/-8920 to yen 70,840+/-6820 (p=0.0001), indicating that more efficient medical care was being given to patients. Similar results were obtained in Lap. B-I and Lap. colon groups. CONCLUSIONS: In our study, the implementation of clinical pathways in the field of laparoscopic surgeries produced significant decreases in length of total hospital stay and cost while maintaining the quality of patient outcomes. PMID- 12085146 TI - Video-assisted thoracoscopic esophagectomy and radical lymph node dissection for esophageal cancer. A series of 75 cases. AB - BACKGROUND: The efficacy of thoracoscopic radical esophagectomy for cancer has yet to be established, mainly because previous reports have not included a sufficient number of cases. METHODS: Seventy-five treatment-naive patients with esophageal cancer without contiguous spread underwent esophageal mobilization and extensive mediastinal lymphadenectomy through a 5-cm mini-thoracotomy and four trocar ports. RESULTS: Video-assisted thoracoscopic surgery was performed without major intraoperative complications or emergency conversion to open surgery. We retrieved 34.1+/-13.0 mediastinal nodes, including 11.5+/-3.8 tracheobronchial nodes and 6.2+/-3.0 recurrent laryngeal nodes. Mean time of operation and blood loss were less in the last 39 patients than the first 36 (186.7+/-25.3 min and 165.4+/-101.8 g vs 270. 2+/-96.0 min and 421.5+/-31.2 g, respectively: p <0.0001 and p <0.001). Pulmonary morbidity was 5% in the later 39 patients. Survival was 90%, 80%, and 57% at 1, 2, and 5 years after surgery. CONCLUSION: Thoracoscopic radical esophagectomy has less morbidity and comparable survival to conventional surgery, after a moderate amount of experience. Mini-thoracotomy is essential to perform the procedure safely and effectively. PMID- 12085147 TI - Laparoscopic cholecystectomy in 226 cirrhotic patients. Experience of a single center in Taiwan. AB - BACKGROUND: Since 1987, laparoscopic cholecystectomy (LC) has been widely used as the favored treatment for gallbladder lesions. However, cirrhosis and portal hypertension have been considered relative or absolute contraindications to LC. This study aimed to assess the safety of LC in cirrhotic patients. METHOD: The medical records of 226 cirrhotic patients with gallbladder lesions who had undergone LC from 1991 to 2000 were reviewed, including demographics, severity of cirrhosis, laboratory data, operative morbidity, operative mortality, and hospital stay. Furthermore, the clinical features and outcomes of 4030 patients with gallbladder lesions without cirrhosis that had undergone LC were also summarized for comparison. RESULTS: Of 4256 patients with gallbladder lesions that had undergone LC, 226 (5.6%) had cirrhotic livers. The cirrhotic group patients clearly exhibited the trends of older age, worse liver function, higher blood loss, and higher mortality compared to the noncirrhotic group patients. CONCLUSIONS: This study presents the clinical features and outcomes of 226 cirrhotic patients who underwent LC. LC, once considered contraindicated in patients with cirrhosis, is a feasible procedure for most Child's A and B patients with cholecystolithiasis. Minor morbidity, an acceptable conversion rate, and shorter hospital stay can be achieved by applying LC to treat cirrhotic patients. However, appropriate preoperative preparations and meticulous operative techniques are required to reduce blood loss during laparoscopy and even mortality. PMID- 12085148 TI - A prospective comparative study of needlescopic and conventional endoscopic extraperitoneal inguinal hernioplasty. AB - BACKGROUND: Needlescopic inguinal hernioplasty has been made feasible with the miniaturization of instruments and recent advances in laparoscopic surgical technique. Postoperative outcome of needlescopic totally extraperitoneal inguinal hernioplasty (TEP) has not been previously compared with that of conventional TEPs. The objective of the current study is to compare the postoperative outcomes of needlescopic and conventional TEPs. METHODS: From March 1, 2001, to December 30, 2001, a total of 30 patients underwent attempted unilateral needlescopic TEPs. Of these, 12 and 18 patients underwent ambulatory and inpatient procedures, respectively. The results were compared to those of an age-matched cohort of 30 patients who underwent either ambulatory (n = 12) or inpatient (n = 18) conventional TEPs. All data were prospectively collected and analyzed. RESULTS: Needlescopic TEPs were successfully performed in 90% of patients (n = 27). Three procedures were converted to conventional TEPs because of adhesions. Demographic features, hernia types, and mean operative times of the two groups were similar. The mean pain score upon coughing on postoperative day 1 was significantly lower in patients who underwent needlescopic TEPs than in those who had conventional TEPs. Pain scores at rest and upon coughing on days 0 to 6 were otherwise comparable between the two groups. Comparisons of the mean duration of hospitalization, postoperative morbidity, and time taken to resume normal activities showed no significant difference between the two groups. CONCLUSIONS: Needlescopic TEP is a safe technique for the repair of inguinal hernia. Postoperative recovery following needlescopic and conventional TEPs was similar. Needlescopic TEP conferred a significantly lower pain score upon coughing on the first day after operation. PMID- 12085151 TI - The hourglass appearance of the neurofibroma on rectal ultrasound. A case report. AB - Colorectal manifestation of von Recklinghausen's disease is a rare entity. The typical dumbbell shape of the lesion has been described previously, but only from a resected cecal specimen. The authors present the endosonographic appearance of a rectal neurofibroma, which shows a similar hourglass shape, thus facilitating the diagnosis. PMID- 12085149 TI - Laparoscopic physiological hiatoplasty for hiatal hernia: new composite "A" shaped mesh. Physical and geometrical analysis and preliminary clinical results. AB - BACKGROUND: We analyzed, using a theoretical model, the modality of recurrence after a simple cruroplasty for large hiatal hernias, and on the basis of physical and geometrical principles, we conceived a new shaped mesh for a "tension-free" repair. METHODS: We performed a physical and geometrical analysis of the hiatal region on a theoretical model. We also performed an anatomic study on 20 fresh cadavers to verify the reproducibility of the theoretical model and to study the most suitable shape for mesh and its adaptability to the hiatal region. Between September 2000 and October 2001, eight patients received laparoscopic reparation of large (type II or III) hiatal hernias by means of a composite "A"-shaped polytetrafluoroethylene (PTFE)-polypropylene mesh. There were two men and six women; mean age was 65 years (range, 35-78 years). Concomitant esophagitis was found in five patients and impaired esophageal peristalsis in two patients. A total or a partial fundoplication was associated in these cases. RESULTS: The physical and geometrical analysis of the hiatal region explained the reasons for the recurrence after hiatoplasty. The anatomical study on fresh cadavers resulted in a mesh tailored in an "A" shape and permitted to verify the adaptability of such a shaped mesh composed of two layers, polypropylene and PTFE. In the clinical series no conversions occurred; the mortality rate was null. Persistent dysphagia was present in two patients and disappeared after 3 months of treatment. No recurrence was observed at an 8-month average follow-up. CONCLUSION: The preliminary clinical study confirms the feasibility of this tension-free repair and the effectiveness of this composite A-shaped mesh. PMID- 12085152 TI - Intestinal perforation in a parastomal hernia by a migrated plastic biliary stent. AB - Organ perforation is a well-described complication of plastic biliary stent placement [1, 2, 3, 4, 5, 6, 7, 8, 9, 10][1-10]. Most commonly, a stent will cause duodenal perforation while still in the area of the biliary system. This often occurs in the setting of a periampullary diverticulum. Less frequently, a perforation occurs as a result of stent migration into the distal lumenal gastrointestinal tract. These cases have involved a migrated stent impacting at the level of the ileum or a sigmoid colon diverticulum. A plastic biliary stent causing perforation inside an incarcerated small bowel hernia sac has been reported [10]. We describe the first reported case of a plastic biliary stent causing perforation inside a parastomal hernia. PMID- 12085153 TI - Laparoscopic hysterectomy using a computer-enhanced surgical robot. AB - OBJECTIVE: The objective of this study was to describe the technique of laparoscopic hysterectomy using a computer-enhanced robotic surgical system. METHODS: Eleven patients underwent laparoscopic hysterectomy and bilateral salpingo-oophorectomy using a computer-enhanced surgical robot. Four trocars were used: one for the camera, two for the robotic arms controlled by the operating surgeon from the surgeon's console, and an additional port for use by the surgical assistant. RESULTS: Ages ranged from 27 to 77 years, and weight ranged from 54 to 100 kg. Operative time ranged from 4.5 to 10 hours. Estimated blood loss ranged from 50 to 1500 ml. The patients tolerated the procedure and recovered satisfactorily. CONCLUSION: This is the first case series reporting the use of a computer-enhanced surgical robot for performing hysterectomy in humans. It is feasible and well tolerated in this series of patients. As this technology develops, the applications for its use in gynecology and gynecologic oncology will increase. PMID- 12085155 TI - Knot security in laparoscopic surgery. A comparative study with conventional knots. AB - BACKGROUND: Few comparative studies have evaluated conventional and laparoscopic knots. The objective of this study was to evaluate laparoscopic knot reliability and identify which type of knot is most secure. METHODS: Seven types of knots were compared; each one was conventionally tied with four and six throws and similarly by laparoscopy. Dinsmore nomenclature for knots was used. A tension meter was used to evaluate knot reliability, using the loop method, and percentage of failure by slipping and tensile strength was calculated for each group. RESULTS: When S=S=S=S and S=S=S=S=S=S geometry are excluded, there was no difference between laparoscopic and conventional knot. A significant difference between four- and six-throw knots was shown. Excluding S=S=S=S=S=S geometry, all knots in the conventional six-throw group were secure. Intracorporeal 2X1X1X1X1 and 1X1X1X1X1X1 and extracorporeal SXS#SXS#SXS six-throw laparoscopic group knots were secure. CONCLUSIONS: Laparoscopic knots are as secure as conventional knots. All knots must be made with six throws because security is maximized. PMID- 12085157 TI - Abstracts of the British Cancer Research Meeting. 30 June-3 July 2002, Glasgow, United Kingdom. PMID- 12085156 TI - The efficacy and tolerability of risedronate once a week for the treatment of postmenopausal osteoporosis. AB - This study evaluated the efficacy and tolerability of risedronate once a week (35 mg and 50 mg) compared with risedronate 5 mg once daily in women with osteoporosis. We conducted a randomized, double-blind, active-controlled, 2-year study; the primary efficacy assessment was performed after 1 year. Subjects were women aged 50 years or older who had been postmenopausal for at least 5 years, with either a bone mineral density (BMD) T-score of -2.5 or lower (lumbar spine or proximal femur) or a T-score lower than -2 and at least one prevalent vertebral fracture. Subjects received risedronate 5 mg once daily, 35 mg once a week or 50 mg once a week. All subjects also received 1 g daily of elemental calcium supplementation and supplemental vitamin D if the baseline serum levels were low. The primary efficacy measure was percent change in lumbar spine BMD at 12 months. A total of 1,456 women were randomized and received medication; 1,209 (83%) women completed 12 months. The mean percent change (SE) in lumbar spine BMD after 12 months was 4.0% (0.2%) in the 5 mg daily group, 3.9% (0.2%) in the 35 mg group, and 4.2% (0.2%) in the 50 mg group; each once-a-week treatment was determined to be as effective as the daily treatment. Outcomes of the secondary efficacy measurements and safety assessments were also similar in all 3 groups after 12 months. Risedronate 35 mg and 50 mg once a week provide the same efficacy and safety as the daily 5 mg regimen; therefore, the lower dose, 35 mg once a week, is considered optimal for women with postmenopausal osteoporosis who desire a once-a-week regimen. PMID- 12085172 TI - Staging quality is related to the survival of women with endometrial cancer: a Scottish population based study. Deficient surgical staging and omission of adjuvant radiotherapy is associated with poorer survival of women diagnosed with endometrial cancer in Scotland during 1996 and 1997. AB - The association between treatment variation and survival of women with endometrial cancer was investigated. A retrospective cohort based upon the complete Scottish population registered on in-patient and day-case hospital discharge data (Scottish Morbidity Record-1) and cancer registration (Scottish Morbidity Record-6) coded C54 and C55 in ICD10, between 1st January 1996 to 31st December 1997 were analysed. Seven hundred and three patients who underwent surgical treatment out of 781 patients that were diagnosed with endometrial cancer in Scotland during 1996 and 1997. The overall quality of surgical staging was poor. The quality of staging was related to both the year that the surgeon passed the Member of the Royal College of Obstetricians and Gynaecologists examination and also to 'specialist' status but was not related to surgeon caseload. Two clinically important prognostic factors were found to be associated with survival; whether the International Federation of Obstetrics and Gynaecology stage was documented, RHR=2.0 (95% CI=1.3 to 3.1) and also to the use of adjuvant radiotherapy, RHR=2.2 (95% CI=1.5 to 3.5). The associations with survival were strongest in patients with advanced disease, International Federation of Obstetrics and Gynaecology stages 1C through to stage 3. Deficiencies in staging and variations in the use of adjuvant radiotherapy represent a possible source of avoidable mortality in patients with endometrial cancer. Consequently, there should be a greater emphasis on improving the overall quality of surgical staging in endometrial cancer. PMID- 12085173 TI - Adjuvant androgen deprivation impacts late rectal toxicity after conformal radiotherapy of prostate carcinoma. AB - To evaluate whether androgen deprivation impacts late rectal toxicity in patients with localised prostate carcinoma treated with three-dimensional conformal radiotherapy. One hundred and eighty-two consecutive patients treated with 3DCRT between 1995 and 1999 at our Institution and with at least 12 months follow-up were analysed. three-dimensional conformal radiotherapy consisted in 70-76 Gy delivered with a conformal 3-field arrangement to the prostate+/-seminal vesicles. As part of treatment, 117 patients (64%) received neo-adjuvant and concomitant androgen deprivation while 88 (48.4%) patients were continued on androgen deprivation at the end of three-dimensional conformal radiotherapy as well. Late rectal toxicity was graded according to the RTOG morbidity scoring scale. Median follow up is 25.8 (range: 12-70.2 months). The 2-year actuarial likelihood of grade 2-4 rectal toxicity was 21.8+/-3.2%. A multivariate analysis identified the use of adjuvant androgen deprivation (P=0.0196) along with the dose to the posterior wall of the rectum on the central axis (P=0.0055) and the grade of acute rectal toxicity (P=0.0172) as independent predictors of grade 2-4 late rectal toxicity. The 2-year estimates of grade 2-4 late rectal toxicity for patients receiving or not adjuvant hormonal treatment were 30.3+/-5.2% and 14.1+/ 3.8%, respectively. Rectal tolerance is reduced in presence of adjuvant androgen deprivation. PMID- 12085174 TI - Anaplastic thyroid carcinoma: three protocols combining doxorubicin, hyperfractionated radiotherapy and surgery. AB - Patients with anaplastic thyroid carcinoma can rarely be cured, but every effort should be made to prevent death due to suffocation. Between 1984 and 1999, 55 consecutive patients with anaplastic thyroid carcinoma were prospectively treated according to a combined regimen consisting of hyperfractionated radiotherapy, doxorubicin, and when feasible surgery. Radiotherapy was carried out for 5 days a week. The daily fraction until 1988 was 1.0 Gyx2 (A) and 1989-92 1.3 Gyx2 (B). Thereafter 1.6 Gyx2 (C) was administered. Radiotherapy was administered to a total target dose of 46 Gy; of which 30 Gy was administered preoperatively in the first two protocols (A and B), while the whole dose was given preoperatively in the third protocol (C). The therapy was otherwise identical. Twenty mg doxorubicin was administered intravenously weekly. Surgery was possible in 40 patients. No patient failed to complete the protocol due to toxicity. In only 13 cases (24%) was death attributed to local failure. Five patients (9%) 'had a survival' exceeding 2 years. No signs of local recurrence were seen in 33 patients (60%); 5 out of 16 patients in Protocol A, 11 out of 17 patients in Protocol B, 17 out of 22 patients in Protocol C (P=0.017). In the 40 patients undergoing additional surgery, no signs of local recurrence were seen in 5 out of 9 patients, 11 out of 14 patients and 17 out of 17 patients, respectively (P=0.005). PMID- 12085175 TI - Potential role of levocarnitine supplementation for the treatment of chemotherapy induced fatigue in non-anaemic cancer patients. AB - Ifosfamide and cisplatin cause urinary loss of carnitine, which is a fundamental molecule for energy production in mammalian cells. We investigated whether restoration of the carnitine pool might improve chemotherapy-induced fatigue in non-anaemic cancer patients. Consecutive patients with low plasma carnitine levels who experienced fatigue during chemotherapy were considered eligible for study entry. Patients were excluded if they had anaemia or other conditions thought to be causing asthenia. Fatigue was assessed by the Functional Assessment of Cancer Therapy-Fatigue quality of life questionnaire. Treatment consisted of oral levocarnitine 4 g daily, for 7 days. Fifty patients were enrolled; chemotherapy was cisplatin-based in 44 patients and ifosfamide-based in six patients. In the whole group, baseline mean Functional Assessment of Cancer Therapy-Fatigue score was 19.7 (+/-6.4; standard deviation) and the mean plasma carnitine value was 20.9 microM (+/-6.8; standard deviation). After 1 week, fatigue ameliorated in 45 patients and the mean Functional Assessment of Cancer Therapy-Fatigue score was 34.9 (+/-5.4; standard deviation) (P<.001). All patients achieved normal plasma carnitine levels. Patients maintained the improved Functional Assessment of Cancer Therapy-Fatigue score until the next cycle of chemotherapy. In selected patients, levocarnitine supplementation may be effective in alleviating chemotherapy-induced fatigue. This compound deserves further investigations in a randomised, placebo-controlled study. PMID- 12085176 TI - Effective combination chemotherapy with paclitaxel and cisplatin with or without human granulocyte colony-stimulating factor and/or erythropoietin in patients with advanced gastric cancer. AB - A phase II trial was performed to determine the antitumour efficacy and tolerance of combined paclitaxel and cisplatin with or without hematopoetic growth factor support in patients with advanced gastric cancer. Forty-five patients with histologically confirmed metastatic gastric cancer were entered in this trial. Treatment consisted of 2-weekly courses of paclitaxel 160 mg per m2 and cisplatin 60 mg per m2 both given on day 1. Depending on absolute neutrophil counts on the days of scheduled chemotherapeutic drug administration (1000-2000 per microl), a 5-day course of human granulocyte colony-stimulating factor 5 microg x kg(-1) per day was given subcutaneously; in addition, if haemoglobin was <12.0 mg dl(-1), erythropoietin 10 000 IU was administered subcutaneously three times per week. The confirmed overall response rate (intent-to-treat) was 44%, including five complete (11%) and 15 partial remissions (33%). Twelve patients had stable disease (27%), 11 (24%) progressed while on chemotherapy, and two patients were not evaluable. The median time to response was 3 months, the median time to progression 7.0 months, and the median survival time was 11.2 months with 12 patients currently alive. Haematologic toxicity was common, though WHO grade 4 neutropenia occurred in only five patients (11%). Apart from total alopecia in 16 patients (36%), severe non-haematologic adverse reactions included grade 3 peripheral neuropathy in six (13%) and anaphylaxis in two patients. In addition, there was one patient each who experienced grade 3 emesis, diarrhea, and infection, respectively. Our data suggest that the combination of paclitaxel and cisplatin with or without G-CSF and/or erythropoietin has promising therapeutic activity in patients with advanced gastric cancer. PMID- 12085177 TI - Marimastat as maintenance therapy for patients with advanced gastric cancer: a randomised trial. AB - This randomised, double-blind, placebo-controlled study was designed to evaluate the ability of the orally administered matrix metalloproteinase inhibitor, marimastat, to prolong survival in patients with non-resectable gastric and gastro-oesophageal adenocarcinoma. Three hundred and sixty-nine patients with histological proof of adenocarcinoma, who had received no more than a single regimen of 5-fluorouracil-based chemotherapy, were randomised to receive either marimastat (10 mg b.d.) or placebo. Patients were treated for as long as was tolerable. The primary endpoint was overall survival with secondary endpoints of time to disease progression and quality of life. At the point of protocol-defined study completion (85% mortality in the placebo arm) there was a modest difference in survival in the intention-to-treat population in favour of marimastat (P=0.07 log-rank test, hazard ratio=1.23 (95% confidence interval 0.98-1.55)). This survival benefit was maintained over a further 2 years of follow-up (P=0.024, hazard ratio=1.27 (1.03-1.57)). The median survival was 138 days for placebo and 160 days for marimastat, with 2-year survival of 3% and 9% respectively. A significant survival benefit was identified at study completion in the pre defined sub-group of 123 patients who had received prior chemotherapy (P=0.045, hazard ratio=1.53 (1.00-2.34)). This benefit increased with 2 years additional follow-up (P=0.006, hazard ratio=1.68 (1.16-2.44)), with 2-year survival of 5% and 18% respectively. Progression-free survival was also significantly longer for patients receiving marimastat compared to placebo (P=0.009, hazard ratio=1.32 (1.07-1.63)). Marimastat treatment was associated with the development of musculoskeletal pain and inflammation. Events of anaemia, abdominal pain, jaundice and weight loss were more common in the placebo arm. This is one of the first demonstrations of a therapeutic benefit for a matrix metalloproteinase inhibitor in cancer patients. The greatest benefit was observed in patients who had previously received chemotherapy. A further randomised study of marimastat in these patients is warranted. PMID- 12085178 TI - Oxaliplatin plus raltitrexed and leucovorin-modulated 5-fluorouracil i.v. bolus: a salvage regimen for colorectal cancer patients. AB - The aim of the present study was to define the activity and tolerability of a triplet regimen including oxaliplatin 130 mg x m(-2) (2 h i.v. infusion) and raltitrexed 3.0 mg x m(-2) (15 min i.v. infusion) given on day 1, followed by levo-folinic acid 250 mg x m(-2) (2 h i.v. infusion) and 5-fluorouracil 1050 mg x m(-2) i.v. bolus on day 2, every 2 weeks, in pretreated colorectal cancer patients. From April 1999 to December 2000, 50 patients were enrolled: 26 were males and 24 females, their median age was 63 (range, 43-79) years; ECOG performance status was 0 in 26 patients, > or =1 in 24 patients; 26 patients had received previous adjuvant chemotherapy, 40 patients had been exposed to one or two lines of palliative chemotherapy (including irinotecan in 31 cases); 18 patients were considered chemo-refractory. A total of 288 cycles were administered, with a median number of 6 (range 1-12) courses per patient. A complete response was obtained in three patients, and a partial response in nine patients, giving a major response rate of 24% (95% confidence interval, 13-38%), while 15 further patients showed a stable disease, for an overall control of tumour growth in 60% of patients. Three complete responses and three partial responses were obtained in patients pretreated with irinotecan (response rate, 19%); among refractory patients, three achieved partial responses (response rate, 13%). After a median follow-up of 18 (range, 10-30) months, 40 patients showed a progression of disease: the growth modulation index ranged between 0.2 and 2.5: it was > or =1.33 (showing a significant delay of tumour growth) in 16 (40%) patients. Actuarial median progression-free survival time was 7.6 months, and median survival time was 13.6 months: estimated probability of survival was 55% at 1 year. Main severe toxicity was neutropenia: World Health Organisation grade 4 affected 32% of patients; non-haematological toxicity was mild: World Health Organisation grade 3 diarrhoea was complained of by 8%, and grade 3 stomatitis by 4% of patients; neurotoxicity (according to Levi scale) was scored as grade 3 in 8% of patients. In conclusion, this regimen was manageable and active as salvage treatment of advanced colorectal cancer patients; it showed incomplete cross resistance with irinotecan-based treatments, and proved to delay the progression of disease in a relevant proportion of treated patients. PMID- 12085179 TI - Tissue levels of active matrix metalloproteinase-2 and -9 in colorectal cancer. AB - The bioactivity of matrix metalloproteinases was studied in tissues from colorectal cancer patients by means of both quantitative gelatin zymography and a fluorometric activity assay. Next to paired samples of tumour tissue and distant normal mucosa (n=73), transitional tissue was analysed from a limited (n=33) number of patients. Broad-spectrum matrix metalloproteinase activity and both the active and latent forms of the gelatinases matrix metalloproteinase-2 and -9 were higher in tumour than in normal mucosa. The ratio's between active and latent forms of matrix metalloproteinase-2 and -9 were highest in tumour tissue and normal mucosa, respectively. Matrix metalloproteinase-2 levels, both active and latent forms, correlated inversely with stage of disease, the tumours without synchronous distant metastases containing significantly (P=0.005) more active matrix metalloproteinase-2 than the others. At much lower levels of activity, the same trend was observed in distant normal mucosa. The level of latent form of matrix metalloproteinase-9 in tumour depended on tumour location. Neither the active form of matrix metalloproteinase-9 nor broad-spectrum matrix metalloproteinase activity in tumour tissue did correlate with any of the clinicopathological parameters investigated. The results demonstrate explicit differences between the activity of matrix metalloproteinase-2 and -9, indicating different roles for both gelatinases in tumour progression. Such data are necessary in order to develop rational anti-cancer therapies based on inhibition of specific matrix metalloproteinases. PMID- 12085180 TI - Cytokeratin 7 and 20 staining for the diagnosis of lung and colorectal adenocarcinoma. AB - The origin of metastatic adenocarcinoma lesions can sometimes be difficult to diagnose. The objectives of our study were to establish the cytokeratin staining pattern of primary and metastatic lung and colorectal adenocarcinomas, and to determine if this helps to identify the site of origin of metastatic lesions. We reviewed a total of 102 tissue samples from patients in our tumour registry, with either primary or metastatic lung or colorectal adenocarcinoma. Tissue sections were stained for cytokeratin 7 and 20 and read as positive or negative for staining. Clinical and radiologic information was reviewed from computerised charts. The cytokeratin 7+/cytokeratin 20- pattern characterised 96% (29 out of 30) of primary and 95% (21 out of 22) of metastatic lung adenocarcinomas. All the primary (26), and 88% (21 out of 24) of metastatic colorectal adenocarcinomas stained cytokeratin 7-/cytokeratin 20+. Samples from a variety of metastatic sites were evaluated for cytokeratin 7 and 20 staining. Out of the 102 samples, in 95% (97 out of 102) of the cases, the cytokeratin 7 and cytokeratin 20 staining pattern characterised and differentiated between lung and colorectal adenocarcinoma. Primary and metastatic lung adenocarcinomas show a cytokeratin 7+/cytokeratin 20- staining pattern, while colorectal adenocarcinomas stain cytokeratin 7-/cytokeratin 20+. Cytokeratin staining is helpful in the diagnostic differentiation of metastatic lesions from these two common primaries, and assists in determining the site of origin of metastatic lesions. PMID- 12085181 TI - Inactivation of O6-methylguanine-DNA methyltransferase by promoter CpG island hypermethylation in gastric cancers. AB - Promoter hypermethylation of CpG islands in tumour suppressor genes can lead to transcriptional inactivation. To investigate the association between methylation and expression at O6-methylguanine-DNA methyltransferase, we performed methylation-specific PCR and immunohistochemistry in 149 gastric carcinomas. Promoter methylation was found in 14.1% of tumours and loss of expression was detected in 11.4% of tumours. To examine correlation between the O6-methylguanine DNA methyltransferase expression and the clinical data, we investigated O6 methylguanine-DNA methyltransferase expression in 315 consecutive gastric carcinomas. A similar frequency of loss of O6-methylguanine-DNA methyltransferase expression was confirmed in these cases. The loss of O6-methylguanine-DNA methyltransferase expression was significantly associated with pTNM stage (P=0.037), tumour invasion (P=0.02), microsatellite instability (P=0.041) and overall survival (P=0.01). Among 11 gastric cancer cell lines, SNU-620 showed the loss of O6-methylguanine-DNA methyltransferase expression as well as promoter methylation. After treatment with 5-aza-2-deoxycytidine, a demethylating agent, SNU-620 re-expressed O6-methylguanine-DNA methyltransferase mRNA. In summary, we suggest that during gastric carcinogenesis, the loss of O6-methylguanine-DNA methyltransferase expression frequently occurs via the hypermethylation of the CpG islands of the promoter region, and that this is significantly associated with the clinicopathological characteristics. PMID- 12085182 TI - Phosphorylated tyrosine-containing proteins in primary lung cancer correlates with proliferation and prognosis. AB - To determine the usefulness of tyrosine phosphorylation in evaluating biological characteristics, we attempted to evaluate the relationship between the amount of phosphorylated tyrosine-containing proteins and clinicopathological factors, cell proliferation and outcome in non-small cell lung cancer. To evaluate phosphorylated tyrosine-containing proteins we used 96 surgically resected materials of non-small cell lung cancer and normal peripheral lung, while immunohistochemical evaluation was performed. Cell proliferating ability was evaluated using the labelling index of proliferating cell nuclear antigen positive nuclear staining cells. There were statistically significant differences between the expression levels of phosphorylated tyrosine-containing proteins of normal and cancerous tissues (P<0.0001). Evaluations based on clinicopathological factors apart from histopathological differentiation, showed no statistically significant differences of phosphorylated tyrosine-containing proteins expression. However, phosphorylated tyrosine-containing proteins correlated with cell proliferation activity evaluated (P((Low, High))<0.0001; P((Low, Int)) <0.0001; P((Int, High))<0.0001). Furthermore, non-small cell lung cancer cases with high expression and intermediate expression of phosphorylated tyrosine containing proteins had a significantly shorter disease-free postoperative survival than those with low expression of phosphorylated tyrosine-containing proteins using log-rank analysis (P((Low, Int)) <0.0028; P((Low, High))=0.0002). Furthermore, phosphorylated tyrosine-containing proteins expression level statistically contributed to disease-free survival in Cox's proportional hazard model. Therefore, phosphorylated tyrosine-containing proteins in non-small cell lung cancer tissues seem to reflect its biological malignancy, and this evaluation may be valuable for constructing the most appropriate therapeutic strategy. PMID- 12085183 TI - The prognostic influence of bcl-2 in malignant glioma. AB - The bcl-2 gene is one of a complex group of genes which control programmed cell death. Bcl-2 acts to extend cell survival by blocking apoptosis, and thereby may influence tumour prognosis. This study of 187 high grade gliomas reviews clinicopathological prognostic features and the relationship to bcl-2 expression. Bcl-2 immunostaining was assessed in 159 specimens from these patients, by scoring systems of 0 to 3 for intensity of scoring and proportion of cells staining. Age, histology, pre- and post-operative performance status were found to be strongly predictive of survival (log rank test P<0.0001). The type of surgery performed did not influence survival in this group of patients. The expression of bcl-2 had a significant relationship with survival (univariate Cox model P=0.0302, hazard ratio 0.8, 95% confidence interval 0.65-0.98), with increased staining associated with improved survival. Multivariate analysis showed performance status, histology and proportion of cells staining for bcl-2 to be independently predictive of survival. Bcl-2 staining was not related to histological grade of tumours. PMID- 12085184 TI - Tumour microvessel density as predictor of chemotherapy response in breast cancer patients. AB - The aim of this study was to evaluate the predictive value of intratumoural microvessel density in breast cancer. We studied immunohistochemically primary tumours of 104 patients with metastasised breast cancer who took part in a randomised multicentre trial comparing docetaxel to sequential methotrexate and 5 fluorouracil. Vessels were highlighted with factor VIII staining and counted microscopically. Microvessel density was compared with clinical response to chemotherapy and patient survival. The microvessel density of the primary tumour was not significantly associated with patient's response to chemotherapy, time to progression or overall survival in the whole patient population or in the docetaxel or methotrexate and 5-fluorouracil groups. However, disease-free survival was longer in patients with low microvessel density (P=0.01). These findings suggest that microvessel density of the primary tumour cannot be used as a predictive marker for chemotherapy response in advanced breast cancer. PMID- 12085185 TI - Expression of cdc25A and cdc25B proteins in thyroid neoplasms. AB - Cdc25B and cdc25A phosphates are prominent stimulators of cell cycle progression and recent studies have also suggested their oncogenic roles. To elucidate the role of these proteins in thyroid neoplasms, we immunohistochemically investigated their expression, and neither protein was expressed in normal follicular cells. Cdc25B was frequently overexpressed in follicular adenoma and minimally invasive follicular carcinoma, but the incidence was significantly lower in widely invasive follicular carcinoma. Furthermore, the cdc25B expression level significantly decreased with the dedifferentiation of thyroid carcinoma. Cdc25A overexpression was observed in high incidences in all types of thyroid neoplasms. These results suggest that cdc25B and cdc25A play oncogenic roles in thyroid follicules and that cdc25B works predominantly in the early phase of the progression of thyroid carcinoma, whereas cdc25A plays a fundamental role in the development of thyroid neoplasms. PMID- 12085186 TI - Hypoxia enhances the expression of autocrine motility factor and the motility of human pancreatic cancer cells. AB - The incidence of distant metastases is higher in the tumours with low oxygen pressure than in those with high oxygen pressure. It is well known that hypoxia induces the transcription of various genes involved in angiogenesis and anaerobic metabolism necessary for the growth of tumour cells in vivo, suggesting that hypoxia may also induce the transcription of metastasis-associated genes. We sought to identify the metastasis-associated genes differentially expressed in tumour cells under hypoxic conditions with the use of a DNA microarray system. We found that hypoxia enhanced the expression of autocrine motility factor mRNA in various cancer cells and also enhanced the random motility of pancreatic cancer cells. Autocrine motility factor inhibitors abrogated the increase of motility under hypoxic conditions. In order to explore the roles of hypoxia-inducible factor-1alpha, we established hypoxia-inducible factor-1alpha-transfectants and dominant negative hypoxia-inducible factor-1alpha-transfectants. Transfection with hypoxia-inducible factor-1alpha and dominant-negative hypoxia-inducible factor-1alpha enhanced and suppressed the expression of autocrine motility factor/phosphohexase isomerase/neuroleukin mRNA and the random motility, respectively. These results suggest that hypoxia may promote the metastatic potential of cancer cells through the enhanced autocrine motility factor/phosphohexase isomerase/neuroleukin mRNA expression and that the disruption of the hypoxia-inducible factor-1 pathway may be an effective treatment for metastasis. PMID- 12085187 TI - Somatic glypican 3 (GPC3) mutations in Wilms' tumour. AB - Tumour and normal tissue from 41 male cases of Wilms' tumour were screened to determine the presence of sequence variants in the glypican 3 (GPC3) gene. Two non-conservative single base changes were present in tumour tissue only. These findings imply a possible role for GPC3 in Wilms' tumour development. PMID- 12085188 TI - Overexpression of Betaig-h3 gene downregulates integrin alpha5beta1 and suppresses tumorigenicity in radiation-induced tumorigenic human bronchial epithelial cells. AB - Interaction between cell and extracellular matrix plays a crucial role in tumour invasion and metastasis. Using an immortalised human bronchial epithelial (BEP2D) cell model, the study here shows that expression of Betaig-h3 gene, which encodes a secreted adhesion molecule induced by transforming growth factor-beta, is markedly decreased in several independently generated, radiation-induced tumour cell lines (TL1-TL5) relative to parental BEP2D cells. Transfection of Betaig-h3 gene into tumour cells resulted in a significant reduction in tumour growth. While integrin receptor alpha5beta1 was overexpressed in tumour cells, its expression was corrected to the level found in control BEP2D cells after Betaig h3 transfection. These data suggest that Betaig-h3 gene is involved in tumour progression by regulating integrin receptor alpha5beta1. The findings provide strong evidence that the Betaig-h3 gene has tumour suppressor function in human BEP2D cell model and suggest a potential target for interventional therapy. PMID- 12085189 TI - Association of RET codon 691 polymorphism in radiation-induced human thyroid tumours with C-cell hyperplasia in peritumoural tissue. AB - The RET proto-oncogene encodes a protein structurally related to transmembrane receptors with an intracellular tyrosine kinase domain. In human thyroid gland, the RET proto-oncogene is normally expressed in parafollicular C-cells. Thyroid C cell hyperplasia is associated with inherited medullary thyroid carcinomas and is considered as a pre-neoplastic stage of C-cells disease. It has also been observed in thyroid tissues adjacent to follicular and papillary carcinomas. In order to study the relationship between a misfunctioning of the RET proto oncogene and the presence of C-cell hyperplasia, we compared a series of thyroid glands presenting sporadic or radiation-associated tumours, as well as samples of unrelated normal thyroid tissues, for alteration in exons 10 and 11 of the gene and for the presence or absence of C-cell hyperplasia. Here we report a significantly higher frequency of C-cell hyperplasia present in peritumoural thyroid tissues of radiation-induced epithelial thyroid tumours, than in peritumoural of sporadic thyroid tumours or in control normal thyroid tissues (P=0.001). A G691S RET polymorphism was present with a higher frequency in radiation-induced epithelial thyroid tumours (55%) than in sporadic tumours (20%) and in control normal thyroid tissues (15%). Interestingly, this polymorphism was associated in the majority (88%) of radiation-induced tumours with a C-cell hyperplasia in the peritumoural tissues. Several explanations for this association are discussed. PMID- 12085190 TI - Induction of endothelial cell apoptosis by the antivascular agent 5,6 Dimethylxanthenone-4-acetic acid. AB - 5,6-Dimethylxanthenone-4-acetic acid, synthesised in this laboratory, reduces tumour blood flow, both in mice and in patients on Phase I trial. We used TUNEL (TdT-mediated dUTP nick end labelling) assays to investigate whether apoptosis induction was involved in its antivascular effect. 5,6-Dimethylxanthenone-4 acetic acid induced dose-dependent apoptosis in vitro in HECPP murine endothelial cells in the absence of up-regulation of mRNA for tumour necrosis factor. Selective apoptosis of endothelial cells was detected in vivo in sections of Colon 38 tumours in mice within 30 min of administration of 5,6 Dimethylxanthenone-4-acetic acid (25 mg x kg(-1)). TUNEL staining intensified with time and after 3 h, necrosis of adjacent tumour tissue was observed. Apoptosis of central vessels in splenic white pulp was also detected in tumour bearing mice but not in mice without tumours. Apoptosis was not observed in liver tissue. No apoptosis was observed with the inactive analogue 8-methylxanthenone-4 acetic acid. Positive TUNEL staining of tumour vascular endothelium was evident in one patient in a Phase I clinical trial, from a breast tumour biopsy taken 3 and 24 h after infusion of 5,6-Dimethylxanthenone-4-acetic acid (3.1 mg x m(-2)). Tumour necrosis and the production of tumour tumour necrosis factor were not observed. No apoptotic staining was seen in tumour biopsies taken from two other patients (doses of 3.7 and 4.9 mg x m(-2)). We conclude that 5,6 Dimethylxanthenone-4-acetic acid can induce vascular endothelial cell apoptosis in some murine and human tumours. The action is rapid and appears to be independent of tumour necrosis factor induction. PMID- 12085191 TI - Increased expression of beta 2-microglobulin in multidrug-resistant tumour cells. AB - The rat monoclonal antibody LMR-12 was shown earlier to react with a plasma membrane protein, upregulated in multidrug-resistant cell lines. In this study, we observed distinct LMR-12 staining in 36 out of 55 non-drug-selected tumour cell lines, including melanomas, renal cell-, colon- and lung carcinomas, whereas in other tumour types, such as leukaemia and ovarian cancer, LMR-12 staining was generally low or absent. The cDNA encoding the LMR-12 antigen was isolated from a library of the multidrug-resistant human fibrosarcoma cell line HT1080/DR4 by expression cloning in MOP8 cells. Sequence analysis showed that the LMR-12 antigen is identical to the major histocompatibility complex class I molecule beta 2-microglobulin (beta2-m). The LMR-12/ beta2-m staining results were confirmed by mRNA microarray data from an independent National Cancer Institute study, as well as by newly obtained reverse transcriptase polymerase chain reaction data. Further analysis of the microarray data showed that beta2-m levels closely reflected levels of major histocompatibility complex class I heavy chains and the transporter associated with antigen processing. Since the ABC transporter associated with antigen processing was previously shown to contribute to multidrug-resistance, it may very well be that the observed LMR-12/ beta2-m levels are secondary to (elevated) levels of the transporter associated with antigen processing. A perspective arising from the present study is that drug resistant tumour cells may, by having elevated levels of major histocompatibility complex related molecules, be particular good candidates for alternative therapeutic therapies, such as cytotoxic T cell mediated immune-therapies. PMID- 12085192 TI - Implication of mitochondria-derived ROS and cardiolipin peroxidation in N-(4 hydroxyphenyl)retinamide-induced apoptosis. AB - We have studied the effect of N-(4-hydroxyphenyl)retinamide on either malignant human leukaemia cells or normal cells and investigated its mechanism of action. We demonstrate that 4HPR induces reactive oxygen species increase on mitochondria at a target between mitochondrial respiratory chain complex I and II. Such oxidative stress causes cardiolipin peroxidation which in turn allows cytochrome c release to cytosol, caspase-3 activation and therefore apoptotic consumption. Moreover, this apoptotic pathway seems to be bcl-2/bax independent and count only on malignant cells but not normal nor activated lymphocytes. PMID- 12085194 TI - Differential interactions between IGFBP-3 and transforming growth factor-beta (TGF-beta) in normal vs cancerous breast epithelial cells. AB - In addition to modulating insulin-like growth factors action, it is now clear that insulin-like growth factor-binding protein-3 also has intrinsic effects on cell growth and survival. We have compared the effects of insulin-like growth factor-binding protein-3 and transforming growth factor-beta on cell proliferation and death of Hs578T cells and the normal breast epithelial cell line, MCF-10A. The growth of MCF-10A cells was inhibited at low concentrations of insulin-like growth factor-binding protein-3 but stimulated at high concentrations. These differential effects were unaffected in the presence of an insulin-like growth factor-I receptor antagonist. A synthetic peptide corresponding to the serine phosphorylation domain of insulin-like growth factor binding protein-3 (that does not bind to insulin-like growth factors) also mimicked these differential actions. The growth of both cell lines was significantly inhibited by transforming growth factor-beta, this was associated with a 14-fold increase of insulin-like growth factor-binding protein-3 secreted by the Hs578T cells but a five-fold decrease of insulin-like growth factor binding protein-3 secreted by MCF-10A cells. Replacement doses of exogenous insulin-like growth factor-binding protein-3 overcame the transforming growth factor-beta-induced growth inhibition in the MCF-10A cells. Cell death induced by ceramide was significantly reduced by insulin-like growth factor-binding protein 3 in the MCF-10A cells and depleting insulin-like growth factor-binding protein-3 with transforming growth factor-beta in these cells consequently increased their susceptibility to ceramide. In contrast, insulin-like growth factor-binding protein-3 enhanced apoptosis induced by ceramide in the Hs578T cells but transforming growth factor-beta treated Hs578T cells were resistant to apoptosis. The addition of anti-sense mRNA to insulin-like growth factor-binding protein-3 significantly abrogated this effect of transforming growth factor-beta. These data indicate that insulin-like growth factor-binding protein-3 has intrinsic activity capable of inhibiting or enhancing the growth and survival of breast epithelial cells depending on the cell line and exposure to other cytokines. PMID- 12085193 TI - Decreased proliferation of human melanoma cell lines caused by antisense RNA against translation factor eIF-4A1. AB - Control of translation initiation was recognised as a critical checkpoint for cell proliferation and tumorigenesis. In human melanoma cells, we have previously reported consistent overexpression of translation initiation factor eIF-4A1. Here, we investigated by transfection of antisense constructs its significance for the control of melanoma cell growth. The tetracycline-inducible expression system was established in melanoma cells, and three fragments of the 5'-, central , and 3'-portion of the eIF-4A1 cDNA were subcloned in antisense and in sense orientation after a tetracycline inducible promoter. Significant proliferation decrease was obtained after transient transfection and induction of antisense RNA directed against the 5'- and the central portion (up to 10%), whereas, no effects were seen after induction of the 3'-fragment and the sense controls. Cell clones stably transfected with the central antisense fragment revealed after doxycycline induction reduced expression of endogeneous eIF-4A1 mRNA correlated with decreased proliferation rates (up to 6%). These data demonstrate the applicability of antisense strategies against translation factors in melanoma cells. Translation initiation factor eIF-4A1 contributes to the control of melanoma cell proliferation and may be taken into consideration when scheduling new therapeutic approaches targeting the translational control. PMID- 12085195 TI - Is the ratio of pleural mesothelioma mortality to pleural cancer mortality approximately unity for Italy? Considerations from the oldest regional mesothelioma register in Italy. PMID- 12085199 TI - Generation of dendritic cell-based vaccines for cancer therapy. AB - Dendritic cells play a major role in the generation of immunity against tumour cells. They can be grown under various culture conditions, which influence the phenotypical and functional properties of dendritic cells and thereby the consecutive immune response mainly executed by T cells. Here we discuss various conditions, which are important during generation and administration of dendritic cells to elicit a tumouricidal T cell-based immune response. PMID- 12085200 TI - Autologous cell vaccine as a post operative adjuvant treatment for high-risk melanoma patients (AJCC stages III and IV). The new American Joint Committee on Cancer. AB - This study evaluates the overall survival and disease free survival of melanoma patients that were treated with an autologous melanoma cell vaccine, administered as a post-operative adjuvant. Included are 43 patients with totally resected metastatic melanoma (28-AJCC stage III, 15-AJCC stage IV), with a median follow up of 34 months (6-62). The treatment consisted of eight doses of a vaccine made of 10-25x10(6) autologous melanoma cells either released from the surgical specimen or grown in cell cultures. Tumour cells were conjugated with hapten dinitrophenyl, mixed with Bacille Calmette Guerin and irradiated to 110 Gy. Both disease free survival and overall survival were found to be correlated with intensity of evolving delayed type hypersensitivity to subcutaneous injection of unmodified melanoma cells. Patients with a delayed type hypersensitivity reaction of > or =10 mm had a median disease free survival of 17 months (mean 35 months) and a mean overall survival of 63 months (median not reached). In contrast, patients with a negative or weak delayed type hypersensitivity had a median disease free survival of 9 months (relative risk of recurrence=4.5, P=0.001), and a median overall survival of 16 months (relative risk of death=15, P=0.001). Stage III patients with a positive delayed type hypersensitivity reaction had an improved disease free survival of 16 months and a mean overall survival of 38 months, whereas patients with a negative delayed type hypersensitivity had a median disease free survival of 7 months (relative risk=4.5, P=0.02) and a median overall survival of 16 months (relative risk=9.5, P=0.005). The adjuvant administration of autologous melanoma vaccine was associated with improved disease-free and overall survival to selected patients who successfully attained anti-melanoma reactivity as detected by positive delayed type hypersensitivity reactions to unmodified melanoma cells. PMID- 12085201 TI - The unrecognised cost of cancer patients' unrelieved symptoms:a nationwide follow up of their surviving partners. AB - We investigated if a cancer patient's unrelieved symptoms during the last 3 months of life increase the risk of long-term psychological morbidity of the surviving partner. All women (n=506) living in Sweden under 80 years of age, who lost their husband/partner owing to cancer of the prostate in 1996 or of the urinary bladder in 1995 or 1996 were asked to answer an anonymous postal questionnaire, 2-4 years after their loss. The widows' psychological morbidity was associated with the patient's unrelieved mental symptoms. When the patient was perceived to have been very anxious during last three months of life (compared to no observed symptoms) the relative risks for the widows' psychological morbidity were: 2.5 (1.4-4.3) for depression and 3.4 (1.4-8.2) for anxiety. When comparing reports of the patient's pain (much vs no), the relative risks were 0.8 (0.5-1.2) for widowhood depression, and 0.8 (0.4-1.7) for widowhood anxiety. The patients were found to have had adequate access to physical pain control but poor access to psychological symptom control. Efficiency in diagnosing and treating psychological complications of terminally ill cancer patients may not only improve their quality of life but possibly also prevent long-term psychological morbidity of their surviving partners. PMID- 12085202 TI - Tamoxifen effects on subjective and psychosexual well-being, in a randomised breast cancer study comparing high-dose and standard-dose chemotherapy. AB - To evaluate the impact of tamoxifen on subjective and psychosexual well-being in breast cancer patients in relation to type of prior chemotherapy and menopausal status. Longitudinal interview study in breast cancer patients during and after adjuvant tamoxifen use. Menopausal status was defined by last menstrual period and serum oestradiol and FSH levels. Gynaecology outpatient clinic, Tertiary Referral Hospital, January 1995 to September 1999. Breast cancer patients <56 years of age, participating in a randomised trial comparing adjuvant high-dose (n=45) and standard-dose (n=53) chemotherapy, followed by radiotherapy and tamoxifen. Relative incidence and correlation of subjective and psychosexual symptoms during and after tamoxifen. During tamoxifen the most frequent complaints were hot flushes (85%), disturbed sleep (55%), vaginal dryness and/or dyspareunia (47%), decreased sexual desire (44%) and musculo-skeletal symptoms (43%). Disturbed sleep correlated with hot flushes (P<0.0005) and concentration problems (P<0.05). Decreased sexual interest correlated with vaginal dryness (P<0.0005) and/or dyspareunia (P<0.0005). In the high-dose group more patients became postmenopausal (95% vs 33%) and more patients reported symptoms than in the standard-dose group (P<0.05). After discontinuation of tamoxifen, symptoms decreased significantly. However, hot flushes, disturbed sleep and vaginal dryness persisted more often in patients who remained postmenopausal after high dose chemotherapy (P<0.05). Overall, during tamoxifen patients reported many symptoms. More patients become postmenopausal after high-dose chemotherapy, and they remain often symptomatic after tamoxifen. PMID- 12085203 TI - A phase I trial of weekly gemcitabine and concurrent radiotherapy in patients with locally advanced pancreatic cancer. AB - This study investigated the maximum-tolerated dose of gemcitabine based on the frequency of dose-limiting toxicities of weekly gemcitabine treatment with concurrent radiotherapy in patients with locally advanced pancreatic cancer. Fifteen patients with locally advanced pancreatic cancer that was histologically confirmed as adenocarcinoma were enrolled in this phase I trial of weekly gemcitabine (150-350 mg x m(-2)) with concurrent radiotherapy (50.4 Gy in 28 fractions). Gemcitabine was administered weekly as an intravenous 30-min infusion before radiotherapy for 6 weeks. Three of six patients at the dose of 350 mg x m( 2) of gemicitabine demonstrated dose-limiting toxicities involving neutropenia/ leukocytopenia and elevated transaminase, while nine patients at doses of 150 mg x m(-2) and 250 mg x m(-2) did not demonstrate any sign of dose-limiting toxicity. Of all 15 enrolled patients, six patients (40.0%) showed a partial response. More than 50% reduction of serum carbohydrate antigen 19-9 level was observed in 13 (92.9%) of 14 patients who had pretreatment carbohydrate antigen 19-9 levels of 100 U x ml(-1) or greater. The maximum-tolerated dose of weekly gemcitabine with concurrent radiotherapy was 250 mg x m(-2), and this regimen may have substantial antitumour activity for patients with locally advanced pancreatic cancer. A phase II trial of weekly gemcitabine at the dose of 250 mg x m(-2) with concurrent radiation in patients with locally advanced pancreatic cancer is now underway. PMID- 12085204 TI - Alternating dose-dense chemotherapy in patients with high volume disseminated non seminomatous germ cell tumours. AB - Only about half of patients with a poor-prognosis non-seminomatous germ-cell tumours can achieve a cure. The aim of this phase II study was to assess the efficacy and toxicity of a dose-dense alternating chemotherapy regimen in this subset of patients. High volume non-seminomatous germ-cell tumours was defined as follows: at least two sites of non pulmonary metastases, an extragonadal primary tumour, a serum human chorionic gonadotropin level higher than 10 000 mIU x ml( 1), or a alpha-foetoprotein level higher than 2000 mIU ml(-1). Patients who fulfilled these criteria were treated with the so-called BOP-CISCA-POMB-ACE regimen (bleomycin, vincristine, and cisplatin; cisplatin, cyclophosphamide, and doxorubicin; cisplatin, vincristine, methotrexate, and bleomycin; etoposide, dactinomycin, and cyclophosphamide) plus granulocyte colony-stimulating factor. A total of 58 patients were enrolled. Patients were retrospectively classified according to the International Germ-Cell Cancer Consensus Group classification; 38 patients (66%) had poor-prognosis disease and 19 patients (33%) had intermediate-prognosis. Patients received a median of 2.5 courses (range 0.25 to five courses) of the BOP-CISCA-POMB-ACE regimen. Forty-two patients (72.4%) had a complete response to therapy. With a median follow-up time of 31 months, the 3 year progression-free survival rate was 71% (95% confidence interval, 60 to 84%) and the 3-year overall survival rate was 73% (95% confidence interval: 62 to 86%). The 3-year PFS rates were 83% (95% confidence interval: 68 to 100%) in the intermediate-prognosis group and 65% (95% confidence interval: 51 to 82%) in the poor-prognosis group. Early side effects included mainly grade 4 haematologic toxicity (neutropaenia in 79% of patients, thrombocytopaenia in 69%, anaemia in 22%), grade 4 stomatitis (19%), and four early deaths (7% of patients), at least partially related to toxicity. The dose-dense BOP-CISCA-POMB-ACE regimen is highly active in patients with non-seminomatous germ-cell tumours classified as intermediate-prognosis or poor-prognosis according to the International Germ-Cell Cancer Consensus Group. Because outcomes with this regimen compare favourably with outcome after standard therapy, dose-dense chemotherapy should be further investigated in this subset of patients. PMID- 12085205 TI - Apoptosis control and proliferation marker in human normal and neoplastic adrenocortical tissues. AB - We evaluated by immunohistochemistry the expression of the Bcl-2 and p53 proteins, as markers of apoptosis control, and of MIB-1, as a marker of cell proliferation, in a series of normal and neoplastic adrenocortical tissues. The specimens were 13 normal adrenals, 13 aldosterone-producing adenomas, 13 non functioning adenomas and 16 carcinomas. Results were calculated as percentage of immunostained cells by using specific antibodies. No p53 protein was detected in any of the adrenocortical adenomas (functioning and non functioning) or normal adrenals, while p53 was overexpressed in 15 out of 16 carcinomas. In particular, 10 adrenal cancer specimens (62.5%) showed strong staining in a high percentage (range 10-50%) of the malignant cells. The percentage of Bcl-2 positive cells was higher (P<0.05 or less) in non-functioning adenomas (8.1+/-1.9%) and in carcinomas (14.9+/-5.6%) than in normals (2.9+/-0.9%) and aldosterone-producing adenomas (5.3+/-1.3%) since four specimens of the non-functioning adenomas-group (30.7%) and six of the carcinomas-group (37.5%) showed over 10% positivity (cut off for normal values, set at 90th percentile of our controls). MIB-1 positivity was 0.50+/-0.36% in normals, 0.54+/-0.08% in non-functioning adenomas and 0.54+/ 0.08% in aldosterone-producing adenomas. MIB-1 was expressed in all carcinomas with values (13.7+/-3.1%) significantly (P<0.0006) higher than in the other groups. In conclusion, the present data indicate that the apoptosis control and proliferation activity evaluated by the p53 and MIB-1 proteins are impaired in adrenal carcinomas but preserved in adenomas, independently of their functional status. Therefore, these immunohistochemical markers, overexpressed in carcinomas only, may be useful in the diagnosis of malignancy in adrenocortical tumours. Whether Bcl-2 positivity found in some carcinomas and non-functioning adenomas may constitute, in the latter, a negative prognostic marker is still unknown. PMID- 12085207 TI - Expression of multidrug resistance-associated protein1,P-glycoprotein, and thymidylate synthase in gastric cancer patients treated with 5-fluorouracil and doxorubicin-based adjuvant chemotherapy after curative resection. AB - Both 5-fluorouracil and doxorubicin are commonly used agents in chemotherapy of gastric cancer in adjuvant setting as well as metastatic disease. In a variety of malignancies, high expression of multidrug resistance-associated protein1 and P glycoprotein has been associated with resistance to doxorubicin, whereas 5 fluorouracil resistance has correlated with the level of thymidylate synthase expression. We evaluated the expression of multidrug resistance-associated protein1, P-glycoprotein, and thymidylate synthase using immunohistochemistry in 103 locally advanced gastric cancer patients (stage IB-IV) who underwent 5 fluorouracil and doxorubicin-based adjuvant chemotherapy after curative resection and investigated the association between their expression and clinicopathologic characteristics including prognosis of the patients. While high expression (> or =5% of tumour cells positive) of multidrug resistance-associated protein1 and P glycoprotein was observed in 70 patients (68%) and 42 patients (41%), respectively, 65 patients (63%) had primary tumours with high expression (> or =25% of tumour cells positive) of thymidylate synthase. There was a significant association between multidrug resistance-associated protein1 and P-glycoprotein expression (P<0.0001) as well as P-glycoprotein and thymidylate synthase expression (P<0.0001). High multidrug resistance-associated protein1 and P glycoprotein expressions were associated with well and moderately differentiated histology (P<0.0001 and P=0.03, respectively) and intestinal type (P<0.0001 and P=0.009, respectively). High multidrug resistance-associated protein1 expression correlated with lymph node metastasis (P=0.037), advanced stage (P=0.015), and older age (P=0.021). Five-year disease-free survival and overall survival of total patients were 55.2% and 56.2%, respectively, with a median follow-up of 68 months. There were no significant differences in disease-free survival and overall survival according to the expression of multidrug resistance-associated protein1 (P=0.902 and P=0.975, respectively), P-glycoprotein (P=0.987 and P=0.955, respectively), and thymidylate synthase (P=0.604 and P=0.802, respectively). Concurrent high expression of these proteins (high multidrug resistance-associated protein1/P-glycoprotein, high multidrug resistance associated protein1/thymidylate synthase, high P-glycoprotein/thymidylate synthase) did not correlate with disease-free survival or overall survival. Even high expression of all three proteins was not associated with poor disease-free survival (P=0.919) and overall survival (P=0.852). In conclusion, high expression of multidrug resistance-associated protein1, P-glycoprotein, and thymidylate synthase did not predict poor prognosis of gastric cancer patients treated with 5 fluorouracil and doxorubicin-based adjuvant chemotherapy. A larger study including patients treated with surgical resection alone would be necessary. PMID- 12085209 TI - Investigations on a clinically and functionally unusual and novel germline p53 mutation. AB - This report describes an individual with a rare choroid plexus papilloma in adulthood (age 29) after earlier having an osteosarcoma (age 22). The results from this study, and others, suggest that it may be advisable to consider the possibility of a germline p53 mutation in adults presenting with choroid plexus tumours. In the current study automated DNA sequencing of genomic DNA detected a novel germline 7 base pair insertion in exon 5 of the p53 gene in this patient. The alteration in frame would produce amino acid substitutions beginning with alanine to glycine at position 161 and a stop codon at position 182 in the mutated protein. Surprisingly two assays of p53 function gave apparently wild type results on peripheral blood lymphocytes from this individual. These results led us to carry out more detailed functional tests on the mutant protein. The mutant allele was expressed either at very low levels or not at all in phytohaemagglutinin stimulated lymphocytes. Further, the mutant protein was completely non-functional in terms of its ability to transactivate a series of p53-responsive genes (p21(WAF1), bax, PIG3), to transrepress a target gene and to inhibit colony growth in transfected Saos-2 cells. However, surprisingly, data from irradiated peripheral blood lymphocytes and transfected Saos-2 cells, suggested that this truncated, mutant protein retains significant ability to induce apoptosis. PMID- 12085208 TI - A novel germline mutation of PTEN associated with brain tumours of multiple lineages. AB - We have identified a novel germline mutation in the PTEN tumour suppressor gene. The mutation was identified in a patient with a glioma, and turned out to be a heterozygous germline mutation of PTEN (Arg234Gln), without loss of heterozygosity in tumour DNA. The biological consequences of this germline mutation were investigated by means of transfection studies of the mutant PTEN molecule compared to wild-type PTEN. In contrast to the wild-type molecule, the mutant PTEN protein is not capable of inducing apoptosis, induces increased cell proliferation and leads to high constitutive PKB/Akt activation, which cannot be increased anymore by stimulation with insulin. The reported patient, in addition to glioma, had suffered from benign meningioma in the past but did not show any clinical signs of Cowden disease or other hereditary diseases typically associated with PTEN germline mutations. The functional consequences of the mutation in transfection studies are consistent with high proliferative activity. Together, these findings suggest that the Arg234Gln missense mutation in PTEN has oncogenic properties and predisposes to brain tumours of multiple lineages. PMID- 12085206 TI - Intra-tumoural microvessel density in human solid tumours. AB - Over the last decade assessment of angiogenesis has emerged as a potentially useful biological prognostic and predictive factor in human solid tumours. With the development of highly specific endothelial markers that can be assessed in histological archival specimens, several quantitative studies have been performed in various solid tumours. The majority of published studies have shown a positive correlation between intra-tumoural microvessel density, a measure of tumour angiogenesis, and prognosis in solid tumours. A minority of studies have not demonstrated an association and this may be attributed to significant differences in the methodologies employed for sample selection, immunostaining techniques, vessel counting and statistical analysis, although a number of biological differences may account for the discrepancy. In this review we evaluate the quantification of angiogenesis by immunohistochemistry, the relationship between tumour vascularity and metastasis, and the clinicopathological studies correlating intra-tumoral microvessel density with prognosis and response to anti cancer therapy. In view of the extensive nature of this retrospective body of data, comparative studies are needed to identify the optimum technique and endothelial antigens (activated or pan-endothelial antigens) but subsequently prospective studies that allocate treatment on the basis of microvessel density are required. PMID- 12085210 TI - Anti-angiogenic action of hyperthermia by suppressing gene expression and production of tumour-derived vascular endothelial growth factor in vivo and in vitro. AB - Vascular endothelial growth factor is an important angiogenic factor for tumour progression because it increases endothelial-cell proliferation and remodels extracellular matrix in blood vessels. We demonstrated that hyperthermia at 42 degrees C, termed heat shock, suppressed the gene expression and production of vascular endothelial growth factor in human fibrosarcoma HT-1080 cells and inhibited its in vitro angiogenic action on human umbilical vein endothelial cells. The gene expression of alternative splicing variants for vascular endothelial growth factor, VEGF121, VEGF165 and VEGF189, was constitutively detected in HT-1080 cells, but the VEGF189 transcript was less abundant than VEGF121 and VEGF165. When HT-1080 cells were treated with heat shock at 42 degrees C for 4 h and then maintained at 37 degrees C for another 24 h, the gene expression of all vascular endothelial growth factor variants was suppressed. In addition, HT-1080 cells were found to produce abundant VEGF165, but much less VEGF121, both of which were inhibited by heat shock. Furthermore, the level of vascular endothelial growth factor in sera from six cancer patients was significantly diminished 2-3 weeks after completion of whole-body hyperthermia at 42 degrees C (49.9+/-36.5 pg x ml(-1), P<0.01) as compared with that prior to the treatment (177.0+/-77.5 pg x ml(-1)). On the other hand, HT-1080 cell-conditioned medium showed vascular endothelial growth factor-dependent cell proliferative activity and the augmentation of pro-matrix metalloproteinase-1 production in human umbilical vein endothelial cells. The augmentation of endothelial-cell proliferation and pro-matrix metalloproteinase-1 production was poor when human umbilical vein endothelial cells were treated with conditioned medium from heat shocked HT-1080 cells. These results suggest that hyperthermia acts as an anti angiogenic strategy by suppressing the expression of tumour-derived vascular endothelial growth factor production and thereby inhibiting endothelial-cell proliferation and extracellular matrix remodelling in blood vessels. PMID- 12085211 TI - A novel combretastatin A-4 derivative, AC7700, strongly stanches tumour blood flow and inhibits growth of tumours developing in various tissues and organs. AB - In a previous study, we used subcutaneous LY80 tumours (a subline of Yoshida sarcoma), Sato lung carcinoma, and methylcholanthrene-induced primary tumours, to demonstrate that a novel water-soluble combretastatin A-4 derivative, AC7700, abruptly and irreversibly stopped tumour blood flow. As a result of this interrupted supply of nutrients, extensive necrosis was induced within the tumour. In the present study, we investigated whether AC7700 acts in the same way against solid tumours growing in the liver, stomach, kidney, muscle, and lymph nodes. Tumour blood flow and the change in tumour blood flow induced by AC7700 were measured by the hydrogen clearance method. In a model of cancer chemotherapy against metastases, LY80 cells (2x10(6)) were injected into the lateral tail vein, and AC7700 at 10 mg x kg(-1) was injected i.v. five times at intervals of 2 days, starting on day 7 after tumour cell injection. The number and size of tumours were compared with those in the control group. The change in tumour blood flow and the therapeutic effect of AC7700 on microtumours were observed directly by using Sato lung carcinoma implanted in a rat transparent chamber. AC7700 caused a marked decrease in the tumour blood flow of all LY80 tumours developing in various tissues and organs and growth of all tumours including lymph node metastases and microtumours was inhibited. In every tumour, tumour blood flow began to decrease immediately after AC7700 administration and reached a minimum at approximately 30 min after injection. In many tumour capillaries, blood flow completely stopped within 3 min after AC7700 administration. These results demonstrate that AC7700 is effective for tumours growing in various tissues and organs and for metastases. We conclude that tumour blood flow stanching induced by AC7700 may become an effective therapeutic strategy for all cancers, including refractory cancers because the therapeutic effect is independent of tumour site and specific type of cancer. PMID- 12085212 TI - Dietary restriction reduces angiogenesis and growth in an orthotopic mouse brain tumour model. AB - Diet and lifestyle produce major effects on tumour incidence, prevalence, and natural history. Moderate dietary restriction has long been recognised as a natural therapy that improves health, promotes longevity, and reduces both the incidence and growth of many tumour types. Dietary restriction differs from fasting or starvation by reducing total food and caloric intake without causing nutritional deficiencies. No prior studies have evaluated the responsiveness of malignant brain cancer to dietary restriction. We found that a moderate dietary restriction of 30-40% significantly inhibited the intracerebral growth of the CT 2A syngeneic malignant mouse astrocytoma by almost 80%. The total dietary intake for the ad libitum control group (n=9) and the dietary restriction experimental group (n=10) was about 20 and 13 Kcal x day(-1), respectively. Overall health and vitality was better in the dietary restriction-fed mice than in the ad libitum fed mice. Tumour microvessel density (Factor VIII immunostaining) was two-fold less in the dietary restriction mice than in the ad libitum mice, whereas the tumour apoptotic index (TUNEL assay) was three-fold greater in the dietary restriction mice than in the ad libitum mice. CT-2A tumour cell-induced vascularity was also less in the dietary restriction mice than in the ad libitum mice in the in vivo Matrigel plug assay. These findings indicate that dietary restriction inhibited CT-2A growth by reducing angiogenesis and by enhancing apoptosis. Dietary restriction may shift the tumour microenvironment from a proangiogenic to an antiangiogenic state through multiple effects on the tumour cells and the tumour-associated host cells. Our data suggest that moderate dietary restriction may be an effective antiangiogenic therapy for recurrent malignant brain cancers. PMID- 12085213 TI - Orthogonal polarisation spectral imaging as a new tool for the assessment of antivascular tumour treatment in vivo: a validation study. AB - Tumour angiogenesis plays a key role in tumour growth, formation of metastasis, detection and treatment of malignant tumours. Recent investigations provided increasing evidence that quantitative analysis of tumour angiogenesis is an indispensable prerequisite for developing novel treatment strategies such as anti angiogenic and antivascular treatment options. Therefore, it was our aim to establish and validate a new and versatile imaging technique, that is orthogonal polarisation spectral imaging, allowing for non-invasive quantitative imaging of tumour angiogenesis in vivo. Experiments were performed in amelanotic melanoma A MEL 3 implanted in a transparent dorsal skinfold chamber of the hamster. Starting at day 0 after tumour cell implantation, animals were treated daily with the anti angiogenic compound SU5416 (25 mg kg x bw(-1)) or vehicle (control) only. Functional vessel density, diameter of microvessels and red blood cell velocity were visualised by both orthogonal polarisation spectral imaging and fluorescence microscopy and analysed using a digital image system. The morphological and functional properties of the tumour microvasculature could be clearly identified by orthogonal polarisation spectral imaging. Data for functional vessel density correlated excellently with data obtained by fluorescence microscopy (y=0.99x+0.48, r2=0.97, R(S)=0.98, precision: 8.22 cm(-1) and bias: -0.32 cm( 1)). Correlation parameters for diameter of microvessels and red blood cell velocity were similar (r2=0.97, R(S)=0.99 and r2=0.93, R(S)=0.94 for diameter of microvessels and red blood cell velocity, respectively). Treatment with SU5416 reduced tumour angiogenesis. At day 3 and 6 after tumour cell implantation, respectively, functional vessel density was 4.8+/-2.1 and 87.2+/-10.2 cm(-1) compared to values of control animals of 66.6+/-10.1 and 147.4+/-13.2 cm(-1), respectively. In addition to the inhibition of tumour angiogenesis, tumour growth and the development of metastasis was strongly reduced in SU5416 treated animals. This new approach enables non-invasive, repeated and quantitative assessment of tumour vascular network and the effects of antiangiogenic treatment on tumour vasculature in vivo. Thus, quantification of tumour angiogenesis can be used to more accurately classify and monitor tumour biologic characteristics, and to explore aggressiveness of tumours. PMID- 12085214 TI - Development of an in-vitro model system to investigate the mechanism of muscle protein catabolism induced by proteolysis-inducing factor. AB - The mechanism of muscle protein catabolism induced by proteolysis-inducing factor, produced by cachexia-inducing murine and human tumours has been studied in vitro using C(2)C(12) myoblasts and myotubes. In both myoblasts and myotubes protein degradation was enhanced by proteolysis-inducing factor after 24 h incubation. In myoblasts this followed a bell-shaped dose-response curve with maximal effects at a proteolysis-inducing factor concentration between 2 and 4 nM, while in myotubes increased protein degradation was seen at all concentrations of proteolysis-inducing factor up to 10 nM, again with a maximum of 4 nM proteolysis-inducing factor. Protein degradation induced by proteolysis inducing factor was completely attenuated in the presence of cycloheximide (1 microM), suggesting a requirement for new protein synthesis. In both myoblasts and myotubes protein degradation was accompanied by an increased expression of the alpha-type subunits of the 20S proteasome as well as functional activity of the proteasome, as determined by the 'chymotrypsin-like' enzyme activity. There was also an increased expression of the 19S regulatory complex as well as the ubiquitin-conjugating enzyme (E2(14k)), and in myotubes a decrease in myosin expression was seen with increasing concentrations of proteolysis-inducing factor. These results show that proteolysis-inducing factor co-ordinately upregulates both ubiquitin conjugation and proteasome activity in both myoblasts and myotubes and may play an important role in the muscle wasting seen in cancer cachexia. PMID- 12085215 TI - Anti-tumour activity and toxicity of the new prodrug 9-aminocamptothecin glucuronide (9ACG) in mice. AB - Cancer chemotherapy is limited by the modest therapeutic index of most antineoplastic drugs. Some glucuronide prodrugs may display selective anti-tumour activity against tumours that accumulate beta-glucuronidase. We examined the toxicity and anti-tumour activity of 9-aminocamptothecin glucuronide, a new glucuronide prodrug of 9-aminocamptothecin, to evaluate its potential clinical utility. 9-aminocamptothecin glucuronide was 25-60 times less toxic than 9 aminocamptothecin to five human cancer cell lines. Beta-glucuronidase activated 9 aminocamptothecin glucuronide to produce similar cell killing as 9 aminocamptothecin or topotecan. The in vivo toxicity of 9-aminocamptothecin glucuronide in BALB/c mice was dose-, route-, sex- and age-dependent. 9 aminocamptothecin glucuronide was significantly less toxic to female than to male mice but the difference decreased with age. 9-aminocamptothecin glucuronide and 9 aminocamptothecin produced similar inhibition (approximately 80%) of LS174T human colorectal carcinoma tumours. 9-aminocamptothecin glucuronide cured a high percentage of CL1-5 human lung cancer xenografts with efficacy that was similar to or greater than 9-aminocamptothecin, irinotecan and topotecan. The potent anti tumour activity of 9-aminocamptothecin glucuronide suggests that this prodrug should be further evaluated for cancer treatment. PMID- 12085216 TI - Comparison of IgG diffusion and extracellular matrix composition in rhabdomyosarcomas grown in mice versus in vitro as spheroids reveals the role of host stromal cells. AB - The tumour extracellular matrix acts as a barrier to the delivery of therapeutic agents. To test the hypothesis that extracellular matrix composition governs the penetration rate of macromolecules in tumour tissue, we measured the diffusion coefficient of nonspecific IgG in three rhabdomyosarcoma subclones growing as multicellular spheroids in vitro or as subcutaneous tumours in dorsal windows in vivo. In subcutaneous tumours, the diffusion coefficient decreased with increasing content of collagen and sulphated glycosaminoglycans. When grown as multicellular spheroids, no differences in either extracellular matrix composition or diffusion coefficient were found. Comparison of in vitro vs in vivo results suggests an over-riding role of host stromal cells in extracellular matrix production subjected to modulation by tumour cells. Penetration of therapeutic macromolecules through tumour extracellular matrix might thus be largely determined by the host organ. Hence, caution must be exercised in extrapolating drug penetrability from spheroids and multilayer cellular sandwiches consisting of only tumour cells to tumours in vivo. PMID- 12085217 TI - Flavonoids uptake and their effect on cell cycle of human colon adenocarcinoma cells (Caco2). AB - Green tea, mainly through its constituents epigallocatechin gallate, epigallocatechin, epicatechin gallate and epicatechin, has demonstrated anticarcinogenic activity in several animal models, including those for skin, lung and gastro-intestinal tract cancer, although less is known about colorectal cancer. Quercetin, the major flavonoid present in vegetables and fruit, exerts potential anticarcinogenic effects in animal models and cell cultures, but less is known about quercetin glucosides. The objectives of this study were to investigate (i) the antioxidant activity of the phenolic compounds epicatechin, epigallocatechin gallate, gallic acid and quercetin-3-glucoside; (ii) the cytotoxicity of different concentrations of epicatechin, epigallocatechin gallate, and gallic acid; (iii) the cellular uptake of epicatechin, epigallocatechin gallate, gallic acid and quercetin-3-glucoside and (iv) their effect on the cell cycle. Human colon adenocarcinoma cells were used as experimental model. The results of this study indicate that all dietary flavonoids studied (epicatechin, epigallocatechin gallate, gallic acid and quercetin-3-glucoside) show a significant antioxidant effect in a chemical model system, but only epigallocatechin gallate or gallic acid are able to interfere with the cell cycle in Caco2 cell lines. These data suggest that the antioxidant activity of flavonoids is not related to the inhibition of cellular growth. From a structural point of view, the galloyl moiety appears to be required for both the antioxidant and the antiproliferative effects. PMID- 12085219 TI - Treatment of cancer with cryochemotherapy. AB - Cryosurgery employs freezing to destroy solid tumours. However, frozen cells can survive and cause cancer recurrence. Bleomycin, an anticancer drug with a huge intrinsic cytotoxicity is normally not very effective because it is nonpermeant. We report that freezing facilitates bleomycin penetration into cells making it toxic to cryosurgery surviving cells at concentrations that are non-toxic systemically. PMID- 12085218 TI - In vitro and in vivo activity and cross resistance profiles of novel ruthenium (II) organometallic arene complexes in human ovarian cancer. AB - Ruthenium complexes offer the potential of reduced toxicity, a novel mechanism of action, non-cross resistance and a different spectrum of activity compared to platinum containing compounds. Thirteen novel ruthenium(II) organometallic arene complexes have been evaluated for activity (in vitro and in vivo) in models of human ovarian cancer, and cross-resistance profiles established in cisplatin and multi-drug-resistant variants. A broad range of IC50 values was obtained (0.5 to >100 microM) in A2780 parental cells with two compounds (RM175 and HC29) equipotent to carboplatin (6 microM), and the most active compound (HC11) equipotent to cisplatin (0.6 microM). Stable bi-dentate chelating ligands (ethylenediamine), a more hydrophobic arene ligand (tetrahydroanthracene) and a single ligand exchange centre (chloride) were associated with increased activity. None of the six active ruthenium(II) compounds were cross-resistant in the A2780cis cell line, demonstrated to be 10-fold resistant to cisplatin/carboplatin by a mechanism involving, at least in part, silencing of MLH1 protein expression via methylation. Varying degrees of cross-resistance were observed in the P-170 glycoprotein overexpressing multi-drug-resistant cell line 2780AD that could be reversed by co-treatment with verapamil. In vivo activity was established with RM175 in the A2780 xenograft together with non-cross-resistance in the A2780cis xenograft and a lack of activity in the 2780AD xenograft. High activity coupled to non cross-resistance in cisplatin resistant models merit further development of this novel group of anticancer compounds. PMID- 12085220 TI - Apoptosis in human tumours. PMID- 12085225 TI - Resistance to fludarabine-induced apoptosis in Epstein-Barr virus infected B cells. AB - The Epstein-Barr virus (EBV) transforms B cells in part by inhibiting the cellular apoptotic programme. This is also observed when Burkitt lymphoma cell lines are infected with EBV. Induction of apoptosis is one of the mechanisms by which fludarabine inhibits the growth of cells with low proliferative capacity. This compound can also inhibit several other mechanisms in the cell, including inhibition of the synthesis of factors such as STAT1. To analyse the relationship between EBV status, fludarabine-induced apoptosis, and transcription factors we studied the EBV-negative Burkitt lymphoma cell line BL2, its EBV-infected counterpart BL2.B95.8 and the EBV-transformed cell line PRI. The BL2 cell line was found to be very sensitive to fludarabine. The BL2.B95.8 and PRI cells were both resistant but the latter to a lesser extent. In the PRI cells fludarabine activated p53, but not in the BL2.B95.8 cells in which the p53 pathway is inactivated. We observed that this inactivation results in part from the lack of expression of the MDM2 inhibitor p14ARF. Conversely, there was a substantial constitutive activation of STAT1, and not of the other STATs, in the BL2.B95.8 cells and a modest one in the PRI cells. Furthermore, expression of STAT1 was significantly reduced by fludarabine treatment in the PRI cells, but not in the BL2.BL95.8 cells. Finally, the expression of p21WAF1/CIP1 was detected only in the BL2.B95.8 and PRI cells. This protein, known to play a role in cell survival, may therefore be involved in the resistance of the BL2.B95.8 cells to fludarabine. PMID- 12085221 TI - Granisetron vs ondansetron: is it a question of duration of 5-HT3 receptor blockade? PMID- 12085226 TI - Lack of functional pRb results in attenuated recovery of mRNA synthesis and increased apoptosis following UV radiation in human breast cancer cells. AB - Lack of functional pRb results in attenuated recovery of mRNA synthesis and increased apoptosis following UV radiation in human breast cancer cells. We have previously demonstrated that a human breast cancer cell line, MDA-MB-468, which lacks the retinoblastoma protein (pRb), is particularly sensitive to low doses of ultraviolet (UV) radiation. These cells are 15-20-fold more sensitive to UV radiation than cells with wild-type pRb. In order to understand the mechanisms of the high apoptotic response of MDA-MB-468 cells to UV radiation, we examined the effects of UV on these cells with regards to both membrane-mediated events and DNA damage. We found that MDA-MB-468 cells were resistant to all ligand-induced death receptor signaling. In addition, although UV activated caspase 8 in MDA-MB 468 cells, a peptide inhibitor of caspase 8 failed to inhibit UV-induced apoptosis. We then tested the possibility that nuclear events mediated the enhanced sensitivity to UV-induced apoptosis in these cells. Unlike UV-resistant cells, MDA-MB-468 cells were unable to recover mRNA synthesis after 5 J/m2 UVC. We also found that the pRb-null DU-145 cells similarly had attenuated recovery of mRNA synthesis after UV radiation. In UV-resistant cells with wild-type pRb, the inactivation of pRb with HPV-16 E7 resulted in significant inhibition in their ability to recover mRNA synthesis and increased levels of apoptosis following UV radiation. Furthermore, pRb-null cells were deficient in repair of UV radiation induced DNA damage. These data suggest that the sensitivity of MDA-MB-468 cells to UV radiation is due to defects in repair of DNA damage and recovery of mRNA synthesis rather than to membrane death receptor pathways. Inactivation of pRb may contribute to an increased sensitivity to UV radiation by attenuating repair of DNA lesions and recovery of mRNA synthesis following UV radiation. PMID- 12085227 TI - Constitutive activation of NF-kappaB in Ki-ras-transformed prostate epithelial cells. AB - The signaling pathway responsible for the activation of nuclear factor-kappaB (NF kappaB) by oncogenic forms of Ras remains unclear. Both, the transactivation and DNA binding activities of NF-kappaB, were increased in 267B1 human prostate epithelial cells transformed by viral Kirsten-ras (267B1/Ki-ras cells) compared with those in the parental cells. This increased NF-kappaB activity was attributed to a heterodimeric complex of p50 and p65 subunits. Although the abundance of the inhibitor protein IkappaBbeta was higher in 267B1/Ki-ras cells than in 267B1 cells, an electrophoretic mobility-shift assay suggested that IkappaBalpha is responsible for the activation of NF-kappaB in the former cells. Consistent with this notion, the phosphorylation of IkappaBalpha appeared increased in 267B1/Ki-ras cells, and the proteasome inhibitor I abolished the constitutive activation of NF-kappaB in these cells. The expression of dominant negative mutants of either NIK (NF-kappaB-inducing kinase) or IKKbeta (IkappaB kinase beta) inhibited the activity of NF-kappaB in 267B1/Ki-ras cells. Furthermore, chemical inhibitors specific for Ras activation, sulindac sulfide and farnesytranferase inhibitor I, markedly reduced IkappaBalpha phosphorylation and NF-kappaB activation in the Ki-ras-transformed cells while transfection of these cells with NIK or IKKbeta counteracted the inhibitory effect on NF-kappaB activation. These results suggest that oncogenic Ki-Ras induces transactivation of NF-kappaB through the NIK-IKKbeta-IkappaBalpha pathway. PMID- 12085228 TI - Multiple interacting domains contribute to p14ARF mediated inhibition of MDM2. AB - The small basic protein p14ARF, encoded by one of the alternative transcripts from the human INK4A/ARF locus, interferes with MDM2-mediated ubiquitination of the p53 tumour suppressor protein. The resultant stabilization of p53 leads to increased expression of p53-regulated genes, such as MDM2 itself and the cyclin dependent kinase inhibitor p21(CIP1). Here we relate physical interactions between p14ARF and MDM2, as determined using synthetic peptides and systematic deletions of p14ARF, with consequential effects on p53 stabilization and transcriptional activity. The data imply that the amino terminal half of p14ARF, encoded by the alternative first exon (exon 1beta) contacts MDM2 through multiple domains that can independently impede MDM2-mediated degradation of p53, provided that they are localized in the cell nucleus. As well as identifying previously unrecognized functional domains, our findings offer an explanation for the relative paucity of missense mutations in exon 1beta in human tumours. PMID- 12085229 TI - Analysing c-kit internalization using a functional c-kit-EGFP chimera containing the fluorochrome within the extracellular domain. AB - In order to investigate activation and internalization of c-kit we created a functional c-kit-EGFP chimera by inserting EYFP (enhanced yellow fluorescent protein) within the extracellular domain of c-kit immediately downstream of the signal sequence, SS-EYFP-kit. This location was chosen because the C-terminal fusion of EGFP to c-kit unexpectedly caused constitutive activation of the c-kit tyrosine kinase. As analysed in fixed cells and by real time imaging in vivo, SCF induced activation led to internalization of the fusion construct and translocation to punctate structures resembling vesicles. Analysis of the internalization process by time lapse imaging revealed high mobility and discontinuous movement of these vesicles and their predominantly radial tracks. Two subsets of vesicles were observed: Traffic of the majority of vesicles was directed from the periphery to the center of the cell and most likely represents the internalization of activated receptor molecules via the endosomal pathway. However, some vesicular structures were observed to move towards the periphery of the cell and probably contain newly synthesized protein to replace internalized receptor molecules. The calculated velocity of moving vesicles ranged from 0.05 to 0.2 microm per se. Vesicle formation upon SCF induced dimerization of the receptor was strictly dependent on kinase activity of c-kit. Treatment of cells with phenylarsine oxide, an agent blocking receptor internalization, prior to SCF stimulation resulted in abrogation of the translocation of the chimera to vesicles whereas accumulation of vesicles was observed when cells were treated with proteasome inhibitors. Cholesterol depletion of the cell membrane by methyl beta-cyclodextrin resulted in dose dependent reduction of receptor internalization indicating that c-kit may be present in lipid rafts or that intact lipid rafts are required for efficient internalization of the receptor. Using the induction of vesicular structures as a sign of efficient internalization of the receptor analysis of mutant c-kit constructs deficient either in activation of PI3-Kinase or Src revealed that internalization of c-kit is dependent on recruitment of Src but not PI3-Kinase. PMID- 12085230 TI - Cre-loxP chromosome engineering of a targeted deletion in the mouse corresponding to the 3p21.3 region of homozygous loss in human tumours. AB - Chromosomal deletions are a common feature of epithelial tumours and when further defined by homozygous deletions, are often the location of tumour suppressor genes. Deletions within the short arm of chromosome 3 occur very frequently in human carcinomas: a minimal region of loss at 3p21.3 (the Luca) region has been defined by overlapping homozygous deletions in lung and breast cancer cell lines. Using a rapid strategy for Cre-loxP chromosome engineering, a deletion of approximately 370 kb was created in the mouse germline corresponding to the deleted region at 3p21.3. The deletion when homozygous is embryonic lethal. Heterozygotes develop normally despite being haplo-insufficient for twelve genes including the candidate tumour suppressor gene Rassf1. Because damage to 3p21.3 often occurs very early in the sequence of genetic changes that lead to malignancy, particularly in lung and breast cancer, further genetic damage to these mice will provide the opportunity to model multi-step tumorigenesis of these tumours. PMID- 12085231 TI - Induction of apoptosis by the new anticancer drug XK469 in human ovarian cancer cell lines. AB - XK469, a synthetic quinoxaline phenoxypropionic acid derivative, has been found to have selective activity against a broad panel of solid tumors including several drug-resistant cell lines and has been approved for phase I clinical evaluation. Recent studies suggested that XK469 is a selective topoisomerase IIbeta inhibitor, but the mechanism of XK469-induced cell death remains unknown. Here we investigate the ability of XK469 to induce apoptosis of human cancer cells. In the human ovarian cancer cell line PA1, XK469 caused the release of cytochrome c, activation of caspases including caspases 9, 7 and 3, cleavage of PARP, and subsequently cell death. Moreover, Bcl2 and Bax were cleaved in XK469 treated cells. PA1 cells expressing the dominant negative-caspase 9 were less sensitive to XK469. Importantly, in these PA1 cells expressing DN-casp 9, the activation of caspases including caspases 3, 7 and 9, and cleavage of Bax and Bcl2 were inhibited, suggesting that the activation of the mitochondrial pathway is required for XK469-induced anticancer activity. These results indicate that the induction of apoptosis by XK469 may account for its anti-tumor activity and such activity is required for the activation of the mitochondrial pathway. Thus, our study defines a possible mechanism, at least in part, underlying XK469 induced anti-cancer activity. PMID- 12085232 TI - Differential regulation of Rb family proteins and prohibitin during camptothecin induced apoptosis. AB - Prohibitin, a potential tumor suppressor, is known to induce growth suppression and repress E2F-mediated transcription. These growth regulatory functions of prohibitin require a physical interaction with the Rb protein. We now find that prohibitin protects cells from apoptosis mediated by camptothecin, a topoisomerase I inhibitor. Camptothecin treatment of Ramos B cells leads to the degradation of Rb protein and phosphorylation of its family members, p107 and p130. This correlates with an increase in the levels of cyclin E as well as the kinase activity associated with it. Inactivation of Rb leads to the dissociation and release of free E2F. We find also that E2F activity is induced upon camptothecin treatment, but this increase is absent in prohibitin overexpressing cells. It thus appears that prohibitin may be inhibiting apoptosis by downregulating E2F activity when Rb family members are inactive. PMID- 12085233 TI - N-Myc and Bcl-2 coexpression induces MMP-2 secretion and activation in human neuroblastoma cells. AB - Neuroblastoma is a peripheral nervous system tumor that accounts for 8-10% of all solid childhood tumors. N-Myc is the most reliable prognostic indicator for neuroblastoma. Bcl-2 is detected in 40-60% of primary neuroblastoma tumors and demonstrates anti-apoptotic action by conferring resistance to chemotherapy and radiation treatment. In neuroblastoma cell lines, the coexpression of N-Myc and Bcl-2 leads to increased tumorigenic properties. Matrix metalloproteinases (MMPs) are endopeptidases that degrade a wide range of basement membrane components, a process important for tumor invasion. This study investigates the effect of N-Myc and Bcl-2 on MMP expression and activation. MMP-2 expression and secretion are increased in SHEP neuroblastoma cells expressing Bcl-2 alone (SHEP/Bcl-2 cells) or both N-Myc and Bcl-2 (SHEP/N-Myc/Bcl-2 cells). MMP-2 activity is increased in the SHEP/N-Myc/Bcl-2 cells yet remains unchanged in SHEP/Bcl-2 cells. TIMP-2 expression is high in SHEP/Bcl-2 cells, which likely inhibits MMP-2 activity, and absent in SHEP/N-Myc/Bcl-2 cells, allowing MMP-2 activity. Invasion is increased in SHEP/N-Myc/Bcl-2 cells and prevented by the use of a pharmacologic MMP-2 inhibitor. These data imply that N-Myc and Bcl-2 cooperate to increase the expression, secretion, and activation of MMP-2, which likely leads to a more tumorigenic phenotype due to increased MMP-2 mediated invasion. PMID- 12085234 TI - Inhibition of human lung cancer growth following adenovirus-mediated mda-7 gene expression in vivo. AB - Overexpression of the melanoma differentiation associated gene-7 (mda-7) in vitro results in suppression of lung cancer cell proliferation. However, the ability of MDA-7 to suppress lung cancer in vivo has not been previously demonstrated. In this study, we investigated the possibility of inducing overexpression of the mda 7 gene in human non-small cell lung carcinoma cells in vivo and its effects on tumor growth. Adenovirus-mediated overexpression of MDA-7 in p53-wild-type A549 and p53-null H1299 subcutaneous tumors resulted in significant tumor growth inhibition through induction of apoptosis. In addition, decreased CD31/PECAM expression and upregulation of APO2/TRAIL were observed in tumors expressing MDA 7. In vivo studies correlated well with in vitro inhibition of lung tumor cell proliferation and endothelial cell differentiation mediated by Ad-mda7. These data demonstrate that Ad-mda7 functions as a multi-modality anti-cancer agent, possessing both, pro-apoptotic and anti-angiogenic properties. We demonstrate for the first time the potential therapeutic effects of Ad-mda7 in human lung cancer. PMID- 12085235 TI - Distinct roles for phosphoinositide 3-kinase, mitogen-activated protein kinase and p38 MAPK in mediating cell cycle progression of breast cancer cells. AB - Addition of the ErbB-ligand, Heregulinbeta1 (HRG), to breast tumour-derived T47D cells promotes D-cyclin expression, p21(cip1) synthesis, cyclin-dependent kinase (CDK) activation through re-distribution of p27(kip1) and DNA synthesis. In contrast EGF has no effect on T47D cell cycle progression. By comparing these two ligands and the use of specific inhibitors for phosphatidylinositol-3 kinase (PI3K), mitogen-activated protein kinase (MAPK) and p38MAPK, we have identified several molecular mechanisms required for ErbB receptor-mediated proliferation. The PI3K, MAPK and p38MAPK pathways each displayed distinct activation profiles in response to either HRG or EGF, with obvious differences in both the intensity and duration of signal output. Through inhibition of each of these pathways it is apparent that each pathway is necessary, yet insufficient alone, to stimulate proliferation. Each pathway regulates distinct subsets of essential cell cycle regulators and integration of these signal networks is required for the timely expression of these components, which culminates in cell cycle progression. Significantly, the mechanisms controlling ligand-stimulated proliferation through ErbB2 are strikingly similar to the mechanisms through which overexpressed, constitutively activated, ErbB2 orchestrates uncontrolled proliferation in cancer cells. This suggests that downstream effectors of ErbB receptors represent good therapeutic targets for breast cancer. PMID- 12085236 TI - Immortalization and transformation of primary human airway epithelial cells by gene transfer. AB - One critical step in the development of a cancerous cell is its acquisition of an unlimited replicative lifespan, the process termed immortalization. Experimental model systems designed to study cellular transformation ex vivo have relied to date on the in vitro selection of a subpopulation of cells that have become immortalized through treatment with chemical or physical mutagens and the selection of rare clonal variants. In this study, we describe the direct immortalization of primary human airway epithelial cells through the successive introduction of the Simian Virus 40 Early Region and the telomerase catalytic subunit hTERT. Cells immortalized in this way are now responsive to malignant transformation by an introduced H-ras or K-ras oncogene. These immortalized human airway epithelial cells, which have been created through the stepwise introduction of genetic alterations, provide a novel experimental model system with which to study further the biology of the airway epithelial cell and to dissect the molecular basis of lung cancer pathogenesis. PMID- 12085237 TI - Expression profiling of microdissected pancreatic adenocarcinomas. AB - Pancreatic ductal adenocarcinoma is characterized by a paucity of neoplastic cells embedded in a densely desmoplastic stroma. Therefore, laser capture microdissection was performed to obtain homogeneous populations of normal and neoplastic ductal cells. These were subjected to a comparative study of gene expression utilizing human cDNA arrays. A variety of dysregulated genes were identified, comprising cell cycle and growth regulators, invasion regulators, signalling and developmental molecules. In addition to genes already found to be overexpressed in pancreatic cancer, such as TIMP1, MMP7, CD59, rhoC and NDKA, we present evidence to implicate genes which have not previously been reported in this tumour type. These include the overexpressed genes ABL2, Notch4 and SOD1, as well as XRCC1, a DNA repair gene whose transcript was found downregulated. Quantitative real-time RT-PCR (QRT-PCR) was employed to confirm differential expression of ABL2, Notch4 and SOD1 and immunohistochemical analysis was used to verify decreased protein expression of XRCC1 using a custom-built pancreatic tissue array. Combining microarray-derived gene expression profiles of pure pancreatic cell populations, QRT-PCR and pancreas-specific tissue arrays therefore proved to be highly informative in elucidating the molecular pathology of this highly malignant tumour type. PMID- 12085238 TI - Role of integrin alphaVbeta3 in the production of recombinant adenoviruses in HEK 293 cells. AB - Adenoviral infection is initiated by attachment of adenoviral fiber proteins to the CAR protein and subsequent internalization aided by alphaV -containing integrins, eg alphaVbeta3 and alphaVbeta5. To further understand the process of infection and assembly of recombinant adenoviral (rAd) vectors, we examined rAd production in HEK-293 cells and one of its subclones, clone D, isolated from the parental cells for high viral production. By flow cytometry, surface expression of integrin alphaVbeta3 by clone D cells was two-fold higher than by HEK-293 cells. However, clone D cells did not demonstrate greater translational efficiency or number of viral genome DNA copies shortly after rAd infection. Treating cells with inhibitors of integrin alphaVbeta3 reduced rAd production and transfecting HEK-293 cells with integrin alphaVbeta3 cDNAs increased rAd production. Subjecting cells to a sudden reduction in serum (10% to 0.1% FCS) for 5 days, clone D cells maintained 80% viability compared with 40% for HEK-293 cells. Further indication of survival signaling involvement was provided by Western blot analysis demonstrating p38 and p44/42 MAPKs were constitutively phosphorylated in HEK-293 cells. However, for clone D cells, p38 MAPK was phosphorylated only after rAd infection. The role of survival signaling mediated by integrin alphaVbeta3 in rAd production will be discussed. PMID- 12085239 TI - Enhanced engraftment of EPO-transduced human bone marrow stromal cells transplanted in a 3D matrix in non-conditioned NOD/SCID mice. AB - Intravenous infusion of bone marrow stromal cells (BMSCs) has been proposed as a means to support hematopoiesis in bone marrow transplantation or as a vehicle for gene therapy. However, it seems that this route of injection leads to engraftment of a small proportion of BMSCs, possibly because they are unable to cross the endothelial barrier. We have transplanted human BMSCs, ex vivo expanded and transduced with a retrovirus encoding the human erythropoietin gene, either intravenously or subcutaneously with or without a tridimensional scaffold in non conditioned NOD/SCID mice. Efficiency of engraftment was evaluated monitoring the hematocrit levels. Systemic infusion never significantly increased hematocrit levels, whereas subcutaneous transplantation of the same number of cells induced an important increase of the hematocrit (approximately 70%) for at least 2 months. A substantial increase in the length of the response was observed when cells were subcutaneously transplanted in a tridimensional scaffold. To determine whether the transient effect was due to cell loss or to reduction in expression, the cells implanted into a tridimensional scaffold were recovered, expanded in vitro, and re-implanted in a new group of mice. Again the hematocrit levels rose 2 weeks after transplantation ( approximately 70%). These results demonstrate that ex vivo expanded human BMSCs are not quantitatively transplantable by systemic infusion in non-conditioned recipients, whereas the local implantation into a tridimensional scaffold allows long-term engraftment and efficient expression of a foreign gene. PMID- 12085240 TI - Apical barriers to airway epithelial cell gene transfer with amphotropic retroviral vectors. AB - Gene transfer to airway epithelia with amphotropic pseudotyped retroviral vectors is inefficient following apical vector application. To better understand this inefficiency, we localized the expression of Pit2, the amphotropic receptor, in polarized human airway epithelia. Pit2 was expressed on both the apical and basolateral surfaces of the cells, suggesting that factors other than receptor abundance may limit apical gene transfer efficiency. Binding studies performed with radiolabeled amphotropic MuLV suggested that the apically applied virus binds to Pit2. Hypothetical barriers to retroviral gene transfer include the apical glycocalyx and other secreted products of epithelia. In this study, we demonstrated that sialic acid, keratan sulfate and collagen type V are present on the apical surface of well-differentiated human airway epithelia. While enzyme treatment reduced the abundance of these components, the treatment also decreased the transepithelial resistance to approximately 35% of the controls, suggesting that the epithelial integrity was impaired. To attain an airway epithelial culture with a modified apical surface and intact epithelial integrity, we utilized 100 mM 2-deoxy-D-glucose, a glycosylation inhibitor, to prevent the glycocalyx from reforming following enzyme treatment. This approach allowed the resistance, but not the apical glycocalyx to recover. Despite this physical modification of the cell surface, the amphotropic retroviral vector failed to transduce airway epithelia following apical application. These results suggest that factors other than apical receptor abundance and the glycocalyx inhibit amphotropic retroviral gene transfer in human airway epithelia. PMID- 12085241 TI - Gene transduction efficiency in cells of different species by HIV and EIAV vectors. AB - The ability of human immunodeficiency virus (HIV)- and equine infectious anaemia virus (EIAV)-based vectors to transduce cell lines from a range of species was compared. Both vectors carried the vesicular stomatitis virus G (VSV-G) envelope protein and encoded an enhanced green fluorescent protein (eGFP) gene driven by a human cytomegalovirus (CMV) early promoter. Immunostaining for viral core proteins and VSV-G was used to demonstrate that the HIV and EIAV vector preparations contained similar numbers of virus particles. Various cell lines were transduced with these vectors and the transduction efficiency was estimated by measuring eGFP expression. Efficient transduction by both vectors was observed in human, hamster, pig, horse, cat and dog cell lines, although EIAV vector was about 10-fold less efficient in human, hamster and pig cells normalised to the total number of viral particles. This could be partly explained by the lower RNA genome levels per particle for EIAV as measured by real-time RT-PCR. Rodent cells appeared to be transduced inefficiently with both vectors, but when the CMV promoter was substituted with the EF1alpha promoter in the HIV vectors, the expression level increased leading to an increase in the measurable level of transduction. PMID- 12085242 TI - Inducing neutrophil recruitment in the liver of ICAM-1-deficient mice using polyethyleneimine grafted with Pluronic P123 as an organ-specific carrier for transgenic ICAM-1. AB - Coordinated expression of cell adhesion molecules and chemokines on the surface of vascular endothelium is responsible for the homing of immune effector cells to targeted sites. One way to attract non-activated immune cells to targeted organs is to use transgenically expressed adhesion molecules responsible for leukocyte recruitment. We have previously shown that polyethyleneimine (PEI) grafted with non-ionic amphiphilic Pluronic P123 block copolymer (P123PEI) modifies biodistribution of plasmid DNA toward the liver. In the present study, a P123PEI formulated plasmid carrying the gene encoding for the murine ICAM-1 molecule was injected i.v. into transgenic ICAM-1-deficient mice. The RT-PCR analysis of ICAM 1 mRNA expression showed that P123PEI induced a dose-dependent expression of ICAM 1 in the liver. Furthermore, this expression of ICAM-1 induced neutrophil invasion in the liver, while no such invasion was observed in mice injected with formulated control plasmid or naked DNA. These results suggest that P123PEI allows functional transgene expression in the liver following i.v. injection and that ICAM-1 could be used to enhance immune response locally by attracting immune effector cells. PMID- 12085244 TI - Suicide gene transduction sensitizes murine embryonic and human mesenchymal stem cells to ablation on demand-- a fail-safe protection against cellular misbehavior. AB - Stem cells and their progeny constitute a potential resource for replacing damaged tissues or supplying missing functions, but also pose a threat of aberrant behavior, including neoplastic growth or immunopathology. Suicide genes introduced into these cells before transplantation might provide a means of addressing this threat by permitting the ablation of the cells if they subsequently misbehave. Retroviral transduction of the E. coli gpt and herpes thymidine kinase (HSVtk) suicide genes was used to determine the degree to which stem cells could be sensitized to the prodrugs 6-thioxanthine (6TX) and ganciclovir (GCV) respectively, and whether this sensitivity could persist over many cell generations. The ES-E14TG2a murine embryonic stem cell line was rendered sensitive to quantitative ablation at prodrug concentrations well tolerated by untransduced cells (50 microM 6TX, 1 microg/ml GCV). The HSVtk gene also conferred GCV sensitivity on human mesenchymal stem cells and hematopoietic precursors derived from the murine cells, although ablation was not complete. Because ES-E14TG2a cells are deficient in the cellular enzyme HPRT, they are sensitive to hypoxanthine/aminopterin/thymidine (HAT). This property enhanced the persistence of chemosensitivity in gpt-transduced cells by permitting cells that lost 6TX sensitivity to be ablated with HAT. PMID- 12085243 TI - Oxygen-sensitive enzyme-prodrug gene therapy for the eradication of radiation resistant solid tumours. AB - Overwhelming clinical and experimental data demonstrate that tumour hypoxia is associated with aggressive disease and poor treatment outcome as hypoxic cells are refractive to radiotherapy and some forms of chemotherapy. However, hypoxia is rare in physiologically normal tissues representing a tumour-specific condition. To selectively target this therapeutically refractive cell population, we have combined bioreductive chemotherapy with hypoxia-directed gene therapy. We have transfected the human fibrosarcoma cell line, HT1080, with a hypoxia regulated expression vector encoding the human flavoprotein cytochrome c P450 reductase (HRE-P450R). This conferred hypoxia-dependent sensitivity to the alkylating nitroimidazole prodrug RSU1069 in vitro, with a greater than 30-fold increase in oxic/hypoxic cytotoxicity ratio compared with controls. Xenografts of both the HRE-P450R and empty vector transfectants had comparable hypoxic fractions and were refractive to single dose radiotherapy of up to 15 Gy. However, combining a prodrug of RSU1069 with a reduced radiotherapy dose of 10 Gy represents a curative regimen (50% tumour-free survival; day 100) in the HRE P450R xenografts. In complete contrast, 100% mortality was apparent by day 44 in the empty vector control xenografts treated in the same way. Thus, an oxygen sensitive gene-directed enzyme prodrug therapy approach may have utility when incorporated into conventional radiotherapy and/or chemotherapy protocols for loco-regional disease in any tissue where hypoxia is a contra-indication to treatment success. doi:10.1038/sj.gt.3301702 PMID- 12085245 TI - Constitutive and regulated expression of processed insulin following in vivo hepatic gene transfer. AB - To test whether hepatocytes engineered in vivo can serve as surrogate beta cells by similarly secreting mature insulin in a glucose-sensitive manner, we prepared adenoviral vectors encoding wild-type proinsulin (hIns-wt), a modified proinsulin cleavable by the ubiquitously expressed protease furin (hIns-M3), or each of the two beta cell specific pro-insulin convertases PC2 and PC3. Following a detailed in vitro characterization of the proteins produced by our vectors, we infected the liver and, for comparison, the muscle of a chemically induced murine model of type I diabetes. Insulin expression from the transduced tissues was extensively characterized and showed to be constitutive rather than regulated. To obtain regulated expression, we placed expression of hIns-M3 under the control of the dimerizer-inducible transcription system. Hormone secretion from mouse liver was negligible in the absence of the dimerizer drug rapamycin, was inducible in a dose-dependent manner upon its administration, and reversible following drug withdrawal. These data confirm liver as a promising target for in vivo expression of processed insulin. While suggesting that hepatocytes cannot provide authentic glucose-responsive regulation, these results demonstrate that pharmacological regulation is a promising alternative route to the controlled delivery of insulin following hepatic gene transfer. PMID- 12085246 TI - Transcriptional targeting of conditionally replicating adenovirus to dividing endothelial cells. AB - Conditionally replicating adenoviruses (CRADs) are a novel strategy in cancer treatment and clinical trials using CRADs targeted to tumor cells have been reported recently. We hypothesized that it would be possible to construct CRADs targeted to dividing endothelial cells, which are present in the tumor endothelium. We utilized the regulatory elements of Flk-1 and endoglin genes, which have been shown to be highly overexpressed in angiogenic endothelial cells, to construct two CRADs: Ad.Flk-1, which has adenoviral E1A gene under the control of the Flk-1 enhancer/promoter, and Ad.Flk-Endo, which harbors the same Flk-1 enhancer/promoter as Ad.Flk-1, plus it has the adenoviral E1B gene under control of the endoglin promoter. Viral titer measurements by plaque assay showed that in human umbilical vein endothelial cells (HUVECs), both CRADs replicated at levels comparable to that of wild-type adenovirus. In Flk-1 and endoglin negative Hep3B and A549 cells, however, the replication of Ad.Flk-1 and Ad.Flk-Endo was reduced by 30-fold and 600-fold, respectively. Cytotoxicity assays demonstrated that both CRADs killed HUVECs as effectively as wild-type adenovirus and their cytotoxicity in Hep3B and A549 cells was comparable to nonreplicating control adenovirus. Furthermore, there was a striking inhibition (83-91%) of capillary network formation in an in vitro angiogenesis assay when HUVECs were infected with Ad.Flk 1 or Ad.Flk-Endo as compared with the nonreplicating control virus. These results demonstrate that CRADs can be transcriptionally targeted to dividing endothelial cells with high specificity, and that the combined use of Flk-1 and endoglin regulatory elements has a synergistic effect on targeting specificity. This principle may be incorporated into novel therapeutic agents to develop anti angiogenic treatment for cancer. PMID- 12085247 TI - Does training increase the use of more emotionally laden words by nurses when talking with cancer patients? A randomised study. AB - The emotional content of health care professionals-cancer patient communication is often considered as poor and has to be improved by an enhancement of health care professionals empathy. One hundred and fifteen oncology nurses participating in a communication skills training workshop were assessed at three different periods. Nurses randomly allocated to a control group arm (waiting list) were assessed a first time and then 3 and 6 months later. Nurses allocated to the training group were assessed before training workshop, just after and 3 months later. Each nurse completed a 20-min clinical and simulated interview. Each interview was analysed by three content analysis systems: two computer-supported content analysis of emotional words, the Harvard Third Psychosocial Dictionary and the Martindale Regressive Imagery Dictionary and an observer rating system of utterances emotional depth level, the Cancer Research Campaign Workshop Evaluation Manual. The results show that in clinical interviews there is an increased use of emotional words by health care professionals right after having been trained (P=0.056): training group subjects use 4.3 (std: 3.7) emotional words per 1000 used before training workshop, and 7.0 (std: 5.8) right after training workshop and 5.9 (std: 4.3) 3 months later compared to control group subjects which use 4.5 (std: 4.8) emotional words at the first assessment point, 4.3 (std: 4.1) at the second and 4.4 (std: 3.3) at the third. The same trend is noticeable for emotional words used by health care professionals in simulated interviews (P=0.000). The emotional words registry used by health care professionals however remains stable over time in clinical interviews (P=0.141) and is enlarged in simulated interviews (P=0.041). This increased use of emotional words by trained health care professionals facilitates cancer patient emotion words expressions compared to untrained health care professionals especially 3 months after training (P=0.005). This study shows that health care professionals empathy may be improved by communication skills training workshop and that this improvement facilitates cancer patients emotions expression. PMID- 12085248 TI - Outcome after extended follow-up in a prospective study of operable breast cancer: key factors and a prognostic index. AB - In 1990, 215 patients with operable breast cancer were entered into a prospective study of the prognostic significance of five biochemical markers and 15 other factors (pathological/chronological/patient). After a median follow-up of 6.6 years, there were 77 recurrences and 77 deaths (59 breast cancer-related). By univariate analysis, patient outcome related significantly to 13 factors. By multivariate analysis, the most important of nine independent factors were: number of nodes involved, steroid receptors (for oestrogen or progestogen), age, clinical or pathological tumour size and grade. Receptors and grade exerted their influence only in the first 3 years. Progestogen receptors (immunohistochemical) and oestrogen receptors (biochemical) were of similar prognostic significance. The two receptors were correlated (r=+0.50, P=0.001) and displaced each other from the analytical model but some evidence for the additivity of their prognostic values was seen when their levels were discordant. PMID- 12085249 TI - A phase I dose-escalating study of DaunoXome, liposomal daunorubicin, in metastatic breast cancer. AB - The aims of this phase I study were to establish the maximum tolerated dose, safety profile and activity of liposomal daunorubicin, DaunoXome (NeXstar Pharmaceuticals), in the treatment of metastatic breast cancer. DaunoXome was administered intravenously over 2 h in 21 day cycles and doses were increased from 80 to 100, 120 and 150 mg m(2). Sixteen patients were enrolled. A total of 70 cycles of DaunoXome were administered. The maximum tolerated dose was 120 mg m(2), the dose-limiting toxicity being prolonged grade 4 neutropenia or neutropenic pyrexia necessitating dose reductions at 120 and 150 mg m(2). Asymptomatic cardiotoxicity was observed in three patients: grade 1 in one treated with a cumulative dose of 800 mg m(2) and grade 2 in two, one who received a cumulative dose of 960 mg m(2) and the other a cumulative dose of 600 mg m(2) with a previous neoadjuvant doxorubicin chemotherapy of 300 mg m(2). Tumour response was evaluable in 15 patients, of whom two had objective responses, six had stable disease and seven had progressive disease. In conclusion, DaunoXome is associated with mild, manageable toxicities and has anti tumour activity in metastatic breast cancer. The findings support further phase II evaluation of DaunoXome alone and in combination with other standard non anthracycline cytotoxic or novel targeted agents. Although the dose-limiting toxicity for DaunoXome was febrile neutropenia at 120 mg m(2), we would recommend this dose for further evaluation, as the febrile neutropenia occurred after four or more cycles in three of the four episodes seen, was short lived and uncomplicated. PMID- 12085250 TI - Immune changes in patients with advanced breast cancer undergoing chemotherapy with taxanes. AB - Besides cytotoxicity, taxanes induce other biological effects, especially in the immune system. Taxanes have demonstrated immunostimulatory effects against neoplasms, supporting the idea that these agents suppress cancer through several mechanisms and not solely through inhibiting cell division. The purpose of the present study was to evaluate the effect of taxanes (paclitaxel and docetaxel) and investigate their ability in alterating important immunological parameters in breast cancer patients. Thirty women with advanced breast cancer undergoing chemotherapy were randomly assigned into two groups treated with either single agent Paclitaxel or Docetaxel. Sera from patients before the first and after the last treatment cycle and from normal donors were assayed by ELISA for IL-2, IL 1beta, IFN-gamma, GM-CSF, IL-6, TNF-alpha, and PGE2 levels. In these same blood samples, NK and LAK cell activity was tested in the total PBMC population against NK-sensitive K562 tumour targets, respectively, and autologous mixed lymphocyte reaction was tested by (3)H-thymidine proliferation assays. All patients in both groups responded to therapy. Significant differences were observed in the following immune parameters between the control group of healthy blood donors and the pretreatment values of both taxane groups; IL-2, GM-CSF, IFN-gamma levels and NK and LAK cell cytotoxicity were depressed, whereas TNF-alpha and IL-6 levels were raised in breast cancer patients before treatment compared to controls. There were no significant differences between the two treatment groups regarding any of the parameters studied. Both drugs led to increases in MLR values, NK and LAK cell cytotoxicity, and IL-6, GM-CSF, IFN-gamma levels, and decreases for IL 1, TNF, and PGE2 levels. The percentage of these differences was greater for docetaxel in comparison to paclitaxel (P<0.0001). More specifically, docetaxel demonstrated a more pronounced effect on enhancing MLR, NK, LAK activity and IFN gamma, IL-2, IL-6, and GM-CSF levels, as well as caused more potent reduction in IL-1 and TNF-alpha levels when compared to paclitaxel. The present study indicates that patients responded to treatment of advanced breast cancer with single-agent paclitaxel or docetaxel leads to an increase in serum IFN-gamma, IL 2, IL-6, GM-CSF cytokine levels and enhancement of PBMC NK and LAK cell activity, while they both lead to a decrease of acute phase serum cytokine levels of IL-1 and TNF-alpha. Moreover, the effects of docetaxel are in all the above parameters more pronounced than those of paclitaxel. PMID- 12085251 TI - Reduced risk of synovial sarcoma in females: X-chromosome inactivation? AB - Synovial sarcoma shows a characteristic t(X;18) translocation but not the expected female predominance in incidence. We speculate that, among females, one X-chromosome is inactivated and that only the translocation to an active X chromosome leads to development of synovial sarcoma. Population-based cancer registry data from the SEER program support this hypothesis. PMID- 12085252 TI - A prospective cohort study of soy product intake and stomach cancer death. AB - The relationship between intake of soy products and death from stomach cancer was examined in a community-based prospective study of Japanese men and women in Takayama, Japan. Over 7 years of follow-up, 121 deaths from stomach cancer (81 men and 40 women) occurred among 30,304 (13,880 men and 16,424 women) participants who were at least 35 years of age. Diet including the intake of soy products and isoflavones was assessed by a validated semiquantitative food frequency questionnaire at the beginning of the study. In men, the highest compared to the lowest tertile of total soy product intake was significantly inversely associated with death from stomach cancer after controlling for covariates (hazard ratios=0.50; 95% confidence intervals (CIs) 0.26-0.93, P for trend=0.03). Decreased hazard ratios for the highest compared to the lowest tertiles of total soy product intake (hazard ratios=0.49; 95% CI 0.22-1.13) was observed in women, although this association was of marginal significance. These data suggest that soy intake may reduce the risk of death from stomach cancer. PMID- 12085254 TI - Recent trends and future directions for lung cancer mortality in Europe. AB - Lung cancer mortality patterns throughout Europe are very heterogeneous and largely reflect past smoking habits. In order to clarify the changing patterns of lung cancer in Europe we have plotted the overall lung cancer trends among men and women for 20 countries from 1950 up to 1998. Furthermore, using a Bayesian age-period-cohort approach, we have calculated 5 year projections of lung cancer rate up to 2003. Finally, we make some comments on probable future trends by analysing recent trends in adults aged <55 years. Lung cancer mortality rates up to age 75 years portray a general trend of decreasing lung cancer rates among men and increasing lung cancer rates among women. Exceptions to this decrease among men include Hungary where not only are current mortality rates much higher than previously observed in any other country (at 76.7 out of 100,000 in 1998) but they are projected to increase further in the short term. Rates among adults aged <55 years have recently peaked, indicating that overall rates are likely to peak in the next decade. Among women, rapid increases have been observed in Denmark, Netherlands, Hungary, Ireland and UK. Whereas Ireland and UK rates have started to decrease and are projected to continue falling, rates in the other three countries are projected to increase further. Trends in women aged <55 years indicate that rates in Danish women will peak in the next decade, whereas lung cancer rates among Dutch women are likely to continue increasing. Rates in Hungarian women are likely to increase and will surpass the current high rate observed in Denmark. PMID- 12085253 TI - Dietary factors and stomach cancer mortality. AB - The present study examined the relationship between stomach cancer and the low intake of fresh fruit and vegetables and/or a high intake of pickled, preserved or salted foods and frequent use of cooking oil. During 139,390 person-year of follow-up of over 13,000 subjects, 116 died from stomach cancer. Using a Cox proportional hazards-regression analysis of relative risk (RR, 95% CI) controlling for age, sex, smoking and other dietary factors, a significant decline was found with a high consumption of green and yellow vegetables (RR=0.4, 95% CI=0.2-0.9). Reductions of between 40 and 50% were also observed with a high consumption of fresh foods (fruit, cuttle fish, tofu, and potatoes), but these associations were not statistically significant. The risk was significantly increased by the high consumption of processed meat (RR=2.7, 95% CI=1.0-7.4) and by the frequent use of cooking oil (RR=4.0, 95% CI=1.3-11.8). The high consumption of pickled food and traditional soups also increased risk, but not significantly. The findings suggest that a diet high in salt and low in vitamins may be associated with an increase in stomach cancer. PMID- 12085255 TI - Aspirin and lung cancer in women. AB - The association between aspirin use and lung cancer risk in women was examined in a case-control study nested in the New York University Women's Health Study, a large cohort in New York. Case subjects were all the 81 incident lung cancer cases who had provided information about aspirin use at enrollment and during the 1994-1996 follow up. Ten controls per case were randomly selected from among study participants who matched a case by age, menopausal status, and dates of enrollment and follow-up. Relative to no aspirin use, the odds ratio for lung cancer (all histological sub-types combined) among subjects who reported aspirin use three or more times per week for at least 6 months was 0.66 (95% confidence interval 0.34-1.28), after adjustment for smoking and education. A stronger inverse association was observed in analyses restricted to non-small cell lung cancer (adjusted odds ratio 0.39, 95% confidence interval 0.16-0.96). These results suggest that regular aspirin use might be inversely associated with risk of lung cancer in women, particularly the non-small cell sub-type. PMID- 12085256 TI - Does place of birth influence endogenous hormone levels in Asian-American women? AB - In 1983-87, we conducted a population-based case-control study of breast cancer in Asian women living in California and Hawaii, in which migration history (a composite of the subject's place of birth, usual residence in Asia (urban/rural), length of time living in the West, and grandparents' place of birth) was associated with a six-fold risk gradient that paralleled the historical differences in incidence rates between the US and Asian countries. This provided the opportunity to determine whether endogenous hormones vary with migration history in Asian-American women. Plasma obtained from 316 premenopausal and 177 naturally premenopausal study controls was measured for levels of estrone (E1), estradiol (E2), estrone sulphate (E1S), androstenedione (A), testosterone (T), dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulphate (DHEAS), progesterone (PROG) and sex hormone-binding globulin (SHBG). Levels of the oestrogens and sex hormone-binding globulin did not differ significantly between Asian- and Western-born women, although among premenopausal women, those least westernised had the lowest levels of E1, E2, and E1S. Androgen levels, particularly DHEA, were lower in women born in the West. Among premenopausal women, age-adjusted geometric mean levels of DHEA were 16.5 and 13.8 nmol l(-1) in Asian- and Western-born women respectively; in postmenopausal women these values were 11.8 and 9.2 nmol l(-1), (P<0.001) respectively. Among postmenopausal women, androgens tended to be highest among the least westernised women and declined as the degree of westernisation increased. Our findings suggest that aspects of hormone metabolism play a role in population differences in breast cancer incidence. PMID- 12085257 TI - Human papillomavirus infection as a risk factor for anal and perianal skin cancer in a prospective study. AB - Human papillomavirus has emerged as the leading infectious cause of cervical and other anogenital cancers. We have studied the relation between human papillomavirus infection and the subsequent risk of anal and perianal skin cancer. A case-cohort study within two large Nordic serum banks to which about 760 000 individuals had donated serum samples was performed. Subjects who developed anal and perianal skin cancer during follow up (median time of 10 years) were identified by registry linkage with the nationwide cancer registries in Finland and Norway. Twenty-eight cases and 1500 controls were analysed for the presence of IgG antibodies to HPV 16, 18, 33 or 73, and odds ratios of developing anal and perianal skin cancer were calculated. There was an increased risk of developing anal and perianal skin cancer among subjects seropositive for HPV 16 (OR=3.0; 95%CI=1.1-8.2) and HPV 18 (OR=4.4; 95%CI=1.1-17). The highest risks were seen for HPV 16 seropositive patients above the age of 45 years at serum sampling and for patients with a lag time of less than 10 years. This study provides prospective epidemiological evidence of an association between infection with HPV 16 and 18 and anal and perianal skin cancer. PMID- 12085258 TI - Quantification of the effect of mammographic screening on fatal breast cancers: The Florence Programme 1990-96. AB - Breast cancer cases diagnosed in women aged 50-69 since 1990 to 1996 in the City of Florence were partitioned into those who had been invited to screening prior to diagnosis and those who had not. All cases were followed up for vital status until 31 December 1999. The cumulative number of breast cancer deaths among the cases were divided by screening and invitation status, to give the rates of cancers proving fatal within a period of 8 years of observation (incidence-based mortality). We used the incidence-based mortality rates for two periods (1985-86, 1990-96), pre and during screening. The incidence-based mortality ratio comparing 1990-96 and 1985-86 was 0.50 (95% CI : 0.38-0.66), a significant 50% reduction. For noninvited women, compared to 1985-86, there was a 41% significant mortality reduction (RR=0.59, 95% CI : 0.42-0.82). The comparable reduction in those invited was a significant 55% (RR=0.45, 95% CI : 0.32-0.61). The incidence ratio of rates of cancers stage II or worse was close to one when the noninvited in 1990-96 were compared with 1985-86 (RR=0.97, 95% CI : 0.78-1.21). Excluding prevalent cases, the rate of stage II+ breast cancer cases was 42% lower in Screened women compared with the noninvited (RR=0.58, 95% CI : 0.45-0.74). This study confirmed that new treatments and the first rounds of the screening programme contributed to reducing mortality from breast cancer. PMID- 12085259 TI - Prognostic significance of immunohistochemically detected breast cancer node metastases in 218 patients. AB - Axillary lymph node metastases detected by immunohistochemistry in standard node negative patients with breast carcinomas (13 out of 129 infiltrating ductal carcinomas and 37 out of 89 infiltrating lobular carcinomas) do not have any prognostic significance in patients followed up for a long time (respectively 24 and 18 years). Moreover, their pejorative significance in the literature is debatable since the groups and events taken into account are heterogeneous. PMID- 12085260 TI - High-risk human papillomavirus clearance in pregnant women: trends for lower clearance during pregnancy with a catch-up postpartum. AB - We followed 353 women referred with abnormal cervical cytology in a non intervention cohort study. In 91 pregnant women we compared high-risk human papilloma virus rates in the subsequent trimesters and postpartum in comparison to 262 non-pregnant women. High-risk human papilloma virus clearance was compared with 179 high-risk human papilloma virus positive non-pregnant women. Our main questions were: (1) do high-risk human papilloma virus rates change during pregnancy?; and (2) is there any difference between high-risk human papilloma virus clearance in pregnant and non-pregnant women? Women were monitored 3-4 monthly by cytology, colposcopy, and high-risk human papilloma virus testing. The median follow-up time was 33 months (range 3-74). Non-pregnant women showed prevalence rates of high-risk human papilloma virus of 64, 57, 53, and 50%, respectively, in four subsequent 3-months periods since the start of the study. These high-risk human papilloma virus rates were higher than in the three trimesters of pregnancy, and during the first 3 months postpartum, i.e. 50, 44, 45, and 31%, respectively. Postpartum only, this difference was statistically significant (P=0.004). Paired comparisons of high-risk human papilloma virus prevalence rates of the different trimesters with the postpartum rate showed (McNemar test) decreased rates: first trimester: 18% (P=0.02), second trimester: 13% (P=0.02) and third trimester: 23% (P<0.005). Such a phenomenon was not found in non-pregnant women. Pregnant women showed a trend for increased high-risk human papilloma virus clearance during the third trimester and postpartum compared to non-pregnant women (hazard ratios 3.3 (0.8-13.7) and 4.6 (1.6-12.8), respectively). These results suggest a lowered immune-response against human papilloma virus during the first two trimesters of pregnancy with a catch-up postpartum. PMID- 12085261 TI - Paradoxical expression of cell cycle inhibitor p27 in endometrioid adenocarcinoma of the uterine corpus - correlation with proliferation and clinicopathological parameters. AB - p27 is regarded as a cyclin-dependent kinase inhibitor of the G1-to-S cell cycle progression by suppressing the kinase activity of cyclin/cyclin-dependent kinase complex. This study aimed to investigate p27 expression in the normal endometrium and endometrioid adenocarcinoma of the uterine corpus and the correlation of its expression with cell proliferation and clinicopathological parameters. Tissue samples of 127 endometrioid adenocarcinomas and 15 normal endometria were used in the study. Immunohistochemical staining for detecting p27 and Ki-67 was performed by the labelled streptavidin-biotin method on formalin-fixed and paraffin embedded tissue samples. The expression was given as the labelling index, which indicates the percentage of positive nuclei. p27 staining was observed in the nuclei of the glandular cells in the functional layer of the secretory phase endometrium, whereas it was negative in those of the proliferative phase. In endometrioid adenocarcinomas, the labelling index of p27 expression paradoxically increased more significantly in the higher histological grades and was correlated with that of Ki-67. The high level of p27 expression was associated with clinicopathological parameters such as FIGO stage, lymph node metastasis, lymphovascular space involvement and myometrial invasion. High p27 expression was linked to higher grades of endometrioid adenocarcinoma, cell proliferation and some clinical prognostic factors. These results indicate that p27 might be an indicator of poor prognosis. PMID- 12085262 TI - Localisation of somatostatin and somatostatin receptors in benign and malignant ovarian tumours. AB - Somatostatin has been identified as having anti-proliferative, anti-angiogenic and pro-apoptotic actions in many tumour systems, and these effects are mediated through a family of five transmembrane G-protein coupled SRIF receptors. Ovarian cancer is the commonest gynaecological malignancy in the UK and maintenance therapy is urgently required. Native somatostatin expression and its receptors sst(1,2,3 and 5) were studied with immunohistochemistry in 63 malignant and 35 benign ovarian tumours of various histological types. Fifty-seven out of 63 (90%) of malignant and 26/35 (74%) benign tumours expressed somatostatin. Receptors sst(1,2,3 and 5) were expressed variably in epithelial, vascular and stromal compartments for both benign and malignant tumours. Somatostatin was found to correlate significantly with stromal sst(1) (P=0.008), epithelial sst(1) (P<0.001), stromal sst(2) (P=0.019), vascular sst(2) (P=0.026), epithelial sst(3) (P=0.026), stromal sst(5) (P=0.013) and vascular sst(5) (P=0.038). Increased expression of native somatostatin correlating with somatostatin receptors in malignant ovarian tumours raises the possibility that either synthetic somatostatin antagonists or receptor agonists may have therapeutic potential. PMID- 12085263 TI - Increased expression of survivin in gastric cancer patients and in first degree relatives. AB - Survivin was recently described as an apoptosis inhibitor. Its pathogenic role in gastric cancer is largely unknown. Expression of survivin in gastric cancer and non-cancer first-degree relatives, and its association with apoptosis and cyclo oxygenase-2 expression was investigated. Fifty gastric cancer, 30 non-cancer first-degree relatives, 20 normal controls and five gastric cancer cell lines were studied. Survivin and cyclo-oxygenase-2 were evaluated by reverse transcriptase-polymerase chain reaction, immunohistochemistry and Western blot. Survivin expression was absent from normal gastric mucosa. All five cancer cell lines and 34 out of 50 (68%) human gastric cancer tissues expressed survivin mRNA. Survivin expression was less frequent (22%; P<0.001) in adjacent non-tumour gastric tissues. Immunohistochemistry and Western blot obtained similar findings. Gastric cancers with survivin expression displayed significantly reduced apoptosis (P=0.02), and associated with cyclo-oxygenase-2 overexpression at both mRNA (P=0.001) and protein levels (P=0.041). Moreover, survivin mRNA was detected in the gastric mucosa of eight (27%) non-cancer relatives. Expression in non cancer patients showed positive correlation with H. pylori infection (P=0.004). This demonstrates the frequent expression of survivin in gastric cancer and in first-degree relatives. Co-expression of survivin and cyclo-oxygenase-2 may suggest multiple pathways contributing to the inhibition of apoptosis in gastric cancer. PMID- 12085264 TI - Novel substituted methylenedioxy lignan suppresses proliferation of cancer cells by inhibiting telomerase and activation of c-myc and caspases leading to apoptosis. AB - Conventional solvent fractionation and bioactivity based target assays were used to identify a new anti-cancer molecule from Phyllanthus urinaria, a herbal medicinal plant used in South India. At each step of the purification process the different fractions that were isolated were tested for specific anti proliferative activity by assays measuring the inhibition of [(3)H]thymidine incorporation, and trypan blue drug exclusion. The ethyl acetate fraction that contained the bioactivity was further purified and resolved by HPLC on a preparative column. The purity of each of the fractions and their bioactivity were checked. Fraction 3 demonstrated a single spot on TLC and showed maximum anti-proliferative activity. This fraction was further purified and the structure was defined as 7'-hydroxy-3',4',5,9,9'-pentamethoxy-3,4-methylene dioxy lignan using NMR and mass spectrometry analysis. The pure compound and the crude ethyl acetate fraction which showed anti-proliferative activities were examined for ability to target specific markers of apoptosis like bcl2, c-myc and caspases and for effects on telomerase. Four specific cancer cell lines HEp2, EL-1 monocytes, HeLa and MCP7 were used in this study. The results indicate that 7'-hydroxy 3',4',5,9,9'-pentamethoxy-3,4-methylene dioxy lignan was capable of inhibiting telomerase activity and also could inhibit bcl2 and activate caspase 3 and caspase 8 whose significance in the induction of apoptosis is well known. We believe that this compound could serve as a valuable chemotherapeutic drug after further evaluations. PMID- 12085265 TI - Cytotoxic targeting of F9 teratocarcinoma tumours with anti-ED-B fibronectin scFv antibody modified liposomes. AB - We prepared small unilamellar liposomes derivatised with single chain antibody fragments specific for the ED-B domain of B-fibronectin. This extracellular matrix associated protein is expressed around newly forming blood vessels in the vicinity of many types of tumours. The single chain antibody fragments were functionalised by introduction of C-terminal cysteines and linked to liposomes via maleimide groups located at the terminal ends of poly(ethylene glycol) modified phospholipids. The properties of these anti-ED-B single chain antibody fragments-liposomes were analysed in vitro on ED-B fibronectin expressing Caco-2 cells and in vivo by studying their biodistribution and their therapeutic potential in mice bearing subcutanous F9 teratocarcinoma tumours. Radioactively labelled ((114m)Indium) single chain antibody fragments-liposomes accumulated in the tumours at 2-3-fold higher concentrations during the first 2 h after i.v. injection compared to unmodified liposomes. After 6-24 h both liposome types were found in similar amounts (8-10% injected dose g(-1)) in the tumours. Animals treated i.v. with single chain antibody fragments-liposomes containing the new cytotoxic agent 2'-deoxy-5-fluorouridylyl-N(4)-octadecyl-1-beta-D arabinofuranosylcytosine (30 mg kg(-1) per dose, five times every 24 h) showed a reduction of tumour growth by 62-90% determined on days 5 and 8, respectively, compared to animals receiving control liposomes. Histological analysis revealed a marked reduction of F9 tumour cells and excessive deposition of fibronectin in the extracellular matrix after treatment with single chain antibody fragments-2 dioxy-5-fluorouridylyl-N(4)-octadecyl-1-beta-D-arabinofuranosylcytosine liposomes. Single chain antibody fragments-liposomes targeted to ED-B fibronectin positive tumours therefore represent a promising and versatile novel drug delivery system for the treatment of tumours. PMID- 12085266 TI - Transforming Growth Factor-beta2 protects the small intestine during methotrexate treatment in rats possibly by reducing stem cell cycling. AB - During chemo- and radiation therapy, the balance between epithelial cell proliferation, differentiation, and cell death at the villus tip is disrupted by premature death of dividing epithelial cells. This will subsequently lead to the onset of mucosal barrier injury in the whole gastrointestinal tract. Up till now there is no validated method to treat side effects occurring due to therapy. An approach to manage this side effect might be to reversibly arrest growth of epithelial stem cells during therapy using Transforming Growth Factor-beta2. A Transforming Growth Factor-beta2 enriched fraction prepared from bovine milk was shown to protect small intestinal epithelial cells against cell cycle specific chemotherapeutic agents by arresting the cells in G1-phase. Secondly, in a rat model for induced small intestinal damage, oral supplementation of rats exposed to methotrexate with the Transforming Growth Factor-beta2 enriched fraction significantly reduced the chemotherapy-associated weight loss and ileal villus atrophy by reducing cell proliferation in the normal stem cell population. Thus oral supplementation with a bovine milk fraction enriched for Transforming Growth Factor-beta2 attenuated the side effects of chemotherapy in the small intestine in rats. PMID- 12085269 TI - [Local treatment of liver metastases--multidisciplinary polypragmasia or interdisciplinary strategy?]. PMID- 12085270 TI - [European vascular surgery--do we need a German congress?]. PMID- 12085267 TI - Inhibition of renal cell carcinoma angiogenesis and growth by antisense oligonucleotides targeting vascular endothelial growth factor. AB - Angiogenesis is critical for growth and metastatic spread of solid tumours. It is tightly controlled by specific regulatory factors. Vascular endothelial growth factor has been implicated as the key factor in tumour angiogenesis. In the present studies we evaluated the effects of blocking vascular endothelial growth factor production by antisense phosphorothioate oligodeoxynucleotides on the growth and angiogenic activity of a pre-clinical model of renal cell carcinoma (Caki-1). In vitro studies showed that treating Caki-1 cells with antisense phosphorothioate oligodeoxynucleotides directed against vascular endothelial growth factor mRNA led to a reduction in expressed vascular endothelial growth factor levels sufficient to impair the proliferation and migration of co-cultured endothelial cells. The observed effects were antisense sequence specific, dose dependent, and could be achieved at a low, non-toxic concentration of phosphorothioate oligodeoxynucleotides. When vascular endothelial growth factor antisense treated Caki-1 cells were injected into nude mice and evaluated for their angiogenic potential, the number of vessels initiated were approximately half that induced by untreated Caki-1 cells. To test the anti-tumour efficacy of vascular endothelial growth factor antisense, phosphorothioate oligodeoxynucleotides were administrated to nude mice bearing macroscopic Caki-1 xenografts. The results showed that the systemic administration of two doses of vascular endothelial growth factor antisense phosphorothioate oligodeoxynucleotides given 1 and 4 days after the tumours reached a size of approximately 200 mm(3) significantly increased the time for tumours to grow to 1000 mm(3). PMID- 12085271 TI - [Changing surgical therapy because of clinical studies?]. AB - The randomised controlled clinical trial (RCT) is a powerful instrument to evaluate different therapeutic regimens. In a survey among 115 physicians visiting the 25th annual meeting of the Surgical Society of Berlin and Brandenburg, the RCT was judged to be very important when changes of therapeutic strategies are discussed. 90 % of all participants claimed to use data from RCTs in the clinical routine and 89 % would participate in such a trial. In official (e. g. discussions during coffee breaks at scientific meetings) or non-medical (e. g. non-scientific press or media) sources of information were assessed as irrelevant for decisions regarding therapeutic strategies. However, in contrast to this view laparoscopic cholecystectomy was introduced into clinical practice rapidly because patients informed by external (non-medical) sources preferred to be operated on with the "modern" technique. Clinical trials with a high level of evidence had no relevant influence on the rapid distribution of laparoscopic cholecystectomy. Controversial discussions concerning the extent of lymphadenectomy with gastric resection for carcinoma demonstrate that the value of excellent clinical RCTs is low if their results challenge a stable paradigma of the surgical scientific society. To allow a rational judgement, new surgical technologies should undergo a scientific gradual evaluation in agreement with the principles of evidence based medicine. PMID- 12085272 TI - [The role of preoperative oesophageal manometry in patients with gastro oesophageal reflux disease]. AB - BACKGROUND: Oesophageal manometry has become a standard investigation in the management of functional disorders of the oesophagus. However, the importance of the various manometry parameters for the surgical management of patients with gastro-oesophageal reflux disease (GERD) has not yet been studied adequately. AIM: This study examined the importance of manometric findings on surgical treatment results in patients with GERD. PATIENTS AND METHODS: Between January 1995 and January 2000, 123 consecutively referred patients with GERD symptoms were studied. Ninety-seven of these (52 women and 45 men, mean age 48 years) underwent Nissen fundoplication in conventional or laparoscopic technique. Preoperatively, each patient received stationary oesophageal manometry with a water perfusion catheter, gastroduodenoscopy and 24h-pH-metry of the oesophagus. Independent of manometric results, patients received 360 degrees fundoplicatio with 2 cm cuff ("floppy Nissen"). RESULTS: Eighty-five of the 97 patients (88 %) had an incompetent lower oesophageal sphincter. Hypomotility was diagnosed in 17 (18 %). Amotile achalasia and diffuse oesophageal spasms were seen in two and one patient, respectively, who were not operated on. Follow-up data were available for 92 % of patients (mean follow-up time 1.8 years). Five and 8 patients (1 and 2 of these with hypomotility) postoperatively developed persistent dysphagia and GERD recurrence, respectively (relative risks 1.2 and 1.6). Two patients (one with hypomotility) required surgical revision. CONCLUSIONS: Preoperative oesophageal manometry in GERD patients is useful only for determining the position of the upper boundary of the lower oesophageal sphincter and for ruling out specific motility disorders. In patients with oesophageal hypomotility, "floppy Nissen" fundoplicatio can be performed without an excess risk of dysphagia. PMID- 12085274 TI - [Comparison between treatment results for gastric cancer in younger and elderly patients]. AB - INTRODUCTION: While gastric cancer shows an increased incidence in elderly patients, the rate of younger patients affected by this disease represents up to 15 %. Younger patients are frequently diagnosed with advanced tumor stages with a poor prognosis although literature data on this issue are controversial. PATIENTS AND METHODS: 643 patients with primary gastric carcinoma were operated in our institution between March 1986 and December 2000. No neoadjuvant treatment was administered in these patients. We analysed the data of these patients retrospectively. A comparison of the results between patients younger than 40 years (n = 38, median age 37 years) and older than 70 years (n = 182, median age 75 years) was performed. RESULTS: The radical (R0-) resectability rate was rather high for both, younger (78.9 %) and elderly (76.9 %) patients. Postoperative morbidity was higher in elderly than in younger patients (32.9 % vs. 23.2 %; p < 0.05), as well as the postoperative mortality (7.7 % vs. 2.6 %; p < 0.05). Both younger and elderly patients showed advanced (II to IV) tumor stages (76.3 % vs. 73.3 %, n. s.). There was a significant difference between the rate of diffuse carcinomas in young and elderly patients (63.2 % vs. 22.5 %). The 5-years survival rate following R0-resection was significantly higher for younger patients (54.2 % vs. 32.9 %; p = 0.01), differences occurred only after the second postoperative year. CONCLUSIONS: The resectability of gastric carcinoma is not related to the patients age. Due to comorbidity, postoperative morbidity may be increased in elderly patients. Although both younger and elderly patients show advanced tumor stages, diffuse carcinomas are more frequent in younger patients. The short-term prognosis is similar for both age groups, long-term results are better for younger patients. The different life expectancy should be considered when interpreting these results. PMID- 12085275 TI - [Cryotherapy for primary and secondary liver tumours]. AB - AIMS: We assessed the morbidity, mortality and efficacy of cryotherapy for primary and secondary malignant liver tumours in a prospective case control study. METHODS: Since 1996 we performed 77 cryosurgical procedures on 71 patients. 6 patients had hepatocellular carcinoma, the remainders metastases, mainly of colorectal origin (n = 49). Cryotherapy was used when a complete tumour resection was not feasible, but complete tumour destruction by cryotherapy seemed possible. Mean follow-up was 21 months. RESULTS: The mean number of lesions per patient was 2.6 (1-10) with a mean maximum diameter of 3.7 cm (1-11). In 36 of 71 patients cryotherapy was used in combination with liver resection. Morbidity and hospital mortality were 23 % and 2.8 %. In 25 of 34 patients with colorectal primary and preoperatively elevated CEA it returned to normal postoperatively (74 %). 40 of the 60 patients with "R0-treatment" developed tumour recurrence, 10 of these at the cryosite (17 %). Median survival for all patients (colorectal metastases) was 28 (29) months with a 3- and 5-year survival rate of 38 % (43 %) and 30 % (33 %), respectively. CONCLUSION: Cryotherapy is an effective parenchyma sparing method for the treatment of malignant liver tumours in a curative approach. It can be performed safely and a long-term local tumour control is achieved in a high percentage of patients. Survival rates following cryotherapy appear comparable to the results of liver resection. It seems justified to conduct a prospective randomised trial comparing liver resection and cryotherapy for resectable tumours. PMID- 12085276 TI - [Management of common bile duct stones--Results of a nationwide survey with analysis of 8 433 common bile duct explorations in Germany]. AB - BACKGROUND: Aim of this study was the evaluation of the management of choledocholithiasis and outcome of laparoscopic as well as open cholecystectomy in Germany. METHODS: A written questionnaire was sent to 449 randomly selected German surgeons annually from 1991 to 1994 and additionally to all German university hospitals until 1998. RESULTS: A total of 98 482 operations for gallstone disease including 86 485 cholecystectomies (non-university hospitals 1991-1993: n = 60 246, university hospitals 1991-1996: n = 26 239) and 8 433 common bile duct (CBD) explorations (non-university hospitals: n = 6 919, university hospitals: n = 1 514) with or without cholecystectomy were reported. The overall complication rate for CBD explorations was 13.2 % (non-university hospitals) and 15.1 % (university hospitals), the overall hospital mortality rate was 0.64 % (non-university hospitals) and 0.58 % (university hospitals, n. s.). When choledocholithiasis was suspected, in 1998 all university hospitals used a two-stage management with preoperative ERC. In case of diagnosed isolated choledocholithiasis 93 % usually chose an endoscopic approach. When simultaneous cholecysto-choledocholithiasis was diagnosed preoperatively 86 % of the university hospitals used a two-stage management with preoperative ERC and stone extraction and secondary cholecystectomy (1991: 45 %). The percentage of CBD explorations decreased continuously from 7.4 % in 1991 to 3.8 % in 1996 (p < 0.01). CONCLUSION: These results allow for the estimation of frequency and overall risks in surgical therapy of CBD stones. At the moment, laparoscopic CBD exploration does not play a significant role in Germany. Data show a trend to the two-stage "therapeutical splitting" with lower complication rates. PMID- 12085278 TI - [Appendicitis in the last decade of the 20th century--Analysis of two prospective multicenter clinical observational studies]. AB - AIMS OF THE STUDY: The authors present an accurate and comprehensive snapshot of appendicitis in the last decade of the 20 (th) century. The present study also aims to determine the influence of introduction of laparoscopic techniques on the quality of appendicitis therapy. PATIENTS AND METHODS: All consecutive patients admitted for suspected appendicitis were recruited into this prospective multicenter study. Two different periods of time were considered: from 1. 7. 1988 to 31. 12. 1989, 6 266 patients from 18 hospitals were included, and from 1. 6. 1996 to 31. 5. 1997, 7 398 patients from 34 institutions. RESULTS: In the first period (1988/89), 1 869 patients were treated conservatively (29.8 %), 4 397 (70.2 %) were operated. Histopathological analysis of the appendix documented no acute inflammation in 28.8 %, the perforation rate was 8.3 % and the wound infection rate 9.6 %. In the second period (1996/97), 2 430 patients were treated conservatively (32.8 %), in 24.9 % of all operated cases, the histopathological examination shows no acute inflammation of the appendix, the perforation rate was 5.9 % and the rate of wound infections 2.5 %. During this second period, 33.2 % of all appendectomies were performed using the laparoscope, the conversion rate was 7.2 %. CONCLUSIONS: The introduction of laparoscopic techniques did not modify the operative indications for treatment of appendicitis. An increase in the number of normal appendices removed was not observed. The number of perforations and the incidence of wound infections was significantly reduced in the second period. PMID- 12085280 TI - [Results of the surgical therapy in advanced colorectal cancer]. AB - INTRODUCTION: In every 7th patient with colorectal cancer tumor has already spread beyond intestinal wall into surrounding organs. PATIENTS AND METHODS: Between 01. 01. 1990 and 31. 12. 1998 763 patients with colorectal cancer were treated at our surgical department. 166 patients (23 %) presented with tumor contiguous or adherent to adjacent organs (cT4). RESULTS: In most cases tumor was localized in colon (109 patients, 66 %), in 57 patients (34 %) tumor was found in rectum. Potenzial curative resection (R0) was possible in 67 patients (40 %). 66 patients (40 %) had microscopic (R1) or gro beta residual disease (R2) and in 33 patients only palliative surgery was possible. Extended resection of adjacent organs was performed in 97 % in the group with curative resection. 11 patients (8 %) died after multivisceral resection. The 5-year survival for curative resection was 57 %, for patients with microscopic or gro beta residual disease 9 months and for palliative surgery only 4 months. CONCLUSION: Optimistic longterm results in advanced colorectal cancer can only be achieved after curative resection. After incomplete resection or palliative surgery median life expectancy is extremely poor. PMID- 12085279 TI - [Which imaging in case of sigmoid diverticulitis? The value of ultrasound (Conventional B-mode in combination with hydrocolonsonography and colour flow Doppler) in comparison to the well-established modalities like contrast enema and helical computertomography]. AB - Sigmoid diverticulitis is in case of complications like perforation, abscess and peritonitis a life-threatening disease. A diagnostic work up with high sensitivity is mandatory. In a prospective study upon 247 patients with the possible diagnosis of acute diverticulitis ultrasound is a screening method with a high specificity (97 %) and leads in combination with the hydrocolonsonography and the colour flow doppler to a high sensitivity (76 %). Without any typical findings in ultrasound, helical CT scan is the best method in the evaluation of the acute diverticulitis (sensitivity and specificity 100 %), because contrast enema is inferior to CT in the evaluation of abscesses (6 %) and perforation (53 %). CT has in case of perforation and of abscesses a sensitivity of 90 % and 100 %, while ultrasound has a sensitivity of 12 % to perforation and of 39 % to abscesses. PMID- 12085281 TI - [Laparoscopic treatment of iatrogenic perforations of the colon]. AB - Iatrogenic perforations of the colon are serious complications that have been observed in association with both diagnostic and therapeutic colonoscopies, and are not always avoidable, even in the hands of an experienced investigator. Patients suffering from a perforation during elective colonoscopy are starving, and have a clean bowel. If the perforation is quickly diagnosed with only local mild peritonitis, laparoscopic treatment is often possible. The therapeutic options available are: oversewing, wedge resection and, in the individual case, tubular resection, as well as laparoscopic-assisted reconstruction, if indicated including a protective stoma. The decision to perform a laparoscopic procedure should depend on the time of detection of the lesion, and on the skills of the existing surgical team; otherwise, a laparotomy is indicated. We describe four cases that reflect the relevance of the laparoscopic approach. PMID- 12085282 TI - [Therapy of the pilonidal sinus--Primary wound closure or open wound after excision]. AB - The postoperative follow up of 177 operations in 162 patients with pilonidal sinus was investigated. After excision of the sinus a primary wound closure was performed in 80 cases; in 83 cases the wound was left open. After primary wound closure 40 % of the patients showed a primary healing of the wound. Although the remaining 60 % of the wounds healed secondarily the patients were not disabled longer and did not demonstrate more frequently recurrences than those with open wound management. As a consequence we recommend a primary wound closure after excision of a pilonidal sinus. PMID- 12085284 TI - [Significance of pelvic lymphadenectomy for the prognosis after radical cystectomy]. AB - OBJECTIVE: In contrast to other carcinomas such as breast or colon cancer, there are no guidelines regarding the number and location of lymph nodes to be removed during radical surgery in patients with invasive bladder carcinoma. The therapeutic effect of pelvic lymphadenectomy and its influence on tumour staging has not been documented yet. METHODS: Here we present an evaluation of pelvic lymph nodes from 484 patients who underwent radical cystectomy with curative intention between 1986 and 1999. The number of lymph nodes was correlated with the depth of invasion of the primary tumour, occurrence of nodal metastases, clinical outcome, the operating surgeon, and the pathologist. RESULTS: There were 484 patients with a mean age of 62.7 years. Clinical follow up was available from 321 patients with a mean follow up period of 35.9 months. The average number of lymph nodes removed was 14.3 (range: 1-46). The number of lymph nodes removed varied significantly between different surgeons and did not correlate with the pathologists. There was a significant correlation between the number of lymph nodes removed and the tumour-free survival in pT2 or pT3 tumours and in patients without lymph node metastases. Multivariate analysis revealed that pT-category (p < 0.01), pN-category (p < 0.01), and the total number of lymph nodes removed (p = 0.04) were the most important factors affecting survival. CONCLUSION: The more extensive lymphadenectomy significantly improved the prognosis of patients with invasive bladder cancer and therefore, represents a potentially curative procedure. The results indicate a need for a standardised lymph node dissection. PMID- 12085285 TI - [Retrogastral located gastrointestinal stromal tumor (GIST) as a sonographically detected rare incidental finding]. AB - Gastrointestinal stromal tumors (GIST) represent compared to carcinomas a rare group of neoplasias of the gastro-intestinal tract of unclear dignity. We report the example of a patient suffering from a big retrogastral located gastrointestinal stromal tumor which had been detected as an incidental finding without previous complaints. Because origin and dignity of the process could not definitely be diagnosed, total resection (R0-resection) without systemic lymphadenectomy of the process measuring 11.5 cm x 11 cm x 7 cm was performed. Mitotic activity and tumor-size are regarded as predictive factors of potenzial malignancy of GISTs. In general tumors with low mitotic activity of up to 5 mitoses per 50 high power fields (HPFs) and a diameter smaller than 5 cm are regarded as benign. In the presented case, up to 4 mitoses per 50 HPFs could be detected and thus, in connection with tumor-size, an uncertain biological behaviour of the process has to be expected. Since no generally accepted consensus on the treatment of the GISTs exists, also patients originally suffering from tumors regarded as borderline-malignant should undergo a close meshed follow-up in regular intervals. PMID- 12085286 TI - [Spontaneous rupture of hepatic focal nodular hyperplasia--a rare cause of acute intraabdominal bleedingf]. AB - Benign hepatic mass lesions are rare. Haemangioma, focal nodular hyperplasia and hepatocellular adenoma represent the most common ones. For the reason of serious clinical complications and potential malignant transformation of hepatic adenoma, surgical resection is the therapy of choice. Unlike adenoma focal nodular hyperplasia is mostly asymptomatic and rarely causes severe complications. We report on a spontaneous rupture and life-threatening intraperitoneal haemorrhage of a large, asymptomatic FNH, where surgical resection was indicated. Within three month after hepatic resection and after termination of oral contraceptives almost complete regeneration of liver tissue and dramatic regression of a second FNH lesion was apparent. Although haemorrhage of hepatic FNH is extremely rare, this case highlights the small risk of rupture in large lesions. PMID- 12085287 TI - [Acute hemorrhage from Meckel's diverticulum--Laparotomy or laparoscopy?]. AB - Meckel's diverticulum is the most common congenital malformation of the gastrointestinal tract with a potential risk to develop complications such as obstruction, diverticulitis or intussusception. Lower gastrointestinal bleeding due to ulceration of heterotopic gastric tissue of the diverticulum is a known phenomenon in children and young adults. We present two cases of a 15-year-old girl and a 20-year-old man that revealed a massive lower gastrointestinal hemorrhage of unknown origin. In this emergency situation laparotomy was performed in combination with lower endoscopy as rendezvous manouver. In both cases a Meckel's diverticulum with peptic ulceration was the source of hemorrhage, in one case the bleeding was active and visible. After resection of a short small bowel segment and end-to-end anastomosis the postoperative course was uneventful. We prefer in the case of lower gastrointestinal hemorrhage with hemodynamic instability laparotomy with intraoperative endoscopy instead of laparoscopy. PMID- 12085288 TI - [Prospective multi-center study colon/rectum carcinoma (primary tumors)--Results of the year 2000]. AB - INTRODUCTION: Surgical quality control in the form of multicentre studies make it possible to analyse the treatment status of a given surgical illness under quality assurance aspects. MATERIAL/METHODS: On 1. 1. 2000, under the patronage of the Convent of Hospital Chief Surgeons a project (ongoing) - organised and conducted by the Institute for Quality Management in Operative Medicine at the Otto-von-Guericke University of Magdeburg - was initiated to document and collect the data of patients with colorectal cancer throughout the whole of Germany. This work is supported by the German Society of Surgery, and the Surgical Working Group, Quality Assurance within the German Society of Surgery. Currently, some 282 hospitals throughout the country are involved in establishing a prospective documentation of colorectal carcinoma. Participation in this study is on a voluntary basis. The anonymity of both patients and hospitals is guaranteed, and no hospitals wishing to participate are excluded. Both operatively and conservatively treated patients are being documented, and no randomization takes place. RESULTS: In the year 2000, the participating hospitals documented a total of 9 477 patients with a colorectal carcinoma, including 6 975 patients with a carcinoma of the colon, and 3 402 with a rectal carcinoma. The average age of the patients was 68.5 years, and there were 5 010 men and 4 467 women. The operation rate was 99.2 %, the resection rate 95.6 %. The abdominoperineal resection rate was 27.4 %. The indicators for diagnostic quality as set out by Hermanek were largely complied with, but some deviations were noted. DISCUSSION: On the basis of the data collected, structures were established for a uniform Germany-wide quality management for a clinical condition with a major health policy impact. This information make it possible for the hospitals to identify and eliminate deficits in the structural and process quality in the diagnosis and treatment of colorectal cancer, and in this way to improve outcome quality. This means that the results of medical care research have an immediate impact on the individual treatment received by a given patient. PMID- 12085290 TI - [Extracorporeal shock wave therapy in orthopedics--costs and benefits]. PMID- 12085291 TI - [Musculoskeletal shock wave therapy--current database of clinical research]. AB - During the past decade application of extracorporal shock waves became an established procedure for the treatment of various musculoskeletal diseases in Germany. Up to now the positive results of prospective randomised controlled trials have been published for the treatment of plantar fasciitis, lateral elbow epicondylitis (tennis elbow), and of calcifying tendinitis of the rotator cuff. Most recently, contradicting results of prospective randomised placebo-controlled trials with adequate sample size calculation have been reported. The goal of this review is to present information about the current clinical database on extracorporeal shock wave treatment (ESWT). PMID- 12085292 TI - [Extracorporal shock wave treatment of the achilles tendinitis: Experimental and preliminary clinical results]. AB - OBJECTIVE: Various studies have shown the benefit of extracorporal shock wave therapy (ESWT) in the treatment of soft tissue pathologies. The aim of this prospective study was to compare the results of ESWT with the operative treatment for achilleus tendopathy. Furthermore mechanical effects of the shock waves on the achilleus tendon were analysed by an in-vitro model. METHOD: The achilleus tendons of common pigs treated with 1500 impulses twice using an energy varying from 0.23 - 0.54 mJ/mm (2) were histologically examined. The clinical results of two patient groups of which the first (n = 28) was treated with 2000 impulses (0.23 mJ/mm (2)) once and the second (n = 26) underwent surgery for achilleus tendopathy were compared. RESULTS: The in-vitro model demonstrated that an EFD of 0.42 and 0.54 mJ/mm (2) can lead to tendon lesions. One year follow-up showed good and excellent results in 69 % and satisfactory results in 15 % of the operated group and good and excellent results in 29 % and satisfactory results in 43 % of the ESWT group. We could show a time-dependent effect of the shock wave treatment on the clinical outcome. CONCLUSIONS: Follow-ups show acceptable results with little side effects when an energy flow density (EFD) under 0.23 mJ/mm (2) is used. ESWT offers a non-invasive therapeutic concept that can seriously be contemplated before operative treatment, but lower success rates in comparison to the operative results can be expected. PMID- 12085293 TI - [Extracorporeal shock waves induce ventral-periosteal new bone formation out of the focus zone--results of an in-vivo animal trial]. AB - AIM: It is known that high-energy extracorporeal shock waves, focussed to the intact distal rabbit femur, induce periosteal new bone formation within the focus zone. This study was done to investigate whether there is additionally new bone formation outside of the focus zone, i. e., in the proximal femur, 10 days after shock wave application to the distal rabbit femur. METHOD: 18 adult Chinchilla Bastard rabbits were randomly divided into three groups (A, B, C; each n = 6). One distal femur of each animal was focussed for shock wave application with different positive energy flux densities (A 1.2 mJ/mm (2), B 0.9 mJ/mm (2), C sham treatment). Only the proximal femur outside the shock wave focus was investigated. Here periosteal new bone formation was evaluated by fluorescent labelling. The maximum thickness was measured by means of computer-assisted image analysis. RESULTS: Maximum ventral-periosteal new bone formation outside the shock wave focus was significantly increased in group A compared to group B (p = 0.007) or group C (p = 0.001). In contrast, the maximum ventral-periosteal new bone formation outside the shock wave focus demonstrated no statistically significant difference in group B compared to group C (p = 1.0). CONCLUSION: Depending from the amount of positive energy flux density applied to distal rabbit femur a significantly increased ventral-periosteal new bone formation results in the proximal rabbit femur outside the shock wave focus. PMID- 12085294 TI - [The transitory bone marrow edema syndrome of the hip]. AB - AIM OF THE STUDY: Since MRI-studies had begun to establish the diagnosis of transitory bone marrow edema syndrome of the hip orthopedic surgeons have tried to integrate this new syndrome into the internationally accepted system of musculoskeletal diseases. Particularly, the relation to non-traumatic osteonecrosis of the femoral head and the possibilities in therapy were investigated in our clinical trial. METHODS: Our clinical trial encompassed 106 patients suffering from the transitory bone marrow edema syndrome diagnosed in our department between the years 1985 and 2000. In order to confirm this diagnosis we used the patients' histories, their clinical courses, MRI studies, scintigraphic bone scans, intraosseal pressure measurements, phlebographies, laboratory data, and histologic specimens. One half of our collective positive for transient bone marrow edema of the hip underwent core-decompression surgery (50 patients), the other half (56 patients) was treated conservatively by analgesic medication combined with restriction of weight-bearing in the affected extremity. RESULTS: Patients positive for transitory bone marrow edema syndrome of the hip are middle-aged individuals with a male to female predominance of 60 : 40. This group has no or only few risk factors usually associated with osteonecrosis of the femoral head. Thus, the missing alcoholic abuse is striking. All patients suffering from transitory bone marrow edema syndrome of the hip recovered completely independent of the therapy we initiated and none of them showed any signs of osteonecrosis. The one half undergoing surgical decompression of the edema by using a 4.5 mm drill experienced an markedly accelerated relief of their clinical symptoms as well as their signal changes on MRI studies. Conventional X-ray pictures and scintigraphic bone scans are not useful for early differentiation between early stages of osteonecrosis and bone marrow edemas. This also accounts for the historical measurements of intraosseal pressure determinations and phlebographies. In contrast to that, MRI studies are effective in early differentiation between osteonecrosis and bone marrow edema syndrome of the hip, especially when contrast medium (gadolinium) is administrated intravenously and fat-suppressed MRI-sequences find use. Beginning osteonecrosis of the femoral head shows a segmental loss of contrast medium, a "double line sign" interface to the intact bone marrow, and only in a few cases they are associated with a huge symptomatic edema. The histologic examination of specimens obtained from 43 patients with transitory bone marrow edema syndrome of the hip revealed no signs of osteonecrosis. CONCLUSION: MRI studies are useful in differentiation between bone marrow edema syndrome of the hip and non-traumatic osteonecrosis of the femoral head in each stage of these two diseases. The thorough differentiation between these two diseases is of extraordinary importance for the clinical work-up of the patients as well as for scientific reasons. The course of primary bone marrow edema is benign as it results in entire recovery. The core decompression surgery offers the chance to shorten the course of the disease. PMID- 12085295 TI - [Spontaneous bilateral fractures of the femoral neck. Etiology-frequency diagnosis-therapy]. AB - INTRODUCTION: Medial and pertrochanteric fractures of the femoral neck occur commonly in patients with bone weakness due to osteoporosis or osteomalacia. Combined bilateral hip injury is much rarer. PATIENTS AND METHODS: The case of a Turkish woman with bilateral fractures of the femoral neck due to secondary hyperparathyroidism caused by nutritional vitamin-D 3 deficiency is described and compared to 80 cases of bilateral hip fractures mentioned in English and German literature. RESULTS: Until the late 1950's the most common reason for simultaneous bilateral femoral neck fractures was convulsive therapy of psychosis by drug or electroshock without use of relaxants, followed by seizures of other origin. Primary and secondary bone diseases are the most common reason for two fold spontaneous bilateral fractures of the femoral neck. CONCLUSION: At the time of first pain radiological findings are often normal. In this situation MRI, isotope bone scanning or additional chemical diagnostics have proved to be useful. The medical regulation of the bone disease is of highest priority. On account of the pain, bone stability and deformation, conservative and operative therapy such as osteosynthesis or THR has to be performed. PMID- 12085296 TI - [Clinical and radiological results of individual hip stems of the type Adaptiva(R) without cement]. AB - AIM: Long-term anchorage of foreign material in vital bone has proven to be the main problem in hip arthroplasty. Bone cement, a material for filling and fitting, allows an excellent solution for older people. Many failures have been blamed on the use of polymethylmethacrylate in younger patients. In our opinion, modelling a stem to the individual anatomic needs of younger patients and to implant it without cement but with a stable press-fit is a good way to transmit stress harmoniously from the prosthesis to the bone and to obtain a long-lasting function. This individual hip stem is now available in the third generation under the name Adaptiva(R). We would like to present our first results. METHOD: Between October 1993 and September 1995 150 individual hip prosthesis of the Adaptiva(R) type have been implanted. In the average the patients were 53,2 years old. The average time of follow-up was 19.9 (12 to 44) months. RESULTS: The Merle d'Aubigne score showed excellent absolute and relative results for pain, mobility and ability to walk. No aseptic loosening of the stem occurred. CONCLUSION: Our early results are promising, but we have to wait for the long-term results, which are part of a current study. PMID- 12085297 TI - [Computer-based motion simulation of total hip prostheses with ceramic-on-ceramic wear couple. Analysis of implant design andorientation as influence parameters]. AB - OBJECTIVE: In THA, ceramic-on-ceramic wear couples are increasingly used. A restricted range of motion (ROM) due to unfavourable implant design or positioning may cause impingement or dislocation, which can result in failure of ceramic inserts. METHODS: By means of a 3-D CAD program different hip joint movements were simulated and the effects of ceramic hip implant design and position on the range of motion were analysed. RESULTS: To offer sufficient ROM and to minimise risk of impingement and dislocation, inclination angle of the acetabular cup should be 45 degrees, cup anteversion 15 degrees and stem antetorsion 0 degrees to 10 degrees. In regard to implant design, acetabular cups with slightly- recessed ceramic inserts should be used. Prosthetic systems with an elevated liner or with a mushroom-shaped femoral head are associated with limited ROM and increased risk of mechanical failure. The ratio of head to neck diameter should never be less than 2 : 1. Larger heads not only increase ROM, but also the stability of the prosthesis against dislocation. Thereby, the wear rate of ceramic-on-ceramic couples is not increased, in contrast to polyethylene. CONCLUSION: Considering certain criteria for ceramic hip implants regarding implant positioning, design and handling, ceramic-on-ceramic couples can be used with low risk of revision surgery and they can also reduce the prosthesis loosening associated with wear in young and active patients. PMID- 12085298 TI - [Prophylaxis of heterotopic ossification in hip revisions with 7 Gy single-dose radiation]. AB - AIM OF STUDY: Radiation therapy is an established method for the prevention of heterotopic ossification. The aim of this study was to assess whether radiation therapy is also effective in revision arthroplasties. METHOD: 143 hips were irradiated with 7 Gy anterior-posterior applied single dose radiation, on one of the first four days after revision surgery. RESULTS: After an average follow-up of 18 months, 107 hips were evaluated radiographically using Brooker's criteria. In comparison to the immediate postoperative findings (26 % showed no heterotopic ossification, 66 % had Brooker I, 5 % Brooker II, and 3 % Brooker III), at final follow-up 19 % showed no heterotopic ossification, 66 % had Brooker I, and 7.5 % Brooker II and III. New ossification or an increase of one grade was found in 12 %, of two grades in 4 %. Grade 0 showed significantly more ossifications. Gender (p = 0.43), age (p = 0.43), the type of revision surgery (p = 0.36), the day of radiation (1st day vs. 2nd, 3rd, 4th day; p = 0.46) had no significant influence on this increase. There was no correlation between pain score and Brooker grades (p = 0.755). There was a significantly (p < 0.01) negative correlation (r = - 0.31) between the Brooker grades and flexion. CONCLUSION: Radiation therapy with 7 Gy single dose effectively prevents the new formation of heterotopic ossification or the progression of ossifications after revision surgery. To prevent a decrease in the range of motion due to HO after revision surgery, HO should be removed intraoperatively. PMID- 12085299 TI - [Periprosthetic osseous changes 2, 4, and 6 years after implantation of cementless Zweymuller-stems-- A cross-sectional study]. AB - AIM: According to the results of longitudinal studies, the use of distally anchored femoral prostheses leads to marked proximal stress shielding. Zweymuller stems should be controlled some years after implantation. METHOD: In a cross sectional study, we examined 92 patients clinically, radiologically, and osteodensitometrically 2, 4, and 6 years after implantation in order to assess the osseointegration of Zweymuller shafts as independently as possible of age. The non-operated contralateral limb or Gruen's zone 4 below the prosthesis was used as a control group. RESULTS: At no time did the parameters assessed indicate any reduction in proximal bone density as compared with the control group. Zone 7 (medial-proximal) even showed an increase in bone density after 2 and 4 years. Nevertheless, absolute bone density was lowest proximally both pre- and postoperatively. Six years after implantation women even showed a 15 % increase in summated periprosthetic bone density as compared with the control group. No influence of femoral shape on bony remodeling was demonstrated. CONCLUSION: Periprosthetic bony remodeling cannot be attributed exclusively to the shape of the shaft. Longitudinal studies do not adequately reflect the reduction in periprosthetic bone mass that occurs as a function of age and altered activity level. The osseointegration of hip prostheses is influenced by other factors such as musculoskeletal lines of force, sex, and age. PMID- 12085300 TI - [Cortical strut grafts, an alternative to conventional plating in periprosthetic fractures of the femur]. AB - AIM: In periprosthetic fracture therapy plating has a failure rate between 30 to 50 %. Cortical allografts proved to unite constantly in revision surgery of the femoral shaft. The hypotheses to be answered in this study was whether the use of cortical allografts increases the success rate in contrast to plating. METHOD: A retrospective case control study has been done analysing the results in periprosthetic fractures of the femur around or below the tip of a stable femoral component (Vancouver B1, Mont III + IV). RESULTS: In 12 plating cases an average of 2.67 units of blood was transfused (0 - 6). In the follow-up two refractures, one loosening of the femoral component and one varus deformity greater than 10 degrees occurred. In 6 patients a total of 9 reoperations had to be done. According to the classification of Mont the results were excellent in 50 %, good in 16.7 % and poor in 33.3 %. 7 patients with strut grafts received an average of 1,43 units of blood transfused (0 - 3). In one patient with a 16 degrees varus deformity of the femur, due to multiple previous operations, a deformity of 21 degrees had to be accepted. Until now, no further operation had to be done in this group. The results were classified as excellent in 5 cases (71.4 %) and good in the other 2 (28,6 %). There was no poor result. The difference between the groups was statistically significant in the number of reoperations (p < 0. 05). CONCLUSION: In this study, with a low number so far, the strut graft group reached better results in every single variable, with a significant reduction of reoperations. PMID- 12085301 TI - [Functional postoperative treatment of internally fixed ankle fractures with a flexible arthrodesis boot (Variostabil)]. AB - AIM: Postoperative treatment following osteosynthesis of ankle fractures in a flexible arthrodesis boot (Variostabil) aims at fast restoration of the function of the injured extremity while allowing early full weight bearing. This treatment regimen was validated in a clinical and experimental study. METHODS: (1) In the clinical study part, 56 patients with internally fixed ankle fractures received after treatment with the arthrodesis boot for 6 weeks. (2) In the experimental study part, the intravascular pressure was recorded in a foot vein of 8 healthy volunteers during knee bends. RESULTS: (1) No implant failure or secondary dislocation was seen due to the after treatment. All patients rated subjective comfort and mobility as excellent. 90.5 % had a good to excellent functional result with the Philips Score. (2) Wearing the arthrodesis boot effected significantly faster venous outflow (25.8 +/- 15.2 vs. 11.3 +/- 6.0 mmHg/sec, p < 0.05) and higher pressure amplitude (53.6 +/- 12.0 vs. 26.5 +/- 9.6 mmHg) during knee bends, compared to a below-the-knee plaster cast. CONCLUSIONS: The flexible arthrodesis boot offers safe protection of ankle fractures combined with superior functional performance (undisturbed gait, training of the ankle joint, high patient comfort and mobility, accelerated venous outflow) as compared to cast immobilization. PMID- 12085303 TI - [Brace effect in scoliosis in the sagittal plane - an MRI study]. AB - AIM: Using magnetic resonance (MR) imaging we studied the brace effect in scoliosis in the sagittal plane. METHOD: In 38 patients with idiopathic scoliosis (mean age 13.4 years) MR total spine imaging was carried out to investigate the immediate effect of bracing in the sagittal plane. There were 19 thoracic, 13 S shaped and 6 lumbar scoliosis. On conventional radiographs the mean Cobb angle of the thoracic curves was 31 degrees and of the lumbar curves 26 degrees. MR imaging was performed in the supine position with and without the brace in direct sequence. On the sagittal MR projection the Cobb angle was measured between T 4 and T 12 and between T 12 and L 5. RESULTS: On the coronal MR images the mean correction with brace was 23 % of the thoracic curves and 29 % of the lumbar curves. The mean, sagittal Cobb angle (T 4 - T 12) was 14 degrees without brace and 12 degrees with brace. For the lumbar curves the mean sagittal Cobb angle (T 12 - L 5) was 32 degrees without brace and 31 degrees with brace. In the paired t test these differences were significant. CONCLUSION: Using MR total spine imaging the brace effect in scoliosis could be depicted in the sagittal plane. In the thoracic spine a correction of the lordotic deformity could not be observed. PMID- 12085302 TI - [CT morphometry for calcaneal fractures and comparison of the Zwipp and Sanders classifications]. AB - AIM: The aim of the study is to correlate the CT-morphological changes of fractured calcaneus and the classifications of Zwipp and Sanders with the clinical outcome. METHOD: In a retrospective clinical study, the preoperative CT scans of 75 calcaneal fractures were analysed. The morphometry of the fractures was determined by measuring height, length diameter and calcaneo-cuboidal angle in comparison to the intact contralateral side. At a mean of 38 months after trauma 44 patients were clinically followed-up. The data of CT image morphometry were correlated with the severity of fracture classified by Zwipp or Sanders as well as with the functional outcome. RESULTS: There was a good correlation between the fracture classifications and the morphometric data. Both fracture classifying systems have a predictive impact for functional outcome. The more exacting and accurate Zwipp classification considers the most important cofactors like involvement of the calcaneo-cuboidal joint, soft tissue damage, additional fractures etc. The Sanders classification is easier to use during clinical routine. CONCLUSION: The Zwipp classification includes more relevant cofactors (fracture of the calcaneo-cuboidal-joint, soft tissue swelling, etc.) and presents a higher correlation to the choice of therapy. Both classification systems present a prognostic impact concerning the clinical outcome. PMID- 12085304 TI - [Ultrasound in the early diagnosis of congenital dislocation of the patella]. AB - INTRODUCTION: Congenital dislocation of the patella is defined as lateral dislocation of the patella present at birth, impossibility of closed reduction and diagnosis before the age of 10 years. We report about a rare case of a bilateral congenital dislocation of the patella. CASE REPORT: Physical examination of an eight-month- old boy showed bilateral knee flexion contractures associated with moderate genu varum. On both sides the patella could not be palpated easily. Radiographic diagnosis could not show the patella as the patella normally ossifies later. Ultrasound examination located the patella lateral to the lateral femoral condyle on both sides. Closed reduction was impossible. An open reduction with division of the lateral soft tissues, lateral release, and derotation of the quadriceps femoris and refining of the medial structures was performed. After cast removal the patellae were both located in the intercondylar grooves as confirmed by clinical and ultrasound examinations of both knees. Active and passive exercises were started. CONCLUSION: Failure of internal rotation of the myotome which contains the quadriceps femoris and the patella is the etiology of congenital dislocation of the patella. The quadriceps acts as flexor, exerting a valgus stress on the knee, causing external rotation of the tibia. Diagnosis is often delayed because of the lack of pathological findings on plain radiographs. Early diagnosis is enabled by ultrasonography. Surgical treatment is necessary and results are good, as long as there are no secondary changes. PMID- 12085306 TI - [Influence of impaired glucose tolerance on long-term survival after acute myocardial infarction]. AB - BACKGROUND AND OBJECTIVE: An impaired glucose tolerance as well as type 2 diabetes are associated with an increased cardiovascular mortality. This study was undertaken to determine whether this is also true for impaired glucose tolerance in immediate temporal relation with an acute myocardial infarction (AMI). PATIENTS AND METHODS: The survival of 562 patients (232 females, 330 males; age 68 +/- 13 years) consecutively admitted to the intensive care unit of a medical department with the diagnosis of AMI, were prospectively evaluated over a span of more than 3 years. Type 2 diabetes had been previously known in 152, while it was newly diagnosed in 83 patients. Oral glucose tolerance tests (OGTT) were performed in 129, but the test was not performed in 222 patients. Survival was analysed with the Kaplan-Meier test. RESULTS: Among the 129 patients who had had an OGTT, it was normal in 60 (47%), 45 (35%) had impaired glucose tolerance and 24 (19%) were found to have previously undiagnosed diabetes (WHO criteria). Survival of all patients with type 2-diabetes was significantly worse than in the remainder of patients (p < 0.0001). In patients with impaired glucose tolerance the survival time was significantly shorter than in those with normal glucose tolerance (p = 0.029). Even after excluding those patients who had died in the acute post-infarction phase, the difference between patients with normal and with impaired glucose tolerance remained significant (p = 0.034). CONCLUSION: Patients with impaired glucose tolerance have a significantly shorter survival after AMI than those with a normal glucose tolerance. Blood glucose concentration 2 hours after oral glucose intake thus seems to be an important predictor of death after AMI. PMID- 12085305 TI - [Interposition-arthroplasty of the elbow in a child as delayed treatment of an unreduced multifragmentary fracture of the distal humerus: result after 77 years- case report and review of the literature]. AB - Post-traumatic stiffness of the elbow joint in children after unreduced multifragmentary fractures of the distal humerus can be successfully treated by interposition arthroplasty. A case with long-term follow up is described, and the literature discussed. PMID- 12085307 TI - [Recurrent lower gastro-intestinal bleeding in a 38-year-old patient]. AB - HISTORY AND CLINICAL FINDINGS: A 38-year-old patient with a history of recurrent gastrointestinal bleeding for more than 15 years was transferred for the treatment of a new onset of hematochezia and anemia. His general condition was clearly reduced. EXAMINATIONS: Laboratory results showed a distinct microcytic hypochromic anemia; ferrum, ferritin, albumin, and protein were also diminished. Colonoscopy and transrectal ultrasound uncovered large hemangiomatous structures in the rectum. Digital substraction angiography of the bilateral internal iliac artery, superior and inferior mesenteric arteries including its branches showed a perirectal polypoid tumour with a discrete angiomatous structural shadow in the capillary phase. MRI of the pelvis revealed angiomatous blood vessels in the whole pararectal space and to a minor extension in the gluteal muscles. DIAGNOSIS, TREATMENT AND COURSE: The diagnosis of cavernous hemangioma of the rectum was made and a distal mesorectal resection of the rectum including a coloanal anastomosis and double-running ileostomy was performed. 3 months later the ileostomy was reversed. There was no further bleeding and the rectal continence was normal. CONCLUSION: In case of unclear recurrent lower gastrointestinal bleeding, with onset in the early childhood and impressive rectal varices, one has to think about the rare diagnosis of cavernous hemangioma. PMID- 12085309 TI - [Chronic heart failure--treatment]. PMID- 12085310 TI - [Mental changes in hypo- and hyperthyroidism]. PMID- 12085311 TI - [The influence of genetics on health-insurance]. PMID- 12085308 TI - [Coronary-subclavian steal syndrome]. AB - HISTORY AND CLINICAL FINDINGS: A-46-year-old woman with generalized atherosclerosis and coronary triple vessel disease was admitted with recurrent angina pectoris 17 months after primarily successful triple coronary artery bypass grafting (CABG). The symptoms were induced by exercise of the arms, e. g. carrying shopping bags in the left hand. INVESTIGATIONS: Stress technetium scans showed pronounced left ventricular ischemia of the anterior wall and apex during left upper limb exercise in comparison to bicycle exercise. The coronary angiography showed reverse flow in the internal mammary artery (IMA) graft to the left anterior descending artery (LAD), and aortography disclosed blockage of the proximal left subclavian artery. TREATMENT AND COURSE: : The patient declined percutaneous transluminal interventions or surgical procedures of the left subclavian artery. Because of mild angina and the lack of neurological symptoms, she was discharged on optimal drug treatment. CONCLUSION: : Patients with recurrence of cardiac symptoms after CABG surgery and IMA graft may have a coronary-subclavian steal syndrome. Clinically, these patients present with arterial blood pressure differences between left and right arm of typically more than 20 mmHg. The therapy of choice are percutaneous transluminal revascularization procedures of the subclavian artery. PMID- 12085312 TI - [Japanese encephalitis]. PMID- 12085313 TI - Dengue-specific T cell responses in peripheral blood mononuclear cells obtained prior to secondary dengue virus infections in Thai schoolchildren. AB - Children who experience secondary dengue virus (DV) infections are at increased risk for dengue hemorrhagic fever. To study the effect of preexisting T cell responses to DV on the severity of secondary virus infection, peripheral blood mononuclear cells (PBMC) from 10 subsequently hospitalized and 12 nonhospitalized Thai schoolchildren were stimulated with inactivated dengue antigens, and proliferation of interferon (IFN)-gamma or tumor necrosis factor (TNF)-alpha responses of the preinfection PBMC were measured. Proliferation responses were observed in 11 subjects, and IFN-gamma responses were seen in 12 subjects, 6 of whom showed broad serotype cross-reactive IFN-gamma responses. TNF-alpha responses were detected exclusively in 4 hospitalized subjects. Four PBMC samples that showed neither proliferation nor cytokine responses to any dengue antigen were from nonhospitalized subjects. This study, thought to be the first to investigate T cell responses to DV in preinfection PBMC, suggests that the pattern of preexisting T cell responses influences the risk for severe disease. PMID- 12085314 TI - Chemokine-induced leishmanicidal activity in murine macrophages via the generation of nitric oxide. AB - This study explored the role of the proinflammatory chemokines macrophage inflammatory protein (MIP)-1alpha and macrophage chemoattractant protein (MCP)-1 for development of antileishmanial activity. There was substantial inhibition in nitrite generation in Leishmania donovani-infected macrophages. A marked elevation of nitrite generation and induction of inducible nitric oxide (NO) synthase (iNOS) mRNA was found in chemokine-primed parasite-infected macrophages. Tumor necrosis factor-alpha, which is the priming signal for NO production, was also up-regulated under similar experimental conditions. The priming with chemokine inhibited the multiplication of L. donovani amastigotes within the intramacrophageal milieu. The antileishmanial effect of chemokines was almost completely abrogated when the macrophages were preincubated with l-N-monomethyl arginine, the specific inhibitor of iNOS. The results of this investigation suggest that the CC chemokines MIP-1alpha and MCP-1 orchestrate an antileishmanial armamentarium via the induction of an NO-mediated regulatory mechanism to control the intracellular growth and multiplication of the Leishmania protozoan. PMID- 12085315 TI - Cytomegalovirus (CMV) phosphoprotein 65 makes a large contribution to shaping the T cell repertoire in CMV-exposed individuals. AB - Antigen-specific, cytokine flow cytometry was used to analyze the prevalence and frequency of CD4 and CD8 memory T cells specific for the abundantly expressed cytomegalovirus (CMV) phosphoprotein 65 (pp65) in healthy CMV IgG-seropositive individuals. Stimulation of peripheral blood mononuclear cells with peptide pools and individual peptides derived from the pp65 amino acid sequence in 40 donors revealed that 63% of donors had a detectable CD4 T cell response and that 83% of donors had a detectable CD8 T cell response against this protein. The overall frequencies of T cells directed against pp65 were analyzed for 20 donors by stimulation with peptide pools covering the complete pp65 protein and were as high as 2 in 1000 and 9 in 1000 (median) peripheral blood CD4 and CD8 T cells, respectively. In addition, a comparison between CD4 responses to a CMV lysate containing various CMV proteins and pp65-specific responses in 9 donors indicated that pp65 was a dominant target of the CMV-specific CD4 T cell response in some, but not all, donors. Several new T cell epitopes were identified. PMID- 12085317 TI - Human cytomegalovirus (HCMV) replication dynamics in HCMV-naive and -experienced immunocompromised hosts. AB - Human cytomegalovirus (HCMV) can infect both HCMV-naive and -experienced transplant patients. In this study, the growth rate of HCMV in HCMV-naive hosts (1.82 units/day; 95% confidence interval [CI], 1.44-2.56 units/day) was shown to be significantly faster than the growth rate of virus in HCMV-experienced hosts undergoing recurrent infection (0.61 units/day; 95% CI, 0.55-0.7 units/day; P<.0001). The basic reproductive number (R(0)) for HCMV-naive liver transplant patients was 15.1 (95% CI, 8.9-44) but was only 2.4 (95% CI, 2.35-2.8) for HCMV experienced transplant recipients, corresponding to an anti-HCMV immune efficacy of approximately 84%, despite immunosuppressive therapy. The R(0) values suggest that an anti-HCMV drug or vaccine with an efficacy of >93% (95% CI, 89%-98%) is required to eliminate viral growth during infection of HCMV-naive liver transplant recipients, whereas lower efficacy levels are sufficient to reduce the R(0) value to <1 in hosts with prior HCMV immunity. PMID- 12085316 TI - Cytomegalovirus (CMV) polymerase chain reaction profiles in individuals with advanced human immunodeficiency virus infection: relationship to CMV disease. AB - Cytomegalovirus (CMV) disease is a common complication of patients with advanced human immunodeficiency virus infection. The aim of the present study, based on a case-cohort design, was to determine the predictive value of follow-up and baseline qualitative plasma CMV polymerase chain reaction (PCR) values for CMV end-organ disease in 378 patients (158 who progressed to CMV end-organ disease and 220 who did not develop CMV disease). These patients are part of the full AIDS Clinical Trials Group 204 multinational study (1227 patients), a randomized, controlled trial that compared the effects of valacyclovir with those of acyclovir for CMV disease prevention. Baseline PCR positivity was a significant risk factor for CMV disease progression (relative risk [RR], 1.81; 95% confidence interval [CI], 1.09-3.00). In multivariate analyses, time-updated PCR positivity was strongly associated with progression to CMV end-organ disease (RR, 4.42; 95% CI, 2.87-6.81). Change in cumulative PCR status was informative for the risk of subsequent CMV disease. PMID- 12085318 TI - Discordance at human leukocyte antigen-DRB3 and protection from human immunodeficiency virus type 1 transmission. AB - Host human leukocyte antigens (HLAs) integrated into the human immunodeficiency virus (HIV) type 1 envelope could theoretically determine, as in tissue transplants, whether HIV-1 is "rejected" by exposed susceptible persons, preventing transmission. HLA discordance (mismatch) was examined among 45 heterosexual partner pairs in which at least 1 partner was HIV-1 infected and exposure or transmission between partners had occurred. Immunologic discordance at class II HLA-DRB3 (present in the HIV donor partner but absent in the recipient partner) was associated with lack of transmission of HIV-1. Eight (35%) of 23 partner pairs in which HIV-1 transmission did not occur were immunologically discordant at HLA-DRB3, compared with 0 of 11 partner pairs in which HIV-1 transmission did occur (P=.027). Further investigation of the roles of class II HLAs in HIV-1 transmission and as possible components of HIV-1 vaccines should be pursued. PMID- 12085319 TI - Elevated virus loads of Kaposi's sarcoma-associated human herpesvirus 8 predict Kaposi's sarcoma disease progression, but elevated levels of human immunodeficiency virus type 1 do not. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is found in Kaposi's sarcoma (KS), multicentric Castleman's disease, and primary effusion lymphomas. To prospectively evaluate KSHV load as a biomarker for KS clinical status and prognosis in a cohort of men with AIDS-related KS, 2 quantitative polymerase chain reaction (PCR) assays were developed and tested to determine KSHV peripheral blood mononuclear cell (PBMC) virus loads. Most patients (13/15) with good-prognosis KS had < or =1.5 log KSHV copies/10(5) PBMC by both quantitative competitive (QC) and real-time Applied Biosystems (ABI) PCR. Both assays provided 94% specificity for identifying the 16 patients without KS progression during 20 months of follow-up. QC-PCR and ABI-PCR exhibited 100% and 80% levels of diagnostic sensitivity, respectively, for identifying the 5 patients whose KS progressed. Neither dichotomized human immunodeficiency virus loads nor dichotomized CD4 counts predicted either KS progression or KS clinical stage (all positive predictive values < 30%). These results are evidence that the quantity of circulating KSHV in KS patients is biologically meaningful and is measurable with sufficient accuracy to provide clinically useful information. PMID- 12085320 TI - Group B streptococcal beta-hemolysin induces mortality and liver injury in experimental sepsis. AB - New Zealand White rabbits were challenged with the wild-type (wt) group B streptococci (GBS) serotype III strain (COH1) and its isogenic nonhemolytic (NH) and hyperhemolytic (HH) mutants. Mortality differed significantly between rabbits infected with the HH mutant IN40 (67%), compared with rabbits infected with the wt COH1 strain (27%) and the NH strains COH1-20 and COH1:cylEDeltacat (13% and 0%, respectively; P<.05). Histopathologically, disseminated septic microabscesses surrounded by necrotic foci were found exclusively in the livers of HH mutant IN40-infected animals. Serum transaminase levels were 20-fold higher in the HH infected group, compared with rabbits infected with the other strains. Positive TUNEL (in situ terminal deoxynucleotide transferase-mediated dUTP nick end labeling) staining and activation of caspase-3 in hepatocytes were more frequent in HH-infected than in wt-infected animals and absent in the NH mutant COH1-20 infected group, indicating that GBS beta-hemolysin triggers apoptotic pathways in hepatocytes. This work provides the first evidence that GBS beta-hemolysin plays a crucial role in the pathophysiology of GBS sepsis by inducing liver failure and high mortality. PMID- 12085321 TI - Humanlike immune response of human leukocyte antigen-DR3 transgenic mice to staphylococcal enterotoxins: a novel model for superantigen vaccines. AB - This study examined the biologic responses of transgenic mice expressing human leukocyte antigen (HLA)-DR3 and human CD4 molecules, in the absence of murine major histocompatibility complex (MHC) class II molecules (Ab(0)), to staphylococcal enterotoxins (SEs) and evaluated protective immunity of a nonsuperantigen form of SEB against wild-type holotoxin. HLA-DR3 transgenic mice responded to several log lower concentrations of SEs and secreted higher levels of proinflammatory cytokines than did wild-type mice. Vaccination of transgenic mice with a nonsuperantigenic form of SEB induced high levels of neutralizing anti-SEB antibodies, which protected the mice from a surge in proinflammatory cytokine secretion after SEB challenge. The humanlike responses of the transgenic mice to SEs support the hypothesis that these mice represent an appropriate model to examine vaccines and therapeutics against SEs. This is thought to be the first report of examination of a vaccine against SEB in the context of human MHC class II receptors. PMID- 12085322 TI - Prospective study of infective endocarditis among injection drug users. AB - To determine the effect of human immunodeficiency virus (HIV) infection and other factors on infective endocarditis (IE) among injection drug users (IDUs), the incidence of IE was determined according to HIV status in a cohort of IDUs. A nested case-control study assessed IE risk factors. IE incidence (117 cases) was higher among HIV-seropositive than HIV-seronegative IDUs (13.8 vs. 3.3 cases/1000 person-years) during 1988-1998. Multivariate analysis of HIV-infected case patients revealed an inverse association between IE and CD4 lymphocyte count (odds ratio [OR] for 200-499 cells/mm(3), 2.01; OR for <200 cells/mm(3), 3.61) and with alcohol intake (OR for 1-21 drinks/week, 0.43; OR for >21 drinks/week, 0.32). Women had an increased risk of IE (OR, 3.26), as did persons with increasing injection drug use frequency (OR for less than daily use, 3.15; OR for at least daily use, 6.07). This study confirms that IE is more common among IDUs with advanced HIV immunosuppression even after accounting for injection drug use behaviors. PMID- 12085323 TI - Bactericidal group IIA phospholipase A2 in serum of patients with bacterial infections. AB - Group IIA phospholipase A2 (PLA2-IIA) is a newly recognized antibacterial acute phase protein. The concentration of PLA2-IIA increases up to 500-fold in the blood plasma of patients with severe acute diseases, compared with healthy persons. Despite numerous studies, the exact roles of this enzyme in human diseases are unknown. This study investigated the antibacterial properties of PLA2-IIA in human acute phase serum. PLA2-IIA in serum samples of patients with bacterial infections was capable of killing 90% of Staphylococcus aureus and 99% of Listeria monocytogenes in vitro after incubation for 2 h. At concentrations found in normal human serum, PLA2-IIA killed 90% of L. monocytogenes but did not kill S. aureus or Escherichia coli. The bactericidal effects of acute phase and normal human serum were abolished after depletion of PLA2-IIA by immunoadsorption. PMID- 12085325 TI - Cytokine and chemokine gene expression after primary and secondary respiratory syncytial virus infection in cotton rats. AB - The induction of pro- and anti-inflammatory cytokines and chemokines was studied in the lungs of cotton rats after primary or secondary infection with respiratory syncytial virus (RSV). Increases in messenger RNA (mRNA) levels of all genes analyzed were observed during the course of primary infection. In general, mRNA expression peaked between postinfection days 1 and 4 and returned to near-normal levels by day 10. During secondary infection, the expression of some genes (i.e., interferon [IFN]-gamma and interleukin [IL]-10) began earlier, some (i.e., IL 1beta and macrophage inflammatory protein-1beta) began later, and some (i.e., IL 1beta, IL-10, growth-regulated protein, and tumor necrosis factor-alpha) showed prolonged expression, whereas 2 genes (i.e., IFN-alpha and IL-6) were not expressed. This study presents evidence of different kinetics of expression of inflammatory mediators during primary and secondary infection that likely coincide with innate and adaptive immune response and complement previous observations that emphasize the role of inflammation in the pathogenesis of RSV disease. PMID- 12085324 TI - Human phagocytic cells in the early innate immune response to Borrelia burgdorferi. AB - During natural infection with the agent of Lyme disease, Borrelia burgdorferi, polymorphonuclear leukocytes (PMNL) are the first cells of the innate immune system to arrive at the site of spirochete deposition in the skin. This study examined the degree of spirochete clearance likely to occur with PMNL or mononuclear cells before the development of the secondary immune response. Without specific antibody in vitro, there was very limited uptake of spirochetes by PMNL or monocytes and no intracellular colocalization of PMNL granule products with spirochetes. Most of the killing of spirochetes by PMNL was extracellular. In contrast, mature macrophages ingest and kill spirochetes avidly with or without specific antibody. Once the spirochetes are opsonized, PMNL clear them rapidly. These findings may be relevant to the initial survival of spirochetes introduced into the host. PMID- 12085326 TI - Onchocerca volvulus microfilariae avoid complement attack by direct binding of factor H. AB - The filarial parasite Onchocerca volvulus is the causative agent of river blindness. The adult worms produce microfilariae (mf), which are responsible for the disease pathogenesis; mf activate the complement system, but the activation stops before the formation of terminal complement complexes. Because of the arrest of complement activation, this study analyzed binding of the main alternative pathway regulator, factor H (fH), to the mf. The mf bound fH after incubation in nonimmune human serum or with purified radiolabeled fH. In the presence of factor I, mf-bound fH promoted the cleavage of complement 3 molecule b (C3b) to iC3b. An analysis with recombinant constructs of fH showed that the C terminal short consensus repeats (SCRs) 8-20 of fH bound to mf, whereas the N terminal SCRs 1-7 containing the complement-regulatory domains in SCRs 1-5 did not. Thus, mf of the nematode O. volvulus may evade human complement by binding fH and by promoting inactivation of C3b into iC3b. PMID- 12085327 TI - Identification and characterization of 3 novel genital human papillomaviruses by overlapping polymerase chain reaction: candHPV89, candHPV90, and candHPV91. AB - Three novel human papillomaviruses (HPVs), candHPV89, candHPV90, and candHPV91, that were previously identified from short polymerase chain reaction (PCR) fragments AE6/CP6108, JC9710, and JC9813, respectively, were cloned and characterized from cervicovaginal cells by use of an overlapping PCR method. The complete nucleotide sequences of candHPV89 (8078 bp), candHPV90 (8033 bp), and candHPV91 (7966 bp) were determined by sequence walking. candHPV89 and candHPV91 were closely related to HPV83 and HPV7 and were placed in the HPV genome homology groups A3 and A8, respectively, by phylogenetic analyses. The genome of candHPV90 was most closely related to HPV71, although these HPV genomes do not seem to form a single lineage, because of the disproportionate divergence of the HPV71 L1 region. On the basis of phylogenetic analyses and available clinical data, these 3 novel HPV genomes appear to have a low oncogenic risk and expand the heterogeneity of HPVs detected in the lower genital tract. PMID- 12085328 TI - Evaluation of coxsackievirus infection in children with human immunodeficiency virus type 1-associated cardiomyopathy. AB - In a matched case-control study of the association between coxsackieviruses and cardiac impairment, 24 human immunodeficiency virus (HIV) type 1-infected children with cardiac impairment were compared with 24 HIV-1-infected control subjects. Serologic evidence of coxsackievirus infection was present in all children, with no significant difference in geometric mean antibody titers between case patients and control subjects. Conditional logistic regression to test for an association between coxsackievirus antibody titer and the presence or absence of cardiac impairment, by any indicator, showed an odds ratio of 1.11 (95% confidence interval, 0.58-2.10; P=.75), indicating no association between coxsackievirus infection and cardiac impairment. Coxsackievirus antibody titers correlated positively with total IgG levels in nonrapid progressors but not in rapid progressors. Paired serum samples taken before and after diagnosis of cardiac impairment in 5 patients showed no evidence of intervening coxsackievirus infection. These results do not identify a causal role for coxsackieviruses for cardiomyopathy in HIV-1-infected children. PMID- 12085329 TI - Reduced CC chemokine receptor (CCR) 1 and CCR5 surface expression on peripheral blood T lymphocytes from patients with chronic hepatitis C infection. AB - T cell recruitment to the infected liver is an essential step for the efficient elimination of hepatitis viruses. The surface expression of CC chemokine receptor (CCR) 1, CCR4, and CCR5 on peripheral blood T lymphocytes and their responsiveness to the chemokines macrophage inflammatory proteins (MIP)-1alpha, MIP-1beta, and RANTES (regulated on activation, normally T cell-expressed and secreted) was analyzed in patients with chronic hepatitis C and hepatitis B infection and compared with healthy subjects. Although CCR4 surface expression was not altered, hepatitis C virus (HCV)-infected patients had lower proportions of CD8 T cells with CCR1 and CCR5 surface expression (P<.05). Migration of CD8 T cells in response to MIP-1alpha, MIP-1beta, and RANTES was significantly reduced in HCV-infected patients (P<.05). Intracellular CCR1 and CCR5 protein and messenger RNA levels in peripheral blood T cells did not indicate reduced chemokine receptor biosynthesis in hepatitis C infection. Thus, chronic hepatitis C, but not hepatitis B, infection alters surface expression of distinct CCRs, resulting in lower CC chemokine responsiveness. PMID- 12085330 TI - Risk factors for human herpesvirus 8 infection in a cohort of drug users in the Netherlands, 1985-1996. AB - To elucidate the mode of human herpesvirus 8 (HHV-8) transmission in a population of Amsterdam drug users, HHV-8 seroprevalence and seroincidence were determined in 1179 drug users in the Amsterdam Cohort Studies (1985-1996). Risk factors for HHV-8 infection were examined. Serum samples were screened with an enzyme immunoassay by using HHV-8 lytic capsid (open-reading frame [ORF] 65) and latent nuclear (ORF73) antigens; positive results were confirmed by Western blot and immunofluorescence assay. Seroprevalence (men, 3.4%; women, 1.4%) and seroincidence (men, 0.08; women, 0.05/100 person-years) were low in this study. Infections with human immunodeficiency virus (HIV) type 1, hepatitis B virus (HBV), and hepatitis C virus (HCV), but not HHV-8, were associated with injection drug use (IDU). Independent risk factors for HHV-8 seropositivity were homosexual contacts and Mediterranean nationality for men and sexual contact with bisexual men, absence of a steady partner, and unprotected commercial sex for women. Unlike HIV-1, HBV, or HCV infection, HHV-8 infection is uncommon in Amsterdam drug users, as is HHV-8 transmission through IDU. PMID- 12085331 TI - Cytomegalovirus (CMV) retinitis immune restoration disease occurs during highly active antiretroviral therapy-induced restoration of CMV-specific immune responses within a predominant Th2 cytokine environment. AB - Plasma levels of cytomegalovirus (CMV)-specific immunoglobulin G (IgG), soluble (s) CD30, sCD26 (dipeptidyl peptidase IV [DPP IV]) enzyme activity, and tumor necrosis factor receptor-I (TNFR-I) were assessed in human immunodeficiency virus (HIV)-infected patients who experienced CMV retinitis (CMVR) as an immune restoration disease (IRD) during their first 6 months of highly active antiretroviral therapy (HAART) and in CMV-seropositive, HIV-infected patients with similar baseline CD4(+) T cell counts who had uneventful immune reconstitution. Patients who experienced CMVR IRD had a significant increase in CMV-specific IgG during their first 12 months of HAART, indicating restored CMV specific immune responses. They also had significantly higher levels of sCD30 both before HAART and for up to 12 months after start of treatment. sCD30 levels remained elevated during 48 months of HAART, suggesting persistence of a predominant Th2 cytokine environment. Levels of sCD26 (DPP IV) enzyme activity and TNFR-I did not differ significantly between the 2 groups at any time point. PMID- 12085332 TI - Evaluation of initial CD4+ T cell counts in individuals with newly diagnosed human immunodeficiency virus infection, by sex and race, in urban settings. AB - The CD4+ T cell count is an important determinant of disease stage and prognosis in human immunodeficiency virus (HIV)-infected individuals. This study evaluated the CD4+ T cell counts in individuals at the time of diagnosis of HIV infection at 4 community clinics in large urban settings with relatively high frequencies of HIV infection. Of 2223 individuals, 57% and 36% had CD4+ T cell counts < 350 and < 200 cells/mm(3), respectively, at the time of diagnosis. There were no clear differences by sex or race. Enhanced educational efforts regarding the importance of HIV testing for at-risk individuals across sex and race strata in community settings may be important for early identification of individuals with HIV infection. This in turn could impact efforts to reduce transmission, and it could impact the prognosis for patients who receive antiretroviral therapy. PMID- 12085333 TI - A prospective study of hormonal contraceptive use and cervical shedding of herpes simplex virus in human immunodeficiency virus type 1-seropositive women. AB - Cross-sectional analyses have demonstrated an association between use of hormonal contraceptives and shedding of herpes simplex virus (HSV). This prospective study evaluated the effect of initiating use of hormonal contraception on cervical HSV detection. Two hundred women who were seropositive for HSV-2 and human immunodeficiency virus (HIV) type 1 were examined for cervical mucosal HSV by use of quantitative DNA polymerase chain reaction before and after beginning the use of hormonal contraceptives. Cervical HSV was detected in 32 women (16.0%) before initiating and in 25 women (12.5%) after initiating use of hormonal contraception (P=.4). There were no significant differences in HSV shedding among the subgroups of women starting combination oral contraceptives containing both estrogen and progesterone or progesterone-only contraceptives. Among the 54 women who shed HSV at least once, the median change in cervical HSV after initiation of hormonal contraception was -313 copies/swab. In this prospective study, use of hormonal contraceptives did not increase detection of cervical HSV. PMID- 12085334 TI - Morphine promotes simian acquired immunodeficiency syndrome virus replication in monkey peripheral mononuclear cells: induction of CC chemokine receptor 5 expression for virus entry. AB - Rhesus monkeys (Macaca mulatta) chronically administered opioids were more susceptible to simian immunodeficiency virus (SIV) strain mac239 (SIVmac239) infection than those without prior exposure to opioids. Increased plasma viremia in morphine-dependent monkeys allowed SIV to be detected in the animals' peripheral blood mononuclear cells (PBMC) without cocultivation with a tissue culture cell line. In contrast, virus titers from the PBMC of morphine-naive SIVmac239-infected animals were undetectable in the absence of cocultivation. PBMC isolated from noninfected animals and treated with morphine sulfate in vitro produced an increase in the expression of beta-chemokine receptor 5 (CCR5). Because both SIVmac239 and human immunodeficiency virus (HIV) require CCR5 for cell entry, the unique role of morphine in promoting SIV infection may provide a mechanism to account for the high incidence of HIV disease among drug-using populations. PMID- 12085335 TI - Antifungal therapy for central nervous system histoplasmosis, using a newly developed intracranial model of infection. AB - The outcome of central nervous system (CNS) histoplasmosis is often unfavorable. Although fluconazole plays an integral role in treatment of fungal meningitis, its role in the treatment of histoplasmosis is hampered by reduced activity and potential development of resistance. A murine model of CNS histoplasmosis was used to evaluate the hypothesis that a combination of amphotericin B and fluconazole therapy would be superior to amphotericin B monotherapy. Groups of B6C3F(1) mice were infected by injection of Histoplasma capsulatum into the subarachnoid space. The addition of fluconazole hindered the antifungal effect of amphotericin B, as determined by measurement of fungal burden, suggesting antagonism in the brain. Fluconazole was less effective as a single agent than was amphotericin B, despite the greater penetration of fluconazole into brain tissues. The hypothesis that amphotericin B-fluconazole combination therapy would be superior to amphotericin B monotherapy for treatment of CNS histoplasmosis was not supported by this study. PMID- 12085336 TI - Critical role of myeloperoxidase and nicotinamide adenine dinucleotide phosphate oxidase in high-burden systemic infection of mice with Candida albicans. AB - Oxygen metabolites generated by myeloperoxidase (MPO) and nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase contribute to microbial killing by phagocytes. To compare the importance of the 2 enzymes for host defense, MPO deficient (MPO(-/-)) mice and NADPH-oxidase-deficient mice with chronic granulomatous disease (CGD mice) were intraperitoneally infected with 3 different doses of Candida albicans, and their infection severity was analyzed. CGD mice had increased mortality and exhibited increased tissue fungal burden in a dose dependent manner, whereas normal mice showed no symptoms. Of interest, at the highest dose, the mortality of MPO(-/-) mice was comparable to that of CGD mice, but at the lowest dose, it was the same as that of normal mice. At the middle dose, the number of fungi disseminated into various organs of the MPO(-/-) mice was comparable to that of the CGD mice at day 6 of infection, but it was significantly lower at day 14. These results suggest that MPO and NADPH-oxidase are equally important for early host defense against a large inoculum of Candida. PMID- 12085337 TI - Dynamics of gametocytes among Plasmodium falciparum clones in natural infections in an area of highly seasonal transmission. AB - The dynamics of gametocyte production in Plasmodium falciparum clones were studied in inhabitants of an area of highly seasonal malaria transmission in eastern Sudan. Reverse-transcriptase polymerase chain reaction was used to detect expression of 2 genes that encode gametocyte-specific proteins, pfs25 and pfg377, in parasites sampled from individuals throughout one year. Some patients who acquired infections during the wet season were found to harbor subpatent gametocytemia through the following dry season in the apparent absence of mosquito transmission. Genotyping of parasites in multiclonal infections showed considerable fluctuation of gametocyte production by individual clones. The gametocytes present at the end of the dry season provide the most probable source of the genetically complex cyclical malaria outbreaks following the rainy season in this region. PMID- 12085338 TI - Reactivation of human herpesvirus 6 infection in an immunocompetent host with chronic encephalopathy. PMID- 12085340 TI - Impact of protease polymorphisms and viral fitness on human immunodeficiency virus (HIV) type 1 viremia in untreated HIV-1 infection. PMID- 12085341 TI - Identification of the cryptosporidium pig genotype in a human patient. PMID- 12085342 TI - Hepatitis C infection and the patient with end-stage renal disease. AB - Hepatitis C virus (HCV) remains common in patients with end-stage renal disease (ESRD) and is an important cause of liver disease in this population. Acquisition of HCV infection continues to occur in dialysis patients because of nosocomial spread. The natural history of HCV in dialysis patients remains controversial because the course of HCV typically extends over decades, whereas dialysis patients have higher morbidity and mortality rates than those of the general population limiting long-term follow-up. However, recent reports suggest that HCV infection affects the survival of chronic dialysis patients as well as renal transplant (RT) recipients. The severity of preexisting liver disease on pretransplantation liver biopsy may provide useful prognostic information about clinical outcome after RT; liver biopsy should be incorporated in the evaluation and management of RT candidates with HCV. Recent surveys with long-term follow-up have documented adverse effects of HCV on patient and graft survival. Use of renal grafts from HCV-infected donors in recipients with HCV does not appear to result in a greater burden of liver disease albeit with short-term follow-up. There is only limited data about interferon (IFN) therapy in chronic dialysis patients, although sustained responses are reported. Preliminary data on IFN plus ribavirin therapy in dialysis patients with hepatitis C have given encouraging results, but randomized trials are needed. Interferon remains contraindicated post-RT because of concern about precipitating graft dysfunction. PMID- 12085344 TI - Leptin is required for fibrogenic responses induced by thioacetamide in the murine liver. AB - In this study, we investigated hepatic fibrogenesis caused by long-term thioacetamide (TAA) administration in ob/ob mice, a naturally occurring leptin deficient animal. In the lean littermates, prominent hepatic fibrosis, as well as positive staining for alpha smooth muscle actin (alpha-SMA), was induced by treatment with TAA (200 microg/g, IP, 3 times per week) for 4 to 8 weeks as expected. In sharp contrast, almost no hepatic fibrosis developed in ob/ob mice given the equivalent doses of TAA, where specific staining for alpha-SMA barely was detected. Induction of alpha1(I) procollagen mRNA caused by TAA also was prevented in ob/ob mice almost completely. Further, transforming growth factor beta (TGF-beta) mRNA was increased in the liver after TAA treatment for 4 weeks in lean littermates, which also was prevented in ob/ob mice. Interestingly, fibrotic septa in the hepatic lobules, as well as increases in alpha1(I) procollagen mRNA, was observed in ob/ob mice, when they were injected with recombinant murine leptin (1 microg/g daily) in combination with TAA treatment. Leptin per se did not cause any fibrotic changes in the liver in ob/ob mice. These findings clearly indicated that leptin deficiency is responsible for the resistance to TAA-induced profibrogenic responses in ob/ob mice. In conclusion, leptin appears to promote profibrogenic responses in the liver, in part, by up regulation of TGF-beta. PMID- 12085345 TI - Embryoid-body cells derived from a mouse embryonic stem cell line show differentiation into functional hepatocytes. AB - Embryonic stem (ES) cells have a potential to differentiate into various progenitor cells. Here we investigated the differentiation capacity of mouse ES cells into hepatocytes both in vitro and in vivo. During the culture of embryoid bodies (EBs) derived from ES cells, albumin (ALB) messenger RNA (mRNA) was expressed within 12 days after removal of leukemia inhibitory factor, and alpha fetoprotein (AFP) mRNA was observed within 9 days without additional exogenous growth factors. In ES cells and early EBs, by contrast, neither ALB mRNA nor AFP mRNA was observed. ALB protein was first detected at day 15 and the level increased with the culture period. The differentiation of EBs facilitated the synthesis of urea with the culture period, whereas early EBs and ES cells produced no urea. These results suggest that cultured EBs contain hepatocytes capable of producing ALB and urea. ES cells and the isolated cells from EBs were transplanted through portal vein to the liver after 30% partial hepatectomy of female mice pretreated with 2-acetylaminofluorene. Four weeks after transplantation with isolated cells from day-9 EBs, ES-derived cells containing Y chromosome in the liver were positive for ALB (0.2% of total liver cells), whereas teratoma was found in mice transplanted with ES cells or EBs up to day 6. The incidence of teratoma was decreased with the culture duration and no teratoma was observed in the liver transplanted with isolated cells from day-9 EBs. In conclusion, our in vitro and in vivo experiments revealed that cultured EBs contain functional hepatocytes or hepatocyte-like cells. PMID- 12085346 TI - Differential regulation of cyclins D1 and D3 in hepatocyte proliferation. AB - Substantial evidence suggests that cyclin D1 plays a pivotal role in the control of the hepatocyte cell cycle in response to mitogenic stimuli, whereas the closely related protein cyclin D3 has not been extensively evaluated. In the current study, we examined the regulation of cyclins D1 and D3 during hepatocyte proliferation in vivo after 70% partial hepatectomy (PH) and in culture. In contrast to cyclin D1, which was nearly undetectable in quiescent liver and substantially up-regulated after PH, cyclin D3 was constitutively expressed and induced only modestly. In the regenerating liver, the concentration of cyclin D3 was only about 10% of that of cyclin D1. Cyclin D1 formed complexes primarily with cyclin-dependent kinase 4 (cdk4), which were markedly activated in the regenerating liver and readily sequestered the cell cycle inhibitory proteins, p21 and p27. Cyclin D3 bound to both cdk4 and cdk6. Cyclin D3/cdk6 activity was readily detectable in quiescent liver and changed little after PH, and this complex appeared to play a minor role in sequestering p21 and p27. In cultured hepatocytes, epidermal growth factor or insulin had little effect, but the combination of these agents substantially induced cyclin D1 and cell cycle progression. Inhibition of Mek1 or phosphoinositide 3-kinase markedly inhibited cyclin D1 expression and replication. In contrast, cyclin D3 was expressed in the absence of mitogens and was only modestly affected by these manipulations. In addition, growth-inhibitory extracellular matrix conditions inhibited cyclin D1 but not cyclin D3 expression. In conclusion, these results support the concept that cyclin D1 is critically regulated by extracellular stimuli that control proliferation, whereas cyclin D3 is regulated through different pathways and plays a distinct role in the liver. PMID- 12085347 TI - Cyclin kinase inhibitor p21 potentiates bile acid-induced apoptosis in hepatocytes that is dependent on p53. AB - Prolonged activation of the mitogen-activated protein kinase (MAPK) pathway enhances expression of the cyclin kinase inhibitor p21 that can promote growth arrest and cell survival in response to cytotoxic insults. Bile acids can also cause prolonged MAPK activation that is cytoprotective against bile acid-induced cell death. Here, we examined the impact of bile acid-induced MAPK signaling and p21 expression on the survival of primary mouse hepatocytes. Deoxycholic acid (DCA) caused prolonged activation of the MAPK pathway that weakly enhanced p21 protein expression. When DCA-induced MAPK activation was blocked using MEK1/2 inhibitors, both hepatocyte viability and expression of p21 were reduced. Surprisingly, constitutive overexpression of p21 in p21+/+ hepatocytes enhanced DCA-induced cell killing. In agreement with these findings, treatment of p21-/- hepatocytes with DCA and MEK1/2 inhibitors also caused less apoptosis than observed in wild-type p21+/+ cells. Expression of p21 in p21-/- hepatocytes did not modify basal levels of apoptosis but restored the apoptotic response of p21-/ cells to those of p21+/+ cells overexpressing p21. These findings suggest that basal expression of p21 plays a facilitating, proapoptotic role in DCA-induced apoptosis. Overexpression of p21 enhanced p53 protein levels. In agreement with a role for p53 in the enhanced apoptotic response, overexpression of p21 did not potentiate apoptosis in p53-/- hepatocytes but, instead, attenuated the death response in these cells. In conclusion, our data suggest that overexpression of p21 can promote apoptosis, leading to elevated sensitivity to proapoptotic stimuli. PMID- 12085348 TI - Ursodeoxycholic acid diminishes Fas-ligand-induced apoptosis in mouse hepatocytes. AB - Ursodeoxycholic acid (UDCA) can protect hepatocytes from apoptosis induced by a variety of stimuli including anti-Fas antibody. However, in vivo the Fas receptor is activated by its natural ligand, Fas-L, whereas anti-Fas antibodies are not a physiologic stimulus. We therefore have assessed the anti-apoptotic effects of UDCA and other bile acids in a novel coculture model where apoptosis is induced in murine hepatocytes by membrane-bound Fas-L expressing fibroblasts. Primary hepatocytes were cultured overnight on collagen-coated coverslips with increasing concentrations of different bile acids and overlaid with either NIH 3T3 Fas-L(+) or Fas-L(-) expressing fibroblasts. After 6 hours cells were fixed and apoptosis was evaluated by TUNEL assay and DAPI staining using digital imaging. Fas-L protein expression and Fas trimerization were evaluated by Western blot analysis. FITC-UDCA and Mitotracker Red were used to evaluate UDCA localization with mitochondria. UDCA (up to 100 micromol/L, P <.0001), TUDCA (up to 400 micromol/L, P <.0001), and TCDCA (up to 200 micromol/L, P <.0001), but not TCA (up to 500 micromol/L), significantly protected hepatocytes in Fas-L(+) cocultures. UDCA had no significant effect on hepatocytes in Fas-L(-) cocultures. TUDCA, 50 micromol/L (P <.001) and TCDCA up to 200 micromol/L (P <.0001) also reduced the hepatocytes apoptotic rate in Fas-L(-) cocultures. Bile acids did not affect Fas-L expression in fibroblasts or Fas trimerization. FITC-UDCA colocalized with the mitochondrial probe. In conclusion, UDCA, TUDCA, and TCDCA but not TCA are capable of protecting hepatocytes from Fas-L-induced apoptosis. This protective effect is not associated with reductions in Fas trimerization, but may be related to a direct effect on the mitochondrial membrane. PMID- 12085349 TI - Reduced glutathione depletion causes necrosis and sensitization to tumor necrosis factor-alpha-induced apoptosis in cultured mouse hepatocytes. AB - The effect of reduced glutathione (GSH) depletion by acetaminophen (APAP), diethylmaleate (DEM), or phorone on the mode of cell death and susceptibility to tumor necrosis factor (TNF)-induced cell death was studied in cultured mouse hepatocytes. Dose-dependent necrosis was the exclusive mode of cell death with APAP alone, but the addition of TNF-alpha induced a switch to about half apoptosis without changing total loss of viability. This effect was seen at 1 and 5 mmol/L but was inhibited at 10 and 20 mmol/L APAP. The switch to apoptosis was associated with increased caspase activities, release of cytochrome c, and DNA laddering and was inhibited by caspase inhibitors. DEM and phorone also induced dose-dependent necrosis. Treatment with TNF-alpha under these conditions lead to incremental cell death in the form of apoptosis at 0.25 and 0.5 mmol/L DEM and 0.1 and 0.2 mmol/L phorone. At 1.0 and 2.0 mmol/L DEM and 0.5 mmol/L phorone, 90% to 100% necrosis was observed with resistance to TNF-alpha effects. The apoptosis with TNF-alpha plus DEM was confirmed by DNA laddering and inhibition by caspase inhibitors. However, in the presence of caspase inhibitors, the increment in cell death induced by TNF-alpha persisted as an increase in necrosis. A combination of antioxidants, vitamin E, and butylated hydroxytoluene (BHT) markedly inhibited necrosis induced by APAP or DEM alone, but the sensitization to TNF-alpha-induced apoptosis was unaffected. GSH monoethylester (GSH-EE) protected against necrosis and apoptosis. In conclusion, depletion of GSH by APAP, DEM, or phorone causes oxidative stress-induced necrosis and sensitizes to an oxidative stress independent TNF-alpha-induced apoptosis. PMID- 12085350 TI - Mrp2 modulates the activity of chloride channels in isolated hepatocytes. AB - Adenosine triphosphate binding cassette family transport proteins are important organic ion transporters in hepatocytes but these molecules may also exhibit other functions. In the present study we have measured the effects of substrates of the canalicular organic ion transporter multidrug resistance associated protein 2 (Mrp2) on chloride channel activation and cell volume regulation. We found that substrates such as leukotriene D(4), 17-beta-estradiol glucuronide, and the leukotriene inhibitor MK-571 accelerated the activation of chloride channels by cell swelling and activated chloride channels in cytokine-pretreated hepatocytes. Two conjugated estrogens that are not Mrp2 substrates did not produce this effect. Hepatocytes derived from a strain of transport-deficient rats (TR(-)), which lack Mrp2 expression, showed none of these substrate effects. Coincident with their ability to activate channels, the Mrp2 substrates increased the rate of volume regulatory decrease by approximately 50% (P <.01), confirming that enhanced channel activation under this condition stimulated volume regulation. In TR-hepatocytes the Mrp2 substrate had no effect on volume regulation. In conclusion, Mrp2 plays a role in regulation of chloride channel function by reducing the lag time necessary for channel activation and consequently accelerating the process of cell volume regulation. Substrates of Mrp2 affect the ability of the protein to interact with chloride channels. These findings represent an alternative function of Mrp2 in hepatocytes. PMID- 12085351 TI - Repeated fasting stress causes activation of mitogen-activated protein kinases (ERK/JNK) in rat liver. AB - Mitogen-activated protein kinases (MAPK)-signaling pathways play key roles in cytoplasmic-nuclear signal transmission in response to various extracellular stimuli. In this study, we investigated the effect of repeated fasting stress on activation of the 3 members of the MAPK family, the extracellular signal regulated kinase (ERK), the c-Jun NH(2)-terminal kinase (JNK), and the p38 mitogen-activated protein kinase (p38 kinase), in rat liver. Immunecomplex kinase assays showed that ERK and JNK were significantly activated in the liver extract from fasted rats whereas p38 kinase showed no activation. In an immunohistochemical study, the phosphorylated and activated form of ERK (p-ERK) was abundantly expressed in pericentral hepatocytes of fasted liver compared with those of the control. On the other hand, the phosphorylated and activated form of JNK (p-JNK) was highly expressed in irregular-shaped cells along the sinusoidal lining of fasted liver. A double immunofluorescent study to identify p-JNK immunoreactive cells revealed them to be Kupffer cells, which are the resident hepatic macrophages. In conclusion, ERK and JNK are selectively activated in distinct cell types of rat liver by repeated fasting stress. PMID- 12085353 TI - Hepatic ischemic preconditioning in mice is associated with activation of NF kappaB, p38 kinase, and cell cycle entry. AB - A brief period of hepatic ischemia protects the liver against subsequent ischemia reperfusion (IR) injury, but the mechanism of such preconditioning is poorly understood. We examined whether preconditioning activated nuclear factor kappa B (NF-kappaB), the stress-activated protein kinases (SAPK), c-Jun N-terminal kinase 1 (JNK-1) and p38, and entry into the cell cycle. We used a murine model of partial hepatic ischemia. Preconditioning was performed by clamping the vasculature for 2 to 20 minutes, and allowing reperfusion for 10 minutes before 90-minute ischemia or IR. As assessed by serum alanine aminotransferase (ALT) levels and liver histology, preconditioning periods of 5 and 10 minutes were highly protective against IR injury, whereas 2-, 15-, and 20-minute intervals were ineffective. Preconditioning was associated with entry of hepatocytes into the cell cycle within 2 hours of subsequent IR, as indicated by proliferating cell nuclear antigen (PCNA) nuclear staining, induction of cyclin D1 and numerous mitotic figures; in the absence of preconditioning, such changes were not seen until 24 hours. Preconditioning increased nuclear binding of NF-kappaB within 30 minutes of the subsequent ischemic interval, paralleled by degradation of inhibitory (binding) protein for NF-kappaB (IkappaBalpha). Ischemic preconditioning also activated p38 kinase and JNK-1, which are known to converge on cyclin D1 regulation. The protective effect of the preconditioning regimen was more closely associated with p38 kinase than JNK-1 activation. In conclusion, the hepatoprotective effects of ischemic preconditioning are associated with activation of NF-kappaB and SAPKs that are associated with entry of hepatocytes into the cell cycle, a critical biological effect that favors survival of the liver against ischemic and IR injury. PMID- 12085352 TI - Peroxisomal localization of inducible nitric oxide synthase in hepatocytes. AB - Shock states induce the expression of inducible nitric oxide synthase (iNOS) in both Kupffer cells and hepatocytes in the liver, but little is known about its subcellular localization in these cells. Studies were undertaken to characterize the subcellular location of iNOS in hepatocytes in response to sepsis. By immunofluorescence analysis, intraperitoneal challenge with bacterial lipopolysaccharide induced cytosolic iNOS in Kupffer cells but punctate labeling in hepatocytes. Cultured rat hepatocytes exposed to interferon gamma, interleukin 1, and tumor necrosis factor alpha showed iNOS protein expression within peroxisomes as early as 4 hours after stimulation, as determined by colabeling for catalase or PMP70. To a lesser extent, iNOS was also observed associated with the plasma membrane and in undefined intracellular aggregates. The nitric oxide synthase (NOS) antagonist L-N-imino-ornithine (L-NIO) did not affect the expression of iNOS within peroxisomes, cytoplasmic aggregates, or cytosol but increased plasma membrane localization of iNOS. Human iNOS transduced into iNOS null mouse hepatocytes using an adenoviral vector also localized to peroxisomes. The expression of iNOS often resulted in the disappearance of detectable catalase in many hepatocytes. In conclusion, these studies establish the peroxisome as a site of iNOS localization in hepatocytes and show a relationship between iNOS up regulation and decreased expression of catalase. PMID- 12085354 TI - Transferrin receptor gene expression and transferrin-bound iron uptake are increased during postischemic rat liver reperfusion. AB - Iron-catalyzed production of reactive oxygen species is a cause of liver injury after ischemia/reperfusion (I/R). The aim of the present study was to address the regulation of transferrin receptor (TfR), which mediates cellular iron uptake, during I/R. The molecular mechanisms controlling TfR gene expression in vivo during I/R of rat liver were investigated by molecular biology procedures. We also analyzed transferrin-bound iron uptake into surviving liver slices. Increased amounts of TfR protein and messenger RNA (mRNA) were found 2 to 6 hours after reestablishment of blood supply. RNA bandshift analysis showed that iron regulatory protein (IRP) activity was decreased in the first hours of reperfusion, thus indicating that IRP-mediated mRNA stabilization was not involved in early TfR upregulation. On the contrary, increased transcription of the TfR gene in isolated nuclei was observed during reperfusion; during the ischemic phase this was preceded by enhanced binding of hypoxia inducible factor (HIF-1) to a DNA sequence derived from the TfR promoter. TfR2 mRNA levels were also enhanced after reperfusion. The increased expression of TfR at the cell surface resulted in increased uptake of transferrin-bound-iron into surviving liver slices; however, iron was not incorporated into ferritin. In conclusion, HIF-1 mediated activation of TfR gene transcription and IRP-mediated increase of TfR mRNA stability ensure a steady induction of TfR, and hence higher iron uptake in reperfused rat liver. TfR-mediated entry of the metal into liver cells may represent a source of catalytically active iron, which may play a role in reperfusion damage. PMID- 12085355 TI - Cyclophosphamide disrupts hepatic sinusoidal endothelium and improves transplanted cell engraftment in rat liver. AB - To determine whether disruption of the hepatic sinusoidal endothelium will facilitate engraftment of transplanted cells, we treated Fischer 344 (F344) rats lacking dipeptidyl peptidase IV (DPPIV) activity with cyclophosphamide (CP). Electron microscopy showed endothelial injury within 6 hours following CP, and, after 24 and 48 hours, the endothelium was disrupted in most hepatic sinusoids. CP did not affect Kupffer cell function. Similarly, CP had no obvious effects on hepatocytes. Intrasplenic transplantation of F344 rat hepatocytes followed by their localization with DPPIV histochemistry showed 3- to 5-fold increases in the number of transplanted cells in CP-treated animals. Transplanted cells integrated in the liver parenchyma more rapidly in CP-treated animals, and hybrid bile canaliculi developed even 1 day after cell transplantation, which was not observed in control animals. To demonstrate whether improved cell engraftment translated into superior liver repopulation, recipient animals were conditioned with retrorsine and two-thirds partial hepatectomy (PH), which induces transplanted cell proliferation. CP treatment of these animals before cell transplantation significantly increased the number and size of transplanted cell foci. In conclusion, disruption of the hepatic sinusoidal endothelium was associated with accelerated entry and integration of transplanted cells in the liver parenchyma. These results provide insights into hepatocyte engraftment in the liver and will help in optimizing liver-directed cell therapy. PMID- 12085356 TI - Production of a cytochrome P450 2E1 transgenic mouse and initial evaluation of alcoholic liver damage. AB - Hepatic metabolism of ethanol by cytochrome P450 2E1 (CYP2E1) is believed to contribute to alcoholic liver damage. To further evaluate CYP2E1 in alcoholic liver disease, we created a transgenic mouse containing human CYP2E1 complementary DNA (cDNA) under the control of mouse albumin enhancer-promoter. Two experiments were performed. In the first experiment, transgenic and nontransgenic mice were fed normal chow. In the second experiment, transgenic and nontransgenic mice were pair fed a nutritionally complete liquid diet for 16 weeks. The liquid diet contained 30% of calories as ethanol (or dextrose) and 25% of calories as corn oil. Liver damage was assessed by measuring serum alanine aminotransferase (ALT) levels and examining liver histology. Transgenic animals reproduced and were phenotypically normal. Hepatic levels of CYP2E1 messenger RNA (mRNA), protein, and enzyme activity did not differ between chow-fed transgenic and nontransgenic mice. Livers from transgenic mice fed the alcohol diet contained significantly more CYP2E1 protein and enzyme activity than livers from nontransgenic mice fed the same diet. Transgenic mice receiving the alcohol diet had significantly higher serum ALT levels than nontransgenic mice. Histologic examination of the livers showed higher histologic scores in transgenic mice fed ethanol compared with nontransgenic mice fed ethanol. Ballooning hepatocytes were seen in livers from transgenic mice fed ethanol. Apoptosis, as determined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) assay, did not differ between groups. In conclusion, we have produced a transgenic mouse that expresses human CYP2E1 in the liver. When fed a nutritionally complete alcohol diet, transgenic mice develop more liver damage than nontransgenic mice. PMID- 12085357 TI - Detection and identification of bacterial DNA in patients with cirrhosis and culture-negative, nonneutrocytic ascites. AB - The current pathogenic theory of spontaneous bacterial peritonitis (SBP) in patients with cirrhosis and ascites suggests that repeated episodes of bacterial translocation (BT) from intestinal lumen to mesenteric lymph nodes followed by systemic seeding are the key steps for the final development of infectious events. However, most of the episodes of systemic bacterial circulation remain undetected. Therefore, we investigated the hypothetical presence of bacteria in blood and/or ascitic fluid (AF) from patients with cirrhosis and sterile (culture negative) AF by means of bacterial DNA (bactDNA) detection and identification. Twenty-eight consecutively admitted patients with cirrhosis and presence of AF were included in the study. BactDNA was detected using a polymerase chain reaction (PCR)-based method. The corresponding bacteria were identified by nucleotide sequencing of purified PCR products. BactDNA was detected simultaneously in blood and AF in 9 patients (32.1%). DNA sequencing allowed the identification of Escherichia coli (n = 7) and Staphylococcus aureus (n = 2). In all cases, the similarity between the sequence found in AF and blood indicated that the bactDNA present in both locations originated from a single clone (single translocation event). Child-Pugh score and basic hemodynamic, clinical, endoscopic, and biochemical characteristics were similar among patients with or without the presence of bactDNA. In conclusion, we have detected bactDNA in serum and AF in 32% of all patients studied, and this likely represents single clone episodes of translocation and systemic seeding. E. coli is the most frequently identified bacteria. PMID- 12085359 TI - Is obesity an independent risk factor for hepatocellular carcinoma in cirrhosis? AB - Recently, several epidemiologic observations have suggested that obesity might be an independent risk factor for certain malignancies such as breast cancer, colon cancer, renal cell carcinoma, and esophageal adenocarcinoma. However, there are no studies examining the risk of hepatocellular carcinoma (HCC) in obesity. The aim of the present study was to determine whether obesity is an independent risk factor for HCC in patients with cirrhosis. Explanted liver specimens from a national database on patients undergoing liver transplantation were examined for HCC, and the incidence was compared among patients with varying body mass indices according to the etiology of cirrhosis. A multivariate analysis was used for controlling other potentially confounding variables such as age and sex. Among 19,271 evaluable patients, the overall incidence of HCC was 3.4% (n = 659) with a slightly higher prevalence among obese patients compared with lean patients. Obesity was an independent predictor for HCC in patients with alcoholic cirrhosis (odds ratio [OR], 3.2; 95% CI, 1.5-6.6; P =.002) and cryptogenic cirrhosis (OR, 11.1; 95% CI, 1.5-87.4; P =.02). Obesity was not an independent predictor in patients with hepatitis C, hepatitis B, primary biliary cirrhosis, and autoimmune hepatitis. The higher risk of HCC in obese patients is confined to alcoholic liver disease and cryptogenic cirrhosis. In conclusion, more frequent surveillance for HCC may be warranted in obese patients with alcoholic and cryptogenic cirrhosis. However, as this study is based on patients with advanced cirrhosis, our findings need to be confirmed in a broader population of individuals with cirrhosis. PMID- 12085358 TI - HFE mutations, hepatic iron, and fibrosis: ethnic-specific association of NASH with C282Y but not with fibrotic severity. AB - There is conflicting evidence regarding inheritance of hemochromatosis gene (HFE) mutations and influence of hepatic iron deposition as cofactors for development of fibrosis in patients with nonalcoholic steatohepatitis (NASH). We studied hepatic iron content (Perls' stain grade), frequency of HFE mutations, and serum iron indices in 93 patients with NASH from a multiethnic background; 59 (63%) were of Anglo-Celtic origin. Data on C282Y mutations were available for all 93 patients and on H63D for 69 patients. Respective controls were 206 (for C282Y, 141 [69%] of whom were Anglo-Celtic) and 180 (for H63D) blood donors. Hyperferritinemia was present in 38 patients (40%) with NASH, but transferrin saturation was increased (>55%) in only 5 (5%). Liver biopsy specimens showed advanced fibrosis in 31 (33%) (cirrhosis in 20%). Altogether, 9 biopsy specimens (10%) showed increased iron: 7 (8%) with grade 2 and 2 (2%) with grade 3 iron staining. Only 1 biopsy specimen with increased iron showed advanced fibrosis. The frequency of C282Y heterozygosity was increased in Anglo-Celtic patients with NASH compared with ethnic blood donor controls (22% vs. 9.2%; P =.035); there were no C282Y homozygotes in the NASH cohort. Although there was a trend toward higher serum ferritin levels among C282Y heterozygotes with NASH, there were no differences in histologic grades of steatosis, inflammation, or fibrosis between individuals with and without C282Y. The frequencies of compound C282Y/H63D heterozygotes (n = 1) or H63D heterozygotes (n = 10) were not increased in NASH. Multivariate analysis identified female sex, diabetes mellitus, and more severe liver inflammation but not HFE mutations, serum ferritin, iron saturation, or hepatic iron staining as independent predictors of hepatic fibrosis. In conclusion, hepatic iron is not a factor linked to hepatic fibrogenesis in patients with NASH. HFE mutations do not confer an additional risk of hepatic fibrosis in this disorder. PMID- 12085360 TI - Androgen receptor exon 1 CAG repeat length and risk of hepatocellular carcinoma in women. AB - The androgen receptor (AR) gene is localized on chromosome X, and shorter CAG repeats in exon 1 of the AR gene were recently suggested to increase hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) risk among men. To examine whether the relationship between the AR-CAG repeats and HCC was also evident among women, we conducted a case-control study in Taiwan. The number of AR-CAG repeats was determined for 238 women with HCC and 354 unrelated control subjects (comprising 188 first-degree and 166 nonbiological relatives) selected from female relatives of patients with HCC. Women harboring 2 AR alleles with more than 23 CAG repeats had an increased risk of HCC (age-adjusted odds ratio [OR], 1.82; 95% CI, 1.06-3.14), compared with women with only short alleles or a single long allele. The association between harboring 2 AR alleles containing longer CAG repeats and HCC was more striking among HBV carriers (age-adjusted OR for more than 22 repeats, 2.23; 95% CI, 1.14-4.34) and particularly prominent among HBV carriers under age 53 years (age-adjusted OR, 3.16; 95% CI, 1.13-8.82). When CAG repeats were analyzed as a continuous variable, the increase in HCC risk associated with each incremental repeat in the shorter of 2 alleles in a given genotype was statistically significant among women with a first-degree relative with HCC (age-adjusted OR, 1.18; 95% CI, 1.01-1.37). No such relationship was detected among women without the family history. In conclusion, our observations suggest that the AR-CAG alleles may contribute to HCC predisposition among women through a mechanism different from that for men. PMID- 12085361 TI - Interactions of rifamycin SV and rifampicin with organic anion uptake systems of human liver. AB - The antibiotics rifamycin SV and rifampicin substantially reduce sulfobromophthalein (BSP) elimination in humans. In rats, rifamycin SV and rifampicin were shown to interfere with hepatic organic anion uptake by inhibition of the organic anion transporting polypeptides Oatp1 and Oatp2. Therefore, we investigated the effects of rifamycin SV and rifampicin on the OATPs of human liver and determined whether rifampicin is a substrate of 1 or several of these carriers. In complementary RNA (cRNA)-injected Xenopus laevis oocytes, rifamycin SV (10 micromol/L) cis-inhibited human organic anion transporting polypeptide C (SLC21A6) (OATP-C), human organic anion transporting polypeptide 8 (SLC21A8) (OATP8), human organic anion transporting polypeptide B (SLC21A9) (OATP-B), and human organic anion transporting polypeptide A (SLC21A3) (OATP-A) mediated BSP uptake by 69%, 79%, 89%, and 57%, respectively, as compared with uptake into control oocytes. In the presence of 100 micromol/L rifamycin SV, BSP uptake was almost completely abolished. Approximate K(i) values were 2 micromol/L for OATP-C, 3 micromol/L for OATP8, 3 micromol/L for OATP-B and 11 micromol/L for OATP-A. Rifampicin (10 micromol/L) inhibited OATP8-mediated BSP uptake by 50%, whereas inhibition of OATP-C-, OATP-B-, and OATP-A-mediated BSP transport was below 15%. 100 micromol/L rifampicin inhibited OATP-C- and OATP8-, OATP-B- and OATP-A-mediated BSP uptake by 66%, 96%, 25%, and 49%, respectively. The corresponding K(i) values were 17 micromol/L for OATP-C, 5 micromol/L for OATP8, and 51 micromol/L for OATP-A. Direct transport of rifampicin could be shown for OATP-C (apparent K(m) value 13 micromol/L) and OATP8 (2.3 micromol/L). In conclusion, these results show that rifamycin SV and rifampicin interact with OATP-mediated substrate transport to different extents. Inhibition of human liver OATPs can explain the previously observed effects of rifamycin SV and rifampicin on hepatic organic anion elimination. PMID- 12085362 TI - Recipient-derived hepatocytes in liver transplants: a rare event in sex mismatched transplants. AB - Bone marrow-derived stem cells have been shown to engraft and populate native tissues during repair and in transplanted animal tissues. Very few studies have been performed in humans to evaluate the possibility of stem cell engraftment in transplanted tissues. In human renal transplants, recipient cells have been demonstrated within vascular and interstitial structures. In a previous study of patients with hepatic transplants, hepatocytes with XY chromosome patterns have been detected in sex-mismatched female to male transplanted livers in a small number of cases. Because of the possibility of Y chromosome microchimerism of females with male offspring, we analyzed the presence of X and Y chromosomes in liver biopsies of 13 patients with sex-mismatched liver transplants (8 female to male, 5 male to female) and long transplant to biopsy intervals (1.2 to 12 years; mean, 4.5 years). We were able to detect recipient-specific sex chromosomal patterns in inflammatory cells by fluorescent in situ hybridization/immunohistochemistry combination within the liver parenchyma but not within hepatocytes. In conclusion, recipient engraftment of stem cells may be an early feature in liver transplant but may be an infrequent persistent feature in long-term grafts. PMID- 12085363 TI - Management strategies using pharmacogenomics in patients with severe HCV-1b infection: a decision analysis. AB - The management of interferon (IFN) therapy for histologically severe chronic hepatitis C virus genotype 1b (HCV-1b [F3]) is controversial. A decision analysis using the Markov decision analysis model was performed for 6 disease management strategies by using clinical data from a Japanese teaching hospital and available published data. The results of base case analyses showed that IFN monotherapy was considered favorable for patients aged 40 to 60 years with HCV-1b (F3). For the sensitivity analyses, to support the results of base case analyses, HCV-1b (F3) patient quality-of-life (QOL) score must be 0.5 or greater for those 40 to 50 years old and 0.4 to 0.5 or greater for those 60 years old. When patients with HCV-1b (F3) were judged as nonresponsive (NR) after IFN monotherapy, the transition probabilities of liver diseases at 40, 50, and 60 years of age had to be such that the progression of liver diseases was controlled at an annual rate of 7.51% to 8.82% or lower, 7.77% to 8.27% or lower, and 6.39% to 6.60% or lower, respectively, and the sustained virologic response (SVR) rate for IFN monotherapy must be 3.0% to 5.51% or greater, 5.57% to 5.93% or greater, and 10.6% to 11.21% or greater, respectively. It is likely that IFN monotherapy could be applied to patients with HCV-1b (F3) aged 40 years at a dose of at least 432 MU. However, IFN monotherapy did not appear useful for patients with HCV-1b (F3) aged 50 and 60 years if they had no amino acid mutation in NS5A 2209 to 2248 and HCV RNA levels exceeded 1.0 mEq/mL. In conclusion, use of decision analysis models can help in therapeutic decisions for patients with HCV-1b. PMID- 12085364 TI - Predictors of HBeAg loss after lamivudine treatment for chronic hepatitis B. AB - Elevated alanine transaminase (ALT) levels and low serum hepatitis B virus (HBV) DNA predict a higher likelihood of hepatitis B e antigen (HBeAg) loss in patients with chronic hepatitis B treated with interferon. Predictors of HBeAg loss in patients treated with lamivudine are not known. The objective of this analysis of 4 lamivudine-controlled Phase III trials was to determine patient-dependent or laboratory variables that predict HBeAg loss. Predictors of HBeAg loss in patients treated with interferon, lamivudine plus interferon, or placebo are also described. A total of 805 adults with chronic hepatitis B were treated either with lamivudine (n = 406), matching placebo (n = 196), interferon (n = 68), or the combination of lamivudine plus interferon (n = 135). Demographic and baseline disease characteristics were used in stepwise multivariate analyses to identify features that were predictive of lamivudine-induced HBeAg loss. HBeAg loss correlated with increased pretreatment ALT levels in all groups. The rate of HBeAg loss was highest among patients with pretreatment ALT levels greater than 5 times the upper limit of normal (ULN) and was most pronounced in the lamivudine group (56%). Multivariate modeling indicated that elevated baseline ALT levels (P <.001) and histologic activity index (HAI) score (P <.001) were important predictors of HBeAg loss in response to lamivudine. The effect of pretreatment ALT levels on HBeAg loss was similar for Asians and Caucasians. In conclusion, elevated pretreatment ALT levels and/or active histologic disease were the most important predictors of lamivudine-induced HBeAg loss. Asians and Caucasians had similar rates of response to lamivudine at comparable ALT levels. PMID- 12085365 TI - Polymorphisms of microsomal epoxide hydrolase gene and severity of HCV-related liver disease. AB - Factors influencing the progression of liver disease and the development of hepatocellular carcinoma (HCC) in chronic hepatitis C virus (HCV) infection are poorly understood. Inherited variations of drug-metabolizing enzyme (DME) activities may affect liver damage and cancer risk by modifying individual susceptibility to endogenous or exogenous toxic compounds. We investigated the association of liver disease severity with common alleles of microsomal epoxide hydrolase (mEH), an enzyme involved in the metabolism of highly reactive epoxide intermediates. Three polymorphisms (Tyr113His, His139Arg, and -613C/T) were analyzed by polymerase chain reaction (PCR) restriction fragment length polymorphisms (RFLPs) in 394 patients at different stages of disease, including 92 asymptomatic carriers, 109 patients with chronic hepatitis, 100 patients with cirrhosis, and 93 patients with HCC. Reference allele frequencies were obtained from 99 healthy blood donors. Allele distributions between categories were compared using the chi(2) test; odds ratios (ORs) and 95% CI were calculated to express relative risks. Allele frequencies among 99 healthy controls were as follows: 15.1% for 113His/His, 4.0% for 139Arg/Arg, and 46.5% for -613C/T. mEH 113His/His homozygotes were overrepresented in advanced stages of disease, in particular among HCC patients (27.9%; P =.03; OR, 2.2; 95% CI, 1.0-4.6). Differences were more pronounced among men and between extreme patient categories. When mEH genotypes were combined to express a metabolic phenotype, very slow metabolizers were highly prevalent among cirrhotic and HCC patients (18% vs. 3.3% in carriers; P <.001). In conclusion, mEH gene polymorphisms were significantly associated with HCV-related liver disease severity and HCC risk. Men were at higher risk than women; this might be explained by hormonal regulation of gene expression or by differential exposure to environmental toxins. PMID- 12085366 TI - Contribution of donor age to the recent decrease in patient survival among HCV infected liver transplant recipients. AB - Recurrent hepatitis occurs in the majority of patients undergoing liver transplantation for hepatitis C virus (HCV) cirrhosis, with progression to cirrhosis in up to 30% after 5 years. Based on these data, a decrease in survival can be anticipated with prolonged follow-up. Furthermore, posttransplantation HCV fibrosis progression has been shown in recent years to increase. Our aims were (1) to describe the natural history of HCV-infected recipients, particularly to determine whether survival has decreased in recent years; (2) to compare this outcome with that observed in non-HCV-infected cirrhosis controls; and (3) to determine the factors associated with disease severity and survival. Among 522 cirrhotic patients undergoing transplantation between 1991 and 2000, 283 (54%) were infected with HCV. Yearly biopsies were performed in these recipients and at 1 and 5 years in the remainder. With similar follow-up, the percentage of deaths in the HCV(+) group was significantly higher than in the HCV- group (37% vs. 22%, P <.001), and patient survival was lower (77%, 61%, 55% vs. 87%, 76%, 70% at 1, 5, and 7 years, respectively; P =.0001). Although survival has increased in the HCV- group in recent years, it has significantly decreased in HCV recipients (P <.0001). The main cause of death among the latter was decompensated graft cirrhosis (n = 23/105, 22%), whereas that of HCV- patients was infections (n = 10/52, 19%). Reasons for the recent worse outcome in HCV+ recipients include the increased donor age and stronger immunosuppression. In conclusion, patient survival is lower among HCV+ recipients than among HCV- ones and has been decreasing in recent years. The aging of donors is a major contributor to this worse outcome. PMID- 12085367 TI - Clinical utility of total HCV core antigen quantification: a new indirect marker of HCV replication. AB - Hepatitis C virus (HCV) RNA detection, viral load quantification, and HCV genotyping are widely used in clinical practice. Recently, the availability of an anticore antigen (Ag) monoclonal antibody allowed development of an enzyme-linked immunosorbent assay (ELISA) detecting and quantifying total HCV core Ag in peripheral blood of HCV-infected patients. The aims of the present study were to investigate the biologic significance of this new marker in HCV infection, to establish the intrinsic performance of the current assay, and to determine its potential utility in the management of HCV-infected patients. A panel of infected sera calibrated to the World Health Organization International Standard and 657 serum samples from infected patients receiving antiviral treatment were studied. We showed that total HCV core Ag quantification is an accurate, precise, and specific indirect marker of HCV replication. We estimated that 1 pg/mL of total HCV core Ag is equivalent to approximately 8,000 HCV RNA international units (IU)/mL, although minor between-patient differences may exist. In conclusion, total HCV core Ag quantification can be used in the various indications of viral load monitoring, including the evaluation of baseline viral load before therapy, the assessment of the virologic response to antiviral treatment, and the study of early viral kinetics during therapy. Nevertheless, the total HCV core Ag assay cannot be used as a marker of viral replication for HCV RNA values below 20,000 IU/mL, limiting its use in the monitoring of late events during and after antiviral treatment. PMID- 12085368 TI - Course of virologic breakthroughs under long-term lamivudine in HBeAg-negative precore mutant HBV liver disease. AB - We studied the course of virologic breakthroughs detected by a quantitative polymerase chain reaction (PCR) assay in 32 of 78 patients with hepatitis B e antigen (HBeAg)-negative precore mutant hepatitis B virus (HBV) chronic liver disease under long-term lamivudine monotherapy. Serum HBV DNA levels were measured every 3 months and on every biochemical breakthrough. YMDD mutants were detected in 30 of the 32 patients with virologic breakthroughs. Among these 32 patients, biochemical remission rate was 44% at 6 months, 21% at 12 months, and 0% at 24 months after the onset of virologic breakthrough. Development of biochemical breakthroughs was associated with a significant increase of serum HBV DNA levels, which exceeded 100,000 copies/mL in 19 of 20 patients (95%) with biochemical breakthroughs and in only 1 of 8 patients (12.5%) remaining in biochemical remission for at least 6 months after the onset of virologic breakthrough (P <.001). Alanine aminotransferase (ALT) level peaked within 0 to 3 months after the onset of biochemical breakthrough and decreased at 6 months but remained abnormal in all but 2 patients. Follow-up liver histologic lesions in patients with biochemical breakthroughs did not differ from baseline findings, although they were significantly improved in patients remaining in virologic and biochemical remission. In conclusion, the frequent emergence of viral resistance under long-term lamivudine monotherapy in HBeAg-negative precore mutant HBV chronic liver disease is followed by increasing viremia levels culminating in the development of biochemical breakthroughs in most cases. ALT activity peaks close to the onset of biochemical breakthrough, decreasing thereafter but remaining persistently abnormal with fluctuating levels. PMID- 12085369 TI - Burden of liver disease in the United States: summary of a workshop. PMID- 12085370 TI - Antioxidant gene therapy and hepatic ischemia-reperfusion injury. PMID- 12085371 TI - Leptin, liver, and obese mice--fibrosis in the fat lane. PMID- 12085372 TI - Transplantation into and inside the liver. PMID- 12085373 TI - Pharmacogenomics of hepatitis C and decision analysis: a glimpse into the future. PMID- 12085374 TI - Intentional coinfection of patients with HCV infection using avian infection bursal disease virus. PMID- 12085375 TI - Lack of evidence for ribavirin-induced bone loss. PMID- 12085376 TI - Hepatitis B vaccination after liver transplantation. PMID- 12085378 TI - Is TIPS a cost-effective therapy for the prevention of variceal rebleeding? PMID- 12085380 TI - Out came copious water. PMID- 12085384 TI - Does vascular access monitoring work? AB - It is widely accepted that surveillance with correction of stenosis before thrombosis prolongs hemodialysis graft survival. However, few prospective randomized controlled trials have confirmed the efficacy of this approach. Most studies have been nonrandomized or have included historic rather than concurrent controls. Although graft dysfunction as assessed by blood flow determination has been widely recognized as the surveillance method of choice, all fully randomized prospective studies that have found a benefit in terms of decreased thrombosis or increased survival have evaluated stenosis and the benefit has only been in new grafts. Finally, the accuracy of blood flow measurements as determined by ultrasound dilution in predicting thrombosis is controversial. Some have reported that predictive accuracy is adequate, whereas others have reached the opposite conclusion. Thus, further studies are needed to explore the effectiveness of surveillance with intervention in improving graft survival. Until such studies establish the use of surveillance programs, surveillance of all grafts using blood-flow monitoring cannot be supported. PMID- 12085383 TI - Hemodialysis vascular access dysfunction from basic biology to clinical intervention. AB - Hemodialysis vascular access dysfunction is a major cause of morbidity and hospitalization in the hemodialysis population at a cost of over 1 billion dollars per annum. Venous stenosis and thrombosis as a result of venous neointimal hyperplasia are the major causes of hemodialysis vascular access dysfunction. Despite the magnitude of the clinical problem, there are currently no effective therapies for this condition. We believe that this could be because of an inadequate understanding of the pathogenesis of this condition. At a histological level, venous neointimal hyperplasia (both in human specimens and in a pig model) is characterized by the presence of smooth muscle cells/myofibroblasts, microvessel formation (angiogenesis), and the accumulation of extracellular matrix components, all of which could be potential targets for therapeutic intervention. In particular, polytetrafluoroethylene dialysis access grafts could be the ideal clinical model for testing out novel local therapies to block neointimal hyperplasia. The current review describes the lesion of venous neointimal hyperplasia in human samples and in a pig model and suggests possible future directions for the development of effective local therapies for this condition. PMID- 12085385 TI - Are hemodialysis access flow measurements by ultrasound dilution the standard of care for access surveillance? AB - Maintenance of vascular access function is vital to the delivery of adequate hemodialysis therapy. Failure of function is associated with significant morbidity and cost. Thus, access surveillance programs are suggested. The most common cause for access dysfunction is stenosis formation within the graft fistula. This may lead to reduced blood flow. The measurement of access blood flow has thus been recommended as the preferred method for surveillance. This article reviews blood flow among other methods for the screening of access dysfunction, the techniques used to measure it, the predictability of access flow measurements in determining the presence of access stenosis and allowing successful; intervention and finally the cost-effectiveness of such surveillance. Review of available evidence would suggest that access flow measurements are the best tests currently available to screen for access dysfunction, and as preventative interventions, such as angioplasty and surgery, are successful, they should be regarded as the present standard of care. This would appear to be a cost-effective strategy. Furthermore, the method of choice for access flow measurement is by ultrasound dilution technology. PMID- 12085386 TI - Endovascular intervention for the failing vascular access. AB - Endovascular interventions have replaced surgical repair as the primary treatment of the failing or thrombosed vascular access. However, endovascular and surgical techniques are complementary: the limitations of one are the strengths of the other. Endovascular management of access-related problems is a critical component of a successful vascular access monitoring program. The identification and early treatment of developing stenoses, before access thrombosis are essential roles that are ideally suited to percutaneous, image-guided techniques. Despite recent advances in endovascular techniques and devices, angioplasty continues to be the primary method for treatment of access-related stenoses. When appropriate lesions are treated, angioplasty is a fast, easy, and safe procedure that can extend to patency of a hemodialysis graft or fistula. PMID- 12085387 TI - Systemic barriers to improving vascular access outcomes. AB - Vascular access dysfunction is the most frequent cause of hospitalization for end stage renal disease (ESRD) patients. Our system of vascular access care and industry standards developed for historic reasons have resulted in a haphazard approach to access management. The Dialysis Outcome Quality Initiative has provided a road map for improving vascular access management. However, despite widespread acceptance, these recommendations are not routinely followed. This is largely the result of inertia coupled with systemic barriers to improving access outcomes. These barriers include lack of funded pre-ESRD care and preoperative imaging, lack of reimbursement for access monitoring, unavailable surgical and interventional suites, erosion of the real value of the composite rate, bundling of additional new services without rate adjustment, poor accountability of surgeons and hospitals, and a reimbursement system that rewards procedures and, in particular, graft and catheter placement. Currently, Center for Medicare and Medicaid Services is reevaluating the composite rate and its included bundle of services. To provide the best access care with the fewest complications while insuring multidisciplinary involvement and accountability, a realistic appraisal and realignment of incentives must be developed to insure improvement of access care in the United States. PMID- 12085388 TI - Moncrief-Popovich catheter and implantation technique: the AV fistula of peritoneal dialysis. AB - Despite the decrease in peritonitis rate from touch contamination caused by the use of disconnect systems, technique failure in peritoneal dialysis (PD) from pericatheter and exit-site infections in PD remains unchanged. This indicates a failure of current PD catheters to prevent bacterial transfer from exit site to the peritoneal cavity. In 1991, Moncrief and Popovich introduced a new catheter design and implantation technique to address this problem. The catheter is made of silastic, has a coiled tip with 2 cuffs, and an arcuate bend between the cuffs. This would prevent catheter malfunction and leakage of PD fluid. The implantation technique involves embedding of the external segment of the catheter in the subcutaneous tunnel at insertion. The catheter segment is kept embedded for 4 to 6 weeks before externalization. This procedure will allow time for tissue ingrowth on into the external cuff and catheter surfaces between the 2 cuffs, preventing bacterial colonization of the catheter surfaces from the exit wound and thereby reducing pericatheter infections. Thus, the new technique will establish a more effective bacteriologic barrier between the exit wound and the peritoneal cavity than the conventional catheters. Ten years after validation of the catheter design and implantation technique by Moncrief and Popovich, various clinical studies confirm that this new technique of catheter implantation increases catheter life expectancy and reduces pericatheter infections in PD. Like the arteriovenous fistula of haemodialysis, this new catheter remains embedded in subcutaneous tunnel, is exteriorized electively when patient needs to be started on dialysis, and reduces pericatheter and exit-site infections in PD. The new technique, therefore, is widely accepted as a simple, safe, and cost effective procedure for quality care of PD patients around the world. PMID- 12085389 TI - Presternal peritoneal catheter. AB - The swan neck presternal catheter is composed of 2 flexible (silicon rubber) tubes joined by a titanium connector at the time of implantation. The exit site is located in the parasternal area. The catheter located on the chest was designed to reduce the incidence of exit site infections compared to peritoneal dialysis catheters with abdominal exits. From August 1991 to September 30, 2001, 974 swan neck presternal catheters were implanted worldwide. At the university of Missouri, 150 of these catheters were implanted and followed for over 130 patient years. Presternal catheters tended to perform better than swan neck abdominal catheters regarding exit and tunnel infections, even though they were implanted in several patients in whom regular catheters with the exit on the abdomen would be difficult or impossible to implant. Two-year survival probability of presternal catheters was 0.95. Recurrent/refractory peritonitis was the only reason for catheter failure. The catheter is particularly useful in obese patients (body mass index >35), patients with ostomies, children with diapers and fecal incontinence, and patients who want to take baths without the risk of exit contamination. Many patients prefer presternal catheter because of better body image. Disadvantages of the presternal catheter are minimal. Compared with abdominal catheters, dialysis-solution flow is slightly slower because of the increased catheter length; however, slower flow is insignificant clinically. There is a possibility of catheter disconnection in the tunnel, but this complication is extremely rare in adults and easily corrected. Finally, the implantation technique is more challenging compared with that of single-piece, abdominal catheters. PMID- 12085390 TI - Clinical trials of the T-fluted (Ash Advantage) peritoneal dialysis catheter. AB - The Ash Advantage is a T-shaped peritoneal catheter with a single transabdominal tube joining to a tube lying against the parietal peritoneum. Segments with long flutes (grooves) serve as fluid ports rather than 1-mm diameter holes. The folded catheter is placed through the expandable Y-TEC Quill positioned peritoneoscopically, similar to the procedure for a conventional Tenckhoff catheter. In a clinical study, we placed 18 Ash Advantage ("Advantage") catheters, 8 in patients with prior hydraulic or infectious complications of Tenckhoff catheters and 10 in patients initiating peritoneal dialysis. Mean follow-up has been 8.4 months. Outflow rate for the Advantage catheters is uniformly higher than for Tenckhoff catheters at 220 +/- 100 mL/min for the first 5 minutes and 145 +/- 72 mL/min for the next 5 minutes, with outflow completed in 6 to 15 minutes. Variation of outflow volumes for similar exchanges in the same patient is +/- 3% to 5%, much less than with Tenckhoff catheters. Four catheters failed after placement, each for a different reason including outflow failure because of preexisting intraperitoneal adhesions, preexisting peritonitis failing to clear, tubing kink during placement, and catheter break after a freak accident. Life-table analysis of the production version of the catheter in study patients and 12 additional unselected patients indicates a 90% survival at 12 months. In long-term use, the fixed position of the cuffs of this catheter may avoid catheter extrusion, pericatheter hernias, and pericatheter leaks. Multiple fluted ports appear to diminish omental attachment. The catheter may be a useful alternative to conventional Tenckhoff catheters in patients at high risk of catheter failure or in standard peritoneal dialysis patients. PMID- 12085391 TI - Access-related complications in peritoneal dialysis in developing countries. AB - Access-related complications differ considerably between acute and chronic peritoneal dialysis as a result of differences in catheter design, catheter material, number of cuffs, break in period, and the duration of peritoneal dialysis. Pain, which is the commonest complication of acute peritoneal dialysis, is seen in 56% to 75% followed by blood tinged dialysate effluent in 30%, pericatheter leak in 14% to 36%, catheter malfunction in 12% to 28%, bowel injury in 0.1% to 1.3%, rarely bladder injury and peritonitis in 2.5%. In chronic peritoneal dialysis, the incidence of exit-site infections are seen anywhere between 1 episode in 27.3 patients months to 1 episode in 41.9 patients months, and pericatheter leak is seen in 5.2%. Technique of exit-site care is variable in different countries, and regular use of mupirocin ointment at the exit site is practiced in different centers. PMID- 12085392 TI - Incorporating spirituality into the delivery of dialysis care: one team's perspective. AB - The importance of spirituality and organized religion in coping with chronic kidney disease (CKD) and other medical illnesses has been cited in the literature in recent years. This article describes how one dialysis unit recognized the importance of spiritual resources for patients and incorporated the role of chaplain into its interdisciplinary health care team. The resulting enhancement of team functioning and sensitivity to patients' spiritual needs is discussed. PMID- 12085393 TI - The neurological complications of electrical injury: a nursing case management perspective. AB - High-energy electrical injury, whether from lightning strike or electrical shock, occurs primarily in the workplace. Neurological dysfunction can be a devastating complication of electrical injury. A review of the literature was undertaken to develop a better understanding of the epidemiology, mechanisms of injury and neuropathology associated with this type of injury. The numerous challenges inherent in the management of these complex cases were illustrated by three case studies. PMID- 12085394 TI - Pharmacology review: the role of ondansetron in the management of children's nausea and vomiting following posterior fossa neurosurgical procedures. AB - The management of nausea and vomiting is fundamental to the post-operative nursing care of children. Children who have neurosurgical procedures, especially those that involve the posterior fossa, are likely to experience nausea and vomiting in the post-operative period. The proximity of brainstem emetic centres to the surgical site compounds the usual post-operative risk factors for nausea and vomiting. Ondansetron is discussed as an agent that may be more effective than the traditionally-used antiemetics, such as dimenhydrinate and metoclopramide, in this population. Nurses must advocate for effective therapeutic measures to manage children's post-operative nausea and vomiting. Advocacy requires knowledge of high-risk groups, accurate assessment, timely intervention, and thorough evaluation of pharmacological and non-pharmacological measures. PMID- 12085395 TI - Certification ... and other credentials. PMID- 12085396 TI - The perfect storm. PMID- 12085397 TI - Formula for success with safety--just do it! PMID- 12085398 TI - Nursing shortage. PMID- 12085399 TI - Remembering the patient. PMID- 12085400 TI - Preferences in surgical waiting area amenities. AB - The purpose of this needs assessment was to determine which waiting area amenities family members deem important to make them comfortable when waiting for news about surgical patients. A literature review was conducted, perioperative staff members were interviewed and surveyed, and family members were interviewed and surveyed. The findings indicate that family members have preferences for specific amenities. Items of highest importance include the service of a surgical nurse coordinator and a private room for consulting with physicians. Items of average importance include beepers and educational materials about the procedure. Items of least importance include a food cart and soft music. PMID- 12085401 TI - Managing change during an information systems transition. AB - This article examines the social forces underlying computer technology diffusion into nursing. It also explores the benefits of computerized perioperative patient documentation and suggests methods that can used to persuade nurses to embrace and master computer technology as a tool of their trade. Options are examined that offer nurses opportunities to overcome their reluctance to accept responsibility for accurate computer documentation and appreciate the added value offered by information technology. Implementing an adequate computer technical skills training program affects nurses' acceptance of computerized documentation, but having the best training program available does not replace the need to ensure that organizational and individual practitioners are receptive to change. PMID- 12085402 TI - Ethics in perioperative practice--accountability and responsibility. AB - Though often difficult, ethical decision making is necessary when caring for surgical patients. Perioperative nurses have to recognize ethical dilemmas and be prepared to take action based on the ethical code outlined in the American Nurses Association's (ANA's) Code of Ethics for Nurses with Interpretive Statements. In this third of a nine-part series that will help perioperative nurses relate the ANA code to their own area of practice, the author looks at the fourth provision statement, which addresses nurses' accountability and responsibility. PMID- 12085403 TI - Myths and realities of ageism and nursing. AB - Ageism is pervasive throughout society, and it is an especially important issue for nurses because the nursing workforce is aging rapidly. This article explores ageism as it applies to women and nurses, along with the myths and realities about this prevalent attitude. It also presents strategies for managing and educating the aging nursing workforce. PMID- 12085404 TI - The pancreas--hermit of the abdomen. AB - The pancreas is almost impossible to palpate because it is located in the retroperitoneal space. Life-threatening lesions, therefore, often are not diagnosed until they are very advanced and encroaching on adjacent structures. The pancreas has a large endocrine and exocrine reserve, which also contributes to difficulty in diagnosing pancreatic disease. This article provides an in-depth analysis of the anatomy and physiology of the pancreas and endocrinopathies, allowing nurses to provide optimal care for patients with pancreatic abnormalities. PMID- 12085405 TI - Reprocessing single-use devices--the equipment connection. AB - This is the second in a series of three articles about reprocessing medical devices labeled as "single use" by the manufacturer. The goal of reprocessing single-use devices (SUDs) is to save money and decrease environmental pollution. Reprocessing can be performed on SUDs that have been used on other patients or opened but not used. In this article, the procedures for reprocessing (e.g., cleaning, inspecting, sterilizing, tracking, testing, validating) and establishing a reuse program are discussed. The first article of the series, published in the May 2002 issue of the Journal, discussed the ethical component of reusing SUDs. The third article, to be published in the July 2002 issue, will discuss the roles of the involved regulatory agencies and organizations guiding the process. PMID- 12085407 TI - A call to action for perioperative nurses. PMID- 12085406 TI - Evidence-based practice guidelines--one way to enhance clinical practice. AB - Abdominoplasty and liposuction guidelines are just two of the guidelines that can be accessed and used to enhance patient care. Guidelines also can be used to increase your knowledge about many other health care topics. The NGC has approved guidelines for managing chronic pain, as well as guidelines on chronic diseases (e.g., diabetes mellitus, hypertension, chronic obstructive pulmonary disease). Many patients have chronic diseases, and you or your family members also may be affected by chronic disorders. These guidelines provide you with a quick overview of evidence-based treatment protocols. These guidelines are not a panacea for evidence-based practice, but using them is one way that perioperative nurses can enhance their clinical skills. Though not everyone has personal Internet access, most health care facilities do or can make access a reality. Other options include medical or public libraries. Then one simply has to access the NGC web site and join other professionals in improving the quality and timeliness of patient care. PMID- 12085408 TI - Finding patient safety Internet resources. PMID- 12085409 TI - The important role of the Joint Commission. PMID- 12085410 TI - [Health registries, a current necessity]. PMID- 12085411 TI - [We nephrologists who choose the laboratory]. PMID- 12085412 TI - [Do Spanish nephrologists take full advantage of medical informatics?]. PMID- 12085413 TI - [Adequate dialysis]. PMID- 12085414 TI - [TGF-beta: synthesis and mechanism of action]. PMID- 12085415 TI - ["Occult" renal insufficiency due to evaluating renal function using only serum creatinine]. AB - Timely referral to nephrologists depends on identification of renal failure. Most primary care physicians and specialists rely on serum creatinine as the standard test for determination of renal function. Creatinine clearance requires 24 hours urine collection with many pitfalls and wrong results. We compare serum creatinine and the Cockcroft-Gault (C-G) equation as measure of glomerular filtration rate (GFR). The study included 1,053 outpatients with serum creatinine lower than 2.5 mg/dl referred to our nephrological laboratory for serum creatinine and GFR determination using the C-G formula. Patients were grouped into two groups: normal renal function (serum creatinine < 1.3 mg/dl) and "incipient" abnormal renal function (serum creatinine 1.3-2.5 mg/dl). In the group of females with normal creatinine 22% (60-70 y), 35% (70-80 y) and 57% (> 80 y) had GFR values below 50 ml/min. In the group of males 11.3% (70-80 y) and 33.3% (> 80 y) also had GFR reduction in spite of normal serum creatinine. A severe renal insufficiency with creatinine clearance lower than 30 ml/min was observed in the group with "incipient" renal failure based on serum creatinine: 22.7%, 40% and 82.9% for females and 6%, 22.7% and 57% for male (60-70 y; 70-80 y; and > 80 y respectively). In order to improve management and prevention of renal failure appropriate measurements of renal function other than serum creatinine should be emphasize. PMID- 12085416 TI - [Prevalence of renal involvement in a population of type Ii diabetics followed up in primary care]. AB - Patients with type 2 diabetes use to be managed in their primary care settings during the early stages of the disease. The main objective of the study was to determine renal impairment prevalence, and to assess its significance, within type 2 diabetics controlled by their family physicians. PATIENTS AND METHOD: Transverse observation of patients with type 2 diabetes who were the first 20 unselected cases seen by 183 family physicians from 16 of the 17 Autonomic Communities of our country. The following variables were determined: serum creatinine, glucose, and HbA1c concentrations, proteinuria (dipstick test in a first-voided morning urine sample), blood pressure levels, and associated cardiovascular disease. RESULTS: Data from 3,583 type 2 diabetic subjects were evaluated. Mean age was 64 +/- 10 years and 45% were male. A serum creatinine > or = 1.2 mg/dl was observed in 523 (15.5%) patients. Proteinuria was present in 794 (23.5%) cases, being > or = 2 + in 215 (6.5%) subjects. Patients with a serum creatinine > or = 1.2 mg/dl were older, shower higher blood pressure levels, and suffered from more cardiovascular disease (32.0 vs 19.5%) than those with a serum creatinine < 1.2 mg/dl. In a multivariate analysis, this difference continued to be significant (OR 1.47; 95% CI 1.14 to 1.90; p = 0.002. Patients with proteinuria showed a higher prevalence of cardiovascular disease (OR 1.83; 95% CI 1.47 to 2.27; p < 0.0001) than those without proteinuria. This association was continuous through no proteinuria to the > or = 2 + proteinuria (p < 0.001). Blood pressure level was > or = 140/90 mmHg in 69% of the cases, being < 130/85 mmHg in only 8% of the subjects. CONCLUSIONS: There is a high prevalence of renal impairment, approximately of 25% within type 2 diabetic patients seen at the primary care level. Optimal blood pressure level seems to be extremely infrequent bearing in mind the diagnosis of diabetes and the associated cardiovascular disease. PMID- 12085417 TI - [Renal histological lesions in patients with type II diabetes mellitus]. AB - Diabetic glomerulosclerosis is the most frequent cause of renal disease in patients with type II diabetes mellitus (DM), sometimes accompanied by vascular lesions. However, other glomerular pathologies are important in these patients. The aim of this study was to evaluate the prevalence of non-diabetic nephropathy (NDN) in selected patients with type II DM, and to identify clinical markers that may predict its presence in this population. We reviewed 20 renal biopsies performed on twenty patients with type II DM. Nine of them showed diabetic nephropathy (DN) (45%), whereas eleven showed NDN (55%): 1 IgA nephropathy, 3 vasculitis and 7 membranous nephropathy. We found no differences between the two groups with regard to sex, duration of DM, insulin therapy, glycosylated haemoglobin, proteinuria, presence of nephrotic syndrome, hypertension, serum IgA level or renal size. The NDN group had haematuria in 63.6%, whereas the patients with NDN had it in 44.4% (NS). Body mass index was higher in NDN patients (30 +/- 6.7 vs 22 +/- 2.9; p < 0.01), The same was true for creatinine clearance (82.2 +/ 51.4 ml/m vs 40.4 +/- 19.6 ml/m; p < 0.05). The age at the moment of diagnosis was higher in ND patients (67 +/- 11.2 vs 54.3 +/- 4.6; p < 0.05). The 3 patients who had diabetic retinopathy were found to have DN on renal biopsy (diagnostic specificity = 100%), although 66.7% of the patients with diabetic glomerulopathy had no retinopathy. We conclude that patients with type II DM with renal findings suggesting non-diabetic renal disease frequently it have NDN, and a renal biopsy must be performed. The presence of retinopathy has a predictive value of 100% in predicting DN, therefore its existence may make this diagnostic procedure unneccesary. PMID- 12085418 TI - [Effect of antihypertensive combinations on arterial pressure, albuminuria, and glycemic control in patients with type II diabetic nephropathy: a randomized study]. AB - Type II diabetic patients with albuminuria are at high risk for cardiovascular complications; the intense antihypertensive treatment required often involves using drug combinations. The aim of the present study was to compare the effect of two different, renin-angiotensin blocking combinations, on blood pressure (BP), albuminuria and glycemic control. Its design was prospective, randomised, controlled, of parallel branches, and performed in one Endocrinology Department, in Spain. 77 type-II diabetic patients, with stable albuminuria (30-1,000 mg/day) were included. After a pre-inclusion time of 2 weeks, patients were randomised to verapamil SR/trandolapril 180/2 (VT) or losartan/hydrochlorothiazide (LH) 20/12.5 mg/day. Duration of treatment was 1 year. The evaluated parameters were changes in blood pressure, urinary albumin excretion for 24 hours, glycated hemoglobin and plasmatic urea. Overall BP significantly decreased from 161.6 +/- 18.7/83.6 +/- 10.2 mmHg to 137.2 +/- 15.7/70.9 +/- 8.3 mmHg (p < 0.0005). Values, by treatment, were: For VT, 164.3 +/- 18.5/87.2 +/- 10.7 mmHg at baseline and 135.0 +/- 15.1/71.3 +/- 8.4 mmHg at conclusion. For LH, 158.8 +/- 17.4/80.1 +/- 8.4 mmHg at baseline and 139.3 +/- 16.1/70.5 +/- 8.2 mmHg at conclusion. Albuminuria significantly decreased from 308.2 +/- 544.7 mg/day to 198.0 +/- 285.3 mg/day. Both parameters showed no significant difference between treatments. Glycated hemoglobin decreased from 7.59 +/- 1.3% to 7.14 +/- 1.2% in the VT group, and from 7.96 +/- 1.29% to 7.84 +/- 1.62% in the LH group (ANOVA, p = 0.022). Changes adjusted from baseline values showed a trend to the difference between both treatments (p = 0.092). Plasmatic urea increased from 39.8 +/- 12.7 to 40.5 +/- 11.1 mg/dL in the TV group and from 43.4 +/- 12.0 mg/dL to 52.4 +/- 19.4 mg/dL in the LH group (ANOVA, p = 0.028). In conclusion, both treatments reduce blood pressure and albuminuria in a similar way in type II diabetic patients. The verapamil/trandolapril combination contributes to a better carbohydrate metabolism than losartan/hydroclorothiazide. PMID- 12085419 TI - [Comparison of clinical arterial pressure, home-arterial pressure measurement, and ambulatory arterial pressure monitoring in patients with type II diabetes mellitus and diabetic nephropathy]. AB - BACKGROUND: Hypertension is common in type 2 diabetes with diabetic nephropathy, and increases the risk of cardiovascular complications and renal chronic insufficiency. The aim of our evaluation in these patients was: a) to study the correlation between office blood pressure (BP), self-monitored (SMBP) and 24-hour ambulatory blood pressure monitoring (ABPM). b) To study the correlation between these methods and cardiovascular and renal complications. METHODS: We studied 60 patients (mean age 66.7 +/- 9 years, mean duration of diabetes 11.3 +/- 7 years) with arterial hypertension, type 2 diabetes and diabetic nephropathy. Macroangiopathy and echocardiography were recorded. We measured, SMBP and ABPM without modifying the antihypertensive treatment. The white coat phenomenon (WCP) was determined and patients were classified as dippers or non dippers according to their blood pressure diurnal rhythm. RESULTS: Mean glycated haemoglobin was 7.8% and mean serum creatinine 1.2 +/- 0.5 mg/dl, 30% of patients had proteinuria and 70% microalbuminuria The mean number of antihypertensive drugs was 2.2 +/- 1. The mean BP was: Office BP: 158.2 +/- 24/85.3 +/- 9 mmHg, pulse pressure (PP) 72.9 +/- 21 mmHg; SMBP: 145.4 +/- 18/77.5 +/- 7 mmHg, PP 67.9 +/- 18 mmHg and BP in the early morning 150.2 +/- 20/79.9 +/- 9 mmHg; ABPM: diurnal mean 138.9 +/- 15/74.1 +/- 6 mmHg, PP 64.8 +/- 15 mmHg and BP in the early morning 146.5 +/- 16/78.5 +/- 7 mmHg. The three techniques showed a good correlation and WCP was detected in 46.7% of patients with SMBP and in 56.7% with ABPM. We found no correlation between BP and macroangiopathy, but an increase of systolic BP in SMBP and ABPM in proteinuric patients were found and correlation between mass left ventricular index (MLVI) and PP in office and systolic BP and PP in SMBP and ABPM was significant. 70% of patients were non dippers, with a higher MLVI. CONCLUSIONS: Decreases in BP in type 2 diabetes with diabetic nephropathy are difficult of maintain despite combinations of different antihypertensive drugs. These patients present an important WCP and worse prognosis data, such as elevation of systolic BP, increased PP, poor night BP fall and a BP rise in the early morning. Also, we can't reduced the BP during 24 hours in an important number of patients. These characteristics can be detected by combining the office BP measurement, SMBP and ABPM. The alternative possibility would be lifestyle modification, appropriate drug combinations and to start treatment at lower levels than those currently used as thresholds (the guidelines for antihypertensive treatment have been drastically shifted in this direction over the past years). PMID- 12085420 TI - [Simultaneous presence of antibodies against the glomerular basement membrane and anti-myeloperoxidase antibodies in 2 patients with rapidly progressive glomerulonephritis]. AB - We report two patients with rapidly progressive glomerulonephritis without alveolar hemorrhage. Renal biopsy showed extracapillary glomerulonephritis with linear deposits of immunoglobulin G. Serologically anti-glomerular basement membrane antibodies (Ac AMBG) and ANCA anti-myeloperoxidase were present. All patients were treated with steroids, cyclophosphamide and plasma exchange. One patient needed dialysis, and other one died from a renal biopsy complication. We discuss the epidemiologic, pathogenic and prognostic aspects of this association. PMID- 12085421 TI - [Recurrent episodes of acidosis with encephalopathy in a hemodialysis program patient with short bowel syndrome]. AB - We present a case of a patient with short bowel syndrome in a hemodialysis program, with recurrent episodes of serious acidosis. The presence of a D-lactic acidosis peak secondary to bacterial overgrowth in the intestine was discovered during an acute episode of acidosis, with neurological affection. The detection of acidosis in predialysis measurements and the acute episodes of acidosis, made it necessary to administer bicarbonate to the patient and give him additional hemodialysis sessions. PMID- 12085422 TI - [Persistence of the left superior vena cava discovered during the implantation of a hemodialysis catheter]. AB - A persistent left superior vena cava (PLSVC) is the most common thoracic venous anomaly. It is a persistent remnant of the left anterior cardinal vein that usually disappears in early embryological development as a result of compression between the left atrium and the hilum of the left lung. If it is not associated with other congenital cardiac anomalies it is usually asymptomatic but has important clinical implications in some situations. In this article, we describe a patient with bilateral SVC identified on a chest radiograph by a haemodialysis central venous catheter passing through it. PMID- 12085423 TI - [Morbidity in hemodialysis]. PMID- 12085424 TI - [Uricemia-lowering activity of losartan in kidney transplantation]. PMID- 12085425 TI - [Acute renal failure and leukocytoclastic vasculitis as the presenting form of infectious endocarditis caused by Streptococcus bovis]. PMID- 12085426 TI - [Hormone therapy with estrogen patches for the treatment of recurrent digestive hemorrhages in uremic patients]. PMID- 12085427 TI - [Macroscopic hematuria after immunization with tetanus toxoid in a patient with IgA nephropathy]. PMID- 12085428 TI - [Latecoming to dialysis as a consequence of unidentified renal insufficiency]. PMID- 12085429 TI - [Importance of the residual renal function in hemodialysis patients]. PMID- 12085430 TI - Evidence-based practices: only the beginning. PMID- 12085431 TI - Managing to get by? PMID- 12085432 TI - Improving access to care at community mental health centers. PMID- 12085433 TI - Safeguards for research subjects: who's watching whom? PMID- 12085434 TI - Two worlds of treatment are converging. PMID- 12085435 TI - Moving services under one roof. Providers explore advantages of cross-training staff. PMID- 12085436 TI - What are the numbers? PMID- 12085437 TI - [Prevention of aging. Communication I. Individual enzymatic variations of the antioxidant system and a way to correct this system by means of electrochemically activated systems]. PMID- 12085438 TI - [Physiological mechanisms of respiratory phenomena in anxiety and depressive disorders]. PMID- 12085440 TI - [Influence of the direction of a moving target on reaction time in subjects of various ages]. PMID- 12085439 TI - [The problem of recovery of physiological functions in humans in deep accidental hypothermia (to the problem of limits of physiological adaptation)]. PMID- 12085441 TI - [Perfusion pressure dynamics in lower extremities at rest and after exercise]. PMID- 12085443 TI - [Contents of cadmium, lead, copper in blood and breast milk of mothers living in a region of ecological tension (as exemplified by cities of Balkhash and Karaganda)]. PMID- 12085442 TI - [Pituitary, adrenal and sex hormones in saliva]. PMID- 12085444 TI - [EEG correlations of emotional disorders and behavioral deviations in schoolchildren]. PMID- 12085445 TI - [Effect of psychophysiological potential on adaptation to educational activity]. PMID- 12085446 TI - [Changes in the EEG rhythms in endogenous depressive disorders and the effect of pharmacotherapy]. PMID- 12085448 TI - [Auditory perception of voice perception and abnormal sounding]. PMID- 12085447 TI - [Effect of individual personality on changes of sleep structure caused by emotional stress]. PMID- 12085449 TI - [Main factors of a decrease in stress resistance in six- to eight-year-old children with long-term consequences of perinatal CNS pathology during transition to school period of their life]. PMID- 12085450 TI - [Significance of individual features in interpreting the indices of spectral components of the cardiac rythm]. PMID- 12085451 TI - [New parameters for evaluating the functional state in patients with angina pectoris]. PMID- 12085452 TI - [Features of the application of heart rhythm variability analysis in cardiologic patients]. PMID- 12085453 TI - [The state of regional blood flow in the upper extremity after dosed physical exercise]. PMID- 12085454 TI - [Vibration-induced frontal postural reactions in humans standing on unstable supports of various types]. PMID- 12085455 TI - [Study of temperature homeostasis in real and simulated weightlessness]. PMID- 12085456 TI - [Physiological criteria of regulating the duration of physical work with submaximal power under conditions of limited heat dissipation]. PMID- 12085459 TI - Rage 2002. AB - Rage is the rage today. New phrases continue to make the headlines: sky rage, road rage, school rage, workplace rage, sports rage, parent sport observer rage, prayer rage, courtroom rage and more. The Internet advertises aggressive driver course: road rageous. PMID- 12085457 TI - [The methodology for assessment of the alpha-adrenergic sensitivity by the results of clopheline instillation into conjunctival sac in healthy subjects]. PMID- 12085458 TI - Treatment of osteoporosis: current status and recent advances. AB - With the advent of new treatment choices, osteoporosis should no longer be considered an inevitable part of aging but rather a disease that can be treated. This article provides an overview of recent advances in treatment of osteoporosis. PMID- 12085460 TI - Adolescent adherence in type 1 diabetes. AB - Adolescents with Type 1 diabetes have poorer glycemic control and more hypoglycemia than do adults. While physiologic reasons for these problems exist, the most common problems are failure to administer insulin, monitor glucose levels, and maintain an appropriate meal plan. PMID- 12085461 TI - Mild thyroid failure [subclinical hypothyroidism]: to treat or not to treat? AB - Most endocrinologists agree that if the serum TSH level is above 10 mIU/L, thyroxine therapy is indicated. For lower values of serum TSH, thyroxine therapy depends on the presence or absence of antithyroid antibodies and the associated clinical condition. PMID- 12085462 TI - The sick building syndrome: what is it when it is? AB - Sick building syndrome is a commonly applied diagnosis; often abused and misinterpreted to denote headaches, dizziness, fatigue and eye irritation associated with a building. PMID- 12085463 TI - Women's health: meeting the challenges of primary care in the age of managed care. AB - Female issues are neglected by male dominated healthcare, national medical meetings, and medical research. The 1st Annual Meeting of Pri-Med Midwest should dispel that notion. Over 5,000 healthcare professionals learned new information about women's healthcare. PMID- 12085464 TI - Over-the-counter sports supplements: what clinicians need to know. AB - Sports supplements have gained popularity, especially after endorsements by well known professional athletes. Since these supplements are thought of as being "natural", users often think of them as being safe. We discuss 4 popular supplements: creatine, androstenedione, dehydroepiandrosterone, and chromium. PMID- 12085465 TI - Ocular pathology in acquired immunodeficiency syndrome. AB - Gross and microscopic ocular findings were prospectively studied in 38 human immunodeficiency virus (HIV)-seropositive subjects undergoing postmortem examination. Pathologic lesions were detected in 27 patients (71%), with 67% of the abnormal findings detected only microscopically. PMID- 12085466 TI - A physician's guide to personal stress management. AB - Physicians are at increased risk for many of the adverse consequences of stress. This paper discusses common sources of stress and presents strategies for coping with them. Indications for more individualized treatment with a mental health specialist are also discussed. PMID- 12085467 TI - Pneumocystis carinii pneumonia in patients with and without HIV infection. AB - Advances in the prevention and treatment of Pneumocystis carinii pneumonia in HIV infected patients have led to a decrease in the incidence and improved outcomes. Pneumocystis carinii pneumonia continues to be problematic in non-HIV infected immunocompromised patients. PMID- 12085468 TI - Continuing medical education in JOEM. PMID- 12085469 TI - Occupational eye injury due to phototoxicity. PMID- 12085470 TI - The role of physical therapy in occupational low back injuries. PMID- 12085471 TI - Effectiveness of early physical therapy in low back musculoskeletal disorders. PMID- 12085472 TI - Localized neutrophilic dermatosis following welding burns. PMID- 12085473 TI - Diabetic firefighters. PMID- 12085474 TI - HIV and AIDS in the workplace. AB - Since the onset of the HIV epidemic, AIDS and HIV infection have presented tremendous challenges to infected individuals seeking to remain productive in the workplace, to employers coping with the special needs of such individuals, and to physicians who treat and counsel exposed or infected personnel. OEM physicians should strive to ensure that employers are familiar with legislation and guidelines protecting the rights of infected employees, and they should support rational workplace policies applying to employees with HIV infection or AIDS. When the potential for occupational HIV exposure exists, OEM physicians should ensure that adequate training around exposure prevention, triage, and treatment is provided. OEM physicians who treat individuals with occupational HIV exposures should involve themselves in institutional efforts to prevent exposures through the use of safer devices and procedures, and they should ensure that immediate and adequate clinical evaluation of exposures is available at all times. PMID- 12085475 TI - Exposure to secondhand smoke in the workplace: serum cotinine by occupation. AB - To examine workplace exposure to secondhand smoke by occupation, we analyzed data from The Third National Health and Nutrition Examination Survey (NHANES III) (1988 to 1994), a nationally representative sample of the noninstitutionalized population. The analysis was restricted to 4952 employed nonsmoking adults who reported no home exposure to cigarette smoke. Occupations were assigned to 40 groups and 7 categories. Among the categories, geometric mean serum cotinine (ng/mL) ranged from 0.09 for farming/forestry/fishing occupations to 0.22 for operators/fabricators/laborers (median, 0.16). The lowest values were observed among farmers and nursery workers (0.06) and the highest among waiters (0.47). Between 1988 to 1991 and 1991 to 1994, the overall geometric mean cotinine and the proportion reporting that they could smell smoke at work decreased significantly. In conclusion, workplace exposure to secondhand smoke varied by occupation, and decreases in exposure occurred between 1988 to 1991 and 1991 to 1994. PMID- 12085476 TI - Work disability in The Netherlands: data, conceptual aspects, and perspectives. AB - Data on work disability lack a standardized approach when defining the main variables. As a consequence, these data do not usually provide reliable information about health problems and related outcome measures. Examples of this unreliability are provided by the rather different approaches used in Dutch social security compared with the national statistics on disability. Several authors have tried to cope with this by introducing concepts that recognize disability as the multifaceted phenomenon it is and still provide reliable data. We recommend the development and global use of one concept so as to make data comparable. The recently presented International Classification of Functioning, Disability, and Health seems to be a sound base for further exploration of the possibilities for standardization in occupational health and social security. PMID- 12085477 TI - Occupational risk factors for small bowel carcinoid tumor: a European population based case-control study. AB - Small bowel carcinoid tumor (SBC) is a rare disease of unknown etiology but with an age-, sex-, and place-specific occurrence that may indicate an occupational origin. A European multicenter population-based case-control study was conducted from 1995 through 1997. Incident SBC cases between 35 and 69 years of age (n = 101) were identified, together with 3335 controls sampled from the catchment area of the cases. Histological review performed by a reference pathologist left 99 cases for study; 84 cases and 2070 population controls were interviewed. The industries most closely associated (a twofold or more odds ratio [OR]) with SBC, taking into account a 10-year time lag after exposure were, among women, employment in wholesale industry of food and beverages (OR, 8.2; 95% confidence interval [CI], 1.9 to 34.9]) and among men, manufacture of motor vehicle bodies (OR, 5.2; 95% CI, 1.2 to 22.4), footwear (OR, 3.9; 95% CI, 0.9 to 16.1), and metal structures (OR, 3.3; 95% CI, 1.0 to 10.4). The identified high-risk occupations with an OR above 2 were shoemakers, structural metal preparers, construction painters and other construction workers, bookkeepers, machine fitters, and welders (men). The OR for regular occupational use of organic solvents for at least half a year was 2.0 (95% CI, 1.0 to 4.2). Exposure to rust preventive paint containing lead was suggested as another potential occupational exposure (OR, 9.1; 95% CI, 0.8 to 107). This explorative study suggests an association between certain occupational exposures and SBC, but some of these associations could be attributable to chance. All findings should be regarded as tentative. PMID- 12085478 TI - The economic burden of lost productivity due to migraine headache: a specific worksite analysis. AB - Large, epidemiologic survey studies have established that migraine headaches affect approximately 6% of men and 18% of women in the United States and that the condition peaks during the prime working years (25 to 55 years of age). The consequent economic burden experienced by employers is substantial. The majority of this economic burden is realized by employers in terms of lost productivity, a combination of costs attributable to absenteeism and to lost productivity while on the job ("presenteeism"). Although large survey studies have produced estimates of national prevalence and have suggested substantial national costs, specific employers are rarely able to apply these projections to their specific workforce. Using demographic and payroll data available from a large financial services corporation with over 80,000 employees, this study used established prevalence data to estimate corporate costs stemming from migraine-related absenteeism and reduced on-the-job productivity to total at least $21.5 M and $24.4 M. In addition, a comparison of predicted prevalence and cost impact was conducted using a simpler and less costly health risk appraisal. This assessment proved to be a reliable tool in assessing prevalence of migraineurs in this corporation's workforce. Its use with a sample of 19,853 employees at this corporation produced prevalence rates of 7.7% of men and 23.4% of women, estimates closely comparable to those of national surveys. Suggestions are made regarding a corporate response to the substantial costs of lost productivity associated with migraine headache. PMID- 12085479 TI - Physical leisure-time activities and long-term sick leave: a 15-month prospective study of nurses' aides. AB - The objective of the study was to determine the relationship between physical leisure-time activities and the occurrence of long-term sick leaves in nurses' aides. Of 5563 Norwegian nurses' aides who were not on sick leave when they completed a questionnaire in 1999, 4744 (85.3%) completed a second questionnaire 15 months later. Brisk walks (odds ratio [OR], 0.78; 95% confidence interval [CI], 0.63 to 0.98), aerobics or gymnastics (OR, 0.71; 95% CI, 0.49 to 1.02), and other physical leisure activities (OR, 0.81; 95% CI, 0.66 to 1.00) for 20 minutes or more at least once a week predicted fewer long-term sick leaves (> 14 days), after adjustments for baseline health complaints, demographic and familial characteristics, former and current smoking, and work factors. In conclusion, regular physical leisure-time activity may be associated with a reduced risk of long-term sickness absence in nurses' aides. PMID- 12085480 TI - Promoting health in the construction industry? AB - The construction industry has long been regarded as a high-risk industry in relation to health and safety issues. The purpose of this research was to investigate the extent of occupational health and safety provision within the residential development sector of the construction industry in Britain. An additional aim was to gauge the sectors' attitudes toward health promotion. Data were collected using a self-completed questionnaire that was sent to 200 residential developers; a 46% response rate was achieved. The results revealed a high level of knowledge of health and safety legislation; occupational health provision in the sector is low, but a strong appreciation was demonstrated of the value of such services. There was a low level of health promotion policy development in relation to the areas of alcohol, smoking, and skin cancer. PMID- 12085481 TI - Fitness self-perception and Vo2max in firefighters. AB - Firefighters work at maximal levels of exertion. Fitness for such duty requires adequate aerobic capacity (maximum oxygen consumption [Vo2max]). Aerobic fitness can both improve a worker's ability to perform and offer resistance to cardiopulmonary conditions. Inactive firefighters have a 90% greater risk of myocardial infarction than those who are aerobically fit. Participants (101 firefighters) completed a questionnaire that asked them to rank their fitness level from 0 to 7; e.g., Level 0 was low fitness: "I avoid walking or exertion, e.g., always use elevator, drive whenever possible." The level of activity rating increased to Level 7: "I run over 10 miles per week or spend 3 hours per week in comparable physical activity." Each participant then completed two measures of Vo2max: a 5-minute step test and a submaximal treadmill test. There was no association between the firefighters' self-perception of their level of fitness and their aerobic capacity as measured by either step test or submaximal treadmill. Because of the critical job demands of firefighting and the negative consequences of inadequate fitness and aerobic capacity, periodic aerobic capacity testing with individualized exercise prescriptions and work--community support may be advisable for all active-duty firefighters. PMID- 12085483 TI - In-season physiological and biochemical status of reforestation workers. AB - Tree-planting demands sustained high work-output and is associated with high injury rates but has not been characterized previously. Data were collected from 10 male planters after 19 +/- 5 (T1) and 37 +/- 5 days (T2) of planting. One severe infection and one knee strain injury were noted. Loads carried were 32% +/ 5% body mass and heart rates were between 60% to 75% of maximum for 57% +/- 26% of the planting shift. A loss of 1.7% +/- 1.5% body mass occurred, although intake exceeded 5000 kcal/day. From 8% (T1) to 27% (T2) of postprandial blood glucose samples were found to be < 3.3 mmol/L. Further evidence of the cumulative stress was seen in increased levels of resting cortisol (428 +/- 163 to 741 +/- 103 mmol/L), norepinephrine (2.09 +/- 1.34 to 3.09 +/- 1.05 mmol/L), and creatine kinase (184 +/- 82 to 397 +/- 174 U/L) at T2. The postactivity neutrophilia and lymphophilia (absolute count, CD4+ and CD8+) observed at T1 were blunted at T2. The implications of mild hypoglycemia, including possible effects on cognitive and motor function, warrant further study. PMID- 12085482 TI - A case control study of lung cancer and non-malignant respiratory disease among employees in asphalt roofing manufacturing and asphalt production. AB - Two case control studies, one for lung cancer and one for non-malignant respiratory disease excluding influenza and pneumonia, of workers engaged in asphalt roofing manufacturing and asphalt production were performed to determine whether there was an increased risk associated with exposure to asphalt fumes or respirable crystalline silica in these industries. Industrial hygiene data for these roofing and asphalt plants do not exist before 1977. Pre-1977 exposure scenarios were constructed to estimate historic exposures for asphalt fumes and respirable crystalline silica. The only statistically significant elevated ORs were for cigarette smoking in both the lung cancer and the non-malignant disease analyses. The lack of an apparent dose-response relationship with exposure to asphalt fumes argues against an association between exposure to asphalt fumes at levels present in the industries reported on here and lung cancer and non malignant respiratory disease, excluding influenza and pneumonia risk. PMID- 12085484 TI - Hepatitis C in urban and rural public safety workers. AB - A sample of 719 Oregon public safety personnel (police officers, firefighters, and corrections officers) was tested for hepatitis C virus (HCV) antibody after completing a risk questionnaire. Seven of nine positive enzyme immunoassay tests (78%) were confirmed with recombinant immunoblot assay, yielding confirmed prevalence estimates of 1.2% (95% confidence interval, 0.4 to 2.8%) among the 406 firefighters and emergency medical technicians, and 0.7% (95% confidence interval, 0.1 to 2.6%) in 274 corrections personnel. No cases were observed in the 29 participating police officers. Self-reports of the number of workplace exposures to blood were not associated with HCV positivity, and the number of years of public safety employment seemed to be slightly less for HCV-positive subjects. Two of the seven (28.6%) HCV-positive individuals reported having at least one nonoccupational risk factor (odds ratio, 4.3; 95% confidence interval, 0.4 to 27.1), suggesting the greater relative importance of nonoccupational exposures. PMID- 12085485 TI - The interaction of education and cumulative lead exposure on the Mini-Mental State Examination. AB - The effect of lead exposure on neurobehavioral performance is modified by age. Whether educational achievement can serve as an effect modifier on the lead cognitive performance relationship is examined. The Mini-Mental State Examination (MMSE) and the reading section of the Wide Range Achievement Test-Revised (WRAT R), a measure of educational achievement, were administered to 256 lead smelter workers. The workers had a mean (standard deviation) age of 41 (7.9) years, education of 10 (2.8) years, employment duration of 17 (8.1) years, current blood lead of 28 (8.8) micrograms/dL, and working lifetime integrated blood lead index (IBL) of 725 (434) micrograms-yr/dL. The median (range) MMSE score was 29 (19 to 30). Multiple linear regression assessed the contribution of age, WRAT-R, education, alcohol intake, cigarette use, IBL, and IBL x WRAT-R on MMSE performance. A significant IBL x WRAT-R interaction examined by stratification found a significant dose-effect relationship between IBL and MMSE, but only in the 78 workers with a WRAT-R reading grade level below 6 years. Workers with higher educational achievement compensated for the effect of lead on cognitive performance. PMID- 12085487 TI - Prevalence and risk factors of occupational asthma among hairdressers in Turkey. AB - This study was designed to evaluate the questionnaire-based prevalence and possible risk factors of occupational asthma among hairdressers in Turkey. We investigated occupational history and respiratory, ocular, dermal, and nasal symptoms using a standardized questionnaire, evaluated worksite pulmonary function tests, and performed allergen skin testing. We then determined asthma risk factors using age- and gender-adjusted logistic regression models. The prevalence of occupational asthma in hairdressers was 14.6%. The odds ratio for hairdressers in a high work intensity group was 3.6 (95% confidence interval, 1.2 to 10.9) with a significant dose-response trend (chi 2 trend = 4.875; P = 0.027). The odds ratio for occupational asthma among workers with atopy was 4.5 (95% confidence interval, 1.2 to 17.2). We also observed an excess risk of occupational asthma with allergic rhinitis and conjunctivitis. Occupational asthma did not differ among subgroups of hairdressers. We observed an important risk of occupational asthma among hairdressers. The most prominent risk factors were work intensity and atopy. PMID- 12085486 TI - Exposure to asbestos and lung and pleural cancer mortality among pulp and paper industry workers. AB - We studied the mortality from lung and pleural cancers in a cohort of 62,937 male workers employed for at least 1 year in the pulp and paper industry in 13 countries during 1945 to 1996. Mill departments were classified according to probability and level of exposure to asbestos on the basis of available dust measurements and mill-specific information on exposure circumstances. Thirty-six percent of workers were classified as ever exposed to asbestos. Standardized mortality ratios of lung cancer were 0.99 (95% confidence interval [CI], 0.90 to 1.08) among unexposed and 1.00 (95% CI, 0.90 to 1.11) among ever exposed workers. The number of pleural cancer deaths among unexposed workers was 10; that among exposed workers was 14, most of which occurred among maintenance workers. In internal analyses, a trend in mortality from either neoplasm was suggested for estimated cumulative exposure to asbestos, weighted for the individual probability of exposure within the department and for duration of exposure (relative risk for lung cancer for 0.78+ f/cc-years, as compared with < or = 0.01 f/cc-years: 1.44; 95% CI, 0.85 to 2.45; corresponding relative risk for pleural cancer: 2.43; 95% CI, 0.43 to 13.63). Despite a possible nondifferential misclassification of exposure and outcome, this study suggests that the carcinogenic effect of asbestos can be detected among workers employed in industries such as the pulp and paper industry, in which it is not considered to be a major hazard. PMID- 12085489 TI - A physician's notes for community education in bioterrorism. PMID- 12085490 TI - Bioterrorism syndromes. PMID- 12085488 TI - Occupational exposures and risk of hepatitis B virus infection among public safety workers. AB - We conducted a questionnaire and seroprevalence survey to determine the frequency and type of occupational exposures (OEs) and the risk of hepatitis B virus (HBV) infection experienced by public safety workers (PSWs). Of the 2910 PSWs who completed the survey, 6.8% reported at least one OE in the previous 6 months, including needlestick (1.0%), being cut with a contaminated object (2.8%), mucous membrance exposure to blood (0.9%), and being bitten by a human (3.5%). The rate of OE varied by occupation with 2.7% of firefighters, 3.2% of sheriff officers, 6.6% of corrections officers, and 7.4% of police officers reporting > or = 1 OE (P < 0.001). The HBV infection prevalence was 8.6%, and after adjustment for age and race, it was comparable to the overall US prevalence and did not vary by occupation. By multivariate analysis, HBV infection was not associated with any OEs, but it was associated with older age, being nonwhite, and a previous history of a sexually transmitted disease. This study demonstrated that although OEs are not uncommon among PSWs, HBV infection was more likely to be associated with nonoccupational risk factors. Administration of hepatitis B vaccine to PSWs early in their careers will prevent HBV infection associated with occupational and non OEs. PMID- 12085491 TI - Bioterrorism threat makes preparation essential. PMID- 12085492 TI - The USA preparing for bioterrorism: the role of Fort Detrick since 1970. PMID- 12085493 TI - Creatine supplementation in Wisconsin high school athletes. AB - BACKGROUND: Creatine is a nutritional supplement used to enhance athletic performance in collegiate and professional athletes. There is increasing evidence that high school athletes are using creatine as well. The objective of this study was to describe patterns of creatine supplementation as well as the behaviors and beliefs associated with creatine use in high school athletes. METHODS: 4011 high school student-athletes from 37 public high schools in Wisconsin took part in a cross-sectional, multi-site, anonymous, descriptive survey. Measurements included self-reported patterns of creatine use. RESULTS: 16.7% of the athletes (25.3% males, 3.9% females) reported using creatine. Creatine use was lowest in the 9th grade (8.4%) and highest in the 12th grade (24.6%). The percentage of participants in each sport who used creatine varied considerably from 1.3% (female cross country) to 30.1% (football). Increased strength was the most likely perceived benefit of creatine supplementation, while dehydration was cited most often as a perceived risk of creatine use. Users were encouraged to take creatine most often by their friends while their parents discouraged its use. CONCLUSION: Despite the lack of research regarding the efficacy or safety of creatine supplementation in high school athletes, creatine was used by 25% of males and 4% of female high school athletes in Wisconsin. High school athletes who use creatine may not be aware of the risks and benefits associated with creatine supplementation. Primary care providers and sports medicine professionals need to educate athletes, coaches and parents about the creatine use as a performance enhancing supplement. PMID- 12085494 TI - CT screening for lung cancer: are we ready yet? AB - As a result of recent publications in the lay press, individuals have started to ask physicians and other medical personnel about the availability of screening CT for lung cancer. Many imaging centers, perhaps without full knowledge of the complex issues involved, have capitalized on this enthusiasm by instituting CT screening programs. Whether a health care provider offers screening CT or not, physicians and other medical personnel need to be informed about recent developments in CT screening for lung cancer in order to advise patients. This article reviews the significance of lung cancer as a cause of death, the causes of lung cancer and how it can be prevented, the types and frequencies of different lung cancers, the conditions of effective screening, the biases associated with screening, and past and present lung cancer screening trials. PMID- 12085495 TI - Preparing for bioterrorism. PMID- 12085498 TI - Breaking the mold. PMID- 12085497 TI - Inner turmoil. Prescription privileges make some psychologists anxious. PMID- 12085499 TI - Legal circumvention. PMID- 12085500 TI - Vox Populi. The voice of the people reveals why evolution remains controversial. PMID- 12085501 TI - Sweet medicines. PMID- 12085502 TI - Last mile by laser. PMID- 12085503 TI - The nose takes a starring role. PMID- 12085505 TI - Uncovering supersymmetry. PMID- 12085504 TI - The trials of an artificial heart. PMID- 12085506 TI - 15 answers to creationist nonsense. PMID- 12085507 TI - "The child that might be born...". PMID- 12085508 TI - Reprogenetics: a chance for meaningful regulation. PMID- 12085509 TI - Bioethics consultation in the private sector. PMID- 12085510 TI - Who will watch the watchers? PMID- 12085511 TI - Science, bioethics, and the public interest: on the need for transparency. PMID- 12085512 TI - Overseeing research on therapeutic cloning: a private ethics board responds to its critics. AB - Advanced Cell Technology's Ethics Advisory Board has been called window dressing for a corporate marketing plan. But the scientists and managers have paid attention, and the lawyers have gone along. PMID- 12085513 TI - Conception to obtain hematopoietic stem cells. AB - A couple may have a child to provide stem cells for another child. They may also use preimplantation testing--even, troubling though it is, prenatal testing and selective abortion--to ensure a close tissue match. PMID- 12085514 TI - Catholic views on life. PMID- 12085515 TI - What are families for? Getting to an ethics of reproductive technology. AB - The standard approach to the ethics of reproductive technologies starts and ends with the parents' procreative liberty. There's much more to think about. We should start with the relationship between parents and children. PMID- 12085516 TI - On a new charter to defend medical professionalism: whose profession is it anyway? PMID- 12085517 TI - Proper punishment. PMID- 12085518 TI - Wars of petition. PMID- 12085519 TI - Ethics review needs to grow up. PMID- 12085520 TI - The conscious incompetent patient. PMID- 12085521 TI - A new method of marking dentures using microchips. AB - Over the years various methods of denture marking have been reported in the literature. They include surface marking and inclusion techniques using metallic or non-metallic materials, microchips and microlabels. The microchips are preferred because of their small size and aesthetic acceptability. They are not however widely used due to the high cost of manufacture and data incorporation. This article details the procedures involved in inscribing a microchip using the photochemical etching process used in the electronics industry. The resulting microchip was cosmetically appealing, cost effective and was able to satisfy all the forensic requirements for a suitable denture marker. PMID- 12085522 TI - A case of bitten Bettongs. AB - Australia has a unique collection of native fauna, which is often threatened by physical harm or the destruction of its habitat and conservation of endangered species is a primary concern. Investigation of the recent deaths of Bettongs in Lincoln National Park, South Australia was undertaken by the Forensic Odontology Unit, Adelaide University to determine the likely perpetrator. Feral domestic cats were deemed to be responsible, as indicated by bitemarks on the radiotransmitter collars. PMID- 12085524 TI - Population studies of the Y-chromosome of loci DYS390, DYS391 and DYS393 in Brazilian subjects and its use in human identification. AB - Human identification can be accomplished by several technical procedures, especially by the comparative analysis of dental documents. Recent advances in molecular biology have now widened the scope for human identification and defined the protocols for the collection of biological material. Allele patterns of a population are particularly useful and they should be verified since they vary in different populations. We have studied the frequency of the alleles in three STR loci of the Y chromosome (DYS390, DYS391 and DYS393) in a group of Brazilian caucasian subjects. Results presented alleles 21, 22, 23, 24, 25 and 26 in locus DYS390, alleles 8, 9, 10, 11 12 and 13 in STR DYS391 and alleles in STR DYS393 were 11, 12, 13 14 and 15. The highest frequencies were 24 (0.46), 11 (0.37) and 13 (0.45). Each of the three STR systems had a PE (power of exclusion) of 0.6764 (DYS390), 0.5988 (DYS391) and 0.6136 (DYS393), The combination of the three STR systems revealed a PE of 0.9498, suggesting that this data can help in human identification. PMID- 12085523 TI - Comparison microscope identification of a cheese bitemark: a case report. AB - Police investigating the murder of a farmer recovered a piece of cheese containing bite- marks. The local dental practitioner used white plaster to make casts of the bitemarks in the cheese and also of the teeth of three suspects. The cheese specimen was retained by the police and seven months later the case was referred to the Forensic Odontology Unit at the University of Pretoria where a silicone rubber cast of the bitemarks in the cheese was made. A lack of concordant features present in a conventional pattern-associated comparison was overcome with the aid of a Leica DMC comparison microscope. Individual features observed under 6.3x magnification aided in the positive identification of the suspect, who when confronted with the evidence, admitted guilt at his first court appearance. PMID- 12085525 TI - Monogamy and its discontents: on winning the oedipal war. AB - This paper explores how idealized notions of "the beloved" constrain views of love and impede efforts to build satisfying relationships. The author begins by looking at psychoanalytic depictions of love. The effect of culture on beliefs about love is considered, exploring how cultural prescriptions, ideology, and group pressure (including psychoanalytic depictions of mature love) constrain and define views and values. Case vignettes and an example from literature illustrate ways in which cultural prescriptions obfuscate individual needs and impede self definition, thereby interfering with real intimacy and satisfaction in love relationships. PMID- 12085526 TI - Why insight fails. AB - Psychoanalysts rely on insight to promote therapeutic change. However, even when a great deal of insight has been attained, significant change may not be forthcoming in some patients, more often in borderline-level patients. The author examines such a case with particular reference to E. Kris' major points on insight, including the role of the ego's integrative functions in the attainment and utilization of insight. The author contends that failure to utilize insight is overdetermined and not necessarily owing to impairments in the integrative functions or to resistance in all its expressions. The author postulates that failure to make progress is due to a combination of the strength of the unmodulated drives, general ego weakness, poor early object relations, particularly preverbal experience, and constitution, in interaction. The whole of the personality is implicated in the inability to utilize insight. PMID- 12085527 TI - "Living turned inside out": the musical expression of psychotic and schizoid experience in talking heads' Remain in light. AB - The rock album form, advantageously suited for the artistic expression of intrapsychic experience, has been virtually ignored by analytic writers. Remain in Light by Talking Heads is presented as an example of an album that effectively utilizes the potentialities of this form to give powerful and disconcerning musical expression to a variety of psychotic and schizoid experiences, bringing to life the formulations for these phenomena of Bion, Winnicott, Guntrip, Ogden, Grotstein and others. The album suggests a variety of mechanisms by which sound and music may serve both defensive and compensatory functions in relation to these phenomena. PMID- 12085528 TI - Escaping the dark side of curiosity. AB - Curiosity as a clinical entity has been a neglected subject in the psychoanalytic literature. Freud never addressed the issue of curiosity systematically. His interest was in trying to account for children's sexual questioning. Nevertheless, hinderance to internal curiosity--this is to say, that which intimidates and abates the appetite for an exploration of one's motives--is part and parcel of psychoanalytic inquiry. And, arguably, there is no greater clinical challenge for the analyst than trying to treat an analysand who appears to lack an interest in the underlying causes of his unhappiness. The problem of impeded self-inquiry is usually exacerbated in people with more serious emotional disturbances. My position here is that in studying the conditions that mitigate against curiosity in a seriously disturbed patient, we gain access to an enlarged version of the curiosity problems of our less disturbed patients. Here I interpret my clinical impressions about problems with curiosity with ideas from the writings of Martin Buber and Albert Camus. PMID- 12085529 TI - Supportive psychotherapy for borderline patients: a psychoanalytic research perspective. PMID- 12085530 TI - Between wonder and doubt: psychoanalysis in the goal-free zone. AB - This paper examines the experiential dimensions of wonder and doubt as one way to articulate the creative and growth-promoting tensions between the medical/scientific model and the spiritual/mystical model. Both forms of experience, it is argued, function as necessary elements in a psychotherapy that integrates psychoanalysis and spiritual praxes. Fundamental differences between the medical/scientific model and the spiritual/mystical models are examined. The notion of the gap serves to illustrate these diametrically opposed, albeit compatible and necessary, points of view. The multifaceted layers of movement between Buddhism and psychoanalysis typify interacting dynamics between the two disciplines and provide a focal point for discussion. The paper then explores parallels between the Zen Buddhist notion of "Satori" as explicated in the writings of D. T. Suzuki and Wilfred Bion's notion of "O." Theoretical aspects of the discussion provide a backdrop for exploring clinical experience regarding the relationship between acceptance and change, the clinical relevance of the Buddhist notions of "gaining idea" and "basic goodness." The author also explores relationships between presence as actuality and ideal; knowing and not knowing; wonder and doubt. Clinical material supports the theoretical aspects of the discussion. PMID- 12085531 TI - [Diagnostic errors in nephrology]. PMID- 12085532 TI - [Diagnostic errors in gastroenterology]. PMID- 12085533 TI - [Acute posterior myocardial infarct. A typical diagnostic error]. PMID- 12085534 TI - [Misleading systemic manifestations of rheumatoid arthritis]. PMID- 12085535 TI - [Inflammatory thyroid diseases. Pathophysiology, diagnosis and therapy]. PMID- 12085536 TI - [30-year-old patient with progressive bone pain and muscle weakness. Adult form of phosphate diabetes with secondary hyperparathyroidism]. PMID- 12085537 TI - [A rare cause of recurrent hemoptysis. Unilateral pulmonary arterial hypoplasia as a cause of recurrent hemoptysis and bronchitis]. PMID- 12085539 TI - [The RENAAL Study. Effect of losartan on diabetic nephropathy]. PMID- 12085538 TI - [Progress in therapy of erectile dysfunction. New orally administered drugs]. PMID- 12085540 TI - [Progress and continuing education in medicine. Report on the 26th Interdisciplinary Forum of the Federal Physicians Council, 10-12 January 2002 in Cologne]. PMID- 12085541 TI - [Molecular Medicine 2002. Report of the satellite conference within the scope of the IX Gastroenterology Seminar Week 6 February 2002 in Titisee]. PMID- 12085542 TI - [PET scan versus MRI in patients with neuropsychiatric systemic lupus erythematosus. A negative PET scan is unusual]. PMID- 12085544 TI - Botany on a plate. Pleasure and the power of pictures in promoting early nineteenth-century scientific knowledge. AB - In early nineteenth-century Britain the use of pictures in introducing novices to the study of science was contentious, leading to debates over the ways in which words and images constituted knowledge and over the role of pleasure in intellectual pursuits. While recent studies have stressed visual representation as a critical element of science and considered its relation to the written word in conveying information, this essay explores the nineteenth-century preoccupation with the mind and mental faculties in relation to corporeal responses to explain concerns over the role of images and the process of recognition. By considering illustration in this way, it argues that popular botany was defined by many expert naturalists as the means by which private individuals could best be encouraged to extend their aesthetic appreciation and love of plants to an active and participatory pursuit of science. PMID- 12085547 TI - [Exercise intensity and fat burning--theoretical principles and practical considerations]. AB - There are several methods to eliminate unnecessary subcutaneous adipose tissue. Aerobic exercise seems to be the cheapest and most efficient method compared to diets, pills or even surgery. Subcutaneous adipose tissue can only be reduced by lipolysis and oxidation in the working muscles. The intensity of exercise is the most relevant factor in fat oxidation. Even though fat oxidation is highest at low to moderate intensities, well trained athletes have their highest fat oxidation at the ventilatory threshold. Therefore, different intensities for trained compared to untrained persons have to be recommended in order to burn fat. The determination of concentrations of lactate seems to be of important practical use. PMID- 12085548 TI - [Mechanisms of action of inhalation anesthesia: from Meyer-Overton into the new millenium]. PMID- 12085550 TI - [Abdominal pain of unknown origin with changed stool habits. Irritable bowel syndrome]. PMID- 12085549 TI - [22-year-old patient with left groin pain]. AB - A 22 year old man from Ethiopia suffered from progressive left inguinal pain and weight loss for ten months. The pain aggravated with leg movement, in particular with flexion of the left hip. ESR and CRP were slightly elevated, and a PPD was strongly positive. Abdominal ultrasound was normal but CT-scan revealed a left sided psoas abscess. Pott's disease was suspected and tuberculostatic therapy with INH, PZA, ETH, RIF was initiated immediately. A MRI of the spine excluded spondylodiscitis. Primary Tb psoas abscess was diagnosed. Treatment response after 5 weeks was clinically insufficient and CT-scan showed enlargement of the abscess. Treatment adherence was verified by drug prescriptions and INH urinary stix testing. M. tuberculosis, suspected microscopically in the puncture fluid, grew in culture and was fully drug sensitive. After 12 weeks, surgical abscess debridement had to be performed due to disease progression. The patient's health state improved considerably the first two postoperative months, inflammatory markers normalized, although a small residual abscess was still visible on CT. Subsequently, three months after surgery, pain reemerged, CT showed abscess progression. The patient had to be reoperated. Tb psoas abscess was a frequently described complication of Tb spondylodiscitis (Pott's disease) the first half of the last century and became rare thereafter in the Western hemisphere. However, the last two decades, due to migration policies and a worldwide increase of Tb epidemic because of socioeconomic destabilization and spread of the HIV-pandemic, Tb reemerged in Western countries. Therefore, physicians should be aware of atypical manifestations of tuberculosis. Primary Tb psoas abscess is extremely rare. Only four cases are described in the literature. In analogy to Pott's disease, therapy consists of tuberculostatic treatment, supported by surgical debridement. PMID- 12085551 TI - [50-year-old patient returning from Hawaii with changed stool habits. Blastocystis hominis]. PMID- 12085552 TI - [Patient questions on Barrett esophagus]. PMID- 12085553 TI - [Television, children and epilepsy]. AB - Two girls and one boy suffered seizures caused by television and other visual stimuli from 11, 12 and 12 years of age onwards, respectively. EEG recording revealed that intermittent photic stimulation (IPS) provoked epileptiform activity. Technological progress (video games, computer, disco, car, train) has considerably increased the risk for visually-induced seizures. A comprehensive clinical history with special attention to the environmental circumstances is important. For correct diagnosis an EEG with standardised IPS is necessary. Treatment consists of avoidance of strong visual stimuli. Patients may need prophylaxis with valproic acid, which should only be withdrawn after clear reduction of the EEG response to IPS. Repeating the EEG after the dosage has been lowered will help avoiding unnecessary recurrence of seizures. PMID- 12085554 TI - [Pharmaco-economic evaluations of new drugs: potential key to a more efficient allocation of the health care budget]. AB - In the light of rising expenditure on drugs and health care, a transparent, rational and careful decision-making process is required for the reimbursement of drugs. In the Netherlands, the Ministry of Health intends using pharmaco economics in this process, i.e., new drugs will not only be judged on their clinical efficacy but also on their cost-effectiveness. Guidelines for pharmaco economic research in the Netherlands have been published. According to these guidelines, a pharmaco-economic study must contain a cost-effectiveness analysis and/or a cost-utility analysis. In addition, a budgetary impact analysis is required. By 2005, all new drugs with therapeutic added value must supply a pharmaco-economic evaluation in order to apply for reimbursement. It will be the Minister of Health who decides whether a new drug will be reimbursed. PMID- 12085555 TI - [Haemangiomas and congenital vascular malformations: their classification and diagnosis]. AB - Mulliken and Glowacki's classification of peripheral blood- and lymph-vessel abnormalities is based on their clinical course and cellular characteristics, and is therefore clear to and readily usable by the practising physician. In order to make the diagnostic process more accessible, the Haemangiomas and Congenital Vascular Malformations Nijmegen working group has developed a system of diagnostic guidelines on the basis of this classification. The anamnesis should be directed at the following six distinguishing characteristics: presence of the anomaly at birth, growth, involution, change in volume, pain and outflow. The physical examination is directed at the following five characteristics: the possibility of emptying or pushing aside the anomaly, changes in volume during engorgement, murmur/'thrill'/pulsation, phleboliths, and hyper- or hypotrophy. If a diagnosis still cannot be made, then additional investigations may be carried out. Duplex scanning is usually sufficient for this purpose, after which the nature and extent of the malformation can be determined with MRI. On the basis of the results, the persons involved can be informed as to the prognosis of the malformation and a plan of treatment can be proposed. PMID- 12085556 TI - [Physical diagnosis--Ortolani's manoeuvre]. AB - Ortolani's manoeuvre is used to detect hip instability. It is intended to be used to establish the mobility of a dislocated femoral head with respect to the acetabulum in newly-born infants. Barlow's modification attempts to dislocate a reduced but unstable hip out of the acetabulum. Ortolani's manoeuvre is a primarily tactile, proprioceptive sensation and has nothing to do with the usually inconsequential audible and palpable clicks in infant hips. It has been found that widespread use of Ortolani's manoeuvre cannot prevent all late diagnoses of hip dysplasia. This is because neonatal hip instability is only part of the total spectrum of hip dysplasia, and because the examination requires a relaxed newly-born infant and an experienced examiner. In the Dutch situation, general screening with Ortolani's manoeuvre is not advisable as this will lead to many false-positive and false-negative outcomes due to this neonatal examination being carried out by various physicians and midwives with insufficient experience. Better results may be expected from ultrasound screening at child welfare clinics. This is presently being investigated in a large population study. PMID- 12085557 TI - [Diagnostic image (91). A collapsing man with an aortic aneurysm. Ruptured aneurysm of the abdominal aorta]. AB - A 73-year-old man had been diagnosed with an abdominal aortic aneurysm. Surgery was impossible due to poor cardiopulmonary condition. The patient collapsed due to rupture of the aneurysm. PMID- 12085558 TI - [Agreement between guidelines for pharmaco-economic research and never-before published health-economics evaluations]. AB - OBJECTIVE: To investigate whether the health-economics research published in Dutch journals is in agreement with the guidelines for pharmaco-economic research as published in 1999 by the Dutch 'College voor zorgverzekeringen' [Healthcare Insurance Board]. DESIGN: Descriptive. METHOD: A literature search was performed for health-economic evaluations published during 1990-1999 in Dutch journals such as Nederlands Tijdschrift voor Geneeskunde (NTvG), Pharmaceutisch Weekblad and Tijdschrift voor Gezondheidswetenschappen. The resulting 19 studies, primarily published in the NTvG, were compared with the pharmaco-economic guidelines, summarised in 9 criteria. RESULTS: The social perspective was taken in 5 studies (26%), non-related medical costs were excluded in 19 (100%), relevant subgroup analyses were performed in 16 (84%), effectiveness was explicitly differentiated from efficacy in 15 (79%), incremental analysis was performed in 18 (95%), an adequate period of analysis was chosen in 16 (84%), costs, benefits and health gains were discounted in 13 (68%), sensitivity analysis was included in the investigation in 10 (53%), and estimated cost prices were used in 9 (47%). CONCLUSION: The methodology in recently published Dutch health-economics research was partly in agreement with the later official guidelines for pharmaco-economic studies in the Netherlands. PMID- 12085559 TI - [Netherton's syndrome in two sisters]. AB - Two girls, sisters aged 4.5 years and 6 months, had experienced serious erythrodermia since birth, with scarcely any hair growth and they exhibited poor growth despite a hypoallergenic diet. On the basis of the dermatological condition ichthyosis linearis circumflexa and microscopic examination of a hair shaft in which trichorrhexis invaginata (bamboo hair) was observed, the diagnosis of Netherton's syndrome was established. In this autosomal recessive hereditary condition there is a defective production or maintenance of the stratum corneum. Apart from the skin and the hair abnormalities there is often an atopic constitution as well. The treatment consists of skin ointments and a high-energy diet due to the loss of protein via the skin. PMID- 12085560 TI - [One hundred years of the Association of Surgeons in the Netherlands. III. Gastrointestinal surgery]. AB - On 27 February 1989 the Nederlandse Vereniging voor Gastro-Intestinale Chirurgie [Netherlands Association for Gastrointestinal Surgery] was founded. The aim- improvement of quality by integrating scientific and clinical work--was already formulated 10 years previously by the Gastrointestinal Surgery Working Group. The integration proceeded carefully; the Netherlands Association for Gastrointestinal Surgery began as a chapter of the Association of Surgeons in the Netherlands and as a working group within the Dutch Gastroenterology Association. Meanwhile new techniques have been investigated and introduced and surgical oncology is a growing area of collaboration with the Dutch Association for Surgical Oncology. In the future minimally invasive interventions and robot surgery will come to the fore, reflux disease and inflammatory bowel diseases will decrease as an operative indication and early diagnosis will play a greater role. PMID- 12085561 TI - [One hundred years of the Association of Surgeons in the Netherlands. IV. Thoracic surgery]. AB - In the Netherlands, two disciplines are involved, in thoracic surgery. Firstly, general surgeons, since thoracic surgery initially consisted of surgical treatment for pulmonary tuberculosis, bronchiectasis and empyema. Once tuberculostatics became available, surgical treatment of pulmonary tuberculosis became less important and lung/thoracic operations were carried out primarily for lung cancer. Secondly, cardio-pulmonary surgeons (now known as 'cardiothoracic surgeons') also play a role in thoracic surgery. This discipline is also allowed to carry out heart surgery. Surgery for stage-I and -II lung cancer is the mainstay of the practice of general thoracic surgeons. Surgery after induction chemotherapy for stage-III tumours is under investigation to find out whether this will improve treatment results. Video-assisted thoracic surgery (VATS) is a most interesting development, and has led to the revival of volume-reduction surgery for lung emphysema. Education is of paramount importance in ensuring the quality of surgery. In the near future there will be a need for qualified thoracic surgeons in the Netherlands and organisational and financial measures should be taken to prevent a shortage. PMID- 12085562 TI - [A pilot study of hypovitaminosis D in apparently healthy, veiled, Turkish women: severe vitamin D deficiency in 82%]. AB - A pilot study was performed in March 2001 in order to estimate the prevalence of hypovitaminosis D in veiled women in the Netherlands. In a group of 51 Turkish women aged 14-63 years, 42 (82%) were severely deficient (25-hydroxyvitamin D (25 (OH)D) < 20 nmol/l), and 4 (8%) were moderately deficient (25(OH)D: 20-30 nmol/l). About half of the deficient women complained of muscle pain, muscle weakness or fatigue. These results confirm the presence of a serious public health problem with regard to vitamin D amongst veiled women. PMID- 12085563 TI - [Class substitution of drugs--between cost reduction and the position of the professionals in health care]. PMID- 12085564 TI - [Lost in the methodology (final). XXXIX. The ultimate truth]. PMID- 12085565 TI - [The standard, "Anemia in the primary midwife practice," issued by the Royal Dutch Organisation of Midwives: a risk of not recognizing iron deficiency and hemoglobinopathy]. PMID- 12085567 TI - The effects of chemoprophylaxis on acquired immunity against gastrointestinal nematodes in cattle. AB - The level and duration of exposure to gastrointestinal nematode infections are of crucial importance for the development of acquired immunity in first season grazing calves. An excessive reduction of host-parasite contact by chemoprophylaxis, pasture management, or both, causes a diminished level of acquired immunity. Moreover, the level of acquired resistance is negatively related to the degree of suppression of host-parasite contact. Whether or not a reduced resistance against establishment and development of gastrointestinal nematode infections has a negative effect on weight gain in the second grazing season, depends both on the intensity of the prophylaxis used and on the level of the challenge infection. Cross-sectional serological surveys and questionnaire surveys in The Netherlands, Germany and Belgium indicated that parasitic nematode control in first season grazing calves tends to be overprotective. Possible consequences of over-treatment, besides higher treatment costs and a reduced level of acquired immunity, are more drug residues in animal products and in the environment and increased selection for anthelmintic resistance. PMID- 12085566 TI - Chemokines and proteinases in autoimmune diseases and cancer. AB - Chemotactic cytokines or chemokines form a family of proinflammatory proteins that are functionally linked to various classes of proteases, including matrix metalloproteinases (MMPs). Both families of molecules are key players in the migration of inflammatory cells in autoimmune diseases and in invasive cancers. For example, the chemokine interleukin-8 acts as a fast secretagogue of gelatinase B in granulocytes and is increased in the synovial fluid of arthritis patients and may locally recruit and activate neutrophils. The latter are the most abundant inflammatory cell type in the joints of patients with rheumatoid arthritis. In the case of the inflamed joint, the contribution of matrix remodeling enzymes in the breakdown of cartilage and bone is trivial. Gelatinase B (MMP-9) was documented in autoimmune diseases and cancer by immunohistochemistry with the use of monoclonal antibodies. Studies in rheumatoid arthritis and multiple sclerosis led us to postulate the "Remnant Epitopes Generate Autoimmunity" or REGA model for autoimmunity. This model is based on the pathophysiological role of three major classes of molecules involved in aspecific primary immune defense mechanisms: the cytokines, the chemokines and the proteases. The REGA model has proven to be useful for the development of disease treatment strategies. Particular cytokines are disease-limiting and may thus be used for the treatment of autoimmune disorders. Cytokines and chemokines that induce enzymes promote disease and may be antagonized. Along this line of research, we have recently identified natural and biosynthetic chemokine antagonists. Some of these have shown potent antiviral activity against human immunodeficiency virus. It is expected that these might also become useful in the treatment of autoimmune diseases and invasive cancers. A similar effect may be expected by the antagonization of damaging proteases or with the use of recombinant or synthetic enzyme inhibitors. PMID- 12085568 TI - [Intestines and joints: an entity?]. AB - The intestinal inflammation and the inflammatory arthropathy in Crohn's disease and in spondyloarthropathy are tightly linked. Both forms of inflammation occur in clinical and subclinical forms in both diseases: 60% of patients with Crohn's disease have clinical and/or radiological signs of articular involvement--65% of patients belonging to the concept of spondyloarthropathy have endoscopic and/or histological signs of gut inflammation. The macrophages and the lymphocytes play a role either by the transport of bacterial antigens by macrophages from the gut to the synovium, either by the recirculation of activated lymphocytes attracted to the tissue by adhesion molecules. In both diseases, TNF alpha play a central role resulting in a rapid and spectacular improvement of the intestinal and articular inflammation by the administration of a monoclonal chimeric antibody directed against this cytokine (Infliximab). This clinical effect is associated or secondary to a restoration of the production of proinflammatory cytokines. According the importance of genetic factors in the etiopathogenesis of both diseases (mutation in the NOD2 gene for Crohn--HLA-B27 phenotype for spondyloarthropathy) future studies are necessary to find a common alteration explaining the link between gut and joint. This knowledge can help in the elucidation of the origin of both diseases. PMID- 12085583 TI - Noninvasive monitoring of human consciousness by near-infrared spectroscopy (NIRS) during high +Gz stress. AB - Noninvasive monitoring of the relative change in oxygen saturation (rSO2) in cerebral tissue by near-infrared multi-wavelength spectroscopy (NIRS) was investigated in humans under high acceleration (+Gz) stress. These profiles included sustained 15-second +Gz plateaus and repeated short duration +Gz pulses of varying duration. The end points in this study were loss of consciousness due to high +Gz exposure (GLOC). In many cases subjects demonstrated cognitive and physical symptoms related to reduced cerebral blood flow without frank unconscious, which has been called Almost Loss of Consciousness (ALOC). Both the rSO2 levels during and after the +Gz exposures and the total time subjects were incapacitated after GLOC were recorded. It was found that while the drop in rSO2 at the onset of GLOC was lower during pulse exposures as compared to sustained exposures, the total time to recovery from GLOC was longer during the sustained runs. By applying a better understanding of the nature and timing of +Gz-induced changes in cerebral tissue oxygenation, more efficient control systems for personal protective gear for pilots of high performance aircraft can be implemented. PMID- 12085584 TI - Investigating initial cell-seeding density and culture period of fibroblast growing on biodegradable tricalcium phosphate lysine disks. AB - "Tissue engineering" uses implanted cells, scaffolds, DNA, protein, and/or protein fragments to replace or repair injured or diseased tissues and organs. Recently, biocompatible materials, such as hydroxyapatite and tricalcium phosphate, have been associated with cellular migration, exposure, and an increase in inflammatory mediators. Complications are less likely after the implants have become integrated. A model was sought to study the influence of multiple cell densities on the rate of fibroblast in-growth into porous implants. Fibroblasts were seeded at three different densities (50, 100, 200 x 10(6) cells/well) onto the surface of a 24 well tissue culture plate in the presence of porous tricalcium phosphate lysine ceramic disks. Cell number, viability, and morphology were evaluated after 1, 2, and 3 days. The results show a linear 2 fold increase in cell number during the first two days in culture. By day 3, the cells initially seeded at 200 x 10(6) cells no longer exhibited linear growth. This suggests that the tissue culture wells were confluent. Interestingly, the cells growing in wells containing TCPL disks showed slightly more cell numbers than cells growing on plastic alone for each density. The results also revealed no significant differences between the groups for parameters of cell function such as protein concentration and markers of cellular damage. Evaluation of cellular morphology indicated changes in cellular appearance with time in culture. Overall, initial cell seeding is important for faster in-growth, but with time the cells seeded at lower cell numbers will also become confluent. In conclusion, this study provides a novel approach for the study of fibroblast in growth into implants with time. This work is important for understanding functional tissue engineering and ultimately result in safer and more efficacious repairs and replacements for the surgeon and patient. PMID- 12085585 TI - Accelerometer-based system for the detection of lameness in horses. AB - Video-based kinematic analysis of gait in horses is accurate for quantification of lameness and reliable for identification of the affected limb. Algorithms for the measurement of the vertical head and pelvic displacement and phase correlation with vertical displacement of one forelimb and hindlimb foot have been developed for this purpose. However, because of camera field-of-view limitations, video-based analysis of gait can only be reliably accomplished with the horse constrained to move on a treadmill. This paper describes the use of 2 single-axis accelerometers and 2 gyroscopic transducers as a measurement system for the identification and quantification of forelimb and hindlimb lameness in horses. Vertical head and pelvic acceleration are converted to displacement, lameness is quantified from previously developed algorithms, and affected limb is determined by correlation of head and pelvic signals with gyroscopic signals from the right forelimb and hindlimb feet. Signals from the 4 transducers are telemeterized at 200 Hz and collected to a receiver connected to a lap top computer, freeing the horse from the constraints of a treadmill laboratory setting. In this paper we describe the reliability of this new accelerometer based system in horses with induced lameness while trotting on a treadmill and freely outside overground. PMID- 12085586 TI - The operating version of the Eye Tracker, a system to measure saccadic eye movements. AB - The operating version of the Eye Tracker, a transducer and system using a technique to bounce infrared light off the eye to measure saccadic eye movements in any X-Y position is presented in this paper. Discussed is the method of reading and analyzing eye movement data using a 24-channel infrared optoelectronic array and computer algorithms that utilize a linear regression model to interpret and determine eye location, the 24-channels used to ensure accurate reading of eye position. Accuracy is also maintained by a signal processing system that attenuates incident light as well as ambient light. Also discussed is a novel method of mounting the infrared array on hemispherical shaped eyepieces that in turn are mounted on goggles styled after an ophthalmologist's test frames that is comfortably worn and adjustable in size to fit any subject. A computer controlled, wall mounted light bank facilitates targeting for eye movements. The Eye Tracker is built to meet standards of a professional medical device manufacturer following typical mechanical, electrical, and safety techniques unique to device packaging. PMID- 12085587 TI - A microcomputer-based respiratory and activity recording system. AB - A respiratory and activity recording system has been developed for monitoring health conditions and living patterns, such as activity/rest time periods and general activity level. The system employs a piezoelectric sensor, a low-power operational amplifier, a low-power 8-bit microcomputer and a 512 KB EEPROM. The piezoelectric sensor, whose electrical polarization voltage is produced by mechanical strain, records body movements produced by respiration, heart pulse, walking and running. The high and low frequency components from the recorded body movements are discriminated by high and low pass filters. The high frequency components reflect the body movements produced by cardiac vibrations, walking and running activities. The low frequency components are mainly generated by respiration. The degree of activity is obtained by summing the high frequency components for 1 second. The microcomputer stores the obtained low frequency components and activity to the EEPROM at 0.2 second and 1 second intervals, respectively. After recording, these stored data are downloaded to a desktop computer. The computer then detects behavior patterns from the activity data and respiration from the low frequency components. PMID- 12085589 TI - The digital hearing aid, wearable computing, and electrophysiological response. AB - Presbycusis is the most common form of hearing loss caused by aging and long-term exposure to sound energy. This type of ailment decreases the ear's ability to perceive high frequencies and localize sound, thus making comprehension more difficult. To compensate for this loss, the choice of a digital hearing aid has become more common. However, most digital hearing aids do little more that their analogue predecessor's by providing a single, fixed hearing response. Such a fixed response is not suitable for all of a user's auditory environments and typically causes a more rapid loss of hearing. Significant advancement in processing power and reduction in size of computing hardware has produced increasingly more powerful, more portable, and more personal computing devices. These advances have spurred research and development of wearable computing devices towards integrating both man and machine. By definition, the digital hearing aid is a wearable computing device. The development of a digital hearing aid with increased onboard processing that is aware of its owner's electrophysiological and auditory environments is an obvious progression. This awareness will give the hearing aid the ability to autonomously modify its own parameters to improve audibility and comprehension. Electrophysiological signals can be classified as naturally occurring or voluntarily controlled. Employing these signals will allow the hearing aid to adapt to its owner's external and internal stimuli. Research and initial experiments into the monitoring and use of electrophysiological response for the control of the digital hearing aid shall be presented. PMID- 12085588 TI - Biologically based machine vision: signal analysis of monopolar cells in the visual system of Musca domestica. AB - Machine vision for navigational purposes is a rapidly growing field. Many abilities such as object recognition and target tracking rely on vision. Autonomous vehicles must be able to navigate in dynamic enviroments and simultaneously locate a target position. Traditional machine vision often fails to react in real time because of large computational requirements whereas the fly achieves complex orientation and navigation with a relatively small and simple brain. Understanding how the fly extracts visual information and how neurons encode and process information could lead us to a new approach for machine vision applications. Photoreceptors in the Musca domestica eye that share the same spatial information converge into a structure called the cartridge. The cartridge consists of the photoreceptor axon terminals and monopolar cells L1, L2, and L4. It is thought that L1 and L2 cells encode edge related information relative to a single cartridge. These cells are thought to be equivalent to vertebrate bipolar cells, producing contrast enhancement and reduction of information sent to L4. Monopolar cell L4 is thought to perform image segmentation on the information input from L1 and L2 and also enhance edge detection. A mesh of interconnected L4's would correlate the output from L1 and L2 cells of adjacent cartridges and provide a parallel network for segmenting an object's edges. The focus of this research is to excite photoreceptors of the common housefly, Musca domestica, with different visual patterns. The electrical response of monopolar cells L1, L2, and L4 will be recorded using intracellular recording techniques. Signal analysis will determine the neurocircuitry to detect and segment images. PMID- 12085590 TI - A system for monitoring heart pulse, respiration and posture in bed. AB - A non-invasive system has been developed to monitor cardiac vibrations, respiration and posture of inbed hospitalized patients and elderly people who need constant care. These physiological variables are recorded by four 2 x 28 cm piezoelectric film acceleration sensors, eight 2 x 2 cm small pressure sensors and a 5 x 100 cm long pressure sensor. The piezoelectric sensors, attached to the chest over the heart during bed sleep or rest, detect the movements produced by the heartbeat and respiration. The eight small pressure sensors are attached at various positions on the upper and lower body. A longer pressure sensor, to detect the patient leaving the bed, is attached to the side of the bed. These sensor outputs are digitized at a sampling rate of 200 Hz using a 12-bit A/D converter and stored on a personal computer. The computer detects cardiac vibrations and respiration from upper chest movements and posture from the pressures recorded by small and long pressure sensors. PMID- 12085591 TI - Development of a PC-based system for recording flash electroretinograms in experimental animals. AB - This report presents the results of a student design project, the goal of which was to develop a user-friendly PC-based system for recording flash electroretinograms (ERGs) in experimental animals. This system consists of an LED stimulator with associated driver circuitry, a differential amplifier, and a LabVIEW-based software package that controls data acquisition and handling. The system was designed to be applicable to multiple animal models, and is currently being tested on both rats and horseshoe crabs. The electroretinogram is the summation of electrical responses from both neuronal and non-neuronal cells in the retina in response to stimulation by a flash of light. In this system, ERGs are recorded using DTL-fiber electrodes stretched across a moistened cornea, and referenced to cornea of the opposite, unstimulated eye. ERG signals are differentially amplified and digitized using a National Instruments data acquisition (DAQ) board. ERGs are elicited by a LED stimulator that is controlled by the software package through a driver circuit that limits current flow to the LED. A diffuser is located in front of the LED to ensure uniform illumination of the retina. Stimulus intensities are controlled by alteration of the duration of excitation of the LED by the TTL output of the software. The software package allows the user to customize stimulation and sampling settings. It also functions as a digital oscilloscope to display responses as they are obtained from the subject. This package also incorporates the ability to average signals, which is necessary for increasing signal-to-noise ratios when recording small ERG signals. PMID- 12085592 TI - Stress state during fixation determines susceptibility to fatigue-linked biodegradation in bioprosthetic heart valve materials. AB - Mechanical loading contributes to the structural deterioration of bioprosthetic heart valves. The influence of stress state during fixation may play a substantial role in their failure, linking fatigue damage caused by buckling and tension and the enzymatic degradation of glutaraldehyde-crosslinked collagen. Bovine pericardia were obtained immediately postmortem and 100 mm x 15 mm samples were cut in the base-to-apex direction. Half the samples were subjected to a uniaxial tensile stress of 250 kPa and half remained unloaded during a crosslinking treatment in 0.5% glutaraldehyde. Tissue samples were rinsed and cut into 16 mm x 4 mm test strips. Half of these strips were exposed to cyclic compressive buckling and alternating tension at 30 Hz for 20 million cycles (approx. 7.5 days) using a custom-built multi-sample fatigue system. Fatigue damaged and non-damaged samples were subsequently incubated at 37 C for 48 hrs in: (i) Type I bacterial collagenase (20 U/ml) buffered in 0.05 M Tris, 10 mM CaCl2 2H2O (pH 7.4) or (ii) 0.05 M Tris buffer (pH 7.4) only. In both cases, the samples were loaded sinusoidally between 40 and 80 g using a previously described microtensile culture system. Tissue removed from the bath was rinsed in 0.1 M EDTA solution and mounted in a servo-hydraulic mechanical testing system (MTS). Ultimate tensile strength (UTS), maximum tissue modulus, and fracture strain were determined. The percent collagen solubilized was assessed by a colourmetric hydroxyproline assay of the enzyme bath and tissue sample. All data were analyzed by analysis of variance (ANOVA). The results confirmed the synergy between fatigue damage and collagenase proteolysis in these materials; however, there were no significant differences in this effect between simple fixation and stress fixation up to 20 million cycles. There were significant decreases in the mechanical properties and an increase in the amount of collagen solubilized with increased exposure to fatigue cycling. PMID- 12085593 TI - Cytopathological changes in early stages of benign prostatic hyperplasia upon exposure to sustained delivery of androgens. AB - The objective of this study was to assess the possible morphological changes that occur in the columnar epithelial cells of the prostate peripheral zone during early development of benign prostatic hyperplasia. In this study benign prostatic hyperplasia (BPH) was encouraged by the continuous delivery of androgens using TCPL drug delivery devices. A total of 16 adult male rats were randomly divided into four groups (n = 4). Group 1 served as an intact control and Groups 2-4 were implanted with tricalcium phosphate lysine (TCPL) capsules designed to deliver continuous physiologic (40 mg) doses of specific androgens as follows: Group 2, testosterone (Test); Group 3, dihydrotestosterone (DHT); Group 4, androstenedione (AED), respectively. Upon completion of the study, the columnar epithelial cells were targeted for morphometric analysis. The means number of cells per high power field, cell length, cell area, nuclear area and nuclear to cytoplasmic ratio were measured using image analysis techniques. The results of this study showed that (1) the number of cells per high power field were not changed with treatment, (2) the cell length was decreased with treatment of all androgens, (3) the cell area was decreased with treatment, (4) the nuclear area was increased with treatment of Test and DHT, and (5) the N/C ratio was increased with all three androgens. These results suggest that very early in the development of BPH, remarkable changes occur in the nucleus of the columnar epithelial cells. These changes may indicate a specific physiological response to irritation arising from the continuous delivery of androgens. PMID- 12085594 TI - Dynamic thermomechanical studies on collagen denaturation: a comparative study of bovine pericardium and chordae tendinae. AB - Denaturation of the collagen molecule is a kinetic process, and hence a function of both temperature and duration of heating. Simple isothermal tests have traditionally provided the easiest way to isolate the role of heating in denaturation. In the present experiments, a Dynamic Hydrothermal Isometric Tension (DHIT) system has been custom-built and used to impose near-step changes temperature as a means to studying the dynamics of denaturation in fresh and crosslinked bovine Pericardium (PC) and Chordac Tendinae (CT). Irrespective of their treatment, both PC and CT followed the same pattern of force rise following a step in temperature. The force-time data were well-fitted using the Levenberg Marquardt nonlinear least-squares method and 3 exponential functions with unique time constants (tau 1, tau 2, tau 3). This suggests the presence of three discrete mechanisms in the denaturation process. The tau 1 values for PC and CT were significantly shorter than either tau 2, or tau 3. Also, the tau 1 values of the untreated groups (either PC or CT) were similar and significantly different than those for the crosslinked tissues. This indicates that crosslinking dramatically slows the dynamic denaturation in both tissues (identified with tau 1) to the same extent. This is of considerable interest since each tissue has unique and distinct collagen architecture; therefore the dynamics of denaturation depend more on the collagen molecule than on fibrous architecture. This is the first study to identify discrete mechanisms leading to isometric denaturation changes in two collagenous tissues. While it is believed that the mechanisms identified represent (i) progressive hydration and (ii) denaturation of collagen structure, more precise identification of the processes remain to be done. PMID- 12085595 TI - Quantitative characterization of rat tendinitis to evaluate the efficacy of therapeutic interventions. AB - Tendinitis is a painful soft tissue pathology that accounts for almost half of all occupational injuries in the United States. It is often caused by repeated movements and may result in loss of work and income. Current treatments for tendinitis are aimed at reducing inflammation, the major cause of the pain. Although anti-inflammatory drugs and various alternative therapies are capable of improving tendinitis, there are no quantitative scientific data available regarding their impact on inflammation. The objective of this study is to determine the time course for healing of rat tendinitis without intervention to be able to assess the efficacy of tendinitis treatments. We are interested in evaluating the therapeutic use of pulsed electromagnetic fields (PEMFs), a therapeutic modality that has been found to be beneficial for healing soft tissue injuries. Tendinitis was induced in Harlan Sprague Dawley rats by collagenase injections into the Achilles tendon, and tendons were collected for four weeks post-injury. To determine the amount of edema, we used caliper measurements of the rat ankles and quantified the tendon water content. To determine the extent of inflammation, we estimated the number of inflammatory cells on histological sections applying stereological methods. The data reveal that edema is maximal 24 hours after injury accompanied by a massive infiltration of inflammatory cells. Inflammatory cells are then gradually replaced by fibroblasts, which are responsible for correcting damage to the extracellular matrix. This natural time course of tendon healing will be used to evaluate the use of PEMFs as a possible therapeutic modality. PMID- 12085596 TI - Radial compliance of natural and mock arteries: how this property defines the cyclic loading of deployed vascular stents. AB - There have been a disturbing number of unexpected mechanical failures in deployed vascular stents and stent grafts. An analysis of the mechanical properties of mock arteries used to carry out standard durability testing on the aforementioned medical products suggests that some of the problems might be related to a frequency dependent change in the properties of these recipient vessels. This could lead to a lower than expected level of loading and an attendant reduction in severity of testing. A clear understanding of how a pulsating artery cyclically loads a deployed stent is important before a mock artery can be designed to carry out biologically relevant in vitro durability testing. In addition, knowledge of the time dependent response of both the vessel and the stent/stent graft is critical before any accelerated testing protocol can be properly designed. This paper will present an analysis of the loading on a stent versus pressure and diameter of the recipient vessel and how this varies with frequency. Actual compliance versus frequency data will be used from silicone mock arteries. PMID- 12085597 TI - Effect of TCP sintering temperatures on MRC-5 fibroblast proliferation and viability. AB - Calcium/Phosphate ceramics have recently been used as orthopaedic implants, drug delivery systems, and models for studies on bone remodeling. Although their use has been very successful, problems such as aseptic loosening, coagulation, fibrous tissue formation, and inflammation have been noted. The objective of the study was to investigate the effects of tri-calcium Phosphate (TCP) ceramics sintered at different temperatures on the proliferation of MRC-5 fibroblasts. This was accomplished by light microscopy morphologic analysis, measurements of MDA (a measure of cell damage) and total protein, and cell counts. The experimental design consisted of four experimental groups and one control group. 96 TCP ceramics were divided into 4 groups of 24, with each group being assigned a different sintering temperature of 800, 1000, 1200 or 1400 degrees celcius. Each temperature group was then further divided into 8 ceramics per phase of 24, 48 and 72 hours. All data were analyzed using Sigma Stat software. The data collected from this study provided useful information pertaining to the optimum manufacturing conditions that produce high proliferation rates with minimal cellular damage. At 72 hours (the phase simulating chronic conditions), cell counts and MDA analysis showed that the cells incubated with the TCP sintered at 1200 degrees C had the greatest proliferation and least damage when compared to the other groups. Morphological, MDA and protein analysis showed the cells incubated with the 1400 degrees C TCP capsules to have undergone lysis or other extreme cellular alterations. Overall, the data collected revealed a marked alteration in cellular function and morphology over the range of sintering temperatures that were tested. PMID- 12085598 TI - Bond strength of fibrin glue between layers of porcine small intestine submucosa (SIS). AB - This study investigated the strength of the bond between layers of small intestine submucosa (SIS, Cook Biotech, Inc., West Lafayette, IN) glued with commercially available fibrin glue (Haemacure Corporation). To determine the conditions leading to the highest bond strength, three parameters were varied: the concentration of the fibrin component, the concentration of the thrombin component, and the type of applicator used to apply the two components. Five glue concentrations and two applicator types, a Paasch Airbrush and one provided with the Haemacure glue kit, were studied. To make the test specimens, two pieces of SIS were each sprayed separately with 1 mL of one of the glue components. The two pieces were then adhered and allowed to cure for two minutes. After the panels were glued, frozen, and lyophilized, they were cut to size according to ASTM Standard D 1876: Peel Resistance of Adhesives (T-Peel Test). The panels were then rehydrated, and tests were performed in an MTS tensile testing machine set to pull at a constant rate of 1 mm/sec over a 100 mm span. The mean force over the duration of the test was computed as specified in the ASTM standard. The airbrush was found to produce a stronger bond than the applicator supplied by Haemacure. Judged qualitatively, the airbrush also produced a much more uniform spray and consistent flow rate than the glue manufacturer's applicator. The data suggest that a decrease in concentration of both glue components yields increased bond strength, although variability in the results also increased with decreased glue component concentration. PMID- 12085599 TI - Proliferation and morphological transformation of RMK cells exposed to hydroquinine containing ionomers. AB - Recent research in our laboratories has been directed towards the development of ionomeric polymers and monomers for use in biomedical applications such as adhesives, drug delivery matrices and tissue scaffolds. The chemical Hydroquinone (HQ) aids as a stabilizer and represents a major component in the development of the ionomers. However, hydroquinone in high concentration has the potential to initiate carcinogenic effects on cells. The curing reactions are based on free radical chemistry that require a radical scavenger, ascorbic acid (Asc) to adjust working and setting times and shelf-life stability. The few studies published on HQ have suggested that high dosages of HQ may stimulate apoptosis as well as an increased cellular leakage, however the effect of HQ on the biocompatability is unknown. Therefore the objectives of this study were to measure the functional capacity, cell proliferation and structural integrity of Rhesus monkey kidney epithelial (RMK) cells exposed to ionomer formulations containing 4 different levels of HQ. A total of 90 tubes of RMK (40,000 cells per tube) cells were divided equally into five equal groups. Group I served as a control and group II V were subjected to ionomers containing 0, 500, 1000, and 2000 ppm HQ. Cell numbers, morphology, cellular and supermatant MDA levels, and total protein analysis were performed. The results suggest: (I) All ionomer groups increased cellular proliferation except for the 2000 ppm HQ group, (II) MDA levels were increased in cells containing 2000 ppm HQ at 24 hours; and 0 ppm at 48 hours. It may be concluded that HQ concentrations over 1000 ppm may adversely affect biocompatability. PMID- 12085600 TI - Interrelationship between sex steroid hormones and cardiovascular disease using MRC-5 cell line as a model. AB - Increasing evidence reveals that low-density lipoproteins (LDL) and gender are significant risk factors in the development of cardiovascular disease (CVD). The long-term objective of this investigation is to determine the possible relationship between sex hormones, LDL, and the development of CVD. The specific aims of this investigation include: (1) to evaluate the effect of estrogen (E) and testosterone (T) on the proliferation of MRC-5 cells, (2) to investigate the role of E and T on the viability of MRC-5 cells exposed to physiological and supraphysiological levels of LDL, and (3) to evaluate the morphological changes associated with E, T, and LDL, alone or in combination, on MRC-5 cells. Proliferation rates, biochemical marker analysis, and morphological evaluations were performed following standard laboratory procedures. The results of the investigation revealed that physiological and supraphysiological levels of LDL, alone, or in combination with E and T, induced significant changes in the functional and structural capacities of MRC-5 fibroblasts throughout the experimental phases (24, 48, and 72 hours) in comparison to the control groups. MRC-5 cells exposed to T and LDL resulted in increased proliferation rates and remarkable cellular damage. In contrast, E exposure induced decreased levels of MDA compared to T exposure. Results from this investigation suggest the development of collagen matrix post cardiovascular necrosis can be attributed to the presence of T. This response could be triggered directly at the fibroblast level or by altering the physiochemical characteristics of LDL. PMID- 12085601 TI - The effects of TCP fabricated with amino acid binders on the proliferation and viability of fibroblast cells. AB - Calcium phosphate (CaP) biomaterials were produced, by fusing individual crystals together at 1200 degrees C. This ceramic material has the ability to interact with and bond to the surrounding tissue. Therefore, ceramics can serve as scaffolds for tissue growth. The objective of this study was to determine if ceramics formulated with amino acid binders can enhance and support cell growth. A total of 147 tubes containing 1 x 10(5) MRC5 cells were divided into seven equal groups. Group I served as control, Groups II-VI were incubated with ceramics containing amino acid binders (ASP, CYS, HIS, LYS, or SER, respectively) and Group VII served as ceramic without binder control. Equal numbers of tubes were terminated from each group at 24, 48 and 72 hours. Cell numbers, morphology and markers of cell damage were determined at each time point. Results indicated: 1) Total cell numbers at 24 hours were increased in all experimental groups when compared with control. Increased levels of MDA were associated with control cells. 2). At 48 hours the total cell numbers in comparison to the control cells were increased in all groups with the exception of HIS. In addition to cell number, cellular adhesion was also increased in the experimental groups compared with the control. Significant increases in MDA levels were detected at 24 hours in the CYS treated cells. 3). Cellular adhesion in the experimental groups was increased over the control, but the total cell number was only increased in the ceramic material groups containing CYS and LYS. Cells exposed to ceramics with SER as a binder showed decreased cell numbers as well as increased MDA levels at this time point in comparison to control and other binders. Morphological evaluation of the cells showed amino acid binders with acidic side chains caused an increase in the presence of spindled shaped cells. Neutral side chains resulted in cells with elongated appearances, and basic side chains caused the cells to be less elongated. In conclusion, amino acid binders are able to induce the cells to adhere, proliferate and affect their morphological appearance as early as 24 hours. PMID- 12085602 TI - Investigation of the relationship between arch height and maximum pronation angle during running. AB - A three-dimensional motion analysis study is performed on 8 subjects running on a treadmill. Medical and lateral longitudinal arch height is measured from a weight bearing x-ray. A statistical analysis is performed to search for a relationship between arch height and maximum angle of pronation. The data analysis showed a significant relationship (sig. level = 0.0252) between the medial longitudinal arch height and the maximum eversion angle. The data analysis also showed a significant relationship between the lateral longitudinal arch height and the maximum eversion angle (sig. level = 0.0358). PMID- 12085603 TI - Understanding recumbent cycling: instrumentation design and biomechanical analysis. AB - The recumbent cycling position (RCP) is gaining popularity, with both recumbent ergometers and bicycles available as an alternative to the standard, upright cycling position (SCP). However, very little is known about the RCP, creating a situation where uniformed choices are being made. A system was developed for measuring and analyzing the forces placed on a bicycle pedal during operation of a stationary ergometer in either the RCP or SCP. Clipless pedals were modified so that forces normal and tangential to the surface of the pedal could be measured in the plane of the bicyclist. Additionally, the angle of the pedal was measured relative to the crank arm, as well as crank arm angle, so that pedal force orientations in a global reference were known. The pedals were modified in such a way that they were structurally and functionally similar to their original design. After obtaining university-approved informed consent, a single RCP with similar included angle between torso and lower extremities to the SCP was compared by having 19 recreational cyclists pedal in both positions (60 rpm, 250 W). Both pedal forces and lower-extremity kinematics were measured and an inverse dynamics analysis was performed. The energy transferred from the upper body to the lower extremities was significantly reduced in the RCP. Additionally, the amount of work done by knee flexion was significantly increased in the RCP while total work at the knee was not different. Due to these differences, depending on the situation, it may be more advantageous to select one cycling position over the other. PMID- 12085604 TI - Implantation of multiple intercostal nerve neuromas in lesioned spinal cord repair, regeneration and rehabilitation strategies. AB - Multiple intercostals nerve neuromas are surgically implanted in the lesioned spinal cord by bridging transaction lesions at T6 or T8 by removing one centimeter of spinal cord. In these experiments one to four intercostals nerves with origin proximal to the transection and the distal stumps, the neuroma was implanted distal to the transection. These procedures plus rehabilitation resulted in enhancing of regeneration of axons in the spinal cord lesion and dogs developed standing, stepping and reflex walking locomotion. Surgical resection of the implanted nerve resulted in paraplegia. Multiple nerve implants bridging the transaction of 34 mature female dogs. A pattern of neurological deficits developed when a sequence of resection of the implanted nerves were performed. If four nerves were implanted and one nerve resected usually no deficits were observed. When the second nerve was resected certain deficits were observed. When three nerves were resected some dogs became paraplegic, and some dogs showed increased sign of paraplegia but were not completely paralyzed and showed hind limb movements but no weight bearing. When four nerves were resected full paraplegia was observed. PMID- 12085605 TI - Mechanical characterization of a novel cell stimulating system (CSS) to apply dynamic, uniform and isotropic biaxial strains to cells in vitro. AB - Mechanical loading alters cellular responses. While in vitro mechanical stimulation is a powerful tool for exploration of mechanotransduction, very little has been published documenting techniques for validation of such devices. We have developed an in vitro experimental system that imposes well-defined temporal and spatial strain profiles using a pressure-actuated, tethered diaphragm substrate (Bioflex dishes). More importantly we have accurately characterized the strain and strain rate performance of this system and herein describe that methodology. The prototype CSS deflected cell substrates over cylindrical platens, producing dynamic biaxial strains. Dynamic studies at 1 Hz were conducted at 8.0, 9.0, 10.0 and 13.0 kPa peak transmural pressures for a total of 1000 loading cycles. To study the effects of frequency, experiments were also run at 0.5 and 1.5 Hz at 8 and 13 kPa. A series of 33 dots were placed collinearly in rings on the membrane. Dot motions were monitored via a CCD video camera and acquisition was performed using an 8-bit gray-scale video board and N1H Image software. Strain fields and rates were subsequently calculated using Mathematica software. Results confirmed that the strains were biaxially uniform over the frequencies and pressures examined: e.g., at 9.0 KPa, max radial & circumferential strain = 0.009 +/- 0.001. It was also shown that, as transmural pressure was increased, both membrane strains and strain rates increased; however biaxial strain isotropy was preserved. While we cannot measure out-of-plane deflections, video-based image analysis is a very useful technique for validation of dynamic planar biaxial strains in cell stimulation systems. PMID- 12085606 TI - Analysis of pin force distributions of halo orthoses. AB - The halo orthosis is a medical device used to immobilize the cervical spine when an injury to the region is sustained. This device consists of a halo vest, support rods, a ring, and four pins that contact the skull. A major problem experienced with the halo is pin loosening. In our previous research, we were able to model the halo orthosis as a mechanism and use kinematic principles to analyze the constraint characteristics. Using the Grubler/Kutzbach equation (a well accepted method of characterizing constraint systems), we were able to show that the halo orthosis is over-constrained which could be a possible source of pin loosening. We now present preliminary data that will show decreasing axial pin force and observed geometry changes that occur at each pin site. Biomechanical tests were performed on two cadaver skulls using a tensile loading machine that simulated the forces exerted by a patient. We were able to observe the changes in axial forces at each of the pins using load cells as cyclic transverse forces were applied. Our results show a force reduction over time at each pin site. We also discovered that the effects of creep are significant and potentially contribute to pin loosening. PMID- 12085607 TI - Intervertebral rotations as a function of rear impact loading. AB - Rear impact loading of the cervical spine results in a complicated biomechanical problem due to the complex geometry and viscoclastic material properties of this anatomy. Although a number of investigations have been performed to understand the biomechanics of rear impact, the dependence of segmental kinematics on cervical level and input velocity has not been clearly outlined. An experimental investigation was performed for this purpose using 10 isolated head-neck specimens. Segmental motions of the cervical spine were obtained for levels C2-C3 to C6-C7 at rear impact velocities of 2.1, 4.6, 6.6, and 9.3 km/h. Increases in segmental motion from baseline kinematics were compared for the three higher velocity tests. Results indicated greater increase in magnitude of segmental motion for levels C5-C6 and C6-C7 at higher input velocities than all other investigated levels. This finding helps to define the kinematics of the cervical spine leading to whiplash injury. PMID- 12085608 TI - Dynamic modeling of the neck muscles during horizontal head movement. AB - This paper presents modeling and simulation of superficial neck muscle movement in the horizontal plane (yaw). The parametric muscle model was constructed using Pro/Engineer 2000i Student Edition, Parametric Technologies Corp, and simulated using Pro/Mechanica. Pennation angles, force-tension, force-generation and rate of muscle activation data were obtained from anatomic and physiological studies. Saccadic eye movement models developed by G. Alexander Korentis and John Enderle also provided the basis for this model. PMID- 12085609 TI - Center of pressure measures to assess standing performance. AB - Functional neuromuscular stimulation (FNS) systems have provided individuals with spinal cord injury (SCI) varying levels of standing capability. Our long-term goal is to enhance an FNS user's ability to perform activities of daily living by providing sufficient standing stability. However, control of standing posture with complete SCI is compromised by a lack of sensory feedback from receptors in the feet. The goals of this study were: a) to document a variety of center of pressure (COP) measures during standing, b) to compare able-bodied and FNS standing, and c) to relate various measures to overall quality of standing performance. Two subjects with complete SCI that stand with FNS (one with a surface system and one with an implanted stimulation system) and five able-bodied subjects stood quietly on force platforms. Reduced hand support forces were coincident with anterior/posterior (A/P) mean COPs that were further posterior in the base of support and medial/lateral (M/L) mean COPs that were more centered between the feet. In addition, M/L COP excursions, maximum A/P COP velocities, and maximum M/L COP velocities were all significantly greater for FNS users than for able-bodied standers. Furthermore, maximum A/P COP velocities and mean A/P COP frequencies were significantly greater during FNS standing trails that required increased support from the hands. COP measures such as these may be useful tools to assess standing performance and to determine strategies for adjusting posture that can reduce an FNS user's reliance on upper extremity support while maintaining stable balance. PMID- 12085610 TI - A new method of tightening halo orthosis pins that reduces initial pin force variation. AB - The halo orthosis is a medical device used to immobilize the cervical spine when an injury to the region is sustained. This device consists of a halo vest, support rods, a ring, and four pins that contact the skull. The halo is applied to the patient by securing the halo ring to the skull with four pointed pins that are threaded into a halo ring. In an effort to produce a desired initial force in each pin, a specified torque (typically 0.9 N-m) is applied to the pin. One problem associated with the existing method is that it produces a wide range of initial pin forces despite a constant applied torque. The existing method is particularly sensitive to the presence of medical agents (e.g. antibacterial ointments or hydrogen peroxide) that may change frictional characteristics at the pin site or at the threaded hole in the halo ring. Variability in initial forces from an applied torque of 0.9 N-m (8 in-lbf) have been reported in the literature. These forces vary from 620 N (140 lbf) when hydrogen peroxide is applied to the threads and pin site up to 980 N (220 lbf) when antibacterial ointment is applied to the threads and pin site. We present a new method of tightening halo pins that reduces the variability of the initial pin force. Using the new method, a known force is applied to the pin to provide a preload. Once the appropriate preload is applied, the halo pin is secured in place relative to the halo ring and the applied force is removed leaving the pin in the final configuration. We compare the existing method and the new proposed method by examining a single pin site. The pin force data display less sensitivity to lubricants using the new direct loading technique. PMID- 12085611 TI - Abstracting data warehousing issues in scientific research. AB - This paper presents the design and implementation of the Idaho Biomedical Data Management System (IBDMS). This system preprocesses biomedical data from the IMPROVE (Improving Control of Patient Status in Critical Care) library via an Open Database Connectivity (ODBC) connection. The ODBC connection allows for local and remote simulations to access filtered, joined, and sorted data using the Structured Query Language (SQL). The tool is capable of providing an overview of available data in addition to user defined data subset for verification of models of the human respiratory system. PMID- 12085612 TI - A comparison of two methods for choosing the regularization parameter for the inverse problem of electrocardiography. AB - We have previously compared both generalized eigensystem (GES) and Tikhonov regularization methods for estimating epicardial potentials from measured body surface potentials. Both of these methods require the choice of a regularization parameter. In this study we compare two methods for choosing this parameter: the Composite Residual Error and Smoothing Operator (CRESO) method, and a new Zero Crossing (ZC) method. We compared both CRESO and ZC methods for zero and first order GES and Tikhonov regularization methods on a swine model and found that the ZC method sometimes produces smaller errors, but only for small noise levels. PMID- 12085613 TI - Thresholding reflux episodes in impedance measurements using a neuro-fuzzy system. AB - Multiple intraluminal impedance measurements (MII) provides impedance signals which reflect the changes in the electrical conductivity inside the gastrointestinal tract (GIT). To determine a reflux episode from MII signals, wavelet transform is used to detect the edges of the impedance, then thresholding impedance at certain level. A predictor for the value of the threshold is constructed by training a fuzzy system using a number of features. During training process the predictor clusters the features into classes of similar feature values. The trained system is used to predict the threshold applied on the distal channel. Subsequently, a combination of wavelet transform and fuzzy logic is used to classify the episodes. In this paper we present the results of one stage of the system that predicts the threshold applied on the impedance value. PMID- 12085614 TI - Time frequency analysis of the cardiovascular response during the intracarotid amobarbital test. AB - Recently, time-frequency analysis has become very popular for examining non stationary time series and for researching fast changing phenomena. We used the smoothed-pseudo Wigner-Ville distribution to model the underlying dynamic autonomic nervous system changes during the intracarotid sodium amobarbital (ISA) or so-called Wada test. The Wada test involves injecting sodium amobarbitol into the internal carotid artery that results in inactivation of cerebral structures supplied by the ipsilateral anterior and middle cerebral arteries. Electrocardiogram (EKG) data were recorded during the entire procedure and the 10 minutes prior to each injection were used as baseline values for that hemisphere. Interbeat-interval time series were created from these data for a 13 year old patient and were examined using a smoothed-pseudo Wigner-Ville distribution. Inspection of these data indicated that sodium amobarbitol injection to either side produced decreased power in the vagally mediated high frequency band (.14 .40 Hz). Importantly, this decrease was greater when the right hemisphere was inactivated as compared to the left. These results are consistent with the known lateralized innervation of the heart such that right-sided autonomic inputs have greater influence on cardiac chronotropy. The present results also revealed the very rapid changes in autonomic control that characterized the inactivation and subsequent recovery of the cerebral hemispheres. These findings confirm the utility of time-frequency analysis in the investigation of cardiac time series. PMID- 12085615 TI - Medical technology management in U.S. hospitals. AB - Medical technologies move from research and development through manufacturing and marketing into the healthcare delivery system. Within the healthcare delivery system, hospitals rely heavily on medical technologies (and the medical devices they enable) to provide diagnosis, treatment, and monitoring in patient care. Managing these devices from acquisition through application in patient care is a formidable task. Hospitals must act to maximize the benefits of medical devices while minimizing adverse side effects. They must do so within a highly regulated and cost-constrained environment. This paper describes the challenges hospitals face and the strategies they employ in their efforts to achieve cost-effective medical technology management. The role of clinical engineering is discussed. PMID- 12085616 TI - First-order system least squares for elastohydrodynamics with application to flow in compliant blood vessels. AB - Mathematical modeling of compliant blood vessels generally involves the Navier Stokes equations on the evolving fluid domain and constitutive structural equations on the tissue domain. Coupling these systems while accounting for the changing shape of the fluid domain is a major challenge in numerical simulation. Many techniques have been developed to model compliant vessels, but all suffer from disproportionate increase in computational cost as problem complexity increases (i.e., larger domains, more dimensions, and more variables.). Even the best standard methods result in computational cost that typically grows quadratically with the degrees of freedom. Using a least-squares formulation of the problem, elliptic grid generation for the changing fluid domain, and an algebraic multigrid solver for the linear system can overcome many shortcomings of standard techniques. Most notably, the computational cost of solving the problem increases linearly with the degrees of freedom and the associated functional provides an a posteriori error measurement. Least squares represents a systematic approach for formulating the original problem so that the numerical process becomes straightforward and optimal, and it avoids restrictions that often limit other methods. Results are presented for a two-dimensional test problem consisting of a Newtonian fluid with properties of blood in a linear elastic vessel with properties of smooth muscle. PMID- 12085618 TI - A closed-loop controller for mechanical ventilation of patients with ARDS. AB - Mechanical ventilators are routinely used to care for patients who cannot adequately breath on their own. Management of mechanical ventilation often involves a careful watch of the patient's arterial blood-oxygen tension and requires frequent adjustment of ventilation parameters to optimize the therapy. This situation lends itself as a candidate for closed-loop control. This report describes a closed-loop control system based on well-established protocols to systematically maintain appropriate levels of positive end-expiratory pressure (PEEP) and inspired oxygen (FiO2) in patients with Adult Respiratory Distress Syndrome (ARDS). The closed-loop control system consists of an in-dwelling arterial oxygenation (PaO2) sensor (Pfizer Continucath), coupled to a Macintosh computer that continuously controls FiO2 and PEEP settings on a Hamilton Amadeus ventilator. The implemented protocols provide continuous closed-loop control of oxygenation and a balance between patient need and minimal therapy. The controller is based on a traditional proportional-integral-derivative (PID) approach. The idea is to control, or maintain, the patient's PaO2 level at a target value determined, or set, by the patient's physician. The controller also features non-linear and adaptive characteristics that allow the system to respond more aggressively to "threatening" levels of PaO2. Another benefit of the control system is the ability to display, monitor, record and store all system parameters, settings, and control variables for future analysis and study. The system was extensively tested in the laboratory and in animal trials prior to use on human subjects. The results of a small clinical trial indicated that the system maintained control of the patient's therapy nearly 84% of the time. During the remainder of this time, the controller was interrupted primarily for suctioning, PaO2 sensor calibration or replacement. The response of the closed loop controller was found to be appropriate, reliable and safe in patients with ARDS. PMID- 12085617 TI - Impacts for medicine of global monitoring. AB - In his 1998 Turing Award speech, Jim Gray described a number of research goals including those of building what he referred to as a Personal Memex and a World Memex. The Personal Memex is a system for recording everything one saw, heard, or read, while the World Memex is a system to contain all professionally produced information. In this paper we discuss the consequences to medicine of an additional type of monitoring, that of movement and position via GPS devices. The paper argues that such devices will be incorporated into hand-helds, telephones, and wristwatches, and that a World Memex will (with appropriate permissions) monitor and record all personal movement. The motivation for such a development is the many uses to which the system can be put. The paper restricts itself to discussing those uses that apply to physical safety and medical research studies. Examples relating to safety include the detection of, and notification of emergency authorities about, accidents involving unusual motion, such as occur e.g. in car accidents or when people fall off ladders. Examples of medical research studies include those that involve the effects of exercise, or exposure to different environmental conditions. Precise quantitative statistics can be gathered, providing answers to such questions as what the optimum amounts of exercise are for various health-related conditions. The paper discusses these among other such applications. PMID- 12085619 TI - Histomorphometric analysis: comparative study of manual and digital image analysis using reproductive organ as a model. AB - A preliminary analysis of the histological features of testes was carried out on vertical serial sections stained with hematoxylin and eosin techniques. The total spermatogonia per view and areas of seminiferous tubules by means of digitizing the tissue followed by morphometric analysis using software run on a personal computer. The data generated was then compared with traditional manual determinations made by using the ocular scale as well as the x and y coordinated on the light microscope. The data obtained showed that the modified Image Pro system for digitizing and data analysis allowed for high-resolution acquisition of the images as well as relatively high sensitivity of image integration. The results also show more accuracy and repeatability of sampling and a decrease in sampling error between repeated measures compared with the conventional manual measurements. Overall, the use of modified Image Pro digital technology will offer a more accurate and quantitative method to perform routine histomorphometric analysis, as well as allow for repeated measurements of the same sample from several reviewers in a blinded fashion with substantially less error. PMID- 12085620 TI - Multiple spatially distributed stimulators and timing programs for entrainment of activation during ventricular fibrillation. AB - Activation sequences during ventricular fibrillation (VF) display complex pattern and fast rate. Recent evidence suggests that even during VF an excitable gap may exist. Existence of excitable gap lead us to hypothesize that it should be possible to entrain activation patterns during VF by using spatially distributed and temporally phased pacing strength stimuli. We describe here the electronics hardware and software that were developed to test our hypothesis. Eight biphasic stimulators were designed and fabricated, each addressable via a TTL input and thus independently triggered. To minimize electrical interference from stimulus pulses the stimulators were optically isolated. A program written in C was used to deliver TTL inputs to time the sequence of stimulation. The parameters that could be varied were, pulse intensity, polarity, and duration, inter pulse interval and activation pattern cycle length. Restitution's of action potential duration, conduction velocity, and complex activation patterns make the timing of stimulators complex. To aid in optimal timing of these stimulators, we used a Luo Rudy ionic model of cellular activation to simulate VF in a matrix of 400 x 400 cells. Entrainment of activation was verified using animated displays of activation sequences. Using results of simulation we verified the function of our stimulators experimentally using electrically induced VF in canines and by using electrograms recorded from 121-electrode patch. Our results show that the developed hardware and software can be used to deliver distributed stimuli in a flexible and effective pattern, which may aid in development of approaches for treatment of VF. PMID- 12085621 TI - Assessment of ventilatory sensitivity to carbon dioxide changes during orthostasis. AB - Investigators have anecdotally reported changes in respiratory pattern preceding symptoms of orthostatic intolerance for several years. Evidence from recent studies support these observations, and show that alterations in respiration often precede pre-syncope during orthostasis. These observations suggest the possibility that changes in interaction between the chemo and baro-reflex control systems may produce phasic or dynamic changes in respiratory and hemodynamic parameters such that these changes are important in orthostasis intolerance. Our objective in the present study was to develop a method to obtain a quantitative index of ventilatory sensitivity to changes in inspired during orthostasis. Based on an approach previously developed by one of the authors (EB), we used, during supine and 70 degrees head up tilt, pseudo-random binary changes in inspired CO2 to quantify ventilatory sensitivity to CO2 disturbances. A pneumotach was used to measure airflow. From these recordings, we used a prediction error based systems identification algorithm to quantify ventilatory impulse response to CO2 stimulus. Results from 7 subjects showed that the integrated ventilatory response (area under impulse response curve) to CO2 was larger during tilt than that during supine (mean [std] 5.3 [2.6] Vs 3.0 [1.6] ml/min/0.01LCO2). These results suggest that the changes in ventilation due to disturbances in CO2 are likely to be larger during orthostasis than those during supine. Such an elevated response would indicate that the role of chemo-reflex control of breathing in cardiovascular instability culminating in orthostatic intolerance is likely to be important and should be further investigated. PMID- 12085622 TI - Regression analysis for vortex ring characteristics during left ventricular filling. AB - A new technique to determine ventricular function is being developed from non invasive measurements of centerline mitral flow, using color M-mode (CMM) ultrasound. Based on the hypothesis that vortex rings accompanying mitral flow are significant indicators of ventricular filling, we present a technique to determine characteristics of vortex rings created by mitral inflow, including diameter, position and circulation from the CMM derived centerline velocity data. The rings are modeled as vortex filament loops, which induce axial flow. A quasi Newtonian regression was performed on the velocity data to determine model ring characteristics. Performance of the technique was determined using a synthetic data set with known vortex ring parameters. The algorithm is shown to be insensitive to initial estimates for ring parameters. However, noise in the data can lead to inaccuracy in the derived ring parameters. Further work to increase the accuracy of the regression algorithm and decrease effect of data uncertainty is ongoing. PMID- 12085623 TI - Development of a learning module using a virtual environment to demonstrate EMG and telerobotic control principles. AB - A prototype system was developed for use as a teaching tool, allowing students to link EMG monitoring, data smoothing, robotic control, and neural network training within a rapid prototyping virtual environment (VE). The VE software allowed for the rapid development of scenarios and, when linked with EMG data input to a neural network, allowed the user to control an artificial world containing a virtual arm. Student teams then attempted to control the arm in the VE while performing the tasks by use of a neural network system they had specifically developed and trained using their own EMG signals. The results from their system were then translated into a form that enabled the control of a real robot. Students enjoyed the challenge and uniqueness of the module, and were enthusiastic about extending the concept to other areas of interest. PMID- 12085624 TI - Evaluation of the web-based "home helper" support system using wireless Internet mobile phones. AB - In Japan, Home Helpers provide the home welfare and care services such as cooking, bathing, washing, cleaning, shopping, etc. Last year, we developed the web-based Home Helper support system using wireless Internet mobile phones for improving scheduling and record keeping efficiency and for eliminating unnecessary travel. We have evaluated by questionnaire whether the system can be easily operated. All in all, the system has performed satisfactorily and is in functional use daily. PMID- 12085625 TI - Reverse flow in compliant vessels and its implications for the Fontan procedure: numerical studies. AB - The Total Cavopulmonary Connection (TCPC), a variant of the Fontan operation used for palliative cardiovascular repair of patients with single ventricle physiology, creates a passive system of blood flow into the pulmonary circulation for which energy efficiency may be critical to long term outcome. Clinical studies have shown that reverse flow in the TCPC is an indication of poor clinical status in these patients. Using numerical simulations, we demonstrate that reverse flow leads to increased energy losses in compliant vessels. Such an effect can potentially set off a series of spiraling negative events with decreased ventricular function leading to reverse flow, which causes decreased energy efficiency, which in turn leads to worsening function, and so forth, thereby suggesting one cause of progressive heart failure in this patient group. PMID- 12085626 TI - Mathematical model of a variable aperture mitral valve. AB - Previous models of pressure and flow waveforms during diastolic ventricular filling by the Waite et al. [2] described mitral valve behavior by a constant effective valve area and a constant valve impedance. A model to describe the opening process of the mitral valve was developed. The model is based on second order dynamics and Bernoulli's equation. The earlier lumped variable model for diastolic filling was modified to reflect a variable mitral valve aperture. The combined model was calibrated using pressure and flow waveforms from porcine experiments and artificial transmitral pressure gradient waveforms. A comparison between the results of the combined model and the earlier fixed aperture model shows better agreement of experimental and simulated transmitral flow waveform in the combined model. PMID- 12085627 TI - Investigation of deformability, viscosity, and aggregation of mPEG-modified erythrocytes. AB - Several techniques for the development of a universal blood type are under investigation in order to address the vast shortage of blood required during times of crisis and for those receiving chronic transfusions. Recently, the covalent attachment of methoxypolyethylene glycol (mPEG) has received attention as a means to conceal the major and minor blood antigens that can invoke an immune response. We have utilized a simple method to covalently bind mPEG to the surface of red blood cells and monitored the rheological properties including cellular deformability and suspension viscosity as a function of shear rate and aggregation. In order to simulate the deformability of mPEG-modified erythrocytes through small blood vessels, we have studied the flow of cells through a nickel mesh membrane with a pore size of 5.8 microns. Using a gravity-based nickel-mesh filtration device, dilute solutions of red blood cells were filtered through a membrane. The pressure drop was monitored with time. We have examined the filterability of whole blood, washed erythrocytes, mPEG-altered erythrocytes, and mixtures of altered erythrocytes with whole blood. Results from this apparatus have suggested that there is a significant decrease in deformability using mPEG modified erythrocytes compared to unaltered red blood cells at identical concentrations. Increasing the ratio of modified cells to unmodified cells resulted in decreased deformability of the suspension. Viscosity measurements of mPEG-modified red cells have shown a shear-thinning property, while results of aggregation measurements indicate reduced aggregation compared to whole blood. PMID- 12085628 TI - Use of an infrared temperature monitoring system to determine optimal temperature for laser-solder tissue repair. AB - Thermal damage of tissue is a major concern for all laser tissue repair techniques where the resulting strength of a repair is sensitive to slight alterations in tissue temperature as well as changes in the duration of exposure of the tissue to the laser beam. Too low of a temperature will prevent proper bonding between the surfaces while prolonged exposure of the tissue to the laser beam results in collateral thermal damage and decreased flexibility and strength of the repair. Temperature feedback systems that monitor the surface temperature of the repair site and adjust the laser irradiance accordingly increase the success rate of the technique. Knowledge of an optimal temperature for tissue soldering will also increase the reliability of the technique. The choice of solder material has been another challenge to the reproducibility of strong repairs. The emerging use of solder-doped polymer membranes as surgical adhesives offers numerous advantages over more traditional liquid and solid solders. Poly (L-lactic-co-glycolic acid) (PLGA), when used as a polymer scaffold, is porous enough to absorb serum albumin and can also be doped with various hemostatic and thrombogenic agents to aid tissue healing. An in vitro study was performed to correlate tissue temperature with the tensile strength of repairs formed using the solder-doped polymer membranes. Previous studies by our group indicate that a solder/tissue interface temperature of 65 degrees C is optimal. Using this parameter as a bench mark, laser irradiance was varied and the solder surface and solder/tissue interface temperatures were monitored by an IR temperature monitoring system, designed by the researchers, and a type K thermocouple, respectively. PMID- 12085629 TI - Solubility studies of albumin protein solders used for laser-assisted tissue repair. AB - Albumin protein solders used for light-activated tissue repair are soluble in physiological fluids prior to laser irradiation. Inevitably, some of the material tends to run away before it can be bonded to the tissue. In addition, blood dilution alters the mechanical properties of the solder. Consequently, the strength of the repair is compromised resulting in poor reproducibility and reliability of the repair technique. The emerging use of solder-doped polymer membranes offers a potential solution to this problem. Predenaturation of the solder in a hot water bath also holds promise as a method for reducing initial solder solubility. The purpose of this study was to determine if polymer reinforcement and/or predenaturation could reduce the solubility of albumin protein solders, and thus, the ultimate breaking strength of incisions repaired by means of laser soldering techniques. A Bradford protein assay was utilized to measure the solubility of the protein solders prior to and after thermal denaturation. The results of this comparison showed that doping of the solder in a polymer membrane and predenaturation of the solders at 75 degrees C were advantageous for improving their handling characteristics. Alteration of the mechanical properties of the solders prior to laser treatment was also prevented, thus improving the reproducibility and reliability of the repairs. PMID- 12085631 TI - Optimization of laser-solder repair technique for possible application in strabismus surgeries. AB - Strabismus is the lack of binocular vision due to an inability to control one of the eye muscles. Corrective surgery is the most common recourse and consists of adjusting and reattaching the extraocular muscle to the sclera. In approximately 10% of cases involving re-insertment of the extraocular muscle via suture techniques, the needle is inserted too deeply into the eye resulting in perforation of the retina. Fibrin glues and cyanoacrylates have been substituted with unsatisfactory mechanical results. The goal of this study was to maximize the tensile strength of rabbit extraocular muscles repaired using a laser-solder technique developed by McNally et al., Biodegradable polymer membranes of controlled porosity were fabricated with poly(L-lactic-co-glycolic acid) (PLGA) and salt particles using a solvent-casting and particulate-leaching technique. The porous membranes were doped with protein solder composed of 25% and 50% (w/v) serum albumin and 0.5 mg/ml indocyanine green (ICG) dye mixed in deionized water. In vitro tissue specimens were repaired using the solder-doped polymer membranes in conjunction with an 805 nm diode laser. The tensile strength was tested on an MTS machine and results were analyzed with the Student's T-test. PMID- 12085630 TI - Towards tissue engineering of a composite aortic valve. AB - A tissue-engineered valve needs to incorporate the complex microstructure of the native aortic valve if it is to be as durable as existing bioprosthetic valves. Native aortic valve cusps contain large collagen fiber bundles surrounded by tubes of elastin, linked together by elastin sheets and struts. They also contain glycosaminoglycans (GAGs) that bind water and give the valve cusp a gelatinous consistency. Our approach to tissue engineering the aortic valve is to fabricate the cusp from the building blocks described above. We have developed collagen fiber bundles using the principle of directed collagen gel shrinkage, a GAG matrix by crosslinking high molecular weight hyaluronan with divinyl sulfone, and elastin sheets and tubes by culturing neonatal aortic fibroblasts on the appropriate substrates. To make collagen fiber bundles, cells are mixed with solubilized fibrillar collagen and cast into silicon rubber wells fitted with microporous holders to entrap the gel and hold it in place. As the cells interact with the collagen fibrils, they contract and compact the gel. Since the gel is constrained in the longitudinal direction, it can contract only laterally, forming well-aligned, strong (> 1 MPa) collagen fiber bundles. Elastin sheaths from naturally around the collagen fiber bundles and atop the hyaluronan gel when the neonatal aortic fibroblasts are cultured for more than 4 weeks. The individual building blocks of the aortic valve cusp, designed and fabricated according to patterns dictated by the microstructure of the native aortic valve, will be stacked together to make the final composite, tissue-engineered aortic valve cusp. PMID- 12085632 TI - Blue light dermal imaging of human skin. AB - While the number of occurrences of skin cancer is constantly increasing, the medical industry relies on the same slow and invasive method of detection. Image processing techniques could prove to be a useful diagnostic tool. With a blue light laser, an optical system, and imaging techniques, a noninvasive detection method that produces immediate results could be developed. This paper analyzes the feasibility of developing such a detection device. The evaluation included the safety and effectiveness of using a blue laser for dermal imaging. The data came from research and a preliminary laboratory experiment. The analysis showed that this laser-based method of skin cancer detection is safe, effective, and continued research should be performed in this area. PMID- 12085633 TI - Image processing and 3D reconstruction of serial section micrographs from Musca Domestica's biological cells responsible for visual processing. AB - The ability to visualize and understand three-dimensional objects from two dimensional cross-section or slice images is difficult, even if the observer has a general concept of the object of interest. The focus of this research is to apply image-processing methods to two-dimensional cross-section electron transmission micrographs of the biological cells of the Musca Domestica's, or household fly's visual system in an effort to better understand the cells responsible for processing visual information. The application of knowledge gained from biological systems is know as biomimetics. The first task will be to construct a useful three-dimensional data set from two-dimensional micrographs provided by the U.S. Air Force Academy in Colorado Springs, Colorado. The data set will be constructed by aligning these images in an edge-to-edge fashion to form a layer. Once each layer is reconstructed, the layers will be stacked and registered to form the third dimension of the data set. This task is complicated by the fact that translation, rotation and scaling mismatches exist in the images. The second task will be to segment and label the biological cells of interest. Computerized segmentation has not yet proved successful, resulting in a manual or "brain-powered" approaches being used at many institutions. By using and modifying current computer image-processing techniques, advances leading to a semi-automated segmentation process may result. Finally, the segmented data must be formatted for use with existing software to render and view the cell(s) of interest. A "marching cubes" surface-rendering algorithm is often implemented in current visualization software, along with routines to view, rotate and scale the resulting surfaces in real time. The result of viewing and manipulating the biological data set will be an increased understanding of the processes of the fly's visual system. Other researchers will use the knowledge gained from the three-dimensional renderings of the cells to further develop an analog vision system based on the fly's compound eye. Much of this research is funded by the Navy Air Warfare Center in an effort to design an analog visual system with real time target identification and tracking capabilities. PMID- 12085634 TI - A fuzzy-based texture analysis for tissue characterization of diffused liver diseases on B-scan images. AB - The specific texture on B-scan images is believed to be related to both ultrasound machine characteristics and tissue properties, i.e., the pathological states of the soft tissue. Therefore, for classification, features can be extracted with the use of image texture analysis techniques. In this paper a novel fuzzy approach for texture characterization is used for classification of normal liver and diffused liver diseases, here fatty liver, liver cirrhosis, and hepatitis are emphasized. The texture analysis techniques are diversified by the existence of several approaches. We propose fuzzy features for the analysis of the texture image. For this, a membership function is constructed to represent the effect of the neighboring pixels on the current pixel in a window. Using these membership function values, we find a feature by weighted average method for the current pixel. This is repeated for all pixels in the window treating each time one pixel as the current pixel. Using these fuzzy based features, we derive three descriptors: maximum, entropy, and energy as used in co-occurrence method, for each window. PMID- 12085635 TI - Examining a CT scan of an abdominal aortic aneurysm. Part 1: Measuring the thrombus to aneurysm ratio using MatLab. AB - Abdominal Aortic Aneurysms are a relatively common medical condition that occurs when the wall of the aorta expands to an above average diameter somewhere in the abdominal region of the body. Since an abdominal aortic aneurysm has a good chance of rupturing, which usually results in death, it must be detected and treated before rupture occurs. The current method of determining whether surgery is necessary depends on factors such as size and expansion rate of the aneurysm as well as the age and health of the patient. A proposed hypothesis is that the ratio of thrombus area to overall aneurysm area may also aid in determining whether surgery is necessary because it correlates to whether the aneurysm is stable or expanding. Currently the sizes of the aneurysm, thrombus and lumen are determined by manually measuring the diameter of the overall aneurysm and the diameter of the lumen, then employing these in the area formula for a circle to determine their respective sizes. The thrombus area is calculated by subtracting the area of the lumen from the area of the overall aneurysm. Since the aorta may not be perfectly circular in shape and this method is time consuming and error prone, an improved method for finding these areas is needed. This paper discusses a preliminary study of using MatLab as a tool to implement an algorithm that better approximates the area of the thrombus and overall aneurysm. PMID- 12085636 TI - Examining a CT scan of an abdominal aortic aneurysm. Part II: Locating the aneurysm using the Hough transform. AB - When analyzing CT scan images for the diagnosis of an Abdominal Aortic Aneurysms the aorta must be located and cropped from the whole image. Given that there are so many images to be analyzed, it is wise to automate this cropping process so further image-processing techniques can be applied. By using the Hough transform the process of finding and cropping the aorta from the image in a consistent method is important. The Hough transform lends itself to this process quite nicely. This paper focuses on implementing the Hough transform in C++ using Microsoft Windows. PMID- 12085637 TI - Microarray image spot segmentation using the method of projections. AB - Microarray technology is increasingly used as a means of high throughput analysis of human, non-human and plant genomes. Manual methods of array production using this technology lead to many inherent problems in the microarray image produced. The density of the spots in the images produced is very high, such that neighboring spots can overlap. The image background is often not uniform, containing noise that is often difficult to distinguish from actual spots. In this research, a projections-based approach is investigated for spot segmentation in paired radio probe microarray images. An important aspect of spot segmentation is the capability to perform corresponding spot-to-spot comparisons between the paired images. Experimental results are presented for spot segmentation in isolated and paired microarray images. PMID- 12085638 TI - Detection of dysplasia with near real time confocal microscopy. AB - The use of high resolution, in vivo confocal imaging may offer a clinical tool to detect early neoplasia and reduce the incidence and mortality of cancer. Our laboratory is currently examining the feasibility of using confocal microscopy for non-invasive diagnosis of dysplasia and early carcinoma in epithelial tissue. We are performing a series of ex vivo studies investigating the optical properties of normal and abnormal biopsies to quantify the diagnostic capability of this technology to discriminate between normal and pre-cancerous tissue. These studies use a near real time reflectance confocal microscope to acquire images at various depths throughout the epithelium. To date, we have completed a twenty five patient study of cervical biopsies and have acquired images from six patients of an approved twenty-two patient study in the oral cavity. The cervical study has shown a distinct difference between normal and dysplastic tissue which can be used diagnostically, while initial results from the oral cavity are promising even with increased keratin scattering. In conclusion, our examination of normal and precancerous biopsies has demonstrated the confocal microscope's ability to image sub-cellular morphology at a resolution making accurate diagnosis possible and supporting this technique's potential for in vivo assessment of dysplasia. PMID- 12085639 TI - Designing an Internet-based collaboratory for biomedical research. AB - Several recent grants from the National Institutes of Health to the Universities of Wyoming, Idaho, and Montana have created a unique opportunity for collaboration in biomedical research among the three schools, as well as the community colleges in the region. NIH Center of Biomedical Research Excellence (COBRE) programs at Wyoming have been established to study the biological effect of nitric oxide and to investigate stressors that can contribute to the progression of cardiovascular disease. Funding from these and related grants have significantly upgraded Wyoming bioimaging and microscopy facilities, as well as provided support for faculty and students in a variety of research disciplines. In order to enhance these research efforts, the Center for Rural Health Research and Education at the University of Wyoming is spearheading an effort to create an Internet-based system for sharing data and research resources among the involved sites. This paper describes how such a "collaboratory" could be designed, using techniques developed for distributed research and development in the computer industry. The system, as envisioned, will support remote data acquisition, management, and visualization, while providing security in the form of authorization and authentication of users and virtual private networking for data transmitted between nodes of the network. PMID- 12085640 TI - Salvaging legacy data: mapping an obsolete medical nomenclature to a modern one. AB - The Veterinary Medical Database (VMDB) is a repository containing abstracts of over six million case records from 24 veterinary colleges throughout the U.S. and Canada. These case record abstracts, spanning almost 40 years, represent a valuable resource for outcomes analysis and hypothesis generation. Database records are currently encoded using the Standard Nomenclature of Veterinary Diseases and Operations (SNVDO), a precoordinated, hierarchical coding system. SNVDO has not been updated since 1977 and is outdated and inadequate to express the current state of medical knowledge. We undertook to manually map a subset of the SNVDO codes to a modern medical nomenclature, SNOMED-RT (Version 1.0), and to evaluate the quality of the resultant mappings and the acceptability of the mapping method used. We found that the distribution of frequency of use of the SNVDO codes in the VMDB records is highly skewed, with a small number of codes accounting for a large percentage of the records. We targeted our mapping efforts on that subset of codes. We found that our targeted manual mapping of the SNVDO codes to SNOMED-RT codes was feasible and produced good quality results, based on separate evaluations performed by two domain experts. However, a significant proportion of the SNVDO codes could not be mapped to a single SNOMED-RT concept, necessitating construction of multiple-code post-coordinated terms. Additionally, this manual mapping was very labor-intensive. PMID- 12085642 TI - A neural network characterisation of electromyography. Part I. AB - In this paper we seek to examine the extent to which motor unit firing sequences can be related to EMG signal sequences leading to the characterisation of tasks from their associated EMG temporal patterns. Arising from this, the work described here is concerned with the development of a computer model to simulate the muscular activity during elbow flexion. To achieve this we start by expanding upon models of the central nervous system (CNS) and the process of trajectory formation coupled with the use of a Neural Network to predict muscular activation, based on prerecorded EMG activation patterns for known tasks. In this way we can generate a pattern of muscular activation from physiological and kinematic data required to perform a task sequence. The EMG data, used in the networks training, has been taken from males aged between 18 to 26 years all without any known disabilities or injuries to the elbow joint. This paper reports on data processing and event detection used in this experiment and suggests further work in this field. PMID- 12085641 TI - Pifithrin-alpha (PFT-alpha) caused differential protection of rat liver cells and HepG2 cell line in response to the selective cytotoxicity of arsenic and cadmium. AB - In response to genotoxic agents, normal cells are instructed by p53 to either perform DNA repair or to commit suicide. Since chemo and/or radiotherapy damage both normal and cancerous cells, the use of PFT-alpha, a reversible inhibitor of down stream function of p53, was suggested as a temporary inhibitor of p53 induced cell damage. Our objective therefore, was (1) to assess the inherent response of HepG2 and rat liver cells to the effects of arsenic and cadmium and (2) to evaluate the role of PFT-alpha in the differential protection of rat liver and HepG2 cells. Following cellular growth to 90% confluency, exposure to cytotoxic agents in presence of PFT-alpha (10 ppm) or its absence was performed. Cell survival was detected fluorometrically using fluorescein diacetate (FDA) and an Ascent Fluoroskan. Toxicity index (LC50) was calculated from percent survival using regression analysis. Results showed an average of 46 fold inherent resistance of rat liver cells to arsenic in comparison to HepG2 cells (LC50 range of 573.15-670 vs. 13.4-13.7 ppm respectively). An average of 8 fold inherent resistance was also attributed to rat liver cells in response to cadmium (LC50 range of 57.72-58.1 vs. 6.99-7.35 ppm respectively). PFT-alpha did not show significant difference in protecting HepG2 cells against cadmium or arsenic. In contrast, there was significant difference in the protection of rat liver cells upon exposure to arsenic. We conclude that Pifithrin-alpha exhibits protection to normal cells, which can play an important role in cancer chemotherapy. PMID- 12085643 TI - Quantifying power output during cycling through measuring strain energy in a bicycle frame. AB - Quantifying power output is becoming more popular in training and competition for cycling. This popularity comes from the fact that power is a direct means of measuring intensity, which is considered by many to be the most important variable in a successful training program. Although other methods are available for measuring power output, there is no frame that is available that uses strain to quantify power output. Using a frame may prove superior to currently available products. Additionally, strain may prove useful for cycling technique analysis, another critical factor for success. In this study, a cyclocross bicycle frame was instrumented with six strain gages to determine if strain energy in the frame correlates with power output. For pilot data, an 85 kg subject rode the bike on rollers with a fork stand at 250 watts and a cadence of 90 rpm. Although more data needs to be collected and analyzed to find a correlation with power output, initial pilot data shows that the strain in the frame is cyclic in nature when pedaling and has a corresponding frequency with the pedaling cadence. It is hypothesized that the strain in the frame will have a high correlation with power output. While this application is specific to cycling, correlating strain to power output may prove useful in other applications. PMID- 12085644 TI - Accuracy of EEG source reconstruction in the presence of brain lesions: modelling errors and surface electrodes' placement. AB - Source localization techniques based on electroencephalography (EEG) use scalp potential data to infer the location of brain neural activity. A volume conductor model describing the electrical properties of the human head is needed. Lesions have conductivity considerably different from that of normal brain and should be included in the head model because the differences between the actual head and the model can cause source reconstruction errors. We performed a simulation study investigating EEG dipole source reconstruction errors, caused by brain lesions neglecting, using different measurement montages. The scalp was sampled by 64 electrodes (simulating clinical practice) and by 128 electrodes (extended configuration). The human head was represented by an eccentric-spheres model in which a modifiable eccentric bubble approximated various brain lesions. We analyzed 64 pathological situations. Results showed that neglecting brain lesions in source reconstruction procedures could cause large source localization errors which depended on source location and orientation, and varied with EEG montage. The maximum source localization errors (LE) were 2.5 cm and 1.4 cm for the 64 and 128 electrode configurations respectively. The largest errors occurred for sources nearby the lesion. LE was not systematically smaller with 128 rather than 64 electrodes. Maximum intensity errors were similar for 128 and 64 electrodes. The EEG inverse dipole solution was maximally sensitive to the electrode configuration on the scalp when the source was located deep in the brain (e.g. in the brain stem). We concluded that deep source localization needed an extended sampling of the scalp. PMID- 12085645 TI - Three layer impact simulation for large cranioplasty using non-linear finite element model. AB - In this study, we conducted biomechanical study about implant material and behavior of large cranioplasty upon post-operative impact injury. Previous studies utilized a surface model of the skull and large cranium patch (defect area is 46.7 cm2)[1,2,3,4]. It had single layer with thickness of adult skull as 7 cm in the parietal bone. After construction of the finite element model, the mesh and node data were analyzed for the static behavior of the patch. Four-node and three-node quadrilateral shell elements were mainly used in this model to adapt easily to complex shapes. The overall model consists of 4,740 nodes, 2,168 solid elements, and 4,736 shell elements. Even though, the elements could demonstrate the displacement, stress, and strain by longitudinal along the force direction, subsequent result could not reveal the dynamic behavior of the model. Normal adult skull is consisting of three layers (inner, outer and diploe layer). This structure resembles composite materials that behave differently upon impact. In order to overcome limitation of previous model, we designed physiological model and analyzed dynamic behavior with new implant materials. The composite implant with two popular materials and middle polymer layer was evaluated and compared with bony material. PMID- 12085646 TI - Lesion type misidentification: EEG potential sampling and source reconstruction errors. AB - Accurate EEG source reconstruction needs an appropriate volume conductor head model including, in the presence of a morphological brain lesion, a lesion compartment. Lesion electrical properties (conductivity) can not be measured in vivo and need to be retrieved from literature on the base of lesion type identification, performed by means of diagnostic imaging. However, different pathologies can appear similar at bioimages inspection, leading to uncertain diagnosis and in turn to wrong lesion conductivity assumption. Besides many factors, source reconstruction accuracy depends on lesion conductivity value and on electrodes placing (scalp potential sampling). We investigated the relation between electrodes sampling of the EEG and source reconstruction accuracy in case of uncertain lesion type identification. We adopted an eccentric-spheres head model (including the lesion) and a dipole source. We simulated several pathological conditions considering different electrode montages on the scalp: a "clinical like" sampling with 64 electrodes and an extended sampling with 128 electrodes. Source reconstruction was performed assuming wrong lesion type identification (i.e. introducing an error in the model parametrical setting). We found large errors for both source localization (LE) and source intensity estimations. Maximum LE could be reduced (from 30 to 27 mm) by the extended sampling. Conversely, the mean values of LE mostly increased using more electrodes. The benefit of enlarging the sampling was clear for the intensity estimation. In conclusion, only the most inaccurate source reconstruction could be improved changing electrodes placing; the inherent source reconstruction error due to model parameter setting could not be avoided. PMID- 12085647 TI - Nanocontroller update: building a better artificial neuron. AB - Recent progress in microprocessor design has produced sophisticated 8-bit single chip microcontrollers in small packages. These user-programmable "nanocontrollers", some with as few as eight pins, now include a variety of linear on-chip components. Miniscule complex mixed digital and analog systems are now possible. This paper outlines some of these advances, then describes how using these new microcontroller features to create a better artificial neuron have improved the author's ten-year-old neural network design. PMID- 12085648 TI - An integrated pattern recognition approach for intrusion detection. AB - Intrusion detection systems (IDS) attempt to address the vulnerability of computer-based systems for abuse by insiders and to penetration by outsiders. An IDS is required to examine an enormous amount of data generated by computer networks to assist in the abuse detection process. Thus, there is a need to develop automated tools that address these requirements to assist system operators in the detection of violations of existing security policies. In this research, an automated IDS is proposed for insider threats in a distributed system. The proposed IDS functions as an anomaly detector for insider system operations based on the analysis of the system's log files. The approach integrates dynamic programming and adaptive resonance theory (ART1) clustering. The integrated approach aligns sequences of log events with prototypical sequences of events for performing tasks and classifies the aligned sequences for intrusion detection. The system examined for this research is a Boots System for controlling the movement of boots from one place to another under specific security restrictions related to the boot orders. We present the proposed model, the results achieved and the analysis of an implemented prototype. PMID- 12085649 TI - Biomechanical analysis of padding in child seats and head injury. AB - Head injury is a common finding for infants and young children involved in automobile accidents. Although the child restraint seats have increased the level of safety for the pediatric population, skull fracture and/or brain injury occur during the interaction between the child's head and interior of the car seats with no padding. The introduction of effective and sufficient padding may significantly reduce the head injury. The present study was designed to evaluate the biomechanical effects of padding in child seats to reduce the potential for head injury. A head drop test of a six-month old anthropomorphic dummy was conducted. The side of the dummy head impacted the interior wing of child car seats of relatively soft and stiff materials, and a rigid metal plate at velocities of 2.2, 4.5 and 6.7 m/s. In all tests, three types of padding environments were used (no padding, comfort foam, 16 to 19 mm polypropylene padding). All data were collected at 10 kHz and filtered. A total of 39 tests were conducted. The head injury criteria (HIC), and head acceleration, and head angular acceleration were obtained. The HIC was calculated over a 36 ms interval from the resultant tri-axial acceleration. The angular accelerations were derived from the angular velocity data. The head injury biomechanical parameters decreased with the addition of padding. The HIC, peak acceleration, and angular acceleration were reduced up to 91%, 80%, and 61% respectively. The present results emphasize the importance of energy absorbing padding to provide an improved safety environment in child car seats. PMID- 12085650 TI - Biomechanical analysis of head-neck force in hybrid III dummy during inverted vertical drops. AB - The hybrid III dummy has been used extensively for crash testing. The comparison between the cadaver and dummy data provides the biofidelic nature of dummy head neck system to predict cervical spine injury as a function of applied force. The existing dummy data are limited to a lower drop height up to 0.5 m. The present study quantified the head-neck biomechanical response of the dummy up to a drop height of 1.20 m. At 0.15 m, the head force was 5740 N and 5695 N at the upper neck and 4231 N at the lower neck. At 0.6 m, the head force was 13,000 N and 12,000 N at the upper neck and 8900 N at the lower neck. At 1.2 m, the calculated head force was 19,500 N and 18,600 N at the upper neck and 13,500 N at the lower neck. The present results closely match with data of previous studies. The data indicated that the hybrid III system transmits about 70 to 75% of the applied force from the head or upper neck to the lower neck area. In contrast, the cadaver studies showed for drops from 0.9 to 1.5 m, about 20 to 30% of the applied force was transmitted from the head to the lower neck. The comparison demonstrates the capability of the Hybrid III dummy head-neck force response to realistically predict injury and also the need for comparing the dummy's response to the human head neck force data for injury. PMID- 12085651 TI - Restraints and occupant kinematics in vehicular rollovers. AB - Occupant kinematics and the potential for injury in vehicular rollover crashes are dramatically affected by various restraint system characteristics. This study reviews previous research that utilized various methodologies and test fixtures to evaluate restrained occupant motions, primarily in the vertical direction, during both inverted and quasi-static simulated rollover environments. Additional laboratory tests were conducted in order to understand the static and dynamic excursion of restrained humans and surrogates in typical production motor vehicles under inverted circumstances. These tests indicated that volunteer occupants restrained within a complete vehicle by typical production 3 point seat belts will move vertically towards the vehicle roof structure by approximately 50 150 mm in production vehicles, depending on occupant size and belt configuration, when inverted and subjected to a static 1 g acceleration. Dynamic inverted vehicle drop testing in 3-point belt production vehicles, resulting in 4 to 11 g impacts, resulted in surrogates moving only about an additional 23 to 55 mm beyond the static inverted 1 g excursions. PMID- 12085652 TI - Effects of major histocompatibility complex class II knockout on mouse bone mechanical properties during development. AB - We investigated the effect of major histocompatibility complex class II (MHC II) knockout on the development of the mouse peripheral skeleton. These C2D mice had less skeletal development at 8, 12 and 16 weeks of age compared to wild-type C57BL/6J (B6) male mice. The C2D mice had decreased femur mechanical, geometric and compositional measurements compared to wild type mice at each of these ages. C2D femur stiffness (S), peak force in 3-pt bending (Pm), and mineral mass (Min M) were 74%, 64% and 66%, respectively, of corresponding B6 values at 8 weeks of age. Similar differences were measured at 12 weeks (for which C2D femoral S, Pm and Min-M were 71%, 72% and 73%, respectively, of corresponding B6 values) and at 16 weeks (for which C2D femoral S, Pm and Min-M were 80%, 66% and 61%, respectively, of corresponding B6 values). MHC II knockout delays the development of adult bone properties and is accompanied by lower body mass compared to wild type controls. PMID- 12085654 TI - Modeling a parallel L4 neuron array of the fly (Musca domestica) vision system with a sequential processor. AB - At RMBS 2001 Olson presented a novel approach to image edge detection based on the vision system of the common house fly, Musca domestica [1]. Biologically based vision systems are inherently parallel and the vision related cells form a self-contained cartridge, ommatidium, which is duplicated across the surface of the fly's eye. Histological evidence provides the interconnection both within the vision cartridge and the connections to adjacent cartridges. Due to the parallel nature of biologically inspired vision systems, they outperform computer based digital vision systems in speed performance and memory requirements. Olson provided a model of the cartridge with its intra- and inter-connections. This model, rendered in MATLAB and Excel, demonstrated the feasibility of edge detection in the first several synaptic cellular connections within the cartridge. His results demonstrated how edge detection and object movements are easily obtained using a biologically based vision model. He demonstrated the model using simple rectangular and circular objects. We term this work Olson's Algorithm. We have extended Olson's Algorithm into a high-resolution model using a standard off-the-shelf frame grabber. Although, the frame grabber is a digitally based instrument, its image planes are used to model the photoreceptor layer (R1-R6), the L1, L2 monopolar cell layer, and also the monopolar L4 cell layer. The connections between these cells are programmed in "C". The high resolution model demonstrates the feasibility of using a biologically based vision system in a real world application. Furthermore, it allows object segmentation, movement, and tracking to be modeled prior to implementation in parallel analog hardware. PMID- 12085653 TI - Applying wavefront sensors and corneal topography to keratoconus. AB - We report preliminary results from a study to create custom optics (contact lenses) for persons with the eye disease Keratoconus. Custom optics are intended to correct for higher order aberrations which cannot currently be corrected with conventional contact lenses. A lens design algorithm has been developed that utilizes both wavefront sensor data and corneal topography data. The lenses will be formed from thick, pre-fabricated soft contact lenses using laser ablation. Preliminary ablation of these lenses is demonstrated using a femptosecond Ti: Sapphire (Ti:Al2O3) laser system. PMID- 12085655 TI - A variable valve area, lumped parameter model of left ventricular filling. AB - A variable valve lumped parameter model was developed and validated in porcine trials. The model was used to simulate ventricular and atrial pressures and flows during diastolic filling. The variable valve area model simulations provide reasonable results and take into account other considerations like venous inflow and initial valve aperture. Comparisons between model outputs and in-vivo porcine experiments suggest that the model may be used in the future to assess compliance and effective orifice area based on parameters derived from non-invasive Doppler waveforms. PMID- 12085656 TI - Apatite formation on bioactive zirconium metal prepared by chemical treatment. AB - Bioactive metallic materials, which can directly bond to living bone, are badly needed in dental and orthopedic implants for better long-term results. Forming a layer of bone-like apatite on the surface of the metal is one of the most promising methods to increase its bioactivity. This study creates a new chemical treatment for zirconium metal to induce apatite formation in a simulated body fluid (SBF), and analyzes the apatite layer by using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), and Fourier transform infrared spectroscopy (FTIR). Zirconium samples were soaked in 10% CuCl2 aqueous solution at 80 degrees C for 72 hours, heat-treated at 500 degrees C for one hour and soaked in SBF for various periods. An apatite-like layer appeared on the zirconium surface as early as three days in SBF, and increased with immersion time. EDS and FTIR confirmed that the deposited layer was apatite. This implies that bioinert zirconium metal can become bioactive by forming an apatite layer in a simulated body fluid after a suitable chemical treatment. The ZrOOH+ hydrogel layer produced on the zirconium surface by CuCl2 and heat-treatments is thought to induce the apatite formation. PMID- 12085657 TI - L-glutamic acid production in a continuous stirred tank bioreactor using coimmobilized bio-catalyst using a fluorosensor. AB - The production of L-Glutamic acid has been studied using coimmobilized whole cells of pseudomonas reptilivora and micrococcus glutamicus in a two litre Tokyo Rikakikai fermentor using glucose as selected production medium. The process was carried out at an optimum temperature of 32 degree Celsius and a pH of 7.2. The progress of the reaction was recorded using Dr. Ingold fluorosensor. The effect of initial substrate concentration, speed of agitation, volume ofcalcium alginate beads and aeration rate on the yield of glutamic acid has been investigated. It has been found that the acid production increases exponentially with substrate concentration, and mass transfer co-efficient varied linearly with aeration rate. The kinetic parameters also had been estimated. PMID- 12085658 TI - Biodegradable synthetic polymer scaffolds for reinforcement of albumin protein solders used for laser-assisted tissue repair. AB - Laser tissue soldering has been investigated for several years by researchers in our laboratory as an alternative to conventional tissue fasteners, including sutures, staples and clips. Laser tissue soldering is a bonding technique in which protein solder is applied to the tissue surfaces to be joined, and laser energy is used to bond the solder to the tissue surfaces. Over the past four years we have been investigating the use of synthetic polymer membranes as a means for reinforcing the strength of tissue repairs formed using traditional laser tissue soldering techniques. The purpose of this study was to assess the influence of various processing parameters on the strength of tissue repairs formed using the reinforced solder. Biodegradable polymer membranes of specific porosity were fabricated by means of a solvent-casting and particulate-leaching technique, using three different poly(alpha ester)s: polyglycolic acid (PGA), polylactic acid (PLA) and poly(L-lactic-co-glycolic acid) (PLGA). In addition, several membranes were also prepared with poly(ethylene glycol) (PEG). The membranes were then doped with the traditional protein solder mixture of serum albumin and indocyanine green dye. Varied processing parameters included the polymer type, the PLGA copolymer blend ratio, the polymer/PEG blend ratio, the porosity of the polymer membrane and the initial albumin weight fraction. Variation of the polymer type had negligible effect on the strength of the repairs. Although it is known that alteration of the copolymer blend ratio of PLGA influences the degradation rate of the polymer, this variation also had no significant effect on the strength of the repairs formed. Increased membrane flexibility was observed when PEG was added during the casting stage. An increase in the porosity of the polymer membranes led to a subsequent increase in the final concentration of protein contained within the membranes, hence aiding in strengthening the resultant repairs. Likewise, an increase in the initial albumin weight fraction increased the strength of the resultant repairs. PMID- 12085659 TI - A novel paradigm for telemedicine using the personal bio-monitor. AB - The foray of solid-state technology in the medical field has yielded an arsenal of sophisticated healthcare tools. Personal, portable computing power coupled with the information superhighway open up the possibility of sophisticated healthcare management that will impact the medical field just as much. The full synergistic potential of three interwoven technologies: (1) compact electronics, (2) World Wide Web, and (3) Artificial Intelligence is yet to be realized. The system presented in this paper integrates these technologies synergistically, providing a new paradigm for healthcare. Our idea is to deploy internet-enabled, intelligent, handheld personal computers for medical diagnosis. The salient features of the 'Personal Bio-Monitor' we envisage are: (1) Utilization of the peripheral signals of the body which may be acquired non-invasively and with ease, for diagnosis of medical conditions; (2) An Artificial Neural Network (ANN) based approach for diagnosis; (3) Configuration of the diagnostic device as a handheld for personal use; (4) Internet connectivity, following the emerging bluetooth protocol, for prompt conveyance of information to a patient's health care provider via the World Wide Web. The proposal is substantiated with an intelligent handheld device developed by the investigators for pediatric cardiac auscultation. This device performed accurate diagnoses of cardiac abnormalities in pediatrics using an artificial neural network to process heart sounds acquired by a low-frequency microphone and transmitted its diagnosis to a desktop PC via infrared. The idea of the personal biomonitor presented here has the potential to streamline healthcare by optimizing two valuable resources: physicians' time and sophisticated equipment time. We show that the elements of such a system are in place, with our prototype. Our novel contribution is the synergistic integration of compact electronics' technology, artificial neural network methodology and the wireless web resulting in a revolutionary new paradigm for healthcare management. PMID- 12085660 TI - Challenges in designing a multi-function patient monitor. AB - This article presents a discussion of some challenges in the design of a multi function patient monitor. The term "multi-function patient monitor" refers to a single medical device that monitors multiple physiological parameters from the same patient. A common configuration of the monitor is a personal computer (PC) platform communicating with several modules each designed to monitor one physiological parameter. Although such an integration of modules allows the modules to access to the display screen, keyboard, power supply and printer of the PC, the integration also involves new design challenges. The foremost challenge is the increased demand on the PC's resources, while another challenge is compatibility among the various modules. Examples of compatibility are electromagnetic compatibility, and clinical compatibility. A well-known example of incompatibility is the non-invasive blood pressure monitor cuff cutting off blood flow to the finger with a pulse oximeter probe, resulting in a Low Signal alarm in the pulse oximeter. Other challenges include clutter on the display screen and confusing alarm annunciation. This article discusses these design challenges and proposes some solutions based on engineering principles, and common sense considerations. Examples will also be drawn from case reports available in the public domain. PMID- 12085661 TI - A method for assessing balance control in rodents. AB - Recent research has shown that after spinal cord injury, the nervous system reorganizes. Nevertheless, little is known of the effects of neural reorganization, or plasticity, on motor skills. In this work, we present a method that utilizes kinetic and kinematic analysis, for investigating balance control in a rodent model of incomplete spinal cord injury. In this setup, the animals sit unconstrained on their hindlimbs on a platform while they eat a Fruitloop. In this posture, the animal is supporting all the body weight on its hindquarters removing the need for the animal to support itself on its hindlimb or maintain appropriate forelimb-hindlimb coordination for functional gait. The platform is bolted to a force transducer to provide measurements of shear forces in orthogonal directions in the horizontal plane. Reflective markers on the hip and tail of the animal indicate sway of the animals body. Using this method the effect of extensive 12 week long treadmill locomotor training on balance control in rodents with incomplete thoracic spinal cord contusion injury (iSCI) was assessed. One iSCI rat did not undergo training, while a sham injured and 3 other iSCI rats underwent training. The shear forces and sway produced by the injured untrained rat were much larger than for the sham or the injured but trained rats. Stance width assessed from overground walking in a separate study was also larger in the injured untrained rat. These data suggest that balance control may be improved by a locomotor training paradigm. This simple method for assessing balance control could thus be utilized in longitudinal studies to evaluate the effectiveness of pharmacological and locomotor therapies for repair and recovery after spinal cord injury. PMID- 12085662 TI - The effects of vitamin E on the proliferation and viability of irradiated fibroblast-like cells. AB - Few studies have indicated the use of antioxidants in the protection against cancers. The goal of this study was to develop a tissue culture model using MRC-5 cells, which would allow for rapid testing of treatments for skin damage, mediated by exposure to ultraviolet radiation (UVR). Overall the specific aims were to evaluate the morphological and pathophysiological responses of MRC-5 fibroblast cells exposed to UVR and antioxidant supplementation. In the first phase, MRC-5 cells were exposed to UVR for 30, 45 or 60 minute time intervals, and cellular damage was assessed biochemically and morphologically. In the second phase, cells were treated for 45 minutes in the presence or absences of physiological levels of vitamin E. The cells were evaluated after 24, 48, and 72 hours for biochemical and morphological changes. Structural changes were noted throughout the experimental phase among the UVR exposed groups compared with native control. However, treatment with vitamin E pre and post exposure provided a protective role by demonstrating lower levels of MDA (p < 0.05) as well as normal structural appearance. PMID- 12085663 TI - Release of inflammatory cytokines by macrophages and synovial cells challenged with tumor necrosis factor. AB - Late aseptic loosening of total joint implants continues to be a common cause of implant failure. However, the pathophysiology of implant loosening remains controversial as to which factors at the implant tissue interface plays a crucial role in implant failure. The most prominent features of the foreign body membrane obtained from patients undergoing revision hip surgery were the presence of lymphocytes, histiocytes, giant cells, and immature collagen formation. The inflammatory sites are often characterized by the infiltration of activated lymphocytes and macrophages into the synovial membrane followed by proliferation of the synovial cells. The present study was conducted to determine if synovial cells in addition to macrophage cells are activated by tumor necrosis factor (TNF), which ultimately leads to foreign body membrane proliferation and ultimately joint space destruction. Macrophages or synovial cells were seeded at a density of 1 x 10(6) cells/ml and 5 ml were placed onto a 12 mm culture dish. The cells were challenged with either tissue culture media or media containing 2 ng/ml LPS or various doses of TNF (0.5, 5 and 50 ng/ml). The result indicated that both cell types were able to produce the inflammatory cytokines (IL-1 and IL 8 as early as 8 hours after a challenge with peak production occurring between 18 24 hours). The data suggest that both cell types are capable of eliciting inflammatory mediators, which can ultimately lead to joint destruction. PMID- 12085664 TI - Amelioration of experimental allergic encephalitis (EAE) through phase 2 enzyme induction. AB - The multiple sclerosis (MS) lesion is characterized by an inflammatory cell mediated attack on white matter. Oxidative stress appears to play a role in the onset and progression of MS. We reasoned that decreasing oxidative stress might ameliorate MS. One way of decreasing oxidative stress is to induce phase 2 enzymes. The model chosen to test this hypothesis was experimental allergenic encephalomyelitis (EAE) induced in the Lewis rat. The 26 animals were placed into two groups: 1) those on normal rat chow, 2) those on rat chow containing 250 mumoles t-butylhydroxyanisole (BHA)/kg. After 2 weeks, animals were administered 100 micrograms guinea pig myelin basic protein and examined daily in a blinded fashion. Twenty-nine days later, animals were sacrificed, blood collected for glutathione (GSH) measurements and tissues collected for histology. Six of the 13 control chow animals developed hindlimb weakness or paralysis while 5 developed tail weakness only. Only 1 BHA fed animal exhibited symptoms--hindlimb weakness. Clinical symptoms correlated well with the extent of perivascular lymphocyte infiltration. Animals with BHA in the diet had 20% higher red cell GSH indicting induction of phase 2 enzymes. We conclude that dietary phase 2 enzyme inducers should be examined for their ability to ameliorate MS. PMID- 12085665 TI - Localization of cytokines in heart ventricular and apex tissues exposed to sustained delivery of AED, T, and DHT using a rat model. AB - Studies have shown that endogenous estrogen minimized cellular injury at the organ level; however, very little research was done to determine the effects of endogenous androgens such as Testosterone (T), Dihydrotestosterone, (DHT), and Androstenedione (AED) on the cardiovascular system at the cellular level Studies targeted at establishing such effects will broaden our understanding of the roles played by these male steroid hormones on the cardiovascular system. Our objective therefore was to (1) Use a Rat model and sustained delivery of physiological levels of these hormones to (1) evaluate pathophysiological effects on the cardiovascular system; and (2) localized cytokines such as IL-1, IL-6 and TNF on these tissues. Four groups of Sprague Dawley rats were included in the study. Group 1 animals served as control, groups II, III, and IV were treated with TCPL drug delivery devices containing 40 mg each of T, DHT, and AED, respectfully. Animals were sacrificed after 90 days of exposure. The ventricles and apex were immunostained for IL-1, IL-6, TNF and were also stained with Hemotoxylin and Eosin for histopathological evaluation. Results showed that TCPL drug delivery systems released 5 ng/ml/day of T, and 2 ng/ml/day of DHT and AED. IL-6 was expressed in the control heart ventricle and T, DHT, and AED reduced expression of the cytokine significantly after 90 days. Very small number of cells from the apex responded to IL-6 than cells within the ventricles. The results revealed that (i) the exposure of sustained levels of androgenic hormones exhibited myocardial hypertrophic condition compared to the control animals, (ii) the control animals had a two-fold increase in the ventricles IL-6 production over T treated animals and approximately five-fold increase over DHT and AED treated animals, and (iii) other inflammatory cytokines IL-1 and TNF were not differentially expressed in the ventricles of experimental animals when compared with the control; and (iv) In the apex tissue, animals treated with AED exhibited cells with increased response to IL-6 and a decrease cells responding to TNF and IL-1. IL-6 production in the ventricles decreased in animals exposed to sustained androgenic steroids and this decrease could possibly lead to increased blood flow, resulting in better oxygenation of the myocardium and improved cardiac function. PMID- 12085666 TI - [Plasmodium protein-kinases and control of cellular proliferation: molecular targets for new antimalarial drugs]. PMID- 12085667 TI - [Efficacy and toxicity of aminoglycoside therapy in the elderly: combined effect of both once-daily regimen and therapeutic drug monitoring]. AB - This study was aimed to compare with previous results (Grillot et al., 1994), the efficacy of amikacin adaptive optimal control in a geriatric hospital. PATIENTS: During six months, 32 patients (aged of 82 +/- 8 years) were included versus 51 during two years (aged of 80 +/- 5). The mean age was not different between the two populations (NS, Student test). They received amikacin initial dosage of 17.7 +/- 5.1 mg/kg/d (vs 13.3 +/- 3.5 for the reference study) and maintenance dosage of 15.1 +/- 4.8 mg/kg/d (vs 11.8 +/- 5.1 for the reference study). METHOD: Two efficacy outcomes (E1 and E2) and 1 toxicity outcome (T) were taken into account: E1 estimated the effect of adaptive control on maximal drug level, E2: overall recovery. Toxicity outcome was used: T the nephrotoxicity (increasing creatininemia over 44 mumol/l). RESULTS: All the results are given versus the reference study. 57.6% versus 29.4% of adaptive strategy were once-a-day. E1: Chi square test show that initial dosage and maintenance dosage are greater our study than the previous one (p < 0.05: 78.8% versus 5.9% for initial dosage, 84.4% versus 13.8% for maintenance dosage). E2: 73.6% overall of recovery versus 77% (NS, Chi square test). T: 94% versus 85% (p < 0.05, Chi square test) of creatininemia variation are lower than 44 mumol/l. Duration of treatment is 9.8 +/- 4.8 versus 15 +/- 9 days (p < 0.5, Student test). CONCLUSIONS: Once-a-day strategy in amikacin therapeutic regimen is no more efficient but decreases toxicity and duration treatment. PMID- 12085668 TI - [Insulin-dependent neonatal and infant diabetes: genetics and physiopathology]. AB - Insulin-dependent neonatal diabetes (ND) mellitus is uncommon with a frequency of 1/500,000 neonates in Europe. ND is characterised by hyperglycaemia, very low or undetectable insulin levels associated with intrauterine growth retardation and malformations. HLA haplotypes of juvenile diabetes or autoimmunity are not present in ND patients. Sporadic and familial forms are observed. ND could be persistent (PND) or transient (TND). Diabetes relapses occur in approximately 40% of TND patients. Hypothesis for ND aetiology such as pancreatic or beta pancreatic islets of Langerhans immaturity or abnormalities of pancreas organogenesis are postulated. Different genetic basis underlie transient or permanent forms though their clinical features do not allow to distinguish them. TND may in about 20-30% of the cases be associated with chromosome 6 paternal uniparental disomy. A candidate locus for an imprinted gene is mapped to 6q24. The permanent forms are less understood. Homozygous mutations of the IPF1/PDX1 (MODY4) and of the Glucokinase (GK, MODY2) genes have been reported. The association of a ND with a macroglossia should be a strong indicator for genetic testing. The genetic findings of a paternal disomy uniparental allows the prediction of a transient rather than a permanent form. Mutation in the Glucokinase gene should be sought in an infant with ND whose first degree relatives have glucose intolerance. PMID- 12085669 TI - [Immunological diagnosis of scleroderma]. AB - Systemic sclerosis is characterized by a fibrosis and a microvascular injury. These vascular lesions can affect internal organs causing severe visceral damages. The pathogenesis of this disease is complex but some immunological disorders with a production of antinuclear auto-antibodies can be useful to the clinicians, especially for an early diagnosis and to distinguish the scleroderma/myositis overlap syndromes. This review presents the target antigens and the methods of detection of the different antinuclear auto-antibodies. Sensitivity, specificity and clinical association between the antinuclear auto antibodies and the different subclasses of scleroderma are also presented in detail. PMID- 12085670 TI - [Prognostic factors in myelodysplasia syndromes]. AB - Large studies have shown that, in MDS, main prognostic factors for survival and progression to AML were the percentage of bone marrow blasts, the number and importance of cytopenias, and the presence of cytogenetic abnormalities. The combination of those 3 factors has yielded an International Prognostic Scoring System (IPSS), with very strong prognostic value for survival and progression to AML, which can therefore be used as a guideline for therapeutic choices. Other important prognostic factors can be derived from molecular studies. They include ras genes mutations, p53 mutations and p15 hypermethylation, but their independent prognostic value remains uncertain. PMID- 12085671 TI - [Physiopathology of myelodysplastic syndromes]. AB - Myelodysplastic syndromes are clonal diseases of the hematopoietic stem cell with normal or increased bone marrow cellularity and peripheral cytopenias. Pathophysiology of these diseases is complex with frequent ras mutations, a growth defect of immature progenitors mainly erythroid progenitors, and increased apoptosis of differentiated cells. This growth defect could be linked to (1) a resistance to hematopoietic cytokine stimulation although, erythropoietin receptor expression and functionality are normal and/or (2) increased susceptibility to apoptosis due to overexpression of the death domain receptor Fas on CD34+, CD33+ and GPA+ cells. Stromal cells are thought to produce increased quantities of inhibitory cytokines such as TNF-alpha, TGF-beta, IFN gamma et IL-1. Better understanding of MDS pathophysiology is required for applying adequate therapy either blocking apoptosis or stimulating hematopoiesis. PMID- 12085673 TI - [Childhood myelodysplastic syndromes]. AB - Myelodysplastic syndromes (MDS) in children constitute a heterogeneous disorder, including "primary" MDS and MDS associated with constitutional disorders or metabolic diseases. The Franco-American-British (FAB) cytological classification for adults can be applied for childhood in 50 to 100% of the cases. The transformation into acute myeloblastic leukemia often occurs, but stabilisation or spontaneous regression of the disease may also be observed. Allogenic bone marrow transplantation is the best curative option when treatment is necessary. PMID- 12085672 TI - [The place of growth factors in the treatment of myelodysplastic syndromes]. AB - The myelodysplastic syndromes (MDS) are clonal hematological disorders characterized by ineffective hematopoiesis manifested by anemia, neutropenia, thrombocytopenia or a combination. Correction of these cytopenia is a priority in MDS without excess of blasts. Treatment of anemia depends mainly on erythrocyte transfusions. However with the ability of recombinant human hematopoietic growth factor many trials have been promoted. In vitro, erythroid progenitors from MDS patients are able to differentiate but they require much higher concentrations of erythropoietin than normal progenitors. Trials using rHu-Epo alone are disappointing. Combining rHu-Epo and rHu-G-CSF induces more encouraging results showing a synergistic effect particularly clear in sideroblastic anemia. Patients with low endogenous Epo level and low transfusion need are more likely to respond. Clinician should be able in the future to identify MDS patients with a chance of reversal of anemia or transfusion dependency. PMID- 12085674 TI - [New WHO classification of myelodysplastic syndromes. The consequences]. AB - Myelodysplastic syndromes (MDS) are clonal hematopoietic disorders. Diagnosis and classification of MDS are based on morphological features of blood and bone marrow cells. The F.A.B. classification is widely used, but remains unclear in some cases of MDS. A World Health Organization (WHO) working group proposed a new classification of MDS, to improve the homogeneity of the different categories in term of physiopathology and prognosis. The new WHO classification incorporated some clinical and etiological data, and the results of cytogenetical or molecular analysis. The homologies and the differences between these two classification are showed, and the clinical consequences of the new classification are discussed. PMID- 12085675 TI - Women's mental health: focus on sexual and reproductive issues. PMID- 12085676 TI - Female sexual disorders: psychiatric aspects. AB - OBJECTIVE: This manuscript reviews the current information concerning female sexual dysfunction that is relevant to general psychiatric practice. METHOD: Research identified by the key words sexual dysfunction and prevalence, comorbidity, psychiatric drugs, or pharmacotherapy is reviewed. RESULTS: Epidemiologic studies indicate that approximately 30% of female subjects between ages 18 and 59 years have sexual complaints of at least 3 months' duration in the past year. A high comorbidity with other psychiatric syndromes exists. Many psychiatric drugs are associated with sexual dysfunction. Drug treatments for female sexual dysfunction are being investigated. CONCLUSION: Knowledge concerning the treatment of female sexual dysfunction is important to the general psychiatric clinician. PMID- 12085677 TI - Managing bipolar disorder during pregnancy: weighing the risks and benefits. AB - BACKGROUND: Challenges for the clinical management of bipolar disorder (BD) during pregnancy are multiple and complex and include competing risks to mother and offspring. METHOD: We reviewed recent research findings on the course of BD during pregnancy and postpartum, as well as reproductive safety data on the major mood stabilizers. RESULTS: Pregnancy, and especially the postpartum period, are associated with a high risk for recurrence of BD. This risk appears to be limited by mood-stabilizing treatments and markedly increased by the abrupt discontinuation of such treatments. However, drugs used to treat or protect against recurrences of BD vary markedly in teratogenic potential: there are low risks with typical neuroleptics, moderate risks with lithium, higher risks with older anticonvulsants such as valproic acid and carbamazepine, and virtually unknown risks with other newer-generation anticonvulsants and atypical antipsychotics (ATPs). CONCLUSIONS: Clinical management of BD through pregnancy and postpartum calls for balanced assessments of maternal and fetal risks and benefits. PMID- 12085678 TI - The role of estrogen in schizophrenia: implications for schizophrenia practice guidelines for women. AB - OBJECTIVE: The objective of this paper is to integrate what is known about estrogen effects on symptoms and treatment response into a global understanding of schizophrenia. The aim is to expand Canadian schizophrenia guidelines to include the specific needs of women. METHOD: We searched the Medline database; keywords included estrogen, estrogen replacement therapy, schizophrenia, psychosis, treatment, tardive dyskinesia (TD), and women. We examined reference lists from relevant articles to ensure that our review was complete. We review the evidence for the effects of estrogen in schizophrenia and we make recommendations for the next revision of official practice guidelines. RESULTS: The epidemiologic evidence suggests that, relative to men, women show an initial delay in onset age of schizophrenia, with a second onset peak after age 44 years. This points to a protective effect of estrogen, confirming animal research that has documented both neurotrophic and neuromodulatory effects. Clinical research results indicate that symptoms in women frequently vary with the menstrual cycle, worsening during low estrogen phases. Pregnancy is often, though not always, a less symptomatic time for women, but relapses are frequent postpartum. Some work suggests that in the younger age groups women require lower antipsychotic dosages than men but that following menopause they require higher dosages. Estrogen has been used effectively as an adjunctive treatment in women with schizophrenia. Estrogen may also play a preventive role in TD. CONCLUSIONS: Symptom evaluation and diagnosis in women needs to take hormonal status into account. Consideration should be given to cycle-modulated neuroleptic dosing and to careful titration during pregnancy, postpartum, and at menopause. We recommend that discretionary use of newer neuroleptic medication and adjuvant estrogen therapy be considered. PMID- 12085679 TI - Should psychologists be granted prescription privileges? A review of the prescription privilege debate for psychiatrists. AB - BACKGROUND: The debate over whether clinical psychologists should be granted the right to prescribe psychoactive medication has received considerable attention over the last 2 decades in the US, but there has been relatively little discussion of this controversial topic among Canadian mental health professionals, namely psychologists and psychiatrists. Proponents of prescription privileges (PPs), including the American Psychological Association (APA), argue that psychologists do not and cannot function as independent professionals because the medical profession places many restrictions on their practice. It is believed that PPs would help circumvent professional psychology's impending marginalization by increasing psychology's scope of practice. Proponents also argue that PPs would enhance mental health services by increasing public access to professionals who can prescribe. OBJECTIVE: The purpose of this article is to inform psychiatrists about the major arguments presented for and against PPs for psychologists and to discuss the major implications of PPs for both professional psychology and psychiatry. METHODS: We conducted a literature search of relevant articles published from 1980 to the present appearing on Psychlit and Medline databases, using "prescription privileges" and "psychologists" as search titles. CONCLUSION: Although proponents present several compelling arguments in favour of PPs for psychologists, pilot projects relating to feasibility and efficacy are either sparse or incomplete. Thus, it is too soon to tell whether PPs could or should be pursued. Clearly, more research is needed before we conclude that PPs for psychologists are a safe and necessary solution to psychology's alleged impending marginalization. PMID- 12085680 TI - Experiments in change: pretrial diversion of offenders with mental illness. AB - OBJECTIVE: Our objective was to study the outcomes experienced by 2 communities after implementing pretrial diversion of offenders with mental illness. METHOD: The same method of diversion was implemented in a predominately urban and a predominantly rural county. We collected retrospective clinical and offence data from pretrial diversion assessments conducted in court. As well, we measured outcome for the diversion procedure in terms of actual vs expected rates of recidivism. RESULTS: Prior psychiatric treatment was associated with the diverted group, and a criminal history was associated with the nondiverted group. In the larger, urban county the diversion option was offered more often to persons with psychoses, mood disorders, and minor offenses. Conversely, in the smaller rural county diversion was offered most often to persons accused of serious offenses. The recidivism found in urban and rural diverted groups after a year of supervised care was only 2% to 3%, but the rate of use of diversion in both counties was low, owing to selection biases. CONCLUSION: Pretrial diversion of offenders with mental illness accused of minor crimes is eminently feasible for both urban and rural settings, provided that police, crown, and treatment policies are coordinated to favour the treatment option rather than prosecution. PMID- 12085681 TI - Prevalence and correlates of elder abuse and neglect in a geriatric psychiatry service. AB - OBJECTIVE: To determine the prevalence and correlates of 4 types of elder abuse and neglect in a geriatric psychiatry service. METHOD: We conducted a cross sectional retrospective chart review of new in- and outpatients seen by the Montreal General Hospital Division of Geriatric Psychiatry in one calendar year. RESULTS: Abuse or neglect was suspected or confirmed in 20 (16%) of 126 patients, comprising financial abuse in 16 (13%), neglect in 7 (6%), emotional abuse in 5 (4%), physical abuse in 3 (2%), and multiple abuse in 7 (6%). On bivariate analysis, patients living with nonspouse family, friends, or other persons were significantly more likely to have suffered abuse than were those living with their spouse or in a supervised setting (OR 10.5; 95%CI, 2.3 to 47.8); widowed, divorced, or separated patients were significantly more likely to have suffered abuse than were married patients (OR 4.7; 95%CI, 1.02 to 22.0). Nonsignificant trends included female sex (OR 4.1; 95%CI, 0.89 to 18.6); alcohol abuse (OR 2.1; 95%CI, 0.71 to 6.2); behaviour problems (OR 1.9; 95%CI, 0.71 to 5.2); and chronic cognitive impairment (OR 1.4; 95%CI, 0.55 to 3.8). Although living situation with nonspouse family, friends, or others and marital status of widowed, divorced, or separated were significantly associated with abuse when examined in separate logistic regression models, both were nonsignificant when examined together, suggesting collinearity. Both were retained in the model because they probably represent different aspects of vulnerability. The final model included living situation with nonspouse family, friends, or others (OR 6.1; 95%CI, 0.75 to 49.5) and widowed, divorced, or separated marital status (OR 2.4; 95%CI, 0.21 to 26.8). Nonsignificant trends included female sex (OR 2.6; 95%CI, 0.45 to 14.4); alcohol abuse (OR 2.2; 95%CI, 0.59 to 7.9); and lowest quartile on the Global Assessment of Functioning (GAF) scale (GAF < 35; OR 2.0; 95%CI, 0.64 to 6.0). CONCLUSIONS: The practical implications of our study are that elder abuse and neglect are common among patients referred to geriatric psychiatry services, that such services should have access to multidisciplinary expertise and resources to deal with abuse, and that certain situations may signal higher risk. In our setting, the situation of living with nonspouse family, friends, or other persons in a nonsupervised setting and a history of family disruption by widowhood, divorce, or separation were significant correlates of abuse. Suggestive but nonsignificant trends of potential importance (OR > or = 2.0) included female sex, alcohol abuse, and lowest quartile of functional status. Study limitations include a cross-sectional retrospective chart review design, a clinically derived sample, a small sample size, and a lack of structured instruments for several variables. PMID- 12085682 TI - Occupational effects of stalking. AB - OBJECTIVE: This case report and discussion describe the psychiatric and social consequences of being a stalking victim, with particular focus on its impact on the victim's occupation. METHOD: Data were gathered from the assessment and arbitration hearing of a female employee who lost her job while being stalked. Computerized literature searches were used to identify relevant papers from psychiatric and legal journals. RESULTS: This case illustrates many of the common features of stalking. The female victim was harassed by a male after a failed intimate relationship. The victim suffered from depression, anxiety, guilt, shame, helplessness, humiliation, and posttraumatic stress disorder (PTSD). The stalking affected her psychological, interpersonal, and occupational functioning. Consequently, she was fired for poor work performance and poor attendance. CONCLUSION: Stalking may affect a victim's ability to work in several ways. The criminal behaviours often interfere directly with work attendance or productivity and result in the workplace becoming an unsafe location. Further, stalking may indirectly affect a person's ability to work through the many adverse emotional consequences suffered. PMID- 12085683 TI - Gender-role conflict and suicidal behaviour in adolescent girls. AB - OBJECTIVE: This study examined whether gender-role conflict influenced the suicidal behaviour of adolescent girls. METHOD: We designed a checklist and used it to perform a chart review. RESULTS: Gender-role conflict was associated with suicidal behaviour in 26.79% of the adolescent girls, and 2.68% of the adolescent boys, that we studied. CONCLUSIONS: Gender-role conflict plays an important role in the suicidal behaviour of girls. At present, there is no published research examining the impact of gender-role conflict on suicidal adolescents. PMID- 12085685 TI - Assessing and managing compulsive scratching in schizophrenia with chronic renal failure. PMID- 12085684 TI - Gabapentin treatment of impulsive-aggressive behaviour. PMID- 12085686 TI - Using the rating scale for psychotic symptoms to characterize delusions expressed in a schizophrenia patient with "Internet psychosis". PMID- 12085687 TI - The ward changes address: an entire hospital department moves to a modern building. PMID- 12085688 TI - Sildenafil citrate for female orgasmic disorder. PMID- 12085689 TI - Suicide among immigrants to Canada from the Indian subcontinent. PMID- 12085690 TI - Fire fetishism in a female arsonist? PMID- 12085691 TI - Early access defibrillation: a public health issue for all ages. PMID- 12085692 TI - Views from the AMA. J. Edward Hill, MD. PMID- 12085693 TI - Dengue fever in mid-Missouri. A case report. PMID- 12085694 TI - Medication and the elderly: avoiding the pitfalls of polypharmacy. PMID- 12085696 TI - Inside John Forbes Nash Jr.'s beautiful mind. PMID- 12085695 TI - Natriuretic peptides. AB - Thus the natriuretic peptides represent an important class of molecules in patients with congestive symptoms. As such, natriuretic peptide measurements can assist in the evaluation of patients with heart failure or to exclude this as a cause in patients with dyspnea. Increasing the levels of natriuretic peptides may offer an important therapeutic advance in patients with heart failure. PMID- 12085697 TI - Future surgeons. PMID- 12085698 TI - [Informative value of adenosine deaminase and 2-deoxyadenosine deaminase in the diagnosis of tuberculous pleurisy]. AB - Adenosine deaminase (ADA) and 2-deoxyadenosine deaminase (2-deoxyADA) activities were measured in pleural exudation of different origin in 373 patients (124 of these with serous tuberculous pleurisy and 75 with specific empyema). High diagnostic value of ADA activity in the presence of threshold value higher than 35 Units/liter was established: the sensitivity of the test in the diagnosis of tuberculous pleurisy and empyema was 94.4 and 93.3% respectively, specificity 93.0 and 89.5%, respectively. Judging by the coefficient of 2-deoxyADA/ADA, total ADA activity is determined by the activity of ADA-2 isoenzyme, while in tuberculous empyema by ADA-1. Additional use of this coefficient, requiring measurement of 2-deoxyADA activity, does not improve the efficiency of the diagnostic process, including measurement of total ADA activity in pleural exudate. PMID- 12085699 TI - [Chromatographic analysis of low molecular weight fraction of cerebrospinal fluid in children with acute neuroinfections]. AB - Low molecular-weight (oligopeptide) fraction of the cerebrospinal fluid was analyzed by high-performance reversed phase liquid chromatography in 30 children with bacterial and viral neuroinfections. The incidence and height of chromathoraphic peaks in bacterial meningitis depended on the disease etiology, stage, and severity. Qualitative and quantitative composition of low molecular weight fraction of the liquor varied in patients with viral neuroinfections, depending on the severity of the cerebral parenchyma involvement. Differences in chromatographic profiles in complicated and uneventful course of neuroinfections indicate a possible damaging, protective, or regulatory effect of the liquor peptides. These data focus the attention on the role of oligopeptides in the genesis of neuroinfectious process, significance of search for peptide markers, their further isolation, identification, and development of test systems available for clinical application. PMID- 12085700 TI - [High performance liquid chromatography and radioimmunoassay in analysis of serum hydrocortisone concentration and pharmacokinetic parameters of glucocorticoids in human. The choice of method for clinical practice]. AB - Serum hydrocortisone concentrations were measured by high performance liquid chromatography (HPLC) and radioimmunoassay (RIA) in 24 children with different diseases and 16 healthy adults. HPLC and RIA proved to be equally sensitive. However the authors recommend HPLC as a reliable, rapid, and ecologically pure method, which is, in addition, selective and economic. PMID- 12085701 TI - [Total antioxidant activity of the lacrimal fluid]. AB - Total antioxidant activity of the lacrimal fluid (TAALF) was studied by luminol dependent iron-induced chemiluminescence in an original model system on citrate phosphate buffer. Total antioxidant activity was estimated in percent of control (light sum of the model system fluorescence). The mean decrease of the light sum of the model system fluorescence in the presence of normal human lacrimal fluid supernatant was more than 50%, in glaucoma 67.2 +/- 4.43%, in cataract 58.5 +/- 2.61%, and in uveitis 76.9 +/- 1.13% of control. Traditional therapy of uveitis was associated with an increase of TAALF. Use of the proposed method for TAALF evaluation together with clinical data allows justified and individual antioxidant treatment of patients with ocular diseases and helps monitor the treatment efficiency. The study is noninvasive, cheap, and simple, which recommends evaluation of TAALF in clinical practice as a rapid test indicating the level of free radical oxidation in ocular tissues. PMID- 12085702 TI - [Laboratory techniques for diagnosis and monitoring of diabetes mellitus (Lecture)]. PMID- 12085703 TI - [Double bonds of fatty acids, alcohols glycerol, cholesterol and nonpolar serum lipids. Diagnostic value of hypercholesterolemia]. AB - A method for evaluation of double bonds (DB) by ozone titration in a pool of serum lipids during extraction after Folch is developed. A method for measuring the weight of nonpolar lipids in apoB-100-lipoproteins (LP) with a hydrophobic fluorescent probe K-37 was used. Cholesterol and glycerol alcohols were measured in the serum using Cholesterol and Triglycerides kits (Hoffmann la Roche). Twenty four patients with myocardial infarction, 29 coronary patients, and 37 healthy children were examined. The level of cholesterol alcohol reflects the number of double bonds in fatty acids (FA) and hence, the content of essential polythenic FA in the serum. The higher the level of cholesterol alcohols the greater the pool of essential poly-FA, circulating in the blood as nonpolar cholesterol polyesters within low density lipoproteins (LDL). Presumably, the higher the level of cholesterol LDL, the more pronounced is the deficit of essential poly-FA and the greater is the compensatory activation of the synthesis of endogenous poly-FA. A positive correlation between the content of cholesterol alcohol in the serum and weight of nonpolar lipids in apoB-100-LP was noted. PMID- 12085705 TI - [Methods for detection and characterization of mechanical resistance of erythrocytes in healthy men (review of literature)]. AB - Published reports on methods for evaluation of erythrocyte rheology and mechanical resistance are reviewed. The processes running in red blood cells under the effect of mechanical factors are described. An original method for studies of vibration resistance of erythrocytes is proposed. It allows evaluation of the erythrocyte hemolysis, aggregation, and fragmentation. The method consists in combined vibroexposure of whole capillary blood in vitro (including blood collected under conditions of redistribution of erythropoiesis after exercise) and differentiated cytometry by conductometric hematological scintillators with plotting a histogram of erythrocyte distribution by volume. The method was used by many scientists who obtained different results in examinations of normal subjects. This disagreement of results of different studies is explained by numerous causes, some of which can hardly be taken into consideration. The authors attempted to standardize the method and normal (reference) values. The method is sensitive to various factors (even low intensive) affecting the erythrocyte population and allows the detection of diseases at the preclinical stage and evaluation of the degree of subject's training. PMID- 12085704 TI - [A new approach to the cell pathophysiological study: cell distribution by size and shape as a diagnostic and monitoring tools in illness]. PMID- 12085706 TI - [Reversible aggregation of erythrocytes in children with appendicular peritonitis]. AB - A microtest for quantitative evaluation of the kinetics of reversible aggregation of erythrocytes was used in 47 patients and 203 children with local (121 cases) and diffuse (82 cases) appendicular peritonitis. The patients' ages varied from 3 to 15 years. The diagnosis was confirmed clinically, intraoperatively, and morphologically during examination of removed appendices. The development of appendicular peritonitis of different dissemination was paralleled by pronounced intensification of reversible aggregation of peripheral blood erythrocytes, which suggests high diagnostic and prognostic significance of reversible aggregation of erythrocytes, determines the strategy of preoperative treatment, and helps plan the volume of intervention. PMID- 12085707 TI - [Toxicological screening of biological fluids]. AB - Analysis of the results of evaluation of the complement activity, toxicity of the urine, low molecular weight fraction of the blood, and blood cell hemolysis confirmed the suggested simplified model of endotoxicosis and showed that evaluation of the complement activity and urine toxicity is sufficient for tentative laboratory diagnosis of endotoxicosis. Proceeding from these data it is possible to evaluate the patient's status and predict the disease outcome. Laboratory diagnosis is made using a biological test system and toxicity analyzer. PMID- 12085708 TI - [A method for analysis of biochemical phase of anti-HIV antibody and HIV antigen reaction by release of Ag+-sensitive-SH-containing nonprotein compounds]. AB - The phenomenon of release of Ag(+)-sensitive SH containing protein compounds during reaction of serum antibodies to HIV with HIV antigen glycoproteins (AgHIV) (both reagents taken from diagnostic kits) is described. These compounds were detected in nonprotein fractions of serum anti-HIV + AgHIV reaction media and disappeared after dialysis of nonprotein fractions. On the other hand, this phenomenon was not detected during addition of AgHIV to the serum containing no AgHIV. Presumably, the phenomenon should be referred to secondary concomitant reaction, which seems to be mediated by conformational and/or other processes only during highly specific antigen-antibody intermolecular interactions. That is why this phenomenon seems to characterize the biochemical phase of immune reactions, which can be associated with the denaturing processes as well, running in serum anti-HIV + AgHIV reaction media, and it can be used as an additional test in evaluating anti-HIV antibodies. PMID- 12085710 TI - [Method of quantitative analysis of beta2-microglobulin in human blood]. AB - A simple and economic method of radial immunodiffusion is developed for diagnostic measurements of beta 2-microglobulin in human serum. Human serum and plasma concentrations of beta 2-microglobulin can be measured sufficiently accurately; the results are reproducible. The reproducibility and addictiveness of the method were evaluated. The error of the method is no more than 15% (3-36 mg/liter), which is permissible for methods of this type. PMID- 12085711 TI - [Long-term treatment of depression: there is more than drugs]. AB - Recent studies have emphasized the chronic nature of depressive disorder, and the need for endorsing the same treatment protocols used for other chronic disease, such as diabetes. But duration of treatment does not seem to affect long-term prognosis of patients with depression, once the drug is stopped. Despite treating depression effectively in the short-term, antidepressant drugs may worsen its course. Treatment of depression by pharmacological means is likely to leave residual symptoms in most patients. Such symptoms are important risk factors for relapse. In randomised controlled studies, cognitive behavioural treatment of residual symptoms significantly improved long-term outcome of recurrent depression. Indeed, depressive disorder has a chronic nature, but antidepressants are not the only possible treatment strategy. Cognitive-behavioral psychotherapy is an alternative, which is effective in engendering a complete and lasting recovery in patients with depression. PMID- 12085713 TI - [Therapeutic decision in medical oncology]. AB - The clinical decision in medical oncology is difficult to take. To develop a decisional system based on evidence based medicine, guidelines and protocols could be useful in many complex situations. PMID- 12085712 TI - [Role of chemokine receptors in allergic inflammation and new potential of treatment of bronchial asthma]. AB - The recruitment of T lymphocytes into pulmonary tissue is a critical component of the host response to infections. Migration of T cells into the lung also orchestrates inflammation, tissue injury and remodelling of tissue architecture associated with the principal chronic inflammatory pulmonary diseases such as asthma. Chemotactic cytokines, named chemokines, play a major role in regulating localization of Th2 cells and other leukocytes into the lung during an asthmatic attack. Various proinflammatory functions have been ascribed to chemokines going from chemotaxis, activation and degranulation of distinct leukocyte subsets to remodelling of inflammed pulmonary tissue. This paper focuses on recent data either from clinical observations or animal models that have highlighted the importance of chemokines in pulmonary allergic inflammation. The recent advances in this field may now lead to the development of novel therapies for the allergic pulmonary diseases. PMID- 12085714 TI - [Clinico-microbiological comments on various cases of candidemia]. AB - During a 3 year period 48 patients with one or more blood cultures positive for Candida spp. were enrolled in the study. One patient presented 3 consecutives episodes of candidemia with infection of the port a cath. Fifty cases of candidemia were diagnosed. Candidemia with infection of the central venous catheter was the most frequent diagnosis (52%); in 20% of cases a tissue localization was also present (disseminated candidiasis). Thirty-four out of 50 episodes occurred in the Surgical Department. Among risk factors the most frequent resulted: prolonged antibiotic treatment (100%), intravascular catheter (86%), parenteral nutrition (74%), abdominal surgery (46%). C. albicans was identified more frequently than others Candida spp., resistance to fluconazole was detected in 20% of strains tested. 34/45 episodes of fungemia were treated with fluconazole, none reported side effects. In 5 cases fluconazole was discontinued for clinical failure. Clinical outcome in patients with Candida infection depends on other factors beside in vitro drug susceptibility tests. PMID- 12085715 TI - [Locoregional treatment of hepatocellular carcinoma: comparison between percutaneous alcohol administration and intraarterial chemoembolization]. AB - Between 1991 and 1998, 45 cirrhotic patients with hepatocellular carcinoma (1 or 2 lesions smaller than 5 cm) were treated either with percutaneous ethanol injection (26 patients) or with trans arterial chemoembolization (19 patients) in our Department. Percutaneous ethanol injection was performed on 37 nodules: mean diameter of 3.1 +/- 0.8 cm. Transarterial chemoembolization was performed on 27 nodules: mean diameter of 2.8 +/- 0.7 cm. Therapeutic success at first treatment was obtained in 81.5% of percutaneous ethanol injection treated lesions and in 60.0% of trans arterial chemoembolization treated lesions. Local recurrence after first treatment occurred in 7 of 22 percutaneous ethanol injection patients (31.8%) and 8 of 15 transarterial chemoembolization patients (53.3%). Distant hepatic recurrence after first treatment occurred in 16 of 26 (61.5%) alcolization treated patients and in 14 of 25 (56.0%) chemoembolization treated. Multivariate analysis selected therapeutic success and modality of treatment as the best predictor of better survival probability in all treated patients. Survival probability at 12/24/36/48 months was 96.1/80.8/61.6/42.3% in percutaneous ethanol injection group and 89.5/68.4/36.8/15.8% in transarterial chemoembolization group (p < 0.05 Log rank). Few data are available comparing the efficacy of percutaneous ethanol injection and transarterial chemoembolization in patients with hepatocellular carcinoma. Our data confirm that percutaneous ethanol injection is better than trans arterial chemoembolization in the treatment of patients with one or two lesions of hepatocellular carcinoma. PMID- 12085716 TI - [Psychotic disorders induced by interferon alfa]. AB - The treatment with interferon-alpha is contraindicated in patients with major depression (National Institutes of Health), but a considerable number of neuropsychiatric side effects is associated with it: personality changes, impaired cognitive, mania, psychosis. In the literature limited reports of interferon-alpha-induced psychotic disorder exist. Authors report on a 46-year old man diagnosed with alcohol induced psychotic disorder, who had been started to a treatment with Pegylated Interferon alfa-2b [Peg-Intron] and Ribavirin owing to a previous HCV-induced chronic hepatitis associated with a significant change in liver enzymes. During the first two months after the starting of therapy liver enzymes normalised, and modest side effects emerged, but the patient developed a psychotic reaction. Symptoms disappeared after discontinuation of interferon treatment. PMID- 12085717 TI - [Severe idiopathic hyponatremia caused by ADH inappropriate secretion in the elderly]. AB - We report on seven elderly patients with severe hyponatremia (plasma sodium < 125 meq/l). All were symptomatic for central nervous system disturbances, weakness, nausea/vomiting and met clinical and laboratory criteria for the diagnosis of inappropriate secretion of antidiuretic hormone (SIADH). Investigations performed to determine the etiology of the syndrome gave negative results, so that the form was considered to be idiopathic. TC scans or MNR showed brain atrophy and/or chronic ischemic lesions. According to the scant series found in the literature, we believe that aging itself may be a risk factor for SIADH. Lowering of the osmolal threshold and/or reduced receptors sensitivity to osmotic stimula, induced by ischemic changes in the hypothalamic region, can represent the underlying mechanisms. PMID- 12085718 TI - [Hyponatremia in clinical practice]. AB - Hyponatremia is defined as serum sodium level below 135 mEq/l; this electrolyte disorder can be associated with low, normal or high plasma tonicity. Hyponatremia with normal plasma osmolality, pseudohyponatremia, has little clinical value. Hyponatremia with increased plasma osmolality results from hyperglycemia or mannitol infusion. Patients with hyposmotic hyponatremia may be normovolemic, hypovolemic or hypervolemic; it is most important to know clinical history, physical examination that focuses on volume assessment and laboratory evaluation that includes urine osmolality and urine sodium concentration. Severe hyponatremia is associated with neurological complications and occasionally with mortality; for mild hyponatremia water restriction is usually sufficient, but in serious cases hypertonic saline infusion should be administered. Rapid correction of severe hyponatremia can cause brain demyelination; to prevent brain damage the rate of correction should be no more than 0.5 mEq/l/h (10-15 mEq/l/24 h). PMID- 12085719 TI - [Systemic lupus erythematosus between clinical practice and the laboratory: state of the art and new findings on anti-DNA autoantibodies]. AB - Identification of autoantibodies directed against nuclear antigens is a very important finding in the assessment of autoimmune rheumatic diseases. In particular, the anti-DNA autoantibodies have assumed a fundamental importance, both speculative and clinical, in the study of the systemic lupus erythemathous. The aim of the present review is to focalize on anti-DNA the mechanisms of both induction and production of anti-DNA autoantibodies, pathophysiologic and diagnostic and clinical aspects. For this purpose, forty years of studies on this topic have been reviewed. Aspects on different conformational shapes of double stranded DNA have been discussed such as related pathogenetic and diagnostic ones. Finally, the review has dealt with experimental therapies, focusing on both animal models and the most recent clinical trials according to Evidence Based Medicine. PMID- 12085720 TI - Periodontal disease and cardiovascular disease: epidemiology and possible mechanisms. AB - BACKGROUND: Many early epidemiologic studies reported an association between periodontal disease and cardiovascular disease. However, other studies found no association or nonsignificant trends. This report summarizes the evidence from epidemiologic studies and studies that focused on potential contributing mechanisms to provide a more complete picture of the association between periodontal and heart disease. TYPES OF STUDIES REVIEWED: The authors summarize the longitudinal studies reported to date, because they represent the highest level of evidence available regarding the connection between periodontal disease and heart disease. The authors also review many of the case-control and cross sectional studies published, as well as findings from clinical, animal and basic laboratory studies. RESULTS: The evidence suggests a moderate association--but not a causal relationship--between periodontal disease and heart disease. Results of some case-control studies indicate that subgingival periodontal pathogenic infection may be associated with myocardial infarction. Basic laboratory studies point to the biological plausibility of this association, since oral bacteria have been found in carotid atheromas and some oral bacteria may be associated with platelet aggregation, an event important for thrombosis. Animal studies have shown that atheroma formation can be enhanced by exposure to periodontal pathogens. CONCLUSIONS: The accumulation of epidemiologic, in vitro, clinical and animal evidence suggests that periodontal infection may be a contributing risk factor for heart disease. However, legitimate concerns have arisen about the nature of this relationship. These are early investigations. Since even a moderate risk contributed by periodontal disease to heart disease could contribute to significant morbidity and mortality, it is imperative that further studies be conducted to evaluate this relationship. One particularly important study to be carried out is the investigation of a possible clinically meaningful reduction in heart disease resulting from the prevention or treatment of periodontal disease. PMID- 12085721 TI - The relationship between oral conditions and ischemic stroke and peripheral vascular disease. AB - BACKGROUND: This article is a critical review of the epidemiologic studies linking or disassociating oral conditions with stroke and peripheral vascular disease, or PVD. It also describes possible causal and non-causal pathways for these associations. TYPES OF STUDIES REVIEWED: The author reviewed studies that evaluated the oral conditions of periodontal disease or tooth loss as risk factors for arterial disease and stroke. RESULTS: Seven of nine studies evaluating tooth loss and periodontal disease as risk factors for stroke or PVD showed some significant associations. The studies varied in the exposures and outcomes evaluated; therefore, the associations were not consistently replicated. It is unclear whether the associations found between these oral conditions and cardiovascular disease had any causal component. In the absence of any causal relationship, the associations may be explained by common risk factors. Alternatively, there may be a causal relationship that may be explained by one or more potential causal pathways. Further epidemiologic studies are needed, and the role of nutrition and other inflammatory mediators needs to be explored further in this context. CLINICAL IMPLICATIONS: It is recommended that clinicians continue to provide standard professional care, as well as nutritional counseling to help patients maintain a healthy diet after extractions. Clinicians are cautioned against suggesting extractions as a means of preventing cardiovascular disease. PMID- 12085722 TI - Does chronic periodontitis cause coronary heart disease? A review of the literature. AB - BACKGROUND: Chronic periodontitis, or CP, has been associated with coronary heart disease, or CHD. This article reviews the available prospective CP-CHD evidence. LITERATURE REVIEWED: In nine cohort studies, CP was associated with a 15 percent greater risk of developing CHD. Conclusions from individual studies depended on study characteristics. Summary risk estimates for studies controlling for smoking intensity (five of nine studies) or health awareness (two of nine studies) or studies with more than 600 CHD events (three of nine studies) suggest that CP is either not at all or weakly associated with CHD. Summary risk estimates for the studies that did not control for these factors or that examined an insufficient number of CHD events reported a weak increase in CHD risk associated with CP (20 percent greater). These data suggest that the CP-CHD associations observed in smaller studies are due to insufficient control for lifestyle differences. In addition, one cohort study reported that edentulous people had a CHD risk similar to that of people with CP. Therefore, the plausibility of dental infection elimination affecting CHD risk appears limited. SUMMARY: Current evidence supporting a causal CP-CHD link is weak. Rigorous methodological and analytical control of lifestyle factors such as smoking will be required to elucidate whether the CP-CHD disease association is either small or nonexistent. CLINICAL IMPLICATIONS: Cigarette smoking destroys both oral and systemic health. Because of this strong common causal factor, oral and systemic health are linked. Dentistry should continue to play an important role in implementing smoking prevention and cessation programs. PMID- 12085723 TI - Oral care for patients with cardiovascular disease and stroke. AB - BACKGROUND: The authors describe how oral disease, particularly periodontal disease, may place certain patients at increased risk of developing cardiovascular disease and stroke. RESULTS: Although the precise mechanisms of interaction are not clear, two biological mechanisms that may explain the relationship are presented. In addition, the authors describe the dental management issues that need to be understood in treating patients compromised by cardiovascular disease. CONCLUSIONS AND CLINICAL IMPLICATIONS: The patient with cardiovascular disease may present a challenge to the dental health care provider, depending on the degree of hemodynamic compromise and the stability of his or her condition. PMID- 12085724 TI - Taking oral health to heart: an overview. AB - BACKGROUND: This article is an introduction to the 2001 ADA symposium, "Taking Oral Health to Heart," which focused on the relationship between periodontal disease and cardiovascular disease. This overview describes the databases and terminology used in discussions of epidemiologic studies on this topic. CONCLUSIONS: In determining if a certain disease, medication or condition is a risk factor for another entity, one must carefully evaluate and analyze the data, weigh the various other contributing risk factors and then determine the mechanism of interaction if an association is established. CLINICAL IMPLICATIONS: This symposium provided practitioners with various concepts regarding the role of periodontal disease as a risk factor for cardiovascular disease, and the data presented represent the state of the art. PMID- 12085725 TI - Infections and their role in atherosclerotic vascular disease. AB - BACKGROUND: Complications of atherosclerosis are the leading cause of mortality in developed countries, and infections may play a role in the pathogenesis. Numerous studies have addressed this issue in the past decade. TYPES OF STUDIES REVIEWED: The author examined peer-reviewed studies and reviews on the role of microbes or infections in atherosclerosis, cardiovascular disease and cerebrovascular disease. He included selected articles on epidemiology, pathology, in vitro experiments, animal models and clinical studies. RESULTS: Cross-sectional and retrospective studies have shown an association between Chlamydia pneumoniae antibodies and cardiovascular disease, but prospective studies have not been as convincing. Studies on the association between cardiovascular disease and periodontal disease or loss of teeth have produced conflicting results. Cytomegalovirus infection is associated mainly with accelerated arteriosclerosis after cardiac transplantation. Infectious agents can induce biological mechanisms important for atherogenesis. Mice and rabbit studies have indicated that C. pneumoniae is capable of initiating or accelerating the progression of atherosclerosis. Limited studies on cytomegalovirus also suggest the ability to induce early changes of atherosclerosis in a rodent model. Preliminary clinical trials of treatment for C. pneumoniae infection suggest a possible short-term benefit, but larger randomized trials for longer periods are in progress. CONCLUSION AND CLINICAL IMPLICATIONS: Infectious agents may play an important role in atherogenesis, but currently the jury is not in. Further management of cardiovascular disease could change radically if this concept were proven. PMID- 12085726 TI - Parenting attitudes and marital intimacy: a longitudinal analysis. AB - This longitudinal study examined change in 97 couples' marital intimacy over the first 3 years after the birth of a child. Participants included both first-time and experienced parents. Regardless of parity, both wives and husbands, on average, showed linear declines in marital intimacy; however, significant variability in individual trajectories was found. The study evaluated the relation between parenting attitudes, measured independently for each partner and in terms of agreement between partners, and individual differences in the level and trajectory of marital intimacy. For both wives and husbands, higher perceived difficulty with parenting was related to lower initial levels of marital intimacy. Wives whose husbands held more traditional attitudes regarding child rearing and those whose beliefs about child rearing differed from the beliefs of their partners experienced steeper declines in intimacy over time. PMID- 12085727 TI - Development of the stepfamily events profile. AB - This project developed and examined the validity of a tailor-made life events checklist specific to the experiences of adolescents in stepfamilies (i.e., the Stepfamily Events Profile; STEP). The STEP was developed from information gathered in focus groups and interviews and includes positive and negative events that are meaningful to adolescents in stepfamilies. The STEP was given to 90 adolescents residing with their biological mother and stepfather. Negative events were significantly related to adolescent report of maladjustment, and positive events were significantly related to mother and adolescent report of internalizing problems, above and beyond the effects of several covariates. Also, the effects of negative and positive events were uniquely predictive of adolescent maladjustment. Events reflecting interactions within the adolescent's dyadic family relationships were most strongly predictive of adolescent maladjustment. PMID- 12085728 TI - Lesbian and bisexual mothers and nonmothers: demographics and the coming-out process. AB - In a large, national sample of 2,431 lesbians and bisexual women, those who had children before coming out, those who had children after coming out, and those who did not have children were compared on demographic factors and milestones in the coming-out process. Differences were found in race/ethnicity, age, prior marriage, income, religion, use of mental health counseling, and reported hate crimes. Results are also presented for lesbians and bisexual women of each ethnic/racial and age group. Controlling for age and income, lesbians and bisexual women who had children before coming out had reached developmental milestones in the coming-out process about 7-12 years later than women who had children after coming out and about 6-8 years later than nonmothers. PMID- 12085729 TI - Mothers' and children's expectations for adolescence: the impact of perceptions of an older sibling's experience. AB - The present study examined the role of perceived experience with an older child in mothers' expectations for subsequent children's adolescence as well as the impact of perceived experience with an older sibling on young adolescent children's expectations for their own adolescence. Participants were 305 sixth- and seventh-grade adolescents and 287 of their mothers. Participants were interviewed via telephone. For both mothers and children, the quality of experience with an older child/sibling was a better predictor of target-based expectations for the young adolescent than was the simple presence or absence of an older sibling. In some cases the link between quality of experience with an earlier born adolescent and mothers' expectations for the target child's adolescence was modified by the target child's temperament. In general, the results support a modeling hypothesis. PMID- 12085730 TI - Early mothering experience and personality change. AB - This study examined the context of the early mothering experience and how that experience was related to personality change in 48 members of a longitudinal sample of women from age 21 (in 1958 or 1960), before any of them were married, to age 27 (in 1963 or 1964). Early motherhood was analyzed in terms of (a) how positively the woman described her experience and (b) how willing and able she was to maintain the full-time commitment considered necessary in the early 1960s. Path analyses showed that these two dimensions had somewhat different interrelations with mother's personality, the marriage, and husband's personality and participation as a father. However, both dimensions were related to relative increase in ego-resiliency and to relative decrease in feelings of vulnerability between ages 21 and 27, even when family and work variables were taken into account. PMID- 12085731 TI - What predicts good relationships with parents in adolescence and partners in adult life: findings from the 1958 British birth cohort. AB - This study drew on data from the National Child Development Study to explore the role of father involvement and mother involvement at age 7 in father-child and mother-child relations at age 16, and the role of closeness to father and closeness to mother at age 16 in quality of relationship with partner at age 33. Closeness to mother was associated with closeness to father, intact family structure and academic motivation, and closeness to father with closeness to mother, early father involvement, less emotional and behavioral problems in adolescence, male gender and academic motivation. Closeness to father at age 16 was more strongly related to level of father involvement at age 7 for daughters than for sons and to closeness to mother for sons than for daughters. Marital adjustment at age 33 was related to good relationships with siblings, mother, and father at age 16; less current psychological distress; female gender; and educational attainment. PMID- 12085732 TI - The influence of adult attachment styles on the association between marital adjustment and depressive symptoms. AB - This study tested the hypothesis that attachment styles moderate the relationship between marital adjustment and depressive symptoms among husbands and wives. In a sample of 91 married couples, ratings of the anxious-ambivalent attachment style moderated the relationship between marital adjustment and depressive symptoms for both husbands and wives. Additionally, ratings of the secure attachment style moderated the relationship between marital adjustment and depressive symptoms for wives, with a trend for husbands. These findings suggest a relationship between insecurity and a predisposition to depressive symptoms in marital relationships. PMID- 12085733 TI - Neighborhood safety and social involvement: associations with parenting behaviors and depressive symptoms among African American and Euro-American mothers. AB - The relation between neighborhood characteristics and parenting and the mediating role of maternal depressive symptoms was examined among African American and Euro American mothers of kindergarten children. Mothers' ratings of neighborhood safety were related to disciplinary strategies for both African American and Euro American mothers but not to expressions of affection. Interviewers' ratings of safety were related to mothers' use of hostile socialization strategies. Both mothers' and interviewers' reports of safety were linked with maternal depressive symptoms. Depressive symptoms mediated the relation between neighborhood safety and inconsistent discipline, suggesting that the influence of safety on inconsistent discipline was due to its impact on maternal depression. Although there were similarities across ethnic groups, the relation between social involvement and mothers' withdrawal of interactions with their children differed across groups. PMID- 12085734 TI - Evaluating process in child and family interventions: aggression prevention as an example. AB - This article reports on 2 studies designed to develop and validate a set of measures for use in evaluating processes of child and family interventions. In Study 1 responses from 187 families attending an outpatient clinic for child behavior problems were factor analyzed to identify scales, consistent across sources: Alliance (Satisfactory Relationship with Interventionist and Program Satisfaction), Parenting Skill Attainment, Child Cooperation During Session, Child Prosocial Behavior, and Child Aggressive Behavior. Study 2 focused on patterns of scale scores among 78 families taking part in a 22-week preventive intervention designed to affect family relationships, parenting, and child antisocial and prosocial behaviors. The factor structure identified in Study 1 was replicated. Scale construct validity was demonstrated through across-source convergence, sensitivity to intervention change, and ability to discriminate individual differences. Path analysis validated the scales' utility in explaining key aspects of the intervention process. Implications for evaluating processes in family interventions are discussed. PMID- 12085735 TI - [Reflections on the Perruche judgement]. PMID- 12085736 TI - [Atrial septal defect calibration: comparison of methods using balloons and 3D transesophageal echocardiography]. AB - The atrial septal defects (ASDs) occlusion by cardiac catheterization is an alternative to the surgical technique. The use of stretching by balloon is considered as the gold-standard for the ASDs calibration. Three dimensional transesophageral echocardiography (3D-TEE) brings an imaging of the ASD surface and allows the measurement of its maximal diameter. With the goal of a percutaneous ASD occlusion, seventy patients had a simultaneous calibration of the ASDs by 3D-TEE and the balloon method. The mean maximal diameter measured by 3D-TEE was at 20 + 15 mm (range 10-28 mm) compared to the mean balloon-stretched diameter at 22 + 5 mm (range 9-31 mm). The correlation between both methods is good (y = 3.15 + 0.77x, r = 0.8, p < 0.0001). The mean difference between the diameters measured by both methods was 1.85 + 3.17 mm. The ASD visualized by 3D TEE was single in 54 cases and multiple in 16 cases. The correlation between both methods was excellent in those with a single ASD (y = 1.74 + 0.84x; r = 0.85. p < 0.0001) but it was poor in those with multiple ASD (y = 12.4 + 0.4x; r = 0.45). The calibration by the balloon method which tests the septum resistance is an additive method to 3D-TEE which measures the anatomic diameter of ASD. PMID- 12085737 TI - [3D-transthoracic echocardiography: a selection method prior to percutaneous closure of atrial septal defects]. AB - Prior to putting in place a percutaneous device, the assessment of the atrial septal defect anatomy is mandatory. The 3D transthoracic echocardiography is a non-invasive method bringing an imaging of the septal defect surface and its borders. Fifty-two patients ageing from 3 to 16 years old had a rotational 3D transthoracic echocardiography. Thirty-six (69%) were selected for a percutaneous closure (borders > 7 mm and a septal surface/atrial septal defect ratio > 2). Thirty-two of these selected patients (89%) benefited from the Amplatz prosthesis implantation with success. The maximal 3D diameter of the septal defect was 20 + 4 mm (14-30) compared to the mean size of prosthesis at 22 mm (18-30). Four of the 36 patients (aneurismal septum or a borderline septal surface/atrial septal defect ratio) were secondarily oriented to surgeons. Sixteen patients (31%) were selected directly to a surgical closure. The lack of borders or septal surface were confirmed by the surgical view. The 3D transthoracic echocardiography allows to define new criteria for the selection of patients prior to a percutaneous closure of atrial septal defects. This prospective study demonstrates that 9 out of 10 patients who had the 3D criteria had a percutaneous closure of the septal defect. PMID- 12085738 TI - [Criteria for percutaneous non-closure of interatrial communications of the ostium secundum type by the Amplatzer septal occluder]. AB - The aim of the study is to recognise the criteria preventing the percutaneous closure of an interatrial communication of the ostium secundum type for the Amplatzer septal occluder device. From January 1999 to December 2000, 121 consecutive patients with an average age of 24.8 +/- 19 years affected with an interatrial communication of the ostium secundum type underwent echocardiographic examination with transthoracic Doppler to evaluate the maximal diameter of the communication, the border dimensions, and the length of the interatrial septum; 92.5% of them underwent haemodynamic investigation to evaluate the shunt and the stretched diameter. Comparison of the umbrella group averages with the surgical group was performed by the Student test, and the frequencies by the Chi 2 test. ROC curves were drawn for the numeric parameters. After these 2 examinations, 68 patients underwent an attempt at percutaneous closure with 61 successful (84.7%) and 53 had a surgical closure straight off. The surgical group was younger, with a more significant shunt and a wider communication. The criteria for non-closure apart from abnormal pulmonary venous reflux were: insufficient border (59% of cases), diameter too wide (16%), multiperforate septum (16%) and insufficient septum length (9%). Percutaneous closure could be performed in 2/3 of interatrial communications without abnormal pulmonary venous reflux. Candidates for surgery were younger, with a more significant shunt and a wider diameter. In retrospect 8 patients of the surgical group could have had percutaneous closure and 7 patients of the umbrella group could have been sent straight for surgery. PMID- 12085739 TI - [Patent ductal arteriosus occlusion by Rashkind umbrella and by detachable coil]. AB - The objective of this study is to report all of our experience of patent ductus arteriosus closure by interventional catheterisation, comparing two systems used successively: Rashkind umbrella and detachable coil. METHOD: Between January 1991 and July 2001, 72 patients underwent cardiac catheterisation in order to occlude patent ductus arteriosus (PDA). In 3 patients closure was not attempted (n = 1) or aborted (n = 2). The 69 patients in whom one or several prostheses were implanted are the object of this retrospective study. The patients were between 10 months and 18 years old (median 4 years), weighing between 6.7 and 54 kg (median 17 kg). The narrowest average angiographic diameter of the PDA was 2.2 mm (1 to 4 mm), type A in the Krichenko classification in 59 cases. The PDA was occluded by an umbrella in 29 patients (group 1), and from 1997 coils were used in 40 patients (group II). The medium term results were evaluated clinically and by colour doppler echocardiography. RESULTS: The age, weight, size and type of PDA were similar in the 2 groups. Group 1: 28 patients were treated with a single umbrella and one patient received 2 umbrellas. Systemic embolisation occurred in one case. The average period of follow-up was from 4 years to 10 years (average 6.5 years). The rate of residual shunt at 24 h, 6 months, and 12 months was 43%, 43%, and 39% respectively. A second implant was necessary in 2 children after 7 months and 30 months. The spontaneous disappearance of residual shunt was observed in 8 patients after between 1 month and 54 months (average 33.5 months). A slight residual shunt persisted in 4 patients (13.7%), 4.5 to 8.5 years after placing one or two umbrellas. The patients with a residual shunt were younger: 37 months versus 73 months (p < 0.05). Group II: 34 patients received a single coil and 6 patients several coils. Two cases of embolic migration and two cases of haemolysis were observed. The follow-up extended from 4 months to 4.5 years (average 2.2 years). The rate of residual shunt at 24 h, 6 months, and 1 year respectively was 35%, 10.5%, and 3.3%. At 6 and 12 months this rate was significantly less in group I (p < 0.01). Implantation of supplementary coils was necessary in 2 children at 24 h and at 9 months. Spontaneous disappearance of residual shunt occurred in 10 children out of 14 with an average interval of 5.5 months. A weak residual shunt remained detectable in 2 patients (5%) at 6 months and 36 months. CONCLUSION: The rate of initial residual shunt is comparable using both techniques. Spontaneous disappearance of residual shunts was observed in the majority of cases, but with the detachable coils this outcome is faster and the final rate for residual shunt is very low. PMID- 12085740 TI - [Three-dimensional multislice CT scanner: value in patients with pulmonary atresia with septal defect]. AB - The aim of this study was to evaluate the interest of multislice CT scanning using three-dimensional reconstructions for pre- or postoperative management of patients with pulmonary atresia with ventricular septal defect. Twenty-one consecutive patients (median age: 8 months, median weight: 5.6 kg) with pulmonary atresia with ventricular septal defect were explored by multislice CT. Standard protocol consisted in one helical thoracic acquisition, associated with an injection of 2 to 3 mL/kg of iodinated contrast medium at a flow rate from 0.5 to 1 mL/s. Three-dimensionnal reconstructions were systematically performed. Before operation, status and size of central pulmonary arteries, major aorto-pulmonary collateral arteries, and associated anomalies were analyzed. In 13 operated patients, the permeability of palliative anastomosis and postoperative anatomical changes were studied. The status of central pulmonary arteries was assessed in all cases and the sizes found were in agreement to surgical findings. Main aorto pulmonary collateral arteries were depicted and their relative position to central airways was assessed. Various associated anomalies were found: right aortic arch, anomalous coronary artery, abnormal pulmonary venous return, left superior vena cava. The anastomosis and the surgical connections (unifocalization, complete repair) were clearly analyzed. The multislice CT with three-dimensional reconstructions is a complementary imaging technique, reliable, useful to pre-operative management and postoperative follow-up of patients with pulmonary atresia with ventricular septal defect. PMID- 12085741 TI - [Important of magnetic resonance angiography with gadolinium injection in pulmonary vein diseases]. AB - The study of the pulmonary veins by echocardiography is sometimes difficult especially when the ultrasonic window is restricted. Conventional angiography is the classic reference examination but it exposes the patient to ionising radiation and requires the injection of an iodine contrast product. Another technique that can provide the essential information is magnetic resonance angiography (MRA) with injection of gadolinium. It was performed in 9 patients for suspected congenital or acquired anomalies of the pulmonary veins between June 1999 and December 2001. The patient's ages varied from 1 month to 10 years. The examinations were carried out on a 1.5 T Vision machine (Siemens, Erlangen, Germany) without "cardiac gating" or apnoea after parental consent. MRA with gadolinium injection showed 5 drainage anomalies (3 partial pulmonary venous refluxes in the superior vena cava, 2 scimitar syndromes) and 3 stenoses (one due to compression by an aneurysm of the left pulmonary artery, a second secondary to pericardial agenesis, and a third secondary to hypoplasia of an isolated vein). MRA allowed three dimensional visualisation of these anomalies. This is a rapid, non-invasive and certain imaging technique which does not expose the patient to ionising radiation. It is therefore of significance in the investigation of anomalies of the pulmonary veins complementing echocardiography, and could in future replace cardiac catheterisation. PMID- 12085742 TI - [Homozygotous mutation of the SCN5A gene responsible for congenital long QT syndrome with 2/1 atrioventricular block]. AB - Long QT syndrome is characterized by a prolongation of the QT interval on the surface ECG. This clinically and genetically heterogeneous cardiac disease is potentially lethal due to ventricular polymorphic tachyarrhythmias leading to syncope or sudden death. It is transmitted according to different mendelian modes due to mutations in several genes coding for cardiac ion channels. Heterozygous mutations in KCNQ1, HERG, SCN5A, KCNE1 and KCNE2 genes are responsible for the dominant form without deafness whereas homozygous mutations in KCNQ1 and KCNE1 are responsible for the recessive form (Jervell and Lange-Nielsen syndrome) associated with congenital deafness. We report the case of a 5 year-old boy referred for syncope with a prolongation of the QTc interval (526 ms) and a 2/1 Atrio-Ventricular (AVB) block on the surface ECG. Under beta-blocking therapy, the sinus rate decreased and the 2/1 AVB disappeared. Electrophysiological study evidenced an infra-hisian block and a unipolar ventricular endocardial pacemaker was implanted. A V1777M missense mutation was identified in the C-terminal part of SCN5A, cardiac sodium channel gene, at the homozygous state in the proband and at the heterozygous state in both parents and 2 sibblings. Only the proband had a severe phenotype with syncope and AV conduction anomalies. All other genetically affected subjects were asymptomatic. This study provides evidence for the involvement of homozygous LQT3 forms in "functional" AVB. PMID- 12085743 TI - [Late atrial tachycardia after Fontan-type procedure. Cooperative study of 52 cases]. AB - The Fontan procedure has allowed to improve the outcome of complex congenital cardiopathy involving single ventricle. A better understanding of the systemic venous circulation has favored bicavo-bipulmonary derivation instead of atrio pulmonary derivation. However, in spite of the improvement in surgical procedures, post-operative arrhythmias still occur with an increasing rate during follow-up reaching 40 to 50% of the patients in some series. We report a series of 52 patients of which 92% presented a severe atrial arrhythmia (atrial fibrillation or atrial flutter) during a 6-year follow-up. The outcome was worse in case of classic or modified Fontan (n=15) or direct dicavo-bipulmonary procedures (n=7). The non-modified Fontan group was characterized by a lower functional class (63% NYHA class I or II), more refractory atrial arryhthmias (37%), more deaths or transplanted patients (26%). Amiodarone was very effective in this context as opposed to the failure of class 1 anti-arrhythmic drugs. However, low dosage amiodarone in combination with a beta-blocker is recommended taking into account the important rate of amiodarone-induced side effects (53%). Atrial arrhythmia ablation was unsuccessful (8/10 failures). Anti-arrhythmic surgery (N=3) has been incompletely evaluated. In summary, transformation of failing Fontan procedures into bicavo-bipulmonary derivations seems to offer the best outcome at the price of a high surgical risk. PMID- 12085744 TI - [Vagal hyperreactivity and sudden infant death. Study of 15 families]. AB - Prone sleep position is obviously the main risk factor for sudden infant death. Other risk factors, such as vagal overactivity particularly in the familial form, are still discussed. We here report 15 families characterized by the coexistence of vagal overactivity and sudden infant death. At least, 1 child for each family had documented [Holter or occulo-cardiac reflex (OCR)] vagal overactivity. In 5 families 2 children were affected; in 2 families 3 children were affected and in 1 family 4 children were affected. Sudden death occurred in the elderly of the family in 8 cases, in the twin in 3 cases, in the 2nd in 3 cases and in the 5th child in 1 case. Within the 15 families, at least 1 parent had experienced vagally-induced fainting or syncope in 10 cases. Familial pattern of vagal overactivity is underlined. Possible links between vagal overactivity, risk factor for suddden death and sudden death are discussed. We suggest an Holter-ECG and OCR follow-up for sudden infant death siblings with history of familial vagal overactivity (3 examinations during the 1st year of life, at 1, 3 and 9 months). PMID- 12085745 TI - [Value of fetal echocardiography in screening for congenital heart diseases. Report of 1,049 consecutive examinations]. AB - Specialised antenatal echocardiographic screening has allowed 1049 consecutive examinations in 992 patients resident in the Aquitaine region over a period of nine years. The most frequently encountered indications are expertise (suspicion of cardiopathy during initial screening) and family history. Expertise allows a higher detection rate (46.5%) compared to the other indications. We have screened 150 cardiopathies in utero and among them 143 were able to followed up after delivery. We have noticed a much greater severity of the anomalies compared to those encountered in paediatric cardiology. An associated chromosomal anomaly was noticed in 9% of cases and more frequently for atrio-ventricular communication and conotruncal cardiopathies. The rate of abortion was 35%, and the mortality for the surviving pregnancies was elevated (35%). Pharmacological treatment was instituted in 95% of cases of sustained supraventricular tachycardia with 81% successful. Diagnosis errors concerned benign cardiopathies above all and did not change the management of the pregnancy. The sensitivity of fetal echocardiography was 93.1% and its specificity 99.1%. This study shows the viability of fetal cardiac echography for the detection of the majority of congenital cardiopathies in a population at risk and in the management of fetal cardiac rhythm disorders. Nevertheless it remains a specialised examination for which the performance depends much on the initial screening performed by gynaecologists. Although the mortality rate of the screened children is elevated, indicating the frequent parental choice of abortion and the complexities of the cardiopathies, antenatal diagnosis has allowed optimisation of the neonatal management for certain curable cardiopathies. PMID- 12085746 TI - [Marfan syndrome in the newborn and infants less than 4 months: a series of 9 patients]. AB - We report a series of 9 children with neonatal Marfan syndrome. The diagnosis was made on a striking facial dysmorphia associated with arachnodactyly type skeletal anomalies. They all had cardiovascular anomalies: aortic dilatation 9 times, and mitral and/or tricuspid valve prolapse 8 times. Three children underwent ascending aorta replacement with the Yacoub technique. There was one operative death in a young infant and the two survivors required further surgery for gross mitral insufficiency and an aneurysm of the horizontal aorta. Three children died before the age of 1 year from cardiac insufficiency linked to massive leaks of all the valves. The three youngest children in the series did not undergo any procedure and were followed up for a slowly growing dilatation of the initial aorta. The prognosis of neonatal Marfan syndrome is weighted with a heavy mortality. The timing of surgical intervention in case of excessive dilatation of the aortic root is difficult to determine. It depends not only on the aortic diameter but also on the aortic valve function and the existence of other mitral or tricuspid lesions. Surgical treatment for lesions of the initial aorta and the atrioventricular valves improves the short-term prognosis but the risk of re intervention is higher. PMID- 12085748 TI - [Perforation of the aorta after percutaneous closure of an atrial septal defect with an Amplatz prosthesis, presenting with acute severe hemolysis]. AB - We report a case of aortic perforation three weeks after transcatheter occlusion of an atrial septal defect (ASD) by an Amplatzer device. Revealed by acute hemolysis, this complication needed an emergency surgical operation. The fistula between the no coronary Valsalva sinus of the aorta and the left atrium was repaired. The ASD was closed by patch. This serious accident should consider a short antero-superior rim as a risk factor for aortic perforation in transcatheter closure for ASD. PMID- 12085747 TI - [Outcome of infants with hypoplastic left heart syndrome diagnosed in utero]. AB - The authors report the results of prenatal diagnosis of the hypoplastic left heart syndrome since 1998 in the University Hospitals of Marseille and Montpellier. Twenty-four prenatal diagnoses of this condition were made in mothers with a mean age of 29 (18 to 40 years) and after a mean term of 22 (18.5 to 33) weeks of amenorrhea. Seventeen therapeutic abortions were carried out and 7 neonates born after a mean term of 39 (28 to 40) weeks, were admitted to the paediatric intensive care unit. Two patients required ventilatory assistance with one early death. The other patients were stable after surgery. A Norwood (first stage) procedure was carried out in 6 neonates at a mean age of 5 (1 to 6) days. There was only one survivor (17%). Prenatal diagnosis of the hypoplastic left heart syndrome allows cardiac and extracardiac evaluation of foetuses with this condition. Therapeutic abortions may be proposed and was the commonest choice of the parents in this study. On the other hand, despite better management of neonates with this prenatal diagnosis, the poor prognosis of the Norwood first stage procedure is unchanged. A systematic search for a restriction of the foramen ovale on foetal echocardiography could optimise neonatal management of this problem. PMID- 12085749 TI - [Percutaneous closure of a paravalvular mitral regurgitation with Amplatzer and coil prostheses]. AB - The authors report a case of percutaneous closure of paravalvular mitral regurgitation in a 9 year old child. This patient had complete atrioventricular canal with levo-isomerism completely repaired with replacement of the left atrioventricular valve by a mechanical prosthesis. One year after an episode of infectious endocarditis, the patient was readmitted because of jaundice due to haemolysis attributed to paravalvular mitral regurgitation. Transoesophageal echocardiography demonstrated a crescent-shaped dehiscence of the prosthesis opposite the left atrial appendage associated with a small paraseptal regurgitation. First of all, a 6 mm Amplatzer Septal Occluder and a 5 x 5 coil were introduced by retrograde catheterisation to treat the regurgitation due to the valve dehiscence. This procedure was completed 3 months later by the implantation of a second 8 mm Amplatzer prosthesis for persistent haemolysis. Closure of the paraseptal regurgitation resulted in a reduction of the patient's transfusional requirements. PMID- 12085750 TI - [Right atrial compressive aneurysm with favorable outcome after surgery at the age of 1 month]. AB - Right atrial aneurysms are uncommon and often revealed by arrhythmia. We report a case of right atrial aneurysm which required an early management and its follow up during one year. The abnormality was detected at 32 weeks gravida during a systematic echography. At birth the newborn was asymptomatic but with the progressive increase of the aneurysm size shifting the heart and vessels and the occurrence of an intra-cardiac thrombus, a surgical management was decided at the age of one month. The resection of the aneurysm was therefore performed and no complication occurred during recovery. After one year the child remains healthy. The surgical indications are difficult to precise as the majority of these aneurysms are detected at an adult age. Surgical management appears to be essential in case of poor tolerance. The prognosis after surgery seems favorable. PMID- 12085751 TI - [Placement of an implantable defibrillator via the endocavitary route in a patient presenting with a univentricular heart and a Glenn anastomosis]. AB - We report the observation of a female 45 year old patient presenting with a post surgery complex congenital cardiopathy, associated with serious ventricular rhythm disorders necessitating the placement of an implantable defibrillator. The palliative surgery performed (cavo-pulmonary Glenn anastomosis) does not allow the usual access to the right ventricle via the superior vena cava. The different possibilities for defibrillator implantation are discussed. These include associating a surgical approach to introduce the bipolar probe with subcutaneous tunnelling to connect the probe to the box. PMID- 12085752 TI - [Dextrocardia with situ solitus, ventricular loop, double outlet hypoplastic right ventricle and L-malposition of the great arteries. Description and surgical treatment of a rare and complex cardiopathy]. AB - Despite the cardiac surgery improvements allowing the correction of the majority of congenital heart diseases with ventricle-great vessels discontinuity, some abnormalities increase the risk of bi-ventricular reparation. We herein report the case of a patient presenting a rare form of double outlet right ventricle with a ventricular loop, with moderate right ventricle hypoplasia. L-malposition of great vessels and pulmonary artery stenosis, and for whom we opted for a palliative surgical treatment including a systemic-pulmonary anastomosis followed by a upper right bi-directional cavo-pulmonary derivation. The last surgery was followed by recurrent right pleural effusions disappearing after the embolization of the systemic-pulmonary anastomosis by catheterism as it probably obstructed the draining of the cavo-pulmonary anastomosis. The relevance of this clinical case reported is, firstly the description of this ventricle loop resulted from a marked ventricular malposition which is a rare heart disease, and secondly the discussion about the surgical treatment, especially about the choice between palliative and curative surgery. Only comparative studies on long term morbidity and mortality between the bi-ventricular reparation and mono-ventricular palliation will allow the selection of the most appropriate surgical treatment. PMID- 12085753 TI - [Interarterial and intramural coronary trajectoris: anatomic aspects and surgical implications]. AB - Among the coronary trajectory anomalies found not only in congenital cardiopathies but also in otherwise normal hearts, the terms intramural and interarterial are still used in an ambiguous fashion too frequently. Study of the embryonic development of the coronaries allows a better understanding of the possibility of such a trajectory, (existence of the periarterial sheath). Certain anatomical factors favour the existence of an intramural coronary trajectory, the coronary always being tangential to the aorta in this case: paracommissural ostium, ostium situated too high, departure of the coronary at an acute angle, and ostium in the form of a slit. However, interarterial and intramural are not synonyms. In the case of a coronary trajectory tangential to the aorta, three anatomical and histological situations are possible: free trajectory, with the aorta and the coronary each having its own media and its own adventitia; adherent trajectory, with two separate media covered by a common adventitia: and true intramural trajectory, with fusion of the media without interposition of adventitia. These three situations have different surgical implications in the case of coronary reposition. PMID- 12085754 TI - [Platypnoea-orthodeoxia syndrome. Diagnosis, etiology, treatment]. AB - Platypnoea-orthodeoxia (P.O.) syndrome is the association of dyspnoea and arterial oxygen desaturation aggravated in the erect position and relieved in the supine position. Initially considered very rare (20 cases reported over fifty years) and occurring essentially in patients having undergone pneumonectomy, it in fact occurs much more frequently if only it is looked for (20 extra cases reported in a single year). Some new aetiologies have been described, in particular dilatations or aneurysms of the ascending aorta. Diagnosis is made easier if the oxygen saturation is measured in the supine position when it is normal, and in the erect position when it falls considerably. Another argument is added by the small effect of inspiration of high concentrations of oxygen. Contrast echocardiography confirms the right-left shunt and allows estimation of the site. This is usually situated at the atrial level, via an inter-atrial communication or more often a patent foramen ovale; as a rule the area of dehiscence having been modified by the associated pathology allowing the right left shunt despite normal right pressures. Exceptionally the shunt is situated at the vascular or pulmonary parenchymal level. Poor tolerance of P.O. syndrome justifies a therapeutic procedure; this is usually closure of the septal fault with an occluder introduced percutaneously; the results are generally highly spectacular. PMID- 12085755 TI - [Ebstein anomaly. Anatomic aspects]. PMID- 12085757 TI - [Ebstein anomaly. Rhythm disorders and their treatment]. PMID- 12085756 TI - [Ebstein anomaly. Developmental aspects]. PMID- 12085758 TI - [Ebstein anomaly and pregnancy]. PMID- 12085759 TI - [Ebstein anomaly. Surgical treatment]. PMID- 12085760 TI - [Jawbone reduction in immediate prostheses. A comparison between overdentures and complete dentures]. AB - In a randomized controlled clinical trial 74 patients who required immediate dentures were randomly treated with immediate overdentures on 2 lower canines or immediate complete dentures. Mandibular bone reduction was measured using oblique lateral cephalometric radiographs made at base-line and the results compared with those of one year and two years after denture treatment. Analysis of the data showed that the average bone reduction in the lower canine regions in the first year was 0.75 in the immediate overdenture group and 1.90 mm in the immediate complete denture group. In the posterior parts of the mandible the bone reduction was respectively 0.77 and 1.77 mm. The differences were statistically significant in all measured regions. The differences in the first two years were significant in all regions except the molar region. Retention of roots of canines beneath a mandibular denture in immediate denture patients, even when they were in poor condition, reduced the collapse of the alveolar processes in all region of the mandible. PMID- 12085761 TI - [Reaction to, "The question of full extraction"]. PMID- 12085762 TI - [Prosthesis treatment of nursing home residents. New prosthesis preparation or improvement of existing one?]. AB - From a clinical point of view one should show restraint in preparing new dentures for elderly people. It is advised to improve existing dentures. This research project shows that both treatments in elderly, somatically handicapped, nursing home residents do not diminish their complaints and dissatisfaction. Obviously, they are best served by a problem-centred treatment. PMID- 12085764 TI - [Symposium: "The Dutch professional journal: a position localization"]. PMID- 12085765 TI - [Delightful counterfeits and monstrous mugs. Ideal and timeliness in the portrait art of the Renaissance]. PMID- 12085763 TI - [Jacob van der Haar and his therapy for "the swollen, enlarged and drooping tongue" as told to Camper(1782)]. PMID- 12085766 TI - [The sad decline of the ivory town. Toothache in letters and travel stories]. PMID- 12085767 TI - Screening for the target gene of cyanobacterial cAMP receptor protein SYCRP1. AB - The target genes for SYCRP1, a cyanobacterial cAMP receptor protein, were surveyed using a DNA microarray method. Total RNAs were extracted from a wild type strain and a sycrp1 disruptant of Synechocystis sp. PCC 6803, and the respective gene expression levels were compared. The expression levels of six genes (slr1667, slr1168, slr2015, slr2016, slr2017 and slr2018) were clearly decreased by the disruption of the sycrp1 gene. The data suggest that slr1667 and slr1668 constitute one operon and the other four genes constitute another operon. Transcription start points for the first genes of these putative operons, which are slr1667 and slr2015, were determined by primer extension experiments. Gel mobility shift assays and DNase 1 footprint analyses were carried out to explore the binding of SYCRP1 to the putative promoter regions of slr1667 and slr2015. SYCRP1 bound to the specific site in the 5' upstream region of slr1667 from positions -170 to -155 relative to the transcription start point, while it did not bind to the 5' upstream region of slr2015. It was concluded that SYCRP1 regulates the expression of the slr1667 gene directly by binding to a specific site in its promoter. PMID- 12085768 TI - One day makes a difference. PMID- 12085769 TI - Back from the brink. PMID- 12085770 TI - A moving experience. PMID- 12085771 TI - Listen up. PMID- 12085772 TI - Talking about bowels. PMID- 12085773 TI - Hungry for action. PMID- 12085774 TI - Medicines and breastfeeding. PMID- 12085775 TI - Nutritional advice in pregnancy. PMID- 12085776 TI - One-time screening for colorectal cancer with combined fecal occult-blood testing and examination of the distal colon. PMID- 12085777 TI - Endoscopic ligation compared with combined treatment with nadolol and isosorbide mononitrates to prevent recurrent variceal bleeding. PMID- 12085778 TI - Beating-heart bypass: pros and cons. PMID- 12085779 TI - I've always thought I was allergic to milk, but now I'm told that I have a food intolerance, not an allergy. What's the difference? PMID- 12085780 TI - A friend told me my heel pain is probably plantar fasciitis. What is this condition, and how is it treated? PMID- 12085781 TI - Protect your health in the air. Most people with chronic health conditions can travel by air safely--if they take a few precautions. PMID- 12085782 TI - Vanquishing varicose veins. Less invasive techniques work well and slash recovery time. PMID- 12085783 TI - Calcium's healthy cholesterol consequences. PMID- 12085784 TI - Migraine sufferers rarely see the doctor. PMID- 12085786 TI - New fix for farsightedness. PMID- 12085785 TI - Chicken may be kind to kidneys in people with type 2 diabetes. PMID- 12085787 TI - Childhood cancer survivors lack disease knowledge. PMID- 12085788 TI - Rheumatologists value alternative medicine. PMID- 12085789 TI - Reeling in more fish-oil findings. PMID- 12085790 TI - Hemorrhoid removal: stapler superior to scalpel. PMID- 12085792 TI - An unsweetened test for type 2 diabetes. PMID- 12085791 TI - (S)nifty treatment for pollen allergies. PMID- 12085793 TI - Does protein benefit bone density? PMID- 12085794 TI - A case in which the vagus influenced the form of the ventricular complex of the electrocardiogram. 1915. PMID- 12085795 TI - Regulation of intrinsic regenerative properties and axonal plasticity in cerebellar Purkinje cells. AB - Axon regeneration in the mammalian brain requires that injured neurons upregulate a specific set of growth-associated genes. To investigate the mechanisms that control the intrinsic growth properties of adult central neurons, we have examined the response to injury and regenerative potential of different cerebellar and precerebellar neuron populations. Axotomised neurons in the inferior olive, deep cerebellar nuclei and lateral reticular nucleus upregulate growth-associated molecules and regenerate their neurites into growth-permissive transplants. In contrast, Purkinje cells fail to respond to injury and show extremely poor regenerative capabilities. Targeted overexpression of GAP-43 promotes Purkinje axon plasticity, indicating that the weak regenerative potential of these neurons is mainly due to the inability to activate growth associated genes. Application of neutralising antibodies against the myelin associated protein Nogo-A induces cell body changes and axonal sprouting in intact Purkinje cells. In addition, immature injured Purkinje cells respond to axotomy and regenerate transected neurites, but they progressively lose this ability during postnatal development in parallel with myelin formation and the establishment of intracortical connections. These results indicate that the intrinsic growth potential of Purkinje cells is constitutively inhibited by environmental signals directed at stabilising the mature connectivity and preventing aberrant neuritic plasticity. Such a strict control eventually leads to restrict the regenerative capabilities of these neurons after injury. PMID- 12085797 TI - Colin Powell speaks frankly about HIV/AIDS. PMID- 12085796 TI - Forum brings updates of 9th Conference on Human Retroviruses. PMID- 12085798 TI - Summaries from the retrovirus conference, Part II. PMID- 12085799 TI - Victory for AIDS funding in Georgia. PMID- 12085800 TI - The role of civil society in South Africa 58th session of the Commission on Human Rights. PMID- 12085801 TI - National HIV Testing Day is June 27, 2002. "Take the test, take control!". PMID- 12085802 TI - Self-help by any name. PMID- 12085803 TI - Robots lend a helping hand to surgeons. PMID- 12085804 TI - Fighting gum disease: how to keep your teeth. PMID- 12085805 TI - Helping wounds heal. PMID- 12085806 TI - Itching for some allergy relief? PMID- 12085807 TI - First radiopharmaceutical for non-Hodgkin's lymphoma. PMID- 12085809 TI - HHS reaffirms value of mammogram screening. PMID- 12085808 TI - First HIV, hepatitis C screening system for blood donors approved. PMID- 12085810 TI - FDA continues work to help prevent mad cow disease. PMID- 12085811 TI - Maryland man, Virginia physician sentenced for illegally marketing aloe vera 'treatments'. PMID- 12085812 TI - The 'mouth-body connection': is there one? PMID- 12085813 TI - Kava and severe liver injury. PMID- 12085814 TI - More FDA resources to counter terrorism. PMID- 12085816 TI - FDA approves anthrax vaccine license supplements. PMID- 12085815 TI - Manufacturer discontinues only Lyme disease vaccine. PMID- 12085817 TI - FDA clears pocket-sized electrocardiograph. PMID- 12085818 TI - National Poison Control Hot Line launched. PMID- 12085819 TI - Pentobarbital in dog food. PMID- 12085820 TI - Study: majority of middle-aged Americans at risk of developing hypertension. PMID- 12085821 TI - Study shows St. John's wort ineffective for major depression. PMID- 12085822 TI - HHS, ADA work to boost awareness of diabetes and cardiovascular risk. PMID- 12085823 TI - Updated guidelines for managing HIV in pregnancy from the USPHS Task Force. PMID- 12085824 TI - Report from the 8th CROI: sensory neuropathy in HIV/AIDS. PMID- 12085825 TI - HPV-induced anal dysplasia: what do we know and what can we do about it? PMID- 12085826 TI - Addressing sexual risk behavior in HIV clinical practice. PMID- 12085827 TI - Update from the 8th CROI: adverse effects of HAART and structured treatment interruption. PMID- 12085830 TI - New study makes a compelling case for doubling defibrillator implants. PMID- 12085829 TI - And the survey says...client evaluation results. PMID- 12085828 TI - Helpline online forum. PMID- 12085831 TI - The pressure is still on to control hypertension. PMID- 12085832 TI - Bypass: restoring arterial flow is only the first step to recovery. PMID- 12085833 TI - Warning: these 8 heart myths could be hazardous to health. PMID- 12085834 TI - An infection-fighting drug appears to protect heart patients. PMID- 12085835 TI - New guidelines heighten safety for non-cardiac procedures. PMID- 12085836 TI - I have severe coronary artery disease and have been receiving chelation therapy. When I mentioned this to my doctor, he said I was wasting my money, and that I ought to stop. What do you think? PMID- 12085837 TI - I have tried everything I can think of to lose weight on my own and am now thinking of using weight-loss medications. Are they safe? PMID- 12085838 TI - During a routine physical, my doctor performed a chest x-ray. He said it showed I have an enlarged heart. What made him say this and should I be worried? PMID- 12085839 TI - Looking where the light is. PMID- 12085840 TI - The perils of interpretation. PMID- 12085841 TI - Stalking HIV immune dysregulation. PMID- 12085843 TI - AIDSWatch congressional lobbying, June 9-11 in Washington. PMID- 12085842 TI - Treatment access emergency: ADAP and Medicaid. PMID- 12085844 TI - Counterfit drugs: check Combivir, Serostim, Epogen. PMID- 12085845 TI - Medicaid funding cuts affect people with AIDS across the U.S. PMID- 12085846 TI - Johns Hopkins HIV treatment short guide. PMID- 12085847 TI - Remune controversy articles. PMID- 12085849 TI - Transplantation and HIV: facing two threats to life. PMID- 12085850 TI - Nine lives. PMID- 12085851 TI - The medicine of HIV-related organ transplantation. PMID- 12085852 TI - What you can do now to guard your heirs against heart disease. PMID- 12085853 TI - New device restores synchrony between ailing hearts' ventricles. PMID- 12085854 TI - Monitor all your medications to avoid drug-interaction risk. PMID- 12085855 TI - Snoring can be a wake-up call about cardiovascular diseases. PMID- 12085856 TI - I had bypass surgery performed nine years ago. Three grafts were placed from my leg veins. About two months ago, I developed the same pain that I had before my bypass surgery. Is this just in my head, or should I be worried? PMID- 12085857 TI - I recently received a letter from my cardiologist, and among other diagnoses listed was "sick sinus syndrome." What does this mean? PMID- 12085858 TI - I was experiencing shortness of breath and my doctor sent me to a cardiologist. He performed an echocardiogram. The diagnosis was "diastolic dysfunction." What's the significance. PMID- 12085859 TI - The effects of two cavity preparation methods on the longevity of glass ionomer cement restorations: an evaluation after 12 months. AB - BACKGROUND: The authors undertook a study to evaluate the effect of two cavity preparation methods on the initial survival of two more-viscous glass ionomer cements, or GICs, placed in the occlusal surfaces of permanent molar teeth. METHODS: Three dentists placed 149 restorations in 68 adult patients in a hospital clinic. They used either atraumatic restorative treatment, or ART, or conventional cavity preparation methods to place two encapsulated esthetic conventional GICs: Fuji IX GP (GC International Corp., Tokyo) and Ketac-Molar Aplicap (3M ESPE, Seefeld, Germany). For comparison, they used high-copper content GK Amalgam Alloy (Advanced Technology & Materials Co. Ltd., Beijing) in conventional preparations. They evaluated the restorations using both direct and indirect observation methods. RESULTS: Cavity preparations for which the authors used ART hand instruments took approximately twice as long to complete as did those for which they used conventional rotary instruments. After 12 months, no restorations had failed, but restorations comprising both GICs showed early losses of adjacent sealant material. Both GICs also showed relatively high restoration wear. At 12 months, the mean cumulative net occlusal wear for Fuji IX GP was 77 +/- 47 micrometers, and for Ketac-Molar 83 +/- 51 microm, without statistical significance (P > .05). Color matching improved significantly with time (P < .001), without significant differences in color between the two GICs by 12 months (P = .09). The amalgam alloy had minor surface tarnishing and marginal discrepancies increased with time (P < .00 1). CONCLUSIONS: All of the occlusal restorations were rated as satisfactory after 12 months. The method of cavity preparation did not affect the restoration performance of the GICs. However, their deterioration requires long-term monitoring. Clinical Implications. The more viscous GICs appear initially suitable for restricted use in clinical practice when placed using either of two cavity preparation methods in the occlusal surfaces of permanent molar teeth in adults. PMID- 12085860 TI - [Volumic and multislice CT: principles, applications and future prospects]. AB - The introduction of new array detector technology for multislice CT improves CT scan capabilities. This technique offers significant advantages including improved temporal and spatial resolutions, retrospective determination of slice thickness, shorter acquisition time, and contrast media saving. The most recent improvements lead to high quality cardiac and vascular examinations. The main principles of this technique are explained and its diagnostic as well as its economic impacts are discussed. PMID- 12085861 TI - Processing of animated contours by single cells in visual cortex area 18 of the cat. AB - In view of previous evidence that area 17 and 18 cells in cats represent the first level contour abstraction, we decided to study the specific response pattern of these cells to shapes enriched with contours, such as animated objects (AO). AO (cat's body whole contours) and 4 levels of AO degraded patterns were presented monocularly and repeatedly to the same eye, within the cells' RF. The intact AO elicited stronger responses than degraded contours while the responses to various degradedAO were similar. We conclude that under monocular presentation of AO cells react mainly as "edge detectors": the best stimulus is closed, compared with open, contour. PMID- 12085862 TI - Optic nerve compression by extramedullary hematopoietic tissue in a patient suffering from beta-thalassemia intermedia. AB - We carried out an encephalic magnetic resonance (MR) study in a patient with thalassemia intermedia who complained about change of the visual acuity and of the visual field. Narrowing of the optical canal was present and it was related to the compression of hemopoietic tissue masses. We found a clear relationship between the elderly age, the irregular transfusions, and the optic neuropathy. RMN screening of patients w ith thalassemia intermedia should be mandatory if changes in visual function occur. PMID- 12085863 TI - Treatment of scleral melt associated with uveal prolapse by pericardium grafting in Wegener's granulomatosis. AB - PURPOSE: To report the use of preserved pericardium tissue in the treatment of scleral melt associated with uveal prolapse secondary to Wegener's granulomatosis. Wegener's granulomatosis may initially present with ophthalmic findings including scleral melting with uveal prolapse. In order to preserve the globe and treat this potentially fatal disease, prompt diagnosis, surgical correction of uveal prolapse, and subsequent immunosuppressive therapy are imperative. If advanced scleral melt associated with uveal prolapse is present, pericardium grafting with conjunctival advancement is a useful technique. METHODS: Case report of a 70-year-old white female who initially presented with a unilateral sderal melt associated with uveal prolapse involving 6 clock hours of the surgical limbus. Emergent pericardium grafting of the involved limbus with conjunctival advancement performed to correct the endangered globe. Laboratory data including sedimentation rate, cANCA, computerized tomography of the chest, and urinalysis confirmed the diagnosis of Wegener's granutomatosis. RESULTS: After pericardium grafting and immunosuppressive therapy the patient returned to baseline visual acuity, the sedimentation rate normalized, and the pulmonary lesions and renal failure resolved. The patient has been followed for 3 years since the initial diagnosis of Wegener's granulomatosis and there has been no recurrence. CONCLUSIONS: To the best of the author's knowledge, this is the first report using processed pericardium to emergently treat scleral melt associated with uveal prolapse, thereby allowing preservation of the globe. In addition to management of the scleral melt, systemic evaluation and treatment with immunosuppressive therapy are necessary. PMID- 12085864 TI - Navy issues research guidelines. PMID- 12085865 TI - Legal implications of sterilization of the mentally retarded. AB - For more than 50 years, sterilization of mentally retarded persons was an accepted practice in many states as a way to reduce the number of persons born with mental defects. In recent years, the practice has come under heavy challenge, both medically and legally. At present, only 19 states have statutes that allow sterilizations for eugenic reasons. This article examines important court decisions regarding eugenic sterilization laws and the legal status of sterilization of mentally retarded adults and minors, as well as the consequent legal implications for physicians. Federal funding restrictions on sterilization of mentally impaired persons are discussed. The requirement of voluntary, informed consent is considered in light of an increasing emphasis on the rights of mentally retarded persons as well as greater awareness of the psychological effects of involuntary sterilization. PMID- 12085866 TI - Legislative threat to fetal research. PMID- 12085867 TI - Ethics and editors. PMID- 12085868 TI - Sickle cell "nondisease": a potentially serious public health problem. PMID- 12085869 TI - The role of ethics in pediatrics. AB - Medical ethics has increased in importance in medical education and practice as a consequence of advances in definitive treatment of patients. Most problems in ethics related to medicine have certain common denominators. Medical scientists and practitioners frequently disclaim any ethical absolutes or consider as important only nonmoral consequences rather than ethical principles such as justice, noninjury, truth-telling, etc. Failure to differentiate moral from nonmoral decisions as well as differences in level of moral reasoning also account for substantial differences in analyzing a given clinical problem. Finally, individual moral policy may be rendered ineffective because of institutional policies that result from purely pragmatic considerations. PMID- 12085870 TI - A considered approach to sterilization of mentally retarded youth. PMID- 12085871 TI - Ethics and editors. PMID- 12085872 TI - Clinical research and its ethical control in Durham between 1974 and 1979. PMID- 12085873 TI - Medical, moral and legal aspects of renal replacement therapy. PMID- 12085874 TI - From arguments to Supreme Court opinions in Planned Parenthood v. Casey. PMID- 12085875 TI - On euthanasia and assistance for a good death: plan for action. PMID- 12085876 TI - Application of Chakrabarty. 2 Mar 1978. PMID- 12085877 TI - Power of court to order or authorize discontinuation of extraordinary medical means of sustaining human life. PMID- 12085878 TI - Woman's right to have abortion without consent of, or against objections of, child's father. PMID- 12085879 TI - Tests of death for organ transplant purposes. PMID- 12085880 TI - Liability of one treating mentally afflicted patient for failure to warn or protect third persons threatened by patient. PMID- 12085881 TI - The philosopher as insider and outsider: how to advise, compromise, and criticize. PMID- 12085882 TI - Freedom, power, and physician-assisted death. PMID- 12085883 TI - Strength in numbers: the formation of the Metropolitan New York Ethics Committee Network. PMID- 12085884 TI - The patient's right to know: just who do the rules protect? PMID- 12085885 TI - Better off dead? PMID- 12085886 TI - Ethics and drug company gifts to physicians. PMID- 12085887 TI - The Kennedy Institute of Ethics. PMID- 12085888 TI - Genetic tests: safe enough? PMID- 12085889 TI - Test-tube babies: coming soon. PMID- 12085891 TI - Protection of human subjects. Effective 1 Sep 1975. PMID- 12085890 TI - In re Jane Doe. PMID- 12085892 TI - Right to natural death; brain death. PMID- 12085893 TI - When physician may treat minor without consent of parent, guardian or person in loco parentis. PMID- 12085894 TI - Communications between physician and patient. PMID- 12085895 TI - Sterilization of persons mentally ill and mentally retarded. PMID- 12085897 TI - Council on Sickle Cell Syndrome. PMID- 12085896 TI - Right to treatment. PMID- 12085898 TI - Informed consent to health care treatment or procedure. PMID- 12085899 TI - Act on establishment of an ethical council and regulation of certain forms of biomedical experiments. Act. No. 353. PMID- 12085900 TI - Patients, pills, and professionals: the ethics of placebo therapy. PMID- 12085901 TI - The allocation of limited medical resources: an ethical perspective. PMID- 12085902 TI - Pharmaceuticals for developing countries: an interface of science, technology, and public policy. PMID- 12085903 TI - The Oath of Asaph -- a structural analysis of the literature and of the biblical sources of its ethics. PMID- 12085905 TI - A paradigm of ethics for the maternal child nurse. PMID- 12085907 TI - The lone clone. PMID- 12085906 TI - Psychiatry, sociology and civil liberties. PMID- 12085908 TI - Issues in behavior control: an exchange of views. I-II. PMID- 12085909 TI - Genes, behavior, and what will become of us. PMID- 12085910 TI - Torture and the ethics of medicine. PMID- 12085911 TI - Guide to decision-making in institutional review of proposals for human experimentation. PMID- 12085912 TI - Induced abortion, Jewish law and Jewish morality. PMID- 12085914 TI - Human experimentation: issues before the hospital administrator. PMID- 12085913 TI - Ethics and hospital administration. PMID- 12085915 TI - The duties of physicians and some original results. PMID- 12085916 TI - An act...to establish a program of National Research Service Awards...and to provide for the protection of human subjects involved in biomedical and behavioral research.... Approved 12 Jul 1974. PMID- 12085917 TI - Nurses and the death penalty. PMID- 12085918 TI - Medicaid not required to fund medically necessary abortions. PMID- 12085919 TI - Pap smears: physicians must disclose risk of refusal. PMID- 12085920 TI - Infanticide and geronticide. PMID- 12085921 TI - Scientific procedures on living animals: proposals for reform of the 1876 Cruelty to Animals Act. PMID- 12085922 TI - On the re-use of laboratory animals. PMID- 12085923 TI - The biblical-Talmudic secret for choosing one's baby's sex. PMID- 12085924 TI - Bibliography of medical halacha. PMID- 12085925 TI - Animals in the research laboratory: science or pseudoscience? PMID- 12085926 TI - Moral standing, the value of lives, and speciesism. PMID- 12085927 TI - Dr. Frankenstein and today's professional biotechnologist: a failed analogy? PMID- 12085928 TI - Is the conduct of medical research on chimpanzees compatible with their rights as a near-human species? PMID- 12085929 TI - Animal experimentation and the argument from limited resources. PMID- 12085930 TI - Beyond Baby Doe: does infant transplantation justify euthanasia? PMID- 12085931 TI - The effect of microgravity and space flight on the chemical senses. AB - The effect of space flight and microgravity on the chemical senses is reviewed. Skylab-4 and Soyuz 30-31 studies revealed changes in taste thresholds while no effect was found in a Canadian investigation (41-G) and conflicting results were obtained on another Soyuz mission. Two simulated microgravity studies found no effect on taste or smell sensitivity; while 5 other studies found an effect. Microgravity induces physiological changes including an upward shift of body fluids toward the head, which may lead to an attenuation of the olfactory component in the flavor of foods. Chemosensory changes may also relate to space sickness, Shuttle atmosphere, stress, radiation, and psychological factors. PMID- 12085932 TI - Influence of modified starches on the stability of beef jerky analogs during storage. AB - Textural stabilization of extruded beef jerky analogs (BJA) by using modified starches was studied. From a series of modified starches studied, Frigex W and Instant Pure Flo were identified to show very reduced retrogradation, both in gels and after extrusion. These starches were used as textural stabilizers for BJA, an extruded product obtained from potato granules and ground beef. The partial substitution of potato granules with modified starches, at a level of 5% of the formulation, led to increased moisture sorption capacity of the analogs. After a storage period of 1 mo, the samples that contained modified starch had a degree of swelling with 30% higher and much lower elastic modulus as compared to those that contained only potato starch. PMID- 12085934 TI - Stem cells: equity or ownership? PMID- 12085933 TI - Numerical analysis of blood flow through a stenosed artery using a coupled, multiscale simulation method. AB - A global system model of the systemic circulation is combined with a local finite element solution to simulate blood flow in a stenosed coronary artery. Local fluid dynamic issues arise in connection with the detailed flow patterns within the stenosed coronary artery while the global system model is used to simulate the response of the rest of the circulation to the local perturbation. A PISO type finite element technique is employed to compute the local blood flow. The Navier-Stokes equations are solved with the assumption of viscous incompressible flow across the stenosed coronary artery. A detailed lumped parameter model simulates the characteristics of the coronary circulation and is imbedded in a coarse-grained lumped parameter model of the entire cardiovascular system. These two methods are coupled in that the lumped parameter calculations provide the time-dependent boundary conditions for the local finite element calculation. In turn, the local fluid dynamical computation provides estimates for the pressure drop across the stenosis, which is subsequently used to refine the lumped parameter calculation. Results are obtained for an axisymmetric coronary artery model with a stenosis of 90% area reduction over one cardiac cycle. Numerical results show that the flow rate and resistance are strongly coupled. Compared with the flow rate distribution computed from the global simulation with constant resistance, the coupled solution predicts a flow rate with only slight changes. The high flow rate during diastole increases the stenosis pressure drop and resistance. In turn, this increased resistance of the stenosis slightly reduces the flow rate computed in the lumped parameter simulation. PMID- 12085935 TI - Stem cell research: a target article collection: Part II--what's in a name: embryos, clones, and stem cells. AB - In 2001, the U.S. House of Representatives passed the "Human Cloning Prohibition Act" and President Bush announced his decision to allow only limited research on existing stem cell lines but not on "embryos." In contrast, the U.K. has explicitly authorized "therapeutic cloning." Much more will be said about bioethical, legal, and social implications, but subtleties of the science and careful definitions of terms have received much less consideration. Legislators and reporters struggle to discuss "cloning," "pluripotency," "stem cells," and "embryos," and whether "adult" are preferable to "embryonic" stem cells as research subjects. They profess to abhor "copying humans" or "killing embryos." Do they know what they are talking about? Do we? This paper explores the historical, philosophical, and scientific contexts that inform this heated discussion. PMID- 12085936 TI - Stem cell research: a target article collection: Part III--determining moral status. AB - In this chapter, I review some of the background thinking concerning matters of moral status that I had developed in previous years and that I would now bring to the work of the Human Embryo Research Panel. Two ideas were at the forefront of my thinking. First, that biology usually offers not decisive "events" but only continuous processes of development. Second, in making status determinations we do not so much "identify" a point on a developmental continuum where moral respect should be accorded as "choose" that point. These choices are "balancing decisions" in which the community of moral agents weighs its interests in protecting an entity against the burdens of doing so. After illustrating these two contentions, I consider some of the reasons why thinkers on the "right" and "left" of our bioethics debates have resisted or missed this basic insight. PMID- 12085937 TI - Stem cell research: a target article collection: Part I--Jordan's banks, a view from the first years of human embryonic stem cell research. AB - This essay will address the ethical issues that have emerged in the first considerations of the newly emerging stem cell technology. Many of us in the field of bioethics were deliberating related issues as we first learned of the new science and confronted the ethical issues it raised. In this essay, I will draw on the work of colleagues who were asked to reflect on the early stages of the research (members of the IRBs, the Geron Ethicist Advisory Board, and the National Bioethics Advisory Commission) as the field debated the issues of consent, moral status, use of animal tissues, abortion, use of fetal tissue, and the nature and goals of entrepreneurial research. In this new capacity, ethicists weighed the problem of privacy, the role of justice considerations, and the issues of the marketplace in science. At this point, it is clear that far more issues remain unresolved than are settled, that there is largely unexplored territory ahead, and that the single most important task that faces us as a field is a steady call for ongoing conversation and public debate. PMID- 12085938 TI - Ditching religion and reality. PMID- 12085939 TI - Crossing the ethical chasm: embryo status and moral complicity. PMID- 12085940 TI - Federal policy toward human embryonic stem cell research. PMID- 12085941 TI - Stem cell politics: the new shape to the road ahead. PMID- 12085942 TI - Stem cells and the man on the moon: should we go there from here? PMID- 12085943 TI - Those divisive stem cells: dealing with our most contentious issues. PMID- 12085944 TI - Embryonic stem cells: expanding the analysis. PMID- 12085945 TI - Embryonic stem cell retrieval and a possible ethical bypass. PMID- 12085946 TI - Stem cell research and the role of the new President's Council on Bioethics. PMID- 12085947 TI - Bush and stem cell research: an ethically confused policy. PMID- 12085948 TI - Ethics and politics. PMID- 12085949 TI - Stem cells, cloning, and abortion: making careful distinctions. PMID- 12085950 TI - Can moral worthiness be seen using a microscope? PMID- 12085951 TI - Questions concerning the current stem cell debate. PMID- 12085952 TI - Stem cell rhetoric and the pragmatics of naming. PMID- 12085953 TI - The idolatry of absolutizing in the stem cell debate. PMID- 12085954 TI - Stem cell ethics and the forgotten corporate context. PMID- 12085955 TI - Embryos, stem cells, and the "strategic" element of public moral reasoning. PMID- 12085956 TI - The moral status of the embryonal stem cell: inherent or imputed? PMID- 12085957 TI - Bioethics after the terror. AB - Bioethics as a field has been fortunate that its values and concerns have mirrored the values and concerns of society. In light of the September 11th attacks, it is possible that we are witnessing the beginning of a transition in American culture, one fraught with implications for bioethics. The emphasis on autonomy and individual rights may come to be tempered by greater concern over the collective good. Increased emphasis on solidarity over autonomy could greatly alter public response to research abuses aimed at defense from bioterrorism, to privacy of genetic information, and to control of private medical resources to protect the public health. PMID- 12085959 TI - Prostate cancer susceptibility genes: many studies, many results, no answers. AB - Since the first report of a genome-wide scan for hereditary prostate cancer (HPCA hereinafter) in 1996, several publications have presented data implicating various chromosomal regions by linkage analysis without any consequential identifications of the target genes. The most intensive attention has been focused on chromosome 1, and it has been proposed to contain at least three sub chromosomal regions (HPC1, PCAP, CAPB) harboring putative prostate cancer susceptibility genes. Nevertheless, one susceptibility gene, ELAC2/HPC2 at chromosome 17, has now been identified. Yet it seems to have a questionable role in prostate cancer predisposition. HPCA susceptibility loci have become undeniable archenemies of prostate cancer investigators, as the results of candidate gene analyses have been bewilderingly inconclusive. Predisposition to prostate cancer is most likely to be caused by several genes, different models of Mendelian inheritance, incomplete penetrance and varying population ethnicity frequencies. We will review the current state of the HPCA field and discuss the difficulties associated with identifying prostate cancer susceptibility genes. PMID- 12085960 TI - Molecular profiling in prostate cancer. AB - Prostate cancer is the most diagnosed cancer and the second leading cause of cancer death among men in the United States. Ability to detect this cancer early and availability of better prognostic markers are critical in order to decrease morbidity and mortality of prostate cancer. With the recent development in gene expression analysis methodology, expression profiles of thousands of genes can be generated in tissue samples and cell lines. Comparison of the global gene expression patterns between normal prostate and tumors at different stages may allow us to understand better the molecular mechanism of prostate tumorigenesis and progression. Different cancer cell lines and tissues appear to have different gene expression patterns that provide a new tool to classify tumors. Molecular classification of prostate cancer holds great promise for early detection and prognosis of this disease in the future. In this review, we summarize some of the recent mRNA and protein expression profiling studies performed in prostate cancer. Further, we discuss the potential benefits and limitations of current profiling technology. PMID- 12085961 TI - Chromosomal deletions and tumor suppressor genes in prostate cancer. AB - Chromosomal deletion appears to be the earliest as well as the most frequent somatic genetic alteration during carcinogenesis. It inactivates a tumor suppressor gene in three ways, that is, revealing a gene mutation through loss of heterozygosity as proposed in the two-hit theory, inducing haploinsufficiency through quantitative hemizygous deletion and associated loss of expression, and truncating a genome by homozygous deletion. Whereas the two-hit theory has guided the isolation of many tumor suppressor genes, the haploinsufficiency hypothesis seems to be also useful in identifying target genes of chromosomal deletions, especially for the deletions detected by comparative genomic hybridization (CGH). At present, a number of chromosomal regions have been identified for their frequent deletions in prostate cancer, including 2q13-q33, 5q14-q23, 6q16-q22, 7q22-q32, 8p21-p22, 9p21-p22, 10q23-q24, 12p12-13, 13q14-q21, 16q22-24, and 18q21 q24. Strong candidate genes have been identified for some of these regions, including NKX3.1 from 8p21, PTEN from 10q23, p27/Kip1 from 12p13, and KLF5 from 13q21. In addition to their location in a region with frequent deletion, there are functional and/or genetic evidence supporting the candidacy of these genes. Thus far PTEN is the most frequently mutated gene in prostate cancer, and KLF5 showed the most frequent hemizygous deletion and loss of expression. A tumor suppressor role has been demonstrated for NKX3.1, PTEN, and p27/Kip1 in knockout mice models. Such genes are important targets of investigation for the development of biomarkers and therapeutic regimens. PMID- 12085962 TI - Role of eicosanoids in prostate cancer progression. AB - Metabolism of arachidonic acid through cyclooxygenase, lipoxygenase, or P450 epoxygenase pathways leads to the formation of various bioactive eicosanoids. In this review, we discuss alterations in expression pattern of eicosanoid generating enzymes found during prostate tumor progression and expound upon their involvement in tumor cell proliferation, apoptosis, motility, and tumor angiogenesis. The expression of cyclooxygenase-2, 12-lipoxygenase, and 15 lipoxygenase-1 are up-regulated during prostate cancer progression. It has been demonstrated that inhibitors of cyclooxygenase-2, 5-lipoxygenase and 12 lipoxygenase cause tumor cell apoptosis, reduce tumor cell motility and invasiveness, or decrease tumor angiogenesis and growth. The eicosanoid product of 12-lipoxygenase, 12(S)-hydroeicosatetraenoic acid, is found to activate Erkl/2 kinases in LNCaP cells and PKCalpha in rat prostate AT2.1 tumor cells. Overexpression of 12-lipoxygenase and 15-lipoxygenase-1 in prostate cancer cells stimulate prostate tumor angiogenesis and growth, suggesting a facilitative role for 12-lipoxygenase and 15-lipoxygenase-1 in prostate tumor progression. The expression of 15-lipoxygenase-2 is found frequently to be lost during the initiation and progression of prostate tumors. 15(S)-hydroxyeicosatetraenoic acid, the product of 15-lipoxygenase-2, inhibits proliferation and causes apoptosis in human prostate cancer cells, suggesting an inhibitory role for 15 lipoxygenase-2 in prostate tumor progression. The regulation of prostate cancer progression by eicosanoids, in either positive or negative ways, provides an exciting possibility for management of this disease. PMID- 12085963 TI - Contribution of the androgen receptor to prostate cancer predisposition and progression. AB - Although prostate cancer is heterogeneous in its etiology and progression, androgen signaling through the androgen receptor (AR) appears to be involved in all aspects of the disease, from initiation to development of treatment resistance. Lifetime exposure to a constitutively more active AR, encoded by AR alleles as defined by two translated polymorphic microsatellites (CAG and GGC), results in a significant increase in prostate cancer risk. The AR gene is amplified or a target for somatic gain-of-function mutations in metastatic prostate cancer. Gain-of-function AR gene mutations may result in inappropriate activation of the AR, thereby contributing to the failure of conventional androgen-ablation treatments. In cases where no genetically altered receptors are observed, altered signaling through the AR, achieved by cross-talk with other signaling pathways (e.g. kinase-mediated pathways) and/or inappropriate expression of coregulatory proteins, may contribute to disease progression. Thus, the AR-signaling axis contributes to many aspects of prostate cancer, including initiation, progression and resistance to current forms of therapy. This recognition represents a paradigm shift in our understanding of the molecular mechanisms involved in progression of prostate cancer, and provides insight into novel AR-targeted therapies which ultimately may be more effective than current forms of androgen ablation. PMID- 12085964 TI - Regulation of apoptosis in prostate cancer. AB - Transformation and malignant progression of prostate cancer is regulated by the inability of prostatic epithelial cells to undergo apoptosis rather than by increased cell proliferation. The basic apoptotic machinery of most prostate cancer cells is intact and the inability to undergo apoptosis is due to molecular alterations that result in failure to initiate or execute apoptotic pathways. This review discusses the role of anti-apoptotic proteins such as Bcl-2/BclXL, NF kappaB, IGF, caveolin, and Akt, and pro-apoptotic molecules such as PTEN, p53, Bin1, TGF-beta, and Par-4 that can regulate progression of prostate cancer. In addition to highlighting the salient features of these molecules and their relevance in apoptosis, this review provides an appraisal of their therapeutic potential in prostate cancer. Molecular targeting of these proteins and/or their innate pro- or anti-apoptotic pathways, either singly or in combination, may be explored in conjunction with conventional and currently available experimental strategies for the treatment of both hormone-sensitive and hormone-resistant prostate cancer. PMID- 12085965 TI - On the role of cell surface carbohydrates and their binding proteins (lectins) in tumor metastasis. AB - This review focuses on the recent advances in investigations of the role of cell surface carbohydrates in tumor metastasis. It also summarizes the results of extensive studies of endogenous lectins, their structure, carbohydrate specificity and biological functions with the major emphasis on the significance of lectin-cell surface carbohydrate interactions in a metastatic process. Numerous data demonstrate that malignant transformation is associated with various and complex alterations in the glycosylation process. Some of these changes might provide a selective advantage for tumor cells during their progression to more invasive and metastatic phenotype. Cell glycosylation depends on the expression and function of various glycosyltransferases and glycosidases. Recently, transfection of genes encoding various glysosyltransferases gene in sense and antisense orientation helped to bring direct evidence that changes in cell surface carbohydrates are important for the metastatic behavior of tumor cells. Cell surface carbohydrates affect tumor cell interactions with normal cells or with the extracellular matrix during metastatic spread and growth. These interactions can be mediated via tumor cell carbohydrates and their binding proteins known as endogenous lectins. The family of the discovered endogenous lectins is rapidly expanding. The number of C-type lectins has reached 50 and at least 10 galectins have been identified. The biological significance of the endogenous lectins and their possible role in tumor growth and metastasis formation has started to unravel. Some lectins recognize the 'foreign' patterns of cell surface carbohydrates expressed by microorganisms and tumor cells, and play a role in innate and adaptive immunity. It was shown that lectins affect tumor cell survival, adhesion to the endothelium or extracellular matrix, as well as tumor vascularization and other processes that are crucial for metastatic spread and growth. PMID- 12085966 TI - Metastasis suppression in prostate cancer. AB - Due to a lack of effective treatments, the development of metastases remains the most lethal aspect of prostate cancer. In order to help overcome this problem there has been an ongoing effort to develop strategies for early intervention. This includes the development of strategies that allow histologic lesions and disseminated cells that are highly likely to cause metastatic disease to be distinguished from those that are not, as well as therapeutic approaches to specifically target bone metastases. Such approaches will be expedited by the identification of genes and signaling cascades that regulate metastatic growth. Genes that specifically suppress metastasis are strong candidates for these studies. This review will focus on metastasis-suppressor genes that have been identified functionally, particularly those found to play a role in prostate cancer, and discuss how the identification and study of these genes has influenced our overall understanding of the metastatic process. PMID- 12085967 TI - The urokinase-type plasminogen activator system in prostate cancer metastasis. AB - Accumulated clinical and experimental evidence indicates that the urokinase-type plasminogen activator (uPA) and its regulators are causatively involved in the metastatic phenotype of many types of cancers. In the past couple of decades, investigation on the role of the uPA system in human prostate cancer (PC) has been intensified and has yielded valuable insights. This review summarizes recent advances made in several areas regarding the clinical relevance, the function and the molecular mechanisms of the uPA system in PC metastasis. A current consensus suggests that the uPA system promotes PC metastasis by mediating pericellular plasminogen activation. Towards the development of therapeutic strategies that specifically target uPA-mediated PC metastasis, several remaining issues are discussed. PMID- 12085968 TI - Angiogenesis in prostate cancer: biology and therapeutic opportunities. AB - Tumor growth is limited in size without the incorporation of new blood vessels. Tumor cells release soluble factors (angiogenic factors) that induce neovascularization and allow subsequent growth beyond 2-3 mm in diameter meeting the need for cellular uptake of oxygen and nutrients. This process is referred to as the 'angiogenic switch' and indicates the acquisition of an angiogenic phenotype. Tumor angiogenesis requires an imbalance between proangiogenic and antiangiogenic factors with formation of new vessels being a highly regulated process. In this review we discuss the mediators of angiogenesis, the strategies for manipulating angiogenic factors, and possible therapeutic applications with a special emphasis on prostate cancer. PMID- 12085969 TI - Integrins and prostate cancer metastases. AB - Integrins have emerged as modulators of a variety of cellular functions. They have been implicated in cell migration, survival, normal and aberrant cellular growth, differentiation, gene expression, and modulation of intracellular signal transduction pathways. In this review article, the structural and functional characteristics of integrins, their expression and their potential role in prostate cancer metastases will be discussed. PMID- 12085970 TI - Prostate carcinoma skeletal metastases: cross-talk between tumor and bone. AB - The majority of men with progressive prostate cancer develop metastases with the skeleton being the most prevalent metastatic site. Unlike many other tumors that metastasize to bone and form osteolytic lesions, prostate carcinomas form osteoblastic lesions. However, histological evaluation of these lesions reveals the presence of underlying osteoclastic activity. These lesions are painful, resulting in diminished quality of life of the patient. There is emerging evidence that prostate carcinomas establish and thrive in the skeleton due to cross-talk between the bone microenvironment and tumor cells. Bone provides chemotactic factors, adhesion factors, and growth factors that allow the prostate carcinoma cells to target and proliferate in the skeleton. The prostate carcinoma cells reciprocate through production of osteoblastic and osteolytic factors that modulate bone remodeling. The prostate carcinoma-induced osteolysis promotes release of the many growth factors within the bone extracellular matrix thus further enhancing the progression of the metastases. This review focuses on the interaction between the bone and the prostate carcinoma cells that allow for development and progression of prostate carcinoma skeletal metastases. PMID- 12085971 TI - Signal transduction targets in androgen-independent prostate cancer. AB - Prostate cancer (PCa) first manifests as an androgen-dependent disease. Thus, androgen-deprivation therapy is a standard regimen for patients with metastatic PCa. Despite the initial success of androgen-deprivation therapy, PCa inevitably progresses from being androgen dependent (AD) to androgen independent (AI), and this marks the poor prognosis of this disease. Relapse of AIPCa becomes life threatening and accounts for the majority of mortality of PCa patients. Currently, no effective therapy is available for controlling AIPCa. Therefore, the challenge in providing a new intervention is to understand the fundamental changes that occur in AIPCa. Increasing evidence indicates that, under androgen deprived milieu, several signal networks elicited by peptide growth factors dictate the AI phenotype of PCa. This review covers the latest studies investigating the potential involvement of autocrine growth factors in cell proliferation, survival, metastasis, and the reciprocal interaction with the androgen receptor pathway. In addition, loss of the negative feedback mechanism of the signal cascade further amplifies the effect of growth factors, and thus contributes significantly to the onset of AIPCa. The understanding of the signal target(s) in AIPCa should provide the new markers for prognosis and a new strategy for prevention and therapy. PMID- 12085972 TI - Sudden death in young athletes: HCM or ARVC? AB - Sudden non-traumatic death in young athletes is due to underlying congenital/inherited cardiac diseases in over 80% of cases. The two commonest conditions leading to sudden cardiac death in athletes below the age of 25 years are hypertrophic cardiomyopathy (HCM) and arrhythmogenic right ventricular cardiomyopathy (ARVC). Hypertrophic cardiomyopathy is caused by mutations in genes, which code for sarcomeric contractile proteins. It can present with symptoms such as palpitation, presyncope or syncope. In a small number of cases, sudden death is the first clinical manifestation of the condition. It is well established that HCM accounts for over half of all cases sudden cardiac death in young individuals below 25 years of age. The management of HCM broadly encompasses symptom control, familial evaluation and the prevention of sudden death. Arrhythmogenic right ventricular cardiomyopathy, similarly, is a genetic disorder of the heart muscle and leads to symptoms such as palpitation and syncope and more rarely sudden death. The diagnosis of ARVC is most likely underestimated due to the lack of a single diagnostic test and subtle morphological changes in some cases. The diagnosis is based on clinical and family history and non-invasive investigations. The physiological adaptations seen in some athletes, as a response to physical training, may resemble phenotypically mild forms HCM and ARVC. Therefore, a diagnostic algorithm enabling this differentiation would be of importance especially bearing in mind the consequences of a misdiagnosis. PMID- 12085973 TI - Romano-Ward and other congenital long QT syndromes. AB - Molecular genetics applied to the study of inherited arrhythmogenic diseases has profoundly modified our understanding of cardiac electrophysiology providing new information on the crucial pathophysiological role of cardiac ion channels. These data are now putting forth innovative strategies for clinical management of the affected patients. Among these conditions, long QT syndrome (LQTS) was the first to enter the "genetic era", and nowadays the availability of large population of patients with known mutation allows to draw meaningful genotype-phenotype correlation and genetic-based risk stratification. However, despite the remarkable impact on knowledge, several still poorly defined issues limit the translation of such information into more effective therapeutic stratigies. As an example, despite the evidence of a significant QT shortening potential, the gene specific therapy of LQTS has still to prove its impact upon the risk of cardiac events. The present article reviews the most critical findings obtained in the last decade in the field of genetic of LQTS in the attempt of underlying its current applicability, limitations and the future perspectives of this knowledge in the management of affected patients. PMID- 12085974 TI - The syndrome of right bundle branch block, ST segment elevation in V1 to V3 and sudden death. PMID- 12085975 TI - Effects of quinapril on myocardial function, ventricular remodeling and cardiac cytokine expression in congestive heart failure in the rat. AB - Inflammatory cytokines have been shown to be activated in congestive heart failure (CHF). This activation is likely the result of the convergence of a number of factors, several of which could be attenuated with the use of an Angiotensin converting enzyme (ACE) inhibitor. In order to assess this, rats had a myocardial infarction (MI) created by coronary artery ligation and were followed for 28 days without treatment to permit the development of CHF. At that time, the ACE inhibitor quinapril was started, or rats remained untreated and were followed a further 56 days for a total of 84 days. Half of the untreated rats had quinapril started 3 days prior to sacrifice, on day 81. Starting quinapril at either 28 or 81 days had little effect on cardiac hemodynamics, or ventricular remodeling. Quinapril did however attenuate the MI-induced rise in cardiac cytokine expression (tumor necrosis factor-alpha [TNF-alpha], interleukin 1beta, -5 and -6). Thus, in CHF, ACE inhibitors attenuate the rise in cardiac cytokine expression. This study helps to identify a new mechanism by which ACE inhibitors may exert their beneficial effects in CHF. PMID- 12085976 TI - Cardioprotective effect of pump prime aprotinin in coronary artery bypass grafting. AB - BACKGROUND: Several studies have reported that high-dose aprotinin is cardioprotective in coronary surgery. The cardioprotective efficacy of low-dose aprotinin is less well defined. The present randomised study evaluated the cardioprotective and anti-inflammatory effects of pump prime aprotinin in patients undergoing coronary bypass surgery. METHODS: Sixty-four male patients admitted for first-time elective coronary artery bypass surgery were randomised into control or aprotinin groups. Patients in the aprotinin group received 280 mg of aprotinin in the pump prime. Postoperative CK-MB release, leukocyte counts and hemodynamics were recorded. Perioperative myeloperoxidase, IL-6, IL-8 and IL-10 levels were measured in a subgroup of patients (15 patients in each group). RESULTS: There were no significant differences between the groups in mechanical ventilation time and ICU and hospital stay. Postoperative bleeding was less serious in the aprotinin group than in the controls (742.0 +/- 361.1 versus 885.2 +/- 335.1 ml, p = 0.12) and CK-MB values were significantly lower (6 hrs, 35.5 +/ 11.8 versus 4.5 +/- 24.0 U/L; 24 hrs, 32.3 +/- 25.0 versus 40.2 +/- 26.8 U/L; 48 hrs, 15.9 +/- 7.0 versus 24.7 +/- 21.1 U/L; p=0.041). Perioperative hemodynamics was similar in both groups. There was a tendency towards less vasopressor and inotropes use in the pump prime aprotinin group. There was no significant difference between groups in terms of perioperative myeloperoxidase, IL-6, IL-8 and IL-10 levels. CONCLUSIONS: Pump prime aprotinin marginally limits myocardial enzyme release, but fails to limit inflammatory responses after elective coronary surgery. PMID- 12085977 TI - Effect of nipradilol on silent myocardial ischemia and heart rate variability in chronic stable angina. AB - PURPOSE: Silent myocardial ischemic episodes as well as decreased heart rate variability (HRV) indices are associated with an unfavorable outcome in patients with coronary artery disease. Nipradilol, which is a nonselective beta-adrenergic and nitrate-like vasodilator anti-anginal agent developed in Japan, may ameliorate silent myocardial ischemia, while it also improves exercise tolerance and HRV indices in patients with chronic stable angina. METHODS: To investigate the effect of nipradilol (6 mg daily) on silent myocardial ischemic episodes and HRV indices, and to study its effect on the relationship between them, 24 patients with chronic stable angina underwent exercise treadmill testing and a 24 hour ambulatory electrocardiogram (ECG). The study protocol utilized a single blind, 4-week placebo-controlled design. The HRV indices from ambulatory ECG included mean RR (ms), SDNN (ms), SDANN (ms), SD (ms), rMSSD (ms), pNN50 (%); frequency analysis of HRV consisted of total (ms, 0.01-1.00 Hz), low (ms, 0.04 0.15 Hz) and high (ms, 0.15-0.40 Hz) components. RESULTS: Nipradilol significantly decreased the mean heart rate at submaximal and maximal exercise and the mean pressure rate product at submaximal and maximal exercise. It significantly improved exercise-induced maximal ST segment depression from -1.7 +/- 0.6 mm to -1.1 +/- 0.7 mm (p < 0.05). Silent myocardial ischemic episodes recorded during the 24-hour ambulatory ECG significantly decreased after nipradilol administration. Nipradilol also significantly influenced several HRV indices as well as the relationship between silent myocardial ischemic episodes and the HRV indices. Nipradilol significantly increased SD, rMSSD, pNN50, total spectra, low frequency spectra and high frequency spectra. In addition, nipradilol significantly decreased the LF/HF ratio from 1.7 (1.5-2.0) to 1.5 (1.3 1.8). These effects of nipradilol on HRV indices concomitantly occurred with the reduction in silent myocardial ischemic episodes. CONCLUSION: Nipradilol was found to effectively improve the episodes of silent myocardial ischemia as well as exercise-induced ischemia probably due to its beta-blocking properties and not nitrate-like actions. In addition, nipradilol also had a favorable effect on the HRV indices. PMID- 12085978 TI - Melatonin protects the heart against both ischemia/reperfusion injury and chemotherapeutic drugs. PMID- 12085979 TI - Economic evaluation of the randomized aldactone evaluation study (RALES): treatment of patients with severe heart failure. AB - PURPOSE: To use data from the Randomized Aldactone Evaluation Study (RALES) to compare clinical outcomes and costs as part of the assessment of the economic implications of spironolactone treatment of advanced heart failure. METHODS: RALES was a randomized, double-blinded, placebo-controlled trial that enrolled participants who had severe heart failure and a left ventricular ejection fraction of no more than 35% and who were receiving standard therapy, including an angiotensin-converting enzyme inhibitor, a loop diuretic, and, in some cases, digoxin. We used a decision analytic model that incorporated data from participants in RALES as well as cost data from five countries that participated in the study. Costs were calculated for nonfatal hospitalizations, ambulatory care, spironolactone therapy, and death. The primary health outcome was quality adjusted life-years saved (QALYS). Outcomes were evaluated for the first 35 months of observation in RALES. RESULTS: Spironolactone therapy during the first 35 months of follow-up in RALES increased quality-adjusted survival time (0.13 QALYS, 95% CI, 0.07 to 0.18) without increasing costs ($713 savings, 95% CI, $2,123 savings to $783 in costs). Spironolactone therapy either dominated placebo or had a ratio of cost per QALYS that was unlikely to exceed $20,300. These results were robust in both one-way and multiway sensitivity analyses. CONCLUSIONS: Even after implementation of current clinical guidelines, addition of spironolactone therapy provides an opportunity to further reduce the large clinical and economic burden of patients with heart failure. PMID- 12085980 TI - The economic efficiency of amlodipine in the treatment of coronary atherosclerosis--an analysis based on the PREVENT study. AB - BACKGROUND: High health service expenditure on the one hand, and the politically declared objective of stability of statutory contributions and restriction of public funds on the other hand, have been central points of the political and social discussion for several years. The over-proportional increase in health service expenditure is obvious, compared to the increase in the costs of living. PATIENTS AND METHODS: Cost-effectiveness of amlodipine in the treatment of coronary atherosclerosis was analysed by applying the results of the PREVENT study to the Swiss health system. RESULTS: Calculation of effectiveness shows an additional life expectancy of 0.083 years in the amlodipine cohort compared to the placebo cohort, over an observation period of 3 years. The cost-effectiveness of amlodipine treatment is approximately CHF 14,650 per life-year gained. CONCLUSION: The administration of the calcium antagonist amlodipine in CHD patients is cost-effective. PMID- 12085981 TI - Angiotensin converting enzyme inhibition in heart failure: clinical trials and clinical practice. AB - Heart failure is a common and often debilitating condition, but one for which there exists a variety of effective pharmacological therapies. The angiotensin converting enzyme (ACE) inhibitors represent one of the mainstays of the treatment of heart failure. In spite of a wealth of evidence regarding the efficacy of these agents in improving mortality and morbidity in heart failure, they are often under-utilised. Failure to prescribe or to prematurely withdraw ACE inhibitor therapy often stems from physicians perceptions regarding the likelihood of unwanted effects, in particular hypotension, renal impairment and cough. The evidence from clinical trials is that these unwanted effects are relatively uncommon. In routine clinical practice the rate of prescription of ACE inhibitor therapy is related to the expertise and motivation of the physician. There is a need for education of all health care professionals involved in the care of patients with heart failure with regard to the maximisation of ACE inhibitor therapy in heart failure. PMID- 12085982 TI - Angiotensin II as an inflammatory mediator: evolving concepts in the role of the renin angiotensin system in the failing heart. PMID- 12085983 TI - Clinical trials and the real world: selection bias and generalisability of trial results. AB - The clinical effectiveness movement underpins a major health professional culture change which is occurring worldwide. Clinical practice guidelines are an important component of this movement and seek to promote evidence-based care, and combine quality and efficiency with equity of access in a patient centred approach. However some areas of practice are more amenable to guidelines than others and may thus be advantaged in terms of resource allocation. While high grade evidence from randomised controlled trials forms the basis for many cardiovascular guidelines, selection bias may considerably limit the generalisability of such data. This is particularly exemplified by clinical trials in heart failure which are not necessarily relevant to the majority of older patients with heart failure in the community. Future research in heart failure should be oriented towards the community and realigned in the context of health care priorities for the more elderly who have yet to benefit from previous advances. PMID- 12085984 TI - Antioxidant activity of extracts from Euryale ferox seed. AB - Euryale ferox has been widely used in traditional oriental medicine to treat a variety of illness. However, very little is known about the cellular actions by which this plant mediates its therapeutic effects. Various aspects of antioxidant activity were evaluated in total extracts and fractions derived from Euryale ferox. Total extracts (IC50 5.6 microg/ml) showed relatively high level radical scavenging activity toward 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and also enhanced viability of Chinese hamster lung fibroblast (V79-4) cells under exposure to oxidative agents. Upon further fractionation, the highest levels of DPPH radical scavenging and lipid peroxidation inhibitory activities were found in the ethyl acetate and butanol fractions. The ethyl acetate fractions, the butanol fractions, and total extracts of Euryale ferox also dose-dependently enhanced the activities of superoxide dismutase, catalase and glutathione peroxidase in V79-4 cells. Of these three antioxidant enzymes, glutathione peroxidase activity was most strongly induced. Taken together, our findings show that Euryale ferox contains a significant antioxidant activity and that specific components in the ethyl acetate and butanol fractions may play an important role in mediating these antioxidant properties. PMID- 12085985 TI - The effect of telomerase expression on the escape from M2 crisis in virus transformed human retinal pigment epithelial cells. AB - Transformation with viral oncogene extends the lifespan of normal cells beyond replicative senescence called M1, but most of them eventually succumb to second crisis called M2 when telomeres become critically short. To acquire an infinite growth capacity, these cells have to overcome M2 crisis, which is known to follow telomerase activation. We have investigated if telomerase expression is required for virus-transformed pre-M2 cells to avert M2 crisis. Human retinal pigment epithelial (RPE) cells were transformed with simian virus 40 large T antigen and a VR3 clone in pre-M2 stage was obtained. Then, VR3 cells were transfected with a telomerase-containing vector and two cell lines that expressed telomerase temporarily or continuously were cloned and designated as ST1 and ST2, respectively. Normal RPE cells went into senescence after 36 population doublings. Although the lifespan was extended in the VR3 clone about 20 times more, it eventually underwent second crisis. The telomere length of VR3 decreased compared to that of normal RPE cells and the decrease continued during subculture. However, the ST1 and ST2 clones that expressed both T antigen and telomerase could avert this crisis. The initial telomere length of ST1 and ST2 was longer than that of normal cells. The ST1 underwent growth arrest again as telomerase expression faded out and elongated telomere was shortened, but the ST2 that maintained telomerase activity and telomere length proliferated continuously. In conclusion, telomerase activation is definitely required to overcome M2 crisis and acquire an infinite lifespan in human somatic epithelial cells and this mechanism is independent from M1 crisis escape in cell immortalization. PMID- 12085986 TI - Human hepatocellular carcinoma cells resist to TRAIL-induced apoptosis, and the resistance is abolished by cisplatin. AB - TNF-related apoptosis-inducing ligand (TRAIL), a member of the TNF family, selectively induce apoptosis in various transformed cell lines but not in almost normal tissues. It is regulated by 2 death receptors, TRAIL receptor 1 (TRAIL-R1) and TRAIL-R2 and 2 decoy receptors, TRAIL-R3 and TRAIL-R4. However, the determining factors of the sensitivity to TRAIL-induced apoptosis are not clearly understood. Herein, we investigated the expression of TRAIL-R, c-FLIP, FADD-like interleukin-1beta-converting enzyme inhibitory protein, and TRAIL-induced apoptosis in human hepatocellular carcinoma (HCC) cell lines. Seven of ten HCC cell lines showed resistance to TRAIL-induced apoptosis and five of seven TRAIL resistant cell lines became sensitive to TRAIL by co-treatment with cycloheximide. In HCC cell lines, their TRAIL resistance did not correlate with the basal expression level of TRAIL receptors or c-FLIP, however, in human tissues, TRAIL-R1 and TRAIL-R2 expressions were notably decreased compared to normal counterpart. Cisplatin showed synergistic effect on TRAIL-induced apoptosis in most HCC cell lines regardless of their p53 status and TRAIL-R1 was induced by cisplatin treatment in certain cell lines. Inhibition of nuclear factor K B (NF-kappaB) by SN50, a peptide inhibitor of NF-KB activity, had no effect on TRAIL-induced apoptosis in HCC cells. These results suggest that (a) the majority of human HCC cell lines are resistant to TRAIL-induced apoptosis and cycloheximide-sensitive short-lived antiapoptotic molecule(s) is responsible for this resistance, (b) the expression of TRAIL-R1 and TRAIL-R2 is reduced in HCC tissues, and the increased expression of TRAIL-R1 may be a mechanism of cisplatininduced sensitization to TRAIL-induced apoptosis in some HCC cells, and (c) the activation of NF-kappaB may not be involved in the TRAIL resistance of HCC cells PMID- 12085987 TI - Effects of high taurocholate load on activities of hepatic alcohol metabolizing enzymes. AB - Membrane-associated cytotoxicity induced by hydrophobic bile salts is a major contributing factor leading to liver diseases. Administration of ursodeoxycholate reduces serum liver enzymes in chronic liver diseases but the nature of this effect is still unclear. Using alcohol metabolising enzymes as cellular markers, the hepatotoxic properties of hydrophobic bile salts and the putative hepatoprotective effect of ursodeoxycholate was examined. Two animal models of biliary retention, bile duct obstruction and choledochocaval fistula was used to investigate the effect of taurocholate on the hepatic alcohol metabolizing enzymes: cytosolic alcohol dehydrogenase, microsomal ethanol oxidizing system, catalase and aldehyde dehydrogenase before and after the infusion of taurocholic acid or tauroursodeoxycholic acid for two days period. Bile duct obstruction was found to be similar to or slightly exceeds choledochocaval fistula in the degree of retention. Following the taurocholic acid infusion, the serum alcohol dehydrogenase activity as well as microsomal ethanol oxidizing system and aldehyde dehydrogenase were greatly increased but the level of cytosolic alcohol dehydrogenase and catalase activities was found to be lower in either or both models in comparison with the control animals. However, the tauroursodeoxycholic acid infusion did not induce any significant changes in the levels of all the alcohol metabolizing enzyme activities in either or both models. These findings suggest that hydrophobic taurocholic acid (7alpha) affects the plasmalemma to allow leakage of cytosolic alcohol dehydrogenase into the blood circulation, stimulates the biosynthesis of microsomal ethanol oxidizing system and aldehyde dehydrogenase, and suppresses the biosynthesis of alcohol dehydrogenase and catalase. But in contrast, the hydrophilic tauroursodeoxycholic acid (7beta) provided hepatoprotective effect. PMID- 12085988 TI - Differential expression of two stress-inducible hsp70 genes by various stressors. AB - Inducible HSP70s encoded by two almost identical hsp70 genes, known as hsp70.1 and hsp70.3 are located in a tandem array on mouse chromosome 17. Duplication of this gene has been found in various higher organisms. However, the role of two almost identical hsp70 genes is still unclear. To elucidate the ambiguity of expression pattern between hsp70.1 and hsp70.3, we investigated the induction of each hsp70 isoforms by several HSP70-inducible stressors in wild type (hsp70.1+/+ and hsp70.3+/+) and the hsp70.1-/- (hsp70.1-/- and hsp70.3+/+) murine embryo fibroblast (MEF) cells, and the M-1 mouse cortical collecting duct cell line. Each induced hsp70 isoforms by heat shock were very similar at transcriptional and translational levels in wild type and hsp70.1-/- MEF cells. The mRNA stabilities of both hsp70.1 and hsp70.3, even in two kinds of hsp70.3 transcripts, were also very similar. L-azetidine-2-carboxylic acid, sodium arsenite, CdCl2 and ZnCI2 caused induction of both isoforms of the hsp70 genes, even though their expression levels were variable. NaCl caused induction of just hsp70.1 gene expression. H2O2 failed to induce the expression of either hsp70 genes in MEF cells, caused induction of hsp70.1 gene expression in the M-1 cell line. Hsp70 accumulation by H2O2 and NaCl treatment was mainly due to hsp70.1 expression. Our studies demonstrated that two hsp70 genes respond differentially to types of stress. PMID- 12085989 TI - Induction of hepatic inducible nitric oxide synthase by cholesterol in vivo and in vitro. AB - Cholesterol-rich diet impairs endothelial NO synthase (eNOS) and enhances inducible NOS (iNOS) expression. In this study, we investigated effects of cholesterol on iNOS expression in high-fat-fed rat models, HepG2 and RAW264.7 cells. The high-fat diet increased the plasma total cholesterol level 6-7 fold and low-density lipoprotein cholesterol level (LDL-C) approximately 70 fold and slightly increased the level of lipid peroxidation as determined by thiobarbituric acid-reactive substance assay. The high-fat diet also increased plasma nitric oxide (NO) concentrations up to 5 fold, and induced iNOS mRNA expression in liver. The contractile responses of the endothelium-denuded thoracic aortic rings to phenylephrine were significantly damaged in high-fat-fed rats when assessed by organ chamber study. Treatment with estrogen for 4 days failed to reduce iNOS expressions as well as aortic contractility, although it improved lipid profiles. In cultured HepG2 or murine macrophage RAW264.7 cells, 3 days treatment with either 25-hydroxycholesterol or 7-ketocholesterol induced iNOS mRNA expression, as determined by RT-PCR. Our data suggested that the chronic exposure of hepatocytes and macrophage cells to high concentration of cholesterol or oxysterols may induce iNOS expression and subsequent synthesis of NO, which may be important in the pathogenesis of atherosclerosis. PMID- 12085990 TI - Inhibitory action of bisphosphonates on bone resorption does not involve the regulation of RANKL and OPG expression. AB - The mechanism of inhibitory action of bisphosphonates on bone resorption is not fully elucidated. Osteoclast formation and activity are regulated by osteoblast derived factors such as the osteoclast differentiating factor, receptor activator of NF-kappaB ligand (RANKL) and the inhibitor, osteoprotegerin (OPG). To investigate in vitro effects of bisphosphonates on mouse osteoblastic cells, we examined the expression levels of RANKL and OPG in the cells treated with alendronate or pamidronate (10(-8) approximately 10(-5) M) alone, or combined with 10 nM of 1,25-(OH)2VitD3 for 24 or 48 h. Various concentrations of alendronate and pamidronate did not change the mRNA expression of RANKL and OPG consistently irrespective of 1,25-(OH)2VitD3 presence. When added into cocultures of mouse osteoblastic cells and bone marrow cells, both alendronate and pamidronate inhibited osteoclast formation and bone resorption but failed to alter the RANKL and OPG mRNA expression. These results indicate that the inhibition of bone resorption by bisphosphonates is not mediated by the regulation of RANKL and OPG expression. PMID- 12085991 TI - Quantitative measurement of serum allergen-specific IgE on protein chip. AB - Type I allergy is an immunoglobulin E (IgE)-mediated hypersensitivity disease inflicting more than quarter of the world population. In order to identify allergen sources, skin provocation test and IgE serology was performed using allergen extracts. Such process identifies allergen-containing sources but cannot identify the disease-eliciting allergenic molecules. Recently, microarray technology has been developed for allergen-specific IgE detection using rolling circle amplification. This study was carried out to evaluate protein chip technology for the quantitative measurement and limits of sensitivity of multiple allergen-specific IgE by an immunofluorescence assay. Significance of positive calibrators was tested using purified human IgE. Dermatophagoides pteronyssinus (Dp), egg white, milk, soybean, and wheat were used as allergens and human serum albumin as negative control. Sensitivity and clinical efficacy of protein chip were evaluated using allergy immune serum for Dp. The fluorescent intensities for purified human IgE as calibrator were well correlated with the concentrations of human IgE. Two-fold dilution of serum allowed an optimal reaction with Dp (1 mg/ml) at which serum Dp-specific IgE levels by protein chip were compatible with those by UniCap. The sensitivity of protein chip in this study was found at level of 1 IU/ml of IgE. Dp-specific IgE levels by protein chip correlated well with those of UniCap by comparing 10 atopic dermatitis. Additional 18 sera were tested for above multiple antigens other than Dp and significant results were obtained for many antigens as well as Dp. These results indicated that spotting of heterogeneous protein mixture on protein chip and the quantitative measurement of serum allergen-specific IgE levels using immunofluorescence assay can be successfully applied in the clinical laboratory for the diagnosis of allergy and could be applied to diagnosis of autoimmune and infectious diseases PMID- 12085992 TI - Induction of Bis, a Bcl-2-binding protein, in reactive astrocytes of the rat hippocampus following kainic acid-induced seizure. AB - The expression of Bis (also called Bag-3), a Bcl-2-binding protein, was investigated in the rat kainic acid (KA) model of temporal lobe epilepsy. Western blot analysis showed a significant increase in the expression levels of Bis protein in the hippocampus following the systemic administration of KA. Bis immunoreactivity increased preferentially in the CA1 and CA3 regions, as well as in the hilar region of the dentate gyrus. Experiments with double immunofluorescence revealed that, in KA-administered rats, the cells expressing Bis were GFAP-expressing reactive astrocytes. The increase in Bis immunoreactivity was accompanied by increased Bcl-2 in reactive astrocytes in the striatum radiatum, whereas Bcl-2 immunoreactivity in pyramidal neurons was not affected. These results of the co-expression of Bis and Bcl-2 in reactive astrocytes in this seizure model suggest that Bis might modulate the glial reaction under excitotoxic brain injury, probably by interacting with Bcl-2. PMID- 12085993 TI - Basic fibroblast growth factor-induced translocation of p21-activated kinase to the membrane is independent of phospholipase C-gamma1 in the differentiation of PC12 cells. AB - p21-activated kinase (PAK) targeting to the plasma membrane is essential for PC12 cell neurite outgrowth. Phospholipase C-gamma1 (PLC-gamma1) can mediate the PAK translocation in response to growth factors, since PLC-gamma1 binds to both tyrosine-phosphorylated receptor tyrosine kinases and PAK through its SH2 and SH3 domain, respectively. In the present study, we examined a potential role for PLC gamma1 in the basic fibroblast growth factor (bFGF)-induced PAK translocation using stable PC12 cell lines that overexpress in a tetracycline-inducible manner either the wild-type FGFR-1 or the Y766F FGFR-1 mutant. Phosphatidylinositol hydrolysis was increased 6.5-fold in response to bFGF in the wild type cells but negligible in the mutant cells. The recombinant GST-PLC-gamma1 SH3 was able to bind to PAK1 but not GST alone. However, examination of PLC-gamma1 as an adaptor for translocation of PAK1 in cells showed that both cells transfected with pEGFP PAK1 was able to differentiate for 24 h, as visualized by laser confocal microscopy. Translocation of PAK1 to growth cones occurs at similar levels in both wild and mutant cells. These results suggest that a protein(s) other than PLC-gamma1 is functionally relevant for PAK targeting. PMID- 12085994 TI - Effects of estrogen receptor and estrogen on the chromatin structure in estrogen receptor stable transfectants. AB - The estrogen receptor (ER), a member of the nuclear hormone receptor superfamily recruits coactivators that modify local chromatin structure. Here we investigated the effect of the estrogen receptor and estrogen on the global chromatin structure and the local chromatin structure of the progesterone receptor gene during the process of transcriptional activation using Rat1+ER cells stably expressing the estrogen receptor. The total chromatin was more accessible to DNasel in Rat1+ER cells than in the parental estrogen receptor-negative Rat1 cells. After 18 h of estrogen treatment, total chromatin was more dispersed in Rat1+ER cells than in Rat1 cells. The chromatin structure of the progesterone receptor gene was more sensitive to DNasel in Rat1+ER cells than in Rat1 cells. However, the chromatin structure of the progesterone receptor gene did not change further on estrogen treatment. Our results suggest that under certain circumstances unoccupied estrogen receptors may play some role in reorganizing the repressive chromatin structure to induce gene activation. PMID- 12085995 TI - Coronary artery dynamics in vivo. AB - There is considerable evidence that the localization and evolution of vascular disease are mediated, at least in part, by mechanical factors. The mechanical environment of the coronary arteries, which are tethered to the beating heart, is influenced by cardiac motion; the motion of the vessels must be described quantitatively to characterize fully the mechanical forces acting on and in the vessel wall. Several techniques that have been used to characterize coronary artery dynamics from biplane cineangiograms are described and illustrated. There is considerable variability in dynamic geometric parameters from site to site along a vessel, between the right and left anterior descending arteries, and among individuals, consistent with the hypotheses that variations in stresses mediated by geometry and dynamics affect the localization of atherosclerosis and individual risk of coronary heart disease. The few frankly atherosclerotic vessels that have been examined exhibit high torsions in the neighborhood of lesions, an observation which may have etiologic or diagnostic implications. PMID- 12085996 TI - Effect of fluid shear stress on the permeability of the arterial endothelium. AB - The localization of atherosclerotic lesions is due, in part, to regional variations in the permeability of arterial endothelium to macromolecules. In turn, endothelial permeability may be influenced by fluid shear stresses. The spatial variation in endothelial permeability is reviewed and evidence for shear stress dependence upon permeability is presented. These results are examined in light of various signaling mechanisms that increase permeability by increasing the transport of water and macromolecules through the junctions separating endothelial cells. Signaling pathways cause a change in the dense peripheral band of actin and actin stress fibers or alter the phosphorylation of junction proteins which affects their ability to localize in junctions. Future directions to clarify the effect of shear stress on permeability are considered. PMID- 12085997 TI - Fluid dynamics, wall mechanics, and oxygen transfer in peripheral bypass anastomoses. AB - Intimal hyperplasia at vascular anastomoses seems to be promoted by altered flow conditions and stress distributions within the anastomotic region. In order to gain deeper insight into postoperative disease processes, and subsequently, to contribute to the development of improved vascular reconstructions, detailed studies, also on local flow dynamics and related mass transport and wall mechanical effects, are required. In context with in vivo studies, computer simulation based on casts of femoro-popliteal bypasses implanted into sheep were performed to analyze the flow dynamics, the oxygen transport, and the wall and suture mechanics in anatomically correct bypass configurations related to three established surgical techniques and resulting geometries (conventional type anastomosis, Taylor-patch and Miller-cuff anastomoses with venous interposition grafts of different modifications). The influence of geometry, compliance of the graft, the interponated vein patch and vein cuff, and of the artery was included. Time-dependent, three-dimensional Navier-Stokes equations describing the flow field, and a nonlinear shell structure for the vessel walls were coupled using finite element methods. The numerical results demonstrate nonphysiological flow patterns in the anastomotic region. Strongly skewed axial velocity profiles and secondary velocities occur in the junction region. In the Miller-cuff a vortex may induce a wash-out effect which protects the host artery. On the artery floor opposite the junction flow separation and zones of recirculation were found. The analysis of oxygen transport illustrates a correlation between zones of low wall shear stress and reduced oxygen flux into the wall. Wall mechanics show that increased compliance mismatch leads to increased and discontinuous intramural stresses. Comparison to histomorphological findings on intimal hyperplasia shows certain correlations, particularly increased compliance mismatch has a proliferate influence on suture line hyperplasia. The reduction of compliance mismatch using vein interposition results in decreased generation of intimal hyperplasia, and therefore, contributes to improvement of patency rates, while the geometrical modification and the resulting change of the flow pattern seems to be less important for the growth of anastomotic intimal hyperplasia. PMID- 12085999 TI - Role of subcellular shear-stress distributions in endothelial cell mechanotransduction. AB - The endothelium of blood vessels presents a wavy surface to the flowing blood. The subcellular distribution of shear stress depends on the shape and orientation of the cells and on their spatial arrangement within the monolayer. By studying details of the distribution of stress at this scale and the morphological responses that serve to modify the distribution, we can gain insight into the physical mechanisms by which the cell senses its fluid mechanical environment. The rapidly growing body of evidence indicates that endothelial cells discriminate between subtle variations in the exact loading conditions including differences in temporal and spatial gradients of shear stress, steady and pulsatile laminar flow, and laminar and turbulent flows. While in a few studies the effects of these individual flow characteristics have been carefully isolated, it is difficult to assess the relative importance of any one parameter. To interpret the relationships between isolated flow characteristics or the integrated effects of combined loading conditions and the biochemical signaling events that mediate the cell response, a full stress analysis of the cell is needed. The microscopic distribution of shear stress acting upon the cell surface provides the boundary condition for such an analysis. Experimental and analytical tools are being developed to assess the stress distribution throughout the cellular structures that might be involved in mechanotransduction. PMID- 12085998 TI - Computational modeling of mass transfer and links to atherosclerosis. AB - In the context of atherogenesis, mass transport refers to the movement of atherogenic molecules from flowing blood into the artery wall, or vice versa. Although LDL transport clearly plays a role in atherosclerotic plaque development, it is much less clear whether abnormalities in mass transfer patterns are in themselves atherogenic. A powerful way of addressing this question is through computational modeling, which provides detailed descriptions of local mass transport features. Here we briefly review the strategy and some of the pros and cons of such a modeling approach, and then focus on results gained from studies in a variety of arterial geometries. The general picture is that zones of hypoxia (low oxygen transport from blood to wall) and elevated LDL tend to colocalize with each other, and with areas of atherosclerotic lesion development and/or intimal thickening. The picture is complicated by the fact that such zones also tend to have "abnormal" wall shear stress patterns, which are also believed to be atherogenic. Taken together, these results suggest, but do not prove, a role for mass transport in atherogenesis. PMID- 12086000 TI - Image-based computational fluid dynamics modeling in realistic arterial geometries. AB - Local hemodynamics are an important factor in atherosclerosis, from the development of early lesions, to the assessment of stroke risk, to determining the ultimate fate of a mature plaque. Until recently, our understanding of arterial fluid dynamics and their relationship to atherosclerosis was limited by the use of idealized or averaged artery models. Recent advances in medical imaging, computerized image processing, and computational fluid dynamics (CFD) now make it possible to computationally reconstruct the time-varying, three dimensional blood flow patterns in anatomically realistic models. In this paper we review progress, made largely within the last five years, towards the routine use of anatomically realistic CFD models, derived from in vivo medical imaging, to elucidate the role of local hemodynamics in the development and progression of atherosclerosis in large arteries. In addition to describing various image-based CFD studies carried out to date, we review the medical imaging and image processing techniques available to acquire the necessary geometric and functional boundary conditions. Issues related to accuracy, precision, and modeling assumptions are also discussed. PMID- 12086001 TI - Fluid and solid mechanical implications of vascular stenting. AB - Vascular stents have emerged as an effective treatment for occlusive vascular disease. Despite their success and widespread use, outcomes for patients receiving stents are still hampered by thrombosis and restensosis. As arteries attempt to adapt to the mechanical changes created by stents, they may in fact create a new flow-limiting situation similar to that which they were intended to correct. In vitro fluid mechanics and solid mechanics studies of stented vessels have revealed important information about how stents alter the mechanical environment in the arteries into which they are placed. Adverse nonlaminar flow patterns have been demonstrated as well as remarkably high stress concentrations in the vessel wall. In vivo studies of stented vessels have also shown a strong relationship between stent design and their dynamic performance within arteries. Alterations in pressure and flow pulses distal to the stent have been observed, as well as regional changes in vascular compliance. Considering the influence of flow and stress on the vascular response and the suboptimal clinical outcomes associated with stenting, knowledge gained from stent/artery mechanics studies should play an increasingly important role in improving the long-term patency of these devices. PMID- 12086002 TI - Elastodynamics and arterial wall stress. AB - Recent advances in molecular and cell biology have emphasized the fundamental importance of mechanical factors in regulating the structure and function of cells and extracellular matrix in the vasculature. Consequently, there is an ever greater motivation to calculate accurately the stress and strain fields in the arterial wall and how they change with disease, injury, and clinical treatment. Although there is an extensive literature on arterial mechanics, our understanding is still far from complete. In this paper, we review some of the salient findings with regard to wall properties, suggest some possible improvements in the calculation of wall stress, and identify some unresolved problems for further research. PMID- 12086003 TI - Biomechanics of plaque rupture: progress, problems, and new frontiers. AB - Plaque rupture has become identified as a critical step in the evolution of arterial plaques, especially as clinically significant events occur in critical arteries. It has become common in the past dozen years or so to consider which plaques are vulnerable, even though not yet ruptured. Thrombotic events have remained significant, but in a context where they are seen as being triggered often by plaque rupture. Weaving together considerations from structural mechanics, fluid mechanics, plaque morphology, epidemiological pathology, micromechanical measurements of arterial wall tissues, and emerging information on the complex roles of the matrix metalloproteinases, this critical review draws attention to the relative paucity of data (i) on the mechanical behavior of small test portions of arterial tissues and (ii) on the relation of plaque locations to local vessel curvature and curvature flexure. This is especially important in the epicardial arteries, where combination of biplane angiograms and intravascular ultrasound (both becoming increasingly available in digital recordings) offer opportunities for clinical investigation, allied to biomechanics, to an extent previously not possible. Improved imaging and local tissue property assessments provide related opportunities for the carotid bifurcation. The discussion includes a proposal for developing an assessment scale for plaque vulnerability. PMID- 12086004 TI - A morphometric model of lung mechanics for time-domain analysis of alveolar pressures during mechanical ventilation. AB - In this study we propose, and implement in the time domain, an anatomically consistent model of the respiratory system in critical care conditions that allows us to evaluate the impact of different ventilator strategies as well as of constrictive pathologies on the time course of acinar pressures and flows. We discuss the simplifications of the original Horsfield structure (Horsfield, K., et al. Models of the human bronchial tree. J. Appl. Physiol. 31:207-217, 1971), which were needed to enable the model implementation. The model has a binary tree structure including large airways represented as a combination of wall compliance and laminar resistance, small airways that have the same arrangement but can be heterogeneously constricted, and alveolar compartments that are viscoelastic second-order models to represent the stress adaptation behavior of lung tissue. We have described patient-ventilator interactions modeling the ventilator and the endotracheal tube. In conclusion this model makes it possible to investigate realistically the effect of homogeneous versus heterogeneous constrictive pathologies and the impact of different ventilatory patterns on pressure and flow distribution at the acinar level in the mechanically ventilated patient. PMID- 12086005 TI - Finite element analysis of imposing femtonewton forces with micropipette aspiration. AB - A novel technique of imposing femtonewton forces with micropipette aspiration [i.e., the extended micropipette aspiration technique (EMAT)] is proposed, and an axisymmetric finite element analysis of this technique is provided. The EMAT is experimentally based upon a micropipette manipulation system and is theoretically based upon hydrodynamics. Any spherical object such as a human neutrophil or a latex bead can be employed as the force transducer, so cell-cell interactions can be directly studied. Our computational analysis shows that femtonewton forces can indeed be imposed. The force magnitude is sensitive to the radius of the micropipette and the micropipette-transducer distance, but it is much less sensitive to other parameters including the radius of the transducer, the substrate curvature, and the thickness of the micropipette wall. Combining the EMAT and the previously developed micropipette aspiration technique will allow us to impose an unprecedented range of forces, from a few femtonewtons to a few hundred piconewtons on single molecules or receptor-ligand bonds. PMID- 12086006 TI - Modeling of nonlinear physiological systems with fast and slow dynamics. II. Application to cerebral autoregulation. AB - Dynamic autoregulation of cerebral hemodynamics in healthy humans is studied using the novel methodology of the Laguerre-Volterra network for systems with fast and slow dynamics (Mitsis, G. D., and V. Z. Marmarelis, Ann. Biomed. Eng. 30:272-281, 2002). Since cerebral autoregulation is mediated by various physiological mechanisms with significantly different time constants, it is used to demonstrate the efficacy of the new method. Results are presented in the time and frequency domains and reveal that cerebral autoregulation is a nonlinear and dynamic (frequency-dependent) system with considerable nonstationarities. Quantification of the latter reveals greater variability in specific frequency bands for each subject in the low and middle frequency range (below 0.1 Hz). The nonlinear dynamics are prominent also in the low and middle frequency ranges, where the frequency response of the system exhibits reduced gain. PMID- 12086007 TI - Three-layer volume conductor model and software package for applications in surface electromyography. AB - On comparing multichannel surface electromyographic measurements of the m. biceps brachii with simulations performed with a previously developed two-layer volume conduction model, we found substantial discrepancies. To incorporate an apparent distorting effect of the skin tissue, the model was extended to three layers. This new model describes the potential resulting from an eccentric bioelectric source in a finite, cylindrical, and anisotropic volume conductor consisting of three layers, representing muscle, subcutaneous fat, and skin tissue. This contribution presents the governing mathematical equations of the three-layer volume conductor model as well as our approach to their solution. A comparison of various models shows that the three-layer model best describes measured potential distributions. Furthermore, we present the "ANVOLCON" (analytical volume conductor) software package. This package has been developed to facilitate the use of the model for scientific and educational purposes and is freely available from http://www.mbfys.kun.nl/knf PMID- 12086008 TI - Comparison of algorithms for the simulation of action potentials with stochastic sodium channels. AB - This paper presents a comparison of computational algorithms to simulate action potentials using stochastic sodium channels. Four algorithms are compared in single-node models: Strassberg and DeFelice (1993) (SD), Rubinstein (1995) (R), Chow and White (1996) (CW), and Fox (1997) (F). Neural responses are simulated to a simple and a preconditioned monophasic current pulse. Three exact algorithms implementing Markov jumping processes (SD, R, CW) resulted in similar responses, while the approximation algorithm using Langevin's equation (F) showed quite different responses from those in the exact algorithms. The computational time was measured as well: 1(F), 7(CW), 32(SD), 39(R) relative to that of the F algorithm. Furthermore, it is shown that as the sampling step for integration of the transmembrane potential increases, neural responses in all algorithms tended to be different from those in dense sampling steps, however, the CW algorithm was robust even at a sparse sampling step. It is concluded that the most computationally efficient algorithm having appropriate properties of neural excitability is the CW algorithm. PMID- 12086010 TI - Community integration and statewide systems change: qualitative evaluation research in community life and disability. AB - As the disability field moves to the next generation of community life, this article describes the findings of a qualitative state policy research study on deinstitutionalization and community integration. New Hampshire was selected as a leading national example in the United States based, in part, upon the closure of its only public institution showing community leadership in the field of mental retardation and developmental disabilities. Drawing on the disciplines of politicial science, sociology, psychology and rehabilitation (Majchrzak, 1980), the research design uses a multi-case, multi-site study approach (Yin, 1989), and on-site, semi-structured interviewing of key informants using a research field guide (Taylor & Bogdan, 1984/1998). The qualitative research reflects an indepth version of state formative evaluation research studies on community integration (e.g, Taylor, Racino, & Rothenberg, 1988), and the use of coded data and multiple analytic techniques (e.g., Glaser & Strauss, 1967) congruent with the emerging support and empowerment paradigm (Racino, 1992). The article highlights four sets of research findings (i.e., state characteristics and community integration practices, thematic case studies of change, comparative roles in the change process, and a theoretical framework for understanding change) which form the basis for further statewide study of change toward community life in the United States. PMID- 12086011 TI - Economic, political and ethnic influences on end-of-life decision-making: a decade in review. AB - Implementation of the Patient Self Determination Act (PSDA) a decade ago has been followed by considerable research examining usage of advance directives among patient populations as well as among health care professionals. This article summarizes the history of end-of-life medical treatment options and provides a discussion of critical economic, political, and legal events that preceded passage of the PSDA. A review of the literature during the decade that followed is presented including research exploring ethnic factors in the completion of advance directives. Barriers to completing advance directives by minority populations are discussed and considerations for policy makers are provided. PMID- 12086009 TI - Wavelet-based multifractal analysis of human balance. AB - Center of pressure (COP) traces have been used to investigate the dynamics of human balance. In this paper we employ a wavelet-based multifractal methodology to identify structural differences in mediolateral and anterioposterior sway between COP traces of healthy and Parkinson's patients. Two statistical techniques are used to summarize the differences in multifractal spectrum (MFS) for both groups. The first technique is a multivariate repeated measures analysis on estimated MF spectra for subjects. The second technique obtains two characteristic measures from each subject's estimated MFS: (i) location and (ii) half-width of the spectrum. These measures present an intuitive summary of the MFS for each subject, allowing for statistical comparisons between the two groups. Both analyzes lead to significant discrimination between Parkinson versus healthy subject's MFS. We find that COP time series of Parkinson patients exhibit a greater degree of roughness as compared to healthy subjects' COP traces. Furthermore, MFS for Parkinson patients are narrower, suggesting a reduction in complexity as compared to the healthy group. The methodology presented here may be helpful in development of clinically relevant measures, including the assessment of severity of conditions as the measures developed here correlate with standard severity measures. PMID- 12086012 TI - Low-care residents in nursing homes: the impact of market characteristics. AB - Approximately 30% of nursing home residents were recently identified as low-care cases; that is, residents with low levels of acuity. Other institutional venues, board and care homes and assisted living facilities, for example, are often recommended as alternative domiciliaries providing more appropriate and less expensive care for these residents. In this investigation the effect of nine market factors on the prevalence of low-care residents in 14,646 nursing homes are studied. Government regulations, competition from other providers, and the overall munificence of the market are found to influence their prevalence. These results are discussed along with several issues inherent to channeling low-care residents to other care setting. PMID- 12086013 TI - Mandatory HIV testing of newborns in New York State: what are the implications? AB - This paper presents an analysis of a public health law (Maternal-Pediatric HIV Prevention and Care Program) enacted by New York State in 1997 and how it strives to reduce the vertical transmission of AIDS. This policy is twofold. First, it strongly encourages voluntary HIV testing of all pregnant women. Secondly, it mandates HIV testing for all newborns. Those found to be positive are referred for medical care. This paper analyzes the State's implementation of this law to determine if it will: exert control over poor, mostly minority women; provide health care for infected woman and infants; protect public health; prevent the spread of HIV/AIDS; or sacrifice a woman's privacy. The potential benefits and shortfalls of this policy are discussed as other states may consider adopting similar legislation. PMID- 12086014 TI - Enhancement by cyclosporin A of taxol-induced apoptosis of human urinary bladder cancer cells. AB - Taxol is a microtubule-stabilizing agent which induces apoptosis in various cancer cells. In this study, we found that T24 cells derived from high grade human urinary bladder cancer were relatively resistant to taxol and that the IC50 value determined by a colorimetric WST-1 assay was 406.0 nM. Interestingly, cyclosporin A (CsA), an immunosuppressive drug, dramatically enhanced sensitivity to taxol, and the IC50 value was decreased to 47.5 nM in the presence of 1 microM CsA. KK47 cells derived from low grade human urinary bladder cancer showed high sensitivity to taxol with an IC50 value of 78.8 nM which decreased to 14.4 nM in the presence of 1 microM CsA. FK506, another immunosuppressive drug, also enhanced sensitivity to taxol. Furthermore, a concomitant loss of calcineurin activity was observed after the treatment of both cell lines with both CsA and FK506. Taxol induced apoptosis of the cells, as assessed by Hoechst 33258 staining and by the measurement of caspase 3 activity. Immunoblot analysis with an antibody against Bcl-2 phosphorylated at serine 70 demonstrated that taxol induced the phosphorylation of Bcl-2 with its enhancement in the presence of CsA. In addition, treatment of the cells with CsA significantly decreased the expression of Bcl-2 at both the protein and mRNA levels. These results suggest that the enhancement of taxol-induced apoptosis by immunosuppressive drugs is at least partly due to the inhibition of calcineurin activity and the loss of the antiapoptotic function of Bcl-2 via the enhancement of phosphorylation and the reduction of expression. PMID- 12086015 TI - Thymidine phosphorylase activity in renal cell carcinoma: relationship between histological parameters and chemosensitivity to fluorouracil-related drugs. AB - Thymidine phosphorylase (TdR-Pase) is an essential enzyme in the metabolism of fluorouracil-related drugs and is also a potent angiogenic factor. We measured the TdR-Pase activity in human renal cell carcinomas (RCCs) to assess the relationship between TdR-Pase activity and the degree of tumor malignancy. We also investigated the relationship between TdR-Pase activity and chemosensitivity to fluorouracil-related drugs. A total of 61 RCC tissue specimens and 39 normal kidney tissue specimens were obtained. TdR-Pase activity was measured by enzyme linked immunosorbent assay. The in vitro histoculture drug response assay was also performed to examine tumor sensitivity to 5-fluorouracil (5-FU) and doxifluridine (5'-DFUR). The TdR-Pase activity of RCCs was 13-fold that of the normal tissues. Activity increased with histological grade, and,was significantly higher in high-stage tumors (T3 or higher). TdR-Pase activity exhibited a significant positive correlation with sensitivity to 5-FU and 5'-DFUR. High TdR Pase activity in human RCC was confirmed to predict high tumor grade and stage. RCCs with high TdR-Pase activity were also sensitive to FU-related drugs. PMID- 12086016 TI - A possible regulatory role of glyoxalase I in cell viability of human prostate cancer. AB - A role of glyoxalase I (Gly-I), a detoxifying enzyme, in cell viability of prostate cancer was investigated. Cell extracts obtained from 66 prostate tissue specimens and prostatic cancer PC-3 cells were assayed for Gly-I activity using the spectrophotometric method. Gly-I activity was consistently more than eightfold higher in prostate cancer (CAP) specimens (n = 37) than in non cancerous (NCP) specimens (n = 29). To understand the importance of such a high Gly-I activity in CAP specimens, the effects of methylglyoxal (MG) on PC-3 cells were examined in vitro. MG, a putative toxic glycolytic metabolite, was capable of inducing severe (> 99%) cell death in 24 h, along with a significant reduction in activities of Gly-I as well as glyceraldehyde 3-phosphate dehydrogenase (G3PDH), a key glycolytic enzyme. However, such severe cell death was effectively (approximately 85%) prevented with N-acetylcysteine (NAC), a precursor of reduced glutathione (GSH) that is an essential cofactor for Gly-I, accompanied by the intact Gly-I and G3PDH activities. Therefore, Gly-I may play a critical detoxifying role in glycolysis to maintain cellular activity and viability of prostatic cancer cells. PMID- 12086017 TI - Determination of pyruvate kinase type tumor M2 in human renal cell carcinoma: a suitable tumor marker? AB - Renal cell carcinoma (RCC) expresses an isoform of the glycolytic enzyme pyruvate kinase (type M2). The dimeric form (TuM2-PK) is over expressed in tumor cells and is detectable in blood with a sensitive enzyme-linked immunosorbent assay (ELISA). The aim of the present study was to evaluate the clinical value of TuM2 PK as a tumor marker for RCC. The TuM2-PK concentration in EDTA-plasma was determined quantitatively and immunologically using an ELISA. We measured the TuM2-PK plasma levels of 83 patients before and after surgery. Ninety-seven patients with various non-malignant diseases were also recruited as a control group. The control group displayed mean levels of 11.37 U/ml of TuM2-PK. Values were elevated in patients with RCC prior to surgery (mean 21.88 U/ml). The plasma levels increased after surgery until day 5 (mean 53.97 U/ml). At day 10, marker levels started to decrease without reaching preoperative values (mean 43.5 U/ml). Plasma levels in the renal vein (obtained during surgery) were not different from those in the peripheral blood. Follow-ups after 2-6 months showed a decrease to below preoperative levels (mean 16.3 U/ml). A significant difference was obtained by comparing the patients according to their Robson score. We found a significant difference (P < 0.01, Wilcoxon's two-sample test) in TuM2-PK levels between patients with RCC and the control group. Nevertheless, using the manufacturer's recommended cut-off value (15 U/ml), sensitivity was only 50.6% and specificity was 80.4%. Our results suggest that TuM2-PK is not a suitable tumor marker for RCC. PMID- 12086018 TI - TGF-beta1 in patients with renal cell carcinoma. AB - Up to now, clinical tumor-markers for renal cell carcinoma (RCC) have been lacking. Increased plasma levels of transforming growth factor-beta1 (TGF-beta1) were described as a tumor-marker and prognostic factor in RCC. The aim of this study was to test the clinical suitability of plasma TGF-beta1 as a tumor-marker for RCC. The concentrations of active and latent TGF-beta1 were determined in plasma of patients with localized (n = 39) and metastasised (n = 17) RCC. A newly developed, highly sensitive ELISA, which is specific for the isoform beta1, was used. Active TGF was directly measured in the EDTA plasma. To determine the amount of latent TGF-beta1, which is bound predominantly at beta2-macroglobulin, an optimized activation procedure was applied. Patients with localized RCC showed median concentrations of 16,700 ng/l (6,200-54,800 ng/l) for latent TGF-beta1. A total of 94 patients with various nonmalignant urological diseases were recruited as a control group. In comparison, this group had median concentrations of 19,900 ng/l (2,640-52,300 ng/l) for latent TGF-beta1. There was no significant difference (nonparametric Kruskal-Wallis ANOVA) between these groups. Patients with metastatic RCC showed median concentrations of 34,500 ng/l (6,800-48,960 ng/l) for latent TGF-beta1. In comparison to the localized RCC group, a statistically significant difference was found. Plasma levels after operative therapy (days 1, 5 and 10) and during follow-up without evidence of disease (2-6 months) showed no significant differences. Contrary to other study groups, our results suggest that TGF-beta1 is not a suitable tumor-marker for the diagnosis of localized RCC. In the face of higher TGF-beta1 plasma levels in metastatic disease, TGF-beta1 may be useful in the early detection of RCC recurrence or to control the success of immunochemotherapy. PMID- 12086019 TI - Estramustine phosphate enhances the effects of hyperthermia and induces the small heat shock protein HSP27 in the human prostate carcinoma cell line PC-3. AB - The antimicrotubule drug estramustine phosphate (EMP) has been shown to sensitize prostate carcinoma cells to radiation via synchronization at the G2/M phase of the cell cycle. This synchronization may also render cells more sensitive to hyperthermia, providing a rationale for multimodal treatment approaches. We have investigated the effects of EMP and hyperthermia, as well as the regulation of heat shock proteins (HSP) in the PC-3 prostatic carcinoma cell line. Cells were incubated with four doses of EMP for 48 h followed by a 1-h hyperthermia treatment ranging from 41 degrees C to 44 degrees C. Cell cycle distribution at the end of the EMP incubation was investigated by flow cytometry. Cytotoxicity was assessed by colony formation assays. HSP accumulation was investigated by Western immunoblotting. Doses of 1, 5, 10 and 15 microM EMP synchronized 27, 28, 46, and 68% of PC-3 cells at G2/M. With 5, 10 and 15 microM, a sensitizing effect of EMP was assessed at hyperthermic temperatures of 42, 43 and 44 degrees C. EMP did not alter the expression of HSP72, but substantially induced the synthesis of HSP27 in PC-3 cells. Our data show that EMP sensitizes PC-3 cells to hyperthermia induced cytotoxicity. This observation supports the rationale for multimodal treatment approaches in locally advanced prostate cancer. PMID- 12086020 TI - The rise and fall of PSA: clinical implications of prostate specific antigen kinetics. PMID- 12086021 TI - Impact of different combined preservation modalities on warm ischemic kidneys: effect on oxidative stress, hydrostatic perfusion characteristics and tissue damage. AB - Hemodynamic disorders in brain dead organ donors induce hypoxia, warm ischemia and finally tissue damage. A cold preservation period also induces tissue and cellular lesions. The two major modes of preservation are cold storage (CS) and hypothermic pulsatile perfusion (HPP). We aimed to compare the influence of each mode of preservation and their combination on oxidative stress, perfusion characteristics and tissue damage, after a period of warm ischemia. Rat kidneys which had undergone ischemia (0, 30, 60 min) were preserved either by CS (12, 24 h), or by HPP (12 h), or by a combination of both (HPP+CS, CS+HPP), in University of Wisconsin cold storage solution (UWCSS) at + 4 degrees C. During HPP, renal vascular pressure decreased then increased to reach 90 mmHg after perfusion for 7 h. If HPP followed CS, the mean pressure reached 200 mmHg, showing successive high amplitude peaks. HPP had a deleterious effects on tissue structure with tubular necrosis, and induced an increase in catalase (Cat) and a decrease in manganese superoxide dismutase (Mn SOD) and gluthatione peroxidase (GPx) activity. Copper zinc superoxide dismutase (Cu/Zn SOD) activity was not reduced except with CS+HPP. During CS, we observed an increase in GPx, Cu/Zn SOD and Cat activity, a decrease in Mn SOD activity and no histological alterations in the kidney. CS induces a slight oxidative stress which is not important enough to induce major tissue damage. HPP with UWCSS induces a stronger stress, which overpowers the antioxidant defences, inducing tissue damage. The reperfusion of HPP with UWCSS emphasises the stress initiated by CS. In addition an increase in damage occurred in the CS + HPP group. PMID- 12086022 TI - Local renal and vascular natriuretic peptide system in obstructive uropathic rats. AB - The present study was aimed at investigating whether the regulation of the local natriuretic peptide system is altered in the kidney and the vasculature in obstructive uropathy. Male Sprague-Dawley rats were bilaterally obstructed by ligation of the proximal ureters for 48 h. Control rats were treated in the same way, except that no ligature was made. The mRNA expression of the various isoforms of atrial, brain, and C-type natriuretic peptide (ANP, BNP, CNP) and different subtypes of natriuretic peptide receptor-A, -B, and -C (NPR-A, NPR-B, NPR-C) was determined in the kidney and the thoracic aorta by reverse transcription-polymerase chain reaction. The basal and stimulated activities of particulate guanylyl cyclase were also examined. Following the bilateral ureteral obstruction, the expression of ANP, BNP, and CNP was increased in the aorta as well as in the kidney. Contrary to this, the expression of NPR-A, NPR-B, and NPR C was decreased both in the kidney and the aorta. Accordingly, the guanylyl cyclase activity was significantly decreased in response to natriuretic peptides. ANP relaxed phenylephrine-precontracted aortic rings in a dose-dependent manner, the degree of which was significantly diminished. Our results suggest that the local synthesis of natriuretic peptides is increased in the kidney and in the vasculature in obstructive uropathy. PMID- 12086023 TI - Lessons from hot feet: a note on tissue remodeling. PMID- 12086024 TI - Outcomes, treatment effectiveness, efficacy, and evidence-based practice: examples from the world of splinting. AB - Calls for outcome studies and evidence-based practice leave many therapists bewildered as to the meaning of various types of terminology. The objectives of this paper are to 1) review the structure of outcome reports and research; 2) describe the background and discuss the terms "treatment efficacy, "effectiveness," and "evidence-based practice"; 3) discuss these concepts as they apply to selected studies found in the literature that report the use of splints for patients with ailments that affect the use of their hands; and 4) provide some reflections that place these terms in the context of scientific inquiry and clinical practice. It is concluded that health care is being reviewed on an increasingly more systematic and grand scale. While studies that include large numbers of subjects are needed to substantiate the effectiveness of splinting, studies involving smaller numbers of subjects can contribute greatly as well, so long as they are performed with the highest level of study design and use standardized outcome measures that are valid for answering the designated research questions. PMID- 12086025 TI - Plaster of Paris: the forgotten hand splinting material. AB - This article examines the concept of tissue adaptation in response to the application of plaster of Paris splints and casts. A review of the history of plaster of Paris and its composition, its working properties, and precautions for its use introduces the reader to this oft-forgotten material. Four designs are described for plaster of Paris application-circumferential padded casts, digital unpadded casts, plaster slabs, and contour molds. The discussion of clinical application of plaster of Paris covers joint tightness, arthritis, contracted joints due to spasticity, muscle-tendon tightness, skin tightness, skin and joint tightness, and edema reduction. In addition, a new application called casting motion to mobilize stiffness (CMMS), developed by the author, is discussed. The use of plaster of Paris to improve postoperative flexor tendon glide is also discussed. This review article intends to stimulate the reader to use plaster of Paris splinting or casting more frequently to solve clinical problems. PMID- 12086026 TI - A proposed decision hierarchy for splinting the stiff joint, with an emphasis on force application parameters. PMID- 12086027 TI - Static progressive splinting. AB - Static progressive splinting is the use of inelastic components to apply torque to a joint in order to statically position it as close to end range as possible. It maximizes total end-range time, thus increasing passive range of motion. As tissue lengthens in response to this carefully applied stress, the clinician or wearer adjusts the joint position to progress tissue at the new maximum tolerable length. Static progressive splinting combines precision in joint position and torque application with patient-controlled stress to create an approach powerful enough to succeed when no other treatment approach does. This article discusses static progressive splinting indications, contraindications, and advantages as well as guidelines for a splinting regimen. It offers many examples of static progressive splinting and makes clear that this approach can be used with any mobilizing splint design. The unique mechanics of this splinting approach are described, and the various methods of achieving static progressive splinting are compared. Offering high levels of patient satisfaction and compliance, static progressive splinting has come to the forefront of clinical practice. PMID- 12086028 TI - The effect of the forearm support band on forces at the origin of the extensor carpi radialis brevis: a cadaveric study and review of literature. AB - The forearm support band is hypothesized to reduce force, and thus inflammation, at the extensor carpi radialis brevis (ECRB) origin. Very little published evidence supports an actual effect on the forces at the ECRB origin, however. The authors describe a cadaveric study in which forces at the ECRB origin were measured while various pressures were applied to the forearm support band and the ECRB tendon was loaded distally. The results reveal an increased effect with increased band pressure and a decreased relative effect with increased force applied distally. For clinical correlation, application pressure was also measured in nine patients using a counterforce brace. While further clinical evaluation is necessary to determine the most appropriate clinical indications for use of a forearm support band, these results suggest that the forearm support band may be most effective when applied at a force of 40 to 50 mmHg and used during light-duty activities. PMID- 12086029 TI - The effect of corrective splinting on flexion contracture of rheumatoid fingers. AB - This paper reports a matched-pair experimental study to investigate the effect of corrective splinting on flexion contracture of rheumatoid fingers. Twenty-four patients with rheumatoid arthritis and finger flexion contracture participated in the study. After a 6-week baseline measurement of hand function-including measurement of grip strength and range of motion and administration of the Jebsen Hand Function Test-the patients were randomly placed into two groups. Patients in the first group were given dynamic (Capener) splints, and those in the second group were given static (belly gutter) splints. Hand function was re-assessed 6 weeks after the splinting program. Results indicated significant improvement in both groups, not only in the correction of the finger flexion contracture (p < 0.0005) but also in grip strength (p = 0.001) and hand function (p < 0.0005). Patients with dynamic finger extension splints did not differ from those with static splints in extension gains, but they did have better flexion than patients with static splints. Both types of splints can be recommended for flexion contracture of rheumatoid fingers, depending on patients' preferences and comfort. PMID- 12086030 TI - Minimalistic splint design: a rationale told in a personal style. AB - Theoretic clinical reasoning is becoming increasingly important in splint construction. The purpose of this paper is to present an algorithm of decision steps that will help therapists consider all aspects of splint making proactively, so that they can take the necessary steps to be successful. A decision flowchart is presented and discussed in detail. An extra dimension of splint design is emphasized-namely, frugality in structure, technique, and cost. The paper concludes that the use of a comprehensive decision algorithm provides the basis for successful working technique and that the ability to create minimal design splints is more a state of mind than a technical skill. PMID- 12086031 TI - Origins and application of bark cloth in upper extremity splinting. PMID- 12086032 TI - Introduction to a new method for inelastic mobilization. PMID- 12086033 TI - The splinting issue. PMID- 12086034 TI - A history of splinting: to understand the present, view the past. AB - The purpose of this study was to identify, using an intensive literature review, the primary historical factors and events that shaped the evolution of current splinting technique and practice. Publication dates determine the chronological order of events, and splints are defined according to the ASHT Splint Classification System. Purposes of splinting are analyzed and listed according to frequency of citation in splint manuals and books, and a historical review of splinting practice describes splints, events, and persons who influenced the evolution of splinting from the time of Hippocrates through the 20th century. Factors influencing splint development include disease, political conflict, advancements in medicine and technology, agency and organizational decision making, centers of practice, and availability of information. Infection, polio, war, technology, plastics, surgical advances, soft tissue remodeling, anatomy, biomechanics, government agencies, hand centers, seminars, professional organizations, publications, and a classification system have all played important roles in 20th century splinting practice. PMID- 12086035 TI - Determination of shelf life of concentrated yogurt (labneh) produced by in-bag straining of set yogurt using hazard analysis. AB - Strained yogurt, labneh, produced by straining cow's milk set yogurt in cloth bags, was stored at 5, 15, and 25 degrees C, and changes in microbial counts, pH, titratable acidity, percentage of free whey, and sensory attributes were monitored during storage. Counts of total aerobes, psychrotrophic yeasts, yeasts and molds, and lactic acid bacteria, except in samples stored at 25 degrees C, increased irrespective of storage temperature. The pH of samples decreased, titratable acidity and percentage of free whey increased, and texture defects were detected at a later stage than flavor changes during storage. Shelf-life data of labneh was adequately described by the Weibull distribution. The nominal shelf life determined using sensory changes and yeast counts as failure criteria ranged from 8.5 to 10.5, 4.7 to 5.8 and 2.3 to 2.7 d at 5, 15 and 25 degrees C, respectively. Q10 (shelf life at T degrees C/shelf life at T+10 degrees C) for flavor quality loss was 1.98 at 5 degrees C, and the corresponding activation energy was 11.3 kcal/mol. PMID- 12086036 TI - Influence of a Spirulina platensis biomass on the microflora of fermented ABT milks during storage (R1). AB - The objective of this research was to investigate the effect of a cyanobacterial (Spirulina platensis) biomass on the microflora of a probiotic fermented dairy product during storage at two temperatures. Spirulina-enriched and control (plain) fermented acidophilus-bifidus-thermophilus (ABT) milks were produced using a fast fermentation starter culture (ABT-4) as the source of Lactobacillus acidophilus (A), bifidobacteria (B), and Streptococcus thermophilus (T). Incubation took 6 h at 40 degrees C. As for the cyanobacterial product, the S. platensis biomass was added to the process milk during stirring at pH 4.5 to 4.6. Thereafter, the ABT-type fermented milks were cooled to 25 degrees C in ice water, filled into sterile, tightly capped centrifuge tubes, further cooled at 4 degrees C for 24 h, and then stored either at 15 degrees C for 18 d or at 4 degrees C for 42 d. Microbiological analyses and acidity measurements were performed at regular intervals. Our results showed that the counts of the starter organisms were satisfactory during the entire storage period at both temperatures applied in this research. The S. platensis biomass had a beneficial effect on the survival of ABT starter bacteria regardless of storage temperature. Postacidification was observed at 15 degrees C, whereas pH remained stable during refrigerated storage at 4 degrees C. The abundance of bioactive substances in S. platensis is of great importance from a nutritional point of view because thus the cyanobacterial biomass provides a new opportunity for the manufacture of functional dairy foods. PMID- 12086037 TI - Species-specific identification of commercial probiotic strains. AB - Products containing probiotic bacteria are gaining popularity, increasing the importance of their accurate speciation. Unfortunately, studies have suggested that improper labeling of probiotic species is common in commercial products. Species identification of a bank of commercial probiotic strains was attempted using partial 16S rDNA sequencing, carbohydrate fermentation analysis, and cellular fatty acid methyl ester analysis. Results from partial 16S rDNA sequencing indicated discrepancies between species designations for 26 out of 58 strains tested, including two ATCC Lactobacillus strains. When considering only the commercial strains obtained directly from the manufacturers, 14 of 29 strains carried species designations different from those obtained by partial 16S rDNA sequencing. Strains from six commercial products were species not listed on the label. The discrepancies mainly occurred in Lactobacillus acidophilus and Lactobacillus casei groups. Carbohydrate fermentation analysis was not sensitive enough to identify species within the L. acidophilus group. Fatty acid methyl ester analysis was found to be variable and inaccurate and is not recommended to identify probiotic lactobacilli. PMID- 12086038 TI - Application of IS900 PCR for detection of Mycobacterium avium subsp. paratuberculosis directly from raw milk. AB - A polymerase chain reaction (PCR)-based assay was developed for detection of insertion sequence 900 (IS900) of Mycobacterium avium subsp. paratuberculosis in raw milk. This IS900 PCR assay included DNA extraction and PCR assay using commercially available kits. The DNA extraction and PCR assay were optimized to detect the IS900 sequence directly from raw milk. The IS900 PCR assay was evaluated by inoculating raw bulk milk and Middlebrook's 7H9 broth with 0 to 10(8) cfu/ml of each of four American Type Culture Collection strains of M. paratuberculosis. Under experimental conditions, both milk culture on Herrold's egg yolk medium slants, and IS900 PCR could detect 10 to 100 cfu/ml of M. paratuberculosis. Detection of M. paratuberculosis by IS900 PCR was consistent (24/24 PCR assays) when about 100 cfu/ml were present, whereas detection was variable (12/24 PCR assays) at concentrations as low as 10 cfu/ml. Based on the findings of the experimental study, IS900 PCR was further evaluated with pooled quarter milk samples from 211 cows from five herds with known history of Johne's disease. Out of 211 animals examined, nine (4%) and 69 (33%) were positive for M. paratuberculosis by milk culture and IS900 PCR from milk, respectively. A total of 20 bulk tank milk sample aliquots (one sample, four aliquots from each herd) were also examined, of which 10 (50%) were positive for M. paratuberculosis by IS900 PCR. By contrast, only one out of 20 (5%) bulk tank milk sample aliquots was positive by culture. The IS900 PCR amplified product of 229-bp obtained on testing of quarter milk and bulk tank milk samples was confirmed to be the IS900 of M. paratuberculosis by DNA sequence analysis. The results of this study suggest that M. paratuberculosis can be detected directly from quarter milk and bulk tank milk by IS900 PCR. PMID- 12086039 TI - Production of oligosaccharides in yogurt containing bifidobacteria and yogurt cultures. AB - Yogurts were prepared by using yogurt cultures combined to mixed cultures of bifidobacteria (Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, and Bifidobacterium longum) and by adding a preincubation step (1.5 h at 50 degrees C) with bifidobacteria to the conventional method of manufacture in order to produce oligosaccharides. The survival of bifidobacteria was drastically affected during storage of yogurts, except for products containing B. animalis, in which viable counts remained at >10(6) cfu/g after 28 d of storage at 4 degrees C. Oligosaccharides with a degree of polymerization of 3 were produced during the preincubation step (0.31 to 0.68%), and the amount in the final products varied according to the species of bifidobacteria inoculated during the preincubation step or the concentration of bifidobacteria used as second inoculum during the fermentation process. In fact, the higher concentration of oligosaccharides measured at the end of the fermentation process (0.72%) and the 28 d-storage period (0.67%) was obtained for yogurts containing B. infantis. However, yogurts containing B. breve showed higher beta-galactosidase activities and had lower lactose concentrations after the fermentation process and the storage period than the other yogurts. The use of a mixed cultures of bifidobacteria (B. animalis, B. infantis, or B. breve) thus allows the production of yogurts in which bifidobacteria can survive in relatively high cell numbers and contain appreciable amount of oligosaccharides. PMID- 12086040 TI - Effects of added plasmin on the formation and rheological properties of rennet induced skim milk gels. AB - Elevated plasmin enzyme activity has been suggested as a likely cause of impaired functional properties that occur in milk from cows either in their late lactational period or that are experiencing mastitis. However, there are conflicting reports on the impact of plasmin on rennet coagulation properties of milk. The effects of added plasmin on the rheological properties, at small and large deformation, of rennet-induced gels were investigated. The microstructure of rennet-induced gels was studied, using confocal scanning laser microscopy. Porcine plasmin was added to reconstituted milk, and samples were incubated at 37 degrees C for between 0.5 to 8 h. The hydrolysis reaction was terminated using soybean trypsin inhibitor. The extent of degradation of caseins was determined with SDS-PAGE. The extent of breakdown of alpha(s)- and beta-caseins increased with incubation time with plasmin. Storage modulus of rennet gels decreased linearly with increasing degradation of caseins. There was an increase in the loss tangent parameter of the gels with increasing casein degradation, indicating a more liquid-like gel character. Gelation time decreased until approximately 3 h of incubation with plasmin (when the amounts of intact alpha(s)- and beta-caseins were approximately 46 and 50%, respectively); thereafter, gelation time increased considerably. Yield stress of rennet-induced gels decreased with increasing casein breakdown. When the level of casein hydrolysis was high (<40% of intact caseins), the microstructure of rennet-induced gels was drastically altered. Even when there were low levels of casein hydrolysis, the rheological properties of rennet gels were altered, which could have negative impacts on cheese yield and texture. PMID- 12086041 TI - Milk protein response to abomasal or mesenteric vein essential amino acid infusion in lactating dairy cows. AB - Previous experiments performed at our location suggested that the milk protein response to infusions of mixed essential amino acids (EAA) was greater when these were supplied via intravenous rather than abomasal or duodenal routes. However, as far as we are aware there have been no direct comparisons of the milk protein response to site of EAA provision in the same animals. Our objective was to directly compare the milk protein responses when cows were given mixtures of EAA provided via abomasal or mesenteric vein infusions. Four multiparous, ruminally cannulated, multicatheterized Holstein x Friesian cows averaging 18 wk postpartum were fed dehydrated alfalfa, grass silage, and low protein (11.9% CP, DM basis) concentrates at 30, 20, and 50%, respectively, of dry matter (DM) offered. Total mixed rations (14% crude protein, DM basis) were fed daily as 3 equal meals at 8 h intervals and 95% of ad libitum DM intake. The experimental design was a single reversal with two 10-d infusion periods separated by a 10-d rest period. Each infusion period consisted of 4-d control infusions into the mesenteric vein (saline at 2 ml/min) and abomasum (water at 9 L/d) followed by 6-d infusion of a mixture of EAA equal to 400 g of milk protein/d into the abomasum or mesenteric vein. Control infusions continued into the site not receiving EAA. Intake of DM (20.3 kg/d), milk yield (28.9 kg/d), and milk fat concentration (41.2 g/kg) were not affected by EAA infusions. Milk protein concentration (33.4 vs 34.6 g/kg) and output (938 vs. 982 g/d) were increased and milk lactose concentration was decreased (46.6 vs. 46.1 g/kg) by EAA, but the responses were not affected by infusion site. Recovery of EAA as increased milk protein output (10.9%) was similar for intravascular and abomasally infused EAA in these cows, but these responses were low compared with published effects of postruminal casein infusion. PMID- 12086042 TI - Metabolic responses of transition Holstein cows fed anionic salts and supplemented at calving with calcium and energy. AB - The objective of this study was to determine the concentrations of plasma Ca, P, Mg, nonesterified fatty acids (NEFA), 3-hydroxybutyrate (BHBA), and glucose in transition cows fed anionic salts prepartum and provided with calcium and energy supplements at calving. The study was conducted on a Florida Holstein dairy farm from November to December 1997. Treatments consisted of no treatment (n = 30); 60 g of Ca as calcium chloride, orally (n = 30); 110 g of Ca as calcium propionate 510 g plus 400 g of propylene glycol, orally (n = 30); two doses of 60 g of Ca as calcium chloride, one at calving and the second 24 h later, orally (n = 30); and 10 g of Ca as borogluconate, intravenously (n = 30). Treatments were administered within 12 h after parturition. Blood samples were collected at d 1 (parturition), 2, 3, 6, 9, and 12 after calving. Plasma total Ca, P, Mg, NEFA, BHBA, and glucose were measured. There were no differences in the concentrations of the blood metabolites among treatments. PMID- 12086043 TI - Bovine somatotropin administration to dairy goats in late lactation: effects on mammary gland function, composition and morphology. AB - We investigated the effects of bovine somatotropin (bST) on mammary gland function and composition in the declining phase of lactation in goats. Sixteen Saanen goats, 180 +/- 11 days in milk (DIM), were divided equally into control and treated groups. The treated group received 120 mg/2 wk of slow-release bST for three cycles. Milk yield, milk composition, milk clotting measures, and plasmin-plasminogen activator activities were recorded weekly. Milk Na and K were determined in individual milk samples collected weekly during the third cycle. Blood samples were collected weekly during the second cycle and the plasma analyzed for nonesterified fatty acids (NEFA), glucose, and urea. At the end of the 6 wk, three goats from each group were slaughtered, and the udders were removed. Mammary gland weight, composition, and total DNA content were determined. The histological effects of bST on mammary tissue were investigated. The analyzed parameters included numbers of alveoli, corpora amylacea, apoptotic cells, and laminin fibronectin distribution and localization. An extensive morphological analysis on the epithelial and stromal components was performed. Milk yield was significantly higher in the treated group, fat content was not affected, but protein and nonprotein nitrogen were lower in treated goats milk. Treatment with bST did not influence milk pH but reduced coagulation time. Plasmin and plasminogen activator activities were not affected. Milk K levels were higher and the Na/K ratio was lower in treated animals. Plasma glucose, NEFA, and urea were unaffected. Mammary gland weight and total DNA were higher in treated than control animals, suggesting that with advancing lactation bST treatment maintains cells. Fat, protein, and collagen content of the mammary tissue did not differ between the groups. Treatment with bST significantly increased the number of lactating alveoli (LA) and significantly reduced the number of regressing alveoli (RA) and corpora amylacea, both within and outside the alveolar lumen. Laminin and fibronectin localization were not affected, and very few apoptotic cells were found in both treated and control samples. Our findings suggest that bST administration to dairy goats in late lactation can modulate mammary gland activity and improve lactation persistency; this is associated with maintained total mammary parenchyma weight and lactating alveoli. PMID- 12086044 TI - Regulation of apoptosis during mammary involution by the p53 tumor suppressor gene. AB - Regulation and functions of the p53 tumor suppressor gene have been studied extensively with respect to its critical role in maintaining the stability of genomic DNA following genotoxic insults. However, p53 is also induced by physiologic stimuli resulting in cell cycle arrest and apoptosis. In other situations, the activity of p53 must be repressed to prevent inappropriate removal of cells. The mammary gland provides a valuable system in which to study the mechanisms by which the expression and biological responses to p53 can be regulated under a variety of physiological circumstances. The pro-apoptotic role of p53 in the secretory mammary epithelium may be especially relevant to lactation in livestock. We have utilized p53-deficient mice to establish the molecular targets of p53 in the mammary gland and biological consequences when it is absent. The p21/WAF1 gene (Cdkn1a) is a transcriptional target gene of the p53 protein that responds to elevated levels of p53 during milk stasis providing an endogenous reporter of p53 activity. Abrogation of p53 resulted in delayed involution of the mammary epithelium, demonstrating the physiological role of p53 in regulating involution. Though delayed, stromal proteases were induced in the mammary gland by 5 d postweaning, providing a p53-independent mechanism that resulted in removal of the residual secretory epithelium. These processes can be interrupted by treatment with hydrocortisone. These data establish p53 as a physiological regulator of involution that acts to rapidly initiate apoptosis in the secretory epithelium in response to stress signals, but also indicate the presence of compensatory pathways to effect involution. Additional mechanisms involving intracellular stress signaling pathways (e.g., Stat3) and stromal mediated pathways have been identified and, together with p53 pathways, may be used to identify animals with greater persistency of lactation. PMID- 12086045 TI - Trends in antibacterial susceptibility of mastitis pathogens during a seven-year period. AB - Milk samples collected from dairy cattle suspected of having mastitis were submitted to the Microbiology Laboratory of the Animal Health Diagnostic Laboratory, Michigan State University, for bacteriologic culture. A total of 2778 isolates, from the years 1994 to 2000, were isolated, identified, and subjected to in vitro antimicrobial susceptibility testing using the disk diffusion method, in accordance with National Committee on Clinical Laboratory Standards (NCCLS) standards. Isolates included in this study were Streptococcus uberis, Streptococcus dysgalactiae, Streptococcus agalactiae, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Serratia marcesens, and Pseudomonas aeruginosa. The proportion of bacterial isolates determined to be susceptible did not change during the 7-yr period for the majority of bacterial-antibacterial interactions tested. However, analysis for linear trend in proportions determined that there were increases in the proportion of S. aureus isolates that were susceptible to ampicillin, penicillin, and erythromycin. For Strep. uberis, increases in the proportion of susceptible isolates occurred for oxacillin, sulfa trimethoprim, gentamicin, and pirlimycin, and a decrease in the proportion of susceptible isolates occurred with penicillin. For Strep. dysgalactiae, increases in the proportion of susceptible isolates occurred with erythromycin, gentamicin, sulfa-trimethoprim, and tetracycline. For Strep. agalactiae, increases in the proportion of susceptible isolates occurred with sulfa-trimethoprim. Among E. coli isolates, there was an increase in the proportion that were susceptible to ampicillin and cephalothin. Among K pneumoniae isolates, there was an increase in the proportion that were susceptible to ceftiofur. Overall, there was no indication of increased resistance of mastitis isolates to antibacterials that are commonly used in dairy cattle. PMID- 12086046 TI - Monitoring nonlactating cow intramammary infection dynamics using DHI somatic cell count data. AB - Although the nonlactating period presents a risk for intramammary infection, efficient systems to monitor infection status of recently calved cows have not been developed, and benchmarks for interpretation have not been established. Individual cow somatic cell count (SCC) data for the current and previous six monthly Dairy Herd Improvement milk tests and the last SCC of the previous lactation and first SCC of the current lactation were summarized for all milking cows in a selection of Wisconsin dairy herds. Prevalence of infection, herd new infection rate, fresh cow contribution to herd new infection rate, dry cow new infection rate, heifer new infection rate, and dry cow cure rate were estimated using a threshold of 200,000/ml. In 145 herds, mean (range) heifer new infection rate was 21.3% (0 to 58%). The cut-point for the 10th percentile of herds was 8%. Mean (range) dry cow new infection rate in cows that were uninfected at the last test before dry off was 22.4% (0 to 71%), and the cut-point for the 10th percentile of herds was 9%. Although nonlactating cow and heifer new infection rates increased with weighted 6-mo mean herd SCC, the between-herd variation was large, suggesting that on-farm factors are important in determining the rates of infection. In a subset of 51 Wisconsin dairy herds, significant monthly variation in weighted SCC, prevalence, herd new infection rate, and fresh cow contribution to herd new infection rate were detected. Elevations in SCC and prevalence of infection during the summer (July through September) were associated with significant increases in fresh cow and herd new infection rates. PMID- 12086048 TI - Influence of Staphylococcus aureus strain-type on mammary quarter milk somatic cell count and N-acetyl-beta-D-glucosaminidase activity in cattle from eight dairies. AB - The hypothesis tested was that there are differences in pathogenicity between strains of Staphylococcus aureus that cause bovine mastitis. Mammary quarter milk somatic cell count (SCC) and N-acetyl-beta-D-glucosaminidase (NAGase) activity were used as indicators of the pathogenicity of different strains of S. aureus that infect the bovine udder. Eight commercial dairy herds with a history of S. aureus in bulk tank milk cultures were studied. Initially, composite foremilk samples were collected from all lactating cattle in each herd and cultured for staphylococci. Subsequently, all cows with a coagulase-positive staphylococcal intramammary infection (IMI) at the initial sampling that were still present in the herd of origin had individual mammary quarter foremilk samples collected. Coagulase-positive staphylococcal isolates were confirmed as S. aureus using a commercial biotyping system. Staphylococcus aureus isolates were strain-typed using pulsed-field gel electrophoresis. Mammary quarter milk SCC and N-acetyl beta-D-glucosaminidase activity were determined for each cow. The difference in mean somatic cell count and mean NAGase activity for mammary quarters infected with the same strain of S. aureus and for uninfected quarters on the same cow was calculated. One-way analysis of variance was used to assess differences between strains within a herd. Overall, no significant differences were found between strains, suggesting that the degree of udder parenchymal injury induced by S. aureus IMI is in general significantly affected by factors other than strain type. PMID- 12086047 TI - Evaluation of five selective media for isolation of catalase-negative gram positive cocci from bulk tank milk. AB - Five selective media including Edwards modified medium, Edwards modified medium supplemented with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L), Streptococcus selective medium, Streptosel agar, and thallium-crystal violet toxin-ferric citrate medium were evaluated for the isolation of streptococci and streptococci-like organisms from raw milk. The sensitivity and specificity of these selective media for streptococci and streptococci-like organisms were determined by using American Type Culture Collection reference strains. Under experimental conditions Edwards modified medium with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L) showed the highest sensitivity (100%) and specificity (100%) for streptococci and streptococci-like organisms followed by thallium-crystal violettoxin-ferric citrate medium, Edwards modified medium, Streptococcus selective medium, and Streptosel agar. Edwards modified medium supplemented with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L) allowed growth of all streptococci and streptococci-like organisms, while inhibiting growth of the staphylococci and gram-negative reference strains. Bulk tank milk samples from 114 dairy herds were spiral plated onto Edwards modified medium with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L). A total of 344 isolates (at least three isolates from each sample) were randomly selected and identified to their species. This medium permitted growth of 328 streptococci and streptococci-like organisms belonging to genera Aerococcus, Enterococcus, Gemella, Lactococcus, Streptococcus, and Vagococcus. When Edwards modified medium supplemented with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L) was evaluated using bulk tank milk samples, the sensitivity and specificity of this medium for streptococci and streptococci-like organisms were observed to be 100 and 87.5%, respectively. The positive predictive value for streptococci and streptococci-like organisms was observed to be 99.4%. The results of the study indicate that Edwards modified medium supplemented with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L) can be used as a selective medium for the isolation of streptococci and streptococci-like organisms from bulk tank milk. PMID- 12086049 TI - Effect of lactoferrin in combination with penicillin on the morphology and the physiology of Staphylococcus aureus isolated from bovine mastitis. AB - The objective of the present study was to evaluate the therapeutic potential of bovine lactoferrin or lactoferricin in combination with penicillin G against Staphylococcus aureus. Minimal inhibitory concentrations of lactoferrin, lactoferricin, penicillin, and combinations of lactoferrin or lactoferricin with penicillin were determined for 15 S. aureus strains including several strains resistant to beta-lactam antibiotics. The fractional inhibitory concentration index indicated a synergistic effect between lactoferrin and penicillin. Combination of lactoferrin with penicillin increased the inhibitory activity of penicillin by two- to fourfold and reduced the growth rate in S. aureus strains tested, whereas the increase in the inhibitory activity of lactoferrin by penicillin was 16- to 64-fold. The addition of iron to the medium containing a combination of penicillin and lactoferrin had no effect on growth inhibition. Electron microscopy revealed that concentration below the minimal inhibitory concentrations of penicillin induced important ultrastructure alterations, which were further enhanced by the presence of lactoferrin. When S. aureus cells were grown in the presence of a combination of penicillin and lactoferrin, changes in the protein profile of the bacteria, including the disappearance of several protein bands due to the presence of lactoferrin, were observed. These data suggest that bovine lactoferrin or lactoferricin in combination with beta-lactam antibiotics can increase the antibacterial activity of these antibiotics against S. aureus resistant to antibiotics. PMID- 12086050 TI - The relationship between estrous behavioral score and time of ovulation in dairy cattle. AB - One of the major contributors to poor fertility of a dairy herd is ineffective detection of estrus. Recently, it has become evident that cow factors contribute largely to low detection rates. Until now, standing behavior has been the symptom used to determine the right moment for insemination. However, standing behavior is not observed in over 50% of the cows in estrus in a number of herds, and no data are available to relate the moment of ovulation to other estrous symptoms. Therefore, in the present study, cows were observed for a number of visual signs of estrus, twice per day for 30 min, and the moment of ovulation was estimated by transrectal ultrasonographic scanning of the ovaries. In this study, 100 cows were detected in estrus visually, using a scoring system. Standing behavior was observed in 50% of those cows, although there was more than one cow in estrus at a time in most (85%) cases. There was no correlation between follicular size and ovulation time or estrous behavior score. Levels of 305-d milk yield and parity were also not correlated with estrous behavior scores. However, a significant correlation (0.31) was found between detection score and day of ovulation. A retrospective analysis revealed that cows that ovulated 0 to 24 h after artificial insemination (AD) scored almost three times the number of estrous behavior points compared to the ones that ovulated 24 to 48 h after AI. Ovulation more than 48 h after AI resulted in pregnancy in only 15% of the cows. PMID- 12086051 TI - The effect of left displacement of abomasum corrected by toggle-pin suture on lactation, reproduction, and health of Holstein dairy cows. AB - Our objectives were to evaluate the effect of left displacement of abomasum (LDA) after correction by toggle-pin suture (TPS) on lactation performance, reproduction and health in Holstein dairy cows in a commercial dairy farm. Cows diagnosed with LDA and corrected by the TPS procedure (188 cows) during the first 70 d postpartum were matched with control herd-mates (186 controls) according to lactation number, calving date, and previous lactation 305-d mature equivalent milk yield. Cows were grouped according to parity and days in milk and fed the same total mixed ration throughout a 321-d lactation. Data collected included yields of milk and 3.5% fat-corrected milk (FCM), concentration and yields of milk fat, somatic cell count, incidence of mastitis, abortion, death and culling, in addition to reproductive measures. Cows affected with LDA corrected by the TPS procedure produced less milk and tended to produce less 3.5% FCM than control cows, but the decrease in production occurred only during the first 4 mo of lactation. Left displacement of abomasum did not affect the interval from calving to conception and conception rates, but it extended the period from calving to first postpartum artificial insemination. Incidences of abortions and mastitis were not influenced by LDA. Cows affected with LDA remained in the study for a shorter period than their control herdmates, and higher proportions of cows with LDA were sold or died. Death and culling were more pronounced immediately after the diagnosis of LDA and the TPS procedure. PMID- 12086052 TI - Effect of concentrate level and feeding management on chewing activities, saliva production, and ruminal pH of lactating dairy cows. AB - Eight ruminally cannulated lactating Holstein cows were used in a double 4 x 4 Latin square to determine the effects of 1) proportion of barley silage [40, 50, and 60% of dry matter (DM)] in the diet, and 2) feeding a total mixed ration (TMR) compared with separate ingredients (SI) on chewing activities, saliva production, and ruminal pH. Although cows fed SI were offered a diet containing 50% silage, they actually consumed a diet containing 43% silage (DM basis). Dry matter intake and milk yield were similar for all diets (18.2 kg of DM/d and 27.2 kg/d, respectively). Cows fed the 40% silage TMR spent more time eating than cows fed SI (243 vs. 198 min/d), but rumination time was similar (546 min/d). Eating time was similar among the TMR diets, but rumination time increased from 498 to 516 and 584 min/d as silage in the TMR increased from 40 to 50, and then to 60%, respectively. The secretion of saliva per gram of feed was 4.43, 3.18, and 1.19 ml/g of DM with consumption of silage, TMR, and concentrate, respectively. Resting salivation rate was similar for all diets (101 ml/min). Regardless of the diet, cows secreted 239 +/- 17 L/d of saliva, and ruminal pH was below 5.8 for 10 h/d. Results indicated increased chewing time did not increase total daily saliva secretion because increased eating and ruminating saliva was associated with decreased resting saliva. Feeding SI increased the risk of acidosis, because cows ate a higher proportion of concentrate than intended. PMID- 12086053 TI - Chewing activity, saliva production, and ruminal pH of primiparous and multiparous lactating dairy cows. AB - Four multiparous (MP) and four primiparous (PP) ruminally cannulated lactating Holstein cows were used in a double 4 x 4 Latin square design to study the chewing behavior, saliva production, and ruminal pH of cows in the first or subsequent lactation. Cows were fed one of four diets; three total mixed rations containing 40, 50, or 60% silage (DM basis), and a separate ingredient diet containing 50% concentrate. Dry matter intake was higher for MP cows than for PP cows (19.2 vs. 17.1 kg/d) but not as a percentage of body weight (2.97 +/- 0.06%). Multiparous cows spent more time eating than PP cows (260 vs. 213 min/d, respectively), even after adjustment for dry matter intake (13.8 vs. 12.4 min/kg DM). Multiparous cows also spent more time ruminating per day than PP cows (560 vs. 508 min/d, respectively). Eating salivation rate was not affected by parity, but resting salivation rate was higher for MP cows than for PP. Although MP cows spent more time chewing than PP cows, total daily saliva production was only numerically higher for MP cows because the increase in saliva produced during chewing was accompanied by a decrease in saliva produced during resting. Furthermore, pH profiles tended to be lower for MP cows than for PP cows. Multiparous cows may have a greater risk of incurring acidosis than PP cows because increased salivary secretion associated with increased chewing may not sufficiently compensate the increment of fermentation acids produced in the rumen due to high feed intake. PMID- 12086055 TI - Bovine milk fatty acid profiles produced by feeding diets containing solin, flax and canola. AB - Fatty acid profiles with emphasis on linoleic, linolenic, oleic, and conjugated linoleic acid (CLA) were compared in milk from dairy cows fed diets containing 3.25% supplemental fat and a control diet containing no supplemented fat. The fat was supplied by either whole ground solin, flax, or canola oilseed. Solin (linola) is a new cultivar of flax that contains 28% linoleic acid in the seed, Twelve multiparous Holstein cows were assigned to one of four dietary treatments. The experimental design was a 4 x 4 Latin square with each period consisting of 16 d for adjustment to the diet followed by a 5-d sampling period. Feed intake, milk yield, milk fat yield, and milk fat percentage were not affected by treatment. Adding solin, flax, or canola oilseed to lactation diets produced the highest proportions of linoleic (C18:2), linolenic (C18:3), and oleic (C18:1) acids, respectively, in the lipid fraction of the milk of the cows consuming these diets. The proportions of C6:0 to C16:1 were depressed in the milk fat of cows fed the oilseed diets, compared with the control diet. Increasing the lactation diet levels of C18:2, by using different oilseeds, increased CLA to 1.5% of milk fatty acids. The content of CLA in milk fatty acids, however, did not increase with the solin-supplemented diet compared with the canola supplemented diet even though the C18:2 level was higher in the former diet. PMID- 12086054 TI - Response of forage fiber degradation by ruminal microorganisms to branched-chain volatile fatty acids, amino acids, and dipeptides. AB - This study evaluated the effect of branched-chain volatile fatty acids (VFA; isobutyric acid, isovaleric acid), amino acids (valine, leucine), and dipeptides (valine-valine, leucine-leucine) on neutral detergent fiber (NDF) degradation by rumen microorganisms in vitro. The CP (%) and in situ NDF degradation rate (%/h) for alfalfa, bermudagrass, and pangolagrass hays, and napiergrass silage were 17.2 and 7.5, 4.7 and 3.1, 8.3 and 5.3, and 9.6 and 3.4, respectively. In vitro NDF digestibility was the lowest for bermudagrass; alfalfa and napiergrass were the highest. When the incubation contained more ammonia initially, digestibilities increased, but relative differences among forages were unchanged. Adding branched-chain VFA (2 mM) to incubations increased digestibilities more than controls on 15 out of 16 occasions. The effectiveness varied with isoacids and forages used. Amino acid (2 mM) or dipeptide (1 mM) addition consistently increased digestibility over controls. Amino acids further increased digestibility over corresponding isoacids on 14 occasions. Improvement in digestibility over control by leucine appeared to be greater than that by valine. Digestibilities with dipeptides were always greater than those with isoacids, except for one case. Dipeptide addition further increased digestibility significantly over corresponding amino acids on only six occasions, while percent improvement in digestibility numerically by dipeptides occurred in 10 cases. Valine-valine seemed to exert different effect than leucine-leucine, depending on initial ammonia availability. The results indicate that dipeptides could be more effective than isoacids and amino acids in improving NDF digestion. Forages with high CP content or rapid NDF degradation rate appeared to respond to additives to smaller degrees. PMID- 12086056 TI - Trans18:1 and 18:2 isomers in blood plasma and milk fat of grazing cows fed a grain supplement containing solvent-extracted or mechanically extracted soybean meal. AB - Thirty Holstein cows grazing mixed clover-grass pastures for 12 wk from May through July were fed a grain supplement containing solvent-extracted soybean meal (SES), or mechanically extracted soybean meal (MES) to determine whether differences in supplemental 18:2n6 fatty acid intake altered secretion of unsaturated fatty acids. Groups of 10 cows each were fed in two equal feedings a supplement (7.3 kg/d) containing ground corn plus either 1.8 kg of SES, 2.2 kg of MES, or 2.2 kg of MES plus 30 g of methionine hydroxy analog (Alimet; MESM). Fatty acid content (% of DM) of grass and clover in pastures averaged 1.9 and 1.5%, respectively. Concentration of 18:3n3 was higher in grass compared with clover (532 vs. 454 mg/g of total fatty acids). Yield of milk (32 kg/d average) and milk components did not differ by supplements. Total blood plasma fatty acids (mg/ml) during wk 4 were higher due to MESM (1.0) compared with MES (0.6) or SES (0.6). Cows fed MESM or MES had greater concentrations of 18:2n6, trans11-18:1, and cis9,trans11-18:2 in plasma compared with cows fed SES. The additional trans11-18:1 was found exclusively in plasma triglycerides, whereas the additional cis9,trans11-18:2 was found in plasma phospholipids and free fatty acids. Daily yields of 18:2n6, trans11-18:1 and cis9,trans11-18:2 in milk fat were greater for cows fed MES or MESM compared with SES. Results indicate yields of trans11-18:1, cis9, trans11-18:2, 18:2n6, and 18:3n3 in milk fat of pasture fed cows were enhanced by feeding a grain supplement containing mechanically extracted, rather than solvent-extracted, soybean meal. PMID- 12086057 TI - Effects of dietary protein solubility on nitrogen losses from lactating dairy cows and comparison with predictions from the Cornell Net Carbohydrate and Protein System. AB - This experiment determined the effects of dietary protein solubility on amount, form, and route of nitrogen loss in lactating Holstein dairy cows, and the ability of the Cornell Net Carbohydrate and Protein System (CNCPS) to accurately predict rumen microbial yield, serum urea N (SUN), milk urea N (MUN), and fecal N. Eighteen multiparous Holstein cows were assigned randomly to one of three dietary treatments that were similar in crude protein (17.7%) content but differed in their content of soluble intake protein (SIP). Dietary contents of SIP, as % of total CP were 30, 36, and 48%. The experimental period was 21 d, and total N balance collections were done during the last 5 d. As dietary content of SIP increased, excretion of urinary N increased quadratically, and it was the primary route of N excretion. Urinary excretion of purine derivatives (PD) responded quadratically as dietary SIP content increased. The CNCPS predicted a quadratic decrease in total metabolizable protein (MP) supply. No effect of dietary content of SIP was detected on MUN and SUN. The CNCPS predicted a quadratic decrease in SUN and MUN as dietary SIP increased. Results from this study indicated that changing the dietary content of SIP altered routes of N excretion in dairy cows, but had no effect on total N balance. The CNCPS did not adequately predict changes in SUN and MUN for cows fed diets varying in SIP. PMID- 12086058 TI - Dairy herd management practices that impact nitrogen utilization efficiency. AB - Improving the efficiency of feed N utilization by dairy cattle is the most effective means to reduce nutrient losses from dairy farms. The objectives of this study were to quantify the impact of different management strategies on the efficiency of feed N utilization for dairy farms in the Chesapeake Bay Drainage Basin. A confidential mail survey was completed in December 1998 by 454 dairy farmers in PA, MD, VA, WV, and DE. Nitrogen intake, urinary and fecal N, and efficiency of feed N utilization was estimated from survey data and milk analysis for each herd. Average efficiency of feed N utilization for milk production by lactating dairy cows (N in milk/N in feed x 100) was 28.4% (SD = 3.9). On average, farmers fed 6.6% more N than recommended by the National Research Council, resulting in a 16% increase in urinary N and a 2.7% increase in fecal N. Use of monthly milk yield and component testing, administration of bovine somatotropin (bST), and extending photoperiod with artificial light each increased efficiency of feed N utilization by 4.2 to 6.9%, while use of a complete feed decreased efficiency by 5.6%. Increased frequency of ration balancing and more frequent forage nutrient testing were associated with higher milk production, but not increased N utilization efficiency. Feeding protein closer to recommendations and increasing production per cow both contributed to improving efficiency of feed N utilization. PMID- 12086059 TI - Phosphorus partitioning during early lactation in dairy cows fed diets varying in phosphorus content. AB - The effect of dietary P content on P partitioning and excretion during early lactation was evaluated in 13 cows fed diets containing 0.34 (no supplementary P), 0.51, or 0.67% P. All cows were fed a common pre-partum total mixed ration (TMR) (0.28% P), followed by common TMR (0.51% P) for 7 d post-partum. On day 7, cows were randomly assigned to one of the three dietary P treatments. All treatment diets contained 16.6% CP, 15.2% ADF, 26.3% NDF, and 0.74% Ca. Total collections of milk, urine, and feces were conducted during weeks 3, 5, 7, 9, and 11 of lactation. Average body weight (591 kg), milk yield (47.9 kg/d), and DMI (25.2 kg/d) throughout the 10-wk trial were not affected by dietary P content. With increasing dietary P, however, linear increases in P intake (84.7, 135.2, and 161.5 g/d), fecal P (42.3, 87.5, and 108.6 g/d), urinary P (0.32, 1.28, and 3.90 g/d), and total P excretion (42.6, 88.8, and 112.5 g/d) were observed. Apparent P digestibility (49.0, 34.4, and 32.8% of P intake) decreased quadratically with increasing dietary P. Phosphorus balance was highly variable, but cows fed the 0.34% P diet were in negative P balance longer than were cows fed diets containing 0.51 or 0.67% P. With increasing dietary P, serum concentrations of inorganic P increased linearly, but serum Ca and Mg concentrations decreased. Increasing dietary P increased fecal and urinary P excretion in early lactation cows. Increased duration of negative P balance and changes in blood mineral concentrations suggest that cows fed low P diets mobilized more P from body reserves than cows on medium- and high-P diets. PMID- 12086060 TI - Lactoferrin supplementation to dairy calves. AB - Twenty-one Holstein calves (12 bulls, 9 heifers) were used to evaluate the effects of supplemental lactoferrin (0, 1, and 10 g/d) added to colostrum, milk, and milk replacer in a 56-d study. Calves fed lactoferrin (LF) weighed more during wk 2, 3, 4, 5, and 6 than control calves. Calves fed LF had increased preweaning daily weight gains and heart girth gains compared with calves not fed lactoferrin. Calves fed 1 g/d LF had a greater preweaning average daily gain than calves fed 10 g/d of lactoferrin. Hematocrit and serum Fe were unaffected by treatments. Calves were weaned when they had consumed 0.7 kg of calf starter grain for 2 consecutive d and were at least 28 d of age. All calves were weaned by 35 d of age, regardless of starter intake. Calves fed LF consumed more calf starter grain during the preweaning period and met weaning criteria at a younger age. This study indicates that supplementing calf diets with LF is advantageous to calf performance. PMID- 12086061 TI - Absorption of protein and IgG in calves fed a colostrum supplement or replacer. AB - Newborn Holstein bull calves (n = 32) were assigned to receive a colostrum supplement (CS) containing defibrinated bovine plasma or a colostrum replacer (CR) containing an immunoglobulin concentrate obtained by concentrating the immunoglobulin (Ig)G fraction of bovine plasma. The CS and CR contained 11.1 and 21.2% of dry matter as IgG, respectively. Each animal was fed two 454-g feedings at 1 and 8 h of age. The two feedings of CS and CR provided 95 and 187 g of IgG, respectively. Mean plasma IgG at 24 h of age was 8.0 and 13.6 g/L in calves fed CS and CR, respectively, indicating acceptable absorption of Ig from both sources. Mean apparent efficiency of IgG absorption in calves fed CS and CR were 33 and 30%, respectively, and did not differ between treatments. Mean plasma total protein at 24 h in calves fed CS and CR were 4.99 and 4.98 g/dl and did not differ between treatments. Increased plasma protein concentration from 0 to 24 h (4.5 g/L) was lower than the mean increase in plasma IgG concentration during the same period (10.3 g/L), indicating altered protein profile in the blood during the first 24 h of life. Correlation between plasma IgG and total protein at 24 h of age was significant within treatment, but the relationship between IgG and protein in plasma at 24 h varied between treatments. Predicted plasma total protein concentrations at 10 g of IgG/L of plasma at 24 h were 5.4 and 4.2 g/dl, in calves fed CS and CR, respectively. Prediction of plasma IgG concentration using total plasma protein may be inappropriate when calves are fed CS or CR. PMID- 12086062 TI - The use of bovine serum protein as an oral support therapy following coronavirus challenge in calves. AB - The objective of this experiment was to investigate the therapeutic efficacy of a supplemental bovine serum protein blend fed to calves challenged with virulent coronavirus. Twelve Holstein bull calves (approximately 3 wk of age) were allocated by initial body weight to Control (n = 5) and treated (n = 7) groups. On d 0, all calves were orally challenged with 1 x 10(7) plaque-forming units of virulent coronavirus isolate. Infection was allowed to progress for 24 h before treatment was started. On d 1, treated calves began receiving 160 g of dry bovine serum powder (16 g IgG) mixed into milk replacer powder (67 g) at both an a.m. and p.m. feeding. Control calves received only milk replacer powder (227 g) at both feedings. Response to coronavirus challenge and dietary treatment was monitored prior to a.m. and p.m. feeding by the collection of multiple clinical measures. Fecal consistency was decreased by coronavirus challenge but was not affected by dietary treatment. Mean daily rectal temperature and heart rate were not affected by dietary treatment. Average packed cell volume was higher in treated calves than in control (35.0 and 27.0%). Coronavirus challenge resulted in an immediate increase in respiration rate, decreasing by d 7. Control calves tended to have a greater average respiration rate compared with treated (28.7 vs. 26.8 breaths/min). Treated calves had a higher average feed intake than control (0.57 vs. 0.44 kg/d). These data suggest that bovine-serum supplemented milk replacer may decrease the severity of disease in young calves exposed to coronavirus. PMID- 12086063 TI - Prediction of ruminal pH from pasture-based diets. AB - This study identified suitable predictors of ruminal pH and identified relationships between ruminal pH and animal measures for diets based on fresh pasture. Animal and dietary variables (121 treatment means from six countries) were collated from 23 studies of lactating dairy cows fed pasture. Mean daily ruminal pH ranged from 5.6 to 6.7 across studies. Within studies, a low ruminal pH was associated with higher (P < 0.05; r2 > 0.40) microbial N flow from the rumen, total and individual volatile fatty acid concentrations, milk and milk component yields, and dry matter intake, and with lower (P < 0.05; r2 > 0.30) concentrations of milk fat, fat:protein, and acetate:propionate. Large variation between studies meant that these ruminal and production variables could not be used to make reliable predictions of ruminal pH in future pasture-based studies or feeding scenarios. Ruminal pH was positively related (P < 0.05; r2 < 0.15) to forage neutral detergent fiber (NDF) and NDF content within study, and negatively related (P = 0.001; r2 = 0.14) to nonstructural carbohydrate across studies. No single dietary variable, or group of variables, could be used to make a reliable prediction of ruminal pH. Estimates of effective fiber for diets containing only pasture were made using the Cornell Net Carbohydrate and Protein System ruminal pH equation. Mean effectiveness of fiber in pasture was 43% of NDF, and ranged from 17 to 78% across studies. High flows of microbial nitrogen, milk, milk fat yield, and dry matter intake suggested that the performance of cows fed high quality pasture was not limited when mean ruminal pH decreased to 5.8. PMID- 12086064 TI - Effects of feeding time on nitrogen capture by lactating dairy cows grazing rye pasture. AB - The objective of this experiment was to determine whether varying times at which a partial mixed ration was fed, either before or after grazing, affected N utilization from rye pasture and thus affected milk yield and components. Sixteen Holstein cows were fed a partial mixed ration (PMR) either at 0700, 0830, or 1100 h. Cows were milked at 0900 h and turned out to graze at 0930 h. Treatments represented feeding times 2.5 h and 1 h before grazing and immediately after grazing. The study was conducted as a 3 x 3 Latin square with three 17-d periods. There were no significant differences among treatments for pasture intake or yield of milk or milk components. Milk yield, fat %, and protein % were 29.4, 29.6, and 29.3 kg, 3.5, 3.5, and 3.4%, and 3.4, 3.5, and 3.4% for treatments, respectively. The milk urea levels were 15.6, 15.1, and 15.5 mg/dl, and were not different among treatments. Blood samples were collected on the last day of each period at 0645, 0845, 1045, 1200, and 1400 h. Blood urea nitrogen (BUN) was measured as an indicator of ruminal N capture. Concentrations were not significantly different among diets before grazing; however, they were significantly different among all treatments approximately 1 h after cows were removed from pasture. Cows fed at 0700 h, 2 h before grazing, maintained lower BUN levels across the 7 h during which the blood samples were collected. Cows that ate the PMR immediately after grazing maintained the highest BUN. Feeding a PMR to cows that graze at different times before and after grazing affected the capture of ruminal N, as indicated by differences in the levels of BUN, but there was no effect on yield of milk or milk components. PMID- 12086065 TI - Relationships among severity and duration of clinical mastitis and sire transmitting abilities for somatic cell score, udder type traits, productive life, and protein yield. AB - The objective of this study was to determine the relationships among severity and duration of clinical mastitis during first and second lactation and sire transmitting abilities for somatic cell score, udder type traits, productive life, and protein yield. Recording of clinical episodes began at first parturition for 1704 Holstein cows (in six Pennsylvania herds and one Nebraska herd) and continued into second lactation for 1055 of these cows. A total of 456 cows (sired by 168 bulls) had at least one clinical episode during first lactation, and 230 cows (sired by 100 bulls) had at least one clinical episode during second lactation. A severity code from 1 (normal milk) to 5 (acute systemic mastitis) was assigned daily (for up to 30 d after detection) to all quarters that had clinical mastitis. Only the severity codes for the first clinical episode to occur during first and second lactation are considered here. The initial and maximum severity codes, as well as the natural logarithms of both the sum of severity codes that were above normal (> 1) and the total days severity codes were above normal were regressed on herd (a classification variable), age at first calving, days in milk at clinical detection, and sire transmitting abilities taken one at a time. Linear and nonlinear effects were estimated for sire transmitting abilities. Separate analyses were conducted on dependent variables that considered severity and duration of clinical mastitis from: all organisms, coagulase-negative staphylococci, coliform species, streptococci other than Streptococcus agalactiae, and the most common environmental organisms (coliform species and streptococci other than Streptococcus agalactiae). Daughters of sires that transmit the lowest somatic cell score had the least severe and shortest clinical episodes from environmental organisms during first lactation. Selection for lower somatic cell score may reduce the severity and duration of clinical episodes from environmental organisms during first lactation. PMID- 12086066 TI - Estimation of quantitative trait loci effects in dairy cattle populations. AB - Standard animal model programs can be modified to include the effect of a quantitative gene, even if only a fraction of the population is genotyped. Five methods to estimate the effect of a diallelic quantitative gene affecting a quantitative trait were compared to a standard animal model (model I) on simulated populations, based on mean squared errors and bias. In models II, III, and IV complete linkage between a single genetic marker and the quantitative trait gene was assumed. In models II and III the elements of the incidence matrix for the gene effect were 0 or 1 for genotyped individuals, and the probabilities of the possible candidate gene genotypes for individuals that were not genotyped. In model III segregation analysis was used to compute these probabilities. If only some of the cows were genotyped, the model III estimates were nearly unbiased, while model II underestimated the simulated effects. When only sires were genotyped, model II overestimated the simulated effect. In models V and VI two markers bracketing the quantitative gene with recombination frequencies of 0.1 and 0.2 with the quantitative gene were simulated, and the algorithm of Whittaker et al. (1996) was used to derive estimates of gene effect and location. In model V marker allele effects were included in the animal model analysis. In model VI, the model I genetic evaluations were analyzed. Model V estimates for both effect and location of the quantitative gene were unbiased, while model VI estimates were only 0.25 of the simulated effect. PMID- 12086067 TI - Effects of age and calving season on lactation curves of milk production traits in Italian water buffaloes. AB - Test day (TD) records of milk production traits (milk yield, fat, and protein percentages) of 534 Italian buffalo cows were analyzed with a mixed linear model in order to estimate lactation curves pertaining to different ages at calving and different seasons of calving. Milk yield lactation curves of younger animals were lower than those of older animals until 20 wk from parturition. No effect of age at calving could be observed for fat and protein percentages. Season of calving affected milk yield only in the first phase of lactation, with the lowest production levels for summer calvings; no effect could be observed on fat and protein contents. Average correlations among TD measures within lactation were 0.59, 0.31, and 0.36 for milk yield, fat, and protein percentages, respectively. Five standard linear functions of time were able to reconstruct the average lactation curves. Goodness of fit was satisfactory for all models considered, although only the five-parameter model was flexible enough to fit all the three traits considered with excellent results. PMID- 12086068 TI - Indirect prediction of herd life in Guernsey dairy cattle. AB - Production and type data were used to investigate the relationships of these traits with herd life data in US Guernsey cows that calved from 1985 through 1990. Two definitions of herd life were used: actual days from birth to disposal (true herd life) and herd life adjusted for milk production (functional herd life). Genetic parameters were calculated with data from cows that had an opportunity to reach 84 mo of age (n = 18,725). Linear type traits were preadjusted for stage of lactation and age at classification. True herd life was preadjusted for age at first calving and for functional herd life, within herd year quartile ranking for milk yield. The (co)variance components for true and functional herd life, milk, fat, protein, and 15 linear type traits were estimated with multiple-trait REML in an animal model. Heritability estimates for true and functional herd life were 0.12 for both traits. Estimated genetic correlations of herd life with body size traits were from -0.14 to -0.29, with feet and leg traits were from -0.10 to 0.06, and with udder traits were from 0.09 to 0.24. These correlation parameters were used for indirect prediction of herd life from available production and type information in Guernseys. PMID- 12086069 TI - The effect of pathogen-specific clinical mastitis on the lactation curve for somatic cell count. AB - Data from 274 Dutch herds recording clinical mastitis (CM) over an 18-mo period were used to investigate the effect of pathogen-specific CM on the lactation curve for somatic cell count (SCC). Analyzed pathogens were Staphylococcus aureus, coagulase-negative staphylococci, Escherichia coli, Streptococcus dysgalactiae, Streptococcus uberis, other streptococci, and the culture-negative samples. The dataset contained 178,754 test-day records on SCC, recorded in 26,411 lactations of 21,525 cows of different parities. In lactations without both clinical and subclinical mastitis, SCC was high shortly after parturition, decreased to a minimum at 50 days in milk (DIM), and increased slowly toward the end of the lactation. Effects of CM on lactation curves for SCC differed among the pathogens isolated. Before a case of clinical E. coli mastitis occurred, SCC was close to the SCC of lactations without both clinical and subclinical mastitis, and after the case of CM had occurred, SCC returned rather quickly to a low level again. Similar curves were found for lactations with cases of CM associated with culture-negative samples. Before a case of clinical Staph. aureus mastitis occurred, average SCC was already high, and it remained high after the occurrence. Effects of CM associated with Strep. dysgalactiae, Strep. uberis, and other streptococci on the lactation curve for SCC were comparable. They showed a continuous increase in SCC until the case of pathogen-specific CM occurred, and afterwards SCC stayed at a higher level. Using SCC test-day records, these typical characteristics of each pathogen may be used to find more effective indicators of CM. PMID- 12086070 TI - Analysis of age-specific predicted transmitting abilities for final scores in Holsteins with a random regression model. AB - Conformation final scores obtained by the Holstein Association were used in this study. Five subsets were sampled from the original complete data, which contained about 7 million records, and (co)variance components for additive genetic, permanent environmental, and residual effects were estimated with a multiple trait (MT) model at different ages of cows and with a random regression model. Transmitting abilities were predicted with the random-regression (RR) model and with the repeatability model for the national evaluation, using the entire data set. The RR model included fixed and random regressions on age at classification for additive genetic and permanent environmental effects. Additive genetic and residual variances estimated with the MT model increased with aging. Genetic correlations between age groups decreased with their distance. Heritability and repeatability estimates with the RR model were 0.27 and 0.88 on average, increasing with cow's age. Correlations between transmitting abilities predicted with the RR model at various ages and with the repeatability model were in the range of 0.89 to 0.99. In the 1980s, predicted transmitting abilities (PTA) of sires increased with their daughters' age, but in 1991 and 1995, the PTA decreased. In general, sires whose daughters were evaluated at mature ages tended to have higher PTA. For sires and cows, genetic trends estimated from the repeatability model and from the RR model were similar. The low genetic trend of cows at 60 mo for recent years was due to many grade cows that had only single records and little pedigree information. PMID- 12086071 TI - Productivity of grazing Holstein cows in Atlantic Canada. AB - The feasibility and profitability of management-intensive grazing (MIG) in Atlantic Canada was studied. Productivity of MIG plus concentrate (1 kg:3 kg of milk) was compared with that of confinement feeding (Confined) using haylage (1996) or corn silage (1997) as 50% of dry matter (DM) in a total mixed ration (TMR). Each year, two groups of 10 Holsteins were used. In 1996, pasture CP content increased from 15.2 to 18.0% of DM, while those of acid detergent fiber (ADF) and neutral detergent fiber (NDF) decreased (33.6 to 23.8%, and 55.6 to 35.4%, respectively) between July 10 and August 28. In 1997, pasture crude protein (CP) content decreased from 30.0 to 15.7%, while ADF and NDF increased (24.9 to 35.8% and 53.5 to 67.4%, respectively) from June 16 to July 14. Because pasture biomass was reduced by winter-kill followed by drought in 1997, MIG cows were supplemented with TMR. Estimated pasture DM intakes ranged from 14.2 to 18.1 kg/d per cow. Milk yields averaged 29.5 and 30.4 +/- 0.43 kg/d in 1996 and 32.5 and 31.8 +/- 0.61 kg/d in 1997 for Confined and MIG cows, respectively. In 1996, the dietary treatment x time interaction was significant. Cows in MIG had higher yields than those in Confined later in the trial, while fat and CP concentrations were usually lower in milk from MIG cows. Lower body weights resulted with MIG. Few differences between production systems were significant in 1997. Milk revenue (Canadian $/d) from Confined cows was higher (14.03 vs. 13.77 in 1996 and 16.10 vs. 15.39 in 1997), but partial profitability of the MIG system was marginally greater in both years. PMID- 12086072 TI - Extent and benefits of multi-country progeny testing of young dairy sires. AB - One of the current trends within the artificial insemination industry is to progeny test young dairy bulls in multiple countries. The objectives of this study were to assess the extent of multi-country progeny testing and to measure the corresponding gains in reliability of international breeding value estimates. Data of Holstein bulls that were born between July 1, 1992, and December 31, 1994, and progeny tested in countries that participate in the International Bull Evaluation Service were used in the present study, because these were the youngest bulls that had completed multi-country progeny testing before the study. Based on August 1999 international sire evaluation data, a total of 562 bulls from 10 countries were multi-country sampled for production traits during this 2.5-yr period, and 233 bulls from seven countries were multi-country sampled for type traits. The United States, Canada, The Netherlands, France, and Germany were most active in multicountry progeny testing, and Germany, New Zealand, Australia, France, and The Netherlands were the most common countries of foreign sampling. Mean reliabilities of international breeding values were calculated within each country. Means for milk yield were 0.89 for single-country sampled bulls with local progeny (i.e., progeny in the home country), 0.71 for single-country sampled bulls with no local progeny, 0.90 for multicountry sampled bulls with local progeny, and 0.78 for multi-country sampled bulls with no local progeny. Mean reliabilities for teat placement for these groups of bulls were 0.80, 0.71, 0.88, and 0.83, respectively, and means for rear udder width were 0.79, 0.60, 0.85, and 0.68, respectively. Gains in reliability in the country of foreign sampling were greatest when foreign progeny were located in countries that had low genetic correlations with the home country. PMID- 12086073 TI - Persistence of anti-leptospiral IgM, IgG and agglutinating antibodies in patients presenting with acute febrile illness in Barbados 1979-1989. AB - The persistence of anti-leptospiral IgM and IgG antibodies and agglutinating antibodies was studied in serologically confirmed cases of severe leptospirosis during the acute illness and over periods of several years after recovery. The antibody response in non-leptospirosis patients presenting to hospital with similar symptoms over the same period of time was used to estimate the background antibody level to leptospirosis in the community. All patients enrolled in the study had blood samples collected twice in the acute stage of illness, once during convalescence and then annually from the time of initial hospitalisation until the end of the study period. Six hundred and thirty-eight patients presented to hospital with acute febrile illness, of whom 321 were diagnosed with leptospirosis. Patients who had severe leptospirosis commonly remained seropositive, with IgM, IgG and agglutinating antibodies detectable for several years after infection. A significant proportion of cases had high titres of agglutinating antibody detectable by the microscopic agglutination test (> or = 800). There were marked differences in the magnitude and duration of persistence of agglutinating antibodies directed against different serogroups. More than 20% of cases with evidence of infection with serogroup Autumnalis retained titres of >800, 4 years after the acute illness. In one case a titre of 800 was detected 11 years after infection. Persistence of agglutinating antibody titres can create problems in interpretation of serological results and make it impossible to estimate the time of infection, given a specific titre. This study demonstrates that in endemic areas where seroprevalence is high, use of a single elevated titre is not reliable to define a current infection. PMID- 12086074 TI - Family history and the risk of coronary heart disease: comparing predictive models. AB - Family history is commonly used when evaluating coronary heart disease (CHD) risk yet it is usually treated as a simple binary variable according to the occurrence or non-occurrence of disease. This definition however fails to consider the potential components of a family history which may in fact exert different degrees of influence on the overall risk profile. The purpose of this paper is to compare different predictive models for CHD which incorporate family history as either a binary variable or different types of family risk indices in terms of their predictive ability. Models for estimating CHD risk were constructed based on usual risk factors and different family history variables. This construction was accomplished using logistic regression and RECursive Partition and AMalgamation (RECPAM) trees. Our analyses demonstrate the importance of using more sophisticated definitions of family history variables compared to a simple binary approach since this leads to a significant improvement in the predictive ability of CHD risk models. PMID- 12086075 TI - Mortality and temperature in Oslo, Norway, 1990-1995. AB - The associations between temperature and daily mortality was studied among the citizens of Oslo, Norway, 1990-1995. Data on daily mortality were linked with daily temperatures, relative humidity, wind velocity and air pollution. At temperatures below 10 degrees C, a 1 degrees C fall in the last 7 days average temperature increased the daily mortality from all diseases by 1.4%, respiratory diseases 2.1%, and cardiovascular diseases 1.7%. Above 10 degrees C, there was no statistically significant increase in daily mortality, except for respiratory mortality, which increased by 4.7% per 1 degrees C increase in the last 7 days average temperature. Daily mortality in Oslo increases with temperatures falling below 10 degrees C. The increase starts at lower temperatures than shown in warmer regions of the world, but at higher temperatures than in regions with even colder climates. As well insulated and heated dwellings are standard in Norway today, more adequate clothing during outdoor visits is probably the most important preventive measure for temperature related mortality. PMID- 12086076 TI - Joint effects of alcohol consumption and cigarette smoking on atherogenic lipid and lipoprotein profiles: results from a study of Chinese male population in Taiwan. AB - BACKGROUND: The present study examines the effect of joint exposure to cigarette smoking and alcohol intake on serum levels of total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL C), and triglyceride (TG) among Chinese male adults in Taiwan. METHODS: A sample of 3311 men aged 20-59 years who reported having stable smoking and drinking behaviors during the period between January 1995 and December 1996 was selected from a periodic health checkup population. Serum lipids and lipoprotein cholesterol fractions were measured on fasting blood samples taken from participants. Statistical methods of analysis of variance and analysis of covariance were conducted to examine the associations of different smoking drinking behavioral patterns with lipid and lipoprotein levels. RESULTS: In the observed population, the percentages of men who had stable cigarette smoking and alcohol consumption behaviors were 39.5% (1,307/3,311) and 27.0% (895/3,311), respectively. Mean values of TC and TG increased significantly and monotonically with increasing levels of cigarette smoking and alcohol consumption. In addition, alcohol intake was significantly associated with increased HDL-C and reduced LDL C levels in a dose-dependent manner. More interestingly, the effect of alcohol consumption on LDL-C (negative) and TG (positive) levels was substantially greater for heavy smoker (>20 cigarettes/day) than for light smokers (< or = 20 cigarettes/day) and non-smokers, while alcohol intake exerted a strong positive influence on HDL-C concentration regardless of levels of cigarette smoking. CONCLUSIONS: In this Chinese male population, cigarette smoking and alcohol consumption were confirmed to have similar effects on lipid and lipoprotein levels as in Caucasians. More interestingly, a significance of joint exposure to smoking and drinking in predicting lipid and lipoprotein levels was evident. These data indicate the importance of multifactorial interventions to obtain more favorable lipid and lipoprotein levels in the population. PMID- 12086077 TI - Changes of the epidemiology and the clinical picture of tularemia in Southern Moravia (the Czech Republic) during the period 1936-1999. AB - The author has processed data on early occurrence of tularemia in Southern Moravia collected from literature, materials of epidemical ward of the Regional Hygienic Station (RHS), Brno, and clinical documentation of patients treated for tularemia at the Clinic of infectious diseases, Brno. Tularemia has been appearing in Southern Moravia since autumn 1936. At that time, 290 people contracted the external (mainly ulceroglandular) form of this disease by handling tularemic hares. During the 60s, large severe epidemics of professionally acquired pulmonary forms of tularemia, mainly in workers in 'cold divisions' of sugar factories, were occurring. The 25-year period of low occurrence of disease, which followed, was interrupted in 1978 with the curious epidemic of tularemia contracted through the alimentary tract in an old people's home. The transmission medium of the infectious agent was juice which had been made from fallen apples infected by little mouse rodents. In autumn 1994, the number of tularemia cases rose again and, during the season of 1998-1999, 115 cases of tularemia contracted mainly by handling tularemic hares were recorded. The author demonstrated the relationship among sources and modes of transmission of Francisella tularensis and clinical form of the disease using the group of patients treated at the Clinic of infectious diseases in Brno. PMID- 12086078 TI - Familial history, age and smoking are important risk factors for disc degeneration disease in Arabic pedigrees. AB - The present study used computed tomography imaging to evaluate the extent and pattern of the intergenerational transmission of spinal disc degeneration disease (DDD) in complex pedigrees. Contribution of a number of the potential covariates was also studied using univariate and multivariate logistic regression analysis, as well as two types of complex segregation analysis models. Among 161 individuals studied, DDD was diagnosed in 60 individuals. The number of protruded discs varied from 1 to 4, mostly in lumbar or lumbosacral regions. The average age at onset of the disease was similar for both women (36.0 years) and men (34.8 years). The proportion of the individuals affected by the DDD status of their parents ranged from 10% in families of two healthy parents to 55.5% of two affected parents (p < 0.01). The results of the logistic regression analyses and complex segregation analysis were qualitatively the same: DDD status of parents, age and smoking were the main risk factors for disc herniation in the Arabic families we examined. All analyses showed a predominating role of the family history as a risk factor for DDD in offsprings. It showed, for example, four times higher risk at age 50 for individuals with two affected parents vs. those who have two non-affected parents. However, the results of models-fitting genetic analysis, did not confirm a monogenic Mendelian pattern of inheritance. PMID- 12086079 TI - Epidemiological assessment of the influence of socio-family factors in adolescent pregnancy. AB - Pregnancy in adolescence constitutes a serious public health problem at these ages. It de-stabilizes family homeostasis; family and affective support received by the adolescent is recognized to be a factor of influence in the problem. The objective of the research presented is the empirical study in Cadiz, SW Spain, of the socio-family support received by a group of adolescent mothers, and the influence of this on the development of the pregnancy and the infant, in comparison with a group of adult mothers. A descriptive study of 305 adolescent mothers and 285 adult mothers who gave birth in 1996 and attended the family planning centers of the Primary Health Care District of Cadiz, Spain, in 1997. The variables used included socio-demographic data of the adolescents, information on the pregnancy and birth, on the health of the baby, and on social and family support (by means of the Duke-Inc and Apgar family questionnaires). In the statistical study of the data, frequencies, statistical association, and disequalities of prevalence in the variables were determined. Of the totals, 81.6% (77-86) of the adolescent mothers were unmarried, compared with 22% (17-26) of the adult mothers; 82.3% (78-87) of the adolescents were living with their parents, compared with 39.3% (34-45) of the adults. Family support received is greater among the adolescents (p < 0.01). In this group, in line with increasing family support, there is improvement in--the decision to continue the pregnancy (p < 0.0001), frequency of check-ups (p < 0.0001), less problems in the pregnancy (p < 0.01), differences in the anthropometric values of the baby at birth (p < 0.0001), and breast-feeding (p < 0.001). The conclusion is reached that juvenile pregnancy is not synonymous with more risky pregnancy. There is a positive correlation between the socio-family support received and the general success of the pregnancy and better health in the mother and baby. PMID- 12086080 TI - Tetanus immunization in pregnant women. Serum levels of antitetanus antibodies at time of delivery. AB - The objective of this study was to determine the tetanus immunity status of pregnant women at the time of delivery according to tetanus toxoid (TT) vaccination coverage during their most pregnancy. The serum anti-tetanus antibody levels of 493 mothers who had live births at a hospital in Ankara were determined by enzyme-linked immunosorbent assay (ELISA). Protected women were defined as those with serum antibody levels > or = 0.6 IU/ml. Overall, 69.0% of mothers had protective tetanus antibody titers at the time of delivery. The rates of protection for mothers who had received no vaccination, one TT dose, or two TT doses during pregnancy were 46.4, 93.5, and 95.6%, respectively. Vaccinating every pregnant woman with at least one dose of TT would be an affordable and effective way to protect against neonatal tetanus, and would be a step toward eliminating the deaths that continue to occur due to this preventable disease in Turkey. PMID- 12086082 TI - Reduction in fertility in male greenhouse workers exposed to pesticides. AB - The paper examines the possible interference of pesticide exposure on male fertility, by studying the time to pregnancy (TTP) in the first pregnancy of 127 greenhouse workers and 173 controls. The TTP of exposed and control population, analysed by logistic regression model, has shown an increase in the risk of conception delay among the greenhouse workers with high exposure (OR:2.4; 95% CI: 1.2-5.1). PMID- 12086081 TI - The ecological relationship of tobacco smoking to the prevalence of symptoms of asthma and other atopic diseases in children: the International Study of Asthma and Allergies in Childhood (ISAAC). AB - This study examined the relationship between parental smoking and asthma and other atopic diseases at the ecological level. The prevalence of atopic symptoms in 6-7- and 13-14-year old children was assessed in 91 centres (from 38 countries) and 155 centres (from 56 countries) respectively in the International Study of Asthma and Allergy in Childhood (ISAAC). These symptoms were related to prevalence of tobacco smoking for each country by gender as reported by the World Health Organisation. There was a significant negative association between the prevalence of smoking by men and the prevalence of symptoms of asthma and rhinitis, but not eczema in the 13-14-year age group. There was a significant positive relationship between prevalence of smoking by women and prevalence of wheeze in the last 12 months, but not for any other symptoms in the 13-14-year age group. In the 6-7-year age group, there was a significant negative correlation between prevalence of smoking by men and the prevalence of wheeze causing sleep disturbance and a close to significant negative association at the 5% level with the prevalence of wheeze in the last 12 months and rhinitis. Thus, for the countries that are included in this analysis, countries that have high adult male smoking rates have a lower risk of asthma and rhinitis symptoms in children. It should be stressed that this analysis does not involve information on individual exposures and therefore does not contradict the well-established association of active and passive smoking in individuals with the occurrence of asthma symptoms in the same individuals. Rather, it indicates that this well established individual-level association does not account for the international differences in asthma prevalence, and that other risk factors for asthma must be responsible for the observed international patterns. PMID- 12086083 TI - Review of the utilisation of a university hospital in Barcelona (Spain): evolution 1992-1996. AB - BACKGROUND: To test the applicability of the appropriateness evaluation protocol (AEP) as a tool for reviewing hospital utilisation. To quantify and to compare the rate of inappropriate admissions and amount of in-hospital days, emphasising the main causes and factors in the hospital associated with inappropriateness during the studied periods of time. PATIENTS AND METHODS: Two retrospective studies were carried out, the first one in 1992, when 2048 clinical histories were analysed, and the second in 1996, with 1099 reviewed histories. The tool used for the evaluation of the level of hospital utilisation is the AEP. RESULTS: The proportion of admissions considered to be inappropriate was 25% (95% CI: 20.8 24.5) in 1992, and 16% (95% CI: 13.8-18.2) in 1996. Premature admission was the most frequent cause of inappropriateness in both periods. The logistic regression model built for the dependent variable admission showed the following variables to be associated to inappropriateness: scheduled admission (OR: 15; 95% CI: 10.8 20.7) and (OR: 10; 95% CI: 6-16.5), weekend admission (OR: 2; 95% CI: 1.3-1.2) and (OR: 2; 95% CI: 1.2-2.3), for 1992 and 1996, respectively. The rate of inappropriate in-hospital stays in 1992 was 29% (95% CI: 28.3-29.6), and 13.5% (95% CI: 12.7-14.3) in 1996. Hospital organisational problems were the main cause of inappropriate in-hospital days in 1992, and diagnostic/ therapeutical tests that could be performed ambulatorily ranked first in 1996. The logistic regression model built for the dependent variable in-hospital days had the following independent variables associated to its inappropriateness: discharge on weekdays (OR: 1.4; 95% CI: 1.2-1.8) and (OR: 0.6; 95% CI: 0.5-0.8) and length of stay (OR: 2; 95% CI: 1.8-2.4) and (OR: 2.4; 95% CI: 2.3-2.5), for 1992 and 1996 respectively, among other variables. CONCLUSION: Periodic checking of the utilisation levels with the application of methods such as the AEP will contribute towards adapting hospital management in the more competitive current setting. PMID- 12086084 TI - Impact of selected geographical and clinical conditions on thrombolysis rate in myocardial infarction in three departments of France. AB - Our objective was to assess the impact of selected geographic factors and patients' characteristics on thrombolysis rates in patients resident and hospitalised for acute myocardial infarction in three departments of the Rhjne Alpes Region (France). We used a two-level hierarchical model to estimate and explain geographic areas' specific effects. Old subjects and women were less frequently treated than young subjects and men. Severe, non-anterior and non-Q wave myocardial infarctions were associated with lower thrombolysis rates. It was also lower in patients with a pulmonary chronic disease, a cancer, a peripheral arterial disease, a history of cerebrovascular accident or transient ischaemia attack, and in patients with a psychiatric disorder. After adjusting for patients' characteristics, significant variations in thrombolysis rates remained between geographic areas (up to 3.2 times). These variations seem to be partly explained by distance or isolation: a longer distance to the closest hospital or a high degree of isolation seem to lower the probability of thrombolytic therapy. Several other sources of treatment variation between the studied geographic areas remain unexplored. These factors, especially those that augment the delay to treatment, are to be identified in order to augment fibrinolysis usage and reduce inter-area heterogeneity. PMID- 12086085 TI - Prevalence of hepatitis B, hepatitis C, syphilis and HIV in Georgian blood donors. AB - Prevalence of human immunodeficiency (HIV), hepatitis B (HBV), hepatitis C (HCV) virus and syphilis in the population of blood donors in Georgia has been investigated. Out of 4970 donors 7.3% had anti-HCV (6.9% confirmed), HbsAg was positive in 4.1% (3.4% confirmed), Seroprevalence of Syphilis was 2.3%. Three individuals had HIV. Prevalence of HCV and HBV in Georgia is higher than national prevalence estimates of viral hepatitis in neighboring countries. PMID- 12086086 TI - Does oral polio vaccine cause intussusception in infants? Evidence from a sequence of three self-controlled cases series studies in the United Kingdom. AB - BACKGROUND: The use of live oral poliomyelitis vaccine (OPV) has led to the elimination of poliomyelitis disease in many countries since licensure in 1960. The discovery of an increased risk of an intestinal obstruction known as intussusception following live rotavirus vaccination raised questions about the possibility of a link between live OPV and intussusception. METHODS: Three self controlled case-series studies were carried out. The first was exploratory and included 218 intussusception episodes from hospital admissions data linked to vaccination records in the Thames region. The two subsequent studies, which used further hospital admissions data and the General Practice Research Database (GPRD) included 107 and 198 episodes respectively and were used to test hypotheses generated in the first study. RESULTS: In the exploratory study risk periods of up to 6 weeks after each dose were examined. The only period with some evidence of an increased risk was the 14-27-day period after the third dose (relative incidence (RI) = 1.97, p = 0.011). The second hospital admissions study and the GPRD study showed no evidence of an increased relative incidence in any putative risk period and did not confirm the increased risk in the 14-27-day period after dose 3 with a combined RI of 1.03. CONCLUSIONS: The sequence of studies does not support the hypothesis that OPV causes intussusception. The increased RI in the first study may be explained as a chance finding due to the number of risk periods examined and highlights the need for caution when looking at many risk periods without an a priori hypothesis. PMID- 12086087 TI - ApoE polymorphism in a small Mediterranean island: relationships with plasma lipids, lipoproteins and LDL particle size. AB - Polymorphisms of apoE gene are able to modulate lipoprotein metabolism at different steps and to influence LDL-cholesterol (LDL-C) levels and also other lipoproteins features. Population studies documented large differences in the frequency of apoE alleles which could be even related to the prevalence of cardiovascular disease. In this study we evaluated the apoE genotypes and allele frequency in 576 subjects living in a small island in the Tyrrhenian Sea and the relative contribution of apoE polymorphism on plasma lipid and lipoprotein profile, including LDL particle size. We found a cumulative frequency of 0.073, 0.866 and 0.061 for epsilon2, epsilon3 and epsilon4 alleles respectively. Moreover epsilon3 subjects had only triglyceride levels significantly lower and LDL-C and lipoprotein (a) (Lp(a)) levels higher than epsilon2 carriers. LDL particle size was significant smaller in epsilon2 subjects than both epsilon3 and epsilon4 carriers, but the difference disappeared when data were adjusted for triglycerides. In conclusion we have provided further evidence of a low prevalence of epsilon4 allele in a Mediterranean population which may represent a genetic protective factor of these populations. Environmental factors, such as diet, occurring in this area may have attenuated the influence of this gene on plasma lipoproteins. PMID- 12086088 TI - Neonatal late-onset bloodstream infection: attributable mortality, excess of length of stay and risk factors. AB - BACKGROUND: Bloodstream infection represents a major threat among neonates under intensive care with considerable impact on morbidity and mortality. This study evaluated extra stay, attributable mortality and the risk factors associated with late-onset bloodstream infection (LO-BSI) among neonates admitted to a neonatal intensive care unit during a 4-year period. METHODS: A retrospective matched cohort study was conducted. For each case, there was one control patient without LO-BSI matched for sex, birth weight, gestational age, duration of hospitalization prior to the date of LO-BSI in the respective cases, underlying illness and birth date. A novel test, sequential plan, was employed for attributable mortality analysis in addition to standard tests. Multiple logistic regression was employed for risk factor analysis. RESULTS: Fifty pairs of cases and controls were compared. LO-BSI prolonged hospital stay of 25.1 days in pairs where both subjects survived. Overall attributable mortality was 24% (95% CI: 9 39% p < 0.01) and specific attributable mortality due to Staphylococcus epidermidis was 26.7% (95% CI: 23-30.4%; p = 0.01). Blood and/or blood components transfusion was independently associated with neonatal LO-BSI (OR: 21.2; 95% CI: 1.1-423). CONCLUSIONS: LO-BSI infection prolongs hospital stay and is associated with increased mortality among neonates. In the present series, blood transfusion was a significant risk factor for LO-BSI. PMID- 12086089 TI - Prevalence, risk factors and evaluation of a screening strategy for chronic hepatitis C and B virus infections in healthy company employees. AB - A cross-sectional study was carried out in employees of 17 Greek companies with the aim of assessing the prevalence of hepatitis B (HBV) and hepatitis C (HCV) virus, identifying associated prognostic/risk factors and evaluating the effectiveness of a questionnaire as a pre-screening tool. All participants were asked to complete a questionnaire and a random sample of them was asked to provide a blood sample for hepatitis B core antibody (anti-HBc), hepatitis B surface antigen (HBsAg) and antibodies to hepatitis C (anti-HCV) testing. Individual questions or combinations of them were evaluated in terms of their ability to detect HBV or HCV(+) cases. Of 9085 eligible employees, 6074 (67%) completed the questionnaire. Of 990 samples obtained, 19.9% were anti-HBc(+), 2.6% HBsAg(+) and 0.5% anti-HCV(+). All anti-HCV(+) cases had multiple parenteral risk factors. Multiple logistic regression identified associations between anti HBc and older age, family members with chronic hepatitis, job category and history of transfusion before 1992. HBsAg(+) was associated with older age and history of transfusion before 1992. None of the risk/prognostic factors had sufficient sensitivity and specificity for HBV but report of at least one risk factor identified all HCV(+) cases. Anti-HCV screening of those with at least two parenteral risk factors not only identified all anti-HCV(+) cases but also resulted in 86% decrease in the screening cost. Under the light of recent treatment advances, targeted questionnaire-based screening of asymptomatic people may prove to be a cost-effective way to face hepatitis C. PMID- 12086090 TI - Phenotypic characterization of Trichophyton rubrum isolates from two geographic locations in Brazil. AB - To characterize possible Trichophyton rubrum phenotypes, which circulate in two Brazilian localities, we tested 53 isolates of this dermatophyte for their ability to assimilate several carbon sources, for keratinase, proteinase, phospholipase, lipase and desoxiribonuclease (DNase) secretions, and for their susceptibility to the antifungals fluconazole, ketoconazole and itraconazole. For each method, the isolates were submitted to similarity analysis and the methods were evaluated for their discriminatory indexes. None of the isolates were capable of assimilating arabinose, dulcitol, lactose, melibiose, ribose and xylose, while all of the isolates assimilated maltose, sucrose and sorbitol. However, adonitol, cellobiose, dextrin, erythritol, fructose, galactose, inulin, mannitol, mannose, raffinose, rhamnose and trehalose were assimilated by some isolates but not by others. All isolates secreted keratinase and DNase, while none secreted phospholipase. Proteinase and lipase were secreted only by some isolates. All but four isolates were resistant to fluconazole, most of them were sensitive to ketoconazole and all were sensitive to itraconazole. Carbohydrate assimilation was the method that presented the highest discriminatory index, and also the method that displayed the largest number of biotypes. Taken together, these data suggest that significant phenotypic variations exist among T. rubrum isolates. They seem to occur independently from their geographic origins. PMID- 12086091 TI - Incidence of pulmonary tuberculosis and HIV coinfection in the province of Seville, Spain, 1998. AB - Using the National Notifiable Disease Surveillance System (NNDSS) and Minimum Basic Data Set (MBDS) for hospital discharges we evaluated pulmonary tuberculosis (PTB) surveillance and estimated real PTB incidence and human immunodeficiency virus (HIV) coinfection in Seville in 1998. In addition, we assessed the sensitivity and the positive predictive value (PPV) of the surveillance system. Real incidence of pulmonary tuberculosis was estimated by the capture-recapture method. In 1998, the province of Seville reported 225 cases of pulmonary tuberculosis to the NNDSS, an incidence of 13.2 per 100,000 population. Of the 225 cases reported, 18.2% presented with HIV coinfection, while a total of 194 had confirmed diagnoses. The MBDS accounted for 106 new cases. Of these, 24.8% presented with HIV coinfection and were 58% less likely to be reported to the NNDSS (25-76%, p = 0.001). Applying the capture-recapture method, 426 cases were estimated, an overall incidence of 25.6 per 100,000 population (21.5-28.8). Completeness for each source was similar (47%), and for both jointly was 72.7%. The NNDSS had a sensitivity of 65.3% and a PPV of 89.3%. In conclusion, the NNDSS underestimates PTB and PTB-HIV coinfection in Seville. The high incidence observed in young adults suggests a high degree of tuberculosis endemicity. Hospital records provide a readily accessible, low-cost means of estimating disease incidence. PMID- 12086092 TI - Diagnosis and treatment of type 2 diabetes in three Belgian regions. Registration via a network of sentinel general practices. AB - BACKGROUND: The objective of this descriptive study is to investigate the incidence of type 2 diabetes in sentinel general practices in three regions in Belgium and to describe the patient characteristics and the start of the medical management after diagnosis. METHODS: For two successive years all patients who were newly diagnosed with type 2 diabetes were registered. Two weeks after inclusion, the registering physician completed an extensive questionnaire. This questionnaire dealt with risk factors, biochemical parameters, existing complications and treatment. RESULTS: According to this study, the yearly incidence of type 2 diabetes in Belgium is 231 per 100,000 inhabitants. Though the biochemical parameters (BMI, HbA1c, serum cholesterol and triglycerides) do not differ in the three regions of the country being investigated, there are considerable differences in the treatment prescribed for diabetes and its co morbidity. In Flanders, treatment is usually started with sulphonylurea, in the Walloon provinces with biguanides. Hypolipaemic treatment is also started more frequently in the latter region. In Flanders, hypertension is registered in 51.4% of the newly diagnosed patients with diabetes, which is higher than in the other regions. However, no difference is noted between the different regions in the number of patients with diabetes who are treated for hypertension. CONCLUSION: A network of sentinel physicians, taking part in voluntary registration, can be helpful in calculating the incidence of a health problem, in particular type 2 diabetes in general practice. The sentinel net can also serve as an instrument for describing patient characteristics and showing how physicians deal with health problems. The regional differences in medical approach that are described require further investigation, mainly with respect to their impact on the patients' outcome. PMID- 12086093 TI - Epidemiology of antibiotic resistance of human nontyphoidal salmonellae in Greece during an 8-year period (1990-1997). AB - The susceptibility patterns of 1027 nontyphoidal Salmonella strains of human origin, isolated in Greece between 1990 and 1997, were determined by broth microdilution. From 1990 to 1995, the overall incindence of resistance for both Salmonella enteritidis and S. typhimurium increased. From 1996 onwards, a decrease was observed, which was more evident for S. enteritidis. Regarding the other examined serotypes a substantial proportion of resistant isolates was found only for S. Virhow and S. PMID- 12086094 TI - Risk factor profile for atherosclerosis among young adults in Israel--results of a large-scale survey from the young adult periodic examinations in Israel (YAPEIS) database. AB - Assessing the prevalence of relevant risk factors among young adults is a critical step in the process of preventing atherosclerotic cardiovascular diseases (ASCVD) later in life. The Israel Defense Force Periodic Health Examination Center performs a routine check-up for subjects aged 25-45 years. Medical history, physical examination notes, laboratory results and ECG tracings are recorded, computerized and processed to form the Young Adults Periodic Examinations in Israel (YAPEIS) database. Data representing 31,640 subjects (27,769 males and 3871 females) examined between the years 1991-1999 were analyzed. The prevalence of documented risk factors for ASCVD were evaluated. The results of all parameters were graded categorically as low, moderate or high and the Framingham risk score was calculated. Fifty-one percent of the study participants were found to be overweight (body mass index > or = 25 kg/m2), 8.5% had high systolic blood pressure and 14.6% had high diastolic blood pressure. The prevalence of hypercholesterolemia and hyperglycemia was found to be 44.7 and 9.7%, respectively. Thirty-two percent of the subjects smoked cigarettes, and 76.7% reported not performing any routine physical activity. Furthermore, 31.8% had a Framingham score indicating a greater than 5% risk for developing a coronary event within the next 10 years. As expected, the prevalence of these risk factors increased with age and were found to be less frequent among females. Thus we conclude that many young Israeli adults hold significant risk factors for future ASCVD. Many of these risk factors are modifiable, and risk behavior is often amenable to alteration. Awareness to the high prevalence of risk factors among young adults should spark vigorous health-promotion programs as well as screening, education, and interventional measures aimed at altering the expected outcome of future ASCVD. PMID- 12086095 TI - Use of poisson regression and box-jenkins models to evaluate the short-term effects of environmental noise levels on daily emergency admissions in Madrid, Spain. AB - The relationship between environmental factors and hospital admissions has usually been analysed without taking into account the influence of a factor closely related to traffic in big cities, that is, environmental noise levels. We analysed the relationship between environmental noise and emergency admissions, for all causes and specific causes in Madrid (Spain), for the study period 1995 1997, using two statistical methods for the analysis of epidemiological time series data: Poisson autoregressive models and Box Jenkins (ARIMA) methodology. Both methods produce a clear association between emergency admissions for all and specific causes and environmental noise levels. We found very similar results from both methods for all and circulatory causes, but slightly different for respiratory causes. Around 5% of all emergency admissions can be attributed to high noise levels, with a lower figure for specific causes. Current levels of environmental noise have a considerable epidemiological impact on emergency admissions in Madrid. A reduction of environmental noise levels could be accompanied by a possible reduction in the number of emergency admissions. PMID- 12086096 TI - Work-related road accidents in France. AB - A descriptive analysis of all victims of road accidents recorded by the police in 1997 was carried out, distinguishing accidents occurring in the course of work, while commuting and for private reasons. The analysis concerned 105,816 drivers from 14 to 64 years of age; among these, 9.9% were injured during the course of work and 18.6% while commuting. The gender-ratio was maximal for accidents during the course of work (SR = 5.5) and minimal while commuting (2.1). Accidents in the course of work were generally less serious than the two other types aforementioned. Compared with the incidence rates for labourers, those of self employed trades and services (SETS) and those of employees and executives were higher for accidents incurred during the course of work and lower for those incurred during commuting. Professional drivers had the highest risk during the course of work. Accidents while working were most serious for SETS and professional drivers. Although it was impossible to take into account the mileage travelled, this study points out the real share of work-related road accidents and shows the need to develop and adapt prevention to the groups at risk that we have identified. PMID- 12086097 TI - Epidemiology of childhood bacterial meningitis in Poland. Incidence of bacterial meningitis with special reference to Haemophilus influenzae type b among children 0-59 months old in the former Kielce and Bydgoszcz districts in Poland in 1998 1999. AB - Population based surveillance was undertaken to assess the incidence of meningitis caused by Haemophilus influenzae type b in children 0-59 months old in Kielce and Bydgoszcz districts in Poland in 1998 and 1999. The cases were prospectively identified in pediatric and neuroinfection wards of local hospitals where all cases of children with suspected meningitis are referred in both districts. The mean annual incidence of meningitis caused by Haemophilus influenzae type b in children 0-59 months old in Kielce district during the study period was estimated at 3.1 per 100,000 per year (10.3% of cases of bacterial meningitis with confirmed etiology). In Bydgoszcz district, the annual incidence was 9.7 per 100,000 (50% of confirmed cases). These estimations are lower than reported in most Western European countries before the immunization against Hib was introduced. Small numbers of Hib vaccinations reported from both districts do not seem likely to have influenced the data significantly. PMID- 12086098 TI - Lack of association between intussusception and oral polio vaccine in Cuban children. AB - Two exploratory investigations found an increased risk of intussusception after oral polio vaccine (OPV). A large, national, population-based study was undertaken in Cuba to investigate a possible association. Three hundred and thirty-five cases of intussusception in children under 2 years of age occurring in 1995-2000 were identified and their OPV records retrieved. The relative incidence (RI) of intussusception in defined periods up to 42 days after OPV in children under 1 year was estimated using the self-controlled case series method, controlling for age and season. The RI was not significantly raised in any of the time intervals examined within the 0-42 day period after OPV. For the period 0-42 days as a whole the RI was 1.11, 95% CI 0.74-1.67. This study does not support the hypothesis that OPV causes intussusception. PMID- 12086099 TI - Apolipoprotein E genotype, lipid levels and coronary heart disease in a Polish population group. AB - Genotype of apolipoprotein E has been identified in a group of randomly selected Polish subjects participating in a cross-sectional study performed within the POL MONICA Program, the part of international study WHO-MONICA Project. The investigated group consisted of 170 persons, 92 males and 78 females aged 41-69 years (mean age 62.0+/-5.11). The observed frequency of apolipoprotein E alleles was: epsilon2 - 7.6%, epsilon3 - 81.8% and epsilon4 - 10.6%, which was similar to frequencies in the neighbouring European countries. Statistically significant lower means of total cholesterol (TC) and of low density lipoprotein cholesterol (LDL-C) levels in epsilon2 carriers and higher means of TC, of LDL-C and of triglycerides in epsilon4 carriers were observed as compared with noncarriers of respective alleles. Some nonlipid cardiovascular risk factors (hypertension (HT) and obesity) and coronary heart disease (CHD) showed a tendency to lower prevalence in the epsilon2 allele carriers as compared to noncarriers. In the epsilon4 allele carriers a tendency to higher prevalence of HT, but not of CHD was observed as compared to noncarriers of this allele. PMID- 12086100 TI - American cutaneous leishmaniasis as a rare imported disease in Europe: a case report favourably treated with antimonial derivatives. AB - A rare case report of American leishmaniasis imported to Italy after a prolonged journey in Brazil is described, and discussed according to the recent epidemiologic, diagnostic, and clinical literature evidence. Since a delayed diagnosis and treatment of American leishmaniasis outside endemic areas is common (due to a low clinical suspicion), careful and timely consideration of tropical infections is warranted in the differential diagnosis of cutaneous lesions found in a European traveller. PMID- 12086101 TI - Temperature resistant Nocardia biovarieties in Tucuman, Argentina? AB - Eighty-six regional strains of the pathogenic Nocardia species isolated from soil and human mycetoma were tested for their response to different incubation temperatures and for their tolerance to different temperatures. The aim was to assess whether growth temperature and tolerance to elevated temperatures are valuable criteria for the differentiation of pathogenic species of local strains based on the results obtained from a large number of strains. The results showed that 75.34% of all N. brasiliensis isolates from both sources grew at a temperature higher than 37 degrees C. 20% of the mycetoma strains and 11.32% of those from soil grew at 45 degrees C. 98.1% of N. brasiliensis from soil and 55.0% of the mycetoma strains tolerated 50 degrees C for 8 h and many isolates from both sources endured this temperature for an even longer time and tolerated yet higher temperatures. Both properties (growth temperature and temperature tolerance) are used to identify N. asteroides complex (N. farcinica) and N. otitidiscaviarum, and according to our results they are not suitable to differentiate regional strains of this species. The N. asteroides strains assayed showed an ability to grow at and tolerate elevated temperatures superior to those belonging to the other species. Although adaptation of local N. asteroides and N. otitidiscaviarum strains to temperature is important, it is more significant for N. brasiliensis, because this species is predominant in the Tucuman soil and responsible for the major number of diseases in the area. PMID- 12086102 TI - Yeasts isolated from blood and catheter in children from a public hospital of Sao Paulo, Brazil. AB - The great majority of nosocomial fungal infections, especially fungemias, are caused by yeasts, mostly of the genus Candida. In addition, such infections may be associated with intravascular catheters. In this study, 80 yeast strains were isolated from hospitalized children, being 59 from blood cultures and 21 from vascular catheter cultures. The prevalent species in both blood and catheter was C. parapsilosis (32.2% and 48.9%, respectively), followed by C. albicans (16.9% and 28.6%, respectively). Concerning enzyme production, 78.8% of the 80 isolates presented strong proteolytic activity but 78.8% showed no phospholipase activity. We also detected two prevalent "killer" biotypes: 511 and 888. Additionally, in five patients, it was possible to observe that the yeast species, "killer" biotype and proteolytic and phospholipase activity of blood and catheter were similar. In view of this, we suggest a transmission of nosocomial yeast infection from catheter to blood. PMID- 12086103 TI - The role of ascorbic acid role in the differentiation of sclerotia in Sclerotinia minor. AB - Sclerotinia minor in culture produces ascorbic acid in levels dependent on oxidative growth conditions and stage of development. During differentiation reduced/oxidized ascorbate ratio decreased by 12 and 6 fold at high and low oxidative stress, respectively. Exogenous ascorbate caused a concentration dependent decrease of oxidative stress (lipid peroxidation), inhibition of sclerotial differentiation (up to 100%) and delay of differentiatlon (up to 10 days). Ascorbic acid may be produced to help the fungus reduce oxidative stress during growth. The data of this study support our theory proposing that oxidative stress is the inducing factor of sclerotial differentiation in fungi. PMID- 12086104 TI - Effects of neem leaf extract on production of aflatoxins and activities of fatty acid synthetase, isocitrate dehydrogenase and glutathione S-transferase in Aspergillus parasiticus. AB - The relationship between the activities of 3 cytosolic enzymes with aflatoxin biosynthesis in Aspergillus parasiticus cultured under different conditions has been investigated in order to find out the role of each enzyme in aflatoxin biosynthesis. Basically the activity of isocitrate dehydrogenase (IDH) was higher in non-toxigenic strains as compared to its counterpart toxigenic fungi (p < 0.05). In contrast, the activities of fatty acid synthase (FAS) as well as glutathione S-transferase (GST) were higher (P < 0.05) in toxigenic strains than that of the non-toxigenic fungi. Aflatoxin production was inhibited in fungi grown in presence of various concentrations of neem leaf extract. Aflatoxin was at its lowest level (>90% inhibition) when the concentration of neem extract was adjusted to 50% (v/v). No significant changes in FAS and IDH activities were observed when aflatoxin synthesis was under restraints by neem (Azadirachta indica) leaf extract. During a certain period of time of culture growth, when aflatoxin production reached to its maximum level, the activity of FAS was slightly induced in the toxigenic strains fed with a low concentration (1.56% v/v) of the neem leaf extract. At the time (96 h) when aflatoxin concentration reached to its maximum levels, the activity of GST in the toxigenic fungi was significantly higher (i.e., 7-11 folds) than that of non-toxigenic strains. The difference was highest in mycelial samples collected after 120 h. However unlike FAS and IDH, GST was readily inhibited (approximately 67%) in mycelia fed with 1.56% v/v of the neem extract. The inhibition reached to maximum of 80% in samples exposed to 6.25-12.5% of the extract. These results further substantiate previous finding that there is a positive correlation between GST activity and aflatoxin production in fungi. PMID- 12086105 TI - Cloning and expression of pkaC and pkaR, the genes encoding the cAMP-dependent protein kinase of Aspergillus fumigatus. AB - This report describes the cloning and expression of both subunits of PKA in the opportunistic fungal pathogen Aspergillus fumigatus. The predicted translation product of the regulatory subunit, pkaR, is defined as a type II regulatory subunit. The gene encoding the A. fumigatus catalytic subunit, pkaC, contains the conserved kinase and activation domains that are characteristic of PkaC proteins. Both subunit mRNAs are expressed throughout the asexual life cycle of A. fumigatus. Message levels of pkaR and pkaC are higher during co-cultivation with alveolar epithelial cells than during culture alone. PMID- 12086106 TI - Dustborne fungi in large office buildings. AB - Fungi are ubiquitous in our daily environments. However, their effects on office workers' health are of great interest to many environmental health researchers. Dust has been considered an important reservoir of indoor fungi from which aerosolization and exposure could occur. We have examined the characteristics of dustborne fungal populations recovered from floors and chairs in office buildings. We investigated twenty-one offices in four office buildings in Boston, MA over a year beginning May 1997. We conducted intensive environmental sampling every six weeks to measure culturable dustborne fungi from floors and chairs, surface dust levels and water activity in carpeting. Carbon dioxide, temperature, and relative humidity were monitored continuously. Concentrations of total dustborne fungi recovered from floors were positively related to carbon dioxide (beta = 0.00064; p-value = 0.0002) and temperatures between 20 and 22.5 degrees C (p-value = 0.0026). Also, total fungal concentrations in floors gradually increased over the year (p-value = 0.0028). Total fungi recovered from chairs varied significantly by season (p-value < 0.0001), highest in September and lowest in March, and were positively correlated with dust loads in floors (beta = 0.25; p-value < 0.0001). We used principal component analysis (PCA) to reduce various observed fungal species to fewer factors. Six groups (PCA factors) were obtained for dustborne fungi recovered from both floors and chairs. The models of the first PCA factors for both floors and chairs were similar to those for total fungal concentrations. The results of this study provide essential information to further evaluate the effects of dustborne fungi on office workers' health. PMID- 12086107 TI - Neurophysiological and kinesiological aspects of spastic gait: the need for a functional approach. PMID- 12086108 TI - Correlation between tremor parameters. AB - The relationship between clinically scored tremor and electromyographic measurements of tremor amplitude is not clear and few reports have examined correlations between different tremor parameters. The aim of this study was to evaluate the correlations between different tremor parameters, clinical tremor scoring, age and disease duration. A total of 525 patients with different types of tremor were included in this study. Tremor in various limb positions was graded according to the Webster Tremor Scale. Surface electromyographic recordings of the limb found, in each position, to be most involved were performed in all the patients. The Spearman rank correlation coefficient was used to analyse the relationship between clinical and electromyographic parameters, while the Pearson correlation coefficient was used to analyse the relationship between the electromyographic parameters. Our results revealed moderate correlation between tremor amplitude as measured by electromyography and Webster tremor scores. The correlation between different tremor parameters revealed that low frequency tremors have higher amplitude, and longer burst duration, interval between bursts and missing interval. As the disease progresses, the tremor amplitude increases, while the frequency decreases. A strong positive correlation emerged between corresponding tremor parameters in different limb positions. This suggests that the tremor in all limb positions is generated by the same oscillator. In conclusion, a good correlation was found between clinically scored tremor and EMG measurements of tremor amplitude. The electromyographic examination is a useful tool for researching tremor mechanisms. PMID- 12086109 TI - Towards objective quantification of the Tinetti test. AB - The Tinetti test is a widespread test for assessing motor control in the elderly, which could also be usefully applied in neurology. At present it uses a qualitative measurement scale. As a first step towards its objective quantification, trunk inclination was measured during the test by two inclinometers and quantified by descriptive parameters. The 95th or 5th percentiles of parameter distributions in normal subjects (no.=150) were taken as limits of normality, and parameters computed on 130 institutionalised elderly people were compared to these limits, to test the parameters' discriminatory power. The distributions of many parameters were statistically different in normal subjects and patients. These results suggest that this approach is a promising tool for objective evaluation of the Tinetti test. PMID- 12086110 TI - Electromyographic mixed nerve and cutaneous silent period in evaluating the A delta fibres in a patient with hereditary sensory-autonomic neuropathy. AB - The aim of this study was to evaluate A-delta fibre function in a patient with hereditary sensory-autonomic neuropathy (HSAN). We used the mixed and cutaneous silent period techniques in addition to a conventional electromyographic investigation in a patient with type 2 HSAN, a rare disease characterised by wide spread sensory and variable autonomic dysfunction caused by incomplete development of sensory and autonomic neurons. Whereas the stimulation of one digital nerve did not show any evidence of silent period in either the left or the right hand, the simultaneous stimulation of two digital nerves, as well as the stimulation of a mixed nerve, revealed a measurable delayed and shortened silent period. These data suggest that a spatial summation mediated by A-delta fibres was required for generation of the silent period in this patient and that combining the CSP and MNSP may be of practical use in evaluating impairment of the small myelinated fibres. PMID- 12086112 TI - Antioxidant and cholinergic neuroprotective mechanisms in experimental parkinsonism. PMID- 12086111 TI - Cervicogenic headache: the importance of sticking to the criteria. AB - Cervicogenic headache (CEH) is a headache, but its origin is in the neck. Recently, two cases of intracranial tumour, which unfortunately were originally diagnosed as CEH, were published. The authors felt that this sequence of events demonstrates the insufficiency of the current CEH criteria. We--on the other hand -feel strongly that, on the contrary, this small challenge may have demonstrated the robustness of the CEH criteria. The criteria of CEH were actually not fulfilled at any time. We marshal our arguments in support of this view and concentrate on one of their cases (no. 1). PMID- 12086113 TI - Functional neurology at the dawn of the third millennium: the translation of neuroscience into neurology. PMID- 12086114 TI - VIII Antonio Arrigo award. Fatigue and sport. PMID- 12086115 TI - Response shift in the measurement of quality of life in hearing impaired adults after hearing aid fitting. AB - REASON FOR THE STUDY: Response shift is the change in the meaning of one's self evaluation of a target construct, like quality of life (QOL). The objective of this study was to investigate whether response shift in the measurement of generic and specific QOL occurred in persons with a relatively mild health condition. For this purpose hearing impairment was used as a research model. MAJOR FINDINGS: Response shift effects were observed in the scores on the dimensions of hearing related QOL. In the scores on overall hearing related QOL, and in the scores on the generic control items, no response shift occurred. CONCLUSIONS: This study showed that response shift effects can take place in a relatively mild condition as well. The occurrence of response shift in QOL ratings over time could have large implications for the estimation of the effectiveness of medical interventions and for the use of these estimations in cost-effectiveness analyses. After a successful treatment the conventional change could be an underestimation of the effectiveness of the treatment, although it has also been argued that psychological adaptation is a welcome capacity of human beings, and that then-test changes do no justice to this capacity. PMID- 12086116 TI - The impact of anxiety, depression and living alone in chronic obstructive pulmonary disease. AB - OBJECTIVE: This prospective longitudinal study examined whether quality of life as measured by the disease specific chronic respiratory questionnaire (CRQ) and living alone, at the time of prescription of long-term oxygen therapy (LTOT), impacted on the mortality of patients with severe chronic obstructive pulmonary disease (COPD). METHODS: One-hundred and fifty-seven patients, (male:female, 74:83) with severe COPD, aged under 80 years completed the CRQ at the time of prescription of LTOT. Demographic and physiological parameters were also collected. The patients were followed in outpatient clinics after commencing LTOT. RESULTS: Kaplan-Meier analysis of the data demonstrated a relationship between the emotional function score of the CRQ at the commencement of LTOT and survival on LTOT for female patients. Living with a partner was associated with an additional 12 months of life. When known physiological predictors of survival, the CRQ dimensions and living alone were included in a Cox's proportional hazard model, emotional function, body mass index and forced vital capacity were independent prognostic indicators for females, and arterial partial pressure of oxygen for males. CONCLUSION: Our results suggest that poor emotional functioning of female patients with severe COPD at the time of prescription of LTOT is associated with increased mortality. PMID- 12086117 TI - Quality of life of patients with Fabry disease. AB - This paper is the first of its kind to study the impact of Fabry disease (FD) in affected males, and shows that FD is associated with a significant decline in several domains. Using the medical outcomes study (MOS) SF-36 and a FD-specific questionnaire, we compared the observations found among these patients with that obtained for the general US population and other chronic disease states, including Gaucher disease (GD) (another lysosomal storage disorder), end-stage renal disease, stoke and AIDS. Patients with FD have a score profile most similar to patients with AIDS. In comparison with patients with GD, Fabry patients score substantially lower across all domains. Using simple linear regression, potential predictors of health-related quality of life (HRQOL) for Fabry patients were also determined. As in the general population, stroke, cardiac problems and renal disease lead to substantial decrement in HRQOL. In addition, two disease specific symptoms (acroparesthesia and anhidrosis) and pain are also predictors of decreased quality of life. Currently, no specific therapy for FD exists. As enzyme therapy for FD becomes increasingly available, it will be interesting to evaluate the therapy's impact on the quality of life of patients. PMID- 12086118 TI - Treatment and quality of life in patients with chronic obstructive pulmonary disease. AB - Treatments administered to patients with chronic obstructive pulmonary disease (COPD), especially when used in multiple combinations, are not free of interactions and side effects that can potentially impair health-related quality of life (HRQL). We studied HRQL and its relationship with treatment in a group of 441 patients with stage II or III COPD (age: 66.6 (SD: 8.3) years; FEV1: 32.4% (SD: 8.1%)) using the St George's Respiratory Questionnaire (SGRQ) and the 12 item short form (SF-12) Health Survey. The most prescribed drugs were ipratropium bromide (87.5%), inhaled corticosteroids (69.4%) and short-acting beta-2 agonists (64.9%). Patients with stage III of the disease were receiving more drugs, particularly short-acting beta-2 agonists (p = 0.002) and inhaled corticosteroids (p = 0.031). The use of theophyllines was associated with a worse total SGRQ score (beta = 4.49; p < 0.001), although this negative association decreased with advanced age. A trend towards worse SGRQ scores was observed with the use of high doses of long-acting beta-2 agonists (beta = 3.22; p = 0.072). Patients receiving three drugs or more presented worse total SGRQ scores than patients receiving fewer drugs (beta = 6.1, p < 0.001; and beta = 7.64, p < 0.001, respectively). These findings suggest that the use of multiple drugs in the treatment of patients with COPD is associated with worse total SGRQ scores. The effect of drugs, their dosages and associations with other drugs on HRQL merit further research. PMID- 12086120 TI - Discriminant properties of commonly used quality of life measures in heart failure. AB - Health-related quality of life (HRQL) instruments have been used to measure HRQL in heart failure patients, but how different instruments compare in the same groups of patients is not known. The purpose of this study was to compare the reliability and validity of three HRQL measures in 211 heart failure patients recruited from clinics affiliated with an urban hospital. Two disease-specific instruments, the chronic heart failure questionnaire (CHQ) and the living with heart failure questionnaire (LHFQ), and one generic instrument, the short-form 12 (SF-12), were administered. Patients reported moderate to low HRQL scores. Floor or ceiling effects were noted in the disease-specific instruments. Internal consistency reliabilities of the CHQ and LHFQ were satisfactory. Construct, convergent, and discriminant validity were supported for each instrument. Each scale and subscale, except for the SF-12 mental component scale, differentiated between patients with New York Heart Association (NYHA) class I, II, and III plus IV; the LHFQ physical subscale was the only measure to differentiate between patients with NYHA class III and IV. All three instruments were satisfactory for measuring HRQL, but the disease-specific instruments were preferable to the generic instrument. The decision of which instrument to use depends on the purpose of the study. PMID- 12086121 TI - Translation and validation of the Amsterdam preoperative anxiety and information scale (APAIS) for use in Japan. AB - Preoperative anxiety is a very common phenomenon that adversely affects a patient's physical and psychological outcome. To evaluate subjective complaints such as anxiety, use of a validated psychological instrument is essential. In this study, we translated, culturally adapted and validated the Amsterdam preoperative anxiety and information scale (APAIS) for use in Japan. One hundred and twenty-six preoperative patients participated in this cross-sectional study. The psychometric properties of the APAIS evaluated by factor analysis, correlation with the state-trait anxiety inventory (STAI) and Cronbach's alpha were very similar to those of the original Dutch and English versions. Female gender and a high information requirement were associated with high preoperative anxiety. In conclusion, the Japanese version of the APAIS is a valid and reliable instrument for screening Japanese patients for preoperative anxiety and information requirements. Female gender and high information requirement may be associated with high preoperative anxiety. PMID- 12086119 TI - The development of a new site-specific measure of quality of life for breast problems: the Cardiff breast scales. AB - The management of women presenting to primary care with symptoms of breast disease is of increasing interest given recent organisational changes aimed at improving accuracy and speed of referrals. As part of a randomised controlled trial, 1063 women were recruited following a primary care consultation for a variety of breast-related problems. In the absence of a suitable outcome measure for such women, a site-specific instrument was developed to complement a generic quality of life scale (SF-36). Items were generated using key informant interviews with health professionals. Draft scale items were piloted using a postal questionnaire and subsequent patient debrief interviews. A sample of respondents were also sent the same questionnaire I month later to assess test retest reliability. Across the whole sample (n = 848), three factors were identified: 'general well-being', 'concerns' and 'relationships'. These factors accounted for 60% of total variance. Evidence of scale validity, reliability and responsiveness are reported for this new outcome measure for use in women presenting with breast problems. PMID- 12086122 TI - Reliability and validity of the sexual life quality questionnaire (SLQQ). AB - The sexual life quality questionnaire (SLQQ) was developed to evaluate sexual quality of life (QOL) and satisfaction with treatments for erectile dysfunction among patients and their sexual partners. This paper describes the development of the instrument and reports its psychometric properties as observed in two studies involving patients under treatment for erectile dysfunction. The instrument consists of 16 items, 10 of which deal with dimensions of sexual QOL that can be summed to a sexual QOL scale measure. The remaining six items comprise a scale measuring satisfaction with treatment dimensions. The composite sexual QOL and treatment satisfaction scales met established psychometric goals overall and within select subgroups (length of time quartiles, patient/partner). A significant correlation between the treatment satisfaction scale score and the patients' responses to a question asking their likelihood of selecting the method for continued treatment (r: 0.89) showed the measure to be a good indicator of treatment preference. Finally, there were significant differences in sexual QOL scale scores between screening and first treatment, indicating the instrument was responsive and able to detect changes in sexual QOL. PMID- 12086123 TI - The impact of the Chi-Chi earthquake on quality of life among elderly survivors in Taiwan--a before and after study. AB - This paper examines the impact of the Chi-Chi earthquake, which hit central Taiwan on September 21, 1999, on the quality of life among the elderly survivors. The 28-item Taiwanese-adapted brief version of the World Health Organization's quality of life questionnaire (WHOQOL-BREF) was used to measure quality of life in four domains: physical capacity, psychological well-being, social relationships, and environment. These measures were coincidently collected in a separate study from 368 subjects aged 65 and older in the affected area shortly before the earthquake. Of these subjects, 268 were interviewed in a follow-up assessment 12 months after the earthquake. Linear mixed models were applied to investigate how quality of life in each of the four domains changed from the pre earthquake assessment to 12 months after the earthquake, and how these changes depended on the level of damage to residences. In conclusion, elderly survivors tended to report lower quality of life in physical capacity, psychological well being, and environment 12 months after the earthquake than at the assessment prior to the earthquake, regardless of the level of damage to their residences during the earthquake. However, those whose residences completely collapsed during the earthquake reported a higher quality of life in social relationships while others reported the opposite. PMID- 12086124 TI - Identification of quality of life concerns of patients with obstructive sleep apnoea at the time of initiation of continuous positive airway pressure: a discourse analysis. AB - BACKGROUND: Obstructive sleep apnoea syndrome (OSAS) is a common condition with multiple symptoms dominated by daytime somnolence. Thus many worries and concerns of patients remain hidden. Treatment by nasal continuous positive airway pressure (CPAP) can be imposing for the individual. An analysis of the freely expressed concerns of such patients is required. OBJECTIVE: To seek an in-depth analysis of how patients live with sleep apnoea by allowing them an open discourse and analysing the text of their statements. DESIGN: A trained psychologist conducted semi-directive interviews with patients attending a pulmonary rehabilitation and convalescent unit around the themes of sleep, health and treatment. An analysis of content and of discourse was carried out by textual analysis and by propositional analysis of discourse (PAD) with the aid of dedicated computer programs (Tropes, Sphinx Lexica). RESULTS: Thirty patients with severe sleep apnoea were interviewed of whom 15 were initiating treatment with CPAP. Patients spoke of abnormal fatigue (22 mentions) and somnolence (21 times). Many have problems with obesity (25 instances), snoring related problems (12). There were 30 mentions of depression with a relationship to alcohol and anti-depressives. Twenty six times the theme of nocturnal waking was raised. There were many instances of problems with CPAP (nasal mask and noise problems raised 21 times). Patients have problems with relationships and sex because of OSAS. Other concerns were loss of memory and fear of dying. CONCLUSION: In a non-directed conversation OSAS patients express concerns not revealed in the standard medical paradigm and such concerns should be addressed in assessing treatment or evaluating quality of life (QOL). PMID- 12086125 TI - Effect of acetaldehyde on acute tolerance and ethanol consumption in drinker and nondrinker rats. AB - OBJECTIVE: Acetaldehyde (AcH) has been shown to have aversive or reinforcing actions in relation to ethanol consumption. We have previously observed that a pharmacological dose of AcH (50-150 mg/kg) administered intraperitoneally (i.p.) produced a dose-dependent flavor aversion in low-ethanol drinker (UChA) rats, whereas high-ethanol drinker (UChB) rats appeared to be insensitive to AcH. Both strains of rats differ innately in their brain aldehyde dehydrogenase (ALDH) activity and in their capacity to develop acute tolerance to ethanol. The present study evaluates the effects of AcH, in UChA and UChB rats, on rats' behavior, their voluntary ethanol consumption and the development of acute functional tolerance to motor impairment induced by a dose of ethanol. METHOD: Subjects were rats selectively bred for high (UChB; n = 48) and low (UChA; n = 40) voluntary ethanol consumption. Rats were treated with either saline or doses of AcH (50 or 100 mg/kg, i.p.), and examined for effects on motor activity, on voluntary alcohol consumption with free access to a 10% alcohol solution and on acute tolerance to motor impairment induced by an ethanol dose (2.3 g/kg, i.p.), using the tilting plane test. RESULTS: AcH, 50 or 100 mg/kg, caused a dose-dependent loss of the righting reflex in UChA rats, whereas in UChB rats with same dose, a slight excitement was observed. There was significant increase of voluntary ethanol consumption in UChB rats (p < .001) 17 hours after the AcH injection, compared with saline-treated and control rats; in UChA rats, no change in voluntary ethanol consumption was produced. AcH produced a faster acute tolerance to-ethanol development in UChB rats (p < .001) as compared with saline-treated and control rats, and no change in acute tolerance in UChA rats. Acetaldehyde injection did not change total mitochondrial ALDH2 activity. CONCLUSIONS: These results show that, 17 hours later, treatment of rats with pharmacological doses of acetaldehyde alters the voluntary ethanol consumption and acute tolerance development in UChB but not UChA rats. One of the factors involved may be a different sensitivity to AcH in these rat strains. PMID- 12086126 TI - Alcohol abuse and dependence among U.S. college students. AB - OBJECTIVE: To estimate the prevalence of alcohol abuse and dependence among U.S. college students, and to identify characteristics associated with these diagnoses. METHOD: More than 14,000 students at 119 4-year U.S. colleges completed a questionnaire that included items corresponding to DSM-IV diagnostic criteria for alcohol abuse and dependence. Frequencies were computed, and correlations used to identify demographic, drinking and other variables associated with these diagnoses. RESULTS: 31% percent of students endorsed criteria for an alcohol abuse diagnosis and 6% for a dependence diagnosis in the past 12 months. More than two of every five students reported at least one symptom of abuse or dependence. Students who were heavy episodic drinkers were more likely than those who were not to have an alcohol disorder. Students who were frequent heavy episodic drinkers had 13 times greater odds for abuse and 19 times greater odds for dependence. One of every five heavy episodic drinkers was classified with dependence. Few reported seeking treatment since coming to college. Students from heavy drinking college environments were more likely to have abuse and dependence diagnoses. CONCLUSIONS: Many college students report behaviors and symptoms that meet the diagnostic standard for alcohol abuse or dependence. In addition to strengthening prevention programs, colleges should implement new strategies for screening and early identification of high risk student drinkers and ensure that treatment is readily available for those with alcohol disorders. PMID- 12086127 TI - The impact of current residence and high school drinking on alcohol problems among college students. AB - OBJECTIVE: This study examines relationships between type of (current) residence, heavy episodic drinking in high school and alcohol-related problems among college students. METHOD: The study participants were respondents in the 1993, 1997 and 1999 Harvard School of Public Health College Alcohol Study (CAS) surveys of students attending 119 4-year U.S. colleges. Based on responses from 6,525 (55.6% female) students in the 1993 CAS, an exploratory factor analysis of the alcohol problem items was specified in a confirmatory factor analysis framework based on a four-factor solution, and related to study variables. The 1993 data were cross validated with the 1997 and 1999 surveys. RESULTS: When compared with students living in single-gender dormitories, students living off campus with parents reported lower alcohol-related problem consequences and a higher probability of drinking/driving. Students residing off campus without parents, compared with students in single-gender dorms, reported a higher probability of drinking/driving. Associations between off-campus residence and probabilities for drinking/driving were mediated by frequency of driving. Students living in coed dormitories, when compared with students in single-gender dorms, incurred more problem consequences related to drinking but reported significantly lower probabilities associated with designated driving and drinking/driving. Heavy episodic drinking in high school was related to higher probabilities of problems on all outcome measures. CONCLUSIONS: The presence of direct and independent effects for both heavy drinking prior to college and high-risk environmental factors in collegiate drinking practices support targeted and diverse strategies for prevention activities. PMID- 12086129 TI - Reducing high-risk drinking by young Americans south of the border: the impact of a partial ban on sales of alcohol. AB - OBJECTIVE: Determine the effect of a partial sales ban on cross border drinking in Mexico. METHOD: On weekend evenings, thousands of youths (younger than 21 years) and young adults (21 to 25 years) residing in communities along the U.S. border cross over into Mexico to patronize all-night bars where the drinking age is 18 rather than 21 years and where the price of alcohol is considerably less than in the United States. On January 1, 1999, Juarez, Mexico, across the border from El Paso, Texas, implemented a 2 AM bar closing policy replacing the previous 5 AM closing time. Breath alcohol tests of pedestrians at the Juarez/El Paso border before and after the policy change were compared with a similar sample of pedestrians at the Tijuana, Mexico/San Diego, California border. RESULTS: At the Juarez/El Paso border, the total number of youths with positive BACs returning from Juarez after 3 AM when the bars were closed was reduced 89%, whereas the number returning between midnight and 3 AM remained unchanged. There was no change in either period at the Tijuana/San Diego comparison site. CONCLUSIONS: Early closing of the bars in Juarez reduced the number of youths returning after 3 AM to the United States with positive BACs. PMID- 12086128 TI - Motivational structure and alcohol use of university students across four nations. AB - OBJECTIVE: The ability of motivational structure and other variables to predict alcohol consumption was assessed in university students (N = 370; 244 women) in the Czech Republic, The Netherlands, Norway and the United States. METHOD: Motivational structure was assessed with the Motivational Structure Questionnaire (MSQ), which inquires about respondents' individual personal concerns in various areas of life and dimensions of their goal strivings related to the resolution of concerns. Alcohol-related problems were measured with the Short Michigan Alcoholism Screening Test. RESULTS: Factor analysis of MSQ indices yielded a two factor solution, one factor of which reflects an adaptive motivational structure marked by elements necessary for attaining psychologically satisfying resolutions of personal concerns. The other factor reflects a more maladaptive motivational structure marked by feelings of indifference about the achievement of personal goals. As hypothesized, MSQ Factor I was a significant inverse predictor of the amount of alcohol that students drank, but only among those who experienced alcohol-related problems. CONCLUSIONS: The similarity in results across culturally varied samples suggests the robustness of the relationships for understanding the motivational bases for alcohol use. PMID- 12086131 TI - The importance of a positive family history of alcoholism, parental rejection and emotional warmth, behavioral problems and peer substance use for alcohol problems in teenagers: a path analysis. AB - OBJECTIVE: The purpose of this study was to test a hypothetical model of alcohol problems in German adolescents. Among 180 offspring, family history of alcoholism, parenting styles, behavioral and emotional problems, peer-group characteristics, feelings of self-esteem, behavioral problems and psychiatric comorbidity of the parents were examined. METHOD: Data were generated from the Study of Health in Pomerania (SHIP), in which families were randomly selected if 12-18 year old biological offspring were members of the household; a smaller group of subjects was selected from local outpatient treatment centers. Members of 133 families, including 180 (50.6% male) offspring who were appropriate for the current analyses, received personal semistructured diagnostic interviews and several self-rating questionnaires. Analyses compared offspring with alcohol problems (AP; n = 40) and with no alcohol problems (NAP; n = 140), and used structural equation modeling to test a hypothetical model. RESULTS: The comparisons revealed that the AP group had significantly more behavioral problems (e.g., aggression/delinquency), more perceived parental rejection and less emotional warmth, a higher amount of alcohol consumption, were more likely to associate with substance-using peers and more often received a diagnosis of conduct disorder or antisocial personality disorder. Whereas the family history of alcoholism did not differ significantly between groups, parents of offspring with an alcohol use disorder had significantly more additional diagnoses on DSM IV Axis I. The evaluation of the model supported the importance of aggression/delinquency and association with substance-using peers for alcohol problems in people. An additional diagnosis in the parents was directly and indirectly (through aggression/delinquency) related to alcohol problems of the adolescents. CONCLUSIONS: The data indicate that alcohol problems in the offspring are associated with several domains of influence in their environment. Prospective studies measuring both biological and environmental factors using sufficient sample sizes will be needed for optimal understanding of the development of alcohol problems in youth. PMID- 12086132 TI - Beyond alcoholism: identifying older, at-risk drinkers in primary care. AB - OBJECTIVE: To evaluate the validity and reliability of two self-report instruments: the Alcohol-Related Problems Survey (ARPS) and its shorter version the Short ARPS (shARPS) that identify older persons whose use of alcohol alone or with their comorbidities may be placing them at risk for or causing them harm. METHOD: We compared the two measures against a "LEAD" (longitudinal evaluation done by experts employing all available data) standard among a sample of 166 drinkers aged 60 years and older in 10 internal medicine clinics. The LEAD standard included a medical record review, a clinical interview and a telephone interview with a collateral informant. We tabulated reasons the LEAD identified subjects as harmful or hazardous drinkers. We also compared the Alcohol Use Disorders Identification Test (AUDIT) and the Short Michigan Alcoholism Screening Test-Geriatric Version (SMAST-G) to the LEAD. RESULTS: Sensitivity and specificity of the ARPS and the shARPS as compared to the LEAD were 93% and 63%, and 92% and 51%, respectively. After minor changes were made in the scoring rules, specificity increased to 66% for both the ARPS and shARPS while sensitivity remained stable, 93% and 91%. Harmful and hazardous drinkers were most often identified because of alcohol use with comorbidities, symptoms, and medication use. Sensitivity and specificity of the AUDIT and the SMAST-G as compared to the LEAD were 28% and 100%, and 52% and 96%, respectively. CONCLUSIONS: The ARPS and shARPS are quite sensitive in identifying older drinkers with a spectrum of alcohol use disorders. They are more sensitive than the AUDIT and the SMAST-G in identifying older persons who may be at risk or experiencing harm as a result of their alcohol use and comorbidities. They also provide information on specific risks associated with alcohol use not obtained by other screening measures and may therefore facilitate interventions by busy clinicians to reduce such risks. PMID- 12086133 TI - Stability of alcohol consumption over time: evidence from three longitudinal surveys from the United States. AB - OBJECTIVE: To assess the stability of individual monthly alcohol consumption volume in three studies from the United States: The first National Health and Nutrition Examination Survey with three measurements, The Framingham Offspring and Spouse Cohort with four measurements, and the National Longitudinal Survey of Youth with six measurements. The results will aid in the interpretation of studies that assess consumption at one point only. METHOD: Five analyses measure (1) the amount of change between measurements, (2) the correlation between reported consumption, (3) patterns of later drinking by the baseline group, (4) mean consumption of abstainers when they drink and heavy drinkers when not drinking heavily and (5) the strength of past consumption in predicting present consumption. RESULTS: Correlations between measurements are high for adult samples 5 years apart or less but low for longer follow-ups and younger samples. Heavy drinkers are much less stable than abstainers or moderate drinkers. Abstainers at one time who drink at another drink much less than the mean. One time heavy drinkers are close to the consumption mean of the entire sample when not heavy but two or more time heavy drinkers drink more than the mean when not heavy. An autoregressive, rather than Markov, model of consumption change is supported. CONCLUSIONS: Aspects of both stability and change are found. Baseline measures of drinking groups are especially unreliable for younger samples, longer follow-ups and heavier drinkers. There appear to be important subgroups moving between abstention and light drinking and moderate and heavy drinking that can be identified only by multiple measurements or retrospective measures. PMID- 12086130 TI - Do adolescents affiliate with 12-step groups? A multivariate process model of effects. AB - OBJECTIVE: Research with adolescents has revealed salutary effects for 12-step attendance on substance use outcomes, but no studies have examined the effects of 12-step affiliation, or active involvement, beyond simple measures of attendance. Prior research with adults has shown that measures of affiliation are more predictive than measures of attendance. This study (1) assessed attributes that may influence 12-step attendance and affiliation; (2) tested whether 12-step affiliation in the first 3 months posttreatment possessed unique predictive power above that attributable to attendance alone; and (3) examined the extent to which motivation, coping and self-efficacy measured at 3 months mediated the relation between 12-step affiliation and substance use outcome in the ensuing 3 months. METHOD: Adolescent inpatients (N = 74, 62% female), who were aged 14-18 years (mean [SD] = 15.9 [1.19] years), were interviewed during treatment and at 3 and 6 months postdischarge. RESULTS: More severely substance-involved youth were more motivated for abstinence and more likely to attend and affiliate with 12-step groups. A high degree of collinearity between 12-step attendance and affiliation suggested that those attending were also likely to be those actively involved. As a consequence, affiliation did not predict outcome over and above frequency of attendance. Motivation was found to influence the relationship between 12-step affiliation and future substance use outcome. CONCLUSIONS: Given the widespread treatment recommendations for adolescent 12-step involvement, more study is needed to determine what kinds and what aspects of 12-step groups and fellowships are helpful to adolescent change efforts and what alternatives should be developed. PMID- 12086134 TI - Environmental contexts surrounding resolution of drinking problems among problem drinkers with different help-seeking experiences. AB - OBJECTIVE: This study investigated whether similar environmental contexts surround abstinent resolutions associated with different intervention experiences, including no assistance. METHOD: Participants were selected in a 3 x 2 design (25-30 per group, N = 167) according to their help-seeking experiences (no assistance, Alcoholics Anonymous [AA] only, treatment plus AA) and current drinking status (resolved abstinent [RA] for >2 years or nonresolved [NR] controls). Life events were assessed retrospectively over a 4-year period that spanned the 2 years before and 2 years after the initiation of stable abstinence by RA participants or over a matched interval for NR participants. Collateral or reliability interviews were conducted for 84% of the sample as checks on participant reports. RESULTS: Across help-seeking groups, RA participants reported decreased negative events and increased positive events from the pre- through the post-resolution period. Interventions, especially treatment, enhanced the postresolution improvements in RA participants' life circumstances. This pattern was absent among NR participants. CONCLUSIONS: The findings suggest that a similar molar environmental context surrounds resolutions achieved with and without interventions and that interventions are associated with enhanced improvements during maintenance. PMID- 12086135 TI - Effect of news coverage on the prevalence of drunk-driving behavior: evidence from a longitudinal study. AB - OBJECTIVE: To examine the proposition that antidrunk driving messages in the news media contributed indirectly to the decline in drunk driving over the past two decades through their impact on related policy making processes. METHOD: Time series regression techniques are applied to longitudinal data to examine the causal association between drivers' involvement in drunk-driving behavior, the volume of news coverage devoted to the drunk driving issue, and related policy making. RESULTS: Results show a significant contribution of news coverage to drunk-driving-related policy actions, which in turn are associated with a reduction in drunk driving among young and high-risk drivers. There was no evidence of a direct causal association between news coverage and change in drunk driving behavior. CONCLUSIONS: News coverage of alcohol-related risky behaviors seems to provide a cost-effective way of reducing the prevalence of these practices by attracting institutional attention and prompting related environmental changes. Future interventions may benefit from actively seeking to influence news coverage of risky behaviors. PMID- 12086136 TI - Concurrent and retrospective reports of alcohol consumption across 30, 90 and 366 days: interactive voice response compared with the timeline follow back. AB - OBJECTIVE: To compare near-concurrent and retrospective reports of alcohol consumption and to determine whether a simple adjustment function could be applied to the retrospective reports to correct any distortions in reporting. METHOD: Reports of daily alcohol consumption collected via an Interactive Voice Response (IVR) system were aggregated over 30, 90 and 366 days and compared with the corresponding retrospective reports gathered with the Timeline Follow Back (TLFB). Comparisons of the IVR and TLFB reports for five consumption variables were made for each time period for 33 men. RESULTS: Significant differences between reporting methods were found for four of the five variables tested. On average, the TLFB yielded an underestimate of alcohol consumption compared to the IVR. This underestimate associated with TLFB reports was not dependent on the length of the recall period. Regression analyses indicated that the relationship between the two modalities of reporting was dependent on the quantity of drinks reported, thus precluding the application of a simple, linear metric to the TLFB data to model near real-time daily data obtained through the IVR more accurately. CONCLUSIONS: The retrospective TLFB consistently and significantly underestimated alcohol consumption compared to the aggregated daily reports. This underestimate, however, was stable across the three reporting periods. With one exception, no simple linear function could be applied to the retrospective TLFB consumption reports to more accurately reflect the IVR data gathered daily. Evidence presented here also suggests that underreporting on the TLFB is not simply a matter of consciously attempting to conceal consumption. PMID- 12086137 TI - Substance-induced and independent major depressive disorder in treatment-seeking alcoholics: associations with dysfunctional attitudes and coping. AB - OBJECTIVE: This study examines the validity of the distinction between substance induced and independent major depressive disorder (MDD) among alcoholics utilizing variables theoretically linked to the etiology and course of depression: specifically dimensions of coping and dysfunctional attitudes about the self that can lead to extreme negative interpretations of life events. METHOD: Participants were 166 alcohol-dependent patients with elevated depressive symptoms. Comparisons were made between participants who met criteria for a current substance-induced major depressive episode and did not have a history of independent MDD (SIDD) and those who met criteria for a current major depressive episode and had either a past or current episode of independent MDD (IMDD). We also compared both of these groups to participants who have never met criteria for a depressive episode (NMDE). RESULTS: Depressive symptoms were correlated negatively with dimensions of adaptive coping and positively with maladaptive coping and dysfunctional attitudes. Women were overrepresented in the IMDD group, but the diagnostic groups did not differ significantly in other demographic characteristics or in measures of substance use. Participants with IMDD reported greater dysfunctional attitudes than those with NMDE and those with SIDD, who did not differ from each other. Frequency of antidepressive coping behaviors was lowest among those with IMDD, was significantly higher in those with SIDD and was highest among those with NMDE. CONCLUSIONS: Results suggest depressed alcoholics who have experienced episodes of depression independent of excessive substance use possess cognitive vulnerabilities and coping deficits that may render them prone to depression. PMID- 12086138 TI - Alcohol consumption, setting, gender and activity as predictors of injury: a population-based case-control study. AB - OBJECTIVE: A population-based case-control design was employed to quantify the risk of injury after consumption of alcohol as a function of setting, concurrent activity and usual drinking habits. METHOD: A total of 797 cases (66.6% men) and 797 controls (57.7% women) were interviewed. The response rate was 83% for eligible cases approached for an interview. Cases were injured patients from a hospital emergency department. Community controls used were each paired with a case on suburb of residence, and interviewed regarding their activities in the 6 hour period preceding their paired case's injury. RESULTS: Self-reported alcohol consumption was consistent with both medical records and breath-analyzer tests. Drinking any alcohol and using prescribed medication in the prior 6 hours were both associated with significantly increased risk of injury when controlling for demographic and setting variables. Use of illicit drugs (mainly cannabis) was associated with reduced risk of injury. Setting (e.g., recreational, work) and activity (e.g., sport, travel, work) variables were also independently associated with risk of injury. The risk of injury for women was significantly elevated for any consumption of alcohol; for men it was elevated only when consumption exceeded 90 grams. CONCLUSIONS: These data confirm earlier findings that risk of injury for women for a given level of consumption is greater than for men. They extend earlier findings by identifying significant setting, activity and drug use variables predictive of injury. In addition, when these latter variables are controlled, it is found that for women, but not for men, the risk of injury is significantly elevated even at low levels of alcohol intake. PMID- 12086139 TI - A comparison of alcohol-positive and alcohol-negative trauma patients. AB - OBJECTIVE: Hospital admission for an alcohol-related traumatic injury may offer a "teachable moment" to address a patient's alcohol problem. Although trauma teams provide a number of other health-related services, there may be characteristics of alcohol-positive victims that act as barriers toward providing alcohol counseling. The purpose of this study was to compare the characteristics and hospital outcomes of trauma patients who tested positive for alcohol at the time of hospital admission with those who did not. This information is useful for planning interventions and referrals for treatment. METHOD: The study was a retrospective comparison of alcohol-positive and alcohol-negative patients who were admitted for at least 48 hours to a Level-I trauma center. Data from 1,049 trauma victims (736 male, 742 alcohol-negative) were abstracted from clinical records. RESULTS: Several characteristics were found to be associated with alcohol-related injuries: being male, aged 40 years or less, having a toxicology screen positive for illicit drugs, lacking health insurance, being indigent and sustaining an injury related to violence. Alcohol-positive patients were also found to spend fewer days in a critical care unit, to be less likely to die and to be less likely to be transferred to another hospital than alcohol-negative patients, despite having similar injury severity. CONCLUSIONS: Patient characteristics suggest that there are obstacles to providing interventions and referrals by healthcare professionals for victims of alcohol-related injuries. Less expensive options that consider the demographic features of this patient population need to be developed as an alternative to expensive, professional interventions. PMID- 12086140 TI - Macropodid herpesvirus 1 encodes genes for both thymidylate synthase and ICP34.5. AB - Macropodid herpesvirus 1 (MaHV-1) is an unclassified alphaherpesvirus linked with the fatal infections of kangaroos and other marsupials. During the characterisation of the internal repeat region of MaHV-1, an open reading frame (ORF) encoding for thymidylate synthase (TS) gene was identified and completely sequenced. Southern blot analysis confirmed the presence of two copies of the TS gene in the MaHV-1 genome as expected. Computer analysis of the TS ORF showed it was 948 nucleotides in length. A putative polyadenylation signal was identified 17-22 bp inside the ORF implying a minimal or absent 3' untranslated region. The predicted polypeptide was 316 amino acid residues in length and contained the highly conserved motifs for folate binding and F-dUMP binding, typical of all TS enzymes. Interestingly, MaHV-1 TS polypeptide had highest similarity to the human TS polypeptide (81%) compared to the TS polypeptides of other herpesviruses (72 75%). Immediately upstream of the TS gene, a second ORF of 510 bp, encoding a polypeptide with 170 amino acid residues, was identified. The carboxyl domain of this MaHV-1 polypeptide shared 68% similarity to a 59 amino acid motif of human herpesvirus 1 ICP34.5, identifying it as the MaHV-1 ICP34.5 homologue. This is the first report of a herpesvirus that encodes for both TS and ICP34.5. PMID- 12086141 TI - Hepatitis B virus vector carries a foreign gene into liver cells in vitro. AB - Hepatitis B viruses (HBV) specifically target the liver, where they efficiently infect quiescent hepatocytes. Thus, HBV virus has potential to be used as vectors for liver-directed gene transfer. We constructed a new HBV-based vector system. It is composed of transfer vector for transferring a foreign gene, green fluorescence protein (GFP) gene, and a helper vector. When the transfer vector and the helper vector were cotransfected into HepG2 cells, the recombinant HBV (rHBV) particles were generated by trans-complementation between two vectors. The rHBV particles carrying the foreign gene were identified by the Southern blot assay. To test gene delivery and the transduction of the rHBV, we infected primary human hepatocytes and immortalized, HepG2 cells with rHBV in vitro. The results using fluorescence microscopy confirmed that the inserted GFP gene was successfully transferred and expressed both in primary human hepatocytes and HepG2 cells. PMID- 12086142 TI - Nucleotide sequence and phylogenetic analysis of Cucurbit yellow stunting disorder virus RNA 2. AB - The complete nucleotide sequence of Cucurbit yellow stunting disorder virus (CYSDV) RNA 2, a whitefly (Bemisia tabaci)-transmitted closterovirus with a bi partite genome, is reported. CYSDV RNA 2 is 7,281 nucleotides long and contains the closterovirus hallmark gene array with a similar arrangement to the prototype member of the genus Crinivirus, Lettuce infectious yellows virus (LIYV). CYSDV RNA 2 contains open reading frames (ORFs) potentially encoding in a 5' to 3' direction for proteins of 5 kDa (ORF 1; hydrophobic protein), 62 kDa (ORF 2; heat shock protein 70 homolog, HSP70h), 59 kDa (ORF 3; protein of unknown function), 9 kDa (ORF 4; protein of unknown function), 28.5 kDa (ORF 5; coat protein, CP), 53 kDa (ORF 6; coat protein minor, CPm), and 26.5 kDa (ORF 7; protein of unknown function). Pairwise comparisons of CYSDV RNA 2-encoded proteins (HSP70h, p59 and CPm) among the closteroviruses showed that CYSDV is closely related to LIYV. Phylogenetic analysis based on the amino acid sequence of the HSP70h, indicated that CYSDV clusters with other members of the genus Crinivirus, and it is related to Little cherry virus-1 (LChV-1), but is distinct from the aphid- or mealybug transmitted closteroviruses. PMID- 12086143 TI - Phosphorylation of the movement protein of cucumber mosaic virus in transgenic tobacco plants. AB - The 3a protein of Cucumber mosaic virus is essential for the cell-to-cell movement of the viral RNA through plasmodesmata. We have introduced an epitope peptide before the stop codon of the 3a protein and cloned the tagged ORF into a binary vector for Agrobacterium-mediated transformation. The established transgenic tobacco lines produced the 3a protein, which was specifically detected with anti-3a and anti-epitope antisera. Metabolic labeling and subsequent immunoprecipitation revealed that [32P]-orthophosphate was incorporated into the 3a protein. The phosphoamino acid analysis indicated that the 3a protein contained phosphoserine but not phosphothreonine or phosphotyrosine. This is the first demonstration of the 3a protein phosphorylation in planta. PMID- 12086144 TI - Cloning and regulation of the promoter of pseudorabies virus (TNL strain) glycoprotein E gene. AB - The nucleotide sequence upstream to the glycoprotein E (gE) gene of pseudorabies virus (PrV, TNL strain) was cloned from the genomic virus DNA by polymerase chain reaction (PCR) and its DNA sequences were determined. The DNA segment, which was supposed to contain the gE promoter, was subcloned into a chloramphenicol acetyltransferase (CAT) reporter gene and the resulting plasmid was named pgEp-B CAT. To examine the promoter function of this upstream sequence of gE gene, we transfected pgEp-B-CAT DNA into L-M cells and the promoter activity was analyzed by CAT assay. Results showed that our DNA fragment could exhibit promoter activity. Furthermore, we transfected L-M cells with pgEp-B-CAT for 48 h, then superinfected cells with pseudorabies virus, and performed CAT assay. It was found that PrV superinfection could slightly enhance the activity of gE promoter, suggesting that factors produced during viral infection could stimulate the promoter. To explore the possible mechanism of regulation at transcriptional level, the pgEp-B-CAT plasmid were cotransfected with eukaryotic vectors expressing viral regulatory proteins IE or EP0, and results indicated that the gE promoter was activated by IE protein whereas it was inhibited by EP0 protein. Moreover, the effect of exogenous IE or EP0 on the protein level of gE in PrV infected cells was examined; conclusion similar to that of CAT assay were obtained. PMID- 12086145 TI - Characterization of the p10 gene region of Anticarsia gemmatalis nucleopolyhedrovirus. AB - The Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) p10 gene region was cloned, sequenced and the putative p10 gene expression characterized by Northern blot analysis. Sequence analysis of the p10 gene region indicated the presence of two complete open reading frames (ORFs) of 713 and 281 nucleotides, which codes for polypeptides of 273 and 93 amino acids, with homology to the P26 and P10 proteins of baculoviruses, respectively. Two additional partial ORFs, coding for partial polypeptides of 110 and 146 amino acids, showed homology to the p22.2 gene of Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) and p74 genes of different baculoviruses, respectively. A small ORF of 224 nucleotides coding for a protein of 74 amino acids showed homology to the 3'-end of the early p94 gene of AcMNPV. A putative baculovirus very late promoter motif TAAG was identified in the 5'-non-translated region (5'-UTR) at position-54 upstream of the start codon. The consensus polyadenylation sequence AATAAA is present 146nt downstream of the termination codon and the p10 ORF is flanked by the p26 and p74 ORFs. Homology comparisons showed that the P10 protein of AgMNPV is most closely related (82% amino acid sequence identity) to the P10 from the Orgyia pseudotsugata nucleopolyhedrovirus (OpMNPV). Transcriptional analysis of the AgMNPV p10 gene showed that p10-specific transcripts could be detected late in infection. PMID- 12086146 TI - Molecular analysis of Cotton leaf curl virus-Sudan reveals an evolutionary history of recombination. AB - Monopartite begomoviral DNAs (2761 bp) were cloned and sequenced from field cotton, okra, and Sida alba, from Gezira, and field okra from Shambat. Comparison of the four apparent full-length begomoviral DNAs revealed 99.3-99.5% shared nucleotide (nt) identity, indicating that they are the same viral species, hereafter, referred to as Cotton leaf curl virus-Sudan (CLCuV-SD). Host range studies revealed that the field okra isolate of CLCuV-SD was whitefly transmissible from okra to okra, M. parviflora, and hollyhock, but not to cotton. In contrast, the cotton isolate of CLCuV-SD infected cotton and hollyhock, but not okra. The genome of CLCuV-SD encodes six open reading frames (ORFs), and was most closely related to other monopartite begomoviruses of the Eastern Hemisphere. CLCuV-SD shared highest nucleotide sequence identity (95.5%) with Okra enation virus (OkEV), but was distantly related (approximately 74% nt sequence identity) to begomoviruses isolated from cotton in Pakistan. While extensive genomic regions of CLCuV-SD and OkEV are highly conserved (approximately 99% nt identity), nt sequence identity of the V1 ORF encoding the coat protein was uncharacteristically low (87.9%), suggesting a history of recombination. An analysis conducted with Sawyer's GENECONV program support the recombination hypothesis, indicating that the V1 ORF and a small segment of the intergenic region of CLCuV-SD and OkEV were derived from other begomoviruses. As a BLAST analysis failed to identify a prospective extant source of either V1 ORF, the parental viruses serving as CP donors remain undiscovered or are extinct. PMID- 12086147 TI - Identification and characterization of the UL7 gene product of herpes simplex virus type 2. AB - We have raised a rabbit polyclonal antiserum against a recombinant 6x His-tagged herpes simplex virus type 2 (HSV-2) UL7 fusion protein expressed in Escherichia coli. The antiserum specifically reacted with a 33 kDa protein in HSV-1 and HSV-2 infected cell lysates, and was used to characterize the UL7 gene product of HSV 2. The UL7 protein was produced in the late phase of infection, and its synthesis was highly inhibited, but not abolished by the addition of acyclovir (ACV). The UL7 protein associated with extracellular virions and also with all types of capsids, including A, B, and C capsids, though the association seemed to be weak. Indirect immunofluorescence studies revealed that at 9 h postinfection, UL7 specific fluorescence was detected in part or all of the nucleus, and the specific fluorescence colocalized with the scaffold protein ICP35. However, at later times postinfection, the UL7 protein was mainly detected as a mass in a juxtanuclear cytoplasmic region. In addition, transmission immunoelectron microscopy (TIEM) confirmed the association of the UL7 protein with intracellular capsids and virions in HSV-2-infected cells. The HSV-2 UL7 protein contained a domain highly conserved in all herpesviruses, part of which exhibited a homology with domains in the fission yeast Schizosaccharomyces pombe DNA topoisomerase III. We discuss the possibility that the UL7 protein may play a supplementary role in the viral DNA cleavage/packaging process. PMID- 12086148 TI - Viral evolution and emerging viral infections: what future for the viruses? A theoretical evaluation based on informational spaces and quasispecies. AB - Emergence of new viruses is dependent on the intrinsic and extrinsic constraints exerting on viral evolution. Intrinsic constraints are semantic and grammatical in nature. They are analysed here in reference to Hamming's spaces, driving to a new interpretation of the evolution of the quasispecies of Manfred Eigen. Extrinsic constraints are relevant to the fact that viral evolution is always a co-evolution story, with two or three partners implicated (the viruses, their hosts and eventually their vectors). They imply that viral phylogenies are disconnected, and viruses constitute a polyphyletic system. A possible consequence is that potential viral families are already known, or at least are present in nature, in such a manner that the frames for future viral evolution are already determined and that the probability for the emergence of a new frame is nil. Nevertheless, the emergence of new pathogens in the existing frames remain possible. PMID- 12086150 TI - The effect of intracerebroventricular dopamine administration on the respiratory response to hypoxia. AB - Acute hypoxia produces an increase in ventilation. When the hypoxia is sustained, the initial increase in ventilation is followed a decrease in ventilation. The precise mechanism of this decline in ventilation during sustained hypoxia is unknown. Recent studies hypothesized that the accumulation of dopamine in the central nervous system might have a major role in production of hypoxic respiratory depression. The purpose of this study was to examine whether dopamine has an effect on occurance of central ventilatory depression which is seen in acute hypoxia in peripheral chemoreceptors denervated animals. The experiment were conducted in rabbits anesthetized with Na-pentobarbital (25 mg x kg(-1) i.v.). For intracerebroventricular (i.c.v.) injections of dopamine (1 microg) in each animal, canula was placed in left lateral cerebral ventricle by stereotaxic method. Respiratory frequency (f x min(-1)), tidal volume (V(T)) ventilation minute volume (V(E)) and systemic arterial blood pressure (BP) were recorded during air and 3 minutes hypoxic gas mixture (8%O2-92%N2) breathing. I.c.v. administration of dopamine during normoxia decreased V(T), f, V(E) and BP, significantly. When rabbits were injected with an i.c.v. dopamine on hypoxic gas mixture breathing in control animals, there was depression of hypoxic ventilatory responses. After i.c.v. administration of dopamine antagonists haloperidol (0.1 mg) and domperidone (0.07 mg) in chemodenervated rabbits, the significant increases in V(T), V(E) and BP were observed. On the breathing of hypoxic gas mixture of chemodenervated and i.c.v. dopamine antagonists administrated rabbits, hypoxic depression was completely abolished. These results of this study show that accumulation of dopamine in the brain seems to reduce the response of the central control mechanisms to chemoreceptor impulses during normoxia and hypoxia. In conclusion present study suggests important role played by central dopaminergic pathways in the occurence of acute hypoxic ventilatory depression. PMID- 12086151 TI - Correlation between the numbers of gammadelta T cells and CD4+ HLA-DR+ T cells in broncho-alveolar lavage fluid from patients with diffuse lung disease. AB - CD4+ HLA-DR+ T cells are known to be increasing in broncho-alveolar lavage fluid (BALF) from patients with sarcoidosis, and related to disease activity. Although there are several reports that the number of gammadelta T cells in peripheral blood from patients with sarcoidosis are increasing, contradictory assertions can be seen about the number of gammadelta T cells in BALF, and the clinical significance on the presence of gammadelta T cells in disease site of patients with diffuse lung disease including sarcoidosis. The absolute number of gammadelta T cells and CD4+ HLA-DR+ T cells in BALF were determined by flow cytometry in 107 patients with diffuse lung diseases; 56 with sarcoidosis, 36 with collagen vascular diseases with lung involvement and 15 with idiopathic pulmonary fibrosis. We also measured the number of the transferrin receptor positive macrophages in BALF. The correlation between gammadelta T cells and activated (maybe antigen-specific) T cells and macrophages were evaluated. Sarcoidosis patients were also evaluated from the data of the number of gammadelta T cells in peripheral blood by flow cytometry and clinical backgrounds. A significant correlation between the numbers of these two cell types was detected in each of the three patient groups. The percentage of peripheral gammadelta T cells was markedly increased in 7 sarcoidosis patients, each of whom also showed affected organs other than lung, however, 5 individuals did not show an increased number of gammadelta T cells in BALF. The number of gammadelta T cells in BALF did not correlate with the number of transferrin receptor-positive macrophages in all three patient groups. These results suggest that the increased number of gammadelta T cells in diffuse lung diseases likely plays a role in immunosurveillance and contributes to the activation of antigen specific alphabeta T cell. PMID- 12086152 TI - Congenital central hypoventilation syndrome: a novel mutation of the RET gene in an isolated case. AB - Recently, a few genetic abnormalities were identified in congenital central hypoventilation syndrome (CCHS or Ondine's curse). CCHS is often associated with other neurocristopathies, especially with Hirschsprung's disease (HSCR). Mutations of the genes involved in the receptor tyrosine kinase RET (REarranged during Transfection) (RET)-glial cell line-derived neurotrophic factor (GDNF) and/or endothelin 3 (EDN3)-endothelin receptor-B (EDNRB) signaling pathway have been found in some of HSCR patients. In this study, we analyzed candidates for HSCR, namely the RET, GDNF, EDN3 and EDNRB genes in three isolated CCHS patients to confirm the hypothesis that some CCHS patients have a common genetic abnormality with patients having HSCR or other neurocristopathies. We found a novel R114H mutation of the RET gene in one patient. The R114H mutation is unlikely to be a polymorphism and appears to be associated with CCHS. In addition, we also examined the HOX11L2 (RNX) gene, for which knock-out mice showed CCHS-like syndrome in these isolated CCHS patients and did not detected any mutation. Further cases should be analyzed for more candidates to clarify the pathophysiology of CCHS. PMID- 12086149 TI - Papillomavirus E1 proteins: form, function, and features. AB - The E1 proteins are the essential origin recognition proteins for papillomavirus (PV) replication. E1 proteins bind to specific DNA elements in the viral origin of replication and assemble into hexameric helicases with the aid of a second viral protein, E2. The resultant helicase complex initiates origin DNA unwinding to provide the template for subsequent syntheses of progeny DNA. In addition to ATP-dependent helicase activity, E1 proteins interact with and recruit several host cell replication proteins to viral origin, including DNA polymerase alpha and RPA. This review will compare the basic structures and features of the human (HPV) and bovine (BPV1) papillomaviruses with an emphasis on mechanisms of replication function. PMID- 12086153 TI - Comparative immunohistochemical localizations of aquaporin-1 and aquaporin-4 in the cochleae of three different species of rodents. AB - The species-specific difference of the immunohistochemical localization of aquaporin-1 (AQP1) and aquaporin-4 (AQP4) was investigated in the cochleae of the 3 different species of rodents, including guinea pig, mouse and Mongolian gerbil. In the guinea pig cochlea, intense AQP1-like immunoreactivity was present in the type III fibrocytes in the spiral ligament and the mesenchymal cells just below the basilar membrane. Immunostaining was also found in some type IV fibrocytes in the spiral ligament, fibrocytes in the spiral limbus and mesenchymal cells lining the perilymphatic space against the bony otic capsule. In contrast, no remarkable immunostaining was found in the basilar membrane of the mouse cochlea. The medial part of the Reissner's membrane was positively immunostained with anti-AQP1 antibody only in the mouse cochlea. In the gerbil cochlea, AQP1-like immunoreactivity was weak compared with the other 2 species. AQP4 was found in the cochlear supporting cells, including Claudius cells, Hensen's cells and inner sulcus cells of the 3 rodent species. AQP4 was also expressed in some interdental cells of the spiral limbus. Weak immunoreactivity was also found in the root cells only in the upper turns of the guinea pig cochlea. In contrast, no detectable immunoreactivity was found in the root cells of the other 2 species. The results obtained in the present study provide the first evidence for the existence of the species differences in the expression of the AQP1 and AQP4 proteins in the rodent cochlea. PMID- 12086154 TI - Hypoxia enhances the expression of plasminogen activator inhibitor-1 in human lung cancer cells, EBC-1. AB - Plasminogen activator inhibitor-1 (PAI-1) is one of the target genes of hypoxia inducible factor-1alpha (HIF-1alpha). Besides being an important physiological regulator of the fibrinolytic system PAI-1 is also involved in cancer invasiveness. HIF-1alpha is expressed in various types of pulmonary cells, but the relation of PAI-1 to HIF-1alpha under hypoxic condition in these cells are not fully elucidated. We, therefore, studied the effect of hypoxia on the expression of PAI-1 in a lung cancer cell line EBC-1. The expression of HIF 1alpha protein in EBC-1 cells was enhanced by hypoxia, and this was associated with increased secretion of PAI-1. Hypoxia did not affect the levels of HIF 1alpha mRNA but enhanced the PAI-1 mRNA. Pretreatment of the cells with MG132, which inhibits the proteasomal degradation of HIF-1alpha, increased the production of PAI-1 under both normoxia and hypoxia. We conclude that hypoxia induces PAI-1 expression, in EBC-1 cells, through the stabilization of HIF-1 complex and this may be related to cancer metastasis. PMID- 12086155 TI - Effects of interleukin-18 on diaphragm muscle contraction on rats. AB - To evaluate the role of interleukin (IL)-18 in endotoxin-induced diaphragm muscle deterioration, we studied three treatment groups, namely, a saline+endotoxin group, an IL-18+endotoxin group and an IL-18 only group using Wistar rats. Five minutes after saline or IL-18 (0.25 microg) injection, E. coli endotoxin (30 mg/kg) was injected intraperitoneally. In the saline+endotoxin group, the force frequency curves by ANOVA (p < 0.01), twitch tension (TT) and slope during contraction time (TT/CT) were significantly lower at 4 hours than those at 0 hours due to endotoxin (p < 0.01 and p < 0.05, respectively) and nitric oxide (NO) production had increased at 4 hours as shown by NADPH diaphorase staining. In the IL-18+endotoxin group, the decrement of the force-frequency curves in a higher frequency range (50-120 Hz), and the decrements of TT and TT/CT observed at 4 hours in the saline+endotoxin group were significantly prevented, and NO production was blocked at 4 hours. In the IL-18 only group, the force-frequency curves did not change except for fatigability, and NO production did not occur. It is suggested that IL-18 itself naturally blocks NO production, even when endotoxin coexists, resulting in the prevention of deterioration of diaphragm muscle contraction by endotoxin. PMID- 12086156 TI - Circadian changes in urinary bicarbonate, nitric oxide metabolites and pH in female player during handball camp involved in an exercise, rest and sleep cycle. AB - Bicarbonate and nitric oxide levels are important humoral factors in the blood and are affected by the human body's physical condition. There are few reports, however, on changes in blood bicarbonate and nitric oxide levels during exercise and rest. Since urinary bicarbonate and nitric oxide metabolites reflect the levels of bicarbonate and nitric oxide in the blood, we studied circadian changes in 6 female athletes by monitoring their urinary pH and their levels of urinary bicarbonate and nitric oxide metabolites. Measurements were taken during exercise, rest and sleep. Six female athletes participated in a 3-day team handball training camp where they followed a schedule of exercise, rest and sleep. Urinary samples were collected immediately before and after handball training, at bed-time and upon waking. The urinary pH and levels of urinary bicarbonate and nitric oxide metabolites, including nitrite and nitrate, were examined with a blood gas analyzer and a NOx analyzer. The samples collected after handball training, as compared to the samples taken before exercise, showed a decreased pH, a decrease in levels of bicarbonate and little change in NO metabolites. During rest, urinary bicarbonate, NO metabolites and pH increased markedly in all 6 subjects. The levels of urinary bicarbonate, NO metabolites and pH significantly decreased upon waking. This study took into account the subjects' various physiological conditions when considering the significance of their changes in urinary bicarbonate, NO metabolites and pH during the 3 day handball training program. There were significant circadian changes in the urinary pH, and in the levels of urinary bicarbonate and nitric oxide metabolites, in the athletes involved in the exercise, rest and sleep program at team handball camp. PMID- 12086157 TI - Subsequent progression to membranous glomerulonephritis following exacerbation of urticarial rash in systemic lupus erythematosus: report of 2 cases. AB - Two Japanese female adolescents with systemic lupus erythematosus (SLE), known cases of urinary tract involvements: one with biopsy-proven class II lupus nephritis and the other one with lupus cystitis without overt glomerulonephritis (silent lupus), who after more than 4 years' observation presented with subsequent progression to membranous glomerulonephritis (MGN) following exacerbation of urticarial rash. Although it is well known that lupus nephritis shows histological transformation with time, the late progression to MGN from another World Health Organization histologic pattern has been reported to be less common in pediatric-onset SLE. Although pathogenesis of their MGN remains speculative, these clinical observation might suggest that a possible association between exacerbation of urticarial rash and subsequent progression to MGN in the selected patients with SLE. PMID- 12086158 TI - Evaluation of a recombinant antigen enzyme-linked immunosorbent assay (ELISA) in the diagnostics of antinuclear antibodies (ANA) in children with rheumatic disorders. PMID- 12086159 TI - Familial occurrence of autoimmune diseases and autoantibodies in a Caucasian population of patients with systemic lupus erythematosus. AB - To determine the prevalence of autoimmune diseases and autoantibodies in relatives of Caucasian patients with systemic lupus erythematosus (SLE) we questioned 118 patients for the prevalence of autoimmune diseases in their relatives. Multicase SLE families were selected for further investigation: assessment of the presence of antinuclear antibodies (ANA), thyroid antibodies and IgM rheumatoid factor (IgM-RF). Thirty-three patients reported the presence of 50 autoimmune diseases in 43 relatives. Twenty-two diagnoses could be either confirmed (n=14) or refuted (n=8). SLE clustered significantly within families of SLE patients. Multiple sclerosis and rheumatoid arthritis also seemed to cluster within families of lupus patients. The prevalence of ANA (24%) and thyroid antibodies (44%) in 29 relatives of multicase SLE families was raised (P<0.05). In conclusion, the prevalence of autoimmune diseases is raised in relatives of Caucasian SLE patients. Also, the prevalence of autoantibodies is raised in relatives of multicase SLE families, both suggesting genetic influences in the pathogenesis of the disease. These findings support the genome-wide screening of SLE patients to unravel factors responsible for genetic susceptibility to SLE. PMID- 12086160 TI - Exposure to solvents in female patients with scleroderma. AB - The role of exposure to solvents was investigated in female patients with connective tissue disease and Raynaud's phenomenon using a questionnaire. Sixteen out of the 63 patients with systemic sclerosis had been exposed to solvents. A borderline significance was demonstrated compared to matched female controls (P < 0.05). Fourteen out of the 66 patients with undifferentiated connective tissue disease, 18/86 of patients with Raynaud's phenomenon, 6/45 with systemic lupus erythematosus, 1/16 with dermatopolymyositis, 1/15 with rheumatoid arthritis and 0/13 with primary Sjogren's syndrome had been exposed to solvents. None of these groups of patients showed a statistical significance compared to matched controls. Our present findings indicate that, at least in certain areas of the world, exposure to solvents may be a provoking factor in female scleroderma, but it must be emphasised that only a borderline significance was found between the scleroderma patients and controls. A large multicenter study seems to be required to clarify the importance of solvents as provoking factors of scleroderma. Furthermore, exposure to solvents does not seem to be a provoking factor among females for the other connective tissue diseases. PMID- 12086161 TI - Functional long-term changes in patients with spondylarthropathy. AB - The objective of the study was to determine the long-term functional outcome of chronic spondylarthropathy (SpA) when measured by various functional indices. This information is important in the planning of long-term intervention studies and selection of the best follow-up methods. The study group consisted of 65 patients (21 women and 44 men) with SpA. Their mean age was 49 years and the mean age from diagnosis was 12 years. They completed several questionnaires (developed for the evaluation of functional capacity or the state of health of patients with SpA) at the beginning of the study and 3 years later. The questionnaires were: the Bath Ankylosing Spondylitis Functional Index (BASFI), the Dougados Functional Index (DFI), the Health Assessment Questionnaire for Spondylarthropathies (HAQ S), the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), the Bath Ankylosing Spondylitis Patient Global Assessment (BAS-G) and Stiffness-VAS (on a visual analogue scale). We also asked the patients to inform us if something had occurred in their lives during the follow-up that might have affected their health. Most of the indices (BASFI, DFI, BASDAI, BAS-G and Stiffness-VAS, but not HAQ-S) showed a statistically significant reduction in the functional capacity or state of health of the patients during the follow-up. Many factors occurring during the 3-year period may have influenced the results of the indices. The natural course of the functional capacity of patients with SpA appears to be one of impairment, when evaluated using these indices. Our experience also showed that it is very difficult to separate any effect of a single treatment intervention (e.g. rehabilitation) in a long-term follow-up study, as so many interfering factors, e.g. life events and health-related factors, may affect the follow-up population over several years. PMID- 12086162 TI - Comparison of gastroduodenal, renal and abdominal fat biopsies for diagnosing amyloidosis in rheumatoid arthritis. AB - The aim of the study was to determine the frequency of amyloidosis detected by gastroduodenal biopsy in rheumatoid arthritis (RA) patients, and to investigate correlations between the results of gastroduodenal biopsy and abdominal fat and renal biopsies. A total of consecutive 1006 RA patients underwent gastroduodenal biopsy. The 71 patients who tested positive for gastrointestinal (GI) amyloidosis were asked to undergo renal and abdominal fat biopsies, and 21 did so. Renal biopsies were also performed on 12 patients with no amyloidosis but indicators of drug-induced renal damage, and abdominal fat biopsies were performed on 50 RA patients with no indication of amyloidosis. The prevalence of GI amyloidosis was 7.1%. Urinary abnormalities and GI symptoms were common in GI amyloidisis, and inflammatory markers were elevated. Sixty-one (86%) had either depressed creatinine clearance or urinary symptoms. Nineteen of the 21 patients (91%) with GI amyloidosis who underwent renal biopsies also had renal amyloid deposits. Eleven of the 21 (52%) had amyloidosis on abdominal fat biopsy. None of the 12 patients without GI amyloidosis had renal amyloidosis on renal biopsy, and none of the 50 patients without GI amyloidosis had amyloidosis on abdominal fat biopsy. Gastroduodenal biopsy reveals a high prevalence of amyloidosis in RA patients. Amyloidosis is often associated with signs of renal impairment. Results of GI biopsy are highly correlated with those of renal biopsy, but the results of fat biopsy are not. We recommend GI biopsy for RA patients for the screening of systemic amyloidosis. PMID- 12086163 TI - Echocardiographic evidence of cardiac involvement in ankylosing spondylitis. AB - Aortic insufficiency, myocardial fibrosis and conduction disturbances are known complications of ankylosing spondylitis (AS). However, few studies have assessed left ventricular diastolic function and no data are available about P-wave analysis. In this study 88 AS patients and 31 healthy volunteers underwent clinical examination, electrocardiography, echocardiography and signal-averaged P wave analysis for the evaluation of asymptomatic cardiac involvement. The aortic root in AS patients was larger and this was correlated with the duration of the disease. Five of 88 AS patients (5.7%) had evidence of mitral valve prolapse, six (6.8%) had thick and redundant mitral valves without prolapse, five (5.7%) had mild mitral regurgitation, two had moderate (2.3%) and two had mild (2.3%) aortic regurgitation. Examination of diastolic function revealed a lower peak of E-wave velocity (E) and E/A ratio, a higher peak of A-wave velocity (A) and acceleration rate of the A wave, a longer deceleration time of E-wave velocity and isovolumic relaxation time in the AS group compared to controls. Mean filtered P-wave duration (PWD) in AS was similar to that of controls. However, PWD in AS patients was positively correlated with left atrial dimension and acceleration rate of the A wave and negatively correlated with E and E/A ratio. In conclusion, cardiac involvement may be seen in AS patients in the absence of clinical manifestations. Echocardiographic examination of diastolic function can be used in this asymptomatic period. Further studies are needed to clarify the prognostic significance of diastolic abnormalities and the value of P-wave analysis in cardiac evaluation of these patients. PMID- 12086164 TI - Bone mineral density in women with systemic lupus erythematosus. AB - The aim of this cross-sectional study was to determine the prevalence of reduced bone mineral density (BMD) in a group of female SLE patients and to identify factors predictive of reduced BMD. Femoral neck (FN) and lumbar spine (LS) dual energy X-ray absorptiometry results were evaluated in 79 pre- and postmenopausal women with SLE aged (mean, range) 49 (22-73) years). Variables evaluated were disease duration, SLEDAI, current and cumulative corticosteroid dose, Steinbrocker's functional classification, use of immunosuppressive agents, and history of fracture due to minor trauma. A T-score of < or = 1.0 was found in 61.9% at the LS and 48.3% at the FN, and 18 (23.7%) patients belonged to the category of osteoporosis at LS, compared to only three (5.4%) patients at FN. A statistical difference (P = 0.014) was found when comparing LS BMD in pre- and postmenopausal patients. LS BMD had a significant correlation with daily and cumulative steroid dose (P = 0.016 and 0.031, respectively). There was a significant difference in LS BMD between the daily steroid dose group receiving < or = 7.5 and those receiving > 7.5 mg/day (P = 0.008), and also in FN BMD comparing groups on 0 and > 7.5 mg/day (P = 0.022). There was significant difference in LS and FN BMD between patients in Steinbrocker classes I and III (P = 0.016 and 0.005, respectively). No significant correlation was found in either subgroup between BMD and other studied parameters. We concluded that the prevalence of reduced bone mass at LS is pronounced among postmenopausal women with SLE, in those with a high Steinbrocker functional classification and those on a high daily steroid dose. Therefore, these patients should be considered as a high-risk group deserving regular spinal BMD scans and therapy in time to prevent vertebral fractures. PMID- 12086165 TI - Prevalence of radiographic knee osteoarthritis in Saudi Arabia. AB - In this cross-sectional study we looked at the radiographic evidence of osteoarthritis (OA) of the knee in 300 randomly chosen patients attending 14 primary care facilities for different medical conditions. Radiographic OA was seen in 89 out of 167 males (53.3%) and in 81 out of 133 females (60.9%). The patella was involved with radiographic osteoarthritic changes in 80.7% and 87.8% of female and male OA patients, respectively. In most cases the medial compartment joint space was narrowed. Radiographic OA of the knee is very common in Saudi patients, especially in the patellofemoral compartment. PMID- 12086167 TI - Cross-sectional and longitudinal study of osteoporosis in patients with rheumatoid arthritis. AB - To elucidate the pathology of osteoporosis associated with rheumatoid arthritis (RA), bone mass measurements were performed in 146 female patients with RA and compared with those in 150 age-matched female patients with osteoarthritis (OA) and postmenopausal osteoporosis (OP). Bone mineral density (BMD) was measured at the lumbar spine (L-BMD), the mid-radius (MR-BMD) and the calcaneus (C-BMD) by dual-energy X-ray absorptiometry (DXA), and at the distal radius by peripheral quantitative computed tomography (pQCT). The RA group showed significantly lower BMD at all sites, except L-BMD, than the OA group. Compared with the OP group, the RA group showed a significantly higher L-BMD but no difference at other sites. BMD in RA decreased with disease severity at all sites and lean body mass was highly correlated with L-BMD and C-BMD. Cross-sectional analysis revealed early bone loss at the distal radius and a decrease of L-BMD, MR-BMD, and C-BMD with disease duration. Longitudinal analysis showed that the annual loss of L BMD, MR-BMD and C-BMD tended to be lower with increasing disease duration. Glucocorticoid administration had no influence on L-BMD, MR-BMD or C-BMD. We concluded that, unlike postmenopausal osteoporosis, osteoporosis associated with RA is characterised by relatively preserved bone mass in the axial bone and marked loss in the peripheral bone. The risk factors for generalised osteoporosis are a long disease duration, severity of disease, and decreased lean body mass. PMID- 12086166 TI - Remitting seronegative symmetrical synovitis with pitting oedema: a study of 12 cases. AB - Twelve patients with remitting seronegative symmetrical synovitis with pitting oedema (RS3PE) were analysed. Eight of them had typical RS3PE without underlying disease, and four presented associated neoplasia. The first patients experienced an excellent response to low doses of prednisone, and they all achieved complete and permanent remission. The mean treatment duration was 18 months and the mean follow-up was 4.4 years. During the follow-up, none of these patients relapsed, had fever or general health deterioration, and hand and foot radiographs did not show erosion. One of them developed a panarteritis nodosa 6 years later. Four RS3PE patients had associated neoplasia. Two were with solid malignancies, and the other two presented haematological malignancies. In one of them RS3PE preceded the diagnosis of malignancy. The diagnosis of RS3PE in the other patients was subsequent to cancer. The first patients presented clinical characteristics suggestive of paraneoplastic RS3PE, and they had a poor response to corticosteroid therapy. Two patients died, and the rest of them had a complete response to surgical resection of the tumour or to chemotherapy. In general, idiopathic RS3PE patients do not show either general health deterioration or fever and they do respond to low doses of steroids (10 mg/day). We observed strong contrasts with the results obtained when treating RS3PE patients with associated neoplasia. In patients with RS3PE the presence of systemic symptoms along with resistance to low doses of corticosteroid therapy should alert the physician to the possible presence of malignancy. PMID- 12086168 TI - An assessment of histopathological criteria for infection in joint arthroplasty in rheumatoid synovium. AB - Intraoperative frozen section is reported to be a reliable means of identifying occult infection for preoperative evaluation of arthroplasty. The aim of this study was to determine whether the reported histopathological criteria--the existence of more than 10 polymorphonuclear cells (PMN) per high-power field--is valuable for determination of infection during the arthroplasty of patients with rheumatoid arthritis (RA). The permanent histological sections of RA synovium were analysed to study the degree of infiltration of PMNs. Furthermore, in order to examine the penetrative distribution of PMNs within the synovial tissues, immunohistochemical staining of PMNs was performed. In addition, the clinical history, from the postoperative period to the present, was investigated in 46 patients (60 joints). The presence of early- and/or late-stage postoperative infection, the development of postoperative fever, the progression of erythrocyte sedimentation rate (ESR) (more than 30 mm per hour) and the changes in CRP (more than 10 mg per litre) were further examined and compared with the histopathological tissue analyses and findings. The results demonstrated the presence of more than five PMNs per high-power field, excluding surface fibrin and inflammatory exudate in at least five separate microscopic fields in 10 joints (16.7%) of nine patients, out of 60 joints of 46 patients, in which no postoperative infection was evident. As to the magnitude of penetrative distribution of PMNs in 10 joints, there was a trend of deepening infiltration among the patients with intensive PMN infiltration. In addition, no development of postoperative fever, CRP or continuous indications of high ESR values were evident in this group. As the existence of more than 10 PMN per high-power field was not absolutely indicative of occult infection, investigation of frozen section during arthroplasty should be carefully managed. PMID- 12086169 TI - Scleroderma renal crisis and concurrent isolated pulmonary hypertension in mixed connective tissue disease and overlap syndrome: report of two cases. AB - We here report on scleroderma renal crisis (SRC) appearing concurrently with isolated pulmonary hypertension (IPHT), that is, pulmonary hypertension without interstitial lung disease with fibrosis, in a patient with mixed connective tissue disease (MCTD) and in one with overlap syndrome or undifferentiated connective tissue disease (UCTD). To the best of our knowledge there are only five previous reports on SRC in MCTD and UCTD. The unexpected appearance of SRC in the setting of concomitant IPHT and limited or absent scleroderma skin changes is discussed. PMID- 12086170 TI - Ochronosis: a case of severe ochronotic arthropathy. AB - Ochronosis involves primarily the large cartilaginous joint surfaces, ribs, intervertebral discs, ear cartilage etc. We report on a 53-year-old woman with typical alkaptonuric ochronosis with dark urine, blue-black pigmentation of the auriculae and hands, focal brown hyperpigmentation of the sclera, spondylarthropathy and severe shoulder joint involvement. PMID- 12086171 TI - Polyarticular heterotopic ossification complicating drug-induced coma. AB - The authors describe a case of generalised heterotopic ossification (HO) complicating the clinical course of a 31-year-old male patient with a history of Hodgkin's lymphoma and of long stay in an intensive care unit, where he had been placed in an artificial coma. The clinical features, as well as radiological findings and therapy, are discussed. PMID- 12086172 TI - Behcet's disease with severe arterial involvement in a child. AB - Behcet's disease is a vasculitis characterised by its thrombotic tendency. In some patients, manifestations of vascular lesions may dominate the clinical picture instead of the classic triad. We describe a 13-year-old boy with a 1.5 year history of Behcet's disease presenting with complaints of acute abdominal pain, severe headache and decreased vision. His work-up revealed a saccular aneurysm of the distal abdominal aorta, thrombosis in the right common iliac, external iliac and femoral arteries, and thrombosis of the superior sagittal sinus. Aortoiliac bypass with a Dacron graft was performed with success. He also received cyclosporin A and anticoagulant therapy. Severe vasculitis may become overt at any age in patients with Behcet's disease. Early diagnosis and management is important to prevent morbidity and mortality. PMID- 12086173 TI - The problem of empyematous pleural effusion in rheumatoid arthritis: report of two cases and review of the literature. AB - Two patients with rheumatoid arthritis and empyematous pleural effusion were treated with repeated drainage and intrapleural corticosteroids. One patient with active joint disease improved within 3 months without sequelae, probably because of the systemic therapy. The other patient, with non-active joint disease, had persistent pleural effusion which resulted in pleural thickening and symptomatic restrictive disturbance. It appears that early intervention intended to prevent the accumulation of empyematous pleural effusion could also prevent pleural thickening and fibrosis. Therapeutic options are discussed. PMID- 12086175 TI - Can outcome of early synovitis be predicted by serological tests? PMID- 12086174 TI - Occult liver involvement by polyarteritis nodosa. AB - The diagnosis of panarteritis nodosa is usually difficult because of the variability of clinical manifestations. Herein is presented a case of occult liver involvement by polyarteritis nodosa. Several biopsies were performed but the diagnosis was made with the contribution of a hepatic arteriography. PMID- 12086176 TI - Flow cytometric detection and quantification of heterotrophic nanoflagellates in enriched seawater and cultures. AB - A flow cytometric protocol to detect and enumerate heterotrophic nanoflagellates (HNF) in enriched waters is reported. At present, the cytometric protocols that allow accurate quantification of bacterioplankton cannot be used to quantify protozoa for the following reasons: i) the background produced by the bacterial acquisitions does not allow the discrimination of protozoa at low abundance, ii) since the final protozoan fluorescence is much higher than the bacterioplankton fluorescence (more than 35 fold) the protozoa acquisitions lie outside the range. With an increase in the fluorescence threshold and a reduction of the fluorescence detector voltage, low fluorescence particles (bacteria) are beneath the detection limits and only higher fluorescence particles (most of them heterotrophic nanoflagellates) are detected. The main limitation for the application of the cytometric protocol developed is that a ratio of bacteria/HNF below 1000 is needed. At higher ratios, the background of larger cells of bacterioplankton makes it difficult to discriminate protozoa. The proposed protocol has been validated by epifluorescence microscopy analyzing both a mixed community and two single species of HFN: Rhynchomonas nasuta and Jakoba libera. Taking into account the required bacteria/HNF ratio cited above, the results provide evidence that the flow cytometric protocol reported here is valid for counting mixed communities of HNF in enriched seawater and in experimental micro or mesocosms. In the case of single species of HNF previous knowledge of the biological characteristics of the protist and how they can affect the effectiveness of the flow cytometric count is necessary. PMID- 12086177 TI - On the specificity of PCR detection of Listeria monocytogenes in food: a comparison of published primers. AB - A total of nine pairs of primers, seven previously published and two newly developed, have been assayed for PCR detection of Listeria monocytogenes in food. They have been tested for specificity on a total of 72 strains including reference and food isolates belonging to L. monocytogenes and other species in the genus. First of all, a polyphasic approach has been carried out in order to establish a reference strain collection. They were biochemically and genetically characterized by API-Lis and randomly amplified polymorphic DNA PCR (RAPD-PCR), respectively. Random amplification of DNA was performed with M13, T7 and T3 universal primers and a data bank was created to compile the RAPD patterns of all the analyzed strains. The UPGMA cluster analysis of RAPD profiles with primer M13 showed eight clusters at 72.3% similarity. Clusters 2 and 7 corresponded to L. monocytogenes. Clusters 1 and 6 grouped L. ivanovii strains. Clusters 3, 4, 5 and 8 corresponded to L. grayi, L. innocua, L. welshimeri and L. seeligeri, respectively. Pattern analysis revealed the existence of miss-identified reference strains which was confirmed by 16S rDNA sequence analysis. RAPD-PCR is a rapid genetic test which helped to confirm strain identity. On the basis of PCR specificity results, primers LM1-LM2 were the best combination for the detection of L. monocytogenes since they only amplified the specific fragment in strains that had been genetically and biochemically assessed as belonging to the species. Specificity of other assayed primers is discussed. PMID- 12086178 TI - Characterization of Carnobacterium divergens strain 6251 isolated from intestine of Arctic charr (Salvelinus alpinus L.). AB - An atypical strain of Carnobacterium divergens, strain 6251, was isolated from the small intestine of Arctic charr (Salvelinus alpinus L.), fed high dietary carbohydrate. This strain showed marked growth inhibitory effects in vitro against the fish pathogens Aeromonas salmonicida subsp. salmonicida (furunculosis), Vibrio anguillarum (vibriosis) and Vibrio viscocus (winter ulcer). The strain is a non-motile Gram-positive psychrotrophic rod that lacks both catalase and oxidase, grows at pH 9.1 (CTAS agar), but not on acetate containing media (pH < or = 5.4), on TCBS or at < or =6% sodium chloride content. Strain 6251 is facultatively anaerobic and utilises tryptone as a sole source of nutrient. Further characterisation showed the most abundant cellular fatty acid of strain 6251 to be oleic acid (18:1) (n-9) (36.0%). Sequencing of a 16S rDNA region of 578 nucleotides and AFLP microbial fingerprinting suggested that strain 6251 is not closely related to any carnobacteria known, however, DNA-DNA similarity determinations showed high similarity (96.2%) with the type strain of Carnobacterium divergens. The unique phenotypic attributes of this strain represent new information on the biodiversity and ecology of carnobacteria and especially of the species C. divergens. PMID- 12086179 TI - Purification and characterisation of an extracellular fructan beta-fructosidase from a Lactobacillus pentosus strain isolated from fermented fish. AB - Lactobacillus pentosus B235, which was isolated as part of the dominant microflora from a garlic containing fermented fish product, was grown in a chemically defined medium with inulin as the sole carbohydrate source. An extracellular fructan beta-fructosidase was purified to homogeneity from the bacterial supernatant by ultrafiltration, anion exchange chromatography and hydrophobic interaction chromatography. The molecular weight of the enzyme was estimated to be approximately 126 kDa by gel filtration and by SDS-PAGE. The purified enzyme had the highest activity for levan (a beta(2-->6)-linked fructan), but also hydrolysed garlic extract, (a beta(2-->1)-linked fructan with beta(2-->6)-linked fructosyl sidechains), 1,1,1-kestose, 1,1-kestose, 1-kestose, inulin (beta(2-->1)-linked fructans) and sucrose at 60, 45, 39, 12, 9 and 3%, respectively, of the activity observed for levan. Melezitose, raffinose and stachyose were not hydrolysed by the enzyme. The fructan beta-fructosidase was inhibited by p-chloromercuribenzoate, EDTA, Fe2+, Cu2+, Zn2+ and Co2+, whereas Mn2+ and Cu2+ had no effect. The sequence of the first 20 N-terminal amino acids was: Ala-Thr-Ser-Ala-Ser-Ser-Ser-Gln-Ile-Ser-Gln-Asn-Asn-Thr-Gln-Thr-Ser-Asp-Val Val. The enzyme had temperature and pH optima at 25 degrees C and 5.5, respectively. At concentrations of up to 12% NaCl no adverse effect on the enzyme activity was observed. PMID- 12086180 TI - Symbiotic and taxonomic diversity of rhizobia isolated from Acacia tortilis subsp. raddiana in Africa. AB - A collection of rhizobia isolated from Acacia tortilis subsp. raddiana from various sites in the North and South of Sahara was analyzed for their diversity at both taxonomic and symbiotic levels. On the basis of whole cell protein (SDS PAGE) and 16S rDNA sequence analysis, most of the strains were found to belong to the Sinorhizobium and Mesorhizobium genera where they may represent several different genospecies. Despite their chromosomal diversity, most A. tortilis Mesorhizobium and Sinorhizobium symbionts exhibited very similar symbiotic characters. Nodulation tests showed that the strains belong to the Acacia Leucaena-Prosopis nodulation group, although mainly forming non-fixing nodules on species other than A. tortilis. Most of the strains tested responded similarly to flavonoid nod gene inducers, as estimated by using heterologous nodA-lacZ fusions. Thin layer chromatography analysis of the Nod factors synthesized by overproducing strains showed that most of the strains exhibited similar profiles. The structures of Nod factors produced by four different Sinorhizobium sp. strains were determined and found to be similar to other Acacia-Prosopis-Leucaena nodulating rhizobia of the Sinorhizobium-Mesorhizobium-Rhizobium branch. They are chitopentamers, N-methylated and N-acylated by common fatty acids at the terminal non reducing sugar. The molecules can also be 6-O sulfated at the reducing end and carbamoylated at the non reducing end. The phylogenetic analysis of available NodA sequences, including new sequences from A. tortilis strains, confirmed the clustering of the NodA sequences of members of the Acacia-Prosopis-Leucaena nodulation group. PMID- 12086181 TI - Molecular evidence to support the expansion of the hostrange of Chlamydophila pneumoniae to include reptiles as well as humans, horses, koalas and amphibians. AB - The Chlamydiales are a family of unique intracellular pathogens that cause significant disease in humans, birds and a wide range of animal hosts. Of the currently recognized species, Chlamydophila (previously Chlamydia) pneumoniae, unlike the other chlamydial species, has been previously considered to be solely a pathogen of humans, causing significant respiratory disease and has also been strongly connected with cardiovascular disease. Here we report the finding that strains of C. pneumoniae are widespread in the environment, being detected by molecular methods in a range of reptiles (snakes, iguanas, chameleons) and amphibians (frogs, turtles). Of particular interest was the finding that genotyping of the chlamydial major outer membrane protein gene in these newly identified C. pneumoniae strains showed that many were genetically very similar, if not identical to the human respiratory strains. Whether these reptilian and amphibian strains of C. pneumoniae are still capable of infecting humans, or crossed the host barrier some time ago, remains to be determined but may provide further insights into the relationship of this common respiratory infection with its human host. PMID- 12086182 TI - Study of the first hours of microvinification by the use of osmotic stress response genes as probes. AB - When yeast cells are inoculated into grape must for vinification they find stress conditions because of osmolarity, which is due to very high sugar concentration, and pH lower than 4. In this work an analysis of the expression of three osmotic stress induced genes (GPD1, HSP12 and HSP104) under microvinification conditions is shown as a way to probe those stress situations and the regulatory mechanisms that control them. The results indicate that during the first hours of microvinification there is an increase in the GPDI mRNA levels with a maximum about one hour after inoculation, and a decrease in the amount of HSP12 and HSP104 mRNAs, although with differences between them. The RNA steady-state levels of all the genes considered, and in some cases the microvinification progress are significantly affected by the composition of the must (pH, nature of the osmotic agent and carbon source). These results point out the importance of the control of these parameters and the yeast molecular response during the first hours of vinification for an accurate winemaking process. PMID- 12086183 TI - Characterization of Alternaria and Penicillium species from similar substrata based on growth at different temperature, pH and water activity. AB - Fifty-eight Alternaria isolates representing 10 species or species-groups and 66 Penicillium isolates representing 18 species were examined for their growth response to the combined effects of water activity, temperature and pH in an extended Central Composite Design. Growth responses were recorded as colony diameter after one and two weeks of growth and analysed using different multivariate statistical analyses. The isolates, when analysed by Principal Component Analysis, clustered according to their genus and to some degree to species or species groups and not according to substratum as excepted. Soft Independent Modelling of Class Analogy and Response Surface Analysis showed that growth responses or growth profiles may be used as classification tool. Partial Least Squares Regression showed that a combination of two different media based on Dichloran Rose bengal Yeast Extract Sucrose agar incubated at two different temperatures were enough to get genus segregation and to some extent species segregation. The results also showed that water activity, temperature and pH interact strongly in their effect on growth rates and that the squared products (optima) of water activity, temperature and pH for each isolate were important for modelling the data sufficiently. PMID- 12086184 TI - Characterization of a phosphatase secreted by Staphylococcus aureus strain 154, a new member of the bacterial class C family of nonspecific acid phosphatases. AB - An acid phosphatase, designated SapS, hydrolyzing p-nitrophenyl phosphate (pNPP), was identified and characterized from the culture supernatant of a Staphylococcus aureus strain isolated from vegetables. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the protein indicated an estimated molecular mass of 30 kDa. The enzyme displayed optimum activity at 40 degrees C and pH 5. Characterization of the phosphatase in a reconstitution assay showed that MgCl2 and Triton X-100, respectively, restored maximal activity, but not CaCl2 The phosphatase activity was affected by EDTA and sodium molybdate. The DNA sequence encoding SapS was cloned and sequenced. The putative acid phosphatase gene encodes a protein of 296 amino acids with a 31-residue signal peptide. Database searches revealed significant structural homology of SapS to several proteins belonging to the bacterial class C family of nonspecific acid phosphatases. Comparison of the sequences indicated that despite a low level of overall conservation between the proteins, four conserved sequence motifs could be identified. PMID- 12086185 TI - Primary structure of selected archaeal mesophilic and extremely thermophilic outer surface layer proteins. AB - The archaea are recognized as a separate third domain of life together with the bacteria and eucarya. The archaea include the methanogens, extreme halophiles, thermoplasmas, sulfate reducers and sulfur metabolizing thermophiles, which thrive in different habitats such as anaerobic niches, salt lakes, and marine hydrothermals systems and continental solfataras. Many of these habitats represent extreme environments in respect to temperature, osmotic pressure and pH values and remind on the conditions of the early earth. The cell envelope structures were one of the first biochemical characteristics of archaea studied in detail. The most common archaeal cell envelope is composed of a single crystalline protein or glycoprotein surface layer (S-layer), which is associated with the outside of the cytoplasmic membrane. The S-layers are directly exposed to the extreme environment and can not be stabilized by cellular components. Therefore, from comparative studies of mesophilic and extremely thermophilic S layer proteins hints can be obtained about the molecular mechanisms of protein stabilization at high temperatures. First crystallization experiments of surface layer proteins under microgravity conditions were successful. Here, we report on the biochemical features of selected mesophilic and extremely archaeal S-layer (glyco-) proteins. PMID- 12086186 TI - Psychrobacter faecalis sp. nov., a new species from a bioaerosol originating from pigeon faeces. AB - The taxonomy of strain Iso-46T isolated from a bioaerosol generated by cleaning of a pigeon faeces contaminated room was investigated in a polyphasic approach. The beige pigmented Gram-negative, oxidase-negative organism contained a quinone system with mainly ubiquinone Q-8, and the polar lipid profile was composed of phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol, beside some hitherto uncharacterized phospholipids. Major polyamines were spermidine and putrescine and also small amounts of cadaverine. The analysis of the fatty acids revealed 3-OH 12:0 and 3-OH 14:0 (within summed feature 3) as hydroxylated fatty acids. These chemotaxonomic characteristics suggest that the strain belongs to the gamma-subclass of the Proteobacteria namely into the genus Psychrobacter. Analysis of the 16S rRNA gene supported the allocation into the genus Psychrobacter, but showing similarities to all described species of this genus lower than 97%. Iso-46T was able to grow on MacConkey agar and other high nutrient containing media within a temperature range of 4 degrees C to 36 degrees C. On the basis of nutritional and further physiological features, a clear differentiation from all other Psychrobacter species was possible. For these reasons it is proposed to create a new species with the name Psychrobacter faecalis sp. nov. PMID- 12086187 TI - Novosphingobium hassiacum sp. nov., a new species isolated from an aerated sewage pond. AB - The taxonomy of two strains W-51T and W-52 isolated from a wastewater treatment plant was investigated in a polyphasic approach. The yellow pigmented gram negative organism contained a quinone system with mainly ubiquinone Q-10, and the polar lipid profile contained a sphingoglycolipid suggesting that both strains belonged to the the alpha-4 subclass of the Proteobacteria. The polar lipid profile consisted furthermore of phosphatidylethanolamine, diphosphatidylglycerol, and phosphatidylcholine and of minor amounts of phosphatidylglycerol and phosphatidylmonomethylethanolamine. Sequencing of the 16S rRNA gene supported the allocation into the genus Novosphingobium, together with the type strains of N. subterraneum, N. aromaticivorans, N. stygium, and N. capsulatum, showing similarities of 97.3%, 97.0%, 95.7% and 96.2%, respectively. This allocation was supported by the polyamine profile, which consisted mainly of spermidine. The analysis of the fatty acids revealed 2-OH 13:0, 2-OH 14:0 and 2 OH 15:0, with 2-OH 15:0 as predominant hydroxylated fatty acid. W-51T and W-52 were almost identical with respect to their phenotypic including the majority of the chemotaxonomic properties, identical in their 16S rRNA sequences, and showed 86% DNA-DNA similarity. Both strains were able to reduce nitrate and on the basis of further physiological features, a clear differentiation from all other Novosphingobium species was possible. The DNA-DNA similarities of W-51T to the type strains of N. subterraneum, N. aromaticivorans, and N. capsulatum were below 56%. For these reasons, it is proposed to create a new species with the name Novosphingobium hassiacum sp. nov. PMID- 12086188 TI - Anaerostipes caccae gen. nov., sp. nov., a new saccharolytic, acetate-utilising, butyrate-producing bacterium from human faeces. AB - Two strains of a previously undescribed Eubacterium-like bacterium were isolated from human faeces. The strains are Gram-variable, obligately anaerobic, catalase negative, asporogenous rod-shaped cells which produced acetate, butyrate and lactate as the end products of glucose metabolism. The two isolates displayed 99.9% 16S rRNA gene sequence similarity to each other and treeing analysis demonstrated the faecal isolates are far removed from Eubacterium sensu stricto and that they represent a new subline within the Clostridium coccoides group of organisms. Based on phenotypic and phylogenetic criteria, it is proposed that the two strains from faeces be classified as a new genus and species, Anaerostipes caccae. The type strain of Anaerostipes caccae is NCIMB 13811T (= DSM 14662T). PMID- 12086189 TI - Study of the intra- and interlaboratory reproducibility of partial single base C sequencing of the 16S rRNA gene and its applicability for the identification of members of the genus Streptococcus. AB - The use of Single Base C-Sequencing of the first 500 bases of the 16S rRNA-gene (SBCS) combined with capillary electrophoresis was evaluated for the identification of reference strains of 30 different species within the genus Streptococcus. For SBCS, only dd-CTP's are used in the sequencing reactions instead of the four dideoxy bases and the primer is fluorescently labeled. The reproducibility, interlaboratory exchangeability and discriminative power of this method were studied by comparing the patterns obtained in three laboratories under highly standardized conditions. The interlaboratory reproducibility proved to be high, enabling the construction of a common database for the identification of strains belonging to the streptococcal species studied. Most of the examined species generated distinguishable profiles. SBCS did not differentiate between the closely related species S. constellatus and S. intermedius. Also S. thermophilus and S. vestibularis as well as S. mitis and S. pneumoniae showed highly resembling profiles. The previously reported heterogeneity within the species S. equinus was reflected by SBCS. For all other species, strains belonging to the same species generated indistinguishable patterns. In conclusion, Single Base C-sequencing of the first 500 bases of the 16S rRNA-gene could be a useful and widely applicable method for the identification of bacteria at the species level, with the added advantage of being more rapid and easier to automatize than full sequence determination. PMID- 12086190 TI - Differentiation of nocardia species by PCR-randomly amplified polymorphic DNA fingerprinting. AB - Representatives of fifteen validly described and three non-validly described species of Nocardia were assigned to nineteen groups based on an optimised PCR randomly amplified polymorphic DNA fingerprinting technique. Species specific banding patterns were recognised for the representatives of N. brasiliensis, N. crassostreae, N. farcinica, N. otitidiscaviarum and N. seriola. Unique banding patterns were also seen for the type strains of N. brevicatena, N. carnea, N. salmonicida, N. uniformis and N. vaccinii, and for the single representatives of "N. fusca", "N. pseudosporangifera", and "N. violaceofusca". More than one banding pattern was detected for the N. asteroides, N. flavorosea, N. nova, N. pseudobrasiliensis and N. transvalensis strains though in the case of the representative strains of N. nova and N. transvalensis the patterns were similar for each of these species. The results are in line with current trends in nocardial systematics thereby indicating that PCR-randomly amplified polymorphic DNA fingerprinting provides valuable data for the classification and identification of pathogenic nocardiae to the species level. PMID- 12086191 TI - Contrasting nifD and ribosomal gene relationships among Mesorhizobium from Lotus oroboides in northern Mexico. AB - PCR screens for length variation in a 5' portion of 23S ribosomal RNA and in the 3' end of the 16S rRNA-23S rRNA internal transcribed spacer (ITS) region indicated that nodule bacteria from a Mexican population of Lotus oroboides were diverse on a local scale. Three 23S rRNA length variants and five ITS length variants were detected among the 22 isolates. Sequencing of nearly full-length 16S rRNA genes in three isolates indicated that they fell into the genus Mesorhizobium, but comprised two distinct groups. Two isolates were closely related to M. loti LMG 6125T, while the other isolate clustered with an assemblage of Mesorhizobium taxa that included M. amorphae, M. plurifarium and M. huakuii. However, a phylogenetic tree based on 715 bp of the nitrogenase alpha subunit (nifD) gene was significantly discordant with the relationships inferred from rRNA sequences. Two isolates that were nearly identical for 16S rRNA had nifD genes that varied at 2% of sites, and one of these nifD sequences was identical to that of another isolate with a strongly divergent 16S rRNA gene. A plasmid screen followed by Southern hybridization indicated that only one of these strains harbored a plasmid-borne nifD gene. These results imply that gene transfer events have altered the distribution of nifD sequences among lineages within this natural population of Mesorhizobium strains. PMID- 12086192 TI - Molecular characterization of ochratoxin A producing strains of the genus Penicillium. AB - Sixty-six strains classified as P. verrucosum based on morphological criteria were characterized by molecular methods like RAPD, AFLP and ITS sequencing. Two groups could be identified by RAPD and AFLP analyses. The two RAPD as well as the two AFLP groups were completely coincidental. Strains in the two groups differed in their ability to produce ochratoxin A, with group I containing mainly high producing strains, and group II containing moderate to non-producing strains. The strains from group I originate from foods, such as cheeses and meat products, while the strains from group II originate from plants. The ribosomal ITS1-5.8S ITS2 sequences were similar, except for two single nucleotide exchanges in several strains of each group. A chemotaxonomical analysis of some of the strains identified differences between the groups in secondary metabolite production. Strains from group I possessed the chemotype of P. nordicum and strains from group II that of P. verrucosum. The differences at the RAPD and AFLP level, which parallel the chemotypic differences, are consistent with the recent reclassification of ochratoxin A producing penicillia to be either P. verrucosum or P. nordicum. The homolgy between the ITS sequences however indicates phylogenetic relationship between the two species. PMID- 12086193 TI - The microbial community composition of a nitrifying-denitrifying activated sludge from an industrial sewage treatment plant analyzed by the full-cycle rRNA approach. AB - The composition of the microbial community present in the nitrifying-denitrifying activated sludge of an industrial wastewater treatment plant connected to a rendering facility was investigated by the full-cycle rRNA approach. After DNA extraction using three different methods, 94 almost full-length 16S rRNA gene clones were retrieved and analyzed phylogenetically. 59% of the clones were affiliated with the Proteobacteria and clustered with the beta- (29 clones), alpha- (24), and delta-class (2 clones), respectively. 15 clones grouped within the green nonsulfur (GNS) bacteria and 11 clones belonged to the Planctomycetes. The Verrucomicrobia, Acidobacteria, Nitrospira, Bacteroidetes, Firmicutes and Actinobacteria were each represented by one to five clones. Interestingly, the highest 'species richness' [measured as number of operational taxonomic units (OTUs)] was found within the alpha-class of Proteobacteria, followed by the Planctomycetes, the beta-class of Proteobacteria, and the GNS-bacteria. The microbial community composition of the activated sludge was determined quantitatively by using 36 group-, subgroup-, and OTU-specific rRNA-targeted oligonucleotide probes for fluorescence in situ hybridization (FISH), confocal laser scanning microscopy and digital image analysis. 89% of all bacteria detectable by FISH with a bacterial probe set could be assigned to specific divisions. Consistent with the 16S rRNA gene library data, members of the beta class of Proteobacteria dominated the microbial community and represented almost half of the biovolume of all bacteria detectable by FISH. Within the beta-class, 98% of the cells could be identified by the application of genus- or OTU-specific probes demonstrating a high in situ abundance of bacteria related to Zoogloea and Azoarcus sensu lato. Taken together, this study provides the first encompassing, high-resolution insight into the in situ composition of the microbial community present in a full-scale, industrial wastewater treatment plant. PMID- 12086194 TI - Patient, physician, and nurse satisfaction with antibiotics. AB - To prospectively evaluate patient (parent), physician, and nurse satisfaction with antibiotics, 12,102 children in 661 pediatric study centers throughout the United States with infections to be treated on an outpatient basis were followed with questionnaires completed by parents and with interviews of patients and their parents. Comprehensive surveys were also completed by participating pediatricians and their nursing staffs of which 11,913 (98.4%) contained adequate information for inclusion in an analysis. Data were combined and analyzed using standard statistical methods. Loracarbef was perceived as being superior to other antibiotics based on cost, palatability, and dosing intervals. Few differences were seen among most antibiotics based on treatment outcome or adverse events. All antibiotics studied offered acceptable efficacy and were well tolerated as judged by physicians, nurses, parents, and children. Slight differences might result in increased compliance for some patients. PMID- 12086195 TI - The assessment and management of the respiratory complications of pediatric neuromuscular diseases. AB - Respiratory complications are a major cause of morbidity and mortality in the pediatric neuromuscular diseases. Weakness of the muscles of respiration results in shallow breathing and ineffective cough, making patients vulnerable to atelectasis, pneumonia, and tracheal obstruction by retained respiratory tract secretions. Assessment of the risk of respiratory complications includes evaluating the patient's history and respiratory physical examination and measuring pulmonary function and gas exchange. Treatment options include methods of assisted cough and mechanical ventilation. This review emphasizes the use of noninvasive respiratory aids to optimize duration and quality of life in these potentially fatal diseases. PMID- 12086196 TI - Obstructive uropathy: an important cause of chronic renal failure in children. AB - Care of individuals with renal failure continues to impose a tremendous burden on our national health care budget. Chronic renal failure is the final common denominator of a multiplicity of pathologic processes, some of which progress silently over years. Of these, obstructive uropathy is a prominent cause of kidney failure, accounting for 16.5% of all pediatric renal transplants in 1997. The pathophysiology of obstructive uropathy is reviewed as the basis for a differential diagnosis. In view of the significant role played by obstructive uropathy in the development of renal failure, early and definitive treatment of this clinical entity is imperative. PMID- 12086197 TI - Weight and height deficits in children with brain stem tumors. AB - Weight deficit is common among children with brain stem tumors and is often accompanied by height deficit. Among 22 consecutive children (< or =18 years) with brain stem tumor, 16 had weight deficit (< or =20th percentile) (p<1.4e-7). Eleven were at or less than the 5th percentile, and 5 were less than the 1st weight percentile. Eight also had height deficit (< or =20th percentile) (p<0.06). Misdiagnoses occurred: failure to thrive in 5, growth retardation in 2, and anorexia nervosa in 2. Delay between these diagnoses and that of brain tumor averaged 4.5 years. Detailed neuroradiologic study seems worthwhile if weight deficit is extreme and either unexplained or uncorrectable, or if the weight deficit is accompanied by an abnormality suggestive of intracranial disease. PMID- 12086198 TI - Perceptions of physicians and pediatric patients about atopic dermatitis, its impact, and its treatment. AB - The National Eczema Association for Science and Education surveyed 429 patients with atopic dermatitis or their parents and 303 physicians about atopic dermatitis, its effect on quality of life (QOL), and the adequacy of available treatments. The physician survey focused on disease severity and management; the patient survey, on disease severity, impact on QOL, and treatment preferences. Physician and patient/parent respondents generally agreed about their assessments of disease severity, poor effectiveness of over-the-counter products, and the need for more patient support groups and toll-free phone information. Atopic dermatitis significantly impacts QOL in children, particularly in those with moderate-to-severe disease. New treatments should offer enhanced efficacy, less frequent application, and fewer adverse effects. PMID- 12086199 TI - One-year trial on safety and normal linear growth with flunisolide HFA in children with asthma. AB - Flunisolide hydrofluoroalkane (HFA) has efficacy equivalent to that of flunisolide chlorofluorocarbon (CFC) at one third the dose of the CFC formulation, a reduction from 250 microg/puff for flunisolide CFC to 85 microg/puff for flunisolide HFA. Flunisolide HFA delivers a smaller particle size (1.2 microm) in solution, resulting in improved lung deposition as compared with flunisolide CFC (3.8 microm), which is delivered in suspension. An added built-in spacer has reduced oropharyngeal deposition that may result in fewer adverse events and make it easier to use. The objective of this study was to compare the year-long safety of flunisolide HFA (daily dosage 340 microg) with that of CFC beclomethasone dipropionate (BDP) (daily dosage 336 microg) and cromolyn sodium (daily dosage 6,400 microg) in children 4-11 years old with mild-to-moderate asthma. The effects of these drugs on linear growth and growth velocity were also compared. The study was a 1-year open-label, parallel-group trial. Changes in physical examinations (including growth), adverse events, vital signs, electrocardiograms, cosyntropin stimulation tests, mouth and throat cultures for Candida albicans, and laboratory findings were analyzed. Patients 4-5 years old received flunisolide HFA only. In total, 235 children were evaluated (152 receiving flunisolide HFA, 39 BDP, and 44 cromolyn). The incidence of adverse events was comparable among treatment groups; most were mild or moderate and considered unrelated to treatment. Among patients 6-11 years old, mean changes from baseline height at week 52 were 6.2 cm for the flunisolide HFA and cromolyn groups and 5.1 cm for the BDP group. Thus growth in children receiving flunisolide HFA was unaffected by 1 year of treatment. Changes from baseline in other parameters, including response to cosyntropin stimulation, were insignificant and similar among the 3 treatment groups. At the dosages studied, and following 1 year of treatment, flunisolide HFA with its small particle size and built-in spacer is safe and well tolerated in children 4-11 years old. There are no adverse effects associated with hypothalamic pituitary axis (HPA) function of flunisolide HFA, including linear growth in children 6-11 years old when compared with BDP and cromolyn sodium. PMID- 12086200 TI - Smoking cessation counseling: training and practice among women pediatricians. AB - We examined characteristics associated with smoking cessation counseling among a national sample of 579 women pediatricians. Fifty-two percent of women pediatricians had received at least some training in cessation counseling and 41% counseled smoking patients at least once per year. Prevalence of counseling increased by amount of training; 20.7% of those with no training counseled at least once per year versus 62.0% of those with extensive training. Pediatricians 50-70 years of age were 1.8 times as likely as those 30-39 years of age to perform frequent counseling (p<0.01). Programs to promote smoking cessation training and counseling among pediatricians are needed. PMID- 12086201 TI - Continuous pH monitoring using the Paratrend 7 inserted into a peripheral vein in a patient with shock and congenital lactic acidosis. AB - The authors present a 25-year-old woman who was admitted to the ICU for treatment of shock, respiratory failure, and acidosis related to congenital lactic acidosis from pyruvate dehydrogenase deficiency. To aid in ongoing management of the metabolic acidosis, the Paratrend blood gas monitoring sensor was inserted through a peripheral venous site to provide a continuous measurement of pH and partial pressure of carbon dioxide (Pco2). With the venous insertion of the Paratrend, a clinically useful correlation with arterial blood gas values was noted. Linear regression analysis of the pH values from the venous blood gas analyses and the Paratrend monitor revealed r2 = 0.71 with p = 0.001 and r2 = 0.78 with a p = 0.0003 for the Pco2 values. Our preliminary experience suggests that venous placement of the Paratrend monitor can be used to provide clinically useful, continuous measurement of pH and Pco2. PMID- 12086202 TI - Gross hematuria in a young child with axillary lymphadenopathy. PMID- 12086203 TI - Erythropoietin deficiency causes anemia in nephrotic children with normal kidney function. PMID- 12086204 TI - Minimal change nephrotic syndrome in children with intrauterine growth retardation. PMID- 12086205 TI - Efficacy and safety of alendronate for the treatment of osteoporosis in diffuse connective tissue diseases in children. PMID- 12086206 TI - Nonaccidental methadone poisoning. PMID- 12086207 TI - New Doppler echocardiographic applications for the evaluation of early alterations in left ventricular diastolic function after coronary angioplasty. AB - AIMS: The early effect of percutaneous transluminal coronary angioplasty on left ventricular diastolic dysfunction was investigated with the aid of new Doppler echocardiographic applications. METHODS: Thirty patients with isolated severe left anterior descending coronary artery stenosis were included. All patients exhibited abnormal diastolic function demonstrated by prolonged isovolumic relaxation and deceleration time and decreased E/A ratio. New Doppler indexes included the flow propagation velocity of E wave, E and A waves' transit time to the left ventricular outflow tract, the ratio of these transit times in addition to the traditional isovolumic relaxation time, mitral deceleration time, and early and late transmitral peak flow velocities. All measurements were performed within 4 h before angioplasty and repeated within 24 h after the procedure. RESULTS: After angioplasty none of these parameters, except the A-wave transit time, were changed significantly. The A-wave transit time increased significantly from 57 +/- 5 ms to 78 +/- 7 ms within 24 h after successful angioplasty. The ratio of E- and A-wave transit time decreased significantly due to this significant increase in A-wave transit time. CONCLUSION: In this study, early improvement of Doppler index of left ventricular diastolic compliance is demonstrated after successful angioplasty. PMID- 12086208 TI - Dobutamine stress echocardiography and thallium-201 SPECT for detecting ischaemic dilated cardiomyopathy in patients with heart failure. AB - AIMS: A diagnosis of ischaemic aetiology of a dilated cardiomyopathy has important therapeutic and prognostic implications. In such patients, abnormal ECG and atypical symptoms limit the usefulness of standard ECG-ergometry in detecting myocardial ischaemia. To assess the values of high-dose dobutamine stress echocardiography and of Thallium-201 SPECT (exercise-reinjection-rest protocol) in differentiating between ischaemic and non-ischaemic dilated cardiomyopathy, 37 patients with suspected myocardial ischemia, low ventricular ejection fraction (23 +/- 5%) and heart failure were studied. METHODS AND RESULTS: Coronary artery disease was defined as >50% coronary stenosis in at least one coronary artery. By dobutamine stress echocardiography, ischaemic dilated cardiomyopathy was considered present when either an ischaemic response (biphasic response or direct deterioration) or a scar (fixed dyssynergy) was documented in at least two segments. By Thallium-201 SPECT, severe perfusion defects, either reversible (ischaemia) or fixed (scar), in at least two segments were considered markers of ischaemic dilated cardiomyopathy. Twenty-three patients had ischaemic dilated cardiomyopathy, while 14 had normal coronary arteries. The presence of myocardial ischaemia and/or scar by dobutamine stress echocardiography identified patients with ischaemic dilated cardiomyopathy with a sensitivity of 100% and a specificity of 86%. The sensitivity of Thallium-201 SPECT was 92%, its specificity was 69%. Three of the four false positive results occurred in patients with left bundle branch block. Thirty-two patients were concordantly classified by the two techniques (agreement=86%, k=0.73). CONCLUSION: Both dobutamine stress echocardiography and Thallium-201 SPECT are sensitive techniques for detecting the ischaemic aetiology of dilated cardiomyopathy. The specificity is lower, particularly by SPECT, when left ventricular branch block is present. PMID- 12086209 TI - The effect of aortic valve replacement on left ventricular mass assessed by echocardiography. AB - AIMS: The aim of the study was to investigate whether aortic valve replacement (AVR) has different effects on the left ventricular mass (LVM) in patients with different types of aortic valve disease, i.e. aortic stenosis (AS), aortic regurgitation (AR), or combined disease. METHODS AND RESULTS: We studied 100 patients with AS; (n=57), AR; (n=22), and combined disease; (n=21). Each patient was evaluated before and up to 8.7 years (mean follow-up period 1.42 +/- 1.65 years) after operation by transthoracic echocardiography. LVM was calculated based on echocardiographic parameters. Following AVR, the LVM for the entire group decreased from 373 +/- 145 g to 280 +/- 102 g (P=0.0001). The AS subgroup lost 13.7 +/- 30.7% from the preoperative LVM, compared to 28.8 +/- 24.8% in the AR subgroup (P<0.05) and 23.1 +/- 33.1% in the combined subgroup. The preoperative LVM was higher among patients with AR vs. AS patients (501 +/- 173 g vs. 319 +/- 100 g respectively, P<0.05). The decrease in LVM was significantly higher in patients with preoperative AR compared to AS patients (165 +/- 150 g vs. 58 +/- 96 g, respectively, P<0.05). However, the postoperative LVM remained higher among the former. The only preoperative parameter (of the LVM formula) that was different between AR and AS patients was the left ventricular end diastolic diameter (68 +/- 11 mm vs. 51 +/- 7 mm, respectively, P<0.05). No significant differences were found in wall thickness among the groups studied. Univariate analysis showed that gender (male), type of aortic valve disease (AR vs. AS), and larger prosthetic valve diameter correlated significantly with greater decrease in postoperative LVM. In multivariate analysis comparing the above variables between AS and AR subgroups, only the patient's gender (i.e. male) was identified as an independent predictor of LVM regression. CONCLUSION: Patients with AR have a greater decline in LVM compared with patients with AS following AVR. The postoperative decline in LVM is higher among males and those with larger prosthetic valve diameter. PMID- 12086210 TI - Impact of second harmonic imaging on the determination of the global and regional left ventricular function by 2D echocardiography: a comparison with MIBI gated SPECT. AB - AIMS: To evaluate the impact of second harmonic (SH) compared to fundamental mode (FM) imaging on the echocardiographic determination of ejection fraction (EF) and wall motion score index (WMSI), using MIBI gated SPECT as an independent reference. METHODS: Sixty-two consecutive patients underwent an echocardiography study and a MIBI gated SPECT over 24 hours. EF was estimated visually (estimated E) and was calculated with the Simpson biplane method (Tracing-T), for both FM and SH. WMSI was determined by two independent echo-readers blinded to the nuclear imaging results. The same segmentation and scoring system was used for WMSI determined by MIBI gated SPECT. RESULTS: The percentages of unscored segments because of suboptimal endocardial border detection were 19.5% (FM) and 9.0% (SH). The correlation coefficients (r) between SPECT-EF and echo-EF were: FM (E)=0.705, FM (T)=0.546, SH (E)=0.771, SH (T)=0.743. Agreement between SPECT-EF and echo-EF was acceptable for both imaging modalities (mean of the difference +/ 2 S.D.): -2.8 +/- 18.5 (FM) and -3.5 +/- 16.4 (SH). Correlation coefficients (r) between WMSI calculated by SPECT and by echo were 0.715 (FM) and 0.789 (SH). Agreement between SPECT-WMSI and echo-WMSI was good for all imaging modes but better with SH compared to FM: 0.12 +/- 0.91 (FM), 0.10 +/- 0.77 (SH). The interobserver correlation coefficients (r) for the WMSI were 0.939 (FM) and 0.996 (SH). The agreement between the two observers was better for SH compared to FM. The systematic differences (mean differences) were 0.21 (FM) and -0.01 (SH), and the random differences between both observers (2 S.D.) decreased from 1.55 (FM) to 0.29 (SH). CONCLUSIONS: The use of SH echocardiography decreases the number of unscored segments. This results in an important gain in correlation and agreement for EF determination between echo and SPECT, and in a considerable decline of the interobserver variability for the echo-determined WMSI. WMSI determined by MIBI gated SPECT correlated closely with the SH WMSI, and agreement between both methods was excellent. PMID- 12086211 TI - Relationship between left atrial appendage function and plasma concentration of atrial natriuretic peptide. AB - BACKGROUND: It has been reported that the most intensely granuled cardiocytes secreting atrial natriuretic peptide (ANP) are located in the atrial appendages. AIMS: To evaluate the mechanisms of ANP release in congestive heart failure. METHODS AND RESULTS: The relationship between ANP and left atrial appendage (LAA) function was evaluated in 36 patients who underwent both transoesophageal echocardiography and cardiac catheterization. ANP level correlated positively with mean pulmonary capillary wedge pressure (mPCWP; r=0.75, P<0.0001), whereas it showed no significant correlation with the mean right atrial pressure. mPCWP correlated positively with the maximal LAA area (LAAa; r=0.79, P<0.0001) and negatively with the LAA ejection fraction during atrial contraction (LAA-EF; r= 0.61, P<0.0001) and peak late diastolic LAA emptying flow velocity (LAAF; r= 0.69, P<0.0001). ANP level correlated negatively with the LAA-EF (r=-0.56, P<0.001) and with LAAF (r=-0.61, P<0.0001). ANP level correlated more closely with the LAAa (r=0.79, P<0.0001) than with maximal LA volume (r=0.34, P<0.05). Multiple stepwise regression analysis selected LAAa as the only factor independently related to the plasma concentration of ANP (ANP=-22.4+28.6 LAAa, r=0.79, P<0.0001). CONCLUSIONS: We conclude that the factor most predictive for ANP in patients with left-sided cardiac dysfunction is distension of the LAA wall rather than elevation in the LA pressure or distension of the body of LA. This is consistent with the known distribution of ANP-secreting cardiocytes. PMID- 12086212 TI - Doppler mirror image artifacts mimicking mitral regurgitation in patients with mechanical bileaflet mitral valve prostheses. AB - AIMS: To characterize the origin and mechanisms of generation of Doppler mirror images in the left atrium observed in patients with mechanical bileaflet mitral valve prosthesis. We hypothesized that these systolic colour Doppler images are artifactual and generated by reflection of the left ventricular outflow tract flow at the leaflet of the mitral valve prosthesis. METHODS AND RESULTS: Fifty patients with mechanical bileaflet mitral valves were prospectively examined by means of transthoracic Doppler-echocardiography. From different apical views, the left atrium was carefully interrogated for systolic colour flow signals, which were further analysed by pulsed wave Doppler. In all patients, a systolic colour Doppler signal was detected in the left atrium. Pulsed wave Doppler analysis revealed a low velocity signal (= mirror image) corresponding to the shape and duration of the left ventricular outflow tract signal. The peak velocities of these mirror images, however, were consistently lower than the velocities obtained from the left ventricular outflow tract. In addition, if paravalvular mitral valve leakage was present, a high velocity signal identical to the duration of mitral regurgitation time was detected outside the ring of the prosthesis. CONCLUSIONS: Mirror image artifacts mimicking mitral regurgitation occur in patients with mechanical mitral valves prosthesis. As mirror image artifacts can be specifically identified by analysis of shape. velocity, and duration of pulsed wave Doppler spectra, their identification will help to avoid misinterpretation of both spectral and colour Doppler signals in patients with prosthetic mitral valves. PMID- 12086213 TI - Influence of age and cardiovascular factors on regional pulsed wave Doppler myocardial imaging indices. AB - AIMS: To describe the influence of age and other cardiovascular factors on regional pulsed wave Doppler myocardial imaging (DMI), and to compare DMI with conventional transmitral echocardiography and the atrioventricular plane displacement (AVPD) method. METHODS AND RESULTS: Eighty-eight healthy subjects aged 20-81 years were examined by DMI, performed in the intraventricular septum just below the mitral annulus and in the corresponding lateral region, by transmitral pulsed wave Doppler echocardiography, and by AVPD. The DMI peak velocity during the left ventricular (LV) early filling phase (e), decreased with age from 12.3 +/- 2.3 cm/s in the youngest to 7.0 +/- 1.7 cm/s in the oldest tercentile (r=- 0.76, P<0.001). The DMI peak velocity during atrial contraction (a), increased from 7.5 +/- 2.2 cm/s in the youngest to 9.7 +/- 1.7 cm/s in the oldest tercentile (r=0.41, P<0.001). The DMI systolic peak velocity (s), decreased with age from 8.2 +/- 1.1 (youngest tercentile) to 6.9 +/- 1.1 (oldest tercentile), r=-0.39, P<0.001 cm/s, while the fraction shortening of the LV increased from 33.7 +/- 4.1 to 38.2 +/- 5.9% (r=0.36, P<0.01). The DMI e/a correlated with the transmitral early/atrial (E/A) (r=0.83, P<0.001) and with the AVPD measurement of diastolic function AV-LA/AV-mean (r=0.82, P<0.001). The DMI e velocity correlated with the transmitral E velocity (r=0.38, P<0.001). In the multiple regression analysis of DMI e, age was the strongest factor and LV mass index correlated inversely and independently with e. No DMI variables were influenced by gender, while transmitral E correlated with gender. The LV dimension variables explained 35% (R2 adjusted) of the DMI e velocity changes; only 7% of the transmitral E changes were explained by those variables. CONCLUSION: Regional DMI indices are highly age-dependent. In comparision with conventional echocardiography, regional DMI might be more influenced by LV geometry and by myocardial structural changes. These findings suggest a complementary role for regional DMI to conventional echocardiography for the assessment of myocardial function. PMID- 12086214 TI - New digital measurement methods for left ventricular volume using real-time three dimensional echocardiography: comparison with electromagnetic flow method and magnetic resonance imaging. AB - AIM: The aim of this study was to investigate the feasibility and accuracy of using symmetrically rotated apical long axis planes for the determination of left ventricular (LV) volumes with real-time three-dimensional echocardiography (3DE). METHODS AND RESULTS: Real-time 3DE was performed in six sheep during 24 haemodynamic conditions with electromagnetic flow measurements (EM), and in 29 patients with magnetic resonance imaging measurements (MRI). LV volumes were calculated by Simpson's rule with five 3DE methods (i.e. apical biplane, four plane, six-plane, nine-plane (in which the angle between each long axis plane was 90 degrees, 45 degrees, 30 degrees or 20 degrees, respectively) and standard short axis views (SAX)). Real-time 3DE correlated well with EM for LV stroke volumes in animals (r=0.68-0.95) and with MRI for absolute volumes in patients (r values=0.93-0.98). However, agreement between MRI and apical nine-plane, six plane, and SAX methods in patients was better than those with apical four-plane and bi-plane methods (mean difference = -15, -18, -13, vs. -31 and -48 ml for end diastolic volume, respectively, P<0.05). CONCLUSION: Apically rotated measurement methods of real-time 3DE correlated well with reference standards for calculating LV volumes. Balancing accuracy and required time for these LV volume measurements, the apical six-plane method is recommended for clinical use. PMID- 12086215 TI - High frame rate myocardial integrated backscatter. Does this change our understanding of this acoustic parameter? AB - AIMS: Integrated backscatter (IB) and its cyclic variation (CV) derived from radio-frequency (RF) data have been used as parameters to attempt myocardial tissue characterization. Prior imaging systems used to measure IB and its CV typically acquired data at frame rates of 20-30 Hz and at a resolution of 6-8 bits. If changes in IB levels are in part related to specific short-lived events, occurring within the cardiac cycle, this frame rate and resolution could have been too low to resolve adequately what might be a more complex data set. METHODS AND RESULTS: To investigate this possibility, we acquired real time two dimensional (2D) myocardial IQ data (the 'in-phase quadrature' sampled RF data) at high frame rate (> 100 Hz), high dynamic resolution (theoretical 19-bit) and a sector angle of 20 degrees. Several consecutive heart cycles of myocardial data were acquired from individual cardiac walls in five closed chest dogs and 10 healthy, young volunteers at normal heart rates. On the reconstructed RF data regions of interest were indicated, and IB and its CV were calculated. The extracted high frame rate curves showed that the CV of IB is not a smooth sinusoidal-like curve, but is made up of multiple reproducible peaks and troughs with local minima and maxima which are temporally related to active or passive mechanical events, i.e. systolic contraction, early ventricular relaxation and ventricular filling due to atrial contraction. CONCLUSIONS: This study shows that increasing the rate of real-time RF data acquisition results in a more complex, reproducible IB curve. The resolved maxima and minima in IB levels are related to specific phases of the myocardial contraction. Furthermore, spectral analysis showed that IB curves acquired at normal heart rates contain information up to 40 Hz. Hence, cardiac imaging data sets used to analyse regional myocardial function obtained at frequencies lower than 80 frames per second can contain aliased information. PMID- 12086216 TI - Utility of the proximal jet width in the assessment of regurgitant and stenotic orifices--effect of low velocity filter and comparison to actual vena contracta width: an in vitro and numerical study. AB - AIM: The colour Doppler proximal jet width (CDPJW) has been shown to be directly related to the severity of regurgitant and stenotic valve lesions. It is generally assumed that the CDPJW is equivalent to the vena contracta width (VCW). The purpose of this numerical and in vitro study was to evaluate how changing low velocity filter (LVF) settings on colour Doppler imaging devices may affect the CDPJW and its estimate of the VCW. METHODS: Computational fluid dynamic software was used to create models of round orifices (0.785, 1.13, 1.76, 3.14 cm2) at set flow rates (0.37-25 1/min). In vitro experiments were performed with round orifices (0.2, 0.95 and 1.76 cm2) with set flow rates (1.8-3.6 1/min). Laser flow visualization was used to obtain gold standard vena contracta widths for comparison to CDPJW for various LVF settings (4-24 cm/s). RESULTS: With the LVF set 'too low', overestimation errors occur. In contrast, with the LVF set 'too high', underestimation errors occur. Optimal LVF settings are required to avoid over- and underestimation errors of up to 280%. SUMMARY: The VCW is related to regurgitant or stenotic lesion severity, and the CDPJW is an approximation of the VCW. The CDPJW closely resembles the actual VCW only at optimally chosen LVF settings. LVF settings can have a significant impact on the accuracy of the CDPJW. Inter mediate filter settings remove unnecessary background noise while maintaining actual flow regions, thereby providing the best agreement between the CDPJW and the VCW. If treatment decisions are to be based on these measurements, understanding such dependencies becomes quite important. PMID- 12086217 TI - Left and right atrial volume by freehand three-dimensional echocardiography: in vivo validation using magnetic resonance imaging. AB - INTRODUCTION: Measurement of left and right atrial size is important for the management of arrhythmias, valvular and congenital heart disease. We have demonstrated that freehand three-dimensional (3D) echocardiography is more accurate and reproducible than two-dimensional (2D) echocardiography for measurement of left ventricular mass and volume. However, no prior study has validated the accuracy of freehand 3D for the determination of left or right atrial volume. METHODS: End-systolic (maximum) left and right atrial volumes were determined in 21 volunteer patients and normal subjects by one, two, and freehand 3D transthoracic echocardiography and compared to volumes obtained by gradient recalled magnetic resonance imaging. Three-dimensional echocardiographic determination of atrial volume was obtained using an acoustic spatial locator, a line-of-intersection display, and a surface reconstruction algorithm. Two dimensional echocardiographic atrial volumes were obtained from apical biplane images of the left atrium and an apical single plane image of the right atrium using a summation of disks method. One-dimensional (ID) estimates of left atrial volume were determined by cubing the M-mode ID antero-posterior dimension obtained on the parasternal long axis view. RESULTS: An excellent correlation was Obtained between freedhand 3D echocardiography and magnetic resonce imaging (MRI) for the left atrium (r = 0.90, SEE=9.6 ml) and for the right atrium (r = 0.91, SEE = 8.8 ml) with a small bias (left atrium 5.25 ml, right atrium 12.06 ml) and narrow limits of agreement (left atrium 22.14 ml, right atrium 25.54 ml). Two dimensional echocardiography correlated less well (left atrium r = 0.87, SEE = 10.23 ml, right atrium r = 0.79, SEE = 19.74 ml), and had a higher bias (left atrium 14.46 ml, right atrium 8.99 ml) and larger limits of agreement (left atrium 24.37 ml, right atrium 41.16 ml). One-dimensional estimates of left atrial volume correlated poorly with magnetic resonance determined left atrial volume (r = 0.80, SEE = 6.61 ml) and had unacceptably high bias (45.09 ml) and limits of agreement (35.52 ml). Interobserver variability was lowest for 3D echocardiography (left atrium 7.2 ml, 11%, right atrium 8.7 ml, 16%). CONCLUSIONS: Freehand 3D echocardiography using the line of intersection display for guidance of image positioning and a polyhedral surface reconstruction algorithm is a valid, accurate, reproducible method for determining left and right atrial volume in humans that is comparable to magnetic resonance imaging and is superior to current ID and 2D echocardiographic techniques. PMID- 12086218 TI - Subxiphoid pericardiocentesis guided by contrast two-dimensional echocardiography in cardiac tamponade: experience of 110 consecutive patients. AB - AIMS: We evaluated echo-guided pericardiocentesis with contrast study in cardiac tamponade management. PATIENTS AND METHODS: From 1982 to 1998 we performed pericardiocentesis in 110 patients (56 +/- 14 years old). Subxiphoid approach was used in 109. Cardiac tamponade was idiopathic (n = 16), secondary to malignant disease (n = 50) and miscellaneous disorders (n = 44). RESULTS: Pericardial fluid was bloody (n=75), serous (n = 29) or turbid (n = 6). Mean volume of fluid removed was 585 +/- 370 ml. When prolonged drainage (60 +/- 26 h) was used (n = 41), total effusion volume was 850 +/- 340 ml. Eleven deaths were observed during the early period following pericardiocentesis. No relation with procedure was demonstrated by autopsy in 10, and death always occurred in critically-ill patients (five malignant diseases, five cardiac ruptures and one septic shock). Other complications were: right ventricular puncture (n = 11) with deleterious effect in one, vasovagal hypotension (n = 6) and paroxysmal arrhythmia (n = 6). Surgical drainage was mandatory in 19 patients. It had to be done as an emergency (within 6 h), because of failure of the procedure in four patients. In 14 patients without prolonged drainage a delayed surgical evacuation was indicated, because of persistent (n = 3) or recurrent (n = 11) cardiac tamponade. Only one surgical procedure was required after prolonged drainage. CONCLUSIONS: Echo guided pericardiocentesis with contrast study is an effective technique which reduces the risk of cardiac tamponade management. It should be considered in patients with critical haemodynamic condition or advanced malignancy, and in patients with poor short-term prognosis. PMID- 12086219 TI - Rapid diagnosis and management of thoracic aortic dissection and intramural haematoma: a prospective study of advantages of multiplane vs. biplane transoesophageal echocardiography. AB - AIMS: The purposes of this study were to compare the accuracy of multiplane vs. biplane transoesophageal echocardiography (TEE) in the diagnosis of aortic dissection and aortic intramural haematoma, and to test whether these techniques provide all the diagnostic information required to make management decisions. METHODS AND RESULTS: Fifty-eight consecutive patients with clinically suspected aortic dissection were studied with multiplane TEE; all cases who required surgery underwent intraoperative monitoring with multiplane TEE. The following multiplane TEE data were analysed: the angle between current and 0 degrees plane at which each view was obtained; the success rate in the evaluation of true and false lumen, entry tear, coronary artery involvement, aortic regurgitation, pericardial effusion. Advantages of multiplane over biplane TEE have been evaluated by the demonstration of usefulness of views obtained in planes other than 0 degrees-20 degrees or 70 degrees-110 degrees, assuming that with manipulation of a biplane probe a 20 degrees arc could be added to the conventional horizontal and vertical planes. On the basis of TEE findings, aortic dissection was confirmed in 36 cases (18 type A, 12 type B, six intramural haematoma). The specificity and sensitivity of TEE in terms of the presence or absence of aortic dissection or intramural haematoma were 100%. An additional clinical value of multiplane over biplane TEE in the evaluation of ascending aorta, aortic arch, entry tears and coronary artery involvement was demonstrated. All cases with type A aortic dissection or intramural haematoma involving the ascending aorta had an operation that was performed immediately after the diagnosis (hospital mortality, 13%). Patients with type B aortic dissection were treated medically; 25% of these cases were operated later (hospital mortality, 0%). CONCLUSIONS: Multiplane and biplane TEE have excellent and similar accuracies in the evaluation of aortic dissection and intramural haematoma. Multiplane TEE improves the visualization of coronary arteries, aortic arch and entry tears; it appears to be an ideal method as the sole diagnostic approach before surgery in type A aortic dissection. PMID- 12086220 TI - Panic attacks in psychosis. AB - OBJECTIVE: To determine the prevalence, psychiatric comorbidity and suicidal ideation associated with co-occurring non-affective psychosis and panic attacks, compared with non-affective psychosis without panic attacks, in the general population. METHOD: Data were drawn from the National Comorbidity Survey (n = 8098). Statistical analyses were used to identify differences in sociodemographic characteristics, psychiatric comorbidity, suicidal ideation, and familial psychopathology between those with non-affective psychoses with and without panic, as well as to identify correlates of non-affective psychosis and panic. RESULTS: Co-occurring panic attacks and non-affective psychosis, compared with non-affective psychosis without panic, was associated with elevated psychiatric comorbidity and increased odds of suicidal ideation. CONCLUSION: Consistent with clinical findings, these data suggest that panic attacks commonly co-occur with psychosis in the community. Results indicate that the co-occurrence of panic and psychosis is associated with increased likelihood of psychiatric comorbidity and suicidality. These findings need replication. PMID- 12086221 TI - The impact of depression on the well-being, disability and use of services in older adults: a longitudinal perspective. AB - OBJECTIVE: To study the impact of depression on the wellbeing, disability and use of health services of older adults. METHOD: Prospective community-based study, using a large (n = 2,200) sample of the elderly (55-85) in the Netherlands. Using a 3-year follow-up, the effect of depressive symptoms (CES-D) on disability, wellbeing and service utilization was assessed, controlling for competing need for-care (chronic physical illness, functional limitation and cognitive decline), enabling (partner status, size of the social network, social support and locus of control), and predisposing factors (age, sex and level of education). RESULTS: Depressive symptoms have considerable impact on the wellbeing and disability of older people and clear economic consequences caused by inappropriate service utilization. Compared with other need-for-care variables the impact of depression is weaker (service utilization), similar (disability) or stronger (wellbeing). CONCLUSION: The steeply rising prevalence of competing health risks in later life does not influence the significance of depression. PMID- 12086223 TI - Genetic epidemiology: bringing genetics to the population--the NAPE Lecture 2001. AB - OBJECTIVE: To present an overview of the status of genetics of mental disorders and to describe the role of genetic epidemiology in the future of the implementation of the human genome initiative. METHOD: Reviews evidence on familial recurrence risk for major mental disorders and approaches to identify genes for complex disorders. RESULTS: The next decade will witness shifts in approaches of both epidemiology and genetics to address sources of complexity of the mental disorders. Descriptive genetic epidemiology will evolve into analytic genetic epidemiology by shifting the key questions from estimation of the magnitude of mental disorders to identification of risk and protective environmental factors that may be informative for both etiology and prevention. Genetics research will expand to population-based studies for complex disorders and will employ designs and methods that incorporate sources of complexity. CONCLUSION: In summary, the next era of human genetics will witness major shifts in approaches to identify the genes underlying mental disorders. The contributions of genetic epidemiology to translate advances in molecular genetics to public health are discussed. PMID- 12086222 TI - Confirmatory factor analysis of DSM-IV borderline, schizotypal, avoidant and obsessive-compulsive personality disorders: findings from the Collaborative Longitudinal Personality Disorders Study. AB - OBJECTIVE: To test the diagnostic constructs implied by DSM-IV Axis-II personality disorders by examining relationships between different combinations of DSM-IV criteria. METHOD: Confirmatory factor analysis was used to test the borderline, schizotypal, avoidant and obsessive-compulsive personality disorder constructs in a large treatment-seeking sample (N= 668) from a multisite study. A model based on the three DSM-IV Axis II clusters was also tested. Both models were tested against a unitary 'generic' model constructed from four criteria sets combined. RESULTS: Goodness-of-fit for both the three-cluster and four disorder models was significantly better than the unidimensional model, and the four disorder model was significantly better than the three-cluster model. Results were replicated using data from 2-year follow-up obtained by interviewers blind to original Axis II diagnoses at baseline. CONCLUSION: Support is provided for the DSM-IV disorder-level classification for schizotypal, borderline, avoidant and obsessive-compulsive personality disorders in a treatment-seeking sample. PMID- 12086224 TI - Personality profile among primary care patients: experimenting with the Arabic IPDE ICD-10. AB - OBJECTIVE: To estimate the prevalence of personality disorder (PD) among primary health care (PHC) patients, and to investigate the characteristic features of the International Personality Disorder Examination, the ICD-10 module (IPDE ICD-10). METHOD: A sample (n = 158) PHC patients in Al Ain, United Arab Emirates (UAE) were interviewed by general practitioners (GPs) using the Arabic version of the IPDE ICD-10. RESULTS: Patients interviewed were 82 males and 76 females. Definite PD was identified in 12.7%. Prevalence rates among males and females were 9.8 and 15.8% consecutively. Commonest personality disorders were the schizoid (5.1%), anankastic (4.4%) and the emotionally unstable, borderline type (3.8%). Comorbidity of PD was manifested by 30% of definite PD patients. The mean dimensional scores of definite and probable PD patients were 5.6 and 4.3 consecutively. CONCLUSION: The IPDE-ICD-10 is useful but relatively time consuming with repetition and need of rephrasing in some items. Dimensional measurement proved essential. PMID- 12086225 TI - Comparing patterns and costs of schizophrenia care in five European countries: the EPSILON study. European Psychiatric Services: Inputs Linked to Outcome Domains and Needs. AB - OBJECTIVE: To compare service utilization and cost profiles of people with schizophrenia living in Europe in order to understand differences in treatment and care costs. METHOD: Cross-sectional samples were taken of people with schizophrenia in five European locations. Sociodemographic, clinical and service use data were collected via interviewer-administered questionnaires. Site specific unit costs were obtained, transformed subsequently into a single currency (UK pound). Multiple regression analyses were conducted. RESULTS: There were widespread and considerable differences between sites in service utilization patterns and associated costs. Higher needs, greater symptom severity and longer psychiatric history are associated with higher costs: quality of life and service satisfaction are not. Few differences were found between sites in patterns of association with cost. CONCLUSION: Comparative analyses of the use and cost of mental health services can highlight existing variations helpfully in service provision and uptake. Methodological consistency is required if meaningful conclusions are to be drawn from such comparative data. PMID- 12086226 TI - Age-related adverse drug reactions to clomipramine. AB - OBJECTIVE: The aim of this study was to determine whether age itself is a significant factor in predicting adverse drug reactions in depressed inpatients treated with clomipramine. METHOD: The study involved 150 hospitalized, depressed patients treated with 150 mg clomipramine per day. Changes in orthostatic blood pressure during treatment as well as the patients' complaint about side-effects was examined with regard to age. The sample was divided into younger (<56) and older (56-70) groups. RESULTS: No significant differences between younger and older subjects were found on any of the 44 side-effects recorded. However, older depressed patients suffer from more pronounced orthostatic hypotension than younger patients. CONCLUSION: Older depressed patients who have been treated with clomipramine suffer from more severe orthostatic hypotension than younger patients. However, with the right precautions it is safe to treat older patient up to the age of 70 years with a tricyclic antidepressant. PMID- 12086227 TI - Leukocytosis and hypoalbuminemia in mixed bipolar states: evidence for immune activation. AB - OBJECTIVE: Although activation of an immune response during major depressive episodes has been reported, less is known about changes during manic and mixed bipolar episodes. METHOD: Albumin and leukocyte levels were compared between subjects in manic and mixed bipolar episodes. Neutrophil, lymphocyte and monocyte levels were compared between the two groups. RESULTS: Albumin levels were lower in mixed manic subjects as opposed to pure manic subjects and in the combined groups levels were lower in females than in males. Leukocyte levels were higher in mixed manic patients compare with pure manic patients. Both neutrophil and monocyte levels were higher in the mixed manic patients but lymphocyte levels were no different. CONCLUSION: Leukocytosis and hypoalbuminemia during mixed manic states suggest immune activation in mixed mania similar to depression. This finding also tends to support the recognition of mixed mania as a distinct bipolar state. PMID- 12086228 TI - Involvement of plasma atrial natriuretic peptide in protracted alcohol withdrawal. AB - OBJECTIVE: Atrial natriuretic peptide (ANP) has been shown to inhibit the effects of corticotrophin releasing hormone, corticotrophin and cortisol, and to influence affective and anxiety symptoms in man. We tested the hypothesis of whether ANP is associated with endocrine and psychopathological disturbances during acute alcohol withdrawal. METHOD: ANP and cortisol plasma concentrations were studied in alcoholics during in-patient detoxification and in healthy controls. Additionally, craving, depressive mood and anxiety were assessed. RESULTS: Although mean ANP levels increased significantly in alcoholics between days 1 and 14, they remained diminished compared to controls. Separating a subgroup of alcoholics with a decrease of ANP levels during withdrawal, these individuals revealed significantly elevated scores for mean and maximum craving and a trend to an elevated self-rated anxiety on day 14. CONCLUSION: We suggest that a dysregulation of ANP plasma levels during alcohol withdrawal may contribute to symptoms of protracted withdrawal such as craving and anxiety. PMID- 12086229 TI - Apolipoprotein E genotype and schizophrenia: further negative evidence. AB - OBJECTIVE: To investigate the association between apolipoprotein E (ApoE) genotype and schizophrenia. METHOD: We genotyped 106 schizophrenic out-patients [Diagnostic Statistic Manual IV (DSM-IV) criteria] and 250 healthy volunteers (hospital staff and blood donors) from Asturias (Northern Spain). The ApoE genotypes (epsilon2, epsilon3, epsilon4-alleles) were determined after polymerase chain reaction (PCR) amplification, followed by digestion with the restriction enzyme Cfol and electrophoresis on a 4% agarose gel. RESULTS: No significant differences in ApoE-allele frequencies between patients and controls was found, although an increased 64-frequency was recorded in patients compared with controls [9.0% vs. 6.2%, P = 0.124; odds ratio (OR) = 1.49; 95% confidence interval (CI) = 0.82-2.70]. ApoE-genotype frequencies did not differ between both groups. The mean age of onset for schizophrenic patients that carried the epsilon4-allele was not significantly different from that of patients without this allele. CONCLUSION: Variation in the ApoE gene was not associated with the development of schizophrenia in our population. ApoE-genotypes did not modify the age of onset of the disease. PMID- 12086230 TI - Herbal products and serious side effects: a case of ginseng-induced manic episode. AB - OBJECTIVE: Ginseng root extract is a widely used herbal product not devoid of side effects. This report describes the development of manic symptoms after ginseng consumption in a patient with affective disorder. Other potentially harmful side effects of ginseng are also reviewed. METHOD: A single case report. RESULTS: A 56-year-old woman with previous affective disorder presented a manic episode during ginseng intake. Symptoms disappeared rapidly with low doses of neuroleptics and benzodiazepines after ginseng suppression. CONCLUSION: Ginseng may produce manic symptoms. A special risk situation seems to be affective patients under antidepressant medication. The case emphasizes the lack of harmlessness of herbal products. Patients should be routinely asked about the use of herbal products and diet supplements. PMID- 12086231 TI - Coronary artery calcification and end-stage renal disease: vascular biology and clinical implications. PMID- 12086232 TI - Cardiovascular mortality in chronic renal failure: hyperphosphatemia, coronary calcification, and the role of phosphate binders. PMID- 12086233 TI - Noninvasive quantification of coronary artery calcification: methods and prognostic value. PMID- 12086234 TI - Simple dimple rule for sacral dimples. PMID- 12086235 TI - Management guidelines for sport-related concussions. PMID- 12086236 TI - Initiating discussions about advance directives: the family physician's role. PMID- 12086237 TI - Management of common arrhythmias: Part I. Supraventricular arrhythmias. AB - Family physicians frequently encounter patients with symptoms that could be related to cardiac arrhythmias, most commonly atrial fibrillation or supraventricular tachycardias. The initial management of atrial fibrillation includes ventricular rate control to provide adequate cardiac output. In patients with severely depressed cardiac output and recent-onset atrial fibrillation, immediate electrical cardioversion is the treatment of choice. Hemodynamically stable patients with atrial fibrillation for more than two days or for an unknown period should be assessed for the presence of atrial thrombi. If thrombi are detected on transesophageal echocardiography, anticoagulation with warfarin for a minimum of 21 days is recommended before electrical cardioversion is attempted. Patients with other supraventricular arrhythmias may be treated with adenosine, a calcium channel blocker, or a short-acting beta blocker to disrupt reentrant pathways. When initial medications are ineffective, radiofrequency ablation of ectopic sites is an increasingly popular treatment option. PMID- 12086238 TI - Management of common arrhythmias: Part II. Ventricular arrhythmias and arrhythmias in special populations. AB - In patients without established cardiac disease, the occurrence of premature ventricular complexes without sustained ventricular tachycardia is more an annoyance than a medical risk, and treatment is not required. In contrast, patients with established heart disease and premature ventricular complexes have a higher likelihood of developing ventricular tachycardia or fibrillation. These patients should be treated with a beta blocker or class I antiarrhythmic drug. Treatment of arrhythmias in pregnant women is rarely needed. When treatment is required, amiodarone should be avoided, and beta blockers should be used with caution, because these agents have been associated with fetal growth retardation. The most important rhythm abnormality in athletes is ventricular tachycardia associated with hypertrophic cardiomyopathy. If the presence of the disease is confirmed by echocardiography, beta-blocker therapy is necessary, and these patients should be limited to participation in nonstrenuous sports. Acute arrhythmias in children with Wolff-Parkinson-White syndrome can be treated with adenosine. Radiofrequency ablation of the accessory pathway can provide long-term control. PMID- 12086239 TI - Basal cell nevus syndrome: guidelines for early detection. AB - Basal cell nevus syndrome is an autosomal dominant condition with complete penetrance and variable expressivity. It is characterized by five major components, including multiple nevoid basal cell carcinomas, jaw cysts, congenital skeletal abnormalities, ectopic calcifications, and plantar or palmar pits. Other features include a host of benign tumors, ocular defects, and cleft lip and palate. Guidelines for diagnosis include a family history, careful oral and skin examinations, chest and skull radiographs, panoramic radiographs of the jaw, magnetic resonance imaging of the brain, and pelvic ultrasonography in women. PMID- 12086241 TI - Information from your family doctor. How to prepare for pregnancy. PMID- 12086240 TI - Preconception health care. AB - Appropriate preconception health care improves pregnancy outcomes. When started at least one month before conception, folic acid supplements can prevent neural tube defects. Targeted genetic screening and counseling should be offered on the basis of age, ethnic background, or family history. Before conception, women should be screened for human immunodeficiency virus and syphilis infection and begin treatment to prevent the transmission of disease to the fetus. Immunizations against hepatitis B, rubella, and varicella should be completed, if needed. Women should be counseled on ways to prevent infection with toxoplasmosis, cytomegalovirus, and parvovirus B19. Environmental toxins such as cigarette smoke, alcohol, and street drugs, and chemicals such as solvents and pesticides should be avoided. In women with diabetes, it is important to optimize disease control through intensive management before pregnancy. Medications for hypertension, epilepsy, thromboembolism, depression, and anxiety should be reviewed and changed, if necessary, before the patient becomes pregnant. Counseling about exercise, obesity, nutritional deficiencies, and the overuse of vitamins A and D is beneficial. Physicians may also choose to discuss occupational and financial issues related to pregnancy and to screen patients for domestic violence. PMID- 12086242 TI - Guidelines for managing Alzheimer's disease: Part II. Treatment. AB - Once the clinical diagnosis of Alzheimer's disease has been made, a treatment plan must be developed. This plan should include cholinesterase inhibitor therapy to temporarily improve cognition or slow the rate of cognitive decline, management of comorbid conditions, treatment of behavioral symptoms and mood disorders, provision of support and resources for patient and caregiver, and compliance with state-mandated reporting requirements for driving impairment and elder abuse. The primary caregiver can be a valuable ally in communication, management of care, and implementation of the care plan. Patient symptoms and care needs change as Alzheimer's disease progresses. In the early stage of the disease, the family physician should discuss realistic expectations for drug therapy, solicit patient and family preferences on future care choices, and assist with advance planning for future care challenges. In the middle stage, the patient may exhibit behavioral symptoms that upset the caregiver and are difficult to manage. When the patient is in the advanced stage of Alzheimer's disease, the caregiver may need support to provide for activities of daily living, help in making a difficult placement decision, and guidance in considering terminal care options. Throughout the course of the disease, routine use of community resources allows care to be provided by a network of professionals, many of whom will be specialists in Alzheimer's disease. PMID- 12086243 TI - Screening for breast cancer: recommendations and rationale. PMID- 12086244 TI - Ingrown toenail removal. AB - Ingrown toenail is a common problem resulting from various etiologies including improperly trimmed nails, hyperhidrosis, and poorly fitting shoes. Patients commonly present with pain in the affected nail but with progression, drainage, infection, and difficulty walking occur. Excision of the lateral nail plate combined with lateral matricectomy is thought to provide the best chance for eradication. The lateral aspect of the nail plate is removed with preservation of the remaining healthy nail plate. Electrocautery ablation is then used to destroy the exposed nail-forming matrix, creating a new lateral nail fold. Complications of the procedure include regrowth of a nail spicule secondary to incomplete matricectomy and postoperative nail bed infection. When performed correctly, the procedure produces the greatest success in the treatment of ingrown nails. Basic soft tissue surgery and electrosurgery experience are prerequisites for learning the technique. PMID- 12086245 TI - Information from your family doctor. Ingrown toenail removal. PMID- 12086246 TI - AUA and ASRM produce recommendations for male infertility. American Urological Association, Inc and American Society for Reproductive Medicine. PMID- 12086247 TI - Measuring disease activity and outcomes in clinical studies. PMID- 12086248 TI - Diagnostic strategies in vasculitis affecting the central nervous system. PMID- 12086249 TI - Conventional treatment and outcome of Wegener's granulomatosis and microscopic polyangiitis. PMID- 12086250 TI - Treatment of giant-cell arteritis: where we have been and why we must move on. PMID- 12086251 TI - Henoch-Schonlein purpura (treatment and outcome). PMID- 12086252 TI - Targeted therapies in systemic vasculitis. PMID- 12086253 TI - Inflammation in acute coronary syndromes. PMID- 12086254 TI - Kidney transplantation and ANCA-associated vasculitis. PMID- 12086256 TI - New approaches to the management of subglottic stenosis in Wegener's granulomatosis. AB - Further development of conservative endoscopic procedures at the CCF has provided patients with dependable surgical adjuncts to the systemic medications of WG. In most instances, a single surgical dilatation procedure stabilizes the patients at an airway diameter that exceeds 50% of the norm, thereby rendering the patient almost asymptomatic at rest and minimally restricted during exercise. The more chronic patients presenting with history of previous surgical procedures performed on their larynx and trachea, usually in other institutions, have been observed to require consecutive treatments after the original treatment, enjoying shorter symptom-free intervals. We have not encountered any local complications such as damage to the vocal cords, altered voice, or compromised structural integrity of the larynx and the trachea. The procedure has been found to be effective and well-tolerated as a means for treating, maintaining, and rehabilitating patients with chronic airway obstruction, particularly in those that have been initially treated by our service, and those that were managed from the very beginning of their disease. However, even the more difficult and complicated cases clearly demonstrate an improvement in their condition through the above treatment protocol, and the interval between treatments gradually increases in this group as well. The more aggressive long-term tube-free tracheostomy procedure, usually performed only on difficult and select patients with severe complications, has proven itself to be a highly gratifying procedure, achieving a permanent mode of management for these patients which safely allows for almost complete freedom from symptoms combined with good tolerance and functional rehabilitation. Video documentation will serve to further demonstrate the beneficial effects of both these modes of treatment. PMID- 12086255 TI - Surgical treatment of Takayasu's disease. AB - Takayasu's disease affects young females in the second and third decade of life. During the chronic phase, the aorta and its major arteries become stenotic, causing significant sequelae. Surgical treatment is possible with expectation of good results. The author advises treatment of stenotic arteries that are potentially dangerous or that adversely affect lifestyle with either percutaneous transluminal angioplasty or surgery. The author emphasizes the difference between surgical procedures for atherosclerosis versus a procedure for Takayasu's disease. PMID- 12086257 TI - Known causes of vasculitis in man. PMID- 12086258 TI - Sinonasal complications of vasculitic diseases. PMID- 12086259 TI - HCV and cryoglobulinemic vasculitis. PMID- 12086260 TI - Infections in primary vasculitides. PMID- 12086261 TI - Pathogenic mechanisms in giant cell arteritis. AB - T lymphocytes, encountering stimulatory signals in the adventitia of medium-size arteries, emerge as the key players in inflammation-associated injury pathways. In GCA, all injury mechanisms have been related to effector macrophages. Regulated by IFN-gamma-producing T cells, macrophages commit to distinct avenues of differentiation and acquire a spectrum of potentially harmful capabilities (Figure 1). Macrophages in the adventitia focus on production of pro-inflammatory cytokines. Macrophages in the media specialize in oxidative damage with lipid peroxidation attacking smooth muscle cells and matrix components. These macrophages also supply reactive oxygen intermediates that, in combination with nitrogen intermediates, cause protein nitration of endothelial cells. Production of oxygen radicals is complemented by the production of metalloproteinases, likely essential in the breakdown of elastic membranes. With the fragmentation of the internal elastic lamina, the intimal layer becomes accessible to migratory myofibroblasts that, driven by PDGF, form a hyperplastic intimal layer and cause occlusion of the vessel lumen. Expansion of the hyperplastic intima is accompanied by intense neoangiogenesis, supported by angiogenesis factors that again derive from specialized macrophages. Similarities in injury pathways between GCA and another arterial disease, atherosclerosis, are beginning to be recognized. Specifically, activated T cells and macrophages are increasingly appreciated as key players in the process of instability and rupture of atherosclerotic plaque. A specialized subset of CD4 T cells, CD4+ CD28- T cells, are suspected to participate in tissue injury in the plaque. These T cells are equipped with cytolytic capabilities and release large amounts of IFN-gamma. Comparative studies between patients with GCA and those with acute coronary syndromes should enhance our ability to define the principles of arterial wall inflammation, the specifics of injury in that microenvironment, and help in the identification of the eliciting signals. PMID- 12086262 TI - Implications for pathogenesis of patterns of injury in small- and medium-sized vessel vasculitis. AB - The different pathologic features of different types of necrotizing vasculitis indicate that there are different pathogenic mechanisms causing the injury. The pathogenic mechanisms for medium-sized-vessel vasculitis are most effective at causing injury in arteries and are not effective at causing injury in smaller vessels. The predilection of medium-sized-vessel vasculitis for bifurcations may relate to the increased expression of adhesion molecules and increased numbers of intimal macrophages at these sites. The preferential involvement of small vessels by small-vessel vasculitis may relate to the requirement for close apposition between leukocytes and endothelial cells for the pathogenic mechanisms to be operational. The pathology of the necrotizing vasculitis of Kawasaki disease is most consistent with a primary role for monocytes/macrophages and T lymphocytes in the acute injury. The pathology of the necrotizing vasculitis of polyarteritis nodosa and small-vessel vasculitis, including ANCA-vasculitis, is most consistent with a primary role for neutrophils and monocytes in the acute injury. PMID- 12086263 TI - Granuloma formation, implications for the pathogenesis of vasculitis. AB - The pathogenesis of granulomatous inflammation is complex and involves a variety of mechanisms acting in concert to bring about an inflammatory lesion that is able to contain and destroy intracellular pathogens. While this process is crucial to host defense, it is also a two-edged sword in that excessive or inappropriate granulomatous inflammation results in considerable damage to normal tissue. In recent years, there has been significant progress in dissecting the immunologic events involved in granuloma formation and maintenance. A better understanding of these events will allow us to more precisely modulate the granulomatous inflammatory response to the benefit of patients with both infectious and autoimmune diseases. PMID- 12086264 TI - Endothelial cell biology, perivascular inflammation, and vasculitis. PMID- 12086265 TI - Understanding the pathogenesis of ANCA: where are we today? AB - The role of ANCA, ANCA antigens, endothelial cell damage, genetic and environmental pressures, and the "activatability" of leukocytes will probably prove to be important variables in human ANCA vasculitis. The advent of a reliable animal model may open new areas of investigation and treatment of these vasculitic conditions. PMID- 12086266 TI - ANCA subsets: influence on disease phenotype. PMID- 12086267 TI - Genetics of ANCA-associated vasculitides. PMID- 12086269 TI - Kawasaki disease: etiology, pathogenesis, and treatment. PMID- 12086268 TI - Classification of anti-endothelial cell antibodies into antibodies against microvascular and macrovascular endothelial cells: the pathogenic and diagnostic implications. PMID- 12086270 TI - Host and viral genes that control herpesvirus vasculitis. PMID- 12086271 TI - Epidemiology of giant-cell arteritis. PMID- 12086272 TI - Epidemiology of Wegener's granulomatosis, microscopic polyangiitis, and Churg Strauss syndrome. PMID- 12086273 TI - Epidemiology of Henoch-Schonlein purpura. PMID- 12086274 TI - Cytokines in giant-cell arteritis. AB - Cytokines are small proteins that serve as chemical messengers between cells, regulating cell growth and differentiation, tissue repair and remodeling, and many aspects of the immune response. Cytokines are instrumental in determining the nature, magnitude, and duration of inflammatory reactions and, as such, represent ideal targets for interfering with pathogenic processes. In OCA and PMR, cytokines are encountered in two locations, the inflammatory infiltrates accumulating in the arterial wall and in the circulation. IL-6, a cytokine involved in stimulating acute-phase responses, is located upstream of many of the laboratory abnormalities considered helpful in diagnosing and managing GCA/PMR, including elevated ESR and CRP. IL-6 has the potential to be helpful in predicting disease severity and may allow for a tailoring of immunosuppressive therapy. There is evidence suggesting that IL-6 outperforms other chemical markers in detecting disease activity and could, therefore, have a role in monitoring treatment. Interesting pathogenic clues have been derived from studies of cytokines produced in the vascular lesions. IFN-gamma has emerged as a key regulator in determining the nature and direction of the inflammatory response. IFN-gamma appears to be critically involved in modulating the process of intimal hyperplasia, the most destructive consequence of vasculitis, and, as such, emerges as a prime target for novel therapeutic approaches. PMID- 12086275 TI - Utility of imaging studies in assessment of vascular inflammation. PMID- 12086276 TI - Medical treatment and secondary prevention of coronary heart disease in general practice in Iceland. AB - OBJECTIVE: To evaluate the implementation of secondary prevention and treatment of coronary heart disease (CHD) in general practice in Iceland. SETTINGS: Two health care centers adjacent to Reykjavik with a total of 25766 inhabitants. PATIENTS: All patients (533) with CHD living in the study area were sent an invitation letter and a request for informed consent. Those who chose to participate answered a questionnaire about CHD risk factors and their current treatment, and their medical records were reviewed. The patients were divided into four groups on the basis of their history: I. Coronary artery bypass surgery (CABG), II. Percutaneous transluminal coronary angioplasty (PTCA), III. Myocardial infarction (MI), IV. Angina pectoris (AP). If a patient fulfilled the criteria for more than one diagnostic group the CABG group had the highest priority followed by PTCA, MI and finally AP. MAIN OUTCOME MEASURES: Blood pressure, smoking habits, BMI, exercise profile, cholesterol levels and drug therapy. RESULTS: Of 533 patients with CHD, 402 (75%) participated in the study, 15% were managed exclusively by their family physician and 23% by both cardiologists and family physicians. Obesity was relatively common, with nearly 60% being overweight (BMI > 25). Average cholesterol in the total group was 6.2 mmol/L (95% CI 6.07 to 6.34). Blood pressure had been recorded in 92% of the patients, and mean systolic and diastolic blood pressures were 143 and 82 mmHg, respectively. While 15% were current smokers, 56% were ex-smokers. A total of 113 patients (28%) were being treated with cholesterol-lowering drug therapy at the time of the study. Respective treatment ratios in the four subgroups were 47% in group I, 42% in II, 25% in III and 13% in group IV. Aspirin was taken by 284 patients (71%), beta blockers by 52% and calcium channel blockers by 36%. More than twice as many women than men were treated with nitrates, 57% versus 27%. CONCLUSIONS: The results indicate that there are numerous possibilities for improvements in secondary prevention and medical management of coronary heart disease in Iceland. Particular emphasis should be placed on smoking cessation, life-style modification with exercise and diet recommendations to lower BMI and lipid-lowering therapy. PMID- 12086277 TI - Do general practitioner and patient agree about the risk factors for ischaemic heart disease? AB - OBJECTIVE: To analyse agreement between patients' and GPs' perceptions of risk factors and overall risk of ischaemic heart disease (IHD). DESIGN: Cross sectional study based on paired information from patients and GPs. SETTING: Twenty-six GPs in the County of Ringkobing, Denmark, participating in a medical audit during 3 weeks in May 1999. SUBJECTS: 252 patients with IHD and 1239 without IHD. MAIN OUTCOME MEASURES: GPs and patients were asked about specific risk factors for IHD and their perception of overall risk. Their agreement was evaluated by Kappa statistics. RESULTS: Agreement between GPs and patients varied from 70% to 97%. Disagreement was observed most often for patients with IHD and patients listed with elderly GPs ( > 50 years). Generally, patients perceived the overall risk of IHD lower than their doctors, and for most patients with a perception of low risk the GP estimated the risk as high. CONCLUSIONS: Patients and GPs have different perceptions of the risk of [HD. This may be due to different perceptions of the importance of specific risk factors and different reference frames for risk perception. GPs have an important role in communicating the meaning of risk factors and interventions should be considered to improve risk communication in general practice. PMID- 12086278 TI - Better to prevent than to treat. PMID- 12086279 TI - Integration of information about cardiovascular risk factors: how do highly motivated women in a lifestyle intervention programme act and react? AB - OBJECTIVE: To understand the experience of being at risk and of participating in health interventions. How is information about risk factors handled on an individual level with respect to feeling, thinking and doing? DESIGN: Tape recorded, open interviews, written out and analysed using a descriptive/analytical method. SETTING: Selected women who had participated in a lifestyle intervention programme aimed at reducing risk factors for cardiovascular disease in Stromstad, a community of 10 000 inhabitants on the west coast of Sweden. SUBJECTS: Eight women in the most active group who had had some contact with the project during the whole period 1985-94. MAIN OUTCOME MEASURES: The creation of core concepts as a result of close text reading through codes and categories. RESULTS: Three core concepts in relation to the handling of risk factors were identified: there is no one but yourself to rely on, resisting invasion, and living with incompatibility, based respectively on the subjects' self-efficacy and self-awareness, their ways of maintaining a good life and their trying to understand the risk-factor concept. CONCLUSION: Risk-factor-oriented health interventions focus on disease and create uncertainty as to the relationship between the concepts of risk and disease. The powerful health resources demonstrated by the women in this study to counterbalance the risk pre occupation suggest changing to health-oriented interventions that focus on individual health resources. PMID- 12086280 TI - Time for a "vision zero" concerning premature death from ischaemic heart disease? AB - OBJECTIVE: To study the trend in premature mortality (before 75 years of age) from ischaemic heart disease (IHD) in a Swedish primary health care district compared to communities of similar demographic situation and all Sweden. DESIGN: Mortality from IHD in men and women was compared between the community of Habo and other Swedish communities of similar size and all Sweden for the period 1984 96. SETTING: The community of Habo in Sweden with about 9600 inhabitants. INTERVENTION: Intensified efforts by the local primary health care organisation, in co-operation with the community, in respect of primary and secondary cardiovascular prevention since the beginning of the 1980s. RESULTS: Mortality from IHD has decreased significantly both in Habo and in Sweden during these years. The decrease has been more prominent in Habo than in Sweden as a whole and other Swedish communities of similar demographic situation. CONCLUSION: With increased and purposeful efforts in primary and secondary prevention, in co operation between the community and primary health care, it is possible to substantially decrease mortality from IHD in the community. PMID- 12086281 TI - Incidence and management of intermittent claudication in primary care in The Netherlands. AB - OBJECTIVE: To study the incidence and management of intermittent claudication in primary care. DESIGN: In a longitudinal survey among 161 general practitioners (GPs) all incident diagnoses of intermittent claudication (IC) were selected and available information on GP management was studied. SETTING: Dutch National Survey of Morbidity and Interventions in General Practice. SUBJECTS: 74153 patients aged 55 years and over. MAIN OUTCOME MEASURES: Age- and sex-specific incidence rates. RESULTS: The overall incidence rate of IC was 6.4 per 1000 person years (95% confidence interval (CI) 5.3-7.7). Incidence rate in creased from 4.0 per 1000 person years in men aged 55-59 to 12.9 per 1000 person years in men aged over 85, and for women in the same age categories from 3.3 per 1000 person years to 8.2 per 1000 person years, respectively. Of the 117 incident cases of IC, 43 (37%) were referred to a hospital specialist. In 55 cases (47%), drugs were prescribed by the GP and in 101 cases (86%) the GP gave advice, notably pertaining to exercise and cessation of smoking. CONCLUSION: The incidence rate of IC in primary care is high in both men and women, and clearly increases with age. Only a minority of patients are referred to a hospital specialist, while the vast majority receive lifestyle advice. PMID- 12086282 TI - Parents' perspectives on otitis media and antibiotics. A qualitative study. AB - BACKGROUND: Judicious use of antibiotics is an important factor in decreasing the current increase in bacterial resistance. Discussion with patients and their guardians on whether to use antibiotics or not can be expected to increase in the future. OBJECTIVE: To explore the views and feelings of parents of children with acute otitis media towards the disease and its treatment. DESIGN: A qualitative study with semi-structured interviews. SUBJECTS: Twenty-one parents of pre-school children with newly diagnosed acute otitis media who had attended a primary care centre in Reykjavik. RESULTS: The parents did not consider the disease a threat. They generally thought the disease was caused by environmental factors or by a disturbance in body mechanism. The majority thought that treatment with antibiotics could harm their children. CONCLUSION: The parents of young children hold views on acute otitis media and antibiotics which are markedly different from the medical model. PMID- 12086283 TI - Diagnostic accuracy of acute maxillary sinusitis in adults in primary care. AB - OBJECTIVE: To evaluate the diagnostic value of symptoms and signs used in diagnosing acute community-acquired maxillary sinusitis (ACAMS), and to find useful clinical predictors for diagnosing it in adults in primary care. DESIGN: Prospective study in the study group and retrospective study in the reference group. SETTING: One primary care centre in the study group and another in the reference group. SUBJECTS: 50 adults with prolonged upper respiratory tract infection > or = 7 days) or self-suspected maxillary sinusitis. As a reference group, an analysis of records and diagnoses made by GP of 45 adult patients with the same inclusion criteria from another primary care clinic. MAIN OUTCOME MEASURES: The signs and symptoms associated with the diagnosis of ACAMS by GP and otorhinolaryngologist. The value of CRP, ultrasound and peak nasal expiratory flow in the diagnosis. RESULTS: Neither ultrasound nor any of the clinical signs increased the accuracy of diagnosis. Peak nasal expiratory flow below the normal range was associated with the diagnosis of ACAMS (p = 0.03). The presence of an otorhinolaryngologist had an influence on GP practice when compared with the reference group. CONCLUSION: The diagnosis of ACAMS is difficult with the available clinical methods. Peak nasal expiratory flow needs further study. New diagnostic means are needed. PMID- 12086284 TI - The relation between sale of antimicrobial drugs and antibiotic resistance in uropathogens in general practice. AB - BACKGROUND: Overuse of antimicrobial drugs has resulted in an alarming increase in bacterial resistance in most countries. The relevance for general practice is unknown. OBJECTIVE: To evaluate the impact of the sale of antimicrobial drugs on bacterial resistance as found in uropathogens from general practice. SETTING: General practice in Belgium and Norway. METHODS: Observational study. RESULTS: The sale of antimicrobial drugs indicated for use in the treatment of urinary tract infection was four times higher in Belgium than in Norway (18.5 vs 4.4 DDD/1000 inhabitants/day). The antibiotic resistance reported by microbiological laboratories as valid for general practice was significant higher in Belgium than in Norway (ampicillins (44% vs 27%), co-trimoxazole (28% vs 17%), fluoroquinolones (12% vs 2%) and nitrofurantoin (16% vs 11%, p < 0.0001 for all). However, the antibiotic resistance found in urine samples from dysuric women in general practice was similar (trimethoprim 14% vs 12%, co-trimoxazole 14% vs 11%, nitrofurantoin 7% vs 3%), except in the case of ampicillins (30% vs 19%, p < 0.05). CONCLUSION: The impact of the antimicrobial sale on resistance in uropathogens seems less than expected at the general practice level, even though local microbiological reports mention fairly high antibiotic resistance data. Adapted methods for following-up bacterial resistance evolution in general practice are needed. PMID- 12086285 TI - Influence of mild infections on iron status parameters in women of reproductive age. AB - OBJECTIVE: To investigate the effects of mild infections on iron status parameters. DESIGN: A prospective observational study. SETTING: A population of female nurse students in Norway. SUBJECTS: 33 women with self-reported episodes of infection and 33 women without infections but with comparable s-ferritin at baseline. MAIN OUTCOME MEASURES: Change from baseline in haemoglobin, s-iron, TIBC, transferrin saturation, s-ferritin, e-protoporphyrin and s-transferrin receptor on days 3, 7, 14, 21, 28 and 56 after onset of illness, compared to changes in non-infected subjects tested in parallel. RESULTS: In febrile illness, such as influenza, there was a significant rise in s-ferritin that could take more than a month to normalise. S-ferritin increased significantly when CRP rose above 20 mg/L, but a normal CRP could not preclude falsely high s-ferritin values due to infection. S-iron and transferrin saturation fell below normal range in a substantial proportion of cases in the symptomatic stage, even in infections without a febrile response, such as the common cold, but was normalised within a week after onset of infection. CONCLUSION: For more than a month after febrile illness such as influenza, s-ferritin is not a reliable measure for ruling out iron deficiency in women of reproductive age. PMID- 12086286 TI - Use of plasma methylmalonic acid in diagnosing vitamin B-12 deficiency in general practice. AB - OBJECTIVES: To examine the reasons why general practitioners (GPs) request plasma methylmalonic acid (MMA) tests and how they respond to a result above the reference interval. DESIGN: Retrospective study of medical records. SETTING: Primary health care, Aarhus County, Denmark. SUBJECTS: 181 patients with increased concentrations of plasma MMA ( > 0.28 micromol/l) attending 10 GPs. MAIN OUTCOME MEASURES: Reasons for requesting the analysis and reactions to a plasma MMA level above the reference interval. RESULTS: A reason for requesting plasma MMA was stated in 129 (71%) of the 181 medical records, screening being the most frequent. A reaction to an increased concentration of plasma MMA was recorded in 128 (71%) patients, and vitamin B-12 treatment was initiated in 102 (80%) of them. Among the 105 patients with marginally elevated plasma MMA level (0.29-0.44 micromol/l), vitamin B-12 treatment was initiated in 38 (36%). CONCLUSION: Plasma MMA was frequently used as a screening test. Treatment with vitamin B-12 was initiated when MMA level was increased, and often when plasma MMA level was slightly above the reference interval. This strategy is likely to promote over-treatment with vitamin B-12. PMID- 12086287 TI - Transcultural general practice in Scandinavia. AB - During the past 20 years, many immigrants from all over the world have settled in Scandinavia. Primary care physicians today are therefore meeting patients from a variety of socio-cultural, religious, and ethnic backgrounds. In addition to endemic diseases, the new immigrants are exposed to Western illness and disease patterns and psychic reactions to life events, and the on-going adaptive process may add to the dilemmas of segregation in housing and work. During consultations, doctors and patients frame this ill health by culturally determined ideas of health, illness, and treatment. This paper deals with Scandinavian studies concerning transcultural issues in primary care by exploring the Scandinavian literature. Relatively few studies were found in the databases. Many of them were small, making it difficult to generalise the findings. Descriptive explorative studies suggest problems in communication, behaviour, mental ill health, physiotherapy, and organisation of care. No studies were found concerning issues such as genital mutilation, environmental diseases, family conflicts, or chronic disorders other than pain. From action research studies or randomised-controlled trials, it can be cautiously concluded that psychiatric care may be little accepted in many immigrant groups and that immigrants with non-specific pain are best handled in primary care where dialogue about pain is to be preferred to traditional treatment. Brief advice regarding communication and organisation of care is also given. In our opinion, studies using action research methods are to be preferred, since clinical transcultural care deals with complex illness patterns, including many emotional dilemmas. PMID- 12086288 TI - Willingness of patients to perform self-management of asthma and the role of inhaled steroids. AB - OBJECTIVE: Asthma self-management is a generally accepted effective treatment strategy for asthma patients. Acceptance by patients may be a barrier for successful implementation. In this study, the role of inhaled steroids in starting asthma self-management is described. DESIGN: Cross-sectional explorative study. SETTING: General practice. SUBJECTS: 283 adult steroid-requiring asthma patients were invited by their GP to participate in a self-management programme. MAIN OUTCOME MEASURES: In a multivariate logistic regression model, the relation between baseline dosage of inhaled steroids, occupational status, age and sex as independent variables and willingness to participate as dependent variables was explored. RESULTS: Of all invited, 148 (52%) were willing to participate. Subjects not using inhaled steroids were least willing to participate (43/143 = 30%). Subjects with low doses of inhaled steroids ( < 400 mcg daily) were most willing to participate (44/54 = 81%). Unemployed asthmatics had a higher tendency to participate than patients with a regular job. CONCLUSION: Acceptance of self management by patients is not a limiting issue in promoting self-management of asthma in general practice. High acceptance in patients taking low or intermediate doses of inhaled steroids makes general practice the most appropriate setting for self-management. A selection procedure is recommended. PMID- 12086289 TI - Cyclooxygenase-2-selective inhibitors: translating pharmacology into clinical utility. AB - Anti-inflammatory agents have been used for centuries, but only in the last few decades has medical science gained insight into the complex biologic roles of the primary mediators of inflammation, the eicosanoids and their derivatives. Detailed understanding of the prostaglandins and leukotrienes provides a framework for the treatment of pain, inflammation, and fever with aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs), but these agents have exacted a substantial side effect burden. The discovery of cyclooxygenase-2 (COX 2) has guided development of rationally designed therapeutic agents that have the benefits of older NSAIDs with reduced gastrointestinal toxicity. Elucidation of the structure of COX isoenzymes has been key in the development of coxibs, the COX-2-selective subset of NSAIDs. Methods to determine the degree of COX-2 selectivity have been refined and are indispensable for comparing the relative selectivity of these agents. This review summarizes some of the key aspects of COX biochemistry, structure, and function and the evolution of understanding the mechanism of action of COX-2-selective inhibitors. The clinical relevance of COX 1 compared with COX-2 inhibition is discussed to provide a framework upon which clinicians can better appreciate current and future therapeutic applications of coxibs. PMID- 12086290 TI - COX-2-selective inhibitors in the treatment of arthritis. AB - Therapy with nonselective nonsteroidal anti-inflammatory drugs (NSAIDs) has long been the cornerstone of pharmacologic management of patients with osteoarthritis (OA) and rheumatoid arthritis (RA). Many patients with OA or RA, however, are at increased risk of developing clinically significant adverse events associated with NSAID therapy, particularly upper gastrointestinal (GI) complications including symptomatic and complicated ulcers. The introduction of cyclooxygenase (COX)-2-selective inhibitors (coxibs) represents a major advance in the pharmacologic approach to the signs and symptoms of arthritis. In addition to the first two members of this class, celecoxib and rofecoxib, other coxibs have been introduced or are in development (valdecoxib, etoricoxib). In numerous clinical trials, coxibs have been shown to be as effective as nonselective NSAIDs in relieving pain and inflammation associated with OA and RA, and notably, with a significantly lower risk of NSAID-type adverse events. The use of coxibs to treat OA and RA is recommended as first-line therapy when symptoms of pain and inflammation are present in patients vulnerable to potential NSAID-associated GI toxicity. PMID- 12086291 TI - Gastrointestinal safety and tolerability of nonselective nonsteroidal anti inflammatory agents and cyclooxygenase-2-selective inhibitors. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most widely used of all drugs and are the most common medications used by persons aged 65 years or more. NSAIDs have a number of side effects, of which the most prevalent and serious is gastrointestinal (GI) toxicity. GI side effects of NSAIDs range from dyspepsia and gastroduodenal ulcers to serious, potentially fatal GI complications including bleeding and perforation. Serious GI complications often lack warning signs; knowledge of risk factors for NSAID-related gastropathy can identify patients at high risk, allowing for initiation of the appropriate therapeutic intervention. Risk factors include advanced age, NSAID dose, prior GI complications, infection with Helicobacter pylori, and use of corticosteroids and anticoagulants. There are few well-established strategies to prevent GI complications in NSAID users. Risk assessment and cotherapy with acid suppressors (H2-receptor antagonists and proton pump inhibitors) or prostaglandin replacement (misoprostol) and H pylori eradication are beneficial. Cyclooxygenase-1 (COX-1) is a key enzyme in gastroprotective mucosal defenses, and the best way to prevent GI toxicity is to avoid drugs that inhibit COX-1. Clinical studies of the COX-2 selective inhibitors rofecoxib and celecoxib have demonstrated efficacy equivalent to nonselective NSAIDs with lower rates of GI side effects (for example, incidence of endoscopic ulcers equivalent to placebo). Selective COX-2 inhibitors (coxibs) provide effective treatment of pain and inflammation while reducing risk of gastropathy. PMID- 12086292 TI - Outcomes studies of the gastrointestinal safety of cyclooxygenase-2 inhibitors. AB - Short-term endoscopic studies of the highly selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) rofecoxib and celecoxib have shown that these agents are well tolerated and have efficacy equivalent to nonselective nonsteroidal anti inflammatory drugs (NSAIDs) with fewer adverse effects on the upper gastrointestinal (GI) tract. These studies are limited, however, as the detection of endoscopic lesions is not well correlated with symptomatic ulcers and ulcer complications. Outcomes studies of the GI safety are, therefore, essential to understanding how coxibs are likely to perform in a clinical practice setting. Four large outcomes studies (Vioxx Gastrointestinal Outcomes Research, VIGOR; Assessment of Difference Between Vioxx and Naproxen to Ascertain Gastrointestinal Tolerability and Effectiveness trial, ADVANTAGE; Celecoxib Long-term Arthritis Safety Study, CLASS; and the Successive Celecoxib Efficacy and Safety Studies, SUCCESS) examined the GI safety of rofecoxib and celecoxib in over 39,000 patients with osteoarthritis or rheumatoid arthritis. Results of these studies showed that patients taking a supratherapeutic dose of rofecoxib or celecoxib had significantly lower rates of GI-related adverse events than those taking a nonselective NSAID (naproxen, ibuprofen, or diclofenac). Reduced risk of upper GI events was seen in patients with multiple risk factors and in patients using low dose aspirin and corticosteroids concomitantly with a coxib. Results of large outcomes studies provide support for the COX-2 hypothesis and demonstrate the long-term safety and tolerability of coxibs. PMID- 12086294 TI - Development and clinical application of COX-2-selective inhibitors for the treatment of osteoarthritis and rheumatoid arthritis. AB - Osteoarthritis (OA) and rheumatoid arthritis (RA) are among the most prevalent chronic illnesses and leading causes of disability in the United States. The clinical symptoms of OA and RA, pain and inflammation, are biologic processes mediated in part by prostanoids-prostaglandins, prostacyclin, and thromboxanes. The intermediate enzymes responsible for prostaglandin biosynthesis, cyclooxygenase (COX)-1 and COX-2, have been the target of arthritis therapy using nonselective nonsteroidal anti-inflammatory drugs (NSAIDs). An understanding of the biochemistry and molecular pharmacology of COX enzymes has allowed for the development of agents that specifically inhibit COX-2. COX-2-selective inhibitors have efficacy in OA and RA that is similar to that of NSAIDs but with a lower potential for upper gastrointestinal injury, a serious side effect of nonselective NSAIDs. COX-2-selective inhibitors have been increasingly used in the treatment of OA and RA as well as other inflammatory arthropathies including ankylosing spondylitis and gout. Clinical trials with two currently available drugs, rofecoxib and celecoxib, have demonstrated efficacy comparable to nonselective NSAIDs but with a lower risk of gastrointestinal side effects. In general, these drugs are well tolerated in patients with aspirin-sensitive asthma. Rofecoxib is well tolerated in patients with sulfonamide sensitivities; further studies are needed to fully characterize the utility of celecoxib in these patients. Clinical experience shows that because of their improved GI safety, rofecoxib and celecoxib, and newer COX-2-selective inhibitors (valdecoxib, etoricoxib, parecoxib), represent a significant advance in the treatment of arthritis and other related inflammatory conditions. PMID- 12086293 TI - Current perspective on the cardiovascular effects of coxibs. AB - Aspirin and nonselective nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used for their anti-inflammatory and analgesic effects. In addition, aspirin is documented to reduce cardiovascular events in selected populations, presumably because of inhibition of platelet aggregation. Yet these drugs are not without toxicity, particularly adverse effects on the gastric mucosa. The gastrointestinal toxicity of nonselective NSAIDs and aspirin derives from the inhibition of the cyclooxygenase (COX) enzyme, COX-1, which synthesizes gastroprotective prostaglandins, while the anti-inflammatory and pain-relieving effects are largely derived from inhibition of COX-2-derived prostaglandins. Available data indicate that the harmful gastric effects of nonselective NSAIDs are reduced by substitution of agents that only inhibit the COX-2 protein. The COX-2-selective inhibitors, however, have also been shown to inhibit the production of vascular prostacyclin, which has vasodilatory effects and inhibits platelet aggregation; unlike nonselective NSAIDs, they do not inhibit the production of thromboxane, an eicosanoid that promotes platelet aggregation. Whether these effects could potentially contribute to a prothrombotic environment is the subject of current, intensive debate. In the Vioxx Gastrointestinal Outcomes Research (VIGOR) trial, there was a higher incidence of cardiovascular thrombotic events in the rofecoxib- vs the naproxen-treated group: 1.67 vs 0.70 per 100 patient years. However, in a pooled analysis of rofecoxib studies, the risk of sustaining a thrombotic cardiovascular event was similar when comparing patients receiving rofecoxib with those receiving placebo, or when comparing patients receiving rofecoxib with those receiving nonnaproxen nonselective NSAIDs. These findings are likely to result, at least in part, from the antiplatelet action of naproxen, which has been shown to be potent and sustained during a typical dosing regimen (500 mg twice daily in VIGOR). In contrast, the other NSAID comparators effect weaker and/or nonsustained antiplatelet action. In the Celecoxib Long-term Arthritis Safety Study (CLASS) trial, there was no difference between celecoxib and the nonselective NSAIDs explored (which did not include naproxen) in cardiovascular event rates. Unlike those in VIGOR, patients in the CLASS trial were allowed to take low-dose aspirin. Thus, despite concerns raised by results of VIGOR, other existing data, including those pooled from existing placebo-controlled trials, do not support a clinically relevant prothrombotic effect of the COX-2 inhibitors. Additional placebo-controlled data, from patients at both high and low risk for cardiovascular events, are warranted to clarify the cardiovascular effects of this class of agents. PMID- 12086295 TI - Renal effects of nonselective NSAIDs and coxibs. AB - Despite the ubiquitous use of both over-the-counter and prescription nonsteroidal anti-inflammatory drugs (NSAIDs), clinical syndromes-NSAID-related hypertension, salt and water retention, edema, and hyperkalemia-are highly infrequent. Nevertheless, they remain a concern, and patient populations at risk for renal adverse effects from NSAIDs can be prospectively identified. Patients at risk include those with age-related declines in glomerular filtration rate; those with hypovolemia, particularly patients taking loop diuretics; and those with congestive heart failure, cirrhosis, or nephrosis. The following patient populations are at higher risk for increases in blood pressure with concomitant use of an NSAID and an antihypertensive: those with congestive heart failure, liver disease, or kidney disease, and those taking angiotensin-converting enzyme inhibitors or diuretics. Nonselective NSAIDs and COX (cyclooxygenase)-2-selective inhibitors (coxibs) appear to have similar effects on renal function if dosed equivalently, and standard precautions to avoid renal toxicity with use of nonselective NSAIDs apply to coxibs. PMID- 12086296 TI - Developing an economic rationale for the use of selective COX-2 inhibitors for patients at risk for NSAID gastropathy. AB - Arthritis causes considerable patient morbidity and substantial health care resource utilization. One important contributing component to the overall cost burden of this condition is the variety of expenditures attributable to the adverse effects of arthritis therapy. Nonselective nonsteroidal anti-inflammatory drugs (NSAIDs) are a mainstay of medical treatment for patients with arthritis because of their well-established anti-inflammatory and analgesic effects. Generally well tolerated, traditional NSAIDs nevertheless cause adverse gastrointestinal (GI) effects in a proportion of patients. Because nonselective NSAIDs are so widely used, these GI adverse events cause significant morbidity and mortality, accounting for substantial additional health care expenditures. Data from controlled investigations document the enhanced GI safety of cyclooxygenase (COX)-2-selective inhibitors, or coxibs, when compared with nonselective NSAIDs. As a result of this improved safety profile, patients treated with coxibs use significantly fewer GI-related health care resources (eg, medications, procedures) than patients treated with nonselective NSAIDs. Thus, available clinical and economic data suggest that the use of coxibs has the potential to result in important clinical GI benefits at an acceptable incremental cost for all chronic NSAID users. For individuals who are at an increased risk of developing GI complications attributable to NSAIDs, coxibs are clearly a cost-effective treatment option. PMID- 12086297 TI - Cyclooxygenase-2-selective inhibitors in the management of acute and perioperative pain. AB - Postsurgical pain is often undertreated. Opioids are frequently used in perioperative analgesia, but concern about side effects can result in administration of an inadequate dose for pain relief. Nonsteroidal anti inflammatory drugs (NSAIDs) are used increasingly for postoperative analgesia. The use of balanced analgesia-a combination of opioids, NSAIDs, and local anesthesia utilizing agents from other classes (eg, ketamine, clonidine)-improves the efficacy of pain relief and decreases risk of side effects. While lacking some of the troublesome side effects of opioids, nonselective NSAIDs may cause bleeding as a result of their inhibitory effects on COX-1. For this reason, COX-2 selective inhibitors (coxibs) are attractive opioid-sparing analgesic options in the perioperative setting. Factors in addition to side effects such as time to onset of action, duration of action, maximum pain relief, use of rescue medication, and other factors relevant to a given pain model are important in determining overall analgesic efficacy. Clinical studies show that COX-2 selective inhibitors are effective for the treatment of preoperative and postoperative pain and reduce postsurgical requirements for opioids. This evidence supports a role for COX-2-derived prostaglandins as key mediators of nociceptive pain and peripheral sensitization (hyperalgesia). Pain management in the perioperative setting and the role of COX-2-selective inhibitors in acute and postoperative pain are reviewed here. PMID- 12086299 TI - My patient, my subject, my patient. PMID- 12086298 TI - Emerging options with coxib therapy. AB - Future clinical applications of cyclooxygenase (COX)-2-selective inhibitors (coxibs) are likely to extend beyond their current use as oral analgesics in high risk arthritis patients. The clinical utility of coxibs for the treatment of Alzheimer's disease (AD) is under investigation. Epidemiological surveys, preclinical studies, and preliminary clinical trials with nonsteroidal anti inflammatory drugs (NSAIDs) have suggested that inflammatory mechanisms play a role in the neurodegeneration of AD. Clinical trials are currently being conducted to determine the effect of coxibs on the rate of AD progression. The use of coxibs as chemopreventive agents in colorectal cancer (CRC) is also under investigation. The chemopreventive benefits of coxibs to promote cell death (apoptosis) and inhibit angiogenesis in CRC have been shown in tumor cell lines and in animal and human models. In addition, palliative care clinicians and oncologists are increasingly including coxibs in their management of cancer pain. Coxibs are utilized for their opioid-sparing effect in the management of cancer pain, without impairing wound healing, or promoting bleeding diathesis (antiplatelet effects) or adverse gastrointestinal effects in patients receiving chemotherapy or radiation treatment. PMID- 12086300 TI - Juxtapapillary capillary hemangioblastoma. AB - BACKGROUND: Juxtapapillary capillary hemangioblastomas are vascular lesions that occur on the optic nerve head or immediately adjacent to the optic disc. Although juxtapapillary capillary hemangioblastomas may appear as an isolated clinical finding, they are more likely to be a precursor to the diagnosis of von Hippel Lindau (VHL) disease. When associated with VHL, the ocular complications from hemangioblastoma are generally greater, and subsequently the prognosis is poorer than for isolated entities. CASE REPORT: A 67-year-old white male presented to our clinic complaining of decreased vision in both eyes at distance and near. A dilated fundus examination revealed a swollen left optic nerve. Visual field testing revealed an inferior arcuate defect in the left eye. Laboratory and radiological testing were normal. Fluorescein angiography showed filling during the retinal arterial phase with late diffuse leakage. Based on these results, a diagnosis of juxtapapillary capillary hemangioblastoma was made. This was later confirmed when fundus photographs taken 7 years earlier were obtained, revealing a stable appearance to the nerve. CONCLUSION: Potential ocular complications of juxtapapillary capillary hemangioblastomas include vision loss secondary to subretinal and intraretinal fluid, as well as epi-retinal membrane formation and, rarely, exudative retinal detachment. Management ranges from observation to surgical intervention, although definitive treatment guidelines have yet to be established. In addition, it is important for the eye care practitioner to be aware of the association with VHL disease, so that patients may be appropriately referred and managed for other possible systemic complications of the disease. PMID- 12086301 TI - Ocular and systemic findings and their correlation with hemodynamically significant carotid artery stenosis: a retrospective study. AB - BACKGROUND: Optometrists often encounter patients with ocular signs and/or symptoms suggestive of carotid artery disease, but criteria for eye care practitioners concerning when to order carotid studies are not well established. A retrospective study in an optometry clinic was performed to determine if certain ocular findings and associated systemic risk factors were associated with hemodynamically significant carotid artery stenosis (HSCAS). METHODS: A retrospective analysis was performed on all patients examined in the optometry clinic at the Sepulveda Ambulatory Care Center from January 1, 1998 through December 31, 1999 to identify all patients who had carotid studies ordered. Charts were then reviewed to determine the ocular finding that prompted the carotid study. Statistical analysis using an odds-ratio was performed to determine whether any ocular sign/symptom or systemic risk factor was associated with HSCAS. RESULTS: Of 3822 patients, 48 (1.26%) had carotid studies ordered. Eight (17%) had HSCAS of >50% and symptomatic patients were 1.6 times more likely to have HSCAS than asymptomatic patients. Patients with HSCAS were 1.8 times more likely to have retinal vascular occlusions, 1.9 times more likely to have normotensive glaucoma, 2.4 times more likely to have peripheral retinal hemorrhages, and 2.6 times more likely to be smokers, although none of these factors were found to be a statistically significant indicator of HSCAS. However, the number of systemic diseases tended to be greater for the HSCAS patients compared with the non-HSCAS patients (HSCAS median = 3.5 and the non-HSCAS median = 2, p = 0.049). CONCLUSION: Although no single ocular or systemic risk factor was found to be a statistically significant indicator of HSCAS, the number of systemic disease risk factors tended to be greater for the HSCAS patients compared with the non-HSCAS patients (p = 0.049). An extensive literature review was also performed to help establish guidelines for when to order carotid studies on the basis of ocular signs and/or symptoms as well as the presence of additive associated systemic risk factors. PMID- 12086302 TI - Evaluation of tear film interference patterns and measures of tear break-up time. AB - PURPOSE: The purpose of this study was to compare the agreement within and between examiners when evaluating real-time and digital photographs of TearScope Plus tear film interference patterns. We also evaluated between-examiner agreement of noninvasive digital tear break-up time (NITBUT) using the TearScope, the relation between NITBUT and tear film interference patterns, and the relation between NITBUT and tear break-up time (TBUT). METHODS: Forty nondry eye patients were seen for digital imaging of the tear film using the TearScope Plus mounted on a slit-lamp with the EyeCap Ophthalmic Image Capture System. Two independent examiners first assessed the tear interference pattern in real-time and then captured the patterns and NITBUT digitally for a period of approximately 60 s. Each image series was later graded for interference pattern and NITBUT by the two independent, masked examiners. RESULTS: Within-examiner comparisons of real-time and digital tear pattern photograph grading showed moderate to substantial agreement for experienced examiners (kappa for examiner 1 = 0.76 and kappa for examiner 2 = 0.55). Between-examiner agreement when grading tear patterns was also substantial in real-time (kappa = 0.72) and moderate when using digital photography (kappa = 0.59). The between-examiner mean NITBUT was 11.2+/-6.8 s and the 95% limits of agreement between examiners were -19.2 to +9.2 s. The mean TBUT was 7.6+/-10.4 s, and it was on average 3.7+/-12.0 s shorter than NITBUT (p = 0.06). Thicker tear film interference patterns graded photographically were associated with longer NITBUTs for both examiners (p = 0.001). CONCLUSIONS: There is moderate to substantial within- and between-examiner agreement when comparing real-time and digital tear interference patterns photographs when using the TearScope. Although there is considerable between-examiner variability with the NITBUT test, the thicker lipid layer tear patterns tend to be associated with longer NITBUTs. PMID- 12086303 TI - Interexaminer repeatability of a new, modified prentice card compared with established phoria tests. AB - PURPOSE: The Howell phoria card, a new modification of the Prentice design, has no previously published validity or reliability data. The purpose of this study was to compare the interexaminer repeatability of the Howell phoria card with established tests. METHOD: Experienced optometrists measured the near heterophoria of 72 subjects. Heterophoria was measured in a pseudo-random order using five standardized procedures: (1) Howell phoria card continuous presentation, (2) Howell phoria card flashed presentation, (3) free-space von Graefe method, 4) Bernell Muscle Imbalance Measure (MIM) card continuous presentation, and (5) MIM card flashed presentation. RESULTS: The von Graefe method and the Howell phoria card flashed presentation found significantly less exophoric results than the MIM card. There was no consistent tendency for different examiners to find more exo- or more esophoric results with the phoria measurement techniques used in this study (p = 0.28). Both the Howell phoria card continuous presentation and the MIM card had significantly less variability in interexaminer differences than the Howell phoria card flashed presentation and the von Graefe method (p < 0.05). CONCLUSIONS: Results from this study suggest that the Howell phoria card used in a continuous presentation method has an interexaminer repeatability that is not significantly different from the MIM card but greater than the von Graefe method. PMID- 12086304 TI - The effectiveness of occluder contact lenses in improving occlusion compliance in patients that have failed traditional occlusion therapy. AB - PURPOSE: Management of infants and young children with dense amblyopia, including that which results from unilateral congenital cataracts, is challenging because of noncompliance with occlusion therapy. Occluder contact lenses (OCLs) have been described to successfully improve visual acuity in patients with amblyopia. The purpose of this study was to evaluate the effectiveness of OCLs in improving occlusion compliance in infants and young children with dense amblyopia who had failed traditional occlusion therapy. METHODS: Thirteen patients were fit in their nonamblyopic eye with OCLs provided by Wesley Jessen (Des Plaines, IL). All patients had dense amblyopia as diagnosed by a pediatric ophthalmologist. Nine of the 13 patients had deprivational amblyopia associated with unilateral aphakia, 2 patients had strabismic amblyopia, and the other 2 had mixed mechanism amblyopia. All patients had failed traditional occlusion methods. Occlusion was prescribed on a part-time basis to prevent occlusion amblyopia to the better-seeing eye. Compliance with the OCLs was determined by questioning the parents as to whether the prescribed occlusion time was accomplished. RESULTS: Five of the 13 patients (38.4%) did not succeed in the initial 1-mo trial period because of parental difficulties with lens handling. Four patients (30.7%) achieved partial success, as they were able to wear the lens between 4 to 14 mo, at which time they learned to remove the occluder lens. The four remaining patients (30.7%) successfully wore the occluder lenses until the conclusion of the study, a time period ranging from 26 to 60 mo. No complications resulted from OCLs. CONCLUSIONS: OCLs can provide an alternative to achieving occlusion compliance after the failure of traditional occlusion therapy in infants and young children with dense amblyopia. PMID- 12086305 TI - Contact lens drying and visual performance: the vision cycle with contact lenses. AB - BACKGROUND AND PURPOSE: The purpose of this study was to measure the effect of precontact lens tear film break-up on visual performance. METHODS: Four asymptomatic soft contact lens wearers had contrast sensitivity measured by a temporal, two-alternative, force choice paradigm combined with a self-paced methods of limits. Stimuli were vertically orientated sine wave gratings (0.5 to 14 cycles per degree [cpd] presented for 16.67 ms. Contrast sensitivity was measured before precontact lens tear break-up by a stimuli presented 2 s after the blink. A post-tear layer break-up measurement taken with the stimuli presented after break-up had been observed by the use of a video camera attached to a Tearscope. RESULTS: Contrast sensitivity was found to be reduced following precontact lens tear film break-up for stimuli of 4, 6, and 10 cpd; the data approached significance at 14 cpd. Further reductions in contrast sensitivity were observed for one subject when measurements were continued for 4 s following break-up. CONCLUSIONS: Contrast sensitivity is significantly reduced for middle to high spatial frequencies when the precontact lens tear film dries and breaks up. The combination of observations of visual performance immediately following the blink (from earlier experiments) and measurements following tear film break up in this experiment allows description of a "vision cycle" for contact lens wearers in the interval between blinks. It is suggested that break-up of the precontact lens tear film could account for the complaints of intermittent blurred vision in some contact lens wearers and may provide a stimulus to blinking in these individuals. PMID- 12086306 TI - The effects of optical defocus on the legibility of the Tumbling-E and Landolt-C. AB - BACKGROUND: The Tumbling-E and Landolt-C are both used to test visual acuity in young, illiterate, or non-English-speaking patients. In this study, we sought to determine how optical blur affects visual acuity thresholds measured with both of these optotypes. METHODS: Ten subjects were tested with 4-position Tumbling-E and Landolt-Cs using the method of constant stimuli. The subjects were tested wearing their habitual corrections and with +1.50 D spherical and cylindrical lenses to simulate myopic refractive errors in the optical blur test conditions. RESULTS: Visual acuity thresholds for the Tumbling-E were slightly better for all test conditions, while the largest threshold difference between the test letters was found for the simulated against-the-rule astigmatism. For both test letters, vertical orientations were more difficult to resolve than horizontal. Also, the slopes of the psychometric functions were steeper for the Tumbling-E than for the Landolt-C. CONCLUSIONS: The Tumbling-E can be considered comparable with the Landolt-C except for the simulated against-the-rule-astigmatism test condition. Although the ranges are quite large, the average psychometric functions of the Landolt-Cs are flatter, which gives rise to the possibility that more confidence can be placed in the visual acuity thresholds obtained with the Tumbling-E. PMID- 12086307 TI - Monoclonal antibodies against rat leukocyte surface antigens. AB - Monoclonal antibodies have proven to be powerful tools for studying the properties of leukocyte surface antigens and the cells that express them. In the past decades many monoclonal antibodies (mAb) for identifying the different rat leukocyte surface antigens have been described. A list of mAb is provided in Table I below. The rat leukocyte surface antigens are divided into different sections, including rat CD antigens (a), rat leukocyte surface antigens without CD designation (b), rat major histocompatibility complex (MHC) antigens (c), rat T-cell receptors (d) and rat immunoglobulins (e). The molecular and functional characteristics of rat leukocyte surface antigens are discussed in more detail in some of the other chapters of this issue (e.g. Van den Berg et al., p. 45). A more extensive overview of the properties of leukocyte surface antigens is provided by Barclay et al. PMID- 12086308 TI - Experimental autoimmune encephalomyelitis in the rat: lessons in T-cell immunology and autoreactivity. AB - Experimental autoimmune encephalomyelitis (EAE) in the rat is an acute paralytic disease from which most animals spontaneously recover. The disease can be induced in susceptible inbred Lewis and DA rats with myelin basic protein (MBP), or encephalitogenic MBP peptides administered in complete Freund's adjuvant (CFA). The disease can be adoptively transferred to syngeneic recipients with primed T cells that have been reactivated in vitro with antigen. EAE is mediated by CD4+ Th1 cells that secrete proinflammatory cytokines, and spontaneous remission is associated with CD4+ T cells that secrete transforming growth factor-beta (TGF beta). Studies of EAE in susceptible rats have provided many important insights into the interactions of T cells and accessory cells that culminate in the induction of the autoimmune response. PMID- 12086309 TI - Cellular and genetic factors involved in the difference between Brown Norway and Lewis rats to develop respectively type-2 and type-1 immune-mediated diseases. AB - The understanding of the mechanisms of immune tolerance and the unravelling of the pathophysiology of autoimmune diseases rely on animal models. In this respect, BN and LEW rats represent models of choice to study immune-mediated diseases from the cellular and genetic points of view. Indeed, BN and LEW rats are extremes with respect to their polarisation of the immune response as well as their susceptibility to autoimmune diseases. LEW rats are susceptible to Th1 mediated autoimmune diseases while BN rats are highly susceptible to Th2-mediated autoimmune disease. Comparison of the T cell compartment between LEW and BN rats revealed several important differences. 1) A MHC-dependent quantitative difference that is due to a defect in the CD8 T cell compartment in BN rats. 2) A qualitative MHC-independent difference that is related to a high frequency of CD45RClow CD4 and CD8 T cell subsets, producing IL-4, IL-13, IL-10 and TGF-beta in BN rats as compared to LEW rats. 3) Interestingly, the genetic studies showed that susceptibility to Th1-mediated experimental autoimmune encephalomyelitis, and to Th2-mediated disorders triggered by gold salts as well as the difference in the CD4SRChigh/CD45RClow ratio between LEW and BN rats are genetically determined by regions on chromosomes 9, 10 and 20. PMID- 12086310 TI - BB rat lyp mutation and Type 1 diabetes. AB - BioBreeding (BB) rats spontaneously develop an autoimmune diabetic syndrome similar to that observed in humans and NOD mice. One of the diabetes susceptibility loci maps to the lyp locus on chromosome 4. In this article we describe the consequences of the BB rat lyp mutation on T-cell homeostasis, repertoire and function, as well as its role in the pathogenesis of type I diabetes. PMID- 12086311 TI - Genetic analysis of collagen-induced arthritis in rats: a polygenic model for rheumatoid arthritis predicts a common framework of cross-species inflammatory/autoimmune disease loci. AB - Collagen-induced arthritis (CIA) is a useful model for dissecting the genetic patterns underlying susceptibility to rheumatoid arthritis (RA) and related chronic/inflammatory autoimmune diseases. CIA exhibits three phenotypes characteristic of autoimmune disease pathogenesis: abnormal levels of immune reactivity to self antigens; chronic inflammation of target organs expressing that specific autoantigen; activation and direct participation of invading mononuclear cells and resident tissue fibroblasts in organ damage. Over 25 different quantitative trait loci (QTL) regulating arthritis severity and autoantibody in rats with CIA are mapped. QTL-congenic strains show that certain CIA-QTLs can modulate arthritis independently These monogenic models are proving to be highly informative for fine mapping and function studies, revealing gender effects and evidence of gene clusters. Recent genome scans of RA populations identified RA-susceptibility loci in chromosome regions homologous to rat chromosomal segments housing CIA-QTLs. Also, CIA-QTLs frequently co-localize with susceptibility QTLs mapped in other rat arthritis models induced with non immunogenic adjuvant oils and/or in rat autoimmune models of multiple sclerosis and diabetes. Common autoimmunity genes and inflammation genes important to several human diseases are likely being detected in the various rat disease models. Continued dissection of the genetic underpinnings of rat arthritis models should provide candidate genes for investigation in human patients and lead to a clearer understanding of the complex genetics of RA. PMID- 12086312 TI - Arthritis induced in rats with nonimmunogenic adjuvants as models for rheumatoid arthritis. AB - Rat models are useful for studies of the pathogenesis of rheumatoid arthritis (RA) since rats are extraordinarily sensitive to induction of arthritis with adjuvants. Injection of not only the classical complete Freund's adjuvant but also mineral oil without mycobacteria and pure adjuvants such as pristane and squalene, induce severe arthritis in many rat strains. Models like pristane induced arthritis in rats are optimal models for RA since they fulfill the RA criteria including a chronic relapsing disease course. Arthritogenic adjuvants like pristane, avridine, squalene and mineral oil are not immunogenic since they do not contain major histocompatibility complex (MHC) binding peptides. Nevertheless, the diseases are MHC-associated and dependent on the activation of alphabetaTCR (T-cell receptor)-expressing T cells. However, it has not been possible to link the immune response to joint antigens or other endogenous components although immunization with various cartilage proteins induce arthritis but with different pathogeneses. To unravel the mechanisms behind adjuvant induced arthritis, a disease-oriented genetic approach is optimal. Several loci that control onset of arthritis, severity and chronicity of the disease have been identified in genetic crosses and most of these have been confirmed in congenic strains. In addition, many of these loci are found in other autoimmune models in the rat as well as associated with arthritis in mice and humans. PMID- 12086313 TI - Migration of naive, effector and memory T cells: implications for the regulation of immune responses. AB - T cells play an important role in protective immune responses and in the pathogenesis of many diseases. Understanding the mechanisms regulating their distribution in vivo may therefore be of therapeutic value. Reviewing studies that have followed the migration of labelled naive, effector and memory T cells in healthy animals reveals that all T-cell subsets enter all organs investigated. Within the tissue, two principally different migration patterns can be identified. First, naive and memory T cells accumulate in lymphoid organs for about 48 h after injection, as the time needed for migration through lymphoid organs is longer than through non-lymphoid organs. During this time, surface molecule expression is temporarily modified. These changes are reversed before leaving the lymphoid organs and entering the blood to start a new cycle of migration. Second, effector T cells are evenly distributed throughout the body, and most die in the tissues within 24 h. However, depending on the presence of cytokines, some are able to survive and to proliferate, and thereby accumulate in defined microenvironments of the body. Analysing the principles regulating T-cell migration and survival within the tissue may lead to the development of new options for the treatment of disease. PMID- 12086314 TI - Neuroinflammation in the rat--CNS cells and their role in the regulation of immune reactions. AB - Recent discoveries suggest that the resident cells of the central nervous system (CNS) the nerve cells and glia, play a more immunologically active role than was previously assumed. Neuroglial communication is of central interest in virtually all types of pathological conditions that affect the brain and several features of the activation that results from nerve cell damage resemble the type of innate immune reactions that occur in other parts of the body In particular, the characteristics of the activation of these CNS cells will affect both the interaction with cells of the immune system as well as processes related to neurodegeneration and regeneration. We here review data regarding 3 different aspects of local inflammatory activation in the rat nervous system: (i) the genetic heterogeneity of glial activation across inbred strains after nerve injury, (ii) expression of MHC class I genes in the CNS and (iii) neuroprotective effects of CNS antigen autoreactive immune reactions. Apart from neuroimmune diseases such as experimental autoimmune encephalomyelitis/multiple sclerosis, these features are also of relevance for a wider range of neurological diseases which present pathological signs of inflammation, such as Alzheimer's dementia, cerebrovascular diseases and CNS trauma. PMID- 12086315 TI - Conditioning in the rat: an in vivo model to investigate the molecular mechanisms and clinical implications of brain-immune communication. AB - A wealth of in vitro and ex vivo evidence has described both close anatomical interaction and functional bi-directional communication between the immune and central nervous systems (CNS). These data have provided a framework for understanding the physiological mechanisms whereby behavioural factors may impact immune-related disease. An understanding of this interaction, however, as well as verification of the biological relevance of communication among these systems, requires in vivo animal modelling. The development of psychoneuroimmunological models in the laboratory rat has played a key role in advancing the understanding of the influence of behaviour on immune status. One such paradigm is the behavioural conditioning of immune function in the rat. This elegant model is characterised by the ability to examine simultaneously both afferent and efferent brain-immune communication. Specifically, the role of peripheral cytokines in signalling the brain, as well as their anatomical and cellular targets in the CNS, can be identified. On the other hand, the neural and humoral pathways whereby the CNS influences the function and distribution of peripheral immunocytes can be demonstrated, together with the target hormone receptors on immunocompetent cells. Finally, the in vivo biological relevance of brain-immune communication is revealed by behavioural conditioning, demonstrating that clinically relevant conditions such as heart allograft survival can be modified by behavioural processes. Behavioural conditioning thereby provides an excellent example of the utility of in vivo laboratory rat models in psychoneuroimmunology research. Such paradigms not only provide a more complete knowledge of CNS-immune system interaction, but are the platform for determining potential clinical application of this information. PMID- 12086316 TI - Role of NK cells in adoptive immunotherapy of metastatic colorectal cancer in a syngeneic rat model. AB - This article reviews our immunotherapy research with natural killer (NK) cells in a syngeneic rat colorectal cancer liver and lung metastasis model. Using adoptive transfer of interleukin (IL)-2-activated NK cells, NK cells were shown to selectively infiltrate the tumors. More NK cells were found in tumors when the NK cells were directly injected into tumor-draining blood vessels than when the cells were injected in systemic blood vessels. Under optimal conditions, a limited, though significant, effect of adoptively transferred NK cells on tumor growth was shown. We observed that both endogenous and adoptively transferred NK cells were predominantly present in the stroma surrounding the tumor cell nodules. It is possible that they did not penetrate the nodules containing the tumor cells because of the presence of a basal membrane-like structure around these nodules. Adoptively transferred NK cells may initiate elimination of tumor cells by activating other effector cells, whereas some may eliminate tumor cells by direct cell-cell contact. A diverse array of molecules was shown to be involved in this process. CD45 on NK cells was found to be important in initiating the lysis-inhibitory signal upon binding of 'self' major histocompatibility complex (MHC) class I on potential target cells. Our results indicate that NK-cell cancer therapy is still promising and needs improvement. PMID- 12086317 TI - Host-tumor interactions during the regression of a rat histiocytoma, AK-5. AB - An efficient immune response comprises a highly intricate, integrated circuitry involving both the cellular and the humoral arms of the immune system of the host interacting with the rapidly proliferating microcosm of the tumor The mechanism of tumor rejection involving multiple arms of the immune system was reviewed in a spontaneously regressing rat histiocytoma, AK-5, an autologous tumor-host system. Intraperitoneal tumor transplantation leads to death in all animals, whereas subcutaneously (s.c.) transplanted tumor undergoes regression in 70% of animals. Regression of the tumor occurs by both apoptosis and necrosis, and natural killer (NK) cells were identified as the chief effectors mediating tumor cell death in vivo. A type 1 helper T cell (Th1)-driven cytokine cascade played a crucial role in enhancing cellular functions at the tumor site and obtaining a sufficient immune response for tumor rejection. The s.c. tumor-bearing hosts were shown to produce a factor which induced apoptosis in tumor cells, mediating tumor rejection. This review emphasizes the daunting complexities and interesting liaisons between the host immune system and the tumor, highlighting the work from our laboratory, and stressing that it is the interaction of several factors in concert or antagonizing each other that is responsible for the spontaneous regression of a tumor. PMID- 12086318 TI - Heme oxygenase-1, a protective gene that prevents the rejection of transplanted organs. AB - Endothelial cells (EC) play a pivotal role in regulating inflammatory reactions such as those involved in the rejection of transplanted organs. This occurs through the expression of a series of pro- and anti-inflammatory genes that are associated with the activation of these cells. Presumably, the expression of pro inflammatory genes promotes events that lead to graft rejection, while expression of anti-inflammatory (protective) genes suppresses those events and thus contributes in sustaining graft survival. Understanding how the expression of these genes is regulated and their mechanism of action are important issues for the development of new therapeutic strategies to suppress graft rejection. We have studied this phenomenon using experimental models of transplantation in rats. We discuss here data that supports the concept that grafts can express anti inflammatory (protective) genes that mitigate inflammatory reactions leading to graft rejection. The data reviewed focus on the role of one of such genes, the stress responsive gene heme oxygenase-1, and of its byproduct carbon monoxide, which can suppress graft rejection and lead to long-term graft survival. PMID- 12086319 TI - DNA methylation and Z-DNA formation as mediators of quantitative differences in the expression of alleles. AB - With draft copies of several model genomes available in the near future, attention is turning towards the genetic mechanisms that determine differences between individuals. While mutations in protein coding regions affect the structure of gene products, polymorphisms outside such regions may cause quantitative differences in gene expression. Here we summarize observations indicating that such differences may be mediated by allele-specific alterations in the modification or structure of DNA. Mono-allelic expression of the rat T cell differentiation marker RT6 in a subpopulation of cells is associated with allele-specific differences in DNA methylation in the RT6 promoter. In contrast to previously described examples of mono-allelic expression, these are determined neither stochastically nor by parental origin, but by cis-acting elements within the alleles. An attractive candidate is a rodent identifier (ID) element exclusively present in the RT6a allele. In the case of the rat nucleolin gene, a polymorphic dinucleotide repeat in the 5' region modulates promoter strength and forms left-handed Z-DNA in vivo. Models explaining putative effects of Z-DNA formation on transcription are presented. These observations suggest novel mechanisms by which repetitive DNA, an abundant source of polymorphism in the mammalian genome, may exert quantitative effects on gene expression. PMID- 12086320 TI - Monocytes in the rat: phenotype and function during acute allograft rejection. AB - Cells of the monocyte/macrophage system originate from the bone marrow, reach the organs via the blood, immigrate through postcapillary venules and further differentiate into organ-specific tissue macrophages. In rats and other species, activated monocytes/macrophages aggravate autoimmune reactions, rejection of non vascularized allografts and chronic allograft rejection. It is very likely that they also contribute to acute allograft destruction. So far it has been impossible to distinguish the function of monocytes from that of macrophages, because cell phenotypes and their alterations upon activation are ill-defined. We have thus begun to characterize the ex vivo phenotype and function of rat monocytes in the normal state and during renal allograft rejection. Monocytes are recovered from both the central and the marginal blood pool by perfusing either the recipient's circulation or the allograft vasculature. Rat monocytes have a unique surface phenotype. During allograft rejection or after infusion of interferon-gamma they up-regulate class II MHC molecules, CD161 (NKR-P1A), CD62L and CD8, while CD4 and CD43 are down-modulated. Activated perfusate monocytes exert increased in vitro cytotoxicity against tumour targets, which differs from that of NK cells. We speculate that activated monocytes contribute to kidney allograft destruction by directly damaging endothelial cells or by promoting intravascular coagulation. PMID- 12086321 TI - Immunobiology of dendritic cells in the rat. AB - Dendritic cells (DC) are crucial orchestrators of the adaptive immune response. They are highly effective antigen samplers and have a unique ability amongst antigen presenting cells to activate naive T lymphocytes and subsequently direct the quality of the immune response. Understanding how DC initiate and regulate immune responses requires in-depth knowledge of DC function at cellular and molecular levels. Research on the biology of DC has predominantly used in vitro generated and ex vivo-isolated DC from mice and humans. It is, however, often difficult to relate such DC to those that actually exist in vivo. The model we have developed in the rat permits analysis of DC in a near-physiological setting, and provides a description of DC biology that other systems must take into account. In this review we focus on our own research on DC in the gastro intestinal tract, covering a variety of concepts in DC biology, and relate our findings to the work of others, to provide an overall picture of what is known regarding the nature of this complex cell type in the rat. PMID- 12086323 TI - Thymic development and repertoire selection: the rat perspective. AB - This review summarizes our current knowledge of T-cell maturation and repertoire selection in the rat thymus. Some unique features of early thymocyte development and of CD4/CD8 lineage decision are described. A detailed analysis of lineage progression through the CD4, CD8 "double positive" compartment and T-cell receptor-induced CD8 T-cell maturation in cell culture is provided. A second emphasis is placed on interactions between germline-encoded T-cell receptor elements with MHC molecules in thymic repertoire selection and alloreactivity PMID- 12086322 TI - Biochemical analysis of the lymphocyte cell surface--from alloantisera to the role of membrane proteins. AB - The rat has been invaluable for many aspects of the molecular analysis of the lymphocyte cell surface. One advantage of the rat over mice is the ability to obtain cells in large numbers for biochemical analysis together with the availability of many disease models. This review outlines some of the key findings in this laboratory over the last 25 years together with the technological advances that may be of relevance in this new post-genomic era where the goal is to understand the functions of the proteins coded by the multitude of genes now identified. PMID- 12086324 TI - The RT6 system of the rat: developmental, molecular and functional aspects. AB - RT6 is a developmentally regulated cell-surface membrane adenosine 5'-diphosphate ribosyltransferase/nicotinamide adenine dinucleotide-glycohydrolase inserted within the membrane by a glycophosphatidylinositol anchor. In the rat it is restricted to mature T lymphocytes and a subpopulation of natural killer cells. With respect to the data now available, three aspects concerning the function of RT6 are discussed: first, the meaning of the marked polymorphisms; second, its enzymatic activity; third, its possible role concerning T-cell survival. The observation that the rat RT6 gene contains two transcription start sites suggests their different use by distinct subpopulations of T cells. The fact that the expression of RT6 is defective in lymphopenic diabetes prone (DP-BB) rats, although the RT6 gene is structurally not grossly altered in these animals, makes this rat strain a promising model to study the biological meaning of RT6. While it mostly is believed that the RT6 expression defect of the DP-BB rat is a consequence of the lymphopenia, the present paper discusses the possibility that the RT6 expression defect is causally involved in the lymphopenia, and that a normal expression of RT6 may protect the recent thymic emigrants from apoptosis. PMID- 12086325 TI - Comparison of CT scanograms and cephalometric radiographs in craniofacial imaging. AB - OBJECTIVE: To compare measurements from human skulls and their images from cephalometric radiographs (CR) and computed tomography (CT) scanograms, in order to gauge the potential clinical use of the latter. DESIGN: Based on specific inclusion criteria, including stable centric occlusion, 13 adult skulls were selected from a larger collection. The mandible was taped to the maxilla after securing the occlusion of teeth and condylar seating in the glenoid fossa. Lateral and posteroanterior cephalographs and CT 'scout views' were taken of each skull by standardized methods. Landmarks were identified on skulls and images. OUTCOME MEASURES: Linear measurements were made on all three records; angular measurements only on CR and CT images. Intraclass correlation coefficients (r) were calculated to assess similarity among records. Paired t-tests were used to compare differences between mean measurements. RESULTS: No statistically significant differences were noted between mean angular values on CR and CT views (P > 0.05). The highest correlations were observed for several vertical midline distances between CT and direct skull measures: 0.82 < r < 0.995-greatest for nasion-menton. For sagittal distances, the highest correlation was between the direct measure of condylion-pogonion and its CR image (r= 0.73). Correlations between CR and skull transverse measures were higher (0.46 < r < 0.80) than the corresponding skull vs. CT measures (0.06 < r < 0.38). CT and CR images are 2D slices and projections, respectively, of 3D structures. Vertical CT and skull measures correspond because the CT projection reflects a 1:1 ratio in the midsagittal plane; CT projected lateral images are smaller than the skull measures. The CR image reflects a distortion (approximately 8%) that brings Co-Pg closer to its anatomic distance, inadvertently contributing to better clinical planning, particularly in orthognathic surgery. The pattern of distortion of PA images was in opposite directions for CR and CT views. CONCLUSIONS: Cephalograms and CT scanograms are close in depicting angular relations of structures, but they differ in the accuracy of imaging linear measurements, because the location and size of an object within the imaged 3D structure varies with both records. Logistic and economic considerations favor the use of cephalographs. PMID- 12086326 TI - Upper molar distalization: a critical analysis. AB - Traditional upper molar distalization techniques require patient co-operation with the headgear or elastics. Recently, several different intraoral procedures have been introduced to minimize the need for patient co-operation. This article reviews the appliances currently available for maxillary molar distalization and critically analyses their dentoalveolar and skeletal effects. PMID- 12086327 TI - Comparison of the expression patterns of two LIM-homeodomain genes, Lhx6 and L3/Lhx8, in the developing palate. AB - OBJECTIVES: To compare and contrast the gene expression of two LIM-homeobox type transcription factors, Lhx6 and L3/Lhx8, in secondary palate formation. METHODS: In situ hybridization histochemistry with digoxygenin (DIG) labelled cRNA probes specific for Lhx6 and L3/Lhx8. MATERIALS: Serial cryo-sections of embryonic day (E)13.5, 14.5, and 15.5 mice (C57BL/6). OUTCOME MEASURE: Comparison of the signal intensities of NBT/BCIP precipitate by alkaline phosphatase conjugated anti-DIG antibody. RESULTS: From E13.5 to E15.5, Lhx6 and L3/Lhx8 signals are detected in palatal mesenchyme, but the L3/Lhx8 signal is much more intense than the Lhx6 signal. In palatal epithelium, covering the mesenchyme, Lhx6 mRNA is transiently expressed at E14.5, while L3/Lhx8 mRNA expression is never detected throughout the development. CONCLUSION: Lhx6 and L3/Lhx8 functions may be partially redundant in the mesenchyme of the secondary palate, but not in the palatal epithelium. PMID- 12086328 TI - Analysis of facial growth in subjects with syndromic ectodermal dysplasia: a longitudinal analysis. AB - OBJECTIVE: To examine the craniofacial growth pattern of patients diagnosed with syndromic ectodermal dysplasia. DESIGN: Mixed longitudinal analysis of lateral cephalograms. SETTING: The data were analysed using a multilevel modelling technique with the MLwiN application software and the results presented numerically and graphically. SAMPLE POPULATION: All 61 subjects had severe hypodontia with the number of absent teeth (excluding third molars) ranging from 6 to 28 (mean = 15.4). At presentation the subjects had a mean age of 133 months and were followed longitudinally for between one and five subsequent occasions (mean 2.66 occasions; mean age at completion of observation 207 months). OUTCOME MEASURES: Lateral cephalograms taken at each visit. EXPERIMENTAL VARIABLES: Analysis of four angular measurements and four linear measurements, together with one calculated ratio. RESULTS: Growth curves are presented showing the trends of craniofacial growth. CONCLUSIONS: The most significant findings were for a universal tendency for the individuals to undergo a change in sagittal relationship of the jaws, becoming markedly more Class III with time. We have also demonstrated a significant difference in growth between the anterior and posterior face heights indicating that the subjects have a tendency to an anterior growth rotation. PMID- 12086329 TI - Spatiotemporal expression of NGFR during pre-natal human tooth development. AB - OBJECTIVES: The relation between nerve growth factor receptor (NGFR) in the human pre-natal tooth buds and the dental follicle was investigated. In particular, we sought to determine if there is a specific pattern of p75NGFR expression in developing human tooth buds and their surrounding tissue. SETTING AND SAMPLE POPULATION: The Department of Orthodontics at Copenhagen University, Denmark. Histological sections from 11 fetuses, aged 11-21 gestational weeks. METHOD: The sections were studied by conventional immunohistochemistry. RESULTS: Specific spatiotemporal patterns of p75NGFR reactions were observed in the tooth buds and dental follicle: Before matrix production by the ameloblasts, the entire inner enamel epithelium and the entire dental follicle display p75NGFR immunoreactivity; after matrix production is initiated, the immunoreactivity of the matrix producing cells is lost, as is that of the dental follicle adjacent to these matrix-producing cells. CONCLUSION: A unique spatiotemporal distribution of NGFR in the pre-eruptive human tooth bud was demonstrated. PMID- 12086330 TI - Cranial base changes following coronal suturectomy in craniosynostotic rabbits. AB - It has been suggested that surgical release of synostosed sutures may ameliorate various cranial base abnormalities in craniosynostotic patients. The present study was designed to test this hypothesis in a rabbit model with familial coronal suture synostosis (CSS). Data were collected from 56 New Zealand White rabbits: 32 unaffected controls, 11 with unoperated CSS, and 13 with CSS released by suturectomy performed at 25 days of age. Serial radiographs were taken at 25, 42 and 84 days. Linear, angular and triangular shape cranial base measurements were compared using ANOVA and tensor biometric analysis. Results revealed that at 84 days, both groups of CSS rabbits had significantly (p < 0.05) different anterior and total cranial base lengths, flatter cranial base angles, and dysmorphic anterior cranial base shapes when compared with normals. Significant (p < 0.05) differences were noted only for palatal and cranial base angles and posterior cranial base shape between CSS rabbits with and without suturectomy. However, significant (p < 0.05) changes were noted between pre- and postoperative measurements in posterior and total cranial base lengths and anterior and posterior cranial base shapes in CSS rabbits with suturectomy. Results revealed that surgical release of synostosed coronal sutures through suturectomy did not normalize cranial base growth patterns in CSS rabbits. These findings may be explained by the relatively late age of surgical release or suturectomy site resynostosis with continued dysmorphic cranial base growth. Alternatively, cranial base abnormalities seen in CSS rabbits may be early primary malformations, not secondary deformations amenable to surgical modification. PMID- 12086331 TI - Professor Moshe Wolman: pioneer in histochemistry. PMID- 12086332 TI - Renoprotective effects of valsartan and enalapril in STZ-induced diabetes in rats. AB - Effects of the angiotensin II type 1 (AT1) receptor antagonist valsartan and the angiotensin-converting enzyme (ACE) inhibitor enalapril were studied in streptozotocine (STZ)-induced diabetes in rats on the basis of microalbuminuria (Ma) and renal morphology. Five groups of Wistar rats were used, one group was the non-diabetic control, one group consisted of untreated STZ-diabetics and 3 groups of STZ-diabetics were treated with either enalapril and/or valsartan for 30 days. Blood glucose (BG) and Ma levels, body and kidney weight and glomerular size were measured. Immunohistochemical staining with an anti-transforming growth factor-beta1 (TGF-beta1) antibody was performed as well. In STZ-diabetics, BG and Ma levels were significantly increased when compared with the non-diabetic group. Although Ma levels in the valsartan-treated group was found to be higher than those in the non-diabetics group after 15 days of treatment, in all treated diabetic groups Ma levels were significantly decreased as compared with STZ diabetics at the end of the experiment. Thickening of the glomerular and tubular basement membranes, increased mesangial matrix and glomerular size were found in the untreated diabetic group. All these changes were less in the treated groups. A significant increase in TGF-beta1 immunoreactivity was found in glomeruli of untreated STZ-diabetics as compared with non-diabetics. Again, TGF-beta1 expression was decreased in the treated groups as compared with untreated STZ diabetics. We conclude that valsartan and enalapril have renoprotective effects in diabetic nephropathy. A combined therapy has an advantage because lower dosages of these drugs can be used. Their beneficial effects are related to a blockade of the renin-angiotensin system (RAS) and a decrease in TGF-beta1 expression in glomeruli. PMID- 12086333 TI - A histochemical study of the regeneration process after injury by pulsed Nd:YAG laser irradiation of root canals. AB - Histological changes of rat dental pulp cells were followed after injury. The regeneration process after 3, 6 and 10 days was monitored. Mandibular incisors were irradiated with a pulsed Nd:YAG laser at 2 W and 20 pulses per sec (pps) for 5 sec and the pulp was examined histologically and immunohistochemically for TGF beta1. Eruption of the developing tooth was disturbed for a short period only. Rapid formation of osteodentin was observed. After 3 days, a zone of fibrodentin matrix as well as newly formed vessels were found. Afterwards, regenerative dentin formation was observed accompanied by the formation of a layer of odontoblast-like cells in the damaged area. Immunohistochemical staining of TGF beta1 showed that positivity was present in small tissue areas beneath the mantle dentin, the zone of fibrodentin matrix and odontoblast-like cells. These results indicate that pulsed Nd:YAG laser irradiation of rat incisor pulp induces formation of osteodentin, and TGF-beta1 plays a role during regeneration. PMID- 12086334 TI - The expression of dystrophin, alpha-sarcoglycan, and beta-dystroglycan during skeletal muscle regeneration: immunohistochemical and western blot studies. AB - We evaluated re-expression of dystrophin, alpha-sarcoglycan and beta-dystroglycan in regenerating skeletal muscles of rats after cardiotoxin-induced myonecrosis in order to understand the dynamic behaviour of these proteins during the regeneration process. Immunohistochemical staining of these proteins almost disappeared in the sarcolemma of necrotic fibers on the 1st day, and was obscured due to non-specific staining on the 3rd day. Dystrophin was labeled faintly at the sarcolemma of regenerating muscle fibers on the 5th day. From the 5th day to the 10th day, levels of immunostaining of dystrophin increased. After the 14th day, dystrophin was stained conspicuously. alpha-Sarcoglycan was labeled weakly at the sarcolemma of small regenerating muscle fibers on the 5th day and was labeled conspicuously after the 7th day. beta-Dystroglycan was labeled moderately at the sarcolemma of regenerating muscle fibers on the 5th day and was labeled conspicuously after the 7th day. In western blot analysis, beta-dystroglycan persisted throughout the entire cycle of myonecrosis and regeneration, and re expression of alpha-sarcoglycan progressed faster than that of dystrophin. We speculate that regeneration advances from the basement membrane side to the subsarcolemmal side, and that proteins at the basement membrane side resist disruption and have a high capacity for regeneration. PMID- 12086335 TI - Alterations in tight junction molecules of uterine epithelial cells during early pregnancy in the rat. AB - Distribution patterns of the tight junction associated proteins ZO-1, claudin-1 and occludin were investigated in rat uterine epithelial cells during early pregnancy. Light microscopy and immunohistochemical labelling were used to detect these proteins on days 1, 3, 6 and 7 of pregnancy. Intense staining of claudin-1 at the apical region of the lateral plasma membrane accompanied diffuse staining throughout the cytoplasm. ZO-1 was also localised in the apical region, but ZO-1 was not present in the lower two thirds of the lateral plasma membrane or in the cytoplasm. Occludin was present only on days 6 and 7 of pregnancy. Labelling was also localised in the apical region of the lateral plasma membrane where tight junctions are known to be present. Our results show that ZO-1, claudin-1 and occludin are present in the apical region of uterine epithelial cells, and appear to play a role in the very dynamic tight-junctional network of uterine epithelial cells during early pregnancy. In particular, occludin appears only during uterine receptivity for implantation. PMID- 12086336 TI - Expression of intermediate filaments and desmosomal proteins during differentiation of the human spinal cord. AB - Differentiation of the human spinal cord and involution of its caudal end were investigated in 4-9-week human conceptuses using immunofluorescence and electron microscopy. In the spinal cord, several types of intermediate filament proteins and desmoglein were expressed in parallel: in early stages (4 to 6 weeks), neurofilaments were expressed in low amounts only in the neuroblast processes of the marginal layer. At 6 weeks, differences in staining intensity and distribution patterns of neurofilaments became apparent between lumbar and sacrococcygeal (tail) parts of the spinal cord. Neurofilament expression increased in the mantle and marginal layers of the lumbar spinal cord coinciding with advancing neurogenesis. In contrast, neurofilament expression decreased in the sacrococcygeal spinal cord in association with regression of all tail organs. Regression was characterized by the appearance of large amounts of dead cells and macrophages. Strong vimentin expression was found in neuroepithelial (ependymal) cells and in the radial glia of the spinal cord throughout all stages examined. Coexpression of vimentin and glial fibrillary acidic protein was found only in the radial glia in the earliest developmental stage. Desmoglein was expressed in low amounts around the central canal which was probably associated with the immature junctional complexes that were present between ependymal cells. In conclusion, temporal and spatial distribution patterns of intermediate filament proteins in specific cell populations characterizes differentiation and caudal involution of the human spinal cord. PMID- 12086337 TI - Cartilage-specific matrix protein, chondromodulin-I (ChM-I), is a strong angio inhibitor in endochondral ossification of human neonatal vertebral tissues in vivo: relationship with angiogenic factors in the cartilage. AB - Although cartilage contains many angiogenic factors during endochondral ossification, it is an avascular tissue. The cartilage-specific non-collagenous matrix protein chondromodulin-I (ChM-I) has been shown to be a strong angio inhibitor. To elucidate whether ChM-I plays an essential role in angio-inhibition during endochondral ossification in man, we investigated the expression and localization of ChM-I in comparison with those of angiogenic factors and the endothelial cell marker CD34 in human neonatal vertebral tissues. Although invasion of CD34-positive endothelial cells was observed in primary subchondral spongiosa, expression of the marker of endothelial cells, CD34, was not found in neonatal vertebral cartilage matrix. Type II collagen was deposited in all matrices during endochondral ossification, whereas aggrecan was deposited in the matrix of hypertrophic cartilage, especially around lacunae. Vascular endothelial growth factor (VEGF), which is known to be a strong angiogenic factor, was localized in chondrocytes in mature to hypertrophic cartilage and also in bone marrow. Fibroblast growth factor-2 (FGF-2; basic fibroblast growth factor), which is also known to be a strong angiogenic factor, was localized in the cytoplasm of chondrocytes of mature cartilage in human vertebral cartilage tissues. Transforming growth factor (TGF)-beta has been reported to have many functions including angiogenesis, and TGF-beta1 was also localized in mature chondrocytes in endochondral tissues undergoing ossification. On the other hand, the novel cartilage-specific matrix protein ChM-I was localized in interterritorial regions of the matrix in mature to hypertrophic cartilage, especially around lacunae. In conclusion, these observations indicate that ChM-I may serve as a barrier against the angiogenic properties of VEGF, FGF-2 and TGF-beta1 during endochondral ossification, and this matrix molecule may play an essential role in determining the avascular nature of cartilage in vivo. PMID- 12086338 TI - Expression of ICAM-1, VCAM-1, E-selectin and TNF-alpha on the endothelium of femoral and iliac arteries in thromboangiitis obliterans. AB - Immunohistochemical light and electron microscopical analysis of surgical biopsies obtained from femoral and iliac arteries of patients with thromboangiitis obliterans (TAO) were performed to investigate the presence of tumour necrosis factor-alpha (TNF-alpha) and expression of the endothelial cell adhesion molecules intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-selectin. Expression of ICAM-1, VCAM-1 and E selectin was increased on endothelium and some inflammatory cells in the thickened intima in all TAO patients. Ultrastructural immunohistochemistry revealed contacts between mononuclear blood cells and ICAM-1-, and E-selectin positive endothelial cells. These endothelial cells showed morphological signs of activation. The present data indicate that endothelial cells are activated in TAO and that vascular lesions are associated with TNF-alpha secretion by tissue infiltrating inflammatory cells, ICAM-1-, VCAM-1- and E-selectin expression on endothelial cells and leukocyte adhesion via their ligands. The preferential expression of inducible adhesion molecules in microvessels and mononuclear inflammatory cells suggests that angiogenesis contributes to the persistence of the inflammatory process in TAO. PMID- 12086339 TI - Mast cells and inflammatory mediators in chronic ulcerative colitis. AB - Chronic ulcerative colitis (CUC) is an inflammatory destructive disease of the large intestine characterized by motility and secretion disorders. In the past decade, attention has been paid to the role of neuronal structures and mast cells in regulating inflammatory and immune responses in inflammatory bowel disease (IBD). The present study was performed to demonstrate neuronal fibres (NF) and cells containing substance P (SP), tryptase and serotonin (SER) in the colonic wall of patients with CUC in remission. Biopsy specimens of 6 patients with CUC were investigated with immunocytochemical methods. Normal colon tissue obtained from 6 patients with rectal carcinoma was used as a control. An increased number of SP- and SER-positive NF was found in all the layers of the intestinal wall. The number of SER-containing endocrine cells in the mucosal glands was also increased per crypt. Tryptase-, SP- and SER-immunopositive mast cells were found in higher amounts than in control specimens in close apposition to the basal lamina of the glands among the epithelial cells and in other layers of the gut wall. Two types of mast cells were found: mast cells containing both tryptase and SP, and mast cells containing tryptase only. It is concluded that interactions between neuronal elements and mast cells play a significant role in the progress and maintenance of inflammatory processes in CUC. PMID- 12086340 TI - Microdensitometric assay of enzymatic activities in parthenogenetically activated and in vitro fertilized bovine oocytes. AB - To examine the paternal genome's role in reprogramming metabolic activity in one cell embryos, we investigated metabolic aspects of bovine oocytes after in vitro maturation and in vitro fertilization and after in vitro parthenogenetic activation with a Ca2+ ionophore and 6-dimethylaminopurine. We assayed succinate dehydrogenase, lactate dehydrogenase and glucose-6-phosphate dehydrogenase activities by microspectrophotometry in immature oocytes and oocytes after maturation, in vitro fertilization and parthenogenetic activation. Succinate dehydrogenase activity significantly increased after in vitro maturation, significantly decreased after Ca2+ ionophore activation and further decreased after 6-dimethylaminopurine treatment. Lactate dehydrogenase activity showed a significant decrease in bovine oocytes after in vitro maturation, remained unchanged in Ca2+ ionophore-treated oocytes and rose significantly after 6 dimethylaminopurine treatment. This activity was dramatically reduced after in vitro fertilization, reaching absorbance levels that were not different from those in mature and Ca2+ ionophore-treated oocytes. Glucose-6-phosphate dehydrogenase activity was significantly lower in matured oocytes as compared to immature oocytes, was significantly higher after artificial activation with Ca2+ ionophore and remained constant after 6-dimethylaminopurine treatment or after in vitro fertilization. We suggest that metabolic changes involved in parthenogenetic activation are similar to those occurring after fertilization. PMID- 12086341 TI - Histochemical and ultrastructural study of the chicken salivary palatine glands. AB - Salivary glands are a good model to investigate the relationship between cell secretion and glandular structure. Most studies of this organ deal with mammals, but we are interested in a morphofunctional characterization of these glands in poultry in relation with particular feeding habits. For this purpose, conventional and lectin histochemical methods as well as ultrastructural methods have been applied to the chicken lateral and medial palatine salivary glands. It was found that periodic acid-Schiff (PAS)-positive, alcianophilic, and metachromatic or orthochromatic cells were present with a more homogeneous distribution pattern in lateral glands than in medial palatine glands. Lectin staining depended on the lectin type that was applied, but also on the glandular part both in lateral and medial glands. Ultrastructural studies showed cytoplasmic membranous structures with a scattered granular or filamentous content depending on the secretory cell. In conclusion, morphofunctional characteristics of salivary glands of chicken suggest that their products are involved in lubrication and humidification of food ingested, and probably in protection of the oral surface, as has been previously described for other animals showing similar histochemical staining patterns. PMID- 12086343 TI - The impact of bone turnover and bone-active agents on bone quality: focus on the hip. PMID- 12086342 TI - Glucidic determinants expressed by the digestive apparatus of Umbrina cirrosa (L.) fries as revealed by lectin histochemistry. AB - Glycoconjugates present in the various parts of the digestive apparatus (oesophagus, stomach, intestine) of 27, 34 and 44 days-old Umbrina cirrosa (L.) fries were characterised by means of lectin histochemistry in conjunction with sialidase digestion and KOH treatment. High amounts of carbohydrates were detected especially in goblet cells of the oesophagus. Positivity obtained with the various lectins in oesophagus, stomach and intestine did not change during the development of the fries examined. D-Gal and D-GalNAc appeared to occupy a terminal position in oligosaccharidic chains whereas they are linked to sialic acid residues in adult subjects, which suggests an incomplete chemical structure of the glycoconjugates secreted by the digestive apparatus of fries. PMID- 12086344 TI - Skeletal status in children, adolescents and young adults with end-stage renal failure treated with hemo- or peritoneal dialysis. AB - The skeletal status in 30 children, adolescents and young adults (18 females, 12 males) with end-stage renal failure (ESRF) aged 9-23 years (mean 15.8 +/- 3.6 years) was evaluated using measurements of bone mineral density (BMD, g/cm2) at the spine and total body (TB) (Lunar DPX-L, USA), quantitative ultrasound (QUS) of the hand phalanges (DBM Sonic 1200, IGEA, Italy) and laboratory investigations (parathyroid hormone, serum total and ionized calcium, serum phosphate). Eleven subjects were treated with hemodialysis and 19 with peritoneal dialysis. The mean value of the amplitude-dependent speed of sound (Ad-SoS, m/s) measured by QUS was significantly decreased in comparison with the value obtained in a group of 686 age-matched controls (1942 +/- 74 m/s vs 2050 +/- 77 m/s, p<0.0001). BMD measurements were also decreased in comparison with mean values for the healthy population (Z-scores for spine -1.47, and for TB -1.53). Duration of dialysis correlated significantly with spine-BMD, TB-BMD and Ad-SoS (r = -0.37, r = -0.45, r = -0.55, respectively, p<0.05), while duration of ESRF did not have such an influence. Laboratory investigations did not correlate with skeletal parameters. Ad-SoS correlated significantly with spine-BMD (r = 0.45, p<0.05) and TB-BMD (r = 0.56, p<0.01). Both QUS and BMD values correlated significantly with Tanner stages (r ranged from 0.59 to 0.69, p<0.001) and did not increase with age except for correlation between age and TB-BMD. In conclusion, skeletal status in the population studied is strongly affected by ESRF. Both QUS and BMD measurements show an ability to express skeletal changes in a similar manner, though the QUS parameter seems to be more sensitive at revealing changes due to renal failure. PMID- 12086345 TI - Digital X-ray radiogrammetry predicts hip, wrist and vertebral fracture risk in elderly women: a prospective analysis from the study of osteoporotic fractures. AB - Digital X-ray radiogrammetry (DXR) is a technique that uses automated image analysis of standard hand radiographs to estimate bone mineral density (DXR-BMD). Previous studies have shown that DXR-BMD measurements have high precision, are strongly correlated with forearm BMD and are lower in individuals with prevalent fractures. To determine whether DXR-BMD measurements predict wrist, hip and vertebral fracture risk we conducted a case-cohort study within a prospective study of 9704 community-dwelling elderly women (the Study of Osteoporotic Fractures). We compared DXR-BMD, and BMD of the radius (proximal and distal), calcaneus, femoral neck and posteroanterior lumbar spine in women who subsequently suffered a wrist (n = 192), hip (n = 195), or vertebral fracture (n = 193) with randomly selected controls from the same cohort (n = 392-398). DXR BMD was estimated from hand radiographs acquired at the baseline visit. The radiographs were digitized and the Pronosco X-posure System was used to compute DXR-BMD from the second through fourth metacarpals. Wrist fractures were confirmed by radiographic reports and hip fractures were confirmed by radiographs. Vertebral fractures were defined using morphometric analysis of lateral spine radiographs acquired at baseline and an average of 3.7 years later. Age-adjusted odds ratio (OR, vertebral fracture) or relative hazard (RH, wrist and hip fracture) for a 1 SD decrease in BMD were computed. All BMD measurements were similar for prediction of wrist (RH = 1.5-2.1) and vertebral fracture (OR = 1.8-2.5). Femoral neck BMD best predicted hip fracture (RH = 3.0), while the relative hazards for all other BMD measurements were similar (RH = 1.5-1.9). These prospective data indicate that DXR-BMD performs as well as other peripheral BMD measurements for prediction of wrist, hip and vertebral fractures. Therefore, DXR-BMD may be useful for prediction of fracture risk in clinical settings where hip BMD is not available. PMID- 12086346 TI - Fractal analysis of trabecular bone texture on calcaneus radiographs: effects of age, time since menopause and hormone replacement therapy. AB - An analysis of trabecular bone texture based on fractal mathematics, when applied to trabecular bone images on plain radiographs, can be considered as a reflection of trabecular bone microarchitecture. It has been shown to be able to distinguish postmenopausal osteoporosis cases from controls. This cross-sectional study was carried out to investigate the influence of age, time since menopause and hormone replacement therapy (HRT) on the fractal dimension of trabecular bone texture at the calcaneus in a sample of 537 healthy women. Fractal analysis of texture was performed on calcaneus radiographs and the result expressed as the Hmean parameter (H = 2-fractal dimension). Total hip, femoral neck and lumbar spine bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry. There was a statistically significant Hmean parameter decrease with age (p<0.0001) but the degree of correlation was low (r = -0.2) compared with the correlation between age and BMD (r = -0.36 to -0.61 according to the BMD site). We found a weak but statistically significant correlation between time since menopause and Hmean (r = -0.14, p = 0.03) in the 241 postmenopausal women included in the study. Hmean was significantly lower in a group of postmenopausal women without HRT (n = 110) compared with a group of age-matched postmenopausal women with HRT (n = 110): respectively 0.683 +/- 0.043 and 0.695 +/- 0.038 (p = 0.03). In conclusion, this study suggests that there is a menopause- and age related decrease in the Hmean parameter and that HRT interferes with the results of the fractal analysis of trabecular bone texture on calcaneus radiographs. PMID- 12086347 TI - Mandibular radiomorphometric indices in the diagnosis of reduced skeletal bone mineral density. AB - Diagnosis of osteoporosis allows the delivery of preventive and therapeutic intervention and is usually achieved using bone densitometric techniques. One referral criterion for densitometry is osteopenia on radiographs. The aim of this study was to measure the validity of mandibular cortical indices measured on panoramic radiographs in the diagnosis of reduced skeletal bone density. Seventy four women underwent bone densitometry of the femoral neck, lumbar spine and the forearm. Fifty-five patients (74%) were classified as having a reduced bone density (T-score < or = -1). Twenty-seven patients had a T-score of < -2.5 observed at one or more of the three measurement sites. A panoramic radiograph was taken of each patient and two observers made measurements of cortical thickness at the mental foramen (mental index, MI), antegonion (antegonial index, AI) and gonion (gonial index, GI) regions. Logistic regression and receiver operating characteristic (ROC) curve analyses were used to measure the validity of cortical indices in the diagnosis of reduced bone mineral density. Only MI contributed significantly to a diagnosis of low skeletal bone mineral density (T score < or = -1). The 95% limits of agreement between observers in measurement of MI were 1.32 to +1.32 mm. When data for both observers were combined, the area under the ROC curve was 0.733 (SE = 0.072; 95% confidence interval = 0.618 to 0.83), indicating moderate accuracy. A diagnostic threshold for MI of 3 mm (or less) is suggested as the most appropriate threshold for referral for bone densitometry. However, the study provided only limited support for the use of panoramic radiomorphometric indices in diagnosing low skeletal bone mineral density. They might, questionably, be used as part of a method of osteoporosis risk assessment. PMID- 12086348 TI - Bone mass, bone metabolism, gonadal status and body mass index. AB - Weight and gonadal status are the main determinants of bone mass in women. Because of this it is important to study which influences it more. The effect of weight (expressed as body mass index, BMI) and gonadal status of women on total body bone mineral content (TBBMC) and regional bone mineral content (BMC) was investigated. A total of 373 normal women (mean age 48.9 +/- 13.4 years) were studied: 171 postmenopausal women (mean age 59.3 +/- 9.5 years; years since menopause 11.3 +/- 6.7 years); 76 perimenopausal women (mean age 48.9 +/- 2.2 years); and 126 premenopausal women (mean age 34.7 +/- 7.4 years). In all the women, TBBMC and regional BMC were determined by dual-energy X-ray absorptiometry. Also biochemical markers of bone metabolism (total alkaline phosphatase and tartrate-resistant acid phosphatase) and serum estrone and estradiol were determined. When the women were stratified by gonadal status and BMI, thin women (BMI <20 kg/m2) had significantly lower TBBMC and regional BMC, lower gonadal steroid concentration and higher levels of biochemical markers than overweight (BMI 25-30 kg/m2) and obese (BMI >30 kg/m2) women, regardless of gonadal status. Overweight and obese women had findings suggestive of increased parathyroid activity, but greater bone mass. Weight rather than gonadal steroid concentration is the main determinant of bone mass in women regardless of gonadal status. PMID- 12086349 TI - Quantitative ultrasound of bone in male osteoporosis. AB - Quantitative ultrasound (QUS) measurement, a different approach to bone fragility assessment, has already been attempted in women with osteoporosis but rarely in men. In order to test its value and ability to identify osteoporotic men, a case control prospective study was conducted using the Lunar Achilles, a device that measures attenuation and velocity parameters. Broadband ultrasound attenuation (BUA), speed of sound (SOS) and stiffness index (SI), a composite parameter, were assessed through the heel of 66 osteoporotic patients, and compared with the results in 35 controls. Patients had sustained a low-trauma fracture and/or had a lumbar and/or femoral bone mineral density (BMD) more than 2.5 SD below the young male reference value. As expected, all QUS parameters were statistically lower in patients, as were the dual-energy X-ray absorptiometry (DXA) measurements at the hip and lumbar spine. The two methods were compared for their ability to predict the risk of osteoporotic fractures. The odds ratios (ORs), with their 95% confidence limits, for fractures per 1 SD decrease were significant, especially for SOS and SI (OR = 2.3 [1.4-3.6] and 2.1 [1.3-3.3] respectively) and to a lesser extent for BUA (1.6 [1.0-2.4]). Our study suggests that QUS is associated with a history of low-trauma fracture in men; sensitivity is, however, less than when results are compared with BMD measurements (OR = 2.8 [1.6-5.0] and 3.4 [1.6 7.0] for lumbar spine and hip, respectively). Prospective studies are required before QUS can be recommended for clinical use in male osteoporosis. PMID- 12086350 TI - Correlates of osteoprotegerin levels in women and men. AB - Osteoprotegerin (OPG) is a potent antiresorptive molecule that binds the final effector for osteoclastogenesis, receptor activator of NF-kappaB ligand (RANK-L). OPG production is regulated by a number of cytokines and hormones, including sex steroids, but there are few data on age and gender effects on circulating serum OPG levels, as well as possible relationships between OPG levels and bone turnover markers or bone mineral density (BMD). Thus, we measured serum OPG levels in an age-stratified, random sample of men (n = 346 age range, 23-90 years) and women (n = 304; age range 21-93 years) and related them to sex steroid levels, bone turnover markers and BMD. Serum OPG levels increased with age in both men (R = 0.39, p < 0.001) and women (R = 0.18, p < 0.01). Premenopausal women had higher OPG levels than men under age 50 years (171 +/- 6 pg/ml vs 134 +/- 6 pg/ml, respectively, p < 0.001), whereas serum OPG levels were no different in postmenopausal women compared with men = 50 years (195 +/- 7 pg/ml vs 188 +/- 7 pg/ml, respectively, p = 0.179). OPG levels correlated inversely with serum bioavailable testosterone levels in men = 50 years (R = -0.27, p < 0.001), but no associations were present with either estrogen or testosterone levels in the women. In the men, there was a trend for OPG levels to be associated positively with bone resorption markers and inversely with BMD. Collectively, the gender difference in OPG levels suggests that sex steroids may regulate OPG production in vivo, as has been found in vitro. Moreover, OPG production may also rise with increases in bone turnover, probably as a homeostatic mechanism to limit bone loss. Further studies directly testing these hypotheses should provide additional insights into the potential role of OPG in bone loss related to aging and sex steroid deficiency. PMID- 12086351 TI - A clinical prediction rule to identify premenopausal women with low bone mass. AB - Identifying premenopausal women at risk for osteoporosis and related fractures is a potentially important way to reduce the burden of illness from this disease as low peak bone mass is a risk factor for postmenopausal osteoporosis. We examined predictors of 'low' peak bone mass in 668 healthy, pre-menopausal, Caucasian women ages 18-35 years. Predictors of bone mass were assessed using a detailed, standardized interview. Bone mass was assessed using two measures: dual-energy X ray absorptiometry (DXA) at the femoral neck and lumbar spine, and quantitative ultrasound (QUS) of the heel, which evaluates stiffness, speed of sound (SOS) and broadband ultrasound attenuation (BUA). Bone mass was considered 'low' if the corresponding Z-score was <-1.00 (DXA values, stiffness) or if values were in the lowest quintile (BUA, SOS). Using multivariate logistic regression modeling, predictors of low bone mass based on QUS, DXA or both were determined. The mean age of the cohort was 27.3 years. Independent predictors of low bone mass by both DXA and QUS were: low body weight, menarche at age 15 years or later and physical inactivity as an adolescent. Individuals with all three risk factors had a 92% chance of having low bone mass using both techniques. This suggests that a simple risk factor assessment can identify most young women with low peak bone mass. Early intervention in this group of women may reduce the risk for osteoporosis in later life. PMID- 12086352 TI - Bone changes due to glucocorticoid application in an ovariectomized animal model for fracture treatment in osteoporosis. AB - In a pilot experiment comparing four different modalities for inducing osteoporosis in the sheep, a combination of ovariectomy, calcium/vitamin D restricted diet and steroid administration was found to generate the highest decrease in bone mineral density (BMD). The aim of the present study was to quantify the outcome of this triple treatment in an animal model of osteoporosis in terms of alteration in bone mass, bone structure and bone mechanics. A total of 32 sheep were divided into two equal groups. Group 1 (age 3-5 years) was used as a normal control. Group 2 (age 7-9 years) was ovariectomized, fed a calcium/vitamin D-restricted diet and injected with methylprednisolone (MP) over 7 months (22 weeks MP solution, 6 weeks MP suspension). The BMD at the distal radius and tibia was determined preoperatively and at repeated intervals bilaterally using quantitative computed tomography. Steroid blood levels were determined 4 and 24 h after selected injections. BMD was measured at L3 and L4 after 7 months. Biopsies were taken from iliac crests, vertebral bodies and femoral heads, and bone structure parameters investigated by three-dimensional micro-CT. Compressive mechanical properties of cancellous bone were determined from biopsies of vertebral bodies and femoral heads. After 7 months of osteoporosis induction the BMD of cancellous bone decreased 36 +/- 3% in the radius and 39 +/- 4% in the tibia. Steroid blood levels 24 h after injection of MP suspension were significantly higher than after injection of MP solution. Changes in structural parameters of cancellous bone from the iliac crest, lumbar spine and femoral head in group 2 indicated osteoporosis-associated changes. In group 2 there was a significant reduction in BMD of the lumbar spine and a significant reduction in stiffness and failure load in compression testing of biopsies of lumbar vertebrae. In sheep, changes in the structural parameters of bone such as trabecular number and separation during osteoporosis induction are comparable to the human situation. The sheep model presented seems to meet the criteria for an osteoporosis model for fracture treatment with respect to mechanical and morphometric bone properties. PMID- 12086353 TI - Heel ultrasonography in monitoring alendronate therapy: a four-year longitudinal study. AB - The possibility of using quantitative ultrasound (QUS) in monitoring the response to antiresorptive drugs has yet to be defined. The aim of the present study was to evaluate whether heel ultrasonography, considering its characteristics of long term precision, is able to monitor osteoporotic patients treated with alendronate. We studied 150 postmenopausal osteoporotic women (age 59.6 +/- 5.3 years) treated with alendronate and calcium (n = 74) or with calcium alone (n = 76) for 4 years. At baseline and after 12, 24, 36 and 48 months, we measured bone mineral density (BMD) at the lumbar spine by dual-energy X-ray absorptiometry (DXA, Hologic 4500), and speed of sound (SOS), broadband ultrasound attenuation (BUA) and Stiffness at the calcaneus by Achilles plus. Moreover, the longitudinal precision of QUS parameters was assessed by measuring 10 subjects once a month for 1 year and, on the basis of the coefficients of variation we obtained, we calculated the Least Significant Change between two measurements. In the alendronate-treated patients, at year 1, BMD increased by 4.2%, SOS by 0.4%, BUA by 1.1% and Stiffness by 3.2%; at year 2, BMD increased by 5.0%, SOS by 0.7%, BUA by 1.4% and Stiffness by 5.7%. At year 3, BMD increased by 6.2%, SOS by 0.9%, BUA by 1.8% and Stiffness by 7.6%. At the end of the study period, BMD increased by 7.6%, SOS by 1.2%, BUA by 1.9% and Stiffness by 9.0%. The minimal significant difference between two measurements was 0.8% for SOS, 5.6% for BUA and 5.0% for Stiffness. Among the QUS parameters, Stiffness showed the greatest total treatment effect and a longitudinal sensitivity which was only slightly lower than BMD. The MTI, which represents the period between scans required to show that a 'true' change has occurred, was 1.8, 2.7, 11.9 and 2.2 years for BMD, SOS, BUA and Stiffness respectively. Therefore, although the spinal BMD remains the optimal method, QUS at the heel, and in particular Stiffness, seems to be a sensitive tool for monitoring the response to alendronate. PMID- 12086354 TI - Comparison of histomorphometric descriptors of bone architecture with dual-energy X-ray absorptiometry for assessing bone loss in the orchidectomized rat. AB - We have compared different methods for measuring bone loss in the orchidectomized (ORX) rat model of male osteoporosis: densitometry (DXA), ash weight, anatomic bone indices, histomorphometry and two-dimensional trabecular architecture analysis. Forty-eight male Wistar rats were studied at 2, 4, 8 and 16 weeks (four groups). In each group, 6 rats were ORX and 6 sham-operated were used as control. DXA was performed on the whole body, tibia and femur. Histomorphometry was performed on the secondary spongiosa of the tibia: trabecular bone volume (BV/TV) and trabecular characteristics (number, separation and thickness) were measured. Architecture analysis comprised strut identification, star volume of the marrow spaces and trabeculae, Euler-Poincare number (E) and Kolmogorov fractal dimension (Dk). Bone mineral densities of the whole body, tibia and femur were reduced at 16 weeks in the ORX group. BV/TV was significantly decreased in the ORX group from the fourth week. Differences in the sensitivity of the architectural methods were found. There were no differences in trabecular thickness nor in trabecular star volume between ORX and controls even after 16 weeks. E became different at 8 weeks. Trabecular number, node count, star volume of the marrow spaces and trabecular separation became significantly different at 4 weeks Dk was modified after 2 weeks (p < 0.05 at 2 weeks, p < 0.001 from 4 weeks). In the ORX model, Dk appeared the most potent descriptor of trabecular bone disorganization by revealing the earliest changes at the network level. PMID- 12086355 TI - Bone mineral density and metabolism in familial dysautonomia. AB - Familial dysautonomia (FD) patients suffer from multiple fractures and have reduced bone pain, which defers the diagnosis. The pathogenesis of bone fragility in FD is unknown. This study aimed to characterize bone mineral metabolism and density in FD. Seventy-nine FD patients aged 8 months to 48 years (mean age 13.9 +/- 10.4 years, median 12.3) were studied. Clinical data included weight, height, bone age, weekly physical activity and history of fractures. Bone mineral density (BMD) of the lumbar spine (n = 43), femoral neck (n = 26), total hip (n = 22) and whole body (n = 15) were determined by dual-energy X-ray absorptiometry. Serum 25 hydroxyvitamin D3, osteocalcin, bone alkaline phosphatase (B-ALP), parathyroid hormone and urinary N-telopeptide cross-linked type 1 collagen (NTx) were determined in 68 patients and age- and sex-matched controls. Forty-two of 79 patients (53%) sustained 75 fractures. Twenty-four of 43 patients had a spine Z score < -2.0, and 13 of 26 had a femoral neck Z-score < -2.0. Mean femoral neck BMD Z-score was lower in patients with fractures compared with those without ( 2.5 +/- 0.9 vs -1.5 +/- 1.0, p = 0.01). Mean body mass index (BMI) was 16 kg/m2 in prepubertal patients and 18.4 kg/m2 in postpubertal patients. Bone age was significantly lower than chronological age (75.5 vs 99.3 months in prepubertal patients, p < 0.001; 151 vs 174 in postpubertal patients, p < 0.05). NTx and osteocalcin levels were higher in FD patients compared with controls (400 +/- 338 vs 303 +/- 308, BCE/mM creatinine p < 0.02; 90 +/- 59.5 vs 61.8 +/- 36.9 ng/ml, p < 0.001, respectively). B-ALP was lower in FD patients compared with controls (44.66 +/- 21.8 vs 55.36 +/- 36.6 ng/ml, p < 0.04). Mean spine Z-score was significantly lower in physically inactive compared with active patients (-3.00 +/- 1.70 vs -1.77 +/- 1.3, respectively, p = 0.05). We conclude that fractures in FD patients are associated with reduced BMD. FD patients have increased NTx and osteocalcin. Contributing factors include reduced BMI, failure to thrive and reduced physical activity. Preventive therapy and early diagnosis are essential. PMID- 12086356 TI - Interest of a prescreening questionnaire to reduce the cost of bone densitometry. AB - Bone mineral density (BMD) measurement is widely recognized as the best single tool to identify patients with a high lifetime risk of developing an osteoporosis related fracture. However, the cost/benefit value of screening the whole population has been repeatedly challenged and demonstrated to be rather poor. In many countries, BMD scan is not or no longer reimbursed because of lack of validated criteria to identify patients who should benefit from this procedure. Based on the proposals of a nationwide expert panel, a simple questionnaire identifying historical, clinical and behavioral risk factors for osteoporosis was developed. The aim of this study was to assess the diagnostic accuracy of the proposed criteria; to determine the extent to which this questionnaire could be useful for optimizing the use of densitometry tests; and, more specifically, to estimate the diagnostic costs per osteoporotic or osteopenic patient detected. For this purpose, we applied the questionnaire to 3998 consecutive individuals at least 20 years old, of both genders, either consulting spontaneously or referred for a BMD measurement to an outpatient osteoporosis center. BMD was measured by dual-energy X-ray absorptiometry (DXA) at the lumbar spine and at the hip (both total hip and femoral neck). Diagnostic accuracies were evaluated through measures of sensitivity, specificity, and positive and negative predictive values. After determining a benchmark value for age, different strategies were compared in order to identify the most cost-effective one in terms of cost per patient detected. According to the WHO operational definition of osteoporosis (T score <-2.5), 31% of the subjects were classified as osteoporotic at one or more of the measured sites. If only patients with at least one of the proposed risk factors had been referred for scans, 33.3% of the BMD measurements would have been avoided. Among those, less than 5% were missclassified as they did have osteoporosis at the total hip and up to 23% at one or more of the considered sites. On the other hand, of the subjects who would be recommended for a densitometry test, only a small fraction were identified correctly (the positive predictive values varied from 11.3% at the total hip to 34.8% at any site). In this first setting, the suggested criteria seem useful chiefly for excluding subjects who do not need a DXA scan rather than selecting osteoporotic patients. When applied only to patients aged 61 years or more, the positive predictive values rose to 15.1% (total hip) and 42.9% (any site), whereas the corresponding negative predictive values were set at 93% and 68.6%. In comparison, with a mass screening scenario the estimated diagnostic costs (costs associated with the DXA procedure) per osteoporotic patient detected at any of the considered sites would be reduced by more than 9% (59.4 instead of 65.3 Euros) if the suggested indications are taken into account for prescreening patients. And when the questionnaire is applied only to women over the age of 60 years these costs would be further reduced to 50.6 Euros, representing a 23% decrease. Then, a prescreening strategy based on these indications concomitantly with an age selective criterion could represent a promising way toward a more rational use of BMD measurement. PMID- 12086357 TI - The association between allergic rhinitis and psychologic disturbances. PMID- 12086358 TI - Addressing occupational allergy to natural rubber latex among health care workers. PMID- 12086359 TI - Mammalian toll-like receptors. AB - OBJECTIVE: To review the role of mammalian Toll-like receptors (TLRs) in host defense. DATE SOURCES: MEDLINE search and current literature. RESULTS: First, TLRs participate in the recognition of molecular patterns present on microorganisms. Second, TLRs are expressed at the interface with the environment, the site of microbial invasion. Third, activation of TLRs induces expression of costimulatory molecules and the release of cytokines that instruct the adaptive immune response. Fourth, activation of TLRs leads to direct antimicrobial effector pathways that can result in elimination of the foreign invader. CONCLUSIONS: Maintained throughout evolution, mammalian TLRs are proteins that participate in innate immunity to microbial pathogens. Insights into TLR activation pathways provide new therapeutic targets for intervention in infectious and immunologic disease. PMID- 12086360 TI - Haemophilus influenzae type b meningitis in a fully immunized 2-year-old. PMID- 12086361 TI - Self-reported hay fever and panic attacks in the community. AB - OBJECTIVE: The objective of this study was to determine the relationship between self-reported hay fever and common mental disorders among adults in the general population. METHODS: Data were drawn from the Midlife Development in the United States Survey, a representative household survey of the adult US population (25 to 74 years old; n = 3,032). Multivariate logistic regression analyses were used to determine the relationship between self-reported hay fever and current major depression, panic attacks, generalized anxiety disorder, and alcohol/substance use disorders. RESULTS: Self-reported hay fever was associated with a significantly increased odds of panic attack (odds ratio = 1.8 [1.2, 2.6]), which persisted after adjusting for differences in sociodemographic characteristics and comorbid mental disorders. Self-reported hay fever was not associated with a significantly increased likelihood of major depression, generalized anxiety disorder, or alcohol/substance use disorders. CONCLUSIONS: Consistent with previous findings, these data show a relationship between self-reported hay fever and increased likelihood of panic attacks among adults in the general population. The mechanism of the observed association remains unknown. Future work that examines the relationship between hay fever and panic attacks, as well as other mental disorders using both self-report and objective measurement of allergic response in prospective, longitudinal, epidemiologic data may be useful in improving our understanding of this observed link. PMID- 12086362 TI - A hospital-based screening program for natural rubber latex allergy. AB - BACKGROUND: Natural rubber latex (NRL) allergy has become an important occupational health problem for health care workers, of whom approximately 10% are reportedly sensitized. Some medical facilities have chosen to convert entirely to NRL-free gloves, but others have found this to be financially or logistically unfeasible. OBJECTIVES: The goals of this study were 1) to devise a NRL allergy screening program that could identify sensitized (at-risk) employees for the purpose of providing them with a safe working environment; and 2) to develop a glove conversion plan that would lead to the removal of all NRL gloves from the institution. METHODS: A multidisciplinary University of Maryland Medical System NRL risk team developed a mandatory NRL screening program for all newly hired employees and any existing employees transferring into or within patient care positions. Employees were screened with a clinical questionnaire and a serology for immunoglobulin (Ig)E anti-NRL. Some employees voluntarily received an experimental NRL skin test to evaluate their skin reactivity to NRL. Each employee reporting respiratory or systemic symptoms associated with NRL exposure was evaluated by an allergist. RESULTS: During the 15-month study period from April 1998 to July 1999, 1,795 employees were screened for NRL allergy. Of the whole group, 8% (144 of 1,795) were NRL-specific IgE antibody-positive by CAP radioallergosorbent test (Pharmacia-Upjohn Diagnostics, Kalamazoo, MI) and/or NRL skin test, 57.3% of whom reported symptoms with powdered NRL glove exposure. The NRL IgE-positive rates of nonpatient-care employees (who did not use NRL gloves on a regular basis) and direct patient-care employees was 5.9% and 8.6%, respectively. Most NRL-sensitized employees were successfully accommodated with the use of synthetic gloves and continued to work in the positions for which they were hired. CONCLUSIONS: The University of Maryland Medical System NRL screening program has been successful in identifying at-risk employees and ensuring them a safe working environment during a progressive conversion of the hospital to a NRL safe environment. PMID- 12086363 TI - IgE-mediated natural rubber latex allergy: practical considerations for health care workers. AB - OBJECTIVE: To focus on some specific topics of major significance for health care workers confronted with natural rubber latex (NRL) allergy: 1) sensitization and routes of exposure; 2) threshold allergen exposure levels; 3) evaluation of occupational NRL allergy; 4) prevention of sensitization and allergic reactions to NRL-containing devices; 5) selecting the right alternative; and 6) regulatory responses to NRL allergy issues. DATA SOURCES: English and French language papers identified through a MEDLINE search and bibliographies of the identified papers and the National Institute for Occupational Safety and Health and Food and Drug Administration web sites. RESULTS: 1) The threshold levels of NRL exposure needed for sensitization and the precise way in which it comes about remain to be elucidated, but cutaneous, percutaneous, mucosal, and parenteral exposure can all give rise to symptoms. 2) Strengths and weaknesses of the questionnaire and currently available diagnostic techniques for NRL allergy are emphasized. 3) Prevention strategies should focus on dipped materials and stress upon the possibility of passive and active transmission of NRL aeroallergen. 4) Generally, vinyl gloves are an appropriate alternative for NRL gloves; however, when barrier integrity is a concern, nitrile gloves provide protection that is comparable with NRL. 5) Several government bodies, standards organizations, and regulatory agencies have issued regulations concerning the control and labeling of protein and allergen levels in NRL products. CONCLUSION: Accurate diagnosis and management of NRL allergy is essential because of the potential for severe hypersensitivity reactions. Major developments have been made in characterizing and cloning NRL allergens, and future development in this area may lead to better diagnostic tools and possible therapeutic agents for immunotherapy. However, at present, avoidance remains the only effective approach. We hope that additional well performed prospective incidence studies will bring more accurate data concerning sensitization doses and provide the valuable information to improve prevention strategies. PMID- 12086364 TI - Dust-mite allergen removal from feathers by commercial processing. AB - BACKGROUND: Individuals with allergies or asthma have had a long-standing bias against the use of chicken, goose, and duck feather pillows, comforters, quilts, and mattresses. OBJECTIVE: We show that raw, unprocessed feathers contain dust mite allergen that is removed by washing, and that feather pillows, whether covered or not, do not internally accumulate dust-mite allergen when used in mite infested bedrooms over a 3-month period. METHODS: Feathers obtained from six companies as unprocessed feathers (n = 8 batches) and processed feathers (n = 16 batches) were analyzed for dust-mite (Der p 1 and Der f 1) using monoclonal antibody-based immunoenzyametric assays with appropriate internal controls. Five pairs of new feather pillows (one covered and one uncovered) were placed in five bedrooms with known dust-mite contamination for 90 days. The pillows were then assayed for mite content. RESULTS: Dust-mite allergen was detected in 7 of 8 unprocessed feather samples; combined Der p 1 and Der f 1 mean = 524 ng/g, range 152 to 1,850 ng/g, whereas all 17 manufacturer-processed feather samples contained no detectable dust-mite. Although 4 of 5 bedrooms contained significant dust-mite allergen (>2,000 ng or Der p 1 and f 1, range 2,500 to 10,300 ng/g), none of the 90-day feather pillows became contaminated by dust-mite allergen even in the absence of a pillow cover. CONCLUSIONS: Feathers washed using industrial methods do not contain detectable dust-mite allergen. Pillows manufactured with processed feathers, whether encased in a dust cover or not, do not become contaminated by dust-mite allergen after 90 days of use in environments containing significant dust-mite allergen contamination. PMID- 12086365 TI - Effects of laundry detergents on Dermatophagoides farinae, Dermatophagoides pteronyssinus, and Euroglyphus maynei. AB - BACKGROUND: House-dust mites in clothing and bedding are the source of major allergens. Based on studies of Dermatophagoides pteronyssinus only, weekly washing in hot water is recommended to kill dust mites and remove allergens from clothing and bedding. However, in the United States, washing is most often done in warm or cold water, and other mite species are involved. OBJECTIVE: The purpose of this study was to investigate the lethal effects of various temperatures of hot water alone and hot, warm, and cold water containing detergents and chlorine bleach on Dermatophagoidesfarinae, D. pteronyssinus, and Euroglyphus maynei. METHODS: Mites were soaked in test solutions at various temperatures and for various lengths of time, allowed time to recover, and then analyzed for survival. RESULTS: D. farinae was the most temperature-sensitive and chlorine bleach-sensitive of the three species. In 50 degrees C water alone, 100% mortality for D. farinae was obtained in 10 minutes, whereas most D. pteronyssinus and E. maynei survived. However, 53 degrees C-soaks for 12 and 5 minutes were needed to kill all D. pteronyssinus and E. maynei, respectively. Laundry detergents at their recommended and doubled concentrations and chlorine bleach generally increased mite mortalities over water alone for the three species. Soaking for 4 hours in warm water containing various detergents alone induced mortalities of 19 to 50%, 2 to 35%, and 14 to 46% for D. farinae, D. pteronyssinus, and E. maynei, respectively. CONCLUSIONS: Washing bed linens weekly in warm water with a 4-hour presoak containing most detergents and bleach will kill most D. farinae and, depending on the detergent brand, moderate numbers of D. pteronyssinus. Four-hour soaks in warm water containing the recommended concentrations of various detergents alone also kills moderate numbers of D. farinae, D. pteronyssinus, and E. maynei. Therefore, the cumulative effect of weekly washing with long presoaks should significantly reduce mite levels over time in bed linens, particularly when mattresses and pillows are encased to prevent reinfestation. PMID- 12086366 TI - Therapeutic effects of BCG vaccination in adult asthmatic patients: a randomized, controlled trial. AB - BACKGROUND: Bacille Calmette-Guerin (BCG) vaccination in humans induces Th1 immune responses. Th1 and Th2 cells are reciprocally regulated. OBJECTIVE: To examine whether BCG vaccination of adult patients with asthma, a Th2-associated allergic disease, is clinically effective. METHODS: Forty-three moderate-to severe asthma patients were randomly assigned into groups that received percutaneous injection of 58.2 x 10(7) CFUs BCG (n = 22) or placebo (n = 21) in a double-blinded fashion, on the first day of a 12-week treatment period. Medications were adjusted every 4 weeks to maintain optimal asthma control. Spirometric measurements were performed before treatment and at weeks 4, 8, and 12 after vaccination. The daily peak expiratory flow rate values, asthma symptoms, and medications were also recorded. Tuberculin skin tests, and sputum inflammatory cell and cytokine analyses were carried out before treatment and 12 weeks after vaccination. RESULTS: BCG vaccination significantly increased forced expiratory volume in 1 second and forced expiratory flow rate 25% to 75% at weeks 4, 8, and 12. Morning peak expiratory flow rate was significantly increased only during the first 4 weeks. Although the asthma symptom scores were not significantly changed, the weekly medication scores were significantly decreased. Tuberculin skin reactivities were significantly increased without significant alterations in induced sputum profiles. In contrast, medication scores and sputum eosinophils were significantly increased, and the interferon-gamma:interleukin-4 ratio in sputum was significantly decreased in the placebo group. CONCLUSIONS: BCG vaccination improved lung function and reduced medication use in adults with moderate-to-severe asthma. This amelioration was accompanied by a suppressed Th2 type immune response, suggesting that BCG vaccination might be an effective therapeutic modality against asthma. PMID- 12086367 TI - Efficacy and tolerability of montelukast alone or in combination with loratadine in seasonal allergic rhinitis: a multicenter, randomized, double-blind, placebo controlled trial performed in the fall. AB - BACKGROUND: Histamine and cysteinyl leukotrienes seem to be important mediators of allergic rhinitis. OBJECTIVE: This multicenter, randomized, double-blind, parallel-group, placebo-controlled trial evaluated the effectiveness and tolerability of montelukast, loratadine, and combination therapy with montelukast and loratadine for treating patients with fall seasonal allergic rhinitis. METHODS: After a 1-week, single-blind, placebo run-in period, 907 male and female patients aged 15 to 82 years were randomized to 1 of 4 treatments: montelukast 10 mg (n = 155), loratadine 10 mg (n = 301), combination montelukast 10 mg and loratadine 10 mg (n = 302), or placebo (n = 149), administered once daily at bedtime for 2 weeks. The primary endpoint was the daytime nasal symptoms score (mean of congestion, rhinorrhea, pruritus, and sneezing). RESULTS: Mean symptom scores at baseline were similar for the four treatment groups. For each of the three active treatments, the difference was significant for the mean change from baseline compared with placebo (P < or = 0.001). However, the effect of montelukast/loratadine compared with loratadine alone, the primary comparison, was not significantly different. Differences for each therapy alone compared with placebo were also significant for most secondary endpoints, including nighttime symptom scores, eye symptoms scores, and rhinitis-specific quality of life. Differences for montelukast/loratadine compared with each therapy alone generally showed numerical superiority, and a few endpoints showed differences that were statistically significant. All active treatments showed a safety profile generally similar to placebo. CONCLUSIONS: Montelukast alone or in combination with loratadine is well tolerated and provides clinical and quality-of-life benefits for patients with seasonal allergic rhinitis. PMID- 12086369 TI - Budesonide fatty-acid esterification: a novel mechanism prolonging binding to airway tissue. Review of available data. AB - AIMS: Evidence is accumulating that budesonide (BUD) forms intracellular esters in airways. which may affect both duration of action and therapeutic ratio of this drug. The aim of the present paper is to review the preclinical and human experimental evidence supporting the esterification of BUD, and to discuss the clinical implications this may have on asthma and rhinitis treatment. RESULTS: After inhalation, intact BUD binds primarily to available steroid receptors, and mainly excess (unbound) BUD is esterified. Esterification of BUD is a rapid process: within 20 minutes of inhalation in the rat of radiolabeled BUD, approximately 80% of radioactivity within the trachea and main bronchi was associated with BUD esters, primarily BUD oleate. After 4 hours, the proportion of BUD esters/total cellular BUD was typically 40 to 50% for lung, 70 to 90% for trachea, and only 10 to 15% for peripheral muscle. Comparative in vitro and in vivo studies have shown that esterification prolongs BUD's anti-inflammatory activity longer than that of corticosteroids that can not form esters. Clinical studies have confirmed the prolonged presence of BUD esters, as well as intact BUD, in human airway tissues: 6 hours postdosing, nasal biopsy concentrations of intact BUD were >10-fold greater than those of fluticasone propionate and at 24 hours BUD was detectable in threefold more biopsies than fluticasone propionate. The impact of esterification on airway selectivity of BUD has also been demonstrated in vivo and using pharmacokinetic/pharmacodynamic models. CONCLUSIONS: BUD is retained in airways as esters, a novel kinetic mechanism for synthetic glucocorticoids. In preclinical studies this esterification is correlated to a prolonged local tissue binding and efficacy, which is not found when the esterification is inhibited by an esterification blocker. Because less esters are formed in the systemic compartment than in airways/lung, the local benefit:systemic risk ratio may also be improved by this mechanism. BUD possesses favorable clinical properties, including its approved once-daily efficacy in asthma, which is probably in part attributable to esterification. However, a direct proof of the latter in patients requires effective and safe inhibitors of the esterification, which are not yet available. Therefore, evidence to support the therapeutic impact of esterification is still circumstantial. PMID- 12086368 TI - Multidimensional voice program analysis (MDVP) and the diagnosis of pediatric vocal cord dysfunction. AB - BACKGROUND: Vocal cord dysfunction (VCD) can present with signs and symptoms that mimic asthma. This may lead to unnecessary pharmacologic treatment or more invasive measures including intubation. Presently, the diagnosis of VCD can only be confirmed when a patient is symptomatic, via pulmonary function testing (PFT) or visualization of adduction of the vocal cords during inspiration by direct laryngoscopy. OBJECTIVE: Multidimensional Voice Program (MDVP) analysis. a computer program which analyzes various aspects of voice, can detect abnormal voice patterns of patients with upper airway pathology. We determined whether MDVP analysis was useful in the diagnosis of VCD. METHODS: We conducted chart reviews of patients referred to our department from 1995 to 1998 with the presumed diagnosis of VCD who had undergone MDVP analysis. The diagnosis of VCD was based on the presenting history, PFT results, laryngoscopy results, as well as voice evaluation conducted by a speech-language pathologist. We analyzed six consecutive patients referred for this investigation. We delineated common trends in the variables measured on MDVP analysis in VCD patients. and compared these with controls and other vocal cord pathology. RESULTS: Five cases of possible VCD had abnormalities in the MDVP variable of soft phonation index (SPI). All five also had abnormalities in the variation in fundamental frequency (vFo). In one case, MDVP analysis was conducted pre- and posttreatment for VCD, and SPI and vFo both normalized. In a sixth case of possible VCD. the diagnosis was not confirmed as the patient had normal PFTs and laryngoscopy. MDVP analysis was normal in this individual. The pattern of abnormal SPI and vFo was not seen in a group of normal controls or in patients with vocal cord nodules. CONCLUSIONS: MDVP analysis may be a useful tool when diagnosingVCD, as well as in evaluating response to treatment. PMID- 12086370 TI - A randomized comparison of the effects of budesonide and mometasone furoate aqueous nasal sprays on nasal peak flow rate and symptoms in perennial allergic rhinitis. AB - BACKGROUND: Using conventional methods, it has been difficult to show differences in efficacy between intranasal corticosteroids in perennial rhinitis. OBJECTIVE: To compare the effects of budesonide and mometasone on nasal symptoms and nasal airflow in perennial allergic rhinitis. METHODS: Four hundred thirty-eight patients (age > 18 years old) were randomized to budesonide, 256 microg or 128 microg, mometasone furoate 200 microg, or placebo, once daily for 4 weeks. Efficacy was evaluated by nasal index score (NIS; the sum of scores for blocked nose, runny nose, and itchy nose/sneezing) and peak nasal inspiratory flow (PNIF). RESULTS: All three active treatments significantly reduced the NIS compared with placebo. There was no significant difference between the treatments, although the effect of budesonide, 256 microg, tended to be greater than that of the other regimens. PNIF was significantly improved with all three active treatments: the effect of budesonide 256 microg on morning and evening PNIF was significantly greater than that of mometasone furoate and 128 microg budesonide. Budesonide had a rapid onset of action, showing a significantly greater effect on evening PNIF than mometasone furoate during the first 10 days. For all active treatments, significant improvements in NIS were seen within 4 hours of the first dose. All three treatments were well tolerated. CONCLUSION: The objective parameter PNIF was capable of demonstrating greater efficacy of budesonide 256 microg compared with budesonide 128 microg and mometasone furoate 200 microg, whereas the combined nasal symptom score could only distinguish active treatment from placebo. PMID- 12086371 TI - Nasal allergen provocation with Dermatophagoides pteronyssinus in patients with chronic rhinitis referred to a rhinologic surgical center. AB - BACKGROUND: Skin tests and specific immunoglobulin (Ig)E determinations identify the presence of some immune response to allergens, but frequently do not correlate to allergic disease in vivo. OBJECTIVE: To compare clinical history, skin tests and specific IgE determinations with the results of nasal provocation tests (NPT) to confirm clinically relevant allergy to Dermatophagoides pteronyssinus (DP) in patients with chronic rhinitis referred to a rhinologic surgical center. METHODS: Clinical data of 2,194 patients with chronic nasal disorders treated at two rhinologic referral centers between 1992 and 1998 were evaluated. Clinical allergy work-up was routinely performed corresponding to German and European guidelines providing consistent data in both centers. RESULTS: Of chronic rhinitis patients, 752 of 2,194 (34%) had at least one positive skin reaction to a perennial allergen common in Central Europe. Three hundred seventy-three patients had a positive skin test to DP and were paradigmatically evaluated in detail. In 235 patients, a complete dataset including structured allergic history, skin test, specific IgE determinations, and NPT was available. Quality and severity of symptoms, clinical history, skin tests, and allergen-specific IgE-determinations were unreliable predictors of the outcome of NPT with DP. CONCLUSION: In patients referred for surgical treatment of chronic rhinitis, diagnosis of perennial allergy is unreliable unless confirmed with NPT. PMID- 12086372 TI - Polyclonal autoantibodies against C1 inhibitor in a case of acquired angioedema. AB - BACKGROUND: Angioedema attributable to acquired C1 inhibitor (C1-INH) deficiency is a rare disease related to lymphoproliferative disorders or autoantibodies to Cl inhibitor. We describe a patient with angioedema and autoantibodies to C1 inhibitor. OBJECTIVE: To study the characteristics of autoantibodies to C1-INH in a patient with acquired angioedema. METHODS: Autoantibodies to Cl-INH were measured by enzyme-linked immunoadsorbent assay. Immunoglobulin (Ig)G autoantibody was purified by affinity chromatography on a protein G agarose column. We developed an enzyme-linked immunoadsorbent assay to determine whether the autoantibodies were directed against the C1-INH active center. RESULTS: IgM and mainly C1-INH IgG autoantibodies were detected; both had kappa and lambda chains. No monoclonal component was detected. The autoantibodies were directed against the Cl-INH active center. After various treatment strategies were attempted, an effective clinical response was attained with antifibrinolytic therapy. CONCLUSION: A case of acquired angioedema because of C1-INH deficiency was found to be attributable to the presence of polyclonal autoantibodies to C1 INH. PMID- 12086373 TI - Occupational allergy caused by Peruvian lily (Alstroemeria). AB - BACKGROUND: Allergic contact dermatitis to Alstroemeria has been well documented; however, occupational allergy to this decorative flower has never been reported in the literature. OBJECTIVE: We describe a florist with complaints of a sense of throat tightness, allergic rhinoconjunctivitis, urticaria, and facial angioedema attributable to the handling of this popular flower. METHODS: An allergy skin testing by the puncture technique and a challenge test are performed in a private office. A staff member is used as a control for the skin testing. Main outcome measures are the subject's clinical symptoms. RESULTS: The allergy skin testing reveals positive response to Alstroemeria (Peruvian lily), but negative to Stargazer lily, solidago, and few other flower extracts. In the challenge test, the subject develops conjunctival injection, postnasal drip with nasal congestion, and cough. CONCLUSIONS: This is the first report of a type I allergic reaction to Alstroemeria and illustrate the ease of in-office performance of skin testing and challenge to flowering plants. PMID- 12086374 TI - Churg-Strauss syndrome: survival for 26 years. AB - BACKGROUND: Churg-Strauss syndrome (CSS), a necrotizing vasculitis characterized by asthma and eosinophilia, was described initially in 1951. Before the use of oral corticosteroids, the average patient survived for only a few months. Today with the use of oral corticosteroids and immunosuppressants, survival has increased significantly. METHODS: We report the case of a patient with CSS treated with prednisone and azathioprine. A review of the English literature was performed with MEDLINE from 1966 to the present using these keywords: Churg Strauss syndrome, survival, prognosis, morbidity, mortality, and treatment. RESULTS: This patient survived 26 years after the diagnosis of CSS and died without autopsy findings of active vasculitis. This is the longest reported survival with CSS in the English literature to our knowledge. The patient's disease course was marked by two acute (vasculitic) episodes, with intermittent subacute disease, and finally a state of disease remission. CONCLUSIONS: Survival in patients with CSS can be prolonged with early initiation of corticosteroids and immunosuppressants, close outpatient followup, and prompt, aggressive treatment of relapses. This case exemplifies the disease progression and remission as reported by other authors. We propose that CSS can be classified into acute (vasculitic), subacute, and remittable stages. PMID- 12086375 TI - Malignant idiopathic anaphylaxis does exist in Brazil. PMID- 12086376 TI - Takayasu's arteritis revisited. PMID- 12086377 TI - Congenital long Q-T syndromes in children. PMID- 12086378 TI - Beating heart versus conventional reoperative coronary artery bypass surgery. AB - BACKGROUND: The incidence of reoperative coronary artery bypass grafting is increasing with an increase in the number of patients undergoing coronary artery bypass surgery. The clinical outcome of redo coronary artery bypass grafting without cardiopulmonary bypass and conventional coronary artery bypass grafting using cardiopulmonary bypass are different. METHODS AND RESULTS: We compared clinical parameters in patients who underwent off-pump (n=156) versus on-pump (n=194) redo coronary artery bypass grafting performed between January 1995 and December 2001 in our institute, to determine if off-pump surgery has improved the surgical outcome of redo coronary artery bypass grafting and emerged as an ideal technique. Patients who underwent on-pump redo surgery required more postoperative blood transfusion (86.53% on-pump v. 12.82% off-pump. p=0.001), prolonged ventilatory support (>24 hours) (16.49% on-pump v. 7.7% off-pump, p=0.021) and higher inotropic support (23.71% on-pump v. 10.89% off-pump, p=0.003). On-pump redo coronary artery bypass grafting was also associated with a prolonged stay in the intensive care unit (40+/-6.2 hours on-pump v. 20+/-4.1 hours off-pump, p=0.001) and longer hospital stay (9+/-4.2 days on-pump v. 5+/ 3.4 days off-pump, p=0.001). In-hospital mortality was higher in on-pump patients than in off-pump ones (7.7% v. 3.2%); however, this was not statistically significant (p=0.114). CONCLUSIONS: Off-pump redo coronary artery bypass grafting is a safe method of myocardial revascularization with lower operative morbidity and mortality, less requirement of blood products and early hospital discharge, compared with conventional on-pump redo coronary artery bypass grafting. PMID- 12086379 TI - Graded balloon atrial septostomy in severe pulmonary hypertension. AB - BACKGROUND: The prognosis of patients with severe primary pulmonary hypertension is poor. The role of balloon atrial septostomy as a palliative procedure in these patients is not well defined. We retrospectively analyzed our data regarding the safety, clinical outcome and survival benefit of graded balloon atrial septostomy in patients with severe pulmonary hypertension. METHODS AND RESULTS: Eleven patients (7 males), aged 6 to 30 years (mean age 16.2+/-8.9 years), with severe pulmonary artery hypertension (mean pulmonary artery pressure of 76+/-16.9 mmHg) and refractory congestive heart failure and/or recurrent syncope underwent balloon atrial septostomy. Graded balloon dilatation under echocardiographic guidance and arterial oxygen saturation monitoring was done in all the patients. Procedure-related mortality was 18.2%. Significant acute hemodynamic improvement was seen in the survivors (pre-balloon atrial septostomy cardiac index 1.88+/ 0.48 L/min/m2; post-balloon atrial septostomy cardiac index 2.18+/-0.37 L/min/m2, p<0.009). Patients were followed up for a mean period of 20.3 months after the procedure (range: 3 months-5 years). There was functional improvement and increased exercise tolerance in all the patients for a mean follow-up period of 14.6 months (NYHA functional class 3.62+/-0.69 to 2+/-0.50). The estimated probability of survival in this cohort at 1 year was only 48%; but 7 of 8 patients (87%) who survived the procedure were alive at 1 year. CONCLUSION: We conclude that balloon atrial septostomy improves clinical status, hemodynamic variables and possibly also improves survival in selected patients with severe pulmonary artery hypertension. It remains a definite palliative option for refractory primary pulmonary hypertension. However, the procedure-related risks are high in very sick patients and, therefore, balloon atrial septostomy may be advocated early in the course of the disease. PMID- 12086380 TI - Efficacy of Terminalia arjuna in chronic stable angina: a double-blind, placebo controlled, crossover study comparing Terminalia arjuna with isosorbide mononitrate. AB - BACKGROUND: Terminalia arjuna, an Indian medicinal plant, has been reported to have beneficial effects in patients with ischemic heart disease in a number of small, open studies. The need for a double-blind, randomized, placebo-controlled study with adequate sample size has long been felt. The bark extract (IPC-53) contains acids (arjunic acid, terminic acid), glycosides (arjunetin arjunosides I IV), strong antioxidants (flavones, tannins, oligomeric proanthocyanidins), minerals. etc. and exhibits antifailure and anti-ischemic properties. METHODS AND RESULTS: Fifty-eight males with chronic stable angina (NYHA class II-III) with evidence of provocable ischemia on treadmill exercise test received Terminalia arjuna (500 mg 8 hourly), isosorbide mononitrate (40 mg/daily) or a matching placebo for one week each, separated by a wash-out period of at least three days in a randomized, double-blind, crossover design. They underwent clinical, biochemical and treadmill exercise evaluation at the end of each therapy which were compared during the three therapy periods. Terminalia arjuna therapy was associated with significant decrease in the frequency of angina and need for isosorbide dinitrate (5.69+/-6.91 mg/week v. 18.22+/-9.29 mg/week during placebo therapy, p<0.005). The treadmill exercise test parameters improved significantly during therapy with Terminalia arjuna compared to those with placebo. The total duration of exercise increased (6.14+/-2.51 min v. 4.76+/-2.38 min, p<0.005), maximal ST depression during the longest equivalent stages of submaximal exercise decreased (1.41+/-0.55 mm v. 2.21+/-0.56 mm, p<0.005), time to recovery decreased (6.49+/-2.37 min v. 9.27+/-3.39 min, p<0.005) and higher double products were achieved (25.75+/-4.81x10(3) v. 23.11+/-4.83x10(3), p<0.005) during Terminalia arjuna therapy. Similar improvements in clinical and treadmill exercise test parameters were observed with isosorbide mononitrate compared to placebo therapy. No significant differences were observed in clinical or treadmill exercise test parameters when Terminalia arjuna and isosorbide mononitrate therapies were compared. No significant untoward effects were reported during Terminalia arjuna therapy. CONCLUSIONS: Terminalia arjuna bark extract, 500 mg 8 hourly, given to patients with stable angina with provocable ischemia on treadmill exercise, led to improvement in clinical and treadmill exercise parameters as compared to placebo therapy. These benefits were similar to those observed with isosorbide mononitrate (40 mg/day) therapy and the extract was well tolerated. Limitations of this study include applicability of the results to only men with chronic stable angina but not necessarily to women, as they were not studied. PMID- 12086381 TI - Management of cardiomyopathy resulting from incessant supraventricular tachycardia in infants and children. AB - BACKGROUND: Radiofrequency ablation is considered to be the treatment of choice in patients with ventricular dysfunction related to incessant supraventricular tachycardia. However, reservations regarding its use in infants and children prompted us to try alternative strategies for this group. METHODS AND RESULTS: Eight children (age range: 1 day to 10 years) were diagnosed to have tachycardia related ventricular dysfunction in the past 6 years. They presented with symptoms of palpitation, dyspnea and/or generalized swelling over the body of 3 months to 2 years'duration. The cardiothoracic ratio at presentation was 64% (52%-70%) and ejection fraction was 22.2% (15%-45%). In 7 patients tachycardia was diagnosed to be ectopic atrial and in 1 it was permanent junctional reciprocating tachycardia. Six of these children were managed with intravenous/oral amiodarone in combination with digoxin (3) and/or propranolol (2). In one child addition of amiodarone to digoxin and propranolol led to polymorphic ventricular tachycardia, and amiodarone was withdrawn. Only one child underwent radiofrequency ablation as the first choice because regular follow-up was not possible due to logistic reasons. Sinus rhythm with normalization of ventricular function was achieved in 6 of the 7 children treated medically. One child continued to have frequent episodes of tachycardia and underwent successful radiofrequency ablation of a high right atrial ectopic focus. Two out of the 6 patients on amiodarone could be managed with only digoxin and propranolol after their ventricular function had returned to normal. A third patient relapsed on stopping amiodarone and underwent successful radiofrequency ablation of a left atrial ectopic tachycardia. CONCLUSIONS: Short-term amiodarone in combination with digoxin/propranolol is a safe and effective treatment strategy for infants/children with tachycardiomyopathy. Control of tachycardia is achieved in the majority, leading to recovery of ventricular function. This approach may avoid unnecessary ablations in children or at least postpone it till the procedure would be safer. PMID- 12086382 TI - Prospective evaluation of the risk of bacteremia induced by transesophageal echocardiography. AB - BACKGROUND: The incidence of bacteremia induced by transesophageal echocardiography is controversial in the Indian population. This study aimed to find out the occurrence of bacteremia following transesophageal echocardiography. METHODS AND RESULTS: Between February 2000 and January 2001, 47 patients (26 males and 21 females) were enrolled for the study. Their ages ranged from 13 to 61 years (mean: 35 +/- 11.4 years). Patients with prosthetic valves, suspected infective endocarditis and those on antibiotics were excluded. For each procedure, two sets of blood cultures were obtained immediately before and after the procedure. For each blood culture, 10 ml of blood was evenly inoculated into brain-heart infusion broth and biphasic infusion medium and incubated for 7 days. Transesophageal echocardiography was carried out under oropharyngeal anesthesia (xylocaine gel and spray). Two blood cultures taken before the procedure were positive and excluded from the final analysis. Of the remaining 45 patients whose preprocedure blood cultures were sterile, 6 samples (13.3%) were positive after the procedure diphtheroids in 3, micrococci in 2 and aerobic spore formers in 1. CONCLUSIONS: This study demonstrates that the incidence of bacteremia related to transesophageal echocardiography is not insignificant, as reported in previous studies. Though routine antibiotic prophylaxis before transesophageal echocardiography is not advocated, it should be recommended in high-risk patients such as those with prosthetic valves, multivalvular involvement or those with a past history of infective endocarditis. PMID- 12086384 TI - Bioptome-assisted coil occlusion of coronary artery fistula. AB - We describe a novel technique that allows controlled and precise delivery of single or multiple coils simultaneously for occlusion of a coronary artery fistula using a bioptome passed via a long sheath positioned at the distal end of the fistula. The fistula was balloon occluded distal to the take-off of the native branches before, during and after coil delivery in two patients. PMID- 12086383 TI - Coronary angiography using 4 French catheters with power injection: a randomized comparison with 6 French catheters. AB - BACKGROUND: Coronary angiography using 4 F catheters may reduce access-site complications and enable early ambulation, although earlier studies suggested that the quality of images may be an issue of concern. METHODS AND RESULTS: To ascertain the quality of angiographic images and safety of early ambulation, 500 patients were randomized to coronary angiography with either 4 F or 6 F catheters. Procedural characteristics, angiographic quality scores and results of ambulation were analyzed in the two groups. Patients in the 4 F group were mobilized at 2 hours post-procedure while those in the 6 F group were ambulated at 6 hours. There was no procedure-related complication in either group. The procedure was successfully completed in 250 of 252 patients randomized to the 4 F group. In two patients in the 4 F group, sheaths were upgraded to 6 F to complete the procedure, as difficulty was encountered in hooking the coronary ostium with a 4 F Judkin's catheter. Coronary angiographic quality scores in these two groups were comparable. Angiographic scores for the 4 F and 6 F groups for the left coronary artery averaged 4.45+/-0.5 and 4.58+/-0.3 (p>0.1), respectively. The right coronary artery scores averaged 4.30+/-0.4 and 4.35+/-0.2 (p>0.1) in the 4 F and 6 F groups. Angiographic scores for the left ventricular angiogram averaged 4.22+/-0.1 and 4.44+/-0.3 (p>0.1) in the 4 F and 6 F groups, respectively. None of the angiograms were assigned a score of <3.0 (not diagnostic). The total contrast volume consumed in the two groups was also equivalent. There were no groin-related complications in the 4 F group although these patients were ambulated 2 hours after the procedure. CONCLUSIONS: Coronary angiography performed with a 4 F catheter is a safe and reliable procedure. The quality of image obtained with a 4 F catheter is equivalent to that obtained with a 6 F catheter. Early ambulation at 2 hours is feasible without compromising safety. PMID- 12086385 TI - Familial primary pulmonary hypertension. AB - Primary pulmonary hypertension is a rare disease affecting mostly females. We report a family where 2 of the 3 male children born to consanguineous parents had severe pulmonary hypertension of unexplained cause. The occurrence of overtly manifest primary pulmonary hypertension is rare in males, especially at an early age. PMID- 12086386 TI - Left ventricular thrombi in the presence of normal left ventricular function. AB - We report two cases of left ventricular thrombi identified by routine echocardiography in the presence of normal ventricular function to highlight the rarity and clinical significance of this condition. A 14-year-old boy, positive for anticardiolipin and antinuclear antibodies, was found to have a left ventricular thrombus. A 30-year-old male, who presented with a transient ischemic attack, was found to have hypereosinophilic syndrome and a mobile left ventricular thrombus. The thrombi disappeared in both patients after a few days of anticoagulant therapy without symptoms of embolization. PMID- 12086387 TI - Thromboembolism: a rare complication of cardiac hydatidosis. AB - A cardiac hydatid cyst is rare. We report a case of cardiac hydatid cyst localized in the atria which was diagnosed by two-dimensional echocardiography following a thromboembolic stroke. Surgical resection of the cyst was performed and histopathologic examination confirmed the diagnosis. PMID- 12086389 TI - Reduction of pacemaker-induced pectoral muscle stimulation using an insulating patch. AB - An 82-year-old female patient was admitted for replacement of her pulse generator battery which had reached an end-of-life situation. The old unipolar leads were retained. Pectoral muscle stimulation began shortly after implantation. Though complete abolition of muscle stimulation was not achieved, an insulating patch placed between the muscle and the pacemaker can reduced stimulation. PMID- 12086388 TI - Extrinsic obstruction of the main coronary arteries in a child with hypertrophic cardiomyopathy. AB - Phasic narrowing of the coronary arteries on angiography is a well-known entity in both children and adults and has been described in relation to all epicardial arteries. There is a high incidence of myocardial bridges in hypertrophic cardiomyopathy. We report the case of a 6-year-old girl with hypertrophic obstructive cardiomyopathy who had extrinsic obstruction of the left main and right coronary arteries. PMID- 12086390 TI - Apical hypertrophic cardiomyopathy in association with Takayasu's arteritis. AB - We describe an interesting case of a patient who had Takayasu's arteritis and apical hypertrophic cardiomyopathy. Electrocardiogram, and transthoracic and transesophageal echocardiograms showed classical features of apical hypertrophic cardiomyopathy which is particularly uncommon outside Japan. To the best of our knowledge, the presence of apical hypertrophic cardiomyopathy in patients with Takayasu's arteritis has not been reported till date. PMID- 12086391 TI - Drug-eluting stents in the treatment of atherosclerotic coronary heart disease. PMID- 12086392 TI - Loop the loop. PMID- 12086393 TI - Clopidogrel and cardiac surgery. PMID- 12086394 TI - Prognostic significance of rapid bedside cardiac troponin T testing in unstable angina. PMID- 12086395 TI - Is hypertrophic obstructive cardiomyopathy really different in elderly patients? A case of typical HOCM detected in a 94-year-old female. PMID- 12086396 TI - The role of nitric oxide and cyclooxygenase-2 in attenuating apoptosis. AB - The production of nitric oxide (NO) is an essential determinant in auto- and paracrine signaling. NO is generated under inflammatory conditions and may serve as a cytotoxic molecule to produce cell demise along an apoptotic or necrotic pathway. NO also gained attention as a regulator of immune function and a death inhibitor. Cytotoxicity because of substantial NO-formation is established to initiate apoptosis, characterized by upregulation of the tumor suppressor p53, changes in the expression of pro- and antiapoptotic Bcl-2 family members, cytochrome c relocation, activation of caspases, and DNA fragmentation. However, NO-toxicity is not a constant value and NO may protect several cell types from entering programmed cell death. Preactivation of macrophages with a nontoxic dose of S-nitrosoglutathione (200 microM) or lipopolysaccharide/interferon-gamma/N(G) monomethyl-L-arginine for 15 hours attenuated death in response to various agonists, suppressed p53 accumulation, and abrogated caspase activation. Prestimulation of macrophages with cytokines or low-level NO activated the transcription factor NF-kappaB as well as AP-1 and promoted immediate early gene expression of cyclooxygenase-2 (COX-2). NF-kappaB activation comprised p50/p65 heterodimer formation, IkappaB degradation, and activation of a luciferase reporter construct, that contained four copies of the NF-kappaB-site derived from the murine COX-2 promoter. A NF-kappaB decoy approach (oligonucleotides directed against NF-kappaB) or transfection of a dominant-negative c-Jun mutant (TAM67) abrogated not only the COX-2 expression but also the inducible protection. Blocking NO- or cytokine-mediated inducible protection at the level of NF-kappaB and/or AP-1 restored the occurrence of apoptotic features. Our experiments underscore the role of COX-2 in attenuating natural occurring cell death (i.e., apoptosis). PMID- 12086397 TI - Redox status-dependent regulation of cyclooxygenases mediates the capsaicin induced apoptosis in human neuroblastoma cells. AB - Cyclooxygenases (COX) appear to be involved in the mechanism of apoptosis in various cancer cells. In this study we investigated the role of COX in the capsaicin (Cap)-induced apoptosis in SK-N-SH human neuroblastoma cells. Cap induced decreased cell viability and apoptosis in a dose-dependent manner. Cap also significantly reduced the basal generation of reactive oxygen species (ROS) and lipid peroxidation in a time-dependent fashion. Cap markedly suppressed the expression of COX-1 and COX-2. Pretreatment with NS-398, a selective COX-2 inhibitor, or indomethacin, a nonselective COX inhibitor, significantly enhanced the Cap-induced decreased cell viability and apoptosis. Exogenous application of an oxidant, H2O2, significantly prevented the Cap-induced apoptosis and suppressed the expression of COX isoforms. These results suggest that redox status-dependent regulation of COX expression may mediate apoptosis induced by Cap in human neuroblastoma cells. PMID- 12086398 TI - Oxidant-sensitive transcription factor and cyclooxygenase-2 by Helicobacter pylori stimulation in human gastric cancer cells. AB - Helicobacter pylori (H. pylori) infection might activate nuclear factor-kappaB (NF-kappaB), an oxidant-sensitive transcription regulator of inducible expression of inflammatory genes such as cyclooxygenase-2 (COX-2). We studied the role of NF kappaB on expression of COX-2 in H. pylori-stimulated gastric cancer cell lines by using antioxidants, glutathione (GSH), and N-acetylcysteine (NAC) as well as an NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC). Gastric adenocarcinoma cell lines derived from Caucasian (AGS) cells and Korean (SNU-484) cells were used to study the role of NF-kappaB on COX-2 expression by H. pylori. They were treated with GSH, NAC, or PDTC in the presence of H. pylori. mRNA expression and protein level for COX-2 were determined by Northern blot and RT PCR analysis as well as Western blot analysis. NF-kappaB activation was examined by electrophoretic mobility shift assay. As a result, H. pylori induced a time dependent expression of mRNA and protein for COX-2 via activation of NF-kappaB, which was inhibited by GSH, NAC, and PDTC in the cells. In conclusion, oxidant sensitive transcription factor NF-kappaB may play a novel role in expression of COX-2 by H. pylori stimulation in gastric cancer cells. PMID- 12086399 TI - Effects of yakuchinone A and yakuchinone B on the phorbol ester-induced expression of COX-2 and iNOS and activation of NF-kappaB in mouse skin. AB - Certain medicinal plants contain anti-inflammatory and antioxidative substances that can exert chemopreventive effects. Our previous studies have demonstrated that the methanol extract of Alpinia oxyphylla Miquel (Zingiberaceae) inhibits tumor promotion in mouse skin. Two major diarylheptanoids named yakuchinone A (1 [4'-hydroxy-3'-methoxyphenyl]-7-phenyl-3-heptanone) andyakuchinone B (1-[4' hydroxy-3'-methoxyphenyl]-7-phenylhept-1-en-3-one) have been isolated from this medicinal plant. Both compounds have strong inhibitory effects on the synthesis of prostaglandins and leukotrienes in vitro. In the present work, we show that both yakuchinone A and yakuchinone B inhibit the expression of cyclooxygenase-2 (COX-2) and of inducible nitric oxide synthase (iNOS) as well as the expression of tumor necrosis factor (TNF)-alpha mRNA in mouse skin treated with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). Topical application on mouse skin of these diarylheptanoids also attenuated the TPA-induced DNA binding activity of the ubiquitous eukaryotic transcription factor NF-kappaB that plays a crucial role in regulating the expression of the aforementioned proinflammatory enzymes and cytokines in response to a wide variety of external stimuli. These findings suggest that diarylheptanoids contained in Alpinia oxyphylla down regulate COX-2 and iNOS expression through suppression of NF-kappaB activation in the TPA-treated mouse skin. PMID- 12086400 TI - Suppressive effect of natural sesquiterpenoids on inducible cyclooxygenase (COX 2) and nitric oxide synthase (iNOS) activity in mouse macrophage cells. AB - Prostaglandins and nitric oxide produced by inducible cyclooygenase (COX-2) and nitric oxide synthase (iNOS), respectively, have been implicated as important mediators in the processes of inflammation and carcinogenesis. These potential COX-2 and iNOS inhibitors have been considered as antiinflammatory and cancer chemopreventive agents. In this study, we investigated the effect of natural sesquiterpenoids isolated from plants of the Zingiberaceae family on the activities of COX-2 and iNOS in cultured lipopolysaccharide (LPS)-activated mouse macrophage cell RAW 264.7 to discover new lead compounds as COX-2 or iNOS inhibitors. Xanthorrhizol, a sesquiterpenoid, isolated from the rhizome of Curcuma xanthorrhiza Roxb. (Zingiberaceae), exhibited a potent inhibition of COX 2 (IC50 = 0.2 microg/mL) and iNOS activity (IC50 = 1.0 microg/mL) in the assay system of prostaglandin E2 (PGE2) accumulation and nitric oxide production, respectively. Western blot analyses revealed that the inhibitory potential of xanthorrhizol on the COX-2 activity coincided well with the suppression of COX-2 protein expression in LPS-induced macrophages. In addition, sesquiterpenoids beta turmerone and ar-turmerone isolated from the rhizome of Curcuma zedoaria Roscoe (Zingiberaceae) also showed a potent inhibitory activity of COX-2 (beta turmerone, IC50 = 1.6 microg/mL; ar-turmerone, IC50 = 5.2 microg/mL) and iNOS (beta-turmerone, IC50 = 4.6 microg/mL; ar-turmerone, IC50 = 3.2 microg/mL). These results suggest that natural sesquiterpenoids from C. xanthorrhiza and C. zedoaria might be lead candidates for further developing COX-2 or iNOS inhibitors possessing cancer chemopreventive or anti-inflammatory properties. PMID- 12086401 TI - COX-2 and prostanoid receptors: good targets for chemoprevention. AB - Accumulating evidence indicates that COX-2 inhibitors are involved in colon and breast cancer development. Our previous studies indicated that nimesulide and celecoxib, selective COX-2 inhibitors, show inhibitory effects of intestinal carcinogenesis in azoxymethane-treated rats and mice and in Min mice models. We recently found that nimesulide suppressed PhIP-induced breast cancer in female SD rats in which COX-2 protein was overexpressed. These results led us to investigate the effects of prostaglandin E2 (PGE2) in the target tissues. PGE2 showed its biological activity through binding to its membrane receptors, EP(1 to approximately 4). We also investigated the effects of EP receptors on colon carcinogenesis. We used receptor knockout mice and selective receptor antagonists. Our results indicated that the EP1 receptor plays a pivotal role in colon carcinogenesis. Selective EP1 receptor antagonists may be a new class of chemopreventive agents against colon cancer. PMID- 12086402 TI - Novel approaches for colon cancer prevention by cyclooxygenase-2 inhibitors. AB - During recent years, multidisciplinary studies in epidemiology and molecular biology, as well as preclinical studies, have contributed much to our understanding of the etiology of colorectal cancer; more importantly they have enabled us to approach its prevention. An impressive body of epidemiological data suggests an inverse relationship between colorectal cancer risk and regular use of nonsteroidal antiinflammatory drugs (NSAIDs), including aspirin. Clinical trials with NSAIDs have demonstrated that NSAID treatment caused regression of preexisting colon adenomas in patients with familial adenomatous polyposis. Preclinical efficacy studies have provided compelling evidence that several phytochemicals with antiinflammatory properties and NSAIDs act to retard, block, or reverse colon carcinogenesis. Equally exciting are opportunities for effective chemoprevention with selective cyclooxygenase-2 (COX-2) inhibitors including celecoxib and rofecoxib in a variety of preclinical models of colon cancer. Naturally occurring COX-2 inhibitors such as curcumin and certain phytosterols have been proven to be effective as chemopreventive agents against colon carcinogenesis with minimal gastrointestinal toxicity. Multistep process of carcinogenesis has provided substantial insights into the mechanisms by which naturally occurring and synthetic antiinflammatory agents modulate these events leading to suppression of tumorigenesis. Growing knowledge in this area has brought about innovative approaches using a combination of agents with different modes of action as a means of increasing efficacy and minimizing toxicity. The natural history of colorectal cancer, from dysplastic aberrant crypts to adenomas and adenocarcinomas, offers multiple opportunities for assessment and intervention. Of further importance would be to identify molecular targets that are critical in the growth and survival of the malignant colorectal cell and are modulated by NSAIDs and COX-2 inhibitors. PMID- 12086403 TI - In vitro evidence of the role of COX-2 in attenuating gastric inflammation and promoting gastric carcinogenesis. AB - Although gastric adenocarcinoma is one of the most common malignancies in the world, little is known about its exact molecular processes in development and progression. Recent studies suggest that COX-2 is important in carcinogenesis of gastrointestinal cancers, and is especially involved in carcinogenesis in a mouse model of familial adenomatosis polyposis. To understand the role of COX-2 in gastric carcinogenesis and Helicobacter pylori-associated gastritis, we measured COX-2 expression in 170 human gastric carcinoma tissues byimmunohistochemical analysis and compared the expression of COX-2 in paired tissues obtained from normal-looking and cancer-bearing mucosa. Further evidence of the involvement of COX-2 in gastritis and gastric carcinogenesis was obtained by establishing stable cell lines overexpressing COX-2. After subcloning of COX-2 into pCB7 mammalian expression vector, two stable cell lines named MKN-28-COX-2 and MKN-45-COX-2 were generated by transfection of COX-2 cDNA. To understand the effect of COX-2 on gastritis, we performed an electrophoretic mobility shift assay of NF-kappaB (inflammation-associated transcription factor), and measured malondialdehyde levels and chemiluminescence activities in both mock-transfected MKN and MKN-COX 2 cells after stimulation of H. pylori (1 x 10(6) CFU/mL) and neutrophils (10(2) cells/mL). A marked attenuation of NF-kappaB bindings and generation of free radicals was observed in COX-2 overexpressed cells. Another set of experiments, including the growth inhibition by TGF-beta treatment, Matrigel invasion assay, and apoptosis assay, was done. COX-2 showed the advantage of the escape from the growth inhibition by TGF-beta through decreasing TGF-beta RII expression and increased cell invasiveness. In conclusion, COX-2 expression seems to be induced to attenuate the degree of atrophic gastritis, the initial event in gastric carcinogenesis, and promote gastric carcinogenesis. PMID- 12086404 TI - Increased expression of COX-2 in the development of human lung cancers. AB - It is well accepted that an increase in the expression of cyclooxygenase-2 (COX 2), a key inducible enzyme involved in the production of prostaglandins and other eicosanoids, may play a significant role in carcinogenesis in addition to its well-known role in inflammatory reactions. Whereas previous studies were largely confined to colorectal tumorigenesis, we have shown that a significantly increased expression of COX-2 may also play a role in the development of lung cancer. COX-2 expression was found to be frequently elevated in lung cancer, especially in adenocarcinoma, and the proportion of lung cancer cells with marked COX-2 expression was much higher in lymph node metastases than in the corresponding primary tumors. It was also shown that early stage adenocarcinoma patients with increased COX-2 expression who were surgically treated had a shorter survival. Our studies, which used high- and low-metastatic human lung cancer cell sublines established in our laboratory, revealed an association between metastatic capabilities and COX-2 expression levels: COX-2-specific inhibitors could inhibit in vitro the invasion of the highly metastatic NCI-H460 LNM35 clone through Matrigel-containing basement membrane components as well as the spontaneous in vivo metastasis in SCID mice. Taken together, these findings suggest that an increase in COX-2 expression maybe associated with the development of lung cancer and possibly with the acquisition of an invasive and metastatic phenotype. PMID- 12086405 TI - Is cyclooxygenase-2 important in skin carcinogenesis? AB - Our studies have focused on the role of arachidonic acid and its products in chemically and UV light-induced murine models of skin carcinogenesis, with an emphasis on determining the importance of prostaglandins (PGs), which are synthesized by the two isoforms of cyclooxygenase (COX). Different types of tumor promoters elevate COX-2 expression in keratinocytes, with little change in COX-1, suggesting that there are multiple signaling pathways by which COX-2 expression can be regulated. We found that the expression of both COX isoforms is increased by treatment with PGs and that this autoregulation occurs via PG receptors linked to a cAMP signaling pathway. We also observed that COX-2 is constitutively upregulated in papillomas and carcinomas from either chemical initiation promotion or UV-irradiation carcinogenesis experiments. We next investigated cis- and transacting factors required for COX-2 expression. Two regions of the COX-2 promoter, an E box and a nuclear factor-IL6 (NF-IL6) site, were identified as positive regulatory elements through transient transfection with luciferase reporter vectors containing various 5'-flanking regions of the promoter. We found that overexpression of COX-2 in tumors maybe caused by a dysregulation in the expression pattern of the CCAAT/enhancer binding protein (C/EBP) family of transcription factors. To demonstrate the importance of PG synthesis in the carcinogenesis process, several nonsteroidal anti-inflammatory (NSAIDs) drugs were administered either orally or topically during UV carcinogenesis. Dietary administration of indomethacin, piroxicam, or the selective COX-2 inhibitor celecoxib prevented the development of UV-induced skin cancers by up to 85%. In addition, celecoxib had therapeutic efficacy in that it caused regression of preexisting tumors. Topical administration of indomethacin after each UV exposure was also effective, suggesting that a postexposure approach to skin cancer prevention maybe effective. Collectively, these studies suggest that prostaglandins play a critical role in skin cancer development. PMID- 12086406 TI - Epidemiological and clinical aspects of nonsteroidal anti-inflammatory drugs and cancer risks. AB - It is well known that about 70% of cancer cases are due to environmental, dietary, or lifestyle factors. Accordingly, these cases maybe avoided by appropriate modifications. In addition, active chemoprevention has become a major interventional approach following the epidemiological observation of a beneficial effect of nonsteroidal anti-inflammatory drugs (NSAIDs) in colon cancer prevention. This is chiefly due to the inhibition of the cyclooxygenase (COX) enzymes. The COX enzymatic system includes two isoenzymes, COX-1 and COX-2, that convert arachidonic acid to prostaglandins. COX-1 is constitutively expressed and synthesizes cytoprotective prostaglandins in the gastrointestinal tract. COX-2 is inducible by the oncogenes ras and scr and other cytokines; it is overexpressed in human cancer cells in which it stimulates cellular division and angiogenesis and inhibits apoptosis. NSAIDs restore apoptosis and decrease tumor mitogenesis and angiogenesis. Most cancer cells have been found to exhibit overexpression of COX-2. Epidemiological studies showed a lower risk of developing cancer of the colon, breast, esophagus, and stomach following the ingestion of NSAIDs. The use of NSAIDs in low dose was associated with a statistically significant decrease in the risk of adenomatous polyps and of overt colon cancer. The regressive effects of sulindac on foci of aberrant crypts in the colon (considered to be precursors of adenoma), and on adenocarcinoma of the colon, are of particular interest because this NSAID does not have an inhibitory effect on COX. This may support the view that the antineoplastic effect of NSAIDs may also be due to a mechanism other than COX-2 inhibition. In breast cancer, large cohort studies reported a 40 to 50% reduced risk of developing cancer, a smaller size of the primary tumor, and a reduction in the number of involved axillary lymph nodes. Similar findings have been reported in the esophagus and stomach, but not in gastric cardia adenocarcinoma. The recent development of selective COX-2 inhibitors resulted in better clinical tolerance than that associated with NSAIDs in general, with the absence of gastrointestinal side effects known to occur after the inhibition of COX-1. Encouraging results have been obtained with these new agents in familial adenomatous polyposis, colon, breast, and prostate cancer. PMID- 12086407 TI - Prostaglandins and the regulation of tumor growth. AB - Increased expression of inducible cyclooxygenase (COX-2) is associated with a wide variety of tumors. In addition, inhibitors of COX have shown a great deal of promise in vitro and in animal models as potential antitumor therapies. COX enzymes use the substrate arachidonic acid to produce prostaglandin (PG)H2, the precursor to all the prostanoids. Therefore, the release of individual prostanoids depends on the abundance and functional coupling to individual PG synthase isoenzymes. Colony stimulating factors (CSFs) are also potential antitumor agents via their ability to augment the immune response. When COX-2 is expressed, the CSF, granulocyte macrophage (GM)-CSF, and granulocyte (G)-CSF are exquisitely sensitive to endogenous PGs. In addition, the ability of COX-2 to suppress GM-CSF release is mediated via traditional IP/EP prostanoid receptors linked to cAMP-dependent pathways. Therefore, inhibition of COX-2 in tumors may have the important side effect of enhancing the immune response. Recently, novel signaling pathways for PG derivatives have been discovered; in particular the PGD2 dehydration product 15-deoxy-delta(12,14)-(15d)-PGJ2 was identified as a ligand for the nuclear receptor/transcription factor, peroxisome proliferator activated receptor (PPAR)-gamma. PPARgamma is present at high levels in a number of tumors, and is also present in endothelial cells. 15d-PGJ2 as well as other nonprostanoid PPARgamma ligands are antitumor, and antiangiogenic, by dramatically inhibiting the growth of tumor cells and endothelial cells by either causing terminal differentiation, and/or by inducing apoptosis. We have recently found that, in addition to IP and EP ligands generated by COX-2, PPARgamma ligands similarly inhibit GM-CSF release. Effecting individual prostanoid pathways at the level of COX expression, profile of PG products produced or selective PG receptor activation may produce novel therapies, either dependent or independent of CSF release, to target cancers. PMID- 12086409 TI - Isolator Rodent Caging Systems (State of the Art): A Critical View. PMID- 12086408 TI - Isolator Caging Systems: Ventilated or Static? PMID- 12086410 TI - Mice Fed Radiolabeled Protein by Gavage Show Sporadic Passage of Large Quantities of Intact Material Into the Blood, an Artifact not Associated With Voluntary Feeding. AB - Bovine 2-microglobulin (Bov 2-M) is a small acid- and protease-resistant protein found in cows' milk that has the capacity to interact specifically with molecules of the mouse immune system. We conducted a series of investigations designed to determine whether a portion of orally ingested Bov 2-M could pass intact into the mouse circulatory system. Initial experiments feeding 125I-labeled protein by gavage to nonfed mice indicated that intact Bov 2-M appeared rapidly and at high amounts in the blood (as detected by use of direct sodium dodecyl polyacrylamide gel electrophoresis analysis of a 1-ml sample of plasma and phosphorimaging), but this result was observed sporadically and only in about a third of all animals, with no detectable protein appearing in the plasma of the remaining animals. Identical results were obtained when the mice were lightly anesthetized prior to gavage. When a larger protein, 125I-labeled bovine serum albumin (BSA), was fed by gavage in combination with the labeled Bov 2-M, surprisingly large fragments of the BSA (. 50 kDa) appeared in the plasma whenever the Bov 2-M appeared, again in about a third of the animals. We hypothesized that the passage of large quantities of Bov 2-M into the blood of some mice was due to an intermittent artifact of the gavage feeding (likely spillover into the lungs), and therefore, we developed an alternate approach based on voluntary feeding. Mice were easily taught to drink small samples of liquid voluntarily by removing water for 12 h, then manually generating droplets at the end of a 1-ml syringe. When a mixture containing 125I-labeled Bov 2-M and BSA was fed in this manner, labeled protein could not be detected in any animals by direct analysis of a 1-ml sample of plasma. However, when immunoprecipitation was carried out on a larger volume of plasma (100 ml), intact Bov 2-M could be found in the blood of all animals, whereas similar immunoprecipitation and analysis for BSA did not yield any signal. We conclude that the common method of gavage feeding mice to assess absorption of orally ingested material can lead to artifacts not seen when the same agent is consumed under more natural circumstances. These observations may have relevance for experiments where oral tolerance is assumed to be induced in rodents by repeated gavage feeding, whereas the same effects cannot be obtained in humans by voluntary feeding. PMID- 12086411 TI - Effective Strategy for Evaluating Tactile Enrichment Devices for Singly Caged Macaques. AB - Since the mandate for providing environmental enrichment for nonhuman primates was included in the Animal Welfare Act, numerous articles and suggestions have been put forth covering tactile devices and creative cage arrangements. For larger primate facilities and research programs environmental enrichment evaluation is usually accomplished by enrichment technicians or behaviorists. However, for the smaller facilities or programs, the ability to formulate and document an enrichment program can be very difficult due to budget or personnel constraints. We present a simple, yet effective, tactile device scoring system used with singly caged macaques indicating that creating and documenting enrichment ideas can be accomplished without a large personnel and budgetary commitment. We believe this strategy will help programs meet the regulatory requirements with relative ease. PMID- 12086412 TI - Puzzle Feeders and Gum Feeders as Environmental Enrichment for Common Marmosets. AB - Common marmosets (Callithrix jacchus jacchus) are highly social New World monkeys that consume a principally gummivorous and insectivorous diet. We examined the efficacy of two types of foraging devices, Puzzle-Feeders(tm) and gum feeders, as environmental enrichment for marmosets housed singly (n = 16) or in sibling (n = 4) and heterosexual (n = 8) pairs. In experiment 1, marmosets were exposed to each of the two types of foraging devices for three hours, once per week for two weeks. Thirty-minute observations were conducted at the beginning and end of each exposure period. Marmosets in all housing conditions experienced significant reductions in the frequency of stereotyped pacing and significantly less time sitting still while exposed to the foraging devices. Marmosets experienced significantly lower levels of feeder use and significantly more time sitting still at the end of the three-hour exposure than at the beginning. Marmosets that were singly or sibling housed used the devices the most and had the largest reductions in time spent sitting still during enrichment. In experiment 2, singly housed marmosets were given two types of gum feeders, a wooden and a Gumabone(tm) gum feeder, each for a week-long period. Thirty-minute observations were conducted three times per week immediately after loading the feeders with fresh gum. The wooden gum feeders were heavily gouged during the week-long exposure, although significantly less use of both types of gum feeders was observed on the third and fifth days. These results indicated that marmosets in variable social housing conditions can benefit from environmental enrichment additional to social housing, and that foraging enrichment promotes increased non-stereotyped movement and decreased pacing in this species. PMID- 12086413 TI - Biomethod for Obtaining Gastric Juice and Serum From the Unanesthetized Guinea Pig (Cavia porcellus). AB - Blood sample collection and gastric intubation of guinea pigs have been considered difficult techniques that require anesthesia. We developed methods to sample blood and gastric juice from manually restrained, unanesthetized guinea pigs. To collect gastric juice, the guinea pig is restrained in vertical position by an assistant, who keeps the guinea pig's head in extreme dorsoflexion by use of a strip of gauze looped around the top incisors. The operator uses a second strip of gauze to control the lower jaw, and inserts a 5- or 6-F infant feeding tube down the throat, being careful not to deviate to either side of the buccal cavity. The tube slides directly down the esophagus of a correctly positioned guinea pig, and up to 5 ml of gastric juice can be withdrawn, using a syringe attached to the feeding tube. Blood sample collection was done by jugular venipuncture of manually restrained mesmerized guinea pigs; up to 2.5 ml of blood can be collected via this route. We have used these techniques on more than 50 guinea pigs ranging from 2 weeks to 18 months of age, and obtained weekly blood and gastric juice samples with no resultant morbidity or mortality and minimal distress. Repeated gastric juice and blood collections can be made safely from manually restrained, unanesthetized guinea pigs. PMID- 12086414 TI - Influence of Age on the Electrocardiographic Waves in Taiwanese Lan-Yu Miniature Pigs. AB - The purpose of the study reported here was to determine baseline information of the electrocardiogram (ECG) and the influence of age on cardiac function in Taiwanese Lan-Yu (TLY) miniature pigs. Non-anesthetized TLY minipigs (from birth to 6 months of age) were placed on a webbed stanchion, simulating a standing position, for acquisition of an ECG, using six limb leads (I, II, III, aVR, aVL, and aVF), with each as a single-channel ECG recorder. The P and T waves obtained from leads I, III, and aVL were useful in determining the subtle cardiac changes during maturation of TLY minipigs. Interestingly, changes in the QT interval analyzed from all 6 leads were almost indistinguishable. Shortening of the QT interval was induced (p < 0.05) between days 1 and 7 of postnatal life. The QT interval lengthened to a steady state at day 60, and paralleled pigs' physical maturation. The longer QT interval was conversely correlated to heart rate as pigs matured. In the QRS complex interval, only lead aVR was significantly decreased at day 7 (p < 0.05). Further changes in the QRS interval from day 21 were not observed in any lead. Because the duration of the ventricular complex represents the period required for the excitation front to reach the terminals of the Purkinje fibers in the ventricular myocardium, the increase in QRS interval observed within 21 days of birth could be attributed to an increase in the thickness of ventricular myocardium. The data suggest that cardiac maturation was achieved at 60 days of age, although the body weight of minipigs continued to increase beyond 60 days of age. Because the body weight of these newly developed TLY minipigs can be maintained within 25 to 30 kg at one year of age and their major ECG findings did not significantly differ from those of domestic pigs and humans, they may be useful as a model for cardiovascular and pharmacologic research. The similarity of ECG profiles between pigs and humans also was evaluated. PMID- 12086415 TI - Outer Cortical Necrosis Associated With Renal Artery Constriction in the Kidney of a Dog. PMID- 12086416 TI - Oropharyngeal Necrobacillosis with Septic Thrombophlebitis and Pulmonary Embolic Abscesses: Lemierre's Syndrome in a New Zealand White Rabbit. AB - Lemierre's syndrome, oropharyngeal infections induced by anaerobic bacteria, leading to fatal septic thrombophlebitis of the internal jugular vein and pulmonary embolic abscesses in humans, was diagnosed in a 6-month-old, male, New Zealand White rabbit. After acute onset of anorexia, lethargy, and depression, the rabbit died suddenly despite emergency clinical care. Necropsy revealed swelling, necrosis, and abscess in the soft tissues around the left caudal mandibular ramus, oral mucosa, and molar teeth, with systemic embolic abscesses and necrosis, especially in the jugular vein, lungs, and brain. Histologic examination revealed necrosis and embolic abscesses with filamentous bacteria in the mandibular soft tissues, salivary gland, jugular veins, alveolar bone and marrow, periodontal tissues and dental pulp, oral and pharyngeal mucosa, lungs, brain, liver, myocardium, meninges, and small intestine. Bacterial culture of the mandibular abscess and heart blood yielded Fusobacterium necrophorum. PMID- 12086418 TI - Gastric Perforation in WAG/Rij Rats after Esophagojejunostomy. PMID- 12086417 TI - Epidermotropic Lymphoma (Mycosis Fungoides) in an ICR Mouse. AB - A case of epidermotropic lymphoma with systemic spread into lymph nodes and visceral organs was observed in a 7- to 8-month-old, female ICR mouse. The mouse developed progressive and generalized alopecia and lymphadenopathy of several weeks' duration. The affected skin was markedly and diffusely thickened, with multiple serous to hemorrhagic crusts, ulcerated plaques, and raised nodules. Microscopically, random and/or perivascular infiltration of pleomorphic lymphoid cells was present in the skin, lymph nodes, thymus, spleen, lungs, and liver. On cytologic examination, the lymphoid cells were similar in all affected tissues, and had hyperchromatic and irregularly oval, cleaved, and occasionally convoluted nuclei, approximately 6 to 9 mm in diameter. On immunohistochemical examination, most infiltrating cells were labeled with anti-CD3 (panT cell) antibody. A smaller proportion of the cells (, 5%) were labeled with anti-CD79a (panB cell) antibody. PMID- 12086419 TI - Hemagglutination Inhibition (HAI) Assay. PMID- 12086420 TI - Special Feeding and Care of Senescent Spontaneously Hypertensive Rats. AB - This study investigated the impact of feeding methods on body weight of senescent female spontaneously hypertensive rats (SHRs) and showed that supplementing powdered feed was useful as they approached heart failure at 22 to 23 months of age. SHRs are genetically predisposed to systemic hypertension and will, with age, progress into complete heart failure resulting in death. Close to the time of heart failure, some rats experienced a loss of appetite and weight loss. It was postulated that either elevated blood pressure, age-associated health issues, or the effort required to access pelleted food prevented the rats from eating properly, resulting in malnutrition and weight loss. As they aged, the rats benefited from the addition of powdered food to ensure that body weights remained stable and to prevent malnutrition that could lead to premature death. Animals were fed commercially available rat chow pellets until they showed persistent signs of weight loss or a lack of interest in their food. At that time, the rats were also given powdered rat chow in shallow bowls to facilitate the eating and the digestion of their food. The rats were weighed weekly to confirm they consumed sufficient calories daily and to ensure that the change to the powdered chow was having the desired effect. Prior to being fed the powdered rat chow, the rats had shown signs of progressive weight loss. After starting the powdered chow, the rats either maintained or gained weight. This study shows that as the female SHR matures, special care and handling is key to maintaining body weights and good health. With only modest changes in routine (i.e., powdered food) and an attentive eye on the rats' daily activities, it was possible to maintain these senescent female SHRs in a healthy condition until the termination of the study or onset of heart failure. PMID- 12086421 TI - The Use of a Percutaneous Endoscopic Gastrotomy (PEG) Tube To Reverse Fatal Fasting Syndrome in a Cynomolgus Macaque (Macaca fascicularis). AB - An aged, overweight, female cynomolgus macaque (Macaca fascicularis) was diagnosed preliminarily with fatal fasting syndrome based on a history of chronic anorexia, weight loss, hepatomegaly, elevated serum alkaline phosphatase, bilirubinuria, and diffuse increased hepatic echogenicity. Histopathologic examination confirmed the diagnosis of severe hepatic lipidosis. A percutaneous endoscopic gastrotomy (PEG) tube was inserted after a month of orogastric tube feedings failed to stimulate normal appetite. Within 2 weeks of twice daily PEG tube feedings, the animal began to eat voluntarily, and PEG tube feedings were discontinued. The PEG tube was removed without incident. Periodic percutaneous ultrasound-guided hepatic biopsies revealed partial resolution of the hepatic lipidosis during the following 18 months. PEG tube placement should be considered as an alternative treatment to provide nutritional support in macaques with fatal fasting syndrome. PMID- 12086422 TI - A Modified Cage to Minimize Catheter Contamination in the Chronically Catheterized Baboon. AB - Here we describe modifications made to an 8 sq. ft. aluminum baboon cage to allow removal of a chronically cannulated baboon from the cage without disconnecting the catheter connections. The novel system minimizes potential contamination of the intravenous catheters in an immunosuppressed baboon model and permits removal of the animal for cage changes and transport to a distant facility for experimental manipulation. PMID- 12086423 TI - Progressive Anemia in a Pig-tail Macaque With AIDS. AB - The present report is a diagnostic exercise of a case of progressive anemia in a pigtail macaque with AIDS. In summary, the immune suppression of the macaque by the KUSHIV-1B virus prevented an effective immune response to simian parvovirus (SPV), resulting in persistent infection with progressive anemia. Simian parvovirus should be considered in the differential diagnosis of anemia in macaques typically used in research. PMID- 12086424 TI - Indirect Fluorescent Antibody (IFA) Assay. PMID- 12086425 TI - The Effects of Commercially Available Sodium Pentobarbital Formulations on Plasma Hemoglobin Levels in New Zealand White Rabbits. AB - Sodium pentobarbital (Abbott Laboratories) was widely used as an intravenous (IV) anesthetic in rabbits, but it was removed from the market in 1995. Many commercially available alternatives, including Nembutal(r) (Abbott Laboratories), Euthanasia-5(r) (Veterinary Laboratories, Inc.), and Euthanasia-6(r) (Veterinary Laboratories, Inc.), contain sodium pentobarbital. However, all of these products produced an unacceptable level of hemolysis in antisera collected from anesthetized rabbits. The experimental design of this study included evaluating each component of these pentobarbital formulations to determine the exact cause of hemolysis. The goal of this study was to find an alternative formulation of sodium pentobarbital that did not produce hemolysis in rabbit sera obtained from terminal bleeds. Specific ingredients in all formulations included sterile water, alcohol, propylene glycol (PG), and sodium pentobarbital in various combinations and concentrations. Study animals were inoculated intravenously with either sterile water, 10% ethanol, 40% ethanol, 2.5% PG, 40% PG, or isotonic saline as a control. PG at 40% concentration produced the most severe hemolysis, followed by 40% ethanol and sterile water. Levels of hemolysis caused by 10% ethanol and 2.5% PG did not differ statistically from those produced by isotonic saline. Subsequently, our laboratory adopted a pentobarbital formulation consisting of sodium pentobarbital dissolved at a concentration of 50 mg/ml in 10% ethanol and sterile water. This formulation is non-hemolytic in rabbits. PMID- 12086426 TI - EEG Recordings in Anesthetized Rabbits: Comparison of Ketamine-Midazolam and Telazol With or Without Xylazine. AB - Electroencephalographic (EEG) recordings were performed on five New Zealand White rabbits anesthetized by using four intramuscular drug combinations: 1) ketamine (30 mg/kg) and midazolam (0.2 mg/kg), 2) ketamine (30 mg/kg), midazolam (0.2 mg/kg), and xylazine (3 mg/kg), 3) Telazol (20 mg/kg), and 4) Telazol (20 mg/kg) and xylazine (3 mg/kg). All four combinations were administered randomly to each rabbit. To evaluate anesthesia depth, response to toe pinch and various measurements from the recordings were assessed before and after injection. For each EEG recorded epoch, b/d ratios and the spectral edge frequencies (SEF) at 80% and 95% were measured. Results show that after injection of combinations without xylazine, b/d ratios and SEFs decreased only slightly; concurrently the withdrawal reflex remained present. Adding xylazine decreased the b/d ratios (p, 0.001) and the SEFs at 80% (p, 0.001) and 95% (p, 0.001). No withdrawal reflex was observed for 30 min after injection of ketamine-midazolam-xylazine and for 60 min after administration of Telazol-xylazine. Therefore, EEGs may be used to evaluate depth of anesthesia when using injectable drug combinations in rabbits.Abstract> PMID- 12086427 TI - Cushing's Syndrome with Concurrent Diabetes Mellitus in a Rhesus Monkey. AB - Cushing's syndrome is the clinical expression of the overproduction of glucocorticoids and is well recognized in both human and veterinary medicine. Spontaneous diabetes mellitus is well known in Macaca spp., however the occurrence of hyperadrenocorticism and diabetes mellitus concurrently in macaques has not been reported previously. This unusual case presents a rare opportunity to examine the relationships between two important endocrine diseases in a nonhuman primate. A male, 14-year-old rhesus macaque (Macaca mulatta) was diagnosed with hyperadrenocorticism and concurrent diabetes mellitus. Initially, the monkey had mildly elevated blood glucose values upon routine semi-annual physical examination. Further diagnostic work-up demonstrated hypercortisolism. Adrenocorticotropic hormone-dependent Cushing's syndrome was subsequently diagnosed in light of results from dexamethasone testing, magnetic resonance imaging, and computed tomography scans. A therapeutic course of L-deprenyl (Anipryl(r)) was begun, and 8 weeks later, insulin therapy was initiated. The patient responded well to insulin therapy, however the dosage was rapidly increased. After 6 months, Anipryl(r) therapy was determined to be of little or no value, and ketoconazole was selected as the drug of choice to control the hypercortisolism. The monkey has shown remarkable improvement with the dual therapies of insulin and ketoconazole. Approximately 2 months after the initiation of ketoconazole therapy, the animal was returned to an experimental protocol under the conditions of twice-daily treatment and strict dietary control. The ongoing plan for clinical management includes periodic blood glucose and liver function surveillance. PMID- 12086428 TI - Factors Optimizing the Use of Subcutaneous Vascular Access Ports in Weaned Pigs. AB - Young, recently weaned pigs were implanted with subcutaneous vascular access ports in experiments designed to determine the relative bioavailability of lead from contaminated soils. In nine experiments, a total of 72 subcutaneous vascular access ports were implanted. Complications were observed in 38.9% of the implanted pigs. Changes in the protocol were implemented in an effort to reduce the rate of complications. The majority of complications were septic processes, including localized infections around the site of the catheter and injection port, hematogenous pulmonary abscessation, endocarditis, and septic arthritis. Staphylococcus aureus, S. hyicus, b-hemolytic Streptococcus species, and Actinomyces pyogenes were cultured from septic processes in these pigs. Our findings suggest that the health of the skin at the time of surgery, the experience of the surgeon, and the immunologic status of the animal influence the risk of septic complications in pigs implanted with subcutaneous vascular access ports. PMID- 12086429 TI - A Nest Box to Facilitate Excreta Collection From Mouse Dams Through Pregnancy, Parturition, and Lactation. AB - Standard metabolism cages are inadequate for collecting excreta from dams during parturition because newborn pups can fall through the grating into the excreta collection area and out of reach of the dam. A nest box was designed that facilitates excreta collection from mouse dams continuously housed in metabolism cages from conception, through parturition, and into lactation and provides a safe, warm environment for pups during their first week of life. The nest box was tested by using pregnant and lactating mice of two varieties of strain 129/SvJ, metallothionein-normal and metallothionein-knockout; non-pregnant mice were used as controls. Pregnant mice (with nest box) and non-pregnant mice (without nest box) each twice received a solution of 109CdCl2 by gavage. Dams with nest boxes fastidiously urinated and defecated outside the nest box. The percentage of gavage 109Cd dose recovered in dam feces was the same after the first gavage (mean6SE, with nest box through parturition, 95%66%; n=5) as after the second gavage (mean6SE, without nest box, 95%66%; n=5). Weights and percentage weight gain of mouse dams were independent of housing conditions (metabolic cage with next box vs. conventional polycarbonate caging). Furthermore, pup growth and survival were unaffected by the inclusion of the nest box or by its removal at 1 week after birth. Therefore, the described nest box provides for the first time a way to quantitatively collect excreta from mouse dams through pregnancy, parturition, and the early postnatal period. Additional experiments are needed to test its application to other animal species and strains of mice, including those with poor mothering behavior. PMID- 12086430 TI - Urinary Bladder Catheterization of Female Mice and Rats. AB - Catheterization of the urinary bladder of rats and mice is necessary for a variety of clinical and experimental reasons. The anatomy of the female mouse and rat is unique in that the urinary orifice is external and just anterior to the vaginal opening. This characteristic makes the bladder of these species easier to catheterize than that of other species. Methods cited in the literature do not adequately describe techniques that facilitate ease of urinary bladder catheterization (1). We describe a simple technique for urinary catheterization in adult female mice and rats. PMID- 12086431 TI - High Mortality in Zebrafish (Danio rerio). AB - A group of 100 adult zebrafish were housed in a new system at a stocking density of 20 fish per tank. Four weeks after arrival, 15 fish presented with petechial hemorrhages and ulceration on the surfaces of the skin. Samples of the fish were collected for histopathology, fungal culture, and bacterial culture and sensitivity. Water samples were analyzed for pH, ammonia, nitrite, and submitted for bacterial and fungal culture. Histologically, the epidermis had multiple areas of ulceration and mononuclear cell infiltrate. Gram-positive bacteria were observed beneath the surface of the skin and surrounding the outer aspect of the spinal cord. Both Aeromonas hydrophila and A. sobria were isolated from the affected fish, and a diagnosis of motile aeromonad septicemia (MAS) was made. Water from the tanks had a nitrite level of 1-5 ppm, a toxic concentration that indicated poor water quality. Because the housing system had been seeded with Nitrobacter spp. and Nitrosomonas spp. only 2 weeks prior to the arrival of the fish, a lack of colonizing nitrifying bacteria was deemed to be the cause of the high nitrite level, which, along with over-crowding, stressed the fish and increased their susceptibility to MAS. No further cases of septicemia were observed once the nitrite level and stocking density were reduced. PMID- 12086432 TI - Enzyme-Linked Immunosorbent Assay (ELISA). PMID- 12086433 TI - Assessing the Value of Television as Environmental Enrichment for Individually Housed Rhesus Monkeys: A Behavioral Economic Approach. AB - The goal of this study was to evaluate television as a source of environmental enrichment for individually housed rhesus monkeys (Macaca mulatta) by using the concepts of behavioral economics. Phase I entailed the use of operant conditioning to assess the behavior of eight rhesus monkeys given the opportunity to control their environment through lever activation of a television (TV). Success in shaping was variable, and only two animals successfully acquired lever pressing. Phase II used an alternating reinforcement/ extinction procedure as a control method to determine the degree to which lever pressing depended on TV presentation. Both animals responded with more lever pressing on the days when lever pressing produced TV. The first animal, tested with the alternating reinforcement/extinction procedure for 12 weeks yielded a mean significant difference of 3.85 (p = 0.036); the second assessed for 9 weeks was associated with a mean significant difference of 6.0 (p = 0.018). Therefore, TV (and not lever pressing itself) was positively reinforcing. The final phase of the study progressively increased the fixed ratio (FR) from 1 to 8. Linear regression of the data points, plotted as the log of price (or FR) vs the consumption of TV, revealed a significantly negative slope (-2.179, p, 0.05) and accounted for 89% of the variance. The negative demand curve suggested that TV is not a valued commodity and is highly elastic. TV provided to individually housed rhesus monkeys appears to be a weakly positive reinforcer for some animals, which may contribute to overall environmental enrichment. PMID- 12086434 TI - Genetic Contamination of an SJL/J Mouse Colony: Rapid Detection by PCR-based Microsatellite Analysis. AB - Six simple sequence-length polymorphisms (SSLPs), or microsatellites, generated by polymerase chain reaction (PCR), were used to detect genetic contamination of an SJL/J mouse colony in a faster and more economical way than standard immunologic and biochemical markers. The marker set was chosen to assess SJL authenticity and to distinguish a possible outcross with BALB/c, the only other albino strain maintained at the facility. Evidence of genetic contamination as a result of an outcross with BALB/c was found, and the results were ready within a workday. PMID- 12086435 TI - Determination of Agreement Between Laboratory Instruments. AB - The present study was conducted to illustrate the utility of Bland-Altman plots for use by our laboratory staff and other non-statisticians in assessing the agreement between values measured by using two different laboratory instruments. A high degree of agreement reflects acceptable interchangeability of equipment and minimal effect on clinical decision-making. We have summarized literature that suggests that the regression line and correlation coefficient used with regression analysis, although commonly employed, are not appropriate first choices for evaluating agreement. Using the ABL 500 Radiometer and i-STAT Portable Chemistry Analyzer, we evaluated pH, PCO2, and TCO2. Bland-Altman plots were simple to produce, were not mathematics-intensive, and provided an easily interpreted, graphical answer to the question of agreement between instruments. For purposes of clinical decision-making, the ABL and i-STAT machines were found to be in good agreement for the tests evaluated. PMID- 12086436 TI - Comparison of Cardiovascular Parameters and/or Serum Chemistry and Hematology Profiles in Conscious and Anesthetized Rhesus Monkeys (Macaca mulatta). AB - The rhesus monkey is often used in pre-clinical research, and such studies frequently involve a variety of anesthetic conditions. Therefore, it is important to determine baseline physiologic blood chemistry and cardiovascular parameters in anesthetized animals to facilitate appropriate comparisons. The present study compares the cardiovascular parameters, hematology, serum chemistry, and blood gas levels of rhesus monkeys anesthetized with pentobarbital, isoflurane, ketamine, and propofol. Hematology, serum chemistry, and blood gas levels were unaffected by the four anesthetic regimens. However, because of its formulation, propofol is inappropriate for use in animals in which changes in tryglycerides will be evaluated. Compared to those in conscious, unrestrained monkeys, heart rates were higher in anesthetized animals, but the rates of anesthetized animals were similar regardless of anesthetic agent used. In contrast, mean arterial blood pressure was lower in animals anesthetized with pentobarbital, propofol, or isoflurane than in the conscious monkeys. However, mean arterial pressure of ketamine-anesthetized monkeys was similar to that of the conscious monkeys. PMID- 12086437 TI - A Nonsurgical Method for the Placement of Transpyloric (Nasoenteric) Feeding Tubes Using Fluoroscopic Guidance in Dogs. AB - Transpyloric tube feeding is a commonly used form of nutritional support when patients are unable to tolerate oral or gastric feeding. Although nasoenteric placement of transpyloric feeding tubes is usually accomplished in humans, anatomic variations in laboratory species have necessitated surgical laparotomy for transpyloric tube placement. We have developed a reliable fluoroscopic technique for the nonsurgical placement of transpyloric feeding tubes in dogs. This technique was developed in an experimental setting and currently is being used in the treatment of canine patients at the Auburn University College of Veterinary Medicine Small Animal Clinic. After nasal passage of a weighted 8 French feeding tube with stylet into the stomach, dogs are placed in sternal recumbency, and the tube is advanced toward right side of the fundus under fluoroscopic visualization. The animal is then placed in left lateral recumbency, and the stomach is insufflated with 20-30 ml/kg air through the tube. The pylorus is visualized as air passes from the stomach into the proximal small intestine, and the tube is advanced across the pylorus and into the proximal intestine. Placement has been successful in anesthetized and awake animals and in dogs with a variety of illnesses. Complications include bile leakage from the externalized opening of the tube, with subsequent cellulitis and retrograde movement of the tube when placement is proximal to the caudal duodenal flexure. This technique can be used in clinical veterinary medicine and nutritional research studies involving canine models and avoids abdominal surgery for the placement of intestinal feeding tubes. PMID- 12086439 TI - The Importance of Keeping the "Science" in the American Association for Laboratory Animal Science. PMID- 12086438 TI - Quaternary Ammonium-Induced Cutaneous and Gastrointestinal Mucosal Lesions in a Dog. AB - A dog developed severe ulcerative dermatitis and upper gastrointestinal mucosal ulceration after accidental exposure to a concentrated quaternary ammonium disinfectant solution. Secondary problems included dyspnea and cholestasis. Prolonged intensive care was required and included gastrostomy tube placement via endoscopy, extensive wound care, and systemic antibiotics. In contrast to earlier reports and although the offending compound was in higher concentration, contact irritation was much more severe yet no overt systemic toxic effects were noted. This case illustrates the great caution that must be exercised when handling commonly employed laboratory germicides. PMID- 12086440 TI - The Genetic Engineering Revolution. PMID- 12086441 TI - Demodex musculi in the Skin of Transgenic Mice. AB - Although infestations by a number of Demodex mite species have been described in mice, the occurrence of Demodex musculi infestation was last reported by Hirst in 1917. This communication describes the occurrence of D. musculi infestation in two lines of transgenic mice and their F1-hybrid offspring. We first found the Demodex mite in mouse hair samples collected during efficacy screenings in an ongoing ectoparasite treatment trial for the fur mite Radfordia affinis. An investigation was undertaken to determine the extent of the Demodex infestation within the facility and the original source of the parasite. D. musculi was found in three of the four mouse genotypes present in the index room and in one of these genotypes in two other rooms. The mite was not found in sentinel mice, other strains, or stocks within the facility. The mites were more easily recovered from the immunodeficient B6,CBA-TgN(CD3E)26Cpt transgenic (Tg) and the hybrid double-Tg (B6,CBA-TgN(CD3E)26Cpt x B6,SENCARB-TgN(pk5prad1)7111Sprd)F1 mice than from the B6,SENCARB-TgN(pk5prad1)7111Sprd Tg mouse, which is believed to be immunocompetent despite its thymic abnormalities. Histopathologic examination showed D. musculi superficially in hair follicles but not in the preputial or clitoral gland or in serial sections of the head, eyelids, or ears, the locations favored by other mouse demodicids. Physical and microscopic examination revealed no dermatitis. The immune deficiency in the B6,CBA TgN(CD3E)26Cpt mouse probably provided the permissive host conditions that contributed to the proliferation and subsequent detection of the Demodex. Preliminary transmission experiments conducted with other immunologic mutant mice and our sentinel strain demonstrated variation in mite transfer and in either detection or maintenance of infestation when na ve mice were housed with those carrying D. musculi. The original source of D. musculi was not conclusively identified, but this parasite appears to be of low pathogenicity in the examined genotypes. PMID- 12086443 TI - Influence of Blood Collection Sites on Plasma Glucose and Insulin Concentration in Conscious C57BL/6 Mice. AB - Blood collection from conscious, unanesthetized mice is often performed during the drug development process. The site of collection may influence the parameter(s) of interest. To investigate the potential influence of collection site on plasma glucose and insulin, a study was conducted to compare plasma glucose and insulin concentrations in blood samples collected without anesthesia from the retroorbital sinus versus the tail vein in 10- to 12-week-old female C57BL/6 mice. Two experiments were performed. In the first experiment, mice were randomized to be bled from the tail vein then the retroorbital sinus or vice versa in a balanced two-period crossover design. In this experiment, the retroorbital and tail vein bleeds were performed a few minutes apart. The second experiment was similar to the first, except the bleeds were performed 1 week apart. Overall, retroorbital collection yielded lower glucose levels (p, 0.001) and higher insulin levels (p, 0.001) than did tail vein collection. The minimum difference in measured glucose in a retroorbital collection versus a tail vein collection, after adjusting for the effect of sequential bleeding was -97 mg/dl; the maximum was 98 mg/dl. We estimate that about 98% of observations taken under similar conditions would fall in this interval. The minimum difference in measured insulin in a retroorbital collection versus a tail vein collection was 0.6 ng/ml; the maximum was 7.3 ng/ml. We estimate that about 98% of observations taken under similar conditions would fall in this interval. PMID- 12086442 TI - The Influence of Euthanasia Methods on Rat Liver Metabolism. AB - An important consideration in any terminal experiment is the method for euthanizing animals. Although the prime consideration is that the method is humane, some methods can have a dramatic impact on experimental outcomes. To determine the optimal method of euthanasia in metabolic experiments, a physical method (decapitation), two asphyxiation methods (CO2 and O2/CO2), and anesthetic (isoflurane) exposure followed by exsanguination were compared for their effects on liver metabolism. Changes in metabolism were monitored by following the activities of several key metabolic enzymes that are known to be susceptible to alterations in extracellular hormones as well as to changes in intracellular energy availability. The substrates and products of these enzymes also were monitored to better estimate their in vivo activity. Decapitated animals were used as the baseline for all comparisons. The results showed that euthanasia after exposure to 3 min isoflurane, euthanasia by exposure to a pure CO2 atmosphere for 2.5 min (CO2), and euthanasia by exposure to 1 min pure O2 followed by 2.5 min CO2 (O2/CO2) stimulated the enzymes involved in glycogen breakdown and glucose utilization. After CO2 or O2/CO2 asphyxiation, liver glycogen stores fell to approximately one-half those in the decapitated animals. No significant losses in liver glycogen were apparent after exsanguination under isoflurane anesthesia. In addition, differences between euthanasia methods were noted when the pattern of enzyme activity was compared: enzymes at the start of the glycolytic pathway were stimulated after CO2 or O2/CO2 euthanasia, but the terminal glycolytic enzyme was stimulated only after O2/CO2 euthanasia. Euthanasia by CO2 or O2/CO2 methods significantly decreased the regulatory enzyme of branched-chained amino acid degradation. This study clearly indicates that the method of euthanasia can have a dramatic impact on experimental data and, in particular, on liver metabolism. PMID- 12086444 TI - A Low-Cost Computerized System to Monitor Running Performance and Circadian Rhythms of Twenty Mice Simultaneously. AB - This paper describes the design and functioning of a low-cost computerized system for monitoring the voluntary activity of mice in running wheels. The required software is written in Turbo Pascal(r) and provided via the Internet (http://www.eur.nl/fgg/ch1/rodent.html). The system accommodates the simultaneous monitoring of 20 animals over a virtually unlimited period. Two applications of the system are presented; one monitors the circadian rhythm of mice, and the other tests muscle strength and endurance. PMID- 12086445 TI - Direct Laryngoscopy in Mice. AB - This report describes a noninvasive method for direct laryngoscopy in mice as small as 20 g by using commercially available equipment. A pediatric otoscope is inserted into the mouth of an anesthetized mouse to provide a clear view of the vocal cords. The procedure takes less than a minute and has been successful without complications in the 65 mice studied to date. The technique can be used to instill test substances into the lungs or to perform oral endotracheal intubation for resuscitation, airway support, or controlled ventilation. PMID- 12086446 TI - An Atypical Case of Mycobacterium bovis in a Cynomolgus Macaque (Macaca fascicularis) Imported From the Philippines. AB - An incidental case of Mycobacterium bovis was confirmed in a cynomolgus macaque from a shipment of 100 animals imported to the University of Southwestern Louisiana New Iberia Research Center from the Philippines. The macaque was euthanized 4 weeks into the quarantine period (October 1997) for failure to thrive and suspected melioidosis. Approximately 6 months later, on 9 March 1998, culture and antigenic probes from kidney tissues identified the etiologic agent as M. bovis. Five remaining cohort animals were euthanized after we obtained the results from the index case. Kidney tissues from two of the cohort animals were positive for M. avium. All animals had negative intradermal skin tests prior to euthanasia. All three animals positive for Mycobacterium were sero-negative for retroviruses. The unusual presentation of this case, coupled with the inability to detect disease by standard means, serves to emphasize the importance of follow up examination and culture of tissues obtained from imported non-human primates. PMID- 12086447 TI - Pyometra in a Siberian Polecat (Mustela eversmanni). AB - A 2-year-old Siberian polecat (Mustela eversmanni) from a breeding colony presented for ultrasound evaluation for pregnancy. It was paired with a male for 2.75 months and had remained absent of pregnancy signs when it was anesthetized and clinically evaluated. Until this time, the animal had eaten well and shown no outward signs of debility. On palpation, the animal had a fluid-filled tubular structure in the caudal abdomen, consistent in location and size with the uterus. No sign of vaginal discharge was present. Ultrasonography revealed 10 fluid filled evaginations (approximately 12 mm in diameter) of the uterine horns. A presumptive diagnosis of a fluid-filled reproductive tract and likely reproductive failure was made in light of the animal's history, its clinical signs, and the ultrasound findings. Euthanasia was performed because the animal was nonreproductive and might yield information relevant to the breeding colony as a whole. Necropsy of the polecat revealed a distended fluctuant uterus containing mildly odiferous, thick, yellow-green, purulent material. Histopathology confirmed the diagnosis of pyometra. A pure and heavy growth of Enterococcus fecalis was cultured from the uterine contents. In light of results from routine minimal inhibitory concentration antibiotic sensitivity screening, this isolate was resistant to all antibiotics tested in the standard teaching hospital screen. PMID- 12086448 TI - Rhabdomyosarcoma Associated With Incisor Malocclusion in a Laboratory Rat. PMID- 12086449 TI - Diarrhea in Neonatal Rats. AB - A litter of eight 10- to 12-day-old SPF Sprague-Dawley rat pups (Rattus norvegicus) was observed as runted, with rough, yellow-discolored hair coats and perineal scalding and soiling. The dam also had soft stool and perineal soiling. Both the dam and pups were active despite their unthrifty appearance. The clinical signs were consistent with diarrhea. Differential diagnoses for diarrhea in neonatal rats were considered, including bacterial, viral, and parasitic etiologies. The dam and pups were sacrificed, and the following diagnostic tests were performed: complete blood count, serology, necropsy, bacterial culture, and histopathology. Histopathology and culture results were consistent with a diagnosis of streptococcal enteropathy. PMID- 12086450 TI - Peritonitis in Nude Mice in a Xenograft Study. AB - After 30 Sim: (NCR) nu/nu mice were anesthetized with intraperitoneal tribromoethanol, they were injected with 106 human breast cancer cells or phosphate buffered saline (PBS) into the mammary fat pad through a surgical incision. Over the next six days, 24 of 30 mice became ill or died. Necropsies showed peritonitis, and a variety of bacteria were isolated from the peritoneal cavities of the mice. The tribromoethanol solution, PBS solution, and neoplastic cell line were all cultured and were microbiologically negative. Three immunocompetent mice were subsequently injected intraperitoneally with the same tribromoethanol solution used for the 30 nude mice. All three of the immunocompetent mice showed histologic evidence of peritonitis, but only one yielded positive peritoneal bacterial cultures and showed gross evidence of peritonitis. The tribromoethanol solution, previously reported to cause peritonitis, likely induced a chemical peritonitis in these mice, and the bacteria isolated were a secondary infection from normal gastrointestinal flora. We conclude that adequate mixing and storage of tribromoethanol are essential and that it is a safe and effective anesthetic when properly used. PMID- 12086451 TI - The Polymerase Chain Reaction (PCR). PMID- 12086452 TI - Effects of Crowding on the Social Behavior of Cuttlefish (Sepia officinalis). AB - To evaluate the effect of crowding on cuttlefish (Sepia officinalis), a benthic cephalopod, the behavior of captive-reared cuttlefish was monitored for a period of 1 month. One group of 6 cuttlefish was housed in a tank 6.1 m in diameter (4.87 m2 per cuttlefish); another group of 6 was housed in a tank 1.5 m in diameter (0.29 m2 per cuttlefish). Cuttlefish spaced themselves within each tank so as to avoid other cuttlefish. Those in the small tank hovered more and sat on the bottom less, showed more zebra patterns and Intense Zebra Displays (associated with aggression), ate less food, and were displaced by other cuttlefish 3 times as often as those in the large tank. Most aggression resulting in displacement was directed at females by males and sometimes resulted in physical injury to the females. Subjects' body patterns were highly predictable, using the following variables: activity, sex, tank, and number of nearby cuttlefish. Analysis of results indicated that behavior was strongly affected by housing conditions and suggested that this species is probably semi-solitary in natural conditions. PMID- 12086453 TI - Stereotactic Surgery and Long-Term Maintenance of Cranial Implants in Research Animals. AB - Most neuroscience research is performed by using anesthetized animals or tissue samples obtained from animals that have been euthanatized. However, study of many important issues requires the use of animals that are alert and capable of engaging in behavior. Various methods have been used to humanely perform neuroscience experiments that involved unanesthetized animals. These techniques often involve surgical implantation of an apparatus that permits direct manipulation of brain tissue or measurement of neurochemicals or neuronal activity in conscious animals. We describe here common surgical techniques used to prepare animals for long-term neuroscience studies, discuss several issues related to short- and long-term postoperative care of animals with implants, and offer suggestions that veterinary and research personnel can use to prevent or mitigate some common problems that may develop when preparing and maintaining animals for these studies. PMID- 12086454 TI - High Fungal Spore Load in Corncob Bedding Associated with Fungal-Induced Rhinitis in Two Rats. AB - Two types of commonly used contact bedding, aspen chip and corncob, were analyzed for the presence of fungal spores after two rats in a toxicology study were diagnosed with fungal rhinitis. A range of 700 to 5440 spores per g consisting of six species of fungus (Cladosporidium, Acremonium, Penicillium, Aspergillus, Fusarium, and Scolobasidium) were isolated from non-autoclaved corncob bedding obtained from two vendors. Negligible spore counts were detected in autoclaved corncob bedding and from either autoclaved or non-autoclaved aspen chip bedding. The present study demonstrates that high fungal spore counts may occur in untreated corncob bedding and that these may be associated with opportunistic disease in healthy, non-immunosuppressed animals. Autoclaving reduced the viable fungal spore load to an undetectable level. PMID- 12086455 TI - Environmental Conditions in a Chamber with Dry Ice. AB - Dry ice is used in research laboratories as a source of CO2 for euthanasia of rodents. The present study was performed to evaluate environmental conditions created in a standard (9-liter) bell jar. By using a portable gas analyzer and measuring at 7.5 cm above the platform, we evaluated the chamber filling rate at room temperature with a known quantity of dry ice, maintenance of 70% CO2, and concentration of CO2 maintained when the top of the chamber was removed and replaced. Ambient temperature in the chamber and temperature of the platform were also measured. Results indicated that 500 g dry ice was required to maintain the filling rate and CO2 concentration at levels recommended by the American Veterinary Medical Association Panel on Euthanasia; that after removal of an animal, the lid should be replaced for 1 min before placing another animal in the chamber; and that although ambient temperature in the chamber never fell below 178C during an 80-min period, the platform temperature decreased to 08C. If a chamber must be used as described above for more than 1 h, consideration should be given to alternating between two chambers and allowing a chamber to warm to room temperature before reuse. PMID- 12086456 TI - Reference Blood Chemistry Values Throughout the Lifespan of M. spretus. AB - In the present study, blood chemistry values for Mus spretus are reported and analyzed with respect to age and gender. Cholesterol values differ significantly (P = 0.01) between young (2 to 6 months) and aged (25 to 31 months) M. spretus, and direct or conjugated bilirubin values differ significantly (P = 0.01) between male and female animals. These findings indicate that cholesterol concentrations in M. spretus change with age and that direct bilirubin values differ depending on gender. PMID- 12086458 TI - Vaginal Cytology of the Lesser Bandicoot Rat (Bandicota savilei). AB - The lesser bandicoot rat (Bandicota savilei) is a wild field rat commonly found in Thailand. Vaginal smears were obtained from five colony-born animals twice daily for 28 days and stained with Giemsa stain. The external appearance of the vulva and appearance of the tip of vaginal swabs were recorded. All cells, including nucleated or keratinized epithelial cells and leukocytes, were quantified by percentage. No changes of the external genitalia were observed. The gross appearance of the vaginal swab may distinguish estrus and diestrus. Vaginal swabs contained a white or yellow material during estrus or diestrus, respectively. Changes in vaginal cytology at each stage of the estrous cycle of the lesser bandicoot rat were similar to those of the common laboratory rat. Estrus stages of the five rats occurred every 3 or 4 days. One female was introduced to a male after diestrus and produced a litter of six pups 27 days after pairing. PMID- 12086457 TI - Evaluation of Tiletamine-Zolazepam as an Anesthetic in Quail (Coturnix coturnix japonica). AB - The objective of the present study was to evaluate the effects of tiletamine zolazepam (TZ) administered alone or in combination with atropine, xylazine, and levomepromazine to quail (Coturnix coturnix japonica). The induction time, duration of hypnosis and anesthesia, and time to recovery were determined. The presence or absence of tremor, upper respiratory tract secretions, and excitability and the degree of muscular tone were also observed. The results showed that doses from 10 to 100 mg/kg TZ administered alone or in combination with xylazine or levomepromazine failed to produce anesthesia; only hypnosis was obtained in a dose-dependent manner. Immediately after injection of the drug, histopathologic examination of the site of drug injection indicated the presence of discrete acute focal myositis. After 21 days, a discrete fibrosis between muscle fibers was detected in the pectoral muscle as a sign of scarring. We conclude that the administration of TZ to a dose of 100 mg/kg does not produce anesthesia in quail. For noninvasive and minimally painful procedures requiring chemical restraint and recumbency, the recommended dose is 30 mg/kg. PMID- 12086460 TI - Improved Survival Rates in Mice that Received Prophylactic Fluids After Carcinogen Treatment. AB - During the development of a model for 7,12-dimethylbenz[a]anthracene (DMBA) induced mammary adenocarcinoma in mice, a high mortality rate was attributed to dehydration. Therefore, we compared the acute survival of mice given subcutaneous fluids prophylactically immediately following DMBA gavage to that of animals provided treatment only when clinical signs of dehydration were observed. Mortality in the prophylactically treated mice was 5% compared to 47% in animals treated only after the manifestation of dehydration. Prophylaxis with subcutaneous fluids significantly reduces mortality in DMBA-treated mice. PMID- 12086461 TI - Acute Paralysis Caused by Fibrocartilaginous Embolism in a Pigtail Macaque. AB - An adult female pigtail macaque was evaluated for acute flaccid rear limb paralysis with reduced patellar and anal reflexes. Despite supportive care, antibiotics, and corticosteroids, the monkey showed no improvement and was euthanized 7 days after the onset of clinical signs. Necropsy revealed a soft and swollen spinal cord within the L1 to L3 vertebrae. Microscopic examination of the spinal cord showed severe, diffuse ischemic necrosis, particularly of gray matter. Several small vessels within the spinal nerve root were partially to nearly totally occluded by material resembling fibrocartilage. To the best of our knowledge, this is the first reported case of fibrocartilaginous embolism in a nonhuman primate. PMID- 12086459 TI - Diagnostic Exercise: Fluffy, White, Cotton Candy-Like Growth on the Gills, Fins, and Skin of Salamanders (Ambystoma tigrinum). PMID- 12086462 TI - Spontaneous Renal Cell Carcinoma in a New Zealand White Rabbit. AB - The incidence of primary renal neoplasia in animals is quite low. Carcinomas are the most common primary renal tumors of dogs, cattle, and sheep. Among rabbit tumors, only uterine adenocarcinomas occur more frequently than do embryonal nephromas. However, spontaneous renal cell carcinomas in laboratory rabbits have only been reported once previously. We here report a second occurrence of a renal cell carcinoma in the laboratory rabbit. PMID- 12086463 TI - Combinatorial signaling by an unconventional Wg pathway and the Dpp pathway requires Nejire (CBP/p300) to regulate dpp expression in posterior tracheal branches. AB - The decapentaplegic (dpp) gene influences many developmental events in Drosophila melanogaster. We have been analyzing dpp expression in two groups of dorsal ectoderm cells at the posterior end of the embryo, in abdominal segment 8 and the telson. These dpp-expressing cells become tracheal cells in the posterior-most branches of the tracheal system (Dorsal Branch10, Spiracular Branch10, and the Posterior Spiracle). These branches are not identified by reagents typically used in analyses of tracheal development, suggesting that dpp expression confers a distinct identity upon posterior tracheal cells. We have determined that dpp posterior ectoderm expression begins during germ band extension and continues throughout development. We have isolated the sequences responsible for these aspects of dpp expression in a reporter gene. We have determined that an unconventional form of Wingless (Wg) signaling, Dpp signaling, and the transcriptional coactivator Nejire (CBP/p300) are required for the initiation and maintenance of dpp expression in the posterior-most branches of the tracheal system. Our data suggest a model for the integration of Wg and Dpp signals that may be applicable to branching morphogenesis in other developmental systems. PMID- 12086464 TI - FGF signaling regulates expression of Tbx2, Erm, Pea3, and Pax3 in the early nasal region. AB - Fgf8 is required for normal development of the nasal region. Here, we have used a candidate approach to identify genes that are induced in chick nasal mesenchyme in response to FGF signaling. Using an explant culture system, we show that expression of the transcription factors Tbx2, Erm, Pea3, and Pax3, but not Pax7, in nasal mesenchyme is regulated by ectodermal signals in a stage-dependent manner. Using beads soaked in recombinant FGF protein and an FGF receptor antagonist, we furthermore demonstrate that FGF signaling is necessary and sufficient for expression of Tbx2, Erm, Pea3, and Pax3, but has no effect on Pax7 expression. We also show that, within the nasal mesenchyme, competence to respond to FGF signaling is initially widespread and uniform but becomes restricted to regions normally exposed to FGF at later stages of development, coincident with changes in FGF receptor expression. Finally, we provide evidence that FGF8 also regulates Erm and Pea3 expression in the nasal placodes. Together, these results identify Tbx2, Erm, Pea3, and Pax3 as downstream targets of FGF signaling in the facial area and suggest that these genes may mediate some of the effects of FGF8 during development of the nasal region. PMID- 12086465 TI - Twist function is required for the morphogenesis of the cephalic neural tube and the differentiation of the cranial neural crest cells in the mouse embryo. AB - Loss of Twist function in the cranial mesenchyme of the mouse embryo causes failure of closure of the cephalic neural tube and malformation of the branchial arches. In the Twist(-/-) embryo, the expression of molecular markers that signify dorsal forebrain tissues is either absent or reduced, but those associated with ventral tissues display expanded domains of expression. Dorsoventral organization of the mid- and hindbrain and the anterior-posterior pattern of the neural tube are not affected. In the Twist(-/-) embryo, neural crest cells stray from the subectodermal migratory path and the late-migrating subpopulation invades the cell-free zone separating streams of cells going to the first and second branchial arches. Cell transplantation studies reveal that Twist activity is required in the cranial mesenchyme for directing the migration of the neural crest cells, as well as in the neural crest cells within the first branchial arch to achieve correct localization. Twist is also required for the proper differentiation of the first arch tissues into bone, muscle, and teeth. PMID- 12086466 TI - p107 and p130 Coordinately regulate proliferation, Cbfa1 expression, and hypertrophic differentiation during endochondral bone development. AB - During endochondral bone development, both the chondrogenic differentiation of mesenchyme and the hypertrophic differentiation of chondrocytes coincide with the proliferative arrest of the differentiating cells. However, the mechanisms by which differentiation is coordinated with cell cycle withdrawal, and the importance of this coordination for skeletal development, have not been defined. Through analysis of mice lacking the pRB-related p107 and p130 proteins, we found that p107 was required in prechondrogenic condensations for cell cycle withdrawal and for quantitatively normal alpha1(II) collagen expression. Remarkably, the p107-dependent proliferative arrest of mesenchymal cells was not needed for qualitative changes that are associated with chondrogenic differentiation, including production of Alcian blue-staining matrix and expression of the collagen IIB isoform. In chondrocytes, both p107 and p130 contributed to cell cycle exit, and p107 and p130 loss was accompanied by deregulated proliferation, reduced expression of Cbfa1, and reduced expression of Cbfa1-dependent genes that are associated with hypertrophic differentiation. Moreover, Cbfa1 was detected, and hypertrophic differentiation occurred, only in chondrocytes that had undergone or were undergoing a proliferative arrest. The results suggest that Cbfa1 links a p107- and p130-mediated cell cycle arrest to chondrocyte terminal differentiation. PMID- 12086467 TI - Role of PTPase(s) in regulating Fyn kinase at fertilization of the zebrafish egg. AB - Fertilization involves the activation of Src-family protein kinases which play a role at multiple stages of the egg activation process. The objective of the present study was to determine the mechanism by which one of these kinases, the Fyn kinase, is activated in response to fertilization of the zebrafish egg. Inhibitor studies demonstrated that many aspects of egg activation, including Fyn activation, require phosphotyrosyl phosphatase activity. A phosphotyrosyl phosphatase was found to be tightly associated with Fyn kinase and this interaction was mapped to the SH2 domain of Fyn. Coimmunoprecipitation studies identified rPTPalpha as a phosphatase that is complexed with Fyn in the egg, raising the possibility that rPTPalpha is part of the regulatory mechanism responsible for activating Fyn at fertilization. PMID- 12086468 TI - A Role for phosphoinositide 3-kinase in the control of cell division and survival during retinal development. AB - Neurogenesis in the retina requires the concerted action of three different cellular processes: proliferation, differentiation, and apoptosis. Class IA phosphoinositide 3-kinase (PI3K) is a heterodimer composed of a p85 regulatory and a p110 catalytic subunit. p110alpha has been shown to regulate cell division and survival. Little is known of its function in development, however, as p110alpha knockout mice exhibit CNS defects, but death at early embryonic stages impairs further study. Here, we examine the role of PI3K in mouse retina development by expressing an activating form of PI3K regulatory subunit, p65(PI3K), as a transgene in the retina. Mice expressing p65(PI3K) showed severely disrupted retina morphogenesis, with ectopic cell masses in the neuroepithelium that evolved into infoldings of adult retinal cell layers. These changes correlated with an altered cell proliferation/cell death balance at early developmental stages. Nonetheless, the most affected cell layer in adult retina was that of photoreceptors, which correlated with selectively increased survival of these cells at developmental stages at which cell division has ceased. These results demonstrate the relevance of accurate PI3K regulation for normal retinal development, supporting class IA PI3K involvement in induction of cell division at early stages of neurogenesis. These data also show that, even after cell division decline, PI3K activation mediates survival of differentiated neurons in vivo. PMID- 12086469 TI - Experimental studies on the spatiotemporal expression of WT1 and RALDH2 in the embryonic avian heart: a model for the regulation of myocardial and valvuloseptal development by epicardially derived cells (EPDCs). AB - Epicardially derived cells (EPDCs) delaminate from the primitive epicardium through an epithelial-to-mesenchymal transformation (EMT). After this transformation, a subpopulation of cells progressively invades myocardial and valvuloseptal tissues. The first aim of the study was to determine the tissue specific distribution of two molecules that are thought to play a crucial function in the interaction between EPDCs and other cardiac tissues, namely the Wilms' Tumor transcription factor (WT1) and retinaldehyde-dehydrogenase2 (RALDH2). This study was performed in normal avian and in quail-to-chick chimeric embryos. It was found that EPDCs that maintain the expression of WT1 and RALDH2 initially populate the subepicardial space and subsequently invade the ventricular myocardium. As EPDCs differentiate into the smooth muscle and endothelial cell lineage of the coronary vessels, the expression of WT1 and RALDH2 becomes downregulated. This process is accompanied by the upregulation of lineage-specific markers. We also observed EPDCs that continued to express WT1 (but very little RALDH2) which did not contribute to the formation of the coronary system. A subset of these cells eventually migrates into the atrioventricular (AV) cushions, at which point they no longer express WT1. The WT1/RALDH2-negative EPDCs in the AV cushions do, however, express the smooth muscle cell marker caldesmon. The second aim of this study was to determine the impact of abnormal epicardial growth on cardiac development. Experimental delay of epicardial growth distorted normal epicardial development, reduced the number of invasive WT1/RALDH2-positive EPDCs, and provoked anomalies in the coronary vessels, the ventricular myocardium, and the AV cushions. We suggest that the proper development of ventricular myocardium is dependent on the invasion of undifferentiated, WT1-positive, retinoic acid-synthesizing EPDCs. Furthermore, we propose that an interaction between EPDCs and endocardial (derived) cells is imperative for correct development of the AV cushions. PMID- 12086470 TI - Infertility of CD9-deficient mouse eggs is reversed by mouse CD9, human CD9, or mouse CD81; polyadenylated mRNA injection developed for molecular analysis of sperm-egg fusion. AB - CD9 is a membrane protein belonging to the tetraspanin family. Despite CD9's broad tissue distribution, the only abnormality observed in CD9-deficient mice was infertility of females, which was responsible for a defect in the sperm-egg fusion process. However, the function of CD9 in sperm-egg fusion is not clear at all because the technique to analyze the activity of molecules in sperm-egg fusion has not been established. We demonstrated that the exogenous mouse CD9, expressed by polyadenylated mRNA injection at the germinal-vesicle stage oocytes, was precisely localized to the egg plasma membrane, and the expression reversed the infertility of CD9(-/-) eggs. Then, two other tetraspanins, human CD9 and mouse CD81, overexpressed with this technique on CD9(-/-) eggs restored the fertilization rate up to approximately 90 and approximately 50% against that of wild type eggs, respectively. Moreover, in the presence of an anti-mouse CD9 mAb, which blocks sperm-egg fusion, expression of human CD9 or mouse CD81 on eggs also rescued the fusibility. These results suggested that human CD9 plays a crucial role in human fertilization, and mouse CD81 has the potential to compensate for CD9 function in sperm-egg fusion. In addition, the polyadenylated mRNA injection is effective for molecular analysis of sperm-egg fusion. PMID- 12086471 TI - Pattern formation in a pentameral animal: induction of early adult rudiment development in sea urchins. AB - We investigated adult rudiment induction in the direct-developing sea urchin Heliocidaris erythrogramma microsurgically. After removal of the archenteron (which includes presumptive coelomic mesoderm as well as presumptive endoderm) from late gastrulae, larval ectoderm develops properly but obvious rudiments (tube feet, nervous system, and adult skeleton) fail to form, indicating that coelomic mesoderm, endoderm, or both are required for induction of adult development. Recombination of ectoderm and archenteron rescues development. Implanted endoderm alone or left coelom alone each regenerate the full complement of archenteron derivatives; thus, they are uninformative as to the relative inductive potential of the two regions. However, in isolated ectoderm, more limited regeneration gives rise to larvae containing no archenteron derivatives at all, endoderm only, or both endoderm and left coelom. Adult nervous system begins to develop only in the latter, indicating that left coelom is required for the inductive signal. Isolated ectoderm develops a vestibule (the precursor of adult ectoderm) and correctly regulates vestibular expression of the ectodermal territory marker HeET-1, indicating that the early phase of vestibule development occurs autonomously; only later development requires the inductive signal. Another ectodermal marker, HeARS, is regulated properly in the larval ectoderm region, but not in the vestibule. HeARS regulation thus represents an early response to the inducing signal. We compare HeARS expression in H. erythrogramma with that in indirect developers and discuss its implications for modularity in the evolution of developmental mode. PMID- 12086472 TI - Fgf4 positively regulates scleraxis and tenascin expression in chick limb tendons. AB - In vertebrates, tendons connect muscles to skeletal elements. Surgical experiments in the chick have underlined developmental interactions between tendons and muscles. Initial formation of tendons occurs autonomously with respect to muscle. However, further tendon development requires the presence of muscle. The molecular signals involved in these interactions remain unknown. In the chick limb, Fgf4 transcripts are located at the extremities of muscles, where the future tendons will attach. In this paper, we analyse the putative role of muscle-Fgf4 on tendon development. We have used three general tendon markers, scleraxis, tenascin, and Fgf8 to analyse the regulation of these tendon associated molecules by Fgf4 under different experimental conditions. In the absence of Fgf4, in muscleless and aneural limbs, the expression of the three tendon-associated molecules, scleraxis, tenascin, and Fgf8, is down-regulated. Exogenous implantation of Fgf4 in normal, aneural, and muscleless limbs induces scleraxis and tenascin expression but not that of Fgf8. These results indicate that Fgf4 expressed in muscle is required for the maintenance of scleraxis and tenascin but not Fgf8 expression in tendons. PMID- 12086473 TI - Zebrafish hox paralogue group 2 genes function redundantly as selector genes to pattern the second pharyngeal arch. AB - The pharyngeal arches are one of the defining features of the vertebrates, with the first arch forming the mandibles of the jaw and the second forming jaw support structures. The cartilaginous elements of each arch are formed from separate migratory neural crest cell streams, which derive from the dorsal aspect of the neural tube. The second and more posterior crest streams are characterized by specific Hox gene expression. The zebrafish has a larger overall number of Hox genes than the tetrapod vertebrates, as the result of a duplication event in its lineage. However, in both zebrafish and mouse, there are just two members of Hox paralogue group 2 (PG2): Hoxa2 and Hoxb2. Here, we show that morpholino-mediated "knock-down" of both zebrafish Hox PG2 genes results in major defects in second pharyngeal arch cartilages, involving replacement of ventral elements with a mirror-image duplication of first arch structures, and accompanying changes to pharyngeal musculature. In the mouse, null mutants of Hoxa2 have revealed that this single Hox gene is required for normal second arch patterning. By contrast, loss-of-function of either zebrafish Hox PG2 gene individually has no phenotypic consequence, showing that these two genes function redundantly to confer proper pattern to the second pharyngeal arch. We have also used hoxb1a mis-expression to induce localized ectopic expression of zebrafish Hox PG2 genes in the first arch; using this strategy, we find that ectopic expression of either Hox PG2 gene can confer second arch identity onto first arch structures, suggesting that the zebrafish Hox PG2 genes act as "selector genes." PMID- 12086474 TI - Structure and expression of three Emx genes in the dogfish Scyliorhinus canicula: functional and evolutionary implications. AB - We report the characterization of three Emx genes in a chondrichthyan, the dogfish Scyliorhinus canicula. Comparisons of these genes with their osteichthyan counterparts indicate that the gnathostome Emx genes belong to three distinct orthology classes, each containing one of the dogfish genes and either the tetrapod Emx1 genes (Emx1 class), the osteichthyan Emx2 genes (Emx2 class) or the zebrafish Emx1 gene (Emx3 class). While the three classes could be retrieved from the pufferfish genome data, no indication of an Emx3-related gene in tetrapods could be found in the databases, suggesting that this class may have been lost in this taxon. Expression pattern comparisons of the three dogfish Emx genes and their osteichthyan counterparts indicate that not only telencephalic, but also diencephalic Emx expression territories are highly conserved among gnathostomes. In particular, all gnathostomes share an early, dynamic phase of Emx expression, spanning presumptive dorsal diencephalic territories, which involves Emx3 in the dogfish, but another orthology class, Emx2, in tetrapods. In addition, the dogfish Emx2 gene shows a highly specific expression domain in the cephalic paraxial mesoderm from the end of gastrulation and throughout neurulation, which suggests a role in the segmentation of the cephalic mesoderm. PMID- 12086476 TI - Novel 2-oxoamide inhibitors of human group IVA phospholipase A(2). AB - A novel class of potent human cytosolic phospholipase A(2) (GIVA PLA(2)) inhibitors was developed. These inhibitors were designed to contain the 2 oxoamide functionality and a free carboxyl group. Among the compounds tested, a long-chain 2-oxoamide containing L-gamma-norleucine was the most potent inhibitor, causing a 50% decrease in GIVA PLA(2) activity at 0.009 mole fraction. PMID- 12086475 TI - Naphthalene dicarboxaldehyde as an electrophilic fluorogenic moiety for affinity labeling: application to opioid receptor affinity labels with greatly improved fluorogenic properties. AB - To develop ligands with fluorogenic properties amenable for following the kinetics of cross-linking to receptors, a naphthalene dicarboxaldehyde moiety has been attached to an opiate pharmacophore 2 and evaluated in mu-opioid receptors. The fluorescence of the benzo[f]isoindole formed upon cross-linking of mu-opioid receptors by 2 permitted the time-course of covalent bonding to be followed. This demonstrated proof-of-concept suggests the usefulness of naphthalene dicarboxaldehyde-containing affinity labels as kinetic probes. PMID- 12086477 TI - An investigation of bone resorption and Dictyostelium discoideum growth inhibition by bisphosphonate drugs. AB - We report the results of 3D-QSAR/CoMFA investigations of the activity of bisphosphonate drugs, farnesyl pyrophosphate synthase (FPPSase) inhibitors, in the inhibition of bone resorption as well as the growth of Dictyostelium discoideum. In the case of D. discoideum, we find an experimental versus QSAR predicted pIC(50) R(2) value of 0.94 for 16 bisphosphonates over the 9-1200 microM range of IC(50) values, a cross-validated R(2) = 0.90, and a bootstrapped R(2) = 0.94, and we demonstrate that this approach has predictive utility (a 0.18 pIC(50) rms error for three test sets of 3 predictions). In bone resorption, we find an experimental versus predicted pLED (lowest effective dose) R(2) = 0.79 for 35 bisphosphonates over the 0.0001-1 mg of P/kg LED range, a cross-validated R(2) = 0.75, and a bootstrapped R(2) = 0.79. Two sets of 31 compounds were used as training sets for the predicted pLED values for two sets of 4 compounds which have an rms error of 0.44, larger than that found with D. discoideum. However, this can be attributed to the rather large uncertainties in the experimental bone resorption data which are almost all reported in decade steps (DeltapLED = 1). The CoMFA predicted (rat) bone antiresorptive pLED values are in agreement with literature (human recombinant) FPPSase inhibition results with an rms error of 0.45 (a factor of 2.8 error in activity prediction). We also report the single crystal X-ray crystallographic structure of the compound most active in D. discoideum growth inhibition, 2-(3-picolyl)-aminomethylene-1,1-bisphosphonic acid. The structure clearly shows the presence of bond length alternation in the picolyl ring and a planar amino group linked by a very short (1.346 A) bond to the picolyl group, an amidinium-like structure which is also expected to occur in other highly active species such as minodronate and zoledronate. Overall, these results show that it is now possible to predict the activity of bisphosphonates using 3D-QSAR/CoMFA methods, although bone resorption studies should benefit from additional, accurate information on enzyme inhibition. PMID- 12086478 TI - Activity of bisphosphonates against Trypanosoma brucei rhodesiense. AB - We report the results of a comparative molecular field analysis (CoMFA) investigation of the growth inhibition of the bloodstream form of Trypanosoma brucei rhodesiense trypomastigotes by bisphosphonates. A quantitative three dimensional structure-activity relationship CoMFA model for a set of 26 bisphosphonates having a range of activity spanning approximately 3 orders of magnitude (minimum IC(50) = 220 nM; maximum IC(50) = 102 microM) yielded an R(2) value of 0.87 with a cross-validated R(2) value of 0.79. The predictive utility of this approach was tested for three sets of three compounds: the average pIC(50) error was 0.23. For the nitrogen-containing bisphosphonates, in general, the activity was aromatic- >> aliphatic-containing side chains. The activity of aromatic species lacking an alkyl ring substitution decreased from ortho to meta to para substitution; halogen substitutions also reduced activity. For the aliphatic bisphosphonates, the IC(50) values decreased nearly monotonically with increasing chain length (down to IC(50) = 2.0 microM for the n-C(11) alkyl side chain species). We also show, using a "rescue" experiment, that the molecular target of the nitrogen-containing bisphosphonate, risedronate, in T. b. rhodesiense is the enzyme farnesyl pyrophosphate synthase. In addition, we report the LD(50) values of bisphosphonates in a mammalian cell general toxicity screen and present a comparison between the therapeutic indices and the IC(50) values in the T. b. rhodesiense growth inhibition assay. Several bisphosphonates were found to have large therapeutic indices (> or =200:1) as well as low IC(50) values, suggesting their further investigation as antiparasitic agents against T. b. rhodesiense. PMID- 12086479 TI - Principles governing the binding of a class of non-peptidic inhibitors to the SH2 domain of src studied by X-ray analysis. AB - A total of 11 structures of the (pp60)src SH2 domain with non-peptidic inhibitors based on the same two closely related inhibitor scaffolds were determined using X ray crystallography. Surprisingly, the inhibitors that have an IC(50) value between 4 and 2700 nM bind in three different binding modes. Structure comparisons show that the inhibitors aim to maximize the interaction between the hydrophobic substituent and the hydrophobic pY+3 pocket. This is achieved either by an alternative binding mode of the phenyl phosphate or by including water molecules that mediate the interaction between the inhibitor scaffold and a rigid surface of the SH2 domain. The combination of the rigid pY+3 pocket and the rigid protein surface to which the scaffolds bind results in severe distance and angular restraints for putative scaffolds and their substituents. The X-ray data suggest that these restraints seem to be compensated in our system by including water molecules, thereby increasing the flexibility of the system. PMID- 12086480 TI - Structural requirements for inhibition of the neuronal nitric oxide synthase (NOS I): 3D-QSAR analysis of 4-oxo- and 4-amino-pteridine-based inhibitors. AB - The family of homodimeric nitric oxide synthases (NOS I-III) catalyzes the generation of the cellular messenger nitric oxide (NO) by oxidation of the substrate L-arginine. The rational design of specific NOS inhibitors is of therapeutic interest in regulating pathological NO levels associated with sepsis, inflammatory, and neurodegenerative diseases. The cofactor (6R)-5,6,7,8 tetrahydrobiopterin (H(4)Bip) maximally activates all NOSs and stabilizes enzyme quaternary structure by promoting and stabilizing dimerization. Here, we describe the synthesis and three-dimensional (3D) quantitative structure-activity relationship (QSAR) analysis of 65 novel 4-amino- and 4-oxo-pteridines (antipterins) as inhibitors targeting the H(4)Bip binding site of the neuronal NOS isoform (NOS-I). The experimental binding modes for two inhibitors complexed with the related endothelial NO synthase (NOS-III) reveal requirements of biological affinity and form the basis for ligand alignment. Different alignment rules were derived by building other compounds accordingly using manual superposition or a genetic algorithm for flexible superposition. Those alignments led to 3D-QSAR models (comparative molecular field analysis (CoMFA) and comparative molecular similarity index analysis (CoMSIA)), which were validated using leave-one-out cross-validation, multiple analyses with two and five randomly chosen cross-validation groups, perturbation of biological activities by randomization or progressive scrambling, and external prediction. An iterative realignment procedure based on rigid field fit was used to improve the consistency of the resulting partial least squares models. This led to consistent and highly predictive 3D-QSAR models with good correlation coefficients for both CoMFA and CoMSIA, which correspond to experimentally determined NOS-II and -III H(4)Bip binding site topologies as well as to the NOS-I homology model binding site in terms of steric, electrostatic, and hydrophobic complementarity. These models provide clear guidelines and accurate activity predictions for novel NOS-I inhibitors. PMID- 12086481 TI - 4,5-diphenyltriazol-3-ones: openers of large-conductance Ca(2+)-activated potassium (maxi-K) channels. AB - A series of diphenyl-substituted heterocycles were synthesized and evaluated by electrophysiological techniques as openers of the cloned mammalian large conductance, Ca(2+)-activated potassium (maxi-K) channel. The series was designed from deannulation of known benzimidazolone maxi-K opener NS-004 (2) thereby providing an effective template for obtaining structure-activity-related information. The triazolone ring system was the most studied wherein 4,5 diphenyltriazol-3-one 6d (maxi-K = 158%) was identified as the optimal maxi-K channel opener. PMID- 12086482 TI - New p-methylsulfonamido phenylethylamine analogues as class III antiarrhythmic agents: design, synthesis, biological assay, and 3D-QSAR analysis. AB - Class III antiarrhythmic agents selectively delay the effective refractory period (ERP) and increase the transmembrane action potential duration (APD). Using dofetilide (2) as a template of class III antiarrhythmic agents, we designed and synthesized 16 methylsulfonamido phenylethylamine analogues (4a-d and 5a-l). Pharmacological assay indicated that all of these compounds showed activity for increasing the ERP in isolated animal atrium; among them, the effective concentration of compound 4a is 1.6 x 10(-8) mol/L in increasing ERP by 10 ms, slightly less potent than that of 2, 1.1 x 10(-8) mol/L. Compound 4a also produced a slightly lower change in ERP at 10(-5) M, DeltaERP% = 17.5% (DeltaERP% = 24.0% for dofetilide). On the basis of this bioassay result, these 16 compounds together with dofetilide were investigated by the three-dimensional quantitative structure-activity relationship (3D-QSAR) techniques of comparative molecular field analysis (CoMFA), comparative molecular similarity index analysis (CoMSIA), and the hologram QSAR (HQSAR). The 3D-QSAR models were tested with another 11 compounds (4e-h and 5m-s) that we synthesized later. Results revealed that the CoMFA, CoMSIA, and HQSAR predicted activities for the 11 newly synthesized compounds that have a good correlation with their experimental value, r(2) = 0.943, 0.891, and 0.809 for the three QSAR models, respectively. This indicates that the 3D-QSAR models proved a good predictive ability and could describe the steric, electrostatic, and hydrophobic requirements for recognition forces of the receptor site. On the basis of these results, we designed and synthesized another eight new analogues of methanesulfonamido phenylethyamine (6a-h) according to the clues provided by the 3D-QSAR analyses. Pharmacological assay indicated that the effective concentrations of delaying the ERP by 10 ms of these newly designed compounds correlated well with the 3D-QSAR predicted values. It is remarkable that the percent change of delaying ERP at 10(-5) M compound 6c is much higher than that of dofetilide; the effective concentration of compound 6c is 5.0 x 10( 8)mol/L in increasing the ERP by 10 ms, which is slightly lower than that of 2. The results showed that the 3D-QSAR models are reliable and can be extended to design new antiarrhythmic agents. PMID- 12086483 TI - Prediction of activity for nonnucleoside inhibitors with HIV-1 reverse transcriptase based on Monte Carlo simulations. AB - Results of Monte Carlo (MC) simulations for more than 200 nonnucleoside inhibitors of HIV-1 reverse transcriptase (NNRTIs) representing eight diverse chemotypes have been correlated with their anti-HIV activities in an effort to establish simulation protocols and methods that can be used in the development of more effective drugs. Each inhibitor was modeled in a complex with the protein and by itself in water, and potentially useful descriptors of binding affinity were collected during the MC simulations. A viable regression equation was obtained for each data set using an extended linear response approach, which yielded r(2) values between 0.54 and 0.85 and an average unsigned error of only 0.50 kcal/mol. The most common descriptors confirm that a good geometrical match between the inhibitor and the protein is important and that the net loss of hydrogen bonds with the inhibitor upon binding is unfavorable. Other physically reasonable descriptors of binding are needed on a chemotype case-by-case basis. By including descriptors in common from the individual fits, combination regressions that include multiple data sets were also developed. This procedure led to a refined "master" regression for 210 NNRTIs with an r(2) of 0.60 and a cross-validated q(2) of 0.55. The computed activities show an rms error of 0.86 kcal/mol in comparison with experiment and an average unsigned error of 0.69 kcal/mol. Encouraging results were obtained for the predictions of 27 NNRTIs, representing a new chemotype not included in the development of the regression model. Predictions for this test set using the master regression yielded a q(2) value of 0.51 and an average unsigned error of 0.67 kcal/mol. Finally, additional regression analysis reveals that use of ligand-only descriptors leads to models with much diminished predictive ability. PMID- 12086484 TI - Identification of potent and novel alpha4beta1 antagonists using in silico screening. AB - The antigen alpha4beta1 (very late antigen-4, VLA-4) plays an important role in the migration of white blood cells to sites of inflammation. It has been implicated in the pathology of a variety of diseases including asthma, multiple sclerosis, and rheumatoid arthritis. We describe a series of potent inhibitors of alpha4beta1 that were discovered using computational screening for replacements of the peptide region of an existing tetrapeptide-based alpha4beta1 inhibitor (1; 4-[N'-(2-methylphenyl)ureido]phenylacetyl-Leu-Asp-Val) derived from fibronectin. The search query was constructed using a model of 1 that was based upon the X-ray conformation of the related integrin-binding region of vascular cell adhesion molecule-1 (VCAM-1). The 3D search query consisted of the N-terminal cap and the carboxyl side chain of 1 because, upon the basis of existing structure-activity data on this series, these were known to be critical for high-affinity binding to alpha4beta1. The computational screen identified 12 reagents from a virtual library of 8624 molecules as satisfying the model and our synthetic filters. All of the synthesized compounds tested inhibit alpha4beta1 association with VCAM-1, with the most potent compound having an IC(50) of 1 nM, comparable to the starting compound. Using CATALYST, a 3D QSAR was generated that rationalizes the variation in activities of these alpha4beta1 antagonists. The most potent compound was evaluated in a sheep model of asthma, and a 30 mg nebulized dose was able to inhibit early and late airway responses in allergic sheep following antigen challenge and prevented the development of nonspecific airway hyperresponsiveness to carbachol. Our results demonstrate that it is possible to rapidly identify nonpeptidic replacements of integrin peptide antagonists. This approach should be useful in identification of nonpeptidic alpha4beta1 inhibitors with improved pharmacokinetic properties relative to their peptidic counterparts. PMID- 12086485 TI - Pyrazole urea-based inhibitors of p38 MAP kinase: from lead compound to clinical candidate. AB - We report on a series of N-pyrazole, N'-aryl ureas and their mode of binding to p38 mitogen activated protein kinase. Importantly, a key binding domain that is distinct from the adenosine 5'-triphoshate (ATP) binding site is exposed when the conserved activation loop, consisting in part of Asp168-Phe169-Gly170, adopts a conformation permitting lipophilic and hydrogen bonding interactions between this class of inhibitors and the protein. We describe the correlation of the structure activity relationships and crystallographic structures of these inhibitors with p38. In addition, we incorporated another binding pharmacophore that forms a hydrogen bond at the ATP binding site. This modification affords significant improvements in binding, cellular, and in vivo potencies resulting in the selection of 45 (BIRB 796) as a clinical candidate for the treatment of inflammatory diseases. PMID- 12086486 TI - Novel, non-acylguanidine-type Na(+)/H(+) exchanger inhibitors: synthesis and pharmacology of 5-tetrahydroquinolinylidene aminoguanidine derivatives. AB - In the course of our research into new types of non-acylguanidine Na(+)/H(+) exchanger (NHE) inhibitors, we designed and synthesized aryl-fused tetrahydropyranylidene and cyclohexylidene aminoguanidine derivatives I (X = O, CH(2)), which were tested for their inhibitory effects on rat platelet NHEs. After optimization, we found that the S isomer of tetrahydroquinoline derivatives that possess a methyl group in the 4-position and a halogen or methyl group in the o-position of Ar(2) exhibited high inhibitory activity. In these compounds, (5E,7S)-[[7-(5-fluoro-2-methylphenyl)-4-methyl-7,8-dihydro-5(6H) quinolinylidene]amino]guanidine dimethanesulfonate (18, T-162559) was found to be a potent inhibitor of both rat and human platelet NHEs, with IC(50) values of 14 and 13 nM, respectively. Furthermore, in a rat myocardial infarction model in vivo (1 h ischemia-24 h reperfusion), 18 (0.1 mg/kg, intravenously administered 5 min or 2 h before coronary occlusion) showed significant activity (33% or 23% inhibition, respectively). These results suggested that 18 may exhibit a potent and long-lasting protective activity against cardiac injuries induced by ischemia reperfusion. PMID- 12086487 TI - Further characterization of structural requirements for ligands at the dopamine D(2) and D(3) receptor: exploring the thiophene moiety. AB - The present study describes the synthesis and in vitro pharmacology of a novel series of dopaminergic agents in which the classical phenylethylamine pharmacophore is replaced by a thienylethylamine moiety. In general, the novel compounds showed a moderate affinity for the dopamine (DA) D(2) and D(3) receptors. When the thienylethylamine moiety is fixed in a rigid system, the affinity for the DA receptor is significantly increased. However, in the tricyclic hexahydrothianaphthoxazine structure, the affinity for the DA receptors is diminished. PMID- 12086488 TI - Highly antibacterial active aminoacyl penicillin conjugates with acylated bis catecholate siderophores based on secondary diamino acids and related compounds. AB - New acylated bis-catecholates and 1,3-benzoxazine-2,4-dione derivatives based on secondary diamino acids (N-(aminoalkyl)glycines, N-aminopropyl-alanine, and N aminopropyl-4-aminovaleric acid), on N-(aminoalkyl)aminomethyl benzoic acids, on N-(aminoalkyl)aminomethyl phenoxyacetic acids, or on 3,5-diaminobenzoic acid were synthesized as artificial siderophores. The corresponding diamino acids were obtained from the diamines and oxocarboxylic acids by catalytic hydrogenation. The acylated bis-catecholates and 1,3-benzoxazine-2,4-diones were coupled with ampicillin or amoxicillin to new siderophore aminoacylpenicillin conjugates. These conjugates exhibited very strong antibacterial activity in vitro against Gram-negative bacterial pathogens including Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Escherichia coli, Klebsiella pneumoniae, and Serratia marcescens. The ampicillin derivative 7b (HKI 9924154) and the corresponding amoxicillin derivative 8 (HKI 9924155) represent the most active compounds. The conjugates can use bacterial iron siderophore uptake routes to penetrate the Gram-negative outer membrane permeability barrier. This was demonstrated by assays with mutants deficient in components of the iron transport systems. New siderophore penicillin V conjugates with the siderophore component attached to the phenyl ring of penicillin V are inactive against these Gram negative bacteria. PMID- 12086489 TI - Synthesis and biological evaluation at nicotinic acetylcholine receptors of N arylalkyl- and N-aryl-7-azabicyclo[2.2.1]heptanes. AB - A new series of N-arylalkyl-substituted 7-azabicyclo[2.2.1]heptanes and N-aryl substituted 7-azabicyclo[2.2.1]heptanes were synthesized and evaluated as potential ligands for neuronal nicotinic acetylcholine receptors. The in vitro binding affinities (K(i)) of the 7-azabicyclo[2.2.1]heptane derivatives were measured by inhibition of [(3)H]cytisine binding to rat brain tissue. The most potent ligand of the series was found to be N-(3-pyridylmethyl)-7 azabicyclo[2.2.1]heptane (5b, K(i) = 98 nM). The chloro analogue (5a, K(i) = 245 nM) 5a and epibatidine (1) produced dose-dependent analgesia in both hotplate and tail-flick tests when administered subcutaneously. However, when compounds 1 and 5a,b were administered intrathecally, all produced analgesia in the tail-flick test but only 5a produced analgesia in the hotplate test. PMID- 12086490 TI - Modification of the structure of a metallopeptide: synthesis and biological evaluation of (111)In-labeled DOTA-conjugated rhenium-cyclized alpha-MSH analogues. AB - Rhenium-cyclized CCMSH analogues are novel melanoma-targeting metallopeptides with high tumor uptake, long tumor retention, and low background in normal tissues, which make these metallopeptides an ideal structural motif for designing novel melanoma-targeting agents. ReCCMSH has been derivatized with a 1,4,7,10 tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) chelate so that it can be labeled with a wide variety of radionuclides for imaging and therapeutic applications. This study involved optimization of the in vivo biological properties of DOTA-ReCCMSH (S), through modification of the structure of the metallopeptide. Several DOTA-ReCCMSH analogues, Ac-Lys(DOTA)-ReCCMSH (4) DOTA ReCCMSH(Arg(11)) (6), DOTA-ReCCMSH-OH (8), and DOTA-ReCCMSH-Asp-OH (10), were synthesized using solid phase peptide synthesis followed by rhenium cyclization. The IC(50) values of the metallopeptides were determined through competitive binding assays against (125)I-(Tyr(2))-NDP. Radiolabeling of the DOTA-rhenium cyclized peptides with (111)In was carried out in NH(4)OAc (0.1 M; pH 5.5) buffered solution for 30 min at 70 degrees C. The stability of the radiolabeled complexes was evaluated in 0.01 M, pH 7.4, phosphate-buffered saline/0.1% bovine serum albumin solution. After separation of the radiolabeled peptide from the unlabeled peptide by reverse phase high-performance liquid chromatography, the biodistribution of the radiolabeled complex was performed in C57 mice bearing B16/F1 murine melanoma tumors. All radiolabeled complexes showed fast blood clearance (2 h postinjection (pi): (111)In-S, 0.07 +/- 0.03% ID/g; (111)In-4, 0.09 +/- 0.06% ID/g; (111)In-6, 0.21 +/- 0.08% ID/g; (111)In-8, 0.11 +/- 0.10% ID/g; and (111)In-10, 0.05 +/- 0.03% ID/g), and their clearance was predominantly through the urine (4 h pi: 93.5 +/- 1.7, 87.8 +/- 6.5, 89.8 +/- 4.2, 93.3 +/- 1.1, and 93.8 +/- 1.8 (% ID) for (111)In-labeled S, 4, 6, 8, and 10, respectively). Tumor uptake values of 9.45 +/- 0.90, 6.01 +/- 2.36, 17.41 +/- 5.61, 9.27 +/- 0.68, and 7.32 +/- 2.09 (% ID/g) for (111)In-labeled S, 4, 6, 8, and 10, respectively, were observed at 4 h pi. The kidney uptake was 9.27 +/- 2.65% ID/g for (111)In-S, 19.02 +/- 2.63% ID/g for (111)In-4, 7.37 +/- 1.13% ID/g for (111)In-6, 8.70 +/- 0.88% ID/g for (111)In-8, and 8.13 +/- 1.47% ID/g for (111)In-10 at 4 h pi. Complex 6 showed high melanoma uptake and lower kidney uptake than the corresponding Lys(11) analogues, supporting 6 for further investigations as a potential therapeutic radiopharmaceutical. PMID- 12086491 TI - Potent and selective inhibitors of platelet-derived growth factor receptor phosphorylation. 1. Synthesis, structure-activity relationship, and biological effects of a new class of quinazoline derivatives. AB - A new series of 4-[4-(N-substituted carbamoyl)-1-piperazinyl]-6,7 dimethoxyquinazoline derivatives were found to show potent and selective inhibition of platelet-dervied growth factor (PDGF) receptor phosphorylation. In this exploration of the structure-activity relationships (SARs) of the prototype inhibitor KN1022, the 4-nitrophenylurea moiety was probed. We found that 4 substitution on the phenyl ring was optimal and the introduction of more than two substituents on the phenyl ring decreased activities. Bulky substituents on the phenyl ring enhanced activities. Thiourea analogues were also prepared, and the SARs were found to be slightly different from those of the urea derivatives. Through this research, we obtained some potent KN1022 derivatives such as 4-(4 methylphenoxy)phenyl (36, IC(50) 0.02 micromol/L), 4-tert-butylphenyl (16, IC(50) 0.03 micromol/L), and 4-phenoxyphenyl (21, IC(50) 0.08 micromol/L) analogues, which had almost a 10-fold increase in activity against KN1022. These potent compounds retained their high selectivity against the PDGF receptor family similar to KN1022. We also observed that these compounds could inhibit the PDGF BB-induced proliferation of porcine vascular smooth muscle cells without cell toxicity almost at the same IC(50) values observed for PDGF receptor phosphorylation. To evaluate the biological effects in vivo, we selected some analogues on the basis of the measurement of the plasma drug concentration after oral administration to rats. Oral administration of the 4-chlorophenyl (6), 4 bromophenyl (9), or 4-isopropoxyphenyl (20) analogue to Sprague-Dawley rats (30 mg/kg, twice daily) resulted in significant inhibition (24-38%) of neointima formation in the carotid artery of the balloon catheter deendothelialized vessel in the rats. Therefore, 4-[4-(N-substituted carbamoyl)-1-piperazinyl]-6,7 dimethoxyquinazoline derivatives, which are potent inhibitors of PDGFR phosphorylation, may be expected to represent a new therapeutic approach for the treatment of various aspects of atherosclerosis and other cellular proliferative disorders. PMID- 12086492 TI - Lepadins D-F: antiplasmodial and antitrypanosomal decahydroquinoline derivatives from the tropical marine tunicate Didemnum sp. AB - From a new tunicate species, belonging to the genus Didemnum, three alkaloids possessing an unusual and extremely rare decahydroquinoline skeleton and showing significant and selective antiplasmodial and antitrypanosomal activity were obtained as follows: (2R*,3S*,4aR*,5R*,8aS*)-decahydro-3-hydroxy-5-(5' hydroxyoctyl)-2-methylquinoline (lepadin D,1), its quaternary nitrogen derivative (2), (2R*,2"E,3S*,4aR*,5R*,8aS*)-decahydro-3-hydroxy-5-(5'-hydroxyoctyl)-2-methyl 3-quinolinyl ester 2"-octenoic acid (lepadin E, 3), and (2S*,2"E,3S*,4aR*,5R*,8aS*)-decahydro-3-hydroxy-5-(5'-hydroxyoctyl)-2-methyl-3 quinolinyl ester 2"-octenoic acid (lepadin F, 4). These isolates may well serve as lead structures for the development of new antimalarial drugs. PMID- 12086493 TI - Structure-activity relationships of the melanocortin tetrapeptide Ac-His-DPhe-Arg Trp-NH(2) at the mouse melanocortin receptors: part 2 modifications at the Phe position. AB - The melanocortin pathway is an important participant in skin pigmentation, steroidogenesis, obesity, energy homeostasis and exocrine gland function. The centrally located melanocortin-3 and melanocortin-4 receptors (MC3R, MC4R) are involved in the metabolic and food intake aspects of energy homeostasis and are stimulated by melanocortin agonists such as alpha-melanocyte stimulation hormone (alpha-MSH). The melanocortin agonists contain the putative message sequence "His Phe-Arg-Trp," and it has been well-documented that inversion of chirality of the Phe to DPhe results in a dramatic increase in melanocortin receptor potency. Herein, we report a tetrapeptide library, based upon the template Ac-His-DPhe-Arg Trp-NH(2), consisting of 26 members that have been modified at the DPhe(7) position (alpha-MSH numbering) and pharmacologically characterized for agonist and antagonist activity at the mouse melanocortin receptors MC1R, MC3R, MC4R, and MC5R. The most notable results of this study include the identification of the tetrapeptide Ac-His-(pI)DPhe-Arg-Trp-NH(2) that is a full nanomolar agonist at the mMC1 and mMC5 receptors, a mMC3R partial agonist with potent antagonist activity (pA(2) = 7.25, K(i) = 56 nM) and, but unexpectedly, is a potent agonist at the mMC4R (EC(50) = 25 nM). This ligand possesses novel melanocortin receptor pharmacology, as compared to previously reported peptides, and is potentially useful for in vivo studies to differentiate MC3R vs MC4R physiological roles in animal models, such as primates, where "knockout" animals are not viable options. The DNal(2') substitution for DPhe resulted in a mMC3R partial agonist with antagonist activity (pA(2) = 6.5, K(i) = 295 nM) and a mMC4R (pA(2) = 7.8, K(i) = 17 nM) antagonist possessing 60- and 425-fold decreased potency, respectively, as compared with SHU9119 at these receptors. Examination of this DNal(2')-containing tetrapeptide at the F254S and F259S mutant mMC4Rs resulted in agonist activity of this mMC4R tetrapeptide antagonist, similar to that observed for the SHU9119 peptide, supporting our previously proposed hypothesis that the Phe 254 and 259 transmembrane six receptor residues are important for differentiating melanocortin sequence-based MC4R antagonists vs the agouti-related protein (AGRP) sequence-based antagonists. PMID- 12086494 TI - Data analysis of high-throughput screening results: application of multidomain clustering to the NCI anti-HIV data set. AB - The routine use of high-throughput screening (HTS) systems in the drug discovery process has resulted in an increasing need for fast, reliable analysis of massive amounts of data. A new automated multidomain clustering method that thoroughly analyzes screening data sets is used to examine both the active and the inactive compounds in a well-known, publicly available data set based on primary screening. Large and small compound sets that defined both chemical families and potential pharmacophore points were discovered. The detection of structure activity relationships (SAR), aided by the unique classification method, is described in this article. PMID- 12086495 TI - Synthesis and pharmacology of benzoxazines as highly selective antagonists at M(4) muscarinic receptors. AB - Previously, we reported on PD 102807 (41) as being the most selective synthetic M(4) muscarinic antagonist identified to date. Synthesized analogues of 41 showed no improvement in affinity and selectivity at that time. However, several newly synthesized compounds exhibit a 7-fold higher affinity at M(4) receptors and demonstrate a selectivity of at least 100-fold over all other muscarinic receptor subtypes. For example, compound 28 showed an affinity of pK(i) = 9.00 at M(4) receptors and a selectivity of M(1)/M(4) = 13 183-fold, M(2)/M(4) = 339-fold, M(3)/M(4) = 151-fold, and M(5)/M(4) = 11 220-fold. This high selectivity along with high affinity has not been reported for any synthetic muscarinic antagonist, nor for natural occurring M(4) antagonists such as the M(4) selective Eastern Green Mamba venom MT3 (M(4) pK(b) = 8.7, M(1)/M(4) = 40-fold, M(2)/M(4) > or = 500-fold, M(3)/M(4) > or = 500-fold, and M(5)/M(4) > or = 500-fold). Derivative 24, a compound with a high selectivity pattern as well, has been tested for in vivo efficacy. It was able to block the L-3,4-dihydroxyphenylalanine accumulation produced by CI-1017, an M(1)/M(4) selective muscarinic agonist, in the mesolimbic region and striatum, which suggests that 24 is capable of crossing the blood brain barrier and confirms the pharmacokinetic data obtained on this compound. This is evidence that suggests that agonist-induced increase in catecholamine synthesis observed in these regions is mediated by M(4) receptors. PMID- 12086496 TI - Correlations between cytotoxicity and topography of some 2 arylidenebenzocycloalkanones determined by X-ray crystallography. AB - Three series of 2-arylidenebenzocycloalkanones 1-3 were prepared in order to compare the topography of the molecules with cytotoxicity. These compounds contain two aryl rings whose spatial relationships to each other were influenced by the size of the alicyclic ring and the nature of the substituents in the arylidene aryl rings. All compounds were evaluated against murine P388 and L1210 cells as well as human Molt 4/C8 and CEM T-lymphocytes. From these results, 1l and 2c,l emerged as useful lead molecules and 1l was shown to significantly inhibit macromolecular DNA, RNA, and protein syntheses in L1210 cells. Various interatomic distances, bond angles, and a torsion angle of 19 representative compounds were determined by X-ray crystallography, and correlations between these data and the cytotoxicity were noted in nearly 40% of the cases examined. Structure-activity relationships revealed that in general, the steric properties of the groups in the arylidene aryl ring, as revealed by measurements of the molar refractivity values, contributed more to bioactivity than the electronic and hydrophobic properties of the aryl substituents. The compounds displayed little murine toxicity, which favors the decision to develop these molecules as cytotoxic and anticancer agents. PMID- 12086497 TI - Potent, novel in vitro inhibitors of the Pseudomonas aeruginosa deacetylase LpxC. AB - Deacetylation of uridyldiphospho-3-O-(R-hydroxydecanoyl)-N-acetylglucosamine by LpxC is the first committed step in the Pseudomonas aeruginosa biosynthetic pathway to lipid A; homologous enzymes are found widely among Gram-negative bacteria. As an essential enzyme for which no inhibitors have yet been reported, the P. aeruginosa LpxC represents a highly attractive target for a novel antibacterial drug. We synthesized several focused small-molecule libraries, each composed of a variable aromatic ring, one of four heterocyclic/spacer moieties, and a hydroxamic acid and evaluated the LpxC inhibition of these compounds against purified P. aeruginosa enzyme. To ensure that the in vitro assay would be as physiologically relevant as possible, we synthesized a tritiated form of the specific P. aeruginosa glycolipid substrate and measured directly the enzymatically released acetate. Several of our novel compounds, predominantly those having fluorinated substituents on the aromatic ring and an oxazoline as the heterocyclic moiety, demonstrated in vitro IC(50) values less than 1 microM. We now report the synthesis and in vitro evaluation of these P. aeruginosa LpxC inhibitors. PMID- 12086499 TI - 2-Amino-6-arylthio-9-[2-(phosphonomethoxy)ethyl]purine bis(2,2,2-trifluoroethyl) esters as novel HBV-specific antiviral reagents. AB - Novel 2-amino-6-arylthio-9-[2-(phosphonomethoxy)ethyl]purine bis(2,2,2 trifluoroethyl) esters were synthesized and evaluated for antihepatitis B virus (HBV) activity in vitro using HB611, HuH-6 cell line, stably transfected with the HBV genome. Among the compounds synthesized, 2-amino-6-phenylthio-9-[2 (phosphonomethoxy)ethyl]purine bis(2,2,2-trifluoroethyl) ester (8), 2-amino-6-(4 methoxyphenylthio)-9-[2-(phosphonomethoxy)ethyl]purine bis(2,2,2-trifluoroethyl) ester (16), 2-amino-6-(3-methoxyphenylthio)-9-[2-(phosphonomethoxy)ethyl]purine bis(2,2,2-trifluoroethyl) ester (17), and 2-amino-6-(2-methoxyphenylthio)-9-[2 (phosphonomethoxy)ethyl]purine bis(2,2,2-trifluoroethyl) ester (18) showed considerably high anti-HBV activity, as represented by IC(50) values of 0.05, 0.03, 0.04, and 0.08 microM, respectively, and exhibited low cytotoxicity, as represented by CC(50) values of more than 1000 microM. It was suggested that these compounds did not have anti-HIV activity, and compound 8 showed only weak anti-HSV-1 activity. An antiviral agent, 9-[2-(phosphonomethoxy)ethyl]adenine (PMEA), which was used as a control in the present study, showed moderate anti HBV activity, as represented by an IC(50) value of 0.2 microM. Furthermore, compound 16 was administered orally to mice at a dose of 100 mg/kg in order to examine its gastrointestinal absorbability. Consequently, the main active metabolite was observed in mouse plasma, with especially high concentrations in the liver. PMID- 12086498 TI - II. Synthesis and biological evaluation of some bioisosteres and congeners of the antitumor agent, 2-(4-[(7-chloro-2-quinoxalinyl)oxy]phenoxy)propionic acid (XK469). AB - XK469 (1) is among the most highly and broadly active antitumor agents to have been evaluated in our laboratories. Subsequent developmental studies led to the entry of (R)-(+) 1 (NSC 698215) into phase 1 clinical trials (NIH UO1-CA62487). The antitumor mechanism of action of 1 remains to be elucidated, which has prompted a sustained effort to elaborate a pharmacophoric pattern of 1. The present study focused on a strategy of synthesis and biological evaluation of topologically based, bioisosteric replacements of the quinoxaline moiety in the lead compound (1) by quinazoline (4a-d), 1,2,4-benzotriazine (12a-18b), and quinoline (21a-g) ring systems. The synthetic approach to each of the bioisosteres of 1 utilized the methodology developed in previous work (see Hazeldine, S. T.; Polin, L.; Kushner, J.; Paluch, J.; White, K.; Edelstein, M.; Palomino, E.; Corbett, T. H.; Horwitz, J. P. Design, Synthesis, and Biological Evaluation of Analogues of the Antitumor Agent 2-(4-[(7-Chloro-2 quinoxalinyl)oxy]phenoxy)propionic acid (XK469). J. Med. Chem. 2001, 44, 1758 1776.), which is extended to the procurement of the benzoxazole (23a,b), benzthiazole (23c,d), pyridine (25a,b), and pyrazine (27) congeners of 1. Only quinoline analogues, bearing a 7-halo (21a,b,d,e) or a 7-methoxy substituent (21g), showed antitumor activities (Br > Cl > CH(3)O > F approximately I), at levels comparable to or greater than the range of activities manifested by 1 and corresponding analogues. At high individual dosages, the (S)-(-) enantiomers of 1 and 21b,d all produce a reversible slowing of nerve-conduction velocity in the mice, the onset of which is characterized by a distinctive dysfunction of the hind legs, causing uncoordinated movements. The condition resolves within 5-10 min. However, at higher dosages, which approach a lethal level, the behavior extended to the front legs, lasting from 20 min to 1 h. By contrast, the (R)-(+) forms of these same agents did not induce the phenomenon of slowing of nerve conduction velocity. PMID- 12086500 TI - Synthesis, SAR, and biological evaluation of oximino-piperidino-piperidine amides. 1. Orally bioavailable CCR5 receptor antagonists with potent anti-HIV activity. AB - We previously reported the discovery of 4-[(Z)-(4 bromophenyl)(ethoxyimino)methyl]-1'-[(2,4-dimethyl-3-pyridinyl)carbonyl]-4' methyl-1,4'-bipiperidine N-oxide 1 (SCH 351125) as an orally bioavailable human CCR5 antagonist for the treatment of HIV-1 infection. Herein, we describe in detail the discovery of 1 from our initial lead compound as well as the synthesis and SAR studies directed toward optimization of substitution at the phenyl, oxime, and right-hand side amide groups in the oximino-piperidino-piperidine series. Substitutions (4-Br, 4-CF(3), 4-OCF(3), 4-SO(2)Me, and 4-Cl) at the phenyl group are well-tolerated, and small alkyl substitutions (Me, Et, (n)()Pr, (i)()Pr, and cyclopropyl methyl) at the oxime moiety are preferred for CCR5 antagonism. The 2,6-dimethylnicotinamide N-oxide moiety is the optimal choice for the right-hand side. Several compounds in this series, including compound 1, exhibited excellent antiviral activity in vitro. Compound 1, which has a favorable pharmacokinetic profile in rodents and primates, excellent oral bioavailability, and potent antiviral activity against a wide range of primary HIV-1 isolates, is a potentially promising new candidate for treatment of HIV-1 infection. PMID- 12086501 TI - Traditional Chinese medicine: a brief outline. PMID- 12086502 TI - On the number of benzenoid hydrocarbons. AB - We present a new algorithm which allows a radical increase in the computer enumeration of benzenoids b(h) with h hexagons. We obtain b(h) up to h = 35. We prove that b(h) approximately const.kappa(h), prove the rigorous bounds 4.789 < or = kappa < or = 5.905, and estimate that kappa = 5.16193016(8). Finally, we provide strong numerical evidence that the generating function summation operator b(h)z(h) approximately A(z) log(1 - kappa z), estimate A(1/kappa) and predict the subleading asymptotic behavior. We also provide compelling arguments that the mean-square radius of gyration (h) of benzenoids of size h grows as h(2 nu), with nu = 0.64115(5). PMID- 12086503 TI - Information sources on post-grant actions to pharmaceutical patents. AB - Over the past two decades, many industrialized countries have implemented changes in their patent laws to provide an extended period of protection for commercially marketed pharmaceuticals. The process has been adopted piecemeal, using widely different procedures. The information sources relating to such term extension are consequently scattered and poorly organized, compared with conventional patent legal status data. This paper reviews the legal processes behind term extension and provides a survey of the key principles and information sources available. PMID- 12086504 TI - Data quality assurance for thermophysical property databases--applications to the TRC SOURCE data system. AB - To a significant degree processes of database development are based upon human activities, which are susceptible to various errors. Propagation of errors in the processing leads to a decrease in the value of original data as well as that of any database products. Data quality is a critical issue that every database producer must handle as an inseparable part of the database management. Within the Thermodynamics Research Center (TRC), a systematic approach to implement database integrity rules was established through the use of modern database technology, statistical methods, and thermodynamic principles. The four major functions of the system--error prevention, database integrity enforcement, scientific data integrity protection, and database traceability--are detailed in this paper. PMID- 12086505 TI - A 3D structure database of components from Chinese traditional medicinal herbs. AB - This article described a 3D structure database of components extracted from Chinese Traditional Medicinal (CTM) herbs. It offers not only basic molecular properties and optimized 3D structure of the compounds but also detailed information on their herbal origin, including basic herbal category (e.g. English name, Latin name, and family), effective parts, and clinical effects. An easy to use, interactive GUI browser allows users to perform various searches via complex logical query builder. Combined with the latest network database engine (MySQL), it can achieve excellent performance under both a local network and an Internet environment. We have tested it on the design of inhibitors of NS3-NS4A protease. Results show that the structure database of components extracted from Chinese medicinal herbs can be a rich source in searching the lead compound. PMID- 12086506 TI - On the belt and moebius isomers of the coronene molecule. AB - The structure and stability of ca. 100 nonplanar constitutional isomers of the coronene molecule of general formula C(24)H(12) have been estimated by semiempirical and by ab initio calculations. The isomers studied are unbranched cyclic catacondensed structures and fall into two classes, viz., the belt (B) and the Moebius (M) type. The calculations have shown some of the M-isomers to be more stable than the corresponding B-isomers and thus to form a platform for the future synthesis of or search for such isomers in Nature. PMID- 12086507 TI - Looking for natural patterns in analytical data. 2. Tracing local density with OPTICS. AB - The main principles and the algorithm of a density-based clustering approach, OPTICS, are described, and its unique properties, such as the ability to reveal clusters of arbitrary shapes and different densities, are illustrated on simulated and real spectral and chromatographic data sets. A "reachability plot" visualizing density fluctuations of data in multivariate space and a "color map" relating the original and/or descriptive features with data clustering allow a deeper insight into the data structure and its interpretation in chemical terms. PMID- 12086508 TI - Predicting bond lengths in planar benzenoid polycyclic aromatic hydrocarbons: a chemometric approach. AB - Two hundred and twenty-three aromatic carbon-carbon bond lengths in high precision crystal structures containing 22 planar condensed benzenoid polycyclic aromatic hydrocarbons (PB-PAHs) were related to the Pauling pi-bond order, its analogue corrected to crystal packing effects, the number of hexagonal rings around the bond, and the numbers of carbons atoms around the bond at topological distance one and two. Principal Component Analysis (PCA) showed that the bond lengths in PB-PAHs are at least two-dimensional phenomenon, with well pronounced classification into 12 types of bonds, as confirmed with Hierachical Cluster Analysis (HCA). Consequently, Multiple Linear Regression (MLR) and Partial Least Squares (PLS) models were superior to univariate models, reducing the degeneration of the data set and improving the estimation of Julg's structural aromaticity index. The approximate regression models based on topological descriptors only were built for fast and easy prediction of bond lengths and bond orders in PB-PAHs. PMID- 12086509 TI - DNA solution of a graph coloring problem. AB - The graph-theoretic parameter that has probably received the most attention over the years is the chromatic number. As is well-known, the coloring problem is an NP-Complete problem. In this paper, it has been solved by means of molecular biology techniques. The algorithm is highly parallel and has satisfactory fidelity. This work shows further evidence for the ability of DNA computing to solve NP-Complete problems. PMID- 12086510 TI - Some notes on 2-D graphical representation of DNA sequence. AB - Some 2-D and 3-D graphical representations of DNA sequences have been given by Nandy, Leong and Mogenthaler, and Randic et al., which give visual characterizations of DNA sequences. In this paper, we presented a novel graphical representation of DNA sequences by taking four special vectors in 2-D Cartesian coordinate system to represent the four nucleic acid bases in DNA sequences, so that a DNA sequence is denoted on a plane by a directed walk. It is shown that the new graphical representation of DNA sequences has lower or nondegeneracy. PMID- 12086511 TI - Use of graph theory in thermodynamics of phase equilibria. AB - A concept to use graph theory for the description of phase equilibria is developed. It is shown that a specific planar graph, called the graph of state, corresponds unequivocally to a specific state of phase equilibrium. Hence, the involved problem of enumeration of different states equilibria in complex systems is simplified to the problem of enumeration of graphs of the specified type. One phase systems with three independent components can exist in two forms, normal and exotic; while the normal form is of course known also for systems with 1 and 2 components, the exotic form can only exist if the number of components is 3 or more. It can be speculated that stable quasicrystals represent such an exotic form. Assuming the occurrence of all of the thermodynamically allowed processes, the number of one-phase exotic states in systems that consist of tens of bioelements can, intuitively, be used as a measure of Earth's biodiversity. PMID- 12086512 TI - SLIPPER-2001 -- software for predicting molecular properties on the basis of physicochemical descriptors and structural similarity. AB - A new approach for predicting the lipophilicity (log P), solubility (log Sw), and oral absorption of drugs in humans (FA) is described. It is based on structural and physicochemical similarity and is realized in the software program SLIPPER 2001. Calculated and experimental values of log P, log Sw, and FA for 42 drugs were used to demonstrate the predictive power of the program. Reliable results were obtained for simple compounds, for complex chemicals, and for drugs. Thus, the principle of "similar compounds display similar properties" together with estimating incremental changes in properties by using differences in physicochemical parameters results in "structure - property " predictive models even in the absence of a precise understanding of the mechanisms involved. PMID- 12086513 TI - Differential Shannon entropy analysis identifies molecular property descriptors that predict aqueous solubility of synthetic compounds with high accuracy in binary QSAR calculations. AB - Prediction of aqueous solubility of organic molecules by binary QSAR was used as a test case for a recently introduced entropy-based descriptor selection method. Property descriptors suitable for solubility predictions were exclusively selected on the basis of Shannon entropy calculations in molecular learning sets, not taking any other information into account. Sets of only five or 10 2D descriptors with largest entropy differences between molecules above or below a defined solubility threshold yielded consistently high prediction accuracy between 80% and 90% in binary QSAR calculations, regardless of the threshold values applied. The top five descriptors with largest differential Shannon entropy (DSE) values achieved an average prediction accuracy of 88%. These findings suggest that differences in entropy and relative information content of descriptors in compared compound data sets correlate with significant differences in physical properties and support the practical relevance of entropy-based descriptor selection routines. The study also demonstrates that binary QSAR methodology can be effectively used to classify small molecules according to aqueous solubility. PMID- 12086514 TI - On the calculation of Henry's law constants of chlorinated benzenes in water from semiempirical quantum chemical methods. AB - The Henry's law constants for all 12 polychlorinated benzene congeners were calculated using semiempirical quantum chemical solvation models (SM2, SM2.1, and SM3), and their performances are discussed. The values obtained by the SM3 method are underestimated compared with the experimental values and those calculated by SM2 and SM2.1 methods. This underestimation is larger as the degree of chlorination of the benzenes increases. This bad performance of SM3 to calculate Henry's law constants is attributable to the PM3 Hamiltonian, due to its incapacity to give a good description of electron density in chlorinated benzenes. The best results, within the 95% confidence limit of error of the experimental mean, are obtained using SM2 method. The discussion includes analysis of the different contributions to the free energy of solvation, namely, polarization free energy, cavitation, dispersion, and structural rearrangement of the solvent effects as well as partial atomic charges. PMID- 12086515 TI - Molecular basis of linear free energy relationships. The nature of inductive effect in aliphatic series. AB - The nature of inductive substituent effect on the acidity of aliphatic carboxylic acids was analyzed in terms of a quantum chemical model based on the Longuet Higgins theory of proton affinity. It has been shown that the effect is generally composed of two additive components, the electrostatic one and the polarization one. The relative weights of these components depend in any given case on the type of the reaction series. Thus, for example, the substituent-induced variation of gas-phase acidity in a series of 4-substituted bicyclo[2.2.2]octane-1 carboxylic acids is due to the electrostatic component classically described by the Kirkwood-Westheimer theory. On the other hand, the gas-phase basicity in a series of aliphatic amines is primarily determined by the polarization term that in traditional description corresponds to the concept of inductive effect. PMID- 12086516 TI - Three methods for calculation of the hyper-Wiener index of molecular graphs. AB - The hyper-Wiener index WW of a graph G is defined as WW(G) = (summation operator d (u, v)(2) + summation operator d (u, v))/2, where d (u, v) denotes the distance between the vertices u and v in the graph G and the summations run over all (unordered) pairs of vertices of G. We consider three different methods for calculating the hyper-Wiener index of molecular graphs: the cut method, the method of Hosoya polynomials, and the interpolation method. Along the way we obtain new closed-form expressions for the WW of linear phenylenes, cyclic phenylenes, poly(azulenes), and several families of periodic hexagonal chains. We also verify some previously known (but not mathematically proved) formulas. PMID- 12086517 TI - Generalized topological spaces in evolutionary theory and combinatorial chemistry. AB - The search spaces in combinatorial chemistry as well as the sequence spaces underlying (molecular) evolution are conventionally thought of as graphs. Recombination, however, implies a nongraphical structure of the combinatorial search spaces. These structures, and their implications for search process itself, are heretofore not well understood in general. In this contribution we review a very general formalism from point set topology and discuss its application to combinatorial search spaces, fitness landscapes, evolutionary trajectories, and artificial chemistries. PMID- 12086518 TI - Autocorrelation method adapted to generate new atomic environments: application for the prediction of 13-C chemical shifts of alkanes. AB - The concept of the multifunctional autocorrelation method governing global description of molecules was changed in order to take into account the structural environment of each atom. New atomic environments are generated as possible descriptors in QSARs and can be useful for database characterization. The principles of this approach are widely explained through a case study dealing with the design of a model allowing the simulation of the carbon-13 nuclear magnetic spectra for alkanes. Carbon atoms in alkanes are described by using as structural descriptors a vector corresponding to only four components vectors of the multifunctional autocorrelation method. The statistical method used for deriving the model was a classical three-layer feedforward neural network trained by the back-propagation algorithm and multilinear regression (MLR). The predictive ability of the ANN model was tested by -10%-out(L10%O) cross validation method, demonstrating the superior quality of the neural model. The established model allows us the prediction of the 13-C chemical shifts with success because since all types of carbons are taken into account without distinction of connectivity. The neural network possessed a 4:7:1 architecture with a sigmoid shape as a activation function. The model produced a cross validation standard coefficient r between delta(exp) and delta(calc) about 0.99, while the cross-validation standard s and the mean error are equal to 0.87 and 0.60 ppm, respectively. PMID- 12086519 TI - Quantitative prediction of liquid chromatography retention of N benzylideneanilines based on quantum chemical parameters and radial basis function neural network. AB - Based on quantum chemical parameters and a simple numerical coding, the liquid chromatography retention of bifunctionally substituted N-benzylideneaniles (NBA) has been predicted using a radial basis function neural network (RBFNN) model. The quantum chemical parameters involved in the model are dipole moment (m), energies of the highest occupied and lowest unoccupied molecular orbitals (E(homo,) E(lumo)), net charge of the most negative atom (Q(min)), sum of absolute values of the charges of all atoms in two given functional groups (Delta), total energy of the molecule (E(T)), weight of the molecule (W), and numerical coding (N). N was used to indicate the different positions of two substituents. The predictive values are consistent with the experimental results. The mean relative error of the testing set is 1.6%, and the maximum relative error is less than 5.0%. In this work the success of the whole modeling process only depends on the optimization of the spread parameter in network. PMID- 12086520 TI - The models of proton assisted and the unassisted formation of CGC base triplets. AB - The triple helix is formed by combining a double and a single strand DNAs in low pH and dissociates in high pH. Under such conditions, protonation of cytosine in the single strand is necessary for triplex formation where cytosine-guanine cytosine (CGC+) base triplet stabilizes the triple helix. The mechanism of CGC+ triplet formation from guanine-cytosine (GC) and a protonated cytosine (C+) shows the importance of N3 proton. Similarly in the case of CGC (unprotonated) triplet, the donor acceptor H-bond at N3 hydrogen of the cytosine analog (C) initiates the interaction with GC. The correspondence between the two models of triplets, CGC+ and CGC, unambiguously assigned that protonation at N3 cytosine in low pH to be the first step in triplet formation, but a donor acceptor triplet (CGC) can be designed without involving a proton in the Hoogsteen H-bond. Further, the bases of cytosine analogue also show the capability of forming Watson Crick (WC) H bonds with guanine. PMID- 12086521 TI - Effective descriptions of molecular structures and the quantitative structure activity relationship studies. AB - In this research, we found CoMFA alone could not obtain sufficiently a strong equation to allow confident prediction for aminobenzenes. When some other parameter, such as heat of molecular formation of the compounds, was introduced into the CoMFA model, the results were improved greatly. It gives us a hint that a better description for molecular structures will yield a better prediction model, and this hint challenged us to look for another method--the projection areas of molecules in 3D space for 3D-QSAR. It is surprising that much better results than that obtained by using CoMFA were achieved. Besides the CoMFA analysis, multiregression analysis and neural network methods for building the models were used in this paper. PMID- 12086522 TI - Evaluation of a novel electronic eigenvalue (EEVA) molecular descriptor for QSAR/QSPR studies: validation using a benchmark steroid data set. AB - A novel electronic eigenvalue (EEVA) descriptor of molecular structure for use in the derivation of predictive QSAR/QSPR models is described. Like other spectroscopic QSAR/QSPR descriptors, EEVA is also invariant as to the alignment of the structures concerned. Its performance was tested with respect to the CBG (corticosteroid binding globulin) affinity of 31 benchmark steroids. It appeared that the electronic structure of the steroids, i.e., the "spectra" derived from molecular orbital energies, is directly related to the CBG binding affinities. The predictive ability of EEVA is compared to other QSAR approaches, and its performance is discussed in the context of the Hammett equation. The good performance of EEVA is an indication of the essential quantum mechanical nature of QSAR. The EEVA method is a supplement to conventional 3D QSAR methods, which employ fields or surface properties derived from Coulombic and van der Waals interactions. PMID- 12086523 TI - Automatic spectra interpretation, structure generation, and ranking. AB - A new rule-based spectra interpretation system is described. It directly processes spectral files generated by spectrometers. Its internal information representation and the simultaneous use of several spectroscopic methods allow for the achievement of both high reliability and good performance. For small and medium sized molecules, the new program is capable of automatically reducing the solution space to under 3%. PMID- 12086524 TI - Basic charge clusters and predictions of membrane protein topology. AB - The topology predictor SPLIT 4.0 (http://pref.etfos.hr) predicts the sequence location of transmembrane helices by performing an automatic selection of optimal amino acid attribute and corresponding preference functions. The best topological model is selected by choosing the highest absolute bias parameter that combines the bias in basic charge motifs and the bias in positive residues (the "positive inside rule") with the charge difference across the first transmembrane segment. Basic charge motifs, such as the BBB, BXXBB, and BBXXB motifs in alpha-helical integral membrane proteins, are significantly more frequent near cytoplasmic membrane surface than expected from the Arg/Lys (B) frequency. The predictor's accuracy is 99% for predicting 178 transmembrane helices in all membrane proteins or subunits of known 3D structure. PMID- 12086525 TI - Determination of urinary calcium-oxalate formation risk with BONN-Risk-Index and EQUIL applied to a family. AB - Apart from environmental and acquired risk factors, a person's genetic predisposition may have a distinct influence on the probability of the onset of urolithiasis. To investigate the family related development of calcium oxalate, CaOx, crystallization risk, we studied urines from three generations of the same family. The paternal line has been suffering from CaOx-urolithiasis for at least two generations; no case of urolithiasis has been reported from the maternal line and the youngest generation.We applied the BONN-Risk-Index and the computer program EQUIL to determine the crystallization risk of each family member (n = 7). We clearly verified by probability calculations of the existence of the two risk groups within the family and showed that one of the siblings of the youngest generation may have inherited the stone-formation risk from its paternal relatives as this person clearly reflects a high risk pattern. PMID- 12086526 TI - Exploring the limits of graph invariant- and spectrum-based discrimination of (sub)structures. AB - The limits of a recently proposed computer method for finding all distinct substructures of a chemical structure are systematically explored within comprehensive graph samples which serve as supersets of the graphs corresponding to saturated hydrocarbons, both acyclic (up to n = 20) and (poly)cyclic (up to n = 10). Several pairs of smallest graphs and compounds are identified that cannot be distinguished using selected combinations of invariants such as combinations of Balaban's index J and graph matrix eigenvalues. As the most important result, it can now be stated that the computer program NIMSG, using J and distance eigenvalues, is safe within the domain of mono- through tetracyclic saturated hydrocarbon substructures up to n = 10 (oligocyclic decanes) and of all acyclic alkane substructures up to n = 19 (nonadecanes), i.e., it will not miss any of these substructures. For the regions surrounding this safe domain, upper limits are found for the numbers of substructures that may be lost in the worst case, and these are low. This taken together means that the computer program can be reasonably employed in chemistry whenever one is interested in finding the saturated hydrocarbon substructures. As to unsaturated and heteroatom containing substructures, there are reasons to conjecture that the method's resolving power for them is similar. PMID- 12086527 TI - Modeling blood-brain barrier partitioning using the electrotopological state. AB - The challenging problem of modeling blood-brain barrier partitioning is approached through topological representation of molecular structure. A QSAR model is developed for in vivo blood-brain partitioning data treated as the logarithm of the blood-brain concentration ratio. The model consists of three structure descriptors: the hydrogen E-State index for hydrogen bond donors, HS(T)(HBd); the hydrogen E-State index for aromatic CHs, HS(T)(arom); and the second order difference valence molecular connectivity index, d(2)chi(v) (q(2) = 0.62.) The model for the set of 106 compounds is validated through use of an external validation test set (20 compounds of the 106, MAE = 0.33, rms = 0.38), 5 fold cross-validation (MAE = 0.38, rms = 0.47), prediction of +/- values for an external test set (27/28 correct), and estimation of logBB values for a large data set of 20 039 drugs and drug-like compounds. Because no 3D structure information is used, computation of logBB by the model is very fast. The quality of the validation statistics supports the claim that the model may be used for estimation of logBB values for drug and drug-like molecules. Detailed structure interpretation is given for the structure indices in the model. The model indicates that molecules that penetrate the blood-brain barrier have large HS(T)(arom) values (presence of aromatic groups) but small values of HS(T)(HBd) (fewer or weaker H-Bond donors) and smaller d(2)chi(v) values (less branched molecules with fewer electronegative atoms). These three structure descriptors encode influence of molecular context of groups as well as counts of those groups. PMID- 12086528 TI - On molecular polarizability. 4. Evaluation of the ionization potential for alkanes and alkenes with polarizability. AB - To express the influence of polarizability effect on ionization potential (Ip), the Geometric Mean Polarizability Effect Index (GMPEI) and Geometric Mean Polarizability Effect Index of pi Bond (GMPEIpi) were proposed for alkanes and alkenes, respectively. Taking a few of the compounds as a model, we obtained the correlation equations between the experimental Ip and GMPEI or GMPEI pi and then, with the obtained equations, we evaluated the Ip for the title compounds. The estimated Ip values by this work are in good agreement with the measured ones. Furthermore, the ab initio (#HF/6-31G** OPT, Gaussian 98 program) and semiempirical (AM1) Ip computation are finished, and other topological index correlations with the Ip of alkane are discussed. PMID- 12086529 TI - Recent advances in PC-Linux systems for electronic structure computations by optimized compilers and numerical libraries. AB - One of the most frequently used packages for electronic structure research, GAUSSIAN 98, is compiled on Linux systems with various hardware configurations, including AMD Athlon (with the "Thunderbird" core), AthlonMP, and AthlonXP (with the "Palomino" core) systems as well as the Intel Pentium 4 (with the "Willamette" core) machines. The default PGI FORTRAN compiler (pgf77) and the Intel FORTRAN compiler (ifc) are respectively employed with different architectural optimization options to compile GAUSSIAN 98 and test the performance improvement. In addition to the BLAS library included in revision A.11 of this package, the Automatically Tuned Linear Algebra Software (ATLAS) library is linked against the binary executables to improve the performance. Various Hartree-Fock, density-functional theories, and the MP2 calculations are done for benchmarking purposes. It is found that the combination of ifc with ATLAS library gives the best performance for GAUSSIAN 98 on all of these PC-Linux computers, including AMD and Intel CPUs. Even on AMD systems, the Intel FORTRAN compiler invariably produces binaries with better performance than pgf77. The enhancement provided by the ATLAS library is more significant for post-Hartree Fock calculations. The performance on one single CPU is potentially as good as that on an Alpha 21264A workstation or an SGI supercomputer. The floating-point marks by SpecFP2000 have similar trends to the results of GAUSSIAN 98 package. PMID- 12086530 TI - Structure/response correlations and similarity/diversity analysis by GETAWAY descriptors. 1. Theory of the novel 3D molecular descriptors. AB - Novel molecular descriptors based on a leverage matrix similar to that defined in statistics and usually used for regression diagnostics are presented. This leverage matrix, called Molecular Influence Matrix (MIM), is here proposed as a new molecular representation easily calculated from the spatial coordinates of the molecule atoms in a chosen conformation. The proposed molecular descriptors are called GETAWAY (GEometry, Topology, and Atom-Weights AssemblY) as they try to match 3D-molecular geometry provided by the molecular influence matrix and atom relatedness by molecular topology, with chemical information by using different atomic weightings (atomic mass, polarizability, van der Waals volume, and electronegativity, together with unit weights). A first set of molecular descriptors, called H-GETAWAY, is derived by using only the information provided by the molecular influence matrix, while a second set, called R-GETAWAY, combines this information with geometric interatomic distances in the molecule. The prediction ability in structure-property correlations of the new descriptors was tested by analyzing regressions of these descriptors for selected properties of octanes. PMID- 12086531 TI - Structure/response correlations and similarity/diversity analysis by GETAWAY descriptors. 2. Application of the novel 3D molecular descriptors to QSAR/QSPR studies. AB - In a previous paper the theory of the new molecular descriptors called GETAWAY (GEometry, Topology, and Atom-Weights AssemblY) was explained. These descriptors have been proposed with the aim of matching 3D-molecular geometry, atom relatedness, and chemical information. In this paper prediction ability in structure-property correlations of GETAWAY descriptors has been tested extensively by analyzing the regressions of these descriptors for selected properties of some reference compound classes. Moreover, the general performance of the new descriptors in QSAR/QSPR has been evaluated with respect to other well known sets of molecular descriptors. PMID- 12086532 TI - Transition state structure and energetics of the N(2)O + X (X = Cl,Br) reactions. AB - The structural and vibrational properties of the transition state of the N(2)O + X (X = Cl,Br) reactions have been characterized by ab initio methods using density functional theory. We have employed Becke's hybrid functional (B3LYP), and transition state optimizations were performed with 6-31G(d), 6-311G(2d,2p), 6 311+G(3d,2p), and 6-311+G(3df,2p) basis sets. For the chlorine atom reaction the coupled-cluster method (CCSD(T)) with 6-31G(d) basis set was also used. All calculations resulted in transition state structures with a planar cis arrangement of atoms for both reactions. The geometrical parameters of transition states at B3LYP are very similar, and the reaction coordinates involve mainly the breaking of the N-O bond. At CCSD(T)/6-31G(d) level a contribution of the O-Cl forming bond is also observed in the reaction coordinate. In addition, several highly accurate ab initio composite methods of Gaussian-n (G1, G2, G3), their variations (G2(MP2), G3//B3LYP), and complete basis set (CBS-Q, CBS-Q//B3LYP) series of models were applied to compute reaction energetics. All model chemistries predict exothermic reactions. The G3 and G2 methods result in the smallest deviations from experiment, 1.8 and 0 kcal mol(-1), for the enthalpies of reaction for N(2)O reaction with chlorine and bromine, respectively. The G3//B3LYP and G1 methods perform best among the composite methods in predicting energies of the transition state, with a deviation of 1.9 and 3.0 kcal mol(-1), respectively, in the activation energies for the above processes. However, the B3LYP/6-311+G(3df,2p) method gives smaller deviations of 0.4 and -1.0 kcal mol( 1), respectively. The performance of the methodologies applied in predicting transition state energies was analyzed. PMID- 12086533 TI - A cellular automata model of water structuring by a chiral solute. AB - The organization of water around a solute molecule with surface features of varying hydropathic states is studied. A stationary solute molecule and mobile water solvent molecules are modeled using cellular automata dynamics. It is shown that varying hydropathic states of solute molecule surface features influence the relative affinities of water for these features. In the case of a simulated chiral solute, a chiral pattern of associated water molecules binding to the surface is produced. This finding is in agreement with published simulations and circular dichroism measurements. A pattern of water molecules at locations beyond the surface of the solute molecules is detected, evidence of an emergent property in this solvent-solute system. PMID- 12086534 TI - Neural network studies. 4. Introduction to associative neural networks. AB - Associative neural network (ASNN) represents a combination of an ensemble of feed forward neural networks and the k-nearest neighbor technique. This method uses the correlation between ensemble responses as a measure of distance amid the analyzed cases for the nearest neighbor technique. This provides an improved prediction by the bias correction of the neural network ensemble. An associative neural network has a memory that can coincide with the training set. If new data becomes available, the network further improves its predictive ability and provides a reasonable approximation of the unknown function without a need to retrain the neural network ensemble. This feature of the method dramatically improves its predictive ability over traditional neural networks and k-nearest neighbor techniques, as demonstrated using several artificial data sets and a program to predict lipophilicity of chemical compounds. Another important feature of ASNN is the possibility to interpret neural network results by analysis of correlations between data cases in the space of models. It is shown that analysis of such correlations makes it possible to provide "property-targeted" clustering of data. The possible applications and importance of ASNN in drug design and medicinal and combinatorial chemistry are discussed. The method is available on line at http://www.vcclab.org/lab/asnn. PMID- 12086535 TI - The construction and assessment of a statistical model for the prediction of protein assay data. AB - The focus of this work is the development of a statistical model for a bioinformatics database whose distinctive structure makes model assessment an interesting and challenging problem. The key components of the statistical methodology, including a fast approximation to the singular value decomposition and the use of adaptive spline modeling and tree-based methods, are described, and preliminary results are presented. These results are shown to compare favorably to selected results achieved using comparitive methods. An attempt to determine the predictive ability of the model through the use of cross-validation experiments is discussed. In conclusion a synopsis of the results of these experiments and their implications for the analysis of bioinformatic databases in general is presented. PMID- 12086536 TI - A marine natural product database. AB - A database of marine natural products has been developed. The database contains approximately 6000 chemical compounds derived from over 10,000 marine-derived materials. For each compound, the structure, physical and chemical properties, marine source, and biological activities are given. A computer program for searching this database has also been developed and is described. PMID- 12086538 TI - Accurate partitioning of compounds belonging to diverse activity classes. AB - Diverse sets of compounds were classified according to biological activity by use of a partitioning approach based on principal component analysis in conjunction with a genetic algorithm for molecular descriptor evaluation. Combinations of 236 molecular property and structural key descriptors were explored for their performance in classifying 317 molecules belonging to 21 distinct biological activity classes from various sources. Preferred descriptor combinations were further explored by complete factorial analysis. In these calculations, compounds having similar specific activity were predicted with greater than 80% accuracy. PMID- 12086537 TI - Combined MEDV-GA-MLR method for QSAR of three panels of steroids, dipeptides, and COX-2 inhibitors. AB - The MEDV-13, molecular electronegativity distance vector based on 13 atomic types, has at best 91 descriptors. It is impossible to indirectly use multiple linear regression (MLR) to derive a quantitative structure-activity relationship (QSAR) model. Although principal component regression (PCR) or partial least squares regression (PLSR) can be employed to develop a latent QSAR model, it is still difficult how to determine the principal components (PCs) and depict the physical meaning of the PCs. So, a genetic algorithm (GA) is first employed to select an optimal subset of the descriptors from original MEDV-13 descriptor set. Then MLR is utilized to build a QSAR model between the optimal subset and the biological activities of three sets of compounds. For 31 benchmark steroids, a 5 descriptor QSAR model (M1) between the corticosteroid-binding globulin (CBG) binding affinity of the steroids and 5-descriptor subset is developed. The root mean-square error of estimations (RMSEE) and the correlation coefficient of estimations (r) between the CBG binding affinity (BA) observed and the BA estimated by M1 are 0.422 and 0.9182, respectively. The root-mean-square error of predictions (RMSEP) and the correlation coefficient of predictions (q) between the BA observed and the BA predicted by leave-one-out cross validations are 0.504 and 0.8818, respectively. For 58 dipeptides inhibiting angiotensin-converting enzyme (ACE), a 5-variable QSAR model (M2) between the pIC(50) of peptides and 5 descriptor subset is derived. The M2 has a high quality with RMSEE = 0.339 and r = 0.9398 and RMSEP = 0.370 and q = 0.9280. For 16 indomethacin amides and esters (ImAE) inhibiting cyclooxygenase-2 (COX-2), a 6-variable QSAR model (M3) with RMSEE = 0.079 and r = 0.9839 and RMSEP = 0.151 and q = 0.9413 is built. PMID- 12086539 TI - The Chemistry Preprint Server: an experiment in scientific communication. PMID- 12086540 TI - Chemistry preprints. PMID- 12086542 TI - Neurologic, psychological, and aggressive disturbances with sildenafil. AB - BACKGROUND: Anecdotal reports and case studies have described psychological disturbances and aggressive behavior in some men taking sildenafil. In the course of assisting the defense in a trial in which a man was alleged to have committed rape and in which sildenafil was implicated, a mechanistic hypothesis was formulated by the first author for central nervous system (CNS) adverse effects associated with the use of sildenafil. OBJECTIVE: To examine whether there was any scientific information available to suggest a possible mechanism for or an association between exposure to sildenafil and aggressive behavior. METHODS: The scientific literature on sildenafil, nitric oxide (NO), and the NO-cyclic guanosine monophosphate (cyclic-GMP) signaling pathway was reviewed. Adverse event reports that referenced sildenafil and were filed with the Food and Drug Administration Adverse Event Reporting System between January 4, 1998, and February 21, 2001, also were examined. RESULTS: Published studies reported that sildenafil crosses the blood-brain barrier, that it exerts various biochemical and physiologic effects in the brain, and that it affects information processing. Other published reports indicated that phosphodiesterase type 5 (PDE-5), NO synthase, and guanylyl cyclase are present at highest activities in areas of the brain responsible for behavior, sexual drive, and emotion, and that NO modulates aggression and sexual behavior in male mice. In addition, 274 adverse event reports designated sildenafil as the primary suspect of various neurologic disturbances, amnesia, and aggressive behavior. DISCUSSION: Evidence has been presented for an association between sildenafil and various CNS adverse effects, including aggressive behavior. Whether sildenafil causes these effects by inhibiting PDE-5 in the brain, accumulating cyclic-GMP, decreasing NO, and affecting cell-cell signaling and modulation of aggressive behavior requires further investigation. CONCLUSIONS: It is recommended that, before prescribing sildenafil for erectile dysfunction, clinicians should caution their patients and their partners on the possibility of neurologic, emotional, or psychological disturbances; amnesia or loss of consciousness; or aggressive behavior. PMID- 12086543 TI - Cost impact of switching histamine(2)-receptor antagonists to nonprescription status. AB - BACKGROUND: There is a recent trend to switching medications from prescription to nonprescription status. Often, such switches are accompanied by dramatic changes in utilization due to increased availability or decreased insurance coverage. The histamine(2)-receptor antagonists (H(2)RAs) underwent such status change in the UK in 1994, the US in 1995, and Canada in 1996. OBJECTIVE: To examine the impact of the status change for H(2)RAs on the market for gastrointestinal (GI) agents in the US, UK, and Canada. METHODS: IMS market sales data from 1992 to 1997 were procured. All costs were converted to 1997 US dollars using the consumer price index. Per capita sales figures were determined using population data from the US Census Bureau's International Database. RESULTS: Overall spending on GI remedies increased in all 3 markets between 1992 and 1997; however, the contribution of prescription sales and number of prescriptions varied across the 3 countries. An increased market share for nonprescription H(2)RAs occurred in the US, correlating with a decline in prescription numbers for GI remedies. The opposing trend occurred in the UK, where market share of nonprescription H(2)RAs was minimal and use of prescription H(2)RAs increased. Prescription and nonprescription H(2)RA sales could not be differentiated for Canada. CONCLUSIONS: The impact of the H(2)RA status change varied across countries. Differences in utilization may be attributed to many factors such as differing healthcare systems, patient convenience, and physician prescribing practices. Further research is required to identify the reasons for differences in utilization and to quantify the potential clinical impact. PMID- 12086544 TI - 24-hour ambulatory blood pressure monitoring in male children receiving stimulant therapy. AB - OBJECTIVE: To determine whether cardiac indices are altered as assessed by 24 hour ambulatory blood pressure monitoring (ABPM) in male children receiving either chronic methylphenidate or dextroamphetamine/levoamphetamine (Adderall) therapy. METHODS: Boys 7-11 years old who were receiving methylphenidate or Adderall for a minimum of 2 months were asked to participate. Subjects wore ambulatory blood pressure monitors for 24-hour periods both off and on stimulant therapy. RESULTS: Subjects (n = 17; 8 methylphenidate, 9 Adderall) were well matched. Systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate differed between off and on stimulant therapy (p < 0.05). DBP load calculated from ABPM reference data was increased significantly (9.0% +/- 5.6% on and 4.8% +/- 4.5% off therapy; p < 0.05) while subjects were taking Adderall. There was a trend toward a greater elevation in blood pressure load during awake hours and a more pronounced decrease during the asleep hours for periods on compared with off-stimulant therapy. This trend resulted in significant (p < 0.05) nocturnal dipping on-stimulant phases compared with off-stimulant therapy for both SBP and DBP (Adderall) and SBP (methylphenidate). Two subjects (1 Adderall, 1 methylphenidate) met the criteria to be considered hypertensive based both on mean awake and 24-hour blood pressure load assessments during their on treatment period. One additional subject receiving Adderall therapy met the criteria to be considered hypertensive based on blood pressure load criteria while off therapy only. Positive correlation coefficients (p < 0.05) were found when comparing stimulant dose (mg/kg) with the percent change of mean SBP, DBP, and heart rate between off and on therapy (r = 0.56, 0.61, and 0.58, respectively). CONCLUSIONS: These preliminary data suggest that blood pressure and heart rate appear to be altered in male patients while receiving stimulant therapy for attention-deficit hyperactivity disorder. Blood pressure and heart rate screening and monitoring during stimulant therapy to determine whether alterations become clinically significant is encouraged. PMID- 12086545 TI - Safety of benzodiazepines in newborns. AB - BACKGROUND: Benzodiazepines are being used in neonatal intensive care units for sedation and control of seizures. However, anecdotal reports suggest that their use in infants may be associated with serious adverse effects (AEs). OBJECTIVE: To determine the incidence of AEs from benzodiazepine use in preterm and full term infants. METHODS: Retrospective chart review of 63 infants who received benzodiazepines as a sedative or anticonvulsant over a 16-month period. RESULTS: Mean +/- SD gestational age of the infants was 33.1 +/- 6.2 weeks, and birth weight was 2.3 +/- 1.2 kg. Median (range) postnatal age at commencement of drug administration was 19 (5-54) days. Forty-one infants received lorazepam, 8 received midazolam, and 14 received both. Ten (16%) of the infants had 14 documented adverse events: seizures (n = 6), hypotension (n = 5), and respiratory depression (n = 3). Using a validated adverse drug reaction probability scale, a probable association with benzodiazepine use was demonstrated in 12 of the AEs. Due to the retrospective nature of the data, a score for definite association was not attainable. Anticonvulsant administration was required for 4 of 6 infants and, in all cases of respiratory depression, ventilatory support was initiated or increased. Two cases of significant hypotension were treated with inotropes. There was no statistically significant correlation between AEs and benzodiazepine dose or concomitant use of inotropes or analgesics (morphine), although most infants had underlying medical conditions or received multiple drugs that may have predisposed them to experience AEs. CONCLUSIONS: Administration of benzodiazepines was frequently associated with AEs in full-term and preterm infants. It is possible that underlying illnesses and concomitant drug use predisposed these effects. Until the benefit-to-risk ratio is determined by further studies, judicious use of benzodiazepines is recommended in this vulnerable population. PMID- 12086546 TI - Serum quinidine concentrations and effect on QT dispersion and interval. AB - OBJECTIVE: To establish a relationship between serum quinidine concentrations (SQCs) and QT interval dispersion, compared with corresponding QT intervals, in order to identify a reason why many reports describe torsade de pointes as occurring at subtherapeutic concentrations. DESIGN: Retrospective study. SETTING: University teaching hospital. PARTICIPANTS: Eleven patients with atrial arrhythmias managed with quinidine therapy. MAIN OUTCOME MEASURES: Patients with subtherapeutic (<2 microg/mL) and therapeutic (2-5 microg/mL) SQCs with corresponding 12-lead electrocardiograms (ECGs) (25 mm/sec) and baseline ECG were evaluated for QT interval dispersion, calculated as the maximum minus the minimum QT interval on the 12-lead ECG. RESULTS: Mean +/- SD subtherapeutic and therapeutic SQCs were 1.48 +/- 0.39 microg/mL and 3.78 +/- 0.88 microg/mL (p < 0.001). Baseline values for QT/QTc intervals were 376.4 +/- 59.2/429.5 +/- 57.3 msec. At subtherapeutic and therapeutic SQCs, mean QT/QTc intervals were 403.6 +/ 59.9/450.5 +/- 38.5 msec and 439.1 +/- 48.9/472.4 +/- 44.6 msec, respectively. Mean QT dispersion was 47 +/- 16.2 msec at baseline, 98.2 +/- 27.5 msec at subtherapeutic SQC, and 70.9 +/- 33.9 msec at therapeutic SQCs (p = 0.001 for overall analysis; p < 0.001 for baseline vs. subtherapeutic concentrations; p = NS for therapeutic vs. subtherapeutic in post hoc comparison). CONCLUSIONS: Despite QT interval lengthening with increasing SQCs, QT dispersion was numerically greatest at subtherapeutic SQCs. Further study is required to determine the value of QT dispersion as a tool for identifying proarrhythmic risk with drugs that prolong the QT interval. PMID- 12086547 TI - Possible gatifloxacin-induced fulminant hepatic failure. AB - OBJECTIVE: To report a case of fulminant hepatic failure associated with the use of gatifloxacin. CASE SUMMARY: A 76-year-old white man was found to have an approximate 1-week history of worsening jaundice on the last day of a 10-day course of gatifloxacin for treatment of impetigo while at his skilled nursing facility. Liver function tests including aspartate aminotransferase (AST), alanine aminotransferase (ALT), international normalized ratio, activated partial thromboplastin time, and ammonia concentrations were found to be markedly elevated, consistent with hepatocellular necrosis commonly seen with fluoroquinolones. Screening for other causes of hepatotoxicity, including alcoholic or viral hepatitis, obstruction, or autoimmune-mediated processes, were negative. Other potential medication causes were less likely. The patient's liver function steadily declined, eventually resulting in multiple organ failure. The patient died 25 days after completing the course of gatifloxacin. DISCUSSION: This case of hepatotoxicity was associated with gatifloxacin. Other fluoroquinolones, most notably trovafloxacin, have been observed to cause variable degrees of hepatotoxicity, ranging from asymptomatic elevations of liver enzymes to fulminant hepatic failure. Fluoroquinolones are thought to cause hepatocellular necrosis, which results in elevated ALT and AST concentrations with a normal alkaline phosphatase concentration. A variable degree of hyperbilirubinemia is often seen, with the presence and degree of jaundice often correlating to a poorer prognosis. Hepatic encephalopathy and coagulopathy are also commonly present. CONCLUSIONS: Fluoroquinolones, including trovafloxacin, ciprofloxacin, ofloxacin, enoxacin, norfloxacin, and, in this case report, gatifloxacin, have been associated with hepatotoxicity. It is important that these medications be considered a possible cause when the patient being treated has liver disease. PMID- 12086548 TI - Constipation, polyuria, polydipsia, and edema associated with orlistat. AB - OBJECTIVE: To report the occurrence of a novel group of adverse effects associated with initiation and rechallenge of orlistat. CASE SUMMARY: A 42-year old white woman developed symptoms of constipation, polyuria, polydipsia, and increased lower-leg edema after 2 weeks of treatment with orlistat 120 mg 3 times daily. The drug was discontinued for 4 days and the symptoms resolved. On reinstitution of the orlistat treatment, the symptoms reappeared within 2 days. Thereafter, the medication was permanently discontinued. DISCUSSION: Common gastrointestinal adverse reactions associated with orlistat use include fecal urgency and abdominal pain and discomfort. Pedal edema has also been reported to occur, although less frequently. No reports were discovered documenting the occurrence of constipation, polydipsia, and polyuria associated with the use of orlistat. Despite careful consideration of other possible causes of these symptoms, the temporal association between initiation, discontinuation, and rechallenge of orlistat and the patient's symptoms suggest a medication-related adverse event. Based on the Naranjo probability scale, the likelihood that orlistat was the cause of this cluster of adverse effects is possible. CONCLUSIONS: It is important for the healthcare provider to be aware of these adverse effects to promptly evaluate and differentiate between possible causes of similar reactions. PMID- 12086549 TI - Toxic epidermal necrolysis associated with interleukin-2. AB - BACKGROUND: Metastatic renal cell carcinoma (MRCC) has been characteristically unresponsive to chemotherapy. In lieu of an effective regimen, interleukin-2 (IL 2) and interferon alfa are considered drugs of choice to treat this cancer. Subcutaneous IL-2 is safe and well tolerated, with a mortality rate <3%. OBJECTIVE: To report a case of cutaneous and hematologic toxicity in a patient treated with IL-2. CASE SUMMARY: A 67-year-old woman received radiotherapy and immunotherapy for cancer that had metastasized to the bone and lungs. IL-2 was part of the regimen. After 5 days of treatment with IL-2, the patient developed a hemorrhagic lesion that progressed to toxic epidermal necrolysis, as well as grade 4 pancytopenia. She died 10 days after treatment was begun. At the time of death, leukocytes were 0.3 x 10(3)/mm(3), platelets 10 x 10(3)/mm(3), and hemoglobin 6.8 mg/dL. DISCUSSION: Cutaneous IL-2 adverse effects are frequent, but generally mild and reversible. The adverse hematologic effects are usually transitory and pancytopenia is not frequent. The severity of cutaneous and hematologic toxicity experienced by our patient has rarely been reported. CONCLUSIONS: The use of IL-2 in bedridden patients with performance status >2 must be given on an individualized basis. If radiotherapy over extensive areas of the body is needed, the use of IL-2 must be postponed until radiotherapy is completed. PMID- 12086550 TI - Delayed recurrent SIADH associated with SSRIs. AB - BACKGROUND: Depression is a common problem in elderly patients and is frequently treated with selective serotonin-reuptake inhibitors (SSRIs). OBJECTIVE: To report a case of delayed recurrent hyponatremia after switching from one SSRI to another. CASE SUMMARY: An 87-year-old depressed woman began treatment with fluvoxamine. One week later, she was diagnosed with hyponatremia, most likely syndrome of inadequate antidiuretic hormone. Following discontinuation of fluvoxamine, the serum sodium concentration normalized. Later, she began treatment with paroxetine. Sixteen months after initiating paroxetine, she developed symptomatic recurrent hyponatremia. After paroxetine was discontinued, the sodium concentration normalized. DISCUSSION: In this case, unlike those previously reported, hyponatremia recurred 16 months after a different SSRI was initiated. The Naranjo probability scale indicates a probable relationship between recurrent hyponatremia and paroxetine. The mechanism of SSRI-induced hyponatremia is multifactorial. CONCLUSIONS: This case illustrates that replacement of one SSRI with another can cause delayed, recurrent hyponatremia in elderly patients. Plasma sodium concentrations must be monitored, not only in the first weeks of treatment, but throughout the full course. PMID- 12086551 TI - Possible zoophilia associated with dopaminergic therapy in Parkinson disease. AB - OBJECTIVE: To report a patient with Parkinson disease (PD) who developed zoophilia as a possible complication of dopaminergic therapy. CASE SUMMARY: A 74 year-old man with advanced PD, who had wearing-off motor fluctuations, with a marked disability during the off periods and severe peak-of-dose choreiform dyskinesias, developed hypersexuality with zoophilia 5 days after standard levodopa was substituted for controlled-release levodopa and the dose of bromocriptine was increased. The abnormal sexual behavior disappeared 2 days after the doses of standard levodopa and of bromocriptine were reduced. DISCUSSION: Hypersexuality is a known complication in PD patients undergoing dopaminergic therapy. However, the possible development of zoophilia due to these drugs, as was the case in our patient, is exceptional. CONCLUSIONS: Zoophilia should be considered as a possible behavioral complication of dopaminergic therapy in PD patients. PMID- 12086552 TI - SIADH and hyponatremia with theophylline. AB - OBJECTIVE: To report a case of possible theophylline-induced hyponatremia due to the syndrome of inappropriate antidiuretic hormone (SIADH). CASE SUMMARY: An 88 year-old man developed severe symptomatic hyponatremia (serum sodium 112 mEq/L) associated with inappropriate natriuresis (urinary sodium 58 mEq/L) temporally related to the initiation of theophylline. The patient fulfilled the criteria for the diagnosis of SIADH after all other causes of hyponatremia were excluded. Furthermore, no other drugs or conditions that could have evoked SIADH were found. DISCUSSION: Theophylline has rarely been associated with hyponatremia. A thiazide-like action of the drug on the stimulation of SIADH could be the underlying mechanism for SIADH. CONCLUSIONS: Theophylline should be considered as a possible cause of hyponatremia. PMID- 12086553 TI - Darbepoetin alfa: a novel erythropoiesis-stimulating protein. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, clinical efficacy, and safety of darbepoetin alfa. DATA SOURCES: Pertinent references were identified by a MEDLINE search (1995-January 2001) of the medical literature, review of English language literature and references of these articles, product information, and abstracts from professional meetings. STUDY SELECTION: Clinical efficacy data were gathered from all available trial data citing darbepoetin alfa. Additional information concerning pharmacology, pharmacokinetics, and safety was also reviewed. DATA SYNTHESIS: Darbepoetin alfa is a new erythropoiesis-stimulating protein with a threefold longer half-life than recombinant human erythropoietin (r-HuEPO). Darbepoetin alfa is approved for intravenous and subcutaneous administration in patients requiring and not requiring dialysis. Clinical studies in patients with chronic kidney disease (CKD) have shown darbepoetin alfa to be equivalent to r-HuEPO in terms of increases in hemoglobin concentration, percentage of patients obtaining target hemoglobin, and average time to reach target hemoglobin concentration. Trials are currently ongoing in patients receiving cancer chemotherapy. The adverse event profile appears to be similar between the 2 agents. CONCLUSIONS: The equivalent efficacy and safety profile, as well as the longer half-life, may make darbepoetin alfa an attractive alternative to r-HuEPO in patients with CKD. Since these patients need to receive r-HuEPO 1-3 times weekly at the expense of increased healthcare utilization to improve their hemoglobin, agents such as darbepoetin alfa, with longer durations of action, may reduce healthcare expenses. In addition, enhanced patient compliance may be realized with once-weekly or once every-other-week administration. PMID- 12086554 TI - Kaletra (lopinavir/ritonavir). AB - OBJECTIVE: To review the pharmacology, virology, pharmacokinetics, efficacy, safety, and clinical use of lopinavir/ritonavir (Kaletra, Abbott Laboratories). DATA SOURCES: English-language MEDLINE and AIDSline searches were performed (1966 July 2001) using lopinavir, ABT-378, and Kaletra as key words. Abstracts from infectious diseases and HIV scientific meetings were identified. Abbott Laboratories provided additional published and unpublished information. DATA EXTRACTION: All publications, meeting abstracts, and unpublished information were reviewed and relevant items included. In vitro and preclinical studies were included as well as Phase II and III clinical trials. DATA SYNTHESIS: Lopinavir/ritonavir is a fixed-dose protease inhibitor (PI) combination used for the treatment of HIV-1 infection. Lopinavir, the active component of this combination, is extensively metabolized by CYP3A4 and produces low systemic concentrations when used alone. Ritonavir potently inhibits CYP3A4 and is used to enhance the systemic exposure of lopinavir. This combination results in lopinavir concentrations that greatly exceed those necessary in vitro to inhibit both wild type and PI-resistant HIV isolates. In clinical trials with antiretroviral naive and experienced patients, lopinavir/ritonavir was effective at suppressing HIV RNA and increasing CD4+ T cell counts. Compared with other PIs, lopinavir/ritonavir may have advantages in the areas of pharmacokinetics, efficacy, and resistance. Toxicity, drug interactions, and medication adherence are important considerations surrounding its clinical use. CONCLUSIONS: Lopinavir/ritonavir is an effective option for the treatment of HIV-1-infected individuals when used in combination with other antiretroviral agents. It may be used as a component of initial therapy or salvage therapy; future studies will better define its place in therapy. PMID- 12086556 TI - Orlistat use in type 2 diabetes. AB - OBJECTIVE: To review the use of orlistat in type 2 diabetes. DATA SOURCES: A MEDLINE search of the English-language literature (1990-August 2001) was performed using the key terms orlistat, obesity, glucose, and diabetes. DATA EXTRACTION: All articles pertaining to orlistat were considered for inclusion, with emphasis placed on randomized, placebo-controlled, double-blind clinical trials. DATA SYNTHESIS: In April 1999, orlistat was approved by the Food and Drug Administration for the treatment of obesity. Of 13 randomized, placebo-controlled studies, only 2 reported specific data in individuals with type 2 diabetes. Both reported significant weight reduction and improved glycemic control over placebo. CONCLUSIONS: Since weight loss is a difficult goal to achieve in patients with type 2 diabetes, orlistat can be a safe, effective addition to a multidisciplinary approach. PMID- 12086555 TI - Anakinra treatment of patients with rheumatoid arthritis. AB - OBJECTIVE: To summarize the safety data arising from clinical trials of anakinra, a human recombinant form of interleukin-1 receptor antagonist (IL-1Ra) developed for the treatment of rheumatoid arthritis (RA). DATA SOURCES: Primary articles and abstracts identified through the National Library of Medicine's PubMed database (1982-2001) and secondary sources. STUDY SELECTION AND DATA EXTRACTION: All the articles and abstracts identified from the data sources were evaluated and all information deemed relevant was included in this review. DATA SYNTHESIS: Overall, anakinra was shown in 4 trials to be efficacious as monotherapy or combination therapy, compared with placebo, for the treatment of active RA. The magnitude of clinical improvement tended to increase with increasing doses of anakinra. In monotherapy trials, the primary adverse event was injection-site reactions, which usually were mild and transient. In com bination therapy (anakinra plus methotrexate), the frequency and severity of adverse events were similar to those seen in the monotherapy trials. CONCLUSIONS: The use of anakinra in patients with RA inhibits disease activity with a favorable tolerability profile. PMID- 12086557 TI - Immunization recommendations for adults with cancer. AB - OBJECTIVE: To review the published literature on immunizing nonbone marrow transplant adult cancer patients, summarize the findings, and make recommendations for the use of vaccines in this population. DATA SOURCE: A search of MEDLINE and CancerLit was conducted (1966-June 2001) to find English-language clinical studies and review articles pertaining to immunization, vaccines, and cancer in humans. Recommendations of the Advisory Committee on Immunization Practices were used extensively. References of each identified article were subsequently reviewed for additional relevant articles. STUDY SELECTION AND DATA EXTRACTION: Representative epidemiologic reports, clinical trials, and recommendations of expert panels are summarized in this report. Relevant information was selected to describe the epidemiology of vaccine-preventable diseases, efficacy of the vaccines, and recommendations specific to adults with cancer. DATA SYNTHESIS: In general, adults with cancer are at least at the same risk of infection with vaccine-preventable diseases as are healthy populations. Because of their compromised immune function, many patients who have undergone cancer treatment are specifically at increased risk of morbidity and mortality associated with measles and varicella infections. Asplenic patients with lymphoma are at increased risk of fulminant bacterial infections. Influenza infection is associated with significant morbidity in cancer patients. Although the protection conferred by immunization is lower in immunosuppressed patients with cancer, immunization with inactivated vaccines is indicated. Live vaccines should not be used except in very rare instances. CONCLUSIONS: Immunization of adults with cancer is a critical component of their care. Although additional research is necessary, following established recommendations may protect individuals with malignancies from significant morbidity and mortality associated with vaccine preventable diseases. PMID- 12086558 TI - A critical evaluation of the therapeutic range of indinavir. AB - OBJECTIVE: To provide a summary of the patient population, study characteristics, and important findings of the key studies in the literature evaluating concentration-effect relationships and the therapeutic range for indinavir. DATA SOURCES: Literature search strategy involved using MEDLINE (1966-July 2001) and AIDSLINE (MEDSCAPE) databases (up to July 2001). Reference lists from primary literature and review articles were also examined, and conference abstracts were obtained. STUDY SELECTION: English-language articles were considered suitable for review if the clinical trials in HIV patients reported on concentration-effect relationships and/or pharmacokinetic breakpoints or threshold concentrations. A search of the literature identified 20 peer-reviewed references from 18 separate studies including journal articles and conference abstracts. DATA EXTRACTION: The targeted pharmacokinetic parameters and breakpoint values, the rationale for their selection or method of identification, and other study details and limitations were summarized. DISCUSSION: This article highlights the heterogeneity of studies evaluating the therapeutic range of indinavir and provides a summary of important findings of the key studies in the literature evaluating concentration-effect relationships and therapeutic range. Tables are provided to enable clinicians to make use of currently available information on the therapeutic range of indinavir. CONCLUSIONS: There is insufficient evidence to recommend a general therapeutic range for indinavir. Future investigations should incorporate both pharmacokinetics and pharmacodynamics in order to define a broadly applicable therapeutic range. PMID- 12086559 TI - Preventable drug-related hospital admissions. AB - OBJECTIVE: To estimate the prevalence of preventable drug-related hospital admissions (PDRAs) and to explore if selected study characteristics affect prevalence estimates. METHODS: Keyword search of MEDLINE (1966-December 1999), International Pharmaceutical Abstracts (1970-December 1999), and hand search. Two reviewers independently selected studies published in peer-reviewed journals and extracted crude prevalence estimates and study characteristics. Trials had to specifically address consequences of drug therapy requiring hospital admission and include a quantitative preventability assessment. Stratified analysis and meta-regression were used to explore the association between study characteristics and prevalence estimates. DATA SYNTHESIS: Fifteen studies reported a median PDRA prevalence of 4.3% (interquartile range [IQR] 3.1-9.5%). The median preventability rate of drug-related admissions was 59% (IQR 50-73%). No evidence of publication bias related to study size could be determined. Because the individual study results were highly heterogeneous (Cochran's Q = 176, df = 14; p < 0.001), no meta-analytic summary estimate was computed. Stratified analysis suggested an association between prevalence estimates and 3 study characteristics: exclusion of first admissions (readmission studies: average PDRA prevalence of 14.0 %, estimated prevalence OR = 3.7); mean age of admissions >70 (OR = 2.1); and inclusion of "indirect" drug-related morbidity, such as omission errors or therapeutic failure (OR = 1.9). There was little evidence of other associations with prevalence estimates, such as selection of specific hospital units, exclusion/inclusion of planned admissions, country, and specified methods of PDRA case ascertainment. CONCLUSIONS: Drug-related morbidity is a significant healthcare problem, and a great proportion is preventable. Study methods in prevalence reports vary and should be considered when interpreting findings or planning future research. PMID- 12086560 TI - Management of acute, severe asthma in children. AB - OBJECTIVE: To briefly present the current options available for the treatment of acute, severe asthma in children, with a special focus on emergency department and inpatient treatment, and to describe newer therapies that may aid treatment in the future. DATA SOURCES AND STUDY SELECTION: A MEDLINE search (1966-May 2001) of the English-language literature pertaining to drug therapy of acute asthma was performed. Key word searches included acute asthma, albuterol, ipratropium, corticosteroids, magnesium, and theophylline. Additional articles from these sources and published national guidelines were identified. Relevant studies pertaining to current therapy of acute asthma in pediatric patients were selected; if there were minimal pediatric data, adult data were included. DATA SYNTHESIS: Asthma is a chronic, inflammatory disorder of the airways. Acute exacerbations can occur and are challenging to manage. Albuterol, ipratropium, and systemic corticosteroids have been shown to be effective in acute asthma exacerbations. Because some patients do not respond to maximal therapy, older therapies such as magnesium and theophylline are being reevaluated. Theophylline may have some therapeutic effect, but given its toxicity profile, it is unclear whether it offers any advantage over maximal beta(2)-agonist therapy. There are only minimal published data evaluating the use of magnesium in pediatrics, and most are small trials or case reports. Newer therapies such as ventilation strategies with heliox and intravenous leukotriene modifiers currently being evaluated may or may not prove to be beneficial in the future. CONCLUSIONS: beta(2)-agonists, ipratropium, and corticosteroids remain the most useful therapeutic agents for acute asthma exacerbations in pediatric patients. However, these agents are not ideal in all patients and, given the existing questions regarding safety and/or efficacy of available alternatives, more effective options are needed. PMID- 12086561 TI - Acute renal failure in hospitalized patients: part I. AB - BACKGROUND: Acute renal failure (ARF) is a common condition in hospitalized patients. Morbidity, mortality, and health resource use are considerable, but the true magnitude of the problem is not well described in the literature. OBJECTIVE: To provide a detailed discussion of the epidemiology, economic costs, and classification of ARF. DATA SOURCES: A MEDLINE search (1996-December 2001) was conducted using the search terms kidney and acute kidney failure: epidemiology, etiology, and drug therapy/drug effects. Bibliographies of selected articles were also examined to include all relevant investigations. Economic data were identified using the terms costs and cost analysis and cost of illness. STUDY SELECTION AND DATA EXTRACTION: Review articles, meta-analyses, and clinical trials describing epidemiology and classification of hospital-acquired ARF were identified. Results from prospective, controlled trials were given priority when available. CONCLUSIONS: ARF occurs in up to 25% of critically ill patients, resulting in significant morbidity and high mortality. Characterization of ARF is difficult due to multiple etiologic factors and variable definitions. Limited cost data describe the extensive economic burden associated with the disorder, although further pharmacoeconomic research is needed. Epidemiology and classification of ARF allow prospective management of at-risk patients. PMID- 12086562 TI - Treatment of vitamin b(12)-deficiency anemia: oral versus parenteral therapy. AB - OBJECTIVE: To evaluate the use of oral cyanocobalamin therapy in the treatment of cobalamin (vitamin B(12))-deficient anemia. DATA SOURCES: Primary and review articles were identified by MEDLINE search (1966-May 2000) and through secondary sources. DATA SYNTHESIS: Cobalamin-deficient anemia is among the most common diagnoses in older populations. Cobalamin-deficient anemia may be diagnosed as pernicious anemia, resulting from the lack of intrinsic factor required for cobalamin absorption or as protein malabsorption from the inability to displace cobalamin from protein food sources. Several studies provide evidence that daily oral cyanocobalamin as opposed to monthly parenteral formulations may adequately treat both types of cobalamin-deficient anemias. CONCLUSIONS: Daily oral cyanocobalamin at doses of 1000-2000 microg can be used for treatment in most cobalamin-deficient patients who can tolerate oral supplementation. There are inadequate data at the present time to support the use of oral cyanocobalamin replacement in patients with severe neurologic involvement. PMID- 12086563 TI - Estrogen replacement therapy for Alzheimer disease in postmenopausal women. AB - OBJECTIVE: To evaluate the clinical utility of oral conjugated equine estrogen in postmenopausal women with Alzheimer disease. DATA SOURCES: Literature was identified through MEDLINE (1997-January 2002). Key search terms included Alzheimer disease, estrogen replacement therapy, and treatment. DATA SYNTHESIS: Estrogen has been identified as a neuroprotective agent with possible application in degenerative disorders. Observational studies have demonstrated an association between estrogen replacement therapy and decreased incidence of Alzheimer disease. Two recent, controlled clinical trials have evaluated the role of oral conjugated estrogen in the treatment of Alzheimer disease. CONCLUSIONS: Clinical trials indicate that oral conjugated equine estrogen is not an effective treatment for Alzheimer disease in postmenopausal women. PMID- 12086564 TI - The use of topiramate in mood stabilization. AB - OBJECTIVE: To review data on the effectiveness of topiramate as a mood stabilizer. DATA SOURCES: Clinical literature accessed through MEDLINE (1985 September 2001) and the manufacturer. Key search terms included topiramate, mania, mood stabilizer, and bipolar disorder. DATA SYNTHESIS: The traditional standard therapy for bipolar disorder has been lithium. Other mood stabilizers are increasingly being used to manage this complex disorder. Studies that used topiramate in bipolar disorders were evaluated. CONCLUSIONS: The present data from open trials suggest that topiramate may possibly possess antimanic properties. Controlled, double-blind studies are required to confirm this efficacy. PMID- 12086565 TI - Celecoxib-associated torsade de pointes. PMID- 12086566 TI - Anagrelide-induced erectile dysfunction. PMID- 12086567 TI - Comment: risk of extrapyramidal syndromes with haloperidol, risperidone, or olanzapine. PMID- 12086569 TI - Comment: neuroleptic malignant syndrome associated with risperidone and fluvoxamine. PMID- 12086571 TI - Comment: secretin for autism: unproven treatment or ineffective treatment? PMID- 12086573 TI - Comment: severe pancytopenia associated with methotrexate therapy for rheumatoid arthritis. PMID- 12086574 TI - Perspectives on rehabilitation of individuals with cognitive impairment after brain injury: rationale for reconsideration of theoretical paradigms. AB - BACKGROUND: The frequency of cognitive impairment associated with traumatic brain injury (TBI) has led to the widespread use of cognitive rehabilitation as a discrete rehabilitative service. This service has become controversial in part because of disagreements regarding its theoretical base and implementation procedures, and in part because of insufficient experimental support. OBJECTIVE: We outline two paradigms for cognitive rehabilitation that are interestingly different, both theoretically and procedurally. These paradigms are defined in relation to the goals of intervention, orientation to assessment, modalities of treatment, organization of treatment, and logistics of service delivery (setting, content, and provider). A rationale for each paradigm is described. CONCLUSION: We conclude with a call for (a) additional research into the effectiveness of interventions for individuals with cognitive impairment after TBI and (b) clinicians' thoughtful reflection on the foundations of their service. PMID- 12086575 TI - Measuring recovery of orientation during acute rehabilitation for traumatic brain injury: value and expectations of recovery. AB - OBJECTIVES: To further evaluate the use of the orientation log (O-Log) in an acute rehabilitation program for adults with traumatic brain injury (TBI), specifically focusing on prediction of rehabilitation outcome, possible use of the O-Log to promote recovery, and development of templates of orientation recovery. DESIGN: Stepwise discriminant function analysis was used in one study and ANOVA based on between-groups comparisons in another. Confidence intervals were established in the third study. SETTING: Acute rehabilitation hospital. PATIENTS: Samples were taken from an overall pool of 389 subjects with predominantly severe TBI based on Glasgow Coma Scale score. RESULTS: A combination of initial O-Log performance, time since injury, and number of O-Log assessments correctly predicted resolution of disorientation for 76% of the sample. Individuals attaining orientation before discharge achieved higher discharge scores on functional measures. Administration of the O-Log 3 versus 5 times a week did not result in more rapid resolution of disorientation. A similar pattern of steady improvement in orientation was evident for mild-moderate and severe injury cases across 10 administrations. CONCLUSIONS: Recovery of orientation provides important information about recovery following TBI and is predictive of rehabilitation outcome. More frequent administration of a formal orientation measure does not seem to expedite the recovery of orientation, which on average seems to progress at a steady pace in a rehabilitation setting regardless of the time since injury or severity of injury. PMID- 12086576 TI - Acute characteristics of pediatric Dysphagia subsequent to traumatic brain injury: videofluoroscopic assessment. AB - OBJECTIVE: To document the acute characteristics of swallowing impairment in a group of children post moderate/severe traumatic brain injury (TBI) by means of videofluoroscopy. PARTICIPANTS: Eighteen children with moderate/severe TBI. MAIN OUTCOME MEASURE: Videofluoroscopy at an average of 27.7 days post-injury. RESULTS: Subjects demonstrated a range of dysphagia severity levels: mild moderate (n = 8), moderate (n = 6), moderate-severe (n = 3), and severe (n = 1) and had a combination of oral and pharyngeal phase characteristics. More specifically, observable features or physiological impairments that were identified included reduced lingual control, hesitancy of tongue movement, repetitive tongue pumping, the presence of aspiration (including silent aspiration), delayed swallow reflex trigger, reduced laryngeal elevation and closure, and reduced peristalsis. CONCLUSIONS: These data highlight the diversity of swallowing deficits and dysphagia severity levels in children following TBI and suggest that the former are consistent with a pattern of oropharyngeal impairments. PMID- 12086577 TI - Telerehabilitation needs: a survey of persons with acquired brain injury. AB - OBJECTIVE: To survey individuals with acquired brain injury to assess multiple facets of interest, access, and familiarity necessary to implement new telerehabilitation technologies. DESIGN: Anonymous mail survey. SETTING: Community. PARTICIPANTS: Seventy-one respondents to a survey. These individuals had experienced acquired brain injury (predominantly severe traumatic brain injury [TBI]) and were living in the community. Surveys were mailed by a state chapter of the Brain Injury Association to a random selection of members with acquired brain injury. MAIN OUTCOME MEASURE: Survey designed specifically for this investigation. RESULTS: The survey responses indicate that there is great interest in the possibility of accessing telerehabilitative services among individuals with acquired brain injury. In particular, there was strong interest expressed in services that could be used to assist with problems in memory, attention, problem-solving, and activities of daily living. CONCLUSIONS: Telemedicine, and more specifically telerehabilitation, holds great promise as an adjunct to traditional clinical service delivery. Little research in this area has been applied, however, to individuals with acquired brain injuries. Although on the surface, telerehabilitation seems to be an appropriate assessment and treatment modality for individuals with brain injury, it will only succeed if those individuals have the interest-and the access-necessary to use new and evolving technologies. PMID- 12086578 TI - Assessment of anosmia after traumatic brain injury: performance characteristics of the University of Pennsylvania Smell Identification Test. AB - OBJECTIVE: To examine the performance characteristics of two forms of the University of Pennsylvania Smell Identification Test (UPSIT) in a sample of persons with traumatic brain injury (TBI). DESIGN: Analysis of consecutive admissions into a brain injury rehabilitation program. SETTING: Midwestern medical center. PARTICIPANTS: One hundred twenty-two adults diagnosed with TBI (49% severe TBI, 16% moderate TBI, 35% mild TBI). MAIN OUTCOME MEASURES: University of Pennsylvania Smell Identification Test (UPSIT; 3- and 40-item versions). RESULTS: Fifty-six percent of sample exhibited impaired olfaction on the full UPSIT; 40% of these patients were unaware of their deficits. Contrary to expectation, TBI patients detected dangerous odors (natural gas, gasoline, smoke) with high accuracy. Usefulness of a 3-item screening measure was examined: Missing even one item related to a 2:1 likelihood of being anosmic. CONCLUSIONS: These findings support past investigations indicating that anosmia, and unawareness of olfactory dysfunction, are common in persons with TBI and related to injury severity. The use of the 3-item screening measure as a gross indicator was supported, although caution is advised, because nearly 20% of patients performing perfectly on the 3-item screen scored in the anosmic range on the full UPSIT. PMID- 12086579 TI - Morbidity and management conference: an approach to quality improvement in brain injury rehabilitation. AB - BACKGROUND: Program Evaluation and Quality Improvement (QI) are an ongoing focus of all brain injury rehabilitation programs, but it is difficult to consistently include the entire rehabilitation team in QI activities and to focus on issues that cross the continuum of care. In addition, recent reports have provided a renewed emphasis on reduction of medical errors and improving the overall quality of care for individuals with chronic conditions. Rehabilitation Medicine has a long history of working as a coordinated multidisciplinary team caring for complex patients with chronic conditions and is in a position to be a leader in QI activities. DESIGN: We have modified the traditional Morbidity and Mortality conference to include a multidisciplinary case discussion to identify system issues with quality of care. SETTING: This is completed in a nonpunitive format that does not assign individual blame. CONCLUSION: This Rehabilitation Morbidity and Management Conference is an important addition to the QI activities of our program. PMID- 12086580 TI - A novel, kinetically stable, catalytically active, all-ferric, nitrite-bound complex of Paracoccus pantotrophus cytochrome cd1. AB - The oxidized form of Paracoccus pantotrophus cytochrome cd(1) nitrite reductase, as isolated, has bis-histidinyl co-ordination of the c haem and His/Tyr co ordination of the d(1) haem. On reduction, the haem co-ordinations change to His/Met and His/vacant respectively. If the latter form of the enzyme is reoxidized, a conformer is generated in which the ferric c haem is His/Met co ordinated; this can revert to the 'as isolated' state of the enzyme over approx. 20 min at room temperature. However, addition of nitrite to the enzyme after a cycle of reduction and reoxidation produces a kinetically stable, all-ferric complex with nitrite bound to the d(1) haem and His/Met co-ordination of the c haem. This complex is catalytically active with the physiological electron donor protein pseudoazurin. The effective dissociation constant for nitrite is 2 mM. Evidence is presented that d(1) haem is optimized to bind nitrite, as opposed to other anions that are commonly good ligands to ferric haem. The all-ferric nitrite bound state of the enzyme could not be generated stoichiometrically by mixing nitrite with the 'as isolated' conformer of cytochrome cd(1) without redox cycling. PMID- 12086582 TI - The functional consequences of mis-sense mutations affecting an intra-molecular salt bridge in arylsulphatase A. AB - Metachromatic leukodystrophy is a lysosomal storage disorder caused by the deficiency of arylsulphatase A. We describe the functional consequences of three mis-sense mutations in the arylsulphatase A gene (Asp-335-Val, Arg-370-Trp and Arg-370-Gln), affecting an apparent intramolecular Asp-335 to Arg-370 salt bridge, and interpret the effects and clinical consequences on the basis of the three-dimensional structure of arylsulphatase A. Asp-335-Val and Arg-370-Trp substitutions each cause a complete loss of enzyme activity and are associated with the most severe form of the human disease, whereas the Arg-370-Gln substituted enzyme retains some residual activity, being found in a patient suffering from the milder juvenile form of the disease. Detailed analysis reveals that formation of the apparent salt bridge depends critically on the presence of aspartic acid and arginine residues at positions 335 and 370, respectively. Substitution by various other amino acids, including glutamic acid and lysine, affects enzyme function severely. Biosynthesis and immunoprecipitation studies indicate that the Asp-335-Val substitution affects folding of arylsulphatase A more severely than either the Arg-370-Trp or Arg-370-Gln substitutions. In vitro mutagenesis data show that clinical severity correlates with the space occupied by residue 370. The combination with structural data suggests that the bulky tryptophan residue broadens the cleft held together by the apparent salt bridge, whereas the smaller glutamine residue still allows the cleft to close, yielding a less severely affected enzyme. The position of residue 370 in the three dimensional structure of the enzyme provides a plausible explanation for the differing severities in loss of enzyme function caused by the mutations and thus the clinical phenotype. PMID- 12086583 TI - Cathepsin B expression and down-regulation by gene silencing and antisense DNA in human chondrocytes. AB - Cathepsin B, a marker of the dedifferentiated chondrocyte phenotype, contributes to cartilage destruction in osteoarthritis and pathological proteolysis in rheumatoid arthritis and cancer. In search of possible means for neutralizing the action of this enzyme, we compared its expression, biosynthesis and distribution in articular chondrocytes and two lines of immortalized human chondrocytes. Native articular chondrocytes in primary culture and the polyclonal T/C-28a2 chondrocyte cell line were similar with respect to the number of endosomes and lysosomes, the distribution of three alternatively spliced cathepsin B mRNA forms, and the cathepsin B activity. In contrast, the clonal C-28/I2 cell line contained four times higher levels of intracellular cathepsin B activity, slightly higher numbers of endosomes and lysosomes, and uniform distribution of all three cathepsin B transcripts and thus resembled subcultured chondrocytes at an early stage of dedifferentiation. Transfection of T/C-28a2 chondrocytes with double-stranded cathepsin B mRNA resulted in inhibition of cathepsin B biosynthesis by up to 70% due to RNA interference, and single-stranded antisense DNAs of various sizes decreased cathepsin B biosynthesis by up to 78%. An antisense oligonucleotide designed to hybridize to the end of cathepsin B's exons 1 and the beginning of exon 3 was successful in specifically inhibiting the mRNA splice variant lacking exon 2. These results indicate that cathepsin B expression and activity may be targeted for gene silencing by RNA interference and antisense DNA in chondrocytes. Furthermore, the differential expression and distribution of cathepsin B and presence of the necessary molecular apparatus for gene silencing in the immortalized human chondrocyte cell lines indicate that they may serve as a useful model for studying the function of relevant enzymes in cartilage pathologies. PMID- 12086581 TI - Identification of a novel AU-rich-element-binding protein which is related to AUF1. AB - The AU-rich element (ARE) is an important instability determinant for a large number of early-response-gene mRNAs. AREs also mediate the stabilization of certain pro-inflammatory mRNAs, such as tumour necrosis factor (TNF)-alpha and cyclo-oxygenase-2 (COX-2), in response to inflammatory stimuli. To understand how AREs control mRNA stability, it is necessary to identify trans-acting factors. We have purified a new ARE-binding protein and identified it as CArG box-binding factor-A (CBF-A). The amino acid sequence of CBF-A is highly similar to that of the ARE-binding protein AUF1. Recombinant CBF-A bound the COX-2 and TNF-alpha AREs, but not a non-specific control RNA. In contrast, in an electrophoretic mobility-shift assay (EMSA) of crude RAW 264.7 macrophage-like cell extracts, an antiserum that recognizes both AUF1 and CBF-A failed to supershift complexes formed on the TNF-alpha ARE, but did supershift a complex specific for the COX-2 ARE. CBF-A exists as two isoforms, p37 and p42, that differ by a 47-amino-acid insertion close to the C-terminus. By expressing epitope-tagged isoforms of CBF-A it was shown that the p42 isoform binds the COX-2 ARE in EMSA of crude cell extracts. In a HeLa-cell tetracycline-regulated reporter system, overexpression of the p42 CBF-A isoform resulted in stabilization of a COX-2 ARE reporter mRNA. Epitope-tagged p42 CBF-A expressed in HeLa cells co-immunoprecipitated with endogenous COX-2 mRNA, but not glyceraldehyde-3-phosphate dehydrogenase mRNA, as shown by reverse-transcription PCR. The similarity between CBF-A and AUF1 suggests that CBF-A could be re-named AUF2. PMID- 12086584 TI - Induction of apoptosis in human leukaemic cells by IPENSpm, a novel polyamine analogue and anti-metabolite. AB - Human promyelogenous leukaemic cells (HL-60) were treated with novel spermine analogue, ( S )- N (1)-(2-methyl-1-butyl)- N (11)-ethyl-4,8-diazaundecane (IPENSpm), and the effects on growth and intracellular polyamine metabolism were measured. IPENSpm was cytotoxic to these cells at concentrations greater than 2.5 microM. It induced apoptosis in a caspase-dependent manner and its toxicity profile was comparable with etoposide, a well-known anti-tumour agent and inducer of apoptosis. IPENSpm decreased intracellular polyamine content as a result of changes in ornithine decarboxylase activity and increases in spermidine/spermine N(1)-acetyltransferase and polyamine export. Analysis showed spermine and spermidine as the major intracellular polyamines, while putrescine and acetyl polyamines were the main export compounds. IPENSpm used the polyamine transporter system for uptake and its accumulation in cells was prevented by polyamine transport inhibitors. IPENSpm can be classified as a polyamine anti-metabolite and it may be a promising new lead compound in terms of treatment of some human cancers. PMID- 12086585 TI - Insertion of a small peptide of six amino acids into the beta7-beta8 loop of the p51 subunit of HIV-1 reverse transcriptase perturbs the heterodimer and affects its activities. AB - BACKGROUND: HIV-1 RT is a heterodimeric enzyme, comprising of the p66 and p51 subunits. Earlier, we have shown that the beta7-beta8 loop of p51 is a key structural element for RT dimerization (Pandey et al., Biochemistry 40: 9505, 2001). Deletion or alanine substitution of four amino acid residues of this loop in the p51 subunit severely impaired DNA binding and catalytic activities of the enzyme. To further examine the role of this loop in HIV-1 RT, we have increased its size such that the six amino acids loop sequences are repeated in tandem and examined its impact on the dimerization process and catalytic function of the enzyme. RESULTS: The polymerase and the RNase H activities of HIV-1 RT carrying insertion in the beta7-beta8 loop of both the subunits (p66INS/p51INS) were severely impaired with substantial loss of DNA binding ability. These enzymatic activities were restored when the mutant p66INS subunit was dimerized with the wild type p51. Glycerol gradient sedimentation analysis revealed that the mutant p51INS subunit was unable to form stable dimer either with the wild type p66 or mutant p66INS. Furthermore, the p66INS/p66INS mutant sedimented as a monomeric species, suggesting its inability to form stable homodimer. CONCLUSION: The data presented herein indicates that any perturbation in the beta7-beta8 loop of the p51 subunit of HIV-1 RT affects the dimerization process resulting in substantial loss of DNA binding ability and catalytic function of the enzyme. PMID- 12086586 TI - Aged garlic extract therapy for sickle cell anemia patients. AB - BACKGROUND: Sickle cell anemia is one of the most prevalent hereditary disorders with prominent morbidity and mortality. With this disorder oxidative, phenomena play a significant role in its pathophysiology. One of the garlic (Allium sativum L.) formulations, aged garlic extract (AGE), has been reported to exert an anti oxidant effect in vitro, we have evaluated the anti-oxidant effect of AGE on sickle red blood cells (RBC). METHODS: Five patients (two men and three women, mean age 40 ± 15 years, range 24-58 years) with sickle cell anemia participated in the study. AGE was administered at a dose of 5 ml a day. Whole blood samples were obtained at baseline and at 4 weeks for primarily Heinz body analysis. RESULTS: The data were consistent with our hypothesis. In all patients, the number of Heinz bodies decreased over the 4 week period (58.9 ± 20.0% at baseline to 29.8 ± 15.3% at follow-up, p = 0.03). CONCLUSIONS: These data suggest that there is a significant anti-oxidant activity of AGE on sickle RBC. AGE may be further evaluated as a potential therapeutic agent to ameliorate complications of sickle cell anemia. PMID- 12086587 TI - Bouveret's syndrome complicated by distal gallstone ileus after laser lithotropsy using Holmium: YAG laser. AB - BACKGROUND: Bouveret's syndrome is an unusual presentation of duodenal obstruction caused by the passage of a large gallstone through a cholecystoduodenal fistula. Endoscopic therapy has been used as first-line treatment, especially in patients with high surgical risk. CASE PRESENTATION: We report a 67-year-old woman who underwent an endoscopic attempt to fragment and retrieve a duodenal stone using a Holmium: Yttrium-Aluminum-Garnet Laser (Ho:YAG) which resulted in small bowel obstruction. The patient successfully underwent enterolithotomy without cholecystectomy or closure of the fistula. CONCLUSION: We conclude that, distal gallstone obstruction, due to migration of partially fragmented stones, can occur as a possible complication of laser lithotripsy treatment of Bouveret's syndrome and might require urgent enterolithotomy. PMID- 12086588 TI - Early recovery of microvascular perfusion induced by t-PA in combination with abciximab or eptifibatide during postischemic reperfusion. AB - BACKGROUND: GPIIb/IIIa inhibitors abciximab and eptifibatide have been shown to inhibit platelet aggregation in ischemic heart disease. Our aim was to test the efficacy of abiciximab (Reo Pro) or eptifibatide (Integrilin) alone or in combination with plasminogen activator (t-PA) in an experimental model of ischemia reperfusion (I/R) in hamster cheek pouch microcirculation visualized by fluorescence microscopy. Hamsters were treated with saline, or abiciximab or eptifibatide or these drugs combined with t-PA infused intravenously 10 minutes before ischemia and through reperfusion. We measured the microvessel diameter changes, the arteriolar red blood cell (RBC) velocity, the increase in permeability, the perfused capillary length (PCL), and the platelet and leukocyte adhesion on microvessels. RESULTS: I/R elicited large increases in the platelet and leukocyte adhesion and a decrease in microvascular perfusion. These responses were significantly attenuated by abiciximab or eptifibatide (PCL:70 and 65% at 5 10 mins of reperfusion and 85 and 87% at 30 mins of reperfusion, respectively, p < 0.001) while t-PA combined with abiciximab or eptifibatide, was more effective and microvascular perfusion recovered immediately after postischemic reperfusion. CONCLUSIONS: Platelets are crucial in I/R injury, as shown by the treatment with abicixmab or eptifibatide, which decreased platelet aggregation in microvessels, and also decreased leukocyte adhesion in venules. Arterial vasoconstriction, decreased arterial RBC velocity and alterations in the endothelial barrier with increased permeability delayed the complete restoration of blood flow, while t-PA combined with inhibition of platelet aggregation speeded up the capillary perfusion after reperfusion. PMID- 12086589 TI - A new way to rapidly create functional, fluorescent fusion proteins: random insertion of GFP with an in vitro transposition reaction. AB - BACKGROUND: The jellyfish green fluorescent protein (GFP) can be inserted into the middle of another protein to produce a functional, fluorescent fusion protein. Finding permissive sites for insertion, however, can be difficult. Here we describe a transposon-based approach for rapidly creating libraries of GFP fusion proteins. RESULTS: We tested our approach on the glutamate receptor subunit, GluR1, and the G protein subunit, alphas. All of the in-frame GFP insertions produced a fluorescent protein, consistent with the idea that GFP will fold and form a fluorophore when inserted into virtually any domain of another protein. Some of the proteins retained their signaling function, and the random nature of the transposition process revealed permissive sites for insertion that would not have been predicted on the basis of structural or functional models of how that protein works. CONCLUSION: This technique should greatly speed the discovery of functional fusion proteins, genetically encodable sensors, and optimized fluorescence resonance energy transfer pairs. PMID- 12086592 TI - Modelling developmental instability as the joint action of noise and stability: a Bayesian approach. AB - BACKGROUND: Fluctuating asymmetry is assumed to measure individual and population level developmental stability. The latter may in turn show an association with stress, which can be observed through asymmetry-stress correlations. However, the recent literature does not support an ubiquitous relationship. Very little is known why some studies show relatively strong associations while others completely fail to find such a correlation. We propose a new Bayesian statistical framework to examine these associations RESULTS: We are considering developmental stability - i.e. the individual buffering capacity - as the biologically relevant trait and show that (i) little variation in developmental stability can explain observed variation in fluctuating asymmetry when the distribution of developmental stability is highly skewed, and (ii) that a previously developed tool (i.e. the hypothetical repeatability of fluctuating asymmetry) contains only limited information about variation in developmental stability, which stands in sharp contrast to the earlier established close association between the repeatability and developmental instability. CONCLUSION: We provide tools to generate valuable information about the distribution of between-individual variation in developmental stability. A simple linear transformation of a previous model lead to completely different conclusions. Thus, theoretical modelling of asymmetry and stability appears to be very sensitive to the scale of inference. More research is urgently needed to get better insights in the developmental mechanisms of noise and stability. In spite of the fact that the model is likely to represent an oversimplification of reality, the accumulation of new insights could be incorporated in the Bayesian statistical approach to obtain more reliable estimation. PMID- 12086590 TI - USF2 inhibits C/EBP-mediated transcriptional regulation of the RIIbeta subunit of cAMP-dependent protein kinase. AB - BACKGROUND: Cyclic AMP-dependent protein kinase (PKA) plays a central role in regulation of energy metabolism. Upon stimulation of testicular Sertoli cells by follicle stimulating hormone (FSH), glycolysis is activated to increase the production of nutrients for the germ cells, and a new regulatory subunit of cAMP dependent protein kinase, RIIbeta, is induced. We have previously shown that production of the transcription factor C/EBPbeta is rapidly increased by FSH and cAMP in primary Sertoli cell cultures, and that C/EBPbeta induces the RIIbeta promoter. RESULTS: In this work we show that USF1, USF2 and truncated USF isoforms bind to a conserved E-box in the RIIbeta gene. Interestingly, overexpression of USF2, but not USF1, led to inhibition of both cAMP- and C/EBPbeta-mediated induction of RIIbeta. Furthermore, Western blots show that a novel USF1 isoform is induced by cAMP in Sertoli cells. CONCLUSIONS: These results indicate that the expression of various USF isoforms may be regulated by cAMP, and that the interplay between USF and C/EBPbeta is important for cAMP mediated regulation of RIIbeta expression. The counteracting effects of USF2 and C/EBPbeta observed on the RIIbeta promoter is in accordance with the hypothesis that C/EBP and USF play opposite roles in regulation of glucose metabolism. PMID- 12086591 TI - Obtaining reliable information from minute amounts of RNA using cDNA microarrays. AB - BACKGROUND: High density cDNA microarray technology provides a powerful tool to survey the activity of thousands of genes in normal and diseased cells, which helps us both to understand the molecular basis of the disease and to identify potential targets for therapeutic intervention. The promise of this technology has been hampered by the large amount of biological material required for the experiments (more than 50 microg of total RNA per array). We have modified an amplification procedure that requires only 1 microg of total RNA. Analyses of the results showed that most genes that were detected as expressed or differentially expressed using the regular protocol were also detected using the amplification protocol. In addition, many genes that were undetected or weakly detected using the regular protocol were clearly detected using the amplification protocol. We have carried out a series of confirmation studies by northern blotting, western blotting, and immunohistochemistry assays. RESULTS: Our results showed that most of the new information revealed by the amplification protocol represents real gene activity in the cells. CONCLUSION: We have confirmed a powerful and consistent cDNA microarray procedure that can be used to study minute amounts of biological tissue. PMID- 12086593 TI - Representation of women's health in general medical versus women's health specialty journals: a content analysis. AB - BACKGROUND: Women's health, traditionally defined, emphasises reproductive and maternal conditions without consideration of social contexts. Advocates urge a broader conceptualisation. The medical literature influences the definitions and delivery of women's health care. We compared how women's health was represented in leading general medical (GM) versus women's health specialty (WS) journals. METHODS: Original investigations published between January 1 - June 30, 1999 in leading GM (n = 514) and WS (n = 82) journals were compared. Data were collected from 99 GM and 82 WS articles on women's health. Independent reviewers conducted content analyses of sample characteristics, study design, and health topic. Each article was classified as "Traditional" (e.g. menstruation, breast cancer), "Non traditional" (e.g. abuse, osteoporosis), or "Both." RESULTS: Of the GM articles, 53 (53.5%) focused solely on a traditional women's health topic; half were reproductive and half female cancers. In contrast, 22 (26.8%) WS articles were traditionally focused. A non-traditional topic was the sole focus of 27 (27.3%) GM articles versus 34 (41.5%) WS articles. One-fifth of GM and one-third of WS articles addressed both. RCTs dominated the GM articles, while 40% of WS articles used qualitative or mixed study designs. Leading sources of women's death and disability were not well covered in either type of journal. CONCLUSIONS: Most GM articles drew on a narrow definition of women's health. WS journals provided more balanced coverage, addressing social concerns in addition to "navel-to-knees" women's health. Since GM journals have wide impact, editorial decisions and peer review processes should promote a broader conceptualisation of women's health. PMID- 12086594 TI - Prescribing patterns in premenstrual syndrome. AB - BACKGROUND: Over 300 therapies have been proposed for premenstrual syndrome. To date there has been only one survey conducted in the UK of PMS treatments prescribed by GPs, a questionnaire-based study by the National Association of Premenstrual Syndrome in 1989. Since then, selective serotonin re-uptake inhibitors have been licensed for severe PMS/PMDD, and governmental recommendations to reduce the dosage of vitamin B6 (the first choice over-the counter treatment for many women with PMS) have been made. This study investigates the annual rates of diagnoses and prescribing patterns for premenstrual syndrome (1993-1998) within a computerised general practitioner database. METHODS: Retrospective survey of prescribing data for premenstrual syndrome between 1993-1998 using the General Practice Research Database for the West Midlands Region which contains information on 282,600 female patients RESULTS: Overall the proportion of women with a prescription-linked diagnosis of premenstrual syndrome has halved over the five years. Progestogens including progesterone were the most commonly recorded treatment for premenstrual syndrome during the whole study period accounting for over 40% of all prescriptions. Selective serotonin-reuptake inhibitors accounted for only 2% of the prescriptions in 1993 but rose to over 16% by 1998, becoming the second most commonly recorded treatment. Vitamin B6 accounted for 22% of the prescriptions in 1993 but dropped markedly between 1997 and 1998 to 11%. CONCLUSIONS: This study shows a yearly decrease in the number of prescriptions linked to diagnoses for premenstrual syndrome. Progestogens including progesterone, is the most widely prescribed treatment for premenstrual syndrome despite the lack of evidence demonstrating their efficacy. PMID- 12086595 TI - Regional differences in expression of VEGF mRNA in rat gastrocnemius following 1 hr exercise or electrical stimulation. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) mRNA levels increase in rat skeletal muscle after a single bout of acute exercise. We assessed regional differences in VEGF165 mRNA levels in rat gastrocnemius muscle using in situ hybridization after inducing upregulation of VEGF by treadmill running (1 hr) or electrical stimulation (1 hr). Muscle functional regions were defined as oxidative (primarily oxidative fibers, I and IIa), or glycolytic (entirely IIb or IId/x fibers). Functional regions were visualized on muscle cross sections that were matched in series to slides processed through in situ hybridization with a VEGF165 probe. A greater upregulation in oxidative regions was hypothesized. RESULTS: Total muscle VEGF mRNA (via Northern blot) was upregulated 3.5-fold with both exercise and with electrical stimulation (P = 0.015). Quantitative densitometry of the VEGF mRNA signal via in situ hybridization reveals significant regional differences (P PF-HPMC 2%>PF-CMC 2.5%>PF127 15%. The slowest drug release was obtained from 15% PF127 formulations containing 3% methylcellulose. In vivo study showed that the ocular bioavailability of TM, measured in albino rabbits, increased by 2.5 and 2.4 fold for 25% PF127 gel formulation and 15% PF127 containing 3% methylcellulose, respectively, compared with 0.5% TM aqueous solution. PMID- 12086721 TI - Plasma protein binding, lipoprotein distribution and uptake of free and lipid associated BCL-2 antisense oligodeoxynucleotides (G3139) in human melanoma cells. AB - The objectives of this study were to determine the protein binding and lipoprotein distribution of G3139 and G3139 lipoplexes following incubation in human plasma, assess complement activation of, and the effect of pre-incubation of G3139 and G3139 lipoplexes in human plasma on in vitro cellular uptake of G3139. Effect of concentration and time on incorporation of free and lipid associated (lipoplexes) [3H]Bcl-2 AO (25-600 ng/ml) into normolipidemic human plasma lipoproteins was determined by density gradient ultracentrifugation after incubation at 37 degrees C for 5, 30, 60 and 120 min. Protein binding in the lipoprotein deficient fractions (LPDP) was determined by equilibrium dialysis. Complement interaction was determined by ELISA after exposure of human plasma to AO+/- liposomes prepared in serial dilution. In vitro uptake of G3139 and G3139 lipoplexes into human melanoma cells was assessed qualitatively by fluorescence microscopy after 4-h exposure to G3139 (free or as lipoplexes) with or without pre-incubation of G3139 in normal human plasma. Analysis of Bcl-2 AO-lipoprotein interaction over time and concentration indicated no significant movement of the compound within the different lipoprotein and LPDP fractions. Majority of drug was recovered within LPDP fraction, and more than 85% of drug recovered within LPDP fraction was protein bound. No significant activation of complement was noted for either free AO or lipoplexes. Pre-incubation of free AO or AO lipoplexes in human plasma resulted in a greater cellular uptake of AO-lipoplexes compared with plasma free controls. These findings suggest that the majority of [3H]Bcl-2 AO is plasma protein bound with little lipoprotein association and no significant movement between different lipoprotein and LPDP fractions. Plasma protein binding other than lipoprotein binding may be responsible for the difference in cellular uptake of free AO vs. cationic lipoplexes. PMID- 12086722 TI - Failure of stability prediction for minodronic acid injectable by accelerated stability testing. AB - A liquid formulation containing 0.5 mg/ml minodronic acid, 40 mM, pH 4.5, citrate, and sodium chloride added to adjust the osmolarity of the final formulation was stored in flint glass ampoules at 25, 40, 50, and 60 degrees C. At specified times, the drug potency and pH, and the tendency to generate particulate matter, were measured. Test samples stored at 40 degrees C for 6 months or at 50 and 60 degrees C for 3 months were stable with no potency loss and no particulate increase. However, despite the satisfactory stability at high temperatures, the amount of particulate matter increased when the formulation was stored at 25 degrees C. Scanning electron microscopy-energy dispersive X-ray analysis of the particulate matter revealed that it contains aluminum and phosphorus, the latter thought to be derived from minodronic acid. In contrast, the number of the particulate matter did not increase, when the formulation was stored in either plastic containers or in SiO(2)-treated glass ampoules(.) The spike of minodronic acid solution with aluminum ions led to the particulate generation. These results demonstrate that the particulate matter is a complex of minodronic acid molecules and aluminum ions, which apparently leached from the glass of regular ampoules. Since the particulate generation could not be observed at higher temperatures, it was suggested that the complex formation was exothermic and accelerated testing did not predict the stability in terms of particulate generation. PMID- 12086723 TI - Comparison of the mechanical properties of the crystalline and amorphous forms of a drug substance. AB - PURPOSE: To better understand the influence of long-range molecular order on the processing characteristics of an active pharmaceutical ingredient (API). METHODS: Crystalline and amorphous samples of a model drug substance were isolated and their "true" density, crystallinity, melting point, glass transition temperature, particle size distribution, and powder flow characteristics determined. Compacts of a standard porosity were manufactured from each form and their dynamic indentation hardness, quasi-static indentation hardness, tensile strength and "compromised tensile strength" determined. X-ray powder diffraction was used to confirm that no changes were induced by compact formation or testing. RESULTS: The crystalline and amorphous forms of the drug substance had relatively high melting and glass transition temperatures (approximately 271 and 142 degrees C, respectively) and were physically and chemically stable under the conditions of the testing laboratory. Consistent with this there was no evidence of crystallinity in the amorphous samples or vice versa before, during or after testing. The two API lots were effectively equivalent in their particulate properties (e.g. particle size distribution), although differences in their particle morphologies were observed which influenced powder flow behavior. The compacts of the bulk drug samples exhibited moderate ductility, elasticity, and strength, and high brittleness, in keeping with many other drug substance samples. A significantly greater compression stress was required to form the compacts of the crystalline material, and these sample materials were more ductile, less brittle and less elastic than those made from the amorphous API. There were no major differences in the tensile strength or the viscoelasticity of the compacts made from the crystalline and amorphous samples. CONCLUSIONS: The mechanical properties of compacted amorphous and crystalline samples of a drug substance have been measured and the contributions due to the molecular ordering of the crystalline form proposed. Small but significant differences in the mechanical properties were noted which could potentially affect the processing performance of API. PMID- 12086724 TI - Development of level A, B and C in vitro-in vivo correlations for modified release levosimendan capsules. AB - The aim of this study was to investigate the possibility of developing different levels of correlation between in vitro release and in vivo absorption rate for four modified-release levosimendan capsule formulations. Differences and similarities in the in vitro dissolution curves were compared with pharmacokinetic parameters describing absorption rate. Formulations F, G, H and I differed in the amounts of the delaying excipients alginic acid and HPMC. In vitro release rate was studied by the USP basket method using the following conditions: pH 5.8 or 7.4 and a rotation speed of 50 or 100 rpm. In vivo bioavailability was tested in nine healthy male volunteers and the fractions absorbed were calculated by the Wagner-Nelson method. Dissolution conditions pH 5.8 and a rotation speed of 100 rpm predicted best the similarities and differences in absorption rates among different formulations, and levels C and B correlation coefficients were 0.85 and 0.97, respectively. For formulation H level A correlation (r=0.997) was found when in vitro lag time was 0.2 h and time scale factor 1.9. This study indicated that dissolution tests developed can be used as a surrogate for human bioequivalence studies, for development processes of final commercial products, to ensure batch to batch bioequivalence and in the future in possible scale-up and post approval change cases for modified-release levosimendan formulation H. PMID- 12086725 TI - The effect of ethanol on acrylic bone cement. AB - Prosthesis loosening is a major problem associated with the use of poly(methyl methacrylate) (PMMA) bone cement that may be related to a peri-implant vacuolisation commonly observed at bone-cement interface. Methyl methacrylate (MMA) monomer may be one of the cement components partly responsible for the mentioned vacuolisation due to a cytotoxic effect associated to this compound. Alcoholism has been related to bone necrosis in predisposed individuals. Furthermore, ethanol has been shown to clean material with adherent cement debris during cleaning procedure in laboratory. Consequently, we have decided to study whether ethanol will also be related to an increased liberation of MMA from the polymer matrix. 'In vitro' release studies using PMMA plates were conducted to access the role of ethanol on the liberation of the monomer. Contact angle measurements and surface tension estimation were also carried out in order to find a possible effect of ethanol on surface cement properties. Results suggest that ethanol, even in small quantities, enhances the leaching of the monomer from the polymer matrix, but does not considerably change the wettability properties of the cement surface. PMID- 12086727 TI - Evaluation of a new controlled-drug delivery concept based on the use of thermoresponsive polymers. AB - The purpose of this work is to develop a new delivery concept making a thermosensitive polymer based on poly(N-isopropylacrylamide) (PNIPAAm) useful as a time-controlled drug release device, without any temperature changes of the dissolution medium. It was previously found that some salts induce a decrease of the polymer lower critical solution temperature (LCST). Use is here made of that property to show that salt concentration variations can be used as a substitute for temperature changes to make the polymer coating of compression-coated tablets soluble or insoluble, consequently creating a possible new concept of drug delivery control from delivery systems containing thermoresponsive polymers. The obtained results show the influence of the type and amount of salts incorporated into compression-coated tablets on the release lag time of a model drug. PMID- 12086726 TI - Stabilization of amorphous indomethacin by co-grinding in a ternary mixture. AB - Mechanochemical amorphization of indomethacin (IM) was substantially enhanced by grinding with SiO(2), talc and a Mg(OH)(2)-SiO(2) mixture. The rates of the mechanochemical amorphization were in the order of Mg(OH)(2)-SiO(2) mixture>talc>SiO(2). Amorphous state stability of IM compounded with the carrier was examined by crystallization behavior under the condition of 30 degrees C and 11% relative humidity. Superiority of the binary mixture as a carrier was explained in terms of the mechanically induced strong acidic sites of the carrier. PMID- 12086728 TI - Buccal transport of flecainide and sotalol: effect of a bile salt and ionization state. AB - Patients with infrequent attacks of supraventricular arrhythmia may benefit from self administration of antiarrhythmic drugs on an 'as required' basis. The oral cavity is easily accessible and the potential for rapid absorption exists. The effects of ionization state and sodium glycocholate on the ex vivo transport of sotalol and flecainide across porcine buccal mucosa were studied. The permeated amounts at 3 h (Q) and fluxes (J) of sotalol in an aqueous solution at pH 7.4 and 9.0 were similar. At pH 7.4, in contrast to pH 9.0, the addition of 1.0% (w/v) sodium glycocholate decreased Q and J four and five fold. Flecainide base in propylene glycol resulted in a nine and 12 fold higher Q and J as compared with an aqueous solution of flecainide acetate at pH 5.8. The presence of sodium glycocholate reduced the transport rate of the flecainide base. However, Q and J were increased 110 and 75 fold by adding 1.0% (w/v) sodium glycocholate to a solution of flecainide acetate at pH 5.8. Sodium glycocholate seems to be an effective penetration enhancer for the buccal absorption of the more polar ionized form of flecainide in an aqueous solution. Sodium glycocholate does not seem to improve the transport of sotalol. PMID- 12086729 TI - Mucoadhesion on pig vesical mucosa: influence of polycarbophil/calcium interactions. AB - The influence of polycarbophil/calcium interactions on the mucoadhesive properties of polycarbophil has been examined. Polycarbophil dispersions and films with different concentrations of calcium or sodium ions were prepared and the following parameters were measured: detachment force on pig vesical mucosa, zeta potential, pH and viscosity. Polycarbophil detachment force decreased significantly in the presence of calcium but not sodium. Both ions decrease the pH of polycarbophil dispersions. On the other hand, altering the pH of hydrated polycarbophil films in the absence of added ions had an insignificant effect on detachment force. Both ions reduce the absolute values of polycarbophil zeta potential, calcium more efficiently than sodium. We could conclude that decreased mucoadhesion strength of polycarbophil in the presence of calcium is due to the chelation of polycarbophil carboxylic groups by calcium and crosslinking of polymer. The crosslinked polymer chains would be expected to be less flexible, and therefore, interpenetrate to a lesser extent with the glycosaminoglycans of mucus. Additionally, the interactions between functional groups of polycarbophil and mucus glycosaminoglycans are lowered due to the calcium, blocking the carboxylic groups. The mechanism of calcium influence on viscosity of polycarbophil dispersions appears to be different: repulsion between ionised carboxylic groups of polycarbophil prevails over the crosslinking of polycarbophil by calcium. PMID- 12086730 TI - Development of hemoglobin aquasomes from spherical hydroxyapatite cores precipitated in the presence of half-generation poly(amidoamine) dendrimer. AB - Spherical hydroxyapatite cores were prepared by using carboxylic acid terminated half-generation poly(amidoamine) (PAMAM) dendrimer as templates or crystal modifiers. The hydroxyapatite cores were characterized by infrared spectroscopy (IR), X-ray diffraction (XRD) and transmission electron microscopy (TEM). The spherical core formation depended on phosphate saturation, pH of the simulated body fluid (SBF) and rate of crystal growth. Hydroxyapatite so formed was amorphous and a mixture of various calcium phosphates. Ca/P ratio determination which showed phosphate rich apatite formation. Hydroxyapatite ores were coated with a sugar layer followed by hemoglobin to obtain aquasomes. Aquasomes were characterized for size, hemoglobin loading, oxygen-binding characteristics and storage stability. The nanometric sized aquasome formulation could load approximately 13.7 mg hemoglobin per g of core and retained oxygen-affinity and cooperativity and stability for at least 30 days. Formulation efficacy was tested in albino rats and indicated its potential utility as blood-substitute. PMID- 12086731 TI - Rheological evaluation of thermosensitive and mucoadhesive vaginal gels in physiological conditions. AB - The timely gelation and retention of in situ-gelling vaginal formulations would be fundamental to improve the efficacy of drugs. In this study, various rheological properties of clotrimazole gels were evaluated for predicting their performance in vagina. Two kinds of thermosensitive and mucoadhesive formulations were composed of poloxamer 407 (P407, 15%), polycarbophil (0.2%), and different amounts of P188 (15 vs. 20%). Both formulations were Newtonian at 20 degrees C but non-Newtonian at 37 degrees C. Although both liquid formulations gelled below the vaginal temperature, they differed in gelation time and viscoelastic properties in the presence of vaginal fluid simulant. At body temperature, the formulation with 20% of P188 gelled within 35 s but it took two times longer for the other one gelled. Upon dilution with simulated vaginal fluid, the formulation with 20% of P188 retained the rheology of a gel, but the other one lost the viscoelastic properties typical for a gel. Moreover, after dilution with simulated vaginal fluid, the elastic modulus was orders of magnitude higher in the formulations with 20% of P188 relative to the other one. These results indicate that the rheological evaluation at the physiologic conditions needs to be preceded to develop more effective in situ-gelling vaginal formulations. PMID- 12086732 TI - Bioavailability of a new ketoprofen formulation for once-daily oral administration. AB - A new sustained-release formulation (sustained release Ibifen) that gradually releases ketoprofen within 24 h and ensures therapeutic plasma concentration for the entire period has been developed. It consists of tableted pH-dependent barrier film-coated ketoprofen granules and was administered at a single dose of 200 mg to 12 volunteers. Ketoprofen plasma profiles were compared with: (1) administration of Orudis retard 200 capsule (200 mg); (2) two 12-h doses of prompt release Ibifen capsules (100 mg). In vitro dissolution kinetics and ketoprofen plasma levels were measured by HPLC. Sustained release Ibifen dissolution rate was constant for 10 h, whereas Orudis retard 200 dissolution profile presented one higher slope (0-6 h) and a lower one (6-12 h). Both formulations showed a delayed kinetics with respect to prompt release Ibifen. After sustained release Ibifen administration, ketoprofen plasma peak, reached within 2 h, remained practically constant for at least 12 h (average 4 microg/ml), which is higher than therapeutic levels. Differently, Orudis retard 200 produced a delayed, higher C(max) (5.91+/-0.66 vs. 4.51+/-0.65 microg/ml; P<0.01) and disappeared more quickly. In conclusion, sustained release Ibifen can ensure therapeutic ketoprofen plasma levels for the entire 24 h period, avoiding plasma concentration spikes, with bioavailability similar to other ketoprofen preparations. PMID- 12086733 TI - Influence of teardrop studs on rotating frictional base plate on spheroid quality in rotary spheronization. AB - The effects of teardrop-shaped studs on the quality of rotary processed spheroids were investigated. The spheroids were produced under similar conditions using three rotating frictional base plates with teardrop studs of different height, volume, cross-sectional area and surface area. Spheroid properties were rated by size, size distribution, shape, friability and density. The amounts of lumps and fines produced, and the adhesion of material on the rotating frictional base plates was also looked into. The dimension of the teardrop studs on the rotating frictional base plate affected spheroid quality. The resultant shear forces and energy input during rotary spheronization differed depending on the different height, volume, cross-sectional area and surface area of studs. With the increase in height, volume, cross-sectional area and surface area of studs on the frictional base plate, the mass median diameter, e(R) and circularity of spheroids increased with corresponding decrease in span, lumps and fines. Although the frictional base plate with shortest studs had little adhesion, it may not supply enough shear force and energy input for the spheronization process, resulting in a less stable process. A balance between energy input and adhesion on the rotating frictional base plate was needed in order to optimize the production of spheroids by rotary processing. PMID- 12086734 TI - In-vitro release of diclofenac diethylammonium from lipid-based formulations. AB - This article presents the preparation and topical performance of some new lipid based formulations of diclofenac, namely (a) a diclofenac aqueous gel containing mixed micelles (sodium cholate:egg lecithin molar ratio 0.55); (b) diclofenac lotion that contains soya lecithin, ethanol and buffer; and (c) diclofenac lipogel containing egg lecithin, isopropyl myristate, propylene glycol and ethanol. Gel formulations were prepared using Carbomer 934. Release of diclofenac from all formulations was monitored via dialysis through Spectra/por membrane into phosphate buffer (0.2 M pH=7.4) using a Franz cell. Drug release profile and diffusion coefficients were compared with brand formulation (Geigy's Vlotaren Emulgel). Statistical analysis of data show that the diffusion coefficient of the drug from these formulations rank according to the following order: Diclofenac lotion (D=5.308x10(-7) cm(2)/s) >lipogel (D=2.102 x 10(-7) cm(2)/s) >Voltaren Emulgel (1.518 x 10(-7) cm(2)/s) >aqueous gel mixed micelle (0.966 x 10(-7) cm(2)/s). These results show that diclofenac lotion and lipogel maybe more suitable formulations than the conventional topical dosage form. PMID- 12086735 TI - Giving drugs to knockout mice: can they do that? AB - The past ten years have seen an explosion in the number of genetically modified mice created to aid understanding of the basic mechanisms of learning and memory. There are still significant problems associated with this useful technique, mostly involving the lack of temporal or spatial control over the genetic 'lesion'. By combining the application of drugs that are sub-threshold in wild type mice with heterozygosity for gene mutations that do not produce effects alone, it is now possible to avoid many of the problems of both the genetic and the pharmacological approaches. PMID- 12086736 TI - The birth of a memory. AB - Laying down new memories has long been thought to involve interactions between the hippocampus and multiple regions of the neocortex. Functional neuroimaging studies performed over the past four years provide evidence for this proposal. A recent electrophysiological study offers a possible mechanism by which interactions between brain regions take place during memory formation. PMID- 12086738 TI - Competing on the edge. AB - Fast-acting neurotransmitters can exit the synaptic cleft and bind to extrasynaptic receptors. This process is modulated by transmitter uptake mechanisms (transporters). A new study focusing on glutamate-mediated transmission in the cerebellum describes the specific role of neuronal transporters in modulating the access of glutamate to extrasynaptic metabotropic glutamate receptors, and reveals important consequences of extrasynaptic signaling on synaptic plasticity. PMID- 12086740 TI - New laboratory start-up in the 21st century. AB - Launching a new neuroscience laboratory requires knowledge of purchasing, budgeting and personnel management. The usual training of scientific investigators omits these areas, at a time when experimental complexity is growing and Internet-based commerce offers new purchasing options. This review highlights laboratory start-up maxims and their applications to web-based purchasing, cost containment and budgeting. PMID- 12086739 TI - Jekyll-Hyde neurotrophins: the story of proNGF. AB - Neurotrophins promote neuronal survival and differentiation by binding to two classes of cell surface receptors: members of the Trk-family receptor tyrosine kinases and p75(NTR), a member of the tumor necrosis factor receptor superfamily. The discoveries that the unprocessed proneurotrophin peptide is both a preferential high-affinity ligand for p75(NTR) (with minimal affinity for trkA) and a potent inducer of p75(NTR)-mediated neuronal cell death are likely to change our understanding of neurotrophin physiology and our ability to manipulate their signaling pathways. PMID- 12086745 TI - Untangling spatial from temporal illusions. PMID- 12086747 TI - Exercise: a behavioral intervention to enhance brain health and plasticity. AB - Extensive research on humans suggests that exercise could have benefits for overall health and cognitive function, particularly in later life. Recent studies using animal models have been directed towards understanding the neurobiological bases of these benefits. It is now clear that voluntary exercise can increase levels of brain-derived neurotrophic factor (BDNF) and other growth factors, stimulate neurogenesis, increase resistance to brain insult and improve learning and mental performance. Recently, high-density oligonucleotide microarray analysis has demonstrated that, in addition to increasing levels of BDNF, exercise mobilizes gene expression profiles that would be predicted to benefit brain plasticity processes. Thus, exercise could provide a simple means to maintain brain function and promote brain plasticity. PMID- 12086748 TI - The placebo effect in Parkinson's disease. AB - The biochemical bases of the placebo effect are still incompletely known. We show here that the placebo effect in Parkinson's disease is due, at least in part, to the release of dopamine in the striatum. We propose that the placebo effect might be related to reward mechanisms. The expectation of reward (i.e. clinical benefit) seems to be particularly relevant. According to this theory, brain dopamine release could be a common biochemical substrate for the placebo effect encountered in other medical conditions, such as pain and depression. Other neurotransmitters or neuropeptides, however, are also likely to be involved in mediating the placebo effect (e.g. opioids in pain disorders, serotonin in depression). PMID- 12086749 TI - Getting excited about GABA and sex differences in the brain. AB - In a unique reversal of function, the ubiquitous and dominant inhibitory neurotransmitter, GABA, serves as the major source of neuronal excitation in the developing brain. Opening of voltage-gated Ca(2+) channels following GABA(A) receptor-mediated depolarization translates the action of GABA into trophic responses, such as changes in cell death and synaptic patterning. During a perinatal sensitive period, steroid hormones, in particular estradiol, modulate these cellular responses to differentiate male and female brains. Profound sex differences are found in the volumes of specific nuclei, and the frequencies and types of synapses, in estradiol-concentrating brain regions. Recent evidence suggests that the mechanism of estradiol action involves enhancing, and extending the duration of, the developmental excitatory effects of GABA, resulting in divergence of the signal transduction pathways activated in males versus females. This is the first identification of a pivotal point in brain development that dictates the sex differences evident in adult physiology and behavior. PMID- 12086750 TI - Cytotoxic T lymphocytes in autoimmune and degenerative CNS diseases. AB - Cytotoxic T lymphocytes (CTLs) with a CD8(+) phenotype have the potential to recognize and attack major histocompatibility complex (MHC) class I-expressing brain cells. Most brain cells, including neurons, can be stimulated to present peptides to CD8(+) CTLs by MHC class I molecules, and are susceptible to CTL mediated cytotoxicity in culture. In disease-affected brain parenchyma, CD8(+) CTLs outnumber other T-cell subtypes. They show clonal expansion in several inflammatory and degenerative CNS diseases, such as multiple sclerosis (MS), virus-induced inflammatory brain diseases and paraneoplastic neurological disorders. In MS, damage of axons is closely linked to the CD8(+) CTLs, and protection against CTL-mediated damage should be considered as a new therapeutic approach in MS and other neuroinflammatory diseases. PMID- 12086751 TI - Chronic pain and medullary descending facilitation. AB - Chronic pain, whether the result of nerve trauma or persistent inflammation, is a debilitating condition that exerts a high social cost in terms of productivity, economic impact and quality of life. Currently available therapies yield limited success in treating such pain, suggesting the need for new insight into underlying mechanism(s). Here, we examine the likelihood that sustained activation of descending modulatory pathways that facilitate pain transmission could underlie some states of chronic pain. Such activation of descending facilitatory pathways might be the result of neuroplastic changes that occur at medullary sites in response to persistent input of pain signals. Understanding the mechanisms of descending facilitation and the spinal effects of such discharge could provide new insights into the modulation of chronic pain. PMID- 12086753 TI - Foie gras, fine words, and failure--just another UN summit. PMID- 12086754 TI - Therapeutic angiogenesis: a step forward in intermittent claudication. PMID- 12086755 TI - Insulin-like growth factor-II imprinting in cancer. PMID- 12086756 TI - MMR vaccination, ileal lymphoid nodular hyperplasia, and pervasive develop-mental disorder. PMID- 12086757 TI - Therapeutic angiogenesis with recombinant fibroblast growth factor-2 for intermittent claudication (the TRAFFIC study): a randomised trial. AB - BACKGROUND: Recombinant fibroblast growth factor-2 (rFGF-2) improves perfusion in models of myocardial and hindlimb ischaemia. We investigated whether one or two doses of intra-arterial rFGF-2 improves exercise capacity in patients with moderate-to-severe intermittent claudication. METHODS: 190 patients with intermittent claudication caused by infra-inguinal atherosclerosis were randomly assigned (1:1:1) bilateral intra-arterial infusions of placebo on days 1 and 30 (n=63); rFGF-2 (30 microg/kg) on day 1 and placebo on day 30 (single-dose, n=66); or rFGF-2 (30 microg/kg) on days 1 and 30 (double-dose, n=61). Primary outcome was 90-day change in peak walking time. Secondary outcomes included ankle brachial pressure index and safety. The main analysis was per protocol. FINDINGS: Before 90 days, six patients had undergone peripheral revascularisation and were excluded, and ten withdrew or had missing data. 174 were therefore assessed for primary outcome. Peak walking time at 90 days was increased by 0.60 min with placebo, by 1.77 min with single-dose, and by 1.54 min with double-dose. By ANOVA, the difference between groups was p=0.075. In a secondary intention-to treat analysis, in which all 190 patients were included, the difference was p=0.034. Pairwise comparison showed a significant difference between placebo and single-dose (p=0.026) but placebo and double-dose did not differ by much (p=0.45). Serious adverse events were similar in all groups. INTERPRETATION: Intra-arterial rFGF-2 resulted in a significant increase in peak walking time at 90 days; repeat infusion at 30 days was no better than one infusion. The findings of TRAFFIC provide evidence of clinical therapeutic angiogenesis by intra arterial infusion of an angiogenic protein. PMID- 12086758 TI - Effect of highly active antiretroviral therapy on incidence of tuberculosis in South Africa: a cohort study. AB - BACKGROUND: Studies of the effect of highly active antiretroviral therapy (HAART) on the risk of HIV-1-associated tuberculosis have had variable results. We set out to determine the effect of HAART on the risk of tuberculosis in South Africa. METHODS: We compared the risk of tuberculosis in 264 patients who received HAART in phase III clinical trials and a prospective cohort of 770 non-HAART patients who were attending Somerset Hospital adult HIV clinic, University of Cape Town, between 1992 and 2001. Poisson regression models were fitted to determine risk of tuberculosis; patients were stratified by CD4 count, WHO clinical stage, and socioeconomic status. FINDINGS: HAART was associated with a lower incidence of tuberculosis (2.4 vs 9.7 cases per 100 patient-years, adjusted rate ratio 0.19 [95% CI 0.9-0 38]; p<0.0001). This finding was apparent across all strata of socioeconomic status, baseline WHO stage, and CD4 count, except in patients with CD4 counts of more than 350 cells/microL. The number of tuberculosis cases averted by HAART was greatest in patients with WHO stage 3 or 4 (18.8 averted cases per 100 patient-years, adjusted rate ratio 0. 22 [0.09-0.41]; p=0.03) and in those with CD4 counts of less than 200 cells/microL (14.2 averted cases per 100 patient-years, adjusted rate ratio 0.18 [0.07-0.47]; p,0.0001). INTERPRETATION: HAART reduced the incidence of HIV-1-associated tuberculosis by more than 80% (95% CI 62-91) in an area endemic with tuberculosis and HIV-1. The protective effect of HAART was greatest in symptomatic patients and those with advanced immune suppression. PMID- 12086759 TI - Aggressive conventional chemotherapy compared with high-dose chemotherapy with autologous haemopoietic stem-cell transplantation for relapsed chemosensitive Hodgkin's disease: a randomised trial. AB - BACKGROUND: High-dose chemotherapy followed by transplantation of autologous haemopoietic stem cells (BEAM-HSCT) is frequently used to treat patients with relapsed Hodgkin's disease. We aimed to compare this treatment with conventional aggressive chemotherapy without stem-cell transplantation (Dexa-BEAM). METHODS: 161 patients between 16 and 60 years of age with relapsed Hodgkin's disease were randomly assigned two cycles of Dexa-BEAM (dexamethasone and carmustine, etoposide, cytarabine, and melphalan) and either two further courses of Dexa-BEAM or high-dose BEAM and transplantation of haemopoietic stem cells. Only patients with chemosensitive disease (complete or partial remission after two courses of Dexa-BEAM) proceeded to further treatment. The primary endpoint was freedom from treatment failure for patients with chemosensitive disease. Analysis was per protocol. FINDINGS: 17 patients were excluded from the study after randomisation (ten given Dexa-BEAM and seven given BEAM-HSCT). Median follow-up was 39 months (IQR 3-78). Freedom from treatment failure at 3 years was significantly better for patients given BEAM-HSCT (55%) than for those on Dexa-BEAM (34%; difference 21%, 95% CI -39.87 to -2.13; p=0.019). Overall survival of patients given either treatment did not differ significantly. INTERPRETATION: High-dose BEAM and transplantation of haemopoietic stem cells improves freedom from treatment failure in patients with chemosensitive first relapse of Hodgkin's disease irrespective of length of initial remission. PMID- 12086760 TI - Acarbose for prevention of type 2 diabetes mellitus: the STOP-NIDDM randomised trial. AB - BACKGROUND: The worldwide increase in type 2 diabetes mellitus is becoming a major health concern. We aimed to assess the effect of acarbose in preventing or delaying conversion of impaired glucose tolerance to type 2 diabetes. METHODS: In a multicentre, placebo-controlled randomised trial, we randomly allocated patients with impaired glucose tolerance to 100 mg acarbose or placebo three times daily. The primary endpoint was development of diabetes on the basis of a yearly oral glucose tolerance test (OGTT). Analyses were by intention to treat. FINDINGS: We randomly allocated 714 patients with impaired glucose tolerance to acarbose and 715 to placebo. We excluded 61 (4%) patients because they did not have impaired glucose tolerance or had no postrandomisation data. 211 (31%) of 682 patients in the acarbose group and 130 (19%) of 686 on placebo discontinued treatment early. 221 (32%) patients randomised to acarbose and 285 (42%) randomised to placebo developed diabetes (relative hazard 0.75 [95% CI 0.63 0.90]; p=0.0015). Furthermore, acarbose significantly increased reversion of impaired glucose tolerance to normal glucose tolerance (p<0.0001). At the end of the study, treatment with placebo for 3 months was associated with an increase in conversion of impaired glucose tolerance to diabetes. The most frequent side effects to acarbose treatment were flatulence and diarrhoea. INTERPRETATION: Acarbose could be used, either as an alternative or in addition to changes in lifestyle, to delay development of type 2 diabetes in patients with impaired glucose tolerance. PMID- 12086761 TI - Rare site for an intramuscular lipoma. PMID- 12086763 TI - Systemic mastocytosis and recurrent anaphylactic shock. PMID- 12086762 TI - Endothelial injury mediated by cytotoxic T lymphocytes and loss of microvessels in chronic graft versus host disease. AB - BACKGROUND: Vascular endothelial cells form the interface between recipient tissues and circulating alloreactive donor T cells after allogeneic stem cell transplantation. Vascular injury has been seen in patients with acute graft versus host disease (GVHD) in the skin. We aimed to see whether vascular injury mediated by cytotoxic T lymphocytes and microvessel loss arises in patients with chronic GVHD. METHODS: We investigated eight patients with acute GVHD and ten with chronic GVHD for signs of endothelial injury and microvessel loss by measurement of von Willebrand factor (vWF) in plasma and blood vessel density in biopsy samples taken from lesional skin. Controls consisted of nine patients without GVHD who survived for longer than 100 days and nine healthy people. Inflammation and endothelial injury were assessed in selected samples by immunostaining for CD8 T cells, activated cytotoxic T lymphocytes, and vascular endothelial cells. FINDINGS: We identified more extensive loss of microvessels in the skin of patients with GVHD (median 66 capillaries/mm(2); IQR 16-98) than of healthy controls (205 capillaries/mm(2); 157-226; p=0.005). Patients with GVHD had higher concentrations of vWF (238%; 168-288) than did those without GVHD (102%; 88-118; p=0.0005). Perivascular CD8 T cell infiltrates in skin correlated with vWF plasma concentrations in patients with GVHD (p=0.01), and activated cytotoxic T lymphocytes and endothelial injury were present in these same samples. INTERPRETATION: Host endothelial cells are a target of alloreactive donor cytotoxic T lymphocytes. Substantial blood vessel loss may lead to impaired blood perfusion and tissue fibrosis, the hallmark lesion of chronic GVHD. PMID- 12086764 TI - Small fibre neuropathy in sarcoidosis. AB - Some patients with sarcoidosis have unexplained pain and dysaesthesia. We did quantitative sensory testing in 31 sarcoidosis patients with pain or autonomic dysfunction. 25 patients had reduced warmth sensitivity, cold sensitivity, or both. Intraepidermal nerve fibre density (IENFD) was measured in punch biopsy skin samples in seven consecutive patients. All seven patients had reduced IENFD compared with controls, which confirmed the presence of small fibre neuropathy in these patients. Some patients with sarcoidosis may have small fibre neuropathy with autonomic involvement. PMID- 12086765 TI - Association of antipsychotic drug-induced weight gain with a 5-HT2C receptor gene polymorphism. AB - A side-effect of treatment with antipsychotic drugs for schizophrenia is increased body fat, which leads to further morbidity and poor adherence to treatment. The 5-hydroxytryptamine 2C receptor (5-HT2C) has been associated with this effect; we aimed to establish whether a genetic polymorphism of the promoter region of this receptor affects weight gain after drug treatment in first-episode patients with schizophrenia. We noted significantly less weight gain in patients with the -759T variant allele (p=0.0003) than in those without this allele, who were more likely to have substantial (>7%) weight gain (p=0.002). We have identified a genetic factor that is associated with antipsychotic drug-induced weight gain. PMID- 12086767 TI - The causal links between stress and burnout in a longitudinal study of UK doctors. AB - Burnout and stress are common, linked problems in health-care workers. We aimed to clarify their causal associations. We assessed stress and the three components of burnout (emotional exhaustion, depersonalisation, and low personal accomplishment) using structural equation modelling in a 3-year longitudinal study of a representative sample of 331 UK doctors. Emotional exhaustion and stress showed reciprocal causation: high levels of emotional exhaustion caused stress (beta=0 189), and high levels of stress caused emotional exhaustion (beta=0 175). High levels of personal accomplishment increased stress levels (beta=0 080), whereas depersonalisation lowered stress levels (beta=-0 105). PMID- 12086766 TI - Aminopeptidase P in individuals with a history of angio-oedema on ACE inhibitors. AB - Angio-oedema is a rare but potentially life threatening side-effect of angiotensin-converting-enzyme (ACE) inhibitor treatment. Identification of individuals at risk of this adverse effect is not possible. Angio-oedema is associated with raised concentrations of bradykinin, which is mainly inactivated by ACE. We assessed the plasma activity of two other enzymes that catabolise bradykinin (aminopeptidase P and carboxypeptidase N) in 39 hypertensive patients with a history of angio-oedema during ACE inhibitor treatment and in 39 hypertensive patients who had never had ACE inhibitor associated side-effects. Patients with previous angio-oedema had a lower plasma activity of aminopeptidase P than did those who never presented with angio-oedema (p=0 003). Our data suggest that low plasma concentrations of aminopeptidase P could be a predisposing factor for development of angio-oedema in patients treated with ACE inhibitors. PMID- 12086768 TI - Sponsor of US cloning bill signals shift in tactics. PMID- 12086769 TI - US states file lawsuit against Bristol-Myers Squibb. PMID- 12086771 TI - Collated evidence confirms the value of atrial fibrillation prevention. PMID- 12086772 TI - US-led health programme breaks down boundaries in Middle East. PMID- 12086773 TI - Indian children with epilepsy do not have access to specific services, says report. PMID- 12086774 TI - FDA allows controversial bowel drug back on to market. PMID- 12086776 TI - HIV cases in S Africa decreasing. PMID- 12086777 TI - Financial plans spread through US health care. PMID- 12086778 TI - HIV-1/AIDS and maternal and child health in Africa. AB - Every day, 1900 children acquire HIV-1 infection from their mother in Africa. The 25-45% risk of mother-to-child transmission can be reduced in several ways: prevention of sexual transmission for women of child-bearing age, access to HIV-1 testing, reduction of unwanted pregnancies by education of HIV-1-infected women, and antiretroviral-based prevention. All antiretroviral regimens of proven efficacy can be used in a minimum package of care for HIV-1-infected pregnant women. At present, programmes in 13 countries reach less than 3% of HIV-1 infected African women. 35-59% of African children infected with HIV-1 die by their second birthday. Infectious complications are preventable by primary prophylaxis with co-trimoxazole. A rapid scaling-up and comprehensive continuum of care is needed for all members of affected families, including access to antiretroviral treatment and community-based responses to the increasing number of orphans. Prevention of mother-to-child transmission should become a universal standard of care in Africa, and research should continue to reduce the transmission risk to well below 5%. PMID- 12086779 TI - Death of a child. PMID- 12086780 TI - Differential pricing of drugs: a role for cost-effectiveness analysis? AB - Internationally, the high costs of pharmaceutical products limit access to treatment. The principle of differential pricing is that drug prices should vary according to some measure of affordability. How differential prices should be determined is, however, unclear. Here we describe a method whereby differential prices for essential drugs could be derived in countries of variable national wealth, and, using angiotensin-converting enzyme inhibitors provide an example of how the process might work. Indicative prices for drugs can be derived by cost effectiveness analysis that incorporates a measure of national wealth. Such prices could be used internationally as a basis of differential price negotiations. PMID- 12086781 TI - Tobacco money: up in smoke? PMID- 12086782 TI - Autism, bowel inflammation, and measles. PMID- 12086783 TI - Autism, bowel inflammation, and measles. PMID- 12086784 TI - Autism, bowel inflammation, and measles. PMID- 12086785 TI - Autism, bowel inflammation, and measles. PMID- 12086786 TI - Autism, bowel inflammation, and measles. PMID- 12086787 TI - Multidrug resistance and response to antiretroviral treatment. PMID- 12086789 TI - Intestinal involvement in children with Behcet's disease. PMID- 12086790 TI - Dynamic epidemiology of group A streptococcal serotypes. PMID- 12086791 TI - Near-death experiences. PMID- 12086792 TI - Near-death experiences. PMID- 12086793 TI - Near-death experiences. PMID- 12086794 TI - Near-death experiences. PMID- 12086795 TI - Blood pressure and cardiovascular risk in the HOPE study. PMID- 12086798 TI - Climate change: the new bioterrorism. PMID- 12086796 TI - Blood pressure and cardiovascular risk in the HOPE study. PMID- 12086799 TI - Is the NICE process flawed? PMID- 12086800 TI - Is the NICE process flawed? PMID- 12086801 TI - Striking reduction in medical junk mail. PMID- 12086803 TI - Hospital life pictured en route. PMID- 12086804 TI - Bone bobbins and metal buttons. PMID- 12086809 TI - The sensory function of ligaments. PMID- 12086810 TI - Spinal and supraspinal effects of activity in ligament afferents. AB - In this paper available knowledge on effects from joint and ligament afferents on spinal neurones and pathways are briefly reviewed, and possible functional implications discussed. Ligament afferents may contribute to joint stability, muscle coordination and proprioception through direct polysynaptic reflex effects onto ascending pathways and skeletomotoneurones, and/or indirectly via reflex actions on the gamma-muscle spindle system. Theoretical and experimental evidence indicate that ligament afferents, together with afferents from other joint structures, muscles and the skin, provide the CNS with information on movements and posture through ensemble coding mechanisms, rather than via modality specific private pathways. The existence and functional relevance of ligamentomuscular protective reflexes, that are triggered when the ligament is threatened by potentially harmful loads, has been seriously questioned. It seems more likely that peripheral sensory inputs from ligament afferents participate in a continuous control of the muscle activity through feedforward, or preprogramming, mechanisms. In line with these ideas it has been suggested that ligament mechanoreceptors have an important role in muscle coordination and in the reflex regulation of the functional joint stability, by contributing to the preprogramming of the muscle stiffness through reflex modulation of the gamma muscle spindle system. PMID- 12086811 TI - Cruciate ligament reflexes. AB - The idea of muscular reflexes elicited from sensory nerves of the cruciate ligaments is more than 100 years old, but the existence of such reflexes has not been proven until the recent two decades. First in animal experiments, a muscular excitation could be elicited in the hamstrings when the anterior cruciate ligament (ACL) was pulled, and tension in the ligament caused activity of the gamma motor neurones of the muscles around the knee. Impulses from the sensory nerves in ACL were activated during motion of the knee, in particular overstretching and combined extension and rotation. In humans, proprioception in the knee is decreased after ACL rupture. By mechanical or electrical stimulation of the ACL, an excitation in the hamstrings muscles can be elicited. During muscular activity, stimulation of the ACL or PCL results in a clear inhibition of the ongoing activity, both during static isometric and isokinetic muscle work, and also during dynamic activity (gait). This inhibitory reflex subjectively resembled giving way. The latency of the reflex was short in animals (about 3 ms) and long in humans (60-120 ms), probably caused by differences in the experimental setup and between species. The long latency in humans makes it unlikely that it is a directly protective reflex. Instead it may be involved in the updating of motor programs. Further research may characterize the reflex in details and map its pathways. The existence of this reflex indicate that the cruciate ligaments have an afferent function, which influences knee dynamics. PMID- 12086812 TI - Shoulder reflexes. AB - Dynamic shoulder stability is dependent on muscular coordination and sensory inputs. In the shoulder, mechanoreceptors are found in the coracoacromial ligament, the rotator cuff tendons, the musculotendinous junctions of the rotator cuff and in the capsule. The number of receptors in the capsule is small compared to the number in the other shoulder structures. Proprioceptive information from numerous receptors in muscles and tendons is mediated via fast conducting nervefibers and probably contribute more to kinaestethic sensation than information from capsule and ligaments. Therefore it seems likely that the joint receptors have a more distinct role for the kinaestethic sense than muscle receptors. In cats a direct reflex from the afferents innervating the shoulder to the muscles around the shoulder has been presented. The reflex had an extremely short latency (2.7-3.1 ms). In man, a very long latency (300 ms) excitatory reflex has been found when nerves in the capsule were stimulated electrically during shoulder surgery. In addition, when the anterior-inferior capsule was excited in conscious humans with modest amplitude electrical stimuli during muscle activity, a strong inhibition was found with an average latency of 33 ms. Stimulation of the sensory nerves in the coracoacromial ligament has also been found to modify muscle activity strongly. Even though our understanding of the control of shoulder motion is incomplete, it is clear that sensory inputs can strongly modify muscle activity around the shoulder. This has implications for rehabilitation and shoulder surgery. PMID- 12086813 TI - Reflex from the ankle ligaments of the feline. AB - It was found that two to three articular branches of the tibial nerve innervate the medial ligament of the feline ankle. No innervation was found to the laterial ligament. Supramaximal electrical stimulation of the articular nerves was found to elicit electromyographic (EMG) activity in the intrinsic muscles of the foot. EMG activity was not found in any of the calf muscles which cross the ankle. The average time delay from stimulus to EMG activity was 3.8 ms, indicating that a fast, bisynaptic reflex is active, probably for purposes of preventing or correcting foot eversion to maintain joint stability. PMID- 12086814 TI - Sensori-motor control of the uninjured and injured human ankle. AB - This review focuses on the role of sensori-motor function in the healthy as well as the functionally unstable ankle. The concept functional ankle instability--a widely used term, which has no universally agreed upon definition-as well as the sources of peripheral afferent information measured with different sensori-motor tests are discussed. The protective mechanisms against sudden ankle inversion are reviewed, and models that directly connect deficits in kinaesthesia and peroneal reflex reaction to an increased risk of sustaining unprovoked ankle inversion injuries are presented. PMID- 12086815 TI - Consequences of a ligament injury on neuromuscular function and relevance to rehabilitation - using the anterior cruciate ligament-injured knee as model. AB - The purpose of this article is to survey current knowledge concerning the consequences of a ligament injury on neuromuscular function and its relevance to rehabilitation, in relation to clinical practice. Although it deals with the ACL injured knee, these views may also apply to other joints. The effects of a ligament injury on neuromuscular function are first considered - i.e., proprioception, postural control, muscle strength, functional performance, movement and activation pattern, central mechanisms, motor control and learning. The treatment and effects of rehabilitation on neuromuscular function are then discussed. The survey is concluded by discussing the clinical significance. PMID- 12086816 TI - Afferent feedback in the control of human gait. AB - Sensory activity contributes to motor control in two fundamentally different ways. It may mediate 'error signals' following sudden external perturbations and it may contribute to the pre-programmed motoneuronal drive. Here we review data, which illustrate these two functions of sensory feedback in relation to human walking. When ankle plantarflexors are unloaded in the stance phase there is a sudden decrease in the sensory activity in muscle and tendon afferents from the active muscles. This decrease in sensory activity results in a drop in EMG activity recorded from the soleus muscle, which demonstrates that the sensory activity contributes importantly to the activation of the muscles. Data suggests that a spinal pathway from gr. II muscle afferents is responsible for this positive feedback contribution to the motoneuronal drive during walking. When cutaneous nerves from the foot are stimulated in the early swing phase of walking a late reflex response may be observed in the tibialis anterior muscle. This reflex may help to ensure that the foot is lifted effectively over an obstacle. Data suggest that this reflex response is at least partly mediated by a transcortical reflex pathway. It seems to be important that reactions to external perturbations are integrated at a supraspinal level during human walking. PMID- 12086817 TI - Sensorimotor control of the spine. AB - The spinal viscoelastic structures including disk, capsule and ligaments were reviewed with special focus on their sensory motor functions. Afferent capable of monitoring proprioceptive and kinesthetic information are abundant in the disc, capsule and ligament. Electrical stimulation of the lumbar afferents in the discs, capsules and ligaments seem to elicit reflex contraction of the multifidus and also longissimus muscles. The muscular excitation is pronounced in the level of excitation and with weaker radiation 1 to 2 levels above and below. Similarly, mechanical stimulation of the spinal viscoelastic tissues excites the muscles with higher excitation intensity when more than one tissue (ligaments and discs for example) is stimulated. Overall, it seems that spinal structures are well suited to monitor sensory information as well as to control spinal muscles and probably also provide kinesthetic perception to the sensory cortex. PMID- 12086818 TI - Modification of the swern oxidation: use of a recyclable, polystyrene bound sulfoxide. AB - A method has been developed for the oxidation of sulfides to sulfoxides on polystyrene resin. The polystyrene bound sulfoxide may be used in Swern oxidation reactions, and the used reagent may be regenerated by oxidation with tert butylhydrogen peroxide. PMID- 12086819 TI - Polymer-supported ferrocenyl oxazolines for the catalyzed highly enantioselective phenyl transfer to aldehydes. AB - A new chiral MeO-PEG-supported ferrocenyl oxazoline was synthesized and successfully employed in the enantioselective phenyl transfer to aldehydes. The products were obtained in high yields and with excellent enantioselectivities (up to 97% ee). Furthermore, the recovery of the ferrocene was facile, and catalyst efficiency was maintained in subsequent reactions. PMID- 12086820 TI - Synthesis of a novel, recyclable, solid-phase acylating reagent. AB - In this paper, we describe the synthesis of N-(6-cyano-1,3-dimethyl-2,4-dioxo-5 substituted-1,3-dihydropyridino[2,3-d] pyrimidin-7-yl)imides 1. We will show the synthesis of 1 using both conventional heating and microwave techniques. In addition, the imide was attached to polystyrene and this immobilized imide was equally as effective at acylating various primary and secondary amines as its solution-phase counterpart. PMID- 12086822 TI - Solid-phase preparation of amides using N-acylbenzotriazoles. AB - Wang resin linked amines were efficiently converted into amides using acylbenzotriazoles. Cleavage of resins gave the desired amides 7Aa-Gf in 30-99% yields with good to excellent purities. PMID- 12086821 TI - Towards more chemically robust polymer-supported chiral catalysts for the reactions of aldehydes with dialkylzincs. AB - N-Methyl-alpha,alpha-diphenyl-L-prolinol derivatives with para-bromo substituents in one or both of the phenyl rings are easily bound to crosslinked polystyrene beads containing phenylboronic acid residues using Suzuki reactions. When the products were used as catalysts for the reactions of aldehydes with diethylzinc in toluene at 20 degrees C, the alcohols were produced in chemical yields >90% and with ees of upto 94%. The better of the two supported catalysts gave ees only 0-9% lower than those obtained with the corresponding soluble catalyst. One of the supported catalysts was recycled successfully nine times. PMID- 12086823 TI - A polymer-supported [1,3,2]oxazaphospholidine for the conversion of isothiocyanates to isocyanides and their subsequent use in an ugi reaction. AB - The design and synthesis of a new polymer supported reagent for the clean conversion of isothiocyanates to isocyanides under microwave conditions was accomplished. The structurally diverse isocyanides generated were used in an Ugi 3CC, allowing the rapid generation of 2-isoindolinone-7-carboxamide analogues. PMID- 12086824 TI - 2,7-Di-tert-butyl-Fmoc-P-OSu: a new polymer-supported reagent for the protection of the amino group. AB - The polymer-supported (2,7-di-tert-butyl-9-fluorenyl)methyl succinimidyl carbonate (Dtb-Fmoc-P-OSu), derived from (2,7-di-tert-butyl-9-fluorenyl)methyl chloroformate (Fmoc-Cl) and a polymeric N-hydroxysuccinimide (P-HOSu), has been used for the preparation of Dtb-Fmoc-protected amines and amino acids. After the N-protection reaction, the liberated P-HOSu can be recovered and reused. This Dtb Fmoc-protection improves the solubility of the Fmoc-protected analogues. PMID- 12086825 TI - Improvement of ligand economy controlled by polymer morphology: the case of polymer-supported bis(oxazoline) catalysts. AB - A functionalized chiral bis(oxazoline) is used as a chiral monomer in polymerization reactions leading to homo- and copolymers of different morphology. Polymers with a high content of chiral monomer lead to enantioselectivities that are higher than those obtained with the soluble ligand, but the chiral ligand is not used in an optimal way. A hyperbranched polymer, obtained by using a hexavinyldendrimer as the cross-linker, leads to the same enantioselectivities with a more efficient use of the chiral ligand. PMID- 12086826 TI - Asymmetric hydroformylation with highly crosslinked polystyrene-supported (R,S) BINAPHOS-Rh(I) complexes: the effect of immobilization position. AB - A new class of polymer-supported (R,S)-BINAPHOS 1e in which the parent BINAPHOS has two alkoxy-substituents at the 3-positions of the phenyls, has been synthesised. Using its Rh(I) complex, asymmetric hydroformylation of olefins proceeded with higher enantioselectivity in some cases compared to the conventional polymer-supported 1c. PMID- 12086827 TI - Solid-phase C-C and C-O bond forming reactions using 'traceless' pi-arene chromium linkers. AB - The scope of solid-phase reactions utilizing a 'traceless' pi-arene-chromium linker has been expanded to include the addition of organo-lithium reagents to a supported ester group, generation and alkylation of an enolate anion, and substitution via the Mitsunobu reaction. Immobilization of ortho-disubstituted polyfunctionalized arene chromium dicarbonyl species onto the solid support and subsequent, selective transformation is also reported. PMID- 12086828 TI - Polymer-assisted Horner-Emmons olefination using PASSflow reactors: pure products without purification. AB - A PASSflow protocol for the Horner-Emmons olefination of aldehydes using polymer bound hydroxide ions in flowthrough reactors is presented which allows preparation of alkenes in very high yield with minimal purification. PMID- 12086829 TI - Heterogeneous C-C coupling and polymerization catalysts prepared by ROMP. AB - This contribution summarizes the latest developments in the area of catalytic supports prepared via ring-opening metathesis polymerization (ROMP). In particular, the synthesis of heterogeneous catalytic systems active in Pd mediated C-C coupling reactions such as Heck, Sonogashira-Hagihara and Suzuki couplings, as well as in ruthenium-mediated olefin metathesis reactions will be summarized. The general concept for the synthesis of these supports will be outlined in detailed. PMID- 12086830 TI - Polymer-supported catalysts: enantioselective hydrogenation and hydrogen transfer reduction. AB - Different types of heterogenized catalysts were involved in asymmetric reactions. Hydrogen transfer reduction was performed with amino alcohols derived from poly((S)-(GMA-co-EGDMA or DVB)) and hydrogenation with BINAP grafted onto PEG or copolymerized with isocyanates as ligands. Attempts of catalysts recycling are reported. PMID- 12086831 TI - Polymer-bound 1-aryl-3-alkyltriazenes as modular ligands for catalysis. Part 1: synthesis and metal coordination. AB - A range of di- and tri-substituted triazenes have been synthesized from a polymer supported diazonium salt and various primary and secondary amines. The triazenes obtained were treated with transition metal salts to form polymer-supported metal complexes in firstly a general screen and then in a specific manner. PMID- 12086832 TI - Polymer-bound 1-aryl-3-alkyltriazenes as modular ligands for catalysis. Part 2: screening immobilized metal complexes for catalytic activity. AB - A range of polymer-supported triazenes and their metal-bound analogues were screened for use in catalysis. Fe, Cu and Zr complexes were, together with the polymer-supported triazene ligand alone, screened in the addition of Et(2)Zn to benzaldehyde. A supported Pd triazene complex was screened for activity in Suzuki and Sonogashira reactions and a supported Ru triazene complex screened for transfer hydrogenation. PMID- 12086833 TI - Preparation of polymer-supported benzyllithium reagents. AB - Cross-linked polystyrenes (2, 8, and 11) possessing pendant double bonds conjugated with benzene ring were prepared by both polymerization method and chemical modification of pre-formed polymers. These double bonds were readily lithiated with n-butyllithium to give the benzyllithium species on the polymer. The polymer supported benzylic anions (3 and 12) are a versatile polymeric organometallic reagent that reacted with various kinds of electrophiles. PMID- 12086834 TI - Immobilized oxazoline-containing ligands in asymmetric catalysis--a review. AB - Metal complexes of chiral oxazoline derivatives immobilized on soluble as well as insoluble supports serve as versatile asymmetric catalysts in a variety of applications. In a few cases recovery and reuse of the chiral ligands have been achieved. PMID- 12086835 TI - Polymer-supported reagents for methylphosphorylation and phosphorylation of carbohydrates. AB - Two polymer-supported reagents for methylphosphorylation and phosphorylation of carbohydrates have been developed. p-Hydroxybenzyl alcohol and beta mercaptoethanol were immobilized on cross-linked divinylbenzene-polystyrene copolymer and conjugated with methyl N,N-diisopropylchlorophosphoramidite. Carbohydrates were reacted with polymer-bound phosphitylating reagents. Further oxidation, with or without the methoxy group deprotection, and cleavage yielded methylphosphorylated or phosphorylated carbohydrates, respectively. PMID- 12086836 TI - Polymer-supported chiral catalysts with positive support effects. AB - In this paper, we discuss the rational design of polymeric catalysts and the positive effect of polymer supports on the catalytic asymmetric reactions. The attachment of chiral catalysts to soluble polymers, particularly dendritic polymers, offered a potential combination of the advantages of homogeneous and heterogeneous asymmetric catalysis. PMID- 12086837 TI - A solid-supported phosphine-free ruthenium alkylidene for olefin metathesis in methanol and water. AB - The synthesis and olefin metathesis activity in protic solvents of 7, a phosphine free ruthenium alkylidene bound to a hydrophilic solid support are reported. This heterogeneous catalyst promotes relatively efficient ring closing- and cross metathesis reactions in both methanol and water. The potential utility of homogeneous catalyst 2 for olefin metathesis in methanol is also demonstrated. PMID- 12086838 TI - Development of a new ultraporous polymer as support in organic synthesis. AB - This paper describes the preparation and post-functionalisation of a new polymeric support based on emulsion-derived foams and called polyHIPEs. The remaining pendant vinylic bonds are easily functionalised by a free radical mechanism. The large pores and channels of this material allow an easy access of the reagent in solution toward the grafted species. PolyHIPE-supported thiol, in the presence of an excess of triethylsilane, showed a good activity and selectivity toward reductive cyclisation of 6-bromohex-1-ene and 1-allyloxy-2 bromobenzene. PMID- 12086839 TI - A concise route to triazolobenzodiazepine derivatives via a one-pot alkyne-azide cycloaddition reaction. AB - A new and efficient one-pot synthesis of [1,2,3]triazolo[1,5-a][1,4] benzodiazepin-6(4H)-ones is described starting from readily available anthranilic acids. A small array of the title compounds were assembled via a four-step sequence involving diazotisation, azide addition followed by amide bond formation employing polymer supported carbodiimide and subsequent 1,3-dipolar cycloaddition reaction. PMID- 12086841 TI - Altered methylation patterns in cancer cell genomes: cause or consequence? AB - CpG islands are associated with at least half of all cellular genes and are normally methylation-free. Dense methylation of cytosine residues within islands causes strong and heritable transcriptional silencing. Such silencing normally occurs almost solely at genes subject to genomic imprinting or to X chromosome inactivation. Aberrant methylation of CpG islands associated with tumor suppressor genes has been proposed to contribute to carcinogenesis. However, questions of mechanisms underlying the cancer changes and the precise consequences for tumorigenesis exist in the field, and must continue to be addressed before the importance of abnormalities in genomic methylation patterns in carcinogenesis can be fully understood. In this article, two workers in DNA methylation, one concentrating on cancer biology and the other on developmental biology, address recurrent questions about cancer epigenetics from different perspectives. The goal is to highlight important controversies in the field which can be productive targets of ongoing and future research. PMID- 12086842 TI - (Re-)Engineering tumor cell-selective replicating adenoviruses: a step in the right direction toward systemic therapy for metastatic disease. AB - An approach combining redundant controls to restrict the productive infection of adenoviruses to cells that are disrupted in the pRb pathway-a hallmark of human cancer-has resulted in a novel oncolytic virus that may be well suited for systemic administration to treat metastatic disease. PMID- 12086843 TI - Senescence: a companion in chemotherapy? AB - Mouse lymphoma model points to an unsuspected role of drug-induced "senescence." PMID- 12086844 TI - p53 leans on its siblings. AB - Despite the common assumption that p53 by itself can induce apoptosis, results of a recent study implicate the homologous genes p63 and p73 in p53-mediated programmed cell death. PMID- 12086845 TI - Cancer Molecular Analysis Project: weaving a rich cancer research tapestry. AB - The Cancer Molecular Analysis Project (CMAP) of the NCI is integrating diverse cancer research data to elucidate fundamental etiologic processes, enable development of novel therapeutic approaches, and facilitate the bridging of basic and clinical science. PMID- 12086846 TI - Focus on breast cancer. PMID- 12086848 TI - Selectively replicating adenoviruses targeting deregulated E2F activity are potent, systemic antitumor agents. AB - We have engineered a human adenovirus, ONYX-411, that selectively replicates in human tumor cells, but not normal cells, depending upon the status of their retinoblastoma tumor suppressor protein (pRB) pathway. Early and late viral gene expression as well as DNA replication were significantly reduced in a functional pRB-pathway-dependent manner, resulting in a restricted replication profile similar to that of nonreplicating adenoviruses in normal cells both in vitro and in vivo. In contrast, the viral life cycle and tumor cell killing activity of ONYX-411 was comparable to that of wild-type adenovirus following infection of human tumor cells in vitro as well as after systemic administration in tumor bearing animals. PMID- 12086849 TI - Elevated levels of IGF-1 receptor convey invasive and metastatic capability in a mouse model of pancreatic islet tumorigenesis. AB - In a prototypical model of multistage tumorigenesis involving pancreatic islets in RIP1-Tag2 transgenic mice, activation of insulin-like growth factor II (IGF II) was previously shown to serve as a survival factor that inhibited apoptosis. Now IGF-1R, the receptor tyrosine kinase for IGF-II, has been found to be variably upregulated, first uniformly in dysplastic and angiogenic progenitors and then focally at the margins and in invasive regions of carcinomas. When the levels of IGF-1R were forcibly elevated throughout islet tumorigenesis, progression was accelerated at all stages in the pathway, although apoptosis was not differentially suppressed. Notably, encapsulated tumors were absent; instead, invasive carcinomas with downregulated E-cadherin were prevalent, and the majority of mice had local lymph node metastasis. PMID- 12086850 TI - Pathway-specific tumor suppression. Reduction of p27 accelerates gastrointestinal tumorigenesis in Apc mutant mice, but not in Smad3 mutant mice. AB - Expression of the cyclin-dependent kinase inhibitor p27(Kip1) (p27) is frequently reduced in human colorectal cancer, and this correlates with poor patient prognosis. To clarify the role of p27 in gastrointestinal (GI) cancer, we measured p27 expression, as well as the effect of germline deletion of p27, in 3 different mouse models of GI neoplasia. p27 expression was frequently reduced in GI tumors arising in 1,2-dimethylhydrazine (DMH) treated mice, and in Apc mutant Min/+ mice, but not in GI tumors arising in Smad3 mutant mice. Germline deletion of p27 resulted in accelerated tumor development and increased tumor cell proliferation in both DMH treated and Min/+ mice, but not in Smad3 mutant mice. p27 deficiency also led to increased adenoma to adenocarcinoma progression. These results indicate that reduction of p27 cooperates with mutations in Apc but not in Smad3 during GI tumorigenesis. Thus, tumor suppression by p27 is contingent on the specific oncogenic pathway that drives tumor development. PMID- 12086852 TI - Positron emission tomography imaging analysis of G2A as a negative modifier of lymphoid leukemogenesis initiated by the BCR-ABL oncogene. AB - G2A is a lymphocyte-expressed G protein-coupled receptor whose genetic ablation results in the development of autoimmunity. Using HSV-TK reporter gene directed positron emission tomography (PET), we demonstrate that prior to any indication of the onset of illness, mice transplanted with BCR-ABL transduced G2A-deficient bone marrow harbor expanded populations of leukemic cells compared to recipients of wild-type bone marrow. The target cell type and anatomical locations of leukemia development are indistinguishable in animals transplanted with G2A+/+ or G2A-/- cells. Shorter disease latency in the G2A-deficient background is associated with an increased rate of cellular expansion. PET can be successfully applied to the temporal and spatial analysis of Bcr-Abl driven leukemic progression and should have utility for the study of other leukemias and lymphomas. PMID- 12086851 TI - DeltaNp63 induces beta-catenin nuclear accumulation and signaling. AB - The P53 homolog p63 encodes multiple proteins with transactivating, apoptosis inducing, and oncogenic activities. We showed that p63 is amplified and that DeltaNp63 isotypes are overexpressed in squamous cell carcinoma (SCC) and enhance oncogenic growth in vitro and in vivo. Moreover, p53 associated with DeltaNp63alpha and mediated its degradation. Here, we report that DeltaNp63 associates with the B56alpha regulatory subunit of protein phosphatase 2A (PP2A) and glycogen synthase kinase 3beta (GSK3beta), leading to a dramatic inhibition of PP2A-mediated GSK3beta reactivation. The inhibitory effect of DeltaNp63 on GSK3beta mediates a decrease in phosphorylation levels of beta-catenin, which induces intranuclear accumulation of beta-catenin and activates beta-catenin dependent transcription. Our results suggest that DeltaNp63 isotypes act as positive regulators of the beta-catenin signaling pathway, providing a basis for their oncogenic properties. PMID- 12086854 TI - Two HIFs may be better than one. AB - HIF-2alpha overexpression directly contributes to renal clear cell tumorigenesis: evidence for HIF as a tumor promoter. PMID- 12086853 TI - The Ewing's sarcoma oncoprotein EWS/FLI induces a p53-dependent growth arrest in primary human fibroblasts. AB - Ewing's sarcoma is associated with a fusion between the EWS and FLI1 genes, forming an EWS/FLI fusion protein. We developed a system for the identification of cooperative mutations in this tumor through expression of EWS/FLI in primary human fibroblasts. Gene expression profiling demonstrated that this system recapitulates many features of Ewing's sarcoma. EWS/FLI-expressing cells underwent growth arrest, suggesting that growth arrest-abrogating collaborative mutations may be required for tumorigenesis. Expression profiling identified transcriptional upregulation of p53, and the growth arrest was rescued by inhibition of p53. These data support a role for p53 as a tumor suppressor in Ewing's sarcoma and demonstrate the use of transcriptional profiling of model systems in the identification of cooperating mutations in human cancer. PMID- 12086855 TI - RUNX: a trilogy of cancer genes. AB - The RUNX family of transcription factors plays pivotal roles during normal development and in neoplasias. Recent data involve RUNX3 as an important tumor suppressor in gastric cancers and pose interesting questions about how perturbed levels and interspecific competition among RUNX family members may contribute to tumorigenesis. PMID- 12086856 TI - The secrets of selective estrogen receptor modulation: cell-specific coregulation. AB - A specific increase in the level of a single coactivator appears to enhance estrogen action with tamoxifen at some gene targets in uterine cells but not breast cells. PMID- 12086858 TI - Zebrafish as a cancer model system. AB - The zebrafish, with its combination of forward genetics and vertebrate biology, has great potential as a cancer model system. PMID- 12086857 TI - Molecular mechanisms of lymphangiogenesis in health and disease. AB - Studies of the last decades have revealed the importance of angiogenesis for normal growth and for the pathogenesis of numerous diseases. Much less studied is lymphangiogenesis, the growth of lymphatic vessels, which drain extravasated fluid, proteins, and cells and transport them back to the venous circulation. Nonetheless, insufficient lymphangiogenesis causes incapacitating lymphedema, while lymphatic growth around tumors may facilitate metastatic spread of malignant cells that ultimately kill the patient. The recent discovery of the key lymphangiogenic factors VEGF-C and VEGF-D and their receptor VEGFR-3 has allowed novel insights into how the lymphatic vessels and blood vessels coordinately grow and affect human disease. In addition, these studies have opened novel diagnostic and therapeutic avenues for the treatment of lymphedema and metastasis. This overview highlights the recent insights and developments in the field of lymphatic vascular research. PMID- 12086859 TI - Focus on colon cancer. PMID- 12086860 TI - Inhibition of HIF is necessary for tumor suppression by the von Hippel-Lindau protein. AB - Inactivation of the von Hippel-Lindau tumor suppressor gene is linked to the development of hereditary (VHL Disease-associated) and sporadic clear cell carcinoma of the kidney. The VHL gene product, pVHL, targets the heterodimeric transcription factor HIF for polyubiquitination, and restoration of pVHL function in VHL(-/-) renal carcinoma cells suppresses their ability to form tumors in nude mice. Here we show that tumor suppression by pVHL can be overridden by a HIF variant that escapes pVHL control. These studies prove that HIF is a critical downstream target of pVHL and establish that activation of HIF target genes can promote tumorigenesis in vivo. PMID- 12086861 TI - The contribution of VHL substrate binding and HIF1-alpha to the phenotype of VHL loss in renal cell carcinoma. AB - Clear-cell renal carcinoma is associated with inactivation of the von Hippel Lindau (VHL) tumor suppressor gene. VHL is the substrate recognition subunit of an E3 ligase, known to target the alpha subunits of the HIF heterodimeric transcription factor for ubiquitin-mediated degradation under normoxic conditions. We demonstrate that competitive inhibition of the VHL substrate recognition site with a peptide derived from the oxygen degradation domain of HIF1alpha recapitulates the tumorigenic phenotype of VHL-deficient tumor cells. These studies prove that VHL substrate recognition is essential to the tumor suppressor function of VHL. We further demonstrate that normoxic stabilization of HIF1alpha alone, while capable of mimicking some aspects of VHL loss, is not sufficient to reproduce tumorigenesis, indicating that it is not the critical oncogenic substrate of VHL. PMID- 12086862 TI - Dissection of angiogenic signaling in zebrafish using a chemical genetic approach. AB - Striking homology between signaling molecules in zebrafish and humans suggests that compounds known to inhibit human kinases may enable a chemical genetic approach to dissect signaling pathways in the zebrafish embryo. We tested this hypothesis using a vascular endothelial growth factor receptor inhibitor, PTK787/ZK222584. Zebrafish embryos treated with this compound lacked all major blood vessels. Overexpression of AKT/PKB, a putative effector of vascular endothelial growth factor signaling, allowed blood vessels to form in the presence of drug. Endothelial cell apoptosis induced by the drug is prevented by increasing AKT/PKB activity, thus establishing the physiological relevance of AKT/PKB in the angiogenic process. This approach allowed us to examine the effects of blood flow and the role of endothelial signals in organogenesis. PMID- 12086863 TI - Epidermal growth factor receptor and Ink4a/Arf: convergent mechanisms governing terminal differentiation and transformation along the neural stem cell to astrocyte axis. AB - Ink4a/Arf inactivation and epidermal growth factor receptor (EGFR) activation are signature lesions in high-grade gliomas. How these mutations mediate the biological features of these tumors is poorly understood. Here, we demonstrate that combined loss of p16(INK4a) and p19(ARF), but not of p53, p16(INK4a), or p19(ARF), enables astrocyte dedifferentiation in response to EGFR activation. Moreover, transduction of Ink4a/Arf(-/-) neural stem cells (NSCs) or astrocytes with constitutively active EGFR induces a common high-grade glioma phenotype. These findings identify NSCs and astrocytes as equally permissive compartments for gliomagenesis and provide evidence that p16(INK4a) and p19(ARF) synergize to maintain terminal astrocyte differentiation. These data support the view that dysregulation of specific genetic pathways, rather than cell-of-origin, dictates the emergence and phenotype of high-grade gliomas. PMID- 12086864 TI - Wnt5a signaling directly affects cell motility and invasion of metastatic melanoma. AB - Gene expression profiling identified human melanoma cells demonstrating increased cell motility and invasiveness. The gene WNT5A best determined in vitro invasive behavior. Melanoma cells were transfected with vectors constitutively overexpressing Wnt5a. Consistent changes included actin reorganization and increased cell adhesion. No increase in beta-catenin expression or nuclear translocation was observed. There was, however, a dramatic increase in activated PKC. In direct correlation with Wnt5a expression and PKC activation, there was an increase in melanoma cell invasion. Blocking this pathway using antibodies to Frizzled-5, the receptor for Wnt5a, inhibited PKC activity and cellular invasion. Furthermore, Wnt5a expression in human melanoma biopsies directly correlated to increasing tumor grade. These observations support a role for Wnt5a in human melanoma progression. PMID- 12086865 TI - Dissecting p53 tumor suppressor functions in vivo. AB - Although the p53 tumor suppressor acts in a plethora of processes that influence cellular proliferation and survival, it remains unclear which p53 functions are essential for tumor suppression and, as a consequence, are selected against during tumor development. Using a mouse model harboring primary, genetically modified myc-driven lymphomas, we show that disruption of apoptosis downstream of p53 by Bcl2 or a dominant-negative caspase 9 confers-like p53 loss-a selective advantage, and completely alleviates pressure to inactivate p53 during lymphomagenesis. Despite their p53-null-like aggressive phenotype, apoptosis defective lymphomas that retain intact p53 genes do not display the checkpoint defects and gross aneuploidy that are characteristic of p53 mutant tumors. Therefore, apoptosis is the only p53 function selected against during lymphoma development, whereas defective cell-cycle checkpoints and aneuploidy are mere byproducts of p53 loss. PMID- 12086866 TI - It's ALL in the diagnosis. AB - The molecular diagnosis of human cancer will hasten the development of treatments tailored to the abnormalities present in each patient's tumor cells. Recent gene expression profiling studies of pediatric acute lymphoblastic leukemia (ALL) suggest that the molecular diagnosis of these diseases is right around the corner. PMID- 12086867 TI - The search for predictive patterns in ovarian cancer: proteomics meets bioinformatics. AB - Proteomic patterns in serum that discriminate between malignant and benign ovaries may provide a powerful tool for screening in high risk women. PMID- 12086868 TI - Looking for a good endothelial address. AB - The first in vivo screening of a peptide library of the human vasculature opens new possibilities for inhibiting angiogenesis and tumor growth. PMID- 12086869 TI - Smart drugs: tyrosine kinase inhibitors in cancer therapy. AB - Cancer therapy directed at specific, frequently occurring molecular alterations in signaling pathways of cancer cells has been validated through the clinical development and regulatory approval of agents such as Herceptin for the treatment of advanced breast cancer and Gleevec for chronic myelogenous leukemia and gastrointestinal stromal tumors. While most novel, target-directed cancer drugs have pregenomic origins, one can anticipate a postgenomic wave of sophisticated "smart drugs" to fundamentally change the treatment of all cancers. With these prospects, interest in this new class of therapeutics extends from basic research scientists to practicing oncologists and their patients. An extension of the initial successes in molecular oncology will occur more quickly and successfully through an appreciation of lessons learned with the first group of agents in their progress through clinical development. PMID- 12086870 TI - Focus on central nervous system neoplasia. PMID- 12086871 TI - The BRCA1 suppressor hypothesis: an explanation for the tissue-specific tumor development in BRCA1 patients. PMID- 12086872 TI - Classification, subtype discovery, and prediction of outcome in pediatric acute lymphoblastic leukemia by gene expression profiling. AB - Treatment of pediatric acute lymphoblastic leukemia (ALL) is based on the concept of tailoring the intensity of therapy to a patient's risk of relapse. To determine whether gene expression profiling could enhance risk assignment, we used oligonucleotide microarrays to analyze the pattern of genes expressed in leukemic blasts from 360 pediatric ALL patients. Distinct expression profiles identified each of the prognostically important leukemia subtypes, including T ALL, E2A-PBX1, BCR-ABL, TEL-AML1, MLL rearrangement, and hyperdiploid >50 chromosomes. In addition, another ALL subgroup was identified based on its unique expression profile. Examination of the genes comprising the expression signatures provided important insights into the biology of these leukemia subgroups. Further, within some genetic subgroups, expression profiles identified those patients that would eventually fail therapy. Thus, the single platform of expression profiling should enhance the accurate risk stratification of pediatric ALL patients. PMID- 12086873 TI - ITF-2, a downstream target of the Wnt/TCF pathway, is activated in human cancers with beta-catenin defects and promotes neoplastic transformation. AB - In many cancers, inactivation of the adenomatous polyposis coli (APC) or Axin tumor suppressor proteins or activating mutations in beta-catenin lead to elevated beta-catenin levels, enhanced binding of beta-catenin to T cell factor (TCF) proteins, and increased expression of TCF-regulated genes. We found that the gene for the basic helix-loop-helix transcription factor ITF-2 (immunoglobulin transcription factor-2) was activated in rat E1A-immortalized RK3E cells following neoplastic transformation by beta-catenin or ligand-induced activation of a beta-catenin-estrogen receptor fusion protein. Human cancers with beta-catenin regulatory defects had elevated ITF-2 expression, and ITF-2 was repressed by restoring wild-type APC function or inhibiting TCF activity. Of note, ITF-2 promoted neoplastic transformation of RK3E cells. We propose that ITF 2 is a TCF-regulated gene, which functions in concert with other TCF target genes to promote growth and/or survival of cancer cells with defects in beta-catenin regulation. PMID- 12086874 TI - Astrocyte inactivation of the pRb pathway predisposes mice to malignant astrocytoma development that is accelerated by PTEN mutation. AB - We have inactivated pRb, p107, and p130 in astrocytes by transgenic expression of T(121) (a truncated SV40 T antigen) under the GFAP promoter. Founder mice died perinatally with extensive expansion of neural precursor and anaplastic astrocyte populations. In astrocytes, aberrant proliferation and extensive apoptosis were induced. Using a conditional allele of T(121), early lethality was circumvented, and adult mice developed high-grade astrocytoma, in which regions of decreased apoptosis expressed activated Akt. Indeed, astrocytoma development was accelerated in a PTEN(+/-), but not p53(+/-), background. These studies establish a highly penetrant preclinical model for astrocytoma based on events observed in the human disease and further provide insight into the role of PTEN mutation in astrocytoma progression. PMID- 12086875 TI - Bcl-2-dependent modulation of Ca(2+) homeostasis and store-operated channels in prostate cancer cells. AB - Antiapoptotic oncoprotein Bcl-2 has extramitochondrial actions due to its localization on the endoplasmic reticulum (ER); however, the specific mechanisms of such actions remain unclear. Here we show that Bcl-2 overexpression in LNCaP prostate cancer epithelial cells results in downregulation of store-operated Ca(2+) current by decreasing the number of functional channels and inhibiting ER Ca(2+) uptake through a reduction in the expression of calreticulin and SERCA2b, two key proteins controlling ER Ca(2+) content. Furthermore, we demonstrate that Ca(2+) store depletion by itself is not sufficient to induce apoptosis in Bcl-2 overexpressing cells, and that sustained Ca(2+) entry via activated store operated channels (SOCs) is required as well. Our data therefore suggest the pivotal role of SOCs in apoptosis and cancer progression. PMID- 12086876 TI - Small GTPases and tyrosine kinases coregulate a molecular switch in the phosphoinositide 3-kinase regulatory subunit. AB - Phosphoinositide 3-kinase (PI3K) type IA is a heterodimer of a catalytic subunit, p110, and a regulatory subunit, p85. Here we show that p85 contains a GTPase responsive domain and an inhibitory domain, which together form a molecular switch that regulates PI3K. H-Ras and Rac1 activate PI3K by targeting the GTPase responsive domain. The stimulatory effect of these molecules, however, is blocked by the inhibitory domain, which functions by binding to tyrosine-phosphorylated molecules and is neutralized by tyrosine phosphorylation. The complementary effects of tyrosine kinases and small GTPases on the p85 molecular switch result in synergy between these two classes of molecules toward the activation of the PI3K/Akt pathway. PMID- 12086877 TI - VEGF-A has a critical, nonredundant role in angiogenic switching and pancreatic beta cell carcinogenesis. AB - In the RIP1-Tag2 mouse model of pancreatic islet carcinoma, angiogenesis is switched on in a discrete premalignant stage of tumor development, persisting thereafter. Signaling through VEGF receptor tyrosine kinases is a well established component of angiogenic regulation. We show that five VEGF ligand genes are expressed in normal islets and throughout islet tumorigenesis. To begin dissecting their contributions, we produced an islet beta cell specific knockout of VEGF-A, resulting in islets with reduced vascularity but largely normal physiology. In RIP1-Tag2 mice wherein most oncogene-expressing cells had deleted the VEGF-A gene, both angiogenic switching and tumor growth were severely disrupted, as was the neovasculature. Thus, VEGF-A is crucial for angiogenesis in a prototypical model of carcinogenesis, whose loss is not readily compensated. PMID- 12086880 TI - The fabric of cancer cell biology-Weaving together the strands. PMID- 12086878 TI - Gene expression correlates of clinical prostate cancer behavior. AB - Prostate tumors are among the most heterogeneous of cancers, both histologically and clinically. Microarray expression analysis was used to determine whether global biological differences underlie common pathological features of prostate cancer and to identify genes that might anticipate the clinical behavior of this disease. While no expression correlates of age, serum prostate specific antigen (PSA), and measures of local invasion were found, a set of genes was identified that strongly correlated with the state of tumor differentiation as measured by Gleason score. Moreover, a model using gene expression data alone accurately predicted patient outcome following prostatectomy. These results support the notion that the clinical behavior of prostate cancer is linked to underlying gene expression differences that are detectable at the time of diagnosis. PMID- 12086881 TI - Of mice and women. AB - Transgenic mouse model faithfully reproduces human ovarian carcinoma and offers new opportunities for understanding the natural history of this frequently fatal disease. PMID- 12086882 TI - Disabling Abl-perspectives on Abl kinase regulation and cancer therapeutics. AB - Pharmacologic inhibition of the Bcr-Abl tyrosine kinase in human chronic myeloid leukemia leads to dramatic clinical responses, but relapses occur in advanced stage patients. New findings about Abl kinase domain regulation provide insight into novel strategies for targeted therapy. PMID- 12086883 TI - Functional genomics and the breast cancer problem. AB - The clinical treatment of primary breast cancers has been greatly complicated by the inability to accurately predict which tumors will eventually become invasive and metastatic and which will remain localized and indolent. Lacking the ability to discriminate between these two classes of breast cancer patients, oncologists often apply aggressive adjuvant therapy to women in both groups. However, the use of functional genomics analysis has now made it possible to assemble a set of gene markers, the expression of which enables one to predict, with reasonably high accuracy, whether or not the patient will relapse or remain tumor-free five years after initial diagnosis and treatment. PMID- 12086884 TI - A matter of life and death. AB - We propose that deregulation of proliferation, together with a reduction in apoptosis, creates a platform that is both necessary and can be sufficient for cancer. The secondary traits of diverse neoplasms are a consequence of cell proliferation, tissue expansion, and other outcomes of this platform. PMID- 12086885 TI - Perspectives on the development of a molecularly targeted agent. AB - STI571 (Gleevec, imatinib mesylate) exemplifies the successful development of a rationally designed, molecularly targeted therapy for the treatment of a specific cancer. This article reviews the identification of Bcr-Abl as a therapeutic target in chronic myelogenous leukemia and the steps in the development of an agent to specifically inactivate this abnormality. Issues related to clinical trials of molecularly targeted agents are discussed, including dose and patient selection, as are possible mechanisms of resistance to STI571. Lastly, the potential use of STI571 in other malignancies and the translation of this paradigm to other malignancies is explored. PMID- 12086886 TI - Cancer genomics. AB - The draft human genome sequence and the dissemination of high throughput technology provides opportunities for systematic analysis of cancer cells. Genome wide mutation screens, high resolution analysis of chromosomal abberations and expression profiling all give comprehensive views of genetic alterations in cancer cells. From these analyses will come a complete list of the genetic changes that drive malignant transformation and of the therapeutic targets that may be exploited for clinical benefit. PMID- 12086887 TI - Focus on lung cancer. PMID- 12086888 TI - Induction of ovarian cancer by defined multiple genetic changes in a mouse model system. AB - We have developed a mouse model for ovarian carcinoma by using an avian retroviral gene delivery technique for the introduction of multiple genes into somatic ovarian cells of adult mice. Ovarian cells from transgenic mice engineered to express the gene encoding the avian receptor TVA were efficiently infected in vitro with multiple vectors carrying coding sequences for oncogenes and marker genes. When target cells were derived from TVA transgenic mice deficient for p53, the addition of any two of the oncogenes c-myc, K-ras, and Akt were sufficient to induce ovarian tumor formation when infected cells were injected at subcutaneous, intraperitoneal, or ovarian sites. We demonstrated that the ovarian surface epithelium is the precursor tissue for these ovarian carcinomas, and that introduction of oncogenes causes phenotypic changes in the ovarian surface epithelial cells. The induced ovarian tumors in mice resembled human ovarian carcinomas in their rapid progression and intraperitoneal metastatic spread. PMID- 12086889 TI - Expression of a conditional AML1-ETO oncogene bypasses embryonic lethality and establishes a murine model of human t(8;21) acute myeloid leukemia. AB - The AML1/CBFbeta transcription factor complex, a frequent target of chromosomal translocations in leukemia, is essential for the generation of definitive hematopoietic stem cells. Paradoxically, expression of the acute myeloid leukemia associated AML1-ETO fusion protein in mice results not in leukemia, but in embryonic lethality due to an absence of normal hematopoiesis. To bypass the embryonic lethality, we generated a mouse strain with a conditional AML1-ETO knockin allele that contains a loxP bracketed transcriptional stop cassette 5' to the AML1-ETO fusion site. Activation of this allele in vivo by Cre-mediated recombination resulted in an enhanced replating efficiency of myeloid progenitors, but it did not block their differentiation, nor was it sufficient to induce leukemia. However, induction of cooperating mutations resulted in the development of an acute myeloid disease that mimicked many of the features of human AML1-ETO-expressing leukemia. PMID- 12086891 TI - A role for common fragile site induction in amplification of human oncogenes. AB - Oncogene amplification is an important process in human tumorigenesis, but its underlying mechanism is currently unknown. Cytogenetic analysis indicates that amplification of drug-selected genes in rodent cells is driven by recurrent breaks within chromosomal common fragile sites (CFSs), via the breakage-fusion bridge (BFB) mechanism. Here we show that BFB cycles drive the intrachromosomal amplification of the MET oncogene in a human gastric carcinoma. Our molecular evidence includes a "ladder-like" structure and inverted repeat organization of the MET amplicons. Furthermore, we show that the breakpoints, setting the centromeric amplicon boundaries, are within the CFS FRA7G region. Upon replication stress, this region showed perturbed chromatin organization, predisposing it to breakage. Thus, in vivo induction of CFSs can play an important role in human oncogenesis. PMID- 12086890 TI - Gene expression signatures define novel oncogenic pathways in T cell acute lymphoblastic leukemia. AB - Human T cell leukemias can arise from oncogenes activated by specific chromosomal translocations involving the T cell receptor genes. Here we show that five different T cell oncogenes (HOX11, TAL1, LYL1, LMO1, and LMO2) are often aberrantly expressed in the absence of chromosomal abnormalities. Using oligonucleotide microarrays, we identified several gene expression signatures that were indicative of leukemic arrest at specific stages of normal thymocyte development: LYL1+ signature (pro-T), HOX11+ (early cortical thymocyte), and TAL1+ (late cortical thymocyte). Hierarchical clustering analysis of gene expression signatures grouped samples according to their shared oncogenic pathways and identified HOX11L2 activation as a novel event in T cell leukemogenesis. These findings have clinical importance, since HOX11 activation is significantly associated with a favorable prognosis, while expression of TAL1, LYL1, or, surprisingly, HOX11L2 confers a much worse response to treatment. Our results illustrate the power of gene expression profiles to elucidate transformation pathways relevant to human leukemia. PMID- 12086892 TI - Placenta growth factor-1 antagonizes VEGF-induced angiogenesis and tumor growth by the formation of functionally inactive PlGF-1/VEGF heterodimers. AB - Tumor growth and metastasis require concomitant growth of new blood vessels, which are stimulated by angiogenic factors, including vascular endothelial growth factor (VEGF), secreted by most tumors. Whereas the angiogenic property and molecular mechanisms of VEGF have been well studied, the biological function of its related homolog, placenta growth factor (PlGF), is poorly understood. Here we demonstrate that PlGF-1, an alternatively spliced isoform of the PlGF gene, antagonizes VEGF-induced angiogenesis when both factors are coexpressed in murine fibrosarcoma cells. Overexpression of PlGF-1 in VEGF-producing tumor cells results in the formation of PlGF-1/VEGF heterodimers and depletion of the majority of mouse VEGF homodimers. The heterodimeric form of PlGF-1/VEGF lacks the ability to induce angiogenesis in vitro and in vivo. Similarly, PlGF-1/VEGF fails to activate the VEGFR-2-mediated signaling pathways. Further, PlGF-1 inhibits the growth of a murine fibrosarcoma by approximately 90% when PlGF-1 expressing tumor cells are implanted in syngeneic mice. In contrast, overexpression of human VEGF in murine tumor cells causes accelerated and exponential growth of primary fibrosarcomas and early hepatic metastases. Our data demonstrate that PlGF-1, a member of the VEGF family, acts as a natural antagonist of VEGF when both factors are synthesized in the same population of cells. The underlying mechanism is due to the formation of functionally inactive heterodimers. PMID- 12086893 TI - Introduction of Dr. Michael Sarr. PMID- 12086894 TI - The electronic environment: how has it, how will it, and how should it affect us? PMID- 12086895 TI - Pancreatic cancer: from genes to patient care. PMID- 12086896 TI - Duodenal carcinoid tumors: how aggressive should we be? AB - Duodenal carcinoid tumors are uncommon. It is not known whether they behave more like carcinoid tumors in the appendix (indolent course) or those in the ileum (often virulent)-crucial information for determining the need for radical resection. A retrospective review at our tertiary referral center (from 1976 to 1999) identified 27 patients with primary duodenal carcinoid lesions, excluding functional islet cell tumors. Endoscopic biopsy provided the diagnosis in 78% of patients. Treatment was by endoscopic excision (n = 11), transduodenal excision (n = 8), pancreaticoduodenectomy (n = 3), segmental distal duodenectomy (n = 2), or palliative operation (n = 2). One patient did not undergo operation because of comorbidity. Eighteen of 19 patients with tumors smaller than 2 cm remained disease free after local (endoscopic or transduodenal) excision. The exception was a patient with a small periampullary carcinoid lesion. In contrast, all four patients with carcinoid tumors 2 cm or larger who were resected for cure developed a recurrence (2 to 9 years postoperatively). We conclude that duodenal carcinoid tumors smaller than 2 cm may be excised locally; to ensure complete resection we recommend open transduodenal excision for tumors between 1 and 2 cm. Endoscopic follow-up is indicated. It is unclear whether patients with larger tumors benefit from more aggressive locoregional resection. Ampullary/periampullary carcinoid tumors should be considered separately, as their behavior is unpredictable. PMID- 12086897 TI - Analysis of tumor characteristics and survival in liver transplant recipients with incidentally diagnosed hepatocellular carcinoma. AB - The use of orthotopic liver transplantation (OLTX) for the treatment of hepatocellular carcinoma (HCC) has generally become restricted to carefully selected cases of small oligocentric tumors. However, it is not uncommon to find previously undetected HCC within recipient cirrhotic livers at the time of hepatectomy. The impact of unsuspected HCC on patient outcomes remains unclear. A retrospective analysis of our institutional experience with adult primary OLTX was performed comparing recipients with incidental HCC (group 1), recipients with known or suspected HCC (group 2), and recipients confirmed by pathologic examination to be tumor free (group 3). Between 1984 and 2000, 27 patients in group 1, 12 patients in group 2, and 612 patients in group 3 underwent primary OLTX. Tumors were smaller (P = 0.0172) in group 1 than in group 2; however, the number of tumors and the histologic findings were similar in the groups. Incidence of bilobar involvement, vascular invasion, portal vein tumor thrombus, lymphatic involvement, and distant metastasis at the time of OLTX did not differ significantly between these groups. Four-year patient survival appeared to be lower in group 1 (70.0%) than in group 3 (79.0%) (P = 0.0546); 4-year patient survival was significantly worse in group 2 (31.0%) compared to group 3 (P = 0.0106). Thus, in our experience, incidentally diagnosed cases of HCC possess many of the same features of malignancy as preoperatively diagnosed HCC. Indeed, patient survival after OLTX appears to be adversely affected by the presence of incidental HCC. PMID- 12086899 TI - Incidence of Helicobacter pylori in operatively managed acute nonvariceal upper gastrointestinal bleeding. AB - Helicobacter pylori is a known contributor to ulcerogenesis and nonvariceal acute upper gastrointestinal hemorrhage. Its incidence in operatively managed patients with upper gastrointestinal hemorrhage is ill defined. Patients undergoing surgery for upper gastrointestinal hemorrhage secondary to gastroduodenal ulceration between 1993 and 1998 at the University of Tennessee were retrospectively reviewed. Factors examined included age, nonsteroidal drug use, endoscopic intervention, urgency of operation, and H. pylori status confirmed by histologic examination. Forty-two patients had surgery with three excluded because of a lack of histologic evaluation. The site of bleeding was gastric in 23 and duodenal in 14. H. pylori infection was present in nine (39.1%) gastric and 11 (68.7%) duodenal ulcers. The incidence of H. pylori infection was reduced in those over 60 years of age (28.6%). Endoscopy was performed in all patients, but only two had biopsies for assessment of H. pylori. Operative morbidity was 17.9% and mortality was 5.1%. No patient had rebleeding following surgery. The incidence of H. pylori in this population is less than that reported in uncomplicated ulcer disease. Those older than 60 tended to be H. pylori negative. Endoscopic assessment for H. pylori was infrequent. Traditional indications for surgical intervention in ulcer hemorrhage should not be altered based on H. pylori status. PMID- 12086898 TI - PI-3' kinase and NF-kappaB cross-signaling in human pancreatic cancer cells. AB - Because tumor necrosis factor-alpha (TNF-alpha) and some chemotherapeutic agents activate both apoptosis and NF-kappaB-dependent antiapoptotic genes, they may neutralize their own antitumor effects. The cell-signaling mechanisms for such chemoresistance are not clear but may involve phosphotidylinositol-3' kinase (PI3K). To clarify this we examined whether cross-signaling between PI3K and NF kappaB enhances the antitumor effect of TNF-alpha in human pancreatic cancer cells. Quiescent pancreatic cancer cells (Panc-1, MiaPaCa-2) with TNF-alpha, Ly294002 (PI3K inhibitor), alone or combined, were restimulated with mitogen (10% fetal calf serum [FCS] to induce cell cycle entry). Proliferation (monotetrazolium), cell cycle progression (ApoBrDU and fluorescence-activated cell sorter analysis), and apoptosis (PARP cleavage; caspase-3 activation) were measured. Akt activation (Akt kinase assay) and IkappaBalpha degradation were determined by Western blot analysis. Translocation of NF-kappaB into the nucleus was examined by EMSA, whereas an NF-kappaB/luciferase reporter gene was used to quantify NF-kappaB-dependent gene expression. Statistical analysis was carried out by means of two-tailed t test (P <0.05). PI3K inhibition significantly enhanced the antiproliferative and proapoptotic effects of TNF-alpha in both cell lines, Ly294002 also blocked TNF-alpha-induced Akt activation but failed to alter cytoplasmic IkappaBalpha degradation or subsequent NF-kappaB nuclear translocation. NF-kappaB-dependent gene expression, however, was ultimately suppressed by Ly294002, suggesting that PI3k-dependent activation of NF-kappaB is IkappaBalpha independent. PI3K inhibition can block NF-kappaB-dependent gene expression regardless of cytoplasmic IkappaBalpha/NF-kappaB activation. Because it also regulates the antitumor effects of TNF-alpha, PI3K may in part determine NF-kappaB-induced chemoresistance in human pancreatic cancer. PMID- 12086900 TI - Esophagectomy for adenocarcinoma in patients 45 years of age and younger. AB - Esophageal adenocarcinoma in patients 45 years of age or younger is uncommon. We reviewed our experience with the surgical management of these patients to determine their clinical characteristics, pathologic findings, and treatment results. Thirty-two patients were identified through our surgical pathology database, and their medical records were reviewed to determine clinical characteristics, treatment, treatment-associated mortality, tumor staging, presence of Barrett's mucosa, and survival. In our series, patients were white (100%) males (96.9%) with a history of reflux (56.3%), cigarette smoking (40.6%), and alcohol consumption (59.4%), who presented with progressive solid food dysphagia (78.1%). A prior diagnosis of Barrett's mucosa or use of antireflux medications was noted in five patients each (15.6%). There were no operative deaths. Actuarial survival was 81.1% (95% confidence interval [CI] 66.1 to 96.2) at 12 months, 68.5% (95% CI 49.5 to 87.5) at 24 months, and 56.9% (95% CI 34.6 to 79.1) at 60 months. Our findings show that patients with esophageal adenocarcinoma 45 years of age or younger have similar clinical findings to those reported in other large series where the median age is in the sixth or seventh decade of life, supporting a uniform theory of tumor pathogenesis. Esophagectomy may be performed with low mortality, and survival is reasonable for early-stage disease. Young patients with Barrett's esophagus are not immune from the development of adenocarcinoma and need to be screened accordingly. PMID- 12086901 TI - Staging of pancreatic cancer before and after neoadjuvant chemoradiation. AB - Neoadjuvant chemoradiation therapy is used at many institutions for treatment of localized adenocarcinoma of the pancreas. Accurate staging before neoadjuvant therapy identifies patients with distant metastatic disease, and restaging after neoadjuvant therapy selects patients for laparotomy and attempted resection. The aims of this study were to (1) determine the utility of staging laparoscopy in candidates for neoadjuvant therapy and (2) evaluate the accuracy of restaging CT following chemoradiation. Staging laparoscopy was performed in 98 patients with radiographically potentially resectable (no evidence of arterial abutment or venous occlusion) or locally advanced (arterial abutment or venous occlusion) adenocarcinoma of the pancreas. Unsuspected distant metastasis was identified in 8 (18%) of 45 patients with potentially resectable tumors and 13 (24%) of 55 patients with locally advanced tumors by CT. Neoadjuvant chemoradiation therapy and restaging CT were completed in a total of 103 patients. Thirty-three patients with potentially resectable tumors by restaging CT underwent surgical exploration and resections were performed in 27 (82%). Eleven (22%) of 49 patients with locally advanced tumors by restaging CT were resected, with negative margins in 55%; the tumors in these 11 patients had been considered locally advanced because of arterial involvement on restaging CT. Staging laparoscopy is useful for the exclusion of patients with unsuspected metastatic disease from aggressive neoadjuvant chemoradiation protocols. Following neoadjuvant chemoradiation, restaging CT guides the selection of patients for laparotomy but may overestimate unresectability to a greater extent than does prechemoradiation CT. PMID- 12086902 TI - Pancreatic epithelial cyst in an adult treated by central pancreatectomy. AB - The infrequent occurrence of benign epithelial cysts of the pancreas is the reason why little is known regarding their clinical relevance and surgical management. We report the case of a 38-year-old woman with a benign epithelial cyst that was resected by the rarely performed central pancreatectomy. The presentation, evaluation, and differences between this and other cystic lesions of the pancreas are discussed. The benefits of central pancreatectomy for this benign lesion are reviewed. PMID- 12086903 TI - Multimodality treatment for patients with hepatocellular carcinoma: analysis of prognostic factors in a single Western institution series. AB - There are few Western studies evaluating prognostic factors for survival in patients with hepatocellular carcinoma (HCC) and the influence on survival of various therapeutic options including orthotopic liver transplantation (OLT). A retrospective analysis was performed of 122 patients with HCC treated at the University of Alabama at Birmingham from January 1990 through December 1999. Clinicopathologic and treatment factors were analyzed with overall survival as the main outcome variable. Median age was 62 years. Most patients were male (74%) and white (79%). Eighty patients (66%) had associated cirrhosis. Sixty-three percent of patients presented with American Joint Committee on Cancer (AJCC) stage III or IV tumors. The median follow-up for survivors was 22 months. The 1-, 3-, and 5-year actuarial survival rates for the entire cohort were 46%, 24%, and 17%, respectively. On multivariate analysis, ablative surgery (P = 0.003), AJCC stages I and II (P = 0.0012), and absence of vascular invasion (P = 0.0001) were found to be independent favorable characteristics. Forty-four patients underwent surgical resection (including OLT, n = 20) or a surgical ablative procedure. All but two nonsurgical patients died of disease. The actuarial 1-, 3-, and 5-year survival rates for this group were 80%, 71%, and 61%, respectively. On multivariate analysis of the surgical group, only vascular invasion was associated with poor prognosis (P = 0.001). OLT was associated with a favorable prognosis on univariate analysis (P = 0.02). Forty percent of patients who received transplants underwent local/regional treatment before transplantation and the outcome in these patients was no different from that in other transplant patients. Surgical treatment is the only potential curative option for HCC, and qualifying for liver transplantation may be a favorable prognostic factor in surgical patients. Local/regional therapy prior to transplantation may provide a bridge to OLT without an increase in tumor-related mortality. PMID- 12086904 TI - Effect of interstitial laser hyperthermia in a murine model of colorectal liver metastases. AB - Interstitial laser hyperthermia (ILH) is an in situ ablative technique used for the treatment of colorectal liver metastases. At present, few data exist concerning the optimum power settings required to maximize tissue necrosis. The aim of this study was to establish the dose-response relationship between the laser power setting and the extent of tissue necrosis produced in liver and tumor tissue, as well as the pattern of necrosis in a murine model of liver metastases. An intrasplenic induction model of liver metastases in 4- to 6-week-old male inbred CBA mice was used. Laser hyperthermia was applied to liver and tumor tissue using a bare optical quartz fiber from a Laserex SLY500 Nd:YAG surgical laser generator. Two-watt and 5-watt power settings were used at specific time intervals. The livers were then excised, fixed in formalin, and the extent and degree of necrosis were measured. Results were expressed as mean +/- standard deviation and were normally distributed. Analysis of variance was performed, and the least significant difference was used for post hoc tests. A P value of less than 0.05 was considered significant. Interstitial laser hyperthermia at 5 watts of power produced larger diameters of necrosis than did 2 watts for specific exposure times in normal liver tissue. However, when the total energy applied was compared, there was no significant difference in the diameters of tissue necrosis produced by the two power settings. The diameter of tissue necrosis in the normal liver increased from 2 mm at 10 joules to 8 mm at 600 joules of energy. Within tumor tissue, ILH at 2 and 5 watts produced similar diameters of necrosis for specific exposure times. When amounts of total energy applied were compared, ILH at the lower power setting (2 watts) produced a significantly larger diameter of necrosis than the higher power setting (5 watts). The diameter of necrosis achieved in tumor tissue was significantly larger than that in normal liver tissue at both power settings, for an equivalent amount of applied energy. The difference was more pronounced when ILH was performed at the lower power setting. The maximum diameter of necrosis achieved was 6.8 +/- 0.7 mm in normal liver tissue and 7.7 +/- 0.8 mm in tumor tissue. Charring of the fiber tip was delayed when the lower power setting was used, occurring after 20 seconds of exposure, compared to 5 seconds at the higher power setting. Similarly, cavitation occurred initially at 50 seconds at 5 watts of power and was delayed until 90 seconds of exposure at 2 watts of power. Histopathologic findings revealed an elliptical area of homogeneous necrosis, with a central acellular coagulum surrounded by intact but non-viable tissue. ILH is capable of producing highly reproducible, uniform, and complete tissue necrosis. The diameter of necrosis is related to the total energy applied. At low-power settings at any given amount of applied energy, a significantly larger diameter of tissue necrosis was achieved in tumor tissue compared to normal liver tissue. PMID- 12086905 TI - Concomitant sclerosing mesenteritis and bile duct fibrosis simulating Klatskin's tumor. AB - Sclerosing mesenteritis is an uncommon benign condition that should be included in the differential diagnosis of abdominal masses. We present the first reported case of this condition in association with idiopathic bile duct fibrosis simulating Klatskin's tumor. A review of the literature regarding both clinical entities is presented. PMID- 12086906 TI - Cholinergic stimulation of rat acinar cells increases c-fos and c-jun expression via a mitogen-activated protein kinase-dependent pathway. AB - Acetylcholine release from cholinergic neurons regulates pancreatic exocrine function through pathways that are still under investigation. Pancreatic AR42J acinar cells were studied to determine intracellular calcium ([Ca(2+)](i)) release, enzyme activation, and gene expression in response to the acetylcholine analog carbachol (CCh). CCh stimulated dose-dependent increases in [Ca(2+)](i) that were inhibited by atropine and by specific inhibitors to the muscarinic receptor subtypes m1 and m3. Polymerase chain reaction analysis was performed, which sequenced products corresponding to the m1 and m3 receptor subtypes but not the m2 subtype. CCh also stimulated mitogen-activated protein kinase activity. CCh induced time-and dose-dependent increases in the c-fos and c-jun early response genes, which were blocked by m1 and m3 inhibition but not by m2 inhibition. PMID- 12086909 TI - Resected adenocarcinoma of the pancreas--616 patients: results, outcomes, and prognostic indicators. PMID- 12086907 TI - Expression of 17-1A antigen and complement resistance factors CD55 and CD59 on liver metastasis in colorectal cancer. AB - Despite radical surgery, the prognosis for colorectal cancer patients with liver metastases has not changed markedly. Furthermore, no standard adjuvant therapeutic regimen has been developed. Adjuvant therapy with monoclonal antibodies (e.g., against 17-1A), which has been shown to be effective in preventing metastatic relapse in patients with Dukes' C colorectal cancer, might be a promising approach for these patients. However, the cytotoxic effects of monoclonal antibodies can be blocked by coexpression of complement resistance factors that inhibit antibody-dependent complement-mediated cytotoxicity. We therefore analyzed immunohistochemically the expression of 17-1A and the membrane bound complement resistance factors CD55 and CD59 on metastatic tumor cells in the livers of 71 patients with colorectal carcinoma who had undergone resection of their metastases with curative intent. In 67 (94%) of 71 patients, liver metastases with homogeneous expression of 17-1A was seen. Heterogeneous expression of 17-1A was seen in four patients (6%). Heterogeneous expression of CD55 or CD59 was observed in 8 (11%) of 71 patients and 4 (6%) of 71 patients, respectively. None of the patients showed homogeneous expression of either CD55 or CD59. All patients with CD55 or CD59 expression showed homogeneous 17-1A expression, whereas none of the four patients with heterogeneous 17-1A expression was positive for CD55 or CD59. Our data indicate that 17-1A is widely expressed on liver metastases of patients with colorectal carcinoma. Therefore patients with completely resected liver metastases might be suitable candidates for adjuvant therapy with and-17-1A antibody since only a few of these lesions showed coexpression of complement resistance factors. PMID- 12086911 TI - Molecular pathogenesis of T lymphocyte-induced liver injury in alcoholic hepatitis. AB - The development of alcohol-induced liver injury is, in part, a consequence of the immunological/inflammatory response that alcohol stimulates. The abnormalities of immune function in heavy drinkers have been documented well. Cytokines, especially TNF alpha, produced from macrophages/Kupffer cells, play a role in the induction of liver cell necrosis and apoptosis. TNF alpha can cause liver cell apoptosis through the TNF alpha receptor or Fas/CD95 which is expressed by liver cells. Furthermore, chronic ethanol consumption may damage the liver by inhibiting the hepatotrophic and hepatoprotective actions of TNF alpha and other cytokines. There exists an intrinsic lymphocyte population in the normal liver. Intrahepatic T lymphocytes consist of a heterogeneous population of cells that has many and varied functional characteristics in addition to classical T cell activity. The population of intrahepatic T lymphocytes may arise via a thymus independent pathway. Our recent work has demonstrated the role of liver associated T lymphocytes in the pathogenesis of alcohol related liver injury initiated by a variety of stimuli such as endotoxin (lipopolysaccharide, LPS) or concanavalin A (Con A). Our studies have, for the first time, suggested that alcohol consumption alone does not lead to the development of marked liver necrosis (at least in the rat), but rather that a second insult is required for this to occur. Liver-associated T lymphocytes in rats spontaneously secrete interleukin-1 alpha, interleukin-6 and TNF alpha in vitro culture. There is a significant decline in the amounts of interleukin-1 alpha and TNF alpha secreted in ethanol-consuming rats compared with non-ethanol consuming rats. The numbers of T cells, NK cells and Kupffer cells in liver perfusates remains stable over a prolonged period of ethanol consumption. However, following Con A injection, there was an inappropriate increase in the amounts of interleukin-6 and TNF alpha secreted in in vitro culture of liver-associated T lymphocytes and a significant increase in the percentage of CD4+ T cells and CD25+ T cells in liver perfusates compared with non-ethanol consuming rats. It suggested that liver-associated T lymphocytes are involved in the inflammatory process associated with alcohol related liver injury through increased cytokine secretion (TNF alpha). PMID- 12086912 TI - Mechanisms of alcohol-mediated CD4+ T lymphocyte death: relevance to HIV and HCV pathogenesis. AB - Clinical and experimental studies have demonstrated that excessive alcohol consumption can result in impairment of the immune system, and can impact several immune functions including immune tolerance and host defense against opportunistic infections, and development of certain tumors. Although multiple factors are involved in the effects of ethanol on the immune system, several studies implicate chronic activation of immune cells and impairment of thymus derived lymphocytes (T lymphocytes). Helper CD4+ T lymphocytes are the central regulators of the immune system and depletion of these lymphocytes is a major contributing factor in ethanol-induced immune dysfunction and exacerbation of HIV and/or HCV pathogenesis. However, the mechanisms involved in the ethanol induced CD4+ T cell depletion have only recently begun to be elucidated. Our work demonstrates that exposure of human CD4+ T cells to physiologically relevant concentrations of ethanol leads to the (i) enhanced activation of TNFalpha inducible NFkappaB, the transcriptional regulator of the Fas promoter and ii) increased susceptibility to Fas-and activation-induced apoptotic death via augmentation of caspase 3 activity. Work done by us, and others, on the effects of ethanol on CD4+ T cell function and survival strongly suggests that alcohol plays a significant role as a co-factor in HIV and/or HCV pathogenesis. PMID- 12086913 TI - Cellular scaffolds in mammalian eggs. AB - Cellular scaffolds serve as structural components to which various elements of signal transduction pathways can be associated. The association of components on a scaffold can have several important functions, for example they can: 1) associate upstream regulatory components in a cascade that can increase the speed of response to a stimulus; 2) restrict access of substrates to enzymes associated with the scaffold; 3) permit cross talk between distinct signaling pathways, and; 4) aid in the establishment of cellular polarity. The conversion of the mammalian egg into the zygote requires many rapid alterations during a distinct time frame to mediate the biochemical and structural changes that occur. Cellular scaffolds provide a mechanism that can perform these rapid, highly orchestrated changes. They can permit interaction between distinct calcium-dependent pathways and also can provide a means for the calcium signal, that is initiated by fertilization, to act on calcium-independent pathways. This review considers various lines of evidence suggesting that in the mammalian egg, the meiotic spindle serves as a cellular scaffold that permits coordination among several signaling pathways essential for fertilization and the initiation of early development. PMID- 12086914 TI - The role of vascular growth factors in hyperoxia-induced injury to the developing lung. AB - Normal pulmonary vascular development is the result of a complex interplay of growth factors, including vascular endothelial growth factor (VEGF) and the angiopoietins. Injury to the developing lung, whether due to hyperoxia or mechanical ventilation, results in disordered vascular development, ranging from an apparent arrest of microvascular development in milder injury to extensive microvascular derangement in more severe injury. Alterations in vascular growth factors may participate in these injuries. During injury to the developing animal lung, VEGF abundance is markedly decreased. In models of post-injury recovery, up regulation of VEGF accompanies the re-establishment of normal vasculature. Alterations in lung VEGF levels in human premature infants are less clear cut. However, among humans premature newborns who later go on to develop bronchopulmonary dysplasia (BPD), VEGF production is decreased in comparison to those newborns who recover. Other angiogenic factors, such as the CXC ELR+ chemokines, are also altered in injury to the developing lung, but their specific roles in vascular injury are less clear. Strategies that enhance microvascular integrity, whether through attenuating alterations in vascular growth factors or by other means, also improve the outcome of lung injury. Such therapies may eventually offer hope in human BPD. PMID- 12086915 TI - The role of cholangiocytes in the development of chronic inflammatory liver disease. AB - Cholangiocytes constitute the biliary epithelium, an important barrier to infection entering the liver via the gastrointestinal tract. These cells have developed mechanisms to respond to infection by recruiting and interacting with effector leukocytes to clear bacterial or viral pathogens. Cholangiocytes are also targets of immune mediated damage in several liver diseases and under these circumstances protective mechanisms, for example the secretion of chemokines and expression of adhesion molecules, become harmful and promote the inappropriate recruitment and retention of effector cells within portal tracts. In chronic inflammatory biliary diseases such as primary biliary cirrhosis and primary sclerosing cholangitis, infiltrating leukocytes destroy bile ducts by killing cholangiocytes via complex molecular mechanisms involving Fas-dependent apoptosis and autocrine and paracrine interactions with other members of the TNF superfamily including CD40. A better understanding of these processes may lead to novel therapeutic approaches aimed at switching off the chronic inflammatory response and protecting bile ducts from apoptosis. PMID- 12086916 TI - Intracellular Ca2+ store in embryonic cardiac myocytes. AB - In mature cardiac myocytes, Ca2+ influx through the L-type Ca2+ channel activates the ryanodine receptor and triggers Ca2+ release from the sarcoplasmic reticulum (SR). This Ca2+ signal amplification, termed Ca2+-induced Ca2+ release (CICR), occurs within the junctional membrane complex between the plasma membrane and the SR, and is essential for cardiac excitation-contraction (E-C) coupling. On the other hand, Ca2+ available during E-C coupling is predominantly derived from Ca2+ influx in embryonic cardiac myocytes. To examine the role of the intracellular Ca2+ store in immature cardiac myocytes, we have generated knockout mice lacking the cardiac type of the ryanodine receptor (RyR-2), or junctophilin (JP-2) contributing to formation of the junctional membrane complex. Both RyR-2- and JP 2-knockout mice show lethality at early embryonic stages immediately after beginning of heart beating. The loss of RyR-2 produced abnormal SR elements exhibiting vacuolated structures and Ca2+-overloading in embryonic cardiac myocytes. In JP-2-deficient cardiac myocytes, formation of junctional membrane complexes, called pheripheral couplings, was disturbed, and abnormal Ca2+ transients without spatial and temporal synchronization were observed. Therefore, the knockout mice have demonstrated that RyR-2-mediated Ca2+ release at the junctional membrane complex is essential for cellular Ca2+ homeostasis in immature cardiac myocytes. PMID- 12086917 TI - The role of biliary calcium in gallstone pathogenesis. AB - Calcium is an element that is extremely important in the pathogenesis of gallstones. It is present in bile in large quantities since components of bile bind the calcium avidly. Precipitated calcium salts with the calcium sensitive ions, bilirubinate, carbonate and phosphate are major components of pigment gallstones and are present in high concentrations in the centers and rims of cholesterol gallstones. An understanding of the solubility of calcium in bile is thus essential if one is to understand the process that leads to gallstone formation. In addition, calcium also has a very important role in the function of the gallbladder epithelial cell. This manuscript discusses what is known about biliary calcium, the factors that regulate total and free calcium concentrations in bile, the solubility of calcium in both normal and lithogenic bile, and the role calcium plays in epithelial cell function during gallstone formation. PMID- 12086918 TI - Hepatitis C virus (HCV) and human immunodeficiency virus type 1 (HIV-1) infections in alcoholics. AB - Approximately 400,000 individuals in the United States are co-infected with hepatitis C virus (HCV) and human immunodeficiency virus type 1 (HIV-1) and it is likely that almost one in two of these subjects consumes alcohol. The majority of these patients suffer an accelerated course of liver disease as manifested by the onset of cirrhosis within 5 to 10 years of developing HCV infection, as well as an increased risk of developing hepatocellular carcinoma (HCC). It is thought that chronic alcohol abuse mediates liver damage as a result of increased production of free radicals and proinflammatory cytokines. In the setting of chronic HCV infection, alcohol ingestion has an additional effect of diminishing immune clearance and increasing viral burden to hasten the onset of cirrhosis and HCC. Likewise, chronic HCV and HIV-1 co-infection results in a net increase in HCV burden; higher prevalence rates of HCV transmission to sexual partners and offspring, as well as an accelerated progression to end stage liver disease as compared to individuals with HCV infection alone. Thus, the synergistic effects of alcohol abuse and HIV-1 greatly impact on the morbidity and mortality for patients with HCV coinfection. Ultimately, this cumulative disease process will require far more aggressive management with abstinence and counseling for alcohol abuse; highly active antiretroviral therapy (HAART) for HIV infection and combination anti-viral therapy for HCV infection to stem the rapid progression to end stage liver disease. PMID- 12086919 TI - Theoretical approach to endocrine disruptors. AB - Endocrine disruptors are now of scientific and public concern, because there is increasing evidence of their adverse effects on the health of an intact organism or its progeny and on changes in endocrine function. Although numerous substances have been identified as such chemicals, a huge number of chemicals remain to be tested for their endocrine disrupting capabilities. Because of the time and costs required for animal tests, some theoretical or computer-based method for screening this large number of chemicals is needed to reduce the numbers requiring animal testing. Improved quantitative structure activity relationship (QSAR) models were used for screening in combination with other approaches. New receptor-ligand docking simulations were being tested. There was good correlation between experimental and theoretical binding affinities. A database complex system being developed, which enables one to trace cellular signals triggered by the interaction of receptors with xenobiotic chemicals. Perspectives of computer based screening methods are discussed. PMID- 12086920 TI - The effect of phospholipid transfer protein on lipoprotein metabolism and atherosclerosis. AB - Phospholipid transfer protein (PLTP) is a member of the lipid transfer/lipopolysaccharide binding protein gene family. Recently, the crystal structure of one of the members of the gene family, bactericidal permeability increasing protein, was solved, providing potential insights into the mechanisms of action of PLTP. These molecules contain intrinsic lipid binding sites and appear to act as carrier proteins that shuttle between lipoproteins to redistribute lipids. The phenotype of PLTP transgenic and gene knock out mice indicates that PLTP plays a major role in the metabolism of high-density lipoprotein (HDL) and apoB-containing lipoproteins and thereby influences the concentration, apolipoprotein content, and size of lipoprotein particles in plasma. Recent data indicate that PLTP deficiency in mice is associated with a decrease of atherosclerosis, despite decreased HDL levels. At lease two underlined mechanisms are involved in the reduction of atherosclerosis in PLTP deficient status, 1) reduction of apoB-containing lipoprotein production and levels; and 2) increase of anti-oxidation potential. Human studies indicated that PLTP activity positively correlated with aging, obesity, diabetes and coronary artery disease. A challenge for the future will be the inhibition of PLTP for therapeutic benefit. PMID- 12086921 TI - Ryanodine receptors in smooth muscle. AB - The sarcoplasmic reticulum (SR) of smooth muscle is endowed with two different types of Ca2+ release channels, i.e. inositol 1,4,5-trisphosphate receptors (IP3Rs) and ryanodine receptors (RyRs). In general, both release channels mobilize Ca2+ from the same internal store in smooth muscle. While the importance of IP3Rs in agonist-induced contraction is well established, the role of RyRs in excitation-contraction coupling of smooth muscle is not clear. The participation of smooth muscle RyRs in the amplification of Ca2+ transients induced by either opening of Ca2+-permeable channels or IP3-triggered Ca2+ release has been studied. The efficacy of both processes to activate RyRs by calcium-induced calcium release (CICR) is highly variable and not widely present in smooth muscle. Although RyRs in smooth muscle generate Ca2+ sparks that are similar to those observed in striated muscles, the contribution of these local Ca2+ events to depolarization-induced global rise in [Ca2+]i is rather limited. Recent data suggest that RyRs are involved in regulating the luminal [Ca2+] of SR and also in smooth muscle relaxation. This review summarizes studies that were carried out mainly in muscle strips or in freshly isolated myocytes, and that were aimed to determine the physiological role of RyRs in smooth muscle. PMID- 12086922 TI - Liver regeneration. AB - Unlike other vital organs, the liver typically regenerates after injury. Indeed, the very factors that cause liver injury initiate a reparative process in the residual liver that includes the induction of cytoprotective mechanisms, deletion of mortally wounded cells, repair of less damaged survivors, liver cell proliferation to replace the cells that died, the deposition of new matrix, and tissue remodeling to restore normal hepatic mass and architecture. During liver regeneration, the liver normally continues to perform vital, liver-specific functions. Unfortunately, the hepatic regenerative response sometimes becomes disrupted--either failing to occur or occurring in a disordered, or incomplete fashion. Abnormal regeneration contributes to the pathogenesis of fulminate liver failure, cirrhosis, and primary liver cancers. Research in the field of regenerative biology has identified several events that are required for liver regeneration. These include injury-induced changes in the hepatic microenvironment, the ability of surviving liver cells and/or their progenitors to proliferate, and a temporary suspension of homeostatic mechanisms that normally couple cell proliferation to programmed cell death. The signals that mediate these complex biologic responses are being detailed. A better understanding of the extra- and intracellular signals that prompt the injured liver to regenerate should suggest treatments to promote liver regeneration in patients with otherwise fatal liver diseases. PMID- 12086924 TI - Sarcoplasmic reticulum Ca release in intact ventricular myocytes. AB - Sarcoplasmic reticulum (SR) Ca release in intact ventricular myocytes is the major source of Ca which activates cardiac contraction (although Ca influx makes a non-negligible contribution in most species). The fundamental events of SR Ca release are known as Ca sparks. The twitch Ca transient is composed of approximately 10,000 Ca sparks occurring in a given cell, and they are synchronized by the action potential and Ca current. Many factors influence SR Ca release amplitude and kinetics, and the focus here is on understanding how these factors work in the intact cellular environment. The intracellular [Ca] ([Ca]i) and intra-SR [Ca] ([Ca]SR) are two of the most important dynamic modulators of SR Ca release. Indeed, while [Ca]i (and Ca current which initiates systolic SR Ca release) is widely acknowledged to be important, it is increasingly clear that [Ca]SR changes dynamically under physiological conditions and that this has very important regulatory effects on the SR Ca release process. While elevation of [Ca]SR obviously increases the driving force and amount of SR Ca available for release, it also increases the fractional release and can be responsible for spontaneous diastolic SR Ca release. These issues are discussed in both normal physiological and pathophysiological contexts. PMID- 12086925 TI - Functional genomics of the endocrine pancreas: the pancreas clone set and PancChip, new resources for diabetes research. AB - Over the past 5 years, microarrays have greatly facilitated large-scale analysis of gene expression levels. Although these arrays were not specifically geared to represent tissues and pathways known to be affected by diabetes, they have been used in both type 1 and type 2 diabetes research. To prepare a tool that is particularly useful in the study of type 1 diabetes, we have assembled a nonredundant set of 3,400 clones representing genes expressed in the mouse pancreas or pathways known to be affected by diabetes. We have demonstrated the usefulness of this clone set by preparing a cDNA glass microarray, the PancChip, and using it to analyze pancreatic gene expression from embryonic day 14.5 through adulthood in mice. The clone set and corresponding array are useful resources for diabetes research. PMID- 12086926 TI - Lipid-induced insulin resistance in human muscle is associated with changes in diacylglycerol, protein kinase C, and IkappaB-alpha. AB - The possibility that lipid-induced insulin resistance in human muscle is related to alterations in diacylglycerol (DAG)/protein kinase C (PKC) signaling was investigated in normal volunteers during euglycemic-hyperinsulinemic clamping in which plasma free fatty acid (FFA) levels were increased by a lipid/heparin infusion. In keeping with previous reports, rates of insulin-stimulated glucose disappearance (G(Rd)) were normal after 2 h but were reduced by 43% (from 52.7 +/ 8.2 to 30.0 +/- 5.3 micromol. kg(-1). min(-1), P < 0.05) after 6 h of lipid infusion. No changes in PKC activity or DAG mass were seen in muscle biopsy samples after 2 h of lipid infusion; however, at approximately 6 h, PKC activity and DAG mass were increased approximately fourfold, as were the abundance of membrane-associated PKC-betaII and -delta. A threefold increase in membrane associated PKC-betaII was also observed at approximately 2 h but was not statistically significant (P = 0.058). Ceramide mass was not changed at either time point. To evaluate whether the fatty acid-induced insulin activation of PKC was associated with a change in the IkB kinase (IKK)/nuclear factor (NF)-kappaB pathway, we determined the abundance in muscle of IkappaB-alpha, an inhibitor of NF-kappaB that is degraded after its phosphorylation by IKK. In parallel with the changes in DAG/PKC, no change in IkappaB-alpha mass was observed after 2 h of lipid infusion, but at approximately 6 h, IkappaB-alpha was diminished by 70%. In summary, the results indicated that the insulin resistance observed in human muscle when plasma FFA levels were elevated during euglycemic-hyperinsulinemic clamping was associated with increases in DAG mass and membrane-associated PKC betaII and -delta and a decrease in IkappaB-alpha. Whether acute FFA-induced insulin resistance in human skeletal muscle is caused by the activation of these specific PKC isoforms and the IKK-beta/IkappaB/NFkappaB pathway remains to be established. PMID- 12086927 TI - The gene INPPL1, encoding the lipid phosphatase SHIP2, is a candidate for type 2 diabetes in rat and man. AB - Genetic susceptibility to type 2 diabetes involves many genes, most of which are still unknown. The lipid phosphatase SHIP2 is a potent negative regulator of insulin signaling and sensitivity in vivo and is thus a good candidate gene. Here we report the presence of SHIP2 gene mutations associated with type 2 diabetes in rats and humans. The R1142C mutation specifically identified in Goto-Kakizaki (GK) and spontaneously hypertensive rat strains disrupts a potential class II ligand for Src homology (SH)-3 domain and slightly impairs insulin signaling in cell culture. In humans, a deletion identified in the SHIP2 3' untranslated region (UTR) of type 2 diabetic subjects includes a motif implicated in the control of protein synthesis. In cell culture, the deletion results in reporter messenger RNA and protein overexpression. Finally, genotyping of a cohort of type 2 diabetic and control subjects showed a significant association between the deletion and type 2 diabetes. Altogether, our results show that mutations in the SHIP2 gene contribute to the genetic susceptibility to type 2 diabetes in rats and humans. PMID- 12086928 TI - Critical role for cataplerosis via citrate in glucose-regulated insulin release. AB - The molecular mechanisms mediating acute regulation of insulin release by glucose are partially known. The process involves at least two pathways that can be discriminated on basis of their (in)dependence of closure of ATP-sensitive potassium (K+(ATP)) channels. The mechanism of the K+(ATP) channel-independent pathway was proposed to involve cataplerosis, the export of mitochondrial intermediates into the cytosol and in the induction of fatty acid-derived signaling molecules. In the present article, we have explored in fluorescence activated cell sorter (FACS)-purified rat beta-cells the molecular steps involved in chronic glucose regulation of the insulin secretory response. When compared with culture in 10 mmol/l glucose, 24 h culture in 3 mmol/l glucose shifts the phenotype of the cells into a state with low further secretory responsiveness to glucose, lower rates of glucose oxidation, and lower rates of cataplerosis. Microarray mRNA analysis indicates that this shift can be attributed to differences in expression of genes involved in the K+(ATP) channel-dependent pathway, in cataplerosis and in fatty acid/cholesterol biosynthesis. This response was paralleled by glucose upregulation of the transcription factor sterol regulatory element binding protein 1c (SREBP1c) (ADD1) and downregulation of peroxisome proliferator-activated receptor (PPAR)-alpha and PPAR-beta (PPARdelta). The functional importance of cataplerosis via citrate for glucose induced insulin release was further supported by the observation that two ATP citrate lyase inhibitors, radicicol and (-)-hydroxycitrate, block part of glucose stimulated release in beta-cells. In conclusion, chronic glucose regulation of the glucose-responsive secretory phenotype is associated with coordinated changes in gene expression involved in the K+(ATP) channel-dependent pathway, in cataplerosis via citrate and in acyl CoA/cholesterol biosynthesis. PMID- 12086929 TI - Renal perfusion and the renal hemodynamic response to blocking the renin system in diabetes: are the forces leading to vasodilation and vasoconstriction linked? AB - In three groups of subjects, those with type 2 diabetes and nephropathy, those with type 1 diabetes without nephropathy, and healthy volunteers subjected to short-term hyperglycemia, we observed a counterintuitive relationship. In all three groups, baseline renal plasma flow (RPF) was positively correlated with the RPF response to blocking the renin-angiotensin system (RAS). This seems paradoxical in that an opposite result would have been expected if angiotensin dependent renal vasoconstriction was responsible for the renal vasodilator response to RAS blockade. This suggests a link between the renal vasodilator response, mediated by nitric oxide (NO), and the activation of the intrarenal RAS. The complex interrelationships between hyperglycemia, insulin, NO, and the RAS may result in phenotypes that indicate varying risk of diabetic nephropathy and underlying genetic polymorphisms. PMID- 12086930 TI - Increased lipolysis and decreased leptin production by human omental as compared with subcutaneous preadipocytes. AB - Site differences in adipose tissue function may have implications for insulin resistant conditions. In mature adipose tissue, subcutaneous adipocytes have higher leptin secretion, similar tumor necrosis factor (TNF)-alpha secretion, and lower catecholamine-stimulated lipolysis as compared with omental adipocytes. In this study, lipolysis and leptin and TNF-alpha secretion were compared between human omental and subcutaneous preadipocytes. After 16 days of incubation in a minimal differentiation medium, leptin mRNA and secretion were found to be two to eight times higher in subcutaneous than omental preadipocytes (P < 0.05). On the other hand, norepinephrine-induced lipolysis was about two times higher in the omental than in the subcutaneous preadipocytes, whereas basal lipolysis did not differ between the two regions. TNF-alpha secretion was marginally but significantly higher in the omental than in the subcutaneous preadipocytes. Preadipocyte differentiation was equal in both regions and was augmented to the same extent by different thiazolidinediones (rosiglitazone, pioglitazone, or troglitazone) in the two depots. In the presence of rosiglitazone, leptin secretion remained about three times higher and norepinephrine-induced lipolysis about six times lower in subcutaneous as compared with omental preadipocytes (P < 0.05), whereas TNF-alpha secretion and basal lipolysis were similar in preadipocytes from the two regions. These findings remained unaltered even if rosiglitazone was removed from the medium. However, leptin mRNA showed no regional differences in rosiglitazone-treated cells. Thus, regional differences in adipocyte leptin secretion as well as in norepinephrine-induced lipolysis are marked and present during different stages of preadipocyte differentiation and seem to be determined by intrinsic (i.e., primary) factors. PMID- 12086931 TI - Isomer-dependent metabolic effects of conjugated linoleic acid: insights from molecular markers sterol regulatory element-binding protein-1c and LXRalpha. AB - Conjugated linoleic acid (CLA) is a heterogeneous group of positional and geometric isomers of linoleic acid. This study demonstrates the divergent effects of the cis-9 trans-11 (c9,t11-CLA) and trans-10 cis-12 (t10,c12-CLA) isomers of CLA on lipid metabolism and nutrient regulation of gene expression in ob/ob mice. The c9, t11-CLA diet decreased serum triacylglycerol (P = 0.01) and nonesterified fatty acid (NEFA) (P = 0.05) concentrations, and this was associated with reduced hepatic sterol regulatory element-binding protein-1c (SREBP-1c; P = 0.0045) mRNA expression, coupled with reduced levels of both the membrane-bound precursor and the nuclear forms of the SREBP-1 protein. C9,t11-CLA significantly reduced hepatic LXRalpha (P = 0.019) mRNA expression, a novel regulator of SREBP-1c. In contrast, c9,t11-CLA increased adipose tissue SREBP-1c mRNA expression (P = 0.0162) proportionally to the degree of reduction of tumor necrosis factor alpha (TNF-alpha) mRNA (P = 0.012). Recombinant TNF-alpha almost completely abolished adipose tissue SREBP-1c mRNA expression in vivo. The t10,c12-CLA diet promoted insulin resistance and increased serum glucose (P = 0.025) and insulin (P = 0.01) concentrations. T10, c12-CLA induced profound weight loss (P = 0.0001) and increased brown and white adipose tissue UCP-2 (P = 0.001) and skeletal muscle UCP-3 (P = 0.008) mRNA expression. This study highlights the contrasting molecular and metabolic effect of two isomers of the same fatty acids. The ameliorative effect of c9,t11-CLA on lipid metabolism may be ascribed to reduced synthesis and cleavage of hepatic SREBP-1, which in turn may be regulated by hepatic LXRalpha expression. PMID- 12086932 TI - Role of peroxisome proliferator-activated receptor-gamma in maintenance of the characteristics of mature 3T3-L1 adipocytes. AB - Peroxisome proliferator-activated receptor (PPAR)-gamma plays an important role in adipogenesis. However, the functions of PPAR-gamma in differentiated adipocytes have remained unclear. The role of PPAR-gamma in mature 3T3-L1 adipocytes was therefore investigated by overexpression of a dominant negative mutant of this protein (PPAR-gamma-DeltaC) that lacks the 16 COOH-terminal amino acids and that has been shown to prevent the thiazolidinedione-induced differentiation of 3T3-L1 cells into adipocytes. Overexpression of PPAR-gamma DeltaC in mature 3T3-L1 adipocytes by adenovirus gene transfer resulted in a decrease in both cell size and intracellular triglyceride content, an increase in the extent of lipolysis, and a reduction in the rate of free fatty acid uptake. Furthermore, overexpression of this mutant reduced the abundance of mRNAs for several key enzymes that contribute to triglyceride and free fatty acid metabolism as well as the amounts of GLUT4, insulin receptor, insulin receptor substrate (IRS), and C/EBPalpha mRNAs. It also reduced both the concentration of IRS2 and the insulin-stimulated glucose uptake. These results suggest that PPAR gamma plays an important role in mature 3T3-L1 adipocytes at least in part by maintaining the expression of genes that confer the characteristics of mature adipocytes. PMID- 12086933 TI - Glucokinase is the likely mediator of glucosensing in both glucose-excited and glucose-inhibited central neurons. AB - Specialized neurons utilize glucose as a signaling molecule to alter their firing rate. Glucose-excited (GE) neurons increase and glucose-inhibited (GI) neurons reduce activity as ambient glucose levels rise. Glucose-induced changes in the ATP-to-ADP ratio in GE neurons modulate the activity of the ATP-sensitive K(+) channel, which determines the rate of cell firing. The GI glucosensing mechanism is unknown. We postulated that glucokinase (GK), a high-Michaelis constant (K(m)) hexokinase expressed in brain areas containing populations of GE and GI neurons, is the controlling step in glucosensing. Double-label in situ hybridization demonstrated neuron-specific GK mRNA expression in locus ceruleus norepinephrine and in hypothalamic neuropeptide Y, pro-opiomelanocortin, and gamma-aminobutyric acid neurons, but it did not demonstrate this expression in orexin neurons. GK mRNA was also found in the area postrema/nucleus tractus solitarius region by RT PCR. Intracarotid glucose infusions stimulated c-fos expression in the same areas that expressed GK. At 2.5 mmol/l glucose, fura-2 Ca(2+) imaging of dissociated ventromedial hypothalamic nucleus neurons demonstrated GE neurons whose intracellular Ca(2+) oscillations were inhibited and GI neurons whose Ca(2+) oscillations were stimulated by four selective GK inhibitors. Finally, GK expression was increased in rats with impaired central glucosensing (posthypoglycemia and diet-induced obesity) but was unaffected by a 48-h fast. These data suggest a critical role for GK as a regulator of glucosensing in both GE and GI neurons in the brain. PMID- 12086934 TI - Normalization of skeletal muscle glycogen synthesis and glycolysis in rosiglitazone-treated Zucker fatty rats: an in vivo nuclear magnetic resonance study. AB - The aim of this study was to characterize insulin-stimulated skeletal muscle glucose metabolism in Zucker fatty rats and to provide insight into the therapeutic mechanism by which rosiglitazone increases insulin-stimulated glucose disposal in these rats. Metabolic parameters were measured using combined in vivo (13)C nuclear magnetic resonance (NMR) spectroscopy to measure skeletal muscle glucose uptake and its distributed fluxes (glycogen synthesis and glycolysis), and (31)P NMR was used to measure simultaneous changes in glucose-6-phosphate (G 6-P) during a euglycemic-hyperinsulinemic clamp in awake Zucker fatty rats. Three groups of Zucker fatty rats (fatty rosiglitazone [FRSG], fatty control [FC], lean control [LC]) were treated for 7 days before the experiment (3 mg/kg rosiglitazone or vehicle via oral gavage). Rates of glycolysis and glycogen synthesis were assessed after treatment by monitoring 1,6-(13)C(2) glucose label incorporation into 1-(13)C glycogen, 3-(13)C lactate, and 3-(13)C alanine during a euglycemic ( approximately 7-8 mmol/l)-hyperinsulinemic (10 mU. kg(-1). min( 1)) clamp. The FRSG group exhibited a significant increase in insulin sensitivity, reflected by an increased whole-body glucose disposal rate during the clamp (24.4 +/- 1.9 vs. 17.6 +/- 1.4 and 33.2 +/- 2.0 mg. kg(-1). min(-1) in FRSG vs. FC [P < 0.05] and LC [P < 0.01] groups, respectively). The increased insulin-stimulated glucose disposal in the FRSG group was associated with a normalization of the glycolytic flux (52.9 +/- 9.1) to LC (56.2 +/- 16.6) versus FC (18.8 +/- 8.6 nmol. g(-1). min(-1), P < 0.02) and glycogen synthesis flux (56.3 +/- 11.5) to LC (75.2 +/- 15.3) versus FC (16.6 +/- 12.8 nmol. g(-1). min( 1), P < 0.05). [G-6-P] increased in the FRSG and LC groups versus baseline during the clamp (13.0 +/- 11.1 and 16.9 +/- 5.8%, respectively), whereas [G-6-P] in the FC group decreased (-23.3 +/- 13.4%, P < 0.05). There were no differences between groups in intramyocellular glucose, as measured by biochemical assay. These data suggest that the increased insulin-stimulated glucose disposal in muscle after rosiglitazone treatment can be attributed to a normalization of glucose transport and metabolism. PMID- 12086936 TI - Improved insulin sensitivity is associated with restricted intake of dietary glycoxidation products in the db/db mouse. AB - Advanced glycation end products (AGEs), known promoters of diabetic complications, form abundantly in heated foods and are ingested in bioreactive forms. To test whether dietary AGEs play a role in the progression of insulin resistance, C57/BL/KsJ db/db mice were randomly placed for 20 weeks on a diet with either a low AGE content (LAD) or a 3.4-fold higher content of AGE (high AGE diet [HAD]), including (epsilon)N-carboxymethyllysine (CML) and methylglyoxal (MG). LAD-fed mice showed lower fasting plasma insulin levels throughout the study (P = 0.01). Body weight was reduced by approximately 13% compared with HAD fed mice (P = 0.04) despite equal food intake. LAD-fed mice exhibited significantly improved responses to both glucose (at 40 min, P = 0.003) and insulin (at 60 min, P = 0.007) tolerance tests, which correlated with a twofold higher glucose uptake by adipose tissue (P = 0.02). Compared with the severe hypertrophy and morphological disorganization of islets from HAD-fed mice, LAD fed mice presented a better-preserved structure of the islets. LAD-fed mice demonstrated significantly increased plasma HDL concentrations (P < 0.0001). Consistent with these observations, LAD-fed mice exhibited twofold lower serum CML and MG concentrations compared with HAD-fed mice (P = 0.02). These results demonstrate that reduced AGE intake leads to lower levels of circulating AGE and to improved insulin sensitivity in db/db mice. PMID- 12086935 TI - Metformin increases AMP-activated protein kinase activity in skeletal muscle of subjects with type 2 diabetes. AB - Metformin is an effective hypoglycemic drug that lowers blood glucose concentrations by decreasing hepatic glucose production and increasing glucose disposal in skeletal muscle; however, the molecular site of metformin action is not well understood. AMP-activated protein kinase (AMPK) activity increases in response to depletion of cellular energy stores, and this enzyme has been implicated in the stimulation of glucose uptake into skeletal muscle and the inhibition of liver gluconeogenesis. We recently reported that AMPK is activated by metformin in cultured rat hepatocytes, mediating the inhibitory effects of the drug on hepatic glucose production. In the present study, we evaluated whether therapeutic doses of metformin increase AMPK activity in vivo in subjects with type 2 diabetes. Metformin treatment for 10 weeks significantly increased AMPK alpha2 activity in the skeletal muscle, and this was associated with increased phosphorylation of AMPK on Thr172 and decreased acetyl-CoA carboxylase-2 activity. The increase in AMPK alpha2 activity was likely due to a change in muscle energy status because ATP and phosphocreatine concentrations were lower after metformin treatment. Metformin-induced increases in AMPK activity were associated with higher rates of glucose disposal and muscle glycogen concentrations. These findings suggest that the metabolic effects of metformin in subjects with type 2 diabetes may be mediated by the activation of AMPK alpha2. PMID- 12086938 TI - Appetite suppression and weight reduction by a centrally active aminosterol. AB - The rise in obesity and its complications has generated enormous interest in the regulation of feeding and body weight. We show that a spermine metabolite of cholesterol (MSI-1436) decreases body weight, specifically fat, by suppressing feeding and preventing the reduction in energy expenditure, hormonal changes, and patterns of neuropeptide expression normally associated with weight loss. MSI 1436 enters the brain after peripheral injection and is more potent when injected into the cerebral ventricle (intracerebroventricular [ICV]). Systemic or ICV MSI 1436 administration induced similar patterns of Fos immunoreactivity in the brain, especially the paraventricular hypothalamic nucleus (PVN). This brain region integrates neural signals from hypothalamic and brain stem nuclei and regulates feeding behavior, autonomic function, and neuroendocrine function. Microinjection of MSI-1436 into the PVN potently suppressed feeding and reduced body weight for several days. Unlike caloric restriction, MSI-1436 decreased mRNA levels of agouti-related peptide and neuropeptide Y in the hypothalamus. These findings indicate that MSI-1436 acts in the brain to regulate food intake and energy expenditure, likely through suppression of orexigenic hypothalamic pathways. PMID- 12086937 TI - Sustained exposure of L6 myotubes to high glucose and insulin decreases insulin stimulated GLUT4 translocation but upregulates GLUT4 activity. AB - Hyperglycemia and hyperinsulinemia are cardinal features of acquired insulin resistance. In adipose cell cultures, high glucose and insulin cause insulin resistance of glucose uptake, but because of altered GLUT4 expression and contribution of GLUT1 to glucose uptake, the basis of insulin resistance could not be ascertained. Here we show that GLUT4 determines glucose uptake in L6 myotubes stably overexpressing myc-tagged GLUT4. Preincubation for 24 h with high glucose and insulin (high Glc/Ins) reduced insulin-stimulated GLUT4 translocation by 50%, without affecting GLUT4 expression. Insulin receptor and insulin receptor substrate-1 tyrosine phosphorylation, phosphatidylinositol 3-kinase activation, and Akt phosphorylation also diminished, as did insulin-mediated glucose uptake. However, basal glucose uptake rose by 40% without any gain in surface GLUT4. High Glc/Ins elevated basal p38 mitogen-activated protein kinase (MAPK) phosphorylation and activity, and a short inhibition of p38 MAPK with SB202190 corrected the rise in basal glucose uptake, suggesting that p38 MAPK activity contributes to this rise. We propose that in a cellular model of skeletal muscle, chronic exposure to high Glc/Ins reduced the acute, insulin-elicited GLUT4 translocation. In addition, basal state GLUT4 activity was augmented to partially compensate for the translocation defect, resulting in a more robust glucose uptake than what would be predicted from the amount of cell surface GLUT4 alone. PMID- 12086939 TI - Regulation of circulating soluble leptin receptor levels by gender, adiposity, sex steroids, and leptin: observational and interventional studies in humans. AB - Leptin is an adipocyte-secreted hormone important in energy homeostasis and diverse physiological processes. A circulating soluble form of the leptin receptor [soluble leptin receptor (sOB-R)] is the main leptin-binding protein and determinant of free leptin index (FLI), the presumed biologically active form of leptin. We performed observational and interventional studies to elucidate the regulation of sOB-R and FLI in humans. In a cross-sectional study (n = 118), leptin, gender, and adiposity were significant determinants of sOB-R. By multivariate analysis, estradiol (E2) and testosterone predict sOB-R, whereas insulin predicts leptin and FLI. In a frequent-sampling study (n = 6), sOB-R followed a significant circadian rhythm inverse to that of leptin, suggesting that leptin's biological activity may have an even more pronounced diurnal variation than originally thought. A 72-h fast in eight men decreased leptin levels by 80% and increased lymphocyte expression of leptin receptor mRNA and serum sOB-R levels by 100%. Physiological and pharmacological doses of recombinant-methionyl human leptin (rhLeptin) administered to fasted men prevented the fasting-induced increase of sOB-R levels, and pharmacological doses resulted in a decrease in sOB-R levels. These studies provide evidence that sOB-R is regulated by gender, adiposity, hormones, and rhLeptin administration. This may have important implications for the biological activity of leptin in disease states associated with abnormal leptin levels (e.g., obesity and anorexia nervosa). PMID- 12086940 TI - Adrenalectomy improves diabetes in A-ZIP/F-1 lipoatrophic mice by increasing both liver and muscle insulin sensitivity. AB - The virtually fatless A-ZIP/F-1 mouse is profoundly insulin resistant, diabetic, and a good model for humans with severe generalized lipoatrophy. Like a number of other mouse models of diabetes, the A-ZIP/F-1 mouse has elevated serum corticosterone levels. Leptin infusion lowers the corticosterone levels, suggesting that leptin deficiency contributes to the hypercorticosteronemic state. To test the hypothesis that the increased glucocorticoids contribute to the diabetes and insulin resistance, we examined the effect of adrenalectomy on A ZIP/F-1 mice. Adrenalectomy significantly decreased the blood glucose, serum insulin, and glycated hemoglobin levels. Hyperinsulinemic-euglycemic clamps were performed to characterize the changes in whole-body and tissue insulin sensitivity. The adrenalectomized A-ZIP/F-1 mice displayed a marked improvement in insulin-induced suppression of endogenous glucose production, indicating increased hepatic insulin sensitivity. Adrenalectomy also increased muscle glucose uptake and glycogen synthesis. These results suggest that the chronically increased serum corticosterone levels contribute to the diabetes of the A-ZIP/F-1 mice and that removal of the glucocorticoid excess improves the insulin sensitivity in both muscle and liver. PMID- 12086941 TI - Redirection of human autoreactive T-cells Upon interaction with dendritic cells modulated by TX527, an analog of 1,25 dihydroxyvitamin D(3). AB - The active form of vitamin D(3), 1alpha,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), is a potent immunomodulator known to affect T-cells through targeting antigen-presenting cells such as dendritic cells (DCs). We studied the effects of a novel nonhypercalcemic 1,25(OH)(2)D(3) analog, TX527, on DC differentiation, maturation, and function with respect to stimulation of a committed human GAD65-specific autoreactive T-cell clone. Continuous addition of TX527 impaired interleukin (IL)-4 and granulocyte/macrophage colony-stimulating factor (GM-CSF)-driven DC differentiation as well as lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma)-induced maturation into Th1-promoting DC (DC1), as characterized by marked changes in DC morphology and abrogation of IL-12p70 release upon CD40 ligation. Addition of TX527 during maturation did not affect DC morphology but significantly changed DC cytokine profiles. The potential of treated DCs to alter the response pattern of committed autoreactive T-cells was found to depend on the timing of TX527 exposure. Continuously TX527-treated DCs significantly inhibited T-cell proliferation and blocked IFN-gamma, IL-10, but not IL-13 production, whereas DCs treated during maturation failed to inhibit T cell proliferation but affected IL-10 and IFN-gamma production. Collectively, we provide evidence that nonhypercalcemic TX527 is a potent in vitro DC modulator, yielding DCs with the potential to change cytokine responses of committed autoreactive T-cells. PMID- 12086942 TI - Immunological characterization and therapeutic activity of an altered-peptide ligand, NBI-6024, based on the immunodominant type 1 diabetes autoantigen insulin B-chain (9-23) peptide. AB - The nonobese diabetic (NOD) mouse is a good model for human type 1 diabetes, which is characterized by autoreactive T-cell-mediated destruction of insulin producing islet beta-cells of the pancreas. The 9-23 amino acid region of the insulin B-chain [B((9-23))] is an immunodominant T-cell target antigen in the NOD mouse that plays a critical role in the disease process. By testing a series of B((9-23)) peptide analogs with single or double alanine substitutions, we identified a set of altered peptide ligands (APLs) capable of inhibiting B((9 23))-induced proliferative responses of NOD pathogenic T-cell clones. These APLs were unable to induce proliferation of these clones. However, vaccinations with the APLs induced strong cellular responses, as measured by in vitro lymphocyte proliferation and Th2 cytokine production (i.e., interleukin [IL]-4 and IL-10, but not gamma-interferon [IFN-gamma]). These responses were cross-reactive with the native antigen, B((9-23)), suggesting that the APL-induced Th2 responses may provide protection by controlling endogenous B((9-23))-specific Th1 (i.e., IFN gamma-producing) pathogenic responses. One of these APLs that contained alanine substitutions at residues 16 and 19 (16Y-->A, 19C-->A; NBI-6024) was further characterized for its therapeutic activity because it consistently induced T-cell responses (e.g., T-cell lines and clones) that were of the Th2 type and that were cross-reactive with B((9-23)). Subcutaneous injections of NBI-6024 to NOD mice administered either before or after the onset of disease substantially delayed the onset and reduced the incidence of diabetes. This study is the first to report therapeutic activity of an APL derived from an islet beta-cell-specific antigen in type 1 diabetes. PMID- 12086943 TI - Pancreatic islet transplantation using the nonhuman primate (rhesus) model predicts that the portal vein is superior to the celiac artery as the islet infusion site. AB - We've established a nonhuman primate islet allotransplant model to address questions such as whether transplanting islets into the gut's arterial system would more safely and as effectively support long-term islet allograft survival compared with the traditional portal vein approach. We reasoned that islets make up <2% of pancreatic cell mass but consume an estimated 20% of arterial blood flow, suggesting an advantage for the arterial site. Access to the arterial system is also easier and safer than the portal system. Pancreatectomized rhesus macaques were transplanted with allogeneic islets infused into either the portal vein (n = 6) or the celiac artery (n = 4). To prevent rejection, primates were given daclizumab, tacrolimus, and rapamycin. In five of six portal vein experiments, animals achieved normoglycemia without exogenous insulin. In contrast, none of the animals given intra-arterial islets showed even transient insulin independence (P = 0.048). Two of the latter animals received a second islet transplant, this time to the portal system, and both achieved insulin independence. Thus, intraportal islet transplantation under conventional immunosuppression is feasible in primates and can result in long-term insulin independence when adequate immunosuppression is maintained. Arterial islet injection, however, does not appear to be a viable islet transplantation technique. PMID- 12086944 TI - An interval tightly linked to but distinct from the H2 complex controls both overt diabetes (Idd16) and chronic experimental autoimmune thyroiditis (Ceat1) in nonobese diabetic mice. AB - The major histocompatibility complex (MHC) has long been associated with predisposition to several autoimmune diseases, including type 1 diabetes and autoimmune thyroiditis. In type 1 diabetes, a primary role has been assigned to class II genes, both in humans and in the nonobese diabetic (NOD) mouse model. However, an involvement of other tightly linked genes is strongly suspected. Here, through two independent sets of experiments, we provide solid evidence for the existence of at least one such gene. First, using a new recombinant congenic NOD strain, R114, we definitively individualized the Idd16 locus from the MHC in a 6-cM interval proximal to H2-K. It affords almost complete protection against diabetes and is associated with delayed insulitis. Second, by genome scan, we mapped non-H2 genes associated with the highly penetrant form of chronic experimental autoimmune thyroiditis (EAT) that is elicited in NOD and NOD.H2(k) mice by immunization with thyroglobulin. We identified one major dominant locus, Ceat1, on chromosome 17, overlapping with Idd16. Most importantly, R114 recombinant congenic mice challenged with thyroglobulin did not develop chronic EAT. This new major region defined by both Idd16 and Ceat1 might thus concur to the unique strength of the MHC in autoimmune susceptibility of NOD mice. PMID- 12086945 TI - Successful islet transplantation: continued insulin reserve provides long-term glycemic control. AB - Clinical islet transplantation is gaining acceptance as a potential therapy, particularly for subjects who have labile diabetes or problems with hypoglycemic awareness. The risks of the procedure and long-term outcomes are still not fully known. We have performed 54 islet transplantation procedures on 30 subjects and have detailed follow-up in 17 consecutive Edmonton protocol-treated subjects who attained insulin independence after transplantation of adequate numbers of islets. Subjects were assessed pretransplant and followed prospectively posttransplant for immediate and long-term complications related to the procedure or immunosuppressive therapy. The 17 patients all became insulin independent after a minimum of 9,000 islets/kg were transplanted. Of 15 consecutive patients with at least 1 year of follow-up after the initial transplant, 12 (80%) were insulin independent at 1 year. In 14 subjects who have maintained demonstrable C peptide secretion, glucose control has been stable and glycemic lability and problems with hypoglycemic reactions have been corrected. After 2 of the 54 procedures, some thrombosis was detected in the portal vein circulation. Five subjects had bleeding related to the percutaneous portal vein access procedures: three required transfusion alone, and in one subject, who had a partial thrombosis of the portal vein, an expanding intrahepatic and subscapular hemorrhage occurred while on anticoagulation, requiring transfusion and surgery. Elevated liver function test results were found in 46% of subjects but resolved in all. Complications related to the therapy have been hypercholesterolemia requiring statin therapy in 65%; a rise in creatinine in two patients, both of whom had preexisting renal disease; a rise in protein in four, all of whom had preexisting proteinuria; and antihypertensive therapy increased or started in 53%. Three of the 17 patients have required retinal laser photocoagulation. There have been no cases of posttransplant lymphoproliferative disorder or cytomegalovirus infection, and no deaths. The acute insulin response to arginine correlated better with transplanted islet mass than acute insulin response to glucose (AIR(g)) and area under the curve for insulin (AUC(i)), but the AIR(g) and AUC(i) were more closely related to glycemic control. The AUC(i) directly posttransplant was lower in those who eventually became C-peptide deficient. Our results, with a maximum follow-up of 34 months, indicate that prolonged insulin independence can be achieved after islet transplantation. There are some risks associated acutely with the procedure, and hypercholesterolemia and hypertension are treatable concerns on longer-term follow-up. All patients with persisting C peptide secretion have had a resolution of both glycemic lability and problems with hypoglycemic reactions. Apart from the rise in serum creatinine in two subjects, no serious consequences of immunosuppressive therapy have been encountered. Islet transplantation is a reasonable option in those with severe problems with glycemic lability or hypoglycemia. PMID- 12086946 TI - Estrogen can prevent or reverse obesity and diabetes in mice expressing human islet amyloid polypeptide. AB - Type 2 diabetes is characterized by loss of beta-cell mass and concomitant deposition of amyloid derived from islet amyloid polypeptide (IAPP). Previously we have shown that expression of human IAPP (huIAPP) in islets of transgenic mice results in either a rapid onset of hyperglycemia in mice homozygous for the huIAPP transgene on a lean background (FVB/N) or a gradual hyperglycemia in mice hemizygous for the huIAPP transgene on an obese background (A(vy)/A). In both strains, only the males routinely develop diabetes. To investigate this sexual dimorphism, we treated young prediabetic A(vy)/A mice transgenic for huIAPP (huIAPP-A(vy)) with 17beta-estradiol (E2). The treatment completely blocked the progression to hyperglycemia but also prevented the associated weight gain in these mice. Immunohistochemistry of pancreatic sections demonstrated normal islet morphology with no apparent deposition of islet amyloid. E2 treatment of 1-year old huIAPP-A(vy) diabetic males rapidly reverses obesity and hyperglycemia. To determine the effects of E2 in a nonobese model, we also treated prediabetic, ad libitum-fed and pair-fed Lean-huIAPP transgenic males. E2 completely blocked the progression to hyperglycemia with no significant effect on body weight. Pancreatic insulin content and plasma insulin concentration of Lean-huIAPP transgenic mice increased in a dose-dependent manner. We demonstrated the presence of estrogen receptor (ER)-alpha mRNA in mouse and human islets. By also confirming the presence of ER-alpha protein in islets, we discovered a novel 58 kDa ER-alpha isoform in mice and a 52-kDa isoform in humans, in the absence of the classic 67-kDa protein found in most tissues of both species. The demonstrated presence of ER-alpha in mouse and human islets is consistent with a direct effect on islet function. We conclude that exogenous E2 administered to male mice may block human IAPP-mediated beta-cell loss both by direct action on beta-cells and by decreasing insulin demand through inhibition of weight gain or increasing insulin action. PMID- 12086947 TI - Beta-cell function is a major contributor to oral glucose tolerance in high-risk relatives of four ethnic groups in the U.S. AB - First-degree relatives of individuals with type 2 diabetes are at increased risk of developing hyperglycemia. To examine the prevalence and pathogenesis of abnormal glucose homeostasis in these subjects, 531 first-degree relatives with no known history of diabetes (aged 44.1 +/- 0.7 years; BMI 29.0 +/- 0.3 kg/m(2)) underwent an oral glucose tolerance test (OGTT). Newly identified diabetes was found in 19% (n = 100), and impaired fasting glucose (IFG) and/or impaired glucose tolerance (IGT) was found in 36% (n = 191). Thus, only 45% (n = 240) had normal glucose tolerance (NGT). The homeostasis model assessment of insulin resistance (HOMA-IR) was used to estimate insulin sensitivity; beta-cell function was quantified as the ratio of the incremental insulin to glucose responses over the first 30 min during the OGTT (DeltaI(30)/DeltaG(30)). This latter measure was also adjusted for insulin sensitivity as it modulates beta-cell function ([DeltaI(30)/DeltaG(30)]/HOMA-IR). Decreasing glucose tolerance was associated with increasing insulin resistance (HOMA: NGT 12.01 +/- 0.54 pmol/mmol; IFG/IGT 16.14 +/- 0.84; diabetes 26.99 +/- 2.62; P < 0.001) and decreasing beta-cell function (DeltaI(30)/DeltaG(30): NGT 157.7 +/- 9.7 pmol/mmol; IFG/IGT 100.4 +/- 5.4; diabetes 57.5 +/- 7.3; P < 0.001). Decreasing beta-cell function was also identified when adjusting this measure for insulin sensitivity ([DeltaI(30)/DeltaG(30)]/HOMA-IR). In all four ethnic groups (African-American, n = 55; Asian-American, n = 66; Caucasian, n = 217; Hispanic-American, n = 193), IFG/IGT and diabetic subjects exhibited progressively increasing insulin resistance and decreasing beta-cell function. The relationships of insulin sensitivity and beta-cell function to glucose disposal, as measured by the incremental glucose area under the curve (AUCg), were examined in the whole cohort. Insulin sensitivity and AUCg were linearly related so that insulin resistance was associated with poorer glucose disposal (r(2) = 0.084, P < 0.001). In contrast, there was a strong inverse curvilinear relationship between beta cell function and AUCg such that poorer insulin release was associated with poorer glucose disposal (log[DeltaI(30)/DeltaG(30)]: r(2) = 0.29, P < 0.001; log[(DeltaI(30)/DeltaG(30))/HOMA-IR]: r(2) = 0.45, P < 0.001). Thus, abnormal glucose metabolism is common in first-degree relatives of subjects with type 2 diabetes. Both insulin resistance and impaired beta-cell function are associated with impaired glucose metabolism in all ethnic groups, with beta-cell function seeming to be more important in determining glucose disposal. PMID- 12086948 TI - Glycemic control determines hepatic and peripheral glucose effectiveness in type 2 diabetic subjects. AB - Glucose effectiveness is impaired in type 2 diabetes. We hypothesize that chronic hyperglycemia and hyperlipidemia contribute importantly to this defect. To test this hypothesis, we compared the effect of acute hyperglycemia on glucose turnover in type 2 diabetic subjects in good control (GC) (n = 14, age 51.7 +/- 3.7 years, BMI 28.4 +/- 1.0 kg/ m(2), HbA(1c) 5.9 +/- 0.2%) and poor control (PC) (n = 10, age 50.0 +/- 2.5 years, BMI 27.9 +/- 1.5 kg/m(2), HbA(1c) 9.9 +/- 0.6%) with age- and weight-matched nondiabetic subjects (ND) (n = 11, age 47.0 +/- 4.4 years, BMI 28.5 +/- 1.0 kg/m(2), HbA(1c) 5.1 +/- 0.2%). Fixed hormonal conditions were attained by infusing somatostatin for 6 h with replacement of basal insulin, glucagon, and growth hormone. Glucose fluxes ([3-(3)H]glucose) were compared during euglycemic (5 mmol/l, t = 180-240 min) and hyperglycemic (Hy) (10 mmol/l, t = 300-360 min, variable glucose infusion) clamp intervals. Acute hyperglycemia suppressed hepatic glucose production (GP) by 43% and increased peripheral glucose uptake (GU) by 86% in the ND subjects. Conversely, GP failed to suppress (-7%) and GU was suboptimally increased (+34%) in response to Hy in the PC group. However, optimal glycemic control was associated with normal glucose effectiveness in GC subjects (GP -38%, GU +72%; P > 0.05 for GC vs. ND). To determine whether short-term correction of hyperglycemia and/or hyperlipidemia is sufficient to reverse the impairment in glucose effectiveness, five PC subjects were restudied after 72 h of normoglycemia ( approximately 100 mg/dl; variable insulin infusions). These subjects regained normal effectiveness of glucose to suppress GP and stimulate GU and in response to Hy (GP -47%, GU + 71%; P > 0.05 vs. baseline studies). Thus, chronic hyperglycemia and/or hyperlipidemia contribute to impaired effectiveness of glucose in regulating glucose fluxes in type 2 diabetes and hence to worsening of the overall metabolic condition. Short term normalization of plasma glucose might break the vicious cycle of impaired glucose effectiveness in type 2 diabetes. PMID- 12086949 TI - Inhibition of glycogen synthase kinase 3 improves insulin action and glucose metabolism in human skeletal muscle. AB - Glycogen synthase kinase (GSK)-3 has been implicated in the regulation of multiple cellular physiological processes in skeletal muscle. Selective cell permeable reversible inhibitors (INHs) of GSK-3 (CT98014 and CHIR98023 [Chiron, Emeryville, CA] and LiCl) were used to evaluate the role of GSK-3 in controlling glucose metabolism. Acute treatment (30 min) of cultured human skeletal muscle cells with either INH resulted in a dose-dependent activation of glycogen synthase (GS) with a maximally effective concentration of approximately 2 micromol/l. The maximal acute effect of either INH on GS (103 +/- 25% stimulation over basal) was greater than the maximal insulin response (48 +/- 9%, P < 0.05 vs. INH); LiCl was as effective as insulin. The GSK-3 inhibitor effect, like that of insulin, was on the activation state (fractional velocity [FV]) of GS. Cotreatment of muscle cells with submaximal doses of INH and insulin resulted in an additive effect on GS FV (103 +/- 10% stimulation, P < 0.05 vs. either agent alone). Glucose incorporation into glycogen was also acutely stimulated by INH. While prolonged (6-24 h) insulin exposure led to desensitization of GS, INH continued to activate GS FV for at least 24 h. Insulin and LiCl acutely activated glucose uptake, whereas INH stimulation of glucose uptake required more prolonged exposure, starting at 6 h and continuing to 24 h. Chronic (4-day) treatment with INH increased both basal (154 +/- 32% of control) and insulin-stimulated (219 +/- 74%) glucose uptake. Upregulation of uptake activity occurred without any change in total cellular GLUT1 or GLUT4 protein content. Yet the same chronic treatment resulted in a 65 +/- 6% decrease in GSK-3 protein and a parallel decrease (61 +/- 11%) in GSK-3 total activity. Together with the INH-induced increase in insulin stimulated glucose uptake, there was an approximately 3.5-fold increase (P < 0.05) in insulin receptor substrate (IRS)-1 protein abundance. Despite upregulation of IRS-1, maximal insulin stimulation of Akt phosphorylation was unaltered by INH treatment. The results suggest that selective inhibition of GSK 3 has an impact on both GS and glucose uptake, including effects on insulin action, using mechanisms that differ from and are additive to those of insulin. PMID- 12086950 TI - Long-term AICAR administration reduces metabolic disturbances and lowers blood pressure in rats displaying features of the insulin resistance syndrome. AB - The insulin resistance syndrome is characterized by several risk factors for cardiovascular disease. Chronic chemical activation of AMP-activated protein kinase by the adenosine analog 5-aminoimidazole-4-carboxamide-1-beta -D ribofuranoside (AICAR) has been shown to augment insulin action, upregulate mitochondrial enzymes in skeletal muscles, and decrease the content of intra abdominal fat. Furthermore, acute AICAR exposure has been found to reduce sterol and fatty acid synthesis in rat hepatocytes incubated in vitro as well as suppress endogenous glucose production in rats under euglycemic clamp conditions. To investigate whether chronic AICAR administration, in addition to the beneficial effects on insulin sensitivity, is capable of improving other phenotypes associated with the insulin resistance syndrome, obese Zucker (fa/fa) rats (n = 6) exhibiting insulin resistance, hyperlipidemia, and hypertension were subcutaneously injected with AICAR (0.5 mg/g body wt) daily for 7 weeks. Obese control rats were either pair-fed (PF) (n = 6) or ad libitum-fed (AL) (n = 6). Lean Zucker rats (fa/-) (n = 8) served as a reference group. AICAR administration significantly reduced plasma triglyceride levels (P < 0.01 for AICAR vs. AL, and P = 0.05 for AICAR vs. PF) and free fatty acids (P < 0.01 for AICAR vs. AL, and P < 0.05 for AICAR vs. PF) and increased HDL cholesterol levels (P < 0.01 for AICAR vs. AL and PF). AICAR treatment also lowered systolic blood pressure by 14.6 +/- 4.3 mmHg (P < 0.05), and AICAR-treated animals exhibited a tendency toward decreased intra-abdominal fat content. Furthermore, AICAR administration normalized the oral glucose tolerance test and decreased fasting concentrations of glucose and insulin close to the level of the lean animals. Finally, in line with previous findings, AICAR treatment was also found to enhance GLUT4 protein expression and to increase maximally insulin-stimulated glucose transport in primarily white fast-twitch muscles. Our data provide strong evidence that long term administration of AICAR improves glucose tolerance, improves the lipid profile, and reduces systolic blood pressure in an insulin-resistant animal model. The present study gives additional support to the hypothesis that AMPK activation might be a potential future pharmacological strategy for treating the insulin resistance syndrome. PMID- 12086951 TI - TNF-alpha is a predictor of insulin resistance in human pregnancy. AB - Historically, insulin resistance during pregnancy has been ascribed to increased production of placental hormones and cortisol. The purpose of this study was to test this hypothesis by correlating the longitudinal changes in insulin sensitivity during pregnancy with changes in placental hormones, cortisol, leptin, and tumor necrosis factor (TNF)-alpha. Insulin resistance was assessed in 15 women (5 with gestational diabetes mellitus [GDM] and 10 with normal glucose tolerance) using the euglycemic-hyperinsulinemic clamp procedure, before pregnancy (pregravid) and during early (12-14 weeks) and late (34-36 weeks) gestation. Body composition, plasma TNF-alpha, leptin, cortisol, and reproductive hormones (human chorionic gonadotropin, estradiol, progesterone, human placental lactogen, and prolactin) were measured in conjunction with the clamps. Placental TNF-alpha was measured in vitro using dually perfused human placental cotyledon from five additional subjects. Compared with pregravid, insulin resistance was evident during late pregnancy in all women (12.4 +/- 1.2 vs. 8.1 +/- 0.8 10(-2) mg. kg(-1) fat-free mass. min(-1). microU(-1). ml(-1)). TNF-alpha, leptin, cortisol, all reproductive hormones, and fat mass were increased in late pregnancy (P < 0.001). In vitro, most of the placental TNF-alpha (94%) was released into the maternal circulation; 6% was released to the fetal side. During late pregnancy, TNF-alpha was inversely correlated with insulin sensitivity (r = 0.69, P < 0.006). Furthermore, among all of the hormonal changes measured in this study, the change in TNF-alpha from pregravid to late pregnancy was the only significant predictor of the change in insulin sensitivity (r = -0.60, P < 0.02). The placental reproductive hormones and cortisol did not correlate with insulin sensitivity in late pregnancy. Multivariate stepwise regression analysis revealed that TNF-alpha was the most significant independent predictor of insulin sensitivity (r = -0.67, P < 0.0001), even after adjustment for fat mass by covariance (r = 0.46, P < 0.01). These observations challenge the view that the classical reproductive hormones are the primary mediators of change in insulin sensitivity during gestation and provide the basis for including TNF-alpha in a new paradigm to explain insulin resistance in pregnancy. PMID- 12086952 TI - Alterations in the activity of placental amino acid transporters in pregnancies complicated by diabetes. AB - Alterations in placental transport may contribute to accelerated fetal growth in pregnancies complicated by diabetes. We studied the activity of the syncytiotrophoblast amino acid transporter system A and the transport of the essential amino acids leucine, lysine, and taurine. Syncytiotrophoblast microvillous plasma membranes (MVMs) and basal plasma membranes (BMs) were isolated from placentas obtained from normal pregnancies and pregnancies complicated by gestational diabetes mellitus (GDM) and type 1 diabetes, with and without large-for-gestational-age (LGA) fetuses. Amino acid transport was assessed using radio-labeled substrates and rapid filtration techniques. System A activity in MVM was increased (65-80%, P < 0.05) in all groups with diabetes independent of fetal overgrowth. However, MVM system A activity was unaffected in placentas of normal pregnancies with LGA fetuses. MVM leucine transport was increased in the GDM/LGA group. In BMs, amino acid transport was unaffected by diabetes. In conclusion, diabetes in pregnancy is associated with an increased system A activity in MVM, and MVM leucine transport is increased in the GDM/LGA group. We suggest that these changes result in an increased uptake of neutral amino acids across MVM, which may be used in placental metabolism or be delivered to the fetus. The increased MVM leucine uptake in the GDM/LGA group may contribute to accelerated fetal growth in these patients. PMID- 12086953 TI - The effect of a 3-month low-intensity endurance training program on fat oxidation and acetyl-CoA carboxylase-2 expression. AB - Endurance training has been shown to increase fat oxidation both at rest and during exercise. However, most exercise training studies have been performed at high exercise intensity in well-trained athletes, and not much is known about the effect of a low-intensity training program on fat oxidation capacity in lean sedentary humans. Here, we examine the effect of 3-month low-intensity training program on total and intramuscular triglyceride (IMTG)- and/or VLDL-derived fat oxidation capacity and skeletal muscle mRNA expression. Six healthy untrained subjects (aged 43 +/- 2 years, BMI 22.7 +/- 1.1 kg/ m(2), V(O)(2max) 3.2 +/- 0.2 l/min) participated in a supervised 12-week training program at 40% V(O)(2max) three times weekly. Total and plasma-derived fatty acid oxidation at rest and during 1 h exercise was measured using [(13)C]palmitate, and in a separate test, [(13)C]acetate recovery was determined. Muscle biopsies were taken after an overnight fast. Total fat oxidation during exercise increased from 1,241 +/- 93 to 1,591 +/- 130 micromol/min (P = 0.06), and IMTG- and/or VLDL-derived fatty acid oxidation increased from 236 +/- 84 to 639 +/- 172 micromol/min (P = 0.09). Acetyl-CoA carboxylase-2 mRNA expression was significantly decreased after training (P = 0.005), whereas lipoprotein lipase mRNA expression tended to increase (P = 0.07). In conclusion, a minimal amount of physical activity tends to increase fat oxidation and leads to marked changes in the expression of genes encoding for key enzymes in fat metabolism. PMID- 12086954 TI - Herpes simplex-mediated gene transfer of nerve growth factor protects against peripheral neuropathy in streptozotocin-induced diabetes in the mouse. AB - Peripheral neuropathy is a common and debilitating complication of diabetes. In animal models, neurotrophic factors can prevent progression of the neuropathy, but adverse effects prevent systemic administration in adequate doses to treat human disease. We examined whether gene transfer with replication-defective genomic herpes simplex virus (HSV) vectors modified to express nerve growth factor (NGF) could be used to prevent progression of neuropathy in mice. Diabetes induced by streptozotocin (STZ) resulted in a sensory neuropathy manifest by a decrease in the foot sensory nerve amplitude (FSA; control = 20 +/- 0.1 microV, treated = 14 +/- 0.1 microV). Transduction of dorsal root ganglia in vivo with an HSV-based vector expressing NGF under the control of the human cytomegalovirus immediate early promoter (vector SHN) or the HSV latency active promoter 2 (vector SLN) by footpad inoculation 2 weeks after STZ administration protected against the decrease in FSA (22 +/- 1.4 microV and 21 +/- 1.7 microV, respectively) measured 4 weeks later. Injection of SHN into inguinal adipose tissue 2 weeks after onset of diabetes also prevented the decrease in FSA (20 +/- 3.3 microV). These results suggest that gene transfer with an NGF-producing herpes-based vector may prove useful in the treatment of diabetic neuropathy. PMID- 12086955 TI - Influence of experimental diabetes on the microcirculation of injured peripheral nerve: functional and morphological aspects. AB - Regeneration of diabetic axons has delays in onset, rate, and maturation. It is possible that microangiopathy of vasa nervorum, the vascular supply of the peripheral nerve, may render an unfavorable local environment for nerve regeneration. We examined local nerve blood flow proximal and distal to sciatic nerve transection in rats with long-term (8 month) experimental streptozotocin diabetes using laser Doppler flowmetry and microelectrode hydrogen clearance polarography. We then correlated these findings, using in vivo perfusion of an India ink preparation, by outlining the lumens of microvessels from unfixed nerve sections. There were no differences in baseline nerve blood flow between diabetic and nondiabetic uninjured nerves, and vessel number, density, and area were unaltered. After transection, there were greater rises in blood flow in proximal stumps of nondiabetic nerves than in diabetic animals associated with a higher number, density, and caliber of epineurial vessels. Hyperemia also developed in distal stumps of nondiabetic nerves but did not develop in diabetic nerves. In these stumps, diabetic rats had reduced vessel numbers and smaller mean endoneurial vessel areas. Failed or delayed upregulation of nerve blood flow after peripheral nerve injury in diabetes may create a relatively ischemic regenerative microenvironment. PMID- 12086956 TI - Activation of nuclear factor-kappaB induced by diabetes and high glucose regulates a proapoptotic program in retinal pericytes. AB - To reconstruct the events that may contribute to the accelerated death of retinal vascular cells in diabetes, we investigated in situ and in vitro the activation of nuclear factor-kappaB (NF-kappaB), which is triggered by cellular stress and controls several programs of gene expression. The retinal capillaries of diabetic eye donors showed an increased number of pericyte nuclei positive for NF-kappaB, when compared with nondiabetic donors, whereas endothelial cells were negative. Microvascular cell apoptosis and acellular capillaries were increased only in the diabetic donors with numerous NF-kappaB-positive pericytes. Likewise, high glucose in vitro activated NF-kappaB in retinal pericytes but not in endothelial cells, and increased apoptosis only in pericytes. Studies with NF-kappaB inhibitors suggested that in pericytes, basal NF-kappaB has prosurvival functions, whereas NF-kappaB activation induced by high glucose is proapoptotic. Pericytes exposed to high glucose showed increased expression of Bax and of tumor necrosis factor-alpha, which were prevented by the NF-kappaB inhibitors and mimicked by transfection with the p65 subunit of NF-kappaB, and failed to increase the levels of the NF-kappaB-dependent inhibitors of apoptosis. Colocalization of activated NF-kappaB and Bax overexpression was observed in the retinal pericytes of diabetic donors. A proapoptotic program triggered by NF kappaB selectively in retinal pericytes in response to hyperglycemia is a possible mechanism for the early demise of pericytes in diabetic retinopathy. PMID- 12086957 TI - Elevated spinal cyclooxygenase and prostaglandin release during hyperalgesia in diabetic rats. AB - Diabetic rats display exaggerated hyperalgesic behavior in response to noxious stimuli that may model aspects of painful diabetic neuropathy. This study examined the contribution of spinal prostaglandin production to this exaggerated hyperalgesic behavior. Rats were implanted with spinal dialysis probes and received noxious stimulation to the hind paw by subcutaneous injection of 0.5% formalin solution. Prostaglandin E(2) (PGE(2)) was measured in dialysates of lumbar spinal cerebrospinal fluid concurrent with behavioral responses to formalin injection. In separate experiments, formalin-evoked behavioral responses were measured after intrathecal delivery of either a cyclooxygenase inhibitor or an EP(1) receptor antagonist, and cyclooxygenase protein was measured in spinal cord homogenates. Diabetic rats exhibited exaggerated behavioral responses to paw formalin injection and a concurrent prolongation of formalin-evoked PGE(2) release. Formalin-evoked behavioral responses were dose-dependently reduced in diabetic rats by spinal delivery of a cyclooxygenase inhibitor or an EP(1) receptor antagonist. Protein levels of cyclooxygenase-2 were elevated in the spinal cord of diabetic rats, whereas cyclooxygenase-1 protein was reduced. Hyperalgesic behavior in diabetic rats is associated with both increased cyclooxygenase-2 protein and cyclooxygenase-mediated PGE(2) release. Spinal delivery of selective inhibitors of cyclooxygenase-2 or antagonists of prostaglandin receptors may have therapeutic potential for treating painful diabetic neuropathy. PMID- 12086958 TI - Negative regulation of rho signaling by insulin and its impact on actin cytoskeleton organization in vascular smooth muscle cells: role of nitric oxide and cyclic guanosine monophosphate signaling pathways. AB - Recent studies from our laboratory have shown that insulin induces relaxation of vascular smooth muscle cells (VSMCs) via stimulation of myosin phosphatase and inhibition of Rho kinase activity. In this study, we examined the mechanism whereby insulin inhibits Rho signaling and its impact on actin cytoskeleton organization. Incubation of confluent serum-starved VSMCs with thrombin or phenylephrine (PE) caused a rapid increase in glutathione S-transferase-Rhotekin Rho binding domain-associated RhoA, Rho kinase activation, and actin cytoskeleton organization, which was blocked by preincubation with insulin. Preexposure to N(G)-monomethyl L-arginine acetate (L-NMMA), a nitric oxide synthase inhibitor, and Rp-8 CPT-cyclic guanosine monophosphate (RpcGMP), a cyclic guanosine monophosphate (cGMP) antagonist, attenuated the inhibitory effect of insulin on RhoA activation and restored thrombin-induced Rho kinase activation, and site specific phosphorylation of the myosin-bound regulatory subunit (MBS(Thr695)) of myosin-bound phosphatase (MBP), and caused actin fiber reorganization. In contrast, 8-bromo-cGMP, a cGMP agonist, mimicked the inhibitory effects of insulin and abolished thrombin-mediated Rho activation. Insulin inactivation of RhoA was accompanied by inhibition of isoprenylation via reductions in geranylgeranyl transferase-1 activity as well as increased RhoA phosphorylation, which was reversed by pretreatment with RpcGMP and L-NMMA. We conclude that insulin may inhibit Rho signaling by affecting posttranslational modification of RhoA via nitric oxide/cGMP signaling pathway to cause MBP activation, actin cytoskeletal disorganization, and vasodilation. PMID- 12086959 TI - Minor effect of GLUT1 polymorphisms on susceptibility to diabetic nephropathy in type 1 diabetes. AB - Elevation of intracellular glucose in mesangial cells as mediated by GLUT1 may be important in initiating cellular mechanisms that cause diabetic nephropathy. To determine whether DNA sequence differences in GLUT1 confer susceptibility to this complication, single-nucleotide polymorphisms (SNPs) in this gene were examined using a large case-control study. SNPs examined included the known XbaI (intron 2) and HaeIII SNPs (exon 2). Four novel SNPs located in three putative enhancers were also investigated. Homozygosity for the XbaI(-) allele was associated with diabetic nephropathy (odds ratio 1.83 [95% CI 1.01-3.33]). Furthermore, homozygosity for the A allele for a novel SNP (enhancer-2 SNP 1) located in a putative insulin-responsive enhancer-2 was associated with diabetic nephropathy (2.38 [1.16-4.90]). Patients who were homozygous for risk alleles at both XbaI SNP and enhancer-2 SNP 1 [i.e., homozygosity for XbaI(-)/A haplotype] also had an increased risk of diabetic nephropathy (2.40 [1.13-5.07]). Because enhancer-2 SNP 1 may directly control GLUT1 expression, the strong linkage disequilibrium between the two SNPs likely accounts for XbaI SNP being associated with diabetic nephropathy. In conclusion, our study confirms that SNPs at the GLUT1 locus are associated with susceptibility to diabetic nephropathy in type 1 diabetes. Although these SNPs confer a considerable personal risk for diabetic nephropathy, they account for a limited proportion of cases among type 1 diabetic patients. PMID- 12086960 TI - Modulation of growth hormone signal transduction in kidneys of streptozotocin induced diabetic animals: effect of a growth hormone receptor antagonist. AB - Growth hormone (GH) and IGFs have a long distinguished history in diabetes, with possible participation in the development of renal complications. The implicated effect of GH in diabetic end-stage organ damage may be mediated by growth hormone receptor (GHR) or postreceptor events in GH signal transduction. The present study investigates the effects of diabetes induced by streptozotocin (STZ) on renal GH signaling. Our results demonstrate that JAK2, insulin receptor substrate (IRS)-1, Shc, ERKs, and Akt are widely distributed in the kidney, and after GH treatment, there is a significant increase in phosphorylation of these proteins in STZ-induced diabetic rats compared with controls. Moreover, the GH-induced association of IRS-1/phosphatidylinositol 3-kinase, IRS-1/growth factor receptor bound 2 (Grb2), and Shc/Grb2 are increased in diabetic rats as well. Immunohistochemical studies show that GH-induced p-Akt and p-ERK activation is apparently more pronounced in the kidneys of diabetic rats. Administration of G120K-PEG, a GH antagonist, in diabetic mice shows inhibitory effects on diabetic renal enlargement and reverses the alterations in GH signal transduction observed in diabetic animals. The present study demonstrates a role for GH signaling in the pathogenesis of early diabetic renal changes and suggests that specific GHR blockade may present a new concept in the treatment of diabetic kidney disease. PMID- 12086961 TI - Endothelial dysfunction relates to folate status in children and adolescents with type 1 diabetes. AB - Endothelial dysfunction occurs early in the development of vascular disease in diabetes. Total plasma homocyst(e)ine (tHcy) is associated with endothelial dysfunction. We therefore aimed to assess endothelial function in children with type 1 diabetes in relation to tHcy and its determinants. Endothelial function was assessed in 36 children with type 1 diabetes aged 13.7 +/- 2.2 years and 20 age- and sex-matched control subjects using ultrasound assessment of flow mediated dilatation (FMD) and glyceryl trinitrate (GTN)-dependent brachial artery responses. von Willebrand factor (vWF) and thrombomodulin, markers of endothelial activation, were measured in 64 children with type 1 diabetes and 52 control subjects. Fasting glucose, tHcy, serum and red cell folate, vitamin B12, HbA(1c), creatinine, and lipids were also measured. FMD (5.2 +/- 4.7 vs. 9.1 +/- 4.0%, P = 0.002) and the ratio of FMD:GTN-induced dilatation (0.22 +/- 0.39 vs. 0.41 +/- 0.29%, P = 0.008) were significantly lower in diabetic subjects, indicating endothelial dysfunction. In diabetic subjects, red cell folate correlated independently with FMD (beta = 0.42, P = 0.028) and the ratio of FMD:GTN-induced dilatation (beta = 0.59, P < 0.001). Resting vessel diameter correlated independently with tHcy (beta = -0.51, P < 0.001) and height (beta = 0.65, P < 0.001). vWF correlated independently with HbA(1c) (beta = 0.38, P = 0.003), and thrombomodulin correlated independently with red cell folate (beta = -0.38, P = 0.005), tHcy (beta = -0.37, P = 0.004), diastolic blood pressure (beta = -0.28, P = 0.025), and creatinine clearance (beta = 0.26, P = 0.033). Children with type 1 diabetes have early endothelial dysfunction. Better folate status is associated with better endothelial function, as measured by higher FMD, higher FMD:GTN ratio, and lower thrombomodulin. Folate may therefore protect against endothelial dysfunction in children with diabetes. PMID- 12086962 TI - Hyperketonemia increases tumor necrosis factor-alpha secretion in cultured U937 monocytes and Type 1 diabetic patients and is apparently mediated by oxidative stress and cAMP deficiency. AB - An elevated blood level of tumor necrosis factor (TNF)-alpha is a validated marker of vascular inflammation, which can result in the development of vascular disease and atherosclerosis. This study examined the hypothesis that ketosis increases the TNF-alpha secretion, both in a cell culture model using U937 monocytes and in type 1 diabetic patients in vivo. U937 cells were cultured with ketone bodies (acetoacetate [AA] and beta-hydroxybutyrate [BHB]) in the presence or absence of high levels of glucose in medium at 37 degrees C for 24 h. This study demonstrates the following points. First, hyperketonemic diabetic patients have significantly higher levels of TNF-alpha than normoketonemic diabetic patients (P < 0.01) and normal control subjects (P < 0.01). There was a significant correlation (r = 0.36, P < 0.05; n = 34) between ketosis and oxidative stress as well as between oxidative stress and TNF-alpha levels (r = 0.47, P < 0.02; n = 34) in the blood of diabetic patients. Second, ketone body AA treatment increases TNF-alpha secretion, increases oxygen radicals production, and lowers cAMP levels in U937 cells. However, BHB did not have any effect on TNF alpha secretion or oxygen radicals production in U937 cells. Third, exogenous addition of dibutyryl cAMP, endogenous stimulation of cAMP production by forskolin, and antioxidant N-acetylcysteine (NAC) prevented stimulation of TNF alpha secretion caused by AA alone or with high glucose. Similarly, NAC prevented the elevation of TNF-alpha secretion and lowering of cAMP levels in H(2)O(2) treated U937 cells. Fourth, the effect of AA on TNF-alpha secretion was inhibited by specific inhibitors of protein kinase A (H89), p38-mitogen-activated protein kinase (SB203580), and nuclear transcription factor (NF)kappaB (NFkappaB-SN50). This study demonstrates that hyperketonemia increases TNF-alpha secretion in cultured U937 monocytic cells and TNF-alpha levels in the blood of type 1 diabetic patients and is apparently mediated by AA-induced cellular oxidative stress and cAMP deficiency. PMID- 12086963 TI - Vitamin D receptor (VDR) mRNA and VDR protein levels in relation to vitamin D status, insulin secretory capacity, and VDR genotype in Bangladeshi Asians. AB - Associations have been reported between vitamin D receptor (VDR) gene polymorphisms, type 1 diabetes, insulin secretion, and the insulin resistance syndrome. As VDR polymorphisms have no known functional significance, these findings may implicate a variant of the VDR gene or a locus in linkage disequilibrium with the VDR. We have examined VDR mRNA and VDR protein levels in relation to VDR polymorphisms (41 Bangladeshi subjects) and analyzed insulin secretory capacity (143 Bangladeshi subjects), allowing for other known determinants. Peripheral blood mononuclear cells (PBMCs) from subjects who had been genotyped for BsmI, ApaI, TaqI, and FokI VDR restriction fragment length polymorphisms were used for both total VDR mRNA quantitation (using TaqMan) and measurement of VDR protein levels (using a specific micro-immunoassay). Stepwise multiple regression analyses were used (to P < 0.05) to analyze the data. For the insulin secretion index, the best-fit model (n = 143, P < 0.0001) gave age (P = 0.002), TaqI (P < 0.0001), and BMI (P = 0.001) as independent determinants; with the inclusion of VDR mRNA and VDR protein levels, VDR mRNA was the sole independent determinant (n = 41, P = 0.024). However, the best-fit model for VDR mRNA (P = 0.004) gave FokI (P = 0.044) and TaqI (P = 0.04) genotypes and insulin secretory capacity (P = 0.042) as independent determinants. For VDR protein levels, the best-fit model (P = 0.006) gave TaqI genotype (P = 0.005) and circulating 1,25-dihydroxyvitamin-D levels (P = 0.03) as independent determinants. In conclusion, these studies confirm an association between VDR polymorphisms and insulin secretory capacity and demonstrate the VDR genotype to be a significant determinant of VDR mRNA and VDR protein levels in PBMCs, providing functional support to previously described genetic associations with the VDR gene. Furthermore, VDR expression has been shown to be a determinant of insulin secretory capacity. PMID- 12086964 TI - Loss of kinase activity in a patient with Wolcott-Rallison syndrome caused by a novel mutation in the EIF2AK3 gene. AB - Wolcott-Rallison syndrome (WRS) is an autosomal recessive disorder characterized by neonatal or early infancy type 1 diabetes, epiphyseal dysplasia, and growth retardation. Mutations in the EIF2AK3 gene, encoding the eukaryotic initiation factor 2alpha-kinase 3 (EIF2AK3), have been found in WRS patients. Here we describe a girl who came to our attention at 2 months of age with severe hypertonic dehydration and diabetic ketoacidosis. A diagnosis of type 1 diabetes was made and insulin treatment initiated. Growth retardation and microcephaly were also present. Anti-islet cell autoantibodies were negative, and mitochondrial diabetes was excluded. Imaging revealed a hypoplastic pancreas and typical signs of spondylo-epiphyseal dysplasia. The diagnosis of WRS was therefore made at age 5 years. Sequencing analysis of her EIF2AK3 gene revealed the presence of a homozygous T to C exchange in exon 13 leading to the missense serine 877 proline mutation. The mutated kinase, although it partly retains the ability of autophosphorylation, is unable to phosphorylate its natural substrate, eukaryotic initiation factor 2alpha (eIF2alpha). This is the first case in which the pathophysiological role of EIF2AK3 deficiency in WRS is confirmed at the molecular level. Our data demonstrate that EIF2AK3 kinase activity is essential for pancreas islet function and bone development in humans, and we suggest EIF2AK3 as a possible target for therapeutic intervention in diabetes. PMID- 12086965 TI - A haplotype at the adiponectin locus is associated with obesity and other features of the insulin resistance syndrome. AB - Adiponectin is a protein secreted by adipocytes that modulates insulin action. To assess whether variants of this gene contribute to the prevalence of insulin resistance in Caucasians, we genotyped 413 nondiabetic individuals for two single nucleotide polymorphisms (SNPs) at this locus. The two SNPs (45T-->G and 276G- >T) were chosen because of their association with type 2 diabetes in Japanese. Whereas each polymorphism was significantly associated with some correlate of insulin resistance, the haplotype defined by the two together was strongly associated with many components of the insulin resistance syndrome. Homozygotes for the risk haplotype had higher body weight (P = 0.03), waist circumference (P = 0.004), systolic (P = 0.01) and diastolic (P = 0.003) blood pressure, fasting glucose (P = 0.02) and insulin (P = 0.005) levels, homeostasis model assessment (HOMA) for insulin resistance (P = 0.003), and total to HDL cholesterol ratio (P = 0.01). Homozygotes also had significantly lower plasma levels of adiponectin (P = 0.03), independent of sex, age, and body weight. In an independent study group of 614 Caucasians, including 310 with type 2 diabetes, the risk haplotype was confirmed to be associated with increased body weight (P = 0.03) but not with type 2 diabetes per se. We conclude that variability at the adiponectin locus is associated with obesity and other features of the insulin resistance syndrome, but given the nature of the two SNPs, the risk haplotype is most probably a marker in linkage disequilibrium with an as yet unidentified polymorphism that affects plasma adiponectin levels and insulin sensitivity. PMID- 12086966 TI - A putative functional polymorphism in the IGF-I gene: association studies with type 2 diabetes, adult height, glucose tolerance, and fetal growth in U.K. populations. AB - IGF-I has a critical role in growth and metabolism. A microsatellite polymorphism 1 kb upstream to the IGF-I gene has recently been associated with several adult phenotypes. In a large Dutch cohort, the absence of the commonest allele (Z) was associated with reduced serum IGF-I levels, reduced height, and an increased risk of type 2 diabetes and myocardial infarction. This result has not been replicated, and the role of this polymorphism in these traits in U.K. subjects is not known. We sought further evidence for the involvement of this variant in type 2 diabetes using a case-control study and IGF-I and diabetes-related traits in a population cohort of 640 U.K. individuals aged 25 years. Absence of the common allele was not associated with type 2 diabetes (odds ratio 0.70, 95% CI 0.47-1.04 for X/X versus Z/Z genotype, chi(2) test for trend across genotypes, P = 0.018). In the population cohort, the common allele (Z) was associated with decreased IGF I levels (P = 0.01), contrary to the Dutch study, but not with adult height (P = 0.23), glucose tolerance (P = 0.84), oral glucose tolerance test-derived values of beta-cell function (P = 0.90), or insulin resistance (P = 0.66). There was no association with measures of fetal growth, including birth weight (P = 0.17). Our results do not support the previous associations and suggest that the promoter microsatellite is unlikely to be functionally important. PMID- 12086967 TI - Defining the importance of mitochondrial gene defects in maternally inherited diabetes by sequencing the entire mitochondrial genome. AB - For any mitochondrial DNA (mtDNA) mutation, the ratio of mutant to wild-type mtDNA (% heteroplasmy) varies across tissues, with low levels in leukocytes and high levels in postmitotic tissues (e.g., skeletal muscle). Direct sequencing is the gold-standard method used to detect novel mutations, but can only reliably detect % heteroplasmy >25%, which is rare in leukocytes. Therefore, we investigated the role of mtDNA defects in maternally inherited diabetes by first screening for the A3243G tRNA(Leu(UUR)) mutation by restriction assay, followed by sequencing of the entire mitochondrial genome using skeletal muscle derived mtDNA. A total of 28 patients had maternally inherited diabetes either alone (group 1, n = 17) or with one or more additional features of mitochondrial disease, including bilateral sensori-neural deafness and neuromuscular disease (group 2, n = 11). Three patients (all from group 2) carried the A3243G mutation. Skeletal muscle mtDNA from eight group 1 patients and six more group 2 patients was sequenced. No pathogenic mutations were found in the group 1 patients, while two patients from group 2 had mutations at positions 12258 and 14709 in the tRNA serine and glutamic acid genes, respectively. We conclude, therefore, that screening for mtDNA mutations should be considered in patients with maternally inherited diabetes, but only when additional features of mitochondrial disease are present. PMID- 12086968 TI - The effects of the Pro12Ala polymorphism of the peroxisome proliferator-activated receptor-gamma2 gene on insulin sensitivity and insulin metabolism interact with size at birth. AB - Type 2 diabetes is known to be associated with a small body size at birth. Body size at birth is an indicator of the intrauterine environment. There is also a well-established association between the peroxisome proliferator-activated receptor (PPAR)-gamma2 gene and type 2 diabetes. We therefore assessed whether the effects of the Pro12Ala polymorphism of the PPAR-gamma2 gene on insulin sensitivity and insulin concentrations in adult life are modified by size at birth. We found that the effects of the Pro12Pro and Pro12Ala polymorphisms of the PPAR-gamma2 gene in elderly people depended on their body size at birth. The well-known association between small body size at birth and insulin resistance was seen only in individuals with the high-risk Pro12Pro allele. In those who had low birth weight, the Pro12Pro polymorphism of the PPAR-gamma2 gene was associated with increased insulin resistance (P < 0.002) and elevated insulin concentrations (P < 0.003). These interactions between the effects of the Pro12Ala polymorphisms of the PPAR-gamma2 gene on adult traits and the effects of birth weight link two previously unknown associations together within the context of type 2 diabetes. We suggest that these findings reflect gene-environment interaction. PMID- 12086969 TI - Association of adiponectin mutation with type 2 diabetes: a candidate gene for the insulin resistance syndrome. AB - Adiponectin, also referred to as AdipoQ or ACRP30, is a plasma protein produced and secreted exclusively from adipose tissue. The protein contains a collagen like domain and a C1q-like globular domain. A protease-generated globular segment enhances fatty acid oxidation in muscles, thereby modulating lipid and glucose metabolism. Plasma adiponectin levels are inversely correlated with the severity of insulin resistance. A recent genome-wide scan study mapped a susceptibility locus for type 2 diabetes and the metabolic syndrome to chromosome 3q27, where the adiponectin gene is located. Here, we screened Japanese patients with type 2 diabetes and age- and BMI-matched nondiabetic control subjects for mutations in adiponectin gene. We identified four missense mutations (R112C, I164T, R221S, and H241P) in the globular domain. Among these mutations, the frequency of I164T mutation was significantly higher in type 2 diabetic patients than in age- and BMI- matched control subjects (P < 0.01). Furthermore, plasma adiponectin concentrations of subjects carrying I164T mutation were lower than those of subjects without the mutation. All the subjects carrying I164T mutation showed some feature of metabolic syndrome, including hypertension, hyperlipidemia, diabetes, and atherosclerosis. Our findings suggest that I164T mutation is associated with low plasma adiponectin concentration and type 2 diabetes. PMID- 12086970 TI - Maturity-onset diabetes of the young caused by a balanced translocation where the 20q12 break point results in disruption upstream of the coding region of hepatocyte nuclear factor-4alpha (HNF4A) gene. AB - Monogenic human disorders have been used as paradigms for complex genetic disease and as tools for establishing important insights into mechanisms of gene regulation and transcriptional control. Maturity-onset diabetes of the young (MODY) is a monogenic dominantly inherited form of diabetes that is characterized by defective insulin secretion from the pancreatic beta-cells. A wide variety of mutation types in five different genes have been identified that result in this condition. There have been no reports of a chromosome deletion or translocation resulting in MODY. We report a pedigree where MODY cosegregates with a balanced translocation [karyotype 46, XX t(3;20) (p21.2;q12)]. The chromosome 20 break point, 20q12, is within the region of one of the known MODY genes, hepatocyte nuclear factor-4alpha (HNF4A). Fluorescence in situ hybridization analysis demonstrated that the break point does not disrupt the coding region of this gene, but it lies at least 6 kb upstream of the conventional promoter (P1). We propose that this mutation disrupts the spatial relationship between the recently described alternate distal pancreatic promoter (P2) and HNF4A. This is the first case of MODY due to a balanced translocation, and it provides evidence to confirm the crucial role of an upstream regulator of HNF4A gene expression in the beta cell. PMID- 12086971 TI - IL-12p40 is associated with type 1 diabetes in Caucasian-American families. AB - The IL-12p40 locus has recently been shown to be associated with type 1 diabetes (1). Here, we report the identification of novel microsatellite and single nucleotide polymorphisms (SNPs) within the IL-12p40 gene and a significant association between a (ATT)n repeat marker and type 1 diabetes in 364 U.S. Caucasian sib-pair families (P < 0.006). Haplotype analysis using the (ATT)n repeat (D5S2941) and the C1159A SNP at the 3' untranslated region of IL-12p40 showed a significant association (P = 0.02). Expression studies in individuals heterozygous for the C1159A SNP indicated that the expression of the 1159A allele is approximately 50% higher than that of the 1159C allele. These results provide genetic and functional evidence for IL-12p40 as a type 1 diabetes susceptibility gene. PMID- 12086972 TI - Soluble guanylate cyclase: an old therapeutic target re-visited. PMID- 12086973 TI - M(3) muscarinic receptors mediate contraction of human urinary bladder. AB - Since muscarinic receptors appear to be the physiologically most important control system for urinary bladder contraction, we have characterized the receptor subtype mediating contraction in response to the muscarinic agonist carbachol in the human bladder. Experiments were based on four antagonists, the non-selective atropine, the M(1)-selective pirenzepine, the M(2)-selective methoctramine and the M(3)-selective darifenacin. All antagonists yielded Schild plots with a slope close to unity. The order of potency (atropine> or =darifenacin>pirenzepine>methoctramine) as well as the estimated antagonist affinities suggested that contraction of the human bladder occurs predominantly if not exclusively via the M(3) receptor. PMID- 12086974 TI - Involvement of NO in the failure of neutrophil migration in sepsis induced by Staphylococcus aureus. AB - 1. Sepsis induced by S. aureus was used to investigate whether neutrophil migration failure to infectious focus correlates with lethality in Gram-positive bacteria-induced sepsis in mice. 2. By contrast with the sub-lethal (SL-group), the lethal (L-group) intraperitoneal inoculum of S. aureus caused failure of neutrophil migration (92% reduction), high CFU in the exudate, bacteremia and impairment of in vitro neutrophil chemotactic activity. 3. Pre-treatments of L group with adequate doses of aminoguanidine prevented the neutrophil migration failure and improved the survival of the animals (pre-treated: 43%; untreated: 0% survival). Thus, the impairment of neutrophil migration in the L-group appears to be mediated by nitric oxide (NO). 4. The injection of S. aureus SL-inoculum in iNOS deficient (-/-) or aminoguanidine-treated wild-type mice (pre- and post treatment), which did not present neutrophil migration failure, paradoxically caused severe peritonitis and high mortality. This fact is explainable by the lack of NO dependent microbicidal activity in migrated neutrophils. 5. In conclusion, although the NO microbicidal mechanism is active in neutrophils, the failure of their migration to the infectious focus may be responsible for the severity and outcome of sepsis. PMID- 12086975 TI - The anti-epileptic drug levetiracetam reverses the inhibition by negative allosteric modulators of neuronal GABA- and glycine-gated currents. AB - 1. In this study in vitro and in vivo approaches were combined in order to investigate if the anti-epileptic mechanism(s) of action of levetiracetam (LEV; Keppra) may involve modulation of inhibitory neurotransmission. 2. GABA- and glycine-gated currents were studied in vitro using whole-cell patch-clamp techniques applied on cultured cerebellar granule, hippocampal and spinal neurons. Protection against clonic convulsions was assessed in vivo in sound susceptible mice. The effect of LEV was compared with reference anti-epileptic drugs (AEDs): carbamazepine, phenytoin, valproate, clonazepam, phenobarbital and ethosuximide. 3. LEV contrasted the reference AEDs by an absence of any direct effect on glycine-gated currents. At high concentrations, beyond therapeutic relevance, it induced a small reduction in the peak amplitude and a prolongation of the decay phase of GABA-gated currents. A similar action on GABA-elicited currents was observed with the reference AEDs, except ethosuximide. 4. These minor direct effects contrasted with a potent ability of LEV (EC(50)=1 - 10 microM) to reverse the inhibitory effects of the negative allosteric modulators zinc and beta-carbolines on both GABA(A) and glycine receptor-mediated responses. 5. Clonazepam, phenobarbital and valproate showed a similar ability to reverse the inhibition of beta-carbolines on GABA-gated currents. Blockade of zinc inhibition of GABA responses was observed with clonazepam and ethosuximide. Phenytoin was the only AED together with LEV that inhibited the antagonism of zinc on glycine-gated currents and only clonazepam and phenobarbital inhibited the action of DMCM. 6. LEV (17 mg kg(-1)) produced a potent suppression of sound induced clonic convulsions in mice. This protective effect was significantly abolished by co-administration of the beta-carboline FG 7142, from a dose of 5 mg kg(-1). In contrast, the benzodiazepine receptor antagonist flumazenil (up to 10 mg kg(-1)) was without any effect on the protection afforded by LEV. 7. The results of the present study suggest that a novel ability to oppose the action of negative modulators on the two main inhibitory ionotropic receptors may be of relevance for the anti-epileptic mechanism(s) of action of LEV. PMID- 12086977 TI - Elevated mean arterial pressure in the ovariectomized rat was normalized by ET(A) receptor antagonist therapy: absence of cardiac hypertrophy and fibrosis. AB - 1. The influence of menopause on ventricular function and remodelling remains undefined. The following study examined the effect of ovariectomy on ventricular contractility, cardiac hypertrophy and extracellular matrix protein expression. 2. Elevated circulating levels of the vasoconstrictor endothelin-1 have been reported in post-menopausal women. Moreover, endothelin-1 has been shown to influence blood pressure, ventricular function and cardiac remodelling. In this regard, the potential pathophysiological role of endothelin-1 in the ovariectomized rat was assessed via the administration of the selective endothelin(A) receptor (ET(A)) antagonist BMS-182874. 3. In 3 and 6 week ovariectomized female Sprague - Dawley rats, uterus atrophy was associated with a significant increase in mean arterial pressure, and left ventricular systolic pressure, as compared to sham. By contrast, right ventricular contractile indices were normal in the ovariectomized rat. Despite increased systolic load, left ventricular hypertrophy was not evident, prepro-atrial natriuretic peptide (prepro-ANP) mRNA levels and collagen protein content were similar to sham. 4. The treatment of ovariectomized rats with BMS-182874 (60 mg kg(-1) per day) did not reverse uterus atrophy. However, BMS-182874 normalized mean arterial pressure, and left ventricular systolic pressure in the ovariectomized rat. 5. Thus, despite elevated blood pressure, ovariectomized rats were not associated with either cardiac hypertrophy or fibrosis. Lastly, endothelin-1, acting via the stimulation of the ET(A) receptor represents an integral mechanism implicated in the increase of mean arterial pressure following ovariectomy. PMID- 12086976 TI - Pre- and postsynaptic volatile anaesthetic actions on glycinergic transmission to spinal cord motor neurons. AB - 1. A common anaesthetic endpoint, prevention of withdrawal from a noxious stimulus, is determined primarily in spinal cord, where glycine is an important inhibitory transmitter. To define pre- and postsynaptic anaesthetic actions at glycinergic synapses, the effects of volatile anaesthetic agents on spontaneous and evoked glycinergic currents in spinal cord motor neurons from 6 - 14-day old rats was investigated. 2. The volatile anaesthetic agents enflurane, isoflurane and halothane significantly increased the frequency of glycinergic mIPSCs, enflurane to 190.4% of control+/-22.0 (mean+/-s.e.m., n=7, P<0.01), isoflurane to 199.0%+/-28.8 (n=7, P<0.05) and halothane to 198.2%+/-19.5 (n=7, P<0.01). However without TTX, isoflurane and halothane had no significant effect and enflurane decreased sIPSC frequency to 42.5% of control+/-12.4 (n=6, P<0.01). All the anaesthetics prolonged the decay time constant (tau) of both spontaneous and glycine-evoked currents without increasing amplitude. With TTX total charge transfer was increased; without TTX charge transfer was unchanged (isoflurane and halothane) or decreased (enflurane). 3. Enflurane-induced mIPSC frequency increases were not significantly affected by Cd(2+) (50 microM), thapsigargin (1 5 microM), or KB-R7943 (5 microM). KB-R7943 and thapsigargin together abolished the enflurane-induced increase in mIPSC frequency. 4. There are opposing facilitatory and inhibitory actions of volatile anaesthetics on glycine release dependent on calcium homeostatic mechanisms and sodium channels respectively. Under normal conditions (no TTX) the absolute amount of glycinergic inhibition does not increase. The contribution of glycinergic inhibition to anaesthesia may depend on its duration rather than its absolute magnitude. PMID- 12086978 TI - Effects of sildenafil on erectile activity in mice lacking neuronal or endothelial nitric oxide synthase. AB - 1. Using an in vivo model of erectile activity, the effects of sildenafil were studied in mice lacking neuronal or endothelial nitric oxide synthase (nNOS and eNOS, respectively). 2. Under pentobarbitone anaesthesia, intracavernous pressure (ICP) and mean arterial pressure (MAP) were monitored continuously in wild-type, nNOS-/- and eNOS-/- mice. The magnitude of erectile activity was quantified as the ratio of ICP to MAP. 3. No differences in basal ICP or MAP were observed amongst wild-type, eNOS-/- and nNOS-/- mice. Electrical stimulation of the cavernous nerve (ESCN; 4.0 V, 16 Hz, 1 ms, 30 s) evoked increases in ICP and ICP/MAP as well as penile tumescence. Responses to ESCN were reduced in nNOS-/-, but not in eNOS-/- mice. 4. L-NAME (50 mg kg(-1), i.v.) significantly increased MAP and attenuated erectile responses in both wild-type and eNOS-/- mice. 5. Sildenafil (1 mg kg(-1), i.v.) augmented electrically-evoked erectile activity in a voltage-dependent manner in wild-type mice and facilitated erectile responses in eNOS-/- mice. By contrast, sildenafil failed to augment the diminished erectile responses in mice lacking the nNOS isoform. 5. These data reveal the relative importance of nNOS, compared to eNOS, as the critical NOS isoform in the control of erectile function and illustrate that the nNOS isoform is required for sildenafil-induced facilitation of erectile responses in vivo in mice. PMID- 12086979 TI - Enhancement of apomorphine-induced penile erection in the rat by a selective alpha(1D)-adrenoceptor antagonist. AB - 1. Effects of A-322312 (alpha(1B)-adrenoceptor (AR) antagonist), A-119637 (alpha(1D)-AR antagonist), prazosin (non-selective alpha(1)-AR antagonist), and yohimbine (alpha(2)-AR antagonist) were studied in rat corpus cavernosum (CC) and cavernous artery (Acc) preparations. Effects of intracavernous (i.c.) or intraperitoneal (i.p.) administration of alpha(1)-AR antagonists on apomorphine induced erections were investigated. 2. A-119637 attenuated electrically induced contractions in isolated CC (-logIC(50); 8.12+/-0.15), and relaxed noradrenaline (NA)-contracted preparations by more than 90% at 10(-7) M. At the same concentration, the -logEC(50) value for NA in Acc was altered from 6.79+/-0.07 to 4.86+/-0.13. In the CC and Acc, prazosin similarly inhibited contractile responses. 3. Inhibitory effects of A-322312 (10(-7) M) in electrically activated CC were 32.3+/-5.1%, whereas no effect on concentration-response curves for NA was observed in the Acc. Yohimbine (10(-8) M and 10(-7) M), enhanced electrically induced contractions in isolated CC by 20 to 50%. At 10(-6) M, inhibitory effects of yohimbine were obtained. 4. A-119637 (0.3 micromol kg(-1), i.p.) tripled the number of erections, and produced a 6 fold increase in the duration of apomorphine-induced erectile responses. A-322312, prazosin, or yohimbine did not enhance erections induced by apomorphine. None of the alpha(1)-AR antagonists significantly increased ICP upon i.c. administration. Decreases in blood pressure were seen with A-119637 and prazosin. 5. The present findings show that there is a functional predominance of the alpha(1D)-AR subtype in the rat erectile tissue, and that blockade of this receptor facilitates rat penile erection induced by a suboptimal dose of apomorphine. PMID- 12086980 TI - The extracellular regulated kinases (ERK) 1/2 mediate cannabinoid-induced inhibition of gap junctional communication in endothelial cells. AB - 1. Cannabinoids are potent inhibitors of endothelium-derived hyperpolarizing factor (EDHF)-mediated relaxations. We set out to study the mechanism underlying this effect and the possible role of cannabinoid-induced changes in intercellular gap junction communication. 2. In cultured endothelial cells, Delta(9) tetrahydrocannabinol (Delta(9)-THC) and the cannabinoid receptor agonist HU210, increased the phosphorylation of extracellular regulated kinases 1/2 (ERK1/2) and inhibited gap junctional communication, as determined by Lucifer Yellow dye transfer and electrical capacity measurements. 3. Delta(9)-THC elicited a pronounced increase in the phosphorylation of connexin 43, which was sensitive to PD98059 and U0126, two inhibitors of ERK1/2 activation. Inhibition of ERK1/2 also prevented the Delta(9)-THC-induced inhibition of gap junctional communication. 4. Delta(9)-THC prevented both the bradykinin-induced hyperpolarization and the nitric oxide and prostacyclin-independent relaxation of pre-contracted rings of porcine coronary artery. These effects were prevented by PD98059 as well as U0126. 5. In the absence of Delta(9)-THC, neither PD98059 nor U0126 affected the NO-mediated relaxation of coronary artery rings but both substances induced a leftward shift in the concentration - relaxation curve to bradykinin when diclofenac and N(omega)nitro-L-arginine were present. Moreover, PD98059 and U0126 prolonged the bradykinin-induced hyperpolarization of porcine coronary arteries, without affecting the magnitude of the response. 6. These results indicate that the cannabinoid-induced activation of ERK1/2, which leads to the phosphorylation of connexin 43 and inhibition of gap junctional communication, may partially account for the Delta(9)-THC-induced inhibition of EDHF-mediated relaxation. Moreover, the activation of ERK1/2 by endothelial cell agonists such as bradykinin, appears to exert a negative feedback inhibition on EDHF-mediated responses. PMID- 12086981 TI - Inhibition of the current of heterologously expressed HERG potassium channels by flecainide and comparison with quinidine, propafenone and lignocaine. AB - 1. The inhibition of the cardiac 'rapid' delayed rectifier current (I(Kr)) and its cloned equivalent HERG mediate QT interval prolonging effects of a wide range of clinically used drugs. In this study, we investigated the effects of the Class Ic antiarrhythmic agent flecainide (FLEC) on ionic current (I(HERG)) mediated by cloned HERG channels at 37 degrees C. We also compared the inhibitory potency of FLEC with other Class I agents: quinidine (QUIN, Class Ia); lignocaine (LIG, Class Ib) and propafenone (PROPAF, Class Ic). 2. Whole cell voltage clamp recordings of I(HERG) were made from an HEK293 cell line stably expressing HERG. FLEC inhibited I(HERG) 'tails' following test pulses to +30 mV with an IC(50) of 3.91+/-0.68 microM (mean+/-s.e.mean) and a Hill co-efficient close to 1 (0.76+/ 0.09). 3. In experiments in which I(HERG) tails were monitored following voltage commands to a range of test potentials, I(HERG) inhibition by FLEC was observed to be voltage-dependent and to be associated with a approximately -5 mV shift of the activation curve for the current. Voltage-dependence of inhibition was greatest over the range of potentials corresponding to the steep portion of the I(HERG) activation curve. The time-course of I(HERG) tail deactivation was not significantly altered by FLEC. 4. In experiments in which 10 s depolarizing pulses were applied from -80 to 0 mV, the level of current inhibition by FLEC did not increase between 1 and 10 s. Some time-dependence of inhibition was observed during the first 200 - 300 ms of depolarization. This observation and the voltage dependence of inhibition are collectively consistent with FLEC exerting a rapid open channel state inhibition of I(HERG). 5. Under similar recording conditions QUIN inhibited I(HERG) with an IC(50) of 0.41+/-0.04 microM and PROPAF inhibited I(HERG) with an IC(50) of 0.44+/-0.07 microM. Similar to FLEC, both QUIN and PROPAF showed voltage-dependence of inhibition and blockade developed rapidly during a sustained depolarization. 6. LIG showed little effect on I(HERG) at low micromolar concentrations, but could inhibit the current at higher concentrations; the observed IC(50) was 262.90+/-22.40 microM. 7. Our data are consistent with FLEC, PROPAF and QUIN exerting I(HERG) blockade at clinically relevant concentrations. The rank potency as HERG blockers of the Class I drugs tested in this study was QUIN=PROPAF>FLEC>>LIG. PMID- 12086982 TI - Role of adrenergic receptors in vascular remodelling of the rat choroid. AB - 1. Choroidal blood vessels, located between the sclera and retina, constitute the principle source of blood flow to ocular structures. The choroid is innervated by vasoconstrictor sympathetic and vasodilator parasympathetic nerves. 2. We have shown previously that sympathetic denervation for 6 weeks leads to significant increases in choroidal thickness, percentage of choroid occupied by vascular lumina, and numbers of choroidal venules, large arterioles and outer retinal capillaries. Sympathetic deafferentation produces similar increases, indicating that loss of sympathetic nerve activity is responsible for increased vascularity after sympathectomy. Thus, sympathetic neurotransmission normally may be important in suppressing vascular proliferation in the adult rodent eye. 3. The aim of the present study was to determine whether sympathetic nerves act by way of adrenergic receptors to maintain normal choroidal vascular integrity. 4. The alpha-adrenoceptor antagonist, phentolamine (1 mg kg(-1) day(-1)), the beta receptor antagonist, propranolol (1 mg kg(-1) day(-1)), or saline vehicle was infused for 3 weeks using subcutaneously implanted osmotic minipumps. 5. In phentolamine treated rats, no significant changes were noted relative to saline infused controls. However, propranolol treatment resulted in increases in choroidal thickness, vascular luminal area, and numbers of large choroidal venules and both small and large arterioles, approximating the remodelling seen after chronic sympathectomy. 6. We conclude that sympathetic nerves play a role in maintaining normal choroidal vascular architecture through actions mediated primarily by beta-adrenoceptors. PMID- 12086984 TI - Effect of metabolic inhibition on glimepiride block of native and cloned cardiac sarcolemmal K(ATP) channels. AB - 1. We have investigated the effects of the sulphonylurea, glimepiride, currently used to treat type 2 diabetes, on ATP-sensitive K(+) (K(ATP)) currents of rat cardiac myocytes and on their cloned constituents Kir6.2 and SUR2A expressed in HEK 293 cells. 2. Glimepiride blocked pinacidil-activated whole-cell K(ATP) currents of cardiac myocytes with an IC(50) of 6.8 nM, comparable to the potency of glibenclamide in these cells. Glimepiride blocked K(ATP) channels formed by co expression of Kir6.2/SUR2A subunits in HEK 293 cells in outside-out excised patches with a similar IC(50) of 6.2 nM. 3. Glimepiride was much less effective at blocking K(ATP) currents activated by either metabolic inhibition (MI) with CN(-) and iodoacetate or by the K(ATP) channel opener diazoxide in the presence of inhibitors of F(0)/F(1)-ATPase (oligomycin) and creatine kinase (DNFB). Thus 10 microM glimepiride blocked pinacidil-activated currents by >99%, MI-activated currents by 70% and diazoxide-activated currents by 82%. 4. In inside-out patches from HEK 293 cells expressing the cloned K(ATP) channel subunits Kir6.2/SUR2A, increasing the concentration of ADP (1 - 100 microM), in the presence of 100 nM glimepiride, lead to significant increases in Kir6.2/SUR2A channel activity. However, over the range tested, ADP did not affect cloned K(ATP) channel activity in the presence of 100 nM glibenclamide. These results are consistent with the suggestion that ADP reduces glimepiride block of K(ATP) channels. 5. Our results show that glimepiride is a potent blocker of native cardiac K(ATP) channels activated by pinacidil and blocks cloned Kir6.2/SUR2A channels activated by ATP depletion with similar potency. However, glimepiride is much less effective when K(ATP) channels are activated by MI and this may reflect a reduction in glimepiride block by increased intracellular ADP. PMID- 12086983 TI - Airway function, oedema, cell infiltration and nitric oxide generation in conscious ozone-exposed guinea-pigs: effects of dexamethasone and rolipram. AB - 1. The effects of ozone inhalation (90 min, 2.15+/-0.05 p.p.m.) and their modification by dexamethasone (20 mg kg(-1)) or the phosphodiesterase-4 inhibitor, rolipram (1 mg kg(-1)), administered (i.p.) 24 and 0.5 h before and 24 h after ozone exposure were examined in conscious guinea-pigs. 2. Ozone caused an early-phase bronchoconstriction (EPB) as a fall in specific airways conductance (sG(aw)) measured by whole body plethysmography, followed at 5 h by a late-phase bronchoconstriction (LPB) and increased respiratory rate. Rolipram did not alter this profile but dexamethasone inhibited the EPB. 3. Airway hyperreactivity to inhaled histamine (1 mM, 20 s) occurred at 0.5, 2, 12, 24 and 48 h after ozone inhalation, the 2 h change being abolished by rolipram and dexamethasone. 4. Bronchoalveolar lavage fluid (BALF) macrophages, eosinophils and neutrophils were significantly (P<0.05) elevated at 12, 24 and 48 h after ozone exposure, the 48 h influx being significantly attenuated (P<0.05) by rolipram and dexamethasone. 5. BALF nitric oxide (NO) metabolites decreased 0.5 h after ozone exposure by 52%, recovered at 2 h and significantly increased at 12 (101%) and 24 h (127%). The elevated NO was unaffected by rolipram or dexamethasone. 6. Lung oedema, measured from wet/dry weight differences, was significant 12, 24 and 48 h after ozone exposure, the latter being significantly attenuated (P<0.05) by rolipram and dexamethasone. 7. Ozone exposure of guinea-pigs produced features common to COPD. Although rolipram and dexamethasone did not affect the airway function changes, they inhibited the inflammation, airway hyperreactivity and oedema. PMID- 12086985 TI - Tonic inhibitory control exerted by opioid peptides in the paraventricular nuclei of the hypothalamus on regional hemodynamic activity in rats. AB - 1. Systemic and regional cardiovascular changes were measured following bilateral microinjection of specific and selective opioid-receptor antagonists into the paraventricular nuclei of the hypothalamus (PVN) of awake, freely moving rats. 2. PVN microinjection of increasing doses of the specific opioid antagonist naloxone - methiodide (1 - 5.0 nmol), or a selective mu-opioid receptor antagonist, beta funaltrexamine (0.05 - 0.5 nmol), evoked important cardiovascular changes characterized by small and transient increases in heart rate (HR) and mean arterial pressure (MAP), vasoconstriction in renal and superior mesenteric vascular beds and vasodilation in the hindquarter vascular bed. 3. No significant cardiovascular changes were observed following PVN administration of the highly selective delta-opioid-receptor antagonist, ICI 174864 (0.1 - 1 nmol), or the selective kappa-opioid-receptor antagonist, nor-binaltorphine (0.1 - 1 nmol). 4. Most of the cardiovascular responses to naloxone (3 nmol) and beta-funaltrexamine (0.5 nmol) were attenuated or abolished by an i.v. treatment with a specific vasopressin V(1) receptor antagonist. 5. These results suggest that endogenous opioid peptides and mu-type PVN opioid receptors modulate a tonically-active central depressor pathway acting on systemic and regional haemodynamic systems. Part of this influence could involve a tonic inhibition of vasopressin release. PMID- 12086986 TI - Central injections of nocistatin or its C-terminal hexapeptide exert anxiogenic like effect on behaviour of mice in the plus-maze test. AB - . Nocistatin (NST) antagonizes several actions of nociceptin/orphanin FQ (N/OFQ), but acts on distinct receptors. As N/OFQ exerts anxiolytic-like actions in various tests, its behavioural actions in the elevated plus-maze (EPM) test were compared with those of bovine NST. 2. Five minutes after i.c.v. treatment, mice were placed on the EPM for 5 min and entries into and time spent on open and closed arms were recorded alongside other parameters. 3. NST (0.1 - 3 pmol) reduced percentages of entries into (control 39.6+/-3.1%, peak effect at 1 pmol NST 8.5+/-2.9%) and time spent on open arms (control 30.8+/-2.3%, NST 2.7+/ 1.5%). The C-terminal hexapeptide of NST (NST-C6; 0.01 - 10 pmol) closely mimicked these actions of NST, with peak effects at 0.1 pmol. 4. N/OFQ (1 - 100 pmol) increased percentages of entries into (control 38.5+/-3.4%; peak effect at 10 pmol N/OFQ 67.9+/-4.9%) and time spent on open arms (control 32.0+/-3.8%; N/OFQ 74.9+/-5.8%). Closed arm entries, an index of locomotor activity, were unchanged by all peptides. 5. Effects of NST or NST-C6, but not N/OFQ, were still detectable 15 min after injection. Behaviour of animals co-injected with NST (1 pmol) or NST-C6 (0.1 pmol) plus N/OFQ (10 pmol) was indistinguishable from that of controls. 6. These results reveal potent anxiogenic-like actions of NST and NST-C6, and confirm the anxiolytic-like properties of N/OFQ. As NST and N/OFQ both derive from preproN/OF, anxiety may be modulated in opposing directions depending on how this precursor is processed. PMID- 12086987 TI - NO- and haem-independent activation of soluble guanylyl cyclase: molecular basis and cardiovascular implications of a new pharmacological principle. AB - 1. Soluble guanylyl cyclase (sGC) is the only proven receptor for the ubiquitous biological messenger nitric oxide (NO) and is intimately involved in many signal transduction pathways, most notably in regulating vascular tone and platelet function. sGC is a heterodimeric (alpha/ss) protein that converts GTP to cyclic GMP; NO binds to its prosthetic haem group. Here, we report the discovery of a novel sGC activating compound, its interaction with a previously unrecognized regulatory site and its therapeutic implications. 2. Through a high-throughput screen we identified BAY 58-2667, an amino dicarboxylic acid which potently activates sGC in an NO-independent manner. In contrast to NO, YC-1 and BAY 41 2272, the sGC stimulators described recently, BAY 58-2667 activates the enzyme even after it has been oxidized by the sGC inhibitor ODQ or rendered haem deficient. 3. Binding studies with radiolabelled BAY 58-2667 show a high affinity site on the enzyme. 4. Using photoaffinity labelling studies we identified the amino acids 371 (alpha-subunit) and 231 - 310 (ss-subunit) as target regions for BAY 58-2667. 5. sGC activation by BAY 58-2667 results in an antiplatelet activity both in vitro and in vivo and a potent vasorelaxation which is not influenced by nitrate tolerance. 6. BAY 58-2667 shows a potent antihypertensive effect in conscious spontaneously hypertensive rats. In anaesthetized dogs the hemodynamic effects of BAY 58-2667 and GTN are very similar on the arterial and venous system. 7. This novel type of sGC activator is a valuable research tool and may offer a new approach for treating cardiovascular diseases. PMID- 12086988 TI - Comparison of the expression of calcitonin receptor-like receptor (CRLR) and receptor activity modifying proteins (RAMPs) with CGRP and adrenomedullin binding in cell lines. AB - 1. The calcitonin receptor-like receptor (CRLR) and specific receptor activity modifying proteins (RAMPs) together form receptors for calcitonin gene-related peptide (CGRP) and/or adrenomedullin in transfected cells. 2. There is less evidence that innate CGRP and adrenomedullin receptors are formed by CRLR/RAMP combinations. We therefore examined whether CGRP and/or adrenomedullin binding correlated with CRLR and RAMP mRNA expression in human and rat cell lines known to express these receptors. Specific human or rat CRLR antibodies were used to examine the presence of CRLR in these cells. 3. We confirmed CGRP subtype 1 receptor (CGRP(1)) pharmacology in SK-N-MC neuroblastoma cells. L6 myoblast cells expressed both CGRP(1) and adrenomedullin receptors whereas Rat-2 fibroblasts expressed only adrenomedullin receptors. In contrast we could not confirm CGRP(2) receptor pharmacology for Col-29 colonic epithelial cells, which, instead were CGRP(1)-like in this study. 4. L6, SK-N-MC and Col-29 cells expressed mRNA for RAMP1 and RAMP2 but Rat-2 fibroblasts had only RAMP2. No cell line had detectable RAMP3 mRNA. 5. SK-N-MC, Col-29 and Rat-2 fibroblast cells expressed CRLR mRNA. By contrast, CRLR mRNA was undetectable by Northern analysis in one source of L6 cells. Conversely, a different source of L6 cells had mRNA for CRLR. All of the cell lines expressed CRLR protein. Thus, circumstances where CRLR mRNA is apparently absent by Northern analysis do not exclude the presence of this receptor. 6. These data strongly support CRLR, together with appropriate RAMPs as binding sites for CGRP and adrenomedullin in cultured cells. PMID- 12086990 TI - Eicosapentaenoic acid regulates scallop (Placopecten magellanicus) membrane fluidity in response to cold. PMID- 12086989 TI - Oncostatin M stimulates proliferation, induces collagen production and inhibits apoptosis of human lung fibroblasts. AB - 1. Oncostatin M (OSM), a member of the interleukin-6 (IL-6) cytokine family, acts on a variety of cells and elicits diversified biological responses, suggesting potential roles in the regulation of cell survival, differentiation and proliferation. 2. We have examined the effect of OSM on the regulation of human lung fibroblast proliferation, collagen production and spontaneous apoptosis. The proliferative effects of OSM (0.5 - 100 ng ml(-1)) were assessed using a MTS assay as well as [(3)H]-thymidine incorporation and cell counts at 24 and 48 h. Hydroxyproline was measured as an index of procollagen production by high pressure liquid chromotography (HPLC). Apoptosis was determined by annexin staining. 3. OSM enhanced the mitotic activity of lung fibroblasts in a time and dose dependent manner. Maximum proliferation of 57% above control was observed after incubation for 48 h with 2 ng ml(-1) OSM (P<0.05). 4. Incubation with the mitogen activated protein kinase (MAPK) kinase inhibitor, PD98059 or the tyrosine kinase inhibitor, genestein both significantly reduced the mitogenic effect of OSM (P<0.05). 5. In contrast, proliferation in response to OSM was not regulated by induction of cyclo-oxygenase and subsequent prostaglandin E(2) (PGE(2)) release or by IL-6. 6. OSM also stimulated fibroblasts to synthesize pro-collagen by a maximum of 35% above control levels after 48 h (P<0.05). 7. OSM significantly inhibited the spontaneous apoptosis of fibroblasts at 24 and 48 h. 8. These results provide evidence that OSM has pro-fibrotic properties and suggest that it may play a role in normal lung wound repair and fibrosis. PMID- 12086992 TI - Role of specialized microvilli and the fertilization envelope in the spatial positioning of blastomeres in early development of embryos of the starfish Astropecten scoparius. AB - In the eggs of a wide range of animal species, various factors that determine the blastomeres' presumptive fate are known to locate unevenly within the egg. In the embryos of these animals, cleavage occurs not just to increase cell numbers, but also to distribute the factors to the respective blastomeres, resulting in cell specialization at the later stages. In the early cleavage stages, before the establishment of a device such as desmosomes to directly join the blastomeres, some other means is needed to keep the blastomeres together and maintain the relative positions among them. In this study, we found that the embryos of the starfish Astropecten scoparius lack the hyaline layer seen in sea urchin embryos and that blastomeres adhere to the fertilization envelope (FE) via filamentous cellular projections (fixing processes). Electron microscopy revealed the fixing processes to be specialized microvilli formed, after the elevation of the FE, by the elongation of short microvilli that pre-exist in unfertilized eggs. After the first cleavage, the two blastomeres separate from each other and finally attach to the FE. In the subsequent cleavages, the blastomeres undergo repeated cell division without separating from the FE. Between the blastomeres and the FE, only shortened fixing processes were observed. Destruction of the fixing processes caused release of the blastomeres from the FE and disturbance of the relative positions of the blastomeres, resulting in abnormal development of the embryos. These observations suggest that the fixing process is a device to keep the egg placed centrally in the FE up to the first cleavage, and after the first cleavage and beyond to anchor the blastomeres to the FE so that the FE can be used as a scaffold for morphogenesis. Electron microscopy also suggests that the inner layer of the FE, which is derived from the contents of cortical granules, reinforces the adhesion of the fixing processes to the FE. Immuno-electron microscopy, using an antibody against sea urchin hyaline layer, showed that the inner layer of the FE of starfish eggs and the hyaline layer of sea urchin eggs, which are both derived from cortical granules, contain some common elements. PMID- 12086991 TI - Characterization of limb autotomy factor-proecdysis (LAF(pro)), isolated from limb regenerates, that suspends molting in the land crab Gecarcinus lateralis. AB - Molting and limb regeneration are tightly coupled processes, both of which are regulated by ecdysteroid hormone synthesized and secreted by the Y-organs. Regeneration of lost appendages can affect the timing and duration of the proecdysial, or premolt, stage of the molt cycle. Autotomy of all eight walking legs induces precocious molts in various decapod crustacean species. In the land crab Gecarcinus lateralis, autotomy of a partially regenerated limb bud before a critical period during proecdysis (regeneration index <17) delays molting so that a secondary limb bud (2 degrees LB) forms and the animal molts with a complete set of walking legs. It is hypothesized that 2 degrees LBs secrete a factor, termed limb autotomy factor-proecdysis (LAF(pro)), that inhibits molting by suppressing the Y-organs from secreting ecdysone. Molting was induced by autotomy of eight walking legs; autotomy of primary (1 degrees ) LBs reduced the level of ecdysteroid hormone in the hemolymph 73% by one week after limb bud autotomy (LBA). Injection of extracts from 2 degrees LBs, but not 1 degrees LBs, inhibited 1 degrees LB growth in proecdysial animals, thus having the same effect on molting as LBA. The inhibitory activity in 2 degrees LB extracts was stable after boiling in water for 15 min, but was destroyed by boiling 15 min in 0.1 N acetic acid or incubation with proteinase K. These results support the hypothesis that LAF(pro) is a peptide that resembles a molt-inhibiting hormone. PMID- 12086993 TI - Postembryonic eye growth in the seashore isopod Ligia exotica (Crustacea, Isopoda). AB - The eye of Ligia exotica is of the apposition type and has open rhabdoms. The facets are hexagonal, and the dioptric apparatus consists of a flat cornea and a spherical crystalline cone placed in the center of two large cone cells. Each ommatidium has seven regular retinula cells and one eccentric cell; a basement membrane forms the proximal boundary of the retina. With increases in body size from 0.6 to almost 4.0 cm, facet numbers and ommatidial diameters increased from 800 to 1500 and 35 microm to 100 microm, respectively; eye length and width grew from 1.2 to 3.2 and 0.9 to 2.5 mm, respectively; and length of dioptric apparatus and width of retinal layer changed from 70 microm to 180 microm and about 70 microm to 120 microm. Visual angles and interommatidial angles of centrally located ommatidia remained constant at about 30 and 6.9 degrees, respectively. An almost perfect linear relationship was found when eye length was plotted against the product between the square root of the total number of ommatidia and the ommatidial diameter. No difference between males and females was observed in any of the relationships, but the results suggest that, compared with smaller specimens, larger ones possess increased absolute sensitivity in single ommatidia, increased sensitivity to point sources, and overall larger angular visual fields for the eye in its totality. This means that larger individuals of L. exotica (which are also faster) have an advantage over smaller individuals at night, but that smaller individuals may cope better with bright lights. Vision in L. exotica seems useful not only in detecting potential danger, but also in locating and approaching cliffs from a distance of 2-4 m when swimming in seawater. PMID- 12086995 TI - The limitations of self-organization in biological systems. Proceedings of a workshop sponsored by the Center for Advanced Studies in the Space Sciences at the Marine Biological Laboratory. Woods Hole, Massachusetts, USA. May 11-13, 2001. PMID- 12086996 TI - Self-organization in relation to several similar concepts: are the boundaries to self-organization indistinct? AB - Self-organization is a concept and phenomenon whereby system-level patterns spontaneously arise solely from interactions among subunits of the system. Focusing on self-organization at the organismal level, I ask the question: are the boundaries to self-organization indistinct? After reviewing a number of published definitions of self-organization, I explore the conceptual boundaries among self-organization and two similar concepts, stigmergy and self-assembly. I highlight borderline cases that may blur the distinction among these and suggest that they may indeed be conceptually indistinct and difficult to separate in practice. Consequently, I propose a classification scheme based upon three aspects: whether the stimuli to which individuals respond are quantitative or qualitative, whether positive feedback is involved, and whether interindividual interactions are direct or indirect (stigmergic). In addition, I consider several other issues about self-organization, including (1) could a self-organized system use global information? (2) what is the role of the degree of correlation of activity among individuals? and (3) what is the role of positive feedback? PMID- 12086994 TI - Trans-atlantic distribution of a mangrove oyster species revealed by 16S mtDNA and karyological analyses. AB - Three species of mangrove oysters, Crassostrea rhizophorae, C. brasiliana, and C. gasar, have been described along the Atlantic shores of South America and Africa. Because the distribution of these molluscs is of great biological and commercial interest, their taxonomy and distribution deserve further clarification. Therefore, 15 populations were sampled from both continents. Their 16S mitochondrial polymorphism was studied by sequencing and PCR-RFLP analysis. Two haplotypes were identified. Haplotype a was the only one observed in Africa, but it was also observed in South America together with haplotype b. Because C. gasar is the only mangrove oyster identified on the west coast of Africa, haplotype a was attributed to this species, which has thus been shown to occur in South America. Haplotype b is attributed to C. rhizophorae. The karyotypes of specimens of C. gasar, from Africa and from South America, were very similar, and both species were observed at the same location in Brazil. The occurrence of C. gasar in South America adds a third species-in addition to C. rhizophorae and C. brasiliana-to the list of species present along these coasts. The predominant surface circulation patterns in this part of the Atlantic Ocean favor the hypothesis that C. gasar was transported from Africa to America. Finally, a phylogenetic tree built with seven 16S sequences from Crassostrea and Saccostrea species showed that C. gasar is intermediate between the American Crassostrea species (C. virginica and C. rhizophorae) and the Asian species (C. gigas and C. ariakensis). PMID- 12086997 TI - Evolution of self-organized systems. AB - In this paper I ask questions about the evolution of self-organized activity cycles that are found in some ant colonies. I use a computer model that generates periodic activity patterns in interacting subunits and explore the parameters of this model using a genetic algorithm in which selecting on one aspect of the system produces the distinctive self-organized pattern. The general point that I explore, using the example of activity cycles, is that the observation of a self organized pattern does not mean that the pattern is an adaptation. Self-organized patterns can represent nonadaptive correlated responses to selection, exaptations or even selectively disadvantageous traits. Evolution of self-organized patterns requires genetic feedback between the self-organized output and the subunits that produce the pattern. Without this necessary feedback, a self-organized system does not evolve. PMID- 12086998 TI - Dynamics of aggregation and emergence of cooperation. AB - Aggregation is one of the most basic social phenomena, and many activities of social insects are linked to it. For instance, the selection of a valuable site and the spatial organization of the population are very often by-products of amplifications based on the local density of nestmates. The patterns of aggregation are very diverse, ranging from the gathering of all animals in a unique site to their splitting between several ones. One might question how these multiple patterns emerge. Do ants actively initiate the formation of such patterns by modulating the emission of an attracting signal such as the trail pheromone? Alternatively, do patterns result from quantitative changes in the duration of interaction between animals once they have reached the gathering site, without any active modulation of the communications? To discuss these questions, we present two empirical studies: the gregarious behavior of cockroaches (Blatella) and self-assembly in the weaver ant (Oecophylla). Through experimental and theoretical studies, we show how a single behavior-the resting time-leads to a collective choice in both species. This behavior is a response to the density of conspecifics and can also be modulated by heterogeneities in the environment. In weaver ants, it allows the colony to focus the formation of chains in a given area among several potential sites. In cockroaches, it allows the gathering of individuals in particular shelters, depending on the proximity between strains. These results are discussed with emphasis on the role of aggregation processes in the emergence of cooperativity and task allocation. PMID- 12086999 TI - Complexity of environment and parsimony of decision rules in insect societies. AB - This paper shows how colonies of social insects process information and solve problems in a complex environment, while keeping some parsimony at the level of the individuals' decision rules. Two studies on ant foraging reveal the diversity of adaptive colony-level patterns that can be generated through self organization, based on the same individual-level recruitment rules. Regarding prey scavenging, the "ability to retrieve the prey" rule accounts for changes in foraging patterns, with increasing prey size, that show all stages intermediate between an individual and a mass exploitation of food resources. Regarding liquid food foraging, the "ability to ingest a desired volume" rule enables a colony to adjust the number of tending ants to the honeydew production of aphids. In both cases, decision rules are based on intelligent criteria that intrinsically integrate information on multiple variables that are relevant to the ants. Furthermore, the environment can contribute directly to the emergence of collective patterns, independently of any individual behavioral changes. Each environmental factor, including abiotic ones, that alters the dynamics of information transfer in group-living animals should be reconsidered not simply as a constraint but also as a part of the decision-making process and as a agent that shapes the collective pattern. PMID- 12087000 TI - Herd size in large herbivores: encoded in the individual or emergent? AB - In large mammalian herbivores, the increase of group size with habitat openness was first assumed to be an adaptive response, encoded in the individual. However, it could, alternatively, be an emergent property: if groups were nonpermanent units, often fusing and splitting up, then any increase of the distance at which animals perceive one another could increase the rate of group fusion and thus mean group size. Dynamical models and empirical data support this second hypothesis. This is not to say that adaptive modifications of mean herd size cannot occur. However, this changes the way in which we can envisage the history of gregariousness in large herbivores during the Tertiary. PMID- 12087001 TI - Self-organization and natural selection in the evolution of complex despotic societies. AB - Differences between related species are usually explained as separate adaptations produced by individual selection. I discuss in this paper how related species, which differ in many respects, may evolve by a combination of individual selection, self-organization, and group-selection, requiring an evolutionary adaptation of only a single trait. In line with the supposed evolution of despotic species of macaques, we take as a starting point an ancestral species that is egalitarian and mildly aggressive. We suppose it to live in an environment with abundant food and we put the case that, if food becomes scarce and more clumped, natural selection at the level of the individual will favor individuals with a more intense aggression (implying, for instance, biting and fierce fighting). Using an individual-centered model, called DomWorld, I show what happens when the intensity of aggression increases. In DomWorld, group life is represented by artificial individuals that live in a homogeneous world. Individuals are extremely simple: all they do is flock together and, upon meeting one another, they may perform dominance interactions in which the effects of winning and losing are self-reinforcing. When the intensity of aggression in the model is increased, a complex feedback between the hierarchy and spatial structure results; via self-organization, this feedback causes the egalitarian society to change into a despotic one. The many differences between the two types of artificial society closely correspond to those between despotic and egalitarian macaques in the real world. Given that, in the model, the organization changes as a side effect of the change of one single trait proper to an egalitarian society, in the real world a despotic society may also have arisen as a side effect of the mutation of a single trait of an egalitarian species. If groups with different intensities of aggression evolve in this way, they will also have different gradients of hierarchy. When food is scarce, groups with the steepest hierarchy may have the best chance to survive, because at least a small number of individuals in such a group may succeed in producing offspring, whereas in egalitarian societies every individual is at risk of being insufficiently fed to reproduce. Therefore, intrademic group selection (selection within an interbreeding group) may have contributed to the evolution of despotic societies. PMID- 12087002 TI - Scaling in nests of a social wasp: a property of the social group. AB - The numbers of brood cells in nests built by founding swarms of the Neotropical social wasp Polybia occidentalis closely correlate with the numbers of wasps in the swarms. We analyzed nests of different sizes to determine how they scale with respect to the allocation of brood cells among combs. Three patterns were evident: compared to smaller nests, larger nests have (1) more combs and (2) larger combs; and (3) among nests containing the same number of combs, the last two combs diverge in relative size as nest size increases. Taken together, these results suggest that members of a swarm somehow "know" the size of the swarm they are in. This information feeds back to individual builders, which quantitatively modulate their responses to stigmergic cues in ways that result in the nest-size scaled allocation of brood cells among combs. The patterns also suggest that swarms fine-tune the final size of their nests by making corrections as they build. PMID- 12087003 TI - Self-organized fish schools: an examination of emergent properties. AB - Heterogeneous, "aggregated" patterns in the spatial distributions of individuals are almost universal across living organisms, from bacteria to higher vertebrates. Whereas specific features of aggregations are often visually striking to human eyes, a heuristic analysis based on human vision is usually not sufficient to answer fundamental questions about how and why organisms aggregate. What are the individual-level behavioral traits that give rise to these features? When qualitatively similar spatial patterns arise from purely physical mechanisms, are these patterns in organisms biologically significant, or are they simply epiphenomena that are likely characteristics of any set of interacting autonomous individuals? If specific features of spatial aggregations do confer advantages or disadvantages in the fitness of group members, how has evolution operated to shape individual behavior in balancing costs and benefits at the individual and group levels? Mathematical models of social behaviors such as schooling in fishes provide a promising avenue to address some of these questions. However, the literature on schooling models has lacked a common framework to objectively and quantitatively characterize relationships between individual-level behaviors and group-level patterns. In this paper, we briefly survey similarities and differences in behavioral algorithms and aggregation statistics among existing schooling models. We present preliminary results of our efforts to develop a modeling framework that synthesizes much of this previous work, and to identify relationships between behavioral parameters and group-level statistics. PMID- 12087004 TI - Individual complexity and self-organization in foraging by leaf-cutting ants. AB - Leaf-cutting ants cut vegetation into small fragments that they transport to the nest, where a symbiotic fungus cultivated by the ants processes the material. Since the harvested leaf fragments are incorporated into the fungus garden and not directly consumed by the workers, it is expected that foraging workers select plants by responding to those physical or chemical traits that promote maximal fungal growth, irrespective of the potential direct effects of these leaf features on them. In this paper I summarize experimental work focusing on the decision-making processes that occur at the individual level, and discuss to what extent individual complexity contributes to the emergence of collective foraging patterns. Although some basic features of self-organizing systems, such as the existence of regulatory positive and negative feedback loops, are expected to be involved in the collective organization of leaf-cutting ant foraging, I contend that they are combined with complex individual responses that may result from the integration of local information during food collection with an assessment of colony conditions. PMID- 12087005 TI - When is self-organization used in biological systems? AB - Self-organization, or decentralized control, is widespread in biological systems, including cells, organisms, and groups. It is not, however, the universal means of organization. I argue that a biological system will be self-organized when it possesses a large number of subunits, and these subunits lack either the communicational abilities or the computational abilities, or both, that are needed to implement centralized control. Such control requires a well informed and highly intelligent supervisor. I stress that the subunits in a self-organized system do not necessarily have low cognitive abilities. A lack of preadaptations for evolving a system-wide communication network can prevent the evolution of centralized control. Hence, sometimes even systems whose subunits possess high cognitive abilities will be self-organized. PMID- 12087006 TI - Elimination of dietary gluten does not reduce titers of type 1 diabetes associated autoantibodies in high-risk subjects. AB - OBJECTIVE: Removal of the dietary wheat protein gluten protects against autoimmune diabetes in animal models. Furthermore, elimination of dietary gluten reduces the frequency of type 1 diabetes in patients with celiac disease. Herein we test the hypothesis that gluten is the driving antigen for type 1 diabetes associated islet autoimmunity. RESEARCH DESIGN AND METHODS: Seven autoantibody positive, first-degree relatives of patients with type 1 diabetes were placed on a gluten-free diet for 12 months followed by gluten reexposure for 12 months. Gliadin antibodies as well as the diabetes-related antibodies insulin autoantibody (IAA), GAD antibody (GADA), and tyrosin phosphatase IA2 antibody (IA 2A) were measured every 3 months; oral glucose tolerance tests were performed every 6 months. Changes in autoantibody titers were compared with those observed in a matched historical cohort. RESULTS: A reduction in IgG gliadin antibody titers was observed during the gluten-free period, but titers of diabetes associated autoantibodies changed independently of gluten exposure. Type 1 diabetes-associated islet autoantibody levels at the end of the gluten-free diet period were not significantly different from those before commencement of the diet (P = 0.2) or at the end of the gluten reexposure period (P = 0.4). Changes in individual subjects were identified, but no differences were noted between the gluten-free and the gluten re-exposure periods, and the changes were similar to those observed in the historical control cohort (P = 1.0). Major titer reductions (>50%) in the gluten-free period were observed in only one subject for all antibodies. Type 1 diabetes developed in this subject and in a second subject during the gluten reexposure period. CONCLUSIONS: The findings do not support the hypothesis that gluten is a driving antigen in type 1 diabetes. PMID- 12087007 TI - A longitudinal study of the effects of a gluten-free diet on glycemic control and weight gain in subjects with type 1 diabetes and celiac disease. AB - OBJECTIVE: To describe the longitudinal growth characteristics and glycemic control in type 1 diabetic children diagnosed with celiac disease and started on a gluten-free diet (GFD). RESEARCH DESIGN AND METHODS: Data on growth and glycemic control for 11 case subjects diagnosed with celiac disease (cd(+) group) and started on a GFD were collected prospectively, and two control subjects without celiac disease matched for age, sex, and duration of diabetes (cd(-) group) were selected for comparison. RESULTS: In the period between diagnosis of type 1 diabetes and start of a GFD in the cd(+) compared with the cd(-) group, BMI standard deviation score (SDS) was lower (-0.2 vs. 0.7, P = 0.015), as was HbA(1c) (8.9 vs. 9.8%, P = 0.002). In a regression model the cd(+) group had lower BMI SDS (P < 0.001) and lower HbA(1c) (P = 0.04), independent of other variables. On a GFD, BMI SDS increased by 12 months in the cd(+) group and then was no different than the cd(-) group (1.1 vs. 1.0, P = 0.11), whereas HbA(1c) improved further within case subjects compared with pre-GFD (8.9 vs. 8.3%, P = 0.002). On a GFD, case subjects in contrast to control subjects showed no deterioration in HbA(1c) during the years of puberty (8.3 vs. 10.0%, P = 0.022) CONCLUSIONS: In children with type 1 diabetes, untreated celiac disease resulted in lower BMI SDS and lower HbA(1c). Recovery of BMI SDS with a GFD was associated with further improvement in HbA(1c) as compared with pre-GFD, with no expected deterioration in glycemic control during puberty. These apparent clinical benefits need confirming by larger studies. PMID- 12087008 TI - Effect of orlistat in overweight and obese patients with type 2 diabetes treated with metformin. AB - OBJECTIVE: The purpose of this study was to assess the effect of orlistat, a gastrointestinal lipase inhibitor, on body weight, glycemic control, and cardiovascular risk factors in metformin-treated type 2 diabetic patients. RESEARCH DESIGN AND METHODS: A 1-year multicenter, randomized, double-blind, placebo-controlled trial of 120 mg orlistat t.i.d. (n = 249) or placebo (n = 254) combined with a reduced-calorie diet was conducted in overweight and obese patients with suboptimal control of type 2 diabetes. RESULTS: After 1 year of treatment, mean (+/-SE) weight loss was greater in the orlistat than in the placebo group (-4.6 +/- 0.3% vs. -1.7 +/- 0.3% of baseline wt, P < 0.001). Orlistat treatment caused a greater improvement in glycemic control than placebo, as evidenced by a greater reduction in serum HbA(1c), adjusted for changes in metformin and sulfonylurea therapy (-0.90 +/- 0.08 vs. -0.61 +/- 0.08, P = 0.014); a greater proportion of patients achieving decreases in HbA(1c) of > or = 0.5 and > or = 1.0% (both P < 0.01); and a greater reduction in fasting serum glucose (-2.0 +/- 0.2 vs. -0.7 +/- 0.2 mmol/l, P = 0.001). Compared with the placebo group, patients treated with orlistat also had greater decreases in total cholesterol, LDL cholesterol, and systolic blood pressure (all P < 0.05). Although more subjects treated with orlistat experienced gastrointestinal side effects than placebo (83 vs. 62%, P < 0.05), more subjects in the placebo group withdrew prematurely from the study than in the orlistat group (44 vs. 35%, P < 0.05). CONCLUSIONS: Orlistat is a useful adjunctive treatment for producing weight loss and improving glycemic control, serum lipid levels, and blood pressure in obese patients with type 2 diabetes who are being treated with metformin. PMID- 12087009 TI - Elevated risk of cardiovascular disease prior to clinical diagnosis of type 2 diabetes. AB - OBJECTIVE: To examine whether the risk of cardiovascular disease (CVD) is elevated before clinical diagnosis of type 2 diabetes in women. RESEARCH DESIGN AND METHODS: A total of 117,629 female nurses aged 30-55 years who were free of diagnosed CVD at baseline were recruited in 1976 and followed for 20 years. RESULTS: A total of 1,508 women had diagnosed type 2 diabetes at baseline in 1976. During 20 years of follow-up, 110,227 women remained free of diabetes diagnosis and 5,894 women developed type 2 diabetes. During 2.2 million person years of follow-up, we documented 1,556 new cases of myocardial infarction (MI), 1,405 strokes, 815 fatal coronary heart disease (CHD), and 300 fatal strokes. Among women who developed type 2 diabetes during follow-up, the age-adjusted RRs of MI were 3.75 (95% CI 3.10-4.53) for the period before the diagnosis and 4.57 (3.87-5.39) for the period after the diagnosis, compared with women who remained free of diabetes diagnosis. The multivariate RRs further adjusting for BMI, smoking, and other cardiovascular risk factors were 3.17 (2.61-3.85) and 3.97 (3.35-4.71). The risk of stroke was also significantly elevated before diagnosis of diabetes (multivariate RR = 2.30 [1.76-2.99]). Further adjustment for history of hypertension or hypercholesterolemia did not appreciably alter the results. CONCLUSIONS: Our data indicate a substantially elevated risk of CVD before clinical diagnosis of type 2 diabetes in women. These findings suggest that aggressive management of cardiovascular risk factors is warranted in individuals at increased risk for diabetes. PMID- 12087010 TI - HOMA-estimated insulin resistance is an independent predictor of cardiovascular disease in type 2 diabetic subjects: prospective data from the Verona Diabetes Complications Study. AB - OBJECTIVE: To evaluate whether homeostasis model assessment-estimated insulin resistance (HOMA-IR) is an independent predictor of cardiovascular disease (CVD) in type 2 diabetes. RESEARCH DESIGN AND METHODS: Conventional CVD risk factors (sex, age, smoking, plasma lipids, blood pressure, and metabolic control) and insulin resistance (estimated by HOMA) were evaluated at baseline in 1,326 patients with type 2 diabetes examined within the Verona Diabetes Complications Study. At baseline and after a mean follow-up of 4.5 years, CVD was assessed by medical history, physical examination, electrocardiography, and echo-Doppler of carotid and lower limb arteries. Death certificates and medical records of subjects who died during the follow-up were carefully scrutinized to identify cardiovascular deaths. In statistical analyses, CVD was an aggregate end point including both fatal and nonfatal coronary, cerebrovascular, and peripheral vascular disease as well as ischemic electrocardiographic abnormalities and vascular lesions identified by echo-Doppler. RESULTS: At baseline, 441 subjects were coded positive for CVD (prevalent cases). Incident cases numbered 126. Multiple logistic regression analyses showed that, along with sex, age, smoking, HDL/total cholesterol ratio, and hypertension, HOMA-IR was an independent predictor of both prevalent and incident CVD. A 1-unit increase in (log)HOMA-IR value was associated with an odds ratio for prevalent CVD at baseline of 1.31 (95% CI 1.10-1.56, P = 0.002) and for incident CVD during follow-up of 1.56 (95% CI 1.14-2.12, P < 0.001). CONCLUSIONS: HOMA-IR is an independent predictor of CVD in type 2 diabetes. The improvement of insulin resistance might have beneficial effects not only on glucose control but also on CVD in patients with type 2 diabetes. PMID- 12087011 TI - A prospective study of obesity and risk of coronary heart disease among diabetic women. AB - OBJECTIVE: To examine the relationship of obesity, measured as BMI, and weight change to incidence of coronary heart disease (CHD) among women with diabetes. RESEARCH DESIGN AND METHODS: We followed 5,897 women with type 2 diabetes in the Nurses' Health Study for < or = 20 years. Women were aged 40-74 years and had no history of cardiovascular disease or cancer at the beginning of the follow-up period. BMI values from three time points (age 18 years, year 1976, and current) were derived from the reported height (1976) and corresponding reported weight. Weight changes between age 18 years and 1976 and after diagnosis of diabetes were calculated. Women reported diagnoses of diabetes and CHD every 2 years. Incident CHD cases were confirmed by medical record review. RESULTS: During follow-up, we documented 418 incident cases of CHD (236 of nonfatal myocardial infarction and 182 of fatal CHD). After adjustment for age, smoking, and other coronary risk factors, current BMI was strongly associated with increased risk of CHD among diabetic women. The multivariate relative risks across increasing categories of BMI (<23.0, 23.0-24.9, 25.0-26.9, 27.0-29.9, 30.0-34.9, and > or = 35.0 kg/m(2)) were 1.0, 1.58, 1.85, 1.95, 2.80, and 3.21, respectively (P for trend <0.001). Increasing BMI values from age 18 years to 1976, before diagnosis of diabetes, were also positively associated with risk of CHD. Weight gain before the diagnosis of diabetes was related to increased risk of CHD. In contrast, weight change after diagnosis of diabetes was not associated with risk of CHD. CONCLUSIONS: These findings provide strong evidence that obesity and weight gain before diagnosis of diabetes are associated with future risk of CHD among women with type 2 diabetes. PMID- 12087012 TI - Diabetes care in the U.S. and Canada. AB - OBJECTIVE: To compare the glycemic control of patients with type 1 diabetes treated in the U.S. and Canada. RESEARCH DESIGN AND METHODS: A large multicenter randomized clinical trial conducted in the U.S. and Canada was analyzed. Patients with type 1 diabetes, screened from 1983 to 1989 for enrollment in the Diabetes Control and Complications Trial (DCCT), were categorized as treated in the U.S. (n = 2,604) or Canada (n = 245). HbA(1c) levels were compared between U.S. and Canadian patients, both before and after adjustment for predictors of HbA(1c). RESULTS: In general, volunteers screened for the DCCT were highly educated and following healthy lifestyles. Canadians were somewhat younger (25 vs. 27 years of age, P = 0.002), less likely to be college educated (62 vs. 71%, P = 0.002), more likely to receive care through a family doctor (41 vs. 28%, P = 0.001), and had a higher frequency of out-patient visits (4 vs. 3 per year, P = 0.004). Despite these differences in health care delivery, the mean HbA(1c) at baseline was identical in the two countries (8.9 vs. 9.0, P = 0.40). Adjustment for demographic, lifestyle, and clinical predictors of HbA(1c) yielded similar findings (9.0 vs. 9.2, P = 0.15). Equal percentages of American and Canadian patients who were screened ultimately entered the trial (21 vs. 19%, P = 0.20), and those randomized to conventional care achieved similar mean HbA(1c) levels (9.1 vs. 9.2, P = 0.50). CONCLUSIONS: Differences in care delivery patterns do not yield large differences in glycemic control for patients with type 1 diabetes who were recruited in the U.S. and Canada for a large randomized trial. PMID- 12087013 TI - A prospective analysis of marital relationship factors and quality of life in diabetes. AB - OBJECTIVE: To prospectively assess the relation between marital relationship domains (i.e., intimacy and adjustment) and health-related quality of life (HRQoL) and glycemic control of individuals with diabetes. RESEARCH DESIGN AND METHODS: A previous cross-sectional study assessed the relationship between marital quality and adaptation to diabetes for 78 insulin-treated adults. Each participant completed two marital quality measures (Spanier Dyadic Adjustment Scale and Personal Assessment of Intimacy in Relationships Scale) at time 1 (T1). A total of 61 subjects were contacted two years later (time 2 [T2]) and re administered four HRQoL measures (Diabetes Quality of Life Scale, Medical Outcomes Study Health Survey-SF36, Problem Areas in Diabetes Scale, and Positive and Negative Affect Schedule). Glycemic control was assessed by HbA(1c). Demographic data (age, sex, type and duration of diabetes, years married, other medical problems, family history, and years of education) were gathered from charts and questionnaires. Linear regression analyses were used to assess the ability of the two marital measures at T1 to predict HRQoL at T2, controlling for confounding HRQoL and baseline subject variables. RESULTS: Both of the marital quality measures at T1 were prospective predictors of aspects of HRQoL at T2. Less diabetes-related distress at T2 was predicted by better marital adjustment at T1. Greater satisfaction with aspects of the diabetes care regimen at T2 was predicted by better marital adjustment and greater perceived marital intimacy at T1. Neither marital measure predicted general HRQoL or glycemic control. CONCLUSIONS: For insulin-treated adults with diabetes, quality of marriage prospectively predicts diabetes-related quality of life. This study highlights the sensitivity of diabetes-specific measures. Future work should explore the value of interventions that target the spouse and/or couple. PMID- 12087014 TI - Self-management education for adults with type 2 diabetes: a meta-analysis of the effect on glycemic control. AB - OBJECTIVE: To evaluate the efficacy of self-management education on GHb in adults with type 2 diabetes. RESEARCH DESIGN AND METHODS: We searched for English language trials in Medline (1980-1999), Cinahl (1982-1999), and the Educational Resources Information Center database (ERIC) (1980-1999), and we manually searched review articles, journals with highest topic relevance, and reference lists of included articles. Studies were included if they were randomized controlled trials that were published in the English language, tested the effect of self-management education on adults with type 2 diabetes, and reported extractable data on the effect of treatment on GHb. A total of 31 studies of 463 initially identified articles met selection criteria. We computed net change in GHb, stratified by follow-up interval, tested for trial heterogeneity, and calculated pooled effects sizes using random effects models. We examined the effect of baseline GHb, follow-up interval, and intervention characteristics on GHb. RESULTS: On average, the intervention decreased GHb by 0.76% (95% CI 0.34 1.18) more than the control group at immediate follow-up; by 0.26% (0.21% increase - 0.73% decrease) at 1-3 months of follow-up; and by 0.26% (0.05-0.48) at > or = 4 months of follow-up. GHb decreased more with additional contact time between participant and educator; a decrease of 1% was noted for every additional 23.6 h (13.3-105.4) of contact. CONCLUSIONS: Self-management education improves GHb levels at immediate follow-up, and increased contact time increases the effect. The benefit declines 1-3 months after the intervention ceases, however, suggesting that learned behaviors change over time. Further research is needed to develop interventions effective in maintaining long-term glycemic control. PMID- 12087015 TI - C-reactive protein, diabetes, and attendance at religious services. AB - OBJECTIVE: Previous studies have shown an association between attendance at religious services and health, particularly cardiovascular morbidity and mortality. People with diabetes have higher risk of cardiovascular mortality and higher C-reactive protein (CRP) levels than people without diabetes. The purpose of this study was to explore the relationship between religious attendance and CRP in people with diabetes. RESEARCH DESIGN AND METHODS: This study used cross sectional survey and examination of a nationally representative sample of noninstitutionalized U.S. adults aged > or = 40 years, derived from the National Health and Nutrition Examination Survey III 1988-1994 (n = 10,059). There were 556 people with diabetes. The primary outcome measure was the presence of elevated CRP. RESULTS: There were no differences between people with diabetes compared with people without diabetes in the percentage of those who attended religious services (62.29 vs. 62.0%, P = 0.36). Religious service nonattenders with diabetes were more likely than attenders to have an elevated CRP (odds ratio [OR] 2.17 [95% CI 1.15-4.09]). In people without diabetes, the association between attendance and CRP was not significant (P > 0.05). Among people with diabetes, after adjusting for demographic variables, health status, smoking, social support, mobility, and BMI, the association between religious attendance and CRP remained significant for respondents with diabetes (1.90 [1.03-3.51]). CONCLUSIONS: These findings revealed that people with diabetes who have not attended religious services in the previous year are more likely to have elevated levels of CRP. Further research should be conducted to evaluate this association to improve our understanding of the psychological and religious factors that influence diabetes. PMID- 12087016 TI - Homeostasis model assessment of insulin resistance in relation to the incidence of cardiovascular disease: the San Antonio Heart Study. AB - OBJECTIVE: The prospective association between insulin levels and risk of cardiovascular disease (CVD) is controversial. The objective of the present study was to investigate the relationship of the homeostasis model assessment of insulin resistance (HOMA-IR), as well as insulin levels, with risk of nonfatal and fatal CVD over the 8-year follow-up of the San Antonio Heart Study. RESEARCH DESIGN AND METHODS: Between 1984 and 1988, randomly selected Mexican-American and non-Hispanic white residents of San Antonio participated in baseline examinations that included fasting blood samples for glucose, insulin, and lipids, a glucose tolerance test, anthropometric measurements, and a lifestyle questionnaire. Between 1991 and 1996, 2,569 subjects who were free of diabetes at baseline were reexamined using the same protocol. RESULTS: Over the follow-up period, 187 subjects experienced an incident cardiovascular event (heart attack, stroke, heart surgery, angina, or CVD death). Logistic regression analysis indicated that risk of a CVD event increased across quintiles of HOMA-IR after adjustment for age, sex, and ethnicity (P for trend <0.0001; quintile 5 vs. quintile 1, odds ratio [OR] 2.52, 95% CI 1.46-4.36). Additional adjustment for LDL, triglyceride, HDL, systolic blood pressure, smoking, alcohol consumption, exercise, and waist circumference only modestly reduced the magnitude of these associations (P for trend 0.02; quintile 5 vs. quintile 1, OR 1.94, 95% CI 1.05-3.59). Furthermore, there were no significant interactions between HOMA-IR and ethnicity, sex, hypertension, dyslipidemia, glucose tolerance (impaired glucose tolerance versus normal glucose tolerance), or obesity. The magnitude and direction of the relationship between insulin concentration and incident CVD were similar. CONCLUSIONS: We found a significant association between HOMA-IR and risk of CVD after adjustment for multiple covariates. The topic remains controversial, however, and additional studies are required, particularly among women and minority populations. PMID- 12087017 TI - Reproducibility of glucose measurements using the glucose sensor. AB - OBJECTIVE: Recent studies have confirmed that improved glycemic control decreases the risk of diabetic complications in type 1 and type 2 diabetic patients. The Minimed glucose sensor allows continuous 72-h glucose monitoring and represents a potentially important tool to improve diabetes management. Its use is currently limited to the health care team. Our aim was to evaluate the reproducibility of data provided by the device by comparing data provided by two sensors worn simultaneously by the same subject. RESEARCH DESIGN AND METHODS: A total of 11 subjects (6 type 1 and 3 type 2 diabetic patients and 2 healthy subjects) agreed to wear two sensors and perform at least four daily finger-stick glucose determinations during 72 h. The simultaneous glucose values provided by the sensors were compared. To determine the clinical implications of the glucose data, each day was divided into eight periods, and for each period the glucose range was rated as satisfactory, too high, or too low by a blinded clinician experienced in interpreting glucose sensor data in the clinical setting. The evaluation of glycemic levels based on the recordings of the two sensors were compared for each paired time interval. RESULTS: We discarded 18% of the sensor data for technical reasons. Examined as a group, the remaining 3,370 paired data points in all 11 patients were highly correlated (r = 0.84). However, when individual pairs were evaluated, large differences in the glucose values were apparent, with differences of >10% in 70% of the measurements and >50% in 7% of the measurements. Moreover, clinical evaluation of the glucose range provided simultaneously by two sensors was concordant for only 65% of the evaluation periods. CONCLUSION: In a real-life setting, the accuracy of data provided by the Minimed glucose sensor may be less than expected. To avoid therapeutic errors, sensor findings should be confirmed by independent means before clinical decisions are made. PMID- 12087018 TI - IA-2 antibody-negative status predicts remission and recovery of C-peptide levels in type 1 diabetic patients treated with cyclosporin. AB - OBJECTIVE: The use of cyclosporin in recent-onset type 1 diabetes has demonstrated the potential for immune intervention in the treatment and prevention of the disease. However, a proportion of patients failed to respond to cyclosporin treatment. Indicators of resistance to immune intervention would be valuable for the most effective use of such therapies in disease prevention. The aim of this study was to determine whether presence of IA-2 antibodies is such a marker. RESEARCH DESIGN AND METHODS: IA-2 antibodies were determined by radioligand binding assay in sera from patients recruited into the Canadian European cyclosporin trial. Insulin dose requirements and glucagon-stimulated C peptide secretion were analyzed in patients grouped according to IA-2 antibody status at entry. RESULTS: Cyclosporin treatment had no significant effect on frequency of IA-2 antibodies during the 1 year of treatment. Cyclosporin caused significant reduction in insulin requirements and significant increases in C peptide secretion mainly in patients negative for IA-2 antibodies. Analysis of GAD antibodies in combination with antibodies to IA-2 indicated that the group most resistant to cyclosporin were IA-2 antibody positive, GAD antibody negative. CONCLUSIONS: The results demonstrate that IA-2 antibody analysis is valuable in identifying individuals for whom immunosuppressive treatment would be most effective. PMID- 12087019 TI - Atorvastatin and micronized fenofibrate alone and in combination in type 2 diabetes with combined hyperlipidemia. AB - OBJECTIVE: This study evaluated the effect of a atorvastatin-fenofibrate combination on lipid profile, in comparison to each drug alone, in patients with type 2 diabetes and combined hyperlipidemia (CHL). RESEARCH DESIGN AND METHODS: A total of 120 consecutive patients, who were free of coronary artery disease (CAD) at entry, were studied for a period of 24 weeks. These patients were randomly assigned to atorvastatin (20 mg/day, n = 40), micronized fenofibrate (200 mg/day, n = 40), or a combination of both (atorvastatin 20 mg/day plus fenofibrate 200 mg/day, n = 40). The effect of treatment on LDL cholesterol, triglycerides (TGs), HDL cholesterol, apolipoprotein A-I and B, lipoprotein(a), and plasma fibrinogen (PF) was recorded. Moreover, the percentage of patients that reached the American Diabetes Association treatment goals and the estimated CAD risk status were calculated. RESULTS: No patient was withdrawn from the study because of side effects. The atorvastatin-fenofibrate combination reduced total cholesterol by 37%, LDL cholesterol by 46%, TGs by 50%, and PF by 20%, whereas it increased HDL cholesterol by 22% (P < 0.0001 for all). These changes were significantly better than those of both monotherapies. Of the patients on drug combination, 97.5% reached the LDL cholesterol treatment goal of <100 mg/dl, 100% reached the desirable TG levels of <200 mg/dl, and 60% reached the optimal HDL cholesterol levels of >45 mg/dl. These rates were significantly higher than those of both monotherapies. Combined treatment reduced the 10-year probability for myocardial infarction from 21.6 to 4.2%. CONCLUSIONS: The atorvastatin-fenofibrate combination has a highly beneficial effect on all lipid parameters and PF in patients with type 2 diabetes and CHL. It improved patients' CAD risk status significantly more than each drug alone. PMID- 12087020 TI - Development of a prediction equation for insulin sensitivity from anthropometry and fasting insulin in prepubertal and early pubertal children. AB - OBJECTIVE: To test the utility of homeostasis model assessment (HOMA) in predicting insulin sensitivity [x10(- 4) min(-1)/(microIU/ml)] in children and to develop and compare two new prediction equations for insulin sensitivity in children using demographic and anthropometric measures in the presence or absence of fasting insulin. RESEARCH DESIGN AND METHODS: We studied 156 white and African American children with complete data (mean age 9.7 +/- 1.8 years, 87.8% Tanner Stage 1 or 2). For development of new equations, two-thirds of the children were randomly assigned to a development group, whereas the remaining children were assigned to a cross-validation group. RESULTS: A modified HOMA equation accurately predicted insulin sensitivity, but its utility is similar to fasting insulin alone. Demographic and anthropometric measures alone did not predict insulin sensitivity accurately, even when precise measures of body composition were included in the prediction model. Ethnicity, calf skinfold, and fasting insulin together explained 73% of the variance in insulin sensitivity and accurately predicted insulin sensitivity. The regression of measured versus predicted insulin sensitivity in the cross-validation group was not significantly different from the line of identity (P > 0.05). Mean difference between measured and predicted insulin sensitivity was also not significant (P > 0.05). Some bias was apparent, particularly in white boys. CONCLUSIONS: Ethnicity, calf skinfold, and fasting insulin can accurately predict insulin sensitivity with greater precision than HOMA or fasting insulin alone (R(2) = 0.73). Future studies, however, are needed to examine whether a universal equation is possible. A cross validated prediction equation may be useful in population-based studies when complex measures of insulin sensitivity are not available. PMID- 12087021 TI - Aggressive lipid lowering does not improve endothelial function in type 2 diabetes: the Diabetes Atorvastatin Lipid Intervention (DALI) Study: a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: Endothelial dysfunction is considered an important early marker of atherosclerosis and cardiovascular risk and is currently used as a surrogate end point for cardiovascular risk in clinical trials. Type 2 diabetic patients show a characteristic dyslipidemia. Aggressive lipid lowering might be an effective method to improve endothelial function in these patients. RESEARCH DESIGN AND METHODS: A randomized, double-blind, placebo-controlled trial was completed to study the effect of 30 weeks' administration of atorvastatin 10 mg and 80 mg on endothelial function, as assessed by B-mode ultrasound of the brachial artery, in 133 patients with type 2 diabetes without a history of cardiovascular disease. RESULTS: Patients with diabetes and diabetic dyslipidemia had considerable endothelium-dependent and endothelium-independent dysfunction; mean flow-mediated vasodilation (SD) was 3.16% (3.56), and mean response on sublingual nitroglycerin was 6.58% (6.04). Despite substantial lowering of all atherogenic lipid parameters, no improvement of endothelium-dependent vasodilatation was found (P > 0.8). CONCLUSIONS: We observed considerable baseline endothelium-dependent and endothelium-independent dysfunction in patients with diabetes and diabetic dyslipidemia without a history of cardiovascular disease. Aggressive lipid lowering by administration of atorvastatin, resulting in substantial improvement of the lipid profile, did not reverse endothelial dysfunction. PMID- 12087022 TI - Insulin sensitivity and beta-cell function in women with polycystic ovary syndrome. AB - OBJECTIVE: To evaluate insulin sensitivity (IS) and beta-cell function (beta F) in lean and obese women with polycystic ovary syndrome (PCOS), either separately or by using a disposition index (DI). RESEARCH DESIGN AND METHODS: A total of 64 women with PCOS and 20 healthy women were examined by anthropometry, oral glucose tolerance tests (OGTTs), and insulin tolerance tests. Statistical analysis used one-way ANOVA, Kruskal-Wallis, and Mann-Whitney U tests, as appropriate. RESULTS: A significantly higher waist-to-hip ratio (P < 0.0001) was found in both lean and obese women with PCOS. Higher basal blood glucose (P < 0.004) and blood glucose values at 3 h of OGTT (P < 0.008) were found in lean and obese PCOS subjects in comparison with control subjects. Insulin resistance by homeostasis model assessment (P < 0.007) was significantly higher in obese PCOS than in control or lean PCOS subjects. Early-phase insulin secretion (insulinogenic index [Delta I/Delta G(30-0), where I is insulin and G is glucose]; P < 0.0007) was significantly higher in both lean and obese PCOS subjects than in healthy women. All tested combinations of parameters of IS and beta F (DIs) followed a physiological hyperbolic relationship. Significantly lower values of the fasting state-derived DIs were found (all P < 0.05) in obese PCOS subjects. Significantly higher values of all of these indexes derived from nonfasting values were found in lean PCOS as compared with control and obese PCOS subjects (all P < 10(-3)). CONCLUSIONS: Increased beta F was found even in lean individuals with PCOS. Insulin hypersecretion is thus probably connected to the pathogenesis of PCOS. PMID- 12087023 TI - Relation of drinking alcohol to atherosclerotic risk in type 2 diabetes. AB - OBJECTIVE: The effects of drinking alcohol on atherosclerotic risks were investigated in 194 type 2 diabetic patients to determine whether drinking alcohol influences risk of atherosclerosis in diabetic subjects. RESEARCH DESIGN AND METHODS: The subjects were divided by the degree of their average weekly alcohol consumption into three groups: nondrinkers, light drinkers (ethanol consumption <210 g/week), and heavy drinkers (ethanol consumption > or = 210 g/week). The degree of atherosclerotic progression was evaluated using aortic pulse wave velocity (a-PWV), and possible atherosclerotic risks were evaluated using known atherosclerotic risk factors. RESULTS: a-PWV was significantly lower in light drinkers than in nondrinkers and heavy drinkers, but there was no significant difference in a-PWV between nondrinkers and heavy drinkers. Systolic blood pressure, HDL cholesterol, and triglyceride levels were significantly higher in heavy drinkers than in nondrinkers and light drinkers, whereas there was no significant difference in these levels between nondrinkers and light drinkers. The mean levels of BMI and blood HbA(1c), uric acid, and fibrinogen were not different between the three groups. There were significant positive correlations of a-PWV with age and systolic blood pressure and weak but significant negative correlations of a-PWV with alcohol consumption and HDL cholesterol level. CONCLUSIONS: Light drinking, but not heavy drinking, has preventive effects on atherosclerosis in type 2 diabetic subjects. The known beneficial effects of drinking alcohol on blood lipids and fibrinogen may not be involved in the preventive effect of light drinking on atherosclerosis in diabetic subjects. PMID- 12087024 TI - European Association for the Study of Diabetes (EASD) 2001 meeting. PMID- 12087025 TI - New strategies for the treatment of diabetic dyslipidemia. PMID- 12087026 TI - Manipulating the type 1 diabetes disease process, man versus mouse. PMID- 12087027 TI - Endothelial vasodilation effects of statins in type 2 diabetic patients: response to van Venrooij et al. PMID- 12087028 TI - The cost of self-monitoring of blood glucose is an important factor limiting glycemic control in diabetic patients. PMID- 12087029 TI - Association between vitamin D receptor genotype and age of onset in juvenile Japanese patients with type 1 diabetes. PMID- 12087030 TI - Association between plasma thrombin-activatable fibrinolysis inhibitor levels and activated protein C in normotensive type 2 diabetic patients. PMID- 12087031 TI - Metabolic syndrome in American Indians. PMID- 12087032 TI - Fasting hyperglycemia predicts the magnitude of postprandial hyperglycemia: implications for diabetes therapy. PMID- 12087033 TI - Dysfunction of active transport of blood-retinal barrier in patients with clinically significant macular edema in type 2 diabetes. PMID- 12087034 TI - Necrobiosis lipodica is a clinical feature of maturity-onset diabetes of the young. PMID- 12087035 TI - Atorvastatin, diabetic dyslipidemia, and cognitive functioning. PMID- 12087036 TI - Rosiglitazone in combination with glimepiride plus metformin in type 2 diabetic patients. PMID- 12087037 TI - Preobesity in World Health Organization classification involves the metabolic syndrome in Japanese. PMID- 12087038 TI - Prevalence of metabolic syndrome among HIV patients. PMID- 12087039 TI - Effect of losartan on plasma C-reactive protein in type 2 diabetic patients with microabluminuria. PMID- 12087040 TI - The value of self-monitoring of blood glucose. PMID- 12087041 TI - Evidence-based nutritional recommendations for the treatment and prevention of diabetes and related complications: a European perspective. PMID- 12087044 TI - Insulin resistance after renal transplantation. PMID- 12087047 TI - A randomized controlled trial using glycemic plus fetal ultrasound parameters versus glycemic parameters to determine insulin therapy in gestational diabetes with fasting hyperglycemia. PMID- 12087049 TI - New dietary guidelines from the American Diabetes Association. PMID- 12087050 TI - American Diabetes Association evidence-based nutrition principles and recommendations are not based on evidence. PMID- 12087052 TI - Science at a crossroads. AB - Science is entering an alliance with the economy that will speed the effect of innovation through society. Despite the slowdown of the 'new economy', a cascade paradigm of innovation appears key to increasing the rate of economic growth. Yet for science to continue to thrive and make this contribution to innovation, it must traverse at least three key crossroads. First, while life sciences have built a strong advocacy model to secure growing federal research funding, the physical sciences (including mathematics and engineering) have not and must now do so to thrive. Second, the drop in the numbers of physical scientists and engineers must be reversed if we are to have the talent to maintain a strong trend of scientific advance. Third, although science advances are increasingly interdisciplinary and occurring in the space between the historic science stovepipes, the organization of federal science support is largely unchanged since the beginning of the cold war. While a decentralized model has value, we must also consider new approaches that encourage deeper cooperation across science sectors and agencies. PMID- 12087053 TI - The role of tumor lymphangiogenesis in metastatic spread. AB - The high mortality rates associated with cancer can be attributed to the metastatic spread of tumor cells from the site of their origin. Tumor cells invade either the blood or lymphatic vessels to access the general circulation and then establish themselves in other tissues. Clinicopathological data suggest that the lymphatics are an initial route for the spread of solid tumors. Detection of sentinel lymph nodes by biopsy provides significant information for staging and designing therapeutic regimens. The role of angiogenesis in facilitating the growth of solid tumors has been well established, but the presence of lymphatic vessels and the relevance of lymphangiogenesis to tumor spread are less clear. Recently, the molecular pathway that signals for lymphangiogenesis and relatively specific markers for lymphatic endothelium have been described allowing analyses of tumor lymphangiogenesis to be performed in animal models. These studies demonstrate that tumor lymphangiogenesis is a major component of the metastatic process and implicate members of the VEGF family of growth factors as key mediators of lymphangiogenesis in both normal biology and tumors. PMID- 12087054 TI - Hepatocyte telomere shortening and senescence are general markers of human liver cirrhosis. AB - Telomere shortening limits the number of cell divisions of primary human cells and might affect the regenerative capacity of organ systems during aging and chronic disease. To test whether the telomere hypothesis applies to human cirrhosis, the telomere length was monitored in cirrhosis induced by a broad variety of different etiologies. Telomeres were significantly shorter in cirrhosis compared with noncirrhotic samples independent of the primary etiology and independent of the age of the patients. Quantitative fluorescence in situ hybridization showed that telomere shortening was restricted to hepatocytes whereas lymphocytes and stellate cells in areas of fibrosis had significantly longer telomere reserves. Hepatocyte-specific telomere shortening correlated with senescence-associated beta-galactosidase staining in 84% of the cirrhosis samples, specifically in hepatocytes, but not in stellate cells or lymphocytes. Hepatocyte telomere shortening and senescence correlated with progression of fibrosis in cirrhosis samples. This study demonstrates for the first time that cell type-specific telomere shortening and senescence are linked to progression of human cirrhosis. These findings give a novel explanation for the pathophysiology of cirrhosis, indicating that fibrotic scarring at the cirrhosis stage is a consequence of hepatocyte telomere shortening and senescence. The data imply that future therapies aiming to restore regenerative capacity during aging and chronic diseases will have to ensure efficient targeting of specific cell types within the affected organs. PMID- 12087055 TI - Characterization of aquaporin-4 in muscle and muscular dystrophy. AB - Aquaporins are a growing family of transmembrane proteins that transport water and, in some cases, glycerol and urea across cellular membranes. Aquaporin-4 (AQP4) is enriched at the sarcolemma of skeletal muscle and may play a role in accommodating the rapid changes in cell volume and hydrostatic forces that occur during contraction in order to prevent damage to the sarcolemma. Recent evidence has shown that AQP4 is absent in dystrophin-deficient mdx mice, suggesting that AQP4 associates with dystrophin and has a role in the dystrophic process. To examine the relationship between aquaporins and muscle disease, and between aquaporins and dystrophin, we have investigated aquaporin expression in various mouse models of muscular dystrophy and cardiomyopathy before and after the onset of pathology. We find that AQP4 is expressed in prenecrotic mdx muscle despite the absence of dystrophin and that AQP4 is lost after the onset of muscle degeneration. Analysis of various dystrophin transgenic mice reveals that AQP4 is lost even when the dystrophin-glycoprotein complex is present, suggesting that loss of AQP4 is not directly resulting from loss of the DGC. AQP4 was also lost in muscular dystrophies caused by primary mutations in the sarcoglycan genes. Taken together, our data demonstrate that AQP4 loss in skeletal muscle correlates with muscular dystrophy and is a common feature of pathogenesis. PMID- 12087056 TI - Hepatocyte growth factor enhances endothelial cell barrier function and cortical cytoskeletal rearrangement: potential role of glycogen synthase kinase-3beta. AB - The stabilization of endothelial cell (EC) barrier function within newly formed capillaries is a critical feature of angiogenesis. We examined human lung EC barrier regulation elicited by hepatocyte growth factor (HGF), a recognized angiogenic factor and EC chemoattractant. HGF rapidly and dose-dependently elevated transendothelial electrical resistance (TER) of EC monolayers (>50% increase at 100 ng/ml), with immunofluorescence microscopic evidence of both cytoplasmic actin stress fiber dissolution and strong augmentation of the cortical actin ring. HGF rapidly stimulated phosphatidylinositol 3'-kinase, ERK, p38 mitogen-activated protein kinase, and protein kinase C activities. Pharmacological inhibitor studies demonstrated each pathway to be intimately involved in HGF-induced increases in TER, cortical actin thickening, and phosphorylation of the Ser/Thr glycogen synthase kinase-3beta (GSK-3beta), a potential target for the HGF barrier-promoting response. GSK-3beta phosphorylation was strongly correlated with reductions in both HGF-induced TER and enhanced beta-catenin immunoreactivity observed at cell-cell junctions. Our data suggest a model in which HGF-mediated EC cytoskeletal rearrangement and barrier enhancement depend critically on the activation of a complex kinase cascade that converges at GSK-3beta to increase the availability of beta-catenin, thereby enhancing endothelial junctional integrity and vascular barrier function. PMID- 12087057 TI - Accelerated wound healing in tumor necrosis factor receptor p55-deficient mice with reduced leukocyte infiltration. AB - To clarify biological roles of tumor necrosis factor receptor p55 (TNF-Rp55) mediated signals in wound healing, skin excisions were prepared in BALB/c (WT) and TNF-Rp55-deficient (KO) mice. In WT mice, the wound area was reduced to 50% of the original area 6 days after injury, with angiogenesis and collagen accumulation. Histopathologically, reepithelialization rate was approximately 80% 6 days. Myeloperoxidase activity and macrophage recruitment were the most evident 1 and 6 days after injury, respectively. Gene expression of adhesion molecules, interleukin 1alpha (IL-1alpha), IL-1beta, monocyte chemoattractant protein 1, macrophage inflammatory protein 1alpha (MIP-1alpha), MIP-2, transforming growth factor beta1 (TGF-beta1) connective tissue growth factor (CTGF), vascular endothelial growth factor (VEGF), Flt-1, and Flk-1 was enhanced at the wound site. In KO mice, an enhancement in angiogenesis, collagen content, and reepithelialization was accelerated with the increased gene expression of TGF beta1, CTGF, VEGF, Flt-1, and Flk-1 at the wound sites, resulting in accelerated wound healing compared with WT mice. In contrast, leukocyte infiltration, mRNA expression of adhesion molecules, and cytokines were significantly reduced in KO mice. These observations suggest that TNF-Rp55-mediated signals have some role in promoting leukocyte infiltration at the wound site and negatively affect wound healing, probably by reducing angiogenesis and collagen accumulation. PMID- 12087058 TI - Heterotopic implantation alters the regulation of apoptosis and the cell cycle and generates a new metastatic site in a human pancreatic tumor xenograft model. AB - Differences in growth and in response to antineoplastic drugs between s.c. and orthotopically implanted tumors in nude mice and between the primary tumor and the metastases in human tumors suggest that implantation site may alter the molecular regulation of tumor cells. We assessed the influence of implantation site on cell cycle and apoptotic regulation and the possible contribution of the implantation site in directing the choice of metastatic site by comparing the behavior of tumor aliquots of two human pancreatic xenografts (NP18 and NP9) implanted in the organ where the tumor grows (orthotopically), in heterotopic sites (the site of metastases (liver), and in nonmetastatic sites (subcutis and colon). We observed that implantation site changes tumor growth by altering apoptotic or cell cycle regulation in a tumor-specific manner. In the NP18 tumor it occurs by altering apoptotic induction and activation of the Bad/Bcl XL/caspase-3 pathway through AKT and Erk regulation, but in the NP9 tumor by changing the activation and/or expression of the proteins that regulate the cell cycle (Erk, PCNA, and cyclin B1). We also observed that implantation site alters the metastatic pattern of the NP9 tumor, originating a new metastatic site. PMID- 12087059 TI - N-glycans of sphingosine 1-phosphate receptor Edg-1 regulate ligand-induced receptor internalization. AB - Endothelial differentiation gene-1 product (Edg-1) is a G-protein-coupled receptor (GPCR) for the platelet derived bioactive lipid mediator sphingosine 1 phosphate (Sph-1-P). Recent studies have shown that in response to Sph-1-P, Edg-1 mediates various signaling pathways through downstream signaling molecules, such as MAP kinase and calcium, via heterotrimeric G-proteins. We found for the first time that Edg-1 is glycosylated in its amino-terminal extracellular portion, and further identified the specific glycosylation site as asparagine 30 by creating a nonglycosylated mutant of Edg-1 (N30D-Edg-1) and transfecting it into cell lines. The nonglycosylated mutant receptors, resembling their wild-type controls, were predominantly expressed in the plasma membrane. Although there was no difference in ligand binding ability and ligand-induced MAP kinase activation in the wild type and mutant receptors, nonglycosylated Edg-1 was much less responsive for ligand-induced internalization. Unlike the wild-type receptor, which was associated with the caveolae, nonglycosylated N30D-Edg-1 was dispersed broadly in the membrane fractions separated by sucrose density gradient centrifugation, suggesting that internalization and microdomain localization of N-glycosylated Edg-1 might be related. Although the precise molecular mechanism of the internalization of the N-glycosylated Edg-1 localized in the microdomain remains to be examined, the present study suggested that the presence of N-linked glycan in the receptor may play a regulatory role in the receptor dynamics in ligand stimulated mammalian cells. PMID- 12087060 TI - Anti-inflammatory cooperativity of corticosteroids and norepinephrine in rheumatoid arthritis synovial tissue in vivo and in vitro. AB - Corticosteroids (CS) and norepinephrine (NE) support each other's biological effects. Thus, deficiency of cortisol and reduced synovial sympathetic innervation (SSI) may be proinflammatory in rheumatoid arthritis (RA). This study tested the anti-inflammatory cooperativity of CS and NE in human RA synovial tissue. In an in vivo study, 32 patients with RA (with prior CS therapy/without SSI: n=7; without prior CS therapy/with SSI: 6; with prior CS therapy/with SSI: 19) were investigated for synovial inflammation. In an in vitro study with synoviocytes from RA and OA patients, the separate and combined effects of cortisol and NE were studied. In the in vivo study, patients with prior CS therapy/with SSI showed lower secretion of synovial IL-8 than the other groups, lower synovial density of T cells and macrophages, and lower overall inflammation. In the in vitro study, a cooperative suppressive effect of NE (10( 6) M to 10(-8) M) and cortisol (10(-6) M and 10(-7) M) on secretion of IL-8 and TNF from primary early culture mixed RA synoviocytes was observed. This cooperative effect was not observed in OA synoviocytes. In the same RA and OA patients, the cooperative effect was lost in 3rd passage synovial fibroblasts. This study demonstrates the cooperativity of cortisol and NE for inhibition of proinflammatory mediators produced in the synovial tissue of RA patients. These results underscore that coupling of an efficient secretion of systemic cortisol together with local production of NE is important in order to lower synovial inflammation. PMID- 12087061 TI - Embryonic prostaglandin H synthase-2 (PHS-2) expression and benzo[a]pyrene teratogenicity in PHS-2 knockout mice. AB - The developmental role of prostaglandin H synthase-2 (PHS-2), which converts xenobiotics such as benzo[a]pyrene (B[a]P) to toxic free radical intermediates, is poorly understood. In this study, we determined the embryonic expression and teratological relevance of PHS-2 in pregnant CD-1 and B6/129S7 PHS-2 knockout mice. Wild-type (+/+) B6/129S7 dams given B[a]P on gestational day (GD) 10 had three times more fetal malformations than did +/- PHS-2-deficient dams (P<0.05). GD 10-13 CD-1 embryos had high PHS-2 protein expression, and both + /+ and +/- GD 19 B6/129S7 fetuses had more B[a]P-initiated malformations and postpartum lethality than did -/- littermates (P<0.05). Thus, embryonic PHS-2 is expressed constitutively during organogenesis and contributes substantially to B[a]P teratogenicity. PMID- 12087062 TI - Suppression of human tumor cell proliferation through mitochondrial targeting. AB - Intracellular calcium signaling plays a central role in cell proliferation. In leukemic cells, the calcium release-activated calcium channels provide a major pathway for calcium entry (I(CRAC)) perpetuating progression through the cell cycle. Although I(CRAC) is under mitochondrial regulation, targeting mitochondrial function has not been exploited to control malignant cell growth. The benzothiadiazine diazoxide, which depolarized respiration-dependent mitochondrial membrane potential, reduced the rate of proliferation and arrested human acute leukemic T cells in the G0/G1 phase. Diazoxide did not alter cellular energetics, but rather inhibited the mitochondria-controlled I(CRAC) and reduced calcium influx into tumor cells. The antiproliferative action of diazoxide was mimicked by removal of extracellular calcium or by the tyrphostin A9, an I(CRAC) inhibitor. Deletion of the mitochondrial genome, which encodes essential respiratory chain enzyme subunits, attenuated the inhibitory effect of diazoxide on I(CRAC)-mediated calcium influx and cell proliferation. Thus, manipulation of mitochondrial function and associated calcium signaling provides a basis for a novel anticancer strategy. PMID- 12087063 TI - Programming effects of short prenatal exposure to cortisol. AB - Recent studies have linked fetal exposure to a suboptimal intrauterine environment with adult hypertension. The aims of this study were twofold: 1) to see whether cortisol treatment administered to the ewe for 2 days at 27 days of gestation (term approximately 150 days) resulted in high blood pressure in offspring; 2) to study the effect of the same treatment on gene expression in the brain at 130 days of gestation and in lambs at 2 months of age. Mean arterial pressure was significantly higher in the adult female and male offspring of sheep treated with cortisol than in the control group (females: 89+/-2 mmHg vs. 81+/-2; P<0.05 and males: 102+/-4 mmHg vs. 91+/-3; P<0.05). Prenatal cortisol treatment led to up-regulation of angiotensinogen, AT1, MR, and GR mRNA in the hippocampus in fetuses at 130 days of gestation but not in the animals at 2 months of age. This is the first evidence that short prenatal exposure to cortisol programmed high blood pressure in the adult female and male offspring of sheep. Altered gene expression in the hippocampus could have a significant effect on the development of the hippocampus, and on postnatal behavior. PMID- 12087064 TI - Ligands of the peroxisome proliferator-activated receptors (PPAR-gamma and PPAR alpha) reduce myocardial infarct size. AB - This study was designed to investigate the effects of various chemically distinct activators of PPAR-gamma and PPAR-alpha in a rat model of acute myocardial infarction. Using Northern blot analysis and RT-PCR in samples of rat heart, we document the expression of the mRNA for PPAR-gamma (isoform 1 but not isoform 2) as well as PPAR-beta and PPAR-alpha in freshly isolated cardiac myocytes and cardiac fibroblasts and in the left and right ventricles of the heart. Using a rat model of regional myocardial ischemia and reperfusion (in vivo), we have discovered that various chemically distinct ligands of PPAR-gamma (including the TZDs rosiglitazone, ciglitazone, and pioglitazone, as well as the cyclopentanone prostaglandins 15D-PGJ2 and PGA1) cause a substantial reduction of myocardial infarct size in the rat. We demonstrate that two distinct ligands of PPAR-alpha (including clofibrate and WY 14643) also cause a substantial reduction of myocardial infarct size in the rat. The most pronounced reduction in infarct size was observed with the endogenous PPAR-gamma ligand, 15-deoxyDelta12,14 prostagalndin J2 (15D-PGJ2). The mechanisms of the cardioprotective effects of 15D-PGJ2 may include 1) activation of PPAR-alpha, 2) activation of PPAR-gamma, 3) expression of HO-1, and 4) inhibition of the activation of NF-kappaB in the ischemic-reperfused heart. Inhibition by 15D-PGJ2 of the activation of NF-kappaB in turn results in a reduction of the 1) expression of inducible nitric oxide synthase and the nitration of proteins by peroxynitrite, 2) formation of the chemokine MCP-1, and 3) expression of the adhesion molecule ICAM-1. We speculate that ligands of PPAR-gamma and PPAR-alpha may be useful in the therapy of conditions associated with ischemia-reperfusion of the heart and other organs. Our findings also imply that TZDs and fibrates may help protect the heart against ischemia-reperfusion injury. This beneficial effect of 15D-PGJ2 was associated with a reduction in the expression of the 1) adhesion molecules ICAM-1 and P selectin, 2) chemokine macrophage chemotactic protein 1, and 3) inducible isoform of nitric oxide synthase. 15D-PGJ2 reduced the nitration of proteins (immunohistological analysis of nitrotyrosine formation) caused by ischemia reperfusion, likely due to the generation of peroxynitrite. Not all of the effects of 15D-PGJ2, however, are due to the activation of PPAR-gamma. For instance, exposure of rat cardiac myocytes to 15D-PGJ2, but not to rosiglitazone, results in an up-regulation of the expression of the mRNA for heme-oxygenase-1 (HO-1). Taken together, these results provide convincing evidence that several, chemically distinct ligands of PPAR-gamma reduce the tissue necrosis associated with acute myocardial infarction. PMID- 12087065 TI - Adenoviral VEGF-C overexpression induces blood vessel enlargement, tortuosity, and leakiness but no sprouting angiogenesis in the skin or mucous membranes. AB - Vascular endothelial growth factors (VEGFs) and their receptors (VEGFRs) are important regulators of blood and lymphatic vessel growth and vascular permeability. The VEGF-C/VEGFR-3 signaling pathway is crucial for lymphangiogenesis, and heterozygous inactivating missense mutations of the VEGFR 3 gene are associated with hereditary lymphedema. However, VEGF-C can have potent effects on blood vessels because its receptor VEGFR-3 is expressed in certain blood vessels and because the fully processed form of VEGF-C also binds to the VEGFR-2 of blood vessels. To characterize the in vivo effects of VEGF-C on blood and lymphatic vessels, we have overexpressed VEGF-C via adenovirus- and adeno associated virus-mediated transfection in the skin and respiratory tract of athymic nude mice. This resulted in dose-dependent enlargement and tortuosity of veins, which, along with the collecting lymphatic vessels were found to express VEGFR-2. Expression of angiopoietin 1 blocked the increased leakiness of the blood vessels induced by VEGF-C whereas vessel enlargement and lymphangiogenesis were not affected. However, angiogenic sprouting of new blood vessels was not observed in response to AdVEGF-C or AAV-VEGF-C. These results show that virally produced VEGF-C induces blood vessel changes, including vascular leak, but its angiogenic potency is much reduced compared with VEGF in normal skin. PMID- 12087066 TI - Activation of oocytes by latrunculin A. AB - Actin depolymerization by latrunculin A (LAT-A) in mature starfish oocytes induces a massive calcium mobilization that results in the discharge of the cortical granules and in the elevation of the fertilization envelope. The Ca2+ liberation starts as a circumscribed subplasma membrane hotspot, which is followed by a flash of Ca2+ increase restricted to the cortical layer. Ca2+ propagates rapidly from these peripheral regions to the center of the oocyte, initiating calcium oscillations. Blockade of the inositol 1,4,5-trisphosphate receptors with heparin does not affect the liberation of Ca2+ at the initial hotspot or the cortical flash, but abolishes the centripetal spreading of the wave and the Ca2+ oscillations. In Ca2+-free medium, LAT-A also initiates Ca2+ release at a discrete cortical point, but then propagates throughout the cell without first forming the uniform cortical flash. The latter is thus linked to the influx of external Ca2+, somehow promoted by the depolymerization of cortical (microvillar) actin. The Ca2+ response to spermatozoa (i.e., peripheral hotspot, cortical flash, globalization of the signal) closely mimics that promoted by LAT A. Thus, the initial cortical release of Ca2+ promoted by the sperm may be due to the depolymerization of actin. PMID- 12087067 TI - Periadventitial fat releases a vascular relaxing factor. AB - Virtually all blood vessels are surrounded by adventitial fat. Adipocytes produce a host of vasoactive substances that may influence vascular contraction. We tested whether or not perivascular adipose tissue modulates contraction of aortic ring preparations. We studied aortic rings surrounded by periadventitial adipose tissue from adult Sprague-Dawley rats. At a maximum concentration of 300 nM angiotensin II, 6.5 microM serotonin, and 5 microM phenylephrine, the contractile response of intact rings was 95%, 80%, and 30% lower than that of vessels without periadventitial fat. The anticontractile effect of periadventitial fat was reduced by inhibition of ATP-dependent K+ channels with glibenclamide (3 microM) and by the tyrosine kinase inhibitor genistein (10 microM). Blocking NOS, cyclo oxygenase, cytochrome P450, or adenosine receptors did not restore the vascular response in intact vessels. The anticontractile effect of perivascular fat was present in Zucker fa/fa rats, suggesting that leptin receptors were not responsible. Transferring the bath solution from intact vessels, isolated periadventitial tissue, and cultured rat adipocytes to precontracted vessels lacking periadventitial fat resulted in a rapid relaxation. We suggest that perivascular adventitial adipose tissue releases a transferable adventitium derived relaxing factor that acts by tyrosine kinase-dependent activation of K+ channels in vascular smooth muscle cells. PMID- 12087068 TI - Critical involvement of p38 MAP kinase in pertussis toxin-induced cytoskeletal reorganization and lung permeability. AB - Bordetella pertussis is an important cause of infection in humans worldwide, with full expression of the syndrome associated with characteristic increases in lung permeability and airway edema. The exact cellular mechanisms by which pertussis toxin (PTX) exerts pulmonary toxicity remain unknown, but may involve its ability to ADP-ribosylate-specific G-proteins. We determined that PTX directly and reproducibly reduced lung endothelial and epithelial cell barrier function in vitro and in vivo assessed by decreases in transmonolayer electrical resistance (TER) and isolated perfused lung preparations. Alterations in lung permeability began approximately 30 min after PTX and were dependent on intrinsic ADP ribosyltransferase activity, as neither the cell binding beta-oligomer subunit or a genetically engineered PTX mutant (devoid of ADP-ribosyltransferase activity) altered TER. PTX-induced barrier dysfunction was associated with mild increases in F-actin stress fiber formation and causally linked to p38 MAP kinase activities. PTX-mediated p38 MAP kinase activation did not involve either p42/p44 ERK, p60src, Rho family of GTPases, or phosphatidylinositol-3' kinase pathways. PTX-mediated decreases in TER were temporally linked to phosphorylation of the actin binding proteins Hsp27 and caldesmon, known substrates for the Ser/Thr kinase MAPKAP2, whose activity is regulated by p38 MAP kinase. In addition to defining novel signaling pathways involved in PTX-induced respiratory pathophysiology, these data suggest that the direct cell-activating effects of PTX be carefully considered as a potential limitation to its use as a tool in signal transduction analysis. PMID- 12087069 TI - Redox-sensitive vascular smooth muscle cell proliferation is mediated by GKLF and Id3 in vitro and in vivo. AB - Reactive oxygen species such as superoxide and hydroxyl radicals have been implicated in the pathogenic growth of various cell types. The molecular mechanisms involved in redox-sensitive cell growth control are poorly understood. Stimulation of cultured vascular smooth muscle cells (VSMC) with xanthin/xanthin oxidase (X/XO) increases proliferation, whereas stimulation with hydrogen peroxide and Fe3+NTA (H-Fe) causes growth arrest of VSMC. Differential Display led to the identification of two novel, differentially regulated redox-sensitive genes. The dominant negative helix-loop-helix protein Id3 is induced by X/XO and down-regulated by H-Fe. The transcription factor gut-enriched Kruppel-like factor (GKLF) is induced by H-Fe but not by X/XO. Induction of GKLF and inhibition of Id3 via transfection experiments leads to growth arrest, whereas overexpression of Id3 and inhibition of GKLF cause cell growth. Id3 down-regulation is induced via binding of GKLF to the Id3 promotor and concomitantly reduced Id3 gene transcription rate. GKLF induction by H-Fe is mediated through hydroxyl radicals, p38MAP kinase-, calcium-, and protein synthesis-dependent pathways. Id3 is induced by X/XO via superoxide, calcium, p38, and p42/44 MAP kinase. GKLF induces and Id3 depresses expression of p21WAF1/Cip1, p27KIP1, p53. Induction of Id3 is accomplished by angiotensin II via superoxide release. A vascular injury mouse model revealed that Id3 is overexpressed in proliferating vascular tissue in vivo. These findings reveal novel mechanisms of redox-controlled cellular proliferation involving GKLF and Id3 that may have general implications for our understanding of vascular and nonvascular growth control. PMID- 12087070 TI - Androgen-dependent expression of FcgammaRIIB2 by thyrocytes from patients with autoimmune Graves' disease: a possible molecular clue for sex dependence of autoimmune disease. AB - Thyrocyte expression of HLA class I and class II antigens and related accessory molecules would convert these epithelial cells into functional antigen-presenting cells. Here we show that whereas normal thyrocytes express FcRn, Graves' disease thyrocytes also express FcgammaRIIB2. We further find that expression of FcgammaRIIB2, but not FcRn, is repressed by dihydrotestosterone. By mediating the uptake and transport of autoantibodies, we suggest that these IgG Fc receptors contribute in various ways to the onset and/or progression of autoimmune thyroid diseases. The androgen-mediated decrease of FcgammaRIIB2 expression in Graves' disease thyrocytes also provides a rationale for the predominant susceptibility of women to develop an autoimmune thyroid disease. Our findings open up a new prospect to autoimmunity, linking the role of the target organ to the sex dependence in autoimmune disease. PMID- 12087071 TI - The Duffy antigen/receptor for chemokines (DARC) and prostate cancer. A role as clear as black and white? AB - Prostate cancer is the most commonly diagnosed cancer and the second leading cause of cancer-related death among men in the United States. African American men have a 60% greater incidence of prostate cancer and a twofold higher mortality rate than Caucasian men. The Duffy antigen/receptor for chemokines (DARC) is a receptor expressed on erythrocytes and vascular endothelial cells that binds to and clears angiogenic chemokines. The DARC also functions as the erythrocyte receptor for invasion by malarial parasites. Approximately 70% of African Americans lack erythrocyte expression of the DARC as a genetic mechanism of protection against malaria infection. Given the importance of angiogenic chemokines in the development of tumor vascular networks and the chemokine binding properties of the DARC, the possibility that a lack of DARC expression on erythrocytes may represent an epigenetic factor that predisposes African American men to a greater incidence and mortality of prostate cancer should be considered. PMID- 12087072 TI - Protein tyrosine nitration and peroxynitrite. PMID- 12087073 TI - Steroids mediate the expression of cytoplasmic and membrane-linked components in human myometrial cells. AB - It is well known that the smooth muscle of the human myometrium is a target for the steroid hormones progesterone (P4) and estrogen. Progesterone is believed to participate in the maintenance of pregnancy, while estrogen is possibly involved in the process of parturition by promoting cervical dilatation. We examined the combined effects of P4 and 17beta-estradiol (E2) on components of signalling pathways in human myometrial cells in vitro by immunoblotting. Long-term treatment of myometrial cells with a series of concentrations of P4 and E2 in combination caused a change in the phosphorylation status of p42/44 mitogen activated protein kinase and of c-Jun N-terminal kinase (SAPK/JNK). P4 and E2 caused a decrease in protein expression of Gqalpha, Gzalpha, Gi1/2alpha and, to a lesser extent, G0alpha. The two steroids caused a decrease in the expression of the two small GSalpha isoforms. Cyclo-oxygenase-2 expression was increased by 2.5 fold after steroid treatment, while proliferating cell nuclear antigen expression levels remained unchanged. These observations show that the combination of P4 and E2 influences intracellular and membrane-bound components of signal transduction pathways in human myometrial cells. The implications of the two steroid hormones on intracellular signalling pathways in the human myometrium merits further investigation. PMID- 12087074 TI - Prolactin receptor expression in human testis and accessory tissues: localization and function. AB - Experimental studies in animals have established prolactin (PRL) as a progonadal hormone that promotes the function of the testis and reproductive accessory glands. The present study investigated the localization of PRL receptor (PRL-R) expression in the human testis and accessory tissues. Expression of PRL-R was identified in human testis and vas deferens by RT-PCR, and further localized by immunohistochemistry to the Leydig cells and differentiating germ cells of the testis (developmental stages extending from pachytene spermatocytes to elongating spermatids). Positive staining for PRL-R was also clearly evident in the epithelium of vas deferens, epididymis, prostate and seminal vesicles. Functional activation of PRL-R was demonstrated in fresh samples of vas deferens collected at vasectomy by examination of the JAK/STAT (Janus kinase/signal transducer and activator of transcription) and MAP (mitogen-activated protein) kinase ERK (extracellular signal-regulated kinase) signalling pathways. Within the vas deferens, PRL induced rapid tyrosine phosphorylation of JAK 2 and STAT 5 (after 10 and 20 min respectively), and tyrosine and threonine phosphorylation of ERK 1 and 2 (after 5 min). The demonstration of function and localization of PRL-R presented here suggests multiple roles for PRL in the human male reproductive tract. PMID- 12087075 TI - FSH and LH induce progesterone production and progesterone receptor synthesis in cumulus cells: a requirement for meiotic resumption in porcine oocytes. AB - The aim of this study was to investigate the role of progesterone in the meiotic resumption of porcine oocytes. Progesterone production and progesterone receptor (PR) immunoreactivity in cumulus cells were not detected in porcine cumulus oocyte complexes (COC) when observations were made either just after collection from the follicles or after 28 h cultivation without LH and FSH. However, the addition of LH and FSH induced PR expression in cumulus cells, concomitant with increased progesterone production. To assess the role of progesterone in the COC, an inhibitor of progesterone production, aminoglutethimide (AGT), was administered. The addition of AGT to the medium with LH and FSH significantly suppressed progesterone production in a dose-dependent fashion. When COC were cultured with LH, FSH and 0.5 x 10(-3) mol/l AGT, almost complete inhibition of progesterone production and of germinal vesicle breakdown (GVBD) was seen. However, this inhibitory effect on GVBD was overcome by additional progesterone. Moreover, 0.5 x 10(-3) mol/l AGT also suppressed the reduction in connexin43, a gap junctional protein, in cumulus cells after 28 h cultivation, and increased the level of cyclic AMP in oocytes. These results support the hypothesis that the binding of progesterone, which was secreted by LH- and FSH-stimulated cumulus cells, to its newly synthesized receptor induces GVBD in porcine oocytes, possibly through a reduction of connexin43 in cumulus cells. PMID- 12087076 TI - Modifications of the Ca2+ release mechanisms of mouse oocytes by fertilization and by sperm factor. AB - A cytosolic factor from sperm (SF) is thought to be responsible for the generation of intracellular calcium oscillations ([Ca2+]i) associated with fertilization in mammalian oocytes. Whether or not mouse oocytes injected with SF exhibit modifications of their Ca2+ release mechanisms similar to those observed in fertilized oocytes is not known and this was investigated here by injecting porcine SF (pSF). First, pSF-activated oocytes injected with CaCl2 showed persistent sensitization of the Ca2+-induced Ca2+ release mechanism, but this sensitization was absent in SrCl2-activated oocytes. Second, pSF-injected oocytes re-initiated oscillations when fused with untreated oocytes, although the Ca2+ responses were short-lived compared to those initiated by fertilization. Likewise, in the presence of colcemid, pSF-initiated oscillations were prolonged but ceased in advance of those in fertilized zygotes. Also, pronuclear envelope breakdown induced by okadaic acid was not associated with Ca2+ release in pSF generated zygotes, whereas it was observed in fertilized zygotes. Finally, roscovitine, an inhibitor of maturation promoting factor, blocked pSF-induced [Ca2+]i oscillations. Together, these results show that pSF-induced [Ca2+]i responses exhibit properties similar to those triggered by the sperm, although the SF's Ca2+ active component(s) may be less stable or more susceptible to degradation, resulting in shorter modification of the oocyte's Ca2+ release mechanisms. PMID- 12087077 TI - Epigenetic marks at BRCA1 and p53 coding sequences in early human embryogenesis. AB - In the vertebrate genome, methylation of deoxycytosine residues of CpGs dinucleotide has been associated with transcriptional silencing of genes, parental imprinting, X-inactivation and chromatin remodelling. In human somatic tissues, the 5' end of the BRCA1 CpG island is methylated, whereas this region is unmethylated in mature germ cells and early embryos. In gametes, as in somatic tissues, the CpG sites in the coding region are methylated. We took advantage of this bimodal distribution as a model to analyse the epigenetic reprogramming of coding regions during early human embryogenesis using the bisulphite-based genomic sequencing method. During preimplantation divisions, exon 11 of BRCA1 was slowly demethylated and retained approximately 30% of its methylated residues at the blastocyst stage. Moreover, the change in the distribution of methylated residues was not restricted to the BRCA1 gene, since for another gene, p53, a relatively high level of methylation (50%) of exon 4 was observed in blastocysts. Taken together, these data suggest that a significant part of the methylated residues of coding sequences might be conserved during preimplantation development. PMID- 12087078 TI - Interleukin 11 advances progesterone-induced decidualization of human endometrial stromal cells. AB - Differentiation of endometrial stromal cells into decidual cells is crucial for embryo implantation and placentation. Interleukin (IL)-11 signalling is essential for adequate decidualization in the mouse uterus. We examined the role of IL-11 during progesterone-induced decidualization of human endometrial stromal cells over a 10-12 day period, using prolactin (PRL) production as a decidual marker. These cells produced biologically active IL-11 and expressed IL-11, IL-11Ralpha and PRL mRNA during decidualization. Neutralization of endogenous IL-11 with an anti-human (hu)IL-11 antibody (AB) reduced production of PRL from day 8 and insulin-like growth factor binding protein (IGFBP)-1, another marker of decidualization, from day 10 of culture. Following AB washout, PRL and IGFBP-1 secretion increased. Addition of recombinant (r)huIL-11 (10 or 100 ng/ml) to endometrial stromal cells increased secretion of PRL from day 4 and IGFBP-1 from day 6 compared with progesterone alone. Morphological signs of differentiation accompanied biochemical differentiation in the progesterone-treated cells and were further induced by exogenous rhuIL-11. Our observations demonstrate that human endometrial stromal cells produce biologically active IL-11, which promotes progesterone-induced decidualization. These results suggest that IL-11 has both paracrine and autocrine actions on human endometrial stromal cells and plays an important role in preparing the human endometrium for implantation. PMID- 12087079 TI - Endometrial nuclear receptor co-factors SRC-1 and N-CoR are increased in human endometrium during menstruation. AB - Steroid hormone receptor co-factors are abundantly expressed in the uterus in order to modify steroid hormone receptor action, either leading to activation or repression of transcription in the endometrium. However, the role of co-factors in remodelling of the human endometrium has not been established. We therefore endeavoured to evaluate the presence of the co-activator SRC (steroid receptor co activator)-1 and the co-repressors N-CoR (nuclear receptor co-repressor) and steroid co-repressor SMRT (silencing mediator of retinod and thyroid) receptors in the human endometrium during the different phases of the menstrual cycle. By using a real-time RT-PCR assay, we showed that SRC-1, N-CoR and SMRT mRNA are expressed in human endometrium during all phases of the menstrual cycle, as well as in inactive endometrium. Moreover, endometrial expression of SRC-1 and N-CoR mRNA increased during menstruation when compared with the other phases of the menstrual cycle (P < 0.001). Immunohistochemistry demonstrated that SRC-1 and N CoR stain positive in the glandular epithelium and stroma in menstrual phase endometrium. The staining was weak in proliferative and secretory endometrium and absent in inactive endometrium. Our results suggest that differential expression of endometrial steroid receptor co-factors probably play a role in the regulation of human endometrium remodelling. PMID- 12087080 TI - Expression and regulation of the fatty acid amide hydrolase gene in the rat uterus during the estrous cycle and peri-implantation period. AB - Fatty acid amide hydrolase (FAAH) is involved in embryo development and implantation. Sex hormones down-modulate FAAH activity in the mouse uterus. However, the regulation of the FAAH gene in the uterus is unknown. Our results showed that FAAH mRNA is localized to uterine epithelial cells and circular myometrium during the estrous cycle. In ovariectomized rats, estradiol (E2) plus progesterone (P4) increased FAAH levels in both epithelial cells and circular myometrium. Interestingly, during the implantation period, FAAH mRNA was detected not only in epithelial cells and circular myometrium, but also in the primary decidual zone surrounding the implanting embryo on day 6 and in whole decidualized stromal cells on day 7. Its levels in the stromal cells were markedly higher at the implantation sites than at the inter-implantation sites on days 6 and 7. When implantation was delayed and then induced by E2 or E2 plus P4, FAAH mRNA levels were significantly increased in subepithelial stromal cells and circular myometrium, indicating that blastocyst activation and initiation of implantation in rats requires higher expression of the FAAH gene in subepithelial stromal cells and circular myometrium. In conclusion, the expression of FAAH mRNA is different in the non-pregnant and pregnant rat uterus and sex hormones up regulate FAAH gene expression. PMID- 12087081 TI - Expression of matrix metalloproteinase-26 and tissue inhibitor of metalloproteinase-4 in human normal cytotrophoblast cells and a choriocarcinoma cell line, JEG-3. AB - The balance between matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) is critical for embryo implantation. Disturbance of this balance may lead to tumour metastasis. To understand the roles of MMP-26 and TIMP-4 in physiological and pathological invasion, the expression of these proteins in normal human cytotrophoblast cells and in a malignant choriocarcinoma cell line, JEG-3, was investigated. MMP-26 and TIMP-4 proteins were detected in the cytoplasm of these cells. The expression levels of MMP-26 mRNA and protein in JEG-3 cells were significantly higher than those in the cytotrophoblasts; conversely, the expression levels of TIMP-4 mRNA and protein were much lower in JEG-3 cells than those in cytotrophoblasts (P < 0.01). Enzyme inhibition studies demonstrated that TIMP-4 was a potent inhibitor of MMP-26 with an IC50 value of 0.4 nmol/l. This study confirms that MMP-26 is an epithelial enzyme and suggests that MMP-26 and TIMP-4 may play a role in tissue-remodelling processes associated with placentation and tumour progression, and that a higher MMP-26 to TIMP-4 ratio may promote cancer invasion. PMID- 12087082 TI - Expression of the interleukin-2 receptor alpha (CD25) is selectively decreased on decidual CD4+ and CD8+ T lymphocytes in normal pregnancies. AB - In a previous study, we demonstrated that the proportion of activated T cells (CD69+CD3+ and HLA-DR+CD3+) is higher in the endometrium and decidua after the luteal phase and throughout early pregnancy compared with in the peripheral blood. However, there was no difference in the proportion of CD25+CD3+ lymphocytes between the endometrium and peripheral blood. In this study, we further verify that the levels of CD25 on CD4+ and CD8+ T lymphocytes are not increased in normal pregnancy, although the levels of CD69 and HLA-DR are markedly increased. We also elucidate that the amounts of all three activation molecules on local T lymphocytes are down-regulated in pregnancy compared with that during the luteal phase. Nevertheless, these decreases are significantly lessened in anembryonic pregnancies with both normal and abnormal karyotyping. However, in peripheral blood, the down-regulation of activation molecules levels in pregnancy is only demonstrated on CD4+ cells and for HLA-DR on CD8+ cells. Furthermore, dual activation marker analysis demonstrated that the expression of CD25 appears to be dissociated from CD69 and HLA-DR on the same decidual lymphocytes. Because IL-2Ralpha plays a pivotal role in the development and propagation of functional T cells, its depressed expression may result in maternal tolerance of the fetal allograft. PMID- 12087083 TI - Microarray analysis of differentially expressed genes in placental tissue of pre eclampsia: up-regulation of obesity-related genes. AB - Susceptibility genes present in both mother and fetus most likely contribute to the risk of pre-eclampsia. Placental biopsies were therefore investigated by high density DNA microarray analysis to determine genes differentially regulated within chorionic villous tissue in pre-eclampsia and normal pregnancy. The pooled RNAs of pre-eclamptic and normotensive subjects were hybridized to the HuGeneFL array representing sequences from approximately 5600 full-length human cDNAs. The differentially expressed genes that were detected could be categorized into nine groups: adhesion molecules, obesity-related genes, transcription factors/signalling molecules, immunological factors, neuromediators, oncogenic factors, protease inhibitors, hormones and growth factor-binding proteins. Among those, the obesity-related genes included putative candidate genes associated with the pathogenesis of pre-eclampsia. One of the most up-regulated transcripts was the obese gene (43.6-fold change), and this was reflected by elevated leptin protein levels. In the case of feto-maternal contribution of polymorphic genes to pre-eclampsia, expression analysis of placental tissue has lead to numerous target genes waiting for large scale genetic linkage analyses. PMID- 12087084 TI - Prostaglandin F(2alpha), cytokines and cyclic mechanical stretch augment matrix metalloproteinase-1 secretion from cultured human uterine cervical fibroblast cells. AB - Human uterine cervical tissue is composed mainly of fibroblast cells and the extracellular matrix in which collagen types I and III predominate. It is hypothesized that these collagens are degraded by matrix metalloproteinases (MMPs) in the initial step of uterine cervical ripening during parturition. Among the MMPs, MMP-1, -8 and -13 have substrate selectivity for collagen types I and III. In the present study, we examined the regulation of MMP-1 secretion from the human uterine cervix. Immunohistochemistry detected strong staining of MMP-1, but not of MMP-8 or -13, in stromal cells of the pregnant uterine cervix. The MMP-1 expression in the pregnant uterine cervix was further confirmed by Western blot analysis and RT-PCR. To clarify the regulation of MMP-1 production, we subsequently investigated the effects of prostaglandins, inflammatory cytokines and cyclic mechanical stretch on the secretion of MMP-1 from cultured human uterine cervical fibroblast cells. Treatment with prostaglandin (PG)F(2alpha) (10(-7) to 10(-5) mol/l) or interleukin (IL)-1alpha (0.01-1.0 ng/ml) or stimulation with cyclic mechanical stretch increased MMP-1 secretion from cultured human uterine cervical fibroblast cells, with maximal increases of 3.4-, 4.5- and 1.9-fold respectively (24 h of treatment, P < 0.05 for all comparisons). These data suggest that MMP-1 may play a significant role in the degradation of extracellular collagen types I and III in the pregnant uterine cervix during the process of cervical ripening, in response to various stimulations such as PGF(2alpha), IL-1alpha and mechanical stretch. PMID- 12087085 TI - Birth of healthy female twins after preimplantation genetic diagnosis of cystic fibrosis combined with gender determination. AB - Two healthy sisters with a familial history of mental retardation were referred to our centre for preimplantation genetic diagnosis (PGD). Their two brothers showed severe mental retardation. The molecular basis for their disorder could not be identified, but one of the sisters and the mother presented a highly skewed pattern of X-inactivation reinforcing the likelihood of an X-linked mode of inheritance. Both sisters requested PGD to avoid the abortion of potentially affected male fetuses. PGD for sex by fluorescent in-situ hybridization was carried out for the first sister and resulted in the birth of a female child. The second sister and her partner, whose niece had cystic fibrosis (CF), were tested for CF mutations, and were both found to be deltaF508 heterozygous. We developed an efficient single cell PCR protocol for the simultaneous amplification of the CF (deltadeltaF508) locus as well as the X-linked amelogenin gene and its highly homologous pseudogene on the Y chromosome. Two PGD cycles were carried out to screen against male and deltaF508 homozygous deleted embryos. In each case several embryos could be selected for transfer and the second cycle resulted in a twin pregnancy followed by the birth of two healthy female infants. PMID- 12087086 TI - Oligomerization state-dependent activation of NF-kappa B signaling pathway by adipocyte complement-related protein of 30 kDa (Acrp30). AB - Adipocyte complement-related protein of 30 kDa (Acrp30)/adiponectin is an adipocyte-derived hormone that affects lipid and glucose metabolism in muscle and liver, but its physical and biochemical properties are poorly characterized. Here we have used several approaches to show that Acrp30 expressed in and purified from Escherichia coli and human embryonic kidney 293T cells forms trimers and hexamers; 293T cells also produce a higher molecular weight species. Similar Acrp30 oligomers were found in mouse serum as well as in 3T3-L1 adipocyte conditioned medium, although in different proportions. In parallel, we assessed whether Acrp30 is a signaling molecule by searching for promoter or enhancer elements that respond to Acrp30 or its isolated trimeric globular C-terminal domain, gAcrp30. Acrp30 addition to C2C12 myocytes or myotubes led to activation of NF-kappa B transcription factor in a manner dependent upon phosphorylation and degradation of I kappa B-alpha. Importantly, only hexameric and larger isoforms of Acrp30 activated NF-kappa B; trimeric Acrp30 or gAcrp30 could not activate NF kappa B. Our data indicate that oligomerization of Acrp30 is important for at least some of its biological activities, and changes in the relative abundance of each oligomeric isoform in plasma may regulate Acrp30 activity. PMID- 12087087 TI - RNA polymerase II transcription complexes may become arrested if the nascent RNA is shortened to less than 50 nucleotides. AB - A significant fraction of RNA polymerase II transcription complexes become arrested when halted within a particular initially transcribed region after the synthesis of 23-32-nucleotide RNAs. If polymerases are halted within the same sequence at a promoter-distal location, they remain elongation-competent. However, when the RNAs within these promoter-distal complexes are truncated to between 21 and 48 nucleotides, many of the polymerases become arrested. The degree of the arrest correlates very well with the length of the RNA in both the promoter-proximal and -distal complexes. This effect is also observed when comparing promoter-proximal and promoter-distal complexes halted over a completely different sequence. The unusual propensity of many promoter-proximal RNA polymerase II complexes to arrest may therefore be recreated in promoter distal complexes simply by shortening the nascent RNA. Thus, the transition to full elongation competence by RNA polymerase II is dependent on the synthesis of about 50 nt of RNA, and this transition is reversible. We also found that arrest is facilitated in promoter-distal complexes by the hybridization of oligonucleotides to the transcript between 30 and 45 bases upstream of the 3' end. PMID- 12087088 TI - Syndecan-4 modulates focal adhesion kinase phosphorylation. AB - The cell-surface heparan sulfate proteoglycan syndecan-4 acts in conjunction with the alpha(5)beta(1) integrin to promote the formation of actin stress fibers and focal adhesions in fibronectin (FN)-adherent cells. Fibroblasts seeded onto the cell-binding domain (CBD) fragment of FN attach but do not fully spread or form focal adhesions. Activation of Rho, with lysophosphatidic acid (LPA), or protein kinase C, using the phorbol ester phorbol 12-myristate 13-acetate, or clustering of syndecan-4 with antibodies directed against its extracellular domain will stimulate formation of focal adhesions and stress fibers in CBD-adherent fibroblasts. The distinct morphological differences between the cells adherent to the CBD and to full-length FN suggest that syndecan-4 may influence the organization of the focal adhesion or the activation state of the proteins that comprise it. FN-null fibroblasts (which express syndecan-4) exhibit reduced phosphorylation of focal adhesion kinase (FAK) tyrosine 397 (Tyr(397)) when adherent to CBD compared with FN-adherent cells. Treating the CBD-adherent fibroblasts with LPA, to activate Rho, or the tyrosine phosphatase inhibitor sodium vanadate increased the level of phosphorylation of Tyr(397) to match that of cells plated on FN. Treatment of the fibroblasts with PMA did not elicit such an effect. To confirm that this regulatory pathway includes syndecan-4 specifically, we examined fibroblasts derived from syndecan-4-null mice. The phosphorylation levels of FAK Tyr(397) were lower in FN-adherent syndecan-4-null fibroblasts compared with syndecan-4-wild type and these levels were rescued by the addition of LPA or re-expression of syndecan-4. These data indicate that syndecan-4 ligation regulates the phosphorylation of FAK Tyr(397) and that this mechanism is dependent on Rho but not protein kinase C activation. In addition, the data suggest that this pathway includes the negative regulation of a protein tyrosine phosphatase. Our results implicate syndecan-4 activation in a direct role in focal adhesion regulation. PMID- 12087089 TI - ChREBP rather than USF2 regulates glucose stimulation of endogenous L-pyruvate kinase expression in insulin-secreting cells. AB - There is controversy whether or not upstream stimulatory factors (USF) regulate the glucose responsiveness of L-pyruvate kinase (L-PK) promoter activity in hepatocytes. It has been suggested that USF-2 is required for glucose stimulation of L-PK promoter activity in single islet beta-cells and INS-1 cells (Kennedy, H. J., Viollet, B., Rafiq, I., Kahn, A., and Rutter, G. A. (1997) J. Biol. Chem. 272, 20636-20640). In the present study, the tet-on system has been employed to achieve tightly controlled and inducible expression of USF-1 and -2 and their dominant-negative mutants DN-USF-1 (DeltabTDU1) and -2 (TDU2) in INS-1 cells. Quantitative Northern blot analysis shows that neither basal level nor glucose responsiveness of endogenous L-PK mRNA is affected by overexpression of USF-1 and -2. Likewise, the L-PK expression is unaltered by dominant-negative suppression of USF function. Western blotting demonstrates that USF-1 and -2 and DN-USF-1 and -2 proteins are stably expressed in nuclear fractions of INS-1 cells. Immunofluorescence staining indicates the uniform induction of these transgene encoded proteins in the cell nuclei. Electrophoretic mobility shift assays using the L-PK promoter segment reveal that induction of USF-1 and -2 dramatically enhances the USF binding activity, whereas DN-USF-1 and -2 abolish binding. DN USF-1 and -2 exert their dominant-negative effect by forming non-functional heterodimers with endogenous USF proteins. Carbohydrate response element-binding protein (ChREBP) was recently shown to regulate the glucose responsiveness of the L-PK promoter activity in hepatocytes. We now report the presence of this transcription factor in rat islets and INS-1 cells. Glucose stimulates ChREBP transcription in INS-1 cells, as shown by nuclear run-on experiments. Overexpression of ChREBP in INS-1 cells using the tet-on system results in a left shift of glucose responsiveness of L-PK expression and an enhanced L-PK promoter activity. Both endogenous and doxycycline-induced ChREBP proteins bind to the L PK promoter in a glucose-dependent manner. These unprecedented results suggest that ChREBP rather than USF mediates glucose-promoted L-PK expression in insulin secreting cells. PMID- 12087090 TI - Long CTG.CAG repeats from myotonic dystrophy are preferred sites for intermolecular recombination. AB - Homologous recombination was shown to enable the expansion of CTG.CAG repeat sequences. Other prior investigations revealed the involvement of replication and DNA repair in these genetic instabilities. Here we used a genetic assay to measure the frequency of homologous intermolecular recombination between two CTG.CAG tracts. When compared with non-repeating sequences of similar lengths, long (CTG.CAG)(n) repeats apparently recombine with an approximately 60-fold higher frequency. Sequence polymorphisms that interrupt the homogeneity of the CTG.CAG repeat tracts reduce the apparent recombination frequency as compared with the pure uninterrupted repeats. The orientation of the repeats relative to the origin of replication strongly influenced the apparent frequency of recombination. This suggests the involvement of DNA replication in the recombination process of triplet repeats. We propose that DNA polymerases stall within the CTG.CAG repeat tracts causing nicks or double-strand breaks that stimulate homologous recombination. The recombination process is RecA-dependent. PMID- 12087091 TI - Nonsteroidal anti-inflammatory drugs inhibit matrix metalloproteinase-2 via suppression of the ERK/Sp1-mediated transcription. AB - Our previous data showed that nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit matrix metalloproteinase-2 (MMP-2) expression via repression of gene transcription in lung cancer cells. In this study, we investigate the molecular mechanism by which NSAIDs inhibit MMP-2. Promoter deletion and mutation analysis indicate that NSAIDs act via the Sp1 transcription factor binding site located between -91 and -84 in the MMP-2 promoter to suppress gene expression. Electrophoretic mobility shift assays show that Sp1 and Sp3 proteins constitutively bind to this consensus sequence and overexpression of Sp1 may enhance MMP-2 expression. NSAID treatment reduces Sp1 DNA binding activity and phosphorylation and attenuates MMP-2 expression. We also investigate the signaling pathway that mediates the effect of NSAIDs. Our results suggest that ERKs are involved in this process. First, NSAIDs suppress basal and serum stimulated ERK activity. Second, a MEK inhibitor PD98059 inhibits MMP-2 promoter activity and Sp1 phosphorylation. Third, overexpression of constitutively active MEK1 stimulates Sp1 phosphorylation and MMP-2 promoter activity and antagonizes the inhibition of NSAIDs. Collectively, our data suggest that NSAIDs inhibit MMP 2 by blocking ERK/Sp1-mediated transcription. PMID- 12087093 TI - Cobalt substitution of mouse R2 ribonucleotide reductase as a model for the reactive diferrous state: spectroscopic and structural evidence for a ferromagnetically coupled dinuclear cobalt cluster. AB - The R2 dimer of mouse ribonucleotide reductase contains a dinuclear iron-oxygen cluster and tyrosyl radical/subunit. The dinuclear diferrous form reacts with dioxygen to generate the tyrosyl radical essential for the catalytic reaction that occurs at the R1 dimer. It is important to understand how the reactivity toward oxygen is related to the crystal structure of the dinuclear cluster. For the mouse R2 protein, no structure has been available with a fully occupied dinuclear metal ion site. A cobalt substitution of mouse R2 was performed to produce a good model for the very air-sensitive diferrous form of the enzyme. X band EPR and light absorption studies (epsilon(550 nm) = 100 mm(-1) cm( 1)/Co(II)) revealed a strong cooperative binding of cobalt to the dinuclear site. In perpendicular mode EPR, the axial signal from mouse R2 incubated with Co(II) showed a typical S = 3/2 Co(II) signal, and its low intensity indicated that the majority of the Co(II) bound to R2 is magnetically coupled. In parallel mode EPR, a typical integer spin signal (M(s) = +/-3) with g approximately 12 is observed at 3.6 K and 10 K, showing that the two Co(II) ions (S = 3/2) in the dinuclear site are ferromagnetically coupled. We have solved the 2.4 A crystal structure of the Co(II)-substituted R2 with a fully occupied dinuclear cluster. The bridging Co(II) carboxylate ligand Glu-267 adopts an altered orientation compared with its counterpart Glu-238 in Escherichia coli R2. This might be important for proper O(2) activation of the more exposed native diferrous site in mouse R2 compared with E. coli R2. PMID- 12087092 TI - Role of Dok1 in cell signaling mediated by RET tyrosine kinase. AB - Using a yeast two-hybrid screen, we identified Dok1 as a docking protein for RET tyrosine kinase. Dok1 bound more strongly to RET with a multiple endocrine neoplasia (MEN) 2B mutation than RET with a MEN2A mutation and was highly phosphorylated in the cells expressing the former mutant protein. Analysis by site-directed mutagenesis revealed that tyrosine 361 in mouse Dok1 represents a binding site for the Nck adaptor protein and tyrosines 295, 314, 361, 376, 397, and 408 for the Ras-GTPase-activating protein. We replaced tyrosine 361 or these six tyrosines with phenylalanine (designated Y361F or 6F) in Dok1 and introduced the mutant Dok1 genes into the cells expressing the wild-type RET or RET-MEN2B protein. Overexpression of Dok1 or Dok1-Y361F, but not Dok1-6F, suppressed the Ras/Erk activation induced by glial cell line-derived neurotrophic factor or RET MEN2B, implying that this inhibitory effect requires the Ras-GTPase-activating protein binding to Dok1. In contrast, overexpression of Dok1, but not Dok1-Y361F or Dok1-6F, enhanced the c-Jun amino-terminal kinase (JNK) and c-Jun activation. This suggested that the association of Nck to tyrosine 361 in Dok1 is necessary for the JNK and c-Jun activation by glial cell line-derived neurotrophic factor or RET-MEN2B. Because a high level of the JNK phosphorylation was observed in the cells expressing RET-MEN2B, its strong activation via Nck binding to Dok1 may be responsible for aggressive properties of medullary thyroid carcinoma developed in MEN 2B. PMID- 12087094 TI - Cloning and characterization of Ehox, a novel homeobox gene essential for embryonic stem cell differentiation. AB - We report here the identification and characterization of a novel paired-like homeobox-containing gene (Ehox). This gene, identified in embryonic stem (ES) cells, is differentially expressed during in vitro ES cell differentiation. We have assessed Ehox function using the ES cell in vitro differentiation system. This has involved molecular and biological analyses of the effects of sense or antisense Ehox expression (using episomal vectors) on ES cell differentiation. Analysis of antisense Ehox-expressing ES cells indicates that they are unable to express marker genes associated with hematopoietic, endothelial, or cardiac differentiation following removal of leukemia inhibitory factor. In contrast, overexpression of Ehox using the sense construct accelerated the appearance of these differentiation markers. ES cell self-renewal and differentiation assays reveal that inhibition of Ehox activity results in the maintenance of a stem cell phenotype in limiting concentrations of leukemia inhibitory factor and the almost complete impairment of the cardiomyocyte differentiation capacity of these cells. We therefore conclude that Ehox is a novel homeobox-containing gene that is essential for the earliest stages of murine ES cell differentiation. PMID- 12087096 TI - Tubulin is an inherent component of mitochondrial membranes that interacts with the voltage-dependent anion channel. AB - We have previously reported that anti-tubulin agents induce the release of cytochrome c from isolated mitochondria. In this study, we show that tubulin is present in mitochondria isolated from different human cancerous and non-cancerous cell lines. The absence of polymerized microtubules and cytosolic proteins was checked to ensure that this tubulin is an inherent component of the mitochondria. In addition, a salt wash did not release the tubulin from the mitochondria. By using electron microscopy, we then showed that tubulin is localized in the mitochondrial membranes. As compared with cellular tubulin, mitochondrial tubulin is enriched in acetylated and tyrosinated alpha-tubulin and is also enriched in the class III beta-tubulin isotype but contains very little of the class IV beta tubulin isotype. The mitochondrial tubulin is likely to be organized in alpha/beta dimers and represents 2.2 +/- 0.5% of total cellular tubulin. Lastly, we showed by immunoprecipitation experiments that the mitochondrial tubulin is specifically associated with the voltage-dependent anion channel, the main component of the permeability transition pore. Thus, tubulin is an inherent component of mitochondrial membranes, and it could play a role in apoptosis via interaction with the permeability transition pore. PMID- 12087097 TI - Inhibition of phosphorylation of BAD and Raf-1 by Akt sensitizes human ovarian cancer cells to paclitaxel. AB - We studied the roles of the phosphatidylinositol 3-kinase (PI-3K)-Akt-BAD cascade, ERK-BAD cascade, and Akt-Raf-1 cascade in the paclitaxel-resistant SW626 human ovarian cancer cell line, which lacks functional p53. Treatment of SW626 cells with paclitaxel activates Akt and ERK with different time frames. Interference with the Akt cascade either by treatment with PI-3K inhibitor (wortmannin or LY294002) or by exogenous expression of a dominant negative Akt in SW626 cells caused decreased cell viability following treatment with paclitaxel. Interference with the ERK cascade by treatment with an MEK inhibitor, PD98059, in SW626 cells also caused decreased cell viability following treatment with paclitaxel. Treatment of cells with paclitaxel also stimulated the phosphorylation of BAD at both the Ser-112 and Ser-136 sites. The phosphorylation of BAD at Ser-136 was blocked by treatment with wortmannin or cotransfection with the dominant negative Akt. On the other hand, the phosphorylation of BAD at Ser 112 was blocked by PD98059. We further examined the role of BAD in the viability following paclitaxel treatment using BAD mutants. Exogenous expression of doubly substituted BAD2SA in SW626 cells caused decreased viability following treatment with paclitaxel. Moreover, because paclitaxel-induced apoptosis is mediated by activated Raf-1 and the region surrounding Ser-259 in Raf-1 conforms to a consensus sequence for phosphorylation by Akt, the regulation of Raf-1 by Akt was examined. We demonstrated an association between Akt and Raf-1 and showed that the phosphorylation of Raf-1 on Ser-259 induced by paclitaxel was blocked by treatment with wortmannin or LY294002. Furthermore, interference with the Akt cascade induced by paclitaxel up-regulated Raf-1 activity, and expression of constitutively active Akt inhibited Raf-1 activity, suggesting that Akt negatively regulates Raf-1. Our findings suggest that paclitaxel induces the phosphorylation of BAD Ser-112 via the ERK cascade, and the phosphorylation of both BAD Ser-136 and Raf-1 Ser-259 via the PI-3K-Akt cascade, and that inhibition of either of these cascades sensitizes ovarian cancer cells to paclitaxel. PMID- 12087095 TI - Bacterial surface association of heat-labile enterotoxin through lipopolysaccharide after secretion via the general secretory pathway. AB - Heat-labile enterotoxin (LT) is an important virulence factor expressed by enterotoxigenic Escherichia coli. The route of LT secretion through the outer membrane and the cellular and extracellular localization of secreted LT were examined. Using a fluorescently labeled receptor, LT was found to be specifically secreted onto the surface of wild type enterotoxigenic Escherichia coli. The main terminal branch of the general secretory pathway (GSP) was necessary and sufficient to localize LT to the bacterial surface in a K-12 strain. LT is a heteromeric toxin, and we determined that its cell surface localization was mediated by the its B subunit independent of an intact G(M1) ganglioside binding site and that LT binds lipopolysaccharide and G(M1) concurrently. The majority of LT secreted into the culture supernatant by the GSP in E. coli associated with vesicles. Only a mutation in hns, not overexpression of the GSP or LT, caused an increase in vesicle yield, supporting a specific vesicle formation machinery regulated by the nucleoid-associated protein HNS. We propose a model in which LT is secreted by the GSP across the outer membrane, secreted LT binds lipopolysaccharide via a G(M1)-independent binding region on its B subunit, and LT on the surface of released outer membrane vesicles interacts with host cell receptors, leading to intoxication. These data explain a novel mechanism of vesicle-mediated receptor-dependent delivery of a bacterial toxin into a host cell. PMID- 12087098 TI - Regulation of ribosomal S6 kinase 2 by mammalian target of rapamycin. AB - Phosphorylation of the ribosomal S6 subunit is tightly correlated with enhanced translation initiation of a subset of mRNAs that encodes components of the protein synthesis machinery, which is an important early event that controls mammalian cell growth and proliferation. The recently identified S6 kinase 2 (S6K2), together with its homologue S6K1, is likely responsible for the mitogen stimulated phosphorylation of S6. Like S6K1, the activation of S6K2 requires signaling from both the phosphatidylinositol 3-kinase and the mammalian target of rapamycin (mTOR). Here we report the investigation of the mechanisms of S6K2 regulation by mTOR. We demonstrate that similar to S6K1 the serum activation of S6K2 in cells is dependent on mTOR kinase activity, amino acid sufficiency, and phosphatidic acid. Previously we have shown that mTOR is a cytoplasmic-nuclear shuttling protein. As a predominantly nuclear protein, S6K2 activation was facilitated by enhanced mTOR nuclear import with the tagging of an exogenous nuclear localization signal and diminished by enhanced mTOR nuclear export with the tagging of a nuclear export sequence. However, further increase of mTOR nuclear import by the tagging of four copies of nuclear localization signal resulted in its decreased ability to activate S6K2, suggesting that mTOR nuclear export may also be an integral part of the activation process. Consistently, the nuclear export inhibitor leptomycin B inhibited S6K2 activation. Taken together, our observations suggest a novel regulatory mechanism in which an optimal cytoplasmic-nuclear distribution or shuttling rate for mTOR is required for maximal activation of the nuclear S6K2. PMID- 12087099 TI - The proton channel of the energy-transducing nicotinamide nucleotide transhydrogenase of Escherichia coli. AB - The nicotinamide nucleotide transhydrogenases of mitochondria and bacteria are proton pumps that couple direct hydride ion transfer between NAD(H) and NADP(H) bound, respectively, to extramembranous domains I and III to proton translocation by the membrane-intercalated domain II. To delineate the proton channel of the enzyme, 25 conserved and semiconserved prototropic amino acid residues of domain II of the Escherichia coli transhydrogenase were mutated, and the mutant enzymes were assayed for transhydrogenation from NADPH to an NAD analogue and for the coupled outward proton translocation. The results confirmed the previous findings of others and ourselves on the essential roles of three amino acid residues and identified another essential residue. Three of these amino acids, His-91, Ser 139, and Asn-222, occur in three separate membrane-spanning alpha helices of domain II of the beta subunit of the enzyme. Another residue, Asp-213, is probably located in a cytosolic-side loop that connects to the alpha helix bearing Asn-222. It is proposed that the three helices bearing His-91, Ser-139, and Asn-222 come together, possibly with another highly conserved alpha helix to form a four-helix bundle proton channel and that Asp-213 serves to conduct protons between the channel and domain III where NADPH binding energy is used via protein conformation change to initiate outward proton translocation. PMID- 12087100 TI - Interleukin-22 (IL-22) activates the JAK/STAT, ERK, JNK, and p38 MAP kinase pathways in a rat hepatoma cell line. Pathways that are shared with and distinct from IL-10. AB - IL (interleukin)-22 is an IL-10-related cytokine; its main biological activity known thus far is the induction of acute phase reactants in liver and pancreas. IL-22 signals through a receptor that is composed of two chains from the class II cytokine receptor family: IL-22R (also called ZcytoR11/CRF2-9) and IL-10Rbeta (CRF2-4), which is also involved in IL-10 signaling. In this report, we analyzed the signal transduction pathways activated in response to IL-22 in a rat hepatoma cell line, H4IIE. We found that IL-22 induces activation of JAK1 and Tyk2 but not JAK2, as well as phosphorylation of STAT1, STAT3, and STAT5 on tyrosine residues, extending the similarities between IL-22 and IL-10. However our results unraveled some differences between IL-22 and IL-10 signaling. Using antibodies specific for the phosphorylated form of MEK1/2, ERK1/2, p90RSK, JNK, and p38 kinase, we showed that IL-22 activates the three major MAPK pathways. IL-22 also induced serine phosphorylation of STAT3 on Ser(727). This effect, which is not shared with IL 10, was only marginally affected by MEK1/2 inhibitors, indicating that other pathways might be involved. Finally, by overexpressing a STAT3 S727A mutant, we showed that serine phosphorylation is required to achieve maximum transactivation of a STAT responsive promoter upon IL-22 stimulation. PMID- 12087102 TI - Protein determinants of RNA binding by DNA polymerase of the T4-related bacteriophage RB69. AB - DNA polymerase (gp43) of phage T4 plays two biological roles, one as an essential DNA binding replication enzyme and the other as an mRNA-specific autogenous translational repressor. Binding of T4 gp43 to its mRNA target (translational operator RNA) interferes with gp43-DNA interactions, but it is unclear how the protein determinants for binding DNA are affected by the dynamics of gp43-mRNA interactions. We have used RB69 gp43, a natural variant of the T4 enzyme whose crystal structure has been determined to identify protein sites that respond to the interaction with specific RNA. We used protein phosphorylation markers, photocross-linking studies, protease sensitivity assays, and mutational analyses to examine the effects of operator RNA on the enzyme's five structural domains (N, exo, palm, fingers, and thumb). Our studies suggest that this RNA affects gp43-DNA interactions through global effects on protein structure that occlude DNA-binding sites but leave the enzyme accessible to interactions with the sliding clamp (RB69 gp45) and possibly other polymerase accessory proteins. We discuss the possible biological significance of putative RNA-binding motifs in the N and palm domains of RB69 gp43. PMID- 12087101 TI - MCM2-7 complexes bind chromatin in a distributed pattern surrounding the origin recognition complex in Xenopus egg extracts. AB - The MCM2-7 complex is believed to function as the eukaryotic replicative DNA helicase. It is recruited to chromatin by the origin recognition complex (ORC), Cdc6, and Cdt1, and it is activated at the G(1)/S transition by Cdc45 and the protein kinases Cdc7 and Cdk2. Paradoxically, the number of chromatin-bound MCM complexes greatly exceeds the number of bound ORC complexes. To understand how the high MCM2-7:ORC ratio comes about, we examined the binding of these proteins to immobilized linear DNA fragments in Xenopus egg extracts. The minimum length of DNA required to recruit ORC and MCM2-7 was approximately 80 bp, and the MCM2 7:ORC ratio on this fragment was approximately 1:1. With longer DNA fragments, the MCM2-7:ORC ratio increased dramatically, indicating that MCM complexes normally become distributed over a large region of DNA surrounding ORC. Only a small subset of the chromatin-bound MCM2-7 complexes recruited Cdc45 at the onset of DNA replication, and unlike Cdc45, MCM2-7 was not limiting for DNA replication. However, all the chromatin-bound MCM complexes may be functional, because they were phosphorylated in a Cdc7-dependent fashion, and because they could be induced to support Cdk2-dependent Cdc45 loading. The data suggest that in Xenopus egg extracts, origins of replication contain multiple, distributed, initiation-competent MCM2-7 complexes. PMID- 12087103 TI - Correlation of transcriptional repression by p21(SNFT) with changes in DNA.NF-AT complex interactions. AB - p21(SNFT) (21-kDa small nuclear factor isolated from T cells) is a novel human protein of the basic leucine zipper family. The overexpression of p21(SNFT) leads to the significant and specific repression of transcription from the interleukin 2 promoter as well as from several essential activator protein 1 (AP-1)-driven composite promoter elements. One example is the distal nuclear factor of activated T cells (NF-AT)/AP-1 element where the AP-1 (Fos/Jun) basic leucine zipper heterodimer interacts with members of the NF-AT family. p21(SNFT) has been shown to replace Fos in dimerization with Jun on a consensus AP-1 binding site (12-O-tetradecanolyphorbol-13-acetate response element (TRE)) and to interact with Jun and NF-AT at the distal NF-AT/AP-1 enhancer element. A detailed biochemical analysis presented here compares interactions involving p21(SNFT) with those involving Fos. The results demonstrate that a p21(SNFT)/Jun dimer binds a TRE similarly to AP-1 and like AP-1 binds cooperatively with NF-AT at the NF-AT/AP-1 composite element. However, Fos interacts significantly more efficiently than p21(SNFT) with Jun and NF-AT, and the replacement of Fos by p21(SNFT) in the trimolecular complex drastically alters protein-DNA contacts. The data suggest that p21(SNFT) may repress transcriptional activity by inducing a unique conformation in the transcription factor complex. PMID- 12087104 TI - Calpain activity regulates the cell surface distribution of amyloid precursor protein. Inhibition of calpains enhances endosomal generation of beta-cleaved C terminal APP fragments. AB - In murine L cells, treatment with calpeptin or calpain inhibitor III increased Abeta42, but not Abeta40, secretion in a dose-dependent fashion. This correlated with an increase in the levels of amyloid precursor protein (APP) carboxyl terminal fragments (CTFs). Immunoprecipitation with novel mAbs directed against the carboxyl-terminus of APP or specific for the beta-cleaved CTF showed that generation of both alpha- and beta-cleaved CTFs increase proportionately following inhibition of calpains. Pulse-chase metabolic labeling confirmed that inhibiting calpains increases the production of alpha- and beta-cleaved APP metabolites. Immunolabeling showed greater betaCTF signal in calpeptin-treated cells, primarily in small vesicular compartments that were shown to be predominantly endosomal by colocalization with early endosomal antigen 1. A second mAb, which recognizes an extracellular/luminal epitope found on both APP and betaCTFs, gave more cell surface labeling of calpeptin-treated cells than control cells. Quantitative binding of this antibody confirmed that inhibiting calpains caused a partial redistribution of APP to the cell surface. These results demonstrate that 1) calpain inhibition results in a partial redistribution of APP to the cell surface, 2) this redistribution leads to an increase in both alpha- and beta-cleavage without changing the ratio of alphaCTFs/betaCTFs, and 3) the bulk of the betaCTFs in the cell are within early endosomes, confirming the importance of this compartment in APP processing. PMID- 12087105 TI - Crystal structure of the platelet glycoprotein Ib(alpha) N-terminal domain reveals an unmasking mechanism for receptor activation. AB - Glycoprotein Ib (GPIb) is a platelet receptor with a critical role in mediating the arrest of platelets at sites of vascular damage. GPIb binds to the A1 domain of von Willebrand factor (vWF-A1) at high blood shear, initiating platelet adhesion and contributing to the formation of a thrombus. To investigate the molecular basis of GPIb regulation and ligand binding, we have determined the structure of the N-terminal domain of the GPIb(alpha) chain (residues 1-279). This structure is the first determined from the cell adhesion/signaling class of leucine-rich repeat (LRR) proteins and reveals the topology of the characteristic disulfide-bonded flanking regions. The fold consists of an N-terminal beta hairpin, eight leucine-rich repeats, a disulfide-bonded loop, and a C-terminal anionic region. The structure also demonstrates a novel LRR motif in the form of an M-shaped arrangement of three tandem beta-turns. Negatively charged binding surfaces on the LRR concave face and anionic region indicate two-step binding kinetics to vWF-A1, which can be regulated by an unmasking mechanism involving conformational change of a key loop. Using molecular docking of the GPIb and vWF A1 crystal structures, we were also able to model the GPIb.vWF-A1 complex. PMID- 12087106 TI - The elevation of glutamate content and the amplification of insulin secretion in glucose-stimulated pancreatic islets are not causally related. AB - Glucose increases insulin secretion by raising cytoplasmic Ca(2+) ([Ca(2+)](i)) in beta-cells (triggering pathway) and augmenting the efficacy of Ca(2+) on exocytosis (amplifying pathway). It has been suggested that glutamate formed from alpha-ketoglutarate is a messenger of the amplifying pathway (Maechler, P., and Wollheim, C. B. (1999) Nature 402, 685-689). This hypothesis was tested with mouse islets depolarized with 30 mm KCl (+ diazoxide) or with a saturating concentration of sulfonylurea. Because [Ca(2+)](i) was elevated under these conditions, insulin secretion was stimulated already in 0 mm glucose. The amplification of secretion produced by glucose was accompanied by an increase in islet glutamate. However, glutamine (0.5-2 mm) markedly augmented islet glutamate without affecting insulin secretion, whereas glucose augmented secretion without influencing glutamate levels when these were elevated by glutamine. Allosteric activation of glutamate dehydrogenase by BCH (2-amino 2-norbornane carboxylic acid) lowered islet glutamate but increased insulin secretion. Similar insulin secretion thus occurred at very different cellular glutamate levels. Glutamine did not affect islet [Ca(2+)](i) and pH(i), whereas glucose and BCH slightly raised pH(i) and either slightly decreased (30 mm KCl) or increased (tolbutamide) [Ca(2+)](i). The general dissociation between changes in islet glutamate and insulin secretion refutes a role of beta-cell glutamate in the amplification of insulin secretion by glucose. PMID- 12087107 TI - Regulation of vasodilator-stimulated phosphoprotein phosphorylation and interaction with Abl by protein kinase A and cell adhesion. AB - Members of the vasodilator-stimulated phosphoprotein (VASP) family are important regulators of actin cytoskeletal dynamics whose functions and protein-protein interactions are regulated by phosphorylation by the cAMP-dependent protein kinase (PKA). Herein, we show that phosphorylation of VASP is dynamically regulated by cellular adhesion to extracellular matrix. Detachment of cells stimulated PKA activity and induced PKA-dependent phosphorylation of VASP and the related murine-Enabled (Mena) protein. VASP and Mena were rapidly dephosphorylated immediately following reattachment but showed an intermediate level of phosphorylation during active cell spreading. This pattern correlated closely with adhesion-dependent changes in PKA activity. The in vivo interaction of VASP with the Abl tyrosine kinase, shown here for the first time, was readily apparent in adherent cells, lost following cellular detachment, and induced upon reattachment to matrix. Importantly, inhibition of PKA activity prevented phosphorylation of VASP and dissociation of VASP-Abl complexes after cellular detachment, whereas activation of PKA completely eliminated the co immunoprecipitation of Abl activity with VASP. These data establish a new biochemical link between cell adhesion and regulation of VASP proteins and provide the first demonstration of a regulated interaction between VASP and Abl in mammalian cells. PMID- 12087108 TI - Plasmin-induced migration of endothelial cells. A potential target for the anti angiogenic action of angiostatin. AB - Angiostatin, a plasminogen fragment containing 3-4 N-terminal kringle domains, is a potent inhibitor of tumor-induced angiogenesis, but its mechanism of action is unclear. Angiostatin is a ligand for integrin alphavbeta(3) but does not induce stress fiber formation upon integrin binding, suggesting that angiostatin is a potential integrin antagonist. Plasmin, the parent molecule of angiostatin and a major extracellular protease, induces platelet aggregation, migration of peripheral blood monocytes, and release of arachidonate and leukotriene from several cell types. In the current study, we found that plasmin specifically bound to alphavbeta(3) through the kringle domains and induced migration of endothelial cells. In contrast, angiostatin did not induce cell migration. Notably, angiostatin, anti-alphavbeta(3) antibodies, RGD-peptide, and a serine protease inhibitor effectively blocked plasmin-induced cell migration. These results suggest that plasmin-induced migration of endothelial cells requires alphavbeta(3) and the catalytic activity of plasmin and that this process is a potential target for the inhibitory activity of angiostatin. PMID- 12087109 TI - The Saccharomyces cerevisiae YBR159w gene encodes the 3-ketoreductase of the microsomal fatty acid elongase. AB - The YBR159w gene encodes the major 3-ketoreductase activity of the elongase system of enzymes required for very long-chain fatty acid (VLCFA) synthesis. Mutants lacking the YBR159w gene display many of the phenotypes that have previously been described for mutants with defects in fatty acid elongation. These phenotypes include reduced VLCFA synthesis, accumulation of high levels of dihydrosphingosine and phytosphingosine, and accumulation of medium-chain ceramides. In vitro elongation assays confirm that the ybr159Delta mutant is deficient in the reduction of the 3-ketoacyl intermediates of fatty acid elongation. The ybr159Delta mutant also displays reduced dehydration of the 3-OH acyl intermediates of fatty acid elongation, suggesting that Ybr159p is required for the stability or function of the dehydratase activity of the elongase system. Green fluorescent protein-tagged Ybr159p co-localizes and co-immunoprecipitates with other elongating enzymes, Elo3p and Tsc13p. Whereas VLCFA synthesis is essential for viability, the ybr159Delta mutant cells are viable (albeit very slowly growing) and do synthesize some VLCFA. This suggested that a functional ortholog of Ybr159p exists that is responsible for the residual 3-ketoreductase activity. By disrupting the orthologs of Ybr159w in the ybr159Delta mutant we found that the ybr159Deltaayr1Delta double mutant was inviable, suggesting that Ayr1p is responsible for the residual 3-ketoreductase activity. PMID- 12087110 TI - Intracellular trafficking and membrane targeting mechanisms of the human reduced folate carrier in Mammalian epithelial cells. AB - The major pathway for cellular uptake of the water-soluble vitamin folic acid in mammalian cells is via a plasma membrane protein known as the reduced folate carrier (RFC). The molecular determinants that dictate plasma membrane expression of RFC as well as the cellular mechanisms that deliver RFC to the cell surface remain poorly defined. Therefore, we designed a series of fusion proteins of the human RFC (hRFC) with green fluorescent protein to image the targeting and trafficking dynamics of hRFC in living epithelial cells. We show that, in contrast to many other nutrient transporters, the molecular determinants that dictate hRFC plasma membrane expression reside within the hydrophobic backbone of the polypeptide and not within the cytoplasmic NH(2)- or COOH-terminal domains of the protein. Further, the integrity of the hRFC backbone is critical for export of the polypeptide from the endoplasmic reticulum to the cell surface. This trafficking is critically dependent on intact microtubules because microtubule disruption inhibits motility of hRFC-containing vesicles as well as final expression of hRFC in the plasma membrane. For the first time, these data define the mechanisms that control the intracellular trafficking and cell surface localization of hRFC within mammalian epithelia. PMID- 12087111 TI - The key role of anaplerosis and cataplerosis for citric acid cycle function. PMID- 12087112 TI - Investigating users' requirements: computer-based anatomy learning modules for multiple user test beds. AB - OBJECTIVE: User data and information about anatomy education were used to guide development of a learning environment that is efficient and effective. The research question focused on how to design instructional software suitable for the educational goals of different groups of users of the Visible Human data set. The ultimate goal of the study was to provide options for students and teachers to use different anatomy learning modules corresponding to key topics, for course work and professional training. DESIGN: The research used both qualitative and quantitative methods. It was driven by the belief that good instructional design must address learning context information and pedagogic content information. The data collection emphasized measurement of users' perspectives, experience, and demands in anatomy learning. MEASUREMENT: Users' requirements elicited from 12 focus groups were combined and rated by 11 researchers. Collective data were sorted and analyzed by use of multidimensional scaling and cluster analysis. RESULTS: A set of functions and features in high demand across all groups of users was suggested by the results. However, several subgroups of users shared distinct demands. The design of the learning modules will encompass both unified core components and user-specific applications. The design templates will allow sufficient flexibility for dynamic insertion of different learning applications for different users. CONCLUSION: This study describes how users' requirements, associated with users' learning experiences, were systematically collected and analyzed and then transformed into guidelines informing the iterative design of multiple learning modules. Information about learning challenges and processes was gathered to define essential anatomy teaching strategies. A prototype instrument to design and polish the Visible Human user interface system is currently being developed using ideas and feedback from users. PMID- 12087115 TI - "You have to be there": twenty-five years of SCAMC/AMIA symposia. AB - The AMIA Annual Symposium began life as the Symposium on Computer Applications in Medical Care (SCAMC) in 1977. Inaugurated as a multidisciplinary meeting with multiple sponsors, the Symposium quickly became a force in the development of the field of medical informatics. The authors summarize the 25-year history of the meeting and its proceedings, drawing on information in the printed programs and proceedings and on the personal recollections of some Symposium organizers and attendees. They also present the results of a study of the extent to which Symposium papers have been cited in the journal literature. PMID- 12087113 TI - A user-centered model for web site design: needs assessment, user interface design, and rapid prototyping. AB - As the Internet continues to grow as a delivery medium for health information, the design of effective Web sites becomes increasingly important. In this paper, the authors provide an overview of one effective model for Web site design, a user-centered process that includes techniques for needs assessment, goal/task analysis, user interface design, and rapid prototyping. They detail how this approach was employed to design a family health history Web site, Health Heritage . This Web site helps patients record and maintain their family health histories in a secure, confidential manner. It also supports primary care physicians through analysis of health histories, identification of potential risks, and provision of health care recommendations. Visual examples of the design process are provided to show how the use of this model resulted in an easy-to-use Web site that is likely to meet user needs. The model is effective across diverse content arenas and is appropriate for applications in varied media. PMID- 12087117 TI - Arden/J: an architecture for MLM execution on the Java platform. AB - The Arden Syntax was introduced more than 10 years ago, but it is still not in widespread use. One reason might be that for each particular architecture and information system, a different Arden Syntax compiler must be written as well as a program for the runtime execution of the medical logic modules (MLMs). The authors have designed and implemented an architecture that increases the portability of Arden Syntax rules, using the Java platform. The portability to a target information system is achieved by the addition of appropriate adapter components, which they call mappers. These mappers are dynamically selected using explicit and implicit elements of MLMs. Furthermore, they can help translate data from the clinical information system representation into the representation needed by an MLM. This was validated by an experiment in two clinical units. Also, the authors propose a convention to name signals that trigger other MLMs (called intermediate states) so that they remain unique to each institution. The authors implemented this architecture in their clinical system and in an XML based medical record application that has been used experimentally in their urology and nephrology departments. The Tetrasys company that provided the medical record was able to incorporate their runtime without modifications, and typical MLM execution time was less than 1 sec. PMID- 12087118 TI - Metadata-driven ad hoc query of patient data: meeting the needs of clinical studies. AB - Clinical study data management systems (CSDMSs) have many similarities to clinical patient record systems (CPRSs) in their focus on recording clinical parameters. Requirements for ad hoc query interfaces for both systems would therefore appear to be highly similar. However, a clinical study is concerned primarily with collective responses of groups of subjects to standardized therapeutic interventions for the same underlying clinical condition. The parameters that are recorded in CSDMSs tend to be more diverse than those required for patient management in non-research settings, because of the greater emphasis on questionnaires for which responses to each question are recorded separately. The differences between CSDMSs and CPRSs are reflected in the metadata that support the respective systems' operation, and need to be reflected in the query interfaces. The authors describe major revisions of their previously described CSDMS ad hoc query interface to meet CSDMS needs more fully, as well as its porting to a Web-based platform. PMID- 12087116 TI - Computer-generated patient education materials: do they affect professional practice? A systematic review. AB - A systematic search of seven electronic databases was done to identify randomized controlled trials that assessed the effect of computer-generated patient education material (PEM) on professional practice. Three studies met the authors' criteria. All three studies involved preventive care. All used a complex intervention of which computer-generated PEM was a major component. Improvements in practice were seen in all studies, although these gains were generally modest. One study showed improvement in patient outcomes. Mann-Whitney statistics calculated for the studies' outcome measures ranged from 0.48 to 0.66, equivalent to risk differences of -4 to 32 percent. Computer-generated PEM seems to have a small, positive effect on professional practice. The small number of included studies and the complex nature of the interventions makes it difficult to draw conclusions about the ability of computer-generated PEM to change professional practice. Future work should involve well-defined interventions that can be clearly evaluated in terms of effect and cost. PMID- 12087119 TI - Effects of a multimedia project on users' knowledge about normal forgetting and serious memory loss. AB - OBJECTIVE: The aim of the project was to develop and evaluate the effectiveness of a CD-ROM-based multimedia program as a tool to increase user's knowledge about the differences between "normal" forgetfulness and more serious memory loss associated with Alzheimer's disease. DESIGN AND MEASUREMENTS: The research was a controlled randomized study conducted with 113 adults who were recruited from the community and who expressed a concern about memory loss in a family member. The intervention group (n=56) viewed a module entitled "Forgetfulness: What's Normal and What's Not" on a laptop computer in their homes; the control group (n=57) did not. Both groups completed a 25-item knowledge-about-memory-loss test (primary outcome) and a sociodemographic and technology usage questionnaire; the intervention group also completed a CD-ROM user's evaluation. RESULTS: The mean (SD) number of correct responses to the knowledge test was 14.2 (4.5) for controls and 19.7 (3.1) for intervention participants. This highly significant difference (p<0.001) corresponds to a very large effect size. The program was most effective for participants with a lower level of self-reported prior knowledge about memory loss and Alzheimer's disease (p=0.02). Viewers were very satisfied with the program and felt that it was easy to use and understand. They particularly valued having personal access to a confidential source that permitted them to become informed about memory loss without public disclosure. CONCLUSION: This multimedia CD-ROM technology program provides an efficient and effective means of teaching older adults about memory loss and ways to distinguish benign from serious memory loss. It uniquely balances public community outreach education and personal privacy. PMID- 12087120 TI - Does feedback improve the quality of computerized medical records in primary care? AB - OBJECTIVE: The MediPlus database collects anonymized information from generalpractice computer systems in the United Kingdom, for research purposes. Data quality markers are collated and fed back to the participating general practitioners. The authors examined whether this feedback had a significant effect on data quality. METHODS: The data quality markers used since 1992 were examined. The authors determined whether the feedback of "useful" data quality markers led to a statistically significant improvement in these markers. Environmental influences on data quality from outside the scheme were controlled for by examination of the data quality scores of new entrants. RESULTS: Three quality markers improved significantly over the period of the study. These were the use of highly specific "lower-level" Read Codes (p=0.004) and the linkage of repeat prescriptions (p=0.03) and acute prescriptions (p=0.04) to diagnosis. Clinicians who fall below the target level for linkage of repeat prescriptions to diagnosis receive more detailed feedback; the effect of this was also statistically significant (p<0.01.) CONCLUSIONS: The feedback of four of the ten markers had a significant effect on data quality. The effect of more detailed feedback appears to have had a greater effect. The lessons learned from this approach may help improve the quality of electronic medical records in the United Kingdom and elsewhere. PMID- 12087121 TI - Consumer health information on the Internet. PMID- 12087123 TI - Presentation of Morris F. Collen Award to professors Howard Bleich and Warner Slack. PMID- 12087126 TI - Distinct domains of splicing factor Prp8 mediate different aspects of spliceosome activation. AB - Prp8 is the largest and most highly conserved protein in the spliceosome yet its mechanism of function is poorly understood. Our previous studies implicate Prp8 in control of spliceosome activation for the first catalytic step of splicing, because substitutions in five distinct regions (a-e) of Prp8 suppress a cold sensitive block to activation caused by a mutation in U4 RNA. Catalytic activation of the spliceosome is thought to require unwinding of the U1 RNA/5' splice site and U4/U6 RNA helices by the Prp28 and Prp44/Brr2 DExD/H-box helicases, respectively. Here we show that mutations in regions a, d, and e of Prp8 exhibit allele-specific genetic interactions with mutations in Prp28, Prp44/Brr2, and U6 RNA, respectively. These results indicate that Prp8 coordinates multiple processes in spliceosome activation and enable an initial correlation of Prp8 structure and function. Furthermore, additional genetic interactions with U4-cs1 support a two-state model for this RNA conformational switch and implicate another splicing factor, Prp31, in Prp8-mediated spliceosome activation. PMID- 12087127 TI - GATA-3 transcriptional imprinting in Th2 lymphocytes: a mathematical model. AB - Immunological memory involves the fast recall of cytokine expression by T helper (Th) lymphocytes. Two distinct profiles of cytokine expression, Th1 and Th2, can be induced by antigen and polarizing signals during activation of naive Th cells and can subsequently be reexpressed on stimulation by antigen alone. The transcription factor GATA-3 induces Th2 development. GATA-3 is activated by the Th2-polarizing stimulus, IL-4, and has recently been observed to autoactivate its transcription. Based on these experimental data, we developed a mathematical model of GATA-3 expression that assumes independent activation of GATA-3 transcription by IL-4 and by GATA-3. Cooperativity of GATA-3 transcriptional activation is shown to create a threshold for autoactivation, resulting in the coexistence of two distinct GATA-3 expression states: a state of basal expression and a state of high expression sustained by autoactivation. Suprathreshold IL-4 signals induce a transition from basal to high GATA-3 expression. Thus, GATA-3 autoactivation creates a bistable system that can memorize a transient inductive signal. The model further predicts conditions under which the state of high GATA 3 expression can be abolished, which may extinguish the Th2 cytokine memory. PMID- 12087128 TI - Role of the short isoform of myosin light chain kinase in the contraction of cultured smooth muscle cells as examined by its down-regulation. AB - GbaSM-4 cells, smooth muscle cells derived from brain basilar artery, which express both 210-kDa long and 130-kDa short isoforms of myosin light chain kinase (MLCK), were infected with an adenovirus vector carrying a 1.4-kb catalytic portion of MLCK-cDNA in an antisense orientation. Western blot analysis showed that the expression of short MLCK was depressed without affecting long MLCK expression. The contraction of the down-regulated cells was measured by the cell populated collagen-fiber method. The tension development after stimulation with norepinephrine or was depressed. The additional infection of the down-regulated cells with the adenovirus construct containing the same insert in a sense direction rescued not only the short MLCK expression but also contraction, confirming the physiological role of short MLCK in the contraction. To examine the role of long MLCK in the residual contraction persisting in the short MLCK deficient cells, long MLCK was further down-regulated by increasing the multiplicity of infection of the antisense construct. The additional down regulation of long MLCK expression, however, did not alter the residual contraction, ruling out the involvement of long MLCK in the contractile activity. Further, in the cells where short MLCK was down-regulated specifically, the extent of phosphorylation of 20-kDa myosin light chain (MLC20) after the agonist stimulation was not affected. This finding suggests that there are additional factors to MLC20 phosphorylation that contribute to regulate smooth muscle contraction. PMID- 12087129 TI - Lung epithelial fluid transport and the resolution of pulmonary edema. AB - The discovery of mechanisms that regulate salt and water transport by the alveolar and distal airway epithelium of the lung has generated new insights into the regulation of lung fluid balance under both normal and pathological conditions. There is convincing evidence that active sodium and chloride transporters are expressed in the distal lung epithelium and are responsible for the ability of the lung to remove alveolar fluid at the time of birth as well as in the mature lung when pathological conditions lead to the development of pulmonary edema. Currently, the best described molecular transporters are the epithelial sodium channel, the cystic fibrosis transmembrane conductance regulator, Na+-K+-ATPase, and several aquaporin water channels. Both catecholamine-dependent and -independent mechanisms can upregulate isosmolar fluid transport across the distal lung epithelium. Experimental and clinical studies have made it possible to examine the role of these transporters in the resolution of pulmonary edema. PMID- 12087130 TI - Plastic changes in the central auditory system after hearing loss, restoration of function, and during learning. AB - Traditionally the auditory system was considered a hard-wired sensory system; this view has been challenged in recent years in light of the plasticity of other sensory systems, particularly the visual and somatosensory systems. Practical experience in clinical audiology together with the use of prosthetic devices, such as cochlear implants, contributed significantly to the present view on the plasticity of the central auditory system, which was originally based on data obtained in animal experiments. The loss of auditory receptors, the hair cells, results in profound changes in the structure and function of the central auditory system, typically demonstrated by a reorganization of the projection maps in the auditory cortex. These plastic changes occur not only as a consequence of mechanical lesions of the cochlea or biochemical lesions of the hair cells by ototoxic drugs, but also as a consequence of the loss of hair cells in connection with aging or noise exposure. In light of the aging world population and the increasing amount of noise in the modern world, understanding the plasticity of the central auditory system has its practical consequences and urgency. In most of these situations, a common denominator of central plastic changes is a deterioration of inhibition in the subcortical auditory nuclei and the auditory cortex. In addition to the processes that are elicited by decreased or lost receptor function, the function of nerve cells in the adult central auditory system may dynamically change in the process of learning. A better understanding of the plastic changes in the central auditory system after sensory deafferentation, sensory stimulation, and learning may contribute significantly to improvement in the rehabilitation of damaged or lost auditory function and consequently to improved speech processing and production. PMID- 12087131 TI - Modification of brain aging and neurodegenerative disorders by genes, diet, and behavior. AB - Multiple molecular, cellular, structural, and functional changes occur in the brain during aging. Neural cells may respond to these changes adaptively, or they may succumb to neurodegenerative cascades that result in disorders such as Alzheimer's and Parkinson's diseases. Multiple mechanisms are employed to maintain the integrity of nerve cell circuits and to facilitate responses to environmental demands and promote recovery of function after injury. The mechanisms include production of neurotrophic factors and cytokines, expression of various cell survival-promoting proteins (e.g., protein chaperones, antioxidant enzymes, Bcl-2 and inhibitor of apoptosis proteins), preservation of genomic integrity by telomerase and DNA repair proteins, and mobilization of neural stem cells to replace damaged neurons and glia. The aging process challenges such neuroprotective and neurorestorative mechanisms. Genetic and environmental factors superimposed upon the aging process can determine whether brain aging is successful or unsuccessful. Mutations in genes that cause inherited forms of Alzheimer's disease (amyloid precursor protein and presenilins), Parkinson's disease (alpha-synuclein and Parkin), and trinucleotide repeat disorders (huntingtin, androgen receptor, ataxin, and others) overwhelm endogenous neuroprotective mechanisms; other genes, such as those encoding apolipoprotein E(4), have more subtle effects on brain aging. On the other hand, neuroprotective mechanisms can be bolstered by dietary (caloric restriction and folate and antioxidant supplementation) and behavioral (intellectual and physical activities) modifications. At the cellular and molecular levels, successful brain aging can be facilitated by activating a hormesis response in which neurons increase production of neurotrophic factors and stress proteins. Neural stem cells that reside in the adult brain are also responsive to environmental demands and appear capable of replacing lost or dysfunctional neurons and glial cells, perhaps even in the aging brain. The recent application of modern methods of molecular and cellular biology to the problem of brain aging is revealing a remarkable capacity within brain cells for adaptation to aging and resistance to disease. PMID- 12087132 TI - Vascular growth factors and lymphangiogenesis. AB - Blood and lymphatic vessels develop in a parallel, but independent manner, and together form the circulatory system allowing the passage of fluid and delivering molecules within the body. Although the lymphatic vessels were discovered already 300 years ago, at the same time as the blood circulation was described, the lymphatic system has remained relatively neglected until recently. This is in part due to the difficulties in recognizing these vessels in tissues because of a lack of specific markers. Over the past few years, several molecules expressed specifically in the lymphatic endothelial cells have been characterized, and knowledge about the lymphatic system has started to accumulate again. The vascular endothelial growth factor (VEGF) family of growth factors and receptors is involved in the development and growth of the vascular endothelial system. Two of its family members, VEGF-C and VEGF-D, regulate the lymphatic endothelial cells via their receptor VEGFR-3. With the aid of these molecules, lymphatic endothelial cells can be isolated and cultured, allowing detailed studies of the molecular properties of these cells. Also the role of the lymphatic endothelium in immune responses and certain pathological conditions can be studied in more detail, as the blood and lymphatic vessels seem to be involved in many diseases in a coordinated manner. Discoveries made so far will be helpful in the diagnosis of certain vascular tumors, in the design of specific treatments for lymphedema, and in the prevention of metastatic tumor spread via the lymphatic system. PMID- 12087133 TI - Polarized calcium and calmodulin signaling in secretory epithelia. AB - This review examines polarized calcium and calmodulin signaling in exocrine epithelial cells. The calcium ion is a simple, evolutionarily ancient, and universal second messenger. In exocrine epithelial cells, it regulates essential functions such as exocytosis, fluid secretion, and gene expression. Exocrine cells are structurally polarized, with the apical region usually dedicated to secretion. Recent advances in technology, in particular the development of videoimaging and confocal microscopy, have led to the discovery of polarized, subcellular calcium signals in these cell types. The properties of a rich variety of local and global calcium signals have now been described in secretory epithelial cells. Secretagogues stimulate apical-to-basal waves of calcium in many exocrine cell types, but there are some interesting exceptions to this rule. The shapes of intracellular calcium signals are determined by the distribution of calcium-releasing channels and mechanisms that limit calcium elevation. Polarized distribution of calcium-handling mechanisms also leads to transcellular calcium transport in exocrine epithelial cells. This transport can deliver considerable amounts of calcium into secreted fluids. Multicellular polarized calcium signals can coordinate the activity of many individual cells in epithelial secretory tissue. Certain particularly sensitive cells serve as pacemakers for initiation of intercellular calcium waves. Many calcium signaling pathways involve activation of calmodulin. This ubiquitous protein regulates secretion in exocrine cells and also activates interesting feedback interactions with calcium channels and transporters. Very recently it became possible to directly study polarized calcium-calmodulin reactions and to visualize the process of hormone-induced redistribution of calmodulin in live cells. The structural and functional polarity of secretory epithelia alongside the polarity of its calcium and calmodulin signaling present an interesting lesson in tissue organization. PMID- 12087134 TI - Epithelial sodium channel/degenerin family of ion channels: a variety of functions for a shared structure. AB - The recently discovered epithelial sodium channel (ENaC)/degenerin (DEG) gene family encodes sodium channels involved in various cell functions in metazoans. Subfamilies found in invertebrates or mammals are functionally distinct. The degenerins in Caenorhabditis elegans participate in mechanotransduction in neuronal cells, FaNaC in snails is a ligand-gated channel activated by neuropeptides, and the Drosophila subfamily is expressed in gonads and neurons. In mammals, ENaC mediates Na+ transport in epithelia and is essential for sodium homeostasis. The ASIC genes encode proton-gated cation channels in both the central and peripheral nervous system that could be involved in pain transduction. This review summarizes the physiological roles of the different channels belonging to this family, their biophysical and pharmacological characteristics, and the emerging knowledge of their molecular structure. Although functionally different, the ENaC/DEG family members share functional domains that are involved in the control of channel activity and in the formation of the pore. The functional heterogeneity among the members of the ENaC/DEG channel family provides a unique opportunity to address the molecular basis of basic channel functions such as activation by ligands, mechanotransduction, ionic selectivity, or block by pharmacological ligands. PMID- 12087135 TI - Cyclic nucleotide-gated ion channels. AB - Cyclic nucleotide-gated (CNG) channels are nonselective cation channels first identified in retinal photoreceptors and olfactory sensory neurons (OSNs). They are opened by the direct binding of cyclic nucleotides, cAMP and cGMP. Although their activity shows very little voltage dependence, CNG channels belong to the superfamily of voltage-gated ion channels. Like their cousins the voltage-gated K+ channels, CNG channels form heterotetrameric complexes consisting of two or three different types of subunits. Six different genes encoding CNG channels, four A subunits (A1 to A4) and two B subunits (B1 and B3), give rise to three different channels in rod and cone photoreceptors and in OSNs. Important functional features of these channels, i.e., ligand sensitivity and selectivity, ion permeation, and gating, are determined by the subunit composition of the respective channel complex. The function of CNG channels has been firmly established in retinal photoreceptors and in OSNs. Studies on their presence in other sensory and nonsensory cells have produced mixed results, and their purported roles in neuronal pathfinding or synaptic plasticity are not as well understood as their role in sensory neurons. Similarly, the function of invertebrate homologs found in Caenorhabditis elegans, Drosophila, and Limulus is largely unknown, except for two subunits of C. elegans that play a role in chemosensation. CNG channels are nonselective cation channels that do not discriminate well between alkali ions and even pass divalent cations, in particular Ca2+. Ca2+ entry through CNG channels is important for both excitation and adaptation of sensory cells. CNG channel activity is modulated by Ca2+/calmodulin and by phosphorylation. Other factors may also be involved in channel regulation. Mutations in CNG channel genes give rise to retinal degeneration and color blindness. In particular, mutations in the A and B subunits of the CNG channel expressed in human cones cause various forms of complete and incomplete achromatopsia. PMID- 12087136 TI - Increasing choices for HIV therapy. PMID- 12087137 TI - Oral contraceptives and the risk of breast cancer. AB - BACKGROUND: It is uncertain whether the use of an oral contraceptive increases the risk of breast cancer later in life, when the incidence of breast cancer is increased. We conducted a population-based, case-control study to determine the risk of breast cancer among former and current users of oral contraceptives. METHODS: We interviewed women who were 35 to 64 years old. A total of 4575 women with breast cancer and 4682 controls were interviewed. Conditional logistic regression was used to calculate odds ratios as estimates of the relative risk (incidence-density ratios) of breast cancer. RESULTS: The relative risk was 1.0 (95 percent confidence interval, 0.8 to 1.3) for women who were currently using oral contraceptives and 0.9 (95 percent confidence interval, 0.8 to 1.0) for those who had previously used them. The relative risk did not increase consistently with longer periods of use or with higher doses of estrogen. The results were similar among white and black women. Use of oral contraceptives by women with a family history of breast cancer was not associated with an increased risk of breast cancer, nor was the initiation of oral-contraceptive use at a young age. CONCLUSIONS: Among women from 35 to 64 years of age, current or former oral-contraceptive use was not associated with a significantly increased risk of breast cancer. PMID- 12087138 TI - Lansoprazole for the prevention of recurrences of ulcer complications from long term low-dose aspirin use. AB - BACKGROUND: The role of gastric acid suppression in preventing the recurrence of ulcer complications after the eradication of Helicobacter pylori infection in patients taking long-term low-dose aspirin is uncertain. METHODS: We enrolled 123 patients who had ulcer complications after using low-dose aspirin continuously for more than one month and who had H. pylori infection. After the ulcers had healed and the H. pylori infection was eradicated, the patients were randomly assigned to treatment with 30 mg of lansoprazole daily or placebo, in addition to 100 mg of aspirin daily, for 12 months. The primary end point was the recurrence of ulcer complications. RESULTS: During a median follow-up of 12 months, 9 of the 61 patients in the placebo group (14.8 percent), as compared with 1 of the 62 patients in the lansoprazole group (1.6 percent), had a recurrence of ulcer complications (adjusted hazard ratio, 9.6; 95 percent confidence interval, 1.2 to 76.1). Of these 10 patients, 4 had evidence of a recurrence of H. pylori infection and 2 had taken nonsteroidal antiinflammatory drugs before the onset of complications. Patients in the lansoprazole group were significantly less likely to have a recurrence of ulcer complications than patients in the placebo group (P=0.008). There was no significant difference in mortality between the two groups. CONCLUSIONS: In patients who had ulcer complications related to the long term use of low-dose aspirin, treatment with lansoprazole in addition to the eradication of H. pylori infection significantly reduced the rate of recurrence of ulcer complications. PMID- 12087139 TI - Lopinavir-ritonavir versus nelfinavir for the initial treatment of HIV infection. AB - BACKGROUND: Lopinavir is a newly developed inhibitor of human immunodeficiency virus (HIV) protease that, when formulated with ritonavir, yields mean trough plasma lopinavir concentrations that are at least 75 times as high as that needed to inhibit replication of wild-type HIV by 50 percent. METHODS: We conducted a double-blind trial in which 653 HIV-infected adults who had not received antiretroviral therapy for more than 14 days were randomly assigned to receive either lopinavir-ritonavir (400 mg of lopinavir plus 100 mg of ritonavir twice daily) with nelfinavir placebo or nelfinavir (750 mg three times daily) with lopinavir-ritonavir placebo. All patients also received open-label stavudine and lamivudine. The primary efficacy end points were the presence of fewer than 400 HIV RNA copies per milliliter of plasma at week 24 and the time to the loss of virologic response through week 48. RESULTS: At week 48, greater proportions of patients treated with lopinavir-ritonavir than of patients treated with nelfinavir had fewer than 400 copies of HIV RNA per milliliter (75 percent vs. 63 percent, P<0.001) and fewer than 50 copies per milliliter (67 percent vs. 52 percent, P<0.001). The time to the loss of virologic response was greater in the lopinavir-ritonavir group than in the nelfinavir group (hazard ratio, 2.0; 95 percent confidence interval, 1.5 to 2.7; P<0.001). The estimated proportion of patients with a persistent virologic response through week 48 was 84 percent for patients receiving lopinavir-ritonavir and 66 percent for those receiving nelfinavir. Both regimens were well tolerated, with the rate of discontinuation related to the study drugs at 3.4 percent among patients receiving lopinavir ritonavir and 3.7 percent among patients receiving nelfinavir. Among patients with more than 400 copies of HIV RNA per milliliter at some point from week 24 through week 48, resistance mutations in HIV protease were demonstrated in viral isolates from 25 of 76 nelfinavir-treated patients (33 percent) and none of 37 patients treated with lopinavir-ritonavir (P<0.001). CONCLUSIONS: For the initial treatment of HIV-infected adults, a combination regimen that includes lopinavir ritonavir is well tolerated and has antiviral activity superior to that of a nelfinavir-containing regimen. PMID- 12087140 TI - Troponin T levels in patients with acute coronary syndromes, with or without renal dysfunction. AB - BACKGROUND: Among patients with suspected acute coronary syndromes, cardiac troponin T levels have prognostic value. However, there is concern that renal dysfunction may impair the prognostic value, because cardiac troponin T may be cleared by the kidney. METHODS: We analyzed the outcomes in 7033 patients enrolled in the Global Use of Strategies to Open Occluded Coronary Arteries IV trial who had complete base-line data on troponin T levels and creatinine clearance rates. The troponin T level was considered abnormal if it was 0.1 ng per milliliter or higher, and creatinine clearance was assessed in quartiles. The primary end point was a composite of death or myocardial infarction within 30 days. RESULTS: Death or myocardial infarction occurred in 581 patients. Among patients with a creatinine clearance above the 25th percentile value of 58.4 ml per minute, an abnormally elevated troponin T level was predictive of an increased risk of myocardial infarction or death (7 percent vs. 5 percent; adjusted odds ratio, 1.7; 95 percent confidence interval, 1.3 to 2.2; P<0.001). Among patients with a creatinine clearance in the lowest quartile, an elevated troponin T level was similarly predictive of increased risk (20 percent vs. 9 percent; adjusted odds ratio, 2.5; 95 percent confidence interval, 1.8 to 3.3; P<0.001). When the creatinine clearance rate was considered as a continuous variable and age, sex, ST-segment depression, heart failure, previous revascularization, diabetes mellitus, and other confounders had been accounted for, elevation of the troponin T level was independently predictive of risk across the entire spectrum of renal function. CONCLUSIONS: Cardiac troponin T levels predict short-term prognosis in patients with acute coronary syndromes regardless of their level of creatinine clearance. PMID- 12087141 TI - Antenatal membranous glomerulonephritis due to anti-neutral endopeptidase antibodies. PMID- 12087142 TI - Image in clinical medicine. Disseminated histoplasmosis in a patient with human immunodeficiency virus infection. PMID- 12087143 TI - Implantable devices for the treatment of atrial fibrillation. PMID- 12087144 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 20-2002. A 37-year-old man with fever, hepatosplenomegaly, and a cutaneous foot lesion after a trip to Africa. PMID- 12087145 TI - Good news about oral contraceptives. PMID- 12087146 TI - Decision making with cardiac troponin tests. PMID- 12087147 TI - The genetic gymnastics of our indigenous microbes. PMID- 12087148 TI - Monoclonal gammopathy of undetermined significance. PMID- 12087149 TI - Bisphosphonates and osteoporosis. PMID- 12087150 TI - Strategies to improve outcomes after renal transplantation. PMID- 12087151 TI - Pharmacotherapy for obesity. PMID- 12087152 TI - Protecting research subjects. PMID- 12087153 TI - Seizure associated with the use of visicol for colonoscopy. PMID- 12087155 TI - Opposing effects of Kruppel-like factor 4 (gut-enriched Kruppel-like factor) and Kruppel-like factor 5 (intestinal-enriched Kruppel-like factor) on the promoter of the Kruppel-like factor 4 gene. AB - KLF4 (Kruppel-like factor 4 or gut-enriched Kruppel-like factor, GKLF) and KLF5 (Kruppel-like factor 5 or intestinal-enriched Kruppel-like factor, IKLF) are two closely related members of the zinc finger-containing Kruppel-like factor family of transcription factors. Although both genes are expressed in the intestinal epithelium, their distributions are different: Klf4 is primarily expressed in the terminally differentiated villus cells while Klf5 is primarily in the proliferating crypt cells. Previous studies show that Klf4 is a negative regulator of cell proliferation and Klf5 is a positive regulator of cell proliferation. In this study, we demonstrate that Klf5 binds to a number of cis DNA elements that have previously been shown to bind to Klf4. However, while Klf4 activates the promoter of its own gene, Klf5 suppresses the Klf4 promoter. Moreover, Klf5 abrogates the activating effect of Klf4 on the Klf4 promoter and Klf4 abrogates the inhibitory effect of Klf5 on the same promoter. An explanation of this competing effect is due to physical competition of the two proteins for binding to cognate DNA sequence. The complementary tissue localization of expression of Klf4 and Klf5 and the opposing effect of the two Klfs on the Klf4 promoter activity may provide a basis for the coordinated regulation of expression of the Klf4 gene in the intestinal epithelium. PMID- 12087154 TI - Spontaneous and double-strand break-induced recombination, and gene conversion tract lengths, are differentially affected by overexpression of wild-type or ATPase-defective yeast Rad54. AB - Rad54 plays key roles in homologous recombination (HR) and double-strand break (DSB) repair in yeast, along with Rad51, Rad52, Rad55 and Rad57. Rad54 belongs to the Swi2/Snf2 family of DNA-stimulated ATPases. Rad51 nucleoprotein filaments catalyze DNA strand exchange and Rad54 augments this activity of Rad51. Mutations in the Rad54 ATPase domain (ATPase(-)) impair Rad54 function in vitro, sensitize yeast to killing by methylmethane sulfonate and reduce spontaneous gene conversion. We found that overexpression of ATPase(-) Rad54 reduced spontaneous direct repeat gene conversion and increased both spontaneous direct repeat deletion and spontaneous allelic conversion. Overexpression of ATPase(-) Rad54 decreased DSB-induced allelic conversion, but increased chromosome loss and DSB dependent lethality. Thus, ATP hydrolysis by Rad54 contributes to genome stability by promoting high-fidelity DSB repair and suppressing spontaneous deletions. Overexpression of wild-type Rad54 did not alter DSB-induced HR levels, but conversion tract lengths were reduced. Interestingly, ATPase(-) Rad54 decreased overall HR levels and increased tract lengths. These tract length changes provide new in vivo evidence that Rad54 functions in the post-synaptic phase during recombinational repair of DSBs. PMID- 12087157 TI - Antisense properties of tricyclo-DNA. AB - Tricyclo (tc)-DNA belongs to the class of conformationally constrained DNA analogs that show enhanced binding properties to DNA and RNA. We prepared tc oligonucleotides up to 17 nt in length, and evaluated their binding efficiency and selectivity towards complementary RNA, their biological stability in serum, their RNase H inducing potential and their antisense activity in a cellular assay. Relative to RNA or 2'-O-Me-phosphorothioate (PS)-RNA, fully modified tc oligodeoxynucleotides, 10-17 nt in length, show enhanced selectivity and enhanced thermal stability by approximately 1 degrees C/modification in binding to RNA targets. Tricyclodeoxyoligonucleotides are completely stable in heat-deactivated fetal calf serum at 37 degree C. Moreover, tc-DNA-RNA duplexes are not substrates for RNase H. To test for antisense effects in vivo, we used HeLa cell lines stably expressing the human beta-globin gene with two different point mutations in the second intron. These mutations lead to the inclusion of an aberrant exon in beta-globin mRNA. Lipofectamine-mediated delivery of a 17mer tc oligodeoxynucleotide complementary to the 3'-cryptic splice site results in correction of aberrant splicing already at nanomolar concentrations with up to 100-fold enhanced efficiency relative to a 2'-O-Me-PS-RNA oligonucleotide of the same length and sequence. In contrast to 2'-O-Me-PS-RNA, tc-DNA shows antisense activity even in the absence of lipofectamine, albeit only at much higher oligonucleotide concentrations. PMID- 12087156 TI - Rad51p and Rad54p, but not Rad52p, elevate gene repair in Saccharomyces cerevisiae directed by modified single-stranded oligonucleotide vectors. AB - Synthetic single-stranded DNA vectors have been used to correct point and frameshift mutations in episomal or chromosomal targets in the yeast Saccharomyces cerevisiae. Certain parameters, such as the length of the vector and the genetic background of the organism, have a significant impact on the process of targeted gene repair, and point mutations are corrected at a higher frequency than frameshift mutations. Genetic analyses reveal that expression levels of the recombination/repair genes RAD51, RAD52 and RAD54 can affect the frequency of gene repair. Overexpression of RAD51 enhances the frequency 4-fold for correction of an episomal target and 5-fold for correction of a chromosomal target; overexpression of RAD54 is also effective in stimulating gene repair, to the same extent as RAD51 in the chromosomal target. In sharp contrast, RAD52 gene expression serves to reduce gene repair activity in rescue experiments and in experiments where RAD52 is overexpressed in a wild-type strain. This may suggest an antagonist role for Rad52p. Consistent with this notion, the highest level of targeted repair occurs when the RAD51 gene is overexpressed in a strain of yeast deficient in RAD52 gene function. PMID- 12087158 TI - Deficiency of a novel mismatch repair activity in a bladder tumor cell line. AB - We demonstrate here that a cell line derived from a bladder cancer is defective in strand-specific mismatch repair. The mismatch repair deficiency in this cell line is associated with microsatellite instability and blocks an early step in the repair pathway. Since the addition of a known mismatch repair component hMutSalpha, hMutSbeta, hMutLalpha, replication protein A or proliferating cellular nuclear antigen could not restore mismatch repair to the mutant extract, the bladder tumor cell line is likely to be defective in an uncharacterized repair component. However, the repair in the mutant extract could be complemented by a partially purified activity derived from HeLa nuclear extracts. Therefore, in addition to revealing that a loss of mismatch repair function is associated with bladder cancer, this study provides information implicating a new mismatch repair activity. PMID- 12087159 TI - Non-contact positions impose site selectivity on Cre recombinase. AB - A first step in Cre-mediated site-specific DNA recombination is binding to the two 13 bp repeats of the 34 bp site loxP. Several nucleotides within loxP do not directly contact the bound enzyme, yet mutation at two of these base pairs, at positions 11 and 12 in each repeat, results in a 100 000-fold reduction in recombination. To understand better how Cre selects DNA sequences for recombination, we combined DNA shuffling mutagenesis and a forward selection strategy to obtain Cre mutants that recombine at 100% efficiency a mutant loxK2 site carrying these dinucleotide changes. The role of the several mutations found in these Cre isolates was analyzed both in vivo and biochemically with purified enzymes. A single mutation at E262 accounts for most but not all of the enhanced activity at loxK2. Secondary mutations act in one or more of three ways: enhancement of loxK2 binding, accelerated synthesis of Cre in vivo or faster DNA recombination at the alternative spacer region present in loxK2. Systematic analysis of all 20 natural amino acids at position E262 shows that the naturally occurring glutamate residue at this position provides the optimal balance of efficiency of recombination at loxP and maximal discrimination against loxK2. PMID- 12087161 TI - DNA assembly using bis-peptide nucleic acids (bisPNAs). AB - DNA nanostructures are ordered oligonucleotide arrangements that have applications for DNA computers, crystallography, diagnostics and material sciences. Peptide nucleic acid (PNA) is a DNA/RNA mimic that offers many advantages for hybridization, but its potential for application in the field of DNA nanotechnology has yet to be thoroughly examined. We report the synthesis and characterization of tethered PNA molecules (bisPNAs) designed to assemble two individual DNA molecules through Watson-Crick base pairing. The spacer regions linking the PNAs were varied in length and contained amino acids with different electrostatic properties. We observed that bisPNAs effectively assembled oligonucleotides that were either the exact length of the PNA or that contained overhanging regions that projected outwards. In contrast, DNA assembly was much less efficient if the oligonucleotides contained overhanging regions that projected inwards. Surprisingly, the length of the spacer region between the PNA sequences did not greatly affect the efficiency of DNA assembly. Reasons for inefficient assembly of inward projecting DNA oligonucleotides include non sequence-specific intramolecular interactions between the overhanging region of the bisPNA and steric conflicts that complicate simultaneous binding of two inward projecting strands. These results suggest that bisPNA molecules can be used for self-assembling DNA nanostructures provided that the arrangement of the hybridizing DNA oligonucleotides does not interfere with simultaneous hybridization to the bisPNA molecule. PMID- 12087160 TI - Identification and characterization of transcription factor IIIA from Schizosaccharomyces pombe. AB - Transcription factor IIIA (TFIIIA) is specifically required for transcription of 5S rRNA genes and is the archetypal C2H2 zinc finger protein. All known vertebrate TFIIIAs have a similar organization: nine zinc fingers, followed by a C-terminal domain of unknown structure. The zinc fingers of Saccharomyces cerevisiae TFIIIA are interrupted between fingers eight and nine by an 81-amino acid spacer. Aside from the amino acids required for zinc finger folding, TFIIIAs from different species are remarkably divergent, whereas the natural binding site, the internal control region of the 5S rRNA gene, is well conserved. We now describe the identification and characterization of TFIIIA from Schizosaccharomyces pombe. This protein is organized differently from its known homologs, in that it contains eight closely spaced zinc fingers, a ninth zinc finger missing a C-terminal Zn2+-coordinating histidine, a 53- amino acid spacer, and an unprecedented tenth zinc finger. We have confirmed the identity of this divergent protein as TFIIIA by showing that it binds specifically and with high affinity to the S.pombe 5S rRNA gene. Comparison of DNase I protection patterns produced by TFIIIA from multiple species suggests a novel mode of DNA recognition by the S.pombe protein. Recombinant S.pombe TFIIIA was also shown to support specific transcription of the 5S rRNA gene in vitro. PMID- 12087162 TI - Use of fluorescent sequence-specific polyamides to discriminate human chromosomes by microscopy and flow cytometry. AB - In this paper, we demonstrate the use of synthetic polyamide probes to fluorescently label heterochromatic regions on human chromosomes for discrimination in cytogenetic preparations and by flow cytometry. Polyamides bind to the minor groove of DNA in a sequence-specific manner. Unlike conventional sequence-specific DNA or RNA probes, polyamides can recognize their target sequence without the need to subject chromosomes to harsh denaturing conditions. For this study, we designed and synthesized a polyamide to target the TTCCA-motif repeated in the heterochromatic regions of chromosome 9, Y and 1. We demonstrate that the fluorescently labeled polyamide binds to its target sequence in both conventional cytogenetic preparations of metaphase chromosomes and suspended chromosomes without denaturation. Chromosomes 9 and Y can be discriminated and purified by flow sorting on the basis of polyamide binding and Hoechst 33258 staining. We generate chromosome 9- and Y-specific 'paints' from the sorted fractions. We demonstrate the utility of this technology by characterizing the sequence of an olfactory receptor gene that is duplicated on multiple chromosomes. By separating chromosome 9 from chromosomes 10-12 on the basis of polyamide fluorescence, we determine and differentiate the haplotypes of the highly similar copies of this gene on chromosomes 9 and 11. PMID- 12087163 TI - High efficiency detection of bi-stranded abasic clusters in gamma-irradiated DNA by putrescine. AB - Bi-stranded abasic clusters, an abasic (AP) site on one DNA strand and another nearby AP site or strand break on the other, have been quantified using Nfo protein from Escherichia coli to produce a double-strand break at cluster sites. Since recent data suggest that Nfo protein cleaves inefficiently at some clusters, we tested whether polyamines, which also cut at AP sites, would cleave abasic clusters at higher efficiency. The data show that Nfo protein cleaves poorly at clusters containing immediately opposed AP sites and those separated by 1 or 3 bp. Putrescine (PUTR) cleaved more efficiently than spermidine or spermine, and did not cleave undamaged DNA. It cleaved abasic clusters in oligonucleotide duplexes more effectively than Nfo protein, including immediately opposed or closely spaced clusters. PUTR cleaved more efficiently than Nfo protein by a factor of approximately 1.7 or approximately 2 for DNA that had been gamma-irradiated in moderate or non-radioquenching conditions, respectively. This suggests that the DNA environment during irradiation affects the spectrum of cluster configurations. Further comparison of PUTR and Nfo protein cleavage may provide useful information on abasic cluster levels and configurations induced by ionizing radiation. PMID- 12087164 TI - Modulation of enzymatic activities of Escherichia coli DnaB helicase by single stranded DNA-binding proteins. AB - The modulation of enzymatic activities of Escherichia coli DnaB helicase by homologous and heterologous single-stranded DNA-binding proteins (SSBs) and its DNA substrates were analyzed. Although DnaB helicase can unwind a variety of DNA substrates possessing different fork-like structures, the rate of DNA unwinding was significantly diminished with substrates lacking a 3' fork. A 5 nt fork appeared to be adequate to attain the maximum rate of DNA unwinding. Efficient helicase action of DnaB requires the participation of SSBs. Studies involving heterologous SSBs demonstrated that they can stimulate the helicase activity of DnaB protein under certain conditions. However, this stimulation occurs in a manner distinctly different from that observed with cognate E.coli SSB. The E.coli SSB was found to stimulate the helicase activity over a wide range of SSB concentrations and was unique in its strong inhibition of single-stranded DNA dependent ATPase activity when uncoupled from the DNA helicase activity. In the presence of a helicase substrate, the ATPase activity of DnaB helicase remained uninhibited. Thus, E.coli SSB appears to coordinate and couple the ATPase activity to the DNA helicase activity by suppressing unproductive ATP hydrolysis by DnaB helicase. PMID- 12087165 TI - The mitochondrial DNA polymerase as a target of oxidative damage. AB - The mitochondrial respiratory chain is a source of reactive oxygen species (ROS) that are responsible for oxidative modification of biomolecules, including proteins. Due to its association with mitochondrial DNA, DNA polymerase gamma (pol gamma) is in an environment to be oxidized by hydrogen peroxide and hydroxyl radicals that may be generated in the presence of iron ions associated with DNA. We tested whether human pol gamma was a possible target of ROS with H2O2 and investigated the effect on the polymerase activities and DNA binding efficiency. A 1 h treatment with 250 microM H2O2 significantly inhibited DNA polymerase activity of the p140 subunit and lowered its DNA binding efficiency. Addition of p55 to the p140 catalytic subunit prior to H2O2 treatment offered protection from oxidative inactivation. Oxidatively modified amino acid residues in pol gamma resulting from H2O2 treatment were observed in vitro as well as in vivo, in SV40 transfected human fibroblasts. Pol gamma was detected as one of the major oxidized mitochondrial matrix proteins, with a detectable decline in polymerase activity. These results suggest pol gamma as a target of oxidative damage, which may result in a reduction in mitochondrial DNA replication and repair capacities. PMID- 12087166 TI - Footprinting, circular dichroism and UV melting studies on neomycin B binding to the packaging region of human immunodeficiency virus type-1 RNA. AB - We have studied the binding of neomycin to a 171mer RNA (psi-RNA) from the packaging region of the LAI strain of human immunodeficiency virus type-1, HIV-1 (LAI). The RNase I footprinting studies reveal that the primary binding site for the drug is in stem-loop 1, which contains the dimer initiation site of HIV-1. Loading this site with neomycin causes a structural change in the RNA, allowing nucleotides in the neighboring stem-loop 2 to participate in the drug site. Drug binding to secondary sites induces structural changes in other stem-loops of the RNA. Footprinting plots, showing cutting at a site as a function of drug concentration, were analyzed using a two-state model to obtain relative site specific binding constants. Circular dichroism measurements show that neomycin binding to psi-RNA changes the intensity of the strong negative CD band at 208 nm, confirming that neomycin induces structural changes. Melting studies of the RNA showed melting transitions in the absence of drug at 28.2, 37.2, 47.4, 55.5 and 60.8 degrees C. Only the first two were affected by drug binding, the reason for this being explained by our analysis. PMID- 12087167 TI - The identification of spermine binding sites in 16S rRNA allows interpretation of the spermine effect on ribosomal 30S subunit functions. AB - A photoreactive analogue of spermine, N1-azidobenzamidino (ABA)-spermine, was covalently attached after irradiation to Escherichia coli 30S ribosomal subunits or naked 16S rRNA. By means of RNase H digestion and primer extension, the cross linking sites of ABA-spermine in naked 16S rRNA were characterised and compared with those identified in 30S subunits. The 5' domain, the internal and terminal loops of helix H24, as well as the upper part of helix H44 in naked 16S rRNA, were found to be preferable binding sites for polyamines. Association of 16S rRNA with ribosomal proteins facilitated its interaction with photoprobe, except for 530 stem-loop nt, whose modification by ABA-spermine was abolished. Association of 30S with 50S subunits, poly(U) and AcPhe-tRNA (complex C) further altered the susceptibility of ABA-spermine cross-linking to 16S rRNA. Complex C, modified in its 30S subunit by ABA-spermine, reacted with puromycin similarly to non photolabelled complex. On the contrary, poly(U)-programmed 70S ribosomes reconstituted from photolabelled 30S subunits and untreated 50S subunits bound AcPhe-tRNA more efficiently than untreated ribosomes, but were less able to recognise and reject near cognate aminoacyl-tRNA. The above can be interpreted in terms of conformational changes in 16S rRNA, induced by the incorporation of ABA spermine. PMID- 12087168 TI - Conformational change of Escherichia coli initiator methionyl-tRNA(fMet) upon binding to methionyl-tRNA formyl transferase. AB - The specific formylation of initiator methionyl-tRNA (Met-tRNA) by methionyl-tRNA formyltransferase (MTF) is important for the initiation of protein synthesis in Escherichia coli. The determinants for formylation are located in the acceptor stem and in the dihydrouridine (D) stem of the initiator tRNA (tRNA(fMet)). Here, we have used ethylation interference analysis to study the interactions between the Met-tRNA(fMet) and MTF in solution. We have identified three clusters of phosphates in the tRNA that, when ethylated, interfere with binding of MTF. Interference due to ethylation of phosphates in the acceptor stem and in the D stem is most likely due to the close proximity of the protein as seen in the crystal structure of the MTF.fMet-tRNA(fMet) complex. The third cluster of phosphates, whose ethylation interferes with binding of MTF, is dispersed along the anticodon stem, which is distal to the sites of tRNA protein contacts. Interestingly, these latter positions correspond to sites of increased cleavages by RNase V1 in RNA footprinting experiments. Together, these results suggest that in addition to the protein, which binds to the substrate tRNA in an induced fit mechanism, the tRNA also undergoes induced structural changes during its binding to MTF. PMID- 12087170 TI - Effects of double-strand break repair proteins on vertebrate telomere structure. AB - Although telomeres are not recognized as double-strand breaks (DSBs), some DSB repair proteins are present at telomeres and are required for telomere maintenance. To learn more about the telomeric function of proteins from the homologous recombination (HR) and non-homologous end joining pathways (NHEJ), we have screened a panel of chicken DT40 knockout cell lines for changes in telomere structure. In contrast to what has been observed in Ku-deficient mice, we found that Ku70 disruption did not result in telomere-telomere fusions and had no effect on telomere length or the structure of the telomeric G-strand overhang. G overhang length was increased by Rad51 disruption but unchanged by disruption of DNA-PKcs, Mre11, Rad52, Rad54, XRCC2 or XRCC3. The effect of Rad51 depletion was unexpected because gross alterations in telomere structure have not been detected in yeast HR mutants. Thus, our results indicate that Rad51 has a previously undiscovered function at vertebrate telomeres. They also indicate that Mre11 is not required to generate G-overhangs. Although Mre11 has been implicated in overhang generation, overhang structure had not previously been examined in Mre11 deficient cells. Overall our findings indicate that there are significant species specific differences in the telomeric function of DSB repair proteins. PMID- 12087169 TI - Internal ribosome entry site-mediated translation of Smad5 in vivo: requirement for a nuclear event. AB - Smad5 is thought to relay signals of the bone morphogenetic protein pathway. The 5' untranslated region (5'UTR) of human Smad5 mRNA is long, has the potential to form secondary structures and contains five AUG codons. Here we show that the 5'UTR of Smad5 contains an internal ribosome entry site (IRES) located within 100 nt of the 3' end of the 5'UTR. The Smad5 IRES was 4-8-fold more active than the poliovirus IRES in C2C12 cells, which have osteoblastic differentiation ability, but was 5-10-fold less active than the poliovirus IRES in 293T cells. When an in vitro transcript of a dicistronic Smad5 IRES construct was transfected into C2C12 cells, the Smad5 IRES was not able to stimulate the translation of the downstream cistron, although the cap-dependent translation of the upstream cistron was efficient. In contrast, the poliovirus IRES in a dicistronic in vitro transcript was able to stimulate the translation of the downstream cistron to a similar extent as in the case of transfection of the corresponding dicistronic DNA construct. These results suggest that Smad5 IRES activity displays cell specificity and that some as yet unidentified nuclear event may be required for efficient Smad5 IRES-driven translation initiation. PMID- 12087171 TI - Minisatellite instability at the Adh locus reveals somatic polymorphism in amphioxus. AB - Amphioxus (subphylum Cephalochordata) is the closest living relative to vertebrates and widely used for phylogenetic analyses of vertebrate gene evolution. Amphioxus genes are highly polymorphic, but the origin and nature of this variability is unknown. We have analyzed the alcohol dehydrogenase locus (Adh3) in two amphioxus species (Branchiostoma lanceolatum and Branchiostoma floridae) and found that genetic variation is related to repetitive DNA sequences, mainly minisatellites. Small pool-PCR assays indicated that allelic variants are generated by minisatellite instability. We conclude that the generation of new forms was not preferentially linked to germline processes but rather to somatic events leading to mosaic adult animals. Furthermore, most Adh minisatellites belong to a novel class, which we have named mirages. Their distinctive feature is that the repeat subunit spans the exon-intron boundaries and generates potential duplications of the splice sites. However, splicing may not be compromised as no aberrant mRNA variants were detected. PMID- 12087172 TI - Synthesis and application of charge-modified dye-labeled dideoxynucleoside-5' triphosphates to 'direct-load' DNA sequencing. AB - A novel series of charge-modified, dye-labeled 2',3'-dideoxynucleoside triphosphate terminators were synthesized and evaluated as reagents for DNA sequencing. These terminators possess an advantage over existing reagents in that no purification is required to remove unreacted nucleotide or associated breakdown products prior to electrophoretic separation of the sequencing fragments. This obviates the need for a time consuming post-reaction work up, allowing direct loading of DNA sequencing reaction mixtures onto a slab gel. Thermo Sequenase II DNA polymerase poorly incorporates the charge-modified terminators compared with regular dye-labeled terminators. However, extending the linker arm between dye and nucleotide and using a mutant form of a related DNA polymerase can in part mitigate the decrease in substrate efficiency. We also present evidence that these charge-modified terminators can relieve gel compression artefacts when used with dGTP in sequencing reactions. PMID- 12087173 TI - Co-expression pattern from DNA microarray experiments as a tool for operon prediction. AB - The prediction of operons, the smallest unit of transcription in prokaryotes, is the first step towards reconstruction of a regulatory network at the whole genome level. Sequence information, in particular the distance between open reading frames, has been used to predict if adjacent Escherichia coli genes are in an operon. While appreciably successful, these predictions need to be validated and refined experimentally. As a growing number of gene expression array experiments on E.coli became available, we investigated to what extent they could be used to improve and validate these predictions. To this end, we examined a large collection of published microarry data. The correlation between expression ratios of adjacent genes was used in a Bayesian classification scheme to predict whether the genes are in an operon or not. We found that for the genes whose expression levels change significantly across the experiments in the data set, the currently available gene expression data allowed a significant refinement of the sequenced based predictions. We report these co-expression correlations in an E.coli genomic map. For a significant portion of gene pairs, however, the set of array experiments considered did not contain sufficient information to determine whether they are in the same transcriptional unit. This is not due to unreliability of the array data per se, but to the design of the experiments analyzed. In general, experiments that perturb a large number of genes offer more information for operon prediction than confined perturbations. These results provide a rationale for conducting expression studies comparing conditions that cause global changes in gene expression. PMID- 12087174 TI - DNA bending and unwinding due to the major 1,2-GG intrastrand cross-link formed by antitumor cis-diamminedichloroplatinum(II) are flanking-base independent. AB - Antitumor cisplatin [cis-diamminedichloroplatinum(II)] forms on DNA predominantly intrastrand cross-links between neighboring purine residues. Several discoveries suggested that the toxicity of cisplatin originated from these lesions. The formation of 1,2-GG intrastrand cross-link of cisplatin leads to marked conformational alterations in DNA including a directional, rigid bend toward the major groove and local unwinding. These altered structures attract various cellular proteins. This phenomenon has been postulated to mediate antitumor properties of cisplatin. Importantly, the binding affinity of several proteins that specifically recognize 1,2-GG intrastrand cross-link to platinated DNA is modulated by the nature of the base pairs that immediately flank the platinated d(GpG) site. However, the influence of sequence context on DNA bending and unwinding due to the formation of the 1,2-GG intrastrand cross-link has not been extensively investigated. In the present study we have employed electrophoretic retardation (phasing) assay to analyze bending and unwinding induced by the single, site-specific 1,2-GG intrastrand cross-link immediately flanked by various bases formed by cisplatin in nine oligodeoxyribonucleotide duplexes. The results indicate that bending and unwinding of DNA as a consequence of the formation of the major adduct of cisplatin is, in the first approximation, independent of the base pairs flanking the platinated d(GpG) site. PMID- 12087175 TI - The chAB4 and NF1-related long-range multisequence DNA families are contiguous in the centromeric heterochromatin of several human chromosomes. AB - We have investigated the large-scale organization of the human chAB4-related long range multisequence family, a low copy-number repetitive DNA located in the pericentromeric heterochromatin of several human chromosomes. Analysis of genomic clones revealed large-scale ( approximately 100 kb or more) sequence conservation in the region flanking the prototype chAB4 element. We demonstrated that this low copy-number family is connected to another long-range repeat, the NF1-related (PsiNF1) multisequence. The two DNA types are joined by an approximately 2 kb long tandem repeat of a 48-bp satellite. Although the chAB4- and NF1-like sequences were known to have essentially the same chromosomal localization, their close association is reported here for the first time. It indicates that they are not two independent long-range DNA families, but are parts of a single element spanning approximately 200 kb or more. This view is consistent both with their similar chromosomal localizations and the high levels of sequence conservation among copies found on different chromosomes. We suggest that the master copy of the linked chAB4-PsiNF1 DNA segment appeared first on the ancestor of human chromosome 17. PMID- 12087176 TI - Overlapping of MINK and CHRNE gene loci in the course of mammalian evolution. AB - Overlapping of genes, especially in an anti-parallel fashion, is quite rare in eukaryotic genomes. We have found a rare instance of exon overlapping involving CHRNE and MINK gene loci on chromosome 17 in humans. CHRNE codes for the epsilon subunit of the nicotinic acetylcholine receptor (AChRepsilon) whereas MINK encodes a serine/threonine kinase belonging to the GCK family. To elucidate the evolutionary trail of this gene overlapping event, we examined the genomes of a number of primates and found that mutations in the polyadenylation signal of the CHRNE gene in early hominoids led to the overlap. Upon extending this analysis to genomes of other orders of placental mammals, we observed that the overlapping occurred at least three times independently during the course of mammalian evolution. Because CHRNE and MINK are differentially expressed, the potentially hazardous mutations responsible for the exon overlap seem to have escaped evolutionary pressures by differential temporo-spatial expression of the two genes. PMID- 12087178 TI - Analysis of cell-cycle gene expression in Saccharomyces cerevisiae using microarrays and multiple synchronization methods. AB - Microarray analysis of gene expression during the yeast division cycle has led to the proposal that a significant number of genes in Saccharomyces cerevisiae are expressed in a cell-cycle-specific manner. Four different methods of synchronization were used for cell-cycle analysis. Randomized data exhibit periodic patterns of lesser strength than the experimental data. Thus the cyclicities in the expression measurements in the four experiments presented do not arise from chance fluctuations or noise in the data. However, when the degree of cyclicity for genes in different experiments are compared, a large degree of non-reproducibility is found. Re-examining the phase timing of peak expression, we find that three of the experiments (those using alpha-factor, CDC28 and CDC15 synchronization) show consistent patterns of phasing, but the elutriation synchrony results demonstrate a different pattern from the other arrest-release synchronization methods. Specific genes can show a wide range of cyclical behavior between different experiments; a gene with high cyclicity in one experiment can show essentially no cyclicity in another experiment. The elutriation experiment, possibly being the least perturbing of the four synchronization methods, may give the most accurate characterization of the state of gene expression during the normal, unperturbed cell cycle. Under this alternative explanation, the observed cyclicities in the other three experiments are a stress response to synchronization, and may not reproduce in unperturbed cells. PMID- 12087177 TI - The p120(ctn)-binding partner Kaiso is a bi-modal DNA-binding protein that recognizes both a sequence-specific consensus and methylated CpG dinucleotides. AB - The p120(ctn)-binding partner Kaiso is a new member of the POZ-zinc finger family of transcription factors implicated in development and cancer. To understand the role of Kaiso in gene regulation and p120(ctn)-mediated signaling and adhesion, we sought to identify Kaiso-specific DNA binding sequences and potential target genes. Here we demonstrate that Kaiso is a dual specificity DNA-binding protein that recognizes the specific consensus sequence TCCTGCNA as well as methyl-CpG dinucleotides. A minimal core sequence CTGCNA was identified as sufficient for Kaiso binding. Two copies of the Kaiso-binding site are present in the human and murine matrilysin promoters, implicating matrilysin as a candidate target gene for Kaiso. In electrophoretic mobility shift assays, matrilysin promoter-derived oligonucleotide probes formed a complex with GST-Kaiso fusion proteins possessing the zinc finger domain but not with fusion proteins lacking the zinc fingers. We further determined that only Kaiso zinc fingers 2 and 3 were necessary and sufficient for sequence-specific DNA binding. Interestingly, Kaiso also possesses a methyl-CpG-dependent DNA-binding activity distinct from its sequence-specific DNA binding. However, Kaiso has a higher affinity for the TCCTGCNA consensus than for the methyl-CpG sites. Furthermore, the DNA-binding ability of Kaiso with either recognition site was inhibited by p120(ctn). Kaiso thus appears to have two modes of DNA binding and transcriptional repression, both of which may be modulated by its interaction with the adhesion cofactor p120(ctn). PMID- 12087179 TI - On the epigenetic regulation of the human reelin promoter. AB - Reln mRNA and protein levels are reduced by approximately 50% in various cortical structures of post-mortem brain from patients diagnosed with schizophrenia or bipolar illness with psychosis. To study mechanisms responsible for this down regulation, we have analyzed the promoter of the human reelin gene. We show that the reelin promoter directs expression of a reporter construct in multiple human cell types: neuroblastoma cells (SHSY5Y), neuronal precursor cells (NT2), differentiated neurons (hNT) and hepatoma cells (HepG2). Deletion constructs confirmed the presence of multiple elements regulating Reln expression, although the promoter activity is promiscuous, i.e. activity did not correlate with expression of the endogenous gene as reflected in terms of reelin mRNA levels. Co transfection of the -514 bp human reelin promoter with either Sp1 or Tbr1 demonstrated that these transcription factors activate reporter expression by 6- and 8.5-fold, respectively. Within 400 bp of the RNA start site there are 100 potential CpG targets for DNA methylation. Retinoic acid (RA)-induced differentiation of NT2 cells to hNT neurons was accompanied by increased reelin expression and by the appearance of three DNase I hypersensitive sites 5' to the RNA start site. RA-induced differentiation was also associated with demethylation of the reelin promoter. To test if methylation silenced reelin expression, we methylated the promoter in vitro prior to transfection. In addition, we treated NT2 cells with the methylation inhibitor aza-2'-deoxycytidine and observed a 60 fold increase in reelin mRNA levels. The histone deacetylase inhibitors trichostatin A (TSA) and valproic acid also induced expression of the endogenous reelin promoter, although TSA was considerably more potent. These findings indicate that one determinant responsible for regulating reelin expression is the methylation status of the promoter. Our data also raise the interesting possibility that the down-regulation of reelin expression documented in psychiatric patients might be the consequence of inappropriate promoter hypermethylation. PMID- 12087180 TI - Premature termination of RNA polymerase II mediated transcription of a seed protein gene in Schizosaccharomyces pombe. AB - The poly(A) signal and downstream elements with transcriptional pausing activity play an important role in termination of RNA polymerase II transcription. We show that an intronic sequence derived from the plant seed protein gene (AmA1) specifically acts as a transcriptional terminator in the fission yeast, Schizosaccharomyces pombe. The 3'-end points of mRNA encoded by the AmA1 gene were mapped at different positions in S.pombe and in native cells of Amaranthus hypochondriacus. Deletion analyses of the AmA1 intronic sequence revealed that multiple elements essential for proper transcriptional termination in S.pombe include two site-determining elements (SDEs) and three downstream sequence elements. RT-PCR analyses detected transcripts up to the second SDE. This is the first report showing that the highly conserved mammalian poly(A) signal, AAUAAA, is also functional in S.pombe. The poly(A) site was determined as Y(A) both in native and heterologous systems but at different positions. Deletion of these cis elements abolished 3'-end processing in S.pombe and a single point mutation in this motif reduced the activity by 70% while enhancing activity at downstream SDE. These results indicate that the bipartite sequence elements as signals for 3'-end processing in fission yeast act in tandem with other cis-acting elements. A comparison of these elements in the AmA1 intron that function as a transcriptional terminator in fission yeast with that of its native genes showed that both require an AT-rich distal and proximal upstream element. However, these sequences are not identical. Transcription run-on analysis indicates that elongating RNA polymerase II molecules accumulate over these pause signals, maximal at 611-949 nt. Furthermore, we demonstrate that the AmA1 intronic terminator sequence acts in a position-independent manner when placed within another gene. PMID- 12087181 TI - Parameter optimized surfaces (POPS): analysis of key interactions and conformational changes in the ribosome. AB - We present a new method for the calculation of solvent accessible surface areas at the atomic and residue levels, which we call parameter optimized surfaces (POPS-A and POPS-R ). Atomic and residue areas (the latter simulated with a single sphere centered at the C(alpha)s atom for amino acids and at the P atom for nucleotides) have been optimized versus accurate all-atoms methods. We concentrated on an analytical formula for the approximation of solvent accessibilities. The formula is simple, easily derivable and fast to compute, therefore it is practical for use in molecular dynamics simulations as an approximation to the first solvation shell. The residue based approach POPS-R has been derived as a useful tool for the analysis of large macromolecular assemblies like the ribosome, and is especially suited for use in refinement of low resolution structures. The structures of the 70S, 50S and 30S ribosomes have been analyzed in detail and most of the interactions within the subunits and at their interfaces were clearly identified. Some interesting differences between 30S alone and within the 70S have been highlighted. Owing to the presence of the P tRNA in the 70S ribosome, localized conformational rearrangements occur within the subunits, exposing Arg and Lys residues to negatively charged binding sites of P-tRNA. POPS-R also allows for estimates of the loss of free energy of solvation upon complex formation, particularly useful in designing new protein RNA complexes and in suggesting more focused experimental work. PMID- 12087182 TI - Pentamidine inhibits catalytic activity of group I intron Ca.LSU by altering RNA folding. AB - The antimicrobial agent pentamidine inhibits the self-splicing of the group I intron Ca.LSU from the transcripts of the 26S rRNA gene of Candida albicans, but the mechanism of pentamidine inhibition is not clear. We show that preincubation of the ribozyme with pentamidine enhances the inhibitory effect of the drug and alters the folding of the ribozyme in a pattern varying with drug concentration. Pentamidine at 25 microM prevents formation of the catalytically active F band conformation of the precursor RNA and alters the ribonuclease T1 cleavage pattern of Ca.LSU RNA. The effects on cleavage suggest that pentamidine mainly binds to specific sites in or near asymmetric loops of helices P2 and P2.1 on the ribozyme, as well as to the tetraloop of P9.2 and the loosely paired helix P9, resulting in an altered structure of helix P7, which contains the active site. Positively charged molecules antagonize pentamidine inhibition of catalysis and relieve the drug effect on ribozyme folding, suggesting that pentamidine binds to a magnesium binding site(s) of the ribozyme to exert its inhibitory effect. PMID- 12087184 TI - Structural perturbations in DNA caused by bis-intercalation of ditercalinium visualised by atomic force microscopy. AB - Atomic force microscopy (AFM) has been used to examine perturbations in the tertiary structure of DNA induced by the binding of ditercalinium, a DNA bis intercalator with strong anti-tumour properties. We report AFM images of plasmid DNA of both circular and linearised forms showing a difference in the formation of supercoils and plectonemic coils caused at least in part by alterations in the superhelical stress upon bis-intercalation. A further investigation of the effects of drug binding performed with 292 bp mixed-sequence DNA fragments, and using increment in contour length as a reliable measure of intercalation, revealed saturation occurring at a point where sufficient drug was present to interact with every other available binding site. Moment analysis based on the distribution of angles between segments along single DNA molecules showed that at this level of bis-intercalation, the apparent persistence length of the molecules was 91.7 +/- 5.7 nm, approximately twice as long as that of naked DNA. We conclude that images of single molecules generated using AFM provide a valuable supplement to solution-based techniques for evaluation of physical properties of biological macromolecules. PMID- 12087183 TI - Functional analysis of the transcription factor ER71 and its activation of the matrix metalloproteinase-1 promoter. AB - The ETS transcription factor family is characterized by a conserved ETS DNA binding domain and its members have been implicated in a plethora of biological processes, including development, cell transformation and metastasis. ER71 is a testis-specific ETS protein that is not homologous to any other protein outside its ETS domain, suggesting that it fulfills a unique physiological role. Here, we report that ER71 is a constitutively nuclear protein whose intracellular localization is dependent on a portion of the ETS domain, namely ER71 amino acids 276-315. Furthermore, the DNA binding activity is intramolecularly regulated, as the N-terminus of ER71 has a negative effect on DNA binding while the C-terminus dramatically enhances this activity. We also demonstrate that ER71 possesses an extremely potent N-terminal transactivation domain comprised of amino acids 1 157. Finally, we show that ER71 is capable of directly activating both an E74 site-driven and the matrix metalloproteinase-1 promoter. Altogether, these data represent the first functional characterization of ER71, which may perform important functions in the developing and adult testis as well as in testicular germ cell tumorigenesis. PMID- 12087185 TI - Origin and fate of repeats in bacteria. AB - We investigated 53 complete bacterial chromosomes for intrachromosomal repeats. In previous studies on eukaryote chromosomes, we proposed a model for the dynamics of repeats based on the continuous genesis of tandem repeats, followed by an active process of high deletion rate, counteracted by rearrangement events that may prevent the repeats from being deleted. The present study of long repeats in the genomes of Bacteria and Archaea suggests that our model of interspersed repeats dynamics may apply to them. Thus the duplication process might be a consequence of very ancient mechanisms shared by all three domains. Moreover, we show that there is a strong negative correlation between nucleotide composition bias and the repeat density of genomes. We hypothesise that in highly biased genomes, non-duplicated small repeats arise more frequently by random effects and are used as primers for duplication mechanisms, leading to a higher density of large repeats. PMID- 12087187 TI - Studies on the synthesis of a G-rich octaoligoisonucleotide (isoT)2(isoG)4(isoT)2 by the phosphotriester approach and its formation of G-quartet structure. AB - The octaoligoisonucleotide (isoT)2(isoG)4(isoT)2 (I), consisting of isonucleoside units 6'-O-allyl-4'-deoxy-4'-(nucleobase)-2',5'-anhydro-L-mannitol, was synthesized by the phosphotriester approach in solution phase. Based on CD spectra and capillary electrophoresis, it was confirmed that iso-oligomer I could form a parallel intermolecular G-quadruplex structure. K+, Na+ and Li+ can prompt the formation of G-quartet structures and stabilize them. The effective order of these cations is K+ > Na+ > Li+. PMID- 12087188 TI - A pH-jump approach for investigating secondary structure refolding kinetics in RNA. AB - It has been shown that premature translation of the plasmid-mediated toxin in hok/sok of plasmid R1 and pnd/pndB of plasmid R483 is prevented during transcription of the hok and pnd mRNAs by the formation of metastable hairpins at the 5'-end of the mRNA. Here, an experimental approach is presented, which allows the accurate measurement of the refolding kinetics of the 5'-end RNA fragments in vitro without chemically modifying the RNA. The method is based on acid denaturation followed by a pH-jump to neutral pH as a novel way to trap kinetically favoured RNA secondary structures, allowing the measurement of a wide range of biologically relevant refolding rates, with or without the use of standard stopped-flow equipment. The refolding rates from the metastable to the stable conformation in both the hok74 and pnd58 5'-end RNA fragments were determined by using UV absorbance changes corresponding to the structural rearrangements. The measured energy barriers showed that the refolding path does not need complete unfolding of the metastable structures before the formation of the final structures. Two alternative models of such a pathway are discussed. PMID- 12087189 TI - Characterization of the repetitive sequences in a 200-kb region around the rice waxy locus: diversity of transposable elements and presence of veiled repetitive sequences. AB - Repetitive genomic sequences might have various structural features and properties distinct from those of the known transposable elements (TE). Here, the content and properties of the repetitive sequences present in a 200-kb region around the rice waxy locus were analyzed using the available rice genomic database. In our previous Southern blotting analysis, 70% of the segments in this region showed smeared patterns, but according to the present database analysis, the proportion of repetitive sequences in this region was only 15%. The repetitive segments in this 200-kb region comprised 75 repetitive sequences that we classified into 46 subfamilies: 21 subfamilies were known TEs or repetitive sequences and 25 subfamilies consisted of newly identified TEs or novel types of repetitive sequences. The region contains no long terminal repeat (LTR) retrotransposable elements, but miniature inverted repeat transposable elements (MITEs) constituted a major class among the elements identified. These MITEs showed remarkable structural divergence: 12 elements were found to be new members of known MITE superfamilies, while five elements had novel terminal structures, and did not belong to any known TE families. Interestingly, about 10% of the repetitive sequences, including virus-like sequences did not have any of the usual characteristics of TEs, suggesting that a certain proportion of repetitive sequences that might not share the transpositional mechanisms of known elements are dispersed in the compact rice genome. PMID- 12087186 TI - The role of DNA polymerase beta in determining sensitivity to ionizing radiation in human tumor cells. AB - Lethal lesions after ionizing radiation are thought to be mainly unrepaired or misrepaired DNA double-strand breaks, ultimately leading to lethal chromosome aberrations. However, studies with radioprotectors and repair inhibitors indicate that single-strand breaks, damaged nucleotides or abasic sites can also influence cell survival. This paper reports on studies to further define the role of base damage and base excision repair on the radiosensitivity of human cells. We retrovirally transduced human tumor cells with a dominant negative form of DNA polymerase beta, comprising the 14 kDa DNA-binding domain of DNA polymerase beta but lacking polymerase function. Radiosensitization of two human carcinoma cell lines, A549 and SQD9, was observed, achieving dose enhancement factors of 1.5 1.7. Sensitization was dependent on expression level of the dominant negative and was seen in both single cell clones and in unselected virally transduced populations. Sensitization was not due to changes in cell cycle distribution. Little or no sensitization was seen in G(1)-enriched populations, indicating cell cycle specificity for the observed sensitization. These results contrast with the lack of effect seen in DNA polymerase beta knockout cells, suggesting that polDN also inhibits the long patch, DNA polymerase beta-independent repair pathway. These data demonstrate an important role for BER in determining sensitivity to ionizing radiation and might help identify targets for radiosensitizing tumor cells. PMID- 12087190 TI - Characterization of two non-homoeologous nuclear genes encoding mitochondrial alternative oxidase in common wheat. AB - Mitochondrial alternative oxidase (AOX) is the terminal oxidase responsible for cyanide-insensitive and salicylhydroxamic acid-sensitive respiration. We have isolated two non-homoeologous genes (Waox1a and Waox1c) encoding AOX proteins from common wheat (Triticum aestivum L.). These two genes were orthologous to rice AOX1a and AOX1c, and their exon/intron structure was conserved, as it is in most other plant AOX genes. Southern blot analysis indicated that both Waox1a and Waox1c were located in at least three homoeologous loci and that additional AOX genes with lower homology were present in the genome of common wheat. The Waox1a and Waox1c loci were respectively assigned to the homoeologous group 2 and 6 chromosomes. The steady-state level of Waox1a and Waox1c transcripts increased under cold stress, while only that of Waox1a was increased by cyanide treatment. PMID- 12087192 TI - Search for species-specific mating signal in courtship songs of sympatric sibling species, Drosophila ananassae and D. pallidosa. AB - Sexual isolation is one of the most important mechanisms that may lead to speciation. Drosophila ananassae and D. pallidosa are useful for the study of sexual isolation, because of their sympatric distribution and no postmating isolation between them. Courtship songs are considered to play a crucial role in sexual isolation between D. ananassae and D. pallidosa. We recorded and analyzed male courtship songs of D. ananassae and D. pallidosa for eight and four geographical strains, respectively. Courtship behaviors of the two species were consistent with those previously described, however, male's middle leg shaking, which had not before been described, was observed in both species. Males sing by wing vibration only during courtship. Their song oscillograms were distinct between species, but those of conspecific strains were very similar, in spite of their different geographical derivation. We found species-specificity in burst length, pulse length, cycle number in a pulse, and frequency spectra of bursts; these results suggest that these song parameters may play a role in mate recognition that enforces their sexual isolation. The specific values of interpulse interval, cycle number in a pulse and intrapulse frequency were involved with the determination of specificity in frequency spectra of bursts. We discussed the possibility that the specific frequency spectra of bursts are recognized by females as the species-specific signal rather than each parameter individually. PMID- 12087191 TI - A single-nucleotide mutation in a gene encoding S-adenosylmethionine synthetase is associated with methionine over-accumulation phenotype in Arabidopsis thaliana. AB - Met-overaccumulating mutants provide a powerful genetic tool for examining both the regulation of the Met biosynthetic pathway and in vivo developmental responses of gene expression to altered Met levels. We have previously reported the identification of two Arabidopsis thaliana Met over-accumulation (mto) mutants, mto1-1 and mto2-1, that carry mutations in the genes encoding cystathionine gamma-synthase (CGS) and threonine synthase (TS), respectively. A third mutant, mto3-1, has recently been reported to carry a mutation in the gene encoding S-adenosylmethionine synthetase 3 (SAMS3). Here, we report the isolation of a new ethionine-resistant A. thaliana mutant that over-accumulates soluble Met approximately 20-fold in young rosettes. The causal mutation was determined to be a single, recessive mutation that was mapped to chromosome 3. Sequence analysis identified a single nucleotide change in the gene encoding SAMS3 that was distinct from the mto3-1 mutation and altered the amino acid sequence of the enzyme active site. This mutation was therefore referred to as mto3-2. Although Met over-accumulation in the mto3-2 mutant was similar to that in the mto2-1 mutant, CGS mRNA levels did not respond to the mto3-2 mutation and were similar to that in equivalent wild-type plants. PMID- 12087193 TI - Molecular phylogeny of Japanese Leporidae, the Amami rabbit Pentalagus furnessi, the Japanese hare Lepus brachyurus, and the mountain hare Lepus timidus, inferred from mitochondrial DNA sequences. AB - We determined mitochondrial 12S ribosomal RNA (rRNA) and cytochrome b (cyt b) gene sequences in three leporid species of Japan, the Amami rabbit Pentalagus furnessi from the Ryukyu Islands, the Japanese hare Lepus brachyurus from Honshu, and a Japanese form of the mountain hare Lepus timidus ainu from Hokkaido. We compared the sequences with those of other taxa of leporids available in databases. Phylogenetic trees of the 12S rRNA gene sequences indicated that the lineage of P. furnessi diversified during the generic radiation of the leporids at an ancient time, which was estimated to have been the middle Miocene. Cyt-b gene trees revealed that the lineage of L. brachyurus branched off at an early stage in the speciation of Lepus, probably at the beginning of the Pliocene. The cyt b sequences of L. t. ainu were somewhat distinct from those of continental conspecific populations; this lineage divergence is likely to have occurred during the middle or late Pleistocene. The results show that the three regions of the Japanese archipelago, Ryukyu, Honshu-Shikoku-Kyushu, and Hokkaido, now preserve their own leporid taxa, each with a different extent of genetic endemicity. It is possible that the zoogeographic traits of the Japanese leporids are a consequence of the evolutionary dynamics of leporids in East Asia, in that the radiation centers of leporids are likely to have shifted from tropical, through temperate, to arctic zones. PMID- 12087194 TI - A new mutation of the ATP-binding cassette, sub-family C, member 2 (ABCC2) gene in a Japanese patient with Dubin-Johnson syndrome. AB - Dubin-Johnson syndrome (DJS) is an inherited disorder characterized by conjugated hyperbilirubinemia and is caused by mutations of the canalicular multispecific organic anion transporter (cMOAT)/ multidrug resistance protein 2 (MRP2)/ ATP binding cassette, sub-family C, member 2 (ABCC2) gene. The ABCC2 protein is located in the apical membrane of hepatocytes, and known mutations of this gene cause impaired maturation and trafficking of the mutated protein from the endoplasmic reticulum (ER) to the Golgi complex. We have characterized the ABCC2 gene in a Japanese DJS patient by polymerase chain reaction and DNA sequencing, resulting in the identification of two mutations. One mutation, 1815+2 (T>A) in the splice donor site of intron 13, has already been reported. However, we have identified a novel nonsense mutation consisting of a (C>T) transition at nucleotide 3928 in exon 28. PMID- 12087195 TI - A recursive procedure to compute the gametic relationship matrix and its inverse for marked QTL clusters. AB - Recently, a variety of mixed linear models have been proposed for marker-assisted prediction of the effects of quantitative trait loci (QTLs) in outbred populations of animals. One of them addresses the effects of a cluster of linked QTLs, or those of a particular chromosomal segment, marked by DNA marker(s) and requires that the inverse of the corresponding gametic relationship matrix whose elements are the conditional expected values of the identity-by-descent (IBD) proportions between gametes for individuals be evaluated. Here, for a model of this type, utilizing the property of the IBD set and using the information on the joint gametogenesis processes at the flanking marker loci, we present a recursive method to systematically calculate the elements of the gametic relationship matrix and its inverse. A numerical example is given to illustrate the proposed computing procedure. PMID- 12087196 TI - Ty1-copia group retrotransposons in persimmon (Diospyros kaki Thunb.). AB - We cloned and characterized Ty1-copia group retrotransposons in persimmon (Diospyros kaki Thunb.). Genomic DNA or methyl jasmonate (MJA)-treated cDNA were used as templates to amplify the reverse transcriptase region of Ty1-copia group retrotransposons. About 280 bp fragments were amplified and cloned, and 97 clones were sequenced. Forty-nine clones included frameshift or the stop codon, or both. Among 48 clones containing complete reading frames, 42 clones had unique nucleotide sequences. Alignment and phylogenetic analysis of putative amino acid sequences in the 42 clones indicated that these clones (named Tdk; retroTransposon in Diospyros kaki) fell into seven subgroups and six ungrouped sequences, indicating high sequence heterogeneity in Tdk clones. Phylogenetic analysis comparing unrelated plant species shows that some Tdk clones are more closely related to Ty1-copia group retrotransposons in the orders Solanales and Sapindales rather than to other Tdk clones. Southern blot analysis using Tdk2B, Tdk4c, Tdk6Ac, Tdk12K and Tdk13G clones as probes showed that persimmon and its related species, D. lotus, D. lotus var. glabba, D. oleifera, D. rhombifolia and D. virginiana, contained multiple Tdk-like sequences, indicating that homologous elements exist in other Diospyros species. PMID- 12087197 TI - The efficacy of metallic stent placement in the treatment of colorectal obstruction. AB - OBJECTIVE: To evaluate the efficacy of newly designed covered and non-covered coated colorectal stents for colonic decompression. MATERIALS AND METHODS: Twenty six patients, (15 palliative cases and 11 preoperative) underwent treatment for the relief of colorectal obstruction using metallic stents positioned under fluoroscopic guidance. In 24 of the 26, primary colorectal carcinoma was diagnosed, and in the remaining two, recurrent colorectal carcinoma. Twenty-one patients were randomly selected to receive either a type A or type B stent; for the remaining five, type C was used. Type A, an uncovered nitinol wire stent, was lightly coated to ensure structural integrity. Type B (flare type) and C (shoulder type) stents were polyurethane covered and their diameter was 24 and 26 mm, respectively. The rates of technical success, clinical success, and complications were analyzed using the chi-square test, and to analyse the mean period of patency, the Kaplan-Meier method was used. RESULTS: Thirty of 31 attempted placements in 26 patients were successful, with a technical success rate of 96.8% (30/31) and a clinical success rate of 80.0% (24/30). After clinically successful stent placement, bowel decompression occurred within 1-4 (mean, 1.58+/-0.9) days. Five of six clinical failures involved stent migration and one stent did not expand after successful placement. In the preoperative group, 11 stents, one of which migrated, were placed in ten patients, in all of whom bowel preparation was successful. In the palliative group, 19 stents were placed in 15 patients. The mean period of patency was 96.25+/-105.12 days: 146.25+/-112.93 for type-A, 78.82+/-112.26 for type-B, and 94.25+/-84.21 for type C. Complications associated with this procedure were migration (n=6, 20%), pain (n=4, 13.3%), minor bleeding (n=5, 16.7%), incomplete expansion (n=1, 3.3%), and tumor ingrowth (n=1, 3.3%). The migration rate was significantly higher in the type-B group than in other groups (p=0.038). CONCLUSION: Newly designed covered and non-covered metallic stents of a larger diameter are effective for the treatment of colorectal obstruction. The migration rate of covered stents with flaring is higher than that of other types. For evaluation of the ideal stent configuration for the relief of colorectal obstruction, a clinical study involving a larger patient group is warranted. PMID- 12087198 TI - Optimal pulse sequence for ferumoxides-enhanced MR imaging used in the detection of hepatocellular carcinoma: a comparative study using seven pulse sequences. AB - OBJECTIVE: To identify the optimal pulse sequence for ferumoxides-enhanced magnetic resonance (MR) imaging in the detection of hepatocelluar carcinomas (HCCs). MATERIALS AND METHODS: Sixteen patients with 25 HCCs underwent MR imaging following intravenous infusion of ferumoxides. All MR studies were performed on a 1.5-T MR system, using a phased-array coil. Ferumoxides (Feridex IV) at a dose of 15 micromol/Kg was slowly infused intravenously, and axial images of seven sequences were obtained 30 minutes after the end of infusion. The MR protocol included fast spin-echo (FSE) with two echo times (TR3333-8571/TE18 and 90-117), singleshot FSE (SSFSE) with two echo times (TRinfinity/TE39 and 98), T2*-weighted gradient- recalled acquisition in the steady state (GRASS) (TR216/TE20), T2* weighted fast multiplanar GRASS (FMPGR) (TR130/TE8.4-9.5), and T2*-weighted fast multiplanar spoiled GRASS (FMPSPGR) (TR130/TE8.4-9.5). Contrast-to-noise ratios (CNRs) of HCCs determined during the imaging sequences formed the basis of quantitative analysis, and images were qualitatively assessed in terms of lesion conspicuity and image artifacts. The diagnostic accuracy of all sequences was assessed using receiver operating characteristic (ROC) analysis. RESULTS: Quantitative analysis revealed that the CNRs of T2*-weighted FMPGR and T2* weighted FMPSPGR were significantly higher than those of the other sequences, while qualitative analysis showed that image artifacts were prominent at T2* weighted GRASS imaging. Lesion conspicuity was statistically significantly less clear at SSFSE imaging. In term of lesion detection, T2*-weighted FMPGR, T2*- weighted FMPSPGR, and proton density FSE imaging were statistically superior to the others. CONCLUSION: T2*-weighted FMPGR, T2*- weighted FMPSPGR, and proton density FSE appear to be the optimal pulse sequences for ferumoxidesenhanced MR imaging in the detection of HCCs. PMID- 12087199 TI - The induction of hyperthermia in rabbit liver by means of duplex stainless steel thermoseeds. AB - OBJECTIVE: To determine the heating characteristics of needle-shaped duplex stainless steel thermoseeds, and to evaluate their effectiveness in the induction of hyperthermia in rabbit liver. MATERIALS AND METHODS: Thermoseeds of the two different shapes, L-shaped for single doses of hyperthermia and I-shaped for in vitro study and repeated hyperthermic induction, were prepared. For the in-vitro study, an I-shaped thermoseed 0.23 mm in diameter and 25 mm long was placed inside a plastic tube filled with water. Heat was applied for 30 minutes within an induction magnetic field, and during this time changes in temperature were recorded using three thermocouples. For the in-vivo study, fifteen New Zealand white rabbits were divided into five equal groups. An I-shaped or L-shaped thermoseed was inserted in each rabbit's liver, and then placed within the center of the magnetic induction coil during a 30-minute period of hyperthermia. The rabbits in the first group were sacrificed immediately after hyperthermia was induced once, while those in the other groups were sacrificed at 1, 3, and 7 days, respectively, also after one induction. The remaining three rabbits were sacrificed 4 days after three consecutive daily treatment sessions. The resected segments of liver were subsequently evaluated histopathologically for the extent of coagulation necrosis caused by heating of the thermoseed. RESULTS: The in vitro study demonstrated that the temperature in the thermoseed, which was 25.9 degrees C before heating and 54.8 degrees C after heating, rose rapidly at first but progressively less rapidly as time elapsed. Light microscopic examination of the rabbits' livers revealed coagulation necrosis and infiltration by inflammatory cells around the insertion site of the thermoseed. The maximum diameter of coagulation necrosis was 2.81+/-1.68 mm, and this occurred in the rabbits that were sacrificed 7 days after heat induction. CONCLUSION: Needle shaped duplex stainless steel thermoseeds show temperature-dependent-type heating characteristics, and in rabbit liver, induced coagulation necrosis of surrounding tissues after heat is applied for 30 minutes. These thermoseeds may thus be useful for the induction of interstitial hyperthermia. PMID- 12087200 TI - In-vivo proton magnetic resonance spectroscopy in adnexal lesions. AB - OBJECTIVE: To explore the in-vivo 1H- MR spectral features of adnexal lesions and to characterize the spectral patterns of various pathologic entities. MATERIALS AND METHODS: Thirty-one patients with surgically and histopathologically confirmed adnexal lesions underwent short echo-time STEAM (stimulated echo acquisition method) 1H- MR spectroscopy, and the results obtained were analysed. RESULTS: The methylene present in fatty acid chains gave rise to a lipid peak of 1.3 ppm in the 1H- MR spectra of most malignant tumors and benign teratomas. This same peak was not observed, however, in the spectra of benign ovarian epithelial tumors: in a number of these, a peak of 5.2 ppm, due to the presence of the olefine group (-CH=CH-) was noted. The ratios of lipid peak at 1.3 ppm to water peak (lipid/water ratios) varied between disease groups, and in some benign teratomas was characteristically high. CONCLUSION: An intense lipid peak at 1.3 ppm is observed in malignant ovarian tumors but not in benign epithelial tumors. 1H- MRS may therefore be helpful in the differential diagnosis of adnexal lesions. PMID- 12087201 TI - Fetal musculoskeletal malformations with a poor outcome: ultrasonographic, pathologic, and radiographic findings. AB - The early and accurate antenatal diagnosis of fetal musculoskeletal malfomations with a poor outcome has important implications for the management of a pregnancy. Careful ultrasonographic examination of a fetus helps detect such anomalies, and a number of characteristic features may suggest possible differential diagnoses. During the last five years, we have encountered 39 cases of such anomalies, and the typical prenatal ultrasonographic and pathologic findings of a number of those are described in this article. PMID- 12087203 TI - Mediastinal lymphangioma and chylothorax: thoracic involvement of Gorham's disease. AB - We report a case of mediastinal lymphangioma associated with Gorham's disease in a 38-year-old man who had suffered recurrent clavicular fractures during a seven year period. Mediastinal widening associated with osteolysis of the clavicles and the sternal manubrium was revealed by chest radiography, while computed tomography demonstrated a cystic anterior mediastinal mass infiltrating mediastinal fat and associated with osseous destruction of the clavicles and manubrium. Chylothorax recurred during the course of the disease. PMID- 12087202 TI - Clear-cell meningioma: CT and MR imaging findings in two cases involving the spinal canal and cerebellopontine angle. AB - Clear-cell meningioma is a rare subtype of meningioma which occurs at a younger age and has a higher recurrence rate than other subtypes. We report two cases of clear-cell meningioma, one in the thoracolumbar spinal canal and the other in the cerebellopontine angle. Though the CT and MR imaging findings were not different from those of ordinary meningioma, after surgical removal the condition recurred repeatedly in the patient with spinal canal involvement. PMID- 12087204 TI - Radiologic findings of multiple myeloma with gastric involvement: a case report. AB - We report a case of multiple myeloma with gastric involvement occurring in a patient who underwent an upper gastrointestinal series (UGIS), CT and MRI. UGIS depicted a luminal protruding mass, while contrast-enhanced CT demonstrated marked thickening of the gastric wall, with subtle contrast enhancement. At T1- and T2-weighted MR imaging, the mass showed iso- and intermediate signal intensity, respectively. After the administration of contrast material, subtle homogeneous enhancement was apparent. PMID- 12087205 TI - Imaging findings of Castleman's disease localized in the axilla: a case report. AB - Castleman's disease is a rare benign lymphoproliferative disorder of uncertain origin which most commonly involves the mediastinum but rarely affects the axilla. We report a case of localized Castleman's disease involving the axillary lymph node. Mammography revealed a well-defined, homogeneously dense ovoid mass, 3 cm in size, in the left axilla, while gray-scale ultrasonography (US) demonstrated a well-defined, uniformly hypoechoic ovoid mass with good through transmission. Peripheral hypervascularity was observed at power Dopper US, and early rapid homogeneous enhancement at contrast-enhanced dynamic CT. PMID- 12087206 TI - Fetal pericallosal lipoma: US and MR findings. AB - We report a case of fetal pericallosal lipoma occurring at the anterior interhemispheric fissure and associated with agenesis of the corpus callosum. During targeted prenatal ultrasonography at 26 weeks' gestation, the lesion was seen as a highly echogenic mass. MR imaging performed at 35 weeks' gestation and during the postnatal period revealed a pericallosal fatty mass and agenesis of the corpus callosum. PMID- 12087207 TI - [Role of parenteral cephalosporins for outpatients treatment of infections]. AB - OPAT (Outpatient Parenteral Antibiotic Therapy) arose in the early 1980s in the USA and later in many other countries from the primary consideration that outpatient treatment is more cost-effective than hospitalisation. Currently, several thousand patients undergo OPAT programmes all over the world and several different bacterial infections are included in the list of treatable diseases, especially those requiring long-term parenteral treatment such as osteomyelitis and soft tissue infections. All injectable antibiotics are suitable for OPAT according to their microbiological spectrum, although clearly some pharmacological properties make one antibiotic more preferable than another. Beta lactams represent more than half of the antibiotic world market and two-thirds of them are cephalosporins. Such a widespread use of cephalosporins is certainly due to their wide antibacterial spectrum and good tolerability. Among third generation cephalosporins, covering the majority of micro-organisms responsible for community-acquired infections, ceftriaxone is the only one with an 8-hour half-life, thereby permitting a single daily dose, which represents a great advantage when undertaking an OPAT programme. Analysis of antibiotic consumption used for OPAT therapies, based on data collected from the International OPAT Registry project, with the participation of many countries (USA, Canada, Britain, Argentina, etc.) including Italy, shows that ceftriaxone is the most widely used antibiotic for home therapy, clearly due to the above-mentioned properties. PMID- 12087208 TI - [Microbial circulation and control in two Intensive Care Units]. AB - A microbiological survey was carried out in two medical Intensive Care Units from January to June 2000. The patients, staff (hands and upper respiratory tract) and environment were monitored. The results obtained in both Care Units give cause for concern. They showed particularly high cultural positivities in bronchoaspirates collected from artificially ventilated patients, a high percentage of positive environmental samples, and frequently contaminated hands in hospital staff, conditions which may facilitate microbial circulation in the medical Intensive Care Units. It would therefore seem necessary to promptly apply specific preventive measures for both the environment and patients. PMID- 12087209 TI - [Clarithromycin switch therapy in adult patients suffering from community acquired lower respiratory tract infections (LRTIs) requiring hospitalisation]. AB - 39 subjects aged 53-85 years suffering from COPD, admitted to our Unit, were enrolled in our study: four patients were diagnosed as having CAP; 35 patients meet the criteria of COPD exacerbation. The enrolled patients had been receiving clarithromycin (CLA) 500 mg i.v. every 12 hours; switching to oral therapy was decided on the basis of Ramirez's criteria. The mean total duration (SD) of treatment with CLA was 9.56 (1.87) days. During the study, 2 patients dropped out because of the detection of malignant tumoral cells in the sputum in one case and admission to the Intensive Care Unit because of pulmonary thromboembolism in the other. The clinical assessment made 24-72 hours after the end of the treatment confirmed the clinical efficacy of CLA (cure + improvement) in 35 out of 37 evaluable patients (94.59%). At the first follow-up visit, clinical efficacy was confirmed in 34 patients (91.89%). At the long-term follow-up visit, performed 30 45 days after the end of the study, none of the 34 "cured" patients had relapsed. Local and systemic tolerability was excellent in all patients, and no side effect attributable to the drug was reported. Our experience with switched clarithromycin therapy in patients with LRTIs confirms the validity of this approach. PMID- 12087211 TI - [Infective endocarditis: a retrospective analysis of inpatient cases]. AB - The authors analysed retrospectively the clinical and laboratory features of patients hospitalised in a unit of infectious diseases in the period from 1996 99. In particular, the etiology, the antimicrobial susceptibility of responsible microorganisms, criteria utilized for diagnosis, antibiotic courses and outcomes were evaluated and discussed. Echocardiography is an essential tool in the diagnosis. Infective endocarditis remains a serious infection and is still associated with high morbidity and mortality rate, despite appropriate treatment. Seriously ill patients may require prompt surgery. PMID- 12087210 TI - [Immigration and sexually transmitted diseases: risk factors, prevention and health education]. AB - Assessment of behaviour at risk of HIV-infection and other Sexually Transmitted Diseases (STD) in high-risk subjects, such as illegal immigrants is the first step for successful prevention measures. In order to assess knowledge of AIDS, STDs, risk behaviour and condom use, a sexual behaviour questionnaire was administered to all illegal immigrants living in the Domitia area (north-west of naples) and coming to our ambulatory for several pathologies. The following risk markers identified were: low level of knowledge concerning HIV and STD transmission and prevention, multiple sexual partners, casual sex, low frequencies of condom use, drugs and alcohol use. Therefore prevention campaigns should include educational activities concerning AIDS and STD transmission and prevention, and condom promotion. Particular attention should be given to improve access to STD services that provide treatment and counselling. Moreover, commercial sex workers require counselling at each visit, screening and treatment. PMID- 12087212 TI - [Methicillin-resistant Staphylococcus aureus (MRSA) in an intensive care unit: a one-year survey]. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is frequently isolated in nosocomial outbreaks. In our study, we analysed the occurrence of colonisation and infection in an Intensive Care Unit of our hospital during a 12-month period. We also evaluated the possibility of using automated ribotyping as a molecular method in order to type the isolates. Twice a week a nasal swab and a rectal swab were performed on all patients; from ventilator-assisted patients, a sputum culture was also taken. All the MRSA isolated were identified by using commonly phenotypic procedures and on all isolates susceptibility tests were performed. An automated ribotyping using EcoRI was also done. Out of 292 patients enrolled in the study, 205 were never colonised (group N); among the other 87 who were colonised by MRSA (29.8%), 40 patients (group A) were MRSA carriers at the time of admission, while 47 (group B) were colonised in the ICU. Twenty-seven patients (11 from group A, 15 from group B and 1 from group N) developed 31 infections due to MRSA. Patients from group A exhibited, as a rule, worse clinical conditions than those from the other two groups. For the former group, MRSA infection was frequently systemic (sepsis), while in group B pneumonia was the predominant infection. The prevalence of colonisations in our study was 30%, which is a value comparable to those presented by other authors in similar cases. MRSA colonisation is a necessary condition for subsequent infections in almost all cases, with an average lag of 7 days. Susceptibility tests were non discriminating among the isolates: all the strains were susceptible to glycopeptides; nearly all of them were resistant to erythromycin, clindamycin, ciprofloxacin and gentamicin. Automated ribotyping allowed us to distinguish 12 different ribogroups, the most frequent of which was composed of 146 isolates. In our study, this molecular method was able to define a possible endemic clone that should be better investigated by using methods with a higher discriminatory power, such as RAPD or PFGE. The method that we employed is highly reliable, easy to perform and not time-consuming. In our opinion, it could be the method of choice in the first screening of high numbers of isolates. PMID- 12087213 TI - [Tuberculous meningitis in childhood: review of cases observed in the years 1995 1999]. AB - In this study we reviewed M. tuberculosis meningitis (TBM) cases in childhood at our unit in a five-year period. METHODS: Included in this retrospective study were children admitted to Cotugno Hospital from 1995 to 1999 who fulfilled criteria for a TBM diagnosis. Medical records were analyzed for epidemiological data, clinical manifestations, laboratory test results, radiological investigations and clinical outcome. RESULTS: We identified 10 children with TBM. Family history of tuberculosis (TB) was observed in 6 patients. Fever preceded hospital admission of 4-60 days (median 8 days); neurological signs were present 1-6 days before admission in 8 patients. All patients showed a predominance of lymphocytes, elevated proteins and reduced glucose concentration in cerebrospinal fluid (CSF). Mycobacterium tuberculosis was either cultured or identified by acid fast stain in 5 patients. Cranial CT-scans and MRI showed hydrocephalus in 8 patients and ring-enhancing lesions in 6. Antituberculous therapy was administered according to current recommendations for at least 12 months, with a 4-6 week steroid therapy course. Two patients had permanent neurological sequelae. CONCLUSIONS: Careful evaluation of symptoms and CSF findings is the only way to establish an early diagnosis and reduce sequelae. PMID- 12087214 TI - [Relationship between malaria, environment, people and civilisation in central Italy. Reclamation of the Fucino plain]. AB - In this article the incidence of malaria in the course of history is cited as a paradigmatic example of pathocenosis - emergence of new infectious diseases related to the environmental conditions or to the different ecology of the infected host - as malaria is affected by climatic variations and environmental deterioration. The incidence of malaria in the Mesolithic and Neolithic ages is briefly evaluated. Moreover, the finds from a necropolis in the Sele river plain established the effect that the land reclamation of the wetland during the Etruscan period induced on the porotic hyperostosis, an indirect index of malaria sustained by Plasmodium falciparum. In addition, we consider the reclamation in the area of the Fucino plain, a volcanic basin without outflow, whose shores were therefore unhealthy. In the reign of the Claudius, in the early years of the Roman Empire, a tunnel was built to provide an outflow, thereby draining the wetlands. The results of this engineering intervention lasted for six centuries. During medieval times the area was neglected, thus creating the pre-existing insalubrious environmental conditions. It was only during the Bourbon period in the 19th century that a new intervention was initiated, subsequently completed by Prince Torlonia, which allowed the wetlands to be definitively transformed into farmland. PMID- 12087215 TI - [Sexually transmitted diseases (STD) and their relationship with sexual behaviour and condom use, in a cohort of teenagers referring to a STD centre. A nine-year, prospective study]. AB - In order to assess the relationship between a diagnosis of sexually-transmitted disease (STD), sexual behaviour, condom use, and other social, demographic, and epidemiological variables in a cohort of young adults who referred to a STD centre during a 9-year period, all patients aged 13-20 years were prospectively evaluated, with special attention paid to sexual behaviour, and use of condom or other contraceptive techniques. The 284 assessed young adults represented 6.1% of all patients with a diagnosis of STD: an increasing temporal trend was noticed (from 3. % in 1991, up to 10.4% nel 1999: p<.0001). On the whole, 70.1% of subjects aged 20 years or less never used a condom during the 6 months preceding the diagnosis of STD: only 21.8% of patients reported regular condom use, and 4.9% more subjects referred occasional use, while in the remaining 3.2% of cases other contraceptive methods were employed. Among under age patients (10.6% of study population), the rate of condom use was 3.3% only, while male homosexuals always denied the use of barrier methods. Although a progressive increase in condom use was observed through the study time (from 0% encountered in 1991-1994, to 57.6% of 1999: p<.0001), this phenomenon proved linked only to the massive increase of immigrant sex workers, which occurred since 1997. When excluding from analysis any probable female prostitutes, lack of condom use tested related to the male gender (p<.05), and a low education (p<.04). Over 50% of patients reported 0-1 partners during the last 6 months, even though an increase in sexual promiscuity was observed during the last three years, compared with 1991-1996 (p<.02). Among the 297 different STD episodes (mostly nongonococcal cervicitis vaginitis and urethritis, and HPV infection), an increasing incidence of nongonococcal STD and syphilis was found, but no correlation was observed between the spectrum of diagnosed STD, sexual behaviour, and condom use. In particular, in 69.9% of the 103 episodes nongonococcal disease occurred despite regular (65 cases), or occasional (7 episodes) condom use. Moreover, no significant relationship was disclosed between STD occurrence, condom use, and other analyzed social, demographic, epidemiological, and clinical variables. According to our survey, an increased risk of STD seems to involve young adults, and to be related to the variation of multiple demographic, epidemiological, and behavioral features. While immigration and prostitution had the major impact during the last three years, sexual promiscuity and infrequent condom use represent persistent; risk factors in this age population. Besides their diagnostic and therapeutic role, STD centres are able to carry out permanent monitoring of STD, as a starting point to plan adequate information campaigns and specific prevention strategies. PMID- 12087216 TI - [Epidemiological features of intestinal parasitosis in western Sicily in the period 1993-2000]. AB - To determine the prevalence of intestinal parasitic infections in western Sicily from 1993 to 2000, fecal samples from 3024 patients (subdivided into native Sicilian children, migrants from developing countries and HIV+ patients,) were collected and observed on direct examination or by the formalin-ether sedimentation technique of Ritchie. Lugol, Kinyoun and Gomori's trichromic stain techniques were employed. Trophozoites or mainly protozoan cysts and helminthic eggs were detected in 195 patients (6.4%). The positivity rate was 18.7% in HIV+ patients, 10.3% in migrants and 5.0% in native Sicilian children. Tape test for Enterobius eggs was employed on 1449 patients with a positivity rate of 11%. The importance of parasitology also in new millennium is emphasized. PMID- 12087217 TI - [Complications of mediterranean spotted fever]. AB - Mediterranean spotted fever is an infectious disease due to Rickettsia conori transmitted to man by the dog tick Rhipicephalus sanguineus. The review of a consecutive series of 525 cases, admitted to Caltagirone hospital in the last 20 years, permits the classic clinical picture to be identified by fever, maculopapular eruption and tache noire, and any complications to be visualized. Usually the course of the disease is good, but 12.7% of our cases reported complications such as renal failure, myocarditis, pneumonia, encephalitis, anicteric hepatitis, gastrointestinal bleeding, anaemia and impaired glucose tolerance. The development of a systemic vasculite is the main pathogenetic factor in the origin of systemic complications. Early diagnosis and specific antibiotic treatment may reduce the risk of complications. PMID- 12087218 TI - [Evaluation of the efficacy of a program to control nosocomial spread of methicillin-resistant Staphylococcus aureus]. AB - OBJECTIVE: To evaluate the efficacy of a program to control nosocomial spread of methicillin-resistant Staphylococcus aureus (MRSA). METHODS: Analysis of the incidence of infection and contamination due to MRSA in patients admitted to the hospital of Cremona 6 months before and 3 years after the introduction of the guidelines (July 1997). RESULTS: During the 42 months of the study period, on 80705 admissions, 511 cases of MRSA contamination/infection were identified, the incidence being 0.57 cases per 100 admissions. The infection rate dropped from 0.34 (IC95%: 0.25-0.45) in the first 6 months of the study, before the introduction of guidelines, to 0.17 (IC95%: 0.14-0.20) in the following 3 years (p=0.01). Severe infection decreased from 0.18 to 0.1 per 100 admissions, with a 44% decrease (p=0.058), while mild infections diminished from 0.16 to 0.07 per 100 admissions (p=0.045). Methicillin resistance among nosocomial isolates of Staphylococcus aureus was reduced from 53 % to 35 % (p<0.0001). CONCLUSIONS: The introduction of a program to control the nosocomial spread of MRSA proved effective in reducing both the incidence of infection and the methicillin resistance of Staphylococcus aureus isolates. The cost effectiveness of the program seems very favourable. PMID- 12087219 TI - [Experience at a branch center of the National HIV Protease Inhibitor Registry]. AB - In the present study we show our experience as one of the Centers (center 006) of the Italian Index of Protease Inhibitors (PI). In this multicentric, observational study we studied HIV-positive naive patients for antiretroviral therapy with protease inhibitors. From 8/1/97 to 31/1/98 in our department 64 patients were enrolled and followed up through an electronic medical sheet in order to evaluate efficacy, tolerability, toxicity and compliance of treatment with PI. Our results show a reduction in AIDS-related mortality and a decrease in AIDS defining diseases: in particular data show an increase in CNS pathologies in comparison with other opportunistic events. Moreover, in patients non complaints to therapy, the progression of the disease was more frequent. The side effects more frequently observed during PI treatment were hypertrigliceridaemia and lipodystrophy; gynaecomastia and hyperprolactinaemia were even present, perhaps as expression of direct metabolic action of HAART therapy. PMID- 12087220 TI - A case of flare syndrome. AB - The authors describe a case of severe CMV retinitis in a young adult AIDS patient who recovered following first a course of ganciclovir and then HAART. Six months after the initial episode while still under successful HAART, the patient developed an acute episode of retinitis despite a persistent significant improvement in the immunological picture and a very low level of CMV reactivation. The acute episode can be related to an enhanced individual reactivity to minor CMV replication. PMID- 12087221 TI - A contraindication for the use of hydroxyapatite cement in the pediatric population. AB - The authors report on their experience with a particular hydroxyapatite cement in the pediatric population and review cases that elucidate potential limitations of its use in this population. In all patients, the implant was used for recontouring and augmenting the cranial vault. Seven of 15 patients had a direct communication between sinus mucosa and the implant at the time of surgery. Three of the seven patients developed late postoperative inflammatory reactions of the surgical site with delamination of the implant. Each patient required an aggressive irrigation and debridement with removal of the delaminated hydroxyapatite. Pathologic findings showed evidence of a foreign body inflammatory reaction in the tissue, and mixed bacterial flora were identified in all specimens. No late problems with the cement were seen in patients without a communication to the sinuses. Our findings suggest that the use of BoneSource, an essentially nonviable, nonvascularized implant, in areas exposed to bacterial contamination is contraindicated in the pediatric population. BoneSource and other similar hydroxyapatite biomaterials do have a role as implants in the craniofacial skeleton. The indications and contraindications of these substances will need to be further defined for wide acceptance to occur. PMID- 12087224 TI - The soft triangle revisited. AB - Five cadaveric adult human noses were examined to further elucidate the region of the soft triangle of the nose. Three distinct zones were found. Zone 1, just caudad to the lower lateral cartilages, consists of the fibers of the dilator nares anterior as it inserts into the dermis. Zone 2 is the dermal layer. Zone 3 is at the nostril rim and consists of muscle fibers interdigitating within the dermis; laterally, these fibers are continuations of the nasalis muscle, whereas medially they are extensions of the depressor septii muscle. Perpendicular incisions through any of these zones may cause notching and tip deformity. Incisions in zone 1 may, in addition, alter external nasal valve function by interrupting the fibers of the dilator naris anterior. PMID- 12087222 TI - Postoperative mental and morphological outcome for nonsyndromic brachycephaly. AB - Bilateral coronal synostosis causes functional and morphological problems that require fronto-orbital advancement in infancy to correct the brachycephalic deformity and to prevent mental impairment caused by the intracranial hypertension. In this study, 99 children with isolated cases of brachycephaly were prospectively followed to study their preoperative and postoperative mental outcome, which was evaluated using developmental or intelligence quotients. Several factors were analyzed: age before treatment, age at the time of surgery, and the correlation between mental assessments before and after surgery. In a subgroup or patients tested for the FGFR3 P250R mutation (n = 48), mental and morphological assessments were analyzed. Before surgery, mental status was better in the patients tested before 1 year of age (p < 0.001). The preoperative mental assessment always correlated with the postoperative assessment (p < 0.0001). The postoperative mental outcome was better when surgery was performed before the patient reached 1 year of age (p < 0.02). Although both the morphological and functional outcomes were better in the subgroup of noncarriers of the mutation, the differences were not statistically significant. Prominent bulging of the temporal fossae was frequently responsible for poor morphological outcome in carriers of the mutation. This study confirms the need for early corrective surgery before 1 year of age in brachycephalic patients to prevent impairment of their mental development. Suboptimal morphological and mental outcomes can be expected in patients with nonsyndromic brachycephaly who carry the FGFR3 P250R mutation. Primary correction of the temporal bulging should be performed in conjunction with fronto-orbital advancement to improve the morphological outcome in patients with the mutation. PMID- 12087225 TI - Primary definitive nasal correction in patients presenting for late unilateral cleft lip repair. AB - Almost 25 percent of unilateral cleft lip and palate patients present with their deformity in their teens or later years in the developing world. Because more than 80 percent of the world population lives in the developing world, the established protocol for repair of these deformities is not applicable to these patients. Despite the magnitude, there are no significant reports in the literature that deal with this problem. Several issues need to be addressed, but the author limits himself here to the correction of the nasal deformity. The patients at this age are very much concerned with the aesthetic outcome. Procedures described hitherto for primary nasal correction in infants are not successful in restoring nasal shape and symmetry at this late age of presentation. Our experience with radical correction of secondary nasal deformity in unilateral cleft lip patients presenting late prompted us to extend the concept by undertaking a definitive primary correction of the nasal deformity in cleft patients presenting late. Twenty-two patients with unilateral cleft lip deformity (nine male patients and 13 female patients) with ages ranging from 13 to 22 years, presenting between August of 1997 and December of 2000, are included in this study. Of these, 11 patients had a cleft of the lip alone, eight also had a cleft of the alveolus, and three had a cleft of the palate continuous with the cleft lip. All patients showed some maxillary hypoplasia. An external rhinoplasty with lip repair was carried out in all patients. The corrective procedures on the nose included columellar lengthening; augmentation along the pyriform margin, nasal floor, and alveolus using bone grafts; submucous resection of the nasal septum; repositioning of lower lateral cartilages; and augmentation of nasal dorsum by bone graft. Clinical follow-up ranged from 4 to 24 months, and the median follow-up period was 13 months. Results have been very good, and much better than results seen earlier with other primary rhinoplasty techniques. While repairing unilateral cleft lip in adolescents, the author thinks it would be most appropriate to address the entire gamut of the deformity in a single stage, provide complete vector reorientation, and augment the hypoplastic elements by autologous tissue. It is not just the fear of poor follow-up, but that merely correcting the lip deformity in these patients without attempting definitive rhinoplasty, in the author's opinion, would be insufficient surgical intervention. PMID- 12087226 TI - Treatment of mandible fractures using bioabsorbable plates. AB - This study evaluated the short-term outcome of treating fractures of the mandible with bioabsorbable plates. Sixty-nine fractures of the mandible in 49 patients were treated by open reduction and internal fixation using bioabsorbable plates and 2.4-mm, 2.0-mm, and 1.5-mm pretapped screws. The duration of intermaxillary fixation ranged from 0 to 23 days, with a mean of 4.6 days. Patients were evaluated for complications during the follow-up period, which ranged from 1 to 18 months. Six patients (12.2 percent) experienced complications. These included infection (four patients), premature occlusal contact (one patient), and temporomandibular disorder (one patient). Except for one case, all complications were minor and adequately managed with incision and drainage, elastic traction, physical therapy, and medication. Delayed infection (osteomyelitis) developed in a symphysis fracture and was treated by saucerization and antibiotics. The fracture line subsequently showed complete consolidation. Bioabsorbable plates can be selectively used for internal fixation in mandibular fractures, with the advantage that they do not need to be removed. PMID- 12087229 TI - Use of free thin anterolateral thigh flaps combined with cervicoplasty for reconstruction of postburn anterior cervical contractures. AB - Free thin anterolateral thigh flaps combined with cervicoplasty were used in a series of seven patients undergoing reconstruction for previous burn injury from September of 2000 to May of 2001 at Chang Gung Memorial Hospital. This method uses a suprafascial dissection technique to provide a thin flap to improve cervical contour. Neck contractures had resulted from flame burns in six patients and from a chemical burn in one patient. The mean age was 32.7 years (range, 22 to 45 years). The size of excised scar ranged from 10 x 2 cm to 26 x 5 cm (mean, 19.7 x 3.3 cm). The size of flaps ranged from 11 x 5 cm to 26 x 8 cm (mean, 21.3 x 6.5 cm). Average operative time was 6 hours. Average hospital stay was 10 days. All flaps survived, with one flap sustaining partial marginal loss. The donor site was closed primarily in five cases and by using a split-thickness skin graft in two cases. At a mean follow-up time of 5 months, the functional improvement was measured as follows: a mean increase in extension of 30 degrees (preoperatively, 95 degrees; postoperatively, 125 degrees), a mean increase in rotation of 18 degrees (preoperatively, 59 degrees; postoperatively, 77 degrees), and a mean increase in lateral flexion of 12.5 degrees (preoperatively, 26.5 degrees; postoperatively, 39 degrees). The average cervicomandibular angle was improved by 25 degrees (preoperatively, 145 degrees; postoperatively, 120 degrees). This series demonstrates that the use of free thin anterolateral thigh flaps combined with cervicoplasty provides a one-stage reconstruction with a thin, pliable flap that achieves good cervical contour with low donor-site morbidity. PMID- 12087228 TI - Squamous cell carcinoma of the oral mucosa after release of submucous fibrosis and bilateral small radial forearm flap reconstruction. AB - Oral submucous fibrosis is a collagen disorder that affects the submucosal layer of the upper digestive tract. The major cause is the habit of betel quid chewing, which is common in central, southern, and southeast Asia. The progressive and irreversible course of disease results with trismus, dysphagia, xerostomia, and rhinolalia. The most serious complication of this disorder is the development of oral carcinoma, and the incidence in different series varies from 1.9 to 10 percent. A sufficient mouth opening can be achieved by complete release of fibrotic tissue, and coronoidectomy and temporal muscle myotomy when needed, and reconstruction of the resultant defect can be best achieved by microsurgical free tissue transfer because of the discouraging results with skin grafting or local flaps. From April of 1997 to May of 2001, a total of 26 patients received reconstructive surgery with small radial forearm flaps after release of submucous fibrosis with or without temporalis muscle myotomy and coronoidectomy. All patients were men, with a mean age of 40.1 years (range, 18 to 62 years) and all had a history of betel nut chewing ranging from 8 to 40 years. The interincisal distance ranged from 5 to 29 mm, with a mean of 15 mm, before operation. After the release procedure, the interincisal distance increased to 40 mm (range, 35 to 50 mm). At a follow-up period of 3 to 48 months, the interincisal distance was a mean of 35 mm (range, 18 to 57 mm), with an average increase of 20 mm compared with the preoperative distance. During follow-up, three patients developed squamous cell carcinoma of the oral cavity 24 to 36 months after submucous fibrosis release. Two of them occurred in the release site and the other one occurred at the soft palate. Oral cancer occurred in three of 13 patients who had received release of submucous fibrosis and who were followed for longer than 2 years (range, 24 to 48 months), which means that 23 percent of these patients developed squamous cell carcinoma of the intraoral mucosa. High risk of cancer occurrence strongly indicates the importance of an earlier and more aggressive surgical approach toward submucous fibrosis, and long-term follow-up on a regular basis. The purpose of an early and aggressive approach to submucous fibrosis is to provide a good quality of life to the patient by improving oral hygiene and oral intake quality and at the same time to obtain a sufficient mouth opening, which is mandatory for the inspection of the excision site and the remaining oral mucosa during follow-up. PMID- 12087230 TI - A newly defined conchal floor arterial flap for auricular reconstruction. AB - A conchal floor composite flap pedicled by skin of the helical crus has been well described in the literature. Here the flap is elevated based on the supply by an upper auricular branch of the superficial temporal artery. In this article, a newly defined conchal floor arterial flap is proposed. The flap is based on the main stem of the posterior auricular artery and its venae comitantes. Two types of conchal floor arterial flaps were elevated: a proximally based chondral arterial flap and a distally based chondrocutaneous arterial flap. The proximally based flap was used for earlobe reconstruction, whereas the distally based flap was useful in the reconstruction of the upper auricle. Nine congenital auricular malformations were successfully corrected with this newly defined conchal floor flap procedure. This type of flap is easier to elevate, more reliable, and more versatile than the one currently in use. PMID- 12087231 TI - Polytetrafluoroethylene bars in stabilizing the reconstructed sternum for pectus excavatum operations in children. AB - Nine children with severe pectus excavatum deformity were operated on in the Department of Pediatric Surgery, Thoracic Unit, Ege University Faculty of Medicine, Izmir, Turkey. There were four boys and five girls, and the mean age at operation was 6.3 years. A modified Ravitch technique was performed by using two polytetrafluoroethylene bars placed retrosternally for fixation and stabilization of the reconstituted anterior chest wall. The bars were removed 6 months after the operation. Results were satisfactory in all cases, with no surgical complications. PMID- 12087232 TI - Clinical applications of three-dimensional photography in breast surgery. AB - Three-dimensional imaging in breast surgery has several uses clinically. The most practical applications are for the evaluation of breast asymmetries, both congenital and acquired, and for the evaluation of factors affecting breast shape in augmentation mammaplasty. Other uses of three-dimensional imaging that we have found clinically helpful are for evaluation of patients desiring reduction mammaplasty and for evaluation of patients undergoing unilateral breast reconstruction to determine the expander and permanent implant size that gives the best symmetry with the contralateral breast. We present five cases in which we investigate the use of three-dimensional imaging clinically by using the images to determine quantitative information about the breast, such as volume or projection. Overall, three-dimensional imaging is very helpful in providing objective information about the breast for use in preoperative planning. In addition, by analyzing clinical cases, it can provide objective data about the breast and surgical mammaplasty (especially augmentation mammaplasty) that may help surgeons better understand those factors that contribute to breast shape and influence surgical outcomes. There are currently some limitations of this system, influenced by patients with significant ptosis or obesity, which may introduce errors into the three-dimensional data, making them unreliable. However, we believe three-dimensional imaging has great clinical potential in surgical mammaplasty. PMID- 12087233 TI - Outcome analysis of reduction mammaplasty using the modified Robertson technique. AB - The benefits of reduction mammaplasty have been well documented in previous literature. Anticipating and correcting for pseudoptosis (bottoming-out), however, can impair the cosmetic outcome as the inferior skin envelope stretches and lengthens over time. We present long-term results on patients using the modified Robertson technique for reduction mammaplasty, which appears to have significant benefit in helping to prevent bottoming-out. Surveys were sent to patients undergoing reduction mammaplasty surgery with this technique from 1987 to 1997. Patients were queried regarding preoperative and postoperative symptoms, satisfaction, and outcome related to their surgery and were also offered free follow-up examinations. The patients who returned for follow-up were then evaluated by the attending surgeons for evaluation of scarring, nipple position, ptosis, pseudoptosis, shape, and overall appearance. Reduced breasts were also compared with cosmetically optimal breasts to compare for measured levels of pseudoptosis using our defined visual inferior pole ratio measurements. Average reduction size was 910 g and follow-up was 4.7 years from the time of surgery. There was significant improvement demonstrated in all areas questioned, with the greatest relief shown in back and shoulder pain, shoulder grooving, and difficulty fitting clothing. There was also demonstrated to be significantly less use of medical modalities postoperatively and significant increases in activity levels. Satisfaction for size, shape, symmetry, and overall results was 85, 94, 98, and 94 percent, respectively. Evaluations for pseudoptosis by the attending surgeons were rated good or excellent in 95 percent of patients. Measurements of the visual inferior pole ratio for pseudoptosis also demonstrated no significant differences when compared with aesthetically optimal breasts. The modified Robertson reduction mammaplasty is a reliable technique that can be used for both small and large reductions, giving both reliable and consistent results. This technique significantly improves symptoms, as do other reduction techniques, but has the added advantage of helping to avoid pseudoptosis postoperatively. PMID- 12087235 TI - Anterolateral thigh flap for postmastectomy breast reconstruction. AB - Most postmastectomy defects are reconstructed by use of lower abdominal-wall tissue either as a pedicled or free flap. However, there are some contraindications for using lower abdominal flaps in breast reconstruction, such as inadequate soft-tissue volume, previous abdominoplasty, lower paramedian or multiple abdominal scars, and plans for future pregnancy. In such situations, a gluteal flap has often been the second choice. However, the quality of the adipose tissue of gluteal flaps is inferior to that of lower abdominal flaps, the pedicle is short, and a two-team approach is not possible because creation of the gluteal flap requires that the patient's position be changed during the operation. In 2000, five cases of breast reconstructions were performed with anterolateral thigh flaps in the authors' institution. Two of them were secondary and three were immediate unilateral breast reconstructions. The mean weight of the specimen removed was 350 g in the three patients who underwent immediate reconstruction, and the mean weight of the entire anterolateral thigh flap was 410 g. Skin islands ranged in size from 4 x 8 cm to 7 x 22 cm, with the underlying fat pad ranging in size from 10 x 12 cm to 14 x 22 cm. The mean pedicle length was 11 cm (range, 7 to 15 cm). All flaps were completely successful, except for one that involved some fat necrosis. The quality of the skin and underlying fat and the pliability of the anterolateral thigh flap are much superior to those of gluteal flaps and are similar to those of lower abdominal flaps. In thin patients, more subcutaneous fat can be harvested by extending the flap under the skin. Use of a thigh flap allows a two-team approach with the patient in a supine position, and no change of patient position is required during the operation. However, the position of the scar may not be acceptable to some patients. Therefore, when an abdominal flap is unavailable or contraindicated, the creation of an anterolateral thigh flap for primary and secondary breast reconstruction is an alternative to the use of lower abdominal and gluteal tissues. PMID- 12087236 TI - Trends in unilateral breast reconstruction and management of the contralateral breast: the Emory experience. AB - Recent trends in breast reconstruction have transitioned toward the skin-sparing type of mastectomy and immediate reconstruction using autologous tissue. This study was designed to document trends in the management of patients with unilateral breast cancer and to determine how they influence management of the contralateral breast. All patients who underwent unilateral breast reconstruction at Emory University Hospitals from January of 1975 to December of 1999 were reviewed. The cohort was stratified by timing of reconstruction (immediate versus delayed), method of reconstruction, and mastectomy type (skin-sparing versus non skin-sparing). The methods of reconstruction included implant, latissimus dorsi flap, and transverse rectus abdominis musculocutaneous (TRAM) flap. Contralateral procedures to achieve symmetry included augmentation, mastopexy, augmentation/mastopexy, and reduction. A total of 1394 patients were evaluated, including 689 delayed and 705 immediate reconstructions. Sixty-seven percent of delayed-reconstruction patients (462 of 689) had a symmetry procedure performed on the opposite breast, compared with 22 percent for the immediate-reconstruction patients (155 of 705) (p 0.05). Neuromuscular blockade (NMB) was used in 113/180 (63%) of preAEP intubations and 145/192 (76%) of postAEP intubations (P < 0.01). NMB without sedation decreased from 62/113 (55%) in the preAEP group to 12/145 (8%) in the postAEP group (P < 0.001). Cricothyrotomy rate decreased from 10/180 (6%) in the preAEP group to 3/192 (2%) in the postAEP group (P < 0.05). Failed intubation rate in nonarrested patients during the 6-year period was 10/154 (6%) in patients receiving no medications or partial rapid sequence intubation (RSI) compared with 3/184 (2%) in patients who had full RSI (P < 0.025). CONCLUSION: Establishment of an AEP resulted in an increase in NMB for intubation, a dramatic decrease in the use of NMB without sedation, and a decrease in cricothyrotomy rate. The rate of intubation failure in nonarrested patients was higher in those who received no medications or partial RSI compared with full RSI. PMID- 12087322 TI - Etomidate as a sole agent for endotracheal intubation in the prehospital air medical setting. AB - INTRODUCTION: Etomidate is an anesthetic agent with rapid onset, short duration of action, a generally stable hemodynamic profile, and cerebroprotective effects. It is used in the hospital setting to facilitate emergency endotracheal intubation. This helicopter EMS used etomidate as an intubating agent without paralytics for 2 years. METHODS: Intubations performed by the helicopter crew using etomidate alone were reviewed. Intubation was classified as successful or unsuccessful. Successful intubations requiring three or more attempts or repeated doses of etomidate were interpreted as difficult. RESULTS: Fifty patients received etomidate to facilitate orotracheal intubation. Etomidate was the sole agent in 44 of these cases. Mean age was 31 years (range 4-79); 35 patients (80%) were men. Most patients (79%) were victims of blunt trauma. The mean dose of etomidate was 0.5 mg/kg (range 0.3-1.1). Hemodynamic parameters remained stable. Intubation was successful in 39 patients (89%). Intubation was difficult in seven patients (16%) and unsuccessful in five (11%). Masseter muscle spasm was noted in three of the five patients for whom intubation was unsuccessful. Other complications included emesis in eight patients and seizure-like activity in one patient. CONCLUSION: Etomidate can be used to facilitate emergency endotracheal intubation in the prehospital air medical setting, with a success rate of 89%. At the doses used in the study, hemodynamic parameters remained stable, but intubation was difficult or unsuccessful in 27% of patients. Masseter muscle spasm, which may represent orofacial myoclonus or inadequate relaxation, is common in patients who cannot be intubated with etomidate. Etomidate is recommended as a sole agent for facilitating intubation only when rapid sequence intubation with paralysis is contraindicated or otherwise undesirable. PMID- 12087323 TI - The effect of optical design on micromanipulator spot size using CO2 laser irradiation. AB - OBJECTIVE: The study goal was to compare the laser spot size created using reflective and refractive micromanipulators with a CO(2) laser and to determine the sensitivity of spot size to laser power. STUDY DESIGN AND SETTING: A CO(2) laser and operating microscope (400-mm focal length) was coupled to either a reflective (Cassegrain-like) or refractive micromanipulator. Laser spot size was determined by measuring the region of ablation created by laser irradiation of wood (dry tongue depressors), exposed photographic film, and agar gel using optical micrometry. Laser power varied from 0.5 to 20 W with pulse durations of 0.1 and 0.5 second. RESULTS: The reflective micromanipulator demonstrated overall smaller spot sizes for a given laser power and lower incremental change in spot size with increasing power. The reflective design demonstrated less sensitivity to increases in laser power. CONCLUSIONS: Micromanipulator optical design can result in significant differences in laser spot size. The reflective device used in this study demonstrated less sensitivity to increasing laser power. PMID- 12087324 TI - Expression of p63 and 14-3-3sigma in normal and hyperdifferentiated mucosa of the upper aerodigestive tract. AB - OBJECTIVE: Our goal was to analyze p63 and 14-3-3sigma expression in normal and hyperdifferentiated head and neck mucosa. STUDY DESIGN: Compare the in vivo expression of p63 and 14-3-3sigma by immunohistochemistry in normal mucosa and oral lichen planus, a benign mucosal lesion marked by hyperdifferentiation and apoptosis. RESULTS AND CONCLUSION: p63 is underexpressed and 14-3-3sigma is overexpressed in lichen planus on immunohistochemical analysis. SIGNIFICANCE: The findings support the hypothesis that p63 plays an antidifferentiation role, whereas 14-3-3sigma plays a prodifferentiation role in the upper aerodigestive tract epithelium. Lichen planus is a valuable model for the study of p63, 14-3 3sigma, and mucosal differentiation. p63 and 14-3-3sigma may be molecular markers for oral lichen planus. PMID- 12087325 TI - Induction chemotherapy followed by concomitant chemoradiation-induced regression of advanced cervical lymphadenopathy in head and neck cancer as a predictor of outcome. AB - OBJECTIVE: We sought to determine whether induction chemotherapy followed by concomitant chemoradiation (ICCR)-induced advanced neck disease regression could predict outcome, especially the need for complete neck dissection in patients with N2-3 stage IV head and neck cancer (HNC). METHODS: A retrospective study of 339 patients evaluated for treatment of stage IV HNC during the years 1988 to 1997 revealed 36 individuals with N2-3 cervical lymphadenopathy who were treated with ICCR. Responses to treatment, patterns of failure, and survival rates were analyzed. RESULTS: Primary and regional tumor regressions were complete in 21 patients (58%), partial in 9 (25%), and absent in 6 (17%); the corresponding local failure rates were 5%, 44%, and 33% (P < 0.02). The regional failure rates were 24%, 89%, and 83%, respectively (P < 0.001); distant failure rates were 10%, 0%, and 0% (P > 0.99). The estimated 2-year survival rates for complete and partial/nonresponders were 57% and 20%, respectively (P < 0.02). CONCLUSION: Patients with advanced regional metastases of HNC who respond completely to ICCR have an excellent chance for survival. However, such ICCR-induced complete regression of regional tumor cannot reliably predict ultimate neck disease control. PMID- 12087326 TI - Immune mediators in allergic rhinitis and sleep. AB - OBJECTIVE: Our study goal was to examine polysomnography, indices of sleep and allergy, and serum and nasal cytokines in allergic and nonallergic subjects. STUDY DESIGN AND SETTING: In this descriptive, exploratory study, 4 allergic and 4 nonallergic subjects underwent 2 nights of polysomnographic recording with serial measurements of cytokines and completed measures of sleep quality and allergic symptoms. RESULTS: Three serum cytokines (interleukin [IL]-1beta, IL-4, and IL-10) were higher in allergic subjects and were termed proallergic. Three serum cytokines (IL-1ra, IL-2, and IL-12) were higher in nonallergic subjects and were termed allergy inhibitory. Proallergic serum cytokines correlated with increased latency to rapid eye movement sleep, decreased time in rapid eye movement sleep, and decreased latency to sleep onset. Low levels of allergy inhibitory serum cytokines were associated with increased allergic symptoms. CONCLUSIONS: Differences in serum cytokines between allergic and nonallergic individuals are associated with variations in polysomnography and allergic symptoms. SIGNIFICANCE: Understanding these mechanisms may suggest novel approaches to alleviating drowsiness and other symptoms in allergic patients. PMID- 12087327 TI - Rhinosinusitis treatment protocol: changing provider habits in primary care. AB - OBJECTIVE: Sinus infections are common and represent a high cost to the health care industry. Clinical protocols can be used to develop cost-effective treatment strategies. Our objective was to show that a protocol for rhinosinusitis can change provider practice patterns in primary care. STUDY DESIGN AND SETTING: We used a literature search, survey of primary caregivers, and chart review from an academic health center to define and document current patterns. A protocol was developed and evaluated using the Rhinosinusitis Disability Index. RESULTS: Survey and chart review identified that primary caregivers do not elicit a detailed history for rhinosinusitis. Protocol use increased this to 100%. First line antibiotic therapy was used for only 49.4% of the patients before protocol use and for 68.5% after protocol use. Patients felt better after treatment 84.1% of the time; the Rhinosinusitis Disability Index could not measure improvement. CONCLUSION: Provider practice patterns were positively influenced by use of the protocol. SIGNIFICANCE: Clinical protocols should be developed to direct provider treatment practices. PMID- 12087328 TI - Nasal endoscopy and the definition and diagnosis of chronic rhinosinusitis. AB - OBJECTIVES: Although endoscopy has been shown by a few authors to be a valuable tool for the diagnosis of chronic rhinosinusitis, its true role in the evaluation of the patient with chronic rhinosinusitis has not been elucidated. The current definition of chronic rhinosinusitis is a symptom-based definition, and objective testing such as endoscopy or computed tomography (CT) is not included. However, the current treatment paradigm for chronic rhinosinusitis is dependent on the definition for diagnosis. Patients are treated with 4 weeks of antibiotics and decongestant/antihistamines/steroids based on the definition. This study aims to evaluate in a prospective fashion the place of endoscopy in the diagnosis of chronic rhinosinusitis. STUDY DESIGN: A group of 78 patients meeting the definition of chronic rhinosinusitis were subjected to same-day endoscopy and CT scanning. RESULTS: Seventeen (22%) of 78 patients had positive endoscopic and CT results. There were 20 (26%) of 78 patients with negative endoscopic and positive CT results. Six (8%) patients had positive endoscopic and negative CT results, and 35 (45%) had negative endoscopic and negative CT results. Overall, 37 (47%) patients had positive CT results, and 41 (53%) patients had negative CT results. Endoscopy showing the presence of purulence, nasal polyps, or watery congested mucosa correlated well with CT results. Negative endoscopy correlated with CT results in 65% of patients. CONCLUSION: The use of endoscopy to corroborate the diagnosis in nonpolypoid or nonpurulent rhinosinusitis in previously unoperated patients is questioned. Patients who meet the subjective definition of chronic rhinosinusitis should have a high degree of sensitivity and specificity with endoscopy or CT. The fact this is not the case questions the accuracy of the definition and the treatment paradigm. SIGNIFICANCE: According to this study, positive endoscopic results correlated well with CT, and negative endoscopic results correlated in 71% of patients with negative CT results. PMID- 12087329 TI - Long-term outcome of facial growth after functional endoscopic sinus surgery. AB - OBJECTIVE: We sought to determine whether functional endoscopic sinus (FES) surgery performed in children with chronic rhinosinusitis alters facial growth. STUDY DESIGN AND SETTING: This was a retrospective age-matched cohort outcome study performed at a tertiary care hospital. RESULTS: Sixty-seven children participated. There were 46 boys and 21 girls, and the mean age was 3.1 years at presentation and 13.2 years at follow-up. There were 46 children who underwent FES surgery and 21 children who did not undergo FES surgery. Quantitative anthropomorphic analysis was performed using 12 standard facial measurements. A facial plastic expert performed qualitative facial analysis. Both quantitative and qualitative analyses showed no statistical significance in facial growth between children who underwent FES surgery and those who did not undergo FES surgery. CONCLUSIONS: In this study, there was no evidence that FES surgery affected facial growth. SIGNIFICANCE: These results will aid physicians when discussing with parents the risks of FES surgery. PMID- 12087330 TI - Minimally invasive approaches to the sphenoid sinus. AB - OBJECTIVE: We sought to develop an algorithm for surgical approaches to the sphenoid sinus. STUDY DESIGN AND SETTING: Retrospective review was conducted of all patients who underwent a sphenoid sinusotomy by the senior author between July 1994 and August 2001. RESULTS: The study population consisted of 141 patients, in whom 5 different surgical approaches were used: transseptal (47 [33.3%]), transnasal (19 [13.5%]), transethmoid (72 [51.1%]), transmaxillary (2 [1.42%]), and external (1 [0.7%]). Of the 47 transseptal approaches, 43 (91.5%) were for extirpation of a neoplasm. In contrast, 60 of 72 (83.3%) transethmoid procedures were for infectious/inflammatory disorders. An endoscopic approach was used for 7 of 8 (87.5%) skull base repairs. Four transsphenoid optic nerve decompressions were performed. The minor and major complication rates were 2.1% and 0.71%, respectively. CONCLUSIONS: The anatomic location of the pathologic process can guide the surgeon in selecting the most appropriate technique. SIGNIFICANCE: Surgical treatment of sphenoid pathology can be safely and successfully performed through a variety of approaches. PMID- 12087331 TI - Office-based treatment of laryngeal papillomatosis with percutaneous injection of cidofovir. AB - OBJECTIVE: Our aim was to report our experience with office-based treatment of severe laryngeal papillomatosis with percutaneous injection of cidofovir in a case series of 5 patients. STUDY DESIGN AND SETTING: We conducted a retrospective review of a case series in a tertiary academic care voice disorders clinic. Adult patients with papillomas of the vocal cords and anterior commissure received percutaneous injection using a point-touch technique. Clinical improvement or remission of the papillomatosis was noted. RESULTS: Before initiation of office treatments, patients required direct laryngoscopy and CO(2) laser ablation of papillomas on average every 2.8 months. There were no complications related to the injection technique. During a treatment period of 7 to 16 months (mean 12 months), a significant reduction in the volume of papillomatosis was achieved in all patients. One patient received 2 treatments and another received 1 treatment in the operating room for final clearance of papillomas. CONCLUSION: Office-based treatment of adult patients with anterior laryngeal papillomatosis using percutaneous injection of cidofovir reduces the need for repeated direct laryngoscopy and laser ablation under general anesthesia. SIGNIFICANCE: Percutaneous injection treatment with cidofovir is a useful adjunct to direct laryngoscopy and laser ablation in the treatment of laryngeal papillomatosis. PMID- 12087332 TI - The association between laryngeal pseudosulcus and laryngopharyngeal reflux. AB - BACKGROUND: A pattern of edema on the ventral surface of the vocal fold, called pseudosulcus vocalis, was described in 1995. It refers to infraglottic edema extending from the anterior commissure to the posterior larynx. It can be differentiated from sulcus vergeture, which is caused by adhesion of the vocal fold epithelium to the vocal ligament. Although it has been related to laryngopharyngeal reflux (LPR), this has not been thoroughly investigated. PURPOSE: Our goal was to evaluate the association between pseudosulcus and LPR. METHODS: Thirty patients with a clinical diagnosis of LPR confirmed by double probe pH monitoring and 30 controls without LPR were enrolled. The prevalence of pseudosulcus was determined with fiberoptic laryngoscopy. RESULTS: Seventy percent (21 of 30) of patients with lpr and 30% (9 of 30) of controls had pseudosulcus. Patients with pseudosulcus were 2.3 times more likely to have pH documented LPR (95% confidence interval 1.3-4.2). The sensitivity and specificity of pseudosulcus in the diagnosis of LPR are 70% and 77%, respectively. CONCLUSIONS: Pseudosulcus is highly correlated with pH-documented LPR (P < 0.001). The presence of pseudosulcus is suggestive of LPR. PMID- 12087333 TI - Percutaneous laryngeal collagen augmentation for treatment of parkinsonian hypophonia. AB - OBJECTIVE: Our goal was to determine whether percutaneous laryngeal collagen augmentation improves hypophonia in parkinsonian patients. STUDY DESIGN AND SETTING: A retrospective review of 18 patients was performed. Patients and/or caretakers were called on the telephone and asked about their response to the procedure, if any. Videostroboscopic examinations for all patients were reviewed. RESULTS: Of 18 patients, 11 (61%) reported improvement in their hypophonia for a period of at least 2 months. Five of 7 patients without improvement were relatively aphonic both before and after the procedure. Five of 7 patients without improvement had severe dysphagia, which in 3 necessitated gastrostomy tube placement. Four of 7 patients without improvement were not ambulatory at the time of the procedure. CONCLUSIONS: Percutaneous laryngeal collagen augmentation is an effective treatment for parkinsonian hypophonia in a majority of patients. Patients with advanced neurologic disease with aphonia, difficulty with speech initiation, dysphagia, or ambulatory difficulty are less likely to respond to this procedure and should be so informed. PMID- 12087334 TI - Oncocytic neoplasms of the parotid gland: a 16-year institutional review. AB - BACKGROUND: Oncocytic neoplasms of the parotid gland are a rare collection of salivary gland tumors that include oncocytosis, oncocytoma, and oncocytic carcinoma. Mounting evidence has linked the presence of oncocytes to acquired mitochondrial dysfunction. OBJECTIVES: The study goals were to further delineate this poorly understood group of salivary gland tumors and to search for genetic alterations indicative of mitochondrial dysfunction. METHODS: Cases were identified by search of the surgical pathology archival files from 1984 through 2000. Corresponding medical records were reviewed. Tumor mitochondrial DNA (mtDNA) was evaluated for mtDNA mutations within the control region (C-tract). RESULTS: Twenty-one parotid oncocytic neoplasms were identified. Oncocytoma was the most frequent morphology (62%), followed by oncocytosis (28.5%) and oncocytic carcinoma (9.5%). One specimen displayed synchronous oncocytic morphologies (oncocytoma, oncocytosis, and oncocytic metaplasia). One oncocytoma specimen displayed the mtDNA C-tract alteration. CONCLUSIONS: Oncocytic neoplasia of the parotid gland is a rare form of salivary gland disease with obscure etiology. The presence of multiple oncocytic morphologies in a single specimen is suggestive of transition between forms. Although oncocytic tumorigenesis secondary to acquired mitochondrial dysfunction is a plausible mechanism, few of these tumors actually harbor mtDNA alterations within the control region. PMID- 12087335 TI - Ventilation tube removal: does treatment affect perforation closure? AB - OBJECTIVE: The study goal was to determine whether treatment of the tympanic membrane at the time of ventilation tube (VT) removal affects closure of the perforation. STUDY DESIGN: We conducted a retrospective chart review and telephone follow-up of children who underwent VT removal from 1995 through 1998. RESULTS: Among 109 patients (162 ears; 58% male), the mean age at VT removal was 6.7 years. Most VTs (59%) were T-tubes, and most (91%) were removed because of prolonged retention (mean 2.3 years). After VT removal, 111 ears (69%) received treatment, most commonly (44%) with 25% trichloroacetic acid (TCA). At the latest follow-up, 151 of the ears (93%) had healed without additional treatment. Treatment failure occurred more frequently in ears not initially treated with TCA (TCA 3% failure, other treatment 13%, no treatment 8%) and in ears with VTs removed because of otorrhea. CONCLUSIONS: Most perforations healed. TCA may promote closure, but a large randomized clinical trial is needed to eliminate physician bias. PMID- 12087336 TI - Skull base cerebrospinal fluid fistulas: a comprehensive diagnostic algorithm. AB - OBJECTIVE: Our goal was to assess the efficacy of current diagnostic modalities in the management of skull base cerebrospinal fluid (CSF) fistulas. METHODS: We conducted a retrospective review of all patients presenting to our institution over the past 6 years with skull base CSF fistulas. RESULTS: Fifty-two patients were included in the study. beta2-Transferrin analysis of collected specimen was the most efficacious means of confirming a CSF leak. High-resolution computed tomography was the most informative radiographic study, yielding a sensitivity and an accuracy of 87%. Magnetic resonance cisternography, yielding a sensitivity and an accuracy of 78%, was instrumental in localizing the site of leak for a few cases but was most commonly corroborative. Using a graduated diagnostic approach, successful repair was attained in 88% of cases after 1 attempt and 98% after 1 or 2 attempts. CONCLUSION: For patients with skull base CSF fistulas, a graduated diagnostic approach with emphasis on confirmation of leak by beta2-transferrin analysis and precise localization by high-resolution computed tomography is both efficacious and cost effective. PMID- 12087337 TI - Prevalence of heat shock protein in patients with Meniere's disease and allergy. AB - OBJECTIVE: The study purpose was to investigate the prevalence of elevated heat shock protein 70 (HSP-70) in patients with Meniere's disease who have milk allergy compared with those who are not allergic to milk. METHODS: Fifty-five patients with Meniere's disease and allergy in whom milk allergy had been confirmed by intradermal progressive dilutional food testing or skin testing to milk antigen were included. Blood serum was tested for HSP-70 elevation with a Western blot assay using bovine renal extract. The 29 women and 26 men ranged in age from 29 to 76 years (mean age 52.8 years). Forty percent of the patients had bilateral Meniere's disease. RESULTS: Overall prevalence of HSP-70 elevation was 29.1%. This was higher in bilateral patients (50%) than unilateral patients (15%) (P 60 dB, with extensive outer hair cell loss throughout the organ of Corti but particularly in the basal turn. CONCLUSION: These results demonstrate that topical D-methionine provides excellent otoprotection against cisplatin-induced ototoxicity both electrophysiologically and structurally. PMID- 12087339 TI - Embryonal rhabdomyosarcoma of the middle ear. PMID- 12087340 TI - Schwannoma (neurilemmoma) of the palatine tonsil. PMID- 12087341 TI - Pleomorphic adenoma of the buccal space. PMID- 12087342 TI - Safety of radiofrequency volumetric tissue reduction in sleep-disordered breathing. PMID- 12087343 TI - Are herbal products dietary supplements or drugs? An important question for public safety. PMID- 12087344 TI - Effect of St John's wort on the pharmacokinetics of fexofenadine. AB - BACKGROUND: St John's wort is a popular over-the-counter dietary supplement and herbal remedy that has been implicated in drug interactions with several substrates of P-glycoprotein. The effect of St John's wort on P-glycoprotein activity in vivo was examined with use of fexofenadine as selective probe drug. METHODS: A 3-period, open-label, fixed-schedule study design was used. Fexofenadine, 60 mg, was administered orally before administration of St John's wort, with a single dose of St John's wort (900 mg), and after 2 weeks of treatment with St John's wort (300 mg 3 times a day) to determine P-glycoprotein activity. RESULTS: A single dose of St John's wort significantly (P <.05) increased the maximum plasma concentration of fexofenadine by 45% and significantly (P <.05) decreased the oral clearance by 20%, with no change in half-life or renal clearance. Long-term administration of St John's wort did not cause a significant change in fexofenadine disposition relative to the untreated phase. Compared with the single-dose treatment phase, long-term St John's wort caused a significant 35% decrease (P <.05) in maximum plasma concentration and a significant 47% increase (P <.05) in fexofenadine oral clearance. CONCLUSIONS: A single dose of St John's wort resulted in a significant inhibition of intestinal P-glycoprotein. Long-term treatment with St John's wort reversed the changes in fexofenadine disposition observed with single-dose administration. PMID- 12087345 TI - Pharmacology of ephedra alkaloids and caffeine after single-dose dietary supplement use. AB - OBJECTIVE: Serious cardiovascular toxicity has been reported in people taking dietary supplements that contain ma huang (Ephedra) and guarana (caffeine). We assessed the pharmacokinetics and pharmacodynamics of a dietary supplement that contains these herbal stimulants. METHODS: Eight healthy adults received a single oral dose of a thermogenic dietary supplement labeled to contain 20 mg ephedrine alkaloids and 200 mg caffeine after an overnight fast. Serial plasma and urine samples were analyzed by use of liquid chromatography-tandem mass spectrometry for ephedrine alkaloid and caffeine concentrations, and heart rate and blood pressure were monitored for 14 hours. RESULTS: Plasma clearance and elimination half-lives for ephedrine, pseudoephedrine, and caffeine were comparable to published values reported for drug formulations. A prolonged half-life of ephedrine and pseudoephedrine was observed in 1 subject with the highest urine pH. Mean systolic blood pressure increased significantly to a maximum of 14 mm Hg above baseline at 90 minutes after ingestion (P <.001). There was a lag in the mean heart rate response that reached a maximum change of 15 beats/min above baseline at 6 hours after ingestion (P <.001). Diastolic blood pressure changes were insignificant. Two subjects who were taking oral contraceptives had longer caffeine half-lives (15.5 +/- 0.3 hours versus 5.6 +/- 1.7 hours) and lower values for oral clearance (0.34 +/- 0.01 mL/min. kg versus 0.99 +/- 0.41 mL/min. kg) than subjects who were not taking oral contraceptives. CONCLUSIONS: Botanical stimulants have disposition characteristics similar to their pharmaceutical counterparts, and they can produce significant cardiovascular responses after a single dose. PMID- 12087346 TI - Bivalirudin pharmacokinetics and pharmacodynamics: effect of renal function, dose, and gender. AB - BACKGROUND: These studies were conducted to determine whether bivalirudin clearance and pharmacodynamics are dependent on dose, renal function, or gender. METHODS: Two studies were performed. The first comprised 25 patients who were undergoing percutaneous coronary intervention-8 with normal renal function, 11 with mild renal impairment, and 6 with moderate renal impairment. Each patient received a bolus dose of bivalirudin (1 mg/kg) followed by an infusion (2.5 mg/kg per hour for 4 of 6 hours, then 0.5 mg/kg per hour for 4 of 6 hours). The second study enrolled 8 volunteers with severe renal impairment who received a bivalirudin bolus of 1 mg/kg, followed by an infusion of 0.5 mg/kg per hour for 10 hours. Bivalirudin in plasma and urine was assayed with a newly developed, highly specific liquid chromatography-mass spectrometry assay. RESULTS: Clearances at the two infusion doses did not differ significantly (3.23 mL/min per kilogram and 3.16 mL/min per kilogram). There was no statistically significant difference in area under the concentration-time curve (AUC) and in plasma clearance between patients with normal renal function and those with mild renal impairment. Patients with moderate and severe renal impairment had reductions in plasma clearance of 21% and 24%, respectively. The level of anticoagulation(activated clotting time) was similar between groups. There was no difference between male and female patients. CONCLUSION: The clearance of bivalirudin is dependent on renal function but independent of dose and gender. Approximately 20% of unchanged drug is cleared via the kidney, and the remainder presumably undergoes proteolysis intracellularly. The pharmacodynamics of bivalirudin are dose-dependent and gender-independent. Bivalirudin kinetics are linear in the dose ranges that are used in percutaneous coronary intervention and that are under investigation for use in acute coronary syndromes. PMID- 12087347 TI - Angiotensin II attenuates the vasodilating effect of a nitric oxide donor, glyceryl trinitrate: roles of superoxide and angiotensin II type 1 receptors. AB - OBJECTIVE: The development of tolerance to organic nitrates limits their usefulness in the treatment of heart disease. Activation of the renin-angiotensin system by heart failure itself and by nitrate therapy may be one possible mechanism underlying nitrate tolerance. We investigated the effect of subpressor doses of angiotensin II on the vasodilating effect of glyceryl trinitrate in human forearm resistance vessels of healthy male subjects by using venous occlusion strain-gauge plethysmography. METHODS: Glyceryl trinitrate was infused intra-arterially with angiotensin II or vehicle. The effect of blockade of angiotensin II type 1 receptors by candesartan or an antioxidant, vitamin C, on the interaction between angiotensin II and glyceryl trinitrate was also investigated. RESULTS: Angiotensin II infused at 5 pmol/min significantly attenuated the vasodilating effect of glyceryl trinitrate (mean +/- standard deviation [SD] of percentage change in forearm blood flow [FBF]: 28% +/- 20%, 79% +/- 59%, and 208% +/- 72% at 100, 250, and 1000 ng/min of glyceryl trinitrate with placebo; 8% +/- 18%, 47% +/- 41%, and 173% +/- 98% with angiotensin II at 1 pmol/min; and 2% +/- 27%, 39% +/- 40%, and 132% +/- 74% with angiotension II at 5 pmo;/min; P =.0259). Either a single dose of candesartan or coinfusion with vitamin C abolished the angiotensin II-induced attenuation of vasodilation of glyceryl trinitrate. CONCLUSION: Our results suggest that angiotensin II may attenuate the arterial vasodilating effect of glyceryl trinitrate through angiotensin type 1 receptors and presumably through receptor-mediated superoxide production, which may be relevant to the development of nitrate tolerance. PMID- 12087348 TI - Diadenosine pentaphosphate vasodilates the forearm vascular bed: inhibition by theophylline and augmentation by dipyridamole. AB - BACKGROUND: In rats, diadenosine pentaphosphate (AP(5)A) has been implicated in the pathogenesis of essential hypertension. This study describes for the first time the vasomotor action of AP(5)A in humans by means of the "perfused forearm technique." RESULTS: AP(5)A evoked a dose-dependent forearm vasodilator response equal to that of adenosine but less than that of adenosine triphosphate (ATP) at equimolar doses. The P(1)-purinoceptor antagonist theophylline (0.28 micromol/min per deciliter) reduced the percentage decrease in forearm vascular resistance (FVR) to AP(5)A (0.6, 6, and 20 nmol/min/dL): -8% +/- 6%, -50% +/- 6%, and -68% +/- 4% during saline solution versus -7% +/- 4%, -33% +/- 5%, and -45% +/- 6% during theophylline (mean +/- standard error [SE]; ANOVA for repeated measures; P <.05 for the interaction between purine dose and theophylline; n = 10). An inhibitor of equilibrative nucleoside transport, dipyridamole (7.4 nmol/min per deciliter), augmented the AP(5)A (0.6 and 6 nmol/min per deciliter)-induced reduction in FVR as follows: -34% +/- 6% and -67% +/- 5% during saline versus 49% +/- 5% and -80% +/- 3% during dipyridamole (P <.05 for the effect of dipyridamole; n = 6). The bivalent cation chelator ethylenediaminetetra-acetic acid (EDTA) inhibited the rapid degradation of AP(5)A in vitro. In vivo, the highest tolerated intra-arterial EDTA dose was not sufficient to inhibit AP(5)A metabolism. CONCLUSION: Intra-arterial AP(5)A caused a dose-dependent reduction in FVR. This is, at least in part, mediated by its degradation product adenosine. The data do not support an in vivo vasoconstrictor action of AP(5)A, and as such AP(5)A does not seem likely to contribute to the pathogenesis of primary hypertension in humans. PMID- 12087349 TI - Lessons learned from a phase III population pharmacokinetic study of cariporide in coronary artery bypass graft surgery. AB - BACKGROUND: Cariporide (HOE642) is a recently developed inhibitor of the myocardial sodium-hydrogen exchange system. The clinical effects of sodium hydrogen exchange inhibition in patients at high risk for myocardial cell necrosis were investigated in the GUARDIAN trial (n = 11,590 patients). Although the trial did not show a significant benefit of cariporide over placebo in the overall population, a 25% relative risk reduction in the primary end point of death or myocardial infarction (12.1% for the highest tested cariporide dose of 120 mg 3 times a day versus 16.2% for placebo; P =.03) was observed in the subpopulation of patients who underwent bypass surgery. OBJECTIVE: Our objective was to identify an optimal dosing regimen that might offer increased protection during the period of highest risk. METHODS: A population pharmacokinetic model of cariporide was developed with use of data from phase I studies. After adequate predictability was shown for the patients in the pharmacokinetic substudy of the GUARDIAN trial (n = 269 patients), the model was used to predict the individual pharmacokinetic profile in the remaining patients. These predicted concentrations were used to calculate the mean concentration during the period of coronary artery bypass graft surgery (the acute risk period in patients who receive coronary artery bypass grafts), and this mean concentration was used as predictor variable in a time-to-event analysis. RESULTS: A mixture of two Weibull functions adequately described the time course of the observed sum of the acute and chronic hazard rate. The calculated mean concentration during the period of surgery was an adequate predictor for the probability of an event in the acute risk period. The estimated minimal effective mean concentration was 0.5 mg/L. CONCLUSIONS: A dosing regimen with a loading dose of an infusion of 120 mg/h for 1 hour followed by an infusion of 20 mg/h for 47 hours should achieve stable exposure above the estimated minimal effective concentration in more than 95% of patients during and after coronary artery bypass graft surgery. PMID- 12087350 TI - Renal and vascular effects of S21402, a dual inhibitor of angiotensin-converting enzyme and neutral endopeptidase, in healthy subjects with hypovolemia. AB - OBJECTIVE: To examine the mechanism of action of dual inhibitors of angiotensin converting enzyme (ACE) and neutral endopeptidase, also called vasopeptidase inhibitors, we compared the effects of S21402 [(2S)-2-[(2S,3R)-2-thiomethyl-3 phenylbutanamido]propionic acid], which belongs to this pharmacologic class, with those of captopril, an ACE inhibitor, on blood pressure, endocrine parameters, and renal in healthy subjects with hypovolemia. METHODS: Ten subjects participated to this double-blind, 2-period, randomized, crossover study. Hypovolemia was induced in these subjects with a 7-day treatment of hydrochlorothiazide. They received a single oral dose of 50 mg captopril or 250 mg S21402 on the last day of diuretic treatment. Blood pressure was measured, and urine and blood samples were collected before and during a 12-hour period after drug administration. RESULTS: The plasma angiotensin II/angiotensin I ratio and aldosterone concentration decreased to the same degree with both drugs, 3 hours after dosing. Compared with captopril, S21402 increased levels of plasma atrial natriuretic peptide (P <.05) and urinary cyclic guanosine monophosphate (P <.001); these increases were the result of inhibition of neutral endopeptidase activity (P <.001). The increase in plasma renin concentration related to ACE inhibition was less marked (P <.001) after S21402 than after captopril. S21402, but not captopril, increased urinary sodium excretion (P <.05), without modifying blood pressure and creatinine clearance, whereas blood pressure transiently fell after captopril administration (P <.05). CONCLUSIONS: In healthy hypovolemic subjects, the vasopeptidase inhibitor S21402 exhibits a natriuretic effect and does not affect blood pressure or glomerular filtration rate. In these conditions, the acute endocrine, vascular, and renal effects of vasopeptidase inhibition differ from those of ACE inhibition. PMID- 12087351 TI - Genetic polymorphisms of glutathione S-transferase A1, the major glutathione S transferase in human liver: consequences for enzyme expression and busulfan conjugation. AB - BACKGROUND: High-dose busulfan is widely used as part of conditioning regimens for patients who are undergoing hematopoietic stem cell or bone marrow transplantation. High plasma concentrations of busulfan have been linked to the occurrence of hepatic venoocclusive disease (VOD), a severe complication associated with a high mortality. Because conjugation with glutathione, the major route of biotransformation of busulfan, is predominantly catalyzed by the isozyme glutathione S-transferase A1 (GSTA1), we hypothesized that low expression or function of GSTA1 in liver caused by genetic polymorphisms may be the mechanism underlying VOD. METHODS: Immunoblot analysis of GSTA and measurement of busulfan glutathione conjugation by liquid chromatography-mass spectrometry were performed in 48 normal human liver samples. To search for polymorphisms, the complete GSTA1 coding regions and the promoter fragment were sequenced. All results were compared by multivariate analysis. RESULTS: Absolute levels of GSTA protein and formation rates of busulfan-glutathione conjugate displayed a 7- and 8-fold range, from 240 to 1600 pmol/mg and 25 to 205 pmol/min per milligram of total cytosolic protein, respectively, and correlate (r2 = 0.49, P <.0001). A total of 8 single nucleotide polymorphisms (SNPs) of GSTA1 were identified, 1 of which was a silent mutation in exon 5 (A375G); all others were found in the promoter region. Haplotype analysis revealed the existence of 5 defined alleles. There was no significant relationship between any of the GSTA1 SNPs or haplotypes and either hepatic glutathione S-transferase A (GSTA) expression or GSTA1 function. CONCLUSIONS: The identified GSTA1 polymorphisms are not likely to be related to the VOD because they do not appear to be associated with changes in GSTA expression or function. Compared with other members of the GST family, GSTA1 displays surprisingly little variation. PMID- 12087352 TI - Effect of interferon alpha-ribavirin bitherapy on cytochrome P450 1A2 and 2D6 and N-acetyltransferase-2 activities in patients with chronic active hepatitis C. AB - BACKGROUND: Interferon alpha (IFN-alpha) is thought to be responsible for cytochrome P450 (CYP)-dependent drug interactions mediated by a decrease in CYP activities. OBJECTIVES: The objectives are to determine whether IFN-alpha and ribavirin can alter pretreatment CYP1A2, CYP2D6, CYP3A4 and N-acetyltransferase-2 activities after 1 month of treatment. METHODS: Enzymatic activities were determined among 14 patients with chronic active hepatitis C before IFN-alpha (3. 10(6) U, 3 times a week) and ribavirin introduction and after 1 month of treatment. During both study periods, subjects received 80 mg dextromethorphan and 140 mg caffeine (1,3,7-trimethylxanthine [137X]). CYP3A4, CYP2D6, and NAT2 activities were assessed by use of urinary metabolic ratios of 3 methoxymorphinan/dextromethorphan, dextrorphan/dextromethorphan, and 5 acetylamino-6-formylamino-3-methyluracil (AFMU)/1-methylxanthine(1X). The plasma paraxanthine/caffeine ratio was used to measure CYP1A2 activity. RESULTS: CYP3A4 and CYP2D6 activities tended to increase after 1 month of antiviral therapy, but the change did not reach statistical significance. CYP1A2 and NAT2 activities were not significantly modified after 1 month of antiviral treatment. Pretreatment activities were significantly lower than those previously observed in healthy volunteers for CYP2D6 (mean +/- SD, 148 +/- 139 versus 759 +/- 692; P =.0008) and CYP3A4 (0.18 +/- 0.06 versus 0.52 +/- 0.72; P =.0006). This difference was no longer statistically significant after 1 month of treatment, because CYP2D6 and CYP3A4 activities improved in 7 patients. CONCLUSION: In patients with chronic hepatitis C, pretreatment CYP3A4 and CYP2D6 activities were significantly lower than those observed in healthy volunteers. These differences disappeared after 1 month of antiviral treatment because of the restoration of these CYP activities in about half of the patients. PMID- 12087353 TI - Overanticoagulation associated with combined use of antifungal agents and coumarin anticoagulants. AB - BACKGROUND: Several case reports have associated combined use of coumarins and antifungal agents with overanticoagulation. However, we are not aware of epidemiologic studies that quantify the risk of overanticoagulation caused by antifungal agents. We conducted a follow-up study in a large population-based cohort to investigate the antifungal agents that are associated with overanticoagulation during therapy with coumarins. METHODS: All patients in the Rotterdam Study who were treated with acenocoumarol or phenprocoumon in the study period from April 1, 1991, through Dec 31, 1998, and for whom international normalized ratio data were available were followed up until an INR > or =6.0, the end of their treatment, death, or the end of the study period. Proportional hazards regression analysis was used to estimate the risk of an INR > or =6.0 in relation to concomitant use of an oral anticoagulant and antifungal agents, after adjustment for several potentially confounding factors such as age, gender, hepatic dysfunction, malignancies, and heart failure. RESULTS: Of the 1124 patients in the cohort, 351 had an INR > or =6.0. The incidence rate was 6.9 per 10,000 treatment days. Oral miconazole most strongly increased the risk of overanticoagulation with an adjusted relative risk of 36.6 (95% confidence interval [CI], 12.4-108.0). CONCLUSIONS: In this study among outpatients of an anticoagulant clinic who were on a regimen of coumarins, oral miconazole was especially strongly associated with overanticoagulation. Awareness of these drug interactions and more frequent monitoring of INR values during the initial stages of treatment with some antifungal drugs in patients taking coumarins may minimize the risk of bleeding complications. The concurrent use of miconazole and coumarins should be discouraged. PMID- 12087354 TI - Angiotensin-converting enzyme inhibitor-induced syndrome of inappropriate secretion of antidiuretic hormone: case report and review of the literature. AB - Seventeen cases of severe hyponatremia induced by angiotensin-converting enzyme (ACE) inhibitor therapy have been reported in the literature. The mechanism of severe hyponatremia induced by ACE inhibitor is not clear. A 60-year-old white man with a history of idiopathic dilated cardiomyopathy was treated with enalapril, 20 mg daily, that had been started 2 weeks before heart transplantation. The serum sodium level was 138 mmol/L before initiation of enalapril therapy and 127 mmol/L just before cardiac surgery. In the post-heart transplantation period, enalapril therapy was withdrawn for the perianesthesia period, and the serum sodium level increased from 127 to 140 mmol/L. One month later, viral myocarditis developed in the patient and enalapril was reintroduced at 20 mg daily. Two weeks later, natremia decreased. Enalapril was discontinued. Three days later the serum sodium level rose to 140 mmol/L. Severe symptomatic hyponatremia induced by the syndrome of inappropriate secretion of antidiuretic hormone should be considered a rare but possible complication associated with ACE inhibitor therapy. PMID- 12087355 TI - Promoting language and literacy through reading aloud: the role of the pediatrician. PMID- 12087357 TI - Short interfering RNA confers intracellular antiviral immunity in human cells. AB - Gene silencing mediated by double-stranded RNA (dsRNA) is a sequence-specific, highly conserved mechanism in eukaryotes. In plants, it serves as an antiviral defence mechanism. Animal cells also possess this machinery but its specific function is unclear. Here we demonstrate that dsRNA can effectively protect human cells against infection by a rapidly replicating and highly cytolytic RNA virus. Pre-treatment of human and mouse cells with double-stranded, short interfering RNAs (siRNAs) to the poliovirus genome markedly reduces the titre of virus progeny and promotes clearance of the virus from most of the infected cells. The antiviral effect is sequence-specific and is not attributable to either classical antisense mechanisms or to interferon and the interferon response effectors protein kinase R (PKR) and RNaseL. Protection is the result of direct targeting of the viral genome by siRNA, as sequence analysis of escape virus (resistant to siRNAs) reveals one nucleotide substitution in the middle of the targeted sequence. Thus, siRNAs elicit specific intracellular antiviral resistance that may provide a therapeutic strategy against human viruses. PMID- 12087360 TI - Silicon dreams in the biology lab. PMID- 12087358 TI - Modulation of HIV-1 replication by RNA interference. AB - RNA interference (RNAi) is the process by which double-stranded RNA (dsRNA) directs sequence-specific degradation of messenger RNA in animal and plant cells. In mammalian cells, RNAi can be triggered by 21-nucleotide duplexes of small interfering RNA (siRNA). Here we describe inhibition of early and late steps of HIV-1 replication in human cell lines and primary lymphocytes by siRNAs targeted to various regions of the HIV-1 genome. We demonstrate that synthetic siRNA duplexes or plasmid-derived siRNAs inhibit HIV-1 infection by specifically degrading genomic HIV-1 RNA, thereby preventing formation of viral complementary DNA intermediates. These results demonstrate the utility of RNAi for modulating the HIV replication cycle and provide evidence that genomic HIV-1 RNA, as it exists within a nucleoprotein reverse-transcription complex, is amenable to siRNA mediated degradation. PMID- 12087361 TI - Cell biology gets plugged in. PMID- 12087362 TI - Computerized role models. PMID- 12087363 TI - Anyone for neutrons? PMID- 12087364 TI - Bush's advisers lean towards ban on research cloning. PMID- 12087365 TI - Code of conduct for bioethics branded 'soft' on corporate ties. PMID- 12087366 TI - Scientists jailed for alleged theft from Harvard laboratory. PMID- 12087369 TI - CERN puts research on hold to build collider. PMID- 12087368 TI - Fight against terror 'needs science institute'. PMID- 12087372 TI - NASA launch heralds fresh wave of comet exploration. PMID- 12087374 TI - Altered states. PMID- 12087375 TI - The society of proteins. PMID- 12087376 TI - Conflicts around a study of Mexican crops. PMID- 12087377 TI - Conflicts around a study of Mexican crops. PMID- 12087380 TI - Impact-factor rewards affect Spanish research. PMID- 12087386 TI - Through a lens brightly. PMID- 12087385 TI - Maxwell's demon: slamming the door. PMID- 12087387 TI - Lucky draw in the gene raffle. PMID- 12087388 TI - Thin-film cliffhanger. PMID- 12087390 TI - Plant-microbe interactions: a receptor in symbiotic dialogue. PMID- 12087391 TI - How old roots lose their bounce. PMID- 12087392 TI - Genomic imprinting: piece of cake. PMID- 12087394 TI - Caffeine as a repellent for slugs and snails. PMID- 12087395 TI - Early-pregnancy origins of low birth weight. PMID- 12087396 TI - Sub-laser-cycle electron pulses for probing molecular dynamics. AB - Experience shows that the ability to make measurements in any new time regime opens new areas of science. Currently, experimental probes for the attosecond time regime (10(-18) 10(-15) s) are being established. The leading approach is the generation of attosecond optical pulses by ionizing atoms with intense laser pulses. This nonlinear process leads to the production of high harmonics during collisions between electrons and the ionized atoms. The underlying mechanism implies control of energetic electrons with attosecond precision. We propose that the electrons themselves can be exploited for ultrafast measurements. We use a 'molecular clock', based on a vibrational wave packet in H(2)(+) to show that distinct bunches of electrons appear during electron ion collisions with high current densities, and durations of about 1 femtosecond (10(-15) s). Furthermore, we use the molecular clock to study the dynamics of non-sequential double ionization. PMID- 12087397 TI - Magnification of light from many distant quasars by gravitational lenses. AB - Exceptionally bright quasars with redshifts up to z = 6.28 have recently been discovered. Quasars are thought to be powered by the accretion of gas onto supermassive black holes at the centres of galaxies. Their maximum (Eddington) luminosity depends on the mass of the black hole, and the brighter quasars are inferred to have black holes with masses of more than a few billion solar masses. The existence of such massive black holes poses a challenge to models for the formation of structures in the early Universe, as it requires their formation within one billion years of the Big Bang. Here we show that up to one-third of known quasars with z approximately equal to 6 will have had their observed flux magnified by a factor of ten or more, as a consequence of gravitational lensing by galaxies along the line of sight. The inferred abundance of quasar host galaxies, as well as the luminosity density provided by the quasars, has therefore been substantially overestimated. PMID- 12087398 TI - The nature and transport mechanism of hydrated hydroxide ions in aqueous solution. AB - Compared to other ions, protons (H(+)) and hydroxide ions (OH(-)) exhibit anomalously high mobilities in aqueous solutions. On a qualitative level, this behaviour has long been explained by 'structural diffusion' the continuous interconversion between hydration complexes driven by fluctuations in the solvation shell of the hydrated ions. Detailed investigations have led to a clear understanding of the proton transport mechanism at the molecular level. In contrast, hydroxide ion mobility in basic solutions has received far less attention, even though bases and base catalysis play important roles in many organic and biochemical reactions and in the chemical industry. The reason for this may be attributed to the century-old notion that a hydrated OH(-) can be regarded as a water molecule missing a proton, and that the transport mechanism of such a 'proton hole' can be inferred from that of an excess proton by simply reversing hydrogen bond polarities. However, recent studies have identified OH(-) hydration complexes that bear little structural similarity to proton hydration complexes. Here we report the solution structures and transport mechanisms of hydrated hydroxide, which we obtained from first-principles computer simulations that explicitly treat quantum and thermal fluctuations of all nuclei. We find that the transport mechanism, which differs significantly from the proton hole picture, involves an interplay between the previously identified hydration complexes and is strongly influenced by nuclear quantum effects. PMID- 12087399 TI - Nonlinear dynamics of ice-wedge networks and resulting sensitivity to severe cooling events. AB - Patterns of subsurface wedges of ice that form along cooling-induced tension fractures, expressed at the ground surface by ridges or troughs spaced 10 30 m apart, are ubiquitous in polar lowlands. Fossilized ice wedges, which are widespread at lower latitudes, have been used to infer the duration and mean temperature of cold periods within Proterozoic and Quaternary climates, and recent climate trends have been inferred from fracture frequency in active ice wedges. Here we present simulations from a numerical model for the evolution of ice-wedge networks over a range of climate scenarios, based on the interactions between thermal tensile stress, fracture and ice wedges. We find that short-lived periods of severe cooling permanently alter the spacing between ice wedges as well as their fracture frequency. This affects the rate at which the widths of ice wedges increase as well as the network's response to subsequent climate change. We conclude that wedge spacing and width in ice-wedge networks mainly reflect infrequent episodes of rapidly falling ground temperatures rather than mean conditions. PMID- 12087400 TI - Waning buoyancy in the crustal roots of old mountains. AB - When mountains form through the collision of lithospheric plates, uplift of the Earth's surface is accompanied by thickening of the crust, and the buoyancy of these deep crustal roots (relative to the surrounding mantle) is thought to contribute to the support of mountain topography. Once active tectonism ceases, continuing erosion will progressively wear away surface relief. Here I provide new constraints on how crustal roots respond to erosional unloading over very long timescales. In old collisional mountain belts, ratios of surface relief to the thickness of the underlying crustal root are observed to be smaller than in young mountains. On the basis of gravity data, this trend is best explained by a decrease in the buoyancy of the crustal root with greater age since the most recent mountain-building episode which is consistent with metamorphic reactions produced by long-term cooling. An approximate balance between mountain and root mass anomalies suggests that the continental lithosphere remains weak enough to permit exhumation of crustal roots in response to surface erosion for hundreds of millions of years. The amount of such uplift, however, appears to be significantly reduced by progressive loss of root buoyancy. PMID- 12087401 TI - Bird-like fossil footprints from the Late Triassic. AB - The study of fossilized footprints and tracks of dinosaurs and other vertebrates has provided insight into the origin, evolution and extinction of several major groups and their behaviour; it has also been an important complement to their body fossil record. The known history of birds starts in the Late Jurassic epoch (around 150 Myr ago) with the record of Archaeopteryx, whereas the coelurosaurian ancestors of the birds date back to the Early Jurassic. The hind limbs of Late Triassic epoch theropods lack osteological evidence for an avian reversed hallux and also display other functional differences from birds. Previous references to suggested Late Triassic to Early Jurassic bird-like footprints have been reinterpreted as produced by non-avian dinosaurs having a high angle between digits II and IV and in all cases their avian affinities have been challenged. Here we describe well-preserved and abundant footprints with clearly avian characters from a Late Triassic redbed sequence of Argentina, at least 55 Myr before the first known skeletal record of birds. These footprints document the activities, in an environment interpreted as small ponds associated with ephemeral rivers, of an unknown group of Late Triassic theropods having some avian characters. PMID- 12087402 TI - Overlap of internal models in motor cortex for mechanical loads during reaching. AB - A hallmark of the human motor system is its ability to adapt motor patterns for different environmental conditions, such as when a skilled ice-hockey player accurately shoots a puck with or without protective equipment. Each object (stick, shoulder pad, elbow pad) imparts a distinct load upon the limb, and a key problem in motor neuroscience is to understand how the brain controls movement for different mechanical contexts. We addressed this issue by training non-human primates to make reaching movements with and without viscous loads applied to the shoulder and/or elbow joints, and then examined neural representations in primary motor cortex (MI) for each load condition. Even though the shoulder and elbow loads are mechanically independent, we found that some neurons responded to both of these single-joint loads. Furthermore, changes in activity of individual neurons during multi-joint loads could be predicted from their response to subordinate single-joint loads. These findings suggest that neural representations of different mechanical contexts in MI are organized in a highly structured manner that may provide a neural basis for how complex motor behaviour is learned from simpler motor tasks. PMID- 12087403 TI - Placental-specific IGF-II is a major modulator of placental and fetal growth. AB - Imprinted genes in mammals are expressed from only one of the parental chromosomes, and are crucial for placental development and fetal growth. The insulin-like growth factor II gene (Igf2) is paternally expressed in the fetus and placenta. Here we show that deletion from the Igf2 gene of a transcript (P0) specifically expressed in the labyrinthine trophoblast of the placenta leads to reduced growth of the placenta, followed several days later by fetal growth restriction. The fetal to placental weight ratio is thus increased in the absence of the P0 transcript. We show that passive permeability for nutrients of the mutant placenta is decreased, but that secondary active placental amino acid transport is initially upregulated, compensating for the decrease in passive permeability. Later the compensation fails and fetal growth restriction ensues. Our study provides experimental evidence for imprinted gene action in the placenta that directly controls the supply of maternal nutrients to the fetus, and supports the genetic conflict theory of imprinting. We propose that the Igf2 gene, and perhaps other imprinted genes, control both the placental supply of, and the genetic demand for, maternal nutrients to the mammalian fetus. PMID- 12087404 TI - VEGF regulates haematopoietic stem cell survival by an internal autocrine loop mechanism. AB - Vascular endothelial growth factor (VEGF) is a principal regulator of blood vessel formation and haematopoiesis, but the mechanisms by which VEGF differentially regulates these processes have been elusive. Here we describe a regulatory loop by which VEGF controls survival of haematopoietic stem cells (HSCs). We observed a reduction in survival, colony formation and in vivo repopulation rates of HSCs after ablation of the VEGF gene in mice. Intracellularly acting small-molecule inhibitors of VEGF receptor (VEGFR) tyrosine kinase dramatically reduced colony formation of HSCs, thus mimicking deletion of the VEGF gene. However, blocking VEGF by administering a soluble VEGFR-1, which acts extracellularly, induced only minor effects. These findings support the involvement in HSC survival of a VEGF-dependent internal autocrine loop mechanism (that is, the mechanism is resistant to inhibitors that fail to penetrate the intracellular compartment). Not only ligands selective for VEGF and VEGFR-2 but also VEGFR-1 agonists rescued survival and repopulation of VEGF deficient HSCs, revealing a function for VEGFR-1 signalling during haematopoiesis. PMID- 12087405 TI - A plant receptor-like kinase required for both bacterial and fungal symbiosis. AB - Most higher plant species can enter a root symbiosis with arbuscular mycorrhizal fungi, in which plant carbon is traded for fungal phosphate. This is an ancient symbiosis, which has been detected in fossils of early land plants. In contrast, the nitrogen-fixing root nodule symbioses of plants with bacteria evolved more recently, and are phylogenetically restricted to the rosid I clade of plants. Both symbioses rely on partially overlapping genetic programmes. We have identified the molecular basis for this convergence by cloning orthologous SYMRK ('symbiosis receptor-like kinase') genes from Lotus and pea, which are required for both fungal and bacterial recognition. SYMRK is predicted to have a signal peptide, an extracellular domain comprising leucine-rich repeats, a transmembrane and an intracellular protein kinase domain. Lotus SYMRK is required for a symbiotic signal transduction pathway leading from the perception of microbial signal molecules to rapid symbiosis-related gene activation. The perception of symbiotic fungi and bacteria is mediated by at least one common signalling component, which could have been recruited during the evolution of root nodule symbioses from the already existing arbuscular mycorrhiza symbiosis. PMID- 12087406 TI - A receptor kinase gene regulating symbiotic nodule development. AB - Leguminous plants are able to establish a nitrogen-fixing symbiosis with soil bacteria generally known as rhizobia. Metabolites exuded by the plant root activate the production of a rhizobial signal molecule, the Nod factor, which is essential for symbiotic nodule development. This lipo-chitooligosaccharide signal is active at femtomolar concentrations, and its structure is correlated with host specificity of symbiosis, suggesting the involvement of a cognate perception system in the plant host. Here we describe the cloning of a gene from Medicago sativa that is essential for Nod-factor perception in alfalfa, and by genetic analogy, in the related legumes Medicago truncatula and Pisum sativum. The identified 'nodulation receptor kinase', NORK, is predicted to function in the Nod-factor perception/transduction system (the NORK system) that initiates a signal cascade leading to nodulation. The family of 'NORK extracellular-sequence like' (NSL) genes is broadly distributed in the plant kingdom, although their biological function has not been previously ascribed. We suggest that during the evolution of symbiosis an ancestral NSL system was co-opted for transduction of an external ligand, the rhizobial Nod factor, leading to development of the symbiotic root nodule. PMID- 12087409 TI - Surveying the framework. PMID- 12087407 TI - Structure of a bacterial quorum-sensing transcription factor complexed with pheromone and DNA. AB - Many proteobacteria are able to monitor their population densities through the release of pheromones known as N-acylhomoserine lactones. At high population densities, these pheromones elicit diverse responses that include bioluminescence, biofilm formation, production of antimicrobials, DNA exchange, pathogenesis and symbiosis. Many of these regulatory systems require a pheromone dependent transcription factor similar to the LuxR protein of Vibrio fischeri. Here we present the structure of a LuxR-type protein. TraR of Agrobacterium tumefaciens was solved at 1.66 A as a complex with the pheromone N-3-oxooctanoyl L-homoserine lactone (OOHL) and its TraR DNA-binding site. The amino-terminal domain of TraR is an alpha/beta/alpha sandwich that binds OOHL, whereas the carboxy-terminal domain contains a helix turn helix DNA-binding motif. The TraR dimer displays a two-fold symmetry axis in each domain; however, these two axes of symmetry are at an approximately 90 degree angle, resulting in a pronounced overall asymmetry of the complex. The pheromone lies fully embedded within the protein with virtually no solvent contact, and makes numerous hydrophobic contacts with the protein as well as four hydrogen bonds: three direct and one water-mediated. PMID- 12087410 TI - Lack of IgA in C(mu)-deficient patients. PMID- 12087412 TI - RNA interference: antiviral defense and genetic tool. PMID- 12087413 TI - CD8(+) T cell responses: it's all downhill after their prime. PMID- 12087414 TI - Innate defense evicts bacterial squatters. PMID- 12087415 TI - Thymus by numbers. PMID- 12087416 TI - TSLP: finally in the limelight. PMID- 12087417 TI - To bend or not to bend... PMID- 12087418 TI - Interfering HIV. PMID- 12087419 TI - CTLA-4: new insights into its biological function and use in tumor immunotherapy. AB - The discovery of multiple costimulatory cell surface molecules that influence the course of T cell activation has increased our appreciation of the complexity of the T cell response. It remains clear, however, that CD28 and cytotoxic T lymphocyte antigen 4 (CTLA-4) are the critical costimulatory receptors that determine the early outcome of stimulation through the T cell antigen receptor (TCR). Details of how the T cell integrates TCR stimulation with the costimulatory signals of CD28 and the inhibitory signals of CTLA-4 remain to be established, but unique features of the cell biology of CTLA-4 provide important insights into its function. We summarize here recent findings that suggest a previously unrecognized role for CTLA-4 in the regulation of T cell responses. We also describe preclinical and clinical results that indicate manipulation of CTLA 4 has considerable promise as a strategy for the immunotherapy of cancer. PMID- 12087420 TI - Toxicology and carcinogenesis studies of o-nitrotoluene sulfone (CAS no. 88-72-2) in F344/N rats and B6C3F(1) mice (feed studies). AB - [structure: see text] o-Nitrotoluene is used to synthesize agricultural and rubber chemicals, azo and sulfur dyes, and dyes for cotton, wool, silk, leather, and paper. o-Nitrotoluene was nominated for study by NIOSH and the NTP based on its considerable human exposure as well as the absence of long-term studies of carcinogenicity in rodents. Male and female F344/N rats and B6C3F1 mice were exposed to o-nitrotoluene (greater than 99% pure) in feed for 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, cultured Chinese hamster ovary cells, rat and mouse bone marrow cells, and mouse peripheral blood erythrocytes. 2-YEAR STUDY IN RATS: In the core study, groups of 60 male and 60 female rats were fed diets containing 625, 1,250, or 2,000 ppm o-nitrotoluene (equivalent to average daily doses of approximately 25, 50, or 90 mg o nitrotoluene/kg body weight to males and 30, 60, or 100 mg/kg to females) for 105 weeks. In a 3-month stop-exposure study, groups of 70 male rats were fed diets containing 2,000 or 5,000 ppm o-nitrotoluene (equivalent to average daily doses of approximately 125 or 315 mg/kg) for 13 weeks followed by undosed feed for the remainder of the study. A group of 70 male rats receiving undosed feed served as a control group for both male rat studies; 60 female rats receiving undosed feed were the control group for the female core study. Ten control males and 10 males from each stop-exposure group were sacrificed at 3 months. Survival, Body Weights, and Feed Consumption: All 2,000 ppm core study, all 5,000 ppm stop exposure, and all but three core study 1,250 ppm male rats died before the end of the studies. Survival of 625 ppm core study and 2,000 ppm stop-exposure males and of 2,000 ppm females was significantly less than that of the controls. Mean body weights of all exposed groups of males except the 625 ppm group were generally less than those of the controls throughout the study. Mean body weights of 2,000 ppm females were less than those of the controls during year 2 of the study. Feed consumption by exposed groups of rats was similar to that by the controls. Biomarkers of Exposure: Three urinary metabolites were followed during the study as biomarkers of exposure. The ratios of o-nitrobenzoic acid to creatinine and of o-nitrobenzylmercapturic acid to creatinine determined at 2 weeks and at 3, 12, and 18 months were linearly related to exposure concentration in males and females. The ratio of o-aminobenzoic acid to creatinine was not related to exposure concentration. Pathology Findings: The incidences of malignant mesothelioma in male rats occurred with positive trends in both the core and stop exposure studies and were significantly greater in exposed groups than in the controls. Incidences of subcutaneous skin neoplasms (fibroma, fibrosarcoma, and lipoma) were increased in exposed groups of males, while the incidences of fibroma or fibrosarcoma (combined) were increased in exposed females. In all exposed groups of males and females except 2,000 ppm core study males, the incidences of mammary gland fibroadenoma were significantly increased. The incidences of mammary gland hyperplasia were significantly increased in 625 and 1,250 ppm females. Increased incidences of mesothelioma, skin neoplasms, and mammary gland fibroadenoma in the stop-exposure males indicated that 3 months of dosing were sufficient to produce a carcinogenic effect. Liver weights of 5,000 ppm stop-exposure males were significantly greater than those of the controls at 3 months. The incidences of hepatocellular adenoma in 2,000 ppm core study males and females and of hepatocellular adenoma or carcinoma (combined) in 2,000 ppm core study and 5,000 ppm stop-exposure males were significantly increased. Cholangiocarcinoma occurred in three 5,000 ppm stop-exposure males, and a single hepatocholangiocarcinoma occurred in a 625 ppm male and in a 2,000 ppm core study male. Nonneoplastic lesions of the liver included eosinophilic, mixed cell, and clear cell foci in exposed groups of males and females and mixed cell infiltrate in exposed males and basophilic focus in exposed females. The incidences of alveolar/bronchiolar adenoma and alveolar/bronchiolar adenoma or carcinoma (combined) were significantly increased in 5,000 ppm stop-exposure males, as were alveolar/bronchiolar hyperplasia in most exposed groups of males and females. The incidences of hematopoietic cell proliferation of the spleen and of hyperplasia of the mandibular lymph node (females) and bone marrow were increased in exposed groups of males at 3 months and/or 2 years and in exposed groups of females at 2 years. The incidences of mononuclear cell leukemia were significantly decreased in all groups of males exposed to 1,250 ppm or greater and in all exposed groups of females; the incidence of testicular interstitial cell adenoma was significantly decreased in 5,000 ppm stop-exposure males. 2-YEAR STUDY IN MICE: Groups of 60 male and 60 female mice were fed diets containing 0, 1,250, 2,500, or 5,000 ppm o-nitrotoluene (equivalent to average daily doses of approximately 165, 360, or 700 mg/kg to males and 150, 320, or 710 mg/kg to females) for 105 weeks. Survival, Body Weights, and Feed Consumption: All 2,500 and 5,000 ppm males died before the end of the study. Survival of 1,250 ppm males and 5,000 ppm females was significantly less than that of the controls. Mean body weights of exposed males and 5,000 ppm females were generally less than those of the controls throughout the study, and those of 2,500 ppm females were less during the second year of the study. Feed consumption by 5,000 ppm males was less than that by the controls. Biomarkers of Exposure: Three urinary metabolites were followed during the study as biomarkers of exposure. The ratios of o-nitrobenzoic acid to creatinine determined at 2 weeks and at 3, 12, and 18 months were linearly related to exposure concentration in males and females. The concentrations of o-nitrobenzylmercapturic acid and o-aminobenzoic acid were below the limit of quantitation at most time points. Pathology Findings: The incidences of hemangiosarcoma in all exposed groups of males and in 5,000 ppm females were significantly greater than those in the controls. Large intestine (cecum) carcinomas were observed in all exposed groups except 5,000 ppm males. The incidences of hepatocellular neoplasms were significantly increased in 2,500 and 5,000 ppm females. Nonneoplastic liver lesions including eosinophilic and basophilic foci and minimal to mild necrosis were enhanced in exposed males and females. Also present were focal hepatocyte syncytial alteration in exposed males and hepatocyte necrosis and focal hepatocyte cytoplasmic vacuolization in 5,000 ppm females. Renal tubule pigmentation occurred more frequently in exposed groups of males and in 5,000 ppm females than in the controls. Olfactory epithelial degeneration occurred in every male and female mouse exposed to 2,500 or 5,000 ppm, and the severity of this lesion increased with increasing exposure concentration. GENETIC TOXICOLOGY: o-Nitrotoluene was not mutagenic in any of several strains of S. typhimurium, with or without metabolic activation enzymes (S9). Sister chromatid exchanges were significantly increased in cultured Chinese hamster ovary cells following exposure to o-nitrotoluene in the presence of S9; an equivocal response was seen without S9. o-Nitrotoluene did not induce chromosomal aberrations in cultured Chinese hamster ovary cells, with or without S9. o-Nitrotoluene did not induce a significant increase in the frequency of micronuclei in bone marrow polychromatic erythrocytes of male rats or male mice when administered by intraperitoneal injection. Results of a peripheral blood micronucleus test were equivocal for male mice and negative for female mice administered o-nitrotoluene in feed for 13 weeks. CONCLUSIONS: Under the conditions of these studies, there was clear evidence of carcinogenic activity* of o-nitrotoluene in male rats based on increased incidences of malignant mesothelioma, subcutaneous skin neoplasms, mammary gland fibroadenoma, and liver neoplasms. The increased incidences of lung neoplasms in male rats were also considered to be exposure related. There was clear evidence of carcinogenic activity of o-nitrotoluene in female rats based on increased incidences of subcutaneous skin neoplasms and mammary gland fibroadenoma. The increased incidence of hepatocellular adenoma in female rats was also considered to be exposure related. There was clear evidence of carcinogenic activity of -o nitrotoluene in male and female mice based on increased incidences of hemangiosarcoma, carcinoma of the large intestine (cecum), and hepatocellular neoplasms (females only). Exposure to o--nitrotoluene caused increased incidences of nonneoplastic lesions of the mammary gland (females only), liver, bone marrow, spleen, lung, and mandibular lymph node (females only) in male and female rats and of the liver, kidney, and nose in male and female mice. Decreased incidences of mononuclear cell leukemia occurred in exposed groups of rats; the incidence of testicular interstitial cell adenoma was decreased in exposed male rats. [tables: see text] PMID- 12087421 TI - Toxicology and carcinogenesis studies of p,p'-dichlorophenyl sulfone (CAS No. 80 07-9) in F344/N rats and B6C3F1 mice (feed studies). AB - p,pN-Dichlorodiphenyl sulfone is used as a starting material in the production of polysulfones and polyethersulfones and as a component in reactive dyes in the textile industry; it is also a by-product of pesticide production. p,pN Dichlorodiphenyl sulfone was nominated for study by the National Cancer Institute because of its history of high production and use, the prospect of increased production and use, and the absence of adequate toxicity testing. Male and female F344/N rats and B6C3F1 mice were exposed top,pN-dichlorodiphenyl sulfone (greater than 99% pure)in feed for 14 weeks or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium,cultured Chinese hamster ovary cells, and mouse bone marrow. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female F344/N rats were fed diets containing 0, 30, 100, 300, 1,000, or 3,000 ppm p,pN dichlorodiphenyl sulfone (equivalent to average daily doses of approximately 2, 6, 19, 65, or 200 mgp,pN-dichlorodiphenyl sulfone/kg body weight) for 14 weeks. All rats survived until the end of the study. Mean body weights of groups exposed to 300 ppm or greater were significantly less than those of the controls. Liver weights of groups exposed to 100 ppm or greater and kidney weights of 1,000 and 3,000 ppm male rats were significantly greater than those of the controls. Centrilobular hepatocyte hypertrophy of the liver was observed in most male rats exposed to 100 ppm or greater and in all female rats exposed to 300 ppm or greater, and the severities were increased in 300 ppm males and 1,000 and 3,000 ppm males and females. The incidences of nephropathy in 1,000 and 3,000 ppm female rats were significantly increased. Dose-related increases in severity of nephropathy were observed in male rats. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female B6C3F1 mice were fed diets containing 0, 30, 100, 300, 1,000, or 3,000 ppm p,pN-dichlorodiphenyl sulfone (equivalent to average daily doses of approximately 3.5, 15, 50, 165,or 480 mg/kg) for 14 weeks. All mice survived until the end of the study. Mean body weights of groups exposed to 300 ppm or greater were significantly less than those of the controls. Liver weights of groups exposed to 300 ppm or greater were significantly increased. Centrilobular hypertrophy of the liver was observed in most males exposed to 100 ppm or greater and in all females exposed to 1,000 or 3,000 ppm, and the severities generally increased with increasing exposure concentration. 2-YEAR STUDY IN RATS: Groups of 50 male and 50 female rats were fed diets containing 0, 10 (males), 30, 100, or 300 (females) ppm p,pN-dichlorodiphenyl sulfone for 105 weeks. Dietary concentrations of 10, 30, and 100 ppm resulted in average daily doses of approximately 0.5, 1.5, and 5.0 mg/kg to males. Dietary concentrations of 30, 100,and 300 ppm resulted in average daily doses of approximately 1.6, 5.4, and 17 mg/kg to females. Additional groups of 10 male and 10 female rats were fed the same p,pN-dichlorodiphenyl sulfone-containing diets for 18 months and bled for plasma determinations of p,pN-dichlorodiphenyl sulfone at approximately 2 weeks and 3, 12, and 18 months. Survival of all exposed groups of male and female rats was similar to that of the control groups. Mean body weights of 30 and 100 ppm males were generally less than those of the controls during the latter part of the study, and mean body weights of 100 and 300 ppm female rats were less from weeks 30 and 18,respectively. Feed consumption by the exposed groups was similar to that by the controls throughout the study. The incidences of centrilobular hepatocyte hypertrophy in 100 ppm male and 100 and 300 ppm female rats were significantly greater than those in the controls. The incidences of bile duct hyperplasia and centrilobular degeneration were also significantly increased in 100 and 300 ppm females. No neoplasms were related to chemical exposure. 2-YEAR STUDY IN MICE: Groups of 50 male and 50 female mice were fed diets containing 0, 30, 100, or 300 ppm p,pN-dichlorodiphenyl sulfone for 105 to 106 weeks. Dietary concentrations of 30, 100, and 300 ppm delivered average daily doses of approximately 4, 13, and 40 mg/kg to males and approximately 3, 10, and 33 mg/kg to females. Additional groups of 10 male and 10 female mice were fed the same p,pN-dichlorodiphenyl sulfone-containing diets for up to 12 months;three mice in each group were bled for plasma determinations of p,pN-dichloro-diphenyl sulfone at approximately 2 weeks or 3 or 12 months. Survival of all exposed groups of male and female mice was similar to that of the control groups. Mean body weights of 300 ppm mice were less than those of the controls throughout most of the study. Feed consumption by the exposed groups was similar to that by the controls throughout the study. The incidences of centrilobular hepatocyte hypertrophy in all exposed groups of male mice and in 100 and 300 ppm females were significantly greater than those in the controls. The incidence of eosinophilic foci in 300 ppm females was significantly increased. No neoplasms were related to chemical exposure. PHARMACOKINETICS OF p,pN-DICHLORODIPHENYL SULFONE: p,pN Dichlorodiphenyl sulfone is rapidly absorbed from the gut and metabolized by a saturable process. Although some p,pN-dichlorodiphenyl sulfone is eliminated unchanged in feces and urine, most of the elimination is via metabolism. Mathematical modeling of the toxicokinetics supports the view that p,pN dichlorodiphenyl sulfone induces enzymes involved in its metabolism. GENETIC TOXICOLOGY: p,pN-Dichlorodiphenyl sulfone was not mutagenic in any of several strains of Salmonella typhimurium, with or without metabolic activation enzymes (S9). Results of the sister chromatid exchange test in cultured Chinese hamster ovary cells were judged to be negative in the presence of S9 and equivocal in the absence of S9, but no induction of chromosomal aberrations was noted, with or without S9. In contrast to the in vitro results, positive results were obtained in an acute in vivo mouse bone marrow micronucleus assay with p,pN dichlorodiphenyl sulfone administered by intraperitoneal injection three times over a dose range of 200 to 800 mg/kg. CONCLUSIONS: Under the conditions of these 2-year feed studies, there was no evidence of carcinogenic activity* of p,pN dichlorodiphenyl sulfone in male F344/N rats exposed to 10, 30, or 100 ppm or in female F344/N rats exposed to 30, 100, or 300 ppm. There was no evidence of carcinogenic activity of p,pN-dichlorodiphenyl sulfone in male or female B6C3F1 mice exposed to 30,100, or 300 ppm. Exposure to p,pN-dichlorodiphenyl sulfone for 2 years caused increased incidences of nonneoplastic lesions of the liver in male and female rats and mice. PMID- 12087422 TI - Toxicology and carcinogenesis studies of indium phosphide (CAS No. 22398-90-7) in F344/N rats and B6C3F1 mice (inhalation studies). AB - Indium phosphide is used to make semiconductors,injection lasers, solar cells, photodiodes, and light-emittingdiodes. Indium phosphide was nominated for study because of its widespread use in the microelectronics industry, the potential for worker exposure,and the absence of chronic toxicity data. Male and female F344/N rats and B6C3F1 mice were exposed to indium phosphide (greater than 99% pure) by inhalation for 14 weeks or 2 years. The frequency of micronuclei was determined in the peripheral blood of mice exposed to indium phosphide for 14 weeks. 14-WEEK STUDY IN RATS: Groups of 10 male and 10 female rats were exposed to particulate aerosols of indium phosphide with amass median aerodynamic diameter of approximately 1.2 microm at concentrations of 0, 1, 3, 10, 30, or 100 mg/m3 by inhalation, 6 hours per day, 5 days per week (weeks 1 through 4 and weeks 10 through 14) or 7 days per week (weeks 5 through 9) to accommodate a concurrent teratology study. One male in the 100 mg/m3 group died before the end of the study. Body weight gains of all males and females exposed to 100 mg/m3 were less than those of the chamber controls. As a result of indium phosphide exposure, the lungs of all exposed rats had a gray to black discoloration and were significantly enlarged, weighing 2.7- to 4.4-fold more than those of the chamber controls. Indium phosphide particles were observed throughout the respiratory tract and in the lung-associated lymph nodes. A spectrum of inflammatory and proliferative lesions generally occurred in the lungs of all exposed groups of rats and consisted of alveolar proteinosis, chronic inflammation, interstitial fibrosis, and alveolar epithelial hyperplasia. Pulmonary inflammation was attended by increased leukocyte and neutrophil counts in the blood. The alveolar proteinosis was the principal apparent reason for the increase in lung weights. Indium phosphide caused inflammation at the base of the epiglottis of the larynx and hyperplasia of the bronchial and mediastinal lymph nodes. Exposure to indium phosphide affected the circulating erythroid mass. It induced a microcytic erythrocytosis consistent with bone marrow hyperplasia and hematopoietic cell proliferation of the spleen. Hepatocellular necrosis was suggested by increased serum activities of alanine aminotransferase and sorbitol dehydrogenase in all exposed groups of males and in 10 mg/m3 or greater females and was confirmed microscopically in 100 mg/m3 males and females. 14-WEEK STUDY IN MICE: Groups of 10 male and 10 female mice were exposed to particulate aerosols of indium phosphide with a mass median aerodynamic diameter of approximately 1.2 microm at concentrations of 0, 1, 3, 10, 30, or 100 mg/m3 by inhalation, 6 hours per day, 5 days per week (weeks 1 through 4 and weeks 10 through 14)or 7 days per week (weeks 5 through 9). Although the effects of indium phosphide exposure were similar in rats and mice, mice were more severely affected in that all males and females in the 100 mg/m3 groups either died or were removed moribund during the study. One male and three females in the 30 mg/m3 group were also removed before the end of the study. In general, body weight gains were significantly less in males and females exposed to 3 mg/m3 or greater compared to those of the chamber controls. Mice exposed to 30 or 100 mg/m3 were lethargic and experienced rapid, shallow breathing. As in rats, lungs were discolored and enlarged 2.6- to 4.1 fold greater than those of chamber controls due to the exposure-induced alveolar proteinosis. Indium phosphide particles were observed in the nose, trachea,larynx, and lymph nodes of some exposed males and females. Alveolar proteinosis, chronic active inflammation,interstitial fibrosis, and alveolar epithelial hyperplasia were observed; these effects were more severe than in rats. Hyperplasia in the bronchial lymph nodes and squamous metaplasia, necrosis, and suppurative inflammation of the larynx were observed in some exposed males and females. Exposure to indium phosphide induced a microcytic erythrocytosis which was consistent with the observed hematopoietic cell proliferation of the spleen.2-YEAR STUDY IN RATS Groups of 60 male and 60 female rats were exposed to particulate aerosols of indium phosphide at concentrations of 0, 0.03, 0.1, or 0.3 mg/m3, 6 hours per day,5 days per week, for 22 weeks (0.1 and 0.3 mg/m3 groups) or 105 weeks (0 and 0.03 mg/m3 groups). Animals in the 0.1 and 0.3 mg/m3 group were maintained on filtered air from exposure termination at week 22 until the end of the studies. Ten males and 10 females per group were evaluated at 3 months. 3-Month Interim Evaluation: Exposure to indium phosphide for 3 months caused a microcytic erythrocytosis and also caused enlarged lungs and lesions in the respiratory tract and lung associated lymph nodes. Although qualitatively similar to those observed in the 14-week studies, these effects were considerably less severe. However, the lesions in the lungs of rats exposed to 0.1 or 0.3 mg/m3 were considered sufficiently severe that exposure was discontinued in these groups, and the groups were allowed to continue unexposed for the remainder of the study. Survival, Body Weights, and Clinical Findings: Exposure to indium phosphide had no effect on survival or body weight gain. During the last 6 months of the study, rats in the 0.03 and 0.3 mg/m3 groups became lethargic and males breathed abnormally. Pathology Findings: At 2 years, exposure to indium phosphide caused increased incidences of alveolar/bronchiolar adenomas and carcinomas in rats. Squamous cell carcinoma of the lung occurred in four male rats exposed to 0.3 mg/m3. As observed in the 14-week study and at the 3-month interim evaluation, a spectrum of inflammatory and proliferative lesions of the lung were observed in all exposed groups of males and females;however, the extent and severity of the lesions were generally greater and included atypical hyperplasia,chronic inflammation, alveolar epithelial hyperplasia and metaplasia, alveolar proteinosis, and interstitial fibrosis. Exposure to indium phosphide also caused increased incidences of benign and malignant pheochromocytomas of the adrenal gland in males and females. Marginal increases in the incidences of mononuclear cell leukemia in males and females, fibroma of the skin in males, and carcinoma of the mammary gland in females may have been related to exposure to indium phosphide. 2-YEAR STUDY IN MICE: Groups of 60 male and 60 female mice were exposed to particulate aerosols of indium phosphide at concentrations of 0, 0.03, 0.1, or 0.3 mg/m3, 6 hours per day,5 days per week, for 21 weeks (0.1 and 0.3 mg/m3 groups) or 105 weeks (0 and 0.03 mg/m3 groups). Animals in the 0.1 and 0.3 mg/m3 groups were maintained on filtered air from exposure termination at week 21 until the end of the studies. Ten males and 10 females per group were evaluated at 3 months. 3-Month Interim Evaluation:Exposure to indium phosphide for 3 months affected the circulating erythroid mass and caused enlarged lungs and lesions in the respiratory tract and lung associated lymph nodes. These effects, although qualitatively similar to those observed in the 14-week studies, were considerably less severe. However, the lesions in the lungs of mice exposed to 0.1 mg/m3 and greater were considered sufficiently severe that exposure was discontinued in these groups and the groups were allowed to continue unexposed for the remainder of the study. Survival and Body Weights: In general, exposure to indium phosphide for 2 years reduced survival and body weight gain in exposed males and females. Pathology Findings:At 2 years, exposure to indium phosphide caused increased incidences of alveolar/bronchiolar carcinomas in males and alveolar/bronchiolar adenomas and carcinomas in females. In addition to the alveolar proteinosis and chronic active inflammation seen at earlier time points, serosa fibrosis and pleural mesothelial hyperplasia were also present. The incidences of hepatocellular neoplasms were also significantly increased in exposed males and females. Exposed groups of males and females had increased incidences of eosinophilic foci of the liver at 2 years. Marginal increases in the incidences of neoplasms of the small intestines in male mice may have been related to exposure to indium phosphide. Exposure to indium phosphide also caused inflammation of the arteries of the heart, primarily the coronary arteries and the proximal aorta, and to a lesser extent the lung-associated lymph nodes in males and in females. TISSUE BURDEN ANALYSES: Deposition and clearance studies of indium following long term exposure of rats and mice to indium phosphide by inhalation were performed. Although there were quantitative differences in lung burden and kinetic parameters for rats and mice, qualitatively they were similar. Deposition of indium in the lungs appeared to follow a zero-order (constant rate) process. Retained lung burdens throughout the studies were proportional to exposure concentration and duration. No differences in elimination rates of indium from the lungs were observed as a function of exposure concentration in either rats or mice. These studies indicated that elimination of indium was quite slow. Mice exhibited clearance half-times of 144 and 163 days for the 0.1 and 0.3 mg/m3 groups, respectively, as compared to 262 and 291 days for rats exposed to the same concentrations. The lung deposition and clearance model was used to estimate the total amount of indium deposited in the lungs of rats and mice after exposure to 0.03 mg/m3 for 2 years or to 0.1 or 0.3 mg/m3 for 21 or 22 weeks, the lung burdens at the end of the 2-year study, and the area under lung burden curves (AUC). For both species, estimates at the end of 2 years indicated that the lung burdens in the continuously exposed 0.03 mg/m3 groups were greater than those in the 0.1 or 0.3 mg/m3 groups. (ABSTRACT TRUNCATED) PMID- 12087423 TI - Genetic diversity of Nostoc microsymbionts from Gunnera tinctoria revealed by PCR STRR fingerprinting. AB - The cyanobacteria belonging to the genus Nostoc fix atmospheric nitrogen, both as free-living organisms and in symbiotic associations with a wide range of hosts, including bryophytes, gymnosperms (cycads), the small water fern Azolla (Pteridophyte), the angiosperm genus Gunnera, and fungi (lichens). The Gunnera Nostoc symbiosis is the only one that involves a flowering plant. In Chile, 12 species of Gunnera have been described with a broad distribution in the temperate region. We examined the genetic diversity of Nostoc symbionts from three populations of Gunnera tinctoria from Abtao, Chiloe Island, southern Chile, and microsymbionts from other two species of Gunnera from southern Chile, using PCR amplification of STRR (short tandemly repeated repetitive) sequences of the Nostoc infected tissue. To our knowledge, this is the first report of PCR fingerprinting obtained directly from symbiotic tissue of Gunnera. Genetic analyses revealed that Nostoc symbionts exhibit important genetic diversity among host plants, both within and between Gunnera populations. It was also found that only one Nostoc strain, or closely related strains, established symbiosis with an individual plant host. PMID- 12087424 TI - Geochemical and physiological evidence for mixed aerobic and anaerobic field biodegradation of coal tar waste by subsurface microbial communities. AB - We used geochemical analyses of groundwater and laboratory-incubated microcosms to investigate the physiological responses of naturally occurring microorganisms to coal-tar-waste constituents in a contaminated aquifer. Waters were sampled from wells along a natural hydrologic gradient extending from uncontaminated (1 well) into contaminated (3 wells) zones. Groundwater analyses determined the concentrations of carbon and energy sources (pollutants or total organic carbon), final electron acceptors (oxygen, nitrate, sulfate), and metabolic byproducts (dissolved inorganic carbon [DIC], alkalinity, methane, ferrous iron, sulfide, Mn2+). In the contaminated zone of the study site, concentrations of methane, hydrogen, alkalinity, and DIC were enhanced, while dissolved oxygen and nitrate were depleted. Field-initiated biodegradation assays using headspace-free serum bottle microcosms filled with groundwater examined metabolism of the ambient organic contaminants (naphthalene, 2-methylnaphthalene, benzothiophene, and indene) by the native microbial communities. Unamended microcosms from the contaminated zone demonstrated the simultaneous degradation of several coal-tar waste constituents at the in situ temperature (10 degrees C). Lag phases prior to the onset of biodegradation indicated the prevalence of both aerobic and anaerobic conditions in situ. Electron acceptor-amended microcosms from the most contaminated well waters demonstrated only aerobic naphthalene degradation. Collectively, the geochemical and microbial evidence show that biodegradation of coal-tar-waste constituents occurs via both aerobic and anaerobic terminal electron accepting processes at this site. PMID- 12087425 TI - Diversity of 16S rDNA and naphthalene dioxygenase genes from coal-tar-waste contaminated aquifer waters. AB - Microbial diversity in four wells along a groundwater flowpath in a coal-tar waste-contaminated aquifer was examined using RFLP analysis of both 16S rDNA and naphthalene dioxygenase (NDO) genes. Amplified ribosomal DNA restriction analysis (ARDRA) relied upon eubacteria-specific primers to generate four clone libraries. From each library, 100 clones were randomly picked for analysis. Sixty percent of 400 clones contained unique ARDRA patterns. Diversity indices calculated for each community were high (Shannon-Weaver, H = 3.53 to 3.69). Clones representing ARDRA patterns found in the highest abundance were sequenced (31 total). Sequences related to aerobic bacteria (e.g., Nitrospira, Methylomonas, and Gallionella) predominated among those retrieved from the uncontaminated area of the site, whereas sequences related to facultatively aerobic and anaerobic bacteria (e.g. Azoarcus, Syntrophus, and Desulfotomaculum) predominated among those retrieved from contaminated areas of the site. Using NDO-specific primers and low stringency PCR conditions, variability in RFLP patterns was only detected in community-derived DNA (3 of 4 wells) and not in 5 newly isolated naphthalene degrading pure cultures. The ARDRA patterns of the pure culture isolates were not found in the clone libraries. Polymorphisms in community 16S rDNA and NDO genes found in well-water microorganisms reflected distinctive geochemical conditions across the site. Sequences related to sulfate-reducing bacteria were found in groundwater that contained sulfide, while sequences related to Gallionella, Syntrophus, and nitrate-reducing aromatic hydrocarbon-degrading bacteria were found in groundwater that contained ferrous iron, methane, and naphthalene, respectively. PMID- 12087426 TI - Estimating population size and transmission bottlenecks in maternally transmitted endosymbiotic bacteria. AB - Many species of bacterial endosymbionts are acquired by animal hosts before birth, through direct transmission from mothers to eggs or embryos. This vertical transmission imposes a reduction in numbers or "bottleneck," and the size of this bottleneck affects the population structure and evolution of symbiotic lineages. We have estimated the size of the transmission bottleneck in Buchnera, the bacterial symbiont of aphids, using basic light and electron microscopy techniques. By serial-sectioning whole aphid abdomens, their eggs, and embryos, we determined the following parameters: (i) The average size of a Buchnera cell is 2.9 mm in diameter. (ii) The total number of Buchnera in an Acyrthosiphon pisum embryo was around 36700 whereas a first instar nymph contained more than 119000. (iii) The number of symbionts per bacteriocyte was around 800 in an embryo and 3200 in a first instar nymph. (iv) The total number of Buchnera transmitted to each sexual egg ranged from 850 in Nasonovia to 1800 in A. pisum to more than 8000 in Uroleucon ambrosiae. (v) The total number of secondary endosymbionts in A. pisum was 12170 for an embryo and 18360 for a first instar nymph. Secondary symbionts were arranged both extracellularly and in clusters of 2000-8000 bacteria inside bacteriocytes. These numbers are consistent with the few previous estimates of symbiont population sizes based on counts of gene copies. PMID- 12087427 TI - Distribution and characterization of Bacillus thuringiensis on the phylloplane of species of piper (Piperaceae) in three altitudinal levels. AB - Bacillus thuringiensis is found naturally on the phylloplane. In this study 35 samples from 13 species of the genus Piper (Piperaceae) were collected from three altitudinal levels located between 1800 and 2900 m above sea level in the Colombian Andean forest of Central Cordillera. Two hundred and fifty-six isolates of B. thuringiensis were obtained from 74% of the samples studied. B. thuringiensis index (number of isolates of B. thuringiensis/number of isolates of sporulated bacilli) was 0.2. The isolates were characterized by crystal morphology, the presence of cry genes by PCR, and toxicity against insects. Fifty five percent of the isolates found presented bipyramidal-crystal morphology, and 42% had round-crystal morphology. Seventy percent of the isolates amplified cry1 [cry one] genes (generally toxic to lepidopterans); 41.4% amplified cry4 and/or cry11 [cry eleven] genes (generally toxic to dipterans), and none of the isolates amplified cry3 genes (generally toxic to coleopterans). The most abundant genotype of cry genes (54.7% of the total) was cry1Aa, cry1Ab, cry1Ac, cry1Ad, and cry1B. From the total isolates found, 7.8% presented both cry1 and cry11 genes, and five isolates (2.0%) harbored cry1, cry4, and cry11 genes; all these isolates were toxic to Culex quinquefasciatus (Diptera) but not to Spodoptera frugiperda (Lepidoptera). To our knowledge, these genotypes have not been previously reported. Overall, almost 60% of the isolates were toxic to S. frugiperda, and a little more than 40% of the isolates were toxic to C. quinquefasciatus. The populations of viable vegetative cells and spores per unit area were estimated and studied statistically. No significant differences in the number of B. thuringiensis isolates per cm2 of leaf among the three altitudinal levels were found, nor were they found among the different Piper species evaluated. This study increases the knowledge of the ecology of B. thuringiensis. PMID- 12087429 TI - Exposure apportionment: ranking food items by their contribution to dietary exposure. AB - This paper identifies and ranks food items by estimating their contribution to the dietary exposure of the US population and 19 subpopulation groups. Contributions to dietary exposures to arsenic, cadmium, chromium, lead, nickel, benzene, chlorpyrifos, and diazinon are estimated using either the Dietary Exposure Potential Model (DEPM) approach, the National Human Exposure Assessment Survey Arizona (NHEXAS-AZ) approach or the combination of the two. The DEPM is a computer model that uses several national databases of food consumption and residue concentrations for estimating dietary. The DEPM approach ranks the contribution of food items to the total dietary exposure using two methods, the direct method that ranks contributions by population exposure magnitude and the weighted method that ranks by subpopulation exposure magnitude. The DEPM approach identifies highly exposed subpopulations and a relatively small number of food items contributing the most to dietary exposure. The NHEXAS-AZ approach uses the NHEXAS-AZ database containing food consumption data for each subject and chemical residues of a composite of food items consumed by each subject in 1 day during the sampling week. These data are then modeled to obtain estimates of dietary exposure to chemical residues. The third approach uses the NHEXAS-AZ consumption data with residue values from the national residue database. This approach also estimates percent contributions to exposure of each ranked food item for the Arizona population. Dietary exposures estimated using the three approaches are compared. The DEPM results indicate groups with highest dietary exposures include Nonnursing Infants, Children 1-6, Hispanic, Non-Hispanic White, Western, Northeast and Poverty 0-130%. The use of the Combined National Residue Database (CNRD) identifies 43 food items as primary contributors to total dietary exposure; they contribute a minimum of 68% of the total dietary exposure to each of the eight chemical residues. The percent contribution of ranked food items estimated using the NHEXAS samples is smaller than those obtained from the western US population via the DEPM. This indicates differences in consumption characteristics of the two groups with respect to the ranked food items. Six of 15 food items consumed by the NHEXAS-AZ subjects per day are ranked food items contributing between 56% and 70% of the estimated NHEXAS-AZ dietary exposure to each of the eight chemical residues. The difference between total dietary exposure estimates from the DEPM and NHEXAS-AZ approaches varies by chemical residue and is attributable to differences in sampling and analytical methods, and geographic areas represented by the data. Most metal exposures estimated using the NHEXAS consumption data with the CNRD have lower values than those estimated via the other approaches, possibly because the NHEXAS-AZ residue values are higher than the CNRD values. In addition, exposure estimates are seemingly affected by the difference in demographic characteristics and factors that affect types and amounts of food consumed. Efficient control strategies for reducing dietary exposure to chemical residues may be designed by focusing on the relatively small number of food items having similar ingredients that contribute substantively to the total ingestion exposure. PMID- 12087428 TI - Characterization of fecal bacterial populations in canines: effects of age, breed and dietary fiber. AB - The effects of age, breed, and diet on fecal chemistry, enzyme activity, and bacterial populations of dogs were studied. Eighteen dogs from two age groups (young: 2.5 +/- 0.5 years, old: 10.9 +/-0.7 years) and three different breeds (German shepherds, miniature schnauzers, and English setters) were rotated through a Latin Square design such that every dog was fed each of the diets. The test diets included a low-fiber (control) diet and a 10% fiber diet which contained 5% soybean hulls and 5% beet pulp. Inclusion of 10% fiber in the diet decreased the fecal concentration of ammonia, sulfide, and indole. Fiber inclusion significantly increased acetic, propionic, and butyric acid concentrations, while fecal pH decreased by 0.4 units. Fresh fecal samples were plated on selected aerobic and anaerobic culture media and DNA extracted for denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S ribosomal DNA fragments. Plate counts showed significant effects of breed (p < or = 0.05) and age (p < or = 0.01) on selected aerobic and anaerobic bacterial counts, while no significant effect of diet was found. Analysis of PCR-DGGE banding patterns showed there was a tendency for individual dogs to cluster together according to age (young or old dogs) and also for size (large or small dogs). However, the outstanding conclusion obtained from the DGGE analysis of fecal bacterial profiles was that individual dogs had their own characteristic banding pattern which was unique and stable. The relative stability and individuality of the patterns indicates that each individual harbored a characteristic fecal bacterial community which was independent of diet. PMID- 12087430 TI - A comparison of responses to a health and lifestyle questionnaire completed before and then after blood pressure screening. AB - Changes over a 2-week period in self-reported health and lifestyle information were examined and related to medical feedback provided prior to the second self report. The study group consisted of 128 females and 73 males ranging in age from 35 to 85 with a mean age of 55.5 years. On two occasions subjects completed self assessment questionnaires including 52 items covering symptomatology and various lifestyle behaviours. Before the second self-assessment, subjects were classified as cases if their blood pressure was borderline or hypertensive according to World Health Organization guidelines or as controls otherwise. Subjects were informed of this classification. Across questions, the percentage disagreement between reports ranged from 0.0% to 44.1% with a median of 17.5% and with case control differences significant at the 10% level for four questions. The mean difference between responses was significantly different from 0 at the 10% level for 15 questions with case-control differences significant at the 10% level for 6 questions. For 2 questions, the significance level of the association of response with case-control status changed from a value greater than 20% based on responses before feedback to a value less than 5% based on responses after feedback. Some evidence of recall bias was found. PMID- 12087431 TI - Mercury concentrations in urine, scalp hair, and saliva in children from Germany. AB - Mercury levels measured in urine, hair, and saliva of 245 German children (8-10 years old) are reported. Mercury concentrations in urine ranged between <0.1 and 5.3 microg/l [geometric mean (GM) 0.26 microg/l or 0.25 microg/g creatinine; median for both, 0.22 in microg/l and microg/g, respectively]. Using multiple linear regression analysis, two predictors have been found accounting for 25.3% of the variance of mercury levels in urine: the number of teeth with amalgam fillings (23.2%) and the number of defective amalgam fillings (2.1%). The mercury content in hair ranged from <0.06 to 1.7 microg/g (GM 0.18 microg/g; median 0.18 microg/g). The frequency of fish consumption, the smoking habits of the parents, and the age of the children accounted for 20.4% of the variance of mercury levels in hair. The correlation between the hair mercury content and urine mercury concentration was low (r=0.297). Mercury levels in saliva ranged between <0.32 and 4.5 microg/l (median 0.16 microg/l). The mercury concentration in saliva was below the limit of quantification of 0.32 microg/l in more than 70% of the samples. Mercury analysis in urine is suitable to estimate mercury exposure due to amalgam fillings, whereas hair mercury better reflects mercury intake by fish consumption. Up to now, saliva does not seem to be a suitable tool to monitor the mercury burden, at least not at low exposure levels. PMID- 12087433 TI - Environmental lead contamination in Miami inner-city area. AB - OBJECTIVES: The purpose of the study was to evaluate the magnitude of environmental lead contamination in the downtown area of Miami. METHODS: Lead inspections took place at 121 homes in Little Haiti and Liberty City and involved the collection of representative samples from floors, window wells, tap water, soil and air. Community health workers (CHWs) trained in interview and safety techniques went from door to door to enlist participation. On-site investigations were tailored to areas most utilized by children under the age of 6 years. The presence of lead-containing paint was also investigated in situ via X-ray fluorescence (XRF) analysis. RESULTS: Of the sampling areas, the window wells area had the most abundant occurrence of lead. On analysis, 24% of sites returned window well samples with lead levels above Department of Housing and Urban Development (HUD) guidelines. Of the soil samples, the playgrounds around the house had the highest concentration of lead. Soil sampling demonstrated that 27.5% of sites returned samples with lead levels (400 to 1600 ppm) in excess of HUD/Environmental Protection Agency (EPA) standards. Positive XRF readings in one or more components were returned by 18% of sites. CONCLUSIONS: More than half of the houses in these two neighborhoods exhibited unacceptably high levels of lead dust and soil in areas where children live and play. Limitations of this study did not allow the assessment of how many children in this area are affected. A more comprehensive study including other areas of Miami-Dade County with older housing stock is recommended. PMID- 12087432 TI - Frequency of mouthing behavior in young children. AB - Young children may be more likely than adults to be exposed to pesticides following a residential application as a result of hand- and object-to-mouth contacts in contaminated areas. However, relatively few studies have specifically evaluated mouthing behavior in children less than 5 years of age. Previously unpublished data collected by the Fred Hutchinson Cancer Research Center (FHCRC) were analyzed to assess the mouthing behavior of 72 children (37 males/35 females). Total mouthing behavior data included the daily frequency of both mouth and tongue contacts with hands, other body parts, surfaces, natural objects, and toys. Eating events were excluded. Children ranged in age from 11 to 60 months. Observations for more than 1 day were available for 78% of the children. The total data set was disaggregated by gender into five age groups (10-20, 20-30, 30 40, 40-50, 50-60 months). Statistical analyses of the data were then undertaken to determine if significant differences existed among the age/gender subgroups in the sample. A mixed effects linear model was used to test the associations among age, gender, and mouthing frequencies. Subjects were treated as random and independent, and intrasubject variability was accounted for with an autocorrelation function. Results indicated that there was no association between mouthing frequency and gender. However, a clear relationship was observed between mouthing frequency and age. Using a tree analysis, two distinct groups could be identified: children < or = 24 and children >24 months of age. Children < or = 24 months exhibited the highest frequency of mouthing behavior with 81+/-7 events/h (mean+/-SE) (n=28 subjects, 69 observations). Children >24 months exhibited the lowest frequency of mouthing behavior with 42+/-4 events/h (n=44 subjects, 117 observations). These results suggest that children are less likely to place objects into their mouths as they age. These changes in mouthing behavior as a child ages should be accounted for when assessing aggregate exposure to pesticides in the residential environment. PMID- 12087434 TI - An analysis of factors that influence personal exposure to nitrogen oxides in residents of Richmond, Virginia. AB - Nitrogen oxides (NO(x)) are ubiquitous pollutants in outdoor and indoor air. However, epidemiologic studies that evaluate health effects associated with NO(x) commonly rely upon outdoor concentrations of NO(x), nitrogen dioxide (NO(2)), or residence characteristics as surrogates for personal exposure. In this study, personal exposures (48 h) and corresponding indoor and outdoor concentrations of nitric oxide (NO), NO(2), and NO(x) were measured (July-September) in 39 adults and 9 children from 23 households in Richmond, Virginia, using Ogawa passive NO(x) monitors. Demographic, time-activity patterns, and household data were collected by questionnaire and used to develop exposure prediction models. Adults had higher NO(2), NO, and NO(x) exposures (means: 16, 63, and 79 ppb, respectively) than children (13, 49, and 62 ppb). Measurements taken in bedrooms (18, 57, and 75 ppb) and living rooms (19, 65, and 84 ppb) surpassed measurements taken outdoors (15, 21, and 36 ppb). In indoor locations, NO(x) concentrations were influenced largely by NO, and consequently, personal exposure prediction models for NO(x) were reflective of models for NO. Statistical models that best predicted personal exposures included indoor measurements; outdoor measurements contributed relatively little to personal exposure. Close to 70% of the variation in personal NO(2) and NO(x) exposure was explained by two variable models (bedroom NO(2) and time spent in other indoor locations; bedroom NO(x) and time spent in kitchen). Given appropriate resources, measurement error in epidemiologic studies can be reduced significantly with the use of personal exposure measurements or prediction models developed from indoor measurements and survey data. PMID- 12087435 TI - Estimation of environmental tobacco smoke exposure during pregnancy using a single question on household smokers versus serum cotinine. AB - Environmental tobacco smoke (ETS) exposure has been studied in relation to many diseases. The ability of a study to find an association between exposure and disease is, in part, determined by the accuracy of the exposure measure. This study examined how accurately one question, the number of smokers in the household, asked at birth, predicts ETS exposure in pregnant nonsmokers as assessed by serum cotinine. Blood specimens, drawn at 15-19 weeks gestation, from 783 women who participated in a prenatal screening program in California in 1992 were analyzed for cotinine. Serum cotinine was significantly correlated with the number of smokers in the household (r=0.35, P<0.001, geometric mean cotinine (nanograms per milliliter) for 0 smokers=0.06, 1 smoker=0.18, 2 or more smokers=0.29). Using multiple regression, the number of smokers in the household accounted for 11% of the variation in serum cotinine. Cotinine concentrations were twice as high in women living with one or more smokers compared to women not living with a smoker, when reported exposure (0 or >0 h) at home, work and other places was similar. Thus, the number of household smokers can account for a statistically significant amount of variation in serum cotinine and omission of this information would result in an underestimation of ETS exposure. Although use of this question alone does not provide an adequate estimation of ETS exposure as determined by serum cotinine, the results of this study indicate that this question is an important component of assessing ETS exposure. PMID- 12087436 TI - Continuous measurements of air change rates in an occupied house for 1 year: the effect of temperature, wind, fans, and windows. AB - A year-long investigation of air change rates in an occupied house was undertaken to establish the effects of temperature, wind velocity, use of exhaust fans, and window-opening behavior. Air change rates were calculated by periodically injecting a tracer gas (SF(6)) into the return air duct and measuring the concentration in 10 indoor locations sequentially every minute by a gas chromatograph equipped with an electron capture detector. Temperatures were also measured outdoors and in the 10 indoor locations. Relative humidity (RH) was measured outdoors and in five indoor locations every 5 min. Wind speed and direction in the horizontal plane were measured using a portable meteorological station mounted on the rooftop. Use of the thermostat-controlled attic fan was recorded automatically. Indoor temperatures increased from 21 degrees C in winter to 27 degrees C in summer. Indoor RH increased from 20% to 70% in the same time period. Windows were open only a few percent of the time in winter but more than half the time in summer. About 4600 hour-long average air change rates were calculated from the measured tracer gas decay rates. The mean (SD) rate was 0.65 (0.56) h(-1). Tracer gas decay rates in different rooms were very similar, ranging only from 0.62 to 0.67 h(-1), suggesting that conditions were well mixed throughout the year. The strongest influence on air change rates was opening windows, which could increase the rate to as much as 2 h(-1) for extended periods, and up to 3 h(-1) for short periods of a few hours. The use of the attic fan also increased air change rates by amounts up to 1 h(-1). Use of the furnace fan had no effect on air change rates. Although a clear effect of indoor-outdoor temperature difference could be discerned, its magnitude was relatively small, with a very large temperature difference of 30 degrees C (54 degrees F) accounting for an increase in the air change rate of about 0.6 h(-1). Wind speed and direction were found to have very little influence on air change rates at this house. PMID- 12087437 TI - Central giant cell granulomas: a systematic review of the radiographic characteristics with the addition of 20 new cases. AB - OBJECTIVES: The radiographic characteristics of central giant cell granuloma (CGCG) vary to a great extent. This study sought to relate the radiographic and clinical features of several cases, which were confirmed histopathologically, as central giant cell granulomas, and to systematically review the relevant literature. METHODS: Twenty cases with an established histopathological diagnosis of CGCG were received from the Department of Oral Pathology. The correlation between any two categorical variables was tested using Chi-square or Fischer's test, with alpha=0.01. A systematic review of the literature was performed using electronic and manual searches. RESULTS: There was no gender predominance. Nearly 81% of the lesions were located in the deciduous tooth bearing area of the mandible and in 35% of the cases the lesions crossed the midline. The correlation between the size of the lesion and its appearance was statistically significant with larger lesions assuming a multilocular appearance. Most lesions (55%) were radiolucent while wispy opacification and trabeculation was evident in 50% of the lesions. The borders of the lesions were well defined in 65% of the cases and scalloping was seen in 53%. Displacement of teeth and/or other anatomic structures was seen in 50% of the cases while root resorption was found in 37%. The lesions perforated the cortex in 50% of the cases. Two cases, presented with paresthesia of the lip. CONCLUSIONS: Central giant cell granuloma demonstrates clinical and radiographic features of a benign lesion with some aggressive characteristics. PMID- 12087438 TI - 3D volume rendering using multislice CT for dental implants. AB - OBJECTIVES: To determine the precision and accuracy of three-dimensional (3D) volume rendering spiral multislice computed tomography (CT)-based linear measurements of the mental foramen for dental implants, in vitro, and their precision, in vivo. METHODS: Five cadaver heads were imaged by multislice spiral CT (Toshiba Aquilion) with 0.5 mm thick axial slices (0.5 mm/0.5 s of table feed) at 0.5 mm interval reconstructions. The image data sets were transferred to a networked computer workstation. Using computer graphics the data were analysed with a 3D volume rendering technique using Vitrea software. Two oral and maxillofacial radiologists, independently, made electronic linear measurements from the superior border of the mental foramen to the crest of the alveolar process. The soft tissues were removed and physical measurements made using a 3 Space (Polhemus, Colchester, VT, USA) electromagnetic digitizer with a personal computer running Windows 98. The same linear measurements of 15 patients using the same imaging methodology were performed and the precision was analysed. RESULTS: The findings showed no statistically significant inter- or intra observer differences in vitro and in vivo, or between imaging and physical measurements in vitro (P>0.05). CONCLUSIONS: 3D multislice spiral CT imaging allows highly accurate measurements for dental implant placement in proximity to the mental foramen. Computer graphics software, using volume rendering is suitable for implant planning. PMID- 12087439 TI - Caries development in a young population managed by a restrictive attitude to radiography and operative intervention: I. A study at the patient level. AB - OBJECTIVES: To assess, in a young population (age 6-19) managed by a restrictive attitude to radiography and operative intervention, prevalence of patients with posterior caries and/or fillings, probabilities of new proximal dentine lesions given different caries experience, correlation between radiographic intervals and caries experience, and prevalence of endodontic treatment. METHODS: Bitewing radiographs, taken between age 6 through 19 in 285 patients, were analysed in succession. Occlusal and proximal surfaces were coded for caries depth (0-4) and presence of fillings (5). Simple regression analysis was used to determine relations between radiographic intervals and caries experience. Significance testings of probability estimates were made with chi(2)- and t-tests, when applicable adjusted by the Bonferroni-Holm correction for mass-significance. RESULTS: The prevalence of patients with >/ or =1 fillings/dentine lesions in occlusal surfaces increased from 6-78% and in proximal ones from 1-38%. The probability of developing new proximal dentine lesions/fillings over different time periods was significantly lower among caries-free patients than among those with enamel or dentine lesions. There was a poor correlation between radiographic intervals (mean=16 months, range=6-33) and accumulated caries experience. Endodontic treatment had been made in a total of six teeth in six patients. CONCLUSIONS: A restrictive attitude both to the frequency with which radiographs are taken and to operative treatment of proximal caries seems to be possible in young populations with low caries prevalence. PMID- 12087440 TI - Caries development in a young population managed by a restrictive attitude to radiography and operative intervention: II. A study at the surface level. AB - OBJECTIVES: To longitudinally assess the distribution of fillings and carious lesions in permanent posterior tooth surfaces, caries states fillings were made, and survival time of proximal lesions in different states in a young population managed by minimal operative treatment and a restrictive attitude to radiography. METHODS: Bitewing radiographs from age 6-20 in 285 patients were analysed. Proximal and occlusal surfaces were coded for caries depth and fillings. Start and end dates for different caries states in proximal surfaces were assigned and survival times in these states calculated by means of survival statistics. RESULTS: The frequency of carious/filled occlusal surfaces in the population increased from 6 to 19% and that of proximal surfaces from 1 to 26%. At age 19, 86% of the occlusal and 7% of the proximal surfaces affected by caries were filled. An overwhelming majority of unfilled carious proximal surfaces had enamel lesions only. Of all fillings, 81% were placed in surfaces with dentine lesions. The occlusal and proximal surfaces of the first molar and the distal surface of the second premolar accounted for most lesions and fillings among the 19-year olds. The average survival time of lesions in the enamel was 8.0 years and in the outer half of the dentine 3.4 years when right-censored data were taken into account. CONCLUSION: In a young population exposed to optimal levels of fluoride with a caries preventive programme, a restrictive attitude towards operative treatment can be combined with longer average intervals between radiographic examinations than those prescribed by current guidelines. PMID- 12087441 TI - Hand-held three-dimensional dental X-ray system: technical description and preliminary results. AB - OBJECTIVE: To design, fabricate, and perform preliminary in vitro testing of a portable, hand-held, three-dimensional (3D) dental X-ray system using options unique to Tuned-Aperture Computed Tomography (TACT). The design allows for task specific positioning over an unlimited range of user-selected angles and incorporates an integrated laser guidance system to constrain the focal-object distance. METHODS: A prototype system was fabricated consisting of an extremely lightweight X-ray source cantilevered from an aluminum gunstock-type handle to which a transparent radiation scatter shield was attached. Aiming was facilitated by task-specific visual cues used in conjunction with a spherically shaped radiolucent alignment jig coupled to the tissues of interest. Proper range was assured by a laser-assisted guidance system. The image transducer was an extrinsically modified commercially available CMOS device. The irradiated patient was simulated by a DXTTR phantom. RESULTS: The prototype demonstrated automated display of radiographed intra-oral tissues in three dimensions from arbitrary projection geometries and simulated task-specific sequences. Uncontrolled movements between exposures produced no obvious degeneration of 3D image quality. The aiming system minimized technical errors from misangulation. All dental regions in the phantom were imaged without difficulty. Projected risk was observed to be within clinically acceptable limits. CONCLUSIONS: Task-specific 3D images can be generated from as few as three uncontrolled projections. Total exposures can be prorated to levels not greatly exceeding those required for conventional two-dimensional radiographs of comparable image quality. Predicted radiation risks lie well below existing guidelines for occupational exposure. PMID- 12087442 TI - Implementation of a generalized TACT algorithm for arbitrary source-object distances. AB - OBJECTIVES: To implement, refine, and evaluate a generalized TACT reconstruction method that corrects for misregistration caused by uncontrolled variations in projective magnification, alleviates normalization artifacts at borders of backprojections, and exploits all available source information to minimize losses produced from projective truncations in three dimensions. METHODS: A new Java based software application was designed and tested in vitro using clinically representive data derived from four titanium dental implants in a cadaver jaw segment. These implants were irradiated by an intra-oral X-ray machine from various angles and distances using a solid-state sensor producing 48 radiographs. Six radiopaque markers were attached to the segment facilitating inference of associated projection geometries from analyses of the distributions of their respective shadows as seen by the sensor. Three-dimensional (3D) images were produced using the new algorithm, and the results were compared with those obtained from existing code. RESULTS: Slices processed using the new program were corrected for magnification errors. The resulting 3D displays showed significantly reduced tomosynthetic blur relative to uncorrected counterparts. The new reconstructions also minimized known border artifacts and made use of all available information. These images demonstrated apparent details otherwise hidden or lost when comparably processed using the control algorithm. CONCLUSIONS: The new software reduces both misregistration and scaling artifacts in tomosynthetically reconstructed slices. Hence, these modifications are expected to increase diagnostic accuracy and facilitate the appropriate application of TACT to an enlarged set of diagnostic tasks as compared with earlier implementations of the method. PMID- 12087443 TI - A comparison of two compression algorithms and the detection of caries. AB - OBJECTIVES: To assess the effect of two compression algorithms (JPEG and wavelet) on the detection of approximal caries. METHODS: Fifteen bitewing radiographs were generated using 100 posterior teeth mounted in blocks. The images were produced on conventional films (Ektaspeed Plus) and scanned at 300 d.p.i. Digital images were then compressed 9:1 with JPEG and wavelet methods. Nine observers detected the presence and depth of approximal caries recorded on a 5-point confidence scale and a 4-point depth scale from images viewed in random order. Histological examination provided the true depth of the lesions. Data were analysed by means of ANOVA. The null hypothesis was that there is no significant difference between the two compression algorithms and the original uncompressed images. RESULTS: JPEG performed significantly worse than the original and the wavelet algorithm (P<0.001) for the detection of dentinal lesions. However, no significant differences were found for the detection of sound surfaces, enamel lesions, and lesions up to the DEJ between JPEG-compressed images and each of the other two modalities. There was also no significant difference between the wavelet compressed images and the original for all lesion depths. CONCLUSIONS: At a compression ratio of 9:1, there were no significant differences among the original images, JPEG and wavelet compressed images for the detection of enamel caries. JPEG-compressed images performed inferiorly to the original and wavelet compressed images for the detection of dentinal lesions. Wavelet compression is a better choice than JPEG at the compression ratio investigated in this study. PMID- 12087444 TI - Image processing and enhancement provided by commercial dental software programs. AB - OBJECTIVES: To identify and analyse methods/algorithms for image processing provided by various commercial software programs used in direct digital dental imaging and to map them onto a standardized nomenclature. METHODS: Twelve programs presented at the 28th International Dental-Show, March, 2001, Cologne, Germany and the Emago advanced software were included in this study. An artificial test image, comprised of gray scale ramps, step wedges, fields with Gaussian-distributed noise, and salt and pepper noise, was synthesized and imported to all programs to classify algorithms for display; linear, non-linear and histogram-based point processing; pseudo-coloration; linear and non-linear spatial filtering; frequency domain filtering; measurements; image analysis; and annotations. RESULTS: The 13 programs were found to possess a great variety of image processing and enhancement facilities. All programs offer gray-scale image display with interactive brightness and contrast adjustment and gray-scale inversion as well as calibration and length measurements. While Emago enables arbitrary spatial filtering with user-defined masks up to 7x7 pixels in size, most programs sparsely include filters and tools for image analysis and comparison. Moreover, the naming and implementation of provided functions differ. Some functions inappropriately use standardized image processing terms to describe their operations. CONCLUSIONS: Image processing and enhancement functions are rarely incorporated in commercial software for direct digital imaging in dental radiology. Until now, comparison of software was limited by the arbitrary naming used in each system. Standardized terminology and increased functionality of image processing should be offered to the dental profession. PMID- 12087445 TI - Accuracy of reformatted CT image for measuring the pre-implant site: analysis of the image distortion related to the gantry angle change. AB - OBJECTIVES: In a multiplanar reconstruction CT program (CT/MPR), the altered gantry angle corresponding to the patient's position during scanning for multiplanar reformatted CT may cause distortion of the image. The aim of this study was to quantitatively assess the distortion in reformatted central panoramic and cross-sectional images owing to the change of gantry angle. METHODS: A resin block model with four cylindrical holes and a human dry mandible were used in high resolution conventional CT scanning. A MPR software package was used for reformatted panoramic and cross-sectional images. The block and the gantry were equally inclined at 0 degrees, 15 degrees and 30 degrees. The relationship between the positional difference and depth of the hole as determined in the reformatted image, and the actual depth was analysed with respect to the gantry angle. The actual depths (H) were calculated by use of the measured depths (h) and the inclined angles: H=hxcos beta. The positional differences of the hole bases (W) were also calculated by use of the measured depths and the inclined angles: W=hxsin beta or Hxtan beta. RESULTS: Increasing the gantry angle, increased the depth and inclination of the holes in the reformatted central panoramic images. Inclined angle (beta) was the same as gantry angle. CONCLUSION: When the gantry angle is not at 0 degrees, certain CT/MPR programs may distort the reformatted image. If distortion occurs, the corrected position and usable length of pre-implant sites can be calculated by use of the above formula. PMID- 12087446 TI - Effect of over- and underexposure on the sharpness of the image of a marker in computer-assisted dental implant tomography. AB - In dental implant tomography, a clinician typically makes several tomograms of cross-sectional and sagittal slices at and near the intended implantation site. The slice with the sharpest image of the metal marker is deemed to be the correct implantation site; the other slices with blurred images of the marker are those made either mesially or distally to the marker. However, if the images were over- or underexposed, the marker will be blurred on all the slices and a dentist may be wrongly accused of having placed a dental implant at an improper site with possible medico-legal ramifications. PMID- 12087447 TI - Why hasn't the National Institute been 'NICE' to patients with colorectal cancer? National Institute of Clinical Excellence. PMID- 12087448 TI - Which endpoints should we use in evaluating the use of novel fluoropyrimidine regimens in colorectal cancer? AB - Although significant advances have been made in the treatment of advanced/metastatic colorectal cancer, 5-fluorouracil (5-FU) still forms the basis of chemotherapy. Recently, new 5-FU schedules and novel fluoropyrimidines have been developed, but there are no trials directly comparing these regimens. The current review describes the mechanisms of action, pre-clinical and phase I/II studies of two oral fluoropyrimidine therapies, capecitabine and uracil with tegafur plus leucovorin. It also compares the phase III studies of these agents with those of the two most popular infusional 5-FU-based regimens: de Gramont and German AIO (The Association of Medical Oncology (AIO) of the German Cancer Society). Both oral and infusional regimens demonstrated similar survival to the Mayo Clinic regimen, a standard treatment for colorectal cancer. Therefore, other endpoints must be examined to decide optimum therapy, including response rates, time to disease progression, tolerability and patients' convenience. All four new therapies demonstrated superior safety profiles compared with the Mayo Clinic regimen. However the uracil with tegafur plus leucovorin regimen was associated with severe diarrhoea and capecitabine with hand-foot syndrome. Patients will not sacrifice efficacy for the convenience of oral therapy alone, therefore the fact that capecitabine achieved superior response rates and equivalent time to disease progression compared with the Mayo Clinic regimen, while uracil with tegafur plus leucovorin produced lower response rates and significantly inferior time to disease progression, is highly relevant in choosing treatment. PMID- 12087449 TI - Clinical and economic benefits of irinotecan in combination with 5-fluorouracil and folinic acid as first line treatment of metastatic colorectal cancer. AB - The combination of irinotecan plus 5-fluorouracil and folinic acid has clinical and survival benefits over 5-fluorouracil and folinic acid alone in the setting of first line treatment of metastatic colorectal cancer. The aim of this cost effectiveness analysis was to compare the economic implications, from a UK health commissioner perspective, of the two treatment arms (de Gramont regimen) in this setting. Resource utilisation data collected prospectively during the study were used as a basis for estimating cumulative drug dosage, chemotherapy administration, and treatment of complications during first line therapy. Resource utilisation associated with further chemotherapy in patients who had progressed during the study was derived from a retrospective case note review. Drug acquisition costs were derived from the British National Formulary (September, 2001) and unit costs for clinical consultation and services were taken from the latest relevant cost database. Cumulative costs per patient associated with further chemotherapy were lower in the irinotecan plus 5 fluorouracil and folinic acid treatment arm. Based on incremental costs per life year gained of 14 794 pounds sterling, the combination of irinotecan plus 5 fluorouracil and folinic acid can be considered cost-effective by commonly accepted criteria compared with 5-fluorouracil and folinic acid alone. Thus, clinical and economic data demonstrate that irinotecan, either in combination with irinotecan plus 5-fluorouracil and folinic acid in the first line setting or as monotherapy in the second line setting, has a major role in the management of metastatic colorectal cancer. PMID- 12087450 TI - Costs and consequences of different chemotherapy regimens in metastatic colorectal cancer. AB - An economic sub-study was run alongside a large multi-centre randomised trial (MRC-CR06) comparing three chemotherapy regimens; de Gramont, Lokich and raltitrexed in patients with metastatic colorectal cancer. Patients in six of 45 centres in the main trial were approached to take part in the sub-study. Chemotherapy delivery costs were assessed in each sub-study centre with external validity verified by questionnaire to all other centres. Patient representativeness was assessed. Stochastic resource use data, including patient borne costs and non-hospital health service resource use were monitored prospectively. Mean total societal costs were de Gramont= 5051 pounds sterling (s.d. 1910 pounds sterling ), raltitrexed= 2616 pounds sterling (s.d. 991 pounds sterling ) and Lokich= 2576 pounds sterling (s.d. 1711 pounds sterling ). In pairwise comparisons, statistically significant mean total cost differences were shown for de Gramont vs Lokich (mean difference= 2475 pounds sterling , 95%CI 914 pounds sterling - 4037 pounds sterling , P<0.01) and for de Gramont vs raltitrexed (mean difference= 2435 pounds sterling, 95%CI 922 pounds sterling - 2948 pounds sterling , P<0.01). Sensitivity analyses showed little effect on overall costs. The main trial showed de Gramont and Lokich to be equally effective in terms of survival, quality of life and response rates but Lokich had higher toxicity and hand-foot syndrome. Raltitrexed showed similar response rates and overall survival but increased toxicity and inferior quality of life making it a clinically inferior regimen despite its ease of administration and costs. For a comparable clinical outcome, Lokich can be administered for approximately half the cost of de Gramont. PMID- 12087451 TI - Serum tryptophan decrease correlates with immune activation and impaired quality of life in colorectal cancer. AB - Cancer-related indoleamine (2,3)-dioxygenase up-regulation by interferon-gamma might influence quality of life by depleting serum tryptophan. We correlated serum tryptophan levels with immune activation and quality of life in patients with colorectal liver metastases. Venous blood was sampled from patients with primary colorectal cancer and from patients with metachronous colorectal liver metastases who completed quality of life and psychological questionnaires. Serum tryptophan, kynurenine, neopterin, interleukin 2 soluble receptor alpha (IL-2 sRalpha), soluble tumour necrosis factor receptor I (sTNF RI), interleukin 6, and C-reactive protein were measured. Liver metastasis volume was estimated by computerised tomography, and survival from blood sampling was noted. Sixty-six patients with colorectal cancer were studied (39 males; median age 66 years) of whom 25 had colorectal liver metastases only (17 males; median age 62 years; median liver metastasis volume 208 ml; median survival 234 days). Reduced serum tryptophan was significantly associated with Rotterdam Symptom Checklist physical symptom (r=-0.51, P=0.01) and Sickness Impact Profile (r=-0.42, P=0.04) scores, and correlated with increased serum neopterin (r=-0.36, P=0.003), IL-2 sRalpha (r=-0.51, P=0.01) and sTNF RI (r=-0.45, P=0.02) levels. Stepwise regression analyses suggested that serum tryptophan was an independent predictor of Rotterdam Symptom Checklist physical symptom (regression coefficient -20.78, P=0.01) and Sickness Impact Profile (regression coefficient -109.09, P=0.04) scores. The results supported a role for interferon-gamma-mediated serum tryptophan decrease in cancer-induced quality of life deterioration. PMID- 12087452 TI - Early decline in left ventricular ejection fraction predicts doxorubicin cardiotoxicity in lymphoma patients. AB - Thirty adult patients with non-Hodgkin's lymphoma were studied to evaluate prospectively the significance of early decline in left ventricular ejection fraction after low cumulative doxorubicin dose (200 mg x m(-2)) in predicting the later impairment of left ventricular function. Cardiac function was monitored with radionuclide ventriculography at baseline and after cumulative doxorubicin doses of 200, 400 and 500 mg x m(-2). Cardiotoxicity was defined as a decrease in left ventricular ejection fraction of more than 10% units to a final left ventricular ejection fraction < or =50%. Twenty-eight patients received doxorubicin > or =400 mg x m(-2) and were evaluable for cardiotoxicity. Clinical heart failure developed in two patients (7%) after a cumulative doxorubicin dose of 500 mg m(-2). Left ventricular ejection fraction decreased more than 10% absolute ejection fraction units to a final left ventricular ejection fraction < or =50% in 10 patients (36%). Left ventricular ejection fraction decreased from 56+/-1.5% to 53.6+/-1.5% (P=0.016) in patients with no cardiotoxicity, and from 60.8+/-2.4% to 41.8+/-2.0% (P<0.001) in patients with cardiotoxicity. For patients who developed cardiotoxicity, the fall in left ventricular ejection fraction after a cumulative doxorubicin dose of only 200 mg x m(-2) was highly significant (left ventricular ejection fraction 49.7+/-1.8%, P=0.001 vs baseline). In receiver operator characteristic analysis, the area under the curve for the decrease in left ventricular ejection fraction at a cumulative doxorubicin dose of 200 mg x m(-2) for predicting cardiotoxicity in all patients was 0.858. The decrease in left ventricular ejection fraction of more than 4% units after a cumulative doxorubicin dose of 200 mg x m(-2) had a 90% sensitivity and 72% specificity for predicting later cardiotoxicity. Our results show that the significant impairment of left ventricular function during doxorubicin therapy can be predicted early, already at low cumulative doxorubicin doses. This finding may be of value in identifying patients at high or low risk for the development of anthracycline cardiotoxicity. PMID- 12087453 TI - A phase I study of combination chemotherapy with gemcitabine and oral UFT for advanced non-small cell lung cancer. AB - A phase I study was carried out to determine the optimal dose and administration schedule for combined UFT plus gemcitabine therapy in patients with non-small cell lung cancer. Twenty-four patients (including 11 patients previously treated with cisplatin as the key drug) received oral UFT 400 mg x m(-2) on days 1 to 14 with intravenous infusions of gemcitabine (800 mg x m(-2) on days 8 and 15, or 900 mg x m(-2) on days 8 and 15, or 900 mg x m(-2) on days 1, 8 and 15). The most appropriate dosing option appeared to be 400 mg x m(-2) per day of oral UFT for 14 consecutive days with 900 mg x m(-2) gemcitabine on days 8 and 15. Eight of the 24 patients achieved partial response. The combination chemotherapy UFT and gemcitabine was well tolerated and may benefit patients with advanced non-small cell lung cancer. A multicentre phase II study using a 3-weekly regimen is in progress. PMID- 12087454 TI - Duration of adjuvant chemotherapy for breast cancer: a joint analysis of two randomised trials investigating three versus six courses of CMF. AB - Cyclophosphamide, methotrexate and fluorouracil adjuvant combination chemotherapy for breast cancer is currently used for the duration of six monthly courses. We performed a joint analysis of two studies on the duration of adjuvant cyclophosphamide, methotrexate and fluorouracil in patients with node-positive breast cancer to investigate whether three courses of cyclophosphamide, methotrexate and fluorouracil might suffice. The International Breast Cancer Study Group Trial VI randomly assigned 735 pre- and perimenopausal patients to receive 'classical' cyclophosphamide, methotrexate and fluorouracil for three consecutive cycles, or the same chemotherapy for six consecutive cycles. The German Breast Cancer Study Group randomised 289 patients to receive either three or six cycles of i.v. cyclophosphamide, methotrexate and fluorouracil day 1, 8. Treatment effects were estimated using Cox regression analysis stratified by clinical trial without further adjustment for covariates. The 5-year disease-free survival per cents (+/-s.e.) were 54+/-2% for three cycles and 55+/-2% for six cycles (n=1024; risk ratio (risk ratio: CMFx3/CMFx6), 1.00; 95% confidence interval, 0.85 to 1.18; P=0.99). Use of three rather than six cycles was demonstrated to be adequate in both studies for patients at least 40-years-old with oestrogen-receptor-positive tumours (n=594; risk ratio, 0.86; 95% confidence interval, 0.68 to 1.08; P=0.19). In fact, results slightly favoured three cycles over six for this subgroup, and the 95% confidence interval excluded an adverse effect of more than 2% with respect to absolute 5-year survival. In contrast, three cycles appeared to be possibly inferior to six cycles for women less than 40-years-old (n=190; risk ratio, 1.25; 95% confidence interval, 0.87 to 1.80; P=0.22) and for women with oestrogen-receptor-negative tumours (n=302; risk ratio, 1.15; 95% confidence interval, 0.85 to 1.57; P=0.37). Thus, three initial cycles of adjuvant cyclophosphamide, methotrexate and fluorouracil chemotherapy were as effective as six cycles for older patients (40-years-old) with oestrogen receptor-positive tumours, while six cycles of adjuvant cyclophosphamide, methotrexate and fluorouracil might still be required for other cohorts. Because endocrine therapy with tamoxifen and GnRH analogues is now available for younger women with oestrogen-receptor-positive tumours, the need for six cycles of cyclophosphamide, methotrexate and fluorouracil is unclear and requires further investigation. PMID- 12087455 TI - High activity Rhenium-186 HEDP with autologous peripheral blood stem cell rescue: a phase I study in progressive hormone refractory prostate cancer metastatic to bone. AB - We tested the feasibility and toxicity of high activities Rhenium-186 hydroxyethylidene diphosphonate, with peripheral blood stem cell rescue in patients with progressive hormone refractory prostate cancer metastatic to bone. Twenty-five patients received between 2500 and 5000 MBq of Rhenium-186 hydroxyethylidene diphosphonate followed 14 days later by the return of peripheral blood peripheral blood stem cells. Activity limiting toxicity was defined as grade III haematological toxicity, lasting at least 7 days, or grade IV haematological toxicity of any duration or any serious unexpected toxicity. Activity limiting toxicity occurred in two of six who received activities of 5000 MBq and maximum tolerated activity was defined at this activity level. Prostate specific antigen reductions of 50% or more lasting at least 4 weeks were seen in five of the 25 patients (20%) all of whom received more than 3500 MBq of Rhenium 186 hydroxyethylidene diphosphonate. The actuarial survival at 1 year is 54%. Administered activities of 5000 MBq of Rhenium-186 hydroxyethylidene diphosphonate are feasible using autologous peripheral blood peripheral blood stem cell rescue in patients with progressive hormone refractory prostate cancer metastatic to bone. The main toxicity is thrombocytopaenia, which is short lasting. A statistically significant activity/prostate specific antigen response was seen. We have now commenced a Phase II trial to further evaluate response rates. PMID- 12087456 TI - The United Kingdom Childhood Cancer Study of exposure to domestic sources of ionising radiation: 1: radon gas. AB - This paper reports the results of the United Kingdom Childhood Cancer Study relating to risks associated with radon concentrations in participants homes at the time of diagnosis of cancer and for at least 6 months before. Results are given for 2226 case and 3773 control homes. No evidence to support an association between higher radon concentrations and risk of any of the childhood cancers was found. Indeed, evidence of decreasing cancer risks with increasing radon concentrations was observed. Adjustment for deprivation score for area of residence made little difference to this trend and similar patterns were evident in all regions and in all diagnostic groups. The study suggests that control houses had more features, such as double glazing and central heating, leading to higher radon levels than case houses. Further, case houses have features more likely to lead to lower radon levels, e.g. living-rooms above ground level. Consequently the case-control differences could have arisen because of differences between houses associated with deprivation that are not adequately allowed for by the deprivation score. PMID- 12087457 TI - The United Kingdom Childhood Cancer Study of exposure to domestic sources of ionising radiation: 2: gamma radiation. AB - This article reports measurements of household levels of gamma and cosmic rays at the addresses of children with cancer at the time of diagnosis and six months before, and of similar data at the addresses of control children. There is no indication of increased risk with increasing dose rates either in matched or unmatched analyses, with or without adjustment for deprivation. Sub-division by diagnostic group did not reveal any association with any specific types of malignancy. Studies of the relationship between household gamma rays and radon concentration show no evidence of any interactions. PMID- 12087458 TI - Cancer risks in populations living near landfill sites in Great Britain. AB - Previous studies have raised concerns about possible excess risks of bladder, brain and hepatobiliary cancers and leukaemias near landfill sites. Several cancers have been implicated, but no consistent pattern has emerged. We present a large nationwide analysis of selected cancers near landfill sites in Great Britain. The base population comprised people living within 2 km of 9565 (from a total of 19 196) landfill sites that were operational at some time from 1982 to 1997, with populations living more than 2 km from a landfill as reference. Risks of cancers at the above sites were computed with adjustment for age, sex, year of diagnosis, region and deprivation. National post-coded registers provided a total of 341 856 640 person-years for the adult cancer analyses and 113 631 443 person years for childhood leukaemia. There were 89 786 cases of bladder cancer, 36 802 cases of brain cancer, 21 773 cases of hepatobiliary cancer, 37 812 cases of adult leukaemia and 3973 cases of childhood leukaemia. In spite of the very large scale of this national study, we found no excess risks of cancers of the bladder and brain, hepatobiliary cancer or leukaemia, in populations living within 2 km of landfill sites. The results were similar if the analysis were restricted to landfill sites licensed to carry special (hazardous) waste. Our results do not support suggestions of excess risks of cancer associated with landfill sites reported in other studies. PMID- 12087459 TI - Cancer risks among long-standing spouses. AB - We estimated risks for concordant and discordant cancers in spouses in order to quantify cancer risks from the shared environment. The study was restricted to spouses who had one or more children in common and who lived together for at least 15 years after the first child's birth. The nation-wide Family-Cancer Database was used as the source of family and cancer data. Standardised incidence ratios were calculated for concordant and discordant cancers in spouses after 50 years of age. Among the 18 cancer sites considered, only three sites, stomach, lung and bladder, showed concordant increases of cancer among spouses, standardised incidence ratios ranging only from 1.19 to 1.38. Additionally, gastric and pancreatic cancer were associated among spouses, as did many cancers which were related to tobacco smoking or human papilloma virus infection. By contrast, standardised incidence ratios of colon, rectal, renal and skin cancers showed no increases among spouses. Shared lifestyle among family members seems to explain only a small proportion of familial cancer susceptibility. Because lifestyles are likely to differ more between parents and offspring than between spouses, familial cancer risks between parents and offspring are even more likely to be due to heritable than environmental effects. PMID- 12087460 TI - A prospective study of serum bile acid concentrations and colorectal cancer risk in post-menopausal women on the island of Guernsey. AB - Secondary bile acids produced by the action of the colonic microflora may increase risk of colorectal cancer. Serum bile acid concentrations reflect the faecal bile acid profile and may be of value as biomarkers of risk of colorectal cancer. In a pilot investigation we examined: (i) the reproducibility of measurements of serum bile acids in two blood samples collected several years apart; and (ii) the hypothesis that relatively high levels of secondary bile acids, particularly deoxycholic acid, would be positively associated with an increased risk of colorectal cancer in a prospective study of 3680 women in Guernsey. There was poor reproducibility between repeat measurements of absolute serum concentrations of bile acids, but there was moderately good reproducibility for the ratios of serum concentrations of deoxycholic/cholic acid, lithocholic/chenodeoxycholic and secondary/primary bile acid concentrations (duplicate blood samples were available for 30 women). There were no significant differences in ratios of serum secondary to primary bile acids or in absolute concentrations of bile acids between the 46 women who developed colorectal cancer and their matched controls, although there was a suggestion that an increased risk was associated with a high ratio of deoxycholic/cholic acid (relative risk in top third compared to lower third=3.92 (95% CI 0.91-17.0, P for trend=0.096). These findings suggest that the ratios of serum bile acid concentrations are sufficiently reproducible for epidemiological studies, but that a larger study than our own is needed to adequately test the hypothesis of their relation to cancer risk. PMID- 12087461 TI - Benign thyroid disease and dietary factors in thyroid cancer: a case-control study in Kuwait. AB - We conducted a population-based study of 313 case-control pairs in Kuwait to examine the aetiology of thyroid cancer, the second most common neoplasm among women in this and several other countries in the Gulf region. Among the demographic variables, individuals with 12+ years of education had a significantly reduced risk of thyroid cancer (OR=0.6; 95% CI: 0.3-0.9). The average age at diagnosis (+/-s.d.) of thyroid cancer was 34.7+/-11 years in women and 39+/-13.4 years in men. History of thyroid nodule was reported only by cases (n=34; 10.9%; lower 95% CI: 12.0); and goitre by 21 cases and four controls (OR=5.3; 95% CI: 1.8-15.3). There was no significant increase in risk with history of hypothyroidism (OR=1.8) or hyperthyroidism (OR=1.7). For any benign thyroid disease, the OR was 6.4 (95% CI: 3.4-12.0); and the population attributable risk was about 26% (95% CI: 21.1-30.9). Stepwise regression analysis showed that high consumption of processed fish products (OR=2.2; 95% CI: 1.6-3.0) fresh fish (OR=0.5; 95% CI: 0.4-0.7) and chicken (OR=1.7; 95% CI: 1.2-2.3) were independently associated with thyroid cancer with significant dose-response relationships. Among the thyroid cancer patients who reported high consumption of fish products, a large majority also reported high consumption of fresh fish (98%) and shellfish (68%). No clear association emerged with consumption of cruciferous vegetables. These data support the hypothesis that hyperplastic thyroid disease is strongly related to thyroid cancer; and that habitual high consumption of various seafoods may be relevant to the aetiology of thyroid cancer. The association with chicken consumption requires further study. PMID- 12087462 TI - Risk factors for oesophageal, lung, oral and laryngeal cancers in black South Africans. AB - The authors used data collected from 1995 to 1999, from an on-going cancer case control study in greater Johannesburg, to estimate the importance of tobacco and alcohol consumption and other suspected risk factors with respect to cancer of the oesophagus (267 men and 138 women), lung (105 men and 41 women), oral cavity (87 men and 37 women), and larynx (51 men). Cancers not associated with tobacco or alcohol consumption were used as controls (804 men and 1370 women). Tobacco smoking was found to be the major risk factor for all of these cancers with odds ratios ranging from 2.6 (95% CI 1.5-4.5) for oesophageal cancer in female ex smokers to 50.9 (95% CI 12.6-204.6) for lung cancer in women, and 23.9 (95% CI 9.5-60.3) for lung cancer and 23.6 (95% CI 4.6-121.2) for laryngeal cancer in men who smoked 15 or more grams of tobacco a day. This is the first time an association between smoking and oral and laryngeal cancers has been shown in sub Saharan Africa. Long-term residence in the Transkei region in the southeast of the country continues to be a risk factor for oesophageal cancer, especially in women (odds ratio=14.7, 95% CI 4.7-46.0), possibly due to nutritional factors. There was a slight increase in lung cancer (odds ratio=2.9, 95% CI 1.1-7.5) in men working in 'potentially noxious' industries. 'Frequent' alcohol consumption, on its own, caused a marginally elevated risk for oesophageal cancer (odds ratio=1.7, 95% CI 1.0-2.9, for women and odds ratio=1.8, 95% CI 1.2-2.8, for men). The risks for oesophageal cancer in relation to alcohol consumption increased significantly in male and female smokers (odds ratio=4.7, 95% CI=2.8 7.9 in males and odds ratio=4.8, 95% CI 3.2-6.1 in females). The above results are broadly in line with international findings. PMID- 12087463 TI - Frequent expression of new cancer/testis gene D40/AF15q14 in lung cancers of smokers. AB - We found a significant correlation between lung cancer in smokers and the expression of a human gene, D40, predominantly expressed in testis and cancers. In an attempt to clone a novel human gene, we screened a cDNA library derived from a human B cell line and obtained a cDNA clone that we refer to as D40. A search for public databases for sequence homologies showed that the D40 gene is identical to AF15q14. D40 mRNA is predominantly expressed in normal testis tissue. However, this gene is also expressed in various human tumour cell lines and primary tumours derived from various organs and tissues, such as lung cancer. We examined the relationship between D40 expression and clinico-pathological characteristics of tumours in primary lung cancer. D40 expression did not significantly correlate with either histological type or pathological tumour stage. However, D40 expression was observed more frequently in poorly differentiated tumours than in well or moderately differentiated ones. Furthermore, the incidence of D40 expression was significantly higher in tumours from patients who smoke than in those from non-smokers. D40/AF15q14 is the first gene in the cancer/testis family for which expression is related to the smoking habits of cancer patients. PMID- 12087464 TI - Prognostic impact of chromosomal aberrations in Ewing tumours. AB - Although greater than 50% of Ewing tumours contain non-random cytogenetic aberrations in addition to the pathognomonic 22q12 rearrangements, little is known about their prognostic significance. To address this question, tumour samples from 134 Ewing tumour patients were analysed using a combination of classical cytogenetics, comparative genomic and fluorescence in situ hybridisation. The evaluation of the compiled data revealed that gain of chromosome 8 occurred in 52% of Ewing tumours but was not a predictive factor for outcome. Gain of 1q was associated with adverse overall survival and event-free survival in all patients, irrespective of whether the tumour was localised or disseminated (overall survival: P=0.002 and P=0.029; event-free survival: P=0.018 and P=0.010). Loss of 16q was a significant predictive factor for adverse overall survival in all patients (P=0.008) and was associated with disseminated disease at diagnosis (P=0.039). Gain of chromosome 12 was associated with adverse event free survival (P=0.009) in patients with localised disease. These results indicate that in addition to a 22q12 rearrangement confirmation in Ewing tumours it is important to assess the copy number of 1q and 16q to identify patients with a higher probability of adverse outcome. PMID- 12087465 TI - Expression of basic fibroblast growth factor is associated with poor outcome in non-Hodgkin's lymphoma. AB - It is now clear that angiogenesis and angiogenesis factors are important in the pathogenesis of haematological malignancies. High pretreatment levels of serum basic fibroblast growth factor have been shown to be associated with poor prognosis in patients with non-Hodgkin's lymphoma. The aim of this study was to evaluate whether non-Hodgkin's lymphoma cells express basic fibroblast growth factor and/or its receptor (fibroblast growth factor receptor-1) and whether basic fibroblast growth factor expression correlates with basic fibroblast growth factor serum levels, intratumoral microvessel density, and patient outcome. We measured basic fibroblast growth factor by enzyme-linked immunosorbent assay in sera taken from 58 patients with non-Hodgkin's lymphoma before treatment and in 19 of them also after treatment. Pathological specimens at diagnosis were evaluated by immunohistochemistry staining using polyoclonal antibody against factor-VIII-related antigen, basic fibroblast growth factor and fibroblast growth factor receptor-1 to determine the expression of the microvessel count and basic fibroblast growth factor and fibroblast growth factor receptor-1. The lymphoma specimens demonstrated positive staining for basic fibroblast growth factor (in 23%) and fibroblast growth factor receptor-1 (in 58.5%). The patients who expressed basic fibroblast growth factor had a significantly worse progression free and overall survival than those who did not (P=0.003 and P=0.03 respectively), while patients expressing fibroblast growth factor receptor-1 were less likely to achieve complete remission than those lacking the receptor (33% vs 65%, P=0.047). There was no correlation of basic fibroblast growth factor staining with either serum basic fibroblast growth factor levels or microvessel count. Basic fibroblast growth factor serum levels did not change significantly after treatment These results suggest that non-Hodgkin's lymphoma specimens express basic fibroblast growth factor and its receptor (fibroblast growth factor receptor-1) and this expression is associated with poor patient outcome. PMID- 12087467 TI - A comparison of serum and plasma levels of vascular endothelial growth factor during the menstrual cycle in healthy female volunteers. AB - Angiogenesis is the formation of new blood vessels from the existing vasculature, and is essential for the growth and metastasis of most solid tumours. One of the most important growth factors involved in the angiogenesis process is vascular endothelial growth factor. Vascular endothelial growth factor expression has been shown to be regulated by female hormones in breast cancer cell lines, and two previous authors have reported on cyclical variations in serum vascular endothelial growth factor concentrations with conflicting results. No work has been performed on variations in plasma levels of vascular endothelial growth factor during the menstrual cycle. We therefore conducted the first prospective trial to compare serum and plasma levels of vascular endothelial growth factor in healthy pre-menopausal volunteers. Twenty healthy pre-menopausal women were recruited and had blood samples taken over one menstrual cycle with an average of eight samples taken per patient. Plasma and serum samples were then analysed for sex hormones and vascular endothelial growth factor 165. Serum vascular endothelial growth factor levels were found to be significantly higher than plasma vascular endothelial growth factor levels (P<0.005). We found no significant difference between serum and plasma vascular endothelial growth factor in the luteal and follicular phases of the cycle. The majority of the measurements for plasma levels of vascular endothelial growth factor at all phases of the cycle were under the limit of detection of the vascular endothelial growth factor ELISA kit. We found no significant correlation between plasma or serum levels of vascular endothelial growth factor and either FSH, LH, Oestradiol or Progesterone levels. This study has demonstrated no difference in serum concentrations of vascular endothelial growth factor during the different phases of the menstrual cycle in a group of healthy volunteers. We also demonstrated no obvious difference in plasma concentrations of vascular endothelial growth factor between the phases of the cycle, but most of the measurements were below the level of accuracy reported by the ELISA kit manufacturer. With the sensitivity of this ELISA test, therefore, we must still regard the question of whether there is a variation in plasma concentrations of vascular endothelial growth factor throughout the menstrual cycle as unanswered. PMID- 12087466 TI - CD10 is a marker for cycling cells with propensity to apoptosis in childhood ALL. AB - CD10 constitutes a favourable prognostic marker for childhood acute lymphoblastic leukaemia. Since correlations between CD10, cell cycle and apoptotic abilities were demonstrated in various cell types, we investigated whether differences existed in the cycling/apoptotic abilities of CD10-positive and CD10-negative B acute lymphoblastic leukaemia cells. Twenty-eight cases of childhood acute lymphoblastic leukaemia (mean age of 6.8 years) were subdivided into two groups according to high (17 cases, 93.2+/-4.5%, MRFI 211+/-82 CD10-positive cells) or low (11 cases, 11.5+/-6.2%, MRFI 10+/-7 CD10-negative cells) expression of CD10. CD10-positive acute lymphoblastic leukaemia cells were cycling cells with elevated c-myc levels and propensity to apoptosis, whereas CD10-negative acute lymphoblastic leukaemia cells had lower cycling capacities and c-myc levels, and were resistant to apoptosis in vitro. A close correlation between all these properties was demonstrated by the observations that the few CD10-positive cells found in the CD10-negative acute lymphoblastic leukaemia group displayed elevated c-myc and cycling capacities and were apoptosis prone. Moreover, exposure of CD10 positive acute lymphoblastic leukaemia B cells to a peptide nucleic acid anti gene specific for the second exon of c-myc caused inhibition of c-myc expression and reduced cell cycling and apoptotic abilities as well as decreased CD10 expression. PMID- 12087468 TI - Higher expression of human kallikrein 10 in breast cancer tissue predicts tamoxifen resistance. AB - The human tissue kallikreins are secreted serine proteases, encoded by a group of homologous genes clustered in tandem on chromosome 19q13.3-4. Human kallikrein 6 and human kallikrein 10 are two new members of this family. Recently, we developed highly sensitive and specific immunofluorometric assays for human kallikrein 6 and human kallikrein 10, which allow for their quantification in tissue extracts and biological fluids. Both human kallikrein 6 and human kallikrein 10 are found to be down-regulated in breast cancer cell lines, suggesting that they may be involved in breast cancer pathogenesis and progression. In this study, we investigated the potential value of human kallikrein 6 and human kallikrein 10 as prognostic and predictive factors in breast cancer. We quantified human kallikrein 6 and human kallikrein 10 protein levels in 749 breast tumour cytosolic extracts and correlated this data with various clinicopathological variables and patient outcomes. Human kallikrein 6 and human kallikrein 10 are positively correlated with each other. Higher human kallikrein 6 and human kallikrein 10 protein levels are associated with younger age, pre-menopausal, status and tumours which are negative for oestrogen and progesterone receptors. No correlation was found between human kallikrein 6 and human kallikrein 10 levels and tumour size, grade, and nodal status. Survival analysis showed that neither human kallikrein 6 nor human kallikrein 10 are related to the rate of relapse-free and overall survival. In the analysis with respect to response to tamoxifen therapy, although human kallikrein 6 levels were not associated with tamoxifen responsiveness, higher levels of human kallikrein 10 were significantly associated with a poor response rate. This association remained significant in the multivariate analysis. Furthermore, higher human kallikrein 10 levels were significantly related with a short progression-free and post-relapse overall survival after start of tamoxifen treatment for advanced disease. Taken together, our results suggest that although human kallikrein 6 and human kallikrein 10 are not prognostic markers for breast cancer, human kallikrein 10 is an independent predictive marker for response of tamoxifen therapy. PMID- 12087469 TI - Heterogeneity of O6-alkylguanine DNA-alkyltransferase expression in human breast tumours. AB - An important determinant of cellular resistance to chemotherapeutic O6-alkylating agents, which comprise methylating and chloroethylating agents, is the ability of cells to repair alkylation damage at the O6-position of guanine in DNA. This is achieved by a specific DNA repair enzyme O6-alkylguanine DNA-alkyltransferase. In this study O6-alkylguanine DNA-alkyltransferase expression was measured in human breast tumours using both biochemical and immunohistochemical techniques. O6 alkylguanine DNA-alkyltransferase activity was then compared with known clinical prognostic indices to assess the potential role of O6-alkylguanine DNA alkyltransferase in predicting the behaviour of this common malignancy. The application of both biochemical and immunohistochemical techniques was feasible and practical. Most breast tumours expressed high levels of O6-alkylguanine DNA alkyltransferase. Immunohistochemical analysis showed marked variation in expression not only between individuals but also within individual tumours, and in the same patient, between metastases and between primary tumour and metastatic site. O6-alkylguanine DNA-alkyltransferase activity in tissue extracts significantly correlated not only with immunohistochemical staining intensity determined by subjective quantitation, but also with measures of protein levels using a computerised image analysis system including mean grey (P<0.001), percentage of cells positive for O6-alkylguanine DNA-alkyltransferase (P<0.001), and integrated optical density (P<0.001). O6-alkylguanine DNA-alkyltransferase expression did not correlate with any of the established clinical prognostic indicators for current treatment regimens. However, immunohistochemical offers a rapid and convenient method for assessing potential utility of O6-alkylating agents or O6-alkylguanine DNA-alkyltransferase inactivating agents in future studies of breast cancer treatment. PMID- 12087470 TI - Periodate-treated, non-anticoagulant heparin-carrying polystyrene (NAC-HCPS) affects angiogenesis and inhibits subcutaneous induced tumour growth and metastasis to the lung. AB - Periodate-treated, non-anticoagulant heparin-carrying polystyrene consists of about ten periodate-oxidized, alkaline-degraded low molecular weight-heparin chains linked to a polystyrene core and has a markedly lower anti-coagulant activity than heparin. In this study, we evaluated the effect of non anticoagulant heparin-carrying polystyrene on tumour growth and metastasis. Non anticoagulant heparin-carrying polystyrene has a higher activity to inhibit vascular endothelial growth factor-165-, fibroblast growth factor-2- or hepatocyte growth factor-induced human microvascular endothelial cell growth than heparin, ten periodate-oxidized-heparin and ten periodate-oxidized-low molecular weight-heparin, which is probably due to the heparin-clustering effect of non anticoagulant heparin-carrying polystyrene. Non-anticoagulant heparin-carrying polystyrene inhibited human microvascular endothelial cell, B16 melanoma and Lewis lung cancer cell adhesion to Matrigel-coated plates. Non-anticoagulant heparin-carrying polystyrene also showed strong inhibitory activities in the tubular formation of endothelial cells on Matrigel and B16-melanoma and Lewis lung cancer cell invasion in a Matrigel-coated chamber assay. In vivo studies showed that growth of subcutaneous induced tumours and lung metastasis of B16 melanoma and Lewis lung cancer cells were more effectively inhibited by non anticoagulant heparin-carrying polystyrene than ten periodate-oxidized-heparin and ten periodate-oxidized-low molecular weight-heparin. Furthermore, non anticoagulant heparin-carrying polystyrene markedly reduced the number of CD34 positive vessels in subcutaneous Lewis lung cancer tumours, indicating a strong inhibition of angiogenesis. These results suggest that non-anticoagulant heparin carrying polystyrene has an inhibitory activity on angiogenesis and tumour invasion and may be very useful in cancer therapy. PMID- 12087471 TI - Fibrinogen E fragment selectively disrupts the vasculature and inhibits the growth of tumours in a syngeneic murine model. AB - We recently demonstrated that a fragment of human fibrinogen, fibrinogen E fragment, inhibits the migration and differentiation of human endothelial cells in vitro. Here we show that it exerts similar effects on murine endothelial cells in vitro, and selectively disrupts tumour endothelium in vivo, causing widespread intravascular thrombosis and retarding the growth of CT26 tumours in a syngeneic murine model. PMID- 12087472 TI - DNA methylation and histone deacetylation associated with silencing DAP kinase gene expression in colorectal and gastric cancers. AB - Death-associated protein kinase is a positive regulator of programmed cell death induced by interferon gamma. To investigate the role of epigenetic inactivation of death-associated protein kinase in gastrointestinal cancer, we examined the methylation status of the 5' CpG island of the death-associated protein kinase gene. Methylation of the 5' CpG island was detected in 3 of 9 colorectal and 3 of 17 gastric cancer cell lines, while among primary tumours, it was detected in 4 of 28 (14%) colorectal and 4 of 27 (15%) gastric cancers. By contrast, methylation of the edge of the CpG island was detected in virtually every sample examined. Death-associated protein kinase expression was diminished in four cell lines that showed dense methylation of the 5' CpG island, and treatment with 5 aza-2'-deoxycitidine, a methyltransferase inhibitor, restored gene expression. Acetylation of histones H3 and H4 in the 5' region of the gene was assessed by chromatin immunoprecipitation and was found to correlate directly with gene expression and inversely with DNA methylation. Thus, aberrant DNA methylation and histone deacetylation of the 5' CpG island, but not the edge of the CpG island, appears to play a key role in silencing death-associated protein kinase expression in gastrointestinal malignancies. PMID- 12087473 TI - Serological identification and expression analysis of gastric cancer-associated genes. AB - Serological identification of tumour antigens by recombinant expression cloning has proved to be an effective strategy for the identification of cancer associated genes having a relevance to cancer aetiology and progression, and for defining possible targets for immunotherapeutic intervention. In the present study we applied this technique to identify immunogenic proteins for gastric cancer that resulted in isolation of 14 distinct serum-reactive antigens. In order to evaluate their role in tumourigenesis and assess the immunogenicity of the identified antigens, we characterised each cDNA clone by DNA sequence analysis, mRNA tissue distribution, comparison of mRNA levels in cancerous and adjacent non-cancerous tissues and the frequency of antibody responses in allogeneic patient and control sera. Previously unknown splice variants of TACC1 and an uncharacterised gene Ga50 were identified. The expression of a newly identified TACC1 isoform is restricted to brain and gastric cancer tissues. Comparison of mRNA levels by semi-quantitative RT-PCR revealed a relative overexpression of three genes in cancer tissues, including growth factor granulin and Tbdn-1--an orthologue of the mouse acetyltransferase gene which is associated with blood vessel development. An unusual DNA polymorphism--a three-nucleotide deletion was found in NUCB2 cDNA but its mRNA level was consistently decreased in gastric tumours compared with that in the adjacent non-cancerous tissues. This study has revealed several new gastric cancer candidate genes; additional studies are required to gain a deeper insight into their role in the tumorigenesis and their potential as therapeutic targets. PMID- 12087475 TI - Bread and butter public health: when things are just important. PMID- 12087474 TI - Zonal image analysis of tumour vascular perfusion, hypoxia, and necrosis. AB - A number of laboratories are utilising both hypoxia and perfusion markers to spatially quantify tumour oxygenation and vascular distributions, and scientists are increasingly turning to automated image analysis methods to quantify such interrelationships. In these studies, the presence of regions of necrosis in the immunohistochemical sections remains a potentially significant source of error. In the present work, frozen MCa-4 mammary tumour sections were used to obtain a series of corresponding image montages. Total vessels were identified using CD31 staining, perfused vessels by DiOC(7) staining, hypoxia by EF5/Cy3 uptake, and necrosis by haematoxylin and eosin staining. Our goal was to utilise image analysis techniques to spatially quantitate hypoxic marker binding as a function of distance from the nearest blood vessel. Several refinements to previous imaging methods are described: (1) hypoxia marker images are quantified in terms of their intensity levels, thus providing an analysis of the gradients in hypoxia with increasing distances from blood vessels, (2) zonal imaging masks are derived, which permit spatial sampling of images at precisely defined distances from blood vessels, as well as the omission of necrotic artifacts, (3) thresholding techniques are applied to omit holes in the tissue sections, and (4) distance mapping is utilised to define vascular spacing. PMID- 12087476 TI - Prevalence and related risk factors of areca quid chewing among junior high students in eastern Taiwan. AB - Areca quid chewing is a serious problem in eastern Taiwan. The prevalence and related risk factors of areca quid chewing among junior high students were studied. Group 1 consisted of 896 non-aboriginal students and group 2 of 565 aboriginal students. The students were asked to fill out a questionnaire anonymously. Among the non-aboriginal students, 2.2% were found as current areca quid chewers, 16.4% had tried but not become regular users, and 2.2% had used it regularly and quit, whereas 9.7% of aboriginal students were current chewers, and 32.7% had tried it but had not become regular users, and 7.4% had used it regularly and quit. The prevalence of areca chewing is significantly different between boys and girls in the non-aboriginal group. A logistic regression analysis indicated that a non-aboriginal student who is a male, grows areca at home, is a tobacco smoker, an alcohol drinker, and has family members and close friends as chewers is more likely to chew areca quid than others. Aboriginal students who are chewers are likely to grow areca at home, are tobacco smokers and alcohol drinkers, and have family members and close friends as chewers. Based on the findings of the study, the need for a health promotion program was identified. Areca quid is not only detrimental to people's health, but also to the environment in which people live. PMID- 12087477 TI - Evaluation of a self-rating screening test for areca quid abusers in Taiwan. AB - Areca quid chewing is a popular habit and areca is a well-known ethnopsychopharmalogic agent in southeast Asia. While the chewing habit is legal and also socially acceptable in many places of Taiwan, the public health problem of high oral cancer incidence has remained a priority on the health care list in our local health department. Helping areca quid chewers to reduce or even stop the habit will be paramount in the oral cancer prevention programme. Hence, in order to identify the appropriate strategy for stopping the chewing habit, it is important to distinguish whether an areca quid chewer has reached the level of substance abuse.In accordance with the Diagnostic and Statistical manual of Mental disorder (fourth edition, DSM-IV), we developed a specific self-report questionnaire modified from the famous SCAN system, DSM-IV and ICD-10. The initial screening test for areca quid abusers had 52 questions. Its components included the onset age and frequency, subjective craving and feeling, social problems, physical problems, oral symptoms, psychological and abstinence-related problems, the motivation and capacity to abstain, and demographic data. The answers were divided into 'Yes' or 'No'. One hundred and twenty-five areca quid users (53 men, 72 women) were recruited. The abusers tended to have older age, less education, and higher daily consumption of areca quid. There were no differences on motivation to quit chewing (abstinence) between abusers and non abusers. There were no statistical differences on tobacco-smoking and alcohol drinking behavior. Based on the statistical analysis of receiver operation characteristic (ROC) curves, 11 questions were chosen for the Self-report Screening Test for Areca quid Abuser (SSTAA). An areca quid chewer's answers with a score of 4 or more in these 11 questions would be considered an areca quid abuser. The modified process of SSTAA is performed for the evaluation of the native culture-related substance user. At this current stage, the SSTAA developed from the study is the first instrument for screening areca quid abuser. PMID- 12087478 TI - Mapping the public health workforce I: a tool for classifying the public health workforce. AB - We aimed to develop a tool to identify members of the public health workforce and classify them using categories developed for the Chief Medical Officer's project to strengthen the public health function. The tool was developed to gain a picture of London's public health workforce, and needed to be reliable and easy to use in many settings inside and outside the health service. We needed to be able to classify posts from brief information without interrogation of postholders, so that the entire workforces of large organisations could be classified from information provided by only a few key informants. Key questions and decision rules were defined by presenting interviewees in public health with brief information on nine jobs and discussing with them the process by which they determined whether each post was in the public health workforce, and if so, in which category. The questions and decision rules were refined into a classification tool which was presented as a flow diagram and a questionnaire. Application of the tool revealed that it was understood by key informants and resulted in classifications which were accepted by the researchers. The tool has now been applied extensively in London and yielded useful results. Many other applications in public health workforce planning and development are anticipated. PMID- 12087479 TI - Perceptions of partnership. A documentary analysis of Health Improvement Programmes. AB - Health Improvement Programmes (HImPs) are at the heart of the UK government's partnership agenda for the National Health Service (NHS). This paper assesses the nature of HImP partnerships in England by analysing 50/99 first-round HImP strategies (randomly selected). The documentary analysis quantifies the structures and mechanisms of partnership, the degree of inter-sectoral participation and the extent of voluntary sector involvement.Three-quarters of responding health authorities (37/50) appear to have set up formal partnership structures to produce the HImP, or are planning to do so. After health authorities, local authorities (47/50) appear to be most involved in contributing to the HImP, particularly social services departments. Within the NHS 'family', acute and community trusts (43/50) appear to be the most involved, with Primary Care Groups (PCGs) contributing less (39/40). Community Health Councils (CHCs) appear to be similarly involved (40/50). The voluntary sector appear to be involved in all but four HImPs, mainly through umbrella organisations represented on strategic partnership boards (34/50). User and carer and community groups appear to participate far less. Lack of endorsement of HImPs by partner organisations, poor delineation of responsibilities and absence of transparency in resource allocation suggest that ownership of, and commitment to HImPs may be weak. HImPs appear to have focused on creating structures rather than developing aspects of partnership process. If levels of inter-sectoral involvement and voluntary sector participation are to be maintained or increased in future, Primary Care Trusts (PCTs) will need to develop a strategic approach to partnership. PMID- 12087480 TI - Nurse-led welfare benefits screening in a General Practice located in a deprived area. AB - OBJECTIVE: To evaluate a nurse-led attendance allowance screening service in General Practice. DESIGN: Intervention study. SETTING: One General Practice located in two sites in deprived areas in the East-End of Glasgow (Carstairs Deprivation Index-6 and 4). PARTICIPANTS: Participants aged >64 y who in the nurses clinical judgment appeared to be physically or mentally frail were opportunistically recruited over a 12-week period by community nurses (health visitors, district nurses and practice nurses). A Welfare Rights Officer (WRO) contacted all potential underclaimers by telephone and offered a home visit in order to assess for unclaimed benefits. MAIN OUTCOME MEASURE: Total unclaimed attendance allowance and linked benefits. RESULTS: Thirty-seven of the original 86 participants plus four relatives were found not to be claiming the benefit payments that they were entitled to. Referral to the Department of Social Security (DSS) revealed unclaimed benefits to a total of: pound sterling112 893.00of this pound sterling95 306.00 is on a recurrent annualized basis and pound sterling17 587.00 as lump sums. CONCLUSIONS: A community nurse-led attendance allowance screening service combined with a home visit by a WRO was an efficient and highly effective model for maximising the income of the frail elderly. This model could contribute to reducing the increasing number of pensioners living below the poverty line. PMID- 12087481 TI - An interim report of health needs assessment of asylum seekers in Sunderland and North Tyneside. AB - A health needs assessment questionnaire was developed and administered to 397 asylum seekers, 291 males and 106 females, in Sunderland and North Tyneside in the North East of England. The data from the questionnaires were analysed using SPSS Version 10. Individuals retained a copy of their own questionnaire to be used as a hand held record. There was much heterogeneity of country of origin, culture, religion and previous employment and language backgrounds within the sample. A range of health needs was identified which has implications for healthcare provision. Vaccination rates were low as was screening for tuberculosis and cervical cancer. Many asylum seekers identified symptoms related to mental health and requested help in this area. Access to dental treatment was an area of high priority for many respondents. PMID- 12087482 TI - Leisure time activity as a determinant of survival: a 26-year follow-up of a Swedish cohort. AB - An individual's leisure time (pastime) engagements are in a way important for society. Irrespective of whether leisure time activities are causal determinants of health or health is a prerequisite for taking full part in society, the interaction is a challenge for Public Health. The first question is whether the sum of their influence results in coherence between enjoying good health and having leisure time activity. The aim of this study was to estimate their covariance.A random sample was drawn from the adult population of Stockholm County, Sweden. The residents were mailed a questionnaire regarding their social circumstances, their health complaints, the social repercussions of the complaints, and if they had any leisure time activity. Altogether, 7252 (about 93%) individuals responding to the questionnaire constituted our cohort. The individuals aged 18-65 y in November 1969 were followed up to the age of 65 y, or to 1996 as to mortality. The main outcome measure was mortality irrespective of cause of death. More than two-thirds of the respondents (71%) reported that they had some leisure time activity. Leisure time activity was a determinant of survival in the statistical sense. The risk ratio was 0.77 and its 95% confidence interval was 0.68-0.87 for those reporting leisure time activity when age, sex, baseline health and baseline alcohol consumption was discounted. The conclusion was that having leisure time activity, unspecified, covaried with health. Further studies should specify the pastime activities. PMID- 12087483 TI - Physical disability in The Netherlands: prevalence, risk groups and time trends. AB - Physical disability represents an important health indicator of western populations. In this paper the prevalence of physical disabilities in The Netherlands is presented for four domains of disability-visual, hearing, mobility and activities of daily living (ADL) disability-with a focus on risk groups and time trends.Cross-sectional national health survey data (NetHIS) of 9 y, 1990 1998, presenting data on 62 352 persons of 16 y or over were used. All data were self-reported. About one-eighth of the research population had a physical disability, ie had at least major difficulty with one or more functions such as walking, seeing, hearing and washing. This figure increased from 1.7% in the age group of 16-24 y to 44.1% in the age group of 75 y or older. Risk groups were women, those living alone, those who were divorced or widowed and those with a low educational level. In the period 1990-1998, the prevalence did not change with the exception of the prevalence of mobility disability which dropped slightly with 0.2 percentage points per year due to decreasing prevalences among men. One conclusion is that the prevalence of disability is high and stable, and expected to increase in the future due to the ageing of the population. PMID- 12087484 TI - Integrating prevention of acute respiratory infections with micro-credit programme: experience of BRAC, Bangladesh. AB - The contribution of acute respiratory infection control project within the framework of micro-credit-based development intervention in promoting maternal knowledge of ARIs in children was assessed. Data came from a cross-sectional survey of 2814 mothers of under 5-y-old children residing in 200 randomly selected villages in five districts in Bangladesh. Findings revealed that the ARI control project had significant positive effects in raising knowledge of clinical signs and preventive measures. When ARI control project activities were integrated with the credit-based development initiative, maternal knowledge improved even further. The study concludes that the micro-credit programme can be a catalytic agent in raising health knowledge among poor women in developing countries. PMID- 12087485 TI - Beliefs and behaviours of Iranian teachers toward early detection of breast cancer and breast self-examination. AB - A descriptive study was carried out to explore the beliefs and behaviours of Iranian teachers toward breast cancer and breast self-examination (BSE). A 10% random sample of 8000 teachers working in four central regions of Tehran was selected. Data on beliefs and behaviours regarding breast cancer and BSE were collected by means of a structured questionnaire. In all, 578 women participated in the study. The mean age of the respondents was 37.3 y (s.d.=8.8), and most were married (83%). The majority of the respondents believed that 'Like every other woman I am at risk of breast cancer' (67%) and that 'Family history of breast cancer is a risk factor for breast cancer' (59%). However, only 6% of them reported that they are performing BSE on a regular basis. The most common reasons for not doing BSE and not going for clinical examination were lack of knowledge and the belief that it was not necessary (34 and 36%, respectively). Doing BSE was more frequent in married (P<0.00001), and older (P<0.00001) women. Except for those with a positive family history of breast cancer, there was no association between breast cancer beliefs and almost all other variables studied. The findings suggest that although the majority of Iranian teachers seem to be quite knowledgeable about breast cancer, they need more education on breast cancer and BSE. Since teachers' beliefs and behaviours may have an impact on young females, planning training courses for this group of women is essential. PMID- 12087487 TI - Endoplasmic reticulum storage diseases. AB - The endoplasmic reticulum represents the cell's quality control site for accurate folding of secretory and membrane proteins. Quality control is achieved through the association of ER chaperones with unfolded or misfolded polypeptide chains. In the ER stress response, upregulation of chaperones occurs as a consequence of misfolded proteins accumulating in the ER lumen; if these proteins fail to assume their native structure, they are retained in the ER and targeted for degradation by the proteasome. ER storage diseases (ERSDs) are a group of genetically based disorders in which mutant proteins fail to pass the ER quality control. Because all eukaryotic cells contain the ER, the clinical phenotype of ERSDs is very heterogeneous. Disease may result from the mere lack of the mutant protein in question and/or may be caused indirectly by toxic effects of the misfolded protein or aggregates thereof on the cell. Additionally, the cell's reaction to the ER stress may include signaling pathways which are ultimately detrimental. Experimentally, ERSDs serve as models to study the cellular reactions to a variety of perturbations. In particular, understanding the links between ER stress and cell degeneration may give valuable insights into the pathogenesis of other diseases where the accumulation of indigestible toxic material leads to cell injury. PMID- 12087488 TI - Epidemiology of methicillin-resistant Staphylococcus aureus: results of a nation wide survey in Switzerland. AB - OBJECTIVE: To assess the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) in Switzerland. MATERIAL AND METHODS: One-year national survey of all MRSA cases detected in a large sample of Swiss healthcare institutions (HCI). Analysis of epidemiological and molecular typing data (PFGE) of MRSA strains. RESULTS: During 1997, 385 cases of MRSA were recorded in the 5 university hospitals, in 33 acute care community hospitals, and 14 rehabilitation or long term care institutions. Half of the cases were found at the University of Geneva Hospitals where MRSA was already known to be endemic (41.1 cases/10,000 admissions). The remaining cases (200) were distributed throughout Switzerland. The highest rates (>100 cases/10,000 admissions) were reported from non-acute care institutions. Rates ranged from 3.3 to 41.1 cases/10,000 admissions for university hospitals (mean 15.5); 0.67 to 90.4 for community hospitals (mean 4.8), and 28.2 to 315 for non-acute care institutions reporting MRSA (mean 85.7). Forty percent of MRSA patients were infected, while 60% were only colonised. The leading infection sites were skin and soft tissue (21%), surgical site (15%), and the urinary tract (26%). Whereas in Eastern Swiss HCI most MRSA cases occurred in acute care hospitals (n = 47, 98%), rehabilitation and long-term care institutions accounted for an important number of the identified cases (n = 107, 38%) in Western Switzerland. CONCLUSION: Low rates of MRSA were still observed in Swiss HCI, despite one outlying acute care centre with endemic MRSA and some nonacute care institutions with epidemic MRSA. Rehabilitation and long-term care institutions contributed to a substantial proportion of cases in Western Switzerland and may constitute a significant reservoir. Overall, a national approach to surveillance and control of MRSA is mandatory in order to preserve a still favourable situation, and to decrease the risk of epidemic MRSA dissemination. PMID- 12087490 TI - Experiences with interferon-beta-1b treatment in MS after three year follow-up. PMID- 12087489 TI - Reactive macrophage activation syndrome: a simple screening strategy and its potential in early treatment initiation. AB - QUESTIONS UNDER STUDY: starting treatment of reactive macrophage activation syndromes as early as possible (rMAS, haemophagocytic lymphohistiocytosis), e.g., with intravenous immunoglobulins (IVIG), seems to be essential for optimal outcome. However, there is no diagnostic gold standard which reliably indicates need for early treatment. We used a simple screening strategy consisting of serum ferritin measurements and/or morphological assessment of haemophagocytosis and compared the studied patient population with published series. METHODS: Retrospective analysis of clinical and laboratory data of 57 patients experiencing 60 episodes of rMAS. RESULTS: Screening by serum ferritin measurements and/or morphological assessment of haemophagocytosis of patients presenting with a systemic inflammatory response syndrome (SIRS) indicates that rMAS might be considerably more frequent than stated in the literature. Serum ferritin exceeded >10,000 microg/L in 91% rMAS episodes. Although the patient population studied was otherwise similar in most aspects to the published rMAS series, the fact that 40% of patients fulfilled the criteria for Still's disease (SD) as the disorder underlying rMAS is remarkable and questions the distinct nature of the two diseases. IVIG responders and non-responders did not differ regarding their initial characteristics with exception to the timepoint of IVIG administration, confirming the importance of early treatment initiation. Malignancy-associated rMAS however, has a poor prognosis and seems to be refractory to manipulation with IVIG in most instances, even when responding initially. CONCLUSIONS: rMAS has to be considered in patients with a SIRS- or SD like clinical presentation. Hyperferritinaemia >or=10,000 microg/l seems to be a good marker for defining patients with or at risk for developing rMAS and should be completed with a morphological assessment of haemophagocytosis. The perception of acute SD and rMAS as two distinct entities has to be questioned at least in a subgroup of patients. PMID- 12087493 TI - When did leptin become a reproductive hormone? AB - Almost 50 years ago the obese mouse model was identified, and parabiosis studies were able to demonstrate that some humoral factor was involved in adiposity, so that the genetics and endocrine nature of this process have been apparent for many years. With the discovery of leptin just a few years ago, early studies validated the role of this protein product of the obese gene. Early studies in the obese mouse model ( ob/ob mouse) demonstrated that the genetic basis was truly a deficiency in leptin. Coincidentally, the relationship to fertility was also associated with leptin. These early studies were also able to demonstrate a relationship to puberty and the time of pubertal development. Very quickly, the recognition that the placenta was a source of leptin and that leptin levels were elevated in pregnancy in a number of species also broadened our appreciation of the relationship to reproductive functions. These many rapidly elucidated relationships to leptin were reported soon after the identification and availability of leptin as a research reagent and have firmly put leptin into the area of reproductive physiology in addition to establishing roles in metabolism, satiety, and energy metabolism. Subsequent studies have expanded all of these situations. Species beyond the rodent model, including the human, have now introduced these physiologic studies into the clinical arena and the role of leptin in fertility, puberty, pregnancy, and genetics. In this issue, all of these topics are reviewed to bring the reader up to date with leptin and its role in reproductive function, many of which overlap with the control of obesity. PMID- 12087494 TI - Leptin and pubertal development. AB - Sexual development after birth in rodents, nonhuman primates, and humans is driven by the gonadotropin-releasing hormone (GnRH) pulse generator. During the neonatal period in primates, pulsatile GnRH discharge from the medial basal hypothalamus drives an active period of pituitary gonadotropin and gonadal hormone secretion. During the transition from the neonatal to the juvenile period, however, the activity of the GnRH pulse generator is restrained or arrested and gonadotropin and gonadal hormone secretion enters a quiescent period that continues until the onset of puberty. As puberty approaches the GnRH pulse generator is reactivated, resulting in enhanced gonadotropin secretion, accelerated growth, maturation of the gonads, and the achievement of sexual competence. Rodents do not appear to exhibit a developmental phase analogous to the quiescent juvenile period in primates when the GnRH pulse generator is held in check. Instead, progressive maturational changes in the pattern of GnRH pulsatility appear to drive sexual development in rodents. The role that leptin plays in sexual development has not been fully defined, but the balance of current evidence appears to support the idea that, in both rodents and primates, leptin plays a permissive rather than a causal role in timing this process. When body energy reserves rise above a critical level, blood leptin increases to a threshold concentration signaling to the central nervous system that the body can support sexual function. Puberty can apparently occur over a wide range of concentrations above this critical leptin threshold. Leptin does not appear to act as a trigger to time the initiation of puberty but, instead, once leptin reaches this threshold pubertal development may proceed if, and only if, other critical control mechanisms are operational. PMID- 12087496 TI - Leptin: roles and regulation in primate pregnancy. AB - Leptin, a hormone produced by adipose tissue and the placenta, is enhanced in the maternal circulation throughout pregnancy in both the human and the baboon ( Papio sp.), a proven nonhuman primate model for the study of human pregnancy. The presence of both leptin and its receptor in the fetus implies a role for the polypeptide as a regulator of in utero development, although localization in the placental trophoblast may relate to autocrine and/or paracrine regulatory functions in this important endocrine tissue. Although regulatory mechanisms remain incompletely defined, it has been suggested that cross talk occurs between the fetus, placenta, and maternal adipose stores. Placental estrogen, which is present in increasing concentrations with advancing gestation, is suggested to influence leptin synthesis in a tissue- and cell type-specific fashion. In this capacity, cellular hypoxia, diabetes, and preeclampsia are conditions that appear to be intimately linked to leptin dynamics. A better understanding of regulatory mechanisms will have direct clinical significance, as leptin has been proposed to impact on those causes of human perinatal morbidity and mortality that are associated with anomalies of fetal maturity and development, general conceptus growth, trophoblast endocrinology, and placental sufficiency. PMID- 12087495 TI - Leptin and ovarian folliculogenesis: implications for ovulation induction and ART outcomes. AB - Leptin participates in regulation of ovarian folliculogenesis indirectly via control of luteinizing hormone and follicle-stimulating hormone secretion. More recent evidence suggests that leptin also has direct regulatory actions on the developing follicle. The presence of leptin receptors on follicular cells, including oocytes, and early preimplantation embryos suggests that leptin may play a direct physiologic role in follicular maturation, oocyte development, and early cleavage. Because circulating leptin levels are directly related to body adiposity, elevated leptin concentrations associated with obesity may partly explain the negative impact of obesity on fertility. The influence of leptin on follicular development and oocyte maturation has important implications for ovulation induction and assisted reproductive technologies. Moreover, polycystic ovarian syndrome may be associated with altered leptin phsyiology. PMID- 12087497 TI - Leptin in relation to growth and developmental processes in the fetus. AB - Leptin, a 16-kilodalton protein secreted by the adipose tissue in proportion to the amount of energy stored in adipose tissue, conveys to the hypothalamus information on energy homeostasis and regulates reproductive function. In addition, there is accumulating evidence that leptin produced by placental or fetal tissues acts through specific leptin receptors to regulate fetal growth and development. Although leptin levels are correlated with insulin and insulin-like growth factor (IGF)-1 levels, observational studies in humans indicate that its effects on fetal growth are independent of these axes and of adiposity. The extent to which leptin per se mediates the fetal growth and developmental abnormalities associated with disease states such as diabetes, hypoxia, or preeclampsia remains to be fully clarified by future studies in humans. It is hoped that clarification of these mechanisms may provide novel therapeutic approaches. PMID- 12087498 TI - Leptin and preeclampsia. AB - Preeclampsia, a common complications of pregnancy, is associated with an increase in the concentration of leptin in the maternal blood, which precedes the clinical onset of the disease. This review addresses the potential sources of leptin and considers the possible consequences, although at present these are entirely conjectural. The placenta is likely to contribute to the rise in leptin, and placental hypoxia and inflammatory mediators may be important stimuli. The possible protective and damaging sequelae of an increase in the maternal leptin concentrations may range from beneficial stimulation of fetal growth to an increase in blood pressure through stimulation of sympathetic activity. Further research is needed to determine if the rise in leptin plays a role in preeclampsia or whether it is a secondary and unrelated bystander. PMID- 12087499 TI - Leptin and the onset of puberty: insights from rodent and human genetics. AB - Deficiency of the adipocyte-derived hormone leptin in ob/ob mice results in severe, early-onset obesity and infertility. Administration of leptin results in complete reversal of the phenotype, suggesting that leptin is needed for the development of puberty in rodents. In humans, mutations in the genes encoding leptin and the leptin receptor result in obesity syndromes and hypogonadotropic hypogonadism. I have shown that administration of recombinant human leptin results in the onset of puberty at an appropriate developmental age in human congenital leptin deficiency. This work suggests that leptin is a metabolic gate for the onset of puberty in humans. Leptin's actions may be mediated by central pathways and by direct action on peripheral organs. PMID- 12087500 TI - Leptin and reproductive function in males. AB - Leptin is a circulating protein produced by adipocytes that has been implicated in control of body weight through appetite regulation and control of reproduction, most likely through an effect on the central nervous system. From studies in mice, it is clear that the genetic background of the animal on which the mutation in the leptin gene is placed can influence how that mutation is expressed. Although the effects of leptin have been more thoroughly documented in nonprimate species than in primates, a few human families with genetic mutations of the gene for leptin or the leptin receptor show obesity and impaired fertility. There is conflicting evidence regarding the effects of leptin in male primates, but it appears that the metabolic and reproductive effects of low leptin levels caused by reduced energy intake in adult animals can be more readily alleviated by administration of energy substrates than by administration of leptin. PMID- 12087501 TI - Ventricular failure as a cause of unsuccessful endoscopic third ventriculostomy. AB - INTRODUCTION: In spite of extensive studies on cerebrospinal fluid (CSF) dynamics, the mechanism of its circulation is still obscure. The aim of this study is to evaluate the effect of an insult to the brain tissue on the success or failure of an endoscopic procedure. METHODS: During the period from May 1995 to December 1998 we studied 21 children, ages from 2 to 48 months (mean 15 months) with non-communicating hydrocephalus. The intracranial pressure was measured at the time of endoscopic surgery. The follow-up period was from 2 - 5 years. They were divided into 2 groups. Group I were 8 patients with no history of brain insult. Group II (13 cases) had a previous brain insult (infective and/or vascular). Endoscopic third ventriculostomy (ETV) was done in 17 cases; 5 in Group I and 12 in Group II. Fenestration of cyst/s was done in all of Group I and in 9 cases of Group II. Failure was considered whenever shunt implantation was required. RESULTS: The mean intracranial pressure in Group I was 13.0 mmHg as compared to 9.7 mmHg in Group II (p = 0.015). The failure rate of the endoscopic procedure/s was 25 % and 54 % in Groups I and II, respectively. DISCUSSION: The success of ETV and/or fenestration of cyst/s depends on a sufficient pressure gradient. The pressure gradient is generated by a normal or compensated ventricular function. CONCLUSION: Brain tissue damage can result in ventricular failure. The quality of ventricular wall function has a significant role on the success of ETV. PMID- 12087502 TI - Relevance of magnetic resonance imaging for ventricular endoscopy. AB - OBJECTIVE: Endoscopic procedures may represent an alternative to microsurgery or shunt operations in intra- or paraventricular pathologies. However, the operative results mainly depend on the selection of suitable patients. In the present study of 30 patients with occlusive hydrocephalus, the relevance of differentiated magnetic resonance (MR) imaging for evaluation of the pathophysiological and anatomic prerequisites was investigated. PATIENTS AND METHODS: Apart from conventional axial T(2) -weighted turbo spin echo (TSE) sequences (TR/TE 2126/100 ms) and T(1) -weighted spin echo (SE) sequences (TR/TE 450/15 ms) with and without the administration of Gd-DTPA, the MR protocol included sagittal and coronal thin-sliced T(2) -weighted TSE sequences, cardiac-triggered (TR/TE > 2700/120 ms, slice thickness 3.0 mm) or in volume technique (TR/TE 4000/180 ms, slice thickness 1.4 mm). For the visualization of CSF flow, a sagittal and axial phase-contrast-multi-heart-phase (PCMHP) measurement with 16 phases per cardiac cycle and flow sensitivity in the cranio-caudal direction (TR/TE 18/9.5 ms and 14/9.5 ms) was performed. RESULTS: The combination of imaging and flow sequences provided most information about the extent and the etiology of hydrocephalus. An obstruction of CSF pathways could be detected by the absence of flow voids in T(2) -weighted TSE sequences or of flow signals in PCMHP sequences. The size of the foramen of Monro and of the third ventricle could be evaluated in thin-sliced T(2) -weighted TSE sequences, for the foramen of Monro in coronal and sagittal and for the third ventricle in coronal and axial slice direction, respectively. The basal structures of the third ventricle, including its floor, the infundibular recess, the mamillary bodies, and the basilar artery, could be best identified in sagittal thin-sliced T(2) -weighted TSE scans, cardiac-triggered or in volume technique. CONCLUSION: In occlusive hydrocephalus, a differentiated MR protocol with thin-sliced T(2) -weighted TSE sequences and phase-contrast sequences allows a very precise estimation of the pathophysiological and the anatomic prerequisites for endoscopic procedures. Furthermore, MR imaging can be used for the follow-up documentation of ventriculo-cisternostomies, -cytostomies, or openings of other membranes. PMID- 12087503 TI - Uniportal endoscopic surgery of carpal tunnel syndrome: technique and clinical results. AB - The endoscopic carpal tunnel release is a new minimally invasive method which has been used to decompress the median nerve in the carpal tunnel for the past decade. Advantages of this method should be a decreased postoperative morbidity and earlier return to work. Preoperative complaints, postoperative results and complications of the therapy for a total of 60 patients are presented. All endoscopic releases were performed using the Agee uniportal technique. The overall success rate in our study was 56/60 (93.3 %). 47/54 (87.0 %) patients were completely free of pain after endoscopic surgery. An improvement in preoperative pain was noted in 4/54 (7.4 %) patients. Hypesthesia and dysesthesia disappeared totally in 39/46 (84.8 %) patients. An improvement of the sensible disturbances was observed in 4/46 (8.7 %) cases. 10/13 (76.9 %) preoperative pareses recovered completely, 3/13 (23.1 %) remained unchanged. The complication rate in total in our series was 4/60 (6.7 %), thereof 3 cases of post-operative infection (5 %) and one serious median nerve injury (1.7 %). The mean time for return to work was 29 days. Summing up, it may be said that monoportal endoscopic carpal tunnel release appears to be an effective and safe minimally invasive method for the treatment of carpal tunnel syndrome. PMID- 12087504 TI - Endoscopic reverse third ventriculostomy via the cisterna magna: anatomical study and proposal of a novel procedure. AB - Basilar artery injury has been known as a potential lethal complication of endoscopic third ventriculostomy. In order to avoid this complication, endoscopic reverse third ventriculostomy via a trans-cisterna-magna route was studied. A cadaveric study was performed for navigation of a flexible endoscope through the cisterna magna. Three fresh, unfixed cadavers were used for this endoscopic navigation. In the prone position, a small vertical paramedian skin incision is made at the mid-portion of the posterior neck. An 11-mm threaded plastic tube is inserted towards the posterior arch of the atlas. After a partial hemilaminectomy of the atlas, a flexible endoscope is introduced into the cisterna magna and is navigated cephalad along the vertebrobasilar artery to the inferior aspect of the floor of the third ventricle. Through the working channel of a fiberscope, third ventriculostomy is performed in a reverse direction. Additional detailed anatomy was studied in fixed cadaveric head specimens with a rigid rod-lens endoscope for anatomic orientation. A novel technique of a trans-cisterna-magna reverse third ventriculostomy was studied in cadaveric specimens. This technique may avoid basilar artery injury which occurs occasionally during conventional third ventriculostomy. PMID- 12087505 TI - Computer-assisted resection of supra-tentorial cavernous malformation. AB - INTRODUCTION: Resection of cerebral tumors or vascular lesions requires a precise localization to minimize the skin, bone and cerebral approach. The image-guided surgery is currently considered to be of undisputed value in microneurosurgical technique. METHODS: Between 1998 and 2000, 13 patients were operated in our service for resection of a cavernous malformation deeply located using the MRI assisted image guidance (Sofamor-Danek Neuronavigation Cranial 3 System). RESULTS: The computer-calculated registration accuracy ranged between 0.8 and 2.0 mm (median 1.1 mm). The exact location of the cavernous malformation was possible in all the cases. Total resection of the lesion was always achieved. Operative mortality and transient morbidity were 0 % and 16 %, respectively. DISCUSSION: The image-guided technique offers a better help than the previously used methods (preoperative localization with CT scan or stereotactic implantation of guiding catheters) to resect intracranial lesions, especially if the lesion is deeply situated in the brain or in an eloquent area. Preoperative MRI-based 3D models, performed using special skin markers, and surgical computer-assisted neuronavigation allow us to find and to resect small and deep lesions with minimal mortality and low morbidity rate. PMID- 12087506 TI - High frequency stimulation of the basal ganglia for the treatment of movement disorders: current status and clinical results. AB - High frequency stimulation of the basal ganglia has gained much interest during the last years. Based on the reevaluation of the results of functional neurosurgery for movement disorders from Leksell's group, pallidotomy as lesional procedure was the first functional operation that underwent a renaissance for the treatment of movement disorders. The work by Benabid and Siegfried who carried out thalamic high frequency stimulation to suppress tremor, the knowledge about deep brain stimulation (DBS) for the treatment of chronic pain as well as better understanding in basal ganglia physiology and the development of reliable stimulation hardware led to an increasing number of centers worldwide who currently apply high frequency DBS for different movement disorders. In the present review the current status of DBS for movement disorders is presented and the results with high frequency stimulation targeted at different brain areas are summarized. PMID- 12087507 TI - Continuous neural monitoring in lumbar spine surgery: experience with 101 patients. AB - The first report about lumbar spinal surgery under epidural anesthesia has been published in 1958. The aim of this study was assess to epidural anesthesia as a regional operative anesthesia in lower lumbar spine surgery and to use the opportunity of verbal interaction with the patient who is able to move her/his lower extremities voluntarily during surgery. We called this simultaneous clinical assessment process as continuous neural monitoring. Established pathologies in the patients were as follows; lumbar herniated intervertebral disc disease (n = 95), lumbar spondylolisthesis (n = 4), lumbar lamina fracture (n = 1) and lumbar dumbbell Ewing's sarcoma (n = 1). Preoperative findings, operative findings, operative complications, postoperative complications and short-term results were assessed by the surgeon-anesthesiologist team. The patients, 99 out of 101, were followed-up for more than 15 months (mean 21.2 months). Fair and failure results were assessed in 7.0 % of the patients. Even though epidural anesthesia is almost equal to general anesthesia in terms of anesthetic complications, surgical complications and surgical results in uncomplicated lumbar spinal surgery cases, it has the advantage of being able to simultaneously evaluate motor results of surgical maneuvers and/or manipulations on the neural structures. This advantage, which is not available at the same accuracy with other neurophysiologic monitoring modalities, would be valuable in complex lumbar spine surgery cases. PMID- 12087508 TI - The combined subtemporal-suboccipital approach: a modified surgical access to the clivus and petrous apex. AB - OBJECTIVE: The surgical access to the clivus and the petrous apex is still a challenge. A combined approach is best fitted to lesions located in the middle and posterior cranial fossa. The approach described is centered on the petrous bone and requires an extensive bone resection; nevertheless, no osteoplastic bone flap is necessary. METHODS: In contrast to approaches described before, the petrous bone is drilled away anterior to the sigmoid sinus more extensively, the sinus is unroofed. For exposure of the middle cranial fossa the petrous bone should be resected down to the roof of the external meatus, the total extent of the craniotomy is significantly smaller. RESULTS AND CONCLUSION: The surgical access as described above provides a wide operative field under preservation of important intracranial structures. This modified approach minimizes the cerebellar and temporal lobe retraction. The neural and vascular structures can be preserved under direct vision to the tumor. The blood supply is interrupted at the beginning of the operation. PMID- 12087509 TI - Endoscopic fenestration of symptomatic septum pellucidum cysts: three case reports with discussion on the approaches and technique. AB - OBJECTIVES: We describe three patients with symptomatic septum pellucidum cysts treated by endoscopic fenestration and discuss the different endoscopic approaches to these cysts. CLINICAL PRESENTATION: The patients are an 8-year-old boy, a 7-year-old boy and a 21-year-old woman; this last also had a right frontal cavernous angioma and a pituitary microadenoma. All patients presented with headache, associated with vomiting in two and behavioral changes in one. In all cases magnetic resonance showed a septum pellucidum cyst. INTERVENTION: Two patients were operated upon by posterior approach through a right occipital burr hole and underwent fenestration from the right occipital horn to the cyst, with a second fenestration from the cyst to the left lateral ventricle in one. Another patient underwent microsurgical removal of a right frontal cavernoma and endoscopic cyst fenestration with both lateral ventricles through a right frontal craniotomy. Postoperatively, headache and vomiting resolved in all cases and behaviour changes improved in one. CONCLUSIONS: Endoscopic fenestration is the treatment of choice for septum pellucidum cysts, where it results in immediate relief of the mass effect of the cyst and in the remission of the associated symptoms. We suggest a posterior approach through a right occipital burr hole. It allows one to easily cannulate the occipital horn, which is usually larger than the frontal one, thus avoiding the risk of damaging the vascular and neural structures surrounding the foramen of Monro. Besides, the endoscopic trajectory is in our opinion more direct. The two-window technique, with fenestration of the cyst into both lateral ventricles, improves the chances of long-term patency. PMID- 12087510 TI - Simultaneous treatment of a pituitary adenoma and an internal carotid artery aneurysm through a supraorbital keyhole approach. AB - We describe a case in which an incidental left carotid aneurysm was found during the preoperative evaluation of a pituitary adenoma. After magnetic resonance imaging (MRI) and angiography delineated their characteristics, we chose to treat both lesions simultaneously through a left supraorbital keyhole approach. The aneurysm clipping was followed by intracapsular resection of the tumor that was identified later as a non-secretory adenoma. The postoperative course was uneventful. This case shows that in carefully selected patients it is possible to treat successfully concurrent conditions through a minimally invasive approach, with the well known advantages of low morbidity and a short hospital stay. PMID- 12087511 TI - Combined surgical and intraoperative endovascular approach for a giant internal carotid artery aneurysm in the high cervical region. AB - For the obliteration of a large aneurysm located at the cranial base or high cervical region, several therapeutic strategies including a parent vessel ligation, and endovascular occlusion have been reported, because it is difficult to access the aneurysm itself. We used a combined surgical and endovascular approach for the treatment of a large internal carotid artery aneurysm in the high cervical region. In the present case, we performed superficial temporal artery to middle cerebral artery bypass, then obliterated the aneurysm with distal coil embolization and proximal ligation in one session, using portable digital subtraction angiography. The combined endovascular and surgical approach involves less invasive surgery for complex cerebrovascular lesions. PMID- 12087512 TI - Endoscopic removal of third ventricular tumors: a technical note. AB - OBJECT: An endoscopic surgical technique utilizing a rollable vinyl tube as a surgical corridor is described for removal of third ventricular tumors. SURGICAL TECHNIQUE: Transcortical transventricular access is made via a burr hole placed at the point which is one inch lateral from the midline at the coronal suture area. A one-cm-diameter vinyl tube is slit longitudinally, rolled like a cigarette, and tied at its ventricular end with a release tie. With image-guided stereotactic assistance, this rolled vinyl tube is inserted into the lateral ventricle near the foramen of Monro. When the release tie is untied, the vinyl tube expands to its original 10-mm-diameter tube by recoil assisted with a balloon dilatation technique. Through this tube, a rod-lens endoscope is placed to visualize the surgical target and is mounted to an endoscope holder. Surgical instruments are inserted next to the endoscope for surgical dissection. Compared to endoscopy through fixed working-channel devices, this technique allows increased flexibility for the surgeon when maneuvering surgical instruments for delicate dissection and tumor removal. Two patients with colloid cysts and one patient with an epidermoid tumor are reported as demonstrative cases. CONCLUSION: An endoscopic technique utilizing a soft vinyl tube which can be rolled into a small diameter and then unrolled to its original size by its own recoil when a release tie is removed and by balloon dilatation, is reported for removal of third ventricular tumors. PMID- 12087513 TI - Endoscopic transnasal approach to the pituitary lesions using a navigation system (InstaTrak system): technical note. AB - The endoscopic transnasal approach has become a procedure of choice for the surgical management of pituitary lesions. However, in conventional endoscopic transnasal surgery, the surgeon may become disorientated to the actual operating position. In our series, 31 patients have undergone an endoscopic transnasal approach to the pituitary lesions with the use of the navigation system InstaTrak for real-time imaging. This image guidance system proved valuable for anatomic localization during pituitary surgery. We have reduced the average surgical time, and improved patient outcome. As consequence, complications during surgery should decrease and safety should increase. Intraoperative image guidance is expected to have major advantageous effects on pituitary surgery by allowing the surgeon to remove pathology more efficiently. As this system is improved technically and surgeons become more proficient in its use, there should be better postoperative outcomes. PMID- 12087514 TI - Usefulness of silicone plate for sellar floor reconstruction. AB - During the transsphenoidal approach (TSA), the proper sellar floor reconstruction plays an important role in the surgical results. The authors have evaluated the usefulness of a silicone as a substitute for bone splint in sellar floor reconstruction. Silicone plates were used in 10 patients who had 7 pituitary adenomas, 2 Rathke's cleft cysts, and 1 metastatic tumor. Among the ten cases, seven underwent standard TSAs and three received extended TSAs. The silicone plate was cut to a size slightly larger than that of bone window, inserted with a three-pronged fork, and then adjusted precisely. In nine patients, complications that related to surgical repair did not occur. A postoperative cerebrospinal fluid (CSF) rhinorrhea occurred in one pituitary macroadenoma, and no infection was observed in any case. From the authors' experience, advantages of the silicone plate are its simplicity to shape for any size of defects, and the easy detectability of the previous bone window at reoperation. PMID- 12087515 TI - Genome scan for loci involved in cleft lip with or without cleft palate, in Chinese multiplex families. AB - Cleft lip with or without cleft palate (CL/P) is a common congenital anomaly. Birth prevalences range from 1/500 to 1/1,000 and are consistently higher in Asian populations than in populations of European descent. Therefore, it is of interest to determine whether the CL/P etiological factors in Asian populations differ from those in white populations. A sample of 36 multiplex families were ascertained through probands with CL/P who were from Shanghai. This is the first reported genome-scan study of CL/P in any Asian population. Genotyping of Weber Screening Set 9 (387 short tandem-repeat polymorphisms with average spacing approximately 9 cM [range 1-19 cM]) was performed by the Mammalian Genotyping Service of Marshfield Laboratory. Presented here are the results for the 366 autosomal markers. Linkage between each marker and CL/P was assessed by two-point and multipoint LOD scores, as well as with multipoint heterogeneity LOD scores (HLODs) plus model-free identity-by-descent statistics and the multipoint NPL statistic. In addition, association was assessed via the transmission/disequilibrium test. LOD-score and HLOD calculations were performed under a range of models of inheritance of CL/P. The following regions had positive multipoint results (HLOD > or =1.0 and/or NPL P< or =.05): chromosomes 1 (90-110 cM), 2 (220-250 cM), 3 (130-150 cM), 4 (140-170 cM), 6 (70-100 cM), 18 (110 cM), and 21 (30-50 cM). The most significant multipoint linkage results (HLOD > or =2.0; alpha=0.37) were for chromosomes 3q and 4q. Associations with P< or =.05 were found for loci on chromosomes 3, 5-7, 9, 11, 12, 16, 20, and 21. The most significant association result (P=.009) was found with D16S769 (51 cM). PMID- 12087516 TI - Practice guidelines for the diagnosis and management of group A streptococcal pharyngitis. Infectious Diseases Society of America. PMID- 12087517 TI - Diagnosis of strep throat in adults: are clinical criteria really good enough? AB - The clinical manifestations of group A streptococcal and nonstreptococcal pharyngitis overlap quite broadly. For this reason, the updated Infectious Diseases Society of America practice guideline for group A streptococcal pharyngitis, published in this issue of Clinical Infectious Diseases, recommends laboratory confirmation of the clinical diagnosis by means of either throat culture or a rapid antigen detection test. However, a recently published guideline, developed by a subcommittee of the American College of Physicians American Society of Internal Medicine (ACP-ASIM) in collaboration with the Centers for Disease Control and Prevention, advocates use of a clinical algorithm alone, in lieu of microbiologic testing, for confirmation of the diagnosis in adults for whom the suspicion of streptococcal infection is high. In this discussion, we examine the assumptions of the ACP-ASIM guideline, question whether its recommendations will achieve the stated objective of dramatically decreasing excess antibiotic use, and suggest that its recommendations be confirmed by clinical trials before clinicians abandon long-held teachings regarding diagnosis and management of group A streptococcal pharyngitis. PMID- 12087518 TI - Effect of penicillin resistance of Streptococcus pneumoniae on the presentation, prognosis, and treatment of pneumococcal endocarditis in adults. AB - We performed a clinical study of pneumococcal endocarditis (PE) in adults at 15 major Spanish hospitals during a 21-year period (1978-1998). During this time, 63 patients had PE due to Streptococcus pneumoniae diagnosed. Of the 63 isolates recovered from these patients, 24 (38%) and 6 (10%) showed resistance to penicillin (minimum inhibitory concentration [MIC], 0.1-4 microg/mL) and cefotaxime (MIC, 1 microg/mL), respectively. Twenty-two (35%) of the patients died. Left-side heart failure, but not penicillin resistance, was independently associated with a higher risk of death (odds ratio, 1.33; 95% confidence interval, 1.04-1.71; P=.026). Patients without meningitis who had PE due to penicillin-resistant S. pneumoniae could be treated with high-dose penicillin or a third-generation cephalosporin if the MIC for penicillin was < or =1 microg/mL. For patients with concurrent meningitis, high doses of cefotaxime could be used if the MIC for cefotaxime was < or =1 microg/mL. Early recognition of heart failure and surgery may help to decrease mortality. PMID- 12087519 TI - Broad resistance due to plasmid-mediated AmpC beta-lactamases in clinical isolates of Escherichia coli. AB - Escherichia coli that produce plasmid-mediated AmpC beta-lactamases are rare in the United States. The clinical features associated with infection with these organisms have not been well described. We identified 2 clinical isolates of E. coli that produced the plasmid-mediated AmpC enzyme beta-lactamase CMY-2. These organisms were recovered from urine specimens and were resistant to ceftazidime, ceftriaxone, and cefepime. One isolate was resistant to ertapenem but susceptible to imipenem and meropenem; the other was susceptible to imipenem, meropenem, and ertapenem. One of the 2 infected patients did not require specific therapy; the other required imipenem for cure. The presence of the CMY-2 beta-lactamase was confirmed by DNA sequencing. Hybridization studies confirmed that the bla(CMY-2) gene was on a plasmid in both isolates; in one of them, the probe also hybridized with chromosomal DNA. Infection with plasmid-mediated AmpC beta-lactamases in E. coli in the United States may be associated with treatment failure, and these strains may become a serious nosocomial threat. PMID- 12087520 TI - Use of molecular techniques to distinguish between treatment failure and exogenous reinfection with Mycobacterium tuberculosis. AB - We investigated the means by which drug resistance emerges among drug-susceptible Mycobacterium tuberculosis strains during antituberculosis therapy. Patients who experienced failure of treatment for active pulmonary tuberculosis, who initially received diagnoses of infection with drug-susceptible M. tuberculosis, and who had had at least 3 isolates tested for drug susceptibility were selected from a 6 year period in the Estonian National Reference Laboratory archive. Eleven patients from whom 35 sequential isolates of M. tuberculosis had been obtained were recruited into the study. Their clinical data and treatment charts were analyzed and correlated with drug-susceptibility patterns and IS6110 restriction fragment-length polymorphism (RFLP) profiles. Six patients excreted isogenic drug susceptible M. tuberculosis strains, whereas, in the other 5 patients, the isolated strain shifted from a susceptible to a resistant phenotype. In all cases, this shift correlated to a shift in RFLP pattern, which showed reinfection with a new strain. Exogenous reinfection with drug-resistant M. tuberculosis may be misinterpreted as the emergence of drug resistance if molecular testing techniques are not used. PMID- 12087521 TI - Low serum procalcitonin level accurately predicts the absence of bacteremia in adult patients with acute fever. AB - The ability of measurement of serum procalcitonin (PCT) levels to differentiate bacteremic from nonbacteremic infectious episodes in patients hospitalized for community-acquired infections was assessed. Serum samples were obtained from adult inpatients with fever to determine the serum PCT level, C-reactive protein (CRP) level, and erythrocyte sedimentation rate (ESR). Of 165 patients, 22 (13%) had bacteremic episodes and 143 (87%) had nonbacteremic episodes. PCT levels, CRP levels, and ESRs were significantly higher in bacteremic patients than in nonbacteremic patients (P<.001,.007, and.024, respectively). The best cutoff value for PCT was 0.4 ng/mL, which was associated with a negative predictive value of 98.8%. Area under the receiver operating characteristic curve was 0.83 for PCT, which was significantly higher than that for CRP (0.68; P<.0001) and ESR (0.65; P<.05). A serum PCT level of <0.4 ng/mL accurately rules out the diagnosis of bacteremia. The use of PCT assessment could help physicians limit the number of blood cultures to be processed and the number of antibiotic prescriptions. PMID- 12087522 TI - Effectiveness of acellular pertussis vaccine assessed by hospital-based active surveillance in Germany. AB - We assessed the effectiveness of complete and partial pertussis vaccination in Germany--a country where acellular vaccine is predominantly used--for the prevention of cases of pertussis requiring hospitalization. Vaccine effectiveness was estimated by means of a screening method. Vaccine coverage of children born during the period of June 1996 through December 1998 was assessed by a telephone survey. Data from hospitalized children with pertussis in 1997-1998 and from patients with pertussis complications in 1997-2000 were acquired by a nationwide, hospital-based, active surveillance system. Age-adjusted vaccine effectiveness of completed primary vaccination was estimated to be 99.8% (95% confidence interval [95% CI], 98.9-100). After receipt of 1 dose of vaccine, vaccine effectiveness was as high as 68.0% (95% CI, 45.6-81.1), increasing to 91.8% (95% CI, 84.7-95.7) after receipt of the second dose. Vaccine effectiveness was even slightly higher for pertussis with complications. Thus, even after partial vaccination, acellular pertussis vaccine is highly effective in preventing hospitalizations for pertussis. PMID- 12087523 TI - Efficacy of intranasal virosomal influenza vaccine in the prevention of recurrent acute otitis media in children. AB - To evaluate the efficacy of an intranasal, inactivated, virosomal subunit influenza vaccine for prevention of new episodes of acute otitis media (AOM) in children with recurrent AOM, 133 children aged 1-5 years were randomized to receive the vaccine (n=67) or no vaccination (n=66). During a 6-month period, 24 (35.8%) vaccine recipients had 32 episodes of AOM; 42 (63.6%) control subjects had 64 episodes. The overall efficacy of vaccination in preventing AOM was 43.7% (95% confidence interval, 18.6-61.1; P=.002). Children vaccinated before influenza season had a significantly better outcome than did those vaccinated after the onset of influenza season. The cumulative duration of middle ear effusion was significantly less in vaccinated children than in control subjects. Data suggest that the intranasal virosomal influenza vaccine might be considered among the options for the prevention of AOM in children <5 years old with recurrent AOM. PMID- 12087524 TI - Burden of encephalitis-associated hospitalizations in the United States, 1988 1997. AB - Analysis of the National Hospital Discharge Survey data for 1988-1997 revealed a substantial disease burden associated with encephalitis in the United States: on average, there were nearly 19,000 hospitalizations (7.3 hospitalizations per 100,000 population), 230,000 hospital days, and 1400 deaths annually. For most encephalitis-associated hospitalizations (59.5%), the etiologic agent was unknown or not recorded; the most common etiologic categories among known causes were "viral" (38.2%) and "other infectious" (34.1%). The most common individual diagnoses with known agents were herpetic and toxoplasmic encephalitides (each associated with an annual average of 2100 hospitalizations). Human immunodeficiency virus infection was listed among discharge diagnoses for 15.6% of hospitalizations. Rates of encephalitis-associated hospitalization were highest for children <1 year old and persons > or =65 years old. The etiology of encephalitis was unknown for persons > or =65 years old significantly more often than it was for younger persons. The average cost of an encephalitis-associated hospitalization, as determined by the Healthcare Cost and Utilization Project for 1997, was $28,151, for an annual national cost of hospitalization of $650 million. PMID- 12087525 TI - Japanese encephalitis vaccine for travelers: exploring the limits of risk. AB - The prevention of Japanese encephalitis in travelers presents the juxtaposition of 4 factors: a disease that is widespread throughout Asia, a disease with a low incidence in travelers, a vaccine about which there are safety concerns, and a clinical course that can result in death or permanent disability in two-thirds of symptomatic cases. Travel medicine practitioners often seem to be polarized into 2 groups: a group that gives more weight to the severity of the disease (and therefore often recommend vaccination) and another group that is more persuaded by the low occurrence of cases in travelers (and therefore rarely recommend vaccination). This review assesses the known risks of contracting Japanese encephalitis and the risks associated with the vaccine and tries to develop an appropriate way to recommend this vaccine to travelers who may be at significant risk. PMID- 12087526 TI - Prospective study of measles in hospitalized, human immunodeficiency virus (HIV) infected and HIV-uninfected children in Zambia. AB - Measles in persons coinfected with human immunodeficiency virus (HIV) has been reported to be unusual in its presentation and frequently fatal. To determine the effect of HIV coinfection on the clinical features and outcome of measles, a prospective study of hospitalized children with measles was conducted between January 1998 and October 2000 in Lusaka, Zambia. One-sixth (17%) of 546 children hospitalized with laboratory-confirmed measles were coinfected with HIV. One third of the HIV-infected children hospitalized with confirmed measles were <9 months old, compared with 23% of HIV-uninfected children (P=.03). Few differences in clinical manifestations, complications, or mortality were found between HIV infected and HIV-uninfected children with measles. HIV-infected children constitute a significant proportion of children hospitalized with measles in countries with high HIV prevalence and are more likely to be younger than the age for routine measles immunization. PMID- 12087527 TI - Concomitant risk factors in reports of torsades de pointes associated with macrolide use: review of the United States Food and Drug Administration Adverse Event Reporting System. AB - In this case series, we examined concomitant risk factors mentioned in reports of torsades de pointes, a rare ventricular arrhythmia, that occurred in association with administration of macrolide antimicrobials (e.g., azithromycin, clarithromycin, dirithromycin, and erythromycin). Increasing age, female sex, and concomitant diseases and drug administration believed to increase risks for torsades de pointes were commonly reported. PMID- 12087528 TI - Juvenile human T lymphotropic virus type 1-associated myelopathy. AB - We report the cases of 5 adolescents with human T lymphotropic virus type 1 associated myelopathy/tropical spastic paraparesis, acquired in all but 1 case from the mother. The first symptom in all patients was difficulty in running, which was present for many years before the final diagnosis was made. Follow-up showed an indolent progression, regardless of treatment strategy. PMID- 12087529 TI - A novel, multiple drug-resistant, serotype 24F strain of Streptococcus pneumoniae that caused meningitis in patients in Naples, Italy. AB - Three adult patients in Naples, Italy, had meningitis due to multiple drug resistant serotype 24F Streptococcus pneumoniae isolates. The 3 isolates were genetically indistinguishable and shared pbp2b and pspA sequence types with previously characterized penicillin-resistant clones. This serotype 24F strain was found to be the same clonal type as a previously characterized, penicillin resistant serotype 14 strain. The novel strain has probably arisen through transformation of a serotype 14 strain with type 24F capsular biosynthetic operon sequences. PMID- 12087530 TI - Why the AUC/MIC ratio should not be used to predict the effects of beta-lactams. PMID- 12087531 TI - Human host defense and cytokines in mycobacterial infectious diseases: interleukin-18 cannot compensate for genetic defects in the interleukin-12 system. PMID- 12087532 TI - More experience, but not a new experience. PMID- 12087533 TI - Pulmonary tuberculosis due to multidrug-resistant Mycobacterium bovis in a healthy host. PMID- 12087534 TI - Intradermal regimens for rabies postexposure prophylaxis: more confusion. PMID- 12087536 TI - Is there hope for the prevention of future antimicrobial shortages? PMID- 12087538 TI - Nosocomial bloodstream infections in Finnish hospitals during 1999-2000. AB - Prospective laboratory-based surveillance in 4 Finnish hospitals during 1999-2000 identified 1477 cases of nosocomial bloodstream infection (BSI), with an overall rate of 0.8 BSIs per 1000 patient-days. Of BSI cases, 33% were in patients with a hematological malignancy and 15% were in patients with a solid malignancy; 26% were in patients who had undergone surgery preceding infection. Twenty-six percent of BSIs were related to intensive care, and 61% occurred in patients with a central venous catheter. Sixty-five percent of the 1621 causative organisms were gram positive, 31% were gram negative, and 4% were fungi. The most common pathogens were coagulase-negative staphylococci (31%), Escherichia coli (11%), Staphylococcus aureus (11%), and enterococci (6%). Methicillin resistance was detected in 1% of S. aureus isolates and vancomycin resistance in 1% of enterococci. The 7-day case-fatality ratio was 9% and was highest for infections caused by Candida (21%) and enterococci (18%). The overall rate of nosocomial BSIs was similar to rates in England and the United States, but S. aureus, enterococci, and fungi were less common in our study, and the prevalence of antibiotic resistance was lower. PMID- 12087539 TI - Brevibacterium endocarditis: a first report. AB - There are few case reports of infections caused by Brevibacterium species, and there have been no previously reported cases of endocarditis caused by any of the 6 known species of Brevibacterium. We report the first case of Brevibacterium endocarditis (caused by Brevibacterium otitidis) in a patient with prosthetic heart valves. The patient responded to 6 weeks of treatment with vancomycin and 2 weeks with gentamicin, and she has been receiving long-term maintenance therapy with oral azithromycin. PMID- 12087540 TI - Chronic disseminated Trichosporon asahii infection in a leukemic child. AB - We report a case of chronic disseminated Trichosporon asahii infection in a leukemic child. Administration of amphotericin B lipid complex resulted in rapid control and improvement of the initial infection but failed to prevent the development of chronic disseminated disease. Cure was achieved after treatment was changed to a 20-month course of itraconazole. This case report demonstrates that Trichosporon species can cause chronic disseminated disease with a pattern similar to that of disease caused by Candida species. PMID- 12087541 TI - Absence of human immunodeficiency virus infection after intentional injection of infected blood. AB - We report the case of a young man who injected himself intravenously with 2 mL of human immunodeficiency virus (HIV)-infected blood and failed to develop HIV infection. PMID- 12087542 TI - The response was overwhelming--hopefully it's just the beginning. PMID- 12087543 TI - The dimensions of mandibular lingual tissues relative to the placement of a lingual bar major connector. AB - PURPOSE: The purpose of this investigation was to measure and describe the lingual tissues of the anterior mandible to determine the acceptable boundaries of the zone of nonmovable tissue for placement of a lingual bar and to compare these to existing numerical guidelines. MATERIALS AND METHODS: Eighty subjects, grouped by age and gender, with clinically normal mandibular lingual gingival tissues from second premolar to second premolar were examined. The lingual sulcular depths (AB) and the distance from the gingival crest to movable tissue of the floor of the mouth (AC) were recorded. The zone of lingual nonmovable tissue was calculated (AC - AB) for each tooth. RESULTS: For all subjects, the mean value of the probing depth (AB) ranged from 1.22 mm (+/-0.33 mm) for central incisors to 1.66 mm (+/-0.43 mm) for second premolars. The mean distance from the gingival crest to movable tissue of the floor of the mouth (AC) ranged from 7.44 mm (+/-1.59 mm) for central incisors to 10.28 mm (+/-2.55 mm) for second premolars. The mean height of the lingual nonmovable tissue (the zone available for the lingual bar; AC - AB) ranged from 6.22 mm (+/-1.59 mm) for central incisors to 8.63 mm (+/-2.57 mm) for second premolars. Most subjects presented with the minimum zone of nonmovable tissue at the central incisors, but with increasing age more subjects presented with minimum values at the posterior teeth. The multifactorial analysis of variance (ANOVA) for males' available nonmovable tissue shows significance for both tooth (p <0.001) and age (p <0.001) factors. The multifactorial ANOVA for females' available nonmovable tissue shows significance for both tooth (p <0.001) and age (p <0.001) factors. Of all subjects, 85% (88% of males, 83% of females) had 4 mm or more of nonmovable tissue, sufficient for a lingual bar. The amount of available room decreased in older females. CONCLUSION: Within the limits of this study, the use of actual measurements of lingual tissues versus existing numerical guidelines increased the percentage of patients for which the lingual bar can be used from approximately 6% to approximately 85%. PMID- 12087544 TI - Effects of carbamide peroxides on wear resistance and color change of enamel opposing porcelain. AB - PURPOSE: This study evaluated the effects of a simulated 1- and 2-week night guard bleaching technique using various bleaching gel concentrations on wear resistance and color change of human enamel opposing a feldspathic porcelain substrate. MATERIALS AND METHODS: VMK 68 feldspathic porcelain was formed into 15 mm disks (n = 63) and fired according to manufacturer's recommendations. Enamel cusps were formed and shaped from extracted third molars and treated with 1 of 3 bleaching materials for 7 or 14 days. Specimens were kept at 37 degrees C in 100% humidity. The control group was subjected to identical conditions but with water replacing the bleaching gel between and during bleaching treatments. After simulated bleaching, the enamel cusp specimens were subjected to 3-body wear testing opposite a highly polished porcelain substrate. Enamel wear was evaluated using optical scanning methods, and color was measured with a small-aperture colorimeter. Data were subjected to a one-way repeated- measure analysis of variance and various post hoc statistics. RESULTS: Results indicated that the within effect of treatment time (7 or 14 days) did not significantly affect (p = 0.13) the size of the resulting enamel wear facet, whereas the between effect of bleaching concentration was significant (p = 0.01). Statistical analysis of unbleached control groups suggests that between-group differences can be explained by variations in wear rates of the 3 enamel groups, not by the concentration differences in bleaching gels. All bleaching treatments resulted in significant color changes (p = 0.04) compared to the unbleached control, but the bleach concentration did not significantly influence the magnitude of color change (p = 0.81). CONCLUSIONS: A simulated night guard vital bleaching treatment of 1 or 2 weeks had no significant effect on the wear resistance of human enamel in vitro. The increased bleaching treatment time and concentration did not significantly affect the amount of enamel color change. PMID- 12087545 TI - Laboratory strength of glass ionomer cement, compomers, and resin composites. AB - PURPOSE: The study evaluates the compressive, flexural, and diametral tensile strengths of 8 core build-up materials from different material classes (highly viscous glass ionomer cement, autocured resin composite, and compomers). MATERIALS AND METHODS: All materials were manipulated according to the manufacturers' recommendations for use as core materials. At a temperature of 23.0 +/- 1.0 degrees C the properties of compressive, diametral tensile and flexural strength were determined using a universal testing machine at 15 minutes, 1 hour, and 24 hours after material preparation. Using one-way analysis of variance (ANOVA), multiple mean value comparisons were performed to determine significant differences (p< or =.05) between the core restoration materials. RESULTS: The values for compressive strength varied from 40.3 +/- 5.2 MPa (compomer) to 237.4 +/- 37.3 MPa (autocured resin composite) for the 3 measurement times. At 15 minutes, 1 hour, and 24 hours after first mixing, the ANOVA showed significant differences (p < or =.05) between the resin composite Core Paste and all of the other materials. Diametral tensile strengths ranged from 5.5 +/- 1.1 MPa for glass ionomer cement to 39.1 +/- 2.9 MPa for composite core material. Three-point flexural strength showed values ranging from 12.1 +/- 2.5 MPa for glass ionomer cement to 92.1 +/- 9.7 MPa for compomer between the 3 measurement times. CONCLUSIONS: Setting time influences the mechanical properties of the materials tested in this study. Autopolymerizing resin composite Core Paste demonstrated greater compressive and flexural strengths at the 3 measurement times than the other materials tested. Reinforced composites, in comparison to the autocured resin composites, yielded no improvement in tensile strength. Flexural and tensile strengths of the glass ionomer cement were lower than those of autocured resin composites and compomers. PMID- 12087546 TI - Effect of three endodontic materials on the bond strength of two composite core materials to dentin. AB - PURPOSE: This study investigated the effect of dentin treatment with iodine potassium iodide, calcium hydroxide or a biphenol-A- diglycidylether epoxy resin sealer on the dentin bond strength of titanium-reinforced Bis-GMA and urethane dimethacrylate composite materials. MATERIALS AND METHODS: Dentin was exposed in four groups of extracted teeth (20 specimens each). Three groups were treated with one of the contaminants and the fourth group was left uncovered and served as the control group. All specimens were stored for 1 week at 37 degrees C and 100% humidity. The dentin surfaces were cleaned with pumice slurry using a prophylaxis rubber cup, rinsed and dried. Then the dentin surfaces were treated using the conditioner and the primer of the GLUMA adhesive system followed by placement of composite core materials. After 20 minutes, debonding of the core materials was accomplished using a shear-peel test. A two-way analysis of variance and Tukey's multiple range test was performed based upon core type and contaminant type (alpha = 0.05). RESULTS: Titanium-reinforced Bis-GMA composite showed significantly greater bond strength values than the urethane dimethacrylate composite material (p < 0.0001). The mean bond strength values in megapascals for the Bis-GMA resin ranged from 8.47 +/-1.52 for the calcium hydroxide group to 9.81 +/-0.94 for the control group. Mean bond strengths for the urethane dimethacrylate resin ranged from 3.35 +/-0.90 for calcium hydroxide to 3.99 +/-0.88 for iodine potassium iodide groups. Within each core group, no difference in bond strengths were noted compared to the uncontaminated control. CONCLUSION: Pretreatment of the dentin with iodine potassium iodide, calcium hydroxide, or epoxy resin followed by pumicing and using the GLUMA system conditioner and primer had no effect on the bond strength of the two resin composite core materials. However, the titanium reinforced Bis-GMA composite exhibited significantly greater bond strength to dentin than the urethane dimethacrylate based composite. PMID- 12087547 TI - Dimensional accuracy of dental casts: influence of tray material, impression material, and time. AB - PURPOSE: This study evaluated the influence on dimensional accuracy of dental casts made with different types of trays and impression materials and poured at different and multiple times. MATERIALS AND METHODS: Two types of stock trays (plastic stock tray, perforated metal stock tray) and 4 types of custom tray materials (autopolymerizing acrylic resin, thermoplastic resin, and 2 types of light-polymerized acrylic resins) were used with 2 types of impression materials (addition polymerizing silicone and polyether), to make impressions of a metal master model. Each tray and impression material was used to make 5 impressions. Casts were made by multiple pourings at 30 minutes, 6 hours, 24 hours, and 30 days after impression making. Using a measuring microscope, 12 distances were calculated based on measurements of 8 reference points. The absolute value of the difference of each measurement was calculated, as was the corresponding measurement on the master model. A Bayesian model using a simple noninformative prior was used to analyze these data. RESULTS: Statistical differences within 6 microm were found only with thermoplastic resin tray material for addition silicone, and for thermoplastic resin tray material and 1 type of light polymerized acrylic resin for polyether. Neither stock trays nor custom trays contributed to the differences in accuracy of the casts. All deviations in casts made with silicone impression material were within a clinically acceptable range. For the polyether, distortions occurred that were clinically unacceptable. Impressions made from polyether distorted over time. Silicone impression material has dimensional stability up to 30 days. CONCLUSION: Accurate casts can be made with either stock trays or custom trays. An impression made from polyether should be poured only once and within 24 hours after impression making, because of the distortion of the material over time. Silicone impression material has better dimensional stability than polyether. . PMID- 12087548 TI - Effect of opacifiers on color stability of pigmented maxillofacial silicone A 2186 subjected to artificial aging. AB - PURPOSE: This study was designed to determine the effect of opacifiers used at different ratios on the color stability of pigmented A-2186 silicone maxillofacial elastomers and to evaluate the color spectrophotometrically before and after artificial aging. MATERIALS AND METHODS: Sixty experimental groups of elastomers were made using various concentrations (5%, 10%, and 15%) of 4 opacifiers (Georgia kaolin powder neutral, kaolin powder calcined, Artskin white, and dry pigment titanium white) with 1 of 5 dry earth cosmetic pigment groups (no pigment [control], red, yellow ochre, burnt sienna, and a mixture of all pigments). Five specimens of each elastomer were tested, for a total of 300 specimens. All specimens were placed in an aging chamber and artificially aged by exposure to light, water spray, fluctuating temperatures, and humidity. CIE L*a*b* values were measured by spectrophotometer. The color differences (Delta E*) at various exposure energies (150, 300, and 450 kJ/m(2)) were subjected to 4 way analysis of variance with repeated measures (super ANOVA). Mean values were compared with Tukey-Kramer intervals calculated at the 0.05 significance level. RESULTS: The trained human eye can detect color changes (Delta E*) greater than 1.0. Adding all pigments to any of the kaolin groups did not protect silicone A 2186 from color degradation over time. Mixing red pigment in all groups at all times drastically increased DeltaE* values ranging from 0.1 to 1.3 up to 16.6 to 49.6. Yellow ochre had an effect only with 10% and 15% concentrations of kaolin powder calcined, increasing the value of Delta E* to greater than 1.0. Burnt sienna had an effect only with a 15% concentration of kaolin powder calcined, increasing the value of Delta E* at 300 and 450 kJ/m(2) to greater than 11.0. At the 5% concentration, kaolin powder calcined had the smallest color changes, followed by, in order, dry pigment titanium white, Artskin white, and Georgia kaolin. At the 10% concentration, Artskin white had the smallest color changes, followed by, in order, dry pigment titanium white, kaolin powder calcined, and Georgia kaolin. At the 15% concentration, Artskin white again had the smallest color changes, followed by, in order, dry pigment titanium white, Georgia kaolin, and the kaolin powder calcined. The smallest color changes in each kaolin group were at the 10% concentration for Artskin white, dry pigment titanium white, and kaolin powder calcined and at the 5% concentration for Georgia kaolin. CONCLUSIONS: Mixing dry earth cosmetic pigments with opacifiers did not protect silicone A-2186 from color degradation over time, especially in the case of red pigment. The group in which pigments were mixed with 10% Artskin white had the smallest color changes over time, followed by, in order, the groups in which pigments were mixed with 10% dry pigment titanium white, 10% kaolin powder calcined, and 5% Georgia kaolin. Red pigment had a significant effect on all opacifiers, especially Georgia kaolin and kaolin powder calcined. Yellow ochre and burnt sienna had an effect only on 15% kaolin powder calcined. Among all the pigment groups tested, yellow ochre remained the most color stable over time. PMID- 12087549 TI - Cast articulation accuracy using rigid cast stabilization. AB - PURPOSE: This study evaluated the positional accuracy of casts articulated on a semi-adjustable articulator, with and without rigid cast stabilization using either laboratory plaster or mounting plaster. MATERIALS AND METHODS: A reference articulation of melamine casts in maximum articulation was established and recorded in the horizontal and vertical dimensions using a verification device. The same casts were subsequently remounted 24 times using either laboratory plaster or mounting plaster. Half of the articulations from each group were stabilized using detachable mounting rods and sticky wax, and half were hand articulated without stabilization, for a total of 6 articulations in each of 4 test groups. The resulting spatial positions established on the articulator were compared to the initial reference position on the verification device grid. Means and standard deviations of the absolute values of the horizontal and vertical displacement for each group were determined separately and compared using a one way anaylsis of variance. Significant differences (p <0.05) were identified using Tukey's honestly significant difference multiple comparison test. RESULTS: Mean vertical mandibular cast displacement ranged from 0.26 +/-0.21mm for stabilized casts mounted with laboratory plaster to 1.58 +/-0.32 mm for unstabilized casts mounted with mounting plaster. For each mounting material, significantly less vertical displacement (p <0.001) was observed with the mandibular cast stabilized before mounting. The cast mounted with laboratory plaster exhibited horizontal displacement (0.87 +/-0.29 mm) that was significantly greater than the remaining groups (p <0.001), which did not differ from each other. CONCLUSIONS: Rigid stabilization of the mandibular to maxillary cast during mounting with laboratory and mounting plaster improved articulation accuracy. PMID- 12087550 TI - The history and development of the dental surveyor: part II. AB - Part II of this series on the history and development of the dental surveyor reviews ways in which the basic dental surveyor has been modified to improve its accuracy and versatility. . PMID- 12087551 TI - Photographing a cast metal restoration. PMID- 12087552 TI - The history of articulators: a critical history of articulators based on geometric theories of mandibular movement: part I. PMID- 12087554 TI - Association between high-normal albuminuria and risk factors for cardiovascular and renal disease in essential hypertensive men. AB - BACKGROUND: Microalbuminuria (overnight urinary albumin excretion [UAE] > 15 microg/min) is associated with cardiovascular risk factors and predicts morbid events in hypertensive subjects. However, albuminuria is not a dichotomous variable, and a relationship with cardiovascular risk factors may extend below that conventional threshold. METHODS: We studied 186 never-treated, glucose tolerant, normalbuminuric (overnight UAE < or = 15 microg/min), essential hypertensive men with normal renal function (serum creatinine < 1.4 mg/dL). Study variables were 24-hour ambulatory blood pressure (BP), cardiac structure and geometry (by echocardiography), body weight, fasting insulin levels, insulin sensitivity (the Homeostasis Model Assessment index), and creatinine clearance (from overnight collections or through the Cockcroft formula) analyzed as a function of ascending urine albumin quartiles (cutoff values, 4.3, 6.3, and 9.4 microg/min; n = 47, 45, 47, and 47, respectively). RESULTS: As compared with the three bottom fourths, patients with high-normal albuminuria (albumin, 9.4 to 15 microg/min) had a greater 24-hour BP, greater relative wall thickness, more frequent concentric left ventricular hypertrophy, heavier body size, increased fasting insulin levels, reduced insulin sensitivity, and greater creatinine clearance. CONCLUSIONS: High-normal albuminuria in uncomplicated essential hypertensive men is associated with an adverse cardiovascular and metabolic risk profile. Furthermore, hyperfiltration in the presence of minimally increased albuminuria may underlie an augmented glomerular blood flow and hydraulic pressure conducive to glomerular hypertension and, eventually, renal insufficiency. Overall, these data confirm the appropriateness to shift downward the limits for diagnosing microalbuminuria in essential hypertension, as indicated from previous prospective studies. PMID- 12087556 TI - Recurrent fetal loss associated with bilineal inheritance of type 1 autosomal dominant polycystic kidney disease. AB - BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is a common Mendelian disorder that affects approximately 1 in 500 to 1,000 live births. Mutations in one of two genes, PKD1 and PKD2, account for the disease in most ADPKD families. Despite the relative high frequency of PKD1 mutant alleles, compound heterozygotes or diseased homozygotes have not been described. METHODS AND RESULTS: We report a family with type 1 ADPKD in which the marriage between affected first-degree cousins resulted in two live-born heterozygous offspring and two fetuses lost in late pregnancy. Genetic analysis with PKD1 and PKD2 flanking markers showed that this family is PKD1 linked (z(max) = 1.66 and -2.54 at thetas = 0.0 for intragenic markers for PKD1 [ie, KG8] and PKD2 [ie, SPP1], respectively). CONCLUSION: Given a 25% chance for mutant homozygosity in the offspring of this family, our findings suggest that homozygosity of PKD1 mutations in humans is embryonically lethal, as recently documented in Pkd1 knockout mice. PMID- 12087555 TI - Atrial natriuretic peptide-induced microalbuminuria is associated with endothelial dysfunction in noncomplicated type 1 diabetes patients. AB - BACKGROUND: Microalbuminuria reflects widespread vascular dysfunction in type 1 diabetes mellitus and results from increased glomerular sieving caused by changes in transglomerular pressure and/or permselectivity characteristics of the glomerular basement membrane. Increased tubular reabsorption or degradation of albumin will offset an early increase in albuminuria. We hypothesized that the infusion of atrial natriuretic peptide (ANP) as a tool to increase glomerular permeability might uncover changes in permselectivity in patients with uncomplicated type 1 diabetes. METHODS: We investigated whether these patients were characterized by endothelial and/or vascular dysfunction. We therefore studied 46 normoalbuminuric patients (urinary albumin excretion [UAE] < 10 microg/min) with type 1 diabetes and 44 healthy controls. Measurements of renal hemodynamics and albuminuria were performed before (baseline) and during the infusion of ANP (0.01 microg/kg/min). On a separate occasion, endothelial function was assessed by the intra-arterial infusion of acetylcholine (ACh), an endothelial-dependent vasodilator. RESULTS: At baseline, glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) were greater in patients with diabetes (GFR, 121 +/- 3 versus 106 +/- 2 mL/min/1.73 m(2); ERPF, 558 +/- 16 versus 527 +/- 13 mL/min/1.73 m(2); P < 0.001). The infusion of ANP increased filtration fraction. There were no differences in these responses between groups. UAE was significantly greater in patients with diabetes after the ANP infusion (15.8 +/- 1.4 [+183%] versus 9.5 +/- 1.3 microg/min [+96%]; P < 0.01). A subgroup of patients with diabetes with an enhanced albuminuric response (change in UAE > 2 SD of controls) to ANP infusion (mean UAE, 30.3 +/- 1.0 microg/min; 425% +/- 61%) was characterized by a diminished vasodilatory response to ACh (maximal forearm blood flow, 17.2 +/- 2.9 [+563%] versus 26.3 +/- 2.3 mL/min/dL [+800%] in patients with diabetes with a normal albuminuric response; P < 0.05). CONCLUSIONS: In a subgroup of patients with uncomplicated type 1 diabetes, an increase in glomerular permselectivity can be unmasked by the infusion of ANP. These patients are characterized by a diminished vascular response to ACh. PMID- 12087557 TI - Autosomal recessive distal renal tubular acidosis caused by G701D mutation of anion exchanger 1 gene. AB - Anion exchanger 1 (AE1 or band 3), encoded by the AE1 or SLC4A1 gene, regulates chloride-bicarbonate exchange in erythrocytes and alpha-intercalated cells of the distal nephron. Defects of AE1 at the basolateral membrane of alpha-intercalated cells may result in the failure of hydrogen ion secretion at the apical membrane, leading to distal renal tubular acidosis (dRTA). Abnormalities of the AE1 gene were previously reported to be associated with autosomal dominant dRTA. However, recent studies of Thai dRTA families have shown that mutations in this gene result in autosomal recessive (AR) dRTA, giving rise to the postulation that AE1 gene mutations causing AR dRTA might be found commonly in Thai pediatric patients with dRTA. We performed a study of the AE1 gene using DNA linkage, polymerase chain reaction single-strand conformation polymorphism, restriction endonuclease HpaII digestion, and DNA sequence analyses in eight families involving 12 Thai children with dRTA, shown by abnormal urinary acidification using a short acid loading test, as well as among their family members. Seven patients with dRTA from five families had the same homozygous missense G701D mutation of the AE1 gene. Their parents or siblings heterozygous for the AE1 G701D mutation were clinically normal and did not have abnormal urinary acidification, although a heterozygous sibling in one family had abnormal urinary acidification. Results of this and previous studies show that a homozygous AE1 G701D mutation causes AR dRTA and is a common molecular defect among Thai pediatric patients with dRTA. PMID- 12087558 TI - Quality of prereferral care in patients with chronic renal insufficiency. AB - BACKGROUND: Appropriate care in chronic renal insufficiency (CRI) includes blood pressure and diabetes control, as well as the investigation and management of anemia, acidosis, and bone disease. There is a lack of data on the control of these parameters at the time of referral to a nephrologist. Similarly, early referral has been emphasized in the literature, yet very little published has examined current referral patterns. METHODS: A single-center retrospective/prospective review of all new outpatient referrals to nephrologists in Halifax, Canada, in 1998 and 1999 was conducted to identify patients with CRI (serum creatinine > 1.6 mg/dL [141 micromol/L] for men or >1.2 mg/dL [106 micromol/L] for women). Quality of prereferral care was based on data from the initial clinic visit. RESULTS: Of 1,050 charts reviewed, 411 patients met the study criteria. Twenty-six percent of patients had diabetes mellitus, 18% were referred with a calculated glomerular filtration rate less than 15 mL/min, and blood pressure was optimally controlled (<130 mm Hg systolic and <80 mm Hg diastolic) in only 24%. Only 44% of patients were administered an angiotensin converting enzyme inhibitor. Patients were administered an average of 1.9 antihypertensive agents. Significant anemia (hemoglobin < 10 g/dL) was present in 21%, and appropriate investigations were performed in only 35% of these patients. Calcium levels less than 8.6 mg/dL (2.15 mmol/L) were found in 19% of patients, and only 14% of these patients were started on calcium supplement therapy. Phosphate levels greater than 5.0 mg/dL (1.6 mmol/L) were seen in 20% of patients, and 14% of these patients were on phosphate-binder therapy. Parathyroid hormone levels were more than five times normal values in 18% of patients, and 25% of patients had bicarbonate levels less than 23 mmol/L. CONCLUSIONS: A significant proportion of patients referred with CRI receive inadequate prereferral care. Continuing education programs and referral guidelines must not only emphasize the importance of early referral, but also address the related consequences of CRI to delay the progression of renal disease and avoid complications. PMID- 12087559 TI - The association between gout and nephrolithiasis: the National Health and Nutrition Examination Survey III, 1988-1994. AB - BACKGROUND: Gout, an inflammatory arthritis, reportedly afflicts more than 2 million men and women in the United States. Previous reports have suggested an association between gout and kidney stone disease; however, these studies did not adjust for such important potential confounders as obesity and the presence of hypertension. To our knowledge, no published study has examined the independent association between gout and kidney stone disease. METHODS: We used a national probability sample of the US population to determine the independent association between reported gout and history of kidney stone disease. RESULTS: Among men and women 20 years and older, 5.6% (10 million) reported the previous passage of a kidney stone and 2.7% (5.1 million) reported a diagnosis of gout by a physician. Moreover, 8.6% of individuals who reported the passage of a kidney stone on two or more occasions had a history of gout. Conversely, the prevalence of previous kidney stones in subjects with reported gout was 13.9%. In the age-adjusted model, gout was associated with an increased odds ratio (OR) for previous kidney stones (OR, 1.97; 95% confidence interval [CI], 1.37 to 2.83). After further adjustment for sex, race, body mass index, and presence of hypertension, the OR for previous kidney stones in individuals with gout decreased to 1.49 (95% CI, 1.04 to 2.14). CONCLUSION: Showing an independent association between kidney stone disease and gout strongly suggests that they share common underlying pathophysiological mechanisms. Identification of these mechanisms may lead to improved preventive strategies for both conditions. PMID- 12087560 TI - Erythrocyte aminolevulinate dehydratase activity as a lead marker in patients with chronic renal failure. AB - BACKGROUND: Overexposure to lead may result in an increased risk for developing chronic renal failure (CRF) and hypertension. Subclinical lead poisoning is difficult to identify. Because the heme biosynthetic pathway is highly sensitive to lead, we considered the study of enzymes involved in this pathway as a method to detect an excessive body lead burden. METHODS: Main concerns in assessing the heme pathway in patients with CRF were related to aminolevulinate dehydratase (ALAD) activity. We first selected a number of patients with CRF at a predialysis stage, subsequently dividing them into two groups after the EDTA mobilization test had determined whether lead pools were expanded. The study included 24 healthy controls, 12 patients with clinical plumbism and biochemical demonstration of lead poisoning (Pb-CONT), 18 patients with CRF with no evidence of high lead storage (CRF/-), and 8 patients with CRF with high urinary excretion of lead in contrast to normal blood lead levels (CRF/+). RESULTS: As expected, symptoms of plumbism (Pb-CONT) were accompanied by an increased erythrocyte zinc protoporphyrin-free protoporphyrin ratio and high urine coproporphyrin excretion, whereas both these values were within the normal range in all patients with CRF. CRF/- patients showed minor abnormalities of erythrocyte heme metabolism, such as low ALAD activity, both baseline and in vitro restored. The ALAD-restored ALAD ratio correlated closely with urine lead excretion; it was normal in healthy controls and CRF/- patients and significantly reduced in Pb-CONT and CRF/+ patients. CONCLUSION: The erythrocyte ALAD-restored ALAD ratio may be a useful tool to show otherwise subclinical lead poisoning in patients with CRF. PMID- 12087561 TI - Lead mobilization during calcium disodium ethylenediaminetetraacetate chelation therapy in treatment of chronic lead poisoning. AB - It has long been established that chronic lead (Pb) poisoning is a cause of renal insufficiency. However, although easily diagnosed, there is still no treatment available that will revert this type of poisoning. We report a study performed on 56 male Wistar rats administered Pb in drinking water (500 ppm Pb acetate) over a 90-day period. Twenty-one non-Pb-exposed animals served as the control group. Seven animals from each group were killed days 60 and 90. At the end of the 90 day period, 21 of the Pb-exposed animals were treated with disodium monocalcium EDTA (50 mg/kg/d for 5 days) intraperitoneally and 21 animals were administered serum saline by the same route. Three treatment courses were administered, separated by 9 days free of treatment. Seven animals from each subgroup were killed at the end of each treatment course. Pb levels were determined in blood, urine, liver, brain, kidney, and bone. Treatment with EDTA led to a greater and more rapid reduction in Pb contents in the brain and kidney. The decrease in hepatic Pb levels in the treated group of animals was similar to that in the group administered placebo. Bone Pb levels also failed to show a response to the chelating agent. Use of EDTA appears to result in a reduction in Pb deposits in such critical organs as the kidney and brain. However, the chelating agent does not seem to have access to bone Pb deposits, such that the skeleton becomes a permanent source of poisoning for other tissues. PMID- 12087562 TI - Experimental lead nephropathy: treatment with calcium disodium ethylenediaminetetraacetate. AB - Chronic lead poisoning may cause hypertension, gout, and renal insufficiency. Most experimental poisoning studies have involved the use of high doses over short periods (ie, acute poisoning). Although chelating treatment leads to remission of acute lead nephropathy, its effects in the treatment of chronic poisoning are unclear. The aims of this study were to evaluate renal alterations produced during chronic lead poisoning and their progression when poisoning was over and to determine the efficiency of chelating treatment with calcium disodium ethylenediaminetetraacetate (EDTA). In this study, 56 male Wistar rats were administered lead in drinking water (500 ppm lead acetate) over 90 days. The control group consisted of 21 nonexposed rats. Seven rats from each group were killed on days 60 and 90. At the end of the 90-day period, 21 of the lead-exposed rats were treated with disodium monocalcium EDTA (50 mg/kg/d x 5 days) intraperitoneally, and 21 were administered serum saline by the same route. Three treatment courses were given separated by 9 days free of treatment. Seven rats from each subgroup were sacrificed at the end of each treatment course. Main findings related to poisoning were hypertrophy and vacuolization of medium and small arteries; mucoid edema and muscular hypertrophy in arterioles; loss of cell brush borders, cell loss, and intranuclear inclusion bodies in the proximal tubule; and fibrosis and the presence of infiltrates in the interstitial component. Treatment with EDTA slowed the progression of most alterations. No damage associated with the use of the chelating agent was observed. Longer term studies of the effects of this drug are required to establish whether the damage caused by lead poisoning may be reversed. PMID- 12087563 TI - Troponin is related to left ventricular mass and predicts all-cause and cardiovascular mortality in hemodialysis patients. AB - Cardiac troponin T (cTnT) predicts death and cardiovascular outcomes in clinically stable patients with end-stage renal disease. Because this protein is synthesized exclusively in myocardial cells, its predictive power for these outcomes may be because it reflects, besides cardiac ischemia, left ventricular (LV) mass, which is a strong predictor of cardiovascular death in this population per se. We tested the relationship between cTnT level and LV mass and the predictive power of this cardiac protein for all-cause and cardiovascular mortality in a cohort of hemodialysis patients (n = 199) without acute coronary syndrome and heart failure followed up for an average of 35 months (range, 0.8 to 52 months). cTnT was measured by means of a third-generation electrochemiluminescence immunoassay. cTnT level was related directly to interventricular septum (r = 0.36; P < 0.001) and posterior wall thickness (r = 0.40; P < 0.001), as well as LV mass (r = 0.45; P < 0.001). On multivariate analysis, after age, LV mass was the strongest independent predictor of cTnT level (beta = 0.28; P < 0.001). Serum cTnT level was significantly related to all cause and cardiovascular mortality on univariate analysis (P < 0.001). On multivariate Cox regression analysis, the adjusted risk for all-cause death was 2.39 times (95% confidence interval [CI], 1.13 to 5.06; P = 0.02) greater in patients in the third cTnT tertile than the first tertile, and a similar pattern emerged for cardiovascular mortality (hazard ratio, 2.35; 95% CI, 1.01 to 5.49; P = 0.048). In hemodialysis patients, plasma cTnT level is independently related to LV mass and predicts all-cause and cardiovascular mortality. These data support the hypothesis that this marker can be usefully applied for risk stratification in clinically stable dialysis patients. PMID- 12087564 TI - Role of nitric oxide, endothelin-1, and inflammatory cytokines in blood pressure regulation in hemodialysis patients. AB - BACKGROUND: Altered regulation of blood pressure (BP) in hemodialysis patients is associated with increased morbidity and mortality. Regulation of BP is dependent in part on such vasoactive agents as nitric oxide (NO) and endothelin-1 (ET-1). Cytokine-mediated NO synthase activation during dialysis previously has been reported. The purpose of this study is to investigate the relationship between cytokine-mediated activation of the NO and ET-1 systems and BP regulation in hemodialysis patients. METHODS: Nine patients with chronic hypotension (predialysis systolic BP < 110 mm Hg, duration > 1 month), nine patients with hypertension (predialysis systolic BP > or = 180 mm Hg), and nine age- and sex matched normotensive controls were enrolled. RESULTS: Predialysis NO end product levels in the hypotensive group were greater than in controls (17.63 +/- 5.9 versus 11.06 +/- 2.12 microm/mL; P = 0.01), whereas the hypertensive group showed lower levels (4.76 +/- 2.33 microm/mL; P < 0.01). The hypotensive group had low postdialysis levels (3.45 +/- 1.11 microm/mL; P = 0.01). Predialysis ET-1 levels in the hypotensive and hypertensive groups were greater in comparison to the normotensive group (7.54 +/- 4.52 and 8.95 +/- 3.52 versus 4.41 +/- 0.6 pg/mL; P < 0.01). Postdialysis endothelin levels increased in both the control and hypertensive groups (P < 0.01). Interleukin-1 and tumor necrosis factor-alpha levels increased postdialysis in all groups, but not significantly. CONCLUSION: High levels of NO end products in hypotensive patients and low levels in hypertensive patients suggest a critical influence of NO in BP control. In addition, elevated ET-1 levels in hypertensive patients may contribute to systemic vasoconstriction and may suggest vascular dysfunction in this patient population. PMID- 12087565 TI - How to adjust for comorbidity in survival studies in ESRD patients: a comparison of different indices. AB - BACKGROUND: Many patients with end-stage renal disease (ESRD) have additional comorbid conditions. Differences in the presence and severity of these comorbid conditions can bias comparisons between treatment groups. Adjustment for prognostic factors can statistically counterbalance these differences. For this purpose, appropriate weighting of comorbid conditions is necessary. We evaluated three existing methods to score comorbidity in patients with ESRD and compared their ability to predict survival: the Khan, Davies, and Charlson indices. In addition, these three indices were compared with a new index that explicitly incorporates the severity grading of a number of comorbid diseases. METHODS: In a large Dutch prospective multicenter study (Netherlands Co-operative Study on the Adequacy of Dialysis-2), new patients with ESRD were included. Comorbidity was assessed at the start of dialysis therapy. Patient data were randomly allocated to a modeling or testing set. The new index was developed in the modeling set. All indices were evaluated in the testing set. RESULTS: We obtained data for 1,205 patients. Of the three existing indices, the Charlson index had the best discriminating features, with a concordance c statistic of 0.71. The addition of severity grading of several comorbid conditions did not improve discrimination. After combining the comorbidity indices with age, all c statistics improved. These final values ranged from 0.72 to 0.75. CONCLUSION: We conclude that the Khan, Davies, and Charlson scores are appropriate for expressing the prognostic impact of comorbidity on mortality risk in patients with ESRD provided sufficient adjustment for age is performed. Adding the severity grading of several comorbid conditions will not lead to improved prognostic power. PMID- 12087566 TI - Parenteral iron formulations: a comparative toxicologic analysis and mechanisms of cell injury. AB - BACKGROUND: Multiple parenteral iron (Fe) formulations exist for administration to patients with end-stage renal disease. Although there are concerns regarding their potential toxicities, no direct in vitro comparisons of these agents exist. Thus, the present study contrasted pro-oxidant and cytotoxic potentials of four available Fe preparations: Fe dextran (Fe dext), Fe sucrose (Fe sucr), Fe gluconate (Fe gluc), and Fe oligosaccharide (Fe OS). METHODS: Differing dosages (0.06 to 1 mg/mL) of each compound were added to either (1) isolated mouse proximal tubule segments, (2) renal cortical homogenates, or (3) cultured human proximal tubule (HK-2) cells (0.5- to 72-hour incubations). Oxidant injury (malondialdehyde generation) and lethal cell injury (percentage of lactate dehydrogenase release; tetrazolium dye uptake) were assessed. Effects of selected antioxidants (glutathione [GSH], catalase, dimethylthiourea (DMTU), and sodium benzoate also were assessed. RESULTS: Each test agent induced massive and similar degrees of lipid peroxidation. Nevertheless, marked differences in cell death resulted (Fe sucr >> Fe gluc > Fe dext approximately Fe OS). This relative toxicity profile also was observed in cultured aortic endothelial cells. Catalase, DMTU, and sodium benzoate conferred no protection. However, GSH and its constituent amino acid glycine blocked Fe sucr-mediated cell death. The latter was mediated by mitochondrial blockade, causing free radical generation and a severe adenosine triphosphate depletion state. CONCLUSIONS: (1) parenteral Fes are highly potent pro-oxidants and capable of inducing tubular and endothelial cell death, (2) markedly different toxicity profiles exist among these agents, and (3) GSH can exert protective effects. However, the latter stems from GSH's glycine content, rather than from a direct antioxidant effect. PMID- 12087567 TI - Factors contributing to higher hematocrit levels in hemodialysis patients not receiving recombinant human erythropoietin. AB - Recombinant human erythropoietin (rHuEPO) is used to correct anemia in the majority of hemodialysis patients, but a few patients can maintain greater hematocrits without the use of rHuEPO. We aim to investigate which factors stimulate erythropoiesis, other than rHuEPO, in hemodialysis patients. One hundred fifty-eight patients undergoing regular hemodialysis treatment participated in a cross-sectional study. To keep the target hematocrit of 30%, 133 patients (84%) were administered rHuEPO, but 25 patients (16%) did not need rHuEPO. Mean hematocrits were 33.4% +/- 4.6% in patients who did not need rHuEPO and 30.9% +/- 4.0% in those administered rHuEPO. In the analysis of factors contributing to the lack of requirement of rHuEPO with multivariate logistic regression analysis, years on dialysis therapy and body mass index (BMI) were determined to be independent factors with odds ratios of 1.12 (95% confidence interval [CI], 1.02 to 1.23; P = 0.02) and 1.36 (95% CI, 1.13 to 1.63; P = 0.001), respectively. Neither serum erythropoietin level, albumin concentration, nor normalized protein catabolic rate contributed to the lack of requirement of rHuEPO. BMI correlated closely with log serum leptin level (r = 0.55; P < 0.0001), and log serum leptin level correlated inversely with rHuEPO dose (r = 0.18; P = 0.03). These results indicate that 16% of hemodialysis patients could maintain greater hematocrits without the administration of rHuEPO, and independently contributing factors were greater BMI and more years on hemodialysis therapy. Regarding nutritional parameters, a relatively greater BMI, possibly through effects of greater leptin levels, may stimulate erythropoiesis in uremic patients even in the absence of sufficient erythropoietin production. PMID- 12087568 TI - Randomized, controlled trial of darbepoetin alfa for the treatment of anemia in hemodialysis patients. AB - BACKGROUND: Darbepoetin alfa (Aranesp; Amgen, Thousand Oaks, CA) is a new erythropoiesis-stimulating protein with a threefold longer terminal half-life than recombinant human erythropoietin (epoetin) in patients with chronic kidney disease (CKD). The purpose of this randomized, double-blind, noninferiority study is to determine whether darbepoetin alfa is as effective as epoetin for the treatment of anemia in hemodialysis patients when administered at a reduced dosing frequency. METHODS: Patients receiving epoetin therapy were randomized to continue epoetin administered intravenously (IV) three times weekly (n = 338) or change to darbepoetin alfa administered IV once weekly (n = 169). The dose of darbepoetin alfa or epoetin was individually titrated to maintain hemoglobin concentrations within -1.0 to +1.5 g/dL (-10 to +15 g/L) of patients' baseline values and within a range of 9.0 to 13.0 g/dL (90 to 130 g/L) for up to 28 weeks (20-week dose-titration period followed by an 8-week evaluation period). The primary end point was change in hemoglobin level between baseline and the evaluation period (weeks 21 to 28). RESULTS: Mean changes in hemoglobin levels from baseline to the evaluation period were 0.24 +/- 0.10 (SE) g/dL (2.4 +/- 1.0 g/L) in the darbepoetin alfa group and 0.11 +/- 0.07 g/dL (1.1 +/- 0.7 g/L) in the epoetin group, a difference of 0.13 g/dL (95% confidence interval [CI], -0.08 +/- 0.33 [1.3 g/L; 95% CI, -0.8 to 3.3]). This difference was not statistically significant or clinically relevant despite the reduced frequency of darbepoetin alfa administration. The safety profile of darbepoetin alfa was similar to that of epoetin, and no antibody formation to either treatment was detected. CONCLUSION: These results show that darbepoetin alfa maintains hemoglobin concentrations as effectively and safely as epoetin in patients with CKD, but with a reduced dosing frequency. PMID- 12087569 TI - Once-weekly compared with three-times-weekly subcutaneous epoetin beta: results from a randomized, multicenter, therapeutic-equivalence study. AB - BACKGROUND: The ability to reduce the administration frequency of subcutaneous (SC) epoetin could provide benefits. This multicenter open-label study investigated the therapeutic equivalence of SC epoetin beta once-weekly and thrice-weekly administration regimens in maintaining anemia correction in stable hemodialysis (HD) patients. METHODS: One hundred seventy-three patients were randomly assigned to either once-weekly epoetin beta (n = 84) or their original thrice-weekly regimen (n = 89) for 24 weeks. All patients were administered intravenous iron supplementation, as required. RESULTS: The per-protocol analysis included 134 patients (69 patients, once-weekly group; 65 patients, thrice-weekly group). Mean hematocrits in both groups remained stable throughout the study. The difference in mean time-adjusted area under the curve for hematocrits between the once-weekly and thrice-weekly groups (-0.54 vol%) and 90% confidence intervals ( 1.27 to 0.19) were within the prespecified equivalence range (-2 to +2 vol%). There was no significant change in epoetin beta dose during the study. The ratio of mean weekly epoetin beta doses in the once-weekly and thrice-weekly groups (1.11) and 90% confidence interval (0.99 to 1.23) also remained within the prespecified range (0.8 to 1.25). Intention-to-treat analysis results were similar to per-protocol analysis results. Both regimens were well tolerated. CONCLUSION: Once-weekly and thrice-weekly SC epoetin beta administrations are statistically equivalent in terms of maintaining both stable hematocrits and epoetin beta dose requirements in HD patients. These findings may improve compliance among patients. PMID- 12087570 TI - Validity of subjective global assessment as a nutritional marker in end-stage renal disease. AB - Subjective global assessment (SGA) is a widely available clinical tool established to be of prognostic value in patients with end-stage renal disease (ESRD). Although it is reported to reflect a patient's nutritional status, its direct relationship to nutrition has not been determined. The aim of this study is to compare SGA with the gold standard for nutrition, total-body nitrogen (TBN) level, and thus determine the validity of SGA as a marker of nutritional status in patients with ESRD. Seventy-six consecutive dialysis patients referred from the renal service for routine measurement of TBN underwent simultaneous assessment of SGA determined by two independent examiners. Both examiners were blinded to TBN results. Only a moderate level of agreement was found in SGA score between the two examiners (weighted kappa score, 0.6). When patients were stratified into three nutritional groups determined by their SGA score, mean TBN values for each group by observer differed significantly (P = 0.0008 and P = 0.02, respectively). However, a significant statistical trend of worsening nutrition across SGA strata was found only for observer 2 (P = 0.049). Test performance of SGA as a predictor of malnutrition using a cutoff score of B (SGA = B or C) or C (SGA = C) was poor (positive likelihood ratios [LRs], 0.7 to 2.3; negative LRs, 0.5 to 1.0). Therefore, SGA appears not to improve the posttest probability of detecting malnutrition. SGA may differentiate severely malnourished patients from those with normal nutrition, but is not a reliable predictor of degree of malnutrition. PMID- 12087571 TI - Improvement of nutritional status after initiation of maintenance hemodialysis. AB - BACKGROUND: Recent data suggest that serum albumin concentrations increase during the several month-period that follows the initiation of maintenance hemodialysis (MHD) therapy. Some investigators have shown that the rate of increase in serum albumin level is related directly to 24-hour urine protein losses before the initiation of dialysis therapy. However, serum albumin levels increase even in patients starting MHD therapy without significant proteinuria, suggesting that this increase may be one manifestation of improving protein-energy nutritional status associated with commencing MHD therapy. We examined this question by reviewing records of all patients admitted to our outpatient dialysis unit within 30 days of initiation of MHD therapy. METHODS: Ninety-seven patients qualified for the study. Their routine laboratory test results beginning with the time of admission to the outpatient unit until month 6 of chronic dialysis therapy were analyzed. RESULTS: Between months 1 and 6, there was a statistically significant increase in values for predialysis serum albumin, iron, transferrin saturation, creatinine, and normalized protein equivalent of total nitrogen appearance (nPNA). Many of these values were still increasing by month 6. Body weight declined initially until month 4, then increased. There was a strong curvilinear relationship between mean nPNA and final serum albumin level. CONCLUSIONS: Our results suggest that an improvement in nutritional status, including an increase in dietary protein intake and serum albumin level, often occurs with the initiation of MHD therapy. The improvement in nutritional status may be related to the increase in dietary protein intake. PMID- 12087573 TI - An analysis of dialysis training in the United States and Canada. AB - BACKGROUND: Because the prevalence of end-stage renal disease (ESRD) has progressively increased in both the United States and Canada, patients with ESRD are likely to constitute progressively larger proportions of nephrology practices. METHODS: We mailed a questionnaire to US and Canadian nephrology program directors to determine methods used in dialysis training; 53% of US and 73% of Canadian programs responded. RESULTS: Training programs in the United States enrolled a larger median number of fellows and had a lower median faculty fellow ratio compared with programs in Canada. However, the availability of faculty in providing training in the care of patients undergoing maintenance hemodialysis (MHD) or chronic peritoneal dialysis (CPD) was similar in both countries. There were wide variations in availability of patients in both the United States and Canada. US training programs offered trainees significantly lower numbers of MHD and CPD patients; 29% of US training programs had less than five CPD patients per fellow. Similarly, there were wide variations in the amount of time trainees spent providing care to MHD and CPD patients; in 14% of US training programs, fellows spent less than 5% of their time receiving training for patients undergoing CPD. Only a small proportion of training programs had faculty resources or ensured training for fellows in the placement of percutaneous tunneled venous hemodialysis catheters or peritoneal dialysis catheters. CONCLUSIONS: To conclude, there are wide variations in dialysis training in both the United States and Canada. This survey raises concerns that many US training programs either do not have an appropriate number of CPD patients or do not allocate appropriate time to ensure the preparedness of fellows in providing independent care for patients with ESRD undergoing CPD. PMID- 12087572 TI - Improvement in nutritional parameters after initiation of chronic hemodialysis. AB - BACKGROUND: Protein-calorie malnutrition is highly prevalent in patients with chronic renal failure and on chronic dialysis therapy. Longitudinal studies evaluating nutritional outcomes after the initiation of chronic dialysis therapy in incident dialysis patients are limited. METHODS: This prospective cohort study evaluated time-dependent changes in several well-defined markers of nutritional status before and after initiation of chronic hemodialysis therapy. Fifty incident hemodialysis (HD) patients (60% men, 38% white, 32% with insulin dependent diabetes mellitus) were studied. Multiple nutritional markers, including biochemical parameters and analysis of body composition, were assessed before the initial outpatient CHD treatment and every 3 months thereafter for 12 months. RESULTS: At baseline, nutritional markers correlated well with each other. After the initiation of HD therapy, there were marked improvements in most nutritional parameters, including serum albumin, serum prealbumin, normalized protein catabolic rate, fat mass, reactance, and phase angle (P < 0.05 for all). Improvements in nutritional parameters were influenced by baseline nutritional status; ie, baseline nutritional parameters were predictors of their end-of-study value. CONCLUSION: Initiation of CHD therapy is associated with improvements in most nutritional markers. Nutritional benefits of increased solute clearance provided by the initiation of chronic dialysis therapy prevail over its potential catabolic effects. However, the extent of improvement was dependent on nutritional status at the time of initiation of dialysis therapy, which remained an important determinant of subsequent nutritional improvements during the first year of treatment. PMID- 12087574 TI - Outcome of peritoneal dialysis in cirrhotic patients with chronic renal failure. AB - BACKGROUND: Results of peritoneal dialysis (PD) in cirrhotic patients with renal failure are debated. The aim of the present study is to assess the outcome of PD patients with liver cirrhosis. METHODS: A retrospective study based on clinical records was performed in 21 cirrhotic and 41 PD patients without liver disease followed up at our PD unit. RESULTS: Five-year patient and technique survival were similar in the two groups. Seven cirrhotic patients (33%) died of liver failure (5 patients), liver cancer (1 patient), and peritonitis (1 patient). Ten controls (24%) died of cardiovascular complications (6 patients), pneumonia (1 patient), cachexia (1 patient), and unknown reasons (2 patients). Six cirrhotic patients discontinued PD therapy after 44.3 +/- 24.7 months because of inadequate dialysis (2 patients), sclerosing peritonitis (2 patients), dementia (1 patient), or patient choice (1 patient). Twelve controls discontinued PD therapy after 33 +/- 25.9 months because of peritonitis (6 patients), exit-site infection (1 patient), inadequate peritoneal clearance (3 patients), catheter malfunction (1 patient), and patient request (1 patient). The peritonitis rate was 0.31 episodes/y in cirrhotic patients and 0.53 episodes/y in controls (P = not significant). The hospitalization rate was similar (16.5 d/y in cirrhotic patients, 15.4 d/y in controls). In cirrhotic patients, complications were leakage (3 patients), symptomatic hypotension (5 patients), anemia (5 patients), severe malnutrition (14 patients), sclerosing peritonitis (2 patients), and hepatic encephalopathy (3 patients). CONCLUSION: The outcome of PD and risk for bacterial peritonitis or other dialysis complications are not worsened by the presence of liver cirrhosis. PMID- 12087575 TI - Genetic factors play an important role in the pathogenesis of hyperlipidemia post transplantation. AB - BACKGROUND: Our purpose was to identify factors associated with hyperlipidemia post-transplantation in a Hispanic population. METHODS: From 1985 to 1999, a kidney graft survival longer than 3 months occurred in 293 cases at the Instituto Nacional de la Nutricion. Most of the patients living in Mexico City were included (n = 83). The evaluation included a questionnaire, blood samples, and assessment of body composition and dietary habits. As many as possible first degree relatives were studied. RESULTS: Women had higher values of cholesterol (236 +/- 51 versus 215 +/-41; P < 0.05), low-density lipoprotein cholesterol (147 +/- 42 versus 131 +/- 34; P = 0.05), high-density lipoprotein cholesterol (57.3 +/- 14 versus 47.9 +/- 14; P = 0.002) and high-density lipoprotein-2 cholesterol. Isolated hypercholesterolemia was the most common lipid abnormality (40.9%), followed by mixed hyperlipidemia. Lipoprotein (a) greater than 30 mg/dL was found in 13 cases. Familial combined hyperlipidemia (FCHL) in the patient's relatives was a marker for dyslipidemia (odds ratio, 7.04; 95% confidence interval, 1.2 to 59.7). These cases had a worse lipid profile. Cyclosporine-treated FCHL patients had higher lipid levels compared with the non-FCHL, cyclosporine-treated patients. The effects of cyclosporine on the lipid levels were lower, but significant, after the exclusion of the FCHL cases. CONCLUSION: Post-transplant dyslipidemia is determined by genetic and environmental factors. FCHL in the patient's relatives was associated with post-transplant hyperlipidemia; an additive effect with cyclosporine was found. The evaluation of the lipid profile of relatives may be useful for the assessment of the risk of post-transplant dyslipidemia. PMID- 12087576 TI - Continued transplant immunosuppression may prolong survival after return to peritoneal dialysis: results of a decision analysis. AB - BACKGROUND: Patient survival is prolonged in those with residual renal function. The risks of opportunistic infection and malignancy militate against a widespread policy of maintaining immunosuppression in those returning to peritoneal dialysis after a failed allograft. We set out to test the hypothesis that patients returning to peritoneal dialysis after a failed transplant benefit from continued immunosuppression. METHOD: A decision analytic model comparing the use of immunosuppression after transplant failure and return to peritoneal dialysis with immunosuppressive withdrawal. Two assumptions were made: that the survival benefit reported in patients with a transplant kidney was the same as that expected from a native kidney with a similar glomerular filtration rate (GFR) and that the risks of carcinoma and opportunistic infections were equal to that of the general population if immunosuppressive therapy was discontinued. Probabilities were obtained from the literature, from an administrative database, and from the rate of GFR decline seen in our own unit in patients returning to dialysis who had been discontinued from their transplant immunosuppressive medication. RESULTS: Life expectancy was prolonged from 5.3 years to 5.8 years when immunosuppression was continued in patients who returned to dialysis after chronic allograft failure. A higher survival benefit was seen at higher levels of additional GFR. A benefit from continued immunosuppressive therapy was seen for all values of additional GFR greater than 15 L/wk. All dialysis and renal clearances have been expressed as L/wk. The conversion factors for conversion to mL/s and mL/min are 605 and 10.1. CONCLUSION: The data suggest that there may be a survival advantage in maintaining patients on long-term immunosuppressive function even after they return to peritoneal dialysis after allograft failure. PMID- 12087577 TI - Can glomerular mRNAs in human type 1 diabetes be used to predict transition from normoalbuminuria to microalbuminuria? AB - BACKGROUND: mRNAs of pathogenetic importance in the development of diabetic nephropathy were measured in subjects with type 1 diabetes to determine whether these might be used to predict progression from normoalbuminuria to microalbuminuria. We proposed that conversion from normoalbuminuria to microalbuminuria would be most likely in subjects whose connective tissue growth factor (CTGF) and collagen mRNAs were above the 95% confidence interval (CI) for live renal donors and within the 95% CI for subjects with abnormal albuminuria. METHODS: Glomerular CTGF, collagen alpha2(IV), and control glyceraldehyde-3 phosphate dehydrogenase (GAPDH) mRNAs were measured in microdissected glomeruli from living renal donors (n = 10), and subjects with normoalbuminuria (n = 12), microalbuminuria (n = 5), and overt proteinuria (n = 6). RESULTS: After 44 +/- 2 months of follow-up, one subject converted from normoalbuminuria to microalbuminuria. Although the data are limited, progression from normoalbuminuria to microalbuminuria occurred in the only normoalbuminuric subject whose mRNA levels were above the live renal donors' 95% CI for CTGF and collagen alpha2(IV) and within the 95% CI of subjects with abnormal albuminuria. No clinical or histopathologic finding distinguished the progressor from the nonprogressors at the time of biopsy. CONCLUSION: This case report provides proof of-principle that a panel of glomerular mRNA markers chosen because of their pathogenetic relevance may be useful adjuncts to albuminuria and histology in predicting clinical stability or clinical progression in diabetic nephropathy. PMID- 12087578 TI - Treatment of acute renal failure caused by renal artery occlusion with renal artery angioplasty. AB - BACKGROUND: Renovascular disease is a common cause of renal impairment and hypertension, particularly in the older population. Oligoanuric acute renal failure secondary to renal artery occlusion is not well recognized; however, it is potentially reversible if identified and treated. METHODS: Five patients presented to our institution with oligoanuric acute renal failure. Each had evidence of vascular disease, and a prerenal insult was identified. They were investigated with renal artery Doppler ultrasound or nuclear imaging before proceeding to percutaneous angioplasty and stent placement. RESULTS: The targeted kidney had relatively well-preserved renal size, and potential viability of the renal tissue was determined by nuclear scanning with parenchymal uptake of tracer. Percutaneous angioplasty and stent placement resulted in brisk reperfusion of the kidney and an immediate diuresis with improvement of renal function, avoiding supportive dialysis after the procedure. Contrast nephrotoxicity was identified in two of the five cases. CONCLUSION: Renal artery occlusion should be considered as a cause of oliguric acute renal failure, particularly in patients at high risk who present with a sudden deterioration of renal function, with nuclear imaging showing potentially viable renal tissue with relatively well-preserved renal size. Percutaneous revascularization should be considered in this group. PMID- 12087579 TI - A late episode of post-transplant diabetes mellitus during active hepatitis C infection in a renal allograft recipient using tacrolimus. AB - BACKGROUND: An association between hepatitis C virus and (post-transplant) diabetes mellitus has been reported. METHODS: We report a patient on tacrolimus based immunosuppression who developed an episode of post-transplant diabetes mellitus (PTDM) 2 years after renal transplantation, after contracting a hepatitis C infection. Her glucose metabolism was evaluated regularly by intravenous glucose tolerance tests before and after the PTDM episode. RESULTS: Before contracting hepatitis C, the patient's insulin resistance and insulin secretion were normal. After contracting hepatitis C, tacrolimus exposure increased, insulin resistance increased, and insulin secretion decreased markedly. Despite low tacrolimus exposure in the last 4 years, glucose metabolism did not recover completely. Although PTDM resolved and insulin resistance normalized, pancreatic beta cell secretion remained impaired by approximately 50% compared with the period before hepatitis C infection. CONCLUSION: After an initial increase in insulin resistance, insulin secretion decreased markedly in a patient who contracted hepatitis C 12 to 22 months after renal transplantation. This change resulted in an episode of PTDM. Increased tacrolimus exposure secondary to reduced cytochrome P-450 metabolism as a result of impaired hepatocellular function at the time of the development of PTDM seems a likely explanation for the marked decrease in insulin secretion. Viral toxicity to the beta cell might be an additional explanation. The latter might be suspected from several reports about an association between diabetes mellitus and hepatitis C in patients who do not use drugs that interfere with glucose metabolism. PMID- 12087580 TI - Lead, renal disease and hypertension. PMID- 12087581 TI - The nutritional status of chronic renal failure patients following the initiation of hemodialysis treatment. PMID- 12087582 TI - Dialysis training in the United States. PMID- 12087583 TI - A 50-year-old man with persistent renal allograft dysfunction. PMID- 12087584 TI - Chinese herbs nephropathy: not merely a trivial matter of a name. PMID- 12087585 TI - Lunch-related polyuria. PMID- 12087586 TI - Parenteral iron safety and the special article "Just the FAQs: frequently asked questions about iron and anemia in patients with chronic kidney disease". PMID- 12087588 TI - Pamidronate-associated nephrotoxicity in a patient with Langerhans's histiocytosis. AB - Pamidronate is used frequently to treat malignancy-associated hypercalcemia and osteolytic lesions. This widely used bisphosphonate has been noted to cause nephrotoxicity in patients with multiple myeloma and metastatic breast cancer. We encountered a patient with Langerhans's histiocytosis who developed nephrotic syndrome and renal failure after pamidronate therapy. We describe the clinical and renal biopsy findings in this patient. PMID- 12087587 TI - Familial membranoproliferative glomerulonephritis type III. AB - BACKGROUND: Membranoproliferative glomerulonephritis (MPGN) is a relatively uncommon cause of progressive renal disease characterized by immune complex deposition resulting in mesangial proliferation and endocapillary inflammation with capillary wall thickening and double contour formation. Although a familial linkage has been reported in MPGN type II disease and less often in type I disease, a familial linkage in type III disease has not been reported previously. METHODS: We identified a family in which MPGN type III developed in a living related donor 12 years later and recurred in the renal allograft of his son, whose primary disease was MPGN type III. We screened the members of the extended family, looking for evidence of hematuria and proteinuria. Renal biopsy specimens exhibited the findings of subendothelial deposits, subepithelial deposits, and complex glomerular basement membrane changes with C3 but not IgG seen on immunofluorescence. RESULTS: Screening identified eight affected family members (six biopsy proven) over three generations. The condition is inherited in an apparent autosomal dominant fashion. CONCLUSION: This is the first description of familial MPGN type III. We hope that by studying the disease in this family group, we may learn more about the pathogenesis of the condition. PMID- 12087589 TI - Enteric hyperoxaluria: a hidden cause of early renal graft failure in two successive transplants: spontaneous late graft recovery. AB - A 37-year-old patient underwent two successive renal transplantations 7 months apart. He remained dialysis dependent. Early biopsy of both grafts revealed widespread calcium oxalate deposition suggestive of acute oxalate nephropathy. Several causes of oxalate nephropathy, including primary oxalosis and an increased intake of oxalic acid precursors, were excluded. Two years later, the identification of steatorrhea with radiologic signs of chronic pancreatitis led to the hypothesis of enteric hyperoxaluria. Surprisingly, 11 months after the second transplantation, graft function improved progressively allowing interruption of dialysis. Three years later, renal function is stable. The causes and prevention of acute oxalate-induced graft failure are highlighted. Subclinical evidence of enteric hyperoxaluria should be looked for and appropriate therapy instituted as early as possible. The possibility of a late recovery of renal function warrants attentive patience from attending physicians. PMID- 12087590 TI - Gaucher disease with nephrotic syndrome: response to enzyme replacement therapy. AB - Nephrotic syndrome in patients with Gaucher disease is rare; most of the few reported cases have had a well-defined glomerulopathy often with Gaucher cells in the glomeruli. We report the case of a 54-year-old woman with Gaucher disease, who had splenectomy at age 25, preeclampsia with renal biopsy disclosing only endotheliosis at age 32, and improvement of proteinuria and reappearance of heavy proteinuria (7.2 g/24 h) at age 41. Renal biopsy disclosed Gaucher cells in glomeruli and interstitium. The patient did not receive therapy specifically for glomerular disease. Enzyme replacement, begun 4 years later and maintained until now, was associated with amelioration of systemic symptoms and virtual disappearance of proteinuria with a follow-up of 10 years. This case apparently is the first instance of nephrotic syndrome consequent to Gaucher disease itself and successful treatment with specific enzyme replacement. PMID- 12087591 TI - Viewpoint: hospitals often play important role as intermediaries of patients in clinical research programs. PMID- 12087592 TI - Just how important is book-larnin' at the bedside anyway? PMID- 12087593 TI - The A-B-C's of the nursing crisis. PMID- 12087595 TI - Spur staff to better document patient education. PMID- 12087594 TI - Simplicity is the best medicine for compliance information. Eight basic steps help improve employee comprehension. PMID- 12087596 TI - Food allergies--keeping kids safe. PMID- 12087597 TI - Herpes: a bigger problem than you think. PMID- 12087598 TI - The emergency management plan. PMID- 12087599 TI - A lesson in poor care. PMID- 12087600 TI - Angioplasty & stenting in the carotid? PMID- 12087601 TI - Surgical tape. PMID- 12087602 TI - Keep electronic records safe! PMID- 12087603 TI - Treatment costs of venlafaxine and selective serotonin-reuptake inhibitors for depression and anxiety. AB - In this article, health care expenditures are assessed for patients diagnosed with depression who are being treated with either venlafaxine (immediate or extended release) or a selective serotonin-reuptake inhibitor (SSRI). Patients beginning treatment for a new depressive episode were identified retrospectively from 1994 to 1998. Before beginning therapy, patients prescribed venlafaxine (N = 353) had more nonmental illnesses (0.84 vs. 0.75 clinical events/patient, respectively; P < .01) and hospitalizations for mental illness (0.56 vs. 0.30 hospitalizations/patient; P = .06) than patients prescribed SSRIs (N = 7,330). In the six months after initiating treatment, venlafaxine was associated with lower hospitalization expenditures for nonmental illness than were SSRIs ($206 vs. $472, respectively; P = .02), but total health care expenditures were not significantly different. PMID- 12087604 TI - Managed care implications of the approval of generic or over-the-counter nonsedating antihistamines. PMID- 12087606 TI - Disease management or population health? An example for program development. PMID- 12087605 TI - Escherichia coli resistance in uncomplicated urinary tract infection: a model for determining when to change first-line empirical antibiotic choice. AB - Escherichia coli is typically the causative organism in uncomplicated urinary tract infection (UTI). Resistance rates of E. coli to trimethoprim/sulfamethoxazole (TMP/SMX) are increasing, exceeding 10% in many communities. Guidelines recommend using alternative treatments in these areas. Providers must reevaluate policies to include considerations for E. coli resistance. A model was developed, with cases for illustration, to help organizations determine the resistance rate threshold, where TMP/SMX is no longer first-line therapy. Using published data, a 19% to 21% threshold was derived, supporting a previous report of 22%. The model can aid decision makers updating internal policies to conform with guidelines for the treatment of uncomplicated UTI and to improve care. PMID- 12087607 TI - Achieving optimal maternal and infant health outcomes for Medicaid patients, with application for commercial populations. AB - Maternity management, the cornerstone of Medicaid programs, will support the nation's ability to achieve public health goals related to maternal and infant health. The authors' present data showing how organizations that combine practical solutions garnered from both public and commercially funded programs have the greatest potential for developing successful maternity management programs. PMID- 12087608 TI - Bridging the gap: state and private industry response to the lack of a Medicare outpatient prescription drug benefit. AB - Although there is considerable political momentum and public support for a Medicare outpatient prescription drug benefit, enactment of full-fledged reform will take time given the other political priorities on the legislative agenda. The article describes how various states and private initiatives have acted to increase access to affordable outpatient prescription drugs for seniors. PMID- 12087609 TI - Time for physician groups to lead the way toward an electronic medical record. PMID- 12087610 TI - Stats & facts. Centers of excellence and managed care. PMID- 12087612 TI - Audio for bio-communicators. PMID- 12087613 TI - Digital biomedical. Photojournalism. AB - This article describes the strategies used to successfully complete a digitally based biomedical photojournalism assignment. A multi-step approach is suggested which includes project and funding identification, photographic planning, on-site photography and post project follow-up. Practical suggestions for utilizing digital imaging are included. PMID- 12087611 TI - Lymph node. Evaluation. AB - The photographs represent a novel clinical approach to the identification of sentinel lymph nodes (SLN). They are unique in the photographer's Canadian institution as they represent the first example of digital photography where time was limited by the nature of the medical procedure. The photographs were shot using an Olympus C2500 at 1712 x 1368 pixels using the super macro feature of the camera, manually white balanced to the operating room lights. PMID- 12087614 TI - A look at the problem of falls among people with cancer. AB - People with cancer often have multiple risk factors for falls. A major risk factor is aging; because people often are diagnosed with cancer at an older age or are living longer with cancer, many are at risk for falls. Additional risk factors include impaired physical function, sensory-neurologic deficits, use of multiple medications, and deconditioning, often caused by treatment-induced fatigue. Of all types of accidents, falls pose the most serious threat to the elderly. Fall injuries can have serious consequences related to reduced physical functioning and quality of life. A detailed understanding and awareness of the risk factors (both intrinsic and extrinsic) and problems associated with falls can assist oncology nurses to be vigilant in the identification of patients at high risk for falls. These patients may benefit from protocols used in other patient populations, new interventions, or the use of assistive devices. PMID- 12087615 TI - Granulocyte macrophage colony-stimulating factor: current practice and novel approaches. AB - Endogenous myeloid colony-stimulating factors (CSFs) have demonstrated the ability to enhance the clinical management of immunosuppressed patients with cancer. These agents are associated with significant decreases in chemotherapy associated infections, antibiotic use, length of hospital stays, and mortality. Two major endogenous recombinant myeloid CSFs currently are being manufactured. Granulocyte macrophage CSF (GM-CSF) (sargramostim, Leukine, Immunex Corporation, Seattle, WA) has broad activity in the proliferation and differentiation of myeloid lineage progenitor cells, whereas granulocyte CSF (filgrastim, Neupogen, Amgen, Inc., Thousand Oaks, CA) acts selectively on cells of the granulocyte lineage. Clinical trials suggest that GM-CSF has clinical benefits beyond enhancing neutrophil recovery, including shortening the duration of mucositis and diarrhea, stimulating dendritic cells, preventing infection, acting as an adjuvant vaccine agent, and facilitating antitumor activity. PMID- 12087616 TI - Effects of cancer-related anemia on clinical and quality-of-life outcomes. AB - Significant progress has been made in the prevention and management of many symptoms associated with cancer and its therapy. However, cancer-related anemia, which may affect more than half of all patients with cancer, often is assessed and treated inadequately. Severe anemia not only causes significant symptomology but also may impair functional status and quality of life. In addition, anemia has been found to affect treatment outcomes, including disease-free and overall survival. Therapeutic strategies, based on the underlying etiology of anemia, include iron supplementation, blood transfusion, and administration of recombinant human erythropoietin. Future approaches may involve novel agents under investigation that are designed to address current concerns related to efficacy, convenience, and possible cost-effectiveness of treatment of cancer related anemia. PMID- 12087617 TI - Consequences of chemotherapy on the sexuality of patients with lung cancer. AB - Chemotherapy can cause a variety of physical and emotional changes that affect all aspects of patients' lives, including sexuality. Alterations in physical appearance can significantly influence people's perceptions of their sexual identities, attractiveness, and worthiness. Patients with lung cancer receiving chemotherapy may need sexual counseling. Therefore, patients and healthcare providers should create an environment that allows adequate time to discreetly discuss the impact that chemotherapy treatment may have on appearance, self esteem, and sexuality. Nurses and physicians might hesitate to discuss sexuality with patients for various reasons, including time limitations, privacy considerations, readiness, and comfort level. Employers should provide support, educational programs, and professional resources so that healthcare providers can obtain the knowledge, develop the skills, and recognize that counseling patients about sexual issues is an important aspect in providing comprehensive holistic care to patients with lung cancer. PMID- 12087618 TI - Providing information to patients with a rare cancer: using Internet discussion forums to address the needs of patients with medullary thyroid carcinoma. AB - Medullary thyroid carcinoma (MTC) is a rare disease, and specific experience with the condition is not commonly found outside of major cancer centers. Because of the rarity and slow-growing natural history of the disease, patients with MTC frequently have many questions concerning prognosis, treatment, and follow-up care. Many of these patients have joined an e-mail group on the Internet through which they share information and concerns. The author has joined this group and shares her expert clinical knowledge about the disease. When individuals are armed with accurate and expert information, they can make informed decisions. The source of this information can be diverse. Nurses with expert knowledge can use opportunities such as e-mail groups to answer questions and respond openly about subjects about which they have unique perspectives. PMID- 12087619 TI - Putting your best foot forward in a challenging role: finding the resources needed to work in a freestanding radiation oncology clinic. AB - This article describes the experiences of a nurse in a new role in a freestanding radiation oncology clinic. Networking to find the resources that patients need and providing guidance to the patients in using the resources through their course of treatment are discussed. Local and national resources that can be used as tools in radiation therapy nursing also are described. PMID- 12087620 TI - To draw or not to draw: drawing blood cultures from a potentially infected port site. PMID- 12087621 TI - Coping with cancer resource list. PMID- 12087622 TI - Cachexia in patients with advanced cancer. AB - Cancer cachexia generally is considered to be the end stage in the progression of nutritional deterioration and wasting of malignancy (Ottery, 1995). In patients with advanced cancer, this condition is very common and decreases quality of life, as well as survival (Fearon et al., 2001; Ottery; Smith & Souba, 2001; Whitman, 2000). However, if early diagnosis and intervention can control cachexia, the potential exists to greatly improve a patient's quality of life and prolong survival. Because metabolic alterations inhibit the effective use of conventional nutritional support, anti-inflammatory agents or fish oil are possible options. Orexigenic agents may be prescribed if patients wish to improve oral intake. Steroids and progestational agents may be used to attempt to improve mood and appetite. Nutrition affects symptoms that need to be managed effectively. Nurses should work aggressively to correct factors that contribute to decreased food intake (e.g., nausea, pain) and correct factors that worsen debility (e.g., anemia). Information must be presented so that informed choices can be made and realistic eating goals set. An interdisciplinary approach that involves the nurse, physician, dietician, and possibly social worker or case manager, as well as the patient and family, is necessary to identify nutritional alterations, assess specific needs, and plan individual interventions. Whitman (2000) stated that counseling is the most effective and least expensive intervention. It may be conducted by any member of the healthcare team and should be combined with other interventions. Palliation of cachexia in patients with advanced cancer is a challenge for nurses. Hopefully, early and judicious use of these interventions may decrease the significant morbidity and mortality that result from cancer cachexia. PMID- 12087623 TI - Psychoneuroimmunology. Part II: Mind-body interventions. AB - PNI provides the scientific foundation of several integrative therapies. PNI interventions or mind-body therapies can be categorized into four general mechanisms: sensory, cognitive, expressive, and physical. Some therapies are used in combination, and others incorporate more than one mechanism. Although PNI is limited as a purely biomedical framework, it is a valuable frame of reference because it supports the use of many integrative therapies for patients with cancer. PMID- 12087624 TI - Epirubicin hydrochloride. PMID- 12087625 TI - WHO study of venous thrombosis and air travel. PMID- 12087626 TI - Status incongruity in Samoan youth: a biocultural analysis of culture change, stress, and immune function. AB - In Samoa, the presence of a matai title in the family has historically been a valued source of social status. However, as the process of Westernization continues, new sources of social status are emerging. This study explores the degree to which new and old markers of social status agree--or disagree--and the consequences they have for the experience of stress in 329 Samoan adolescents. The study integrates cultural and biological methods and data, and measures an aspect of immune function (antibodies against the Epstein-Barr virus) as a biomarker of psychosocial stress. Results indicate that status "incongruent" adolescents experience significantly more stress (indicated by reduced immune function), and that emerging markers of social status are becoming inextricably linked to "traditional" markers in such a way that discordance between them is a significant source of stress. This study proposes new conceptual models for future studies of culture change and suggests that biomarkers may represent ethnographic tools that can provide insight into hidden cultural dynamics and the experience of stress. PMID- 12087628 TI - African independent churches in Mozambique: healing the afflictions of inequality. AB - The recent explosive proliferation of African Independent Churches (AICs) in central Mozambique coincided with rapid growth of economic disparity in the 1990s produced by privatization, cuts in government services, and arrival of foreign aid promoted by Mozambique's World Bank/International Monetary Fund Structural Adjustment Program. Drawing on ethnographic research in the city of Chimoio, this article argues that growing inequality has led to declining social cohesion, heightened individual competition, fear of interpersonal violence, and intensified conflict between spouses in poor families. This perilous social environment finds expression in heightened fears of witchcraft, sorcery, and avenging spirits, which are often blamed in Shona ideology for reproductive health problems. Many women with sick children or suffering from infertility turn to AICs for treatment because traditional healers are increasingly viewed as dangerous and too expensive. The AICs invoke the "Holy Spirit" to exercise malevolent agents and then provide a community of mutual aid and ongoing protection against spirit threats. PMID- 12087627 TI - Giving birth to gonolia: "culture" and sexually transmitted disease among the Huli of Papua New Guinea. AB - The "culture concept" has been challenged on a number of fronts, both by medical anthropologists researching AIDS and in the discipline of cultural anthropology more generally. Medical anthropologists have argued against the "etiologization" of culture, and cultural anthropologists have taken issue with the tendency to treat beliefs and practices as static and seamlessly shared. Using the narrative of one Huli woman's shifting explanation of a diagnosis of syphilis, this article argues that, rather than avoid the notion of culture, we should strive for representations that demonstrate how individuals use discourses in expedient, ad hoc, and yet deeply felt ways. This article also argues for the importance of a sociology of knowledge approach to understanding local notions of etiology. The woman's understanding of her situation was strongly influenced by her entry into a new "community" of women who had similarly been diagnosed with a sexually transmitted disease. PMID- 12087629 TI - Three days for weeping: dreams, emotions, and death in the Peruvian Amazon. AB - Funeral customs and beliefs about the dead among the Matsigenka of Peru reflect great anxiety over the contagion of death and the power of the dead over the living. In this article, these beliefs and practices are interpreted in light of a native theory about emotion. For the Matsigenka, excessive displays of emotion, especially grief, anger, and aggressive sexuality, cause a vicious cycle of unhappiness, social disruption, illness, and death. Moving beyond a culturally particular, symbolic-interpretive perspective, the author also draws on personal experiences both in the field and at home to reflect on the phenomenology of grief across cultures. PMID- 12087630 TI - Disparate views of community in primary health care: understanding how perceptions influence success. AB - The importance of community in primary health care (PHC) is evident in the role of community participation and in the types of programs that are routinely implemented (community health-worker [CHW] programs, community clinics, community based disease-control programs). Few health care providers and program administrators, however, have considered the meaning of community. Instead, they frequently impose their own definition of community and assume that it corresponds to local realities. This is problematic because target populations may have different ideas about what a community is and how it functions. When disparate ideas of community exist, they can affect the implementation of PHC programs, leading to low rates of acceptance, participation, and utilization. Using two examples, a community clinic in El Alto, Bolivia, and a CHW program in Rio Branco, Brazil, this article discusses some of the problems that arise when local definitions of community do not correspond to programmatic definitions. PMID- 12087631 TI - NJSNA members participate in multi-state study to assess safe patient handling. PMID- 12087632 TI - Evolving toxicities associated with anti-HIV therapies. PMID- 12087633 TI - Harm reduction psychotherapy: an innovative alternative to classical addictions theory. AB - Harm Reduction Psychotherapy is an innovative treatment for people with alcohol and other drug problems. Unlike the traditional disease model of addiction, HRP uses a biopsychosocial approach to understand the complexities of drug use, abuse, and addiction. In other words, in the context of HRP, addiction is not the primary issue. Rather, it is an interactive phenomenon in which the relative weight of biology, psychology, and social factors varies for each person and for each drug he or she uses. HRP allows us to assess each person individually and to plan treatment that is tailored to the individual's relationship with alcohol and other drugs. It also incorporates other important problems: emotional disorders, family problems, social alienation, and medical complications. These issues are discussed at the beginning of consultation, without patients having to focus solely on their alcohol or drug problem. The unique aspect to HRP is that patients do not have to commit to abstinence as a condition of, or even necessarily as a goal of, treatment. HRP seeks to identify and work with the barriers to treatment adherence in any patient. It is clear that most medical patients have some difficulty understanding and adhering to medical recommendations and treatment protocols. However, drug users have particular problems that must be identified. HRP helps people create individual strategies to decrease harmful alcohol and drug use. It uses a nonjudgmental and collaborative approach to actively encourage individuals to explore their own barriers to change and to choose among a range of options such as abstinence, moderation, or other short-term goals. Motivational interviewing can be used to motivate behavioral change with the goal of reducing the effects of adverse consequences. PMID- 12087634 TI - The use of alternative medicine in the treatment of hepatitis C. AB - More than one-third of Americans use herbs for health purposes, yet patients and physicians usually lack accurate information about safety and efficacy of herbal remedies. In recent years, there has been a substantial increase in the use of so called complementary and alternative therapies by patients with liver disease. Medical professionals and laboratorians need to be informed about popular alternative therapies and be open-minded to the possibility that some benefit may come from some therapies currently regarded as alternative. Silymarin extracted from the milk thistle is most widely subscribed to as a remedy for liver diseases. The beneficial effects of silymarin are most often seen in the patients who had cirrhosis as a result of alcohol abuse. An ongoing clinical trial will provide some insight as to whether milk thistle directly affects HCV. Silymarin has a good safety record and only rare case reports of gastrointestinal disturbances and allergic skin rashes have been published. The active component of licorice root, glycyrrhizin, has been shown to reduce alanine transaminase and aspartate transaminase values in the serum. This protective function has recently been explained as the inhibitory effects of glycyrrhizin on immune-mediated cytotoxicity against hepatocytes and on nuclear factor (NF)-kappa B, which activates genes encoding inflammatory cytokines in the liver. Finally, some patients with hepatitis C take St. John's Wort and ginger to treat the side effects caused by interferon therapy. An excellent review of this subject was recently published by the NCCAM. PMID- 12087635 TI - A role for the novel weight-regulating hormone ghrelin in anorexia nervosa. PMID- 12087636 TI - Comparative evaluation of serum protein analysis by capillary zone electrophoresis and agarose gel electrophoresis. PMID- 12087637 TI - Quantitative estimation of organic analytes with a capillary column. PMID- 12087638 TI - LabAutomation 2002: productive technologies for the new millennium distributed laboratories and point-of-care--Part 2. PMID- 12087639 TI - Current approaches to classic HIV paradigms. PMID- 12087640 TI - A service-oriented teaching and learning project. AB - Profound changes in the health care system have heightened nursing educators' awareness about preparing students for the role of health educator to patients and families. To address this concern, senior students in a child and family course developed a teaching and learning (TL) project oriented toward children's and families' distinctive learning needs. With the clinical instructor's guidance, the students sought to create a unique learning activity while still meeting the course's TL objectives. Students gathered information on an array of health topics and determined that a "mini-health fair" for children, families, and professionals using attractive and informative posters and presentations was an effective strategy for disseminating the information. Participants listened to the students' presentations and viewed posters. Pediatric patients enjoyed the interactive learning, families appreciated the take-home informational handouts, and clinicians applauded the professionalism of the student's work. The TL project that was designed to develop students' skills as educators developed into a countywide experience. Students participated in the community's annual health fair and presented to the nursing school's governing board as an exemplar of practice-based teaching. Students were successful when given encouragement and support to create and present TL projects. PMID- 12087641 TI - A school-based intervention to teach third grade children about the prevention of heart disease. AB - Coronary heart disease continues to be a leading cause of death in adults. Because many risk factors for heart disease make their first appearance in childhood, early intervention is an attractive strategy for this disease. A convenience sample of 58 third grade children participated in a year-long, quasi experimental study to compare a school-based educational program, HeartPower!, to the standard health education curriculum. Variables examined included knowledge, hypercholesterolemia, obesity, sedentary lifestyle, hypertension, exposure to cigarette smoke, and parental participation. The students who participated in the HeartPower! Program showed improvement (p < 0.05) in their knowledge of healthy lifestyles. Modest improvements in hypertension and exposure to tobacco smoke were observed. These results indicate that focused instruction on important health issues may be preferable to a broad approach. PMID- 12087642 TI - The pediatric nurse's role as health consultant to a child care center. AB - An experienced pediatric nurse can perform a valuable service by becoming a health consultant to a child care center. Consultation and collaboration with child care providers and parents can foster health and safety for the steadily increasing number of young children being cared for in this type of community setting. The nurse with a strong pediatric background possesses the expert knowledge and skills needed to promote health and prevent illness and injury in this population. To exemplify the typical tasks encountered by a child care health consultant, a case is described in which a daycare director contracts the services of a nurse consultant to address problems with infection control procedures and illness transmission at a particular center. The consultant organizes the approach to this assignment by using a nursing consultation model as an action guide to achieve a positive outcome for the consultee. PMID- 12087643 TI - Raising the bar for health and safety in child care. AB - During the past few decades, this country has experienced many socio-economic changes including a rise in the number of dual-income families and single parent homes. Combined with a fluctuating economy and drastic reforms in welfare, one of the results has been an ever-increasing number of children being cared for in out of-home settings. With almost 75% of all children under age 5 years and 50% of infants in some form of child care on a regular basis, opportunities abound for nurses to promote optimal health and safety in child care. To ensure quality child care for these children, health care professionals are uniquely positioned to provide consultation services to centers and family child care settings. With expertise in child development, infection control, disease prevention, and health promotion, pediatric and public health nurses can provide many types of child care health consultation services to impact the care of these children positively. PMID- 12087644 TI - "Dude, you don't have Tourette's:" Tourette's syndrome, beyond the tics. AB - While coprolalia is the most well-known symptom of Tourette's syndrome (TS), it affects only a minority of persons with the condition. TS is a chronic neurobiological condition consisting of vocal and motor tics. Many children with TS have associated obsessive-compulsive disorder (OCD) and/or attention deficit hyperactivity disorder (ADHD), both of which can interfere with school, peer, and family functioning more than the tics themselves. The article distinguishes TS from other tic disorders and reviews literature on epidemiology, etiology, clinical course, and diagnostic and treatment issues. The article discusses the role of primary care pediatric and advanced practice nurses in the diagnosis and management of TS and details helpful interventions in the arenas of personal, family, and educational support, as well as symptom management and indications for medications. The author also suggests areas for future nursing research. PMID- 12087645 TI - What do parents of children with learning disabilities, ADHD, and related disorders deal with? AB - Children with learning disabilities, attention deficit/hyperactivity disorder (ADHD), and related disorders puzzle parents because of their many abilities and disabilities. It can also be difficult to understand how much of their behavior is the nature of the condition and how much is oppositional. It is all too easy for parents to "catch" a child's feelings of inadequacy and then feel bad as a parent. Parenting approaches that include clear, concise instructions; structure without rigidity; nurturing a child's gifts and interests; and constant approval of positive behavior help parents feel better and help children feel safe. It takes time for both children and parents to embrace the concept that being different does not mean being inferior and, in fact, can be a good thing. Parents need to be nurtured and praised to help them nurture and praise their children. Most parents use almost every resource they have to help their children flourish, and still, they worry they are not doing a good enough job. Usually they are! PMID- 12087646 TI - Altered level of consciousness in a 2-year-old. PMID- 12087647 TI - Key steps in implementing evidence-based practice: asking compelling, searchable questions and searching for the best evidence. AB - From the searches presented, it can be seen that there are times when scientific studies will not be available to answer burning clinical questions. If systematic reviews, evidence-based guidelines, and individual studies are not available, clinicians must then turn to expert anecdotal evidence to guide their decision making process and interventions. As further randomized controlled trials are conducted to evaluate nursing interventions, more scientific evidence will become available to guide nursing practice. However, it is important to remember that EBP not only encompasses evidence from research and its appraisal, but also includes patient preferences and expertise by the clinician. Because a meta analysis indicated that patients had 28% better outcomes when nursing interventions were based on scientific evidence rather than interventions that were steeped in tradition (Heater, Becker, & Olson, 1988), it is essential that nurse practitioners and nurses have the appropriate knowledge and skills to translate evidence from research into practice. Asking searchable questions and knowing how to search for and find the best evidence are two key steps in delivering the highest quality of evidence-based nursing care to patients and their families. PMID- 12087648 TI - Update on newborn hearing screening programs. AB - As a result of legislation supporting the position statement of the American Academy of Pediatrics (AAP) Joint Committee on Hearing, newborn hearing screening programs have become a protocol of care in hospital nurseries throughout the United States. The current goals are to screen all infants by 1 month of age, confirm hearing loss with audiologic examination by 3 months of age, and treat with comprehensive early intervention services before 6 months of age. This article will review the development of the Joint Committee on Infant Hearing position statements, examine legislation and health policy initiatives for universal newborn hearing screening, and determine nursing's role in meeting the goals. PMID- 12087649 TI - A sugar-coated pacifier reduces procedural pain in newborns. AB - PURPOSE: To examine the efficacy of pacifiers and sugar, alone and in combination, for pain management in neonates. METHODS: An experimental design examined pain responses of 84 newborns undergoing heelstick. They were randomly assigned to one of four groups: (a) water-moistened pacifier, (b) sugar-coated pacifier, (c) 2 cc of a 12% oral sucrose solution, or (d) control. Pain measures were duration of cry, vagal tone, and salivary cortisol. RESULTS: Multivariate analysis of variance (MANOVA) revealed that the sugar-coated pacifier group cried significantly less than the water-moistened pacifier and control groups. Repeated measures analysis of variance (ANOVA) revealed that the sugar-coated pacifier group demonstrated significantly lower vagal tone during heelstick than the oral sucrose solution and control groups. This difference between the sugar-coated pacifier and control groups persevered for 15 minutes after heelstick. CONCLUSIONS: Offering a sugar coated pacifier during heelstick in healthy neonates reduces pain behaviors more effectively than a water-moistened pacifier, 2 cc of a 12% sucrose solution, or no intervention. PMID- 12087650 TI - Failure of mental health policy--incarcerated children and adolescents. AB - The difficulty surrounding mental health treatment of juvenile offenders is often a struggle to balance the overall social need for security with the needs of the individual. Rather than recognize the mental health care needs of youth in the juvenile justice system, the current health care environment will often shift responsibility and service costs for the most difficult to manage clients, creating a prison system that acts as a mental hospital. This article will discuss the disparities in accessing treatment for juveniles and review key legislation that addresses child mental health needs. PMID- 12087651 TI - A new profile of the knowledge, skills, and abilities of the pediatric nurse: a revision of the CPN certification exam. AB - Recently an expert panel of pediatric nurses from the U.S. and Canada, using data gathered from interviews, activity logs, and aspects of an international role delineation survey, created a new national pediatric nursing exam content outline that reflects the nature of general pediatric nursing practice. The outline presented in this article incorporates the newly evolving knowledge, skills, and abilities needed to provide expert pediatric nursing care. PMID- 12087652 TI - New initiatives in end-of-life care. PMID- 12087653 TI - Andrea Yates: where did we go wrong? PMID- 12087654 TI - Postpartum depression: practical advice from two nurse practitioners. PMID- 12087655 TI - Postpartum depression: voice from a historian. PMID- 12087657 TI - [Physiologic significance of the interaction between timolol and free amino acids in eye structures]. AB - The authors analyze the interaction of timolol maleate (Timoptol, Leciva Prague) with free amino acids with regard to the effect on intraocular pressure. The different colour reaction of an amino acid mixture with Timoptol, the decline of radioactivity of Na[I125] tyrosine with Timoptol and the formation of 8 radioactive fractions after addition of Timoptol to Na[I125] indicate: a) the specificity of interaction of each amino acid with Timoptol; b) suggest that between Timoptol and free amino acids a firm covalent bond develops with the concurrent development of a new biologically active metabolite. Evidence of the presence of a new metabolite is important from the aspect of understanding and more accurate definition of the action of Timoptol in tissue structures of the eye and thus also for possible effective treatment of glaucoma. PMID- 12087658 TI - [Implantation of the AcrySof MA30BA lens using the Monarch system]. AB - The authors describe experiences with implantation of the intraocular lens AcrySof MA30BA by the Monarch IOL delivery system. They consider the facility of Monarch's system completion, the location of the lens in the cartridge and its folding, lens passage through the cartridge and its expansion intraocculary. The authors evaluate the width of incision necessary for the conduction of injector, preoperative and postoperative course, final visual acuity and inducted astigmatism. The results of the work show that implantation of the lens AcrySof MA30BA by the IOL Monarch delivery system is easy and safe. The width of incision after implantation performs criterion of the no stitch technique and corresponds contemporary requirements of the modern cataract surgery of small incision. PMID- 12087659 TI - [Calculation for emmetropic intraocular lenses in hypermetropia]. AB - Authors evaluated postoperative refractive errors after implantation of intraocular lens at the set of 429 eyes. There predominated eyes with short axial length (AL) at this set: 304 eyes had AL < 22 mm, average AL was 21.54 mm. The basic point was to correct the linearity of SRK formula by different way from SRK II formula, while keeping the new formula simple to use. Authors reached following modification of A-constant in SRK formula: new A-constant = original A constant + C, where C = 0,146*AL2 - 7,702*AL + 100,130. PMID- 12087660 TI - [Best's vitelliform dystrophy with complications of a neovascular membrane and hemorrhage]. AB - The objective of the paper is to draw attention to an uncommon picture of Best s vitelliform dystrophy of the macula (BVD) complicated by a choroidal neovascular membrane (CNVM) and haemorrhage. In an 8-year-old boy where incidentally at the age of 5 years cystic changes in both maculars were found and a reduced central visual acuity (CVA) of the left eye, the CVA of the so far asymptomatic right eye suddenly deteriorated. In the right macula a vitelliform focus was present the lower half of which was formed by a vascularized scar with subretinal haemorrhage on the margin. In the left macula was a round chorioretinal scar with hyperplasia of the pigmented epithelium (RPE) and remnants of macular yellow. Fluoroangiographic examination (FAG) visualized classical CNVM, optical coherence tomography (OCT) ablation of the RPE. The electrooculogram (EOG) was bilaterally reduced, CVA of the right eye was 6/60, of the left eye 6/36. Examination of the mother revealed though the ophthalmological finding was normal, a significantly reduced EOG and in an older brother there was a small vitelliform focus in both maculae and reduced EOG with CVA of 6/6. CONCLUSION: CNVM with haemorrhage is a rare complication of BVD. This complication markedly extends the polymorphism of findings, is a risk of the CVA and makes the diagnosis more difficult. EOG examination is decisive for the diagnosis of less common forms of BVD. PMID- 12087661 TI - [Use of computer tomography for localization of a biopsy needle for tissue collection from the orbit]. AB - In four patients with orbital expansion the authors used computed tomography (CT) to locate the bioptic needle inserted into the orbit. In all these instances it proved possible to collect a sufficient amount of pathological tissue to assess the diagnosis. Serious side-effects were observed only in one case, where the patient developed after insertion of the needle transient bradycardia. CT was also used to locate the needle for exhausting of blood from a lymphangioma of the orbit. PMID- 12087662 TI - [Eye injury from a paintball projectile]. AB - A fourteen-year-old adolescent suffered after a direct hit with a plastic projective (paintball) a severe injury of the anterior and posterior segment of the eye manifested by intraocular haemorrhage, cyclodialysis, detachment of the retina with two giant tears and oedema and haemorrhages of the retina. After cerclage, pars plana vitrectomy and transient four-month tamponade with silicone oil the retina reattached. Late complications, cataract, broad anterior adherence with scars of the chamber angle and ciliary body, a lamellar defect of the macula and partial atrophy of the optic disc determined the subsequent development and were an indication for cataract surgery, implantation of an artificial lens into the lenticular capsule and reconstruction of the pupil. The final result was from the cosmetic and functional aspect (visual acuity 0.3) very satisfactory. Plastic projectiles (paintballs) are a new cause of severe eye injuries. At risk are in particular participants of games who do not protect their eyes with spectacles or masks. To save the function of the eye in unnecessary injuries frequently several operations are needed and close collaboration of surgeons for the anterior and posterior segment. PMID- 12087663 TI - [Aniridia]. AB - Aniridia is a congenital developmental anomaly of the eye that usually affects both eyes. The development of the iris, cornea, lens, angle, optic nerve and retina is disturbed. Aniridia is most often a hereditary disease with an autosomal dominant, rarely autosomal recessive inheritance, but sporadic cases are also possible. The vision function in aniridia has been observed to have a wide range from blindness to a normal visual acuity. The more serious cases where blindness occurs has been due not specifically to the aniridia but to associated conditions like cataract, glaucoma, foveal hypoplasia, corneal dystrophy, nystagmus. Aniridia could be associated with the mental retardation. Some of the sporadic cases develop Wilms' tumor, frequently as part of the WAGR syndrome (Wilms' tumor, aniridia, genitourinary abnormalities and mental retardation). PMID- 12087664 TI - [Antimetabolites in glaucoma surgery--long-term results]. AB - PURPOSE: To observe the long term success of antimetabolites used in glaucoma filtering surgery in eyes with high risc of failure. MATERIAL AND METHOD: Two groups of patients undergone trabeculectomy with the use of antimetabolites 5 Fluorouracil (5-FU) and Mitomycin C (MMC) are included in this retrospective study. 5-FU was administered postoperatively 1-10 times in the dose of 5 mg/ml in subconjunctival injection on 32 eyes. Follow up time ranged from 8 months to 9 years. Mitomycin C was administered intraoperatively in the dose of 0.2 mg/ml, on subconjunctival sponge with application time from 3 to 5 minutes on 15 eyes. Follow up time ranged from 7 month to 4.5 years. RESULTS: Intraocular pressure (IOP) before operation with the use of 5-FU was in average 28.8 mmHg (22-50 mmHg) at the end of therapy with 5-FU was 17.6 mmHg (10-30 mmHg) and in the last examination was on average 19.5 mmHg (14-28 mmHg). IOP before operation with the use of MMC was in average 30.3 mmHg (22-40 mmHg), the seventh day after administration of MMC 12.7 mmHg (6-20 mmHg) and in the last examination was in average 18 mmHg (9-28 mmHg). During the long term observation of IOP values in the 5-FU group there was 79.2% success and in the MMC group there was 83.3% success (IOP up to 20 mmHg with or without therapy). Decrease of IOP was statistically significant. CONCLUSION: Antimetabolites improve the success of filtering surgery in high risc glaucomas. The long term follow up shows that failure of filtration in these types of glaucomas is still a serious clinical problem. PMID- 12087665 TI - [Computer perimetry--rapid TOP (tendency oriented perimetry) and normal threshold methods in clinical practice--comparison of results]. AB - Computer perimetry is constantly being developed but still remains a subjective examination method. The patient's cooperation plays an important part and therefore examination strategies are improved to make it possible to obtain the most accurate possible results during the shortest possible examination time. The author presents an account on the comparability of results of rapid TOP strategy (Tendency Oriented Perimetry) and normal threshold strategy with programme G2 on the perimeter OCTOPUS 101. It involves the retrospective evaluation of 73 visual fields which were divided into four groups according to the achieved retinal sensitivity during examination with TOP strategy and with normal threshold strategy. The author compared the obtained parameters MD and LV, she compared the sites of defects of retinal sensitivity in both strategies and calculated the time saved by rapid TOP strategy. High comparability was achieved in group 1 with normal retinal sensitivity in both strategies and also in group 4 with abnormal retinal sensitivity in both strategies. In groups 2 and 3 comparability was not proved (group 2--abnormal retinal sensitivity in TOP and normal retinal sensitivity in normal threshold strategy, group 3--normal retinal sensitivity in TOP and abnormal retinal sensitivity in normal threshold strategy). In these groups we may assume a higher ratio of the so-called learning effect or conversely fatigue and poorer cooperation of the patients during the lengthy examination by normal threshold strategy. The localization of defects agreed in group 4 in 83.3% and in the other groups it was not compared (group 1 practically without defects, in groups 2 and 3 the results were not comparable). The total time saved with TOP strategy was 75.8%, the mean examination time with TOP strategy was 3 minutes 27 seconds, with normal threshold strategy 14 minutes 17 seconds. With regard to the significant time saving the programme G2 with TOP strategy can be considered suitable for patients with glaucoma, with intraocular hypertension and with suspected glaucoma but it can be also used as a screening method with quantitative results for other situations. PMID- 12087666 TI - [The Kayser-Fleischer ring and its diagnostic importance]. AB - This paper described Kayser-Fleischer ring as an early diagnostic sign of the Wilson's disease. This disease is demonstrated in case of 37-years old white man, a member of affected family. We discuss about typical manifestations of the disease in the eye and especially about Kayser-Fleischer ring with the use of the slit lamp biomicroscopy, spectrophotometry and electron-microscopy. PMID- 12087667 TI - [Screening for threshold stages in retinopathy of prematurity]. AB - The authors evaluated retrospectively the examination protocols of 208 children (400 eyes) treated by cryocoagulation on account of active retinopathy of premature infants (ROP). The objective was to assess the optimal timing for the onset of examination to record in time the threshold stages of ROP in children with different grades of maturity. The baseline point was the finding that before the 5th postnatal (pnt) and 31st postconception week (pcw) the threshold stage was not diagnosed in any child of the group. The highest incidence of threshold stages in all children was between the 36th and 37th postconception week. The first examination made between the 5th-6th postnatal or at first during the 31st postconception week is sufficient to cover initial threshold stages of ROP in children with different birth weights. Screening based on combination of the postnatal and postconception age is more accurate than evaluation by one criterion. Premature examinations are misleading and not valid for screening. PMID- 12087668 TI - [Dacryolithiasis--analysis of 15 cases]. AB - Dacryoliths are found in 15-20% patients subject to dacryocystorhinostomy. Dacryoliths are formed by proteins or calcium and phosphate salts. In the development or dacryolithiasis participates chronic inflammation of the sac, it is found more frequently in women where some part is played by the structure of the lacrimal duct, hormonal influence and the use of cosmetic preparations. A typical sign of the disease in the variability of complaints and results of clinical examinations. Assessment of the diagnosis before surgery can be facilitated by dacryocystography. The method of choice in dacryolithiasis is dacryocystorhinostomy. The authors present their own group of 16 dacryolithis which they observed 10x in women and 5x men. The concrements were examined by the method of infrared spectroscopy and gas chromatography. All were formed by proteins; calcium and phosphate salts were not found In all patients after dacryocystorhinostomy the complaints disappeared. PMID- 12087669 TI - [Polymorphonuclear leucocytes and the production of reactive oxygen species after total hip and knee replacement]. AB - Aseptic loosening of the prosthesis is caused by the mechanical instability of it's components or it depends on direct tissue reaction--foreign body reaction to implant's wear debris. Former studies showed a significant role of macrophages in the process of the chronic inflammation leading to the aseptic loosening. There are few studies concerning the early reaction against the implant and concerning in this process the role of neutrophils--the cells which form the primary defence of the body and are activated during contact with the foreign body. The examined group consisted of 33 patients aged 59-79 with implanted cemented hip or knee endoprosthesis due to the primary osteoarthritis. The count of granulocytes, production of the nitric oxide and hydrogen peroxide with Pick's and Market's method was evaluated in samples collected before the surgery, 2 months and 3 years after the operation. 3 years postoperatively 8 patients reported pain of the operated joint and in two cases aseptic loosening was recognized. In this group the level of production of the nitric oxide and hydrogen peroxide by the neutrophils 2 months postoperatively was significantly higher in comparisons to the levels in the patients without pain and the control group. In patients without pain the production of nitric oxide and the hydrogen peroxide did not change significantly after the surgery. The presented results suggest the neutrophil's activation by the cytokines, which appear as a result of a local response to a foreign body, may be a factor initiating a chronic inflammation around the implant leading to the aseptic loosening. PMID- 12087670 TI - [PSO modular hip endoprosthesis--personal experiences]. AB - Hip prosthesis reimplantation, particularly with concomitant bone destruction around the stem, leading to a total loss of mechanical properties of the joint, is a major concern of modern orthopedics. Between may 1998 and August 2000 nine reimplantation procedures were performed in our Department. All were performed with the use of the Biomet PSO system. In four cases reimplantation was performed because of aseptic loosening of the prosthesis, with massive bone destruction around the stem. In three cases surgery was necessary because of loosening of reimplanted prosthesis. In two cases PSO prosthesis was used because of intertrochanteric fracture instability concomitant with massive femoral head and intertrochanteric bone destruction. Functional assessment was based on the Merle d'Aubigne classification. On 6 points were attained before surgery and 10 points- 6 months post-up, the difference resulting from a decrease of pain. Luxation of the prosthesis was noted in two cases one 15 days after surgery and the other one 6 months post-up. In our opinion use of PSO endoprosthesis is a very valuable method of treating cases with bone destruction of the femur following loosening of hip prosthesis. PMID- 12087671 TI - [Internal fixation in the treatment of radial head fractures]. AB - The authors give indications for internal fixation of the radial head fractures. The results in 20 patients, age 18-69 (average 38 years), 12 male and 8 female have been reported. The authors conclude that fixation is a better method of treatment than resection of the radial head. The authors suggest that good fixation is also possible in the case of comminuted fracture. Fracture dislocation of the elbow joint is not an indication for ligaments repair if the elbow is stable after radial head osteosynthesis. PMID- 12087672 TI - [Monitoring of the early stages of bone healing process based on serum osteocalcin level changes]. AB - The aim of this paper was assessment of utility of serum osteocalcin levels in monitoring early stages of bone healing and early detection of bone healing disorders. Serum osteocalcin level and alkaline phosphatase activity was assessed with acute long bone fractures and with delayed bone union and pseudoarthroses. 25 patients underwent the trial. The analysis of the results confirmed the utility of serum osteocalcin level as a prognostic factor in early detecting of union disorders and its correlation to radiological examination. PMID- 12087674 TI - [Acute carpal tunnel syndrome after distal radius fractures--long term results of surgical treatment with decompression and external fixator application]. AB - Acute Carpal Tunnel Syndrome (ACTS) is a relatively rare sequel of distal fractures of the radius. This paper is an analysis of the causes of ACTS and of long-term results of treatment by surgical decompression combined with use of external fixators. Out of 128 patients with distal radius fractures treated in our department 11 cases required surgical treatment because of ACTS. Fractures were classified according to the AO classification as C2 and C3. CTSA was diagnosed basing on typical clinical signs: pain numbness over the radial three and a half fingers. All patients were treated surgically by reduction of the fracture, external fixation and decompression of the median nerve, all performed simultaneously. The results were assessed within 11 months (on average) postoperatively. It was found that ACTS depends on the severity of the fracture (comminuted, dislocated) and on method of treatment (local anaesthesia, multiple reductions, Cotton-Loder position). Long term results: pain was absent the two point discrimination test was within norm, light sensation was within norm in 9 cases and slightly decreased in 2 cases. There were no positive Phalen tests and no Tinel signs. Grip strength raged from 17 to 120 kg (mean: 38 kg). According to Gartland-werley classification 5 excellent, 3 good, and 3 acceptable results were obtained. This indicates that early recognition and prompt treatment by surgical decompression and external fixation results in significant improvement of symptoms and prevents persistence of symptoms. PMID- 12087673 TI - [Diagnostics and treatment of intervertebral disc herniation]. AB - The paper reviews procedures for treatment of intervertebral disc herniation in our Department, evaluating modern scanning methods and surgical procedures. A total of 173 cases aged 13-73 years are presented. Discectomy was performed at surgery, with recent cases also embracing intervertebral spondylodesis with the use of cages to restore intervertebral distance. Short-term follow-up of patients after surgery have been very encouraging. PMID- 12087675 TI - [Results of surgical treatment of congenital clubfeet with the Sotirow method]. AB - Clinical and radiological results of surgical treatment of congenital clubfeet by an antero-lateral approach first introduced by Sotirow in 1976 are presented. The aim of this paper was to assess long-term results of this procedure on children (aged 5-18 months at the time of surgery). A series of 65 feet in 42 cases diagnosed with congenital talipes equinovarus was evaluated. Forty-two cases were treated conservatively with casts--changed on a weekly basis for 2-3 months and then every 2 weeks. Magone's score criteria were used to assess the method. Average follow-up was 7 years. Clinically 72% of the results were rated as excellent or good. We found the biggest problem during the Sotirow procedure was placing the trochlea of the talus bone into his anatomical into its' anatomical position in the talo-crural joint. The method introduced by Sotirow seems to be highly effective and worth applying. It enables a real elimination of soft-tissue contractures, which retains the deformity of the foot through an antero-lateral approach. PMID- 12087676 TI - [Experimental and clinical of the range of the posterior tibial island flap]. AB - Basing on anatomical studies (20 dissected lower extremities) and clinical experiences (4 cases), the paper presents local transposition possibilities of a distally pedicled tibial flap (pedicled on the posterior tibial artery). The flap allowed for soft-tissue coverage of 2/3 of the plantar side of the foot and 1/4 of the dorsal aspect of the foot. PMID- 12087677 TI - [The treatment of acute hematogenous osteomyelitis in children and young adults]. AB - On the basis of own experiences and literature from last three years authors presented the role of surgical treatment in children above two years and young people, suffering from acute haematogenous osteomyelitis. Conservative treatment should be limited to these cases, where we can see a quick recovery, reducing of the pain and decreasing of the level of CRP and ESR. The presence of subperiosteal abscessus, positive bacteriological culture from blood and other complications (i.e. staphylococcal pneumonia), are the absolute indications to decompression of the medullary cavity, and local application of antibiotics. PMID- 12087678 TI - [Osteomyelitis concomitant with psoriasis]. AB - The paper presents a case of child suffered from osteomyelitis concomitant with psoriasis. Similar clinical and radiological course was observed in another two cases. The disease was chronic, recurrent and school children were affected. Normal temperature and increased blood sedimentation were observed. Enlarged of metaphysis, "marbled" structure of the bone with many little osteolitical focus and thickening of cortical bone were observed, too. This disease was progressive and the whole of the diaphysis was affected. Pus was never found during operation. The bone was fragile sclerotic in some places, with thickened periosteum. Histopathological examination indicated chronic osteomyelitis. Bacteriological examination was always negative. The treatment performed usually in acute osteomyelitis wasn't effective. PMID- 12087679 TI - [Pathogenesis of Dupuytren's contracture--a review]. AB - Dupuytren's disease is a palmar fibromatosis bringing about irreversible finger contracture. Histopathologically, the disease is characterized by the presence of the two types of structures: nodules, containing of intensively proliferating cells, and fibrous cords, formed by thick bundles of collagen fibers. It seems that key role in the development of Dupuytren's contracture play alterations of palmar fibroblasts activity. These cells begin intensively proliferate and transform to myofibroblasts. The later ones sharing phenotypic features of fibroblasts and smooth muscle cells take part in remodelling of extracellular matrix and are a source of palmar contracture. The pivotal factors involved in changes of palmar fibroblasts functions seem to be growth factors (mainly TGF beta, PDGF and bFGF). However, the participation of reactive forms of oxygen in mentioned process is also considered. PMID- 12087680 TI - [Reconstruction of tarsus and metatarsus with free iliac flap--a case report and review of literature]. AB - Procedure and late results of treatment of 17 year old female with severe trauma of right foot by agricultural machine are presented. Defects included soft tissues of the dorsal foot, II-IV metatarsal bones, cuneiform bones and majority of the navicular bone. The patient was first treated in a regional hospital and due to infection an amputation of the lower extremity distal part was considered. In our department an emergency necrectomy was carried out, general and local antibiotics were administered and the wounds were split skin grafted. Complete one stage reconstruction of the soft tissues and bones with free osteocutaneous iliac flap was performed three months later. Deep circumflex iliac vessels were anastomosed end-to-side to the only uninterrupted posterior tibial vessels. Two axial K-wires were used to stabilize vascularized bone graft. A fragment of the ilium 8.5 x 6.0 x 1.5 centimeters of favourable arcuate shape was used to reconstruct longitudinal and transversal arches of the foot. The vascularized bone graft healed three months after the reconstruction. The patient recovered full weight bearing capability after a year. No complications were observed in the donor site in spite of the fact that the patient was pregnant and had a normal uncomplicated delivery. PMID- 12087681 TI - [Possibilities and profits of blood saving in orthopedics and traumatology]. AB - The author presents his own experiences related to blood saving methods in orthopedics and traumatology. The analysis was carried out on 56 patients who underwent major orthopedic procedures: 35 total hip arthroplasties, 19 total knee arthroplasties and two fusions of fractures of the spine performed between the end of 1998 and the beginning of 1999. Patients were prepared for elective surgery by carrying out autotransfusion, which gave on average 2 units of blood for each patients. During the post operative period blood for retransfusion was obtained through active drainage using CBC Consta VAC (Strayker) equipment and through active drainage using standard Baxter transfusion bags. This gave approx. 350-700 ml of blood after hip arthroplasties, 500-800 ml after knee arthroplasty and 500 ml after stabilization of spine fractures. An analysis of the morphotic values of blood from CBC (Strayker) and the transfusion bags (Baxter) did not reveal any statistically significant differences. This confirmed that both methods are equally useful for obtaining blood from the post-operative wound. However, the low cost and simplicity of downward drainage make this method more suitable for our hospitals. Autotransfusion yielded in our department an annual saving of 250 l of blood and in many planned operations eliminate the necessity for allogenic blood transfusion. PMID- 12087682 TI - [Natural history of anterior cruciate and medial collateral ligament healing- literature review]. AB - The review discusses natural history of ligament healing of the knee. Medial collateral ligament heals by progressing through a series of inflammatory, proliferative and remodeling phases. Anterior cruciate ligament, after rupture, appears to retract initially, with no evidence of bridging scar formation. Why anterior cruciate ligament does not heal after rupture still remains unclear. PMID- 12087683 TI - A comparison of objective measures of sexual arousal and interest: visual reaction time and penile plethysmography. AB - This study assessed the reliability and validity of a visual reaction time (VRT) measure of sexual interest (G. G. Abel, J. Huffman, B. Warberg, & C. L. Holland, 1998) and the penile plethysmograph (PPG) with audio stimuli. A sample of 57 sex offenders incarcerated at a high-security military prison completed physiological and self-report measures of sexual interest, including the VRT and the PPG. Results indicated adequate internal consistency for both measures. Convergent validity and an assessment of clinical usefulness for both measures indicated that (a) both measures accurately identified offenders against young boys; (b) the VRT, but not the PPG, significantly identified offenders against adolescent girls; (c) neither measure reached statistical significance in identifying offenders against adult women; and (d) the VRT did not reach statistical significance in identifying offenders against young girls and the PPG did reach statistical significance but in the opposite direction as was expected (i.e., men with female child victims had significantly lower arousal to female child stimuli than did men in other victim choice categories). PMID- 12087684 TI - Characteristics of perpetrators of child sexual abuse who have been sexually victimized as children. AB - This paper reports on a study of all convicted child sexual abusers in S.E. London. Almost half the offenders reported experiences of sexual victimization in childhood, and the authors aimed to explore the differences between the abused and nonabused child sexual abusers. A range of background and offending variables were examined. It was found that sexually victimized child abusers were significantly more likely to have experienced a range of childhood abuse and associated difficulties; they were more likely to have a range of psychosexual difficulties, to report greater levels of deviant offence-related attitudes on psychometric measure, to be recidivists, and to offend against boys. Both emotional abuse/physical neglect in childhood and having homosexual contacts in adulthood significantly contributed to a predictive model, reliably distinguishing between sexually victimized and nonsexually victimized child sexual abusers. The implication of these findings for treatment approaches for sex offenders is discussed. PMID- 12087685 TI - An evaluation of the Empat as a measure of victim empathy with civilly committed sexual offenders. AB - Enhancement of victim empathy is a common component in the treatment of sexual offenders. However, consensus about the definition of empathy and the best instrument to measure empathy in sexual offenders is lacking. This paper evaluates the Empat as a measure of empathy with patients civilly committed under California's Sexually Violent Predator law. Rapists and child molesters in the current sample scored significantly higher in victim empathy than any of the original sex offender groups, with rapists scoring higher than the control group cited by M. McGrath, S. Cann, and R. Konopasky (1998). Clinicians have found the Empat to be too face valid and easily manipulated by patients within this population. The construct of empathy as applied to sex offenders is reviewed and the properties of a more adequate measure of this construct are suggested. PMID- 12087686 TI - A comparison of modified versions of the Static-99 and the Sex Offender Risk Appraisal Guide. AB - The predictive validity of 2 risk assessment instruments for sex offenders, modified versions of the Static-99 and the Sex Offender Risk Appraisal Guide, was examined and compared in a sample of 258 adult male sex offenders. In addition, the independent contributions to the prediction of recidivism made by each instrument and by various phallometric indices were explored. Both instruments demonstrated moderate levels of predictive accuracy for sexual and violent (including sexual) recidivism. They were not significantly different in terms of their predictive accuracy for sexual or violent recidivism, nor did they contribute independently to the prediction of sexual or violent recidivism. Of the phallometric indices examined, only the pedophile index added significantly to the prediction of sexual recidivism, but not violent recidivism, above the Static-99 alone. PMID- 12087687 TI - The child sexual abuser: perceptions of college students and professionals. AB - College students and members of the Association for the Treatment of Sexual Abusers (ATSA) were compared as to their beliefs and attitudes concerning perpetrators of child sexual abuse. Analyses of a 44-item inventory (assessing beliefs about an abuser's demographics and attitudes concerning an abuser's cognitions and behaviors) indicated that the groups differed on perceived demographic descriptors (e.g., students believed perpetrators to be older when they first begin offending, more educated, and more likely to be gay than the professionals) and behaviors (e.g., students believed that the perpetrator was more likely to use force to gain the child's compliance). In addition, 2 subscales (Cognitive Distortions and Perceived Social Functioning) were identified. Compared to professionals, students were less likely to believe perpetrators use cognitive distortions and were more likely to believe perpetrators function at a lower interpersonal level. Results are discussed in terms of the efforts to educate the public about the characteristics of child sexual abusers. PMID- 12087688 TI - Leadership in turbulent times is spiritual. AB - In the past decade, two formerly estranged domains, spirituality and work, have been increasingly linked. Why has this happened? This articles posits that this strange union is an unavoidable consequence of living in a turbulent time. As our world grows more chaotic and unpredictable, leaders must respond to questions that have historically only been answered through spiritual traditions: How do I cope with incertainty? How do I help others find meaning in their lives? What are my values? How can I act with courage and integrity? Leaders also play a key role in helping people recognize that life is cyclical, unpredictable, and unstable and that they must engage it as such. This is well-explored in spiritual traditions, but not in traditional management literature. Several organizational and personal practices are offered as simple ways to develop leadership capacity for leading in turbulence. PMID- 12087689 TI - The intersection of analysis and gestalt. PMID- 12087690 TI - Spirituality and effective leadership in healthcare: is there a connection? AB - Does the level of a leader's spirituality affect his or her effectiveness as a leader? The existence or nonexistence of a link between leadership and spirituality is an emerging issue in the healthcare leadership and management literature. This article defines effective leadership and spirituality and reviews the literature for both healthcare and other organizations in the discussion of this link. It concludes that healthcare leaders who are more developed in terms of their actualized spirituality simultaneously implement the five practices of effective leaders: challenge the process, inspire a shared vision, enable others to act, model the way, and encourage the heart. Moreover, they achieve more positive results for their organizations. PMID- 12087691 TI - Considering spirituality: its promise and purpose. PMID- 12087692 TI - Leadership and spirituality: a response. PMID- 12087693 TI - Preparing your PFS Department for HIPAA. PMID- 12087694 TI - [Gilbert Schogel, "From the other side of the screen"]. PMID- 12087696 TI - [Winckler's films. Interview by Helene Delmotte]. PMID- 12087697 TI - [Our nurses, our heroes]. PMID- 12087698 TI - Miami Children's Hospital creates new Web database. PMID- 12087699 TI - Children's hospitals establish patient pages for parents. PMID- 12087700 TI - Should doctors prescribe information along with pills? PMID- 12087701 TI - Web strategies of health plans and health insurance companies. PMID- 12087702 TI - Mercy Hospital aims Webcast of surgery at consumers. PMID- 12087703 TI - Schizophrenia. AB - Schizophrenia is a biologically based mental illness that may manifest in hallucinations. Drug treatment and social interventions may control symptoms. PMID- 12087705 TI - Conservative sharp debridement: the professional and legal issues. AB - In recent years there has been much debate over the use of conservative sharp debridement in the treatment of certain wound types. Here it is discussed in relation to increasing the speed of wound healing in slow-to-heal wounds. The authors examine education and skill competence in relation to the professional's duty of care to patients with wounds. PMID- 12087704 TI - Establishing nutritional guidelines for critically ill patients: Part 1. AB - Patients who are critically ill can become malnourished very quickly, which has an impact on their recovery and length of hospital stay. In the first part of this paper on producing nutritional guidelines, the author presents her rationale for the guidelines and examines the effects of malnutrition, risk assessment, interventions available and the ethical considerations. PMID- 12087706 TI - Beta-adrenoceptor blocking agents. PMID- 12087707 TI - Care of patients with brain injury in the critical care environment. AB - Nurses caring for patients with brain injuries need to be aware of the effect of such injuries on brain anatomy and the problems associated with a rise in intracranial pressure. This paper examines the main aspects of nursing care for brain-injured patients. Further research is required in many areas of management in this patient group to provide up-to-date, evidence-based care. PMID- 12087708 TI - Improving a nurse-led flexible cystoscopy service through audit. AB - The urology nurse practitioner in one trust expanded her skills to perform flexible cystoscopy. This has reduced waiting times for patients and has led to improved service delivery and continuity of care. Auditing of the protocol used ensures there is a high, consistent standard of follow-up care for all patients. PMID- 12087709 TI - The implications of improvements in screening for prostate cancer. AB - With Men's Health Week running from 10 to 16 June this year, there is an increasing focus on men's health. Prostate cancer may be traditionally associated with elderly men, but with improvements in screening techniques increasing numbers of patients are being diagnosed at a younger age, perhaps changing the view of this disease. PMID- 12087710 TI - Breaking bad news in cancer care. AB - When a diagnosis of cancer, such as prostate cancer, is made nurses may be involved in presenting the news to the patient. How that news is given can make the experience a distressing or reassuring process. In the first part of this paper, the authors examines the human qualities required by professionals and the barriers to breaking bad news. PMID- 12087711 TI - Tackling the problem of the social exclusion of teenage mothers. AB - The majority of teenage mothers in the UK come from poor backgrounds, with few prospects of work locally and little to aim for educationally. But with help from government education and child-care schemes and the intervention of health-care professionals, much can be done to improve their position. This paper looks at current research and practical projects in this area. PMID- 12087712 TI - Evaluation of community nurses' knowledge of diet for diabetes. AB - A questionnaire-based study to ascertain community nurses' knowledge of dietary recommendations for people with diabetes mellitus identified several gaps, a problem acknowledged by many of the participants. This information was used to produce a pocket guide to advising people with diabetes on their diet for use by community nurses. PMID- 12087713 TI - Providing written information for patients with head and neck cancer. AB - Nurses in one Northern Ireland hospital asked patients, their relatives and members of the multidisciplinary care team for their input in compiling an information booklet for patients undergoing major surgery for head and neck cancer, with the aim of creating an information source which would be attractive to a range of patients with head and neck cancer. PMID- 12087714 TI - Staff attitude surveys. The morale majority. AB - Since April 2000, it has been mandatory for all NHS organisations to conduct annual surveys of staff morale. Analysis of staff attitude surveys in 18 trusts and 20 health authorities in 2001 showed that most staff groups were not happy with their pay or the recognition they received. Managers were happy with all aspects of their jobs. Doctors and nurses were concerned about their safety. All employing organisations must act on these results to improve employment practice and help to develop motivation. PMID- 12087715 TI - Open space. Power with responsibility. PMID- 12087716 TI - Primary care trusts. Leading lights. AB - Involving clinicians, lay members and managers on the boards of primary care trusts has great potential for producing innovative services. The involvement of clinicians means decisions are more likely to be acceptable to staff on the frontline. It is important that strategic health authorities foster innovation and do not seek to become controlling of PCTs. It is vital that a culture of performance management does not hamper PCTs' development. PMID- 12087717 TI - [Effect of peptide antibacterial factor and changes in Staphylococcus susceptibility to antibiotics]. AB - The susceptibility to antibiotics of the Staphylococcus epidermidis cells in the presence of low-molecular weight peptide factor was investigated. The factor was isolated from the Styaphylococcus warneri culture media. The factor enhanced the S. epidermidis susceptibility to ampicillin, chloramphenicol, lincomycin, rifampicin, tetracycline, cefalexine, erythromycin and fusidine. The same effect was demonstrated for the cells of S. epidermidis resistant to cadmium ions along with resistance to several antibiotics. However the cells reaction in this case was expressed to a lower extent. It was suggested that revealed alterations in staphylococci susceptibility to antibiotics was due to the peptide factor effect on cytoplasmic membrane permeability that resulted in nonspecific increase in accumulation of various compounds with low-molecular weight. PMID- 12087718 TI - [Eradication of Helicobacter pylori in patients with duodenal ulcer following the short course of treatment with azitromycin and amoxycillin]. AB - The efficacy of short-term treatment with azithromycin in 17 patients with acute doudenal ulcer associated with H. pylori was evaluated. Bioptats of the gastric mucosa taken at the beginning and after one month of treatment were investigated for H. pylori presence by histological, bacteriological methods, by urease test and by PCR. All the patients with positive H. pylori test were treated with Gastrozol (omeperazole, ICN Pharmaceutical) 40 mg per day for 1 week followed by 20 mg per day for 3 weeks, Sumamed (azithromycin, peira) 0.5 g once daily for 3 days and amoxycillin 0.5 mg four times a day for 10 days. Bioptats analysis before treatment revealed H. pylori in 100% for PCR methods, in 94.1% for urease test, in 88.2% for histological test. After the treatment H. pylori was revealed in 12.5% for urease and histological test, in 18.8% for bacteriological test and in 25% for PCR test. Thus the treatment efficacy was 75%. Side effects for short term azithromycin therapy were shown in 5.9% cases. PMID- 12087719 TI - [Glutoxim in the complex treatment of tuberculosis]. AB - Results of Glutoxim investigation are presented. Glutoxim is the the drug of the new class--thiopoietins. It is considered to be immunorehabilitator as it modulates intracellular process of thiols metabolism, initiates cytokins system, activates phagocytosis etc. Results of the glutoxim administration at the 42 patients with tuberculosis using traditional treatment regimes are presented. Results of the randomized study at the patients with severe disseminated drug sensitive and drug-resistant pulmonary tuberculosis demonstrated high efficacy of the glutoxim and its good tolerability. Glutoxim administration allowed to shorten the period of tuberculosis intoxication signs disappearance, to shorten the period of sputum negativation and shortened the period of pulmonary inflammation reverse process. Glutoxim application was specially favorable at the patients with severe tuberculosis complicated by viral or medicamental hepatitis. PMID- 12087720 TI - [Perioperative management of focal liver diseases by ofloxacin]. AB - Perioperative use of ofloxacin for prophylaxis was investigated in 20 patients with focal hepatic formations (hemangioma, adenocarcinoma, echinococcosis). First dose of ofloxacin (200 mg) was given intravenously 15 min before operation. After operation ofloxacin was used intravenously (400 mg daily) for 5 days. Pharmacokinetic investigation demonstrated that perioperative intravenous use of ofloxacin provided concentrations in blood and hepatic tissue satisfactory for potential microflore inhibition. Immunological monitoring demonstrated positive dynamics on 5-7 days after operation. dynamics depended on nosology of the focal hepatic formation. Ofloxacin use for prophylaxis in the operated patients with focal hepatic formations was efficient for profilaxy of postoperation infective complications. PMID- 12087721 TI - [Novel beta-lactam structures: design problems]. AB - The methology of the development of new biologically active betalactams is proposed. One of the two ways proposed is specific modification which is peculiar to certain betalactam structure and involves introduction of substitutes changing particular physico-chemical properties of the natural or synthetic analogous. General guidelines for modification of the biologically active compounds are given. The space and depth of the necessary physico-chemical investigation are determined. The alternative way of the new biologically active compounds development is the principle of similarity. The distinctive feature of this approach is the use as building blocks of the substitutes already used in the well-known betalactam antibiotics and "implantation" of this substitutes into other (new) betalactam structures. The ways of the new betalactams synthesis including the methods of enzyme engineering are considered. The possibility to use enzyme engineering processes for production of not only new individual betalactams--hits, but also for synthesis of the groups of betalactams--leads, is shown. More than 6000 new betalactam structures were constructed on the base of the principle of similarity. At least 700 of this compounds demonstrates not only antimicrobial activity but other types of biological activity due to the implementation of additional pharmaceutical units other than betalactams. The constructed compounds are summarized in the tables, the request for the electronic version of the tables can be sent by the address: valan@orc.ru. PMID- 12087722 TI - [Immunomodulating effect of lysozyme in acute cold stress]. AB - It was shown that lysozyme (1 or 5 mg/kg) injection in Wistar rats cooled immersially caused potent immune modulating, antioxidant and hepatoprotector effects. Lysozyme effect was due to heavy erythrocytes and by cytokines induced in spleen (with glass-adhesion at 32-37 degrees C). PMID- 12087723 TI - [Linezolid (Zyvox)--a novel antibacterial agent for treatments of multidrug resistant infections]. PMID- 12087724 TI - [Minimum inhibitory concentrations of various antifungal agents against Basidiomycetes clinical isolates]. AB - The basidiomycete yeasts are often isolated from clinical samples. A minimal inhibiting concentrations (MIC) of ten antifungals of different groups--azols, allilamines, polyens etc.--against isolates of Rhodotorula, Cryptococcus [symbol: see text] Trichosporon yeasts genera were estimated. No one of these cultures was sensitive to azoles at concentrations 0-256 mcg/ml. A vitality of cultures after incubation during 10 days with antifungals was investigated. Miramistin was the most potent fungicidal agent against all cultures. PMID- 12087725 TI - [Premorbid stage in the development of anxiety in children]. AB - The peculiarities of premorbid stage in children, aged 5-15 years, with ICD-10 diagnosis of anxiety-phobic disorders have been studied. The patient's mental state has been described earlier (Savostyanova, 2001). Retrospective evaluation of the patient's state in premorbid allowed to detect the disturbances in personality formation, anxiety phobic reactions and autochthon phases, represented by continuum of subclinical disorders, from floating anxiety to the signs of vital anxiety. These traits are distinguished by instability and inconsistency as well as by mild expression, which did not reach a level of manifest disorders. Personality of the children, who develop anxiety phobic disorders, may be emotionally labile as well as rigid one inclining to negative effectiveness. Hereditable and environment factors influencing anxiety phobic disorders development were detected. The premorbid traits are concluded to reflect inherited anxiety diathesis, which determines the peculiarities of reaction to negative environment and results not only in personality development but also in the large spectrum of future anxiety phobic symptoms. PMID- 12087726 TI - [Dynamics of neurotic depression in children and adolescents]. AB - Seven hundred thirty nine children and adolescents with neurotic depression (293 male, 446 female, aged 1-17 years) have been examined. Depression was mildly expressed in 90.9% of the cases and moderately--in 9.1%. In clinical picture anxious, asthenic, astheno-anxious and anxious-melancholic depression types were present. In neurotic depression in children and adolescents, the initial (the first) and subsequent (the second) stages have been detected. In a case of favorable development of the disease it stops at the initial stage. In a case of poor prognosis (192 patients) the dynamics of disease was distinguished by 3 features: increase of somatization and development of psychosomatic disorders (119 patients), endogenization (45) and formation of pathologic personality changes (28). During treatment clinical variants of disease and its dynamics peculiarities in the every stage are suggested to be taken into account. PMID- 12087727 TI - [Deviation in psychosexual development in the pre-puberty children]. AB - Psychosexual health of 308 children, aged 2-11 years, as well as that of their families, was studied. Deviations in psychosexual development were found in 34.6% of the children examined. The following types were detected: difficulties in formation of gender-determined behavior features--64.4%, precocious psychosexual development--13.7%, delayed psychosexual development--12.3%, obsessive masturbation--9.6%. Risk factors for deviant psychosexual development were found. PMID- 12087728 TI - [Botox in children with cerebral palsy and triceps syndrome]. AB - Botulinum toxin (botox) therapy was used in the children with cerebral palsy. Nine children, aged 3-6 years, with "triceps-syndrome" domination in a clinical picture, have been examined and treated. Control group included 9 children who have not been treated with botox. A three-week rehabilitation course has been prescribed to all the patients. Children treated by botulinum toxin injections performed better back of the foot bending, some electroneuromyographical indices being also optimized. The results obtained imply a use of botox for treatment of children with spastic forms of cerebral palsy. PMID- 12087729 TI - [Transcranial micro-polarization in the treatment of ADHD in children and adolescents]. AB - An effect of transcranial micropolarization on attention deficit hyperactivity disorder treatment in 57 children and adolescences has been studied. A higher efficacy of the method was shown: the patients revealed a significant decrease of impulsivity, hyperactivity and increase of attention (in 71-77% children). PMID- 12087730 TI - [The use of instenon in children with minimal brain dysfunction]. AB - Minimal brain dysfunction (MBD) represents the most common type of neuropsychic disorders in childhood. Resulting in focal damages, underdevelopment and dysfunction of one or another cortical regions of brain hemispheres, MBD manifested in children as movement and speech disorders, dysgraphy, dyslexia, dyscalculia and attention deficit hyperactivity disorder. In the open controlled study, an efficacy of the complex nootropic medication "Instenon" was evaluated in the treatment of 59 MBD patients, aged 4-12 years. Control group included 27 children with MBD assigned to low polyvitamin (Sana Sol) doses. The treatment duration was 1 month. Before and after treatment, children with MBD underwent complex examination, which included parent's interviewing using structured questionnaire, general examination, with detailed analysis of complaints, neurological status investigation and psychological study. In the children taken instenon, distinct positive effect has been achieved in 71% of the cases, in control group--in 15%. Positive effect emerged in improvement of behavior characteristics, better school marks, movement, attention and memory indices, functions of psychic activity, organization, programming and control. When strictly keeping a scheme prescribed (gradual dose increase, drug taking in morning and day time), a risk for unfavorable side effects is minimal. PMID- 12087731 TI - [Post-stroke depression: an experience in using cipramil]. AB - Fifteen patients, 12 males, 3 females, aged 59.6 +/- 9.6 years, with depression after stroke, occurred 9 months--1 year and 3 months before, were treated with cipramil in dose 20 mg/day during 2.5 months. Control group comprised 10 patients matched for demographic and clinical characteristics and been not assigned to cipramil. All patients were fully examined by clinical methods, including computer tomography. Besides, for patient's state evaluation, Hamilton rating scale for depression and Lindmark scale for focal neurologic deficit measuring have been administered. The initial scores on these scales were 22.8 +/- 4.5 and 381.4 +/- 33.4 in the basic group and--22.2 +/- 6.3 and 390.6 +/- 29.8 in the controls, respectively. While cipramil using, an improvement was observed rather quickly--after 2-3 weeks, being stable during the whole treatment course. After treatment finishing, depression scores on Hamilton scale decreased to 15.8 +/- 5.2 (p < 0.001) in the basic group and did not change in the control one. The neurological deficit expression has not been changed. Side effects were mildly expressed in 20% of the patients. In one-third of the patients, antidepressant effect diminished in 1.5 months after treatment finishing. A need for longer antidepressant treatment of post stroke patients is discussed. PMID- 12087732 TI - [Childhood epileptology as an instrument for studying the developing brain]. PMID- 12087733 TI - [Electroencephalographic characteristics of West syndrome]. AB - The study aimed at interictal electroencephalographic characteristics investigation in the patients with West syndrome. It has been carried out in 48 children, aged 3 months--2 years, the inpatients of Psychoneurology and Epilepsy Department, Russian Clinical Children Hospital No. 2, from March 1999 to march 2001. The following EEG awakeness types were detected: typical hypsarrithmia--8 (16.7%) patients; different variants of modified hypsarrithmia--35 (72.9%); a presence of focal epileptiform discharges, but not in the form of modified hypsarrithmia with focal component--3 (6.2%); an absence of epileptiform discharges--2 (4.2%). Typical and modified hypsarrithmia cases ratio was estimated as 18.6%:81.4%. Among the patients with modified hypsarrithmia, the following variants have been detected: synchronized variant of modified hypsarrithmia--35.3% of the cases with modified hypsarrithmia; asymmetric regional or unilateral hypsarrithmia--42.9%; hypsarrithmia with partial component -45.7%; hypsarrithmia with persisting "suppressive-burst" pattern--20% of the cases. Correct evaluation of clinical, electrophysiological and neurovisual data (clinico-electro-anatomical approach) facilitated the rational differential choice of antiepileptic therapy. The approach allows the early prognosis of disease course and its transformation to other epilepsy types. PMID- 12087734 TI - [Membrane destabilizing processes in perinatal nervous system disorders in children]. AB - A complex prospective clinical and biochemical study was carried out in 252 children with perinatal encephalopathy during the 1st year of their life. The lipid peroxidation process, antioxidant system, lysosomal enzymes, phospholipid compound of thrombocyte membranes were studied. The lipid peroxidation process and lysosomal enzymes activation as well as a decrease of antioxidant defense level have been found. Membrane destabilization processes in the children with perinatal encephalopathy correlated with a character of hypoxic influence (acute, chronic, combined), a degree of severity and clinical manifestations of the pathology, and disease stage. Clinical and biochemical efficacy of antioxidant mexidol (0.1-0.2 ml/kg of the body weight intravenously for 10 days) and inhibitor of lysosomal enzymes contrical (1000 U/kg intravenously for 3-5 days) in the infants with perinatal CNS damages is claimed. PMID- 12087735 TI - [Follow-up study of antibodies to nerve growth factor level in children with brain dysgenesis]. AB - A level of antibodies to nerve growth factor (NGF) in blood serum of children with brain dysgenesis has been studied. This level was investigated in follow-up study that allowed to compare it with psychomotor development of the children as well as anatomical brain defect expression (according to MRT and EEG data). Comparing to controls, an increase of antibodies levels in blood serum was found in the patients. No relation of antibodies levels to anatomical brain defect severity was observed. The highest level was shown for the children with the absence of positive dynamics for psychomotor development. In the follow-up study of the children with satisfactory compensation of neurological syndromes, this level decreased to control one. Appearance and elevation of the autoimmune response to NGF can be one of the biological factors, arguing for deterioration of compensatory nervous system potential. PMID- 12087736 TI - [Mental disorders in children abused at an early age]. PMID- 12087737 TI - [Neuromultivit in the treatment of ADHD in children]. PMID- 12087739 TI - [Vascular disturbances in craniovertebral pathology in children and adolescents]. AB - The peculiarities of craniovertebral abnormalities and dysplasias in children and adolescents have been studied. Different diagnostic methods (dopplerography of spinal arteries, ultrasonography, remote infrared thermography, laser doppler flowmetry, etc) were compared. The outlines for treatment the patients with cervical dysplasias are described. PMID- 12087738 TI - [Mental disorders in tumors of maxillary facial region and larynx]. PMID- 12087740 TI - [Neuroimmunopathology: immunoaggression, dysregulation, adoptive immunotherapy prospects]. PMID- 12087742 TI - [Ensemble of politics, knowledge and practice]. PMID- 12087741 TI - [Please do not disturb--I'm nursing!]. PMID- 12087743 TI - [New quality of life as "trouser pilots"]. PMID- 12087744 TI - [Living outside the clinic]. PMID- 12087745 TI - [Nursing quality must improve on a global scale]. PMID- 12087746 TI - [Precautionary "warning blasts"]. PMID- 12087747 TI - [The good old age: learning by doing]. PMID- 12087748 TI - [What do nurses know about health care reforms?]. PMID- 12087750 TI - [A mixture of the modern and the old]. PMID- 12087749 TI - [When patient education allows a new departure]. PMID- 12087752 TI - [The nursing fairy]. PMID- 12087751 TI - [How will our professional identity evolve?]. PMID- 12087753 TI - [Children need closeness]. PMID- 12087754 TI - [In-formation]. PMID- 12087755 TI - [A good process for determining recovery]. PMID- 12087756 TI - [Liberated heads communicate better]. PMID- 12087757 TI - [Immunodeficiency syndrome--with special reference to the recent topics]. PMID- 12087759 TI - [Asthma insights & reality in Japan (AIRJ)]. AB - The asthma insights & reality in Japan survey was aimed to assess the current status of asthma treatment and management in Japan. We used random digit dialing to identify current asthmatics in households throughout Japan. Throughout Japan, 38,132 households were contacted. A detailed survey on asthma management was conducted among 803 of these households, 401 adults and 402 children. More than half had experienced a daytime asthma attack during the previous month. More than one third had experienced an asthma attack at night during that same period. About 70% of the adults and 60% of the children with asthma reported some limitation on activities of daily life. About 50% of adults and 80% of children had never undergone any pulmonary function testing. Only 6% of adults and 7% of children's parents were aware that "airway inflammation" was a major cause of asthma. Moreover, only 12% of adults and 5% of children were using inhaled steroids. There was an especially large gap between subjective perception of asthma control and objective findings in patients with severe asthma. Many Japanese asthmatics tended to underestimate the severity of their condition. Asthma management in Japan falls far short of goals stated in the guideline. These findings emphasize the need for increased awareness of the burden of asthma in Japan. PMID- 12087758 TI - [The effectiveness of the long-term treatment with pranlukast in pediatric patients with mild to moderate asthma]. AB - To investigate the efficacy of the long-term treatment with pranlukast, a specific cysteinyl leukotriene receptor antagonist, in pediatric patients with mild to moderate asthma, 77 pediatric asthmatic patients who received pranlukast for up to 36 months (mean duration, 13 months) were evaluated retrospectively. Treatment with pranlukast resulted in improvements from the pretreatment baseline in asthma attacks per month, episodes of hospitalization, and episodes of intravenous amynophiline treatment on emergent clinic visits. The percentage of responders who had marked or moderate improvements in the above-mentioned parameters of asthma control was 79%. In conclusion, pranlukast caused significant improvements in long-term asthma control in pediatric patients with mild to moderate asthma. PMID- 12087761 TI - [Study of cockroach allergen in adult asthmatics in Japan]. AB - To study the clinical significance of cockroach allergen in adult asthmatics in Japan, we performed intradermal skin test to four commercial base cockroach extracts. The specific IgE antibody to cockroach was measured in sera by CAP RAST. Positive rates of immediate intradermal skin response to cockroach extracts were 10.2-12.6%. Positive rate of CAP-RAST to cockroach was 12.6%. We performed bronchial provocation test on 6 cockroach sensitive asthmatics. Immediate asthmatic response was provoked in 2 cases and late asthmatic response was provoked in one case. Positive histamine release from peripheral leukocytes (HRT) to cockroach was shown in cockroach sensitive asthmatics. RAST-inhibition studies of cockroach showed several inhibition patterns and indicated partial cross reactivity between cockroach species and mite. These results indicate that there is a possibility that cockroach is one of the important inhalent allergens in Japan. PMID- 12087760 TI - [Evaluation before and after pranlukast administration with the QOL questionnaire (revised version 2001) for pediatric patients with bronchial asthma and their parents or caregivers]. AB - We conducted a longitudinal investigation with the QOL questionnaire (revised version 2001) before and after the 4-week-administration of a leukotriene receptor antagonist pranlukast. A significant improvement in the < 4 yrs group was observed at week 1, and that in > or = 4 yrs group at week 2. Under these conditions, the overall QOL score, physical domains and mental domains, significantly improved in both the < 4 yrs group and the > or = 4 yrs group. Overall, a slight correlation was observed between ratio changes in QOL scores and differences in symptom scores. However, no correlation was found in part of patients, suggesting that the QOL questionnaire allows measurement of mental changes in the patients themselves and their parents or caregivers for therapeutic effects which cannot be determined with ordinary physical findings only. In "event present" group, a significant difference in physical and mental domains was revealed by the comparison of QOL scores before and after administration. And furthermore in "event absence" group, the p-value for physical domain and mental domain was 0.0505 and 0.0912 in the < 4 yrs group, respectively, 0.0101 and 0.0446 in the > or = 4 yrs group, respectively. The above results led us to consider the QOL questionnaire (revised version 2001) useful for routine medical care. Furthermore, pranlukast was considered useful for improvement not only of physical symptoms of bronchial asthma but also of the patient's QOL, although the placebo effects in this open trial must be considered. PMID- 12087762 TI - [A case of allergic bronchopulmonary mycosis caused by Schizophyllum commune]. PMID- 12087763 TI - [A fatal case of allergic granulomatous angitis due to rapidly progressing severe intestinal involvement and associated multiple organ failure]. PMID- 12087764 TI - [Recent topics concerning the settlement of the geriatric nursing law (Altenpflegegesetz-AltPflG) in the Federal Republic of Germany]. PMID- 12087766 TI - [Development of a dispositional version of the General Coping Questionnaire (GCQ) and examination of its reliability and validity]. AB - OBJECTIVE: The purpose of the present study was to develop a dispositional version of a stress coping scale (the General Coping Questionnaire, GCQ), especially for university students, and examine its reliability and validity. METHODS: In Studies I to IV, 124, 784, 83, 83 university students respectively completed the dispositional version of the GCQ. The questionnaire has four subscales covering emotion expression, emotional support seeking, cognitive reinterpretation, and problem solving, with a five-point Likert format. In Study IV, in addition to the dispositional version of the GCQ, peer rating was also performed to test the validity. RESULTS: In Study I, 32 items were selected as a second version of the scale, reducing from the first version with 54 items. From Studies II to IV, evidence was obtained for the factorial validity, the internal consistency, the test-retest reliability, the construct validity (except cognitive reinterpretation), and the normal distribution of the scores. CONCLUSIONS: Standardization of the dispositional version of the GCQ with four subscales was essentially completed, facilitating future studies on coping with stress. PMID- 12087765 TI - [The effects of nutrition and life-style on calcaneal bone mass in high school students]. AB - OBJECTIVE: To clarify the effects of nutrient/food intake and lifestyle, especially exercise, on calcaneal bone mass in both male and female high school students. METHODS: A total of 798 healthy Japanese high school students (281 boys and 517 girls) were recruited, and their calcaneal bone was assessed by ultrasound using Lunar Achilles. The subjects were thereby divided into a lower bone mass group and a normal bone mass group according to stiffness at the cut off level of 90 for the boys, and 77 for the girls. Information on lifestyle including frequency of food consumption and regular exercise during the period at junior high school and high school was collected by questionnaire. Nutrient intake was assessed by a semi-quantitative food frequency questionnaire. RESULTS: 1. The intake of total energy and fat in the lower bone mass group was lower than in the normal bone mass group in both sexes. 2. The frequency of subjects not consuming milk in the lower bone mass group was higher than in the normal bone mass group in both sexes. In the boys, the frequency of the subjects who liked to eat green vegetables in the lower bone mass group was lower in the normal bone mass group. 3. The mean stiffness for subjects undertaking exercise during junior high school was higher than in subjects not exercising, even after adjustment for age in both sexes. The frequency of subjects with a habit of sun bathing in the lower bone mass group was significantly lower than in the normal bone mass group in the girls. 4. An analysis using a logistic model was performed to select the factors related to lower bone mass without being confounded with other factors. Lower energy intake in boys and lower fat intake, no exercise during high school life and no milk consumption in girls were each independently related to lower bone mass. CONCLUSION: These results suggest that nutrition and exercise habit affect calcaneal bone mass in high school students of both sexes, especially milk consumption being an important factor related to bone mass in girls. PMID- 12087767 TI - [Factors associated with self-rated health for non-institutionalized aged persons]. AB - OBJECTIVE: To determine factors affecting self-rated health among the non institutionalized elderly in Japan. METHODS: In 20 municipalities in Japan, 6,094 persons aged 65 years and older who were not institutionalized were selected at random. A questionnaire survey was conducted from September through November 2000. The distribution of self-rated health was rated in terms of dependent variables, with odds ratios and their 95% confidence intervals calculated using unconditional logistic models. RESULTS: Of the study population, 5,565 persons (91.8%) responded to the survey. Of the respondents, 64.4% answered that they were healthy, while 28.8% were not. The necessity for periodical visit to hospitals and clinics, and lowering of the activity of daily living greatly lowered self-rated health. Compared with those who were 85 years of age or older, a high health status was observed among those aged between 75 and 84 years, but not those aged 65 to 74 years. Those who tried to have exercise periodically, who had a vice-leadership role in a group, who joined social activities, who lived what they considered a worthwhile life, and who had positive positive activities in daily life, had high levels of self-rated health. Joining social activities for passive reasons elevated the health as well as joining for active reasons such as "because it is fun." CONCLUSION: Even if the reason is passive, joining social activities may elevate the self-rated health levels of elderly people. PMID- 12087768 TI - [Disorders requiring nursing care and the period before recognized as needing nursing care (active life expectancy) in relation to nursing care insurance]. AB - PURPOSE: To measure the state of health of the elderly population, active and dependent life expectancies were calculated based on the number of people needing nursing care. For this purpose, active life expectancy was defined as the period before nursing care was recognized by insurers as being required. Moreover, to cast light on disorders requiring nursing care, age-adjusted nursing time needed for different ailments per ten thousand elderly population was calculated. SUBJECTS AND METHODS: Subjects were those 65 years or over living in Taihaku-ku, Sendai City, recognized as needing nursing care by nursing care insurers. The period before being recognized as needing nursing care was calculated using the Sullivan method, and termed the active life expectancy. Dependent life expectancy = life expectancy - active life expectancy. The number of those needing nursing care caused by each disorder diagnosed by attending physicians, was also age adjusted by the reference population and multiplied by the nursing time needed for each level of nursing, resulting in the age-adjusted nursing time needed per ten thousand elderly population. RESULTS: Those recognized as needing nursing care were 7.5% (7.7% after age adjustment) of the male elderly population, and 12.5% of the female population (10.7% after age adjustment). For men, the active life expectancy was 16.1 years for the age of 65, 9.2 years for 75 and 4.4 years for 85, while the dependent one was 2.0-2.1 years for all ages. For women, the active life expectancy was 19.3 years for the age of 65, 11.1 years for 75 and 4.8 years for 85, while the dependent one was 4.6-5.3 years. The age-adjusted nursing time needed per ten thousand elderly population was 874 hours for men and 1,125 hours for women: of the time 51% was for men with cerebrovascular disease (40% for cerebral infarction), 11% for men with dementia; 37% for women with cerebrovascular disease (26% for cerebral infarction), 20% for women with skeletal diseases, 18% for women with dementia. CONCLUSIONS: The active life expectancy for women is longer than for men, by 3.7 years for the age of 65, by 2.3 years for 75 and by 0.5 years for 85. The dependent life expectancy for women is also longer than for men, by 3.2 years for the ages of 65 and 75 and by 2.6 years for 85. Thus, nursing prevention is an urgent issue, especially for women. The disorders requiring particularly long age-adjusted nursing time are carebrovascular disease (particularly cerebral infarction), dementia and skeletal disorders (particularly among women). PMID- 12087769 TI - [Factors related to under-utilization of in-home services under the long-term insurance system]. AB - PURPOSES: This study was conducted to examine factors associated with under utilization of in-home services covered by the long-term care insurance. Under utilization was measured in terms of the discrepancy rate between benefits limits and service costs for individuals. Based on the behavioral model, possible predictors were chosen; the level of nursing care as the need factor, living arrangements and income as enabling factors, and family caregiving consciousness as a predisposing factor. Both direct and interaction effects of those factors were examined. METHODS: 1,500 people were randomly selected from all people certified as needing long-term care in one ward, Tokyo to be interviewed. People categorized as "requires assistance" or "requires nursing care level 1" were interviewed directly. With people categorized as "level 2" to "level 5", their primary caregivers were interviewed. Information about in-home service utilization was collected from the insurer. RESULTS: People who were categorized as requiring a lower level of nursing care, lived with others, had a low income, or had high family caregiving consciousness demonstrated significantly greater under-utilization rate as compared with others. An interaction between living arrangements and the level of nursing was observed in this regard. While people who lived alone exhibited low a level of under-utilization rate without regard of the level of nursing care, the rate changed by those who lived with other. CONCLUSIONS: Under the new system, informal support might have a significant impact on under-utilization of in-home services. The responsibility for paying 10% of total care costs might be related to under-utilization by people with low income, though reduction of individual co-payments has been introduced for individuals in difficult financial conditions. PMID- 12087770 TI - [A survey of smoking behavior among students of Himeji-Dokkyo University]. AB - PURPOSE: In 2000, a survey of smoking behavior among students of Himeji-Dokkyo University was conducted in order to provide a basis for no smoking movements in the university. METHOD: Anonymous questionnaires were distributed to students and then gathered it. The analysis examined the relationships among sex and age and the smoking status (smoker, previous smoker, non smoker). RESULTS: Of the smokers, 44.0% started before the age of seventeen years. Non-smokers were more critical of smoking than smokers, with previous smokers occupying an intermediate position. No clear reasons were given for starting smoking although the behaviour of elder students was of importance. Smokers thought that it was desirable to reduce the smoking rate in Japan and that information should be available regarding effects on the body. A small proportion also were in favour of banning cigarette vending machines. CONCLUSIONS: Most answer to the question about starting his/her smoking were no positive reason. However, the effect of circumstances is important, because smoking of friends, senior students and family members had a strong influence. It was suggested that many smoking students know smoking is not good for their health, and would like to quit. PMID- 12087771 TI - [A survey on the dietary and health habits according to action styles for unbalanced diets among female university students]. AB - OBJECTS: A survey of woman's students, who had grown up in a time of change-over of Japanese customs regarding food, was conducted to investigate dietary habits, a focus on actions in response to with set needs, The aim was to obtain to measure prospective health maintenance and promotion. METHODS: The authors carried out a questionnaire for woman's students in 6 universities of Kanto area (a junior college and graduate courses were included) in years 1999 and 2000. RESULTS: In actions regarding the diet, 19.1% of the 815 students responded "I eat everything without fail", 17.7% "I eat almost everything irrespective of likes and dislikes", 24.5% "I strive to eat everything", 19.8% "I leave almost everything I dislike", and 18.5% "I don't eat what I dislike". The students who responded "I leave almost everything I dislike" and "I don't eat what I dislike" had low scores in the questionnaire for dietary and health habits, and they tended to like noodles and popular snacks, and also made frequent use of fast food restaurants. There was not a problem with the majority, but about 30% of the students were judged as confirming an unbalanced diet by this questionnaire. It was found that there were "gaps" and "bad habits" in the regular diet. Moreover, we recognized a tendency to prefer convenience and taste to nutritional balance. CONCLUSION: This simple questionnaire is suitable for rapid identification of unbalanced distory habits, and both "gaps" and "bad habits" in the regular system diet. It is the most important that we induce students who leave or don't eat what they dislike, to strive to eat everything, irrespective of likes and dislikes. PMID- 12087772 TI - [Construction and application of an information transfer system on prevalence of infectious disease for subscriber in a regional medical association]. AB - PURPOSE: To assess the utility of an e-mail information exchange system, for subscription by members of a regional medical association, focusing on surveillance of infectious disease. METHODS: Questionnaires regarding addition to the mailing list were first addressed to all 474 members of Yokkaichi regional medical association. After management for about three months, questionnaires focused on utility and application of the system were mailed out. RESULT: One hundred and seventy-three members (36.5%) responded in the questionnaire survey, then 73 of them (subsequently 75) actually subscribed. Forty-six subscribers (61.3%) responded to the second questionnaire, and 23 of them (56.5%) were in favor of 'weekly and monthly infectious disease reports transmitted from our research division', and 22 (47.8%) were in favor of 'e-mail exchanges of information'. CONCLUSION: These findings suggest that an electronic information transfer system is a useful medium for regional infectious disease efforts and contributions from primary care physicians. PMID- 12087773 TI - [Recommendations for making quality improvements in administration within local authorities]. AB - OBJECTIVE: To ascertain the methods and issues of industrial administration, that have been systematically chosen by the health and welfare administration in the Iwate Prefectural Government, in order to increase public customer satisfaction. METHODS: Combined use of TQM and marketing is thought to be the most effective way to increase public customer satisfaction with limited resources. This method secures a quality administrative service capable of corresponding to public needs, by improvement of processes and resources. Therefore we made use of both TQM and marketing. RESULTS: According to a customer satisfaction survey conducted by Iwate Prefectural Government in May 2001 (mail survey, sent to 234 people, response rate of 88.9%), public customer satisfaction had increased compared to the previous year in the following areas: provision for the elderly, the declining birthrate, disabled persons and universal design. Also, in a policy evaluation conducted during the same year, 207 services were revised and 30 were temporarily closed, abolished or reduced, 20 were merged, and 23 were expanded. CONCLUSIONS: We cannot sufficiently evaluate the effectiveness of the methods yet because they were only just introduced. However, development of the plan and policy evaluation appear to have been quite effective. From now on we will continue to improve our methods, especially to increase the quality of our administrative resources. PMID- 12087774 TI - [The plant origins of herbal medicines and their quality evaluation]. AB - The caulis (stem and leaf) of Trachelospermum jasminoides (Lindl.) Lem. (Apocynaceae) is listed as the plant origin of Luoshiteng in the Chinese Pharmacopeia. However, preparations from the caulis of Ficus pumila L. (Moraceae) or Psychotria serpens L. (Rubiaceae) are distributed on the Chinese market. The fruit of Forsythia suspensa Vahl (Oleaceae) is listed as the plant origin of Forsythia Fruit in the Chinese Pharmacopeia, although the fruits of two Forsythia species, F. suspensa and F. viridissima Lindley, are listed as the plant origins in the Japanese Pharmacopeia, and fruits of three Forsythia species, F. viridissima, F. koreana Nakai, and F. suspensa, are listed in the Korean Pharmacopeia. The whole plant of Plantago asiatica L. (Plantaginaceae) is listed as the plant origin of Plantago Herb in the Japanese Phamacopeia, but the whole plants of two Plantago species, P. asiatica and P. depressa Wild, are listed as the plant origins in the Chinese Pharmacopeia. The leaves of two Plantago species, P. lanceolata L. and P. major L., are distributed as Plantain on the European market. Each of these herbal medicines is reviewed based on the differences in plant origins and their quality evaluation from the viewpoints of the morphological properties, chemical components, and biological activities, respectively. PMID- 12087775 TI - [Case report summary browsing system for education of pharmaceutical students through the Internet]. AB - We have developed a patient case database at Gunma University Hospital. The transmission of the data contained in this database via the Internet is protected by SSH encryption technology. The database may also function as an education tool for medical and pharmaceutical students who can access this system through the Internet using a special browser system that we have also developed. In a survey questionnaire relating to our system conducted among graduate students, most responded positively to its usefulness as an opportunity for exposure to actual clinical practice. Because this system can introduce students to realistic and daily practices at hospitals, it might have a great impact on Japanese pharmaceutical education in universities or pharmaceutical colleges where early exposure to actual hospital practices is not appropriately included in the curriculum. In addition, our system will function to establish a mutual feedback system between hospitals and basic research institutions such as pharmaceutical universities. PMID- 12087776 TI - [A checking system for injectable anticancer drugs using each patient's own data and its evaluation]. AB - A checking system was developed for risk management of injectable anticancer drug use. Because the dosage and administration of injectable anticancer drugs vary with each patient, checking the prescription and aseptic i.v. admixing can be markedly complicated and time-consuming for pharmacists. The system we investigated in this study provided real-time checking of dosage, drip infusion rate, dosing periods, and dosing-free periods. The prescription parameters for this check included height, weight, body surface area, the medical history of each patient, the patient's ICD10 code' and disease indication from the package insert. Moreover, when preparing for aseptic i.v. admixing, the liquid volume after the anticancer drug has been mixed with other injectable drugs is calculated automatically. The time limits for stability after mixing and clinical laboratory test results are listed on the Work Sheet. As a result, 24 medication errors were prevented in the first 6 months of use of this system. These consisted of over- and under-doses, excessive dosing periods, insufficient dosing free periods' etc. The time required for preparation of aseptic i.v. admixing decreased by 73% after introducing this system. Because clinical laboratory test results were referenced beforehand, patients to whom the drug should not be administered were recognized in advance. Thus 13 such cases were identified before i.v. admixing. Therefore this system may be useful in terms of rational anticancer drug use and risk management. PMID- 12087777 TI - Herbal medicine and false-positive results on lymphocyte transformation test. AB - In vitro mitogenic activity of 16 herbs and 3 Kampo (herbal medicine) formulae have been reported in experimental studies. It is not known how many herbs and Kampo formulae in total have mitogenic activity. Lymphocyte transformation test (LTT) is generally utilized to diagnose drug-induced liver injury. In LTT, mitogenic activity is assessed by measuring 3H-thymidine incorporation. The objective of the present study was to determine which herbs and which Kampo formulae caused false-positivity on LTT. We examined 2496 summaries of all admission records from 1979 to 1999 in our department. We selected patients in whom liver injuries were diagnosed as definitely unrelated to Kampo medication. In these patients, LTT was performed for some herbs contained in the suspect Kampo medicines, resulting in positive LTT for 17 herbs: Evodiae Fructus (Goshuyu), Zizyphi Fructus (Taiso), Ginseng Radix (Ninjin), Zingiberis Rhizoma (Shokyo), Hoelen (Bukuryo), Aconiti Tuber (Bushi), Angelicae Radix (Toki), Cnidii Rhizoma (Senkyu), Rehmanniae Radix (Jio), Ephedrae Herba (Mao), Anemarrhenae Rhizoma (Chimo), Cinnamomi Cortex (Keihi), Bupleuri Radix (Saiko), Artemisiae Capillari Spica (Inchinko), Persicae Semen (Tonin), Moutan Cortex (Botanpi) and Paeoniae Radix (Shakuyaku). These results were considered false-positive, because the results were observed in the "definitely unrelated" patients. Mitogenic activity inherent to some herbs and Kampo formulae may sometimes cause false positivity on LTT in clinical situations. These examples suggest that LTT for Kampo formulae may be unreliable as a diagnostic method for drug-induced liver injury. PMID- 12087778 TI - Rhubarb use in patients treated with Kampo medicines--a risk for gastric cancer? AB - In vitro mutagenic effects have been reported for ingredients contained in rhubarb. Therefore, rhubarb (Rhei Rhizoma) as an anthranoid laxative could be associated with a risk of developing gastric cancer as well as colorectal cancer. We are not aware of any reports that have examined the relationship between the use of rhubarb and the development of gastric cancer. During the period between 1979 and 1999, we treated 14,616 patients using various Kampo medicines, which sometimes contained rhubarb. In the present study, we determined whether patients, diagnosed with gastric cancer during the period between 1979 and 1999, had been administered rhubarb before the development of gastric cancer. Among the 10 enrolled patients, only 2 patients had been administered rhubarb before the development of gastric carcinoma. The other 8 patients had never received rhubarb before the development of gastric carcinoma. Rhubarb use may have little connection with the development of gastric cancer in practice, even if some ingredients in rhubarb have shown carcinogenic activity in experimental studies. PMID- 12087779 TI - [Relation of bile hydrogen peroxide level and liver super oxide dismutase activity in selenium-deficient rats]. AB - The relationship of hydrogen peroxide (H2O2) levels in bile with liver SOD and GSH-Px activity in selenium (Se)-deficient rats is discussed. Normal rats and 7 groups of rats fed a Se-deficient diet with different feeding periods were examined. H2O2 levels in bile were measured using the spin-trapping method with electron spin resonance (ESR). Bile H2O2 levels in the initial stage (20-60 min from start of the cannulation) of measurement were increased depending on the length of the feeding period with the Se-deficient diet and absence of Se. Bile H2O2 levels in the later stage (60-120 min) of measurement first increased with the length of feeding with the Se-deficient diet and then decreased with longer feeding periods. Bile H2O2 levels immediately after the operation were relatively low in almost all cases. The operation may result in oxidative stress to generate H2O2. Liver GSH-Px activity decreased depending on the length of the feeding period with the Se-deficient diet and existence of Se. Liver SOD activity increased in Se-deficient groups. It is suggested that the H2O2 levels in bile are related to decreased GSH-Px activity, SOD activity, and also the oxidative stress caused by surgery. Therefore the H2O2 levels in bile can be used as an index of sensitivity to oxidative stress. Although severe oxidative stress may decrease SOD activity, Se deficiency can induce liver SOD activity. PMID- 12087780 TI - [Interaction between acyclic phenol--formaldehyde oligomers and quaternary ammonium ions]. AB - The interaction between acyclic phenol-formaldehyde oligomers (1) and quaternary ammonium ions (5) was investigated by 1H-NMR spectroscopy. From the induced chemical shift change of 5 in the presence of 1, the cation-pi interaction between the N(+)-(CH3)3 moiety of 5 and the pi-base of 1 occurs during the formation of the complex. Another important observation is that the OH proton signals of 1 are considerably broadened upon addition of 5, indicating that hydrogen bonding occurs between exchangeable protons of 1 and iodide anion of 5. These interactions play an important role in the binding of 5. PMID- 12087781 TI - [Pilot study of the practical use of Internet in medical services: the questionnaire for pharmacists in Japan]. AB - We performed a questionnaire survey to 260 pharmacists as a pilot study, in order to know the actual application of IN to medical services in Japan. The response rate of our questionnaire was 40% (105 persons), and 91% of the answers were from pharmacists who works in hospitals, community pharmacies, or clinics. According to the results, 90 of the 105 pharmacists had had some experience of using IN, and 68% of whom (58% of respondents) use IN daily as a means of problem solution on their works. IN was probably used as a relatively reliable information source something like text-books in such cases as acquiring the medical information for patients or other medical staffs. In addition, IN may have been esteemed its facilities and the informational usefulness and reliability. However, since this survey is just a pilot trial, the result dose not necessarily reflect a general situation around IT use. Consequently, this study result could be need by still further research. PMID- 12087782 TI - National Kidney Foundation Guidelines for chronic kidney disease: evaluation, classification, and stratification. AB - Chronic kidney disease is becoming more prevalent in the United States. The National Kidney Foundation has recently published a new set of guidelines to assist clinicians in providing earlier detection and treatment of kidney disease to minimize the progression to end-stage renal disease. As approximately 11% of the adult population has some degree of kidney disease, this new CPG should be applicable in many settings. PMID- 12087783 TI - Implementing a smoking cessation program for pregnant women based on current clinical practice guidelines. AB - PURPOSE: To describe the U.S. Department of Health and Human Services clinical practice guideline for treating tobacco use and dependence and demonstrate how the guideline was utilized in a pilot program for a small sample of pregnant women (n = 20) to help them decrease smoking. DATA SOURCES: A convenience sample of 20 pregnant women was recruited from a health maintenance organization at their initial prenatal contact either by telephone or in person. A comparison group of pregnant women (n = 28) was used for analysis of outcomes. CONCLUSIONS: Clinical results showed better outcomes for women in the pilot program when compared to a similar group who did not participate in the program. There was a statistically significant difference between the two groups in average number of cigarettes smoked per day at delivery and two weeks after delivery with pilot program participants reporting less smoking (p < .05). Women in both groups showed a pattern of returning to smoking after delivery of the baby. IMPLICATIONS FOR PRACTICE: Although a few tobacco users achieve permanent abstinence in first or second attempts, the majority continue to use tobacco for many years and typically cycle through many lapse and relapses before permanent abstinence. Ambulatory care systems need to be developed and funded to treat tobacco use and dependence over the life span. Recognition of the chronic nature of the problem and development of long term care delivery systems are needed to assist clients to achieve goals of permanent abstinence and better personal and family health. This cycle of lapse and relapse before permanent abstinence is typical and demonstrates the chronic nature of tobacco use and dependence and the need for long term follow-up. PMID- 12087784 TI - Assisted living facilities as a site for NP practice. AB - PURPOSE: To describe what an assisted living facility (ALF) is who resides in ALFs, and the benefits of nurse practitioner (NP) practice in an ALF for the client, facility, NP, and practice. DATA SOURCES: Selected scientific literature, legal and regulatory guidelines, consumer information and guidelines, Internet sites, and the authors' experiences. CONCLUSIONS: Assisted living facilities are nonmedical residential settings grounded in a philosophy of providing residents with choice, autonomy, privacy, independence, dignity, and respect in a housing environment as homelike as possible. Nationally, ALFs vary considerably by size, services provided, and regulations imposed by states. IMPLICATIONS FOR PRACTICE: Assisted living facilities are now a popular choice for individuals who desire assistance with selected activities of daily living (ADL) while maintaining maximum independence. They are becoming an excellent site for the provision of in home primary care in order to facilitate aging in place. PMID- 12087785 TI - Nuts and bolts of NP practice in assisted living facilities. AB - PURPOSE: To describe the practical issues associated with setting up a nurse practitioner (NP) practice in an assisted living facility (ALF), including record keeping, maintaining communication, and scheduling and coordination issues. DATA SOURCES: Selected scientific literature, legal and regulatory guidelines, and the authors' experiences. CONCLUSIONS: Initiating a NP practice in an ALF can be a rewarding experience, but it will not be without its own unique challenges. Procedures that maintain excellent communication between the NP, the home practice site, the resident and family, the ALF facility, and other services required by the needs of the resident take time and effort to establish but are worth the extra effort. IMPLICATIONS FOR PRACTICE: Nurse practitioners who establish a practice in an ALF need to be familiar with state regulations, facility procedures and culture, and the needs of elders who are aging in place and integrate all that into their office practice sites in order to be successful. PMID- 12087786 TI - Chronic cough: evaluation and management. AB - PURPOSE: To inform the nurse practitioner of the common causes of chronic cough, to educate how each cause can be identified, and to suggest treatment options for the most common causes. DATA SOURCE: Selected research and review articles from professional journals. CONCLUSION: Chronic cough is a common presenting complaint in the primary care setting. Although at first the symptom may seem simple, the etiology, diagnosis, and treatment of chronic cough can be challenging. Furthermore, without proper identification of the cause, effective treatment most likely will not be obtained. IMPLICATIONS FOR PRACTICE: By familiarizing oneself with this topic, nurse practitioners will feel confident in treating chronic cough. PMID- 12087787 TI - Bulimia nervosa: knowledge, awareness, and skill levels among advanced practice nurses. AB - PURPOSE: To examine the knowledge, awareness, and self-perceived skill levels regarding bulimia nervosa (BN) among advance practice nurses (APNs) caring for young female patients. DATA SOURCES: Two hundred APNs of various nurse practitioner (NP) specialties were surveyed with a mailed questionnaire. One hundred and twenty-one (61%) surveys were returned and analyzed. CONCLUSIONS: Results demonstrated that APNs are knowledgeable and aware about BN. There were several statistically significant relationships among awareness and self perceived skill levels. Pediatric NPs were found to be the most aware of BN in their young female patients. IMPLICATIONS FOR PRACTICE: The incidence of eating disorders is on the rise. Bulimia nervosa, a common secretive illness, poses a significant challenge for APNs potentially leading to missed diagnosis. There is a paucity of research exploring this issue. Study findings provide evidence to enhance curricula on eating disorders in academic and continuing education programs and guidance for future research. PMID- 12087788 TI - Doctorial dissertations on nurse practitioners: 1970-2000. AB - PURPOSE: To understand more about the nurse practitioner (NP) profession, a study was undertaken of the dissertation literature. DATA SOURCES: A search of Dissertation Abstracts Online yielded 134 dissertations that included NPs in the title or abstract and were awarded between 1970 and 2000. CONCLUSIONS: Most of the degrees are either PhD (61%) or EdD (27%). State-based universities produced the most degrees (70%) and six universities accounted for one-fourth of the degrees. The topics of NP-focused dissertations are wide ranging and include history, medical economics and managed care, NP education and professional role, and clinical and behavioral research. IMPLICATIONS FOR PRACTICE: This dataset provides an important source of information for research on the NP profession. PMID- 12087789 TI - A prostate cancer primer. AB - Prostate cancer is the second most commonly diagnosed malignancy in men and remains the second leading cause of cancer deaths among this population. Early screening and diagnosis, advances in preventive techniques, and existing cutting edge treatments are part of a multidimensional approach in an ongoing battle to improve the health of millions of men. This primer was designed to familiarize nurses with basic facts about prostate cancer and its management. The epidemiology of prostate cancer and its risk factors, clinical manifestations, screening and early diagnosis, treatment strategies, and implications for urologic nurses are presented. PMID- 12087790 TI - Rules to follow before advising patients on dietary supplements/alternative medicine: Part II. AB - Five rules for health care professionals to follow when patients inquire about complementary/alternative medicine are outlined. Suggestions include having health care professionals direct patients toward more objective medical articles such as reviews and editorials and to encourage patients to bring various advertisements and claims made by certain companies to the clinic so that health care professionals can analyze their overall merit. The Food and Drug Administration should be contacted by any health professional who has observed a negative or positive reaction with any dietary supplement. Health care professionals should read several alternative magazines and journals to better understand what patients are reading. Finally, patients should be assured that data gathering on some of these alternatives are being collected at a rapid pace that should promote a better understanding of the benefits and detriments of certain supplements. PMID- 12087791 TI - Characteristics of North American urodynamic centers and clinicians. AB - Centers performing urodynamic testing in North America were surveyed to identify both who was performing urodynamics and what techniques were employed during specific tests. Urodynamic laboratories tend to have one or more dedicated urodynamic units, capable of measuring multiple pressures, uroflowmetry, and sphincter electromyography. Testing is typically completed by a nurse, physician, physician's assistant, or technically skilled clinician with extensive clinical experience. However, few of these clinicians report formalized education in focusing specifically on urodynamic testing techniques. PMID- 12087792 TI - Nursing research and continence care. AB - Despite 50 years of attention to continence care and pivotal nursing research in the field, incontinence continues to challenge health care professionals and consumers alike. The role of nursing research in continence is traced from the early influences of Florence Nightingale to today's clinical practice guidelines and the influence of major nursing research centers. PMID- 12087793 TI - The senses interrupted. PMID- 12087794 TI - The art of raising the bar. PMID- 12087795 TI - Getting ready for certification: urolithiasis. PMID- 12087796 TI - Xigris: reducing mortality in adult patients with severe sepsis. PMID- 12087797 TI - [The new system of compensation of occupational accidents and diseases: features and details of the reform introducing the concept of biological damage]. AB - The compensation system concerning accidents at work and occupational diseases has been substantially modified by the Accidents at Work and Occupational Diseases Insurance (Amendment) Regulations 2000 which introduced compensation for biological damage. This kind of impairment is already a well known legal and medical concept in civil liability. Therefore, while in the past the generic working capacity was taken into account in order to obtain the impairment percentage, this kind of evaluation is now based on the damage to biological and relational capacities of the person. Thus, while the medical premise evaluated in percentage, i.e. the biological damage, unites INAIL's system with the ordinary system of civil liability, the economic evaluation highlights difference between the two compensation systems. The system as modified by the above mentioned Regulations provides, along with compensation for biological damage, compensation for financial losses caused by psycho-physical impairment, i.e. impairment of working capacity. The legal instruments provided by Parliament to implement the two kinds of compensation (biological damage and financial losses) were passed by the Secretary of State for Employment with Approval of Tables of Impairments, Biological Damage and Coefficients Regulations 2001. Three tables have been approved: the first with impairments in order to have a percentage evaluation of the biological damage; the second with the coefficients to obtain the percentage of impairment to working capacity; the third for the economic conversion of the impairment percentage evaluated with the first two tables. The paper goes over the framework of the new protection system, analyses the instruments for its implementation, i.e. the tables, and explains methods for their application. PMID- 12087799 TI - [Are we physicians or corporals?]. PMID- 12087798 TI - [Health surveillance of workers formerly exposed to occupational cancerogenic substances: various operative views]. PMID- 12087800 TI - [Angioneurosis caused by vibrating devices: the first contributions to the literature by Italian authors]. PMID- 12087801 TI - [Prostate diseases of occupational origin]. AB - BACKGROUND: Prostatitis is a poorly defined group of syndromes with multiple causes. Chronic prostatitis may be non-bacterial and due to intrapelvic venous congestion. If the causes persist and adequate treatment is not given, the congestive syndrome may become chronic and interfere with fertility with severe biological damage. OBJECTIVES: Little is known in the field of occupational medicine (as regards clinical and pathogenic aspects) and, on the other hand, little is known by urologists (as far as the aetiological aspects are concerned), the prostatitis-like syndrome due to intrapelvic congestion has been defined in recent studies as non-bacterial prostatitis or prostatodynia, but we prefer to call it "prostatosis". The results of a close cooperation between urologists and occupational physicians are reported. METHODS: Patients with non-bacterial chronic prostatosis were evaluated from the urological and occupational point of view and all the etiological factors of both occupational and non occupational origin were considered. When occupational factors are a conditio sine qua non prostatosis is considered an occupational disease. RESULTS: Two cases of "occupational prostatosis" are described where driving vehicles and a sedentary employment played the most important etiological part. Many other similar cases were observed. CONCLUSIONS: Close cooperation between urologists and occupational physicians makes it possible to complete clinical diagnosis with a careful evaluation of all the factors of both occupational and non-occupational origin and allows the identification of those cases that must be defined as occupational diseases. Prostatosis due to venous congestion deserves the attention of occupational physicians since the distinction between occupational and non occupational origin must be found both in individual cases and in groups of workers subject to the same factors. Moreover, the present state of knowledge is enough to take preventive measures aimed at reducing the frequency of new cases and avoiding the deterioration of existing cases. It is also possible that some cases of pseudo-cystitis in women might be the result of intrapelvic venous congestion of occupational origin. Anatomical and physiological non-occupational factors as well as certain habits of life style can favor intrapelvic venous congestion, producing conditions of hypersusceptibility to occupational factors and could sometimes cause the disease even in the absence of causes connected to work. PMID- 12087802 TI - [RISKOFDERM: European research project for assessment of occupational skin exposure to chemicals]. AB - OBJECTIVES: RISKOFDERM is a research project whose aim is to develop instruments to assess and manage occupational dermal exposure to chemical substances. METHODS: The research, funded by the European Commission, involves 15 Institutes from 10 member countries; it is a continuation of the Dermal Exposure Network experience and consists of four interrelated parts. The first phase (Qualitative survey) assumed that dermal exposure can be extrapolated from one compound to another when it is task-based: therefore six Dermal Exposure Operation units (DEOu) were defined that lead back to the variety of occupational dermal exposure conditions and an extensive Questionnaire was developed for on-site surveys to perform standard observations in selected working situations (scenarios). RESULTS: The Italian group, participating in the research, obtained a set of observations relating to two "scenarios" in different working sectors: asphalt, ceramic and pottery workers, spectacle decorators and paint production: the aim was to verify the validity of the methodology in assessing the risk of percutaneous absorption, time, frequency and extension of skin exposure. CONCLUSIONS: From the observations made it was shown that the perception of risk was poor; it is necessary to rationalise work organization, and train and inform the employees on the correct use of personal protection devices. PMID- 12087803 TI - [Rhizarthrosis of the thumb in ironing workers]. AB - BACKGROUND: Rhizarthrosis of the hand secondary to job-related static-dynamic stress is a rare occupational condition characterised by secondary osteoarthrosis. OBJECTIVES: We report 5 clinical cases of female ironers employed in small-scale laundries, who after more than 10 years on the job developed pain in the right thumb. Disease onset may have been favoured by the continuous functional stress exerted on the joint. METHODS: Clinical examination, hand X rays and ultrasonography of the trapezio-metacarpal joint led to the diagnosis of rhizarthrosis. The electromyographic and biomechanical analysis of work tasks was conducted by simulating normal ironing activity. RESULTS: Patient history and electromyographic and biomechanical data showed a close causal relationship between age, job and diagnosis. CONCLUSIONS: In the case of workers over 50 years of age whose jobs have the characteristics described here, Finkelstein's test should be performed on occasion of occupational health surveillance to screen for the arising of the disease. PMID- 12087804 TI - [Prevalence of occupational allergy to laboratory animals in two towns of northern and central Italy]. AB - BACKGROUND: Laboratory animal allergy (LAA) is a well known occupational hazard for workers employed in biological and medical research institutes, but few Italian data on this disease exist. OBJECTIVES: The aim of our study was to evaluate the prevalence of LAA in 45 workers in Trieste (NE-Italy) and in 80 workers in Perugia (Central Italy) and to assess factors predisposing to sensitisation among subjects occupationally exposed to animals. METHODS: All subjects underwent a physical examination and responded to a standardized questionnaire for the evaluation of allergic respiratory symptoms and exposure data. Skin prick tests with common allergens and with hair extract from laboratory animals were performed and specific IgE was measured. Atopy was defined as positive skin prick test to common allergens. RESULTS: There were 60% atopic subjects in Trieste and 55% in Perugia and sensitisation to laboratory animal hair was found in 24.4% subjects in Trieste and in 35% in Perugia. The prevalence rates of LLA were respectively 11.1% and 11.2%; 2.3% and 3.7% complained of asthma while 8.9% and 7.5% complained of rhinitis. The resulting symptoms were significantly related to skin prick tests that were positive to laboratory animal hair (odds ratio (OR) = 7.64; 1.83-44.5), to skin prick test positivity to common inhalant allergens (OR = 5.29; 1.09-50.2), to common allergic symptoms (OR = 3.95; 1.05-18.2) and to exposure time exceeding 5 hours per day (OR = 5.45; 1.31-22.0). CONCLUSION: The role of atopy and of exposure time in causing LLA was confirmed and the need of prevention measures to reduce exposure in people at risk was discussed. PMID- 12087806 TI - [Therapeutic immunosuppression: knowledge improvement]. PMID- 12087805 TI - [Cohort study of mortality among leather tanners in the Lower Valdarno area]. AB - BACKGROUND: Epidemiological studies of tanners have shown increased risk for a number of cancer sites, namely: lung, bladder, kidney and urinary organs as well as stomach, intestine, pancreas, nose and nasal cavities, together with leukemias and soft tissue sarcomas. OBJECTIVE: To study cause specific mortality of leather tanners in Tuscany (Valdarno Inferiore area). METHODS: The cohort included 4874 workers (4150 males and 724 females) employed in 92 tanneries operating in 1996 (Valdarno Inferiore Tanneries Census) which were also operating on 31-12-1970. Ascertainment of vital status was completed for all individuals on 31-12-1998 (end of follow-up), and the cause of death was known for all deceased subjects. Demographic and work history data were obtained from factory payrolls. Regional mortality rates were used for comparison to calculate SMR (Standardised Mortality Ratio) and 90% Confidence Intervals (CI). In addition to the overall cohort analysis, for men only separate analyses were completed for finishers, chrome tanners and vegetable tanners. RESULTS: The study showed an increased mortality from lung cancer among finishers, Standardised Mortality Ratio (SMR) 145, 19 observed (obs) (90% Confidence Intervals, 90% CI 95-212), from bladder cancer in the overall cohort (SMR 134, 9 obs, 90% CI 70-233) and among finishers (SMR 125, 2 obs, 90% CI 22-393) and from pancreatic cancer among finishers (SMR 120, 2 obs, 90% CI 21-379). Mortality from lymphoemopoietic cancer is above expected, and the increase is mainly due to myeloid leukaemia, both in males (SMR 208, 5 obs, 90% CI 82-437) and females (SMR 599, 2 obs, 90% CI 106-1887). No deaths from soft tissue sarcoma were observed. A new finding of the study was the increased mortality from cancer of the endocrine glands (SMR 566, 4 obs, 90% CI 194-1297), psychiatric disorders (SMR 195, 6 obs, 90% CI 85-385) and blood diseases (SMR 329, 4 obs, IC 90% 112-752). CONCLUSIONS: The observations of increased lung cancer mortality among finishers, of bladder cancer in the overall cohort and among finishers, as well as an increase in pancreatic cancer among the latter, confirm previous epidemiological findings among tanners. The increase in myeloid leukemia mortality for both males and females, and the absence of deaths from cancer of the connective tissue, which includes soft tissue sarcomas, are worthy of note. The results should be valued with caution, given the small number of cases and the novelty of some observations. PMID- 12087807 TI - [Function of the CLC chloride channels and their implication in human pathology]. AB - To date, nine chloride channels belonging to the family of CLC chloride channels have been identified. They are localized either in plasma membranes or in intracellular vesicles (endosomes or lysosomes) and can have an ubiquitus or a more restrained tissue distribution. Recent studies on ClC-K1, ClC-2, ClC-3, ClC 5 and ClC-7 knockout mice and the identification of human inherited diseases caused by mutations of some of these chloride channels (myotonia congenita for ClC-1, Bartter disease for ClC-Kb, Dent's disease for ClC-5 and osteopetrose for ClC-7) have provided lines of direct evidence of the physiological relevance and importance of these chloride channels in the transport of chloride and in the endocytosis and transcytosis of proteins in specialized cells from the kidney and other tissues. PMID- 12087808 TI - [Nephrotoxicity of amphotericin B]. AB - Amphotericin B is widely used for severe life threatening fungal infections. Its use is limited by a dose-dependent nephrotoxicity manifested by a reduction in glomerular filtration rate and tubular dysfunction. An elevated creatinine associated with amphotericin B is not only a marker for renal dysfunction but is also linked to a substantial risk for the use of hemodialysis and a higher mortality rate; therefore amphotericin B nephrotoxicity is not a benign complication and its prevention is essential. Several manipulations have been proposed to try and minimize amphotericin B induced nephrotoxicity. Systematic hydration is crucial to minimize amphotericin B. Mannitol or intralipids administration were once suggested as protective based on anecdotal observational reports. Small prospective and randomized trials, however did not support a protective effect. Three new formulations have been developed in an attempts to improse both efficacy and tolerability: amphotericin B in lipid complex (ABLC, Abelcet). Colloidal dispersion (ABCD, Amphotec and amphotericin B liposome (Ambisome). Three prospectives randomized studies have clearly shown that Ambisome is less nephrotoxic than amphotericin B. Unfortunately the only randomized trial comparing Abelcet with amphotericin B is an open-label treatment of invasive candidiasis which was presented 5 years ago but never published as a full paper. Furthermore in a recent multicenter double-blind study it has been shown that Ambisome has a better safety profile than Abelcet with less chills/rigors and less nephrotocixity. PMID- 12087809 TI - [Evolution of medical practice in the care of anemia amd the use of erythropoietin in chronic renal insufficiency hemodialysis in the past six years]. AB - In order to analyze changes in anemia management in the hemodialysis patients, a retrospective survey was performed in 1999 on a representative French sample of patients in hemodialysis treated by EPO since 1992, 1994, 1996, and 1998 respectively. 368 patients were enrolled in 97 centres. At treatment initiation, some patients characteristics have significantly changed: the percentage of type 2 diabetic patients has increased marquedly from 4.5% in 1992 to 22.1% in 1998. The mean time lag between dialysis and EPO treatment initiation has decreased from 3.3 years to 0.9 on the 6-year period. Lastly, the mean hemoglobin level has increased from 73.7 to 78.5 g/l. No significant evolution in the EPO protocol was observed at the initiation as well as in the maintenance phase. An average percentage of 33.7% of patients were treated in i.v. route at initiation with a mean posology of 105.2 IU/kg (respectively 91.4 in s.c.). In the first year of follow-up, the overall mean posology was 84 IU/kg. In conclusion, this survey suggests that anemia management has been optimized over the last 6 years follow up, including an earlier initiation of EPO treatment and a higher target of hemoglobin while EPO needs were reduced in the same time. PMID- 12087810 TI - [Colonic necrosis caused by sodium polystyrene (kayexalate) in hemodialysis: myth or reality? Two case reports]. AB - Among gastro-intestinal complications reported in uremic or transplanted patients, colonic necrosis due to Kayexalate and sorbitol is not so unusual for english authors. This iatrogenic event is probably underestimated by physicians and pathologists. In France, neither literature nor pharmacovigilance refer this complication but Vidal dictionary advises against such association. We thought of interest to report 2 representative observations and, after reviewing literature, to discuss diagnostics circumstances and etiopathogenic hypothesis and then to give some advices. PMID- 12087811 TI - [Tuberculosis in hemodialysis patients in Tunisia]. AB - BACKGROUND: Because of the immunity depression, patients with chronic renal failure undergoing haemodialysis are at increased risk for developing infections, Mycobacterium tuberculosis (MTb) in particular. OBJECTIVE: To evaluate, in a prospective longitudinal study over 36 months period, the frequency of Tb among these patients and underline the diagnostic difficulties. PATIENTS AND METHODS: Sixty dialysis patients were interested. The Tb assessment comprised: a questionnaire, a meticulous clinical examination, a chest X-ray, a tuberculin skin testing, as well as MT bacilli screening in biological fluids. RESULTS: Tb was seen in six among dialysis patients(10%), this rate represents 15 times the general population tuberculosis incidence in our country (23/100,000). Tb has occurred early compared to the beginning of the haemodialysis. The Tb localizations were: 4 cases of extra-pulmonary tuberculosis (3 peritoneal and one ganglionic) and only 2 cases of pleuro-pulmonary localization. The positive diagnosis represents a real problem: no bacteriological confirmation in all patients and only in one, the histological diagnosis was obtained. CONCLUSION: There was a high rate of tuberculosis in our study. The prognosis appears to be closely related to therapeutic precocity, thus specific chemotherapy started sometimes without diagnostic confirmation. PMID- 12087812 TI - Imiquimod 5% cream: a topical immune response modifier. PMID- 12087813 TI - Imiquimod in the treatment of actinic keratoses and other intraepithelial neoplasms. PMID- 12087814 TI - Imiquimod 5% cream: a new treatment option for basal cell carcinoma. PMID- 12087815 TI - Imiquimod stimulates innate and cell mediated immunity which controls virus infections and tumors. PMID- 12087817 TI - [The Ufa research-scientific institute of industrial medicine and human ecology: results and prospects]. PMID- 12087816 TI - Imiquimod: a new immune response modifier for the treatment of external genital warts and other diseases in dermatology. PMID- 12087818 TI - [Evaluation of health status among workers in the oil mining industry and problems of primary prophylaxis]. AB - Thorough medical examination covered 1,400 individuals who work with oil and gas extraction devices, are aged 20 to 58 and have length of service 5 to 30 years. Findings are high prevalence of locomotory disorders, cardiovascular and gastrointestinal diseases, signs of ear exposure to noise. The authors justified complex of primary prophylactic measures including work optimization, meticulous occupational selection, qualified medical control and sanative measures. PMID- 12087819 TI - [Prophylactic efficiency of tietasol and oxymethyluracyl against chlorphenols exposure in experimental and real conditions]. AB - The authors studied protective efficiency of tietasol and oxymethyluracyl in experiments with animals exposed to 2,4-dichlorphenol and in real chlorphenol production. According to the results, oxymethyluracyl could be recommended to increase resistance against chemicals in occupational conditions. PMID- 12087820 TI - [Methodologies as applied to the organization of socio-industrial monitoring in regions with developed petroleum chemical industry and oil refineries]. AB - The authors tackle a problem of social and hygienic monitoring organization in territories with developed oil chemistry and oil processing industries. Analysis of longstanding observations over environmental objects and public health state serves to justify priority criteria to be included into social and hygienic monitoring system. PMID- 12087821 TI - [Psychophysiological features of fatigue in police officers during service]. AB - Peculiar occupational factor in police officers service is intensive psycho motional and physical strain leading to performance decrease caused by fatigue, to high transitory disablement morbidity. Studies enabled to specify and put into practice effective medical and prophylactic measures, to increase performance in criminal police officers, duty officers and road police staffers. PMID- 12087822 TI - [Logic and evaluation of industrial ecological risks in industrial cities in Russia]. AB - Industrial workers play a significant role in formation of ecologic technogenic risks in industrial cities of Russia as are subjected to technogenic load considerably higher than that of the general population and cover over a half of employable and pensionary urban residents. PMID- 12087823 TI - [Specific oncological skin lesions in glass fibre production]. AB - During 1986-1999 a total of 49 patients having occupational skin diseases are followed up in Occupational Diseases Center of Bashkortostan Republic. These diseases tend to increase, especially in glass fibre production. As occupational hyperkeratosis and skin cancer increase in workers engaged into glass fibre production, such industry could be considered as oncologically risky. PMID- 12087824 TI - [Occupational and ecologic components of regional industrial health risk in population exposed to mercury]. AB - Having total ecologic load considerably higher than that among general population, industrial workers form a priority risk group in ecologically unfavorable region characterized by environmental pollution with variable toxic chemicals. Nearly 70% of risk in the industrial workers are due to occupational factors. However, higher mercury content of biologic materials in the general population, being slightly lower than that in the industrial workers, could indicate possible specific conditions caused by mercury exposure in some residents of the settlement under study. PMID- 12087825 TI - [Intestinal micro-biocenosis in workers engaged in oil extraction]. AB - Nearly one third of workers engaged into oil extraction suffer from gastrointestinal disorders. Stationary medical examination revealed over 70% diseased subjects among which 98% of cases demonstrated disorders of colic microbiocenose (absent or depressed normal microflora, growing pathogens, deficient Bifidum bacteria in 63% of the examinees). These facts could be caused by environmental (including occupational) hazards resulting in decompensation of adaptive mechanisms. PMID- 12087826 TI - [Treating somatic dysfunctions in the pathogenesis of vibration disease]. AB - The authors revealed numerous somatic dysfunctions in vibration disease. The dysfunctions appeared to correlate with the vibration type and to play a role in pathogenesis of pain, tunnel and sensory polyneuropathy syndromes. Manual therapy proved effective in correction of somatic dysfunctions in vibration disease. PMID- 12087827 TI - [Chronic and acute stress in financial activities]. PMID- 12087828 TI - [Changes in occupational morbidity among workers of petroleum, oil-processing and petroleum chemical industries over 40 years]. AB - Analysis covered 3784 cases of occupational diseases diagnosed first in Research Institute for Industrial Medicine and Human Ecology in Ufa from 1960 till 2000. Locomotory and peripheral nervous system disorders appeared to predominate in structure of occupational morbidity. The authors failed to reveal direct correlation between occupational morbidity level and work conditions jeopardy class. PMID- 12087829 TI - A matter of trust. PMID- 12087830 TI - Missing cash--now what? PMID- 12087831 TI - What to do when government agents come calling. PMID- 12087832 TI - Leadership: the young dentist's secret to success. PMID- 12087833 TI - Early defibrillation--the key to survival. PMID- 12087834 TI - Dr. John Buchheister: "Let's make the MDA an even better association!" Interviewed by Michael G. Maihofer. PMID- 12087835 TI - The disappearing tooth: report of a case. AB - BACKGROUND: The possibility of a patient ingesting or aspirating foreign bodies exists in dentistry. Measures are available to minimize this risks. CASE DESCRIPTION: The authors present a report of a case of a tooth that was swallowed following extraction. Use of preventive techniques could have reduced the risk of this occurrence. CLINICAL IMPLICATIONS: Having a plan of action for such events is stressed. Preventive measures are reviewed. PMID- 12087836 TI - Oral pathology quiz #6. Calcified atheromas. PMID- 12087837 TI - Dentists talking negatively about dentists. PMID- 12087838 TI - Annual report of the president. PMID- 12087839 TI - Annual report of the Section on Public Health and Preventive Medicine. PMID- 12087841 TI - The vice-president's condition: a wake-up call to Congress. PMID- 12087840 TI - Final report of the college's presidential health task force. PMID- 12087842 TI - Annual report of the section on arts-medicine. PMID- 12087843 TI - Annual report on external affairs. PMID- 12087844 TI - Memoir: Lewis Lemon Coriell, M.D., Ph.D., 1911-2001. PMID- 12087845 TI - Annual report of the Katherine A. Shaw Division of Public Services. PMID- 12087846 TI - Annual report of the Mutter Museum and College Gallery. PMID- 12087847 TI - Annual report of the Historical Library. PMID- 12087849 TI - The College of Physicians of Philadelphia Financial Report FY2000. PMID- 12087848 TI - Annual report of the Francis C. Wood Institute for the History of Medicine. PMID- 12087850 TI - Annual report of the executive director FY2001. PMID- 12087851 TI - Carrying on when president cannot: exhibition raises questions about details in transfer of power. PMID- 12087852 TI - Opening remarks given at the college forum on presidential health. PMID- 12087853 TI - Deception and presidential disability: an historical analysis. PMID- 12087854 TI - Annual report of the Section on Medical History. PMID- 12087856 TI - Presidential disability and the 25th Amendment. PMID- 12087855 TI - Resuscitating the twenty-fifth amendment: mandating a second opinion for the determination of presidential disability. PMID- 12087857 TI - Health promotion, equity and access to information. PMID- 12087858 TI - Challenges and opportunities in strengthening the evidence base for mental health promotion. PMID- 12087859 TI - Assessing the health needs of young people. PMID- 12087860 TI - European training in health promotion: quality assurance based on collaboration and empowerment. PMID- 12087861 TI - Health promoting schools evidence for effectiveness workshops report. PMID- 12087862 TI - Management of nonsyndromic craniosynostosis. AB - In approximately 1 in 1000 live births in the United States, an infant has some variant of a facial, skeletal, or craniofacial deformity. If cleft lip and palate deformities are included, the incidence is greater. Timing of the surgical management of these patients has been advocated from the first few weeks after birth until well into the second decade. Many of these patients require multiple, staged procedures that involve movements of the bone and soft tissue from the intracranial and extracranial approaches. The surgical approach to most of these congenital deformities was radically changed by techniques introduced to the United States by Paul Tessier of France in 1967. From his imaginative intracranial and extracranial approaches, numerous advances have been made that facilitate the care of most of these children. More recently, additional advances in pediatric anesthesia and biodegradable plating systems have improved the management of these complex craniomaxillofacial deformities. PMID- 12087863 TI - Management of syndromic craniosynostosis. AB - Normalization of craniofacial malformation in syndromic craniosynostosis patients is an extended process. The care starts at birth and ends in adolescence. Multiple surgeries may be required for each deformity. The relationship among patient, family, and doctor team is important because they must understand the complete processes of management. Psychologists provide an important support mechanism. PMID- 12087864 TI - Monobloc and facial bipartition osteotomies. PMID- 12087865 TI - Frontoorbital reconstruction. AB - The care of the patient with craniosynostosis is perhaps the most challenging problem in craniofacial surgery. The syndromes vary in their presentation and severity; however, many of the structural and functional sequelae have common characteristics. The multifaceted medical problems of these children are particularly well suited to a multidisciplinary team approach to their care. The surgeons must be aware of the most pressing health concerns of these patients and incorporate those concerns into a well-formulated phased plan of comprehensive reconstruction. The overall goals of treatment of syndromic and nonsyndromic craniosynostosis continually should guide the care of these exceptional children. Those goals are to provide a predictable and durable reconstruction that restores facial form, reestablishes symmetry, attains the best results in the least number of operations, and infuses the patient with a sense of emotional and physical health. PMID- 12087866 TI - Endoscopic treatment of craniosynostosis. AB - We are entering a new era of craniosynostosis repair. When detected early, endoscopic skull remodeling, combined with a postoperative external skull-molding device, gives an excellent long-standing reconstruction of the cranial skeleton. This technique diminishes the morbidity of the operation and decreases the overall cost. It does not replace classic plate and screw cranial vault reconstruction in the older patient but is a useful weapon in the armamentarium of the craniofacial surgeon for the treatment of craniosynostosis in the neonatal period. PMID- 12087867 TI - Transfacial approaches to the cranial base. PMID- 12087868 TI - Dermoid cysts, gliomas, and encephaloceles: evaluation and treatment. AB - The fronto-orbitonasal malformations of encephaloceles, dermoid sinus cysts, and gliomas represent a continuum of neuroectodermal anomalies. The differentiation between them and other similar-appearing lesions is essential for effective management to proceed. Obtaining a reliable history, completing a careful physical examination, and obtaining accurate radiographic documentation represent the first steps. Establishing the timing, staging, and specific surgical techniques for management of a fronto-orbitonasal encephalocele remains as much an art as a science. PMID- 12087869 TI - XIX TNAI Biennial (68th) Conference, Goa, 2001: a report. PMID- 12087871 TI - World AIDS Day. PMID- 12087870 TI - Role of infection control nurse in the surveillance of nosocomial infections. PMID- 12087872 TI - Testosterone and estradiol modulate TNF-alpha-induced expression of adhesion molecules in endothelial cells. AB - Cytokine-activated endothelial expression of adhesion molecules plays an important role in immune responses. In the present study, we investigated the influences of testosterone and 17 beta-estradiol on tumor necrosis factor-alpha (TNF-alpha)-induced expression of adhesion molecules in human umbilical vein endothelial cells (HUVEC). HUVEC were incubated with TNF-alpha, testosterone or 17 beta-estradiol separately, or in a combination of TNF-alpha plus testosterone or 17 beta-estradiol. The expression of E-selectin, vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) was evaluated at 3, 6, 12 and 24 h following exposure by flow cytometric analysis. The results showed that although testosterone or 17 beta-estradiol did not affect the expression of these adhesion molecules in unstimulated HUVEC, both of them transiently increased the expression of E-selectin and VCAM-1 in TNF-alpha stimulated HUVEC. Neither testosterone nor 17 beta-estradiol affected the expression of ICAM-1 induced by TNF-alpha. It is concluded that both testosterone and 17 beta-estradiol increase TNF-alpha-induced expression of E-selectin and VCAM-1 in endothelial cells and these facts might indicate a mechanism by which gonadal hormones can indirectly enhance immune responses. PMID- 12087873 TI - The effect of losartan on calcium uptake by bovine aortic media in vitro. AB - The action of losartan, an AT1-angiotensin II receptor antagonist, on Ca2+ uptake by bovine aortic media in vitro was studied. This action was compared with that of nifedipine, a Ca2+ entry blocker. Ca2+ uptake values were obtained by measuring small slices of thoracic aorta tissue incubated in Kreb's solution in the presence of 45Ca. After preparation stimulation with angiotensin II (10 microM), losartan (1 microM and 10 microM) was shown to have a weaker inhibiting effect on Ca2+ uptake than nifedipine in equal concentrations. It is known that angiotensin II contracts smooth muscle by mobilizing intracellular Ca2+. The stimulating action on Ca2+ entry might be a secondary mechanism, which was studied for the first time in tissue slices derived from the aorta media. Results seem to be reasonable, since nifedipine also inhibited the spontaneous entrance of Ca2+, while losartan's action was focused only on the effects of angiotensin II. PMID- 12087874 TI - Pica in mice as a new model for the study of emesis. AB - In general, rats and mice have not been used in research on emesis because they do not vomit. However, emetogenic stimuli such as anticancer drugs, apomorphine, copper sulfate and rotation induced pica, a behavior characterized by eating nonfood substances such as kaolin, in rats. We also found that cisplatin induced pica in mice, but it was rather difficult to determine the exact kaolin consumption in this species. In this study, we prepared kaolin pellets mixed with carmine, a dye not absorbed in the gastrointestinal tract, and estimated kaolin consumption by determination of carmine excreted in feces. Cisplatin (5 mg/kg) caused a significant increase in kaolin consumption (saline: 0.15 +/- 0.08 g vs. cisplatin: 0.45 +/- 0.16 g) and pretreatment with the 5-HT3 receptor antagonist, ondansetron (2 mg/kg), suppressed the increased consumption (vehicle: 0.33 +/- 0.05 g vs. ondansetron: 0.13 +/- 0.04 g). These findings suggested that the exact kaolin consumption could be quantified by the determination of carmine in feces and that mice could be useful for studying emesis. PMID- 12087875 TI - Involvement of opioid delta (delta)- and kappa (kappa)-receptors in ischemic preconditioning in a rat model of myocardial infarction. AB - There is controversy in the literature regarding the involvement of opioid delta (DOP, OP1)- and kappa (KOP,OP2)-receptors in ischemic preconditioning (IPC). Previous studies on this subject in our laboratories and elsewhere have been performed on either isolated heart muscles of experimental animals, or in open heart surgery rats. To highlight this problem, we introduced an in vivo model of myocardial infarction in rats, which not only allowed electrocardiographic and enzymatic evaluation, but also morphometric assessment of myocardial infarction. In addition to these parameters, a direct receptor ligand study was undertaken, using [3H]-DPDPE, a specific opioid delta-receptor ligand. In our pharmacodynamic studies, we used the selective opioid delta-receptor agonist D-Ala2,D-Leu5 enkephalin (DADLE) and antagonist natrindole. For the evaluation of opioid kappa receptors, the selective opioid agonist U-50488H and antagonist nor-BNI were employed. Ischemic preconditioning showed the best beneficial effect, compared with pharmacological stimulation of either opioid delta- or kappa-receptors. In normal rat myocytes, two types of opioid delta-receptors exist, namely, low affinity and high-affinity opioid receptors. In acute myocardial infarction (30 min ischemia), the low-affinity type opioid receptors disappeared, most likely as a result of receptor downregulation due to an excessive release of enkephalins. There was no change in the density of the high-affinity opioid receptor type, but their affinity significantly increased (p < 0.05) by 58%. The radioligand receptor studies showed that opioid delta 1-receptor type was involved not only in triggering, but also in maintaining, the preconditioned state. On the basis of our pharmacodynamic studies, we suggest that both opioid delta 1- and kappa receptors are involved in the phenomenon of IPC, but with different effects. After 30 min of left coronary artery occlusion, opioid delta-receptor agonist DADLE decreased the infarct size/area at risk from 59.80% in control, untreated, infarcted rats, to 20.40% in treated rats, without a significant effect (p > 0.05) on the occurrence of early cardiac arrhythmias. Opioid kappa-receptor agonist U-50488H produced an opposite effect on the myocardium. It decreased the infarct size/area at risk by 44%, decreased occurrence of early arrhythmias by 77% and also decreased ventricular ectopic beats by 80%. The opioid delta- and kappa-receptor agonists used in this study significantly reduced (p < 0.05) early (2 h) postinfarction mortality by 22% and 19%, respectively. Further studies are in progress to differentiate between the role of opioid kappa 1- and kappa 2 receptors and the molecular mechanisms of the effects of both opioid delta- and kappa-receptors. PMID- 12087876 TI - Effect of melatonin and phenytoin on an intracortical ferric chloride model of posttraumatic seizures in rats. AB - The present study was carried out to investigate the effect of melatonin, a potent antioxidant, and phenytoin, a conventional antiepileptic, against FeCl3 induced posttraumatic seizures. Male Wistar rats weighing 200-250 g were implanted with epidural electrodes and allowed to recover. After the recording of baseline EEG, FeCl3 (5 ul, 100 mM) was administered intracortically over a period of 5 min and EEG was monitored for 2 h. Subsequently, rats were sacrificed to estimate oxidative stress, i.e., the amount of malondialdehyde (MDA) in whole brain tissue. A sham group was run parallel with saline (pH adjusted), and a similar protocol for EEG recording and estimation of oxidative stress was followed. FeCl3-treated animals exhibited epileptiform EEG changes (high amplitude sharp waves of increased frequency and polyspikes) within 15 min, which continued throughout the period of observation. MDA levels were found to be significantly elevated as compared to the sham group. Melatonin (50 mg/kg i.p.) administered 30 min before FeCl3 injection delayed the onset of appearance of epileptiform EEG changes, while at a 100 mg/kg dose of there was complete protection, as none of the animal exhibited epileptiform EEG discharge. Brain MDA levels were also significantly reduced in melatonin (50 and 100 mg/kg dose) treated animals as compared to the vehicle-treated FeCl3-injected rats. In the phenytoin group, all animals showed epileptiform EEG discharge. However, phenytoin at both 50 and 100 mg/kg dose delayed the onset of epileptiform EEG discharge. There were no significant changes in brain MDA levels in the phenytoin treated group as compared to controls. Melatonin and phenytoin at doses of 100 mg/kg did not show any sign of motor impairment as observed during the rota-rod test. These findings showed a superior protective effect of melatonin over phenytoin in an intracortical FeCl3 model of posttraumatic epilepsy. PMID- 12087877 TI - 1-(1-naphthyl)-piperazine, a mixed 5-HT1A and 5-HT2A/2C receptor ligand, elicits an anxiolytic-like effect in the open-field test without changes in 5-HT metabolism. AB - The effects of acute 1-(1-naphthyl)-piperazine (1-NP, 2 mg/kg i.p.) on rat open field and social interaction behaviors were studied followed by postmortem and in vivo microdialysis measurement of serotonin (5-HT) and 5-hydroxyidolacetic acid (5-HIAA) content. 1-NP treatment elicited an anxiolytic-like effect in the open field test, which was not modified by citalopram (5 mg/kg i.p.) challenge. A statistically insignificant tendency toward prolongation of the social interaction time was also found. The only significant change in the postmortem experiment was found in the striatum: 5-HIAA content was reduced after combined 1 NP plus citalopram treatment. Using the in vivo microdialysis technique, no difference in the 5-HT or 5-HIAA output between the treatment groups was found in the frontal cortex of anaesthetized rats. Our present study demonstrates the anxiolytic-like properties of 1-NP in the open-field test, but this effect is irrelevant to changes in 5-HT metabolism. PMID- 12087878 TI - Gateways to Clinical Trials. AB - Gateways to Clinical Trials is a guide to the most recent clinical trials in current literature and congresses. The data in the following tables has been retrieved from the Clinical Studies knowledge area of Prous Science Integrity, the world's first drug discovery and development portal, and provides information on study design, treatments, conclusions and references. This issue focuses on the following selection of drugs: Abiciximab, acetylcholine chloride, acetylcysteine, alefacept, alemtuzumab, alicaforsen, alteplase, aminopterin, amoxicillin sodium, amphotericin B, anastrozole, argatroban monohydrate, arsenic trioxide, aspirin, atazanavir, atorvastatin, augmerosen, azathioprine; Benzylpenicillin, BMS-284756, botulinum toxin type A, botulinum toxin type B, BQ 123, budesonide, BXT-51072; Calcium folinate, carbamazepine, carboplatin, carmustine, ceftriaxone sodium, cefuroxime axetil, chorionic gonadotropin (human), cimetidine, ciprofloxacin hydrochloride, cisplatin, citalopram hydrobromide, cladribine, clarithromycin, clavulanic acid, clofarabine, clopidogrel hydrogensulfate, clotrimazole, CNI-1493, colesevelam hydrochloride, cyclophosphamide, cytarabine; Dalteparin sodium, daptomycin, darbepoetin alfa, debrisoquine sulfate, dexrazoxane, diaziquone, didanosine, docetaxel, donezepil, doxorubicin hydrochloride liposome injection, DX-9065a; Eberconazole, ecogramostim, eletriptan, enoxaparin sodium, epoetin, epoprostenol sodium, erlizumab, ertapenem sodium, ezetimibe; Fampridine, fenofibrate, filgrastim, fluconazole, fludarabine phosphate, fluorouracil, 5-fluorouracil/epinephrine, fondaparinux sodium, formoterol fumarate; Gabapentin, gemcitabine, gemfibrozil, glatiramer; Heparin sodium, homoharringtonine; Ibuprofen, iloprost, imatinib mesilate, imiquimod, interferon alpha-2b, interferon alpha-2c, interferon-beta; KW-6002; Lamotrigine, lanoteplase, metoprolol tartrate, mitoxantrone hydrochloride; Naproxen sodium, naratriptan, Natalizumab, nelfinavir mesilate, nevirapine, nifedipine, NSC-683864; Oral heparin; Paclitaxel, peginterferon alfa 2b, phenytoin, pimecrolimus, piperacillin, pleconaril, pramipexole hydrochloride, prednisone, pregabalin, progesterone; Rasburicase, ravuconazole, reteplase, ribavirin, rituximab, rizatriptan, rosiglitazone maleate, rotigotine; Semaxanib, sildenafil citrate, simvastatin, stavudine, sumatriptan; Tacrolimus, tamoxifen citrate, tanomastat, tazobactam, telithromycin, tenecteplase, tolafentrine, tolterodine tartrate, triamcinolone acetonide, trimetazidine, troxacitabine; Valproic acid, vancomycin hydrochloride, vincristine, voriconazole, Warfarin sodium; Ximelagatran, Zidovudine, zolmitriptan. PMID- 12087879 TI - [Effectiveness of ulcer treatment with electromagnetic radiation of extremely high frequency (EHF therapy) and some mechanism of its therapeutic action]. AB - A clinical response to and some mechanisms of therapeutic action of extremely high frequency (EHF) therapy were studied in 132 patients with exacerbation of duodenal ulcer vs routine pharmacological treatment. EHF-therapy was used alone and in combination with famotidin (antisecretory drug) and norfloxacine (antibacterial drug). EHF monotherapy proved highly effective in duodenal ulcer exacerbation. It normalizes secretory and motor functions of the stomach, suppresses initially high activity of free radical lipid oxidation, corrects abnormal vegetative and psychoemotional status of the patients, moderately potentiates the antihelicobacter effect of antibacterial drugs. These effects are produced due to specific action of EHF therapy: mobilisation of sanogenesis mechanisms, correction of mechanisms of adaptive regulation and self regulation at different levels. Additional administration of antisecretory and antibacterial drugs improved immediate but deteriorates long-term response to EHF-therapy. PMID- 12087880 TI - [Oxygen transport and immune status in patients with congenital valvular heart disease]. AB - Oxygen transport, its metabolic maintenance and immune status were studied in 17 patients with congenital valvular heart disease (CVD) having compensated (n = 8, group 1) and decompensated (n = 9, group 2) defects of hemodynamics. CVD patients with decompensated central hemodynamics and progressing hypoxia had impaired compensatory rearrangement of oxygen transport system. Accumulation of intracellular lactate, low activity of basic energetic cycles of blood cells most evident in decompensated CVD was observed in both the groups. In conditions of severe energy-structural deficiency and impaired function of oxygen transport systems, CVD patients develop secondary immune deficiency presenting with depression of basic immunoregulatory subpopulations of T- and B- cellular immunity (CD2, CD3, CD4, CD5, CD8, CD16, CD22). PMID- 12087881 TI - [The response to normobaric hypoxia and its duration in arterial hypertension patients as shown by a prospective study]. AB - A placebo-controlled parallel trial studied an antihypertensive action of interrupted normobaric hypoxia (INH) in 63 male patients with arterial hypertension (AH) of the first degree and low and moderate risk of vascular complications. The placebo group consisted of 18 patients with AH of the first degree matched by age, the disease duration, initial level of systolic and diastolic pressure. An INH course consisted of 15 daily procedures. INH monotherapy implied oxygen content lowering to 10%. A 6-month follow-up was made with the use of 24-h AP monitoring. The INH course resulted in a significant decrease in systolic and diastolic AP, total peripheral vascular resistance. A morning rise of systolic pressure became slower. A favourable effect of normobaric hypoxia on AP 24-h rhythm was observed. A good hypotensive effect was achieved in 85.7% patients. A 6-month follow-up confirmed a long-term hypotensive effect of INH. PMID- 12087882 TI - [Analysis of lipid plasma spectrum and basic parameters of red cell membranes in patients with metabolic syndrome and ischemic heart disease]. AB - Specific features of lipid plasma spectrum and principal parameters of red cell membranes are characterized in patients with metabolic syndrome (MS) and chronic stable ischemic heart disease (IHD). 109 patients with metabolic syndrome (diabetes mellitus type 2, arterial hypertension, abdominal obesity and dyslipidemia) were divided into 2 groups: with and without IHD. MS patients with IHD had marked defects of lipid metabolism with hypercholesterolemia, high levels of triglycerides, LDLP cholesterol, low level of HDLP cholesterol. Lipid plasma spectrum in MS patients with IHD vs those without coronary atherosclerosis was characterized by a significantly lower level of apo A1. In red cell membranes these patients had lower fractions of esterified cholesterol combined with high intensity of lipid peroxidation. PMID- 12087884 TI - [Rheumatic manifestations of Takayasu's disease]. AB - Rheumatic manifestations of nonspecific aortic arteritis are described with a special focus on analysis of early symptoms of the disease. PMID- 12087883 TI - [Optimization of cardiovascular function by peptide bio-regulators]. AB - Effects of peptide bioregulators (thymaline, epithalamine, cortexine) on systemic hemodynamics in cardiovascular and cerebrovascular affections were studied. Assessment of functional stability and quality of cardiovascular system regulation was made according to methodological principles of automatic regulation theory. Peptide bioregulators showed high effectiveness in the above patients who markedly improved their hemodynamic parameters. PMID- 12087885 TI - [Changes in clinical and laboratory findings on oxidation metabolism in lymphocyte membranes of rheumatoid arthritis patients on synchronous programmed intensive therapy]. AB - The study of the effects of intensive treatment on activity of free-radical oxidation in lymphocyte membranes was conducted in 15 rheumatoid arthritis (RA) patients with systemic involvement, stage II-III activity who had failed previous basic therapy because of unefficiency or intolerance. The synchroneous programmed intensive therapy was performed in 3 stages. Stage 1: three sessions of plasmapheresis with 3 day intervals, synchronous introduction of 30 mg methotrexate and 500 mg methylprednisolone. Stage 2: the same procedures once a week for the following 3 weeks. Stage 3: a monthly intensive therapy for 3 months. After the end of stage 3 all the patients received methotrexate 7.5 mg weekly per os for 6 months. The above intensive therapy of RA patients had an antiinflammatory effect, changed peroxidation reactions in lymphocyte membranes by lowering the level of pro-oxidants and stimulation of antioxidant system. No positive changes in the lipid status occurred. PMID- 12087886 TI - [Thrombophilia, venous thromboses and their treatment]. AB - Thromboses are leading among causes of death worldwide. Arterial thromboses--the underlying cause of myocardial infarctions, strokes and acute ischemia--develop due to a defect of atherosclerotic plaque. Venous thromboses and associated venous thromboembolism killing one of 1000 world population develop in unaffected vascular wall. Recent data show that 70% cases of venous thromboembolism can be prevented by identification of thrombophilia markers--inheritable conditions associated with thromb formation. Current information on variants and prevalence of thrombophilia is given as well as recommendations on venous thromboembolism prevention. PMID- 12087887 TI - [The response to combined therapy of hepatic encephalopathy in patients with hepatic cirrhosis]. AB - Combined therapy with gepa-merz (L-ornitine-L-aspartate) and lactulose (a synthetic disaccharide) was given to 40 patients with hepatic encephalopathy and toxic hepatic cirrhosis in decompensation. Combination of gepa-merz with lactulose was effective in the above patients. Moreover, it is an optimal one as it has a double impact on ammonia toxicity which is a basic neurotoxin in pathogenesis of hepatic encephalopathy. PMID- 12087888 TI - [Clinico-laboratory and morphological parallels in hepatic echinococcosis]. AB - The trial included 50 patients with hydatid echinococcosis of the liver and 5 control patients operated urgently for extrahepatic pathology. The patients underwent clinical and biochemical tests, ultrasound investigation of the liver. Structural and functional changes in the liver were analysed with reference to the stage of the echinococcal cyst. Morphological picture of the liver was studied by light microscopy, its functional picture--by activity of AsAT and AlAT in hepatic tissue. Morphological and biochemical tests were made on hepatic biopsies obtained from the pericystic and far from the cyst zones. The most severe histological changes affecting all structures of the liver were registered in the parasite's life cycles II and III. AsAT and AlAT changes in the liver were not related to the biological cycle of the parasite; echinococcal patients' liver contained them in lower concentrations than that of control subjects. Moreover, blood serum levels of AsAT and AlAT are higher than in liver tissue. Thus, growth of the echinococcal cyst in the liver is accompanied with marked morphofunctional disorders. This necessitates early detection and surgical treatment of hepatic echinococcosis. To prevent postoperative complications, adequate preoperative preparation and postoperative intensive treatment must be conducted. PMID- 12087889 TI - [Enhancement of antioxidant status of the elderly patients with ischemic heart disease in response to amino acid composition of MP-33 in combined therapy with trimetazidine]. AB - Effects of amino acid composition MP-33 in combination with trimetazidine were studied in metabolic therapy of elderly patients (age 63.3 +/- 1.7 years) with ischemic heart disease (angina pectoris functional class II-III). MP-33 (100 mg 3 times a day subling.), trimetazidine (20 mg 3 times a day per os), trimetazidine + amino acid composition MP-33 in the above doses in addition to basic therapy were given to 30, 15 and 15 patients, respectively. Combined therapy with trimetazidine and MP-33 significantly activated antioxidant enzymes and depressed lipid peroxidation due to GSH-dependent regulation of cellular redox status by MP 33. PMID- 12087890 TI - [Long-term dirotone treatment of patients with chronic obstructive bronchitis complicated by cor pulmonale]. AB - 52 patients with chronic obstructive bronchitis complicated by chronic decompensated cor pulmonale participated in the trial of dirotone. Effects on the clinical course and hemodynamics were assessed in the course and long-term treatment. A dirotone course was found to benefit central and pulmonary hemodynamics while a long-term dirotone treatment relieves symptoms of cardiac failure and raises life quality in the absence of negative effects on blood biochemistry, electrolytic balance and external respiration function. PMID- 12087891 TI - [Aortic dissecting aneurysms: clinical masks, differential diagnosis]. AB - 101 necropsies of aortic dissection (AD) made in 1991-2000 have been analysed. Among the deceased men prevailed (76.3%), 23(22.8%) of them were under 50 years of age. An accurate intravital diagnosis was made only in 40.6 patients. The most common clinical masks were the following: coronary, valvular, vascular, gastrointestinal, pulmonary, thromboembolic, mediastinal, cerebral, renal and anemic. Most often AD masks as acute myocardial infarction. The knowledge of encountered clinical masks of AD promoted improvement of early AD diagnosis to correct AD surgically and reduce lethality. PMID- 12087892 TI - [Introduction of tele-medical electrocardiographic system in regional general hospital]. AB - The paper reviews the experience in setting up and functioning of telemedical electrocardiographic system in a regional hospital with utilization of current potential of telephone ECG transmission-reception: individual ECG registers transmitters including devices with "a reverse loop". It is shown that ECG registration by patients themselves with its transmission by telephone facilitates diagnosis of rare paroxysmal conditions and follow-up of cardiological outpatients. Changes in the number of ECGs made for 25 years in the regional center of remote cardiological diagnosis is analysed. PMID- 12087893 TI - [Two cases of diagnosis of right ventricular myocardial infarction]. AB - Right ventricular myocardial infarction (RVMI) occurs rather rarely, under the mask of various diseases and is not easy to detect. Hypo-vate prognosis. Intravital diagnosis of RVMI was successful in 4 patients. Location of the lesion and its extent were determined. This article reports two cases of the four. The disease onset manifested with dyspnea, dry cough, attacks of night asphyxia then symptoms of right ventricular failure arose and intensified. The clinical picture, typical ECG signs suggested the RVMI diagnosis. However, its verification, precise localization and estimation of the lesion size were possible only after balanced radioventriculography. PMID- 12087895 TI - [M. Ia. Mudrov: an outstanding figure in Russian medicine. 225 years since birth]. PMID- 12087896 TI - [Statins in clinical practice]. AB - The review covers principles of treatment with statins, their pharmacokinetics, characterizes six most usable in clinical practice statins and one novel drug- rozuvastatin from a "superstatins" group. Statins proved effective in primary and secondary prophylaxis of coronary atherosclerosis, secondary prevention of ischemic stroke and diseases of peripheral arteries with intermittent claudication, prevention of Alzheimer's disease. Statins are indicated in dyslipidemia in diabetes mellitus type 2, nephrotic syndrome and renal insufficiency. Further studies are needed on statins' effects on hypercholesterolemia, their antiinflammatory and immunomodulating properties, the ability to enhance revascularization of ischemic tissue and stimulate proliferation of osteoblasts in osteoporosis. PMID- 12087897 TI - [Prevalence and risk factors in osteoarthrosis in a rural population of the Sakha Republic (Iakutiia)]. AB - AIM: To study epidemiology of osteoarthrosis among native population living in rural North of Russia. MATERIAL AND METHODS: Population of four villages of the Vilyui region of Sakha Republic (1216 citizens) was covered with a one-stage survey. Population of three villages were Yakuts, one village--evenks. RESULTS: Prevalence of osteoarthrosis among native population of rural Yakutia is 18.0% (definite-9.4%, suspected--8.6%). General and specific osteoarthrosis risk factors linked with climatic-geographic local peculiarities were determined. CONCLUSION: Prevalence of osteoarthrosis among rural citizens of Yakutia is higher than among urban population and depends more on micro- and macroclimatic conditions and occupational hazards. PMID- 12087898 TI - [Prevalence of joint hypermobility among an adult population in Moscow]. AB - AIM: To elucidate joint hypermotility (JHM) among adult population of Moscow by age and sex. MATERIAL AND METHODS: Health survey covered 572 examinees (266 females and 306 males) in four age groups (16-20, 21-30, 31-40 and 41-50 years). JHM was assessed by Baton's scale. RESULTS: Prevalence of JHM in Moscow was similar to that in Europe. JHM occurred more frequently at young age and in women. The score 4-6 by Baton in Russian population (50% of 16-30-year-old women and 58% of 16-20-year-old males) is a variant of normal standard. PMID- 12087900 TI - [Soluble adhesion molecules in antiphospholipid syndrome associated with systemic lupus erythematosus and in primary antiphospholipid syndrome]. AB - AIM: To evaluate clinical implications of measurements of the level of soluble cell molecules of adhesion (sCMA) in systemic lupus erythematosus (SLE) with antiphospholipid syndrome (APS) and primary APS (PAPS). MATERIAL AND METHODS: Serum levels of sP-selectine, sE-selectine, sVCAM-1 were determined with enzyme immunoassay (R&D, USA) in 23 SLE with APS, 15 SLE patients free of APS and 19 patients with PAPS. RESULTS: Mean levels of sE-selectine and sVCAM-1 in SLE patients with APS, PAPS and SLE was higher than in donors. Elevated mean levels of sP-selectine were observed only in SLE patients free of APS. These patients also showed a correlation between sVCAM-1 level and the disease activity (p < 0.01). CONCLUSION: The development of immunopathological process in APS is associated with increased concentration of sCMA. PMID- 12087899 TI - [Environmental effects on the course of active rheumatism in a migrant Siberian population]. AB - AIM: To characterize antistreptococcal immune response in healthy subjects with cryoglobulinemia and rheumatic manifestations as well as a rheumatism course in population who came to Siberia to build and operate Baikal-Amur railway. MATERIAL AND METHODS: Siberian migrants were examined. Among them there were 203 patients with rheumatic fever and rheumatic heart disease, and 1275 healthy subjects. 65 patients had active rheumatism, 106 healthy subjects had rheumatic manifestations and cryoglobulinemia with adaptation cryoglobulinemia syndrome. All the patients underwent examination of the cardiovascular system, standard laboratory tests and tests for antistreptolysine-0, antistreptokinase, antistreptohyaluronidase, streptococcal antigen, microbiological tests. The diagnosis of rheumatic fever was established on the basis of Kisel-Jones-Nesterov and revised Jones criteria. RESULTS: The adaptation cryoglobulinemia syndrome in healthy persons was associated with cardialgia, arthralgia, lumbalgia, myalgia, lymphadenopathy and Raynaud's phenomenon. Patients with rheumatic fever and adaptation cryoglobulinemia syndrome were predisposed to chronicity of primary rheumatic carditis, development of compound and combined valvular defects, arrhythmia and hemodynamic disturbances. CONCLUSION: The presence of the adaptation cryoglobulinemia syndrome aggravates the course of rheumatic fever and promotes its chronicity. PMID- 12087901 TI - [Mutation of a 5,10-methylenetetrahydrofolate reductase gene in systemic lupus erythematosis and antiphospholipid syndrome]. AB - AIM: To study prevalence of mutation C677T in gene 5.10-MTHFR in systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APS) as well as in persons free of symptoms of systemic diseases of the connective tissue. MATERIAL AND METHODS: 85 patients participating in the study were divided into three groups: those with SLE (n = 17), with SLE + APS (n = 42), with primary APS (n = 26). The control group consisted of 30 persons without SLE or APS. 55% of the examinees had thrombotic complications of different location. The diagnosis of the mutation was made using DNA isolated from the peripheral blood with standard methods and polymerase chain reaction. Allele (homozygous or heterozygous) condition of the mutation was confirmed by means of allele-specific primers. RESULTS: Mutation C677T in MTHFR gene was found in 40 of 85 patients (47%); 11(27.5%) had a homozygous variant, 29(72.5%)--heterozygous. C677T mutation occurred in 5 of 17 SLE patients (29%), it was in all the cases heterozygous. In primary and secondary APS mutation was detected in 51.5% (35 of 68 patients). Recurrent thrombosis occurred more frequently in patients with mutation MTHFR. Three and more episodes of thrombosis were registered in 17 of 40 patients with mutation C677T against 9 of 44 patients without the mutations (p = 0.04). CONCLUSION: Relationship between elevated blood levels of APL and MTHFR mutation points to the fact that this genetic marker is an additional thrombogenic factor in APS. Mutation C677T in MTHFR gene in APS patients correlates with recurrent thrombosis. PMID- 12087902 TI - [Clinical and pathogenic significance of tumor necrosis factor-alpha in systemic lupus erythematosus]. AB - AIM: To examine the level of tumor necrosis factor-alpha (TNFa) in the sera from patients with systemic lupus erythematosus (SLE) and its clinical and pathogenic role. MATERIAL AND METHODS: TNFa was measured with ELISA (Immunogenetics N.V., Belgium) in the sera from 147 SLE patients (70 men and 77 women). The results were compared with those of 20 healthy subjects. RESULTS: TNFa elevated concentrations were found in 49% examinees with SLE. Concentration of TNFa, SLE activity and development of antiphospholipid syndrome correlated. CONCLUSION: The findings indicate that TNFa is involved in pathogenesis of SLE. Quantitation of TNFa may serve a useful tool for monitoring SLE activity. PMID- 12087903 TI - [Chromosomal aberrations in lymphocytes of patients with systemic lupus erythematosus during cytostatic therapy]. AB - AIM: To study cytogenetics of peripheral blood lymphocytes in patients with systemic lupus erythematosus (SLE) in the course of therapy with a selective immunodepressant sandimmun and universal cytostatic cyclophosphamide. MATERIAL AND METHODS: 30 women with lupus-nephritis and nephrotic syndrome received sandimmun in a dose 2.5-7 mg/kg plus 10-20 mg prednisolone for a month with gradual lowering of sandimmun dose to 2.5-5 mg/kg and prednisolone to 0-10 mg/day. 10 patients with lupus-nephritis were given cyclophosphamide in a dose 1000 mg as a single intravenous drip once a month for 6 months in combination with 30-40 mg prednisolone for a month with the dose reduction to 20 mg/day. Sandimmun and cyclophosphamide effects on mutagenesis were studied in 72-120-h PHA-stimulated lymphocyte cultures. Chromosomal aberrations (CA) were assessed according to A.F. Zakharov's classification and proposals of International workshop in Melburn (1993). RESULTS: Sandimmun therapy on day 14 did not increase CA frequency but led to appearance of "premature chromosome condensation phenomenon" (PCC). PCC diminished after one month of sandimmun therapy and disappeared after 3-6 months. Cyclophosphamide provoked a rise in the incidence of both chromatide aberrations and CA. CONCLUSION: Sandimmun does not increase CA incidence in lymphocytes. On the contrary, it promoted the fall in this incidence while syclophosphamide stimulated CA and chromatide aberrations. This trend was observed till the end of cyclophosphamide administration. PMID- 12087904 TI - [Antineutrophil cytoplasmic antibodies in patients with diffuse diseases of connective tissue]. AB - AIM: To investigate the significance of antibodies against neutrophil cytoplasmic antigens (ANCA) in patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) and systemic sclerosis (SS), their relations with the syndromes and laboratory indices. MATERIAL AND METHODS: The sera from 277 patients suffering from connective tissue disease and 31 healthy persons were examined. Of patients, 103, 126 and 48 had RA, SLE and SS, respectively. The patients in each group were subdivided into ANCA positive (ANCA+) and ANCA negative (ANCA-). The patients were matched within the groups by age, sex and disease duration. There were 41 such pairs in RA group, 23 in SLE and 13 in SS group. Questionnaires and laboratory tests (ANA, RF, a-DNA, a-MPO, a-Scl-70, a PR3, a-CL) were used in the examination. RESULTS: The sensitivity of ANCA in SLE patients group was as high as 58.7%, specificity--93.5%. In other groups ANCA were less frequent. ANCA were significantly associated with skin vasculitis and ANA prevalence but the disease activity in SLE was not related to this feature. Anemia and antibodies against cardiolipin were found significantly more frequently in ANCA positive RA group. In SS group the inverted clinical association with kidney damage was seen but a-DNA were more prominent in ANCA+ group. Subspecificity for a-MPO and a-PR3 and lactoferin by ELISA were revealed less often than a-ANCA by immunofluorescence. Only two SLE patients with a lactoferin antibodies were evidently different in prognosis while the other ones did not differ in the disease course within their group. CONCLUSION: The ANCA pattern in connective tissue diseases is found rather often but only few clinical and laboratory associations could be established such as skin vasculitis and ANA domination in SLE group, anemia and a-CL in RA group and a-DNA in SS ANCA+ groups. The validity of ANCA test is not so significant as it is in vasculitis patients. PMID- 12087905 TI - [Alterations of rheological properties of blood in systemic sclerodermia]. AB - AIM: To study hemorheological status in systemic sclerosis (SS) patients. MATERIAL AND METHODS: Macro- and microrheology of blood were investigated in 70 SS patients. The rate of spontaneous aggregation of red blood cells [T1(s)], durability of the largest units [Ia2.5(%)], general hydrodynamic durability of units [beta(s-1)], limit of fluidity [tau 0 (mPa)], Casson viscosity K (mPa/s) were determined. RESULTS: The total severity of hemorheological disturbances was 1.90 +/- 0.07 with T1 = 4.98 +/- 1.52s, beta = 62.76 +/- 19.32s, Ia 2.5 = -0.78 +/- 12.83%, tau 0 = 7.00 +/- 0.45 mPa. The Casson viscosity did not differ from normal levels. Plasma viscosity was 1.71 +/- 0.18 rel units, viscosity of sera 1.55 +/- 0.14 rel.units. The severity of hemorheological disturbances did not differ in cute and chronic courses but the parameter beta (72.0 +/- 4.5) was significantly higher (p < 0.05) in the acute course. The severity of hemorheological disturbances was highest in SS patients with cardiac disease (p < 0.05) and in 10 year SS duration. CONCLUSION: The course of SS is complicated by progressive hyperaggregation syndrome promoting development of visceral pathology. Active correction of hemorheology is pathogenetically validated in SS. PMID- 12087906 TI - [Hypocomplementary urticarial vasculitis with the mitral valve affection and complement-fixed antibodies to the myocardial antigens]. PMID- 12087907 TI - [Incidence rate of rheumatic diseases in Russian population: 10-year analysis]. AB - AIM: To analyze trends in rheumatic diseases (RD) incidence rate in population of Russia for a recent decade. MATERIAL AND METHODS: Medical RD statistics for all regions of Russia for 10 years (1990-1999) have been analysed. RESULTS: Annual number of primary patients with RD has increased for 1990-1999 from 9,147,000 to 12,378,000 (by 35.3%). The percentage of patients with acute rheumatic fever and chronic RD of the heart diminished from 6.5 to 2.9%. Most of the diseases constitute the diseases of osteomuscular system and connective tissue (DOMS). The number of new cases with rheumatic heart diseases (RHD) in adolescents and adults has risen by 50 and 40%, respectively. RHD are especially frequent in the Volga Vyatka and North-Caucasian regions (2.5 and 2.3 per 1000 against mean Russian 1.7). A total DOMS morbidity has increased for the recent decade by 41%. It is higher in the Central and North regions, in the West Siberia and lower in the North Caucasus, Far East, East Siberia. For the 7 years the above morbidity among children rose 2-fold, among adolescents--2.4-fold. Incidence of rheumatoid arthritis and ankylosing spondylarthritis in adult population of Russia has changed insignificantly while osteoarthrosis total and primary occurrence for 6 years increased by 78 and 58%, respectively. CONCLUSION: RD become a real challenge for health service of Russia. Changes in RD structure require correction of the rheumatic service activities. PMID- 12087908 TI - [Synchronous programmed intensive therapy of patients with severe rheumatoid arthritis]. AB - AIM: To evaluate efficiency and safety of intensive treatment program (synchroneous plasmapheresis, large-dose methotrexate and methypred) for patients with severe rheumatoid arthritis (RA). MATERIAL AND METHODS: 45 patients with highly active and progressive RA, systemic symptoms, corticosteroid dependence who had intolerance to standard therapy or had not responded to it were divided into 2 comparable groups. 25 patients of group 1 for a month got 6 plasmapheresis procedures with synchroneous intravenous injection of 40 mg of methotrexate and 250 mg of methypred. 20 patients of group 2 received pulse therapy with methypred (3 g) and methotrexate (200 mg). The intensive therapy was followed in all the patients with methotrexate in a dose 10-20 mg/week. RESULTS: One, six and twelve months after treatment patients of group 1 demonstrated a decrease in RA clinical activity and inflammation. In a year remission by ACR criteria was achieved in one-third of the patients. CONCLUSION: The sychroneous program of intensive therapy is highly effective in RA patients with vasculitis, ineffective standard therapy and corticosteroid dependence. PMID- 12087909 TI - [Current approaches to assessment of rheumatoid arthritis activity]. AB - AIM: To compare domestic criteria of RA activity and the disease activity scores (DAS). MATERIAL AND METHODS: Russian criteria of RA activity and DAS were used in 99 RA patients. RESULTS: It is shown that RA activity by DAS is higher than when it is assessed by criteria practiced in Russia. CONCLUSION: Further studies are necessary to examine clinical value of DAS criteria. PMID- 12087910 TI - [The role of suprasegmental and segmental autonomic disorders in the development of muscular syndrome of rheumatoid arthritis]. AB - AID: To investigate the role of suprasegmental (anxious disorders) and segmental vegetative disorders (dysfunction of neuromuscular transmission and cutaneous microcirculation) in pathogenesis of muscular syndrome of rheumatoid arthritis (RA). MATERIAL AND METHODS: MMPI, Beck's questionnaire, Spilberger's test were used in experimental-psychological assessment of RA patients' personality profile. A total of 60 healthy subjects and 340 RA patients were examined. Segmental vegetative disorders were studied by excitability of hand muscles (Prognoz-6 device) and cutaneous microcirculation (doppler flowmetry). RESULTS: RA patients showed suprasegmental disorders manifesting as frequent anxious disturbances (95%). Dysfunction of the segmental part of the autonomic nervous system was represented by various changes in excitability of hand muscles in response to weak currents, by abnormal regulation of cutaneous microcirculation. CONCLUSION: Neuromuscular and microcirculation disturbances participate in development of RA muscular syndrome. This is confirmed by a positive correlation coefficient. There was also a negative correlation between suprasegmental and segmental disorders. PMID- 12087911 TI - [Clinical efficacy of xefocam and its effect on arterial pressure and heart rhythm variability in patients with rheumatoid arthritis in combination with arterial hypertension]. AB - AIM: To try clinical response to xefocam, its safety, effects on arterial pressure and heart rhythm variability in rheumatoid arthritis (RA) patients with arterial hypertension (HT). MATERIAL AND METHODS: Xefocam (lornoxicam), a new non steroid antiinflammatory drug, was given for 12 weeks in a daily dose 12 mg/day to 44 RA patients (mean age 54.5 +/- 7.3 years). 24-h arterial pressure monitoring was made with Cardiotens-01 device. RESULTS: Xefocam in a dose 12 mg/day has shown good tolerance, a high analgetic and antiinflammatory effect as indicated by a positive response of articular syndrome, a significant fall of systolic arterial pressure, decreased heart rate, better heart rhythm variability. CONCLUSION: In hypertensive RA patients xefocam in a dose 12 mg/day proved effective and safe. PMID- 12087912 TI - [Levels of lipid peroxidation products and activity of antiradical defense enzymes in blood of patients with Bechterew disease]. AB - AIM: To study lipid peroxidation (LPO) in Bechterev's disease (BD) treated with antiinflammatory and antioxidant drugs. MATERIAL AND METHODS: LPO was estimated in 75 BD patients before medication and during this treatment. 30 healthy volunteers served control. RESULTS: Levels of LPO products was high but activity of antioxidant defense enzymes low in BD patients. CONCLUSION: Antiinflammatory drugs meloxicam, indometacin and an antioxidant drug triovit depressed LPO activity, improve the patients' condition and results of functional tests. PMID- 12087913 TI - [Approaches to late prognosis in patients with subacute infectious endocarditis]. AB - AIM: To ascertain late prognosis of subacute infectious endocarditis (IE), factors involved in the prognosis, formulate approaches to assessment of late prognosis. MATERIAL AND METHODS: A retrospective trial included 98 patients discharged from a hospital with diagnosis "subacute IE", treated without surgery or without endocarditis of the prosthesis. The diagnosis was verified at echocardiography. A mean follow-up 4.8 +/- 3.7 years. Clinical and echo-CG follow up findings were computer-processed. RESULTS: 5-year lethality was 31%. The patients died of cardiac failure (CF) and acute failure of cerebral circulation (84 and 16%, respectively). Late complications in the survivors were the following: severe CF (18 patients, 32%), moderate CF (18 patients, 32%), recurrent IE (12 patients, 20%). Such factors as IE variant (primary or secondary), affection of the aortic valve, severity of regurgitation, size of the left ventricle, CF progression in the acute phase proved to have a significant influence on late prognosis. CONCLUSION: The prevalent cause of death in IE is CF. The progression of CF depends on combination of the above factors. High risk group patients should be examined by a cardio-surgeon even in the absence of clinical symptoms of CF. PMID- 12087914 TI - [Plasma replacement in the treatment of ischemic heart disease]. AB - AIM: To investigate whether plasma replacement (PR) can raise efficiency of drug therapy of ischemic heart disease (IHD). MATERIAL AND METHODS: A 30-60% replacement of circulating plasma for salt or dextran using PF-05 unit has been performed in 324 patients 35-79 years of age with recurrent myocardial infarction and angina pectoris. A total of 520 PR procedures were performed. Biochemical, acid-base, coagulative, viscosity, microcirculatory blood parameters were taken, ECG and stress tests were made. RESULTS: PR resulted in a significant reduction in packed cell volume, total protein, fibrinogen, low density lipoproteins, total cholesterol and led to diminution of blood viscosity, acceleration of capillary blood flow, improvement of O2/CO2. Lowering of fibrinogen levels, number of platelets and their aggregation, enhancement of fibrinolytic blood activity created conditions for moderate controlled hypocoagulation. As shown by stress tests, two weeks after PR 60% of anginal patients of functional class IV can be transferred to class III. CONCLUSION: Because PR is beneficial by many parameters, it lessens the requirement in pharmacological support of patients with complicated IHD. PMID- 12087915 TI - [Two cases of successfully treated multifocal fibrosis]. PMID- 12087916 TI - [Vasomotor (nitritoid) reactions induced by sodium aurothiomalate and enap]. PMID- 12087917 TI - [Immunologic markers of atherosclerosis]. PMID- 12087918 TI - [Efficacy and perspectives of use of vasaprostan (prostaglandin E1) in rheumatology]. PMID- 12087919 TI - Influence of inhibition of adenosine uptake on the gamma-aminobutyric acid level of the ischemic rat brain. AB - Loss of inhibitory GABAergic modulation may induce hyperexcitability of neuronal elements and potentiate their vulnerability to excitotoxic injury. It has been also found that the adenosine system interacts with the GABAergic system during ischemia and anoxia where the adenosine receptors as well as the GABAergic receptors may conscript the same intracellular pathway to increase tolerance to ischemia. Therefore, it was aimed to study whether dipyridamole (CAS 58-32-2), an adenosine uptake inhibitor, or adenosine (CAS 58-61-7) could affect the rat brain gamma-aminobutyric acid (GABA, CAS 56-12-2) level after induction of cerebral ischemia, and to test their effect on the lactate dehydrogenase (LDH) activity of the ischemic rat brain. Ischemia was induced by bilateral clamping of the common carotid arteries for 60 min followed by reperfusion for other 60 min. Dipyridamole was administered alone and in combination with adenosine and the effect of the drugs on the brain GABA level as well as on LDH activity was determined. The results show that dipyridamole could protect the brain of rats against the ischemic insult, while adenosine when administered separately failed to influence the selected parameters. Protection of the rat brain by dipyridamole might be related to the elevated level of GABA. PMID- 12087920 TI - Inhibitory effects of alprostadil (prostaglandin E1) incorporated in lipid microspheres of soybean oil on intimal hyperplasia following balloon injury in rabbits. AB - The present study was performed to assess the inhibitory effects of alprostadil (CAS 745-65-3, prostaglandin E1, PGE1) incorporated in lipid microspheres (here in-after referred to as lipo PGE1; Palux inj.) on intimal thickening following balloon injury in the carotid artery of normal rabbits. Lipo PGE1 was given intravenously to animals twice a day at doses of 20 or 40 micrograms/kg/day from ballooning (day 1) until day 3, and at half these doses from day 4 to day 20. The carotid artery was removed for histopathological staining on the next day (day 21) after the last administration. Lipo PGE1 significantly reduced both the intimal/medial are (I/M) ratio and stenosis ratio by about half in the 40 micrograms/kg/day on day 21 after ballooning, compared with the vehicle group. Infiltration of macrophage, expression of proliferating cell nuclear antigen (PCNA)-positive cells was inhibited by the administration of lipo PGE1 on day 3 after ballooning. Adhesion of platelets to injured arterial walls was also inhibited on day 3. Lipo PGE1 at 40 micrograms/kg/day exerted more potent inhibitory effects on I/M and stenosis ratios and histopathological changes such as infiltration of macrophage and expression of PCNA-positive cells than at 20 micrograms/kg/day. These findings suggest that lipo PGE1 inhibits the intimal hyperplasia after balloon injury in rabbit carotid artery, possibly by inhibiting platelet functions. PMID- 12087921 TI - The effects of calcium channel antagonists on coronary nitrite outflow in isolated rat heart. AB - The aim of the study was to compare the effects of Ca2+ channel antagonists on coronary endothelial L-arginine/NO system in isolated rat heart. The hearts of male Wistar albino rats (n = 36, age 8 weeks, body mass 180-200 g) were perfused according to Langendorff technique at gradually increased coronary perfusion pressure (CPP) which induced flow-dependent NO release (nitrite outflow). The experiments were performed during control condition or in the presence of different Ca2+ channel antagonists: nifedipine (CAS 21829-25-4, 30 nmol/l), diltiazem (CAS 42399-41-7, 3 mumol/l), verapamil (CAS 52-53-9, 0.4 mumol/l) or amlodipine (CAS 88150-42-9, 100 nmol/l) were administered separately. Also, nifedipine or amlodipine were administered in combination with an inhibitor of nitric oxide synthase (NOS), L-NAME (NG-nitro-L-arginine-methylester, 30 mumol/l). Coronary flow (CF) varied in autoregulatory range from 3.93 +/- 0.25 ml/min/g wt at 50 cmH2O to 4.49 +/- 0.31 ml/min/g wt at 90 cmH2O. In autoregulatory range nitrite outflow varied from 1.80 +/- 0.22 nmol/min/g wt at 50 cmH2O to 2.21 +/- 0.25 nmol/min/g wt at 90 cmH2O and was strictly parallel with the CPP-CF (coronary perfusion pressure/coronary flow) curve. The autoregulatory range of CF was significantly extended (40-100 cmH2O) under the influence of nifedipine. Hemodynamic effects were accompanied by significant changes in nitrite outflow in all groups except for the verapamil group. Nifedipine and diltiazem induced statistically significant increases of nitrite outflow in coronary venous effluent, strictly parallel with the CPP-CF curve, from 58% at 120 cmH2O to 190% at 40 cmH2O and from 74% at 120 cmH2O to 166% at 40 cmH2O, respectively. On the contrary, amlodipine induced significant reduction of nitrite outflow which was stronger at the lower value of CPP (44-46% at 40-80 cmH2O), compared to the higher value of CPP (32-37% at 100-120 cmH2O). When L NAME was applied in combination with nifedipine or amlodipine, CF was significantly reduced with parallel changes in nitrite outflow. The results show that Ca2+ channel antagonists (verapamil, diltiazem, nifedipine and amlodipine) strongly influence the coronary endothelial L-arginine/NO system in isolated rat heart leading to the difference in nitrite outflow. PMID- 12087922 TI - In vitro and in vivo equivalence of two oral atenolol tablet formulations. AB - A randomised, cross-over, open study of bioequivalence between two different atenolol (CAS 29122-68-7) tablet formulations is presented. An in vitro comparative study between the two formulations was also performed. Both products meet the USP 23 (United States Pharmacopea) specification. The values of similarity factor (f2) and difference factor (f1) obtained ensure sameness or equivalence of the two dissolution curves. Twenty-four healthy volunteers (male/female) participated in the bioequivalence study. Each treatment was given as a single 100-mg tablet following an overnight fast. Atenolol concentrations in plasma were determined up to 30 h after treatment by HPLC. The pharmacokinetic parameters AUC0-infinity, Cmax and Cmax/AUC0-infinity were tested for bioequivalence after logarithmic transformation of data and ratios of tmax were evaluated nonparametrically. The parametric analysis revealed the following test/reference ratios and their 90% confidence intervals (90% CI): 1.06 (0.99 1.13) for AUC, 1.07 (0.97-1.18) for Cmax, and 0.99 (0.94-1.07) for Cmax/AUC0 infinity. The 90% CI for tmax was 0.91-1.23. All parameters showed bioequivalence between both formulations. A discrete fall in both systolic (SBP) and diastolic (DBP) blood pressure was observed after the drug administration. The fall extent (approximately 11 mmHg in supine position) and the time course of both parameters after the drug administration was similar for both formulations. Minimal values for SBP and DBP were achieved at 6 h after the drug administration for both formulations. Heart rates were also reduced after the administration of both formulations of atenolol in a similar extent (12 b.p.m.) and following a similar time profile (i.e. maximal reductions were observed between 1 and 3 h after the drug administration). It can be concluded that both formulations are equivalent in vitro and in vivo. PMID- 12087924 TI - Studies of piroxicam absorption by oral mucosa. AB - The present study was undertaken to investigate, if the non-steroidal anti inflammatory drug (NSAID) piroxicam (CAS 36322-90-4) Fast-Dissolving Dosage Form (FDDF) can be absorbed in the oral mucosa. Piroxicam FDDF was administrated under the tongue to rats with an oesophagus ligation (OL) to prevent the drug entering the stomach and in turn its absorption by the classic way. A group of sham operated (SO) animals received the same piroxicam FDDF dose. After drug administration, a pharmacokinetic study with serial serum sample extractions at 0, 15, 30, 60 and 120 min was performed. It has been found a prompt increase in serum piroxicam levels in OL-rats, which showed different pharmacokinetics from SO-rats. Areas under curve (AUCs) of OL-rat serum piroxicam levels were higher at 15, 30 and 60 min compared to SO-animals. These results indicate that piroxicam FDDF is absorbed in the rat oral mucosa. Moreover, during the first hour, drug absorption by oral mucosa rendered higher piroxicam levels than gastric absorption. PMID- 12087923 TI - Prospective controlled trial of two nifedipine extended release formulations in the treatment of essential hypertension. AB - The aim of this study was to assess the pharmacodynamic equivalence of two different slow-release formulations of nifedipine (CAS-21829-25-4). In a prospective, controlled, double-blind clinical trial, 42 patients with essential hypertension (sitting diastolic blood pressure (DBP) 95-114 mmHg) underwent an initial washout, drug free period of 2 weeks, after which they were randomized to receive either 30 mg of nifedipine in the test preparation "XL" (Nifecard) or 30 mg of nifedipine in a reference formulation, "LA", during six weeks. The response to treatment was assessed by measuring the blood pressure (BP) every two weeks (standard office mercury sphygmomanometry) and by 24-h ambulatory blood pressure monitoring (ABPM). Of the 42 included patients 36 (85.5%) completed the trial: 19 on "XL" and 17 on "LA". After 2 weeks of therapy the DBP decreased by about 11% ( 13.4 mmHg) and 10% (-9.5%), respectively, after 4 weeks the mean decrease versus the end of the placebo period reached about 14% (-15.5 mmHg) and 11% (-13 mmHg), and at the end of the trial the DBPs were lower by about 13% (-14.5 mmHg) in both groups. In all these measurements the within group differences were significant (p < 0.001), while the between groups differences were not (p > 0.05). Quite comparable results were obtained with ABPM, e.g. in the "XL" group the systolic blood pressure at the end of the study was lower by 12.3% (-4.6 mm Hg) and in the "LA" group, by 10.6% (-9.0 mm Hg); p = 0.358. The adverse effects were similar in both groups and they required neither particular interventions nor withdrawal from the study. The drugs under study were comparably effective and well tolerated antihypertensives. PMID- 12087925 TI - Synthesis and evaluation of anticancer activity of some imidazothiazolyl, imidazobenzothiazolyl and dihydroimidazothiazolyl coumarins. AB - A series of 6-(2H-1-benzopyran-2-one-3-yl)imidazo[2,1-b]thiazoles (1), 2-(2H-1 benzopyran-2-one-3-yl)imidazo[2,1-b]benzothiazoles (2) and 3-(2H-1-benzopyran-2 one-3-yl)-5,6-dihydroimidazo[2,1-b]thiazoles (3) have been synthesized and evaluated for anticancer activity in vitro. The compounds showed very good activity against different tumor cell lines. PMID- 12087926 TI - In vitro antiviral activity of aphidicolin and its derivates. Synergistic effects of aphidicolin with other antiviral drugs. AB - Twenty derivatives of aphidicolin were tested against HSV (herpes simplex virus), HCMV (human cytomegalovirus) and adenovirus in vitro. In addition, the antiviral activity of aphidicolin (CAS 38966-21-1) in combination with aciclovir (CAS 59277 89-3) or cidofovir (CAS 113852-37-2) against HSV was determined. The antiviral effects were evaluated using plaque reduction assay in Vero cells or human Foreskin Fibroblasts (HFF) for HSV and HCMV, respectively. Combination indexes were calculated using the method of Chou and Talalay. Two derivatives (K14254 and K14266) that are considered to be prodrugs of aphidicolin were shown to inhibit HCMV and HSV replication comparably to aphidicolin. None of the tested substances inhibited adenovirus replication. Aphidicolin acted synergistically with aciclovir in a 1:1 molar ratio and with cidofovir in different ratios. Aphidicolin and its two antiviral active derivatives might represent useful additional tools for antiviral therapy of HSV and HCMV infections, especially in combination with clinically used drugs. PMID- 12087927 TI - Effect of chloroquine on glucose metabolism. AB - The long- and short-term effects of chloroquine (CAS 54-05-7) on glucose metabolism in rats were assessed. The long-term chronic chloroquine administration (5 and 10 mg/kg b.w. 6 days a week for 6 months) caused a decrease in serum glucose, insulin, calcium, potassium and protein levels, while the glucagon level increased. The short-term acute effect of chloroquine administration (10 mg/kg b.w. 6 days for one week) caused an improvement in glucose tolerance as shown by the decrease in glucose and insulin curves after an oral glucose tolerance test. This was accompanied by an increase in insulin activity, corrected insulin response, and glucose tolerance parameter and a decrease in glucose and insulin areas. Lactate dehydrogenase and glucose-6 phosphate dehydrogenase activities were increased, too, indicating an increase which provides the needed energy for overcoming the injurious effect of chloroquine. PMID- 12087928 TI - Bioequivalence evaluation of a fixed combination of chloroquine and proguanil in a capsule formulation versus a standard medication. AB - To assess the bioequivalence between a test capsule with a fixed combination of chloroquine (CAS 54-05-7) and proguanil (CAS 500-92-5), and chloroquine and proguanil administered as separate tablets, an open two-sequence, two-period cross-over randomized study was performed in twelve healthy volunteers who received a single oral dose of 100 mg chloroquine and 200 mg proguanil either in the form of one capsule or the reference tablets. Biological samples (plasma, whole blood and erythrocytes) were collected up to 43 days after drug administration. The parent drugs and their main metabolites were analyzed using high performance liquid chromatography assay. Bioequivalence was assessed for whole blood and plasma AUC and Cmax of chloroquine, proguanil, cycloguanil and 4 chlorophenylbiguanide. Bioequivalence in erythrocytes was also established except for Cmax of chloroquine. While the differences for Cmax of chloroquine in erythrocytes may be related to technical problems during the erythrocyte sampling procedure (contamination with leukocytes), bioequivalence can be concluded from the plasma concentration data. Therefore, the use of a single capsule instead of one chloroquine tablet and two proguanil tablets daily can be proposed in order to increase the prophylactic compliance without decreasing the prophylactic efficacy. PMID- 12087929 TI - [What is the use of the National Counsel of Universities?]. PMID- 12087930 TI - [Cryopreservation of ovarian tissue]. AB - Ovarian tissue cryopreservation (OTCP) is a new procedure of medically assisted procreation, still at the experimental stage, whose primary aim is to store female gametes as sperm cryopreservation permits to do for male gametes. Ovarian tissue is removed very simply by laparoscopy. It survives well to freezing if the medium contains a cryoprotective agent and the rate of freezing is slow. In contrast, thawing must be rapid. There are three processes for the utilization of ovarian tissue after thawing. In vitro maturation and xenografting remain impossible for technical and ethical reasons. Autologous transplantation (orthotopic or heterotopic) of the tissue is therefore the only foreseeable method over the short term. Indications for OTCP must remain rare as long as no pregnancy has been obtained in human. At the present time, only female patients who would inevitably suffer the loss of their fertility should be able to take advantage of OTCP. Basically, this would mean women subjected to castrating anticancer therapy. It would seem reasonable to set the age limit at 35-years for carrying out OTCP. Lastly, female patients should be clearly informed that the method is still at the research stage, and in France samples must be taken in accordance with the laws governing clinical research. PMID- 12087931 TI - [The role of surgery in the treatment of prolactinomas]. AB - TECHNIQUE: Almost all prolactinomas are operated through a transsphenoidal route, even in case of significant suprasellar extension or intracavernous invasion. Since 1996, we use the only endonasal route which is easier, quicker, less haemorrhagic and less aggressive than the sublabial one. If the removal of a large macroadenoma cannot be completed (50-70% of cases), a second procedure, usually transsphenoidal, is performed a few weeks later. INDICATIONS: Microprolactinomas: usually surgery is proposed as soon as medical treatment is not well tolerated or if the patient wishes to be pregnant. Macroprolactinomas: bromocriptine or quinagolide are sometimes prescribed in a first stage. Surgery beeing indicated in case of inefficacy or intolerance of the medical treatment, or in emergency due to an acute visual pathways compression (tumoral necrosis). Giant adenomas (= 30-40 mm): usually dopaminergic agonists allow a tumoral volume reduction and assure a limitation of visual risks due to rapid decompression of visual pathways. Exceptionally, medical treatment leads to a complete regression of the adenoma. RESULTS: Morbidity of transsphenoidal procedures: insipidus diabetes: transient 5-15%, definitive: 1-2%; septal perforation: 3-5%; rhinorrhea: 5%; visual aggravation: 2%. The most frequent complaint for patients operated through sublabial route is gum and dental pain, non-existent with endonasal procedures. Operative mortality: 0-1.7% (0% in our series). In men, normalization of prolactinemia is linked to the tumoral volume: 90-100% in case of microprolactinoma, 30-35% for enclosed macroprolactinomas, 0-5% for invasive macroprolactinomas. Conventional, conformational or stereotactic radiotherapy can be useful in case of contraindication to surgery and failure of medical treatment. PMID- 12087932 TI - [Compliance with hormone replacement therapy in menopause: effect of an original education program. The COMET study]. AB - OBJECTIVE: To study the impact of an original education program on compliance to hormone replacement therapy (HRT) in post-menopausal women. METHODS: Data were obtained from 1,192 post-menopausal women (age: 53 +/- 5 years) included in the study with an onset less than six months: E2 gel (n = 791) or patch (n = 401) + progestins, and randomized in either educational program (Ep = 600) or regular verbal counselling (VC = 592). A patient is considered bad compliant with HRT when she prematurely stopped the study, whatever the reason of the cessation. The groups EP and VC are homogeneous for the age distribution, the HRT regimen, the date of onset and the climateric symptoms scores. RESULTS: There is a significant difference between EP and VC groups in compliance, respectively 86 and 81% (p < 0.027). This difference is partially explained by the significant improvement in the patch subgroups (EP: 86% versus VC: 77%, p = 0.028). The trend observed in the gel group is not significant (EP: 85% and VC: 82%) due to the high level of compliance usually noticed in women treated with the gel and the verbal counselling. The significative difference in the compliance observed during the verbal counselling between the continuous (76%) and the sequential (86%, p = 0.001) HRT regimen disappears with the educational program (continuous: 84% and sequential: 87%, NS). CONCLUSION: This first large study on the HRT compliance in France, using a patient educational material, validates its efficacy to reach a better compliance than with regular verbal counselling. Even though in the daily practice, about 40% of the patients discontinue the treatment during the first year, 81% of the women followed in this study continue to receive the HRT after nine months of use. The original educational program of the COMET study improves significatively this high compliance (+ 5% from the verbal counselling). Thus, the combination of the doctor verbal counselling and an educational material is desirable to obtain a good compliance with HRT. PMID- 12087933 TI - [Idiopathic granulomatous mastitis: apropos of 8 cases and review of the literature]. AB - AIM OF THE STUDY: The authors report eight cases of idiopathic granulomatous mastitis in which the epidemiological, clinical, diagnostic, therapeutic and prognostic features are discussed. MATERIAL AND METHODS: Retrospective study of eight cases registered among 2,000 breast anatomopathological study during 5 years period. RESULTS: The mead age was 44.3 years. Breast lump measuring 2.5 to 15 cm in size was noted at clinical examination. Mammography noted nodular opacities and ultrasonography showed hypoechoic nodules. Histological findings were consistent on granulomatous inflammation centred on mammary lobules. The inflammatory infiltrate was including leukocytes, lymphocytes, plasma cells as well as epithelioid and giant cells without caseous necrosis in the granuloma. DISCUSSION AND CONCLUSION: Idiopathic granulomatous mastitis is a rare entity. This entity can clinically mimic other breast mastitis disease. Diagnosis is assessed by histological analysis This disease can be treated with drugs and surgical excision of the lump. The prognostic of this chronic disease is favourable. PMID- 12087934 TI - [An atypical Demons-Meigs syndrome]. AB - Meigs'syndrome is defined as a hydrothorax with ascites and benign ovarian tumor, both of which resolve on removal of the tumor. Pseudo-Meigs'syndrome or atypical Meigs'syndrome occurs when a pelvic mass other than an ovarian fibroma is present with hydrothorax and ascites. Both these syndromes should be nevertheless considered in women who present hydrothorax and ascites. This case concerns a 50 year-old woman who presented a pseudo-Meigs'syndrome (Hydrothorax and uterine leiomyoma). The distinction between these two syndrome is useless, because tumour removal is the only treatment. PMID- 12087935 TI - [Which psychological support for couples seeking medical assistance for procreation?]. AB - OBJECTIVES: The objective of our qualitative study was to define modalities of psychological support to be offered to couples seeking medically assisted procreation. MATERIAL AND METHODS: Forty couples participated in a semi structured videotaped interview, which touched on themes such as personal and family histories, the couple's relationship, etc. We focused on the "narrative mobility", that is the way in which the couples transmit their personal and family history during the interview and the interviewer's impression that he may or may not share this with the couples. RESULTS: Observed differences in narrative mobility led us to distinguish three groups of couples and to propose various types of psychological support. DISCUSSION AND CONCLUSION: This exploratory study, with its interest for the narrative mobility, concerns the couples' capacity to stand back from their own story as responsible interlocutors. We made the hypothesis that this capacity is linked to their capacity to handle their emotional stress, to act as partners to the medical team and to prepare themselves for their future parenthood. PMID- 12087937 TI - [MRI and pelvic prolapse]. AB - Static and dynamic pelvic IRM is nowadays part of the diagnostic work up of pelvic prolapse. It is a second line examination, diagnosis being made primarily on the basis of findings at physical pelvic examination. Advantages of MR imaging include lack of ionising radiation, simplicity, multiplanar imaging capability, and possibility to explore at the same time the three compartments and the pelvic floor. The dynamic MR Imaging can be coupled with a clinical examination at the magnet. Imaging is useful in patients in whom findings at physical examination are equivocal. Dynamic MR imaging can be useful in evaluating pelvic organ prolapse. It allows detection of hidden prolapses and assessment of muscle trophycity. Post-operatively it allows assessment of surgical results and failures or recurrences. PMID- 12087936 TI - [Male infertility and microdeletions of the Y chromosome]. AB - It is estimated that about 10% of men suffer from male infertility. Male infertility is associated with a reduction in the quantity, reduced mobility or abnormal morphology of sperm. In about 50-60% of cases the etiology can be identified. When the cause is unknown, it is referred to as idiopathic infertility. A genetic cause is suspected in some of the latter cases since chromosome anomalies and familial forms of male infertility have been reported. Three different regions of the Y chromosome, termed AZFa, AZFb and AZFc are recurrently deleted in about 15% of cases of idiopathic azoospermia or severe oilgozoospermia. AZFc deletions form the majority of these deletions. The presence of a Y microdeletion does not seem to alter the fertilisation of the oocyte or the development of the embryo. However, if the child is a boy, he will inherit the deletion from his father and will most likely be infertile when he is an adult. In the absence of any other information concerning an association between Y chromosome microdeletions and other development anomalies of the child, in genetic counselling the principal risk for male offspring appears to be infertility. PMID- 12087938 TI - [Progestogens, progesterone, coagulation and vascular tone]. AB - Epidemiological studies suggest that estrogens may have opposite effects on vessels: estrogens used for contraception are known to increase both arterial and venous risk, while hormone replacement therapy could reduce the risk of cardiovascular disease. In both situations, estrogens are associated with progestogens. Progestogens are rarely used alone, thus the effect of progestogens on haemostasis or vessel wall is unclear. Data can be obtained from studies using progestogens alone or from studies comparing unopposed estrogens to combined estrogen-progestogen therapy. Progestogens alone have few effect on the haemostatic system. In combined therapy used for contraception, progestogens modify the effects of estrogens on haemostasis and endothelium: the overall effect, including modifications of coagulation factors and inhibitors could a prothrombogen trend, mainly for third generation progestogens. Unopposed estrogens are also rarely used for post menopausal hormone replacement therapy (HRT). Experimental studies have shown that progestogens are able to inhibit the beneficial effect of estrogens. Two mechanisms have been suggested: first, progestogens may reduce the endothelium-dependent vasodilatator action of estrogens. Another explanation concerns the neointimal proliferation leading to atherosclerosis: Estradiol are known to reduce this proliferation. Progestogens could reduce the protective effect of estrogens. These pharmacological effect of progestogens must be taken in account to interpret the negative results of HERS study that failed to demonstrate a cardiovascular benefit of estrogens plus progestin therapy in postmenopausal women. PMID- 12087939 TI - [Third trimester amniocentesis for fetal karyotyping: why so late?]. PMID- 12087940 TI - [Third trimester amniocentesis for fetal karyotyping: should we forget it?]. PMID- 12087941 TI - [Management of mucocutaneous herpes in immunocompetent patients (excluding ocular manifestations) (November 2001). Long version]. PMID- 12087942 TI - [In response to the article by M. Akotionga et al. ]. PMID- 12087943 TI - [In response to the article by M. Laghzaoui Boukaidi et al. ]. PMID- 12087944 TI - [In response to the article by C. Poncelet et al. ]. PMID- 12087945 TI - [In response to the article by P. Merviel et al. ]. PMID- 12087946 TI - Intraoperative identification of recurrent laryngeal nerves in thyroid surgery by electrical stimulation. AB - INTRODUCTION: Iatrogenic injury of the recurrent laryngeal nerve and its prevention is one of the controversial issues in thyroid surgery. The incidence of this operative complication ranges from 0.4% to 3.9%. PATIENTS AND METHODS: The authors analysed the results of the identification of 114 nerves in 60 patients who underwent thyroid surgery between July 9, 1999 and December 19, 2000 in the 3rd Surgical Clinic at the Department of Clinical Surgery and in the Department of Ear Nose and Throat Diseases of the Medical University in Plovdiv, Bulgaria. Nerve integrity monitor Neurosign 100 and specially designed EMG electrodes (Magstim Company Ltd., Wales, UK) were used. The patients were examined pre- and postoperatively for vocal cord paralysis by otolarygologist. RESULTS: In all patients electrical stimulation of the recurrent laryngeal nerve was successfully used for identification of the nerve and evaluation of its integrity during and at the end of the operation. No postoperative recurrent laryngeal nerve deficit was detected clinically. CONCLUSIONS: The results indicate that the identification of the recurrent laryngeal nerve by electromyography is a safe, effective and simple method for electrophysiologic monitoring during thyroid surgery. This method allows assessment of the neural integrity at the end of the procedure. PMID- 12087948 TI - Endonasal surgery in mucopyocele of the paranasal sinuses. AB - INTRODUCTION: Mucopyocele of the paranasal sinuses generally develops as complication in patients with chronic inflammatory diseases of the paranasal sinuses, but could also be secondary to trauma, tumor or surgical manipulation. Surgery is the definitive method of cure. AIM: The aim of the study was to assess the endonasal microsurgical marsupialization in treatment of mucopyocele. MATERIAL AND METHODS: Five patients (3 females and 2 males) with mucopyocele of the frontal and/or ethmoidal sinuses were treated in the Department of Ear, Nose, and Throat Diseases at the Medical University in Plovdiv for the period 1996 1999. The process location was exactly defined by high-resolution computed tomography. Endonasal microendoscopic anterior ethmoidectomy with marsupialization was performed. In two patients (frontal sinus mucocele) the endonasal approach was combined with an extranasal approach keeping the functional principles. RESULTS: The control endoscopic and computer tomographic examinations did not show recurrence of the disease. CONCLUSION: Surgical approach in mucocele of the sinuses should be based on the findings of accurate computed tomography, rather than performed as a routine technique. This allows the operation to be particularized, trauma and duration of the operation reduced and function of the sinunasal system preserved. PMID- 12087947 TI - Method of anaesthesia in thyroid surgery with intraoperative monitoring of the recurrent laryngeal nerves. AB - INTRODUCTION: The incidence of injury of the recurrent laryngeal nerve (RLN) in various types of surgery is reported to be as high as 12%. By monitoring the recurrent laryngeal nerve intraoperatively we can identify it and preserve it even in atypical positions or in operations for processes involving the nerve. These types of surgery give rise to some specific problems facing the anaesthesiologist and the method of anaesthesia used. AIM: The aim of the study was to assess the efficacy and relevance of an anesthetic method in thyroid surgery with intraoperative monitoring of the RLN. Tasks: 1. To determine the level of myorelaxation in anaesthetic patients by intraoperative neurostimulation of the ulnar nerve. 2. By careful dosing of the non-depolarizing myorelaxants to create a "window" in the myorelaxation immediately before identification of the RLN that is detected by a neurostimulator. MATERIAL AND METHODS: Forty eight patients subjected to different types of thyroid surgery were given inhalational anaesthesia with supplementation of non-depolarizing myorelaxants. The level of relaxation was constantly monitored by neurostimulation of the left ulnar nerve. This peripheral stimulation allows creating a "window" in the myorelaxation just before the identification of the recurrent laryngeal nerve. RESULTS: There were no perioperative complications in all patients as to both surgery and anesthesia; postoperative comfort was achieved. The recurrent laryngeal nerve was readily identified and its function was preserved in the postoperative period. CONCLUSIONS: The authors consider as a method of choice the general inhalational anaesthesia supplemented with non-depolarizing myorelaxants and permanent intraoperative monitoring of the level of relaxation in surgery with intraoperative monitoring of the cranial nerve and recurrent laryngeal nerve. PMID- 12087950 TI - Selective neck dissection in treatment of node-positive neck cases. AB - Surgical management of patients with cervical metastases of laryngeal and hypolaryngeal cancer has remained a challenge ever since Dr. Crile described the radical neck dissection. The evolution of neck dissection technique confirms this statement. The modification of the radical neck dissection is justified by the severe cosmetic and functional losses related to it and by the fact that despite the comprehensiveness, the recurrences in the neck are still a significant problem. Applying selective neck dissection in node positive cases seems to be justifiable. Expanding the indications for this operation is justified, because in the absence of factors that have destructive effect on fascial compartments of the neck or disrupt the lymphatic flow, such as gross adenopathy or significant extracapsular spread, the principles on which the selective neck dissection is based are still valid. In this paper we report our experience with 10 patients with N1 and N2 who underwent supraomohyoid and lateral neck dissections with follow-up of at least 2 years. PMID- 12087949 TI - Stimulation electromyography as a method of intraoperative localization and identification of the facial nerve during parotidectomy: review of 15 consecutive parotid surgeries. AB - INTRODUCTION: It has been widely recognized that one of the major hazards during operation of the parotid gland, mainly in tumor resection, is the injury of the facial nerve. Facial nerve monitoring has achieved wide application in otologic and neurotologic procedures to help localization and protection of the facal nerve in the temporal bone. MATERIAL AND METHODS: The authors analyze localization and identification of 15 nerves in 15 patients who underwent parotid gland surgery in the Department of Ear Nose and Throat Diseases of the Medical University in Plovdiv from September 15, 2000 to December 15, 2000. Nerve integrity monitor Neurosign 100 and specially designed electromyographic electrodes were used. RESULTS: In all patients stimulation electromyography of the facial nerve was successfully used to locate, identify and evaluate the integrity of the facial nerve during and at the end of the operation. No postoperative facial nerve injury was detected clinically. In two patients with parotid gland cancer the inferior division of the facial nerve was found infiltrated by the cancer growth. CONCLUSION: The results indicate that identification of the facial nerve by electromyography is a safe, effective and simple method of electrophysiologic monitoring during parotid gland surgery. This method allows assessment of the nerve integrity at the end of the operation. PMID- 12087951 TI - A case of aspergillosis of the nose and paranasal sinuses. AB - Until recently aspergillosis of the paranasal sinuses has been considered a rare disease but nowadays it is frequently discussed in the literature. The reason is the nonspecific clinical course of the disease resembling recurrent chronic rhinosinusitis and its difficult histological and radiological verification. Although there have been clinical suspicions of this diseases in quite a few patients presenting with recurrent polyposis of the sinuses associated with aspirin intolerance and bronchial asthma we could prove sinus aspergillosis histologically and computed-tomographically only in one patient. PMID- 12087952 TI - Some characteristics of acute poisoning with psychoactive substances in the period of socioeconomic transition 1990-2000. AB - INTRODUCTION: Although scarce, the data about acute poisonings with psycho-active substances during the period of socioeconomic crisis in Bulgaria indicate the appearance and increase of the use of inhalants, prescribed and over-the-counter medications and the increase of acute poisoning rate with heroin. AIM: To ascertain some characteristics of acute poisonings with psycho-active substances in Plovdiv region from 01.01.1990 to 31.12.2000. MATERIAL AND METHODS: The whole caseload of 571 acute poisonings with psycho-active substances in Plovdiv region (8.8% of the population of the country) between 1990 and 2000, treated in the only Clinic of Toxicology in the region is studied retrospectively. The variables gender, age, type of psycho-active substance (DSM-IV criteria), grade of severity, duration of use, times of poisoning and some differences between alcohol and opioid poisonings are investigated. RESULTS: Acute poisoning with psycho-active substances are 5.43% of all intoxications. The moderate grade of severity (48.5%), alcohol (62.8%) and opioid (15.3%) intoxications, debut poisoning (86.8%) and poisonings occurring in the initial period of use predominate. An increase of acute poisonings with psycho-active substances in the last third of the studied period (1990-2000) is established--severe forms (2.59 times) and recidivistic intoxication (4 times). Alcohol and opioid poisonings predominate among severe intoxications. The hospital stay is relatively short- 1.75 +/- 0.67 and hospital lethality is 1.6%. CONCLUSIONS: 1. The moderate grade of severity, alcohol and opioid intoxications (especially in the severe grade cases), occurrence in the initial period of use and relatively short hospital stay predominate in acute poisonings with psycho-active substances in the period of socioeconomic crisis. Hospital lethality (1.6%) is ascertained only in severe alcohol intoxication. 2. An increase of acute poisonings with psycho-active substances--severe forms, mild forms and recidivistic intoxications is established in the last third (1998-2000) of the studied period. The most expressed increase is ascertained in 2000. 3. Urgent complex measures are necessary to contain the spread of psycho-active substances and acute poisonings with them. PMID- 12087955 TI - Microsurgical technique as a method for prevention of recurrent laryngeal nerve injury in thyroid surgery. Review of seven consecutive cases. AB - INTRODUCTION: Vocal cord paresis or paralysis due to iatrogenic injury of the recurrent laryngeal nerve is one of the main problems in thyroid surgery. Although many procedures have been introduced to prevent the nerves from lesion the incidence of recurrent laryngeal nerve palsy varies between 1.5-14%. The aim of the study was to evaluate the role of the microsurgical technique as a method, which can help the surgeon to identify the recurrent laryngeal nerve in the operative field. MATERIAL AND METHODS: Fourteen nerves in 7 patients were identified with Zeiss multidiscipline universal surgical microscope with a 250-mm ocular lens. All were female patients with mean age of 35 years (Range 16-53) and diagnosis papillary cancer. They were admitted and operated in the Department of Ear, Nose, and Throat Diseases of the Higher Medical Institute in Plovdiv, Bulgaria. RESULTS: In all patients the recurrent laryngeal nerves were successfully identified along their whole length from the inferior thyroid artery to their entering the laryngeal muscles. None of the patients had temporary or permanent vocal cord palsy. CONCLUSION: Microsurgical technique could be a procedure of great value for prevention of the recurrent laryngeal nerves during thyroid surgery, especially in reoperations, thyroid cancer or retrosternal goitre. PMID- 12087953 TI - Fluoroquinolone prophylaxis for bacterial infections in neutropenic patients with hematologic malignancies. AB - INTRODUCTION: The use of fluoroquinolones as prophylaxis for bacterial infections in patients with chemotherapy induced neutropenia is debatable. The present study was conducted to assess the prophylactic efficacy of fluoroquinolones in adult neutropenic patients with hematologic malignancies. MATERIALS AND METHODS: For the period 1994 through 2000 a prospective, randomized trial was conducted at the University Clinic of Hematology (Medical University, Plovdiv). Two groups of patients were included--experimental group, consisting of 36 patients with 41 granulocytopenic episodes and a control group of 34 patients with 41 granulocytopenic episodes. Non parametrical and psi methods were used for statistical analysis. Kaplan-Meier test was employed to determine patients' survival rate. Comparison of survival rates between the two groups was performed using the log-rank test. RESULTS: A statistically significant difference between the two groups was found in the number of infection-free neutropenic episodes (P < 0.001), in infection-related mortality rate (p = 0.001) and mortality rate within 1 month of the onset of infection (p < 0.001). Difference in long term survival rates was not statistically significant (p > 0.05). CONCLUSIONS: Prophylaxis with fluoroquinolones reduced the occurrence of infection in adult neutropenic patients without affecting the long-term survival. Moreover, it is especially beneficial for patients with severe neutropenia and during the active phase of the disease. PMID- 12087954 TI - Penile cancer--surgical treatment of the primary tumor. AB - OBJECTIVE: To present our experience in the surgical treatment of primary tumor in penile cancer based on the accepted oncologic principles and indications for the existing surgical methods of treatment. MATERIAL AND METHODS: A retrospective analysis of 54 patients with histologically proven malignant penile cancer treated surgically at the Urology department of MHAT "St George", Plovdiv between 1975 and April 2001. RESULTS: Fifty patients were treated with 53 operations performed. Penile amputations were significantly more than penile-preserving operations. DISCUSSION: Differentiation and tumor stage are paramount in the choice of surgical strategy for penile cancer. PMID- 12087956 TI - Autoerotic asphyxia (a case report). AB - The authors report the case of a young man, 18 years of age, who died as a result of autoerotic asphyxia. In the practice of forensic medicine such cases are presented as deaths from mechanic asphyxia in which, using various means and devices, the neck is constricted or the external respiratory orifices are obstructed with the purpose of achieving sexual gratification as a result of the induced hypoxia. These cases pose serious difficulties to the forensic medicine expert and the coroner because of the difficulties of distinguishing it as homicide, suicide or accident. The young man was found hanging by the neck, suspended on a leather belt attached on the door of his room. A mirror was placed in front of the door, reflecting him in full height. He was dressed in women's clothing (a skirt, a blouse, bikini) and a brassiere padded to resemble female breasts. Pornographic pictures from newspapers and magazines were spread on the floor. The young man had no known history of mental disorders, but there was evidence for bisexual tendencies and behaviour. On the basis of a comprehensive evaluation of the data it was accepted that the death was accidental and was attributed to autoerotic asphyxia. All the characteristic features for an act of autoerotic asphyxia, described in the forensic medicine literature, were found in this case. Autoerotic asphyxia is almost unknown as a distinctive psychopathological entity in this country's forensic medicine practice. The deficiency of theoretical knowledge in this field, as well as the lack of personal practical experience in most of the legal physicians and pathologists could result in erroneous interpretation of such case. PMID- 12087958 TI - Frontiers in modeling and control of breathing: integration at molecular, cellular, and systems levels. Proceedings of the VIII Oxford Conference. October 11-15, 2000. N. Falmouth, Massachusetts, USA. PMID- 12087957 TI - A patient with a severe chronic airway obstruction and preserved exercise capacity (a case report). AB - We report a patient with chronic obstructive pulmonary disease (COPD) in whom severe lung function disorders are combined with completely preserved exercise capacity. We assessed the exercise capacity of a 44-year-old man (height 155 cm, BMI 19.6 kg.m-2, FEV1%pred. = 30.9%, FRC%pred. = 158%, KCO%pred. = 46.2%, PaO2 = 64.0 mmHg, Medical Research Council Dyspnea scale = 1, Baseline Dyspnea Index = 10) by the 6-minute walking distance test (6MWD) and the symptom-limited cardiopulmonary exercise test (CPET) on a treadmill using the Bruce protocol. The patient was able to walk 667 meters in the test and achieved peak relative oxygen consumption (VO2/kg) of 21.9 mL.min-1.kg-1. We attribute the preserved exercise capacity of the patient to the combined beneficial effect of the following factors: 1. Efficient extraction of the hemoglobin-transported oxygen from the alveoli (P50 = 3.10 kPa). 2. Optimal right ventricle remodelling with mild hypertrophy, without dilatation and congestion. 3. Hypoxic normoxemia without polyglobulia, resulting in good rheologic properties of blood. 4. A preserved locomotory activity of the patient. Such a combination of severe lung function disorders with mildly pronounced dyspnea and preserved exercise capacity supports the concept that the function profile of COPD patients is multidimensional and therefore such patients should have a complete assessment of their disability condition. PMID- 12087960 TI - Abstracts of the 2nd International Conference on Prospects in the Treatment of Rare Diseases. Trieste, Italy. May 23-26, 2000. PMID- 12087959 TI - Proceedings of the Boden Research Conference on Estrogen and Male Health. November 2-3, 2000. Sydney, Australia. PMID- 12087961 TI - How to evaluate and predict the ecologic impact of antibiotics. Proceedings of the 7th Maurice Rapin Colloquium. St Remy de Provence, France. October 26-28, 2000. PMID- 12087962 TI - Reimbursement for advanced practice nurses. PMID- 12087963 TI - Biophysics of membranes. Proceedings of the International Conference on Life Sciences - 2001. September, 22-26, 2001. Gozd Martuljek, Slovenia. PMID- 12087964 TI - [French Society of Perinatal Medicine, XXXI meeting. Lille, France, 25-26 October 2001. Abstracts]. PMID- 12087965 TI - Systemic medications: clinical significance in periodontics. AB - Systemic medications are of value as adjuncts to periodontal therapy. These medications can be divided into two major categories: antibiotics and agents for host modulation. Antibiotics have been shown to be valuable adjuncts in specialized types of periodontal disease, such as localized and generalized aggressive periodontitis, and of possible value in severe chronic periodontitis. Antibiotics have been studied individually, in combination and in sequential therapy. Host modulators include Periostat, non-steroidal anti-inflammatory agents, alendronate (Fosamax), hormone replacement therapy and anti-arthritic medications. These agents produce their beneficial effects by a variety of mechanisms of action, including inhibition of matrix metalloproteinases, inhibition of prostaglandin production, stimulation of osteoblasts, inhibition of osteoclasts, and other anti-inflammatory mechanisms of action. PMID- 12087966 TI - Bilateral renal oncocytosis with renal failure. PMID- 12087967 TI - Institutional consultations in surgical pathology: how should diagnostic disagreements be handled? PMID- 12087968 TI - Liability and payment issues in the selection of pathology assays. PMID- 12087969 TI - Pathologic quiz case: cystic lesion of the posterobasal portion of the heart in a 68-year old woman. PMID- 12087970 TI - Pathologic quiz case: pulmonary infiltrates with characteristic light and electron microscopic features. PMID- 12087971 TI - Pathologic quiz case: multiple pulmonary nodules in a patient with stage IV, diffuse large B-cell lymphoma. PMID- 12087972 TI - Pathologic quiz case: recurrent spontaneous pneumothorax in an industrial worker. PMID- 12087973 TI - Pathologic quiz case: a 35-year old woman with a history of arrhythmia and liver failure. PMID- 12087974 TI - Urinary gems: acyclovir crystalluria. PMID- 12087975 TI - Plasma cell crystalline inclusions. PMID- 12087976 TI - Biomedical journals in Croatia. PMID- 12087977 TI - The oldest Croatian medical journal. PMID- 12087978 TI - Proceedings of the 10th Symposium on Growth Hormone and Related Factors. Tokyo, Japan. February 14, 1998. PMID- 12087979 TI - Pet nutrition coming of age. Proceedings of the Waltham International Symposium. August 6-7, 2001. Vancouver, Canada. PMID- 12087980 TI - Public fear of dioxins from modern municipal waste incinerators is not justified. PMID- 12087981 TI - Bad air and birth defects. PMID- 12087982 TI - Poor environment creates wealth of problems. PMID- 12087983 TI - Reading, writing, and surfing. PMID- 12087984 TI - NIEHS puts a new face on public service. PMID- 12087985 TI - Results from Russia. The commencement of blood lead testing in kids. PMID- 12087986 TI - It pays to get lead out. The economics of eradication. PMID- 12087988 TI - Canadian Association of General Surgeons Evidence Based Reviews in Surgery. 2. E.A.E.S multicenter prospective randomized trial comparing to-stage vs single stage management of patients with gallstone disease and ductal calculi. PMID- 12087987 TI - The breast milk. Samples provide clues to exposure. PMID- 12087989 TI - Australian clinical nurse supervisors' ethical decision-making style. AB - This paper explores clinical nurse supervisors' ethical styles with regard to supervision in healthcare. Eighty-six registered nurses, all with experience of supervising clinical nurses and students in nursing, completed a specifically designed questionnaire. A qualitative interpretative content analysis identified three core themes: (i) 'Is it safe?'; (ii) 'Is it right?'; and (iii) 'Is it kind?', describing the clinical nurse supervisors' ethical styles. The first core theme 'Is it safe?' covered the supervisors' rules, codes and values that guide their supervisory actions, as well as two sub-themes: (i) empowerment and (ii) integrity. The second core theme 'Is it right?' described the supervisors' responsibility and advocacy as well as the ethical dilemmas experienced in the supervisory process. The third core theme 'Is it kind?' included the supervisors' relationships with patients, professionals and supervisees. The results demonstrate the value of offering a support system, such as clinical supervision, which helps nurses to explore their professional identity for the benefit of the patients. PMID- 12087991 TI - GALEN: breathing life into healthcare capacity building. PMID- 12087990 TI - Versatility and flexibility: attributes of the Critical Incident Technique in nursing research. AB - This article explores the value of using the Critical Incident Technique to capture qualitative data relating to nurses' experiences in three diverse clinical contexts: (i) neonatal intensive care; (ii) palliative care; and (iii) care of the demented elderly. It is suggested that this technique enables the researcher to capture the reality of contemporary nursing practise, and nurses' experiences of that practise. It is also suggested that the versatility and flexibility of the Critical Incident Technique reveals the subtleties and complexities in nurses' experiences of practising nursing in diverse contexts, thereby promoting a deeper understanding of meanings underpinning this practise. PMID- 12087992 TI - How does HIV/AIDS affect African businesses? PMID- 12087993 TI - How to treat without the need to deliver continuous HIV suppression? 9th Conference on Retroviruses and Opportunistic Infections. February 24-28, 2002 - Seattle. PMID- 12087994 TI - Apolipoprotein-E genotype and the risk of developing cholelithiasis following bariatric surgery: a clue to prevention of routine prophylactic cholecystectomy. AB - BACKGROUND: Obesity and especially rapid weight loss following bariatric surgery are known risk factors for cholelithiasis. Since the risk may be high, prophylactic cholecystectomy has been advocated. Apolipoprotein (Apo) E, an important carrier protein in cholesterol metabolism and trafficking, is believed to play a role in gallstone pathogenesis. In particular, the Apo E4 allele has been suggested to be associated with cholesterol cholelithiasis. The aim of this study was to assess the incidence of postoperative cholelithiasis in our patient population and to determine a possible correlation with the Apo-E genotype. METHODS: 134 morbidly obese patients undergoing gastric restrictive surgery [laparoscopic assisted gastric banding (LAGB) or silastic ring vertical gastroplasty (SRVG)] had abdominal ultrasound before and 6 to 12 months after operation, to determine the presence of gallstones. None of the patients enrolled in the study had gallstones before surgery. They did not have a prophylactic cholecystectomy or receive bile salt treatment. Apo-E genotypes were determined by Polymerase Chain Reaction restriction enzyme analysis. RESULTS: 10 patients (7.5%) developed postoperative cholelithiasis. The incidence of cholelithiasis in each ApoE genotype was: E2/E3--1/20 (5%), E3/E3--3/91 (3%), E3/E4--6/21 (29%), and E4/E4--0/2. ApoE allele frequencies in the study population were identical to those of a healthy control population. The mean BMI dropped from 43.6 to 29.4 kg/m2. CONCLUSIONS: The occurrence of postoperative gallstones was low in our population. However, in subjects with the Apo-E3/E4 genotype, the incidence is of practical significance. These data suggest that Apo-E genotyping may be useful in selecting patients for gallstone prevention (surgical or medical) when undergoing bariatric surgery. Further testing in larger patient populations may be able to give more definite guidelines in the future. PMID- 12087995 TI - From the Centers for Disease Control and Prevention. Occupational exposures to air contaminants at the World Trade Center disaster site--New York, September October 2001. PMID- 12087996 TI - From the Centers for Disease Control and Prevention. Rabies in a beaver--Florida, 2001. PMID- 12087997 TI - From the Centers for Disease Control and Prevention. Outbreak of acute gastroenteritis associated with Norwalk-like viruses among British military personnel--Afghanistan, May 2002. PMID- 12087998 TI - JAMA patient page. Intensive care units. PMID- 12087999 TI - HPV E6 antisense induces apoptosis in CaSki cells via suppression of E6 splicing. AB - Cervical cancer is known to be highly associated with viral oncogene E6 and E7 of human papilloma virus. Down-regulation of oncogene expression by antisense-based gene therapy has been extensively studied. To investigate the effect of HPV 16 E6 antisense nucleic acid (AS) on cervical cancer cells, human cervical cancer cell lines, CaSki and SiHa cells harboring HPV 16 genome were transfected with plasmid containing E6(AS). The decreased viability and the apoptotic morphology were observed in E6(AS)-transfected cervical cancer cell lines. By 6 h after transfection, inhibition of E6 splicing, rapid upregulations of p53 and a p53 responsive protein, GADD45, were displayed in E6(AS)-transfected CaSki cells. Furthermore, E6(AS) induced loss of mitochondrial transmembrane potential, release of mitochondrial cytochrome c into the cytoplasm, and subsequent activation of caspase-9 and caspase-3. These results indicate that HPV 16 E6(AS) induces apoptosis in CaSki cells via upregulation of p53 and release of cytochrome c into cytoplasm, consequently activating procaspase-9 and procaspase 3. PMID- 12088000 TI - Next: twin tube babies? PMID- 12088001 TI - The political impact of scientific cooperation on nations in conflict: an overview. PMID- 12088002 TI - The role of scientists in normalizing U.S.-China relations: 1965-1979. PMID- 12088003 TI - Scientific cooperation and conflict management in U.S.-China relations from 1978 to the present. PMID- 12088004 TI - Nuclear cooperation in South America: the role of scientists in the Argentine Brazilian rapprochement. PMID- 12088005 TI - Scientific cooperation in agriculture and medical research as a means for normalizing relations between Egypt and Israel. PMID- 12088006 TI - The influence of Palestinian-Israeli academic encounters on the peace process. PMID- 12088007 TI - The role of the Weizmann Institute of Science in normalizing Israeli-German relations. PMID- 12088008 TI - The impact of pugwash on the debates over chemical and biological weapons. PMID- 12088009 TI - International conflicts over environment: scientists' roles and opportunities. PMID- 12088010 TI - Scientific cooperation as a bridge across the Cold War divide: the case of the International Institute for Applied Systems Analysis (IIASA). PMID- 12088011 TI - The role of seismologists in debates over the Comprehensive Test Ban Treaty. PMID- 12088012 TI - Scientific cooperation, state conflict: the roles of scientists in mitigating international discord. Preface. PMID- 12088013 TI - Drug therapy in human and experimental transmissible spongiform encephalopathy. AB - During the past 30 years, over 60 different chemical compounds have been used to treat experimental animals infected with transmissible spongiform encephalopathies (TSE), including a wide variety of anti-infectious agents, immunomodulating drugs, and chemicals interacting with the lympho-reticular system. Some compounds achieved a prolongation of the incubation period, but this effect decreased or disappeared when they were administered at or near the onset of symptomatic disease. Recent in vitro and tissue culture studies support earlier speculation about the importance of a chemical structure containing both water-soluble and lipid-soluble components, evidently as a means of interaction with the misfolded membrane-bound 'prion' protein. A number of compounds shown to eliminate the protein (or infectivity) in TSE-infected tissue cultures are the subject of ongoing studies in animals, and are under consideration for human drug trials. As with other recalcitrant infections, combinations of drugs with different modes of action are likely to be necessary for any effective therapy. Also, very recent work in developing antibodies that can neutralize in vitro infection (and, in conjunction with genetic engineering, in vivo infection) has renewed interest in the strategies of both active and passive immunization. PMID- 12088014 TI - Individual state driving restrictions for people with epilepsy in the US. PMID- 12088015 TI - Procerus sign in progressive supranuclear palsy. PMID- 12088016 TI - Quiz page. Tuberous sclerosis. PMID- 12088017 TI - Monoclonal gammopathy of undetermined significance. PMID- 12088018 TI - Monoclonal gammopathy of undetermined significance. PMID- 12088019 TI - Bisphosphonates and osteoporosis. PMID- 12088020 TI - Strategies to improve outcomes after renal transplantation. PMID- 12088021 TI - Strategies to improve outcomes after renal transplantation. PMID- 12088022 TI - Strategies to improve outcomes after renal transplantation. PMID- 12088023 TI - Strategies to improve outcomes after renal transplantation. PMID- 12088024 TI - Pharmacotherapy for obesity. PMID- 12088025 TI - Pharmacotherapy for obesity. PMID- 12088026 TI - Protecting research subjects. PMID- 12088027 TI - Protecting research subjects. PMID- 12088028 TI - Effect of clinorotation on in vitro cultured explants of Mentha piperita L. AB - An in vitro culture system was used to study the influence of gravity on axillary shoot formation and adventitious root regeneration in Mentha piperita L. The direction of the gravity vector was altered by displacing stem node explants in different orientations. Also, microgravity conditions were simulated by rotating the explants on a horizontal clinostat so that the main axis of nodes was either parallel (Cpa) or perpendicular to the clinostat axis (Ccp and Ccf, centripetally and centrifugally oriented, respectively). Mint nodes were cultured on solidified Linsmaier and Skoog's medium [Physiol. Plant. 18 (1965) 100] adding a filter sterilized aqueous solution of 2 mg/l benzyladenine (BA) in half of the cultures. The proliferation of axillary shoots as well as adventitious root formation were not affected by altering upright explant orientation. On the contrary clinorotation was able to modify plantlet development. In absence of BA, leaf width was hindered by Cpa treatment and penultimate internode length was enhanced by Ccp. Furthermore, a negative effect of Cpa treatment was observed in root length parameter, while Ccp increased the root number both in absence and in presence of BA. An effect strictly connected to clinorotation in presence of BA was the occurrence of hyperhydricity. Moreover, explants under clinorotation treatments switched their gravitropic response modifying shoot curvature. PMID- 12088029 TI - Machine vision guided sensor positioning system for leaf temperature assessment. AB - A sensor positioning system was developed for monitoring plants' well-being using a non-contact sensor. Image processing algorithms were developed to identify a target region on a plant leaf. A novel algorithm to recover view depth was developed by using a camera equipped with a computer-controlled zoom lens. The methodology has improved depth recovery resolution over a conventional monocular imaging technique. An algorithm was also developed to find a maximum enclosed circle on a leaf surface so the conical field-of-view of an infrared temperature sensor could be filled by the target without peripheral noise. The center of the enclosed circle and the estimated depth were used to define the sensor 3-D location for accurate plant temperature measurement. PMID- 12088030 TI - Possible involvement of the membrane potential in the gravitactic orientation of Euglena gracilis. AB - Euglena gracilis, a unicellular photosynthetic flagellate, uses light and gravity as environmental hints to reach and stay in regions optimal for growth and reproduction. The current model of gravitaxis (the orientation with respect to the earth's gravitational field) is based on the specific density difference between cell body and medium. The resulting sedimentation of the cell body applies a force to the lower membrane. This force activates mechano-sensitive ion channels. The resulting ion flux changes the membrane potential, which in turn triggers reorientational movements of the trailing flagellum. One possibility for recording the predicted membrane potential changes during reorientation is the use of potential-sensitive dyes, such as Oxonol VI. The absorption changes of the dye indicating potential changes were recorded with a custom-made photometer, which allows a high precision measurement with a high temporal resolution. After a gravitactic stimulation, a short period of hyperpolarization was detected, followed by a massive depolarization of the cell. The membrane potential returned to initial values after a period of approximately 200 s. Parallel measurements of the precision of orientation and the membrane potential showed a close relationship between both phenomena. The obtained results support the current model of gravitaxis of Euglena gracilis. PMID- 12088031 TI - The growth of tomato (Lycopersicon esculentum Mill.) hypocotyls in the light and in darkness differentially involves auxin. AB - Light and auxin antagonistically regulate hypocotyl elongation. We have investigated the physiological interactions of light and auxin in the control of tomato (Lycopersicon esculentum Mill.) hypocotyl elongation by studying the auxin insensitive mutant diageotropica (dgt). The length of the hypocotyls of the dgt mutant is significantly reduced when compared to the wild type line Ailsa Craig (AC) in the dark and under red light, but not under the other light conditions tested, indicating that auxin sensitivity is involved in the elongation of hypocotyls only in these conditions. Similarly, the auxin transport inhibitor naphthylphthalamic [correction of naphtylphtalamic] acid (NPA) differentially affects elongation of dark- or light-grown hypocotyls of the MoneyMaker (MM) tomato wild type. Using different photomorphogenic mutants, we demonstrate that at least phytochrome A, phytochrome B1 and, to a much lesser extent [correction of extend], cryptochrome 1, are necessary for a switch from an auxin transport dependent elongation of hypocotyls in the dark to an auxin transport-independent elongation in the light. Interestingly, the dgt mutant and NPA-treated seedlings exhibit a looped phenotype only under red light, indicating that the negative gravitropism of hypocotyls also differentially involves auxin in the various light conditions. PMID- 12088034 TI - Invariants and labels in Lie-Poisson systems. AB - Reduction is a process that uses symmetry to lower the order of a Hamiltonian system. The new variables in the reduced picture are often not canonical: there are no clear variables representing positions and momenta, and the Poisson bracket obtained is not of the canonical type. Specifically, we give two examples that give rise to brackets of the noncanonical Lie-Poisson form: the rigid body and the two-dimensional ideal fluid. From these simple cases, we then use the semidirect product extension of algebras to describe more complex physical systems. The Casimir invariants in these systems are examined, and some are shown to be linked to the recovery of information about the configuration of the system. We discuss a case in which the extension is not a semidirect product, namely compressible reduced MHD, and find for this case that the Casimir invariants lend partial information about the configuration of the system. PMID- 12088033 TI - Poly(ethylene glycol) hydrogel microstructures encapsulating living cells. AB - We present an easy and effective method for the encapsulation of cells inside PEG based hydrogel microstructures fabricated using photolithography. High-density arrays of three-dimensional microstructures were created on substrates using this method. Mammalian cells were encapsulated in cylindrical hydrogel microstructures of 600 and 50 micrometers in diameter or in cubic hydrogel structures in microfluidic channels. Reducing lateral dimension of the individual hydrogel microstructure to 50 micrometers allowed us to isolate 1-3 cells per microstructure. Viability assays demonstrated that cells remained viable inside these hydrogels after encapsulation for up to 7 days. PMID- 12088032 TI - Solid state 31phosphorus nuclear magnetic resonance of iron-, manganese-, and copper-containing synthetic hydroxyapatites. AB - The incorporation of micronutrients into synthetic hydroxyapatite (SHA) is proposed for slow release of these nutrients to crops in the National Aeronautics and Space Administration's (NASA's) Advanced Life Support (ALS) program for Lunar or Martian outposts. Solid state 31P nuclear magnetic resonance (NMR) was utilized to examine the paramagnetic effects of Fe3+, Mn2+, and Cu2+ to determine if they were incorporated into the SHA structure. Separate Fe3+, Mn2+, and Cu2+ containing SHA materials along with a transition metal free SHA (pure-SHA) were synthesized using a precipitation method. The proximity (<1 nm) of the transition metals to the 31P nuclei of SHA were apparent when comparing the integrated 31P signal intensities of the pure-SHA (87 arbitrary units g-1) with the Fe-, Mn-, and Cu-SHA materials (37-71 arbitrary units g-1). The lower integrated 31P signal intensities of the Fe-, Mn-, and Cu-SHA materials relative to the pure-SHA suggested that Fe3+, Mn2+, and Cu2+ were incorporated in the SHA structure. Further support for Fe3+, Mn2+, and Cu2+ incorporation was demonstrated by the reduced spin-lattice relaxation constants of the Fe-, Mn-, and Cu-SHA materials (T'=0.075-0.434s) relative to pure-SHA (T1=58.4s). Inversion recovery spectra indicated that Fe3+, Mn2+, and Cu2+ were not homogeneously distributed about the 31P nuclei in the SHA structure. Extraction with diethylene-triamine-penta-acetic acid (DTPA) suggested that between 50 and 80% of the total starting metal concentrations were incorporated in the SHA structure. Iron-, Mn-, and Cu containing SHA are potential slow release sources of Fe, Mn, and Cu in the ALS cropping system. PMID- 12088036 TI - Dynamical spectra and the onset of chaos. PMID- 12088035 TI - From Jupiter's Great Red Spot to the structure of galaxies: statistical mechanics of two-dimensional vortices and stellar systems. AB - The statistical mechanics of two-dimensional vortices and stellar systems both at equilibrium and out of equilibrium are discussed, with emphasis on the analogies (and on the differences) between these two systems. Limitations of statistical theory and problems posed by the long-range nature of the interactions are described in detail. Special attention is devoted to the problem of "incomplete relaxation" and, in the case of stellar systems, to the "gravothermal catastrophe." The relaxation toward equilibrium, possibly restricted to a "maximum entropy bubble," is described with the aid of a maximum entropy production principle (MEPP). The relation with Fokker-Planck equations is made explicit and the structure of the diffusion current analyzed in terms of a pure diffusion compensated by an appropriate friction or a drift. PMID- 12088037 TI - N-body simulations of galaxies and groups of galaxies with the Marseille GRAPE systems. AB - I review the Marseille GRAPE systems and the N-body simulations done with them. First I briefly describe the available hardware and software, their possibilities and their limitations. I then describe work done on interacting galaxies and groups of galaxies. This includes simulations of the formation of ring galaxies, simulations of bar destruction by a massive compact satellite, of merging in compact groups and of the formation of brightest members in clusters of galaxies. PMID- 12088038 TI - On nonlinear dynamics of three-dimensional astrophysical disks. AB - Nonlinear processes concerned with different aspects of nonlinear dynamics of astrophysical disks--structures, flows, turbulence--are reviewed. Special attention is paid to the influence of the three dimentionality of disks on their nonlinear behavior. PMID- 12088039 TI - Satellites as probes of the masses of spiral galaxies. AB - We present atomic hydrogen (HI) observations and analyses of the kinematics of satellite-primary galaxy pairs. Two estimates for the masses of the primaries are available, one from their rotation curves and one from the orbital properties of the satellites. Defining chi as the ratio of these two mass estimates, it is a measure of the presence, or absence, of a significant halo. The chi distribution is presented and the selection effects are discussed. We show that our data, compared with the more numerous pairs identified by Zaritsky et al., have similar distributions for projected separations of less than 200 kpc, even though the selection criteria employed were quite different. Observational biases have a negligible effect; the biased and unbiased distributions are essentially identical. N-body calculations were executed to simulate the dynamical behavior of relatively low mass satellites orbiting primary disk galaxies with and without extended halos. In addition, we made a partially analytical analysis of the behavior of orbits in a logarithmic potential. We find that a "generic" model, characterized by a single disk-halo combination, cannot reproduce the observed P(chi) distribution. However, a simple two-component population of galaxies, composed of not more than 60% with halos and 40% without halos, is successful, if galaxies have dimensions of order 200 kpc. If galaxies are considerably larger with sizes extending to 400 kpc or more, no generic model can describe the full range of the observed P(chi), particularly if the distribution for r(p) < 200 kpc is compared with that for r(p) > 200 kpc. Regardless of the mix of orbital eccentricities, neither pure halo, nor canonical models (disk and halo masses are comparable within the disk radius) will work. A multicomponent approximation can be constructed; the canonical model must be mixed with a small fraction of systems essentially devoid of a massive dark halo. Only by including these complexities can the full range of P(chi) be modeled with any degree of success over all radial extents. We show that dynamical friction cannot be ignored in these explorations and that the average mass of a galaxy is in the range of (1-5) x 10(12) M, with a mass-to-luminosity ratio of at most a few hundred. This is insufficient to close the Universe. PMID- 12088040 TI - Chaos in the centers of galaxies. AB - We compare diverging evolution of a two-component (gas+stars) galactic disk embedded in a "live" halo with that of an identical pure stellar disk. Our modeling supports the conjecture that the growth of central concentration in galaxies dissolves the main family of regular orbits in the stellar bar and assists in the formation of a galactic bulge. PMID- 12088041 TI - Counterrotating galaxies and accretion disks. AB - Theoretical interest in astrophysical disks with counterrotating components of stars and/or gas has been stimulated by recently discovered counterrotating spiral and S0 galaxies. A variety of physical processes can occur in counterrotating disks. We have shown that a strong two-stream instability can occur for one armed (m = 1) tightly-wrapped spiral waves between co and counterrotating stellar components and/or between a corotating stellar component and a counterrotating gaseous component. The instability of counterrotating stellar components has been clearly seen in computer simulations. The unstable two-stream spiral waves can provide an effective viscosity for the gas causing its rapid accretion. Accretion disks consisting of counterrotating gaseous components may exist with an intervening shear layer. Configurations of this type can arise from the accretion of newly supplied counterrotating gas onto an existing corotating gas disk. For example, the gas above the disk midplane can rotate with angular rate +omega(r) while that well below has the same properties but rotates with rate -omega(r). Using the Shakura-Sunyaev alpha turbulence model, we find self-similar solutions where a thin (relative to the full disk thickness) equatorial layer accretes very rapidly, essentially at free-fall speed. This type of accretion flow has now been observed in hydrodynamic simulations. PMID- 12088042 TI - Global spiral patterns in galaxies: complexity and simplicity. AB - A coherent exposition of the density wave theory of galactic spirals is presented in a recent monograph. It is centered on the working hypothesis of quasi stationary spiral structure, a possibility first proposed by Bertil Lindblad. This hypothesis has since been found to be widely applicable in a number of physical contexts, including the explanation of the Hubble classification system and other categorical classes. Direct empirical support of this hypothesis has been provided especially by the regularity of the infrared images frequently observed in a number of galaxies (e.g., NGC 309), and by the observed amplitude modulation along the spiral arms (e.g., M51, M81, and NGC 1300). The present paper is a brief review of this theory with further clarification of the fundamentals and of certain specific issues raised in the literature. On the theoretical side, the likelihood for the validity of this hypothesis has been supported by modal studies. Emphasis is placed on the widely observed coexistence of a single regular structure in the Pop II objects and the more complex irregular structures in the Pop I objects, a contrast first discovered by Zwicky many years ago in the main disk of M51. It is pointed out that, in both barred and nonbarred spirals, this basic phenomenon may be understood by noting the fact that the microscale of the collisionless system of Pop II stars--i.e., the diameter of the epicycle--is typically on the same order of magnitude as the observed spacing between the spiral arms. The spiral pattern is thus a very compact structure which is unlikely to respond readily to internal and external disturbances of moderate magnitudes. This apparent robustness in structure is suggested by its observed regularity despite the impact from the coexisting Pop I objects with their strong irregular turbulent motions. This physical picture supports Oort's conjecture of a limited role for tidal interaction in most of the spiral structure of galaxies. It also enables us to place in proper perspective a controversy between two schools of thought, with different emphasis placed on intrinsic mechanisms and on tidal interaction. It is primarily a matter of applicability, or frequency of occurrence of the different scenarios proposed. There is, as yet, little observational evidence presented in the literature that would support the need of an interpretation of the global spiral structure in the galactic disk in terms of a fast evolving structure recently generated from a featureless initial state through tidal interaction. Dynamically, the likelihood for realizing such a scenario is also estimated to be quite low. The above discussions are placed in the context of a general point of view much advocated in current scientific literature; namely, the need to analyze alternative mathematical models for diverse physical contexts in the study of "complex" systems and to judge separately the merits of each model on the basis of its empirical confirmation in an appropriate physical context. PMID- 12088043 TI - Candidates for abundance gradients at intermediate red-shift clusters. AB - In this work I present evidence for the possible existence of central abundance enhancements in two clusters of galaxies at intermediate red-shifts: Abell 1413 (z approximately 0.14) and Abell 2390 (z approximately 0.23). The results are based on ASCA (Advanced Satellite for Cosmolgy and Astrophysics) data reduction of x-ray spatially resolved spectra. The Fe abundance for A1413 drops from 0.37 +/- 0.08 (in the central 2 arcmin) to 0.20 +/- 0.06 (in the outer 2-6 arcmin), while in A2390 the very central abundance is 0.77 +/- 0.45 (central arcmin) and drops to 0.12 +/- 0.12 in the outer regions. PMID- 12088044 TI - Scaling regimes in the distribution of galaxies. AB - If we treat the galaxies in published redshift catalogues as point sets, we may determine the generalized dimensions of these sets by standard means, outlined here. For galaxy separations up to about 5 Mpc, we find the dimensions of the CfA galaxy set to be about 1.2, with only a modest indication of multifractality. For larger scales, out to about 30 Mpc, there is also good scaling with a dimension of about 1.8. For even larger scales, the data seem too sparse to be conclusive, but we fmd that the dimension is climbing as the scales increase. We report simulations that suggest a rationalization of such measurements, namely that in the intermediate range the scaling behavior is dominated by flat structures (pancakes) and that the results on the smallest scales are a reflection of the formation of density singularities. PMID- 12088045 TI - Recent progress in the study of one-dimensional gravitating systems. AB - We review recent developments in the study of two different interacting gravitational systems: the system of parallel planar mass sheets and the system of concentric spherical mass shells. The approach to equilibrium of a system of parallel planar mass sheets is investigated. Parallels with three-dimensional systems are described. Mass segregation and kinetic energy equipartition in a two component system of planar mass sheets is demonstrated via numerical simulation. The existence of two distinct phases is demonstrated in the system of spherical mass shells. The nature of the transition in the microcanonical, canonical, and grand canonical ensembles is studied both theoretically in terms of mean-field theory and via dynamical simulation. PMID- 12088046 TI - Modeling the time variability of black hole candidates. AB - We present model light curves for accreting black hole candidates (BHC) based on a recently developed model of these sources. According to this model, the observed light curves and aperiodic variability of BHC are due to a series of soft photon injections at random (Poisson) intervals and the stochastic nature of the Comptonization process in converting these soft photons to the observed high energy radiation. The additional assumption of our model is that the Comptonization process takes place in an extended but non-uniform hot plasma corona surrounding the compact object. We compute the corresponding power spectral densities (PSD), autocorrelation functions, time skewness of the light curves, and time lags between the light curves of the sources at different photon energies and compare our results to observation. Our model reproduces the observed light curves well, in that it provides good fits to their overall morphology (as manifest by the autocorrelation and time skewness) and also to their PSDs and time lags, by producing most of the variability power at time scales approximately > a few seconds, while at the same time allowing for shots of a few msec in duration, in accordance with observation. We suggest that refinement of this type of model along with spectral and phase lag information can be used to probe the structure of this class of high energy sources. PMID- 12088047 TI - Stellar oscillons. AB - We study the weakly nonlinear evolution of acoustic instability of a plane parallel polytrope with thermal dissipation in the form of Newton's law of cooling. The most unstable horizontal wavenumbers form a band around zero and this permits the development of a nonlinear pattern theory leading to a complex Ginzburg-Landau equation (CGLE). Numerical solutions for a subcritical, quintic CGLE produce vertically oscillating, localized structures that resemble the oscillons observed in recent experiments of vibrated granular material. PMID- 12088048 TI - From quasars to extraordinary N-body problems. AB - We outline reasoning that led to the current theory of quasars and look at George Contopoulos's place in the long history of the N-body problem. Following Newton we find new exactly soluble N-body problems with multibody forces and give a strange eternally pulsating system that in its other degrees of freedom reaches statistical equilibrium. PMID- 12088049 TI - Chaos in cosmological Hamiltonians. AB - This paper summarizes a numerical investigation which aimed to identify and characterize regular and chaotic behavior in time-dependent Hamiltonians H(r, p, t) = p(2)/2 + V(r, t), with V = R(t)V0(r) or V = V0[R(t)r], where V0 is a polynomial in x, y, and/or z and R(t) proportional to tP is a time-dependent scale factor. When p is not too negative, one can distinguish between regular and chaotic behavior by determining whether an orbit segment exhibits a sensitive dependence on initial conditions. However, chaotic segments in these potentials differ from chaotic segments in time-independent potentials in that a small initial perturbation will usually exhibit a sub- or superexponential growth in time. Although not periodic, regular segments typically exhibit simpler shapes, topologies, and Fourier spectra than do chaotic segments. This distinction between regular and chaotic behavior is not absolute since a single orbit segment can seemingly change from regular to chaotic and vice versa. All these observed phenomena can be understood in terms of a simple theoretical model. PMID- 12088050 TI - Phase space transport in noisy Hamiltonian systems. AB - This paper analyzes the effect of low amplitude friction and noise in accelerating phase space transport in time-independent Hamiltonian systems that exhibit global stochasticity. Numerical experiments reveal that even very weak non-Hamiltonian perturbations can dramatically increase the rate at which an ensemble of orbits penetrates obstructions like cantori or Arnold webs, thus accelerating the approach toward an invariant measure, i.e., a microcanonical population of the accessible phase space region. An investigation of first passage times through cantori leads to three conclusions, namely: (i) that, at least for white noise, the detailed form of the perturbation is unimportant, (ii) that the presence or absence of friction is largely irrelevant, and (iii) that, overall, the amplitude of the response to weak noise scales logarithmically in the amplitude of the noise. PMID- 12088051 TI - Papers by George Contopoulos. PMID- 12088052 TI - Orbital complexity, short-time Lyapunov exponents, and phase space transport in time-independent Hamiltonian systems. AB - This paper compares two alternative characterizations of chaotic orbit segments, one based on the complexity of their Fourier spectra, as probed by the number of frequencies n(k) required to capture a fixed fraction k of the total power, and the other based on the computed values of short-time Lyapunov exponents chi. An analysis of orbit ensembles evolved in several different two- and three dimensional potentials reveals that there is a strong, roughly linear correlation between these alternative characterizations, and that computed distributions of complexities, N[n(k)] and short-time chi, N[chi] often assume similar shapes. This corroborates the intuition that chaotic segments which are especially unstable should have Fourier spectra with particularly broad-band power. It follows that orbital complexities can be used as probes of phase space transport and other related phenomena in the same manner as can short-time Lyapunov exponents. PMID- 12088053 TI - Bifurcations of periodic orbits in axisymmetric scalefree potentials. AB - We study orbits in potentials with central cusps, emphasizing the spheroidal equidensity (SED) potentials generated by mass distributions with spheroidal equidensity surfaces. The most prominent bifurcations are those related to 1:1 and 4:3 resonances between radial motions and motions perpendicular to the central plane. We find that 1:1 resonances can cause the thin tube orbit, as well as the equatorial plane orbit, to become unstable. We concentrate on period tripling bifurcations because they appear to be the least understood. We study them via a class of analytic maps. This study suggests that stable period-three orbits generally arise de novo in stable and unstable pairs via a turning-point bifurcation, and not through a bifurcation from the thin tube at a 120 degree rotation angle. The stable period-three orbits typically have only a short span of existence before becoming unstable to a period-doubling instability through a supercritical pitchfork bifurcation. PMID- 12088054 TI - Irregular period-tripling bifurcations in axisymmetric scalefree potentials. AB - We investigate the phenomenon of irregular bifurcations in axisymmetric scalefree potentials, in which a tube orbit gives rise to a new family of orbits of period 3. A proper understanding of the formation of this irregular family of orbits is important because the family occupies a fair portion of the phase space in models where it occurs. We show that these bifurcations are related to the transition of the rotation number of the phase-space torus through 1/3. This type of bifurcation is closely related to the behavior of nontwist maps. PMID- 12088055 TI - Negative energy modes and gravitational instability of interpenetrating fluids. AB - We study the longitudinal instabilities of two interpenetrating fluids interacting only through gravity. When one of the constituents is of relatively low density, it is possible to have a band of unstable wavenumbers well separated from those involved in the usual Jeans instability. If the initial streaming is large enough, and there is no linear instability, the indefinite sign of the free energy has the possible consequence of explosive interactions between positive and negative energy modes in the nonlinear regime. The effect of dissipation on the negative energy modes is also examined. PMID- 12088056 TI - Gamma irradiation effects and EPR investigation on poly(lactide-co-glycolide) microspheres containing bupivacaine. AB - The effects of gamma radiation on the stability of microspheres made of a polylactide-co-glycolide 50:50 copolymer (PLGA) and loaded with 40% bupivacaine (BU) were studied. The radiolysis mechanisms of BU and BU-loaded microspheres were investigated by using electronic paramagnetic resonance (EPR) analysis. Microspheres were prepared by means of a spray drying method. Gamma Irradiation was carried out in the open, at the dose of 25 kGy, by using a 60Co source. The stability of BU-loaded microspheres was evaluated over a 1-year period on the basis of drug content and dissolution profile. Non-irradiated microspheres were stable over the whole period under consideration. Immediately after irradiation the amount of BU released after 24 h from irradiated microspheres increased from 17 to 25%; in the following 3 months of storage it increased to about 35%, and then it kept constant for 1 year. Radicals generated by BU irradiation were identified by EPR analysis; the sensitivity to gamma radiation of BU was about four times lower than that of PLGA. Furthermore, the EPR spectra of loaded microspheres showed that the relative abundance of BU radicals plus PLGA radicals was proportionate to the electronic fractions of the components; this implies that no spin transfer BU/PLGA had occurred during gamma irradiation. PMID- 12088058 TI - Simple spectrophotometric determination of acyclovir in bulk drug and formulations. AB - A simple and cost effective spectrophotometric method is described for the determination of acyclovir in bulk drug and in formulations. The method is based on the formation of blue coloured chromogen when the drug reacts with Folin Ciocalteu (F-C) reagent in alkaline medium. The coloured species has an absorption maximum at 760 nm and obeys Beer's law in the concentration range 50 450 microg ml(-1). The absorbance was found to increase linearly with increasing concentration of acyclovir, which is corroborated by the calculated correlation coefficient value of 0.9998 (n = 9). The apparent molar absorptivity and Sandell sensitivity were 1.65 x 10(2) l mol(-1) cm(-1) and 1.36 microg cm(-2), respectively. The slope and intercept of the equation of the regression line are 6.87 x 10(-4) and 8.33 x 10(-3), respectively. The limit of detection was 5.68 microg ml(-1) and the limit of quantification was 18.95 microg ml(-1). The proposed method was successfully applied to the determination of acyclovir in pharmaceutical formulations. The reliability of the assay method was established by parallel determination by standard-addition method, and by recovery studies. The results demonstrated and the procedure is at least as accurate, precise and reproducible (RSD < 2%) as the official method, while being simple and less time consuming. A statistical analysis indicated that there was no significant difference between the results obtained by the proposed procedure and those of the official method. PMID- 12088057 TI - Spectrophotometric method for the determination of nifedipine with 4 (methylamino)phenol and potassium dichromate. AB - A new simple, sensitive and reproducible spectrophotometric method for the determination of nifedipine in pure and dosage forms has been proposed. It is based on the reduction of nifedipine with Zn/NNH4Cl, followed by coupling with N methyl-1,4-benzoquinoneimine--the oxidation product of 4-(methylamino)phenol, to give a chromophore which absorbed maximally at 525 nm. The experimental conditions were optimised and Beer's law was obeyed over the concentration range of 5-175 microg ml(-1). The molar absorptivity, detection limit, recovery and RSD were found to be 1.9 x 10(3) l mol(-1) cm(-1), 1.1 microg ml(-1), 99.7-100.5% and 0.3-0.8%, respectively. The proposed method was compared favourably with the official B.P. method. PMID- 12088059 TI - Synthesis and microbiological activity of some N-(2-hydroxy-4 substitutedphenyl)benzamides, phenylacetamides and furamides as the possible metabolites of antimicrobial active benzoxazoles. AB - The synthesis of some N-(2-hydroxy-4-substitutedphenyl)benzamides, phenylacetamides and furamides as the possible metabolites of benzoxazoles (II1 15) was performed in order to determine their in vitro antimicrobial activity against three Gram-positive bacteria, two Gram-negative bacteria and the fungus Candida albicans and their activities were compared with several control drugs. The compounds II11, II12, and II13 were found active at a MIC value of 12.5 microg/ml against the Gram-negative microorganism Pseudomonas aeruginosa. Most of the compounds show antibacterial activity at MIC a value of 25 microg/ml against the Gram-positive bacteria Staphylococcus aureus. For the antifungal activity against C. albicans, compound II10 was found more active than the other derivatives. The antimicrobial activity of some of these benzamides, phenylacetamides (II1 and II10) which are the possible metabolites of benzoxazoles, was also compared to their corresponding cyclic analogues III-IV. Compound II10 possesses two dilutions better antifungal activity than its cyclic analogue, benzoxazole IV, against C. albicans. PMID- 12088060 TI - Synthesis and trichomonacidal activity of perketals and hydroperoxides. AB - Some perketals were synthesized by the Dussault procedure using simple bromides and 2-methoxyprop-2-yl hydroperoxide. Treatment with acetic acid gave the corresponding hydroperoxides. Both perketals and hydroperoxides were tested in vitro as trichomonacidal agents. Most of them exhibited very good activities. The most powerful compound was 2-methoxyprop-2-yl hexadec-l-yl peroxide which exhibited an IC50 value of 0.51 microM being 10 times more effective than the reference compound Metronidazole. PMID- 12088061 TI - Polarographic behaviour of meloxicam and its determination in tablet preparations and spiked plasma. AB - Meloxicam is a new non-steroidal anti-inflammatory drug (NSAID), which has a higher activity cyclooxygenase-2 (COX-2) than against cyclooxygenase-1 (COX-1), with potentially high anti-inflammatory and analgesic action. The voltammetric behaviour of meloxicam was studied using direct current (DC), differential pulse polarography (DPP) and cyclic voltammetry (CV). The influence of several variables (including nature of the buffer, pH, concentration, modulation amplitude, scan rate, drop size, etc.) was examined in DPP method for meloxicam. The best DPP response was obtained in acetate buffer pH 4.88. The peak currents were measured with a static mercury drop electrode at -1.49 V versus Ag/AgCl. Calibration curve for meloxicam was linear at a concentration range from 0.38 to 15.0 microg ml(-1). The method was validated and applied to the determination of meloxicam in tablets, which were in two different dosage forms. A spectrophotometric method reported in the literature was utilized as a comparison method. There were no significant differences between the results obtained by two methods. DPP method is also available and applicable for the determination of mentioned substance in plasma. Mean recovery was 99.20+/-0.37%. It was concluded that the developed method was accurate, sensitive, precise, reproducible and useful for the quality control of meloxicam in pharmaceuticals and spiked plasma. PMID- 12088062 TI - Cytisine derivatives as high affinity nAChR ligands: synthesis and comparative molecular field analysis. AB - A number of new N-substituted cytisine derivatives were prepared and tested, along with similar compounds already described by us and others, as high affinity neuronal acetylcholine receptor ligands. Structure-affinity relationships were discussed in the light of our recently proposed pharmacophore model for nicotinic receptor agonists. The most significant physicochemical interactions modulating the receptor-ligand binding were detected at the three dimensional (3D) level by means of comparative molecular field analysis (CoMFA). The best predictive PLS model was a single-field steric model showing good statistical figures: n = 17, Q2 = 0.717, s(ev) = 0.566, r2 = 0.942, s = 0.275. PMID- 12088063 TI - Synthesis and biological evaluation of a novel pyroglutamyl-modified TRH analogue. AB - The TRH analogue 3, incorporating the (S)-isothiazolidine-1,1-dioxide-3 carboxylic acid (1) moiety in place of the native L-pyroglutamic acid (pGlu) residue, has been synthesized and fully characterized by 1H and 13C NMR. The effects of replacing pGlu with its sulphonamido counterpart on biological activity have been investigated. This peptide, which is significantly stabilized towards hydrolysis by pyroglutamyl peptidase type I (PP I, EC 3.4.19.3), has shown to maintain in vitro prolactin-releasing activity. PMID- 12088064 TI - Synthesis and pharmacological evaluation of some (pyridyl)cyclopropylmethyl amines and their methiodides as nicotinic receptor ligands. AB - A series of 3- and (4-pyridyl)cyclopropylmethyl amines and their quaternary ammonium derivatives have been synthesized; they can be considered as rigid analogues of nicotine. The compounds have been tested on rat cerebral cortex to measure the affinity for the central nicotinic receptor. Only the methiodides show affinity in the micromolar range. The results obtained can provide useful information on the topography of the nicotinic receptor-binding site. PMID- 12088065 TI - Analytical methods for the monitoring of solid phase organic synthesis. AB - Solid phase synthesis (SPS) is a powerful technique to assemble compound libraries in high-throughput parallel and combinatorial synthesis. The widespread applications of these techniques required the development of analytical methods for both structural elucidation and reaction monitoring. This review covers some recently developed techniques for on-bead analyses together with solution-state ones. Particular emphasis is devoted to software and hardware improvements for automated high-throughput analysis. PMID- 12088066 TI - Morphological and histochemical observations of the organic components of ostrich eggshell. AB - The organic component of the avian eggshell can be divided into 3 portions, the shell membranes, the matrix and the cuticle. These have been well characterised in the chicken but little has been published with regard to the ostrich (Struthio camelus). A number of recent studies have indicated that the cause of intra-shell embryonic deaths in the ostrich is similar to intra-shell embryonic deaths that occur in the chicken. These deaths in the chicken are associated with the loss of or damage to the waxy cuticle and other organic components of the eggshell, which is reported to be absent in the ostrich eggshell. In this study, preliminary morphological and histochemical analyses, at the level of the light and electron microscope, have characterised the various organic components of the ostrich eggshell. The results of the histochemical and electron microscopical analyses suggest that there may only be 1 shell membrane in this species, which could play a major role in the limitation of bacterial penetration to the embryonic chamber The shell membrane has a distinct elemental profile as determined by EDS analysis. The matrix is shown to decrease in mesh size from the mammillary layer to the vertical crystal layer. The closer packing of the mesh may indicate the presence of a morphologically discernible termination signal to calcification or the remnants of an evolutionary calcified cuticle. The matrix of the pores may also form a defensive barrier against bacterial invasion, which could be damaged as a result of dipping the eggs before incubation. PMID- 12088067 TI - Another human ear tick case. PMID- 12088068 TI - A congenital defect of the distal forelimb of a cat and of a dog. PMID- 12088069 TI - A preliminary undifferentiated faecal egg count reduction test survey in the Caledon area. AB - During October 1998 a study was performed in the Caledon area to determine the presence of anthelmintic resistance in the southern Western Cape, which falls within the winter rainfall region of South Africa. The study took the form of an undifferentiated faecal egg count reduction test (FECRt) survey. The predominant worm genera of the region are Teladorsagia and Trichostrongylus, but Haemonchus causes occasional outbreaks in sheep. No resistance data for any area in the Western Cape Province are available at present. Resistance (< 90% FECR) was recorded on 73% of the farms included in the study, with 46% of these involving resistance to 1 drench, 36% to 2 drenches and 18% to 3 drenches. No moxidectin resistance was detected when it was administered at the therapeutic dosage of 0.2 mg/kg. The results of this study show clearly that resistance of nematodes to other anthelmintics occurs in the Caledon area. PMID- 12088070 TI - Ixodid ticks on indigenous goats owned by small-scale farmers in four communal grazing areas in South Africa. AB - Indigenous goats belonging to small-scale farmers in 4 communal grazing areas in South Africa were sampled for ixodid ticks during the period September 1991 to May 1993. Three of these areas were in the North West Province (Rietgat, Madinyane and Bethany), and one in Mpumalanga Province (Geluk). No tick control was practised unless requested by the owners. Seven ixodid tick species, of which the majority were immature ticks, were collected from the goats in North West Province. Amblyomma hebraeum was the most numerous of these, followed by Rhipicephalus evertsi evertsi and Rhipicephalus appendiculatus. Substantially more ticks were collected from goats at Rietgat than at the Madinyane and Bethany grazing areas. Five tick species were collected at Geluk and R. evertsi evertsi comprised more than 95 % of the total population. At Rietgat and Geluk A. hebraeum nymphs were present on goats throughout the year, while most R appendiculatus nymphs were collected during September and October 1991 and most adults during January and February 1992. At both Rietgat and Geluk most immature R. evertsi evertsi were collected from spring to late summer, while adults were present throughout the year. PMID- 12088071 TI - Possible application of a nematophagous fungus as a biological control agent of parasitic nematodes on commercial sheep farms in South Africa. AB - Biological control of parasitic nematodes of livestock is currently under development and represents another tool that may be integrated into helminth parasite control strategies. This paper presents a brief introduction to commercial sheep farming in South Africa and currently available nematode parasite control methods. These include the FAMACHA clinical assay, strategies of pasture management, dilution of resistant worm species by introduction of susceptible worms, breed resistant sheep and nutritional supplementation. The purpose of this paper is to outline the principles of biological control using nematophagous fungi and how it may be applied on sheep farms in South Africa. PMID- 12088072 TI - An epidemiological survey demonstrating decline in reproductive efficiency with age and non-seasonality of reproductive parameters in German shepherd bitches in Kenya. AB - An epidemiological survey undertaken in Kenya indicated that 2 previously well established factors, namely decline in reproductive efficiency with age, and non seasonality of canine reproductive parameters, hold true for German shepherd (GSD) bitches in Kenya. Data collection forms were distributed to randomly selected GSD breeders and information so obtained was verified using East African Kennel Club records. Whelping was recorded throughout the year. The litter size varied from 1 to 14 pups per litter with a mean of 6.3 +/- 0.4 SD puppies. Records of 567 whelpings and 3592 puppies were studied. The mean monthly whelping rate was 47.3 +/- 7.2. There was no significant difference in the mean litter size by month (P < 0.05). PMID- 12088073 TI - Neosporosis in a white rhinoceros (Ceratotherium simum) calf. AB - A 16-day-old white rhinoceros calf died suddenly while in excellent condition and showing no obvious previous clinical signs. It was the 9th calf of a mature female kept free-ranging with 11 other rhinoceros as well as various other game species on a 2000 hectare game breeding centre adjacent to the town of Lichtenburg and outlying cattle farmlands in the Northern Province. At post mortem examination, death was ascribed to heart failure. There was marked multifocal to coalescing subacute parasitic myocarditis with numerous protozoan bradyzoite cysts and free tachyzoites present amongst the predominantly round cell inflammatory infiltrate. The coccidian was positively identified as Neospora sp. using both polyclonal and murine monoclonal Neospora caninum antibody immunohistochemistry in the avidin-biotin technique. The parasites stained poorly with Toxoplasma gondii-specific immunoperoxidase staining. Ultrastructurally, a section of a bradyzoite-containing cyst, as well as tachyzoites, were largely but not totally consistent with those described for Neospora caninum. The dam showed no sign of illness. Neosporosis affecting white rhinoceros (Ceratotherium simum) has not previously been reported. A summarised overview of neosporosis from selected publications and a recent review is given. PMID- 12088074 TI - Genetic parameters for resistance to Haemonchus contortus in Merino sheep in South Africa. AB - Resistance to natural infections of Haemonchus contortus (wireworm) was studied in Merino sheep. From February 1995 to July 1996 data were collected from the OTK Merino Stud on the farm Wildebeesfontein near Hendrina in Mpumalanga province. From March 1998 to January 2000 another Merino flock at Irene in Gauteng province was included in the study. In both cases, 50 randomly-chosen animals from each group of lambs weaned were sampled weekly for faecal egg count (FEC) until a mean of 2500 eggs per gram faeces (epg) was reached. At this stage, all lambs in the group were sampled for FEC, packed cell volume (PCV) and live mass in the case of the Wildebeesfontein flock. Animals were monitored from weaning to about 1 year of age. Data from 5 different groups of lambs (583 animals and 1722 records in total) with genetic links were accumulated and analysed. Variance components and resulting heritabilities for logFEC and PCV were estimated using a bivariate animal model with repeated records. The estimated heritabilities were 0.24 +/- 0.02 for logFEC and 0.14 +/- 0.02 for PCV. A strong negative correlation (-0.84 +/- 0.06) existed between the 2 traits. The genetic correlation between PCV and live mass for the Wildebeesfontein flock was low (0.28 +/- 0.09), while that between logFEC and live mass was negative, although not significantly different from zero (-0.13 +/- 0.09). These results suggest that FEC can be used as a selection criterion for resistance to H. contortus infections in Merino sheep. PMID- 12088075 TI - Cytomegalovirus infection in a pig in South Africa. AB - An 8-week-old piglet with dyspnoea, bilateral mucopurulent nasal discharge and mouth breathing was euthanased and a necropsy was performed. Apart from histological evidence of diffuse rhinitis, large intranuclear inclusion bodies, pathognomonic forporcine cytomegalovirus infection, were detected within mucous glands on the nasal turbinates. This is the first such case to be diagnosed in South Africa. PMID- 12088076 TI - Clinical and serological response of wild dogs (Lycaon pictus) to vaccination against canine distemper, canine parvovirus infection and rabies. AB - Wild dogs Lycaon pictuis (n = 8) were vaccinated 4 times against canine distemper (n = 8) (initially with inactivated and subsequently with live attenuated strains of canine distemper) and canine parvovirus infection (n = 8) over a period of 360 days. Four of the wild dogs were also vaccinated 3 times against rabies using a live oral vaccine and 4 with an inactivated parenteral vaccine. Commercially available canine distemper, canine parvovirus and parenteral rabies vaccines, intended for use in domestic dogs, were used. None of the vaccinated dogs showed any untoward clinical signs. The inactivated canine distemper vaccine did not result in seroconversion whereas the attenuated live vaccine resulted in seroconversion in all wild dogs. Presumably protective concentrations of antibodies to canine distemper virus were present in all wild dogs for at least 451 days. Canine parvovirus haemagglutination inhibition titres were present in all wild dogs prior to the administration of vaccine and protective concentrations persisted for at least 451 days. Vaccination against parvovirus infection resulted in a temporary increase in canine parvovirus haemagglutination inhibition titres in most dogs. Administration of both inactivated parenteral and live oral rabies vaccine initially resulted in seroconversion in 7 of 8 dogs. These titres, however, dropped to very low concentrations within 100 days. Booster administrations resulted in increased antibody concentrations in all dogs. It was concluded that the vaccines were safe to use in healthy subadult wild dogs and that a vaccination protocol in free-ranging wild dogs should at least incorporate booster vaccinations against rabies 3-6 months after the first inoculation. PMID- 12088077 TI - Hydrogen proton magnetic resonance spectroscopy in autism: preliminary evidence of elevated choline/creatine ratio. AB - Hydrogen proton magnetic resonance spectroscopy is only beginning to be studied in autistic individuals. We report an association between hydrogen proton magnetic resonance spectroscopy choline/creatine ratios and severity of autism as measured by the Children's Autistic Rating Scale (Pearson r = .657, P = .04) in 10 autistic children. Hydrogen proton magnetic resonance spectroscopy choline/creatine ratio measures the concentration of cytosolic choline including free choline used in the synthesis of acetylcholine. Elevation in this ratio has been interpreted as a result of membrane degradation such as caused by a tumor or, alternatively, as a result of choline synthesis associated with increased cellular proliferation. Recent neuropathologic evidence has implicated disruption of acetylcholine transmission in the brains of autistic adults. A case-controlled study of hydrogen proton magnetic resonance spectroscopy choline/creatine ratios is warranted. PMID- 12088078 TI - Menkes gene study in the Chinese population. AB - We sought to determine the nature of the molecular defect causing Menkes' syndrome in the Chinese population. The DNA of four patients with typical Menkes manifestation was sequenced. Two pathologic genetic defects were identified; one of them is a nonsense mutation, whereas the other is a frameshift mutation. Both of these mutations are de novo. PMID- 12088079 TI - Alpha[11C] methyl-L-typtophan positron emission tomography in patients with alternating hemiplegia of childhood. AB - Based on previous reports suggesting a role of the neurotransmitter serotonin in the pathomechanism of alternating hemiplegia of childhood and speculation that it may be a migraine variant, we measured brain serotonin synthesis in children with alternating hemiplegia of childhood. Clinical and neurodevelopmental data, as well as standard uptake values in 25 brain regions and whole-brain serotonin synthesis capacity (unidirectional uptake rate constant or K-complex), were assessed in six patients with alternating hemiplegia of childhood (three girls and three boys; mean age = 7 6/12 years) using alpha[11C]methyl-L-tryptophan positron emission tomography (PET). The PET studies were performed interictally in three patients, during the ictal state in two patients, and postictally in one patient. The PET data were compared to those obtained interictally from six age matched patients with focal epilepsy (two girls and four boys; mean age = 7 8/12 years) and six non-age-matched apparently normal siblings of autistic children (two girls and four boys; mean age = 9 11/12 years). Patients with alternating hemiplegia of childhood studied in the ictal or postictal state showed increased serotonin synthesis capacity in the frontoparietal cortex, lateral and medial temporal structures, striatum, and thalamus when compared to controls, and subjects with alternating hemiplegia of childhood studied interictally. The involvement of these brain regions was consistent with the semiology of the hemiplegic attacks. In patients with interictal studies and in the controls, the PET scans revealed similar and bilaterally symmetric regional patterns of serotonin synthesis capacity. Increased whole-brain serotonin synthesis capacity (reported in migraine subjects without aura) was not found in the alternating hemiplegia of childhood group. There was no correlation between the neurodevelopmental scores and regional standard uptake values; however, patients with a larger estimated lifetime attack number showed greater delay in communication (P = .005) and daily living skills (P = .042). These studies suggest increased regional serotonergic activity associated with attacks in alternating hemiplegia of childhood. Furthermore, the attack number may have an effect on neurodevelopmental delay, thus supporting the notion that alternating hemiplegia of childhood may be a progressive disorder. PMID- 12088080 TI - Cerebrospinal fluid neuron-specific enolase following seizures in children: role of etiology. AB - Neuron-specific enolase, a marker for neuronal injury, is elevated following seizures in adults, but relatively few data exist on postictal neuron-specific enolase levels in children. This study measured cerebrospinal fluid (CSF) neuron specific enolase levels after seizures in 49 consecutive pediatric patients and investigated the role of seizure type, duration, and etiology in influencing neuron-specific enolase. Overall, there was no significant difference in neuron specific enolase levels between patients with seizures and a control group. However, 4 of the 49 seizure patients (8%) had neuron-specific enolase levels clearly above the normal range. Seizure patients with symptomatic etiologies had significantly increased neuron-specific enolase compared to cryptogenic/idiopathic or febrile seizures. The four individual patients with elevated cerebrospinal fluid neuron-specific enolase all had identified metabolic or genetic etiologies and presented with medically refractory status epilepticus. No individuals with cryptogenic/idiopathic or febrile seizures had abnormal neuron-specific enolase. There was no significant effect of seizure duration or type on cerebrospinal fluid neuron-specific enolase. In contrast to adults, acute seizure-induced neuronal injury in children as detected by neuron-specific enolase is rare and may occur primarily with severe symptomatic etiologies. Children with cryptogenic, idiopathic, or febrile seizures, including status epilepticus, are at relatively low risk for neuronal damage following seizures. PMID- 12088081 TI - Insulin resistance in epileptic girls who gain weight after therapy with valproic acid. AB - Valproic acid is effective for treatment of many types of epilepsy, but its use in epileptic patients can be associated with an increase in body weight that could interfere with treatment compliance. The weight gain may result from different mechanisms, but the exact pathogenesis is still unknown. To evaluate insulin sensitivity in adolescents who gained weight during treatment with valproic acid, we studied 20 girls with different types of epilepsy: 15 patients had primary generalized seizures, including absence seizures (3 cases), and 5 patients had partial seizures. After 1 year of valproic acid treatment, the obese patients had serum insulin levels significantly higher than patients who did not gain weight (51.4 +/- 25.3 versus 28.2 +/- 12.9). Moreover, we observed that epileptic patients who gained weight were also insulin resistant in comparison with nonobese epileptic subjects. At the end of treatment, all patients showed normal levels of serum testosterone, androstenedione, dehydroepiandrosterone sulfate, follicle-stimulating hormone (FSH), and luteinizing hormone. We found no significant correlation between insulinemia and serum valproic acid concentrations in obese and nonobese patients treated with valproic acid. Our study demonstrates that basal hyperinsulinemia and insulin resistance can be present in patients who develop obesity during valproic acid treatment. Therefore, these obese patients could be exposed to the risks related to these metabolic abnormalities; if these data are confirmed in longer studies, these side effects may raise some concerns about the safety of valproic acid. PMID- 12088082 TI - Prevalence estimation of Williams syndrome. AB - There are limited population-based data on the occurrence of Williams syndrome. We estimated its prevalence combining data from two investigations. One was an epidemiologic study originally designed to assess the prevalence and etiology of mental retardation among 30,037 Norwegian children born between 1980 and 1985 and living in Akershus County on January 1, 1993. The other investigation was a national survey of Williams syndrome. In the first study, 213 children were referred for evaluation, whereas the second study comprised 57 cases with Williams syndrome born between 1970 and 1992, who were referred for evaluation from all Norwegian counties. The epidemiologic study revealed three children with Williams syndrome, whereas one additional case complying with our demographic criteria was identified in the national survey, thus giving a prevalence of 1 in 7500. In all cases, a typical chromosome 7q11.23 deletion was detected. We also conclude that Williams syndrome is not an uncommon cause of mental retardation, with a prevalence of approximately 6% of patients with genetic etiology. PMID- 12088083 TI - Parameters for predicting favorable responses to botulinum toxin in children with cerebral palsy. AB - We sought markers for predicting a favorable outcome of botulinum toxin A injected to the lower-extremity muscles of 26 children with hemiplegic or diplegic cerebral palsy. Clinical assessment preceding and 1 month following injection included gross motor function measure, a modified Ashworth scale, and evaluation of range of motion of knee extension and ankle dorsiflexion. Response to treatment was classified based on a parent questionnaire. The 19 children (73%) considered by their parents as being good responders were compared to the 7 (27%) considered as being poor responders. In the good responders, the preinjection Ashworth scale (spasticity) was significantly higher (P < .05) and gross motor function measure scores (function) were lower (P < .05). Sixty-eight percent of the good responders were nonindependent walkers compared to 14% of the poor responders (P < .05). There were no differences in age, type of cerebral palsy, and dose of injection. An Ashworth scale indicating increased muscle tone, lower gross motor function measure scores, and nonindependent ambulatory status were predictive for a favorable response to botulinum toxin A injections and can guide patient selection and expectations of treatment outcome. PMID- 12088085 TI - Giant congenital melanocytic nevi in a patient with brain structural malformations and multiple lipomatosis. AB - We present a 9-year-old boy diagnosed from birth with giant congenital melanocytic nevi. He had central structural brain malformations of hemimegalencephaly of the right frontotemporal lobe and left occipitoparietal lobe, choroid plexus hypertrophy, and a Dandy-Walker variant. In addition, he developed multiple lipomatoses. These lesions were cutaneous except for two at the cerebellopontine angles, which were present from birth. This patient represents a rarely documented example of two histopathologies resulting in serious complications. The diagnostic issues and histopathologic process are discussed. PMID- 12088084 TI - Long-term tolerability and efficacy of lamotrigine in pediatric patients with epilepsy. AB - Accumulating data suggest that the antiepilepsy drug lamotrigine, which has been available for adult use for more than a decade, also confers broad-spectrum, well tolerated control of epilepsy in children. The current study--the open-label continuation phase of several short-term clinical trials--was conducted to assess the long-term tolerability and efficacy of lamotrigine as open-label adjunctive therapy or monotherapy in pediatric patients for a variety of seizure types and syndromes including partial seizures, absence seizures, and Lennox-Gastaut syndrome. Clinic visits occurred every 24 weeks throughout the treatment period. A total of 252 patients under 16 years of age were enrolled in the study. The numbers of patients exposed to at least 48 weeks, 96 weeks, and 144 weeks of treatment with lamotrigine were 185 (73.4%), 119 (47.2%), and 60 (23.8%), respectively, for an average duration of exposure of 96.7 weeks. The most common adverse events considered by the investigator to be drug related were dizziness (9.1%), somnolence (7.9%), nausea (6.3%), vomiting (5.2%), and headache (5.2%). The most common serious adverse events (regardless of suspected cause) included pneumonia (3.0%) and infection (1.9%). Investigators judged that the overall clinical status of three-fourths of the patients had improved at treatment weeks 48 and 96 relative to prelamotrigine clinical status. Lamotrigine administered as monotherapy or adjunctive therapy for an average of 2 years (96.7 weeks) was well tolerated and effective in pediatric patients with partial or generalized epilepsy. These results complement and extend the large body of data demonstrating the tolerability and efficacy of lamotrigine with short- and long term use in adults. PMID- 12088087 TI - Ictal electroencephalography in a case of benign centrotemporal epilepsy. AB - The purpose of this research was to publish a complete ictal electroencephalographic (EEG) record of a child with benign centrotemporal epilepsy. The record includes a brief description of the patient's epilepsy syndrome, analysis of interictal and ictal EEG records, and a description of corresponding clinical ictal events. Ictal EEG activity appeared over the right rolandic cortex and spread to adjacent sites. Initial rhythmic activity was followed by bilateral series of sharply contoured slow waves. The electric field of the seizure discharge changed during the seizure. Tangentially oriented dipolar field was found in the middle part of the seizure. Evolution of clinical seizure phenomena roughly corresponded to propagation of the ictal discharge. In terms of topography and electromorphology, this ictal record was similar but not completely identical with those reported previously. Collection of further ictal records in benign rolandic or centrotemporal epilepsy of childhood and other epilepsy syndromes is necessary. PMID- 12088086 TI - Interhemispheric reorganization of motor hand function to the primary motor cortex predicted with functional magnetic resonance imaging and transcranial magnetic stimulation. AB - The objective of this study was presurgical assessment of reorganization of motor hand function in an 11-year-old girl with intractable epilepsy and a right-sided hemiplegia resulting from an extensive perinatal left hemispheric stroke. Prior to a left functional hemispherectomy, functional magnetic resonance imaging (MRI) showed that both nonparetic and paretic motor hand function predominantly activated the right primary motor cortex, whereas no activation was found in the left hemisphere. Transcranial magnetic stimulation of the right central area yielded responses in both the nonparetic and the paretic hand, whereas no responses were obtained after stimulation of the affected hemisphere. Both techniques indicated that motor function was mediated by corticospinal fibers originating from the undamaged (primary) motor cortex and predicted no further loss of motor hand function after surgery. Indeed, subsequent functional hemispherectomy induced no new sensorimotor deficits. Functional MRI was repeated 22 months after surgery and matched preoperative sensorimotor functional MRI findings, confirming reorganization of the primary motor cortex. No additional reorganization was introduced by surgery. PMID- 12088088 TI - Periventricular nodular heterotopia: report of a pediatric series. AB - Periventricular nodular heterotopia is a malformation that occurs in both males and females and is associated with a variety of clinical and neuroradiologic signs. A gene called filamin-1 (FLN-1) has recently been identified. We review the clinical and imaging findings from a series of pediatric patients with periventricular nodular heterotopia. Five patients (three males and two females; age range = 4-18 years) were investigated. In our series, periventricular nodular heterotopia can be the common denominator in different conditions. Periventricular nodular heterotopia can occur alone or be associated with cortical malformations. Epilepsy was present in three of the five patients and was resistant to drugs in one female. Mental retardation was present in three of the five patients. Two male patients had normal intelligence, with no cortical anomalies; patient 3 had unilateral periventricular nodular heterotopia. The associated malformations were more severe in the female patients and slight only in patient 1. The two females showed anomalies rarely reported in association with bilateral periventricular nodular heterotopia. We believe that other genes can be involved in children with atypical neuroradiologic periventricular nodular heterotopia. No mutations were detected in 6 of the 48 exons of the FLN-1 gene, although this does not allow any definitive conclusions to be reached. We conclude that our series of patients with periventricular nodular heterotopia clearly highlights the complexity of the clinical, neurologic, and neuroradiologic characteristics associated with this malformation. PMID- 12088089 TI - Combined partial deficiencies of carnitine palmitoyltransferase II and mitochondrial complex I presenting as increased serum creatine kinase level. AB - Increased serum creatine kinase level is a marker of neuromuscular disorders. When combined with exercise intolerance, muscle cramps, fatigue, myoglobinuria, or muscle weakness, metabolic myopathies of a variety of causes should be considered. We encountered an adolescent male with a persistently high serum creatine kinase level and chronic fatigue who was found to have combined partial defects of carnitine palmitoyltransferase II and mitochondrial complex I. Metabolic myopathy may present with chronic fatigue and a persistently high serum creatine kinase level but without muscle weakness and may be attributable to combined enzyme defects. PMID- 12088090 TI - Unilateral auditory neuropathy: case study. AB - This article reports on an 11-year-old boy who was diagnosed with unilateral auditory neuropathy. After failing his annual medical and school hearing screenings, he was referred for audiologic testing, which identified a profound sensorineural hearing loss in his left ear that has remained stable for the past 3 1/2 years. Subsequently, otoacoustic emissions revealed findings that were consistent with an auditory neuropathy. Neurologic evaluation was remarkable only for a left ear hearing loss. Radiologic studies were unremarkable. Auditory neuropathy is a condition in which patients, on audiologic evaluation, are found to have normal outer hair cell function and abnormal neural function at the level of the eighth nerve. These patients, on clinical testing, are found to have normal otoacoustic emissions, whereas auditory brainstem response audiometry reveals the absence of neural synchrony. Unlike space-occupying lesions, radiologic evaluation reveals normal results. This case is noteworthy because only a handful of the cases of auditory neuropathy reported to date have been unilateral. PMID- 12088091 TI - Children with irreversible brain damage associated with hypothyroidism and multiple intracranial calcifications. AB - Children who develop clinical hypothyroidism in early childhood have various degrees of irreversible brain damage, albeit less severe than cases detected by neonatal screening test for hypothyroidism in the first months of the life. We report three patients with hypothyroidism of childhood onset after a normal neonatal thyroid-stimulating hormone screening who showed deceleration in linear growth, spasticity in the lower limbs with deformity, mild intellectual impairment, and multiple calcifications in the basal ganglia and subcortical areas. The neurologic symptoms were not progressive but were irreversible in spite of thyroxine treatment. Motor disturbances commonly observed in postnatal onset hypothyroidism are similar to those of cerebral palsy. Specific distribution of intracranial calcifications may result from metabolic derangement as a result of hypothyroidism, although the mechanism of calcification is not fully understood. We emphasize the need to re-evaluate thyroid function in diplegic patients with specific intracranial calcifications but normal neonatal thyroid-stimulating hormone screening. PMID- 12088092 TI - Trends in cabin air quality of commercial aircraft: industry and passenger perspectives. AB - The small air space available per person in a fully occupied aircraft passenger cabin accentuates the human bioeffluent factor in the maintenance of air quality. The accumulation of carbon dioxide and other contributions to poor air quality that can occur with inadequate ventilation, even under normal circumstances, is related to the volume of available air space per person and various ventilation rates. This information is compared with established air quality guidelines to make specific recommendations with reference to aircraft passenger cabins under both normal and abnormal operating conditions. The effects of respiration on the air quality of any enclosed space from the respiration of a resting adult are estimated using standard equations. Results are given for different volumes of space per person, for zero air exchange, and for various air change rates. The required ventilation rates estimated in this way compared closely with results calculated using a standard empirical formula. The results confirm that the outside air ventilation required to achieve a target carbon dioxide concentration in the air of an occupied enclosed space remains the same regardless of the volume of that space. The outside air ventilation capability of older and more recent aircraft is then reviewed and compared with the actual measurements of cabin air quality for these periods. The correlation between calculated and measured aircraft cabin carbon dioxide concentrations from other studies was very good. Respiratory benefits and costs of returning to the 30% higher outside air ventilation rates and 8% higher cabin pressures of the 1960s and 1970s are outlined. Consideration is given to the occasional occurrence of certain types of aircraft malfunction that can introduce more serious contaminants to the aircraft cabin. Recommendations and suggestions for aircraft builders and operators are made that will help improve aircraft cabin air quality and the partial pressure of oxygen that is available to passengers at minimal cost. Also suggested are some measures that passengers can take to help improve their comfort and decrease their risk of illness, particularly on long-haul flights. PMID- 12088093 TI - Age-environment and gene-environment interactions in the pathogenesis of Parkinson's disease. AB - Parkinson's disease (PD) is a common neurodegenerative disease characterized by dopaminergic cell death and deposition of Lewy bodies within the substantia nigra of the midbrain. Although the major risk factors for PD are aging and environmental factors, there is an important genetic component. An age-related change in xenobiotic metabolism alters the metabolism of and net exposure to, environmental neurotoxins. Genetic variability in xenobiotic metabolism may similarly increase the susceptibility to PD by altering the metabolism of neurotoxins. Genetic studies of rare familial cases of PD indicate a central mechanistic role for the aggregation of alpha-synuclein, a protein found in Lewy bodies. Environmental factors like pesticides and heavy metals can also influence alpha-synuclein aggregation. Common final pathways for aging, environmental, and genetic mechanisms can thus exist, involving both direct neurotoxicity and alpha synuclein aggregation. PMID- 12088094 TI - Somatotype and disease prevalence in adults. AB - We examined the association between the somatotype and its main components (endomorphy, mesomorphy and ectomorphy), and the prevalence of several chronic diseases. The data were obtained from a cross-sectional survey designed to assess somatotype and morbidity with special reference to most often diagnosed diseases. The study population comprised 524 men and 250 women. The subjects underwent laboratory tests and clinical and anthropometric examinations. Of all examined workers, 94.8% fell into the five somatotype categories; of these, 394 were endomorphic mesomorphs. The most common somatotype was endomorphic mesomorph for men and mesomorph-endomorph for women. In five disease groups, prevalence was significantly related to a somatotype. Mesomorphic endomorphs most frequently suffered from digestive system diseases (40.6%, p < 0.05), neuroses (30.1%, p < 0.05), and radiculitis lumbosacralis (15.4%). The prevalence of arterial hypertension in mesomorph-endomorphs (37.1%), endomorphic mesomorphs (35.5%), and mesomorphic endomorphs (34.3%) was equal. In both genders, those with the highest endomorphy and mesomorphy and the lowest ectomorphy, grouped by cluster analysis, were those who suffered most frequently from arterial hypertension and liver disease. The authors conclude that the somatotype having a dominant mesomorphy and marked endomorphy constitutes a risk factor as a particular predisposition toward certain diseases and requires body weight control. PMID- 12088095 TI - What's in a condom?---HIV and sexual politics in Japan. AB - Utilizing a range of ethnographic data from an AIDS hotline, a women's shelter, a night club, AIDS campaigns, news articles, and interviews with health bureaucrats, this paper explores the history of AIDS in Japan and the ways in which official practices reproduce systems of domination. This paper examines the official categories of "foreign woman" and "prostitution" as discursive strategies of containment, and argues that nationalist discourses and representations of sexuality are closely linked in maintaining relations of power. PMID- 12088096 TI - The tension between overt talk and covert emotions in illness narratives: transition from clinician to researcher. AB - In this article I discuss my interview experiences as joint constructions with Finnish parents whose children had been diagnosed with cancer. In doing so both the interviewer and interviewees became engaged as anxious "defended subjects" in an intersubjectivity of acknowledging but unable to talk of "how it really is" to have a child diagnosed with cancer. This appeared as tensions between overt talk and covert emotions. In the overt talk the parents underlined the manageable aspects of the illness. The speechless and painful part of the illness experience appeared as covert emotions that the interviewer was able to recognise and contain. Only when both the overt talk and the covert emotions were taken into account did it open up a more comprehensive way to understand the depth of the illness suffering. This is illustrated with one interview as an example. PMID- 12088097 TI - Gender, violence and HIV: women's survival in the streets. AB - In this article I propose that gender inequality promotes--directly or indirectly -vulnerability to HIV as a consequence of a multidimensional violence (structural, symbolic and physical) experienced by injection drug using (IDU) women in The Mission District (San Francisco). Given the female subordinated position stipulated by the street ideology, I analyze how drug dependence afforded by precarious strategies of subsistence places IDU women under multiple dangers and threats. In this setting, unequal gender relations are part of a complex system of transactions in the street economy and a way to reduce or increase the everyday violence. Facing multiple dangers and risks, some women adopt a subordinated position, some try to negotiate the conditions of the exchanges and the others resist the exploitation. Finally, everyday violence under conditions of gender inequality and scarcity of resources imposes a logic defined by the challenge of survival under the threat of immediate dangers, which transform HIV into a secondary risk. PMID- 12088098 TI - HIV, disease plague, demoralization and "burnout": resident experience of the medical profession in Nairobi, Kenya. AB - This paper describes the experiences of physicians-in-training at a public hospital in Nairobi, Kenya, where medical professionals practice in an environment characterized by both significant lack of resources and patients with HIV/AIDS in historically unprecedented numbers. The data reported here are part of a larger study examining ethical dilemmas in medical education and practice among physicians in East Africa. A questionnaire and semi-structured interview were completed by fifty residents in four medical specialties, examining social and emotional supports, personal and professional sources of stress, emotional numbing and disengagement from patients and peers, and symptoms of post-traumatic stress and depression. The factors affecting resident well-being are found in this study to be more complex than previous interviews suggested. This study highlights the fact that as a result of working in an environment characterized by poor communication among hospital staff as well as a lack of resources and high numbers of patients with HIV/AIDS, residents' perceptions of themselves- their technical proficiency, their ability to care and feel for others and themselves, and for some their entire sense of self--are significantly affected. Also affected are the patients they work to treat. PMID- 12088099 TI - Agonias: the social and sacred suffering of Azorean immigrants. AB - Agonias, meaning "the agonies," is a culture-specific somatic phenomenon experienced by Azorean immigrants. Although the community's health providers conceptualize agonias as an "anxiety disorder," interviews with community members revealed a more complex phenomenon. For them, agonias is a somatomoral experience -where the somatic, the social, the religious and the moral are inextricably linked. Because agonias connects things that, from the traditional medical perspective, should not be connected, such as mind, body, spirit, and community, it defies our psychiatric categorisation and goes beyond disciplinary boundaries. Agonias is a dynamic multivocal symbol that is not just an inanimate signifier but also a therapeutic act. On an individual level, it connects the sufferer with others and with God, transforming the interpersonal and divine space. On the societal level, it connects a community, losing its way of life, to the past and to its identity, preserving its social and religious traditions. PMID- 12088100 TI - Changes in apoptosis, mitosis, Her-2, p53 and Bcl2 expression in breast carcinomas after short-term tamoxifen treatment. AB - The short-term (7 days) effect of tamoxifen on apoptosis and mitosis index, p53, Bcl2 and Her-2/neu/c-erb2 expression in invasive ductal mammary carcinoma was studied histologically in the diagnostic biopsy and surgically removed tumor tissue of 10 patients. Following tamoxifen treatment expression of HER-2 and p53 decreased but Bcl2 remained unchanged. Mitotic activity decreased slightly, but not significantly. Apoptotic activity increased in six cases in the second sample compared to the values measured of the first biopsy. These changes may be attributed to the effect of antiestrogen therapy. PMID- 12088101 TI - MUC1 expression in human breast cancer cells is altered by the factors affecting cell proliferation. AB - Increased expression of the epithelial mucin MUC1 has been linked to tumor aggressiveness in human breast carcinoma. In the present study, we have investigated if the factors affecting cells proliferation could influence MUC1 mucin biosynthesis and shedding from cell surface into the culture medium in two human breast cancer cell lines: MCF-7 (ER+) and MDA-MB-231 (ER-). Using MCF-7 line we found that estradiol at a concentration of 10(-7) M increased [3H]glucosamine incorporation into mucin in cell lysate approximately twofold in comparison with control cultures, and a similar increase was observed in the culture medium. The selective estrogen receptor modulator, tamoxifen (at concentrations of 10(-6) M and 10(-5) M) had a little inhibitory effect. MDA-MB 231 cells in culture were stimulated with phorbol ester PMA, the protein kinase C activator. We noted that PMA greatly stimulated MUC1 synthesis and its shedding to culture medium and that this effect was abolished by protein kinase C specific inhibitor--bisindolylmaleimide. PMID- 12088102 TI - Expression of c-erbB-2 in node negative breast cancer does not correlate with estrogen receptor status, predictors of hormone responsiveness, or PCNA expression. AB - The aim of this study was to analyse the relationships between the expression of c-erbB-2, estrogen receptor (ER), progesterone receptor (PR), Bcl-2 and PCNA in node negative breast cancer. Expression of these markers was determined by HercepTest, by immunohistochemistry and quantified by morphometry in the group of 125 selected breast carcinoma patients with broad spectrum of histological types and grades. Multivariate statistical analysis revealed only relationships between ER/PR, ER/Bcl-2, ER/grade and ER/age. There was not found any significant relationship between c-erbB-2 expression and any other immunohistochemical marker, apocrine metaplasia, histological type or patient characteristics. The same result was found in complete group of tumors as well as in individual groups divided according to histological type. These results indicate that in node negative breast tumors, c-erbB-2 expression does not correlate inversely with hormone receptor status and hormone responsiveness like previously reported metastasising breast cancer and that the prognostic significance of c-erbB-2 expression in these tumors is not clear. PMID- 12088103 TI - Possible genotoxic activity of extracts of Bryonia alba roots on human lymphocytes and transformed cells. AB - Bryonia alba roots (BAR) are widely used as an adaptogenic and restorative drug with immunomodulatory and stress-protective properties that increase the non specific resistance of an organism toward harmful stimuli. Potential genotoxic activity of aqueous and methanol extracts of BAR was studied on human normal (lymphocytes) and transformed (HeLa and Caco-2) cells using single cell gel electrophoresis (the comet assay). The results obtained did not show any evidence of genotoxic effects of BAR. PMID- 12088105 TI - Clinical significance of 5-S-cysteinyldopa monitoring in patients with malignant melanoma. AB - 5-S-cysteinyldopa is a precursor of melanin. Its serum and urinary level can reflect melanoma progression. In this study the concentration changes of 5-S-CD in melanomas of all stages were examined, and patients were monitored during and after treatment. Serum samples were taken from 479 melanoma patients with different Stages on 1924 occasions, from June 1996 to December 2000. Levels of 5 S-CD were determined by HPLC. The mean/median value of 5-S-CD in the Stage I-II III patients and in the control group ranged around the normal level. Significant difference was found between Stage III and Stage IV as well as between patients with no evidence of disease and patients with tumor burden. In Stage IV 69.7% sensitivity, 61.5% specificity and 79.3% positive predictive value were evaluated. The survival of patients with normal 5-S-CD level (n=235) differed significantly from cases having elevated marker concentration (n=244). One hundred eighty cases were regularly monitored on 1210 occasions. Recurrence development was noticed in 57 patients. In 24.6% of these patients suffering from any type of disease progression the increasing marker level preceded the detection of metastasis by conventional methods. Serum 5-S-CD in Stage IV is sensitive enough to detect distant metastasis, and its predictive value has a great importance. It is a reliable marker for monitoring the clinical course in malignant melanoma. PMID- 12088104 TI - Concomitant radiotherapy and metronomic temozolomide in pediatric high-risk brain tumors. AB - Temozolomide, an oral alkylating agent has a significant activity in preclinical testing and in clinical trials in adults and children as well. Penetration across the blood brain barrier has been documented. In adult and pediatric phase I and II trials a five-day every 28 days schedule was first approved for clinical use. With respect to temozolomide proximal mechanism of resistance, and further to increase dose intensity, new schedules are proposed to use more prolonged drug exposure. Higher doses of metronomic temozolomide were piloted. Eight children with poor prognosis brain tumors were eligible. Treatment consisted from concomitant radiotherapy given 1x170 cGy, 5d/wk, for total dose 55/56 Gy, together with temozolomide 90 mg/m2/day for 42 days. No further dose escalation has been planned for this group of patients. Myelosuppression was the primary toxicity, occurring around day 21. Nonhematologic toxicities were infrequent and in no case dose limiting toxicity (DLT) occurred. The most common nonhematologic toxicity was vomiting, effectively managed with antiemetics. Six responses were documented. The best responses (CR) were seen in 2 patients with high-risk medulloblastomas, who have progressed early after neurosurgery. Furthermore, one more patient had CR and 3 patients PR at the end of temozolomide treatment. We have piloted novel dose schedule of temozolomide and evaluated clinical toxicities in a cohort of 8 children. This is the first study to report feasibility and tolerability of 90 mg/m2/day of temozolomide treatment in metronomic fashion. In addition, we have documented encouraging responses in children with medulloblastomas, progressing early after their initial surgery. PMID- 12088106 TI - Freeze dried ondansetron: first observations with the fast dissolving oral antiemetic Zofran Zydis for the prophylaxis of the cisplatin-induced emesis in gynecological cancer patients. AB - Oral antiemetics are seldom taken by patients (women, children or those having tumors of the buccal area, mouth or esophagus), who find it difficult to swallow. In addition to anatomical reasons, the side effects of cytostatics require medication. Fast dissolving ondansetron is a new preparation, which instantaneously disintegrates and disperses in the saliva. The purpose of this paper was to evaluate the antiemetic effectivity of the product Zofran Zydis. Thirty six patients treated with cisplatin (50 mg/m2) in 55 chemotherapy courses were given 2x8 mg fast dissolving ondansetron in a prospective non randomised study. 75% complete response and 11% major response rates were found. Authors conclude that fast dissolving ondansetron is a new and effective preparation that enriches the panel of available supporting drugs. PMID- 12088107 TI - The cost effectiveness of dual phase 201Tl thyroid scan in detecting thyroid cancer for evaluating thyroid nodules with equivocal fine-needle aspiration results: the preliminary Taiwanese experience. AB - The purposes of this study were to assess the helpfulness of dual phase 201Tl thyroid scan for differentiating malignant from benign thyroid lesions in cases of thyroid nodules with equivocal fine-needle aspiration (FNA) biopsy results. In addition, for thyroid nodules with equivocal FNA biopsy results, we try to make a decision analysis model compared the FNA biopsy alone strategy (strategy A) with decision strategy for the assistance of dual phase 201Tl thyroid scan (strategy B) before diagnostic thyroidectomy as thyroid cancer evaluation strategies for hypothetical cohorts of estimated 17,280-29,160 Taiwanese patients/per year with equivocal FNA biopsy results. Based on the findings of surgical histopathology, dual phase 201Tl thyroid scan sensitivity, specificity, and accuracy were 100%, 90%, and 96%, respectively, in cases of 27 thyroid nodules with equivocal FNA biopsy results. In cost effectiveness analysis, the strategy B showed a cost saving of 16,340,480-27,574,560 US dollars in unnecessary diagnostic thyroidectomy cost. The total cost of strategy B showed a cost saving of 13,932,232-23,520,564 US dollars than that of strategy A. The preliminary data indicate that dual phase 201Tl thyroid scan can save the cost of unnecessary diagnostic thyroidectomy in Taiwanese patients with equivocal FNA biopsy results. In addition, we may provide a noninvasive diagnostic method--dual phase 201Tl thyroid scan, as the first priority for Taiwanese patients with equivocal FNA biopsy before diagnostic thyroidectomy under the coverage of the national health insurance system in Taiwan. PMID- 12088108 TI - Prostate cancer and the food supplement, PC-SPES. Minireview. AB - Cancer of the prostate has now become the most frequently diagnosed cancer in the male population in America and indeed its incidence has increased in many westernised countries. Surgery, radiotherapy and androgen ablation form the mainstay of its treatment. However, these treatment modalities are ineffective in a large percentage of patients due to grade of tumor at presentation, sensitivity to radiotherapy, hormone-insensitive profile of tumor cell population, etc. Because of the woeful ineffectiveness of these standard treatment modalities on the overall survival rate from prostate cancer in the past fifty years, a more radical approach to the treatment of this cancer is justified. To this end some of the latest innovative approaches are now being tested as tools in prostate cancer research, including those pioneered in molecular and cellular biology and immunology. There has also arisen a considerable interest in the modification of diet and the use of herbs and a great interest has been stimulated in PC-SPES, a herbal food supplement sold in the USA. This product, which is composed of eight herbs, is recommended as a food supplement for those suffering from prostate cancer and there are many anecdotal, scientific and clinical claims for its efficacy. This is a review of some of the in vivo and in vitro research carried out on this product. PMID- 12088109 TI - A case-control study of lung cancer in Polish women. AB - This study was undertaken to examine the influence of active and passive smoking, cancer family history, occupational exposure, usual diet and alcohol consumption on female lung cancer risk. A total of 242 women with histologically confirmed primary lung cancer and 352 healthy controls were involved in the study. All subjects were interviewed in the hospital. Data were analysed using multivariate logistic regression. Multivariate analysis has shown that smoking was the most strongly active risk factor in female lung cancer. Positive dose-response relationship was observed between lung cancer risk and number of pack-years. Passive smoking exposure during childhood significantly increased the risk (OR=2.65). There was also observed a significantly increased risk of lung cancer among women who had siblings with history of cancer (OR=3.42). Occupational exposure to coal dust, acid fumes (sulphuric and/or hydrochloric) and materials used for rubber making significantly increased the risk. Frequent intake of carrots (at least five times a week) and also daily intake of other vegetables significantly lowered the risk (OR=0.13, OR=0.24). A significant protective effect was also observed in women frequently using margarine on bread (OR=0.14). Vodka drinkers showed significantly higher risk than non-drinking women. The analysis of dose-response relationship in reference to vodka drinking also confirmed significant influence of this factor on the risk. PMID- 12088110 TI - Nonmelanoma skin cancers and risk of subsequent malignancies: a cancer registry based study in Bulgaria. AB - Analysis of new primary tumors following nonmelanoma skin cancers (NMSC) has a public health and risk assessment interest, as well as potential implications for etiologic inference. The aim of this study is to evaluate the risk of the second primary tumors occurrence after NMSC development. A cohort of 2620 cases (1335 males and 1285 females) with nonmelanoma skin cancers registered in Bulgarian National Cancer Registry in 1993 was examined. The follow-up period represented a total of 15,856 person-years at risk. Over this period 128 (83 in men, 45 in women) new secondary tumors were established. After NMSC appearance, both genders show higher risk for the second primary tumors occurrence. This risk is greater for men. For both sexes after NMSC development there is increased risk for occurrence of second primary cancers of head and neck, thyroid, lung, larynx, bladder, colon, as well as cutaneous malignant melanoma, non-Hodgkin's lymphoma and leukemias. The results of considering only basal cell carcinomas show an elevated risk in patients of both sexes for appearance of second primary tumors of head and neck, bladder, larynx, lung and colon as well as non-Hodgkin's lymphoma and cutaneous malignant melanoma. The results of considering only squamous cell carcinomas show an increased risk in persons of both sexes for development of second primary cancers of head and neck, skin, thyroid, lung, stomach, as well as leukemias, non-Hodgkin's lymphoma and cutaneous malignant melanoma. The association between NMSC and subsequent increased risk for appearance of second primary skin and visceral tumors determine the necessity of monitoring the patients with NMSC. PMID- 12088111 TI - Molecular diversity of gammadelta T cells in peripheral blood from patients with B-cell chronic lymphocytic leukaemia. AB - To characterize circulating gammadelta T cell subpopulations in B chronic lymphocytic leukaemia patients (n=30), TCR Vgamma and Vdelta gene-segment use was analyzed by RT-PCR using a panel of subfamily-specific oligonucleotide primers. All results were compared with those obtained with specimens from healthy donors (n=10). The cells expressing Vdelta1+ TCR displayed the highest relative increase in B-CLL patients (particularly observed in 60% of cases), but Vdelta3+ T lymphocytes also expanded in leukaemic peripheral blood (10% of studied cases). Both mentioned gammadelta T cell subsets were significantly more frequent in the most severe stages of disease--Rai III+IV. The analysis of Vgamma region usage in TCR formation revealed that gammadelta T cells from B-CLL patients predominantly expressed a Vgamma9 segment (26 of 30 cases), usually linked to Cgamma1 region. It should be noticed that the dominant TCR genes expression in a 50% of healthy donors was Vdelta2+/Vgamma9+, however, Vgamma4 and Vgamma8 transcripts were also observed (2 and 3 of 10 cases, respectively). The above results indirectly indicate that gammadelta T lymphocyte expansion was driven by the oligo- or polyclonal proliferation and can reflect specific response against the autologous tumor cells. PMID- 12088112 TI - Immune-mediated complications during interferon alpha therapy in chronic myelogenous leukemia. AB - Several prospective randomized studies have shown that the treatment of chronic myeloid leukemia with interferon alpha (IFNalpha) prolongs the survival by comparison with conventional chemotherapy. However, long-term treatment with Interferon alpha can produce or exacerbate immune-mediated complications (IMC). The purpose of this study was to analyze the experience with IMC in patients with chronic myelogenous leukemia (CML) undergoing IFNalpha treatment. The occurrence of IMC was evaluated in 76 patients (47 male; 29 female) with Philadelphia chromosome (Ph)-positive CML. Diagnostic criteria of IMC were performed in patients with symptoms suggestive of particular disorders. Well-documented and clinically evident complications developed in 7 patients after a median of 19 months (range 1-84) of IFNalpha treatment. These included 9.2% patients with Ph positive CML treated with IFNalpha-containing regimens. Hypothyroidism (H) occurred in 1 patient (1.3%), immune-mediated hemolysis (HEM) in 2 patients (2.6%) and connective tissue disorders (CTD) in 4 patients (5.3%) (2 systemic lupus erythematosus--SLE, 1 Raynaud's phenomena and 1 mixed connective tissue disease--MCTD). IFNalpha was discontinued in 3 patients and the dose was reduced in 2 patients. Five of 7 patients (75%) with immune-mediated complications had some degree of cytogenetic response at the time of the event. The association with female sex was strong and significant (86% vs 33.6%, x2; 48; p = 0.02). The frequency of IMC of clinical relevance with interferon alpha therapy in CML increased (long-term therapy). The patients treated with interferon alpha should be monitored for signs and symptoms of autoimmunity. PMID- 12088113 TI - Expression of cell-cycle related proteins in Helicobacter pylori gastritis and association with gastric carcinoma. AB - Helicobacter pylori (H. pylori) infection is associated with changes in epithelial turnover, through their significance of these in gastric carcinogenesis is still controversial. The purpose of this study was to determine the influence of H. pylori infection on cell proliferation and the relation with the cell-cycle regulators, and finally to provide insights into the mechanism by which H. pylori may lead to gastric carcinogenesis. We investigated Ki-67, p53, p21(Waf1/Cip1), cyclin D1 expression in 55 patients with H. pylori gastritis, and compared the results with patients those of non-H. pylori gastritis patients (n=21), gastric adenocarcinoma patients (n=8) and samples with normal gastric mucosa (n=12). Gastric biopsies were histologically evaluated for inflammatory reaction, intestinal metaplasia and atrophy according to the Sydney system. Overexpression of Ki-67, p53, p21(Waf1/Cip1) and cyclin D1 was found in H. pylori gastritis patients (32.7%, 10.9%, 20.0% and 7.3%, respectively), whereas only scattered expression in cells in the neck region of the crypts, but no overexpression was found in gastric antral epithelial cells in biopsy specimens from patients with non-H. pylori gastritis and noninflammed mucosa. A significant relationship was found between the grade of H. pylori colonization and Ki-67, p53, p21(Waf1/Cip1) and cyclin D1 expression. Expression was significantly higher in patients with intestinal metaplasia with atrophy, whereas no overexpression was found in patients without intestinal metaplasia with atrophy (p=0.05). H. pylori infection is associated with increased cell proliferation, increased epithelial DNA damage, and atrophy, which might contribute to the development of gastric cancer. Even if the exact mechanism has not been elucidated yet, our results suggest that H. pylori infection acts as a cofactor in gastric carcinogenesis. PMID- 12088115 TI - Breast cancer acute radiotherapy morbidity evaluated by different scoring systems. AB - Reporting of the outcome of radiotherapy is not satisfactory without a description of the treatment-related side effects. The purposes of this paper were: (1) to evaluate the frequency and the severity of collateral skin reactions in a group of breast cancer patients; (2) to report the acute reactions using some current scoring systems and to compare the application of them, and (3) to investigate the variation between intra- and interobservers using these different scales. We studied 108 breast cancer patients who, after surgical treatment, received adjuvant radiotherapy. Clinical skin evaluation was always performed by the same radiotherapist the last day of treatment, and the collateral radiation effects were photographed at that moment to facilitate later evaluations by another two expert doctors. Normal tissue damage was scored according to the Radiation Therapy Oncology Group/The European Organisation for Research, and Treatment of Cancer/ (RTOG/EORTC), the Danish, the European, and the Biomed2 side effect scales. The most frequent acute complications found were erythema (91.7%), dry desquamation (29.6%) and moist desquamation (35.2%). The reactions were classified as severe in 13.9, 23, 18.5 and 13% of the patients with each of the different systems used, respectively. The concordance between the scoring of radiation-induced side effects on the skin assessed by direct observation of the patients or by examination of the photographic document was sufficient. This is a warrant of accuracy in the evaluation of acute normal tissue lesions. Our results allow us to state the advantage of the RTOG system over the others in terms of evaluating the acute effects produced by radiotherapy of women with breast cancer. PMID- 12088114 TI - Homocamptothecin-daunorubicin association overcomes multidrug-resistance in breast cancer MCF7 cells. AB - The multidrug-resistance (MDR) status of a novel camptothecin analogue, homocamptothecin (hCPT), was investigated in human colon adenocarcinoma HT29 cells, myelogenous leukemia K562 cells and breast carcinoma MCF7 cells. The cytotoxicity of hCPT was not sensitive to the MDR status in K562 cell lines. However, its cytotoxicity was altered by MRP1, but not Pgp, in naturally MRP1 expressing HT29 cells, and etoposide- and doxorubicin-resistant MCF7/VP and MCF7/DOX cells, respectively. These cells were sensitized to hCPT in presence of MK571, probenecid but not verapamil. These results led to consider hCPT as a substrate for MRP1 and a potential modulator of MRP1 activity. The relationship between the cytotoxic effect of anthracyclines and their nuclear localization had been previously demonstrated. We show that MRPI mediated the daunorubicin (DNR) efflux in MCF7/VP and MCF7/DOX cells. The combination of sub-toxic doses of hCPT with DNR resulted in the potentiation of DNR activity, well-correlated with an increase in its nuclear accumulation in MCF7/VP cells. Simultaneous pattern was shown to provide higher cytotoxic response than sequential one. In agreement, hCPT increased also the DNR nuclear accumulation in low MRP1-expressing MCF7/DOX cells. However, the enhancement of cytotoxicity in the DNR-hCPT combination was poorly correlated with the nuclear concentration of DNR in MCF7/DOX cells. In addition to the increase in DNR accumulation, the potentiation of DNR activity by hCPT in MCF7/DOX cells implied a synergistic mechanism between both drugs. These data suggest that the present topoisomerase I/II inhibitors combination may be of clinical interest to overcome MDR phenotype in DNR-treated breast cancer patients. PMID- 12088116 TI - The use of molecular markers in farnesyltransferase inhibitor (FTI) therapy of breast cancer. AB - The hypothesis that pharmacologic inhibitors of Ras can be effective anti-cancer agents has led to the development of Farnesyltransferase inhibitors (FTIs). These agents inhibit the requisite processing of a number of cellular proteins including Ras. FTIs have shown good anti-tumor efficacy and little toxicity in preclinical models and based on these results, numerous clinical trials are currently underway to evaluate the clinical potential of these agents in patients with cancer. However, contrary to the ideas that led to their design, mechanistic studies have not confirmed that they inhibit tumors through the inhibition of Ras. FTIs inhibit the growth of a broad variety of human tumor cells in vitro and studies to date have not identified cellular characteristics that predict the antitumor efficacy of this class of agents. We have studied a panel of breast cancer cell lines that differ widely in their sensitivity to FTI in order to determine which molecular characteristics may determine sensitivity to this class of agents. In these cells we find that FTI sensitivity does not correlate with the relative expression of Ras isoforms or the inhibition of Ras processing, growth factor signaling, expression of estrogen receptor or the overexpression of growth factor receptors. Looking for other molecular correlates of FTI sensitivity we have compared the activity of farnesylprotein transferase (FPTase) among these cells and although we find no overall correlation with FTI sensitivity, we find that two cell lines with unusually low FPTase activity are sensitive. Comparing p53 genotype with FTI-sensitivity we find that although most cell lines in our panel have mutant p53, all three cell lines with wild-type p53 are quite sensitive to FTI. In fact, MCF-7 cells which have both wild-type p53 and the lowest FPTase activity are the most FTI-sensitive cell type we have ever seen. Although these studies do not identify any single molecular marker that can accurately predict FTI sensitivity in breast tumors, they highlight the potential roles of FPTase activity and p53 function for further analysis. PMID- 12088117 TI - The role of dietary long-chain n-3 fatty acids in anti-cancer immune defense and R3230AC mammary tumor growth in rats: influence of diet fat composition. AB - We determined if long-chain n-3 fatty acids fed as part of a: (1) high polyunsaturated fat diet (currently recommended by several health agencies) or (2) low polyunsaturated fat diet (representative of that consumed by a large segment of the North American population) improved antitumor immune defense and inhibited tumor growth. Rats were fed one of four semi-purified diets (20% w/w fat) for 21 days pre- and 17 days post- R3230AC mammary tumor implantation. The polyunsaturated to saturated fatty acid (P/S) ratio was either 1 (high P/S diet) or 0.35 (low P/S diet). At each P/S ratio, diets provided long-chain n-3 fatty acids at 0 or 5% w/w of total fat. Long-chain n-3 fatty acids fed in a high P/S diet did not affect tumor growth or host immune responses. In contrast, feeding long-chain n-3 fatty acids in a low P/S diet increased natural killer cell cytotoxicity, splenocyte nitric oxide and interleukin-2 production, and the proportion of activated (CD25+) CD8+ and CD28+ cells, but did not significantly inhibit tumor growth. For both P/S diets, tumor cells from rats fed long-chain n 3 fatty acids had a higher n-3 content and n-3/n-6 ratio in phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol. Furthermore, the magnitude of increase in n-3 fatty acid incorporation into tumor phospholipids was greater when fed in a low P/S diet. We demonstrated that the dietary P/S ratio significantly influences the effect of long-chain n-3 fatty acids on host immune responses and n-3 fatty acid incorporation into tumor cells. These findings warrant further consideration when designing dietary recommendations. PMID- 12088118 TI - Phase III randomized trial of droloxifene and tamoxifen as first-line endocrine treatment of ER/PgR-positive advanced breast cancer. AB - PURPOSE: This trial was designed to demonstrate equivalence between droloxifene 40 mg/d and tamoxifen 20 mg/d as first-line treatment in pre- and post-menopausal women with ER+ and/or PgR+ advanced breast cancer based on time to disease progression and tumor response. MATERIALS AND METHODS: One thousand three hundred fifty four women with measurable disease, previously untreated by hormonal or chemotherapy for advanced or recurrent breast cancer, were enrolled by 179 institutions in 35 countries. Patients were stratified at baseline for menopausal status. Patients receiving adjuvant hormonal therapy within I year were excluded. All patients gave written informed consent, were randomized to 40mg droloxifene or 20 mg tamoxifen daily as single-agent therapy and underwent tumor assessment every 3 months. A central committee reviewed digitized images for all cases of tumor progression or objective response. RESULTS: The hazard ratio (droloxifene/tamoxifen) for the primary endpoint, time to disease progression, was 1.287 favoring tamoxifen (95% C.I.: 1.114-1.487; p <.001). The objective response rate (CR+PR) was 22.4% for droloxifene and 28.6% for tamoxifen (p = .02). Tamoxifen was superior to droloxifene overall, among both pre- and postmenopausal patients and among patients < or =65 years; there was no difference among women >65 years. The hazard ratio for all-cause mortality was 0.871 (95% C.I.: 0.672-1.129; p = .29), favoring droloxifene but not statistically significant. CONCLUSIONS: Droloxifene was significantly less effective than tamoxifen overall and particularly among women under 65 years. Tamoxifen and droloxifene were both less effective in pre-menopausal women with receptor-positive disease compared to post-menopausal women. Further clinical development of droloxifene was stopped. PMID- 12088119 TI - Stereotactic large core needle biopsy for all nonpalpable breast lesions? AB - BACKGROUND: Stereotactic large-core needle biopsy (SLCNB) is a minimally invasive method for histological diagnosis of nonpalpable breast disease. We studied differences in cancer prevalence between a group of women referred through the national screening program and a non-screening group, and assessed whether the validity of SLCNB differed between these groups. METHODS: A group of non selective, consecutive patients presenting with a nonpalpable mammographic lesion, who participated in a recently conducted multicenter study regarding the accuracy of SLCNB in The Netherlands, were the basis for this study. Prevalence of carcinoma, predictive value of a benign diagnosis, sensitivity, and specificity rate of SLCNB were compared between the two groups. RESULTS: Of the 1029 lesions in 972 patients included, 858 were evaluable. In 850/858 lesions (99.1%) the reason for referral was clear. The prevalence of cancer in the screening group (n = 511 lesions) was 64.0% (95%CI 59.8-68.2), versus 49.6% in the non-screening group (n = 339) (95%CI 44.2-54.9). Respective predictive values of a benign diagnosis on SLCNB were 97.0 versus 94.8% (non-significant). The sensitivity rates of SLCNB were 98.5% (screening; 95%CI 96.5-99.5) versus 95.2% (non-screening; 95%CI 90.8-97.9). Specificity rates were 97.8 (95%CI 94.5-99.4) and 99.4% (95%CI 96.8-100), respectively. CONCLUSION: Despite a significant difference in the prevalence of carcinoma, the accuracy of SLCNB did not show a statistically significant difference between both patient groups. Therefore, SLCNB appears accurate in diagnosing nonpalpable breast lesions both in screening and non-screening patient groups. PMID- 12088120 TI - One year follow-up of women opting for presymptomatic testing for BRCA1 and BRCA2: emotional impact of the test outcome and decisions on risk management (surveillance or prophylactic surgery). AB - Genetic testing enables women at risk for hereditary breast and/or ovarian cancer to find out whether they have inherited the gene mutation (BRCA1/BRCA2), and if so, to opt for frequent surveillance and/or prophylactic surgery (bilateral mastectomy and/or oophorectomy). Here, a follow-up is described for 63 healthy women at 50% risk of being a BRCA1/BRCA2 mutation carrier who underwent genetic testing. The course of distress and problems regarding body image and sexuality up to 1 year after disclosure of the test-outcome were described separately for mutation carriers undergoing mastectomy (n = 14), for mutation carriers opting for surveillance (n = 12) and for non-mutation carriers (n = 37). Furthermore, we analyzed whether women opting for prophylactic mastectomy differed from those opting for close surveillance with respect to biographical characteristics, experiences with cancer in relatives and personality. Women opting for prophylactic mastectomy had significantly higher distress levels than mutation carriers who opted for surveillance, and the non-mutation carriers. This difference in levels of distress was highest at pre- and post-test and had almost disappeared at 1-year follow-up. Besides, mutation carriers opting for prophylactic mastectomy were more often in their thirties, more often had young children and had a longer awareness of the genetic nature of cancer in the family than those opting for regular surveillance. Adverse effects were observed in women who underwent prophylactic mastectomy (mostly in combination with immediate breast reconstruction) regarding the perception of how their breast region looked like and felt, the intimate relationship and physical wellbeing whereas women opting for prophylactic mastectomy reported more distress than the other women in the study, their distress levels had significantly decreased 6 months or longer after surgery, possibly due to the significant risk reduction of developing breast cancer. This might explain, why most women who underwent prophylactic mastectomy were satisfied with this decision, despite a perceived negative impact on body image, the intimate relationship and physical wellbeing. PMID- 12088121 TI - Repulsion or attraction? Group membership and assumed attitude similarity. AB - Three studies investigated group membership effects on similarity-attraction and dissimilarity-repulsion. Membership in an in-group versus out-group was expected to create initially different levels of assumed attitude similarity. In 3 studies, ratings made after participants learned about the target's attitudes were compared with initial attraction based only on knowing target's group membership. Group membership was based on political affiliation in Study 1 and on sexual orientation in Study 2. Study 3 crossed political affiliation with target's obnoxiousness. Attitude dissimilarity produced stronger repulsion effects for in-group than for out-group members in all studies. Attitude similarity produced greater increments in attraction for political out-group members but not for targets with a stigmatic sexual orientation or personality characteristic. PMID- 12088122 TI - The statistical analysis of data from small groups. AB - The authors elaborate the complications and the opportunities inherent in the statistical analysis of small-group data. They begin by discussing nonindependence of group members' scores and then consider standard methods for the analysis of small-group data and determine that these methods do not take into account this nonindependence. A new method is proposed that uses multilevel modeling and allows for negative nonindependence and mutual influence. Finally, the complications of interactions, different group sizes, and differential effects are considered. The authors strongly urge that the analysis model of data from small-group studies should mirror the psychological processes that generate those data. PMID- 12088123 TI - The role of public exposure in moral and nonmoral shame and guilt. AB - Although scholarly traditions assume that shame results more from the public exposure of a transgression or incompetence than guilt does, this distinction has little empirical support. Four studies, using either undergraduate participants' responses to hypothetical scenarios, their remembered experiences, or the coding of literary passages, reexamined this issue. Supporting traditional claims, public exposure of both moral (transgressions) and nonmoral (incompetence) experiences was associated more with shame than with guilt. Shame was also more strongly linked with nonmoral experiences of inferiority, suggesting 2 core features of shame: its links with public exposure and with negative self evaluation. The distinctive features of guilt included remorse, self-blame, and the private feelings associated with a troubled conscience. PMID- 12088124 TI - Place matters: consensual features and regional variation in American well-being and self. AB - Consensual and regionally distinct features of well-being and self were examined in a nationally representative survey of midlife Americans (ages 25-75). Consistent with key American ideology, Study 1 found that a majority of Americans believe they have high levels of mastery, purpose, life satisfaction, overall health, family and work obligation, and partner and family support. Study 2 found distinct regional well-being profiles (e.g., New England reflected concern with not being constrained by others; Mountain showed concern with environmental mastery; West South Central with personal growth and feeling cheerful and happy; West North Central with feeling calm, peaceful, and satisfied; and East South Central with contributing to others' well-being). Study 3 found regional self profiles consistent with the well-being profiles. PMID- 12088125 TI - Global self-esteem in relation to structural models of personality and affectivity. AB - Three studies examined global self-esteem in relation to structural models of personality and affectivity. In every study, self-esteem was strongly negatively correlated with Neuroticism/Negative Affectivity and moderately to strongly related to Extraversion/Positive Affectivity. Additional findings, however, revealed that self-esteem is better viewed at the lower order level. For instance, global self-esteem correlated -.79 with the Depression facet of the Revised NEO Personality Inventory (P. T. Costa, Jr., & R. R. McCrae, 1992) in Study 3. Moreover, confirmatory factor analyses produced very strong correlations between self-esteem and depression in both Study 2 (r = -.82) and Study 3 (r = .86). Taken together, the data suggest that global self-esteem measures define one end of a bipolar continuum, with trait indicators of depression defining the other. PMID- 12088126 TI - Episodic and semantic knowledge in emotional self-report: evidence for two judgment processes. AB - Three studies involving 3 participant samples (Ns = 39, 55, and 53) tested the hypothesis that people retrieve episodic emotion knowledge when reporting on their emotions over short (e.g., last few hours) time frames, but that they retrieve semantic emotion knowledge when reporting on their emotions over long (e.g., last few months) time frames. Support for 2 distinct judgment strategies was based on judgment latencies (Studies 1 and 2) and priming paradigms (Studies 2 and 3). The authors suggest that self-reports of emotion over short versus long time frames assess qualitatively different sources of self-knowledge. PMID- 12088127 TI - Information processing in everyday life: emotion-congruent bias in mothers' reports of parent-child interactions. AB - The authors examined the role of emotions in mothers' perceptions of the parent child relationship. Ninety-nine mothers completed a measure of trait affect and then interacted with their preschool children under low- and high-stress conditions. After each interaction session, mothers rated their mood states and positive and negative aspects of parent-child behavior. To control for differences in the quality of dyadic interaction, observers also rated behavior. Across sessions, mood was a more robust predictor of mothers' perceptions than was trait affect. When trait effects did occur, they were mediated by mood states. Results suggest that when emotionally congruent information processing occurs in real-life situations, emotions generated by the interactions play a preeminent role in the immediate interpretation of the exchange. PMID- 12088128 TI - Attaining personal goals: self-concordance plus implementation intentions equals success. AB - The present article includes separate meta-analyses showing that self-concordance and implementation intentions are significantly positively associated with goal progress. Study 1 confirmed the positive relations of both self-concordance and implementation intentions to weekend goal progress. Study 2 confirmed the positive relation of self-concordance with monthly progress on New Year's resolutions but failed to find a direct benefit for implementation intentions. Both studies, however, obtained a significant interaction effect indicating that goal self-concordance and implementation intentions combined synergistically to facilitate goal progress. The article also reports a meta-analysis and results from the 2 studies that demonstrated that goal progress was associated with improved affect over time. PMID- 12088129 TI - What is the central feature of extraversion? Social attention versus reward sensitivity. AB - R. E. Lucas, E. Diener, A. Grob, E. M. Suh, and L. Shao (2000) recently argued that the core of the personality dimension of Extraversion is not sociability but a construct called reward sensitivity. This article accepts their argument that the mere preference for social interaction is not the central element of Extraversion. However, it claims that the real core of the Extraversion factor is the tendency to behave in ways that attract social attention. Data from a sample of 200 respondents were used to test the 2 hypotheses with comparisons of measures of reward sensitivity and social attention in terms of their saturation with the common variance of Extraversion measures. The results clearly showed that social attention, not reward sensitivity, represents the central feature of Extraversion. PMID- 12088130 TI - To belong or not to belong, that is the question: terror management and identification with gender and ethnicity. AB - The terror management prediction that reminders of death motivate in-group identification assumes people view their identifications positively. However, when the in-group is framed negatively, mortality salience should lead to disidentification. Study 1 found that mortality salience increased women's perceived similarity to other women except under gender-based stereotype threat. In Study 2, mortality salience and a negative ethnic prime led Hispanic as well as Anglo participants to derogate paintings attributed to Hispanic (but not Anglo American) artists. Study 3 added a neutral prime condition and used a more direct measure of psychological distancing. Mortality salience and the negative prime led Hispanic participants to view themselves as especially different from a fellow Hispanic. Implications for understanding in-group derogation and disidentification are briefly discussed. PMID- 12088131 TI - Math = male, me = female, therefore math not = me. AB - College students, especially women, demonstrated negativity toward math and science relative to arts and language on implicit measures. Group membership (being female), group identity (self = female), and gender stereotypes (math = male) were related to attitudes and identification with mathematics. Stronger implicit math = male stereotypes corresponded with more negative implicit and explicit math attitudes for women but more positive attitudes for men. Associating the self with female and math with male made it difficult for women, even women who had selected math-intensive majors, to associate math with the self. These results point to the opportunities and constraints on personal preferences that derive from membership in social groups. PMID- 12088132 TI - The role of Afrocentric features in person perception: judging by features and categories. AB - Four studies were conducted to test the hypothesis that group-related physical features may directly activate related stereotypes, leading to more stereotypic inferences over and above those resulting from categorization. As predicted, targets with more Afrocentric features were judged as more likely to have traits stereotypic of African Americans. This effect was found with judgments of African Americans and of European Americans. Furthermore, the effect was not eliminated when a more sensitive measure of categorization processes (category accessibility) was used or when the judgement context made category distinctions salient. Of additional interest was the finding that category accessibility independently affected judgment, such that targets who could be more quickly categorized as group members were judged more stereotypically. PMID- 12088133 TI - The effects of diffuse and distinct affect. AB - In a series of suboptimal priming studies, it was shown that both affective and nonaffective reactions to a stimulus may occur without awareness. Moreover, it was demonstrated that affective information is detected earlier than nonaffective information. Therefore, early reactions to an affect-laden stimulus (e.g., a smiling man) are cognitively unappraised and thus diffuse (e.g., "positive"), whereas later affective reactions can be more specific and distinct (e.g., "a smiling man"). Through variations of prime exposure (extremely short, moderately short) the impact of early diffuse and late distinct affect on judgment was investigated. Findings show that distinctness (and prime-target similarity) is an essential determinant of whether the effect of affect is null, assimilation, or contrast. Furthermore, whether affect priming activates diffuse or distinct reactions is a matter of a fraction of seconds. PMID- 12088134 TI - The psychological bases of ideology and prejudice: testing a dual process model. AB - The issue of personality and prejudice has been largely investigated in terms of authoritarianism and social dominance orientation. However, these seem more appropriately conceptualized as ideological attitudes than as personality dimensions. The authors describe a causal model linking dual dimensions of personality, social world view, ideological attitudes, and intergroup attitudes. Structural equation modeling with data from American and White Afrikaner students supported the model, suggesting that social conformity and belief in a dangerous world influence authoritarian attitudes, whereas toughmindedness and belief in a competitive jungle world influence social dominance attitudes, and these two ideological attitude dimensions influence intergroup attitudes. The model implies that dual motivational and cognitive processes, which may be activated by different kinds of situational and intergroup dynamics, may underlie 2 distinct dimensions of prejudice. PMID- 12088135 TI - When the face reveals what words do not: facial expressions of emotion, smiling, and the willingness to disclose childhood sexual abuse. AB - For survivors of childhood sexual abuse (CSA), verbal disclosure is often complex and painful. The authors examined the voluntary disclosure-nondisclosure of CSA in relation to nonverbal expressions of emotion in the face. Consistent with hypotheses derived from recent theorizing about the moral nature of emotion, CSA survivors who did not voluntarily disclose CSA showed greater facial expressions of shame, whereas CSA survivors who voluntarily disclosed CSA expressed greater disgust. Expressions of disgust also signaled sexual abuse accompanied by violence. Consistent with recent theorizing about smiling behavior, CSA nondisclosers made more polite smiles, whereas nonabused participants expressed greater genuine positive emotion. Discussion addressed the implications of these findings for the study of disclosure of traumatic events, facial expression, and the links between morality and emotion. PMID- 12088136 TI - Effects of school-level norms on student substance use. AB - This study examines the relationship between school norms of substance use disapproval (disapproval by the student body) and students' use of cigarettes, alcohol, and marijuana. Data came from nationally representative samples of 8th (N = 16,051), 10th (N = 13,251), and 12th (N = 8,797) grade students, attending 150, 140, and 142 schools, respectively. These students participated in the Monitoring the Future Project in 1999. Measures of school norms of disapproval of substance use were obtained by aggregating students' personal disapproval of daily cigarette use, heavy drinking, and marijuana use within each school. Analysis using logistic nonlinear hierarchical models indicated that in general, school-level disapproval lowered the probability of students' use of these substances, controlling for their own disapproval and for student and school demographic characteristics. The beneficial effect of school-level disapproval of cigarette and marijuana use on 8th-grade students' probability of daily cigarette use and marijuana use was significantly higher than it was for the 12th-grade students. The effect of school-level disapproval of heavy drinking on the probability of students' drinking was not significantly different across the three grades. Further, a school environment of disapproval was also found to create a protective environment for those students in the 8th and 10th grades who were themselves not disapproving of daily cigarette use. These results argue for prevention programs that include creation of an overarching environment of disapproval of substance use in schools. PMID- 12088137 TI - A multilevel analysis of neighborhood context and youth alcohol and drug problems. AB - Various sources of data were used to examine hypothesized relations among neighborhood variables and youth alcohol and drug problems. Family members (N = 1186) were from 55 neighborhoods: 57% female, 41% African American, and 59% White. Data were clustered by neighborhood and analyzed within a multilevel design. At the neighborhood level, the study examined relations among poverty, stores selling alcohol, neighborhood social cohesion, neighborhood problems with youth alcohol and drug use, and drug and alcohol arrests. At the individual level, gender, ethnicity, adult versus child status, neighborhood social cohesion, and neighborhood problems were examined. Results indicated that more stores sold alcohol in higher poverty neighborhoods, which was associated with less social cohesion. Lower social cohesion was related to greater perceived neighborhood problems with youth alcohol and drug use, which was positively related to neighborhood youth drug and alcohol arrests. The study showed significant variation across neighborhoods and demonstrates the utility of combining different sources of neighborhood data to examine relations of interest. PMID- 12088138 TI - Alcohol, tobacco, and drug use among young adults prior to entering the military. AB - This study examined the prevalence and correlates of alcohol, tobacco, and drug use among military recruits prior to enlistment. As part of a 5-year longitudinal study, a baseline survey was conducted with 2,002 Naval recruits. Despite being overwhelmingly underage, 75% of recruits consumed alcohol in the year before enlistment, and 26% had engaged in heavy drinking. About half had used tobacco, and 31% had used drugs. Multivariate logistic regression analysis revealed that although men were not at significantly greater risk than were women for preenlistment alcohol, heavy drinking or drug use, they were at elevated risk for preenlistment tobacco use (Odds Ratio = 1.71; 95% Confidence Interval = 1.21, 2.41). Normative beliefs concerning best friend's drinking were highly predictive of each outcome. Prior use of each substance was significantly associated with prior use of other substances. Preenlistment drinking, smoking, and drug use are prevalent behaviors in this population. These data can help to heighten awareness among employers as to the prevalence of substance use in the 17-24-year-old age group, and thereby inform policy and prevention programs. PMID- 12088139 TI - Early elementary school intervention to reduce conduct problems: a randomized trial with Hispanic and non-Hispanic children. AB - Children's aggressive behavior and reading difficulties during early elementary school years are risk factors for adolescent problem behaviors such as delinquency, academic failure, and substance use. This study determined if a comprehensive intervention that was designed to address both of these risk factors could affect teacher, parent, and observer measures of internalizing and externalizing problems. European American (n = 116) and Hispanic (n = 168) children from 3 communities who were selected for aggressiveness or reading difficulties were randomly assigned to an intervention or no-intervention control condition. Intervention families received parent training, and their children received social behavior interventions and supplementary reading instruction over a 2-year period. At the end of intervention, playground observations showed that treated children displayed less negative social behavior than controls. At the end of a 1-year follow-up, treated children showed less teacher-rated internalizing and less parent-rated coercive and antisocial behavior than controls. The study's limitations and implications for prevention are discussed. PMID- 12088141 TI - Progress toward poliomyelitis eradication--Pakistan and Afghanistan, January 2000 April 2002. AB - Since 1988, when the World Health Assembly resolved to eradicate poliomyelitis worldwide, the estimated global incidence of polio has decreased 99%. Pakistan began polio eradication activities in 1994 and Afghanistan in 1997. Although polio remains endemic in the two countries, both the incidence and the geographic distribution of poliovirus have been reduced substantially. This report summarizes progress toward eradicating polio in Pakistan and Afghanistan during January 2000-April 2002. Both countries aim to stop transmission of poliovirus by the end of 2002; however, the unstable security situation in the region might threaten this success. PMID- 12088140 TI - Positive impact of competence skills and psychological wellness in protecting inner-city adolescents from alcohol use. AB - Research has shown that competence enhancement prevention programs for substance use are effective in reducing alcohol use and other problem behaviors. However, less is known about the mechanisms by which high competence helps youth avoid negative outcomes. This study explored whether greater competence is associated with increased levels of psychological wellness that in turn deters subsequent alcohol use. Specifically, 1,459 students attending 22 middle and junior high schools in New York City completed surveys that included measures of competence (decision making, self-efficacy), psychological wellness, and alcohol use. Students completed surveys at baseline, 1-year follow-up, and 2-year follow-up. Data collectors administered the questionnaire following a standardized protocol during a regular 40-min class period. On the basis of a longitudinal structural equation model, adolescents who were highly competent reported greater psychological wellness, which was then associated with less drinking. These findings highlight the potential of alcohol prevention programs designed to enhance competence and psychological wellness. PMID- 12088142 TI - Update: Rashes among schoolchildren--27 states, October 4, 2001-June 3, 2002. AB - Since October 2001, a total of 27 states has reported investigations of multiple groups of schoolchildren who have developed rashes. Rash illnesses among schoolchildren in 14 states were reported in March; since the initial report, rashes have been reported in 13 additional states (Alabama, Alaska, Illinois, Iowa, Kansas, Kentucky, Maine, Maryland, Massachusetts, Minnesota, Missouri, New Hampshire, and New Jersey). Rashes also have been reported among schoolchildren in Canada. The investigations have not identified a common source for the reported cases of rashes among U.S. schoolchildren. This report summarizes available data on these rashes and provides examples for three states. CDC is continuing to monitor reports of rashes and is providing technical assistance to state and local health departments investigating these reports. PMID- 12088143 TI - Cancer death rates--Appalachia, 1994-1998. AB - Cancer is the second leading cause of death in the United States. Although descriptive analyses of mortality data are used often to identify variations by time and person, analyses that focus on regional variations are less common. Appalachia is a U.S. region with a high prevalence of risk factors for cancer (e.g., tobacco use, physical inactivity, and inadequate access to medical care). Analyses that focus on Appalachia provide valuable information for cancer control, research, and intervention. To assess the impact of cancer in Appalachia, researchers from the University of Kentucky and Pennsylvania State University, in collaboration with CDC, analyzed mortality data from CDC's National Center for Health Statistics for 1994-1998. This report summarizes the results of that analysis, which indicate elevated cancer mortality, underscoring the need for ongoing cancer prevention and control programs as a major public health priority in this region. PMID- 12088144 TI - RNAML: a standard syntax for exchanging RNA information. AB - Analyzing a single data set using multiple RNA informatics programs often requires a file format conversion between each pair of programs, significantly hampering productivity. To facilitate the interoperation of these programs, we propose a syntax to exchange basic RNA molecular information. This RNAML syntax allows for the storage and the exchange of information about RNA sequence and secondary and tertiary structures. The syntax permits the description of higher level information about the data including, but not restricted to, base pairs, base triples, and pseudoknots. A class-oriented approach allows us to represent data common to a given set of RNA molecules, such as a sequence alignment and a consensus secondary structure. Documentation about experiments and computations, as well as references to journals and external databases, are included in the syntax. The chief challenge in creating such a syntax was to determine the appropriate scope of usage and to ensure extensibility as new needs will arise. The syntax complies with the eXtensible Markup Language (XML) recommendations, a widely accepted standard for syntax specifications. In addition to the various generic packages that exist to read and interpret XML formats, an XML processor was developed and put in the open-source MC-Core library for nucleic acid and protein structure computer manipulation. PMID- 12088145 TI - Exon-specific RNAi: a tool for dissecting the functional relevance of alternative splicing. AB - The goal of functional genomics is to determine the function of each protein encoded by an organism. Typically, this is done by inactivating individual genes and, subsequently, analyzing the phenotype of the modified organisms. In higher eukaryotes, where a tremendous amount of alternative splicing occurs, such approaches are not feasible because they have the potential to simultaneously affect multiple proteins that could have quite distinct and important functions. Thus, it is necessary to develop techniques that inactivate only a subset of proteins synthesized from genes encoding alternatively spliced mRNAs. Here we demonstrate that RNA interference (RNAi) can be used to selectively degrade specific alternatively spliced mRNA isoforms in cultured Drosophila cells. This is achieved by treating the cells with double-stranded RNA corresponding to an alternatively spliced exon. This technique may prove to be a powerful tool to assess the function of proteins synthesized from alternatively spliced mRNAs. In addition, these results have implications regarding the mechanism of RNAi in Drosophila. PMID- 12088146 TI - The MEP-1 zinc-finger protein acts with MOG DEAH box proteins to control gene expression via the fem-3 3' untranslated region in Caenorhabditis elegans. AB - Cell fates in the Caenorhabditis elegans germline are regulated, at least in part, at the posttranscriptional level. For example, the switch from spermatogenesis to oogenesis in the hermaphrodite relies on posttranscriptional repression of the fem-3 mRNA via its 3' untranslated region (UTR). Previous studies identified three DEAH box proteins, MOG-1, MOG-4, and MOG-5, that are critical for the fem-3 3' UTR control. Here we describe MEP-1, a zinc-finger protein that binds specifically to each of these three MOG proteins and that is required for repression by the fem-3 3' UTR in vivo. To investigate its in vivo function, we generated a mep-1 deletion mutant. The mep-1 null phenotype suggests a broad role for MEP-1 in C. elegans development, as it is associated with early larval arrest. In addition, mep-1 mutants can be defective in gonadogenesis and oocyte production when derived from a heterozygous mother. We suggest that MEP-1 acts together with the MOG proteins to repress fem-3 mRNA and that it also functions in other pathways to control development more broadly. PMID- 12088147 TI - Phylogenetic analysis of the structure of RNase MRP RNA in yeasts. AB - RNase MRP is a ribonucleoprotein enzyme involved in processing precursor rRNA in eukaryotes. To facilitate our structure-function analysis of RNase MRP from Saccharomyces cerevisiae, we have determined the likely secondary structure of the RNA component by a phylogenetic approach in which we sequenced all or part of the RNase MRP RNAs from 17 additional species of the Saccharomycetaceae family. The structure deduced from these sequences contains the helices previously suggested to be common to the RNA subunit of RNase MRP and the related RNA subunit of RNase P, an enzyme cleaving tRNA precursors. However, outside this common region, the structure of RNase MRP RNA determined here differs from a previously proposed universal structure for RNase MRPs. Chemical and enzymatic structure probing analyses were consistent with our revised secondary structure. Comparison of all known RNase MRP RNA sequences revealed three regions with highly conserved nucleotides. Two of these regions are part of a helix implicated in RNA catalysis in RNase P, suggesting that RNase MRP may cleave rRNA using a similar catalytic mechanism. PMID- 12088149 TI - Autoimmune epitopes in messenger RNA. AB - Patients with systemic autoimmune disorders produce autoantibodies against sequence-specific conformational RNA epitopes on U1 snRNA, 28S rRNA, and transfer RNAs. The molecular basis for immunological reactivity with these highly abundant and stable RNAs is not understood. Here, we report the existence of discrete RNA epitopes in messenger RNAs that are generally less abundant and less stable than snRNAs and tRNAs. An iterative selection and amplification procedure using pooled autoimmune patient sera identified immunoreactive mRNA species. Following deconvolution of the pools to identify the reactive sera, several mRNAs recognized by these autoantibodies were cloned and sequenced. Detailed analysis using one particular serum indicated reactivity against the messages encoding alternative splicing factor (ASF/SF2) and calmodulin. Deletion and site-directed mutagenesis determined that an epitope recognized by this serum is located in a 17-base stem-loop structure common to both messages. This serum was then used to immunoprecipitate native mRNAs encoding ASF/SF2 and calmodulin from total HeLa cell RNA. Our results demonstrate that despite its low abundance and instability, messenger RNA is capable of reacting with autoantibodies generated during an autoimmune response. These data are consistent with direct presentation as a model to explain the generation of RNA conformation-specific autoantibodies. PMID- 12088148 TI - Modification of the universally unmodified uridine-33 in a mitochondria-imported edited tRNA and the role of the anticodon arm structure on editing efficiency. AB - Editing of tRNA has a wide phylogenetic distribution among eukaryotes and in some cases serves to expand the decoding capacity of the target tRNA. We previously described C-to-U editing of the wobble position of the imported tRNA(Trp) in Leishmania mitochondria, which is essential for decoding UGA codons as tryptophan. Here we show the complete set of nucleotide modifications in the anticodon arm of the mitochondrial and cytosolic tRNA(Trp) as determined by electrospray ionization mass spectrometry. This analysis revealed extensive mitochondria-specific posttranscriptional modifications, including the first example of thiolation of U33, the "universally unmodified" uridine. In light of the known rigidity imparted on sugar conformation by thiolation, our discovery of a thiolated U33 suggests that conformational flexibility is not a universal feature of the anticodon structural signature. In addition, the in vivo analysis of tRNA(Trp) variants presented shows a single base-pair reversal in the anticodon stem of tRNA(Trp) is sufficient to abrogate editing in vivo, indicating that subtle changes in anticodon structure can have drastic effects on editing efficiency. PMID- 12088150 TI - Identification and punctate nuclear localization of a novel noncoding RNA, Ks-1, from the honeybee brain. AB - We identified a novel gene, Ks-1, which is expressed preferentially in the small type Kenyon cells of the honeybee brain. This gene is also expressed in some of the large soma neurons in the brain and in the suboesophageal ganglion. Reverse transcription-polymerase chain reaction experiments indicated that Ks-1 transcripts are enriched in the honeybee brain. cDNA cloning revealed that the consensus Ks-1 cDNA is over 17 kbp and contains no significant open reading frames. Furthermore, fluorescent in situ hybridization revealed that Ks-1 transcripts are located in the nuclei of the neural cells, accumulating in some scattered spots. These findings demonstrate that Ks-1 encodes a novel class of noncoding nuclear RNA and is possibly involved in the regulation of neural functions. PMID- 12088151 TI - Dynamic conformational model for the role of ITS2 in pre-rRNA processing in yeast. AB - Maturation of the large subunit rRNAs includes a series of cleavages that result in removal of the internal transcribed spacer (ITS2) that separates mature 5.8S and 25/28S rRNAs. Previous work demonstrated that formation of higher order secondary structure within the assembling pre-ribosomal particle is a prerequisite for accurate and efficient pre-rRNA processing. To date, it is not clear which specific sequences or secondary structures are required for processing. Two alternative secondary structure models exist for Saccharomyces cerevisiae ITS2. Chemical and enzymatic structure probing and phylogenetic comparisons resulted in one structure (Yeh & Lee, J Mol Biol, 1990, 211:699-712) referred to here as the "hairpin model." More recently, an alternate folded structure was proposed (Joseph et al., Nucleic Acids Res, 1999, 27:4533-4540), called here the "ring model." We have used a functional genetic assay to examine the potential significance of these predicted structures in processing. Our data indicate that elements of both structural models are important in efficient processing. Mutations that prevent formation of ring-specific structures completely blocked production of mature 25S rRNA, whereas those that primarily disrupt hairpin elements resulted in reduced levels of mature product. Based on these results, we propose a dynamic conformational model for the role of ITS2 in processing: Initial formation of the ring structure may be required for essential, early events in processing complex assembly and may be followed by an induced transition to the hairpin structure that facilitates subsequent processing events. In this model, yeast ITS2 elements may provide in cis certain of the functions proposed for vertebrate U8 snoRNA acting in trans. PMID- 12088152 TI - Characterization of interactions among the Cef1p-Prp19p-associated splicing complex. AB - Schizosaccharomyces pombe (Sp) Cdc5p and its Saccharomyces cerevisiae (Sc) ortholog, Cef1p, are essential components of the spliceosome. In S. cerevisiae, a subcomplex of the spliceosome that includes Cef1p can be isolated on its own; this has been termed the nineteen complex (Ntc) because it contains Prp19p. Components of the Ntc include Cef1p, Snt309p, Syf2p/Ntc31p, Ntc30p/lsy1p, Ntc20p and at least six unidentified proteins. We recently identified approximately 30 proteins that copurified with Cdc5p and Cef1p. Previously unidentified S. pombe proteins in this purification were called Cwfs for complexed with five and novel S. cerevisiae proteins were called Cwcs for complexed with Cef1p. Using these proteomics data coupled with available information regarding Ntc composition, we have investigated protein identities and interactions among Ntc components. Our data indicate that Cwc2p, Prp46p, Clf1p, and Syf1p most likely represent Ntc40p, Ntc50p, Ntc77p, and Ntc90p, respectively. We show that Sc Cwc2p interacts with Prp19p and is involved in pre-mRNA splicing. Sp cwf2+, the homolog of Sc CWC2, is allelic with the previously identified Sp prp3+. We present evidence that Sp Cwf7p, an essential protein with obvious homologs in many eukaryotes but not S. cerevisiae, is a functional counterpart of Sc Snt309p and binds Sp Cwf8p (a homolog of Sc Prp19p). Further, our data indicate that a mutation in the U-box of Prp19p disrupts these numerous protein interactions causing Cef1p degradation and Ntc instability. PMID- 12088153 TI - Probing FinO-FinP RNA interactions by site-directed protein-RNA crosslinking and gelFRET. AB - The conjugative transfer of F-plasmids is repressed by a two-component system, which consists of the antisense RNA FinP and the protein FinO. FinO binds FinP, protecting it from endonucleolytic degradation and facilitating duplex formation between FinP and its complementary RNA. Here we present the results of site specific protein-RNA cross-linking and gel-based fluorescence resonance energy transfer (gelFRET) experiments used to probe the structure of a complex of FinO bound to an RNA target consisting of a duplex with 5' and 3' single-stranded tails. The crosslinking experiments reveal that an extensive, largely positively charged surface on FinO contacts RNA. The gelFRET measurements indicate that the 5' single-stranded tail of the RNA is in closer contact with much of the protein than the distal, blunt end of the RNA duplex. These data suggest that significant conformational adjustments in the protein and/or the RNA accompany complex formation. PMID- 12088154 TI - Thermodynamic and phylogenetic prediction of RNA secondary structures in the coding region of hepatitis C virus. AB - The existence and functional importance of RNA secondary structure in the replication of positive-stranded RNA viruses is increasingly recognized. We applied several computational methods to detect RNA secondary structure in the coding region of hepatitis C virus (HCV), including thermodynamic prediction, calculation of free energy on folding, and a newly developed method to scan sequences for covariant sites and associated secondary structures using a parsimony-based algorithm. Each of the prediction methods provided evidence for complex RNA folding in the core- and NS5B-encoding regions of the genome. The positioning of covariant sites and associated predicted stem-loop structures coincided with thermodynamic predictions of RNA base pairing, and localized precisely in parts of the genome with marked suppression of variability at synonymous sites. Combined, there was evidence for a total of six evolutionarily conserved stem-loop structures in the NS5B-encoding region and two in the core gene. The virus most closely related to HCV, GB virus-B (GBV-B) also showed evidence for similar internal base pairing in its coding region, although predictions of secondary structures were limited by the absence of comparative sequence data for this virus. While the role(s) of stem-loops in the coding region of HCV and GBV-B are currently unknown, the structure predictions in this study could provide the starting point for functional investigations using recently developed self-replicating clones of HCV. PMID- 12088156 TI - Awakening to the psychopharmacology of sleep and arousal: novel neurotransmitters and wake-promoting drugs. AB - Exciting new developments in the psychopharmacology of wakefulness are clarifying the neurotransmitters, pathways, and drugs that impact this important physiologic state. Selectively inducing normal wakefulness without stimulating external vigilance may lead to therapeutic benefits not only in sleep disorders but also in cognitive disorders and conditions associated with fatigue. PMID- 12088157 TI - Risk factors for completed suicide in bipolar disorder. AB - BACKGROUND: Because Chinese bipolar patients in Taiwan, unlike Western patients, exhibit low comorbidity of substance abuse disorders, this retrospective and controlled study of completed suicide in bipolar patients explored the risk period and other risk factors for such an outcome. METHOD: All acute inpatients with bipolar I disorder (DSM-IV) were followed from date of admission after January 1, 1985, until December 31, 1996, in regard to their death. The patients were followed by record linkage to the Death Certification System in Taiwan, which was issued throughout 1996. Nineteen female and 24 male patients died as a result of suicide within this period. Forty-one of 43 of the total number of patients were matched with 1 living bipolar individual (as a control subject) for age, sex, and date of admission. Demographic data, family history, and clinical characteristics were collected from the patients' medical records and were formally confirmed at every admission. RESULTS: The lifetime prevalence of alcohol/drug use disorders was 14.6% in suicide completers. Thirty suicide completers (69.8%) revealed duration of illness of at least 7 years at the time of death. The latency period from the presumed time of onset to completing suicide averaged 12.2 years. The mean age at the first suicide attempt was 31.1 years among 43 completers and 10 living controls who had ever attempted suicide. Conditional logistic regression revealed a strong association of suicide (p < .001) with the following factors: onset with mood-congruent psychotic feature (adjusted odds ratio [OR] = 0.18, 95% confidence interval [CI] = 0.04 to 0.74), positive first-degree family history of completed suicide (adjusted OR = 15.08, 95% CI = 1.39 to 163.50), and making a suicide attempt at least once in 7 years of illness (adjusted OR = 4.96, 95% CI = 1.03 to 23.83). There appeared to be no significant difference in fasting levels of serum cholesterol or blood sugar between the suicide completers and the living controls. CONCLUSION: The first 7 to 12 years subsequent to onset of affective illness and age less than 35 years may be the high-risk periods for suicide in bipolar disorder. Those bipolar disorder patients who have a first-degree family history of suicide and who have more suicide attempts (at least once in 7 years of illness) are likely to commit suicide. Symptomatology (e.g., mood congruence of psychotic features) at the time of presumed disease onset may potentially differentiate subgroups of bipolar patients with various levels of suicide risk. PMID- 12088155 TI - Gene silencing using micro-RNA designed hairpins. AB - During RNA interference (RNAi), long dsRNA is processed to approximately 21 nt duplexes, short interfering RNAs (siRNAs), which silence genes through a mRNA degradation pathway. Small temporal RNAs (stRNAs) and micro-RNAs (miRNAs) are approximately 21 nt RNAs that are processed from endogenously encoded hairpin structured precursors, and function to silence genes via translational repression. Here we report that synthetic hairpin RNAs that mimic siRNAs and miRNA precursor molecules can target a gene for silencing, and the mechanism of silencing appears to be through mRNA degradation and not translational repression. The sequence and structural configuration of these RNAs are important, and even slight modification in structure can affect the silencing activity of the hairpins. Furthermore, these RNAs are active when expressed by DNA vectors containing polymerase III promoters, opening the possibility for new approaches in stable RNAi-based loss of function studies. PMID- 12088158 TI - Effects of antidepressant treatment on the quality of daily life: an experience sampling study. AB - BACKGROUND: Although some depression trials have included quality of life (QoL) as an outcome measure, assessments were retrospective and relatively infrequent. Because QoL varies in relation to everyday experience, intensive time-sampling approaches may be useful. METHOD: The experience sampling method (ESM) was used to assess effects of antidepressant treatment on the quality of life, as measured from moment to moment in daily life (mQoL), and related aspects of daily experience. Primary care patients with a DSM-III-R/DSM-IV diagnosis of major depressive disorder were randomly assigned to imipramine (N = 32) or placebo (N = 31) treatment for 6 weeks, with possible prolongation to 18 weeks. A healthy control group (N = 22) provided normative data. RESULTS: Treatment-related increases in frequency and severity of physical complaints, including those not reported to the general practitioner as side effects, were associated with lowered mQoL; this negative association was especially strong in treatment dropouts. Despite greater clinical improvement at week 6, imipramine patients did not report greater increases than placebo patients in mean mQoL ratings. However, imipramine treatment stabilized mQoL fluctuations and led to reductions in time spent "doing nothing." Patients' decisions to prolong treatment depended on clinical improvement, mQoL changes, and specific early side effects. At 18 weeks, remitted patients still showed deficits on ESM daily life measures relative to healthy controls, even though QoL had returned to normal on retrospective measures. CONCLUSION: ESM provides new insights in the effects of antidepressant treatment on daily life experiences and should therefore be considered as a supplement to conventional instruments in clinical trials. PMID- 12088159 TI - Predominance of anger in depressive disorders compared with anxiety disorders and somatoform disorders. AB - OBJECTIVE: The object of this study was to make a comparison regarding various dimensions of anger between depressive disorder and anxiety disorder or somatoform disorder. METHOD: The subjects included 73 patients with depressive disorders, 67 patients with anxiety disorders, 47 patients with somatoform disorders, and 215 healthy controls (diagnoses made according to DSM-IV criteria). Anger measures--the Anger Expression Scale, the hostility subscale of the Symptom Checklist-90-Revised (SCL-90-R), and the anger and aggression subscales of the Stress Response Inventory--were used to assess the anger levels. The severity of depression, anxiety, phobia, and somatization was assessed using the SCL-90-R. RESULTS: The depressive disorder group showed significantly higher levels of anger on the Stress Response Inventory than the anxiety disorder, somatoform disorder, and control groups (p < .05). The depressive disorder group scored significantly higher on the anger-out and anger-total subscales of the Anger Expression Scale than the somatoform disorder group (p < .05). On the SCL 90-R hostility subscale, the depressive disorder group also scored significantly higher than the anxiety disorder group (p < .05). Within the depressive disorder group, the severity of depression was significantly positively correlated with the anger-out score (r = 0.49, p < .001), whereas, in the somatoform and anxiety disorder groups, the severity of depression was significantly positively correlated with the anger-in score (somatoform disorder: r = 0.51, p < .001; anxiety disorder: r = 0.57, p < .001). CONCLUSION: These results suggest that depressive disorder patients are more likely to have anger than anxiety disorder or somatoform disorder patients and that depressive disorder may be more relevant to anger expression than somatoform disorder. PMID- 12088160 TI - Differential effects of nefazodone and cognitive behavioral analysis system of psychotherapy on insomnia associated with chronic forms of major depression. AB - BACKGROUND: The antidepressant nefazodone and the Cognitive Behavioral Analysis System of Psychotherapy (CBASP) were recently found to have significant, additive effects in a large multicenter study of chronic forms of major depression. As nefazodone-mediated blockade of serotonin-2 receptors may directly relieve insomnia associated with depression, we examined the more specific effects of CBASP and nefazodone, singly and in combination, on sleep disturbances. METHOD: A total of 597 chronically depressed outpatients (DSM-III-R criteria) with at least 1 insomnia symptom were randomly assigned to 12 weeks of treatment with nefazodone (mean final dose = 466 mg/day), CBASP (mean = 16.0 sessions), or the combination (mean dose = 460 mg/day plus a mean of 16.2 CBASP sessions). Continuous and categorical insomnia outcomes, derived from standard clinician- and self-rated assessments, were compared. RESULTS: Patients receiving nefazodone (either alone or in combination with CBASP) obtained significantly more rapid and greater ultimate improvement in insomnia ratings when compared with those treated with CBASP alone. This difference was maximal by the fourth week of therapy and sustained thereafter. Combined treatment did not result in markedly better insomnia scores than treatment with nefazodone alone on most measures, although patients receiving both CBASP and nefazodone were significantly more likely (p < .001) to achieve > or = 50% decrease in insomnia severity. CONCLUSION: Despite comparable antidepressant efficacy, monotherapy with nefazodone or CBASP resulted in markedly different effects on the magnitude and temporal course of insomnia symptoms associated with chronic forms of major depression. Patients receiving the combination of psychotherapy and pharmacotherapy benefited from both the larger and more rapid improvements in insomnia associated with nefazodone therapy and the later-emerging effects of CBASP on the overall depressive syndrome. PMID- 12088161 TI - A double-blind placebo-controlled study of the efficacy and safety of paroxetine in the treatment of pathological gambling. AB - BACKGROUND: This randomized, double-blind, placebo-controlled study investigated the efficacy and tolerability of paroxetine in the treatment of pathological gambling. METHOD: Patients fulfilling DSM-IV criteria for pathological gambling and scoring > or = 5 on the South Oaks Gambling Screen were enrolled if no other Axis I disorder was present. A 1-week placebo run-in phase was followed by 8 weeks' treatment with paroxetine or placebo. The initial paroxetine dose of 20 mg/day could be increased after week 2 by 10 mg/week to a maximum of 60 mg/day. Changes in clinical status were assessed using the Gambling Symptom Assessment Scale (G-SAS) and the Clinical Global Impressions scale (CGI). Treatment-emergent symptoms were assessed weekly. RESULTS: Forty-five patients were included in an intent-to-treat analysis (N = 23 paroxetine, N = 22 placebo). Statistically significantly greater reductions in the total score of the G-SAS were observed in the paroxetine group compared with the placebo group at weeks 6 through 8 (p = .003, .003, and .042, respectively). Improvement on the CGI was also significantly greater in the paroxetine than in the placebo group at the same timepoints (p = .033, .014, and .025, respectively). A significantly greater proportion of patients in the paroxetine group were responders at weeks 7 and 8 (p = .011 and .010, respectively). CONCLUSION: The results of this trial indicate that paroxetine may be effective in the treatment of pathological gambling. There were no unexpected side effects from this treatment. However, additional studies with larger patient samples and a longer treatment phase are required to establish conclusively the efficacy and safety of paroxetine for this indication. PMID- 12088162 TI - A randomized trial comparing paroxetine and venlafaxine in the treatment of bipolar depressed patients taking mood stabilizers. AB - BACKGROUND: The treatment of depressive episodes occurring in bipolar patients taking mood stabilizers is an understudied area of research with outstanding clinical consequences. This study was aimed to assess and compare the efficacy and safety of 2 different antidepressant drugs, paroxetine and venlafaxine, in this indication. METHOD: Sixty DSM-IV bipolar patients. each presenting with a major depressive episode while receiving mood stabilizers, were randomly assigned to either paroxetine (N = 30) or venlafaxine (N = 30) for 6 weeks in a single blind manner. They had to score higher than 17 on the 17-item Hamilton Rating Scale for Depression (HAM-D-17) and have their mood stabilizer blood levels within the therapeutic range. Efficacy was measured by the HAM-D. Reports of side effects were collected at each visit; switch to mania or hypomania was specifically assessed by the Young Mania Rating Scale at 5 of 7 visits. RESULTS: Significant improvements in HAM-D scores were observed in both paroxetine- and venlafaxine-treated patients (Wilcoxon p < .0001). There were no significant differences in either efficacy or safety measures between the 2 drugs. By intention-to-treat analysis, 43% (N = 13) of patients taking paroxetine and 48% (N = 14) taking venlafaxine were considered to be responders. Only 3% (N = 1) of patients switched to hypomania or mania in the paroxetine group, whereas 13% (N = 4) switched in the venlafaxine group. CONCLUSION: Paroxetine and venlafaxine are both effective and safe in the treatment of depressive breakthrough episodes in bipolar disorder. There was a suggestion of a slightly higher risk for switch to mania or hypomania with venlafaxine. PMID- 12088163 TI - Quetiapine for psychosis in Parkinson's disease versus dementia with Lewy bodies. AB - BACKGROUND: Most clinicians perceive psychosis in dementia with Lewy bodies (DLB) as more difficult to treat than Parkinson's disease, yet there are no reports comparing the antipsychotic response between the 2 disorders. METHOD: All charts of Parkinson's disease and DLB patients at our Movement Disorders Center, Memorial Hospital of Rhode Island, Pawtucket, given quetiapine for psychosis were reviewed. Demographic data, including type and severity of psychosis, before and after Unified Parkinson's Disease Rating Scale (UPDRS)-motor scores, motor worsening, and treatment response (recorded as poor/none, partial, or total), were obtained. The chi-square test was used to assess differences in efficacy and tolerability of quetiapine between Parkinson's disease and DLB patients. RESULTS: Eighty-seven Parkinson's disease and 11 DLB patients with psychosis were analyzed. No significant difference in mean age, levodopa dose, quetiapine dose, duration of quetiapine use, or baseline UPDRS-motor score was noted between Parkinson's disease and DLB patients. Eighty percent (70/87) of Parkinson's disease patients and 90% (10/11) of DLB patients had partial to complete resolution of psychosis using quetiapine (p = .40). Motor worsening was noted at one point in 32% (28/87) of Parkinson's disease and 27% (3/11) of DLB patients over the duration of quetiapine use (p = .74). CONCLUSION: Long-term quetiapine use was generally well tolerated in this geriatric Parkinson's disease and DLB population. Mild motor worsening occurred in some patients. No significant difference in long-term efficacy and motor worsening associated with quetiapine treatment was noted between the 2 disorders. PMID- 12088164 TI - A 28-week comparison of ziprasidone and haloperidol in outpatients with stable schizophrenia. AB - BACKGROUND: Ziprasidone is a novel antipsychotic with a unique pharmacologic profile. This study compared ziprasidone with the conventional antipsychotic haloperidol in outpatients with stable schizophrenia. METHOD: Three hundred one outpatients with stable chronic or subchronic schizophrenia (DSM-III-R) were randomized and participated in this double-blind, multicenter, parallel-group clinical study comparing flexible-dose oral ziprasidone, 80-160 mg/day (N = 148), with haloperidol, 5-15 mg/day (N = 153), over 28 weeks. Patients were assessed using the Positive and Negative Syndrome Scale (PANSS), the Clinical Global Impressions-Severity of Illness scale, the Montgomery-Asberg Depression Rating Scale, the Simpson-Angus Scale, the Barnes Akathisia Scale, and the Abnormal Involuntary Movement Scale. RESULTS: Modal doses at endpoint were 80 mg/day for ziprasidone and 5 mg/day for haloperidol. Improvements in all mean efficacy variables with both ziprasidone and haloperidol were observed. Significantly more patients were categorized as negative symptom responders (> or = 20% reduction in PANSS negative subscale score) in the ziprasidone group (48%) compared with the haloperidol group (33%) (p < .05). Ziprasidone had clear advantages over haloperidol in all evaluations of movement disorders. Changes in body weight were negligible with both treatments. No pattern of laboratory or cardiovascular changes was observed. CONCLUSION: Ziprasidone and haloperidol were both effective in reducing overall psychopathology; ziprasidone demonstrated effective treatment of negative symptoms and was better tolerated than haloperidol. Ziprasidone appears to offer an effective alternative to haloperidol in the long-term treatment of stable outpatients with schizophrenia. PMID- 12088165 TI - Immediate effect of intravenous diazepam in neuroleptic-induced acute akathisia: an open-label study. AB - BACKGROUND: Neuroleptic-induced akathisia can be severely distressing to some patients. and rapid treatment would be preferable. However, there have been relatively few studies conducted regarding the rapid treatment of akathisia. The effect of intravenous diazepam at the beginning of treatment for akathisia was studied in an open clinical trial. METHOD: The subjects were 18 patients with schizophrenia or bipolar I disorder (DSM-IV criteria) who developed neuroleptic induced acute akathisia during antipsychotic medication and who required immediate relief from the distress of akathisia. Diazepam was given intravenously to the patients at a rate of 5 mg per 30 seconds. RESULTS: All 18 subjects experienced immediate relief from akathisia after the injection of diazepam (mean +/- SD dose = 12.6 +/- 2.6 mg; range, 10-17 mg). They reported no serious adverse effects. CONCLUSION: The results suggest that intravenous diazepam could be used in the treatment of patients with severely distressing akathisia who require immediate relief. PMID- 12088167 TI - Fluoxetine-induced sexual dysfunction reversed by loratadine. PMID- 12088166 TI - Prevalence of obesity and weight change during treatment in patients with bipolar I disorder. AB - BACKGROUND: Obesity is a major public health concern in the United States and its prevalence is increasing. Individuals with bipolar disorder tend to be overweight, and their treatment may exacerbate obesity and increase the risk of concurrent medical disease in this population. METHOD: This retrospective report from the Pittsburgh Study of Maintenance Therapies in Bipolar Disorder examines the prevalence of overweight (body mass index [BMI] = 25.0-29.9) and obesity (BMI > or = 30.0) in 50 consecutive subjects with bipolar I disorder (DSM-IV) and evaluates weight change during acute treatment and the first year of maintenance treatment. RESULTS: At entry into the study, 34 (68%) of the patients in this sample with bipolar disorder were obese or overweight. The prevalence of obesity was high (16 [32%] of the 50 patients in our sample). The number of previous depressive episodes contributed to the likelihood of being overweight or obese at study entry. During the trial, most of the weight gain occurred during acute treatment rather than during maintenance treatment. During acute treatment, the amount of increase of BMI was positively related to the score on the Hamilton Rating Scale for Depression and negatively related to the score on Bech-Rafaelsen Mania Scale. There was a negative relationship between BMI and tendency to gain weight, during both acute and maintenance treatment. CONCLUSION: The high prevalence of obesity in subjects with bipolar disorder emphasizes the need for specific treatment strategies and programs for weight control for these individuals. The minimal weight gain during the maintenance phase, the relationship of acute depressive episodes to weight gain, and the fact that stabilization on maintenance medication may facilitate the participation in specific interventions for weight loss provide additional support for the practice of maintenance treatment. PMID- 12088168 TI - Rapid onset of antidepressant activity with venlafaxine. PMID- 12088169 TI - Surprising results in the study of paroxetine for generalized anxiety disorder. PMID- 12088170 TI - Design and interpretability of findings in a family study to investigate posttraumatic stress disorder. PMID- 12088171 TI - Clinical management of cardiovascular risks during treatment with psychotropic drugs. AB - The treatment of schizophrenia--as well as dementia, psychotic depression, and bipolar disorder--was greatly improved by the development of new atypical antipsychotic drugs in the past decade. However, cardiovascular and metabolic risks that are associated with some of these agents have become worrisome to clinicians and researchers. In this article, I will review psychotropic and other agents that have been associated with cardiovascular changes during treatment. Screening for cardiac vulnerability at baseline, as well as at regular intervals during treatment if the patient has certain risk factors, is important when treating patients with psychotropic agents. PMID- 12088172 TI - Mechanisms and risks of electrocardiographic QT interval prolongation when using antipsychotic drugs. AB - This article reviews cardiac electrophysiology, with a focus on the assessment of the electrocardiographically determined corrected QT (QTc) interval and its role as a marker for potentially life-threatening cardiac arrhythmias such as torsades de pointes. Presently, using the QTc interval as a surrogate for polymorphic ventricular tachycardia is handicapped, in part, by the limitations of currently available group-derived formulas to estimate the QTc interval. Regulatory agencies have sharpened their interest in this arena. Substantial progress almost certainly awaits the application of individual rather than group-derived formulas to estimate the QTc interval. Until this refinement arrives, clinicians are advised to exercise caution when administering antipsychotic drugs with the potential to significantly prolong the QT interval. Caution is particularly urged in patients with cardiovascular disease or risk factors for cardiovascular disease. PMID- 12088173 TI - Assessing cardiovascular risks versus clinical benefits of atypical antipsychotic drug treatment. AB - The atypical antipsychotic drugs are a major advance in the treatment of psychosis in spite of concerns about metabolic and cardiovascular side effects that affect morbidity and mortality. Concerns about weight gain, hypoglycemia, diabetes, and increases in lipids as well as sudden death due to torsades de pointes and other cardiovascular events can temper enthusiasm about the atypical antipsychotics. The challenge for the clinician is to weigh the benefits and risks for each drug for each patient and develop a treatment plan with the individual patient in mind. This article discusses both risks and benefits of antipsychotic treatment and presents a treatment algorithm to aid the clinician in choosing medications for the psychotic patient. PMID- 12088174 TI - Risk of cardiovascular disease and sudden death in schizophrenia. AB - Patients with schizophrenia are at higher risk for medical illnesses than people in the general population. Electrocardiographic and metabolic abnormalities that occur in patients with schizophrenia who are treated with antipsychotic medications have raised concerns about the safety of these medications. Concerns are amplified by the increased risk of atherosclerosis and sudden cardiac death in patients with schizophrenia regardless of the effect of antipsychotic agents. Because the modifiable risk factors for coronary atherosclerosis and sudden death are so prevalent within the schizophrenic population, it is important for clinicians treating patients with schizophrenia to know what these risks are and understand how they can contribute to increased mortality in these patients. The increased risk of atherosclerosis and sudden death in the schizophrenic population highlights a need for preventive services, which is further underscored by the numerous system- and patient-related barriers to preventive treatment. Clinicians must not only be aware of the modifiable risk factors, but they must also learn to manage the obstacles to prevention in conjunction with other health care specialists. PMID- 12088175 TI - Red blood cell count in short-term prediction of cardiovascular disease incidence in the Gubbio population study. AB - OBJECTIVE: The Gubbio Study is an Italian population study measuring risk factors and incidence for major cardiovascular diseases. This analysis investigates the association between red blood cell (RBC) count, after preliminarily taking into account haematocrit, and incidence of coronary and cardiovascular events. METHODS: A population sample of 2,469 men and women aged 35-74 years, free from major cardiovascular diseases and in whom RBC count and haematocrit were measured in 1983 along with other standard risk factors, were followed up for 6 years and incidence was estimated for both fatal and non-fatal coronary heart disease (CHD) and all cardiovascular atherosclerotic (CVD) events. Proportional hazards models were solved for the prediction of these events. RESULTS: In six years 61 CHD hard criteria, 109 CHD any criterion and 149 CVD events were recorded. Preliminarily, both haematocrit and RBC count, two highly correlated variables, were studied to predict CVD events; however, haematocrit did not contribute multivariately, in the overall population and separately in men and women. Age-adjusted rates per 1,000 of the 3 event categories were computed in sex-specific RBC count quintiles (Q) and a difference was observed between Q5 and Q1 (with 5.21 +/- 0.31 and 4.18 +/- 0.23 x 10(6) per microl, respectively) for CHD any criterion (p < 0.07) and CVD (p < 0.05). P on trends was < 0.05 for both end-points. In multivariate models, adjusted for 7 other risk factors, RBC count contributed a weak statistical significance to predict CVD incidence [relative risk (RR) for a 0.5 x 10(6) per microl difference 1.23 with 95% confidence intervals (CI) 1.00- 1.51], whereas its contribution to predict CHD any criterion (RR = 1.19 with CI 0.93- 1.51) and CHD hard criteria (RR = 1.15 with Cl 0.83-1.58) was not statistically significant. Inclusion of blood glucose and presence of diuretics (11.33% of the population) as possible confounders had no major effect although the latter were, as expected, a significant risk factor (RR = 1.90 with Cl 1.28-2.82) which further diluted the CVD predictive role of RBC count (RR = 1.22 with CI 0.99- 1.50). CONCLUSIONS: Increased RBC count is independently (yet weakly) associated with risk of CVD events in the 6-year follow-up of the Gubbio Study. Longer follow-up is needed before the contributory role of RBC count can be properly assessed to explain CHD incidence. PMID- 12088176 TI - Do differences in baseline characteristics or in management account for the poorer in-hospital and subsequent outcome after myocardial infarction in diabetics? Data from a large unselected cohort. AB - OBJECTIVE: There is an excess mortality after myocardial infarction in diabetics, but also documented significant differences in the characteristics of MI and in management between diabetics and non-diabetics. The aim of this prospective study in a large unselected patient cohort in a single French region was to determine if baseline characteristics, management, or in-hospital and one-year mortality differed in diabetic and non-diabetic patients with myocardial infarction. METHODS AND RESULTS: Data were prospectively collected in consecutive patients with myocardial infarction admitted to all hospitals in three departments in the Rhone-Alpes region between September 1, 1993 and January 31,1995. Among the 2,297 patients, 410 patients (17.8%) were diabetic. Although diabetics were older than non-diabetics (70.3 vs. 67.8 years; p < 0.0004), and less likely to receive thrombolysis (31% vs. 36%; p = 0.043), in-hospital mortality was not significantly higher (17.3% vs. 14.7%) than in non-diabetics. In multivariate analysis, diabetes was a significant predictor of one-year mortality (relative risk: 1.41; 95% CI = 1.10 - 1.79; p = 0.0063) but not of in-hospital mortality (relative risk: 1.2; 95% CI = 0.9 - 1.7; p = 0.25). Multivariate predictors of in hospital and one-year mortality in diabetics were age and Killip class at admission. CONCLUSIONS: In this large unselected French cohort, diabetes mellitus was a significant predictor of one-year but not of in-hospital mortality after myocardial infarction in a French region. This negative effect of diabetes on mortality was not related to differences in baseline characteristics, or in initial or post-discharge management between diabetics and non-diabetics. PMID- 12088177 TI - Diagnostic value of CRP and Lp(a) in coronary heart disease. AB - OBJECTIVES: Increased lipoprotein (a) [Lp(a)] concentration was reported to be an independent risk factor for coronary heart disease (CHD). Recent epidemiological studies affirmed the value of C-reactive protein (CRP) as the strongest, univariate predictor of the cardiovascular events. We decided to establish cut off levels providing maximum diagnostic efficiency for CHD. METHODS: In this study we measured CRP and Lp(a) concentrations in patients with angiographically demonstrated CHD (group A, n: 120), patients without any angiographically demonstrable lesion (group B, n: 62) and a group of healthy subjects (group C, n: 41). Data were evaluated correcting for lipid and lipoprotein concentrations, diabetes mellitus, hypertension, smoking, age, and body mass index in men and women. ROC curve based cut-off values (comparing group A versus groups B and C) and associated diagnostic performances of the assays were evaluated. RESULTS: Significant increases were noted in serum CRP concentrations in men and women, in groups A vs. B,A vs. C, B vs. C. Lp(a) concentrations were not different among groups in men but were higher in group A vs. B and C in women. Optimal cut-off levels for CRP in women and men were found as 2.1 and 3.0 mg/l with the diagnostic values of 0.792 and 0.770, respectively. For Lp(a) optimal cut-off levels were found as 22.6 and 9.8 mg/dl with the diagnostic values of 0.612 and 0.596 in women and men, respectively. CONCLUSION: The CRP level is quite efficient for separation of patients from controls. Therefore keeping in mind the lack of specificity, the CRP level may be a useful tool in the diagnosis of coronary heart disease. However, the Lp(a) level is not efficient enough to support the use of Lp(a) measurement for management of coronary heart disease. PMID- 12088178 TI - Clinical predictors of early left ventricular thrombus formation in acute myocardial infarction. AB - OBJECTIVE: Clinical epidemiological and echocardiographic risk factors relating to the development of a left ventricular thrombus were studied retrospectively in patients with acute myocardial infarction. METHODS AND RESULTS: The data on 1833 consecutive patients treated for acute myocardial infarction during a 10-year period were processed retrospectively. Transthoracic echocardiography was performed on each patient 65.0 +/- 5.5 hours after hospital admission. A left ventricular thrombus was detected in 145 patients (7.9%). The patients with acute myocardial infarction and a left ventricular thrombus had significantly lower frequencies of 1) myocardial infarction in their family history (3% versus 11%, respectively), 2) hospital admission within 24 hours from the onset of chest pain (17% versus 50%, respectively), 3) thrombolytic therapy (8% versus 23%, respectively) and 4) current smoking (24% and 35%, respectively) than those without a left ventricular thrombus. In contrast, anterior infarction (81% versus 38%, respectively), left ventricular dilatation (30% versus 19%, respectively), dyskinesis of the left ventricular wall (23% versus 10%, respectively), an aneurysm (22% versus 7%, respectively) and a reduced systolic left ventricular function (ejection fraction < 40%) (28% versus 17%, respectively) were more frequent in the presence of a left ventricular thrombus after myocardial infarction. Multivariate analysis of the results revealed that the presence of anterior myocardial infarction and an aneurysm is associated with significantly increased hazard ratios. On the other hand, early hospitalization and a positive family history of infarction significantly lowered the hazard ratio. The frequency of a left ventricular thrombus was significantly higher in spring and winter. CONCLUSIONS: The results presented in this paper confirm the significant hazard of certain parameters [location of infarction (anterior) and aneurysm] as concerns left ventricular thrombus formation among patients with acute myocardial infarction. Early hospitalization was found to lower the risk of thrombus formation. These echocardiographic and clinical parameters may be useful in the establishment of the individual risk of intracavital thrombus formation and may be of help in everyday medical practice. PMID- 12088179 TI - Systemic inflammation, Chlamydia pneumoniae DNA in circulating leukocytes and coronary atherosclerosis. AB - OBJECTIVE: Earlier studies have suggested that C. pneumoniae may be involved in the progression of atherosclerosis by contributing to the pathogenesis of inflammation in the vessel wall. The aim of the present study was to determine the prevalence of C. pneumoniae DNA in circulating white blood cells of patients with ischaemic heart disease and to correlate these findings with the extent of coronary atherosclerosis and serum markers of inflammation. METHODS AND RESULTS: In 203 consecutive patients undergoing diagnostic coronary angiography for different coronary syndromes, presence of C. pneumoniae DNA in circulating leukocytes could not be demonstrated by the polymerase chain reaction. Serum concentrations of CRP were significantly higher in patients with significant coronary artery disease compared to those with normal coronary arteries. In addition, patients with a three-vessel disease had significantly higher serum CRP compared to patients with diffuse, non-critical coronary atherosclerosis. A positive correlation was found between serum fibrinogen and serum CRP. CONCLUSION: In spite of a significant relation between serum CRP and the extent of atherosclerotic coronary artery disease, we were unable to detect C. pneumoniae DNA in circulating white blood cells. This observation suggests that there is no relation between circulating C. pneumoniae, systemic inflammation and extent of coronary atherosclerosis. PMID- 12088180 TI - Noncompaction of ventricular myocardium, complete atrioventricular block and minor congenital heart abnormalities: case report of an unusual coexistence. AB - Ventricular noncompaction is a rare cardiomyopathy characterized by numerous, prominent ventricular trabeculations and deep intertrabecular recesses and is caused by a disorder of endomyocardial morphogenesis. It can occur in patients with other congenital anomalies, such as various ventricular obstructive lesions, or in isolated form in the absence of associated anomalies. Patients with this unique disease may present with heart failure, systemic emboli, and ventricular arrhythmias. In this case report, we present an unusual coexistence of ventricular noncompaction and atrial septal aneurysm and mitral valve cleft in a young patient who presented with complete AV-block. PMID- 12088181 TI - Internal cardioversion: a worthwhile alternative after failed external cardioversion in obese patients. AB - Transthoracic external cardioversion is, if carried out properly, a highly effective cardioversion modality for most patients with atrial fibrillation. Failure to return to sinus rhythm (even for one or two beats) can be related to high transthoracic impedance. Internal cardioversion may offer a safe and worthwhile alternative for these patients, in need of sinus rhythm. This technique, mostly requiring only mild sedation, is actually performed in many electrophysiology centers. Because recurrence rates of atrial fibrillation seem to be the same for both methods, this invasive technique should be considered in patients with a rather low chance of atrial fibrillation recurrences. We present a case report of an extremely obese woman, resistant to external cardioversion. She successfully underwent internal low-energy cardioversion. PMID- 12088182 TI - Communication between left main stem trunk and proximal segment of left anterior descending coronary artery rescues left ventricular ejection fraction of a patient with three-vessel disease. AB - The well-known congenital anomalies of the left main stem refer mainly to its anomalous origin from the pulmonary artery or the right aortic sinus. We present a patient with a large communication between the left main stem trunk and the proximal segment of the left anterior descending coronary artery which preserves the left ventricular systolic performance in the setting of significant three vessel disease. PMID- 12088183 TI - The ageing male: demographics and challenges. AB - In the year 2000, there were more than 400 million people aged 65 and over in the world--projected to increase to almost 1.5 billion by the year 2050--a close to fourfold increase compared to the 50% increase for the global population as a whole. More than 25% of these 1.5 billion elderly people are projected to be "oldest-old" (aged 80 and over). Global ageing is a triumph and a challenge. As we enter the 21st century, it will put increased economic and social demands in all countries. But if more and more individuals reach older age in good health- and remain healthy for longer--the benefits will be shared by all. Therefore, the promotion of healthy ageing and the prevention of disability in all older people must assume a central role in medical care and research as well as in the formulation of national health and social policies. Effective programs promoting healthy ageing will ensure a more efficient use of health and social services and improve the quality of life in older persons by enabling them to remain independent and productive. With prolonged life expectancy, men and women can expect to live one-third of their lives with some form of hormone deficiency. Life expectancy differences between men and women exist in various regions of the world with a mean of 4.2 years, and is projected to increase to 4.8 years by the year 2050. The ageing male, in particular, has the risk of developing gender specific urological diseases, such as prostate cancer, benign prostate hyperplasia continence disorders (generally ignored by men) and erectile dysfunction. Hormonal changes in the ageing male are associated with changes in the body mass index, osteoporosis, and sleep and mood disorders. A significant relationship between body fat mass and both cardiovascular and overall mortality in men has been demonstrated. In some populations, at least, men may run a higher risk for cardiovascular complications than women. It is our sincere hope that the next few years will enrich us with facts and clarify the state of our present knowledge, permit us to recognize some of the missing links, give us the tools and methodology to design and plan ways to understand ageing in men, allow us to help to improve the quality of life, prevent the preventable, and postpone and decrease the pain and suffering of the inevitable. PMID- 12088184 TI - Diagnostic approach to the aging male. AB - Quality of life has become a major issue in health care, particularly in old age. The aging process runs its own course but certain elements potentially lend themselves to intervention. The urologist has excellent opportunities to counsel aging men on general aspects of health and to offer screening for common ailments of old age. It is relevant to distinguish between the process of aging itself and diseases occurring in old age. The latter should be diagnosed and adequately treated. Risk factors for these age-related diseases should be identified and addressed. Several questionnaires have been developed that are helpful. Apart from urological disease, cardiovascular disease, diabetes mellitus and osteoporosis are common conditions. There are now tests available that allow screening. Of great importance is the need to raise awareness in aging men of the possibilities modern medicine offers to improve quality of life, and to encourage these men to utilize them for their benefit. PMID- 12088185 TI - Ageing, hormones, body composition, metabolic effects. AB - The age-associated, progressive changes in body composition are paralleled by a progressive decrease in hormone levels, among which there are testosterone (T) and growth hormone (GH). Moreover, the age-associated changes are rather similar to the changes in body composition seen in hypogonadism or hyposomatotropism, respectively. These age-associated changes in body composition are rather important: a decline in muscle mass by 20%-40% between the ages of 25 and 75, a doubling of fat mass and a decrease in bone mineral density by 0.3% per year after age 35. They have important functional and metabolic consequences. Most studies show a significant positive correlation between muscle mass and bone mineral density, respectively, and plasma T or GH levels, whereas abdominal fat mass is generally negatively correlated with T and GH levels. Whether, however, the hormone levels play a causal role or are the consequence of the changes in body composition is still an unsettled problem. The beneficial effects of hormone substitution on body composition suggest, nevertheless, that T and GH are at least co-determinants of the observed age-associated changes. PMID- 12088186 TI - The ageing male and erectile dysfunction. AB - Erectile dysfunction (ED) is common in the ageing man and reliable therapies are needed. The pathophysiology of erectile dysfunction in this patient group mainly includes chronic ischaemia, which triggers the deterioration of cavernous smooth muscle and the development of corporeal fibrosis. The assessment of the ageing man with erectile dysfunction who seeks medical treatment should include a thorough medical and sexual history, a systemic and focused physical examination and selected blood tests. An extensive discussion of the various therapeutic options should follow in order to allow the patient to identify the most suitable alternative. Oral drug therapy represents a safe and efficacious option for most elderly patients. PMID- 12088187 TI - The ageing penis. AB - Epidemiological reports have demonstrated the determinant role that age plays in the pathophysiological mechanism of an erection. The atherosclerosis of the penis and its ischaemia that occurs with ageing impacts directly on the intracavernous structures and the erectile physiologic function. The effects of testosterone on libido and sexual behaviour are well established but its function in the erectile mechanism remains unclear. In the animals, testosterone deficiency seems to provoke a decrease in smooth muscle content and nitric oxide (NO) production. In human, testosterone seems to affect the central system rather than the peripheral mechanism of an erection. The role of ischaemia and the decrease in testosterone in the mechanism of erectile dysfunction are reviewed. PMID- 12088188 TI - The ageing male. AB - With prolonged life expectancy, men and women can expect to live one-third of their lives with some form of hormone deficiency. The ageing male, in particular, has the added problem of developing urological diseases, such as benign prostatic hyperplasia (BPH), prostate cancer, continence disorders and erectile dysfunction. When discussing age-related problems, it is often difficult to separate and to distinguish between the natural ageing process, ageing amplifiers and an acute or chronic illness, or inter-current diseases. Partial endocrine deficiencies of ageing are associated with a decrease in the peripheral levels of testosterone, dehydroepiandrosterone (DHEA), DHEA sulphate (DHEA-S), growth hormone, insulin-like growth factor and melatonin. There is also a concomitant increase in luteinising hormone and follicle stimulating hormone. The concentration of free biologically active testosterone is lowered further by an increase in sex hormone binding globulin (SHBG). Hormonal changes in the ageing male are associated with changes in the body mass index, osteoporosis, sleep and mood disorders. A number of testosterone replacement therapies are available. These therapies should maintain physiological levels not only of serum testosterone, but also of its metabolites, including dihydrotestosterone (DHT) and estradiol. Men on testosterone therapy should be monitored at 3-month intervals during the first year of use and, thereafter, at 1-year intervals if they are stable. The association of testosterone replacement with development of prostate cancer has not been determined. PMID- 12088189 TI - Some historical reflections on the ageing male. AB - Throughout the early history of medicine, discourses on the "ageing male" can be found, although most authors addressed the aspect of old age rather than the early onset and beginnings of this period of life. With reference to the typical climacteric changes in women, the term "climacteric disease" in males was coined by H. Halford in 1813 and finally reintroduced into modern medical terminology in the 1930s by A. A. Werner. In between, several authors have described this clinical entity, often called "climacterium virile", by considering hormonal and neurological changes to be the underlying causes. All these early writings should be of certain interest to those who are dealing with modern aspects of the ageing male and are reflected in this article. PMID- 12088190 TI - Local antiseptic therapy for non-gonococcal urethritis: phase II study with Instillagel. AB - Non-gonococcal urethritis (NGU) is conventionally treated with oral antibiotics. With this Phase II study, we investigated the action of a locally disinfecting substance, Instillagel, in symptomatic NGU. Instillation treatment was performed twice daily to 32 male patients with symptomatic NGU. To evaluate the therapeutic outcome, a smear was taken from the urethra and an urine examination was performed at baseline as well as at 5 and 8 days after the start of the treatment. Pain and micturition symptoms were determined by a questionnaire with analog scales taken before and after therapy. A pre/post comparison of the urethral smears of the patients with symptomatic NGU showed a significant difference (P < 0.0001). The microbial count in the urine did not show a significant difference. The symptoms micturition (P<0.0001) and pain in the urethra (P<0.0001) were significantly improved. This Phase II study confirmed that local antiseptic treatment of NGU can offer an alternative to systemic antibiotic treatment. PMID- 12088191 TI - The endoscopic extraperitoneal radical prostatectomy (EERPE): technique and initial experience. AB - Using the experiences of the extraperitoneal (endoscopic pelvic lymphadenectomy and inguinal hernia repair) and the transperitoneal approach (laparoscopic radical prostatectomy), we developed a totally extraperitoneal approach to endoscopic radical prostatectomy. In view of the favourable short-term outcome, we describe the technique of totally extraperitoneal endoscopic radical prostatectomy (EERPE) as a now standardised procedure. After creating the preperitoneal space by balloon dissection, five trocars were placed in the hypogastrium, allowing immediate access to the space of Retzius. The surgical technique of EERPE replicates the steps of the classical retropubic descending radical prostatectomy with slight modifications. The procedure starts with exposing the Retzius space and pelvic lymph node dissection. After that, the endopelvic fascia and the puboprostatic ligaments are incised, followed by ligating the Santorini plexus. The actual prostate dissection is similar to the open descending approach: bladder neck dissection, freeing of the seminal vesicles, transsectioning of the prostatic vesicles (with or without preserving the neurovascular bundles) and, finally, apical dissection. A water-tight urethrovesical anastomosis is performed with interrupted sutures. There were 20 patients who underwent EERPE. Mean operating time was 170 min with no conversion. No major complications occurred. Only one patient required a blood transfusion. The catheter could be removed on postoperative day 6 (n = 17) or on postoperative day 12 (n = 3). Final pathologic evaluations were 4 stage pT2a, 10 stage pT2b, 5 stage pT3a, and 1 pT3b. Surgical margins were negative in 17 patients. By avoiding entry into the peritoneal cavity, therefore, obviating intra-abdominal complications, such as bowel injury, ileus, or intestinal adhesions, the extraperitoneal endoscopic access provides a safe and minimally invasive approach to the prostate, combining the advantages of minimally invasive laparoscopy and retropubic open prostatectomy. PMID- 12088192 TI - Modified pump-driven venous bypass in surgery for renal cell carcinoma (RCC) involving the inferior vena cava (IVC). AB - Surgical management of renal cell carcinoma (RCC) with a tumour thrombus that infiltrates the caval wall or extends above the hepatic veins can be problematic. Total control of the suprahepatic inferior vena cava (IVC) is mandatory in order to prevent thrombus mobilization and minimize blood loss. Pump-driven veno-venous bypass (VVB), modified by adding portal decompression, is a safe and useful procedure and avoids the important risks connected with deep hypothermic circulatory arrest while allowing the normal perfusion of vital organs. PMID- 12088193 TI - Systemic and cavernous plasma levels of vasoactive intestinal polypeptide during sexual arousal in healthy males. AB - Results from basic research scrutiny indicate a role for non-adrenergic, non cholinergic transmitters, among which there are various peptides, in the physiology of normal male sexual function. Nevertheless, it is not yet known which particular peptides are essentially involved in maintaining sexual arousal and regulating penile tumescence and rigidity in adult males. Vasoactive intestinal polypeptide (VIP), a peptide with smooth muscle relaxing properties, is considered to be one of the factors that contributes to such control. The present study was performed to evaluate the significance of VIP in normal male sexual function. We determined the plasma levels of VIP in the systemic and cavernous blood of 54 healthy adult male volunteers, who were exposed to visual and tactile erotic stimuli in order to elicit penile tumescence and erection. Whole blood was aspirated from the corpus cavernosum and the cubital vein during penile flaccidity, tumescence, rigidity and detumescence, and VIP was quantified in plasma aliquots by means of a radioimmunoassay. Of the 54 volunteers, 16 were permitted to masturbate and ejaculate, and blood was then again withdrawn from the cavernous meshwork and the cubital vein in order to measure VIP. All VIP levels were registered within the normal physiological range from 3.0-30 pmol/l. No increase in median VIP plasma levels was observed in the systemic and cavernous blood when the flaccid penis became rigid. During penile detumescence, mean cavernous VIP level increased to 11.9+/-7.8 pmol/l (baseline: 8.6+/-3.0 pmol/l), whereas VIP remained unaltered in the systemic circulation. Following ejaculation, mean VIP level in the cavernous blood was elevated to 25.3+/-10.9 pmol/l, whereas, in the systemic blood, no significant changes were registered. Our results support the hypothesis that VIP plays a functional role in the mechanism of male sexual arousal. Nevertheless, our data indicate that the peptide is not the main non-adrenergic, non-cholinergic mediator of penile tumescence and rigidity in human males. PMID- 12088194 TI - The effect of N-acetyl-L-cysteine on the viscosity of ileal neobladder mucus. AB - N-acetyl-L-cysteine (NAC) proved to be an effective mucolytic in pulmonary secretions. Our goal was to investigate the in vitro effect of NAC on viscosity of ileal neobladder mucus. The urine of a patient with an ileal neobladder was collected during the first 7 days postoperatively and stored in a refrigerator. After precipitation, the urine was decanted. The residue was stirred to a homogeneous suspension. To samples of 4.5 ml mucus, 0.5 ml NAC 10% was added. To the control sample, 0.5 ml water was added. The samples were incubated in a water bath at 37 degrees C for 5, 30 and 60 min. Viscosity was measured in the Bohlin VOR Rheometer. The viscosity of the ileal neobladder mucus decreased quickly after incubating with NAC 10%. Viscosity increased slightly after I h of incubation. The viscosity in the control sample was higher than in the other incubated samples. NAC was found to decrease the viscosity of ileal neobladder mucus, supporting the in vivo experience that NAC can be useful in patients with an ileal neobladder to facilitate the evacuation of mucus by decreasing viscosity. PMID- 12088195 TI - Neurapraxic complications in operations performed in the lithotomy position. AB - In this study, we assessed the incidence and risk factors associated with lower extremity neurapraxia in operations performed in the lithotomy position. A total of 1170 patients, who were operated on in the lithotomy position, were evaluated retrospectively. During the post-operative neurological evaluation, those patients who developed neuropraxic complications were reviewed in detail to identify those with lower extremity sensory and motor deficiencies. The electromyographic (EMG) results of the patients with neurological deficiencies were evaluated. The patients were studied with respect to age, length of operation, type of operation and mode of anaesthesia. Fischer's Exact Probability Test was used for the statistical analyses. Post-operative neurapraxia complications developed in 12 of the patients (1.02%). Of these cases, two had irreversible neurological deficiencies. All other patients recovered without any treatment during the first post-operative month. No correlation was discovered between the type of anaesthesia used and neurapraxia (P> 0.05). However, it was found that age, type of operation and operation time contributed to neurapraxia developing (P < 0.05). Old age (older than 70), prolonged operation time (more than 180 min) and an inappropriate lithotomy position were seen as the main risk factors in developing post-operative neurapraxia. Mistakes in the positioning of the operating staff might be an additional contributing factor. PMID- 12088196 TI - Carcinogenicity and mechanism of action of fumonisin B1: a mycotoxin produced by Fusarium moniliforme (= F. verticillioides). AB - Fumonisins are fungal metabolites and suspected human carcinogens. They inhibit ceramide synthase in vitro, enhance tumor necrosis factor alpha (TNFalpha) production, and cause apoptosis. Fumonisin B1 (FB1) was fed to rats and mice for 2 years or, in separate studies, given to rats or mice for up to 4 weeks. Kidney tubule adenomas and carcinomas were found in male rats fed > or = 50 ppm, whereas liver adenomas and carcinomas were found in female mice fed > or = 50 ppm for 2 years. In the short-term studies, increases in tissue concentration of the ceramide synthase substrate sphinganine (Sa) and the Sa to sphingosine (So) ratio were correlated with apoptosis. Further, hepatotoxicity was ameliorated in mice lacking either the TNFR1 or the TNFR2 TNFalpha receptors. Thus, FB1 was carcinogenic to rodents and thefindings support the hypothesis that disrupted sphingolipid metabolism and TNFalpha play important roles in its mode of action. PMID- 12088197 TI - Effect of nucleotide substitutions in N-acetyltransferase-1 on N-acetylation (deactivation) and O-acetylation (activation) of arylamine carcinogens: implications for cancer predisposition. AB - Genetic polymorphism in N-acetyltransferase-1 (NAT1) is associated with increased risk of various cancers, but epidemiological investigations are compromised by poor understanding of the relationship between NAT1 genotype and phenotype. Human reference NAT1*4 and 12 known human NAT1 allelic variants possessing nucleotide polymorphisms in the NAT1 coding region were cloned and expressed in yeast (Schizosaccharomyces pombe). Large reductions in the N-acetylation of 4 aminobiphenyl and the O-acetylation of N-hydroxy-2-aminofluorene were observed for recombinant NAT1 allozymes encoded by NAT1*14B, NAT1*15, NAT1*17, NAT1*19, and NAT1*22. Each of these alleles exhibited substantially lower expression of NAT1 protein than the reference NAT1*4 and the other NAT1 alleles. These results show an important effect of the NAT1 genetic polymorphism on the N- and O acetylation of arylamine carcinogens, suggesting modification of cancer susceptibility following exposures to arylamine carcinogens. PMID- 12088198 TI - Differences in N-acetylation genotypes between Caucasians and Black South Africans: implications for cancer prevention. AB - Polymorphic N-acetyltransferase genes (NAT1 and NAT2) determine rapid or slow acetylation phenotypes, which are believed to affect cancer risk related to environmental exposure. Black South Africans have a unique incidence pattern of environment-related cancers, but genetic characteristics of this population are mostly unknown. In this study, we compared NAT1 and NAT2 allele distributions in 101 Black South Africans and 112 UK Caucasians. Frequencies of the rapid alleles were significantly higher in Black South Africans for both NAT1 and NA72. Putative rapid NAT1 genotypes due to the presence of either NAT1*10 or NAT1*11 were found in 74.3% of Black South Africans (only NAT1*10) and 42.0% of UK Caucasians (P < 0.0001). Similarly, NAT2 analysis showed that the presence of NA12*4, NAT2*12A, NAT2*12B, NA72*12C, and NAT2*13 alleles provided significantly higher (P = 0.0001) frequency of rapid acetylation genotypes among Black South Africans (60.4%) than in the Caucasian group (33.9%). The rapid acetylation genotype in Caucasians usually depended on the NAT2*4 allele presence. The significant differences in N-acetylation genotypes can be among the factors determining a distinctive cancer morbidity and mortality pattern observed in Black South Africans. Both further genetic characterization of different populations and development of preventive strategies adopted for ethnicities with different genetic backgrounds are needed to deal adequately with the emerging health care problems in developing multiethnic societies. PMID- 12088199 TI - Oral contraceptives and risk of familial breast cancer. AB - The risk of breast cancer of oral contraceptive (OC) use in 1423 women from families with hereditary/familial breast cancer recruited through a cancer family clinic was analyzed in a matched case-control study. Ninety-eight women tested positive for a BRCA1 mutation. Hazard ratio for ever use of OCs adjusted for other risk factors was 0.90 (95% confidence interval (CI) 0.68-1.18) in the total data set and 2.00 (0.36-10.9) in BRCA1 mutation carriers. We did not find evidence for interaction between BRCA1 mutation status and OC use on breast cancer risk. Recent users had a statistically significant increase in risk with hazard ratios of 1.99, 2.05, and 1.69 for up to 5, 10, and 15 years since last OC use, while users with more than 15 years since last use had a reduction of risk to 0.69 compared to never users. We conclude that the effects of OC use on breast cancer risk in familial breast cancer may be similar to the effects in the general population. For BRCA1 mutation carriers, the point estimate is a doubling of risk, but CI is wide and no conclusion may be drawn from this study alone. PMID- 12088200 TI - Inner-city African American women who failed to receive cancer screening following a culturally-appropriate intervention: the role of health insurance. AB - Culturally-appropriate health promotion programs are thought to be more effective among minority groups than those designed for the population at large. We investigated factors associated with failure to obtain cervical and breast cancer screening among inner-city African American women who received a culturally appropriate educational intervention. Women who completed the intervention, but did not obtain a Pap smear, a clinical breast examination, and/or a mammogram at follow-up were compared with those who did obtain these tests. Women with private health insurance were more likely to be screened following the intervention than those covered by Medicaid or Medicare or those who were not insured (P < 0.001). Post-intervention screening was not associated with age, education, income, employment, or marital status. The effectiveness of a culturally-appropriate intervention is likely to be reduced if women's ability to respond is limited by inadequate insurance coverage. PMID- 12088201 TI - Breast cancer screening: stages of adoption among Cambodian American women. AB - BACKGROUND: Little information is available on the breast cancer screening behavior of Cambodian American women. METHODS: We identified households from multiple sources using Cambodian surnames and conducted a cross-sectional survey, administered by bilingual and bicultural interviewers. Breast cancer screening stages of adoption were examined based on concepts from the transtheoretical model of behavioral change. RESULTS: Our response rate was 73% (398 women in clinical breast exam (CBE) analysis, and 248 in mammography analysis) with approximately 25% each in the maintenance stage. We found significant associations between screening stage with physician characteristics. Asian American female physician increased the likelihood of being in the maintenance stage (CBE, OR = 10.1, 95% CI 2.8-37.1; mammogram, OR = 74.7,95% CI 8.3-674.6), compared to Asian American male physician with precontemplation/contemplation stage as our referent outcome. CONCLUSION: Results from this study support the need to promote regular breast cancer screening among Cambodian American women. PMID- 12088203 TI - The active fraction of plasmatic plasminogen activator inhibitor type 1 as a possible indicator of increased risk for metastatic melanoma. AB - Plasminogen activator inhibitor type 1 (PAI1) is considered to be the main regulator of fibrinolytic activity in blood and has been identified as a key enzyme in the metastasis and vascularization of solid tumors. The aim of this study was to determine whether high or low plasma levels and/or activity of PAI1 correlate with the presence of metastatic disease for patients with melanoma. We hypothesized that the presence of metastases could result in a disturbance of the fibrinolytic balance of the blood. To test our hypothesis, we have developed a unique enzyme-linked immunosorbent assay (ELISA) that can measure both the total amount as well as the active fraction of PAI1 in the plasma. We then used this novel assay to analyze the plasmatic PAI1 levels and activity of patients with advanced melanoma (AM, n = 18) and primary melanoma (PM, n = 21) and compare it to a control population (n = 38). We found no statistically significant difference in the total plasmatic PAI1 levels between the controls and patients with PM or AM (P = 0.6199). In contrast, there was a significant difference in the active fraction of PAI1 between the controls and patients with PM or AM (P = 0.0076). The difference between the control and AM groups was highly significant (P = 0.0042). A value of less than 44% active PAI1 was shown to be clinically meaningful by linear discriminant analysis. Surprisingly, the difference between the control and PM groups was also significant--although borderline (P = 0.0488). Of the patients with PM, 19% had PAI1 activity values less than 44%, which strongly supports further investigations to determine whether plasmatic PAI1 activity might be a biological marker of increased metastatic risk. PMID- 12088202 TI - Comparison of potential biological markers cathepsin B, cathepsin L, stefin A and stefin B with urokinase and plasminogen activator inhibitor-1 and clinicopathological data of breast carcinoma patients. AB - Cysteine, serine and metalloproteinases and their respective inhibitors are involved in tumor cell invasion and may have prognostic value for the outcome of malignant disease. The aim of the study was to compare the expression of new potential biological tumor markers, the lysosomal cysteine proteinases and their endogenous inhibitors, with that of the serine proteinases and their inhibitors in breast cancinoma and to relate their levels to the clinicopathological factors of the disease. Enzyme-linked immunosorbent assays (ELISAs) were used to measure cysteine cathepsin B (CatB) and cathepsin L (CatL) and their inhibitors, stefin A (StA) and stefin B (StB), together with urokinase (u-PA) and plasminogen activator inhibitor-1 (PAI-1), in 150 cytosols of primary invasive breast carcinoma. A good correlation was found between the levels of the two cysteine proteinases but only a moderate one between those of the cysteine and serine proteinases. u-PA and PAI-1 levels correlated positively with histological grade and negatively with estrogen receptor (ER) status. PAI-1 correlated with most clinicopathological factors that indicate the progression of the disease, while cathepsins and stefins were independent of these factors. In the total group of patients, high u-PA and PAI-1 and low StB levels correlated significantly with shorter disease-free survival (DFS), while CatB, CatL and StA did not. In lymph node negative patients, high CatB and CatL were also associated with shorter DFS, while u-PA remained the most significant of all these biological markers. In conclusion, this retrospective study showed u-PA to be of better prognostic relevance than the cysteine proteinases, though CatB and CatL were relevant for prognosis in lymph node negative breast cancer patients. PMID- 12088204 TI - Detection of micrometastatic cells in breast cancer by RT-pCR for the mammaglobin gene. AB - The detection of circulating cancer cells in the bone marrow (BM) and peripheral blood (PB) of patients with solid tumors may be useful for disease staging. To this aim, we evaluated the expression of the mammaglobin gene by reverse transcriptase polymerase chain reaction (RT-PCR) in 60 patients with breast cancer. Moreover, several controls were examined to test the specificity of this marker. The positive cases included 23.6% of the patients with and 9% of those without metastasis. Only 4/60 negative controls analyzed were positive by PCR. Our results show high specificity and a good correlation with disease status. PMID- 12088206 TI - KAI1/CD82 protein expression in primary prostate cancer and in BPH associated with cancer. AB - Current prognostic methods in primary prostate cancer cannot accurately identify patients with clinically significant disease at highest risk of developing metastases. This study examined KAI1/CD82 metastasis suppressor expression by quantitative immunohistochemical analysis of benign prostatic hyperplasia (BPH) and prostate cancer specimens. Altogether, prostate cancers exhibited significant KAI1 overexpression compared to BPH not associated with cancer (P = 0.022). Increased KAI1 expression in well and moderately differentiated cancers, above levels seen in BPH, with decreased expression in poorly differentiated cancers was observed. Interestingly, KAI1 expression in BPH associated with cancers was significantly higher than in BPH not associated with cancer (P = 0.009). Thus, KAI1 overexpression may restrain onset and early stage prostate cancer development, whilst its loss may predispose the patient to more aggressive cancer behaviour. Altered KAI1 expression in prostate cancers and BPH associated with cancer may have important diagnostic roles. PMID- 12088205 TI - c-erbB-2 protein in serum of primary lung cancer patients. AB - We determined c-erbB-2 protein level in serum of 86 primary lung cancer patients (78 non-small cell lung carcinomas (NSCLC), 3 small cell carcinomas, 5 not histologically defined) and in 61 controls. Aim of this study was to evaluate the clinical usefulness of c-erbB-2 as marker for lung cancer diagnosis. The protein was measured with a commercially available sandwich enzyme immunoassay. Mean levels of c-erbB-2 were 72.8 +/- 122.3 fmol/ml in lung cancers and 64.6 +/- 17.5 fmol/ml in controls (P = 0.2). No association was found between c-erbB-2 levels and histotype, tumor stage, sex and smoking habits. Among NSCLC, only four patients showed a c-erbB-2 concentration higher than the selected cut-off value of 99.6 fmol/ml. Subjects with levels higher than the 75th percentile in tumors (73 fmol/ml) had a shorter median survival than those with lower levels (6.3 months versus 10.0 months, P = 0.003). Our results indicated that serum c-erbB-2 protein is not a reliable diagnostic marker. There is, however, a suggestion of a possible clinical usefulness in terms of survival prediction. PMID- 12088207 TI - The influence of dietary iron and tocopherols on oxidative stress and ras-p21 levels in the colon. AB - The purpose of this investigation was to determine how dietary levels of alpha tocopherol, gamma-tocopherol and iron influence oxidative stress and ras-p21 levels in the colon. Rats were fed diets deficient in tocopherols (-E) or supplemented with either 0.156 mmol of alpha-tocopherol (AE)/kg diet or 0.156 mmol of gamma-tocopherol (GE)/kg of diet. Half the rats in each of these three groups received dietary iron at a level of 35 mg/kg diet and the other half at eight times this level (280 mg/kg diet). Rats fed the AE diets had higher levels of Vitamin E in feces, colonocytes, plasma and liver than did rats fed the GE diets. Dietary iron levels did not influence tocopherol levels in plasma, liver or feces. For colonocytes, high dietary iron decreased tocopherol levels. The ratio of gamma-tocopherol (in the GE groups) to alpha-tocopherol (in the AE groups) was 0.13 for plasma, 0.11 for liver, 0.28 for colonocytes and 0.51 for feces. The plasma ratio is not, therefore, predictive of the ratio in colonocytes and feces. High levels of dietary iron increased levels of fecal lipid hydroperoxides. Moreover, rats fed the GE diets had lower levels of fecal lipid hydroperoxides than rats fed the AE diets. The levels of ras-p21 were significantly lower in rats fed the GE diets compared with rats fed the AE diets. The gamma-tocopherol may, therefore, play a significant role in preventing colon cancer. High levels of dietary iron were found to promote oxidative stress in feces and colonocytes. PMID- 12088208 TI - Effects of bile acids on base hydroxylation in a model of human colonic mucosal DNA. AB - BACKGROUND AND AIMS: Increased intestinal bile acids as a possible consequence of a high fat/meat, low fiber diet are believed to play an important role in the formation of colon cancer. Interactions of bile salts particularly secondary bile acids with different cell components including DNA may contribute to carcinogenesis. To further investigate DNA damage by bile salts, we assessed the effects of a bile salt mixture containing deoxycholate and chenodeoxycholate on base hydroxylation in Chelex-treated DNA from calf thymus as a model of human colonic mucosal DNA in the presence and absence of reactive oxygen metabolites (ROM). METHODS: Chelex-treated DNA from calf thymus (to remove residual iron impurities) was incubated with different bile salt concentrations (4 microM, 4.0 mM) (20.0% deoxycholate, 21.0% chenodeoxycholate) in the presence and absence of an OH generating system (25 microM FeCl3, 50 microM H2O2, 100 microM nitrilotriacetic acid) for 18 h (37 degrees C). After hydrolyzation, lyophilization and derivatization hydroxylated DNA bases were characterised and quantitated with gas chromatography-mass spectrometry (GS-MS) and SIM analysis. Two concentration ranges of bile salts were used, micromolar concentrations being present in plasma, millimolar in the gut lumen. RESULTS: In the absence of ROM Chelex-treated DNA preparations contain only small amounts of hydroxylated base products. Bile salts at 4.0 mM significantly increased the amounts of 5-OH uracil and cis-thymine glycol. In the presence of ROM bile salts at 4.0 microM increased the production of 8-OH adenine and 8-OH guanine whereas bile salts at 4.0 mM inhibited ROM-induced base hydroxylation. DISCUSSION: In the absence of ROM millimolar concentrations of a bile salt mixture with deoxycholate and chenodeoxycholate increase basal (spontaneous) DNA hydroxylation, whereas, they are without effects at micromolar concentrations. In the presence of ROM micromolar concentrations enhance oxidative DNA damage and millimolar concentrations were inhibitory. These results support the view that bile acids may cause oxidative DNA damage depending on their concentrations and the surrounding conditions both directly (enhancement of basal hydroxylation) and indirectly (enhancement of ROM-induced hydroxylation). PMID- 12088209 TI - Caloric restriction mimetics: metabolic interventions. AB - Caloric restriction (CR) retards diseases and aging in laboratory rodents and is now being tested in nonhuman primates. One way to apply these findings to human health is to identify and test agents that may mimic critical actions of CR. Panel 2 focused on two outcomes of CR, reduction of oxidative stress and improved glucoregulation, for which candidate metabolic mimics exist. It was recommended that studies on oxidative stress should emphasize mitochondrial function and to test the efficacy of nitrone and other antioxidants in mimicking CR's effects. Studies should also focus on the long-term effects of compounds known to lower circulating glucose and insulin concentrations or to increase insulin sensitivity. Also, four other developing areas were identified: intermediary metabolism, response to infection, stress responses, and source of dietary fat. These areas are important because either they hold promise for the discovery of new mimetics or they need to be explored prior to initiation of CR trials in humans. Other recommendations were that transgenic approaches and adult-onset CR should be emphasized in future studies. PMID- 12088211 TI - Caloric restriction mimetics: physical activity and body composition changes. AB - As the only paradigm that has consistently increased life span and inhibited the onset and/or progression of disease, dietary restriction has multiple effects on a variety of organ systems. In this brief review, the goal of the panel was to attempt to understand the role of changes in physical activity and body composition as possible modulators of the life span in experimental animals and humans. We focus on whether changes in exercise behavior and body composition produce similar changes as those found in dietary restriction and whether these changes can be used to either replace or enhance the beneficial effects of dietary restriction. The complexity of the two stimuli is emphasized in our report, with suggestions offered on how to better interpret existing research. Our panel briefly examines evidence in experimental animals and humans about the specific contributions of each of these factors to altering life span and age related pathologies. We also discuss additional animal studies and/or human intervention studies that could be performed to clarify these issues. Finally, we provide suggested avenues for future research in this area of changes in physical activity and body composition as dietary restriction mimetics. PMID- 12088210 TI - Neuroendocrine and pharmacological manipulations to assess how caloric restriction increases life span. AB - As part of an effort to review current understanding of the mechanisms by which caloric restriction (CR) extends maximum life span, the authors of the present review were requested to develop a list of key issues concerning the potential role of neuroendocrine systems in mediating these effects. It has long been hypothesized that failure of specific neuroendocrine functions during aging leads to key age-related systemic and physiological failures, and more recently it has been postulated that physiological neuroendocrine responses to CR may increase life span. However, although the acute neuroendocrine responses to fasting have been well studied, it is not clear that these responses are necessarily identical to those observed in response to the chronic moderate (30% to 50% reduction) CR that increases maximum life span. Therefore the recommendations of this panel fall into two categories. First, further characterization of neuroendocrine responses to CR over the entire life span is needed. Second, rigorous interventional studies are needed to test the extent to which neuroendocrine responses to CR mediate the effects of CR on life span, or alternatively if CR protects the function of essential neuroendocrine cells whose impairment reduces life span. Complementary studies using rodent models, nonhuman primates, and humans will be essential to assess the generality of elucidated mechanisms, and to determine if such mechanisms might apply to humans. PMID- 12088212 TI - Human implications of caloric restriction's effects on aging in laboratory animals: an overview of opportunities for research. PMID- 12088213 TI - Genetic variability in responses to caloric restriction in animals and in regulation of metabolism and obesity in humans. AB - Panel 5 focused on genetic factors that might mediate or moderate the effects of caloric restriction (CR) on longevity. Panel members stated that currently there is limited information directly addressing these issues. Therefore, they focused attention on what studies could be done. In addition, the panel believed that certain conceptual issues merited clarification and focused attention on this issue. Human studies and studies of nonhuman model organisms were discussed. The panel found at least three reasons why it would be valuable to find genes that influence the (putative) longevity-promoting effect of CR in humans. Such knowledge would offer: (a) the ability to predict individual responses to CR; (b) increased understanding of physiological mechanisms; and (c) the potential to develop mechanism-based interventions to promote longevity or healthy aging. In addition, the panel emphasized several macro-level recommendations regarding research strategies to avoid, research strategies to emphasize, and resources needing development. PMID- 12088214 TI - Physiologic effects of lowering caloric intake in nonhuman primates and nonobese humans. AB - Caloric restriction (CR) reduces the rate of aging and increases life span in all small animal species studied to date, but the effects of CR in humans remain uncertain. This review summarizes current knowledge of the effects of CR in nonhuman primates and humans. The results suggest that CR has a range of beneficial effects in nonhuman primates studied under laboratory conditions, and short-term markers of CR seen in animal models appear to occur in humans subject to CR also. However, the overall benefit of CR in human populations remains to be established, and studies in human populations are needed. PMID- 12088215 TI - Epidemiologic data on the relationships of caloric intake, energy balance, and weight gain over the life span with longevity and morbidity. AB - Animal experiments have shown that calorically restricted (CR) animals weigh less and live longer than their ad libitum-fed peers. Are these observations applicable to human beings? This is an important question because the prevalence of obesity in America has increased markedly over recent years. We examine whether there are physiologic effects that occur with CR in humans that could plausibly explain the observed longevity of laboratory animals associated with CR. We also review epidemiologic data from observational and interventional studies on the relationships of caloric intake, energy balance, and weight gain with age-related diseases and longevity. Additionally, data on whether long-term, sustained maintenance of weight loss is feasible, as well as the degree of CR achieved in clinical trials, are summarized. Finally, we provide recommendations regarding further epidemiologic research that will help clarify unanswered questions in these areas. PMID- 12088216 TI - Assessing the risks of terrorism: a special collection of perspectives articles by former presidents of the Society for Risk Analysis. PMID- 12088217 TI - View point: risk analysis and the terrorism problem in two parts. PMID- 12088218 TI - A perspective: risk analysis as a tool for reducing the risks of terrorism. AB - The destruction by terrorists of the twin towers of the World Trade Center and major damage wrought to the Pentagon on September 11, 2001, followed closely by the bioterrorist anthrax attacks via the mails raised the question of whether risk analysis might have a place in defending the United States against terrorist attacks. After first reviewing the multifaceted nature of terrorism and the reasons it is likely to become endemic in world society in the long term, just as other areas of crime are endemic, this article surveys several fields of risk analysis, finding possible short- and long-term uses of risk analysis. The areas chiefly considered are: risk communication and chemical, biological, and technological risk analysis. Broad policy and other uses are also considered. The author finds that risk analysis has already played some role, perhaps informally, but he sees the possibility for a much larger, formal one, a role that is centrally important for the present and future of the United States and the world. PMID- 12088219 TI - A mental model proposed to address sustainability and terrorism issues. AB - I have assembled traditional ways to think about human needs and power along with empirical data to support a mental model of human values. The hierarchy of needs from the world of psychology and the hierarchy of power from the world of diplomacy provide a structure for the model. The empirical data collected from several nations over the last three decades support the structure. Furthermore, an examination of specific trends in this data for specific values indicates that it is not impossible to achieve a sustainable world driven by sustainable values. A world that will be defined by its successful movement toward the "triple bottom line," a term articulated by John Elkington, is a world in which economic prosperity, environmental protection, and social equity are aligned. To say that the model allows one to address terrorism is based on the assumption that the lack of social equity or the perception of that lack determines the likelihood of terrorism. PMID- 12088220 TI - Perspectives on the use of risk assessment to address terrorism. PMID- 12088221 TI - Terrorism as hazard: a new species of trouble. PMID- 12088222 TI - The role of insurance in managing extreme events: implications for terrorism coverage. PMID- 12088223 TI - The role of risk analysis in the protection of critical infrastructures against terrorism. PMID- 12088224 TI - Fusion of intelligence information: a Bayesian approach. AB - The attack that occurred on September 11, 2001 was, in the end, the result of a failure to detect and prevent the terrorist operations that hit the United States. The U.S. government thus faces at this time the daunting tasks of first, drastically increasing its ability to obtain and interpret different types of signals of impending terrorist attacks with sufficient lead time and accuracy, and second, improving its ability to react effectively. One of the main challenges is the fusion of information, from different sources (U.S. or foreign), and of different types (electronic signals, human intelligence. etc.). Fusion thus involves two very distinct and separate issues: communications, i.e., ensuring that the different U.S. and foreign intelligence agencies communicate all relevant and accurate information in a timely fashion and, perhaps more difficult, merging the content of signals, some "sharp" and some "fuzzy," some dependent and some independent into useful information. The focus of this article is on the latter issue, and on the use of the results. In this article, I present a classic probabilistic Bayesian model sometimes used in engineering risk analysis, which can be helpful in the fusion of information because it allows computation of the posterior probability of an event given its prior probability (before the signal is observed) and the quality of the signal characterized by the probabilities of false positive and false negative. Experience suggests that the nature of these errors has been sometimes misunderstood; therefore, I discuss the validity of several possible definitions. PMID- 12088225 TI - Conditional dose-response relationships for microorganisms: development and application. PMID- 12088226 TI - Benchmark dose calculations for methylmercury-associated delays on evoked potential latencies in two cohorts of children. AB - Delays in evoked potential latencies were observed at increased exposures to methylmercury from seafood in two cohorts of children. Because this outcome parameter appeared to be virtually independent of confounders, including cultural differences, a joint analysis of benchmark doses was carried out. Comparable cohort members included 382 Faroese and 113 Madeiran children without middle ear infection or neurological disease at age seven years. Maternal hair-mercury concentrations at parturition in the Faroese cohort ranged from 0.6 to 39.1 microg/g (geometric average, 4.49 microg/g). In Madeira, mothers who had not changed their diet since pregnancy had current hair-mercury concentrations ranging from 1.1 to 54.4 microg/g (geometric average 10.14 microg/g). The mercury associated delay in peak III latencies at two frequencies (20 and 40 Hz) showed similar regression equations in the two groups of children, and benchmark dose calculations were therefore carried out for the two groups separately and jointly. For a doubling of a 5% prevalence of abnormal results of the peak III latencies at 40 Hz in a linear dose-response model, the benchmark dose for the maternal hair-mercury concentration was 8.79 microg/g for the Faroese children; 8.04 microg/g for the Madeiran children; and 9.46 microg/g for both groups. Results were similar for the 20 Hz condition. Benchmark dose results were substantially lower using a logarithmic or square root curve function, although the difference in fit between the curves was far from statistically significant. The benchmark results using evoked potential latencies are in close agreement with results based on neuropsychological test performance. PMID- 12088228 TI - Perspectives on incorporating human neurobehavioral end points in risk assessments. AB - Three topics are addressed: (1) measurement issues (e.g., the reliability and validity of neurobehavioral test scores), (2) general principles of assessment, including test selection, and (3) interpretation of scores. Psychological tests generally perform as well as medical tests in terms of reliability and validity. Test manuals, assessment textbooks, and psychologists are useful resources to the risk assessor. The variety of different tests employed in neurobehavioral studies complicates interstudy comparisons. In addition, tests that ostensibly assess the same general domain of function might assess somewhat different abilities within that domain. Although a uniform battery for use in all studies seems desirable, the battery appropriate for a specific study depends on study goals, knowledge about the mechanism(s) of neurotoxicity, nature of the study population, and pattern of exposure. Exposure-related neurobehavioral deficits are generally indicators of "altered function" rather than of "clinical disease." Limiting concern to end points corresponding to clinical disease might not be appropriate. Many neurobehavioral diagnoses are phenomenological and a neurotoxicant might cause a unique pattern of deficits for which no label has been created. The concern that a small shift in the central tendency of a distribution of test scores has no significance for the individual should be reexamined in light of the prevention paradox, formulated on the basis of epidemiologic studies of chronic disease. Poor performance on a neurobehavioral test does not necessarily map clearly onto underlying behavioral or neural substrate. The absence of such linkages, given current knowledge about brain-behavior relationships, should not reduce confidence in neurobehavioral end points. Use of neurobehavioral test scores involves considerations that differ little from those that the risk assessor routinely addresses in using end points commonly used in research on other topics in environmental epidemiology. PMID- 12088227 TI - Cryptosporidium dose-response studies: variation between hosts. AB - The issue of variation is highly important in dose-response analysis: variation among genetically related pathogens infecting the same host, but also variation among hosts, in susceptibility to infection by the same pathogen. This latter issue is addressed here for the protozoan parasite Cryptosporidium parvum, the causative agent for many outbreaks of water-borne gastrointestinal illness. In human feeding studies, infectivity has been shown to be low in subjects with high preexisting anti-Cryptosporidium IgG-levels. Here we adapt the hit theory model of microbial infection to incorporate covariables, characterizing the immune status of the susceptible host. The probability of any single oocyst in the inoculum to cause infection appears to depend on preexisting IgG-levels. This does not necessarily imply direct protection by the humoral immune system; high IgG-levels may reflect a recent episode of infection/illness, and be an epi phenomenon associated with other protective responses. The IgG-dependence of the dose-response relation can be easily applied in quantitative risk analysis. The distribution of anti-Cryptosporidium IgG levels in the general population is accessible by analyzing serum banks, which are maintained in many Western countries. Using such an approach provides first insights into the variation of susceptibility to infection in the general population. PMID- 12088229 TI - Importance of uncertainty and variability to predicted risks from trophic transfer of PCBs in dredged sediments. AB - Biomagnification of organochlorine and other persistent organic contaminants by higher trophic level organisms represents one of the most significant sources of uncertainty and variability in evaluating potential risks associated with disposal of dredged materials. While it is important to distinguish between population variability (e.g., true population heterogeneity in fish weight, and lipid content) and uncertainty (e.g., measurement error), they can be operationally difficult to define separately in probabilistic estimates of human health and ecological risk. We propose a disaggregation of uncertain and variable parameters based on: (1) availability of supporting data; (2) the specific management and regulatory context (in this case, of the U.S. Army Corps of Engineers/U.S. Environmental Protection Agency tiered approach to dredged material management); and (3) professional judgment and experience in conducting probabilistic risk assessments. We describe and quantitatively evaluate several sources of uncertainty and variability in estimating risk to human health from trophic transfer of polychlorinated biphenyls (PCBs) using a case study of sediments obtained from the New York-New Jersey Harbor and being evaluated for disposal at an open water off-shore disposal site within the northeast region. The estimates of PCB concentrations in fish and dietary doses of PCBs to humans ingesting fish are expressed as distributions of values, of which the arithmetic mean or mode represents a particular fractile. The distribution of risk values is obtained using a food chain biomagnification model developed by Gobas by specifying distributions for input parameters disaggregated to represent either uncertainty or variability. Only those sources of uncertainty that could be quantified were included in the analysis. Results for several different two dimensional Latin Hypercube analyses are provided to evaluate the influence of the uncertain versus variable disaggregation of model parameters. The analysis suggests that variability in human exposure parameters is greater than the uncertainty bounds on any particular fractile, given the described assumptions. PMID- 12088231 TI - The limits of catastrophe aversion. AB - We discuss the management of catastrophe-risks from a theoretical point of view. The concept of a catastrophe is informally and formally defined, and a number of desiderata for catastrophe-averse decision rules are introduced. However, the proposed desiderata turn out to be mutually inconsistent. As a consequence of this result, it is argued that the "rigid" form of catastrophe aversion articulated by, for example, the maximin rule, the maximum probable loss rule, (some versions of) the precautionary principle, and the rule proposed in Ekenberg et al. (1997, 2000) should be given up. An alternative form of "non-rigid" catastrophe aversion is considered. PMID- 12088230 TI - The Spokane Tribe's multipathway subsistence exposure scenario and screening level RME. AB - Exposure scenarios are a critical part of risk assessment; however, representative scenarios are not generally available for tribal communities where a traditional subsistence lifestyle and diet are relevant and actively encouraged. This article presents portions of a multipathway exposure scenario developed by AESE, Inc. in conjunction with the Spokane Tribal Cultural Resources Program. The scenario serves as the basis for a screening-level reasonable maximum exposure (RME) developed for the Midnite Uranium Mine Superfund site. The process used in developing this scenario balances the need to characterize exposures without revealing proprietary information. The scenario and resulting RME reflect the subsistence use of original and existing natural resources by a hypothetical but representative family living on the reservation at or near the mine site. The representative family lives in a house in a sparsely populated conifer forest, tends a home garden, partakes in a high rate of subsistence activities (hunting, gathering, fishing), uses a sweat lodge daily, has a regular schedule of other cultural activities, and has members employed in outdoor monitoring of natural and cultural resources. The scenario includes two largely subsistence diets based on fish or game, both of which include native plants and home-grown produce. Data gaps and sources of uncertainty are identified. Additional information that risk assessors and agencies need to understand before doing any kind of risk assessment or public health assessment in tribal situations is presented. PMID- 12088232 TI - Introduction to special section on sensitivity analysis and summary of NCSU/USDA workshop on sensitivity analysis. AB - This guest editorial is a summary of the NCSU/USDA Workshop on Sensitivity Analysis held June 11-12, 2001 at North Carolina State University and sponsored by the U.S. Department of Agriculture's Office of Risk Assessment and Cost Benefit Analysis. The objective of the workshop was to learn across disciplines in identifying, evaluating, and recommending sensitivity analysis methods and practices for application to food-safety process risk models. The workshop included presentations regarding the Hazard Assessment and Critical Control Points (HACCP) framework used in food-safety risk assessment, a survey of sensitivity analysis methods, invited white papers on sensitivity analysis, and invited case studies regarding risk assessment of microbial pathogens in food. Based on the sharing of interdisciplinary information represented by the presentations, the workshop participants, divided into breakout sessions, responded to three trigger questions: What are the key criteria for sensitivity analysis methods applied to food-safety risk assessment? What sensitivity analysis methods are most promising for application to food safety and risk assessment? and What are the key needs for implementation and demonstration of such methods? The workshop produced agreement regarding key criteria for sensitivity analysis methods and the need to use two or more methods to try to obtain robust insights. Recommendations were made regarding a guideline document to assist practitioners in selecting, applying, interpreting, and reporting the results of sensitivity analysis. PMID- 12088233 TI - Hazard analysis and critical control point (HACCP) history and conceptual overview. AB - The concept of Hazard Analysis and Critical Control Point (HACCP) is a system that enables the production of safe meat and poultry products through the thorough analysis of production processes, identification of all hazards that are likely to occur in the production establishment, the identification of critical points in the process at which these hazards may be introduced into product and therefore should be controlled, the establishment of critical limits for control at those points, the verification of these prescribed steps, and the methods by which the processing establishment and the regulatory authority can monitor how well process control through the HACCP plan is working. The history of the development of HACCP is reviewed, and examples of practical applications of HACCP are described. PMID- 12088235 TI - Sensitivity analysis for importance assessment. AB - We review briefly some examples that would support an extended role for quantitative sensitivity analysis in the context of model-based analysis (Section 1). We then review what features a quantitative sensitivity analysis needs to have to play such a role (Section 2). The methods that meet these requirements are described in Section 3; an example is provided in Section 4. Some pointers to further research are set out in Section 5. PMID- 12088234 TI - Identification and review of sensitivity analysis methods. AB - Identification and qualitative comparison of sensitivity analysis methods that have been used across various disciplines, and that merit consideration for application to food-safety risk assessment models, are presented in this article. Sensitivity analysis can help in identifying critical control points, prioritizing additional data collection or research, and verifying and validating a model. Ten sensitivity analysis methods, including four mathematical methods, five statistical methods, and one graphical method, are identified. The selected methods are compared on the basis of their applicability to different types of models, computational issues such as initial data requirement and complexity of their application, representation of the sensitivity, and the specific uses of these methods. Applications of these methods are illustrated with examples from various fields. No one method is clearly best for food-safety risk models. In general, use of two or more methods, preferably with dissimilar theoretical foundations, may be needed to increase confidence in the ranking of key inputs. PMID- 12088236 TI - Illustration of sampling-based methods for uncertainty and sensitivity analysis. AB - A sequence of linear, monotonic, and nonmonotonic test problems is used to illustrate sampling-based uncertainty and sensitivity analysis procedures. Uncertainty results obtained with replicated random and Latin hypercube samples are compared, with the Latin hypercube samples tending to produce more stable results than the random samples. Sensitivity results obtained with the following procedures and/or measures are illustrated and compared: correlation coefficients (CCs), rank correlation coefficients (RCCs), common means (CMNs), common locations (CLs), common medians (CMDs), statistical independence (SI), standardized regression coefficients (SRCs), partial correlation coefficients (PCCs), standardized rank regression coefficients (SRRCs), partial rank correlation coefficients (PRCCs), stepwise regression analysis with raw and rank transformed data, and examination of scatter plots. The effectiveness of a given procedure and/or measure depends on the characteristics of the individual test problems, with (1) linear measures (i.e., CCs, PCCs, SRCs) performing well on the linear test problems, (2) measures based on rank transforms (i.e., RCCs, PRCCs, SRRCs) performing well on the monotonic test problems, and (3) measures predicated on searches for nonrandom patterns (i.e., CMNs, CLs, CMDs, SI) performing well on the nonmonotonic test problems. PMID- 12088237 TI - Use of sensitivity analysis to assess reliability of metabolic and physiological models. AB - Because ethical considerations often preclude directly determining the human health effects of treatments or interventions by experimentation, such effects are estimated by extrapolating reactions predicted from animal experiments. Under such conditions, it must be demonstrated that the reliability of the extrapolated predictions is not excessively affected by inherent data limitations and other components of model specification. This is especially true of high-level models composed of ad hoc algebraic equations whose parameters do not correspond to specific physical properties or processes. Models based on independent experimental data restricting the numerical space of parameters that do represent actual physical properties can be represented at a more detailed level. Sensitivities of the computed trajectories to parameter variations permit more detailed attribution of uncertainties in the predictions to these low-level properties. S-systems, in which parameters are estimated empirically, and physiological models, whose parameters can be estimated accurately from independent data, are used to illustrate the applicability of trajectory sensitivity analysis to lower-level models. PMID- 12088238 TI - Risk and uncertainty analysis in government safety decisions. AB - Probabilistic risk analysis (PRA) can be an effective tool to assess risks and uncertainties and to set priorities among safety policy options. Based on systems analysis and Bayesian probability, PRA has been applied to a wide range of cases, three of which are briefly presented here: the maintenance of the tiles of the space shuttle, the management of patient risk in anesthesia, and the choice of seismic provisions of building codes for the San Francisco Bay Area. In the quantification of a risk, a number of problems arise in the public sector where multiple stakeholders are involved. In this article, I describe different approaches to the treatments of uncertainties in risk analysis, their implications for risk ranking, and the role of risk analysis results in the context of a safety decision process. I also discuss the implications of adopting conservative hypotheses before proceeding to what is, in essence, a conditional uncertainty analysis, and I explore some implications of different levels of "conservatism" for the ranking of risk mitigation measures. PMID- 12088239 TI - Variability and uncertainty meet risk management and risk communication. AB - In the past decade, the use of probabilistic risk analysis techniques to quantitatively address variability and uncertainty in risks increased in popularity as recommended by the 1994 National Research Council that wrote Science and Judgment in Risk Assessment. Under the 1996 Food Quality Protection Act, for example, the U.S. EPA supported the development of tools that produce distributions of risk demonstrating the variability and/or uncertainty in the results. This paradigm shift away from the use of point estimates creates new challenges for risk managers, who now struggle with decisions about how to use distributions in decision making. The challenges for risk communication, however, have only been minimally explored. This presentation uses the case studies of variability in the risks of dying on the ground from a crashing airplane and from the deployment of motor vehicle airbags to demonstrate how better characterization of variability and uncertainty in the risk assessment lead to better risk communication. Analogies to food safety and environmental risks are also discussed. This presentation demonstrates that probabilistic risk assessment has an impact on both risk management and risk communication, and highlights remaining research issues associated with using improved sensitivity and uncertainty analyses in risk assessment. PMID- 12088240 TI - Social functionalist frameworks for judgment and choice: intuitive politicians, theologians, and prosecutors. AB - Research on judgment and choice has been dominated by functionalist assumptions that depict people as either intuitive scientists animated by epistemic goals or intuitive economists animated by utilitarian ones. This article identifies 3 alternative social functionalist starting points for inquiry: people as pragmatic politicians trying to cope with accountability demands from key constituencies in their lives, principled theologians trying to protect sacred values from secular encroachments, and prudent prosecutors trying to enforce social norms. Each functionalist framework stimulates middle-range theories that specify (a) cognitive-affective-behavioral strategies of coping with adaptive challenges and (b) the implications of these coping strategies for identifying empirical and normative boundary conditions on judgmental tendencies classified as errors or biases within the dominant research programs. PMID- 12088241 TI - Toward a method of selecting among computational models of cognition. AB - The question of how one should decide among competing explanations of data is at the heart of the scientific enterprise. Computational models of cognition are increasingly being advanced as explanations of behavior. The success of this line of inquiry depends on the development of robust methods to guide the evaluation and selection of these models. This article introduces a method of selecting among mathematical models of cognition known as minimum description length, which provides an intuitive and theoretically well-grounded understanding of why one model should be chosen. A central but elusive concept in model selection, complexity, can also be derived with the method. The adequacy of the method is demonstrated in 3 areas of cognitive modeling: psychophysics, information integration, and categorization. PMID- 12088242 TI - Toward a perceptual theory of transparency. AB - Theories of perceptual transparency have typically been developed within the context of a physical model that generates the percept of transparency (F. Metelli's episcotister model, 1974b). Here 2 fundamental questions are investigated: (a) When does the visual system initiate the percept of one surface seen through another? (b) How does it assign surface properties to a transparent layer? Results reveal systematic deviations from the predictions of Metelli's model, both for initiating image decomposition into multiple surfaces and for assigning surface attributes. Specifically, results demonstrate that the visual system uses Michelson contrast as a critical image variable to initiate percepts of transparency and to assign transmittance to transparent surfaces. Findings are discussed in relation to previous theories of transparency, lightness, brightness, and contrast-contrast. PMID- 12088243 TI - A psychophysical theory of intensity proportions, joint presentations, and matches. AB - Empirically testable assumptions relate 3 psychophysical primitives: presentations of pairs of physical intensities (e.g., pure tones of the same frequency and phase to the 2 ears or 2 successive tones to both ears); a respondent's ordering of such signal pairs by perceived intensity (e.g., loudness); and judgments about 2 pairs of stimuli being related as some proportion (numerical factor, as in magnitude production). Explicit behavioral assumptions lead to 2 families of psychophysical functions, one corresponding to unbiased joint presentations and the other to biased ones. Under an invariance assumption, the psychophysical functions in the unbiased case are approximate power functions, and those in the biased case are exact power functions. A number of testable predictions are made. The mathematics involved draws from publications in utility theory and mathematics but with a reinterpretation of the primitives. PMID- 12088244 TI - Acquisition and extinction in autoshaping. AB - C. R. Gallistel and J. Gibbon (2000) presented quantitative data on the speed with which animals acquire behavioral responses during autoshaping, together with a statistical model of learning intended to account for them. Although this model captures the form of the dependencies among critical variables, its detailed predictions are substantially at variance with the data. In the present article, further key data on the speed of acquisition are used to motivate an alternative model of learning, in which animals can be interpreted as paying different amounts of attention to stimuli according to estimates of their differential reliabilities as predictors. PMID- 12088245 TI - Dynamic field theory of movement preparation. AB - A theoretical framework for understanding movement preparation is proposed. Movement parameters are represented by activation fields, distributions of activation defined over metric spaces. The fields evolve under the influence of various sources of localized input, representing information about upcoming movements. Localized patterns of activation self-stabilize through cooperative and competitive interactions within the fields. The task environment is represented by a 2nd class of fields, which preshape the movement parameter representation. The model accounts for a sizable body of empirical findings on movement initiation (continuous and graded nature of movement preparation, dependence on the metrics of the task, stimulus uncertainty effect, stimulus response compatibility effects, Simon effect, precuing paradigm, and others) and suggests new ways of exploring the structure of motor representations. PMID- 12088246 TI - Role congruity theory of prejudice toward female leaders. AB - A role congruity theory of prejudice toward female leaders proposes that perceived incongruity between the female gender role and leadership roles leads to 2 forms of prejudice: (a) perceiving women less favorably than men as potential occupants of leadership roles and (b) evaluating behavior that fulfills the prescriptions of a leader role less favorably when it is enacted by a woman. One consequence is that attitudes are less positive toward female than male leaders and potential leaders. Other consequences are that it is more difficult for women to become leaders and to achieve success in leadership roles. Evidence from varied research paradigms substantiates that these consequences occur, especially in situations that heighten perceptions of incongruity between the female gender role and leadership roles. PMID- 12088247 TI - Theory development should begin (but not end) with good empirical fits: a comment on Roberts and Pashler (2000). AB - S. Roberts and H. Pashler (2000) argued against using goodness of fit as evidence to support theories. The authors agree with their suggestions for how to go beyond good fits but disagree with their starting point. In this comment, the authors argue that good fits are part and parcel of theory development, that they are part and parcel of the processes suggested by S. Roberts and H. Pashler, and that they must be the starting point (though far from the ending point) in theoretical development. The authors discuss historical examples of scientific theory development, recent examples of psychological theory development, and development of a particular theory (social contagion theory; J. L. Rodgers & D. C. Rowe, 1993) that S. Roberts and H. Pashler criticized. PMID- 12088248 TI - "Misura" project: a retrospective survey on the use of 5fluorouracil in the treatment of colorectal cancer in 24 Italian clinical centers. AB - The "Misura" project is a retrospective survey, with the aim to evaluate how 5FU is used in the treatment of colorectal cancer in clinical practice in Italian oncology departments. Twenty-four centers participated. Patients seen in the second half of 1998 with colorectal cancer and treated with 5FU were analyzed. Observed patients were 664, 45.9% of patients presented metastatic disease. Biochemical modulation with folinic acid and bolus 5FU was the most used schedule (59%). The De Gramont (LV 5FU2) regimen, alone or with other cytotoxic drugs, was the second most chosen schedule (14%). The most frequent side effect observed was gastrointestinal toxicity. No hematological toxicity was demonstrated in 68.8% of patients. Cutaneous toxicity occurred in 21.1% of patients. 5FU is widely used independently by the stage of disease. In palliative treatment a variety of schedules were administered by the Italian centers, lacking a standard therapy. There are very few surveys investigating oncology clinical practice. A larger survey on this issue is auspicable. PMID- 12088249 TI - Quality of life in breast and colon cancer long-term survivors: an assessment with the EORTC QLQ-C30 and SF-36 questionnaires. AB - AIMS AND BACKGROUND: Although the long-term survival of cancer patients has risen during the last decades, little is known about their quality of life. Assessment of the long-term effect of disease and treatments on subjective qualitative aspects of life that are related to health may be important for identifying specific long-term sequelae of cancer and treatments. In order to better understand the outcomes on a long-term survivor population, a sample of breast and colon cancer patients was evaluated using state-of-the-art health-related quality of life (HRQoL) approaches and instruments PATIENTS AND METHODS: EORTC OLQ-C30 and SF-36 HS questionnaires were mailed to a sample of subjects with a diagnosis of breast or colon cancer recruited in two randomized clinical trials with a median follow-up of 65 and 100 months, respectively. Data obtained with the SF-36 HS questionnaire were compared with data from a representative sample of the Italian population, whereas the EORTC QLQ-C30 data were evaluated across relevant subgroups. RESULTS: Out of 2208 subjects, 1772 were randomized to receive the different types of questionnaires and 921 answered (52%). According to the findings, if no serious comorbidity or relapse appeared, long-term cancer survivors reported having a substantially satisfying HRQoL, not much different from (age and sex) comparable controls. The most intriguing findings were those obtained from breast cancer patients. They reported a somewhat higher level than comparable Italian women of limitations in activities, when describing their role, as due to physical problems. In addition, breast cancer patients also reported lower scores (worse health perception) in two EORTC scales describing activities of daily living than colon cancer patients. When data were plotted according to the type of medical therapy, hormone therapy versus chemotherapy, a higher HRQoL was reported by the former than the latter group. CONCLUSIONS: Our study supports the hypotheses that the impact of cancer on quality of life over a period of time is not necessarily devastating in survivors. As regards the impact of different medical strategies, although our data are consistent with other research supporting the hypotheses that hormone therapy does not cause a decrease in long-term quality of life, differences we observed between hormone and chemotherapy might be caused by the action of confounds we were not able to control for or by the distortion introduced by the design of the present study. PMID- 12088250 TI - Age-related differences in patients admitted to a palliative home care service. AB - AIMS: The aim of the study was to investigate possible differences in access to the service, symptomatology and therapy in relation to age among terminal cancer patients admitted to a home care program. We examined prospectively all 116 terminal cancer patients enrolled in a home care program in 1998, comparing those up to 70 years of age (48 patients) with those above 70 (68 patients). We also compared the age-related characteristics of this population with those of all 348 patients enrolled in the program in 1989-1991. There were no significant differences between the two age groups of the 1998 population in terms of symptoms, tumor site or medication, although NSAID use tended to be greater in older patients, and opioid and anti-emetic use greater in younger patients. Patients up to 70 years of age had significantly shorter survival from admission to home care than those over 70, and a greater proportion had metastases. There were no such significant age-related differences as regards survival and the presence of metastases in patients enrolled 10 years before. In conclusion, among terminally ill cancer patients referred to a palliative home care service in Milan, mostly treated at the National Cancer Institute, the 10-year admission trend showed that palliative care is made available increasingly later, particularly to those up to 70 years of age, in contrast to current recommendations. PMID- 12088251 TI - Palliative treatment of malignant refractory ascites by positioning of Denver peritoneovenous shunt. AB - Malignant ascites is relatively common in patients with certain types of end stage cancer. Traditional treatments based on fluid and salt restriction and diuretic therapy often are not able to contain neoplastic ascites. These patients consequently undergo repeated abdominal paracentesis, with further plasma protein loss and risk of injury to abdominal viscera. The aim of this study was to evaluate our experience with Denver peritoneovenous shunt and the outcome of patients with malignant ascites and suggest some modifications to improve device patency. From February 1997 to December 1999, 44 Denver peritoneovenous shunts were placed in 42 patients, 17 women and 25 men, aged between 38 and 77 years (mean, 62.3), affected with malignant ascites due to advanced abdominal cancer. At the time of admission, 72% of patients had pain, 88% dysphagia, 66% nausea and/or vomiting, and 83% dyspnea. Eleven patients underwent local anesthesia with lidocaine and 33 general anesthesia with rapidly metabolized drugs. In 27 cases we used the peritoneal-internal jugular right vein surgical approach and in 3 cases the peritoneal-femoral vein surgical access, joining the saphena vein to the cross. In 10 cases, a radiological positioning of the Denver peritoneovenous shunt was effected by a trans-subclavian access. Relief of ascites symptoms was obtained in 87.5% of cases, with reduction of dyspnea, an increased appetite and improved ambulation. Denver peritoneovenous shunt is a good device to relieve malignant ascites, thereby reducing the risk of complications and the number of hospital admissions due to repeated paracentesis and consequently improving the quality of life. A careful patient selection, an accurate follow-up and some device modifications could improve the shunt performance, allowing a wider application of the Denver peritoneovenous shunt. PMID- 12088252 TI - Acupuncture in the treatment of menopause-related symptoms in women taking tamoxifen. AB - Fifteen patients were enrolled in a pilot study to evaluate the safety and efficacy of acupuncture for the treatment of menopausal symptoms in tamoxifen treated patients. Patients were evaluated before treatment and after one, three and six months with the Greene Menopause Index and were treated according to the traditional Chinese medicine. Anxiety, depression, somatic and vasomotor symptoms were improved by the treatment; libido was not modified. Acupuncture seems to be safe and effective for the treatment of menopausal symptoms in women with previous breast cancer taking tamoxifen. Confirmatory studies with a larger number of patients and with a placebo-treated group are warranted. PMID- 12088253 TI - Analysis of the recourse to conservative surgery in the treatment of breast tumors. AB - AIMS AND BACKGROUND: Conservative surgery is the treatment of choice for malignant tumors, at least up to stage II. The aim of this study was to analyze the recourse to conservative surgery for breast tumors and its determinants (ie, characteristics of hospitals and patients). METHODS: The study was conducted in Italy's Lazio region and was based on administrative data of the regional Hospital Information System, a database on hospitalizations. We selected all regional hospitalizations for therapeutic breast surgery over 1997, classifying them as either "conservative" or "non-conservative". The other variables considered were type of hospital, number of beds, volume of activity (average annual number of hospitalizations for breast cancer surgery), specific diagnosis, severity of cancer, and patient's age, place of residence, and socioeconomic level. A logistic model was used for multivariate analysis. RESULTS: A total of 7235 hospitalizations were analyzed, 3570 (49%) for malignant tumors and 3665 (51%) for benign disease. The logistic model showed that the factors most closely correlated with conservative surgery were age (OR = 2.2; 95% Cl: 1.8-2.6, for the age group <50 years compared to >70 years); severity of cancer (OR = 0.6; 95% Cl: 0.5-0.8, for non-localized compared to localized tumors), and volume of activity of the hospital (OR = 1.3; 95% CI: 1.0-1.6, for hospitals with >70 operations/year compared to those with <20 operations/year). The study also revealed that surgery for malignant tumors was performed by both high-volume and low-volume hospitals throughout the region. CONCLUSIONS: The association between conservative surgery and younger age, even after controlling for the severity of cancer, points to the need to encourage adherence to the existing guidelines. The association between conservative surgery and high-volume hospitals and the finding that a high proportion of breast operations is performed in low-volume facilities suggest that further efforts should be made to promote admission to high-volume hospitals. PMID- 12088254 TI - The role of brachytherapy in the management of oropharyngeal carcinomas: the Trento experience. AB - AIMS: This study was undertaken to determine the outcome of patients with oropharyngeal cancer treated at the Radiotherapy Department of the Santa Chiara Hospital (Trento, Italy) with brachytherapy alone or combined with external beam radiotherapy (EBRT). MATERIAL AND METHODS: We retrospectively reviewed the medical records of 87 patients with squamous cell carcinoma of the oropharynx treated by radiation therapy between January 1986 and September 1999. The median age was 59 years and all patients had a minimum follow-up of one year. Tumor locations were 46 tonsillar region, 31 soft palate and 10 base of the tongue. The patients were staged as follows: 41 T1, 35 T2, 11 T3 with 70 N0, 9 N1 and 8 N2. They received either brachytherapy alone (14 patients) or a combination of external beam irradiation and brachytherapy (73 patients) using an afterloading iridium technique in a plastic tube. RESULTS: Overall primary tumor control, including salvage surgery, was 81/87 (93%). Control of metastatic cervical adenopathy was as follows: clinical stage N1, 5/9 patients; N2, 2/8 patients. The estimated five-year cause-specific survival and overall survival rates were 81% and 47%, respectively. After interstitial irradiation severe complications were limited to one case of osteoradionecrosis of the mandible and seven cases of mucosal ulcer. CONCLUSION: This study confirms that iridium-192 interstitial implant alone or as a boost after external beam irradiation is a safe and effective therapy in the management of oropharyngeal carcinomas. PMID- 12088255 TI - Tumor cytosol carcinoembryonic antigen as prognostic parameter in non-small cell lung cancer. AB - AIMS AND BACKGROUND: Carcinoembryonic antigen (CEA) belongs to a family of cell surface glycoproteins. Its level in serum has a significant value for the follow up and treatment of patients with malignancies. The aim of this study was to correlate the concentration of tumor cytosol CEA (cCEA) with tumor size, patient age and sex, clinical stage, lymph node metastases, and overall survival rate in primary non-small cell lung carcinoma (NSCLC). METHODS AND STUDY DESIGN: The cCEA levels were determined in 76 NSCLC patients by luminescence assay (LIA) and radioimmunoassay (RIA). RESULTS: A strong correlation between LIA and RIA assay results was found (r = 0.992). No correlation was observed between serum CEA and cCEA levels. Tumors smaller than 3 cm had significantly higher cCEA levels than larger tumors, but when a logistic modeling process was applied this difference was not significant (P = 0.038). Histologically well-differentiated tumors also showed a significantly higher expression of cCEA (P <0.05). In addition, patients without lymph node involvement had higher cCEA levels than patients with tumor positive lymph nodes (P < 0.05). Univariate statistical analysis revealed that the risk of lymph node metastases was 1.8-fold higher in patients with low cCEA levels than in patients with higher levels, taking the median value as cutoff (P = 0.04, Kruskal-Wallis test). CONCLUSIONS: According to the results of our study, patients with overexpression of cCEA may have a better prognosis than those with low cCEA expression. cCEA might therefore be considered a good prognostic parameter as well as a prognostic factor independent of the traditional parameters for lymph node metastases. PMID- 12088256 TI - Low serum leptin level in colon cancer patients without significant weight loss. AB - AIMS AND BACKGROUND: Leptin is a protein that affects the metabolic, neuroendocrine, reproductive and hematopoietic systems and is involved in the regulation of body weight. The possible role of leptin in cancer patients, whose aforementioned systems show disorders at various levels, has been investigated by only a few studies and the results are quite contradictory. METHODS: In this study serum leptin levels were investigated in 36 patients with colon cancer having no weight loss or anorexia and in 36 healthy volunteers. Serum leptin levels were measured by radioimmunoassay. RESULTS: Significantly positive correlations were found between serum leptin level and/or body mass index (BMI) in patient and control groups (r = 0.842, P < 0.001 and r = 0.785, P < 0.001; r = 0.880, P < 0.001 and r = 0.523, P = 0.001). Serum leptin levels of colon cancer patients were significantly lower than those of the control group (8.79 vs 15.95 ng/mL, P = 0.003). BMI and age of the colon cancer patients were not different from those of the control group. Serum leptin levels of early-stage patients (n = 15) did not differ from those of advanced-stage patients (n = 21) (7.74 vs 9.54 ng/mL, P = 0.542), nor was there any difference in the serum leptin levels of patients who did and patients who did not receive chemotherapy. There was no correlation in cancer patients between serum leptin levels and CEA or CA19-9 (r = 0.015, P = 0.929 and r = 0.097, P = 0.574). CONCLUSION: Low serum leptin levels found in colon cancer patients without weight loss suggest that another mechanism regulating the leptin levels might be responsible. PMID- 12088257 TI - Erythropoietin and G-CSF receptors in human tumor cells: expression and aspects regarding functionality. AB - AIMS AND BACKGROUND: Recombinant human erythropoietin (Epo) and granulocyte colony-stimulating factor (G-CSF) are used to stimulate hematopoiesis in patients with malignant diseases. These cytokines transduce their biological signal via the Epo receptor (EpoR) and G-CSF receptor (G-CSF-R) into the cell. We therefore investigated in human tumor cell lines the expression of these receptors in tumor cells as well as their response to Epo and G-CSF. METHODS AND STUDY DESIGN: The expression of EpoR and G-CSF-R mRNA was analyzed with reverse transcription polymerase chain reaction (RT-PCR). EpoR protein expression was further monitored with Western blot and immunocytochemistry analysis. The cellular response to various concentrations of Epo was evaluated using 3[H]-thymidine uptake, Northern blot of c-fos expression and tyrosine kinase activity assay. The proliferation after G-CSF incubation was analyzed with the MTS assay. RESULTS: In this study EpoR mRNA and protein were detected in various human tumor cell lines. Treatment with Epo did not influence the proliferation rate of examined EpoR-positive tumor cell lines. Epo did not stimulate the tyrosine kinase activity nor did it affect the c-fos mRNA in these cell lines. G-CSF-R mRNA was only detected in two myeloid cell lines. Treatment with G-CSF did not increase the proliferation of these cells. CONCLUSIONS: These results demonstrate that Epo and G-CSF did not modulate the growth rate of examined receptor-positive tumor cell lines; the presence of the Epo receptor seems not essential for cell growth of these tumor cells in cell culture. PMID- 12088258 TI - Osteosarcoma of the sternum: two case reports. AB - Primary sarcomas of the sternum are infrequent; the most common histological types are chondrosarcoma, plasmacytoma, Ewing's sarcoma and Hodgkin's tumors. Since osteosarcoma of the sternum is a very rare entity, we have thought to explain the principal characteristics and the clinical and therapeutic approach based on our experience of two cases. PMID- 12088259 TI - Gastric adenocarcinoma associated with granulomatous gastritis. Case report and review of the literature. AB - AIMS: We describe the fourth reported case of granulomatous gastritis associated with gastric adenocarcinoma, with a review of the literature and considerations about the prognostic implications of this association. RESULTS: A 48-year-old woman who had been suffering from gastritis for ten years was admitted to our institute for increasing left epigastric pain associated with vomiting. After an endoscopic biopsy had revealed an ulcerated signet ring cell carcinoma, the patient was submitted to subtotal gastrectomy with regional lymph node dissection. Pathological examination of the resected specimen revealed a superficial signet ring cell carcinoma (early cancer) associated with multiple granulomas. The granulomas, which were observed within the mucosa and the submucosa at the periphery of the carcinoma, were composed of CD68-positive, CD15 negative epithelioid and giant cells of the Langhans type, confirming their true histiocytic nature, and were also extensively found within the dissected lymph nodes. Since no ocular, skin, pulmonary or other gastrointestinal lesions were found and the granulomas were negative for acid-fast and fungal stain, a diagnosis of granulomatous gastritis was made. CONCLUSIONS: To the best of our knowledge this is the fourth example of gastric adenocarcinoma and granulomatous gastritis. These cases suggest an association between granulomatous gastritis and early gastric cancer. PMID- 12088261 TI - Squamous cell vocal cord carcinoma and synchronous small cell lung carcinoma: a case report. AB - According to various authors the incidence of multiple primary tumors in the aerodigestive tract can be considered high. However, most of these tumors are metachronous and of the same histological type. The occurrence of a small cell lung carcinoma synchronous with a squamous cell vocal cord carcinoma is a very rare phenomenon. In this report we present the second case in the English literature. PMID- 12088260 TI - Long-term survival after regional chemotherapy for liver metastases from breast cancer. A case report. AB - The prognosis of patients with liver metastases from breast cancer has to be regarded as rather unfavorable. A 45-year-old woman with cancer of the left breast and multiple simultaneous liver metastases was initially treated with cyclophosphamide, methotrexate, and 5-fluorouracil. After two treatment cycles a sonogram of the left breast revealed reduction of the tumor size and she underwent a modified radical mastectomy and hepatic artery catheterization. There was no change in the multiple lesions of the liver on abdominal ultrasonography. One week later a regional chemotherapy regimen was given through a hepatic arterial catheter with subcutaneous implanted reservoir. After completion of the seventh course of chemotherapy, ultrasonography revealed that the multiple liver metastases had regressed completely. To date (July 2001) there has been no relapse during the seven-year follow-up period. In conclusion, we suggest that intra-arterial regional chemotherapy may have an important role to play in the management of breast cancer patients with isolated liver metastases. PMID- 12088262 TI - Pulmonary blastoma after liver transplant: a case report. AB - There is an increased risk of cancer after organ transplantation mainly due to the immunosuppressive therapy required in these patients. We report a case of biphasic pulmonary blastoma in an adult male who underwent liver transplant for hepatocellular carcinoma in March 1999, followed by immunosuppressive treatment and adjuvant chemotherapy with epirubicin. Disease-free survival lasted 18 months, then a diagnosis of biphasic pulmonary blastoma was made and the patient underwent a lung lobectomy. Five months after surgical resection a recurrence of this rare tumor was recorded and two cycles of cisplatin + etoposide and ifosfamide + etoposide and one cycle of second-line chemotherapy with vinorelbine were administered. The tolerability and the efficacy of this treatment were poor. The patient died less than one year after diagnosis. To our knowledge this is the first reported case of pulmonary blastoma in a transplant patient. Our findings confirm that organ transplant recipients deserve long-term medical surveillance also in the absence of graft complications, and that pulmonary blastoma is an aggressive tumor with a poor prognosis. PMID- 12088263 TI - Independent clinical trials: a commentary. AB - The so-called norms of good clinical practice have been incorporated into the Italian regulatory legislation governing clinical trials sponsored by pharmaceutical companies, but there are no legislative provisions governing independent clinical trials: ie those not sponsored by the industry. The pharmaceutical industry has recently increased considerably its commitment to sponsored trials by establishing a series of economic relationships with individual researchers and hospital or university institutions. It has also set up and strengthened a series of bodies and service companies with the aim of making the clinical trials "machine" more efficient. Such developments have aroused alarm in the medical literature because of the risk that they may have negative effects on the freedom of research and research results. At the same time, there is also the risk that independent clinical trials will be greatly penalized by having to compete with sponsored trials in terms of patient enrollment, and because they are currently having to face a series of difficulties connected with the lack or scarcity of economic resources provided by the State or non-profit organizations, with problems relating to patient insurance and to the availability of the necessary drugs. However, the objective of independent trials is to improve the medical art by answering specific diagnostic and therapeutic questions, whereas that of industry-sponsored trials is to generate money, directly or indirectly, by means of the registration of new drugs. It is therefore now necessary to ensure better surveillance of the influence of pharmaceutical companies over the trials they sponsor (as a minimum by ensuring the transparency of a series of potential conflicts of interest between them and clinical researchers) and, simultaneously, protect independent trials from coming to an inglorious end by means of specific support initiatives such as those proposed in this article. PMID- 12088264 TI - Cancer mortality in Italy, 1998. AB - This short report provides data and statistics of cancer mortality in Italy in 1998, updating previous work on the issue. The material and methods of this report are similar to those previously described. Briefly, cancer death certification numbers by cause and estimates of the resident population in 1998, stratified by sex and quinquennia of age, were abstracted from data provided by the Istituto Nazionale di Statistica (ISTAT). All cancers or groups of cancers, classified according to the standard International Classification of Diseases (ICD), Ninth Revision, were grouped in 31 categories, besides total cancer mortality and other and unspecified sites. We grouped together all intestinal sites, melanomas and non-melanomatous skin neoplasms, all uterine neoplasms (cervix and corpus), all neoplasms of the brain and nerves (benign and malignant), all leukemias, and all non-Hodgkin's lymphomas. Eight tables were produced, including the following statistics: 1) number of deaths, crude and age standardized death certification rates, and percentages of all cancer deaths for population at all ages and truncated 35-64 years (Table 1 for males and Table 2 for females). Two different standards were used: i) the 1971 Italian census population, corrected for census undercount and subdivided into 16 quinquennia of age from 0-4 to 75-79, plus 80 and over, and ii) the world standard population, for purposes of comparison with other countries; 2) age-specific death certification rates for each sex and quinquennium of age from 0-4 to 75-79, plus 80 and over (Table 3 for males and Table 4 for females); 3) total number of registered deaths for each cancer or group of cancers, sex and age group (Table 5 for males and Table 6 for females); 4) percentage of all cancer deaths for each sex and age group (Table 7 for males and Table 8 for females). A few comments are included, mainly in order to assist reading and interpretation of data for major cancer sites, and to recall underlying long-term tendencies. Any inference should in any case be based on age-standardized rates, and, essentially, on detailed inspection of age-specific rates. PMID- 12088265 TI - Role of double-contrast barium enema in colorectal cancer screening based on fecal occult blood. AB - AIMS AND BACKGROUND: Screening for colorectal cancer by fecal occult-blood testing has been shown to be effective in reducing colorectal cancer mortality. Total colonoscopy is the test of choice for the assessment of fecal occult blood positive subjects. Double-contrast barium enema is commonly employed to study the rest of the colon when colonoscopy is incomplete. The present study evaluated the contribution of double-contrast barium enema in detecting neoplastic lesions of the colon in fecal occult-blood-positive subjects with incomplete colonoscopy. METHODS: In the frame of a screening program for colorectal cancer in the Florence District, a new immunochemical fecal occult-blood test replaced the classic guaiac fecal occult-blood test in 1993. Subjects with a positive fecal occult-blood test were invited to undergo total colonoscopy. Incomplete colonoscopy prompted double-contrast barium enema. Type and rate of neoplastic lesions detected by endoscopy or double-contrast barium enema as single methods or combined were evaluated. RESULTS: A total of 38,829 subjects underwent fecal occult-blood testing in the period 1993-2000. Overall, 1,542 were positive. Assessment was refused by 235 subjects. Out of 1,307 subjects accepting assessment, total colonoscopy was attempted in 1,294: of these, it was not possible in 343 cases, and double-contrast barium enema was advised and performed in 261 subjects. Colorectal cancer was detected in 115 subjects, single or multiple adenomas in 323, hyperplastic polyps in 58, inflammatory, hamartomatous or not histologically confirmed polyps in 38, and other benign non-polypoid findings or no abnormality in 773. There were significant differences between the rates of detected colorectal cancers or adenomas of total and incomplete colonoscopy. There were also significant differences between incomplete colonoscopy and the combination of incomplete colonoscopy and double-contrast barium enema as regards rates of detected colorectal cancer, and between total colonoscopy and the combination of incomplete colonoscopy with double-contrast barium enema as regards rates of detected adenomas. Double-contrast barium enema associated to incomplete colonoscopy was responsible for an increase in detection rates of cancer or adenoma of 2.3/1000 or 3.8/1000, respectively. CONCLUSIONS: Double-contrast barium enema was useful in detecting colorectal cancer beyond the range reached by incomplete colonoscopy, whereas our data confirmed a lower sensitivity of double-contrast barium enema for polyps. The diagnostic contribution observed in the present survey confirms the opportunity of performing double-contrast barium enema as a routine adjunct to incomplete colonoscopy. Nevertheless, in order to maximize the detection rate of adenomas, the rate of total colonoscopy should be kept as high as possible. PMID- 12088266 TI - Docetaxel followed by gemcitabine in the treatment of advanced non-small cell lung cancer: a phase I study. AB - AIMS AND BACKGROUND: Based on the results of a preclinical study, a phase I trial was conducted to evaluate the feasibility of administering docetaxel followed by gemcitabine in non-small cell lung cancer patients. STUDY DESIGN: Sixteen patients with advanced non-small cell lung cancer (stages III B-IV) were treated on the 1st day with docetaxel and on the 8th day with gemcitabine. Treatment was repeated every three weeks for a maximum of six cycles. Five groups received docetaxel/gemcitabine (mg/ml): 50/800, 60/800, 60/900, 60/1,000, 70/1,000. All patients and 57 cycles were assessed for toxicity. RESULTS: The most important side effects were grade IV neutropenia in 4 patients (2 at the 60/1000 level and 2 at the 70/1000 level) and grade III leukopenia and neutropenia without fever in 4 and 6 patients, respectively. Maximum tolerated dose was not reached. CONCLUSIONS: The sequence docetaxel-gemcitabine appears well tolerated and easy to administer. For this reason, a phase II study is ongoing to fully assess its antitumor activity. PMID- 12088267 TI - Asthma therapy in the new millennium. AB - Bronchial asthma is one of the most common chronic diseases in modern society and yet, despite the availability of highly effective drugs, there is increasing evidence to suggest that its incidence is increasing. It is a general health problem in several industrialised countries and will remain one for the next decades. With regard to asthma pathogenesis, our understanding has increased tremendously over the last two decades. Therefore, the potential for specific targeted and constructed therapies has become evident. Monoclonal antibodies to IgE, soluble receptors or antibodies to certain cytokines such as IL-4 and IL-5 are being investigated as possible treatments for asthma. Besides the already known receptor antagonists, new compounds directed to novel receptor types (e.g. cytokine, adenosine, adhesion molecules, etc.) are now under development. New targets in the cytosol will come into focus. Preliminary studies of selective phosphodiesterase (PDE) inhibitors in asthmatic patients have been encouraging. It is also very likely that the use of glucocorticoids cannot be excluded from therapy. However, we should generate new glucocorticoids with less side-effects, probably by using the so-called retrometabolic drug design. The first representative of this new steroid class, loteprednol is already approved for the therapy of certain allergic disorders. Because asthma is a disease of many different gene polymorphisms, gene therapy seems to be of low success at present. Alternatively, antisense oligonucleotides could be used. Future developments may also include strategies targeting the remodeling of structural elements of the airways. Today's intensive search for new treatments should ensure a greater diversity of therapeutic possibilities for the management of asthma in the next millennium. PMID- 12088268 TI - Tea tree oil reduces histamine-induced oedema in murine ears. AB - OBJECTIVE: To examine the effect of topically applied tea tree oil (TTO) on histamine-induced oedema in the ears of mice. METHODS AND RESULTS: For BALB/c mice, 10 microl undiluted TTO applied immediately after, but not 30 min before intradermal injection of 600 microg histamine in 10 microl, significantly suppressed oedema development. TTO applied after histamine injection also suppressed histamine-induced oedema in C57/BL6 mice. TTO applied immediately after intradermal injection of compound 48/80 (200 microg in 10 microl saline) also significantly reduced ear swelling. TTO suppressed histamine-induced oedema to the same extent in capsaicin-treated (neuropeptide-depleted) and control mice which suggests that TTO does not inhibit histamine-induced oedema by regulating the activity of peripheral sensory neurons. Terpinen-4-ol, the major water soluble component of TTO, was equivalent in potency to TTO in the suppression of histamine-induced ear swelling. CONCLUSION: Topical application of TTO, and in particular terpinen-4-ol, may be effective in controlling histamine-induced oedema often associated with Type I allergic immediate hypersensitivities. PMID- 12088269 TI - Soluble HLA-II expression in African-Americans. AB - OBJECTIVE: In this study we evaluated the contribution of major histocompatibility complex (MHC) genes to soluble histocompatibility antigen class II (sHLA-II) secretion in African American patients with rheumatoid arthritis (RA). METHODS: A sensitive enzyme-linked immunoassay was used to quantitate sHLA-II in the serum of 7 patients with RA, as well as 28 of their kinships and 49 HLA typed normal African American individuals. RESULTS: Mean sHLA II values were higher in patients with RA than those in healthy African American individuals (p < 0.05). There were variations in concentrations in individual patients but these were unrelated to any apparent clinical event. The proportion of unaffected family members with detectable levels of sHLA-II was not significantly different than those in normal controls. Neither specific HLA haplotype, or HLA-allele(s) correlated with high or low sHLA-II secretion. CONCLUSIONS: Our data suggest that sHLA-II molecules are not regulated by MHC linked genes but may be regulated by non-MHC linked genes and racial background may reflect genetic heterogeneity of the expression of this soluble HLA material. These observations contrast with previous observations concerning soluble HLA class I (sHLA-I) molecules in a described population sample which were almost the precise reverse. PMID- 12088270 TI - B1 and B2 antagonists and bradykinin-induced blood flow in rat skin inflammation. AB - OBJECTIVE AND DESIGN: The aim of the present study was to examine the contribution of the two kinin receptors B1 and B2 to the increased blood flow observed in response to bradykinin (BK) in a blister model under different injury conditions. MATERIAL: Young male Sprague-Dawley rats weighing 250-350 g were used. METHODS: A vacuum-induced blister was raised in the rat hind paw and blood flow measured in the superfused blister base under four different conditions including, early phase acute injury; late phase acute injury; recurrent injury and early phase acute injury in the setting of chronic nerve damage. BK (10 microM) was superfused alone, or in the presence of the B1 antagonist DesArg9Leu8BK (DALBK), (10 nM) and/or the B2 antagonist [D-Arg,Hyp3,Thi5 D Tic7,Oic8] Bradykinin (HOE 140) (10 nM). RESULTS: HOE 140 significantly inhibited the BK response in all models. Significant inhibition of BK-induced vasodilatation by DALBK was only observed in the late phase acute and recurrent injury models. CONCLUSIONS: The results suggest that the involvement of the inducible B1 receptor in skin inflammation site is related to the site, duration and recurrence of the injury condition. PMID- 12088271 TI - Time-dependent expression of annexin 1 in a model of chronic granulomatous inflammation. AB - OBJECTIVE AND DESIGN: To determine the expression pattern and distribution of the glucocorticoid-inducible protein annexin 1 (ANXA1) in a murine model of chronic granulomatous inflammation. MATERIALS OR SUBJECTS: TO Mouse. TREATMENT: Chronic granulomatous inflammation was induced by injecting into dorsal sub-cutaneous air pouches in mice, a mixture of croton oil and Freund's complete adjuvant (CO/FCA). METHODS: Western and northern analysis, corticosterone assay, and immunohistochemistry. Statistical analysis was performed using ANOVA followed by Tukey's pair-wise comparisons or Dunnett's multiple comparisons. RESULTS: ANXA1 protein levels changed significantly throughout the 4-week time course, with an initial peak at day 7 and a later elevation at 28 days. ANXA1 mRNA levels peaked at days 1 and 3, with a significant decline at day 7 followed by an upward trend to day 28. Plasma corticosterone measurements taken throughout the time course revealed an increase from 14 days onward, suggesting that corticosterone does not influence ANXA1 expression during the initial stages of the model. Immunogold staining revealed that ANXA1 expression in the inflamed tissue was mainly in extravasated neutrophils, with intact protein (37 kDa) being predominantly observed on the cell membrane. CONCLUSIONS: The pattern of ANXA1 expression indicates that infiltrated neutrophils are responsible for the majority of ANXA1 present both at early and later stages of this model of granulomatous inflammation. PMID- 12088272 TI - Time course study on the development of allergen-induced airway remodeling in mice: the effect of allergen avoidance on established airway remodeling. AB - OBJECTIVE AND DESIGN: We carried out a time course study on the development of allergen-induced airway remodeling in a mouse model of allergic asthma. Moreover, we examined the effect of allergen avoidance on the established airway remodeling. MATERIALS AND METHODS: BALB/c mice were sensitized to ovalbumin (OA) with alum, and exposed daily for 3 weeks to aerosolized OA. At each designated point, bronchial responsiveness was measured, and bronchoalveolar lavage and histological examination were carried out. RESULTS: The numbers of inflammatory leukocytes in the airways and the percentage of goblet cells in the epithelium, Th2 cytokine production, IgE production, collagen deposition beneath the basement membrane and bronchial responsiveness to acetylcholine were all markedly increased after repeated antigen challenge for 1-3 weeks. In contrast, after cessation of antigen exposure, goblet cell hyperplasia, inflammatory infiltrates and bronchial hyperresponsiveness were gradually attenuated and had almost resolved 4 weeks after cessation, but subepithelial fibrosis was still observed at this time point. CONCLUSIONS: The present findings demonstrated that epithelial changes following repeated allergen challenge are rapidly induced and recover after the cessation of exposure, but subepithelial fibrosis has a late onset and relatively irreversible changes, and subepithelial fibrosis in contrast to goblet cells hyperplasia did not appear to contribute to bronchial hyperresponsiveness, at least, in this mouse model. PMID- 12088274 TI - Polyisoprenyl phosphates: natural antiinflammatory lipid signals. AB - Lipoxins (LX) and aspirin-triggered 15-epimer LX are leukocyte-derived eicosanoids generated during host defense that serve as down-regulatory signals. The specific intracellular events that govern cellular responses to inhibitory extracellular signals are of wide interest in order to understand pivotal intracellular events in diseases characterized by enhanced inflammatory responses, such as asthma, rheumatoid arthritis and atherosclerosis. We recently uncovered a novel role for polyisoprenyl phosphates, in particular presqualene diphosphate (PSDP), as natural down-regulatory signals in human neutrophils that directly inhibit phospholipase D and superoxide anion generation. Activation of LXA4 receptors (ALXR) reverses proinflammatory receptor-initiated decrements in PSDP and inhibits cellular responses. These findings represent evidence for a novel paradigm for lipid-protein interactions in the control of cellular responses, namely receptor-initiated degradation of repressor lipids that is subject to regulation by aspirin treatment via the actions of aspirin-triggered 15-epimer LX at the ALXR, and identify new templates for antiinflammatory drugs by design. PMID- 12088275 TI - Enzymes and receptors in the leukotriene cascade. AB - Leukotrienes are a family of paracrine hormones derived from the oxidative metabolism of arachidonic acid. These lipid mediators are recognized as important signal molecules in a variety of inflammatory and allergic conditions affecting the skin, joints, gastrointestinal and respiratory systems, in particular asthma. Such conditions are typified by local pain, tissue edema, hyperemia and functional losses. In the tissues, immunocompetent cells accumulate at the site of injury which contribute to tissue damage and perpetuation of the disease process. Leukotrienes can elicit most, if not all, of these signs and symptoms. Thus, leukotriene B4 is one of the most powerful chemotactic agents known to date and participates in the recruitment of leukocytes. The cysteinyl leukotrienes, on the other hand, contract smooth muscles, particularly in the peripheral airways and microcirculation. Recently, drugs which block the formation and action of leukotrienes have been introduced as novel antiasthmatic medications. This chapter reviews the biochemistry, molecular biology and cell biology of the key enzymes and cognate receptors in the leukotriene cascade. PMID- 12088277 TI - Phosphoinositides and signal transduction. AB - Phosphoinositides comprise a family of eight minor membrane lipids which play important roles in many signal transducing pathways in the cell. Signaling through various phosphoinositides has been shown to mediate cell growth and proliferation, apoptosis, cytoskeletal changes, insulin action and vesicle trafficking. A number of advances in signal transduction in the last decade has resulted in the discovery of a growing list of proteins which directly interact with high affinity and specificity with distinct phosphoinositides. Equally important, a number of phosphoinositide binding domains such as the pleckstrin homology domain have emerged as critical mediators of phosphoinositide signaling. Here, recent advances in phosphoinositide signaling are discussed. The aim of this review is to highlight particularly exciting advances made in the field over the last few years. The regulation of phosphoinositide metabolism by lipid kinases, phosphatases and phospholipases is reviewed, and considerable emphasis is placed on phosphoinositide-binding proteins. Finally, the role of these lipids in regulating signaling pathways and cell function is described. PMID- 12088276 TI - Lipid mediators in epithelial cell-cell interactions. AB - Epithelial cells which line mucosal surfaces (e.g. lung, intestine) play a central role in the coordination of the inflammatory response. In both the healthy and diseased mucosa, epithelia lie anatomically positioned in close proximity to a number of other cell types, including leukocytes, fibroblasts, smooth muscle cells and vascular endothelia. This complex architecture supports a unique microenvironment for biochemical cell-cell crosstalk. Our previous studies and work by others have elucidated lipid mediator signaling networks emanating from epithelial cell-cell interactive pathways, and have defined a number of targets for development of effective therapeutics. This short review will focus on recently defined pathways of lipid mediator function in the mucosa, particularly with regard to the role of the epithelium. PMID- 12088278 TI - Microsomal cytochrome P450 and eicosanoid metabolism. AB - The demonstration of a role for microsomal P450 in the metabolism of endogenous pools of arachidonic acid established this enzyme system as a member of the arachidonic acid cascade and characterized a new an important metabolic function for this enzyme system. Studies from several laboratories documenting the powerful biological activities of the P450-derived eicosanoids have suggested important roles for the P450 arachidonic acid monooxygenase in renal and vascular physiology, and in the pathophysiology of experimental hypertension. These studies provide significant evidence to indicate that in addition to its recognized traditional toxicological and pharmacological roles, microsomal P450s also play important physiological roles in the control of tissue and body homeostasis. PMID- 12088279 TI - PPARs: transcriptional effectors of fatty acids and their derivatives. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear hormone receptors that mediate the effects of fatty acids and their derivatives at the transcriptional level. These receptors stimulate transcription after activation by their cognate ligand and binding to the promoter of target genes. In this review, we discuss how fatty acids affect PPAR functions in the cell. We first describe the structural features of the ligand binding domains of PPARs, as defined by crystallographic analyses. We then present the ligand-binding characteristics of each of the three PPARs (alpha, beta/delta, gamma) and relate ligand activation to various cellular processes: (i) fatty acid catabolism and modulation of the inflammatory response for PPARalpha, (ii) embryo implantation, cell proliferation and apoptosis for PPARbeta, and (iii) adipocytic differentiation, monocytic differentiation and cell cycle withdrawal for PPARgamma. Finally, we present possible cross-talk between the PPAR pathway and different endocrine routes within the cell, including the thyroid hormone and retinoid pathways. PMID- 12088280 TI - Cyclooxygenase, lipoxygenase and tumor angiogenesis. AB - Arachidonic acid metabolism through cyclooxygenase (COX) and lipoxygenase (LOX) pathways generates various biologically active lipids that play important roles in inflammation, thrombosis and tumor progression. Angiogenesis, the formation of new capillary vessels from preexisting ones, underpins a number of physiological processes and participates in the development of several pathological conditions such as arthritis, cancer and various eye diseases. The formation of new capillary vessels is a multistep process that involves endothelial cell proliferation, migration and tube formation. In the present review, we survey the literature on the regulation of angiogenesis by arachidonate metabolites, especially those from the COX and 12-LOX pathways in the context of tumor growth, and put forward some unanswered but important questions for future studies. PMID- 12088281 TI - Products of the isoprostane pathway: unique bioactive compounds and markers of lipid peroxidation. AB - We previously reported the discovery of prostaglandin F2-like compounds (F2 isoprostanes) formed by nonenzymatic free-radical-induced peroxidation of arachidonic acid. Quantification of F2-isoprostanes has proven to be a major advance in assessing oxidative stress status in vivo. Central in the pathway of formation of isoprostanes are prostaglandin H2-like endoperoxides, which also undergo rearrangement in vivo to form E-ring, D-ring, and thromboxane-ring compounds. E2- and D2-isoprostanes also undergo dehydration in vivo to form reactive cyclopentenone A2- and J2-isoprostanes, which are susceptible to Michael addition reactions with thiols. Recently, we described the formation of highly reactive gamma-ketoaldehydes (now termed isoketals) as products of isoprostane endoperoxide rearrangement which readily adduct to lysine residues on proteins and induce cross-links at rates that far exceed other aldehyde products of lipid peroxidation. Isoprostane-like compounds (neuroprostanes) and isoketal-like compounds (neuroketals) are formed from oxidation of docosahexaenoic acid, which is enriched in the brain, and measurement of neuroprostanes may provide a unique marker of oxidative neuronal injury. PMID- 12088282 TI - Human estrogen receptor-alpha: regulation by synthesis, modification and degradation. AB - This review aims to evaluate the impact that human estrogen receptor-alpha (ER alpha) synthesis, modification and degradation has on estrogen-dependant physiological and pathological processes within the body. Estrogen signaling is transduced through estrogen receptors, which act as ligand-inducible transcription factors. The significance of different isoforms of ER-alpha that lack structural features of full-length ER-alpha are discussed. The influence of differential promoter usage on the amount and isoform of ER-alpha within individual cell types is also reviewed. Moreover, the potential role of phosphorylation, ubiquitination and acetylation in the function and dynamic turnover of ER-alpha is presented. PMID- 12088283 TI - Pore-forming toxins. AB - Pore-forming toxins are widely distributed proteins which form lesions in biological membranes. In this review, bacterial pore-forming toxins are treated as a paradigm and discussed in terms of the structural principles on which they work. Then, a large family of bacterial toxins, the cholesterol-binding toxins, are analyzed in depth to provide an overview of the processes involved in pore formation. The ways in which the cholesterol-binding toxins (cholesterol dependent cytolysins) interact with membranes and form pores, the structure of the monomeric soluble and oligomeric pore-forming states, and the effects of the toxin on membrane structure are discussed. By surveying the range of work which has been done on cholesterol-binding toxins, a working model is elaborated which reconciles two current, apparently diametrically opposed, models for their mechanism. PMID- 12088284 TI - Sleeping sickness and the brain. AB - Recent progress in understanding the neuropathological mechanisms of sleeping sickness reveals a complex relationship between the trypanosome parasite that causes this disease and the host nervous system. The pathology of late-stage sleeping sickness, in which the central nervous system is involved, is complicated and is associated with disturbances in the circadian rhythm of sleep. The blood-brain barrier, which separates circulating blood from the central nervous system, regulates the flow of materials to and from the brain. During the course of disease, the integrity of the blood-brain barrier is compromised. Dysfunction of the nervous system may be exacerbated by factors of trypanosomal origin or by host responses to parasites. Microscopic examination of cerebrospinal fluid remains the best way to confirm late-stage sleeping sickness, but this necessitates a risky lumbar puncture. Most drugs, including many trypanocides, do not cross the blood-brain barrier efficiently. Improved diagnostic and therapeutic approaches are thus urgently required. The latter might benefit from approaches which manipulate the blood-brain barrier to enhance permeability or to limit drug efflux. This review summarizes our current understanding of the neurological aspects of sleeping sickness, and envisages new research into blood-brain barrier models that are necessary to understand the interactions between trypanosomes and drugs active against them within the host nervous system. PMID- 12088285 TI - Carbohydrates as templates for control of distance-geometry in de novo-designed proteins. AB - An understanding of very complex natural systems can often only be achieved through detailed studies of systems with a reduced complexity. Thus, de novo design of proteins allows the study of fundamental forces determining protein folding and stability, as well as protein-protein interactions, by analyses of protein models of structural motifs. In addition, de novo design may lead to new biomimetic molecules with novel properties. In a synthetic approach to achieve structural economy, rigid templates, sometimes called topological scaffolds, have been used to connect secondary-structure elements, most notably alpha-helices. By positioning the helices on the template, the unfavorable entropy of protein folding is reduced. In a novel class of chimeric molecules called carboproteins, carbohydrates are used as templates for de novo design of protein models. Recently, a strategy relying on chemoselective ligation of C-terminal peptide aldehydes to tetra-aminooxy functionalized monosaccharides has provided 7-kDa 4 alpha-helix bundle carboproteins. PMID- 12088286 TI - P2X4 and P2X6 receptors associate with VE-cadherin in human endothelial cells. AB - We investigated the expression of P2X4 and P2X6 receptors on human umbilical vein endothelial cells (HUVECs) and found that both P2X receptor subtypes on plasma membranes are largely restricted to areas of cell-cell contact. Co-labelling experiments at the confocal and electron microscopy levels revealed that P2X4 and P2X6 receptors are strongly co-localised with the cell adhesion molecule VE cadherin. The P2X4 and P2X6 receptors on plasma membranes at cellular junctions are rapidly (within 5 min) internalised specifically after decreasing extracellular [Ca2+]. Disruption of microfilaments, microtubules and integrin mediated adhesion or stimulation of P2 receptors with ATP did not alter P2X4 and P2X6 receptor expression on HUVEC plasma membranes. Membraneous P2X4 and P2X6 receptors resisted extraction with Triton-X 100, whereas cytoplasmic P2X receptors were Triton-X 100 soluble. P2X4 receptors, but not P2X6 receptors, could be co-immunoprecipitated with VE-cadherin and vice versa. We conclude that P2X4 and P2X6 receptors are associated with VE-cadherin at HUVEC adherens junctions. PMID- 12088287 TI - Chronic gestational exposure to ethanol impairs insulin-stimulated survival and mitochondrial function in cerebellar neurons. AB - Chronic gestational exposure to ethanol has profound adverse effects on brain development. In this regard, studies using in vitro models of ethanol exposure demonstrated impaired insulin signaling mechanisms associated with increased apoptosis and reduced mitochondrial function in neuronal cells. To determine the relevance of these findings to fetal alcohol syndrome, we examined mechanisms of insulin-stimulated neuronal survival and mitochondrial function using a rat model of chronic gestational exposure to ethanol. In ethanol-exposed pups, the cerebellar hemispheres were hypoplastic and exhibited increased apoptosis. Isolated cerebellar neurons were cultured to selectively evaluate insulin responsiveness. Gestational exposure to ethanol inhibited insulin-stimulated neuronal viability, mitochondrial function, Calcein AM retention (membrane integrity), and GAPDH expression, and increased dihydrorosamine fluorescence (oxidative stress) and pro-apoptosis gene expression (p53, Fas-receptor, and Fas ligand). In addition, neuronal cultures generated from ethanol-exposed pups had reduced levels of insulin-stimulated Akt, GSK-3beta, and BAD phosphorylation, and increased levels of non-phosphorylated (activated) GSK-3beta and BAD protein expression. The aggregate results suggest that insulin-stimulated central nervous system neuronal survival mechanisms are significantly impaired by chronic gestational exposure to ethanol, and that the abnormalities in insulin signaling mechanisms persist in the early postnatal period, which is critical for brain development. PMID- 12088288 TI - Protein kinases mediate nitric oxide-induced apoptosis in the insect cell line IPLB-LdFB. AB - The involvement of protein kinases (PKA, PKC and PKB) in nitric oxide (NO) induced apoptosis with sodium nitroprusside plus N-acetyl-L-cysteine in the IPLB LdFB cell line from the insect Lymantria dispar was investigated. The presence of protein kinase-like molecules was demonstrated by Western blot analysis. The role of the kinases in programmed cell death was analysed in cytofluorimetric experiments by incubating the insect cells with H-89 (a specific inhibitor of PKA), calphostin C (an inhibitor of PKC) or wortmannin (an inhibitor of phosphatidylinositol 3-kinase). The results show that PKA is correlated with the induction and PKC and PKB with the prevention of NO-induced insect cell death. Moreover, NO-induced apoptosis involves the release of cytochrome c. PMID- 12088289 TI - Gluten-free diet and quality of life in patients with screen-detected celiac disease. AB - CONTEXT: Since the advent of serologic testing for celiac disease, most persons who receive a diagnosis of celiac disease have few or no symptoms. Although pathologic changes of celiac disease resolve on a gluten-free diet, how a gluten free diet affects the quality of life for patients with screen-detected celiac disease is unclear. OBJECTIVE: To evaluate the effect of a gluten-free diet on the quality of life of patients with screen-detected celiac disease. DESIGN: Prospective study of patients before and 1 year after initiating a gluten-free diet. PARTICIPANTS: 19 patients with screen-detected celiac disease (found by serologically testing first-degree relatives of celiac patients) and 21 consecutive patients with symptom-detected disease. In all cases, celiac diagnosis was confirmed by finding villous atrophy and crypt hyperplasia on small bowel biopsy. INTERVENTION: Gluten-free diet (explained during a single physician visit). MAIN OUTCOME MEASURES: Gastrointestinal Symptoms Rating Scale (GSRS), in which scores range from 0 to 6 (higher scores represent worse symptoms); and quality of life measured with the Psychological General Well-Being Questionnaire (PGWB). Scores range from 22 to 132 (higher scores mean greater well-being). RESULTS: At baseline, patients with symptom-detected celiac disease had poorer quality of life and more gastrointestinal symptoms than those with screen detected celiac disease. Reported compliance with the gluten-free diet was good. All mucosal lesions of the small bowel had resolved at the follow-up biopsy. After 1 year of following the diet, quality of life for patients with screen detected disease significantly improved (mean PGWB score increased from 108 to 114; P <0.01). A similar increase was noted in patients with symptom-detected disease (mean PGWB score increased from 92 to 103; P <0.01). Gastrointestinal symptoms also improved in patients with screen-detected disease and in patients with symptom-detected disease (mean GSRS scores decreased from 1.8 to 1.4 and from 2.6 to 1.9, respectively; P <0.01 for both comparisons). CONCLUSIONS: Gluten free diet was associated with improved quality of life for patients with symptom detected celiac disease and patients with screen-detected celiac disease. Concerns about the burden of a gluten-free diet, at least over the short term, may be unfounded. PMID- 12088290 TI - Use and evaluation of critical pathways in hospitals. AB - CONTEXT: Although hospitals have devoted substantial resources to critical pathways, it is not known whether they routinely evaluate the clinical or economic effects of these pathways. OBJECTIVE: To determine how use and evaluation of critical pathways differ between academic and community hospitals. DESIGN: Cross-sectional survey. PARTICIPANTS: Hospitals participating in consortia for improving quality of care associated with the Institute of Health Care Improvement and the VHA, Inc. (formerly known as the Voluntary Hospitals of America, Inc.). Hospital administrators at 41 hospitals completed the survey (71% response rate), representing 13 academic medical centers, 13 community teaching hospitals, and 15 community hospitals. MEASURES: Use of critical pathways and measurement of clinical and economic outcomes of pathways. RESULTS: The median number of adult critical pathways used by academic hospitals, community teaching hospitals, and community hospitals was 25, 18, and 3, respectively. The most common pathways were community-acquired pneumonia, total hip or knee replacement, and stroke or transient ischemic attack. The percentage of hospitals with pathways dedicating staff to manage them was 78% for academic hospitals, 22% for community teaching hospitals, and 14% for community hospitals (P = 0.02). Evaluation practices varied widely among hospitals with pathways. Measures assessed included monitoring length of stay (85%), total hospital costs (74%), in hospital mortality (62%), infectious complications (53%), readmission rates (47%), functional status (18%), and adverse drug events (15%). CONCLUSION: The use of critical pathways varies substantially among hospitals participating in quality improvement consortia. Use was highest in academic centers and lowest in community hospitals. Many hospitals with pathways do not track important clinical outcomes as part of their evaluation practices. PMID- 12088291 TI - How underlying patient beliefs can affect physician-patient communication about prostate-specific antigen testing. AB - CONTEXT: Routine cancer screening with prostate-specific antigen (PSA) is controversial, and practice guidelines recommend that men be counseled about its risks and benefits. OBJECTIVE: To evaluate the process of decision making as men react to and use information after PSA counseling. DESIGN: Written surveys and semistructured qualitative interviews before and after a neutral PSA counseling intervention. PARTICIPANTS: Men 40 to 65 years of age in southeastern Michigan were recruited until thematic saturation--that is, the point at which no new themes emerged in interviews (n = 40). RESULTS: In a paper survey, 37 of 40 participants (93%) said that they interpreted the counseled information as unfavorable toward PSA. However, 30 participants (75%) said after the intervention that they intended to be tested in the future, including 29 of 30 men (97%) with prior PSA testing. In the interview, many participants cited underlying beliefs as a reason to dismiss the counseled information. Qualitative analysis found the seven most common beliefs cited were fear of cancer, relevance of salient anecdotes and analogies, distrust of statistics, enthusiasm for "prevention," protection from "bad luck," faith in science, and valuing PSA as knowledge for its own sake. Although some beliefs could be interpreted as judgment errors, most were credible on a personal level. CONCLUSIONS: Most men who underwent PSA counseling cited underlying beliefs rather than the content of counseled information as the basis for their decisions regarding future PSA screening. If widespread, such beliefs may render clinician counseling and decision support methods less effective. Eliciting patient beliefs prior to counseling may improve the shared decision-making process. PMID- 12088292 TI - Tobacco-control policies in 11 leading managed care organizations: progress and challenges. AB - CONTEXT: Although evidence-based national guidelines for tobacco-dependence treatment have been available since 1996, translating these guidelines into clinical practice is challenging. PRACTICE PATTERN EXAMINED: Policies regarding tobacco-dependence treatment (e.g., written guidelines and coverage of pharmacotherapy) and implementation strategies of 11 U.S. managed care organizations known to have strong tobacco-control programs. DATA SOURCES: Detailed telephone interviews with multiple informants at each health plan and review of written treatment guidelines and policies for tobacco dependence. RESULTS: Although 10 of 11 plans had adopted tobacco-dependence treatment guidelines consistent with the national guideline, fewer had guidelines for special groups, such as adolescents (6 plans), parents (5 plans), pregnant women (5 plans), and hospitalized smokers (3 plans). Most plans offered clinician training and recommended office systems to support provider efforts; however, fewer actively facilitated their implementation. Most plans provided other support for tobacco treatment, including dedicated budgets, designated staff, and an oversight committee. All plans offered some coverage for tobacco-cessation pharmacotherapy and behavioral counseling, although not to the extent recommended by the national guideline. CONCLUSION: Implementation of national tobacco treatment guidelines is feasible in closed-panel managed care organizations. However, even these leading health plans could do more to comply with national practice guidelines on tobacco-dependence treatment and make it easier for clinicians to help patients stop smoking (e.g., through increased training and expanded coverage of tobacco-dependence treatment). PMID- 12088293 TI - Prostate biopsies in men with limited life expectancy. AB - CONTEXT: Authorities discourage prostate screening in men who are likely to die from causes other than prostate cancer. PRACTICE PATTERN EXAMINED: Use of prostate biopsy-a proxy for screening-in men aged 65 and older with limited life expectancy (i.e., estimated to be less than 10 years). DATA SOURCE: Five percent samples of Part A (hospital) and Part B (physician) Medicare claims for 1993 through 1997. RESULTS: 22% of all Medicare beneficiaries who underwent a prostate biopsy had a limited life expectancy, corresponding to a rate of 1420 biopsies per 100,000. This rate did not change significantly between 1993 and 1997. For men with a life expectancy greater than 10 years, the biopsy rate was 2,360 per 100,000. Among men with limited life expectancy, in the year following the biopsy, 1.6% had radical prostatectomy and 2.3% had external-beam radiation. Thirty-nine percent were hospitalized. CONCLUSION: A substantial proportion of prostate biopsies are being performed in men with a life expectancy of less than 10 years. These men are unlikely to benefit from the biopsy or subsequent treatment. PMID- 12088295 TI - Key challenge for antitobacco activists. PMID- 12088296 TI - Surgical management of pancreatic cancer. AB - Pancreatic cancer is the fifth leading cause of cancer death in the United States, with an overall survival rate of 3%. Unfortunately, only a minority of patients present with localized disease amenable to surgical resection. Over the past 20 years, improvements in operative and perioperative management have led to a decrease in operative mortality, shorter hospital stay, and overall 5-year survival of approximately 20% in patients undergoing pancreaticoduodenectomy. Despite advances in nonoperative palliation, surgery continues to play an important role in the management of patients with locally advanced, unresectable pancreatic cancer. PMID- 12088294 TI - The healthcare cost and utilization project: an overview. AB - DATABASE: . Healthcare Cost and Utilization Project (HCUP)-a family of databases including the State Inpatient Databases (SID), the Nationwide Inpatient Sample (NIS), the Kids' Inpatient Database (KID), and the outpatient databases State Ambulatory Surgery Data (SASD) and State Emergency Department Data (SEDD). DESCRIPTION: Multistate, inpatient (SID, NIS, KID) and outpatient (SASD, SEDD) discharge records on insured and uninsured patients. SOURCE: Partnership between the Agency for Healthcare Research and Quality (AHRQ) and public and private statewide data organizations. AVAILABLE DATA: Selected data elements from inpatient and outpatient discharge records, including patient demographic, clinical, disposition and diagnostic/procedural information; hospital identification (ID); facility charges; and other facility information. DATA YEARS AVAILABLE: Varies by database: NIS 1988-2000; SID 1995-2000; KID 1997 and 2000; SASD 1995-2000; and SEDD in pilot phase. Future data years anticipated for all datasets and back years for SID and SASD. UNITS OF ANALYSIS: Patient (in states with encrypted patient identification), physician, market, and state. RESEARCH QUESTIONS: Quality assessment, use and cost of hospital services, medical treatment variations, use of ambulatory surgery services, diffusion of medical technology, impact of health policy changes, access to care (inference), study of rare illness or procedures, small area variations, and care of special populations. STRENGTHS: Largest collection of all-payer, uniform, state-based inpatient and ambulatory surgery administrative data. LIMITATIONS: Lacks clinical detail (e.g., stage of disease, vital statistics) and laboratory and pharmacy data. Ability to track patients across time and setting varies by state. ACCESS TO DATA: Access available to all users who sign and abide by the Data Use Agreement. Application kits available at www.ahrq.gov/data/hcup. HCUPnet, an on line interactive query tool, allows access to data without purchase (www.ahrq. gov/data/hcup/hcupnet.htm). PMID- 12088298 TI - Clinical trials referral resource. Current clinical trials in non-Hodgkin's lymphoma. PMID- 12088297 TI - Treatment of dyspnea in cancer patients. AB - Dyspnea is defined as a sensation of difficult or uncomfortable breathing. The symptom is highly prevalent among cancer patients with and without direct lung involvement. The gold standard of assessment is based on patient self-report. Objective measures such as respiratory rate, oxygen saturation, and arterial blood gas measurements frequently do not correlate with the subjective experience of dyspnea. Consistent with patient goals and the disease context, treatment should be directed at removing the underlying cause when possible. Fast, safe, and effective symptomatic relief of dyspnea is possible whether or not identifiable reversible causes exist. In fact, relieving symptoms can be considered in conjunction with treating specific reversible causes. Opioids are the first-line therapy for relief of dyspnea symptoms. When prescribed appropriately, respiratory depression is not a significant concern. In the rare cases in which opioids are unable to control dyspnea, sedation is an effective, ethical, and legal option. PMID- 12088299 TI - Update on radiation therapy for endometrial cancer. AB - The best clinical outcomes for patients with endometrial cancer seem to be achieved with either surgery alone or a combination of surgery and radiation therapy. Although once administered preoperatively, irradiation is now rarely given prior to surgery in this population. After surgical staging, most patients receive postoperative adjuvant therapy based on their pathologic risk factors. Although prospective randomized trials in these patients are limited, recent studies have attempted to determine the best management strategies for the disease. Based on these investigations, treatment recommendations are outlined for patients who are surgically staged and for those with incomplete surgical staging. Also described is the use of irradiation in recurrent endometrial cancer after surgery alone, as well as palliative radiation. In addition, ongoing prospective randomized trials are described. PMID- 12088300 TI - The health economics of palliative care. AB - Only a few studies have assessed the economic outcomes of palliative therapy. The major areas of interest include hospice care, the process and structure of care, symptom management, and palliative chemotherapy compared to best supportive care. Compared with nonhospice care, hospice care saves at best 3% of total care costs. Advance directives done early in the disease course may save end-of-life care costs, but when done in the hospital do not save money or influence care choices. Nurse coordination of palliative care maintained clinical outcomes of dying patients and saved 40% of costs. A structured ethics review of those likely to die in the intensive care unit also appears to match the type of care to the outcome, and save costs. There are remarkably few randomized clinical trials of pain and symptom control interventions in end-of-life care, so few conclusions can be drawn about current treatments. There are no examples of chemotherapy that save money compared to best supportive care. Current data suggest that changes in palliative care cost can only come from dramatic changes in how we provide care. One model is coordinated, expert, high-volume care that can prevent end-of-life hospitalization, with early use of advance directives. Preliminary data from our program support the hypothesis that costs may be reduced by 40% to 70%. PMID- 12088301 TI - Smoking cessation counseling needed with spiral CT screening for lung cancer. PMID- 12088302 TI - Current perspectives on pain in AIDS. AB - As active participants in the care of patients with acquired immunodeficiency syndrome (AIDS), oncologists need to be aware of the many facets of pain management in this population. This two-part article, which will conclude in the July 2002 issue, describes the prevalence and types of pain syndromes encountered in patients with AIDS, and reviews the psychological and functional impact of pain as well as the barriers to adequate pain treatment in this group and others with human immunodeficiency virus (HIV)-related disease. Finally, principles of pain management, with particular emphasis on controlling pain in HIV-infected patients with a history of substance abuse, are outlined. PMID- 12088303 TI - Meetings, journals, and the internet: networks of communication. PMID- 12088304 TI - Threshold electrical stimulation (TES) in ambulant children with CP: a randomized double-blind placebo-controlled clinical trial. AB - A randomized double-blind placebo-controlled clinical trial was carried out to determine whether a group of stable children with cerebral palsy (36 males, 21 females; mean age 10 years 11 months, range 5 to 18 years) would improve their motor skills after 12 months of threshold electrical stimulation (TES). Two thirds received active and one third received inactive stimulators. For the primary outcome we constructed a set of plausible motor function tests and studied the change in summary indices of the performance measurements. Tests were videotaped and assessed blindly to record qualitative changes that might not be reflected in performance measurements. We also judged range of motion, degree of spasticity, and muscle growth measured by CT. Fifty seven of 82 outpatients who were able to walk at least with a walker, completed all 12 months of treatment (hemiplegia n=25, diplegia n=32). There was no significant difference between active and placebo treatment in any of the tested groups, nor combined. Visual and subjective assessments favoured TES (ns), whereas objective indices showed the opposite trend. We conclude that TES in these patients did not have any significant clinical effect during the test period. PMID- 12088305 TI - Visual-perceptual impairment in a random sample of children with cerebral palsy. AB - Several studies have tried to establish the prevalence of visual-perceptual impairment in children with physical disabilities, particularly in those with cerebral palsy (CP), but failed to take into account the selective impairment of non-verbal intelligence that is frequent in these children. This has resulted in the confounding of visual-perceptual and non-verbal intelligence impairment. In the present study we aimed to determine how widespread visual-perceptual impairment is in children with CP by evaluating perceptual ability together with the performance level on non-verbal intelligence subtests. All children (n=96; 44 females, 52 males) who attended an institute for children with physical disabilities were included (age ranged from 4 years 11 months to 21 years 5 months) who had a non-verbal mental age between 3 and 7 years; Total IQ was <85 in 91% of participants. They were given a grating acuity task and the visual perceptual battery L94, comprising six visual object recognition and two visuoconstruction tasks. Relative to their performance level on non-verbal intelligence subtests, 37.5% of the children were impaired on at least one task, and 18.7% on two or more tasks. No child was impaired on the visuoconstruction tasks. Visual-perceptual impairment was highest among six children with brain malformation (67%), followed by spastic CP (40%), and brain damage acquired after the first year of life (38%). There was no difference in visual-perceptual impairment between the subtypes of spastic CP. Results are not secondary to visual acuity deficits, as only one L94 task was significantly correlated with acuity impairment. We conclude that visual-perceptual impairment is frequent in children with physical disabilities, and not restricted to children with CP of hypoxic-ischaemic origin. PMID- 12088306 TI - Children with ADHD and motor dysfunction compared with children with ADHD only. AB - The purpose of this study was to identify group differences in children with attention-deficit-hyperactivity disorder and motor dysfunction (ADHD-MD) and ADHD only, and to evaluate the medication responsiveness of ADHD-MD. Sixty-three children (49 males and 14 females; mean age 9 years 10 months, SD 2 years 10 months) underwent a triple blind, placebo-controlled crossover study evaluating two dose levels of methylphenidate (0.3 mg/kg and 0.5 mg/kg [DOSAGE ERROR CORRECTED], twice daily) and placebo. Forty-nine trials were completed. Nineteen were children with ADHD-MD, 44 had ADHD only. Behavior and functioning were assessed at home and at school. Treatment effects were assessed using the Abbreviated Symptom Questionnaire for Parents and Teachers. Children with ADHD-MD were more likely to have severe ADHD-combined type and other neurodevelopmental and behavioral problems. Both groups of children had a linear dose response to medication (placebo, low, high) and there was no evidence of a group by dose interaction or an overall group effect at home or school. The lack of group effect suggests that these children responded to medication like the other subgroups. PMID- 12088307 TI - The BREV neuropsychological test: Part I. Results from 500 normally developing children. AB - The Battery for Rapid Evaluation of Cognitive Functions (Batterie Rapide d'Evaluation des Fonctions Cognitives: BREV) was designed to provide health professionals with a quick clinical tool for screening acquired and developmental cognitive deficits in children aged 4 to 8 years. The BREV explores oral language in both its expressive and receptive forms, non-verbal functions, attention, verbal and visuo-spatial memory, and main learning acquisition. Results of the first phase of validation are presented in this report consisting of internal validity measurements gained by testing 500 normally developing school children (257 females, 243 males; mean age 6 years 7 months, SD 1 year 6 months. The validation provides appropriate values for each of the 17 subtests assessing cognitive functions (oral language, non-verbal abilities, attention and memory, educational achievement) in 10 age groups, from 4 to 8 years of age. All subtests with the same content for any age revealed values which increased significantly with age. Interreliability was tested in a retest for 70 children and scores obtained on retesting correlated significantly with initial values. The BREV is a reliable test with carefully established normative values, appropriate for preschool and school-age children. PMID- 12088308 TI - The BREV neuropsychological test: Part II. Results of validation in children with epilepsy. AB - The Battery for Rapid Evaluation of Cognitive Functions (Batterie Rapide d'Evaluation des Fonctions Cognitives: BREV) is a quick test to screen children with higher-functioning disorders and to define the patterns of their disorders. After standardization tests in 500 normally developing children aged 4 to 8 years, validation consisted of comparative evaluation of the specificity and sensitivity of the BREV with a wide reference battery in 202 children with epilepsy (108 males, 94 females; mean age 6 years 6 months, SD 1 year 8 months). Children were divided into 10 age groups from 4 to 8 years of age and represented eight epileptic syndromes. The reference battery included verbal and non-verbal intelligence assessment using the Wechsler scale, oral language assessment with a French battery for oral language study, drawing with the Rey figure, verbal and visuo-spatial memory with the McCarthy scale subtest and the Rey figure recall, and educational achievement with the Kaufman subtests. Every function evaluated with the BREV was significantly correlated with the reference battery testing a similar function (p=0.01 to 0.001). Specificity and sensitivity of the BREV verbal and non-verbal scores were correlated with those of the Wechsler scale in more than 75% of children. The BREV, therefore, appears to be a reliable test which has been carefully standardized and validated and is valuable in screening for cognitive impairment in children. PMID- 12088309 TI - Infant sucking ability, non-organic failure to thrive, maternal characteristics, and feeding practices: a prospective cohort study. AB - This prospective study examined the relation of neonatal sucking to later feeding, postnatal growth, maternal postpartum depression, and feeding practices. Healthy infants of at least 37 weeks gestational age were recruited. At 1 week of age, a strain-gage device was attached to the infant's cheeks during sucking to identify sucking efficiency. Two-hundred and two infants (100 males, 102 females; mean age 39.6 weeks, SD 1.1 weeks) with efficient sucking and 207 (101 males, 106 females; mean gestational age 39.4 weeks, SD 1.2 weeks) with inefficient sucking were identified. Growth was measured at 2, 6, 10, and 14 months. Mothers completed a feeding questionnaire and the Edinburgh Postnatal Depression Scale at the same testing points. While 18 infants (5%) showed a downward shift in growth, their clinical picture did not present as non-organic failure to thrive (NFTT). Inefficient neonatal sucking did not predict postnatal growth, later feeding difficulties, nor maternal feeding practices, but concurrent inefficient feeding did. Maternal depression did not affect feeding practices, infant feeding abilities, nor growth, suggesting that the importance of maternal postpartum depression in association with feeding may be less than previously assumed. The term NFTT, therefore, merits reexamination and a more focused definition. PMID- 12088310 TI - Transient dystonias revisited: a comparative study of preterm and term children at 2 1/2 years of age. AB - Minor neurological dysfunctions (MND) have frequently been reported as an outcome of preterm birth. Behavioural and learning difficulties are a known feature, but coordination problems are especially described in preterm infants at later ages. All preterm infants in our study were born with a gestational age of <32 weeks and a birthweight of <1500 g. The aim of the study was to determine if in a normal clinical situation, children at risk for developing coordination problems could be detected by focusing particularly on their spontaneous, although elicited, motor performance. Forty-two children of 2 years 6 months of age were studied of whom 25 were 'low-risk' preterm and 17 were born at term. All children had been followed up since birth. In a structured, free-field situation the quality of body rotation, spontaneous reaching-out, and manipulation during parts of the Bayley Developmental Test were observed and videotaped. Body rotations were scored on a 2-point scale, and arm and hand functions on a 3-point scale. Most preterm infants showed non-optimal body rotations and borderline or non optimal arm and hand functions in contrast to most term children. A clear connection was found between less optimal body rotations and poor arm and hand functions. Retrospectively, poor arm and hand functions at 2 years 6 months seemed to be related to those at 39 weeks. At the age of 39 weeks a clear relation had been found between poor postural control (many preterm infants could not sit independently) and earlier hyperextension of the trunk at 18 weeks and quality of arm-hand functions. In the present study at 2 years 6 months a correlation with the former hyperextension could no longer be found, but there was a significant relation between poor trunk rotation and arm and hand function and the earlier arm and hand functions at 39 weeks. PMID- 12088311 TI - Two siblings with a new Aicardi-Goutieres-like syndrome. AB - We present two siblings (male and female) with very similar characteristics comprising dysmorphic features, severe developmental delay, progressive microcephaly, tonic seizures, and hypothyroidism. The male also had micropenis and cryptorchidism. Both children developed pericardial effusions which caused the death of the female at age 16 months. The male's cardiac function was stable at last follow-up at the age of 15 months. Cerebral imaging showed widespread intracranial calcifications, delay in myelination, hypoplasia of the corpus callosum, and cerebral atrophy. CSF examination showed normal CSF white-cell count and was negative for interferon, although a cytotoxic antibody was thought to be present. Other causes of a neurodegenerative condition and congenital infection were excluded. The combination of these features has not been described before. We believe that these patients represent a new syndrome which has some of the features of Aicardi-Goutieres syndrome but is distinct from it. PMID- 12088313 TI - Neurodevelopmental treatment for cerebral palsy: is it effective. PMID- 12088312 TI - Folic acid and prevention of birth defects. PMID- 12088314 TI - Organizational development of veterinary radiology in the United States and Europe. PMID- 12088316 TI - A new device for stereotactic CT-guided biopsy of the canine brain: design, construction, and needle placement accuracy. AB - An inexpensive device was created for computed tomographic (CT)-guided stereotactic biopsy of the canine brain. The accuracy of the device was tested using 16, formalin-perfused, canine head specimens. For each dog, a 6-inch biopsy needle was guided into pituitary gland and caudate nucleus targets. Needle tracks were measured using the CT computer and infused with tissue staining solution. Hit success and actual needle track lengths were determined from sliced brain specimens. The device enabled accurate orientation and placement of the canine head in the slice plane, such that progressive penetration of the biopsy needle could be monitored. The caudate nucleus was hit 12/16 times (75% accuracy) and the pituitary gland 15.5/16 times (98.6% accuracy). Hit proportions for the two targets did not differ (P < 0.05). A significant difference was found between CT and actual track length for both targets (P < 0.01). This was attributed to incomplete staining of the bevel portion of the needle track. PMID- 12088315 TI - Accuracy of survey radiographic diagnosis of intervertebral disc protrusion in dogs. AB - To assess the diagnostic accuracy of survey radiography for canine thoracolumbar intervertebral disc protrusion, survey radiographs (lateral and ventrodorsal) of 64 dogs with surgically-confirmed thoracolumbar intervertebral disc protrusion, 51 dogs with negative myelograms and 29 dogs with various spinal conditions other than disc protrusion were reviewed by three independent observers who were unaware of any clinical information. There were marked differences in observer performance for diagnosis of intervertebral disc protrusion, although there were no significant differences in intraobserver diagnostic accuracy for small vs. large dogs. Accuracy of observers for determining sites of intervertebral disc protrusion using survey radiography was in the range 51-61%. All observers had low accuracy for identification of second sites of intervertebral disc protrusion. The most useful radiographic sign, narrowed intervertebral space, had only moderate sensitivity (range 64-69%) and moderate predictive value (range 63 71%) for intervertebral disc protrusion. Vacuum phenomenon was an infrequent but accurate sign of intervertebral disc protrusion. Recognition of multiple radiographic signs of intervertebral disc protrusion at one site was associated with increased accuracy of diagnosis. No observer was accurate enough to justify attempting targeted surgical treatment of intervertebral disc protrusion without myelography. PMID- 12088317 TI - Radiographic diagnosis--right kidney herniation in a cat. PMID- 12088318 TI - Changes in the canine femoropatellar joint space in the postsurgical, cruciate deficient stifle joint. AB - In this study, we investigate the factors that influence changes in articular cartilage thickness in naturally occurring osteoarthritis of the canine stifle joint. Fifty-eight dogs with QA secondary to cranial cruciate ligament deficiency were examined radiographically at 0, 7, and 13 months using 90 degree flexed mediolateral radiographs of both stifle joints. The femoropatellar joint space widths were measured using a computer-assisted video image analysis system. The femoropatellar joint space widths were found to be significantly increased in index joints as compared to contralateral joints. In addition, there was a significant increase in femoropatellar joint space (FPJS) width over the course of the study, particularly between entry and 7 months. Factors that might influence the degree of femoropatellar joint space increase (bodyweight, age, sex, disease duration, radiographic score) were investigated using regression analysis. Body weight at baseline showed a moderate to strong relationship with change in FPJS between 0 and 7 months (Pearson's partial correlation r = 0.417, p = .006). This relationship was less strong for change in femoropatellar joint space between 7 and 13 months (r = 0.324, p = .036). PMID- 12088319 TI - Meniscal ossicles in large non-domestic cats. AB - Radiographs of the stifles of 6 species of 34 large, non-domestic cats were reviewed foremost for the presence of meniscal ossicles and then for the presence of the other potential four sesamoids. The animals in the review included 12 lions, 7 tigers, 7 cougars, 3 leopards, 3 bobcats, and 2 jaguars. Fluoroscopy, arthrography, computed tomography, necropsy, and histology were also used to evaluate the stifles of one tiger after euthanasia. Ossicles were found in the region of the cranial horn of the medial meniscus in most of the lions, tigers, leopards, and jaguars. These ossicles were found in half of the cougars but in none of the bobcats. Among the large, non-domestic cats, meniscal ossicles had been reported previously only in Bengal tigers. The lions, tigers, and leopards having meniscal ossicles appeared to have a lateral but often not a medial fabella of the gastrocnemius muscle, an observation previously unreported. Popliteal sesamoids and patellas were present in all the skeletally mature cats. PMID- 12088320 TI - An intrathoracic malignant peripheral nerve sheath tumor in a dog. AB - A 7-year-old, neutered male Labrador Retriever presented for a persistent, productive cough and regurgitation. Radiography, ultrasonography, and computed tomography confirmed a large, smoothly marginated intrathoracic mass causing tracheal compression. The mass was removed via a thoracotomy, and a malignant peripheral nerve sheath tumor, most likely originating from the ventral spinal nerve roots, was confirmed using immunohistochemistry. PMID- 12088321 TI - Temporomandibular joint morphology in Cavalier King Charles Spaniels. AB - Temporomandibular joint dysplasia has been reported in several breeds of dog. Three Cavalier King Charles Spaniel (CKCS) skulls were examined and radiographed during the course of a separate study and all demonstrated changes consistent with bilateral temporomandibular joint dysplasia. Subsequently, skull radiographs from all CKCS dogs examined at the University of Glasgow Veterinary School between 1991 and 2001 were reviewed (n = 26). Only two of these dogs were radiographed specifically for investigation of the temporomandibular joint, although varying degrees of dysplasia were identified in all dogs where the joints were adequately visualized (n = 20). The head of four CKCS cadavers was also radiographed, and similar changes were found. This finding suggests that temporomandibular joint dysplasia is a widespread asymptomatic condition in the CKCS and should be regarded as a normal morphologic variation rather than a pathologic anomaly. Subtle changes are best seen on lateral oblique radiographs, although marked changes are also visible on dorsoventral views. The rotational angle or angle of articulation of each of the dysplastic mandibular condyles was measured and was related to the severity of the dysplastic changes. However, there was overlap between the values calculated for these abnormal joints and normal ones in other breeds, suggesting this measurement was of limited significance and the shape of the components of the temporomandibular joint are more relevant when assessing this joint for the presence of dysplastic changes. PMID- 12088322 TI - Radiographic appearance of bronchoalveolar carcinoma in nine cats. AB - Thoracic radiographs of nine cats with confirmed bronchoalveolar carcinoma (BAC) were reviewed retrospectively. Radiographic appearance of BAC was divided into three categories: mixed bronchoalveolar pattern, ill-defined alveolar mass, or mass with cavitation. In addition to these radiographic signs, all nine cats had evidence of some form of bronchial disease. Cavitary lesions were the most common finding (n = 5). In addition, three cats in this category had diffuse bronchointerstitial opacity and one cat had focal peribronchial cuffing. Five cats had either a mixed bronchoalveolar pattern with bronchiectasis (n = 3) or an ill-defined alveolar mass with peribronchial cuffing (n = 2). One cat had both a mixed bronchoalveolar pattern and a cavitary mass. Each of these nine cats had some form of bronchial disease (bronchointerstitial pattern, peribronchial cuffing, or bronchiectasis), which aids in the radiographic diagnosis of bronchoalveolar carcinoma and may represent airway metastasis. PMID- 12088325 TI - Diagnosis of urinary bladder rupture using ultrasound contrast cystography: in vitro model and two case-history reports. AB - Because urinary bladder rupture can be life threatening, a simple, safe technique for evaluating patients is desirable. Current diagnostic protocols involve radiographic imaging, but ultrasound-based contrast techniques have not been methodically evaluated in veterinary patients with urologic trauma. Ultrasound contrast cystography (contrast cystosonography) involves infusion of microbubbled saline solution through a urinary catheter. It was performed in an in vitro model and in 2 dogs with naturally occurring urinary bladder rupture. A positive result consisted of visualizing microbubbles sonographically in fluid surrounding the bladder immediately after infusion of contrast into the urinary catheter. A positive result was obtained both in the in vitro model and in the 2 dogs, with radiographic and surgical confirmation of naturally occurring intraperitoneal urinary bladder rupture in the dogs. Based on the results of this study, ultrasound contrast cystography appears to be more sensitive than two-dimensional (2D) abdominal sonography for detecting naturally occurring urinary bladder rupture in dogs. PMID- 12088323 TI - Radiographic diagnosis: intramedullary extrusion of an intervertebral disc. PMID- 12088326 TI - The scintigraphic anatomy of the equine sacroiliac joint. AB - The aim of this study was to describe the scintigraphic appearance and location of the sacroiliac joint (SI-joint) in dorsal and dorso 30 degree lateral views of the equine pelvis in a scintigram. The pelvis from 10 different horses was prepared by attaching plastic tubes along relevant margins of the bone and around the SI-joint. A radioactive solution was injected into the tubes, and scintigraphic images were acquired with a gamma camera. Five specific landmarks were identified on the left and right sides of the pelvis. There was no significant difference in the distance measurements between the left and right sides in the dorsal view. The SI-joint was roughly ovoid in shape, with its long axis angled to the midline. The most craniolateral point of the SI-joint (Y) was located at the middle (SD = 0.04) of a line drawn from the caudal margin of the tuber sacrale (TS) to the craniolateral margin of the tuber coxa (TC), which was called TS-TC. In 85% of the specimens, the long axis of the SI-joint (SILA) was located caudal to or intersecting TS-TC, and the mean angle between SILA and TS TC was 16.4 degrees (SD = 5.6 degrees). The distance from the caudomedial point of the SI-joint (Z) to Y was 0.38 times (SD = 0.04) the length of TS-TC. The distance from the tuber sacrale to the medial border of the SI-joint (TS-X) was 0.15 times (SD = 0.02) the length of TS-TC. In the oblique view, the SI-joint appeared wider and was positioned more laterally compared to the dorsal view. The distance TS-Y increased to 0.61 times (SD = 0.046) TS-TC, but SILA/TS-TC was essentially unchanged. The mean angle between TS-TC and SILA increased to 20.9 degrees (SD = 5.6 degrees). TS-X increased to 0.31 times (SD = 0.037) TS-TC. The location of the SI-joint was very similar in all horses, which makes it possible to estimate the location of the SI-joint in a scintigram in which the tuber sacrale and the craniolateral margin of the tuber coxa are identified. PMID- 12088327 TI - Images in medicine. William's pear bile. PMID- 12088324 TI - Cellular features of sonographic target lesions of the liver and spleen in 21 dogs and a cat. AB - Target lesions are seen in ultrasound images of the liver or spleen as nodules or masses with a hypoechoic rim and a hyperechoic or isoechoic center. To assess the diagnostic significance of finding a target lesion, the cytologic and/or histopathologic findings were reviewed in a series of 21 dogs and a cat that had hepatic and/or splenic target lesions noted during abdominal ultrasonography. Twelve of 16 hepatic target lesions and 5 of 7 splenic target lesions were malignant. In this series, the finding of one or more target lesions in the liver or spleen had a positive predictive value for malignancy of 74%; for the finding of multiple target lesions in one organ, the positive predictive value for malignancy was 81%. Benign lesions associated with target lesions were nodular hyperplasia of the liver and spleen, pyogranulomatous hepatitis, cirrhosis, and chronic active hepatitis. PMID- 12088328 TI - Images from the 2001 ACVR certifying examination thorax section. PMID- 12088329 TI - Images from the 2001 ACVR certifying examination: artifacts section. PMID- 12088330 TI - Suggestibility and negative priming: two replication studies. AB - Research suggests that inhibiting the effect of irrelevant stimuli on subsequent thought and action (cognitive inhibition) may be an important component of suggestibility. Two small correlation studies were conducted to address the relationship between different aspects of suggestibility and individual differences in cognitive inhibition, operationalized as the degree of negative priming generated by to-be-ignored stimuli in a semantic categorization task. The first study found significant positive correlations between negative priming, hypnotic suggestibility, and creative imagination; a significant negative correlation was obtained between negative priming and interrogative suggestibility, demonstrating the discriminant validity of the study results. The second study replicated the correlation between negative priming and hypnotic suggestibility, using a different suggestibility measurement procedure that assessed subjective experience and hypnotic involuntariness as well as objective responses to suggestions. These studies support the notion that the ability to engage in cognitive inhibition may be an important component of hypnotic responsivity and maybe of other forms of suggestibility. PMID- 12088331 TI - Hypnotic responsivity from a developmental perspective: insights from young children. AB - Evidence indicates that hypnotic responsivity in children younger than 8 years of age differs significantly from that of older children and adults. The sudden increase in responsiveness around age 8, differing patterns of item difficulty for young children, specific problems with hypnotic dream and age regression items, and the lack of conceptual distinction between volition and nonvolition argue for a fundamental discontinuity between young children and adults regarding responsivity. These differences result from underlying developmental processes that characterize young childhood, including limitations in executive cognitive functioning, more overt forms of involvement, and reliance on authoritative others for direction, regulation, and support. The unique features of young children's hypnotic responsivity offer the opportunity to reconsider hypnosis within a developmental context. PMID- 12088332 TI - Autonomic reactivity to cognitive and emotional stress of low, medium, and high hypnotizable healthy subjects: testing predictions from the high risk model of threat perception. AB - This study tested hypotheses derived from Wickramasekera's High Risk Model of Threat Perception (HRMTP) by comparing autonomic and affective responses to a cognitive and an emotional stress task in high, medium, and low hypnotizables. Electrodermal activity (EDA) was used as a measure of sympathetic activity, and the high frequency (HF) spectral component of heart rate variability as a measure of parasympathetic activity. High hypnotizables exhibited greater EDA at baseline and slower EDA recovery following both tasks than did medium and lows. Medium hypnotizables responded with greater decreases in normalized HF power than did highs and lows during the emotional stress task. The results suggest diminished EDA variability in high hypnotizables and the potential for HF power as an indicator of autonomic dysregulation in low and high hypnotizables, compared to mediums. The results are discussed in relation to predictions based on the HRMTP. PMID- 12088333 TI - The "big five" and hypnotic suggestibility. AB - A recent approach to personality measurement argues that the essential personality traits are encompassed by 5 basic factors: openness/intellect, conscientiousness, neuroticism, agreeableness, and extraversion. This study used the Big Five Inventory to test the hypothesis that 1 or more of the 5 factors underlie hypnotic suggestibility. No meaningful relationships between hypnotic suggestibility and any of the 5 factors were found. PMID- 12088334 TI - Hypnosis, the brain, and sports: Salient Findings July 2002. AB - Three particularly noteworthy articles addressing hypnosis have been published during the early portion of 2002. All, to a degree, address biological aspects of hypnotic response. One of these articles is a thoughtful summary and synthesis of neuroscience/hypnosis research to date, describing how neuroimaging techniques offer new opportunities to use hypnosis as a manipulation and to provide a means of studying hypnosis itself. A second article focuses on the physiology of sports and the usefulness of hypnosis in the practice of sport and exercise psychology. Finally, the third article describes a study of brain activation during actual and imagined handgrip during hypnosis. PMID- 12088335 TI - Expression of the vascular endothelial growth factor (VEGF) family in human endometrial blood vessels. AB - Endometrial regrowth is associated with intense angiogenesis, for which vascular endothelial growth factor-A (VEGF-A) is an important regulator. However, the expression of other members of the VEGF family is less well documented. The aim of this study was to localize members of the VEGF family (VEGF-A, -B and -C), and their receptors (VEGFR1, 2 and 3) in human endometrial blood vessels. Endometrial biopsies collected from four healthy and fertile women were used for immunohistochemistry assessments. Co-localization of VEGF-family proteins with CD34 stained endothelial structures was determined by image analysis. We demonstrate here the marked expression of VEGF-A as well as VEGFR2 and 3 in capillaries. Arterioles expressed VEGF-B, VEGFR1, 2, and 3 moderately and VEGF-A variably. Venules expressed only VEGFR3 markedly. In contrast, VEGF-C was not expressed in the arterioles, but moderately in the capillaries and weakly in the venules. VEGF-B was expressed in all blood vessels; however, VEGF-B was weakly expressed in capillaries and arterioles and moderately expressed in venules and arterioles. Thus, expression of VEGF-A. B and C and VEGF receptors 1-3 in endometrial blood vessels indicates a highly structured involvement of VEGF in the regulation of angiogenesis in the human endometrium. PMID- 12088336 TI - Serum tumor marker CA 125 for monitoring ovarian cancer during follow-up. AB - CA 125 is currently widely applied in the management of patients with ovarian cancer. However, a change in results of CA 125, which should be considered significant, has not been defined. The aim of this study was to investigate the ability of CA 125 to signal progressive ovarian cancer during follow-up after first-line chemotherapy. The study patients were selected retrospectively among 255 patients with stage IC-IV ovarian cancer. The evaluation of the CA 125 information was based on the analytical imprecision, the normal intra-individual biological variation, the sampling interval, and the cut-off value. Additionally, the utility of a new assessment criterion based upon an increment of 2.5 times the baseline CA 125 concentration confirmed by a third measurement was investigated. The efficiency of CA 125 to identify progression and non progression during follow-up varied between 76.5 and 79.9%, depending on the applied time limit for an acceptable positive lead time. The median lead time for true positive results was 95-99.5 days. Using the new elaborated criterion, the efficiency of CA 125 for identifying progression and non-progression varied between 75.7 and 78.5%, depending on the applied time limit for an acceptable positive lead time. The median lead time for true positive results was 91-95.5 days. CA 125 provided early and reliable information about progressive disease during follow-up. The applied criteria can therefore be recommended in further studies assessing the clinical utility of serological tumor markers in patients with ovarian cancer. PMID- 12088337 TI - Reference intervals for procalcitonin and C-reactive protein after major abdominal surgery. AB - Procalcitonin (PCT). a new marker proposed as a diagnostic tool for bacterial infections, triggers a systemic-inflammatory reaction in the body (sepsis, septic shock) and has potential use in a wide range of patient settings. To interpret the results from PCT measurements, we depend on reference intervals established from relevant populations. PCT and C-reactive protein (CRP) concentrations were analysed in 47 patients with a normal postoperative course after major abdominal surgery. The mean concentration of PCT declines from the first day and reaches half its initial values on the second day after the operation. whereas the mean concentration of CRP increases in the first 48 h and reaches half its maximum value on the fifth day after the operation. We present a continuous reference interval for plasma PCT and CRP concentrations in the first week following major abdominal surgery. For PCT we also present a graphic display of expected mean and expected upper reference limits predicted from the value measured on the first postoperative day. PMID- 12088338 TI - Blood coagulation and fibrinolysis in acute ischaemic and haemorrhagic (intracerebral and subarachnoid haemorrhage) stroke: does decreased plasmin inhibitor indicate increased fibrinolysis in subarachnoid haemorrhage compared to other types of stroke? AB - Ischaemic and haemorrhagic stroke may cause haemostatic abnormalities, apart from concomitant brain damage. In this study, some blood coagulation and fibrinolysis parameters were investigated in 30 patients with ischaemic stroke (atherothrombotic) and 30 with haemorrhagic (20 with intracerebral and 10 with subarachnoid haemorrhage) stroke. The following parameters were determined within the first 24h after stroke: prothrombin time (PT%). activated partial thromboplastin time (aPTT). fibrinogen, activity of FVII, antithrombin. plasmin inhibitor (PI) and fibrin D-dimer. Significant decreases in PT%, FVII activity and antithrombin as well as an increase in fibrinogen and D-dimer were noticed in ischaemic stroke and in both groups of patients with haemorrhagic stroke. PI levels were significantly lower in subarachnoid haemorrhage patients compared with those in controls and those in both the intracerebral haemorrhage and the ischaemic stroke patients. With the exception of this difference, there were no other differences between ischaemic stroke and the two types of haemorrhagic stroke. This could indicate that haemostatic abnormalities are a consequence of brain damage rather than primary haemostatic activation during thrombosis and/or bleeding in the acute phase of stroke. A decrease in the plasmin inhibitor could suggest excessive fibrinolysis in subarachnoid haemorrhage. PMID- 12088339 TI - Estimation of in vivo capillary or venous blood glucose concentration from analysis on stored venous blood or its plasma and use in quality control of near patient glucose tests. AB - Glucose concentrations were determined in capillary and venous blood and in venous plasma from 100 consecutive patients referred to an oral glucose tolerance test. The capillary blood was immediately transferred to a haemolysing and glucose stabilizing solution and frozen within 4h. Venous blood was drawn in heparin-sodium fluoride tubes and stored for 0, 4 or 24h at 4 or 20 degrees C. Aliquots of the venous blood were then treated in the same way as the capillary blood. The primary tubes were centrifuged, and aliquots of the plasma were stabilized, as described. All specimens from the same sampling event were analysed in the same analytical series on EBIO compact. Deming linear regression equations y = a + bx were calculated to estimate the glucose concentration in one specimen from that in a differently treated specimen e.g.: B(cB:fPt)---Glucose (0h) = 0.61 + 0.897*P(vB;fPt)-Glucose (24h. 20 C). n = 100, SD(y/x) = 0.25 mmol/L and B(vB) Glucose (0h) = 0.53 + 0.897*P(vB)--Glucose (24h. 20 degrees C), n = 196, SD(y/x) = 0.26 mmol,/L. The non-analytical part of the standard deviation of the ratio between the near-patient test result and its predicted value was 0.038 for both of the above predictions. In conclusion, the analytical (including sampling) variation of near-patient tests of glucose can be assessed by laboratory analysis on mailed, heparin-fluoride stabilized venous blood or on plasma samples made from the mailed blood. PMID- 12088340 TI - Leg gas exchange, release of glycerol, and uptake of fats after two minutes bicycling to exhaustion. AB - It is not known to what extent the muscles use fats and carbohydrates as substrate for oxidation after intense, anaerobic types of bicycling. Six healthy young men therefore bicycled at constant power for 2 min to exhaustion. Blood was drawn from indwelling catheters in the femoral artery and vein at intervals during the 1-h postexercise recovery. The blood samples were analysed for concentrations of O2 and CO2, and for free fatty acids (FFA), triacylglycerols (TG), and glycerol in plasma. The blood flow was also measured, and the rate of leg uptake of FFA, TG, and O2 and the release of CO2 and glycerol as well as its gas exchange ratio were calculated and integrated over the recovery period. The leg gas exchange ratio integrated over the exercise plus 1-h recovery period was 0.67 +/- 0.06 (mean +/- SEM ), suggesting pure fat oxidation. There was no statistically significant arterial-femoral-venous difference of FFA across the leg. The concentration of TG in plasma fell by 0.18 +/- 0.09 mmol L(-1) (32%) during the first 10 min of the recovery period, and the leg took up 18 +/- 8 micromol TG kg(-1) body mass (bm) during the whole 1-h recovery period. Free glycerol was released from the leg throughout the recovery period in excess of that released from hydrolysis of TG from plasma, suggesting that 30 +/- 10 micromol TG kg (-1) bm was hydrolysed, probably from intra-muscular stores. If fully oxidized, the triacylglycerols hydrolysed can account for 101% of the measured O2 uptake. Thus, muscle seems to use only triacylglycerols as substrate for its oxidative energy release after intense exercise. PMID- 12088341 TI - Levels of tissue inhibitor of metalloproteinases-1 in blood transfusion components. AB - Blood transfusion during surgery for solid tumors may reduce patient survival because of various bioactive substances present in blood preparations. The anti proteolytic protein tissue inhibitor of metalloproteinases-1 (TIMP-1) present in large quantities in platelets has been shown to stimulate cell growth and to inhibit apoptosis and may therefore be considered to influence tumor progression. We measured TIMP-1 levels in blood transfusion preparations. especially in platelet-containing preparations, before and after leucofiltration and at different time-points during storage. The mean TIMP-1 levels in whole blood (WB) and platelet-rich plasma (PRP) were slightly reduced by leucofiltration; WB: 41.6 microg/L versus 34.9 microg/L. PRP: 139.8 microg/L versus 127.2 microg/L. However, with prestorage leucofiltration. TIMP-1 levels in buffy-coat-derived platelet (BCP) pools were significantly reduced from 134.2 microg/L to 102.2 microg/L (p=0.0013). In saline-adenine-glucose-mannitol (SAG-M) blood preparations in which the platelet content is reduced by more than 99%,. TIMP-1 could not be detected. Extracellular TIMP-1 accumulated significantly in non filtered WB and in aferesis platelet concentrates (APC), but TIMP-1 was at no time detectable in SAG-M blood during storage. In conclusion. TIMP-1 is present in various platelet-containing blood preparations, but not in platelet-free preparations such as SAG-M, indicating that most of the TIMP-1 measured in blood preparations originates from platelets. Furthermore, TIMP-1 levels increased during storage in preparations containing platelets. which suggests a continuous disintegration of platelets. These data imply that information on preoperative blood transfusions should be taken into account when evaluating plasma TIMP-1 levels in patients. PMID- 12088342 TI - Assay of total antioxidant capacity: comparison of four methods as applied to human blood plasma. AB - Four spectrophotometric methods of determination of antioxidant capacity: a method based on the scavenging of the l,1-diphenyl-2-picrylhydrazyl (DPPH) free radical, the "ferric-reducing ability of plasma" (FRAP), a method based on reduction of the 2.2'-azinobis (3-ethylbenzthiazolinesulfonate) free radical (ABTS.+) and a kinetic method based on the oxidation of dihydro-2,7 dichlorofluorescein by 2,2'-azobis(2-amidopropane) (ABAP) were compared with respect to standard antioxidants (ascorbate, glutathione, Trolox and urate) and human blood plasma. Various reactivities of standard antioxidants in different tests were found. glutathione showing a low reactivity in the FRAP assay. Kinetic measurements show that the reduction of indicators, especially by blood plasma, may not be complete at recommended times of the assays and the time of measurement is an important parameter when comparing the results. PMID- 12088344 TI - The Human Genome Project and genome variation. PMID- 12088343 TI - Diagnostic efficacy of serum cross-linked N-telopeptide (NTx) and aminoterminal procollagen extension propeptide (PINP) measurements for identifying elderly women with decreased bone mineral density. AB - The purpose of this study was to determine the diagnostic sensitivity, specificity, predictive value and overall efficiency of serum cross-linked N telopeptides of bone collagen (NTx) and aminoterminal procollagen extension propeptide (PINP) measurements for identifying women with decreased spine, femoral neck and total body bone mineral density (BMD). Serum NTx and PINP levels and dual X-ray absorptiometry were performed on 196 healthy elderly women, aged 60-90 years. Twelve women were classified as having decreased BMD on the basis of regional and total skeletal densitometric values that were 1.5 to 2.5 standard deviations (SD) below the respective, age-stratified means and were compared with 184 women with BMD values greater than 1.5 SD below the mean. The results of receiver operating characteristic analysis revealed that a cutoff level of more than 15.0 nmol BCE/L for serum NTx, as measured by the Osteomark assay (Ostex International, Seattle WA USA) was associated with a 100% sensitivity and 70% specificity rate for identifying postmenopausal women with low BMD. The positive likelihood ratio was 3.3 and the negative predictive value was 1.0 using the 15.0 nmol decision level for NTx. The overall diagnostic efficiency of a single NTx measurement for identifying women with low BMD was 89%. A cutoff level of >45.0 microg/L for PINP as measured by the Orion Diagnostica RIA assay (Espoo, Finland) had a diagnostic sensitivity of 83% and specificity of 64% for identifying women with decreased BMD. The positive likelihood ratio was 2.3. the negative predictive value 0.98 and the overall diagnostic efficiency 73% using the 45.0 microg/L decision level for PINP. These results warrant future studies using larger populations that are inclusive of more women with low bone mineral density. PMID- 12088345 TI - Communicating in a multicultural society. II: Greek community attitudes towards cancer in Australia. AB - BACKGROUND: Open and full disclosure of information regarding diagnosis and prognosis is the prevailing approach to cancer patients. However, such a view appears contrary to the preferences of many ethnic groups in Australia. AIMS: This study sought to examine the range of attitudes to cancer, its treatment and disclosure of information among unaffected Greek adults, as part of an ongoing project to develop culturally appropriate cancer care in Australia. METHODS: Respondents were recruited from first generation Australian residents. Twenty nine men and 29 women, half over and half under the age of 60 years, participated. Eight focus groups were conducted by a bilingual facilitator and supplemented with eight individual face to face interviews. RESULTS: Several areas of misunderstanding were identified concerning the causes and outcomes of cancer. Having a cancer was regarded as a source of shame. Disclosure of diagnosis, but less so prognosis, was favoured, and only to immediate family members. Family members translating for the doctor were reported to commonly alter or 'soften' the doctor's message without the patient's knowledge in order to protect the patient. Greek doctors were favoured, and open discussion of alternative medicines was sought. CONCLUSIONS: The Greek community is the most established migrant community in Australia, but clearly many of their attitudes to cancer are at variance with what is considered good practice by clinicians. An increased awareness of cultural differences is needed to achieve optimal health outcomes in the diverse communities that make up modern Australia. PMID- 12088346 TI - Sleep disorders: a potential role in New Zealand motor vehicle accidents. AB - BACKGROUND: The potential role of sleep disorders in New Zealand motor vehicle accidents (MVA) has not been systematically studied. AIMS: To trial a recruitment method and document the frequency of sleep disorders in drivers attending an emergency department with injury following MVA. METHODS: Injured drivers admitted to Wellington Hospital Emergency Department were prospectively recruited by letter and follow-up telephone call. Data from a traffic accident, health and sleep questionnaire and polysomnography were collected and analysed using the SPSS statistical package. RESULTS: Of 120 eligible drivers, 40 (33%) completed the study protocol. The mean age of participants was 44 +/- 17 years. Non participants were younger (mean age 33 +/- 14 years, P < 0.01) and more often of Asian or Pacific Island ethnicity (P < 0.01). Twenty-seven (67%) felt unrefreshed on waking and 10 (25%) had an elevated Epworth Sleepiness Score (> 10/24). Six (15%) felt drowsy or fell asleep prior to the accident. Sleep disorders were common (40%): obstructive sleep apnoea 14 (35.9%), periodic limb movements of sleep three (7.7%), sleep restriction (< 6 h sleep on a regular basis) five (12.5%), and insomnia one (2.5%). CONCLUSIONS: Daytime somnolence and sleep disorders were commonly found in drivers attending the Emergency Department after accidents resulting in injury. Driver fatigue and sleep disorders should be considered as a potential contributing factor in New Zealand MVAs. Recruitment of drivers following a MVA via the Emergency Department is challenging. Strategies to improve recruitment are discussed. PMID- 12088347 TI - Medications taken by 26-year-olds. AB - BACKGROUND: Most descriptive pharmacoepidemiological reports are from studies of older people, and little is known about medication use by younger adults. AIMS: The aim of the present study was to examine the prevalence of medication usage in the previous 2 weeks among young adults. METHODS: Detailed information was collected on medications taken by 26-year-old participants in the longstanding Dunedin Multidisciplinary Health and Development Study. RESULTS: At age 26, 980 (96.2%) of the surviving cohort participated; medication data were available for 978 individuals, of whom 78.1% had taken one or more prescribed or over-the counter medications. Medication use was higher among females. The most prevalent drug categories were analgesics (taken by 22.8% of the sample), hormonal contraceptives (21.9%; 44.7% of females), nutrient supplements (17.6%) and anti asthma drugs (11.1%). CONCLUSIONS: The prevalence and pattern of medication use among younger adults differ from those reported for older age groups. Asthma appears to be the most prevalent chronic medical condition requiring pharmacological intervention (and for which medical care has been sought) among younger adults. The low rates of use of antidepressants and anxiolytics suggest that common psychiatric disorders may currently go unrecognized or untreated among young adults, and the high prevalence of 'third generation' oral contraceptive use among females is of particular concern. PMID- 12088348 TI - Lack of clinical utility of the Siriraj Stroke Score. AB - BACKGROUND: The ability to distinguish between infarct and haemorrhage is essential to the management of acute cerebrovascular disease. In hospitals where emergency neuroimaging is not available, the use of stroke scores has been proposed to distinguish ischaemic from haemorrhagic stroke. AIMS: To determine the accuracy of the Siriraj Stroke Score in distinguishing ischaemic from haemorrhagic stroke in a cohort of Chinese patients. METHODS: We prospectively assessed and calculated the Siriraj Stroke Score from 253 patients with acute stroke. The sensitivity, specificity, positive and negative predictive values of this score were determined. RESULTS: For cerebral haemorrhage, the sensitivity and specificity were both 90% or above, but the positive predictive value was not greater than 70%. For cerebral infarct, the sensitivity and specificity were around 80%, while the positive predictive value exceeded 90%. Analysis by plotting receiver operating characteristic curves failed to find other cut-off points that would improve the performance of the Siriraj Stroke Score. CONCLUSIONS: Considering the inconsistent results from this study and previous studies of the Siriraj Stroke Score, we suggest that scoring systems that only require a small number of variables are unlikely to achieve the level of accuracy needed for clinical decision-making. PMID- 12088350 TI - Feminism and medicine. PMID- 12088349 TI - Audit of the management of suspected giant cell arteritis in a large teaching hospital. AB - BACKGROUND: The diagnosis of giant cell arteritis (GCA) is often confirmed by an early temporal artery (TA) biopsy of adequate length. Treatment of this condition with high-dose corticosteroids may be associated with significant morbidity, including osteoporosis. AIM: To audit current management of patients with suspected GCA at Auckland Healthcare, a large teaching hospital. METHODS: We performed a retrospective chart review of all TA biopsies from January 1996 to June 2000. A total of 117 biopsies from 111 patients was audited. Of these patients, 37/111 (33%) had a final clinical diagnosis of GCA (GCA patients). The areas of interest for audit were waiting time for TA biopsy, length of sample, initial corticosteroid therapy and osteoporosis prophylaxis. RESULTS: The mean waiting time for biopsy for all patients was 5.6 days (range 0-42 days). This time varied from 9.3 days for rheumatology patients to 2.6 days for ophthalmology patients (P = 0.003). Only 44/117 (37.6%) specimens measured more than 10 mm. For GCA patients, the median initial oral prednisone dose was 60 mg/day. Osteoporosis prophylaxis was prescribed in 24/37 (65%) GCA patients, most commonly cyclical etidronate. CONCLUSIONS: There is significant variation in the management of GCA within our institution. This audit has highlighted several areas where improvement could be made, particularly in streamlining the process of obtaining TA biopsy and in promoting the use of osteoporosis prophylaxis. PMID- 12088351 TI - What are the barriers to clinical practice improvement? PMID- 12088352 TI - Hereditary colorectal cancer: keeping it in the family--the bowel cancer story. AB - Up to 20% of colorectal cancers are thought to have a genetic component. Several familial syndromes are known to confer an increased risk for colorectal cancer. Advances in our understanding of these syndromes has improved the care delivered to, and the overall survival of, these patients. Genetic testing has great potential to further improve detection and direct subsequent preventative measures. The diagnosis, management and surveillance issues relating to some of the more commonly encountered syndromes - in particular Familial Adenomatous Polyposis and Hereditary Non-Polyposis Colorectal Cancer - are reviewed. PMID- 12088353 TI - Why is quality assurance so difficult? A review of issues in quality assurance over the last decade. AB - The quality assurance movement has been unable to produce major improvements in the realm of clinical practice because of an inability to make satisfactory measurements of process and/or outcome, together with the intrinsic difficulties associated with producing change. Progress in both these areas is likely to be slow. Improvements in the quality of care occurring as a result of the introduction of information technology into clinical practice may be seen more quickly. PMID- 12088354 TI - Optimizing glycoprotein IIb/IIIa inhibition: lessons from recent randomized controlled trials. AB - Recent randomized clinical trials with intravenous glycoprotein IIb/IIIa inhibition have provided unanticipated results, thereby questioning their role as empirical medical management for acute coronary syndromes. The lack of benefit with abciximab observed in Global Utilization of Strategies to Open Occluded Coronary Arteries IV is somewhat inconsistent with the benefits seen with this agent in coronary intervention, and the benefits of an early invasive approach incorporating tirofiban seen within Treat Angina with Aggrastat and determine Cost of Therapy with an Invasive or Conservative Strategy Thrombolysis in Myocardial Infraction 18. Additionally, a direct 'head-to-head' comparative study of abciximab and tirofiban within the setting of percutaneous coronary intervention demonstrates clinically relevant superiority with abciximab with respect to 30-day outcomes. in the setting of coronary instability and mechanical plaque disruption is more complex than initially perceived. Hence, although an abundance of evidence details the efficacy of these agents, their optimal clinical application remains somewhat challenging in light of these recent data. However, within the context of previous trial experience, the current evidence highlights several key aspects of glycoprotein IIb/IIIa inhibitor therapy that may be associated with improved patient outcomes. In particular, these trials indicate: (i) the importance of selecting high-risk patients in whom substantial clinical benefit is evident, (ii) the incorporation of these agents into an early invasive strategy, thereby matching the timing of vascular injury with maximal platelet inhibition and (iii) optimal dosing to achieve the high levels of platelet inhibition that appear to be required for efficacy with these agents. PMID- 12088355 TI - Temozolomide-induced flare in high-grade gliomas: a new clinical entity. AB - A transient deterioration in neurological status following commencement of chemotherapy for high-grade gliomas has not been previously described. We report eight cases of transient deterioration following administration of temozolomide, a relatively new cytotoxic agent used in the treatment of high-grade gliomas. We believe this represents the novel clinical entity of temozolomide-induced tumour flare. PMID- 12088356 TI - Assessment and relevance of enzyme-linked immunosorbent assay for antibodies to Saccharomyces cerevisiae in Australian patients with inflammatory bowel disease. AB - The frequency of antibodies to Saccharomyces cerevisiae (ASCA) in Crohn's disease, ulcerative colitis and non-inflammatory bowel disease controls has been compared using two commercially available assays. The Medizym test resulted in sensitivity of 50% and specificity of 93% for Crohn's disease. The corresponding figures for the QUANTA Lite assay were 79% and 74%, respectively. Using ASCA and perinuclear antineutrophil cytoplasmic antibody (pANCA) in combination, the sensitivity and specificity of ASCA+/pANCA- for Crohn's disease using the Medizym kit were 50% and 100%, respectively, compared with 79% and 93% using QUANTA Lite. ASCA-/pANCA+ was 100% specific for ulcerative colitis with either assay. ASCA can be found in Australian patients with Crohn's disease at a similar frequency to that reported overseas. PMID- 12088357 TI - My left arm: experiencing brachial plexopathy. PMID- 12088358 TI - Severe hypokalaemia secondary to dicloxacillin. PMID- 12088359 TI - Ticlopidine-associated cholestatic hepatitis. PMID- 12088361 TI - Management of chronic obstructive pulmonary disease in the twenty-first century. PMID- 12088360 TI - Management of chronic obstructive pulmonary disease in the twenty-first century. PMID- 12088362 TI - Improvement of primary settling performance with activated sludge. AB - In biological treatment plants employing activated sludge processes, it is possible to recirculate some portion of the waste activated sludge that is not sent to the aeration basin, to the inlet of the primary sedimentation tanks. But in the literature there is no detailed information about the conditions, ratios and the characteristics of the waste sludge that can be recirculated back. However, depending on its settling characteristics, the addition of waste activated sludge to raw wastewater may improve primary settling. Settling tests have shown that the effect of waste activated sludge on primary settling is strongly dependent on the mean cell residence time (or sludge age), theta(c), of the waste activated sludge and also on the suspended solids concentration. Different sludge ages of 4, 6, 8, 10, 14, 20 and 26 days, and for each sludge age at least five different initial suspended solids concentrations were studied. A sludge age of 8-10 days achieved the optimum efficiency in terms of the remaining suspended solids concentration as well as percent-suspended solids removal. Also, the settled sludge volumes were measured throughout the experiments; so, the comparison was made between settled sludge volumes, initial suspended solids (SS) concentrations and theta(c). PMID- 12088363 TI - Ozonation of 1-naphthalene, 1,5-naphthalene, and 3-nitrobenzene sulphonic acids in aqueous solutions. AB - Ozone oxidation kinetics of 1-naphthalene (INS), 1,5-naphthalene (1,5NDS), and 3 nitrobenzene (3NBS) sulphonic acid is reported here. The contribution of indirect free-radical pathways and direct ozone reactions are accounted for in the proposed model. Second order kinetic constants of direct reactions were estimated at around 252 (M(-1)s(-1)), 41 (M(-1)s(-1)) and 22 (M(-1)s(-1)), for INS, 1,5NDS, and 3NBS sulphonic adds, respectively. At pH 3, the indirect reaction accounted for 2%, 15% and 4% of total primary oxidation of 1NS, 1,5NBS, and 3NBS sulphonic acids, respectively. At pH 9, indirect reaction contribution increased to 73%, 84% and 48%, respectively. C4 compounds (maleic and fumaric acids), C2 (oxalic), C1 (formic) and sulphate were identified as oxidation by-products in all cases. TOC slowly decreased throughout ozonation, reaching around 40-60% and 60-70% reduction over 90 min, at pH 7 and 3, respectively. PMID- 12088364 TI - Mobility and adsorption capacity of Pb and Zn in a polluted soil from a road environment: laboratory batch experiments. AB - A study of the mobility of lead and zinc in a contaminated soil from a road environment was conducted in order to evaluate the risk of groundwater contamination due to rainwater infiltration. The mobility of trace metals was evaluated using single chemical extractions and single-element sorption experiments. The distribution of trace metallic elements on the various soil fractions investigated with the use of sequential extractions, both before and after single extraction or sorption experiments, enabled the monitoring of changes in their localization. Metals are slightly extracted by sodium chloride solutions. High extraction yield was obtained for lead using EDTA. Lead was removed from the "acid-soluble", "reducible" and "oxidizable" fractions. In contrast, zinc was less dissolved by means of complexation with EDTA, yet it showed very high sensitivity to variations in pH. Following single extractions, metals were redistributed in the "exchangeable" fraction. Sorption experiments evaluated the capacity of the soil to retain additional lead and zinc inputs. The results reveal that pH influences the sorption of these metals and the initial pollution present in the soil may induce desorption phenomena with respect to zinc. The high initial concentrations present in the soil do not seem to prevent the retention of additional metal in significant quantities. Added metals were located in the "exchangeable", "add-soluble" and "reducible" fractions. PMID- 12088365 TI - Monitoring of organochlorine pesticide residues in pikeperch, Stizostedion lucioperca L. in Beysehir Lake (Central Anatolia). AB - Fishes in Beysehir Lake were analysed to determine organochlorine pesticide residues. The more frequently detected compounds are DDT complex (DDT, DDD, DDE and isomers), total HCH complex (alpha-HCH, beta-HCH and gamma-HCH or lindane), aldrin, dieldrin, endrin and heptachlorine. Eighty five percent of the fish samples were found to be contaminated. While 75% of the total samples were contaminated by one or more HCH isomers, 63% of the organochlorinated pesticide is DDT and its metabolites. Aldrin, dieldrin and endrin are minor pesticides in samples. Heptachlorine was found in only one sample. Despite their presence high concentrations are sporadic, the mean values being less than FAO/WHO maximum recommended limits. PMID- 12088366 TI - Reduction of ammonia and odour emissions from pig slurry under slats using oil and foam layers. AB - In Europe, new legislation on environmental protection requires reductions in emissions of ammonia and odour from animal production. Animal housing is identified as one of the biggest ammonia emission sources in pig production, but abatement techniques for animal houses often require high initial cost. The aims of this study were to investigate the use of layers of oil and foam on slurry under slats. It was found that relatively low-cost, used frying oil (mainly rapeseed oil) could reduce ammonia and odour emissions by about 50% in weaner accommodation. The cost for this technique was Pound Sterling 2.60 per pig place per year. Foam showed poor ammonia abatement potential. The oil was biodegradable, so increasing methane emissions from stored pig slurry by about 60%. PMID- 12088367 TI - Removal of sulfacid brilliant pink from an aqueous stream by adsorption onto surfactant-modified Ti-pillared montmorillonite. AB - A modified hydrophobic and organophilic pillared clay (CTAC-TiH-montm) was prepared by exchanging some Ti polymeric cations into the interlamellar space of one Algerian montmorillonite (montm) and then by co-adsorption of some surfactant molecules such as cetyltrimethylammonium chloride (CTAC). These new materials were used in adsorption of an anionic textile dye: Sulfacid brilliant pink (SAP). According to adsorption isotherms, the organic modification of Ti-montmorillonite clay by CTAC surfactant increases the amount of textile dye fixed to more than 1000 mg g(-1). The adsorption experiments showed that a ratio of 3 mmol of CTAC per g of clay and an acidic medium (pH = 4) were the optimal parameters necessary to obtain good adsorption uptake and colourless treated solutions. A comparative study proved the high adsorption capacity of the synthesised adsorbents; they can thus be considered as powerful competitors to activated carbon in the treatment of aqueous textile plants and industry effluents. PMID- 12088368 TI - Biofilm loss in anaerobic immobilized fixed bed reactor system. AB - An anaerobic immobilized fixed bed bioreactor system was studied for treatment of herbal-based pharmaceutical wastewater. The biofilm loss rate (R(S)), shear stress (sigma), biofilm thickness (Lf) and biomass concentration (X,) were investigated for various flow rates applied to the bioreactor during the experimental studies. Some relationships applicable to a fixed bed reactor were obtained between biofilm loss rate, shear stress, biofilm thickness and specific flow rate (Q) with good accuracy. It was evident that shear stress and biofilm loss rate increased with increase in biofilm thickness. A similar trend was also observed with an increase in flow velocity. It was found that shear stress plays a major role in the biofilm detachment mechanism. The biofilm thickness determined in the present investigation was in the range of 0.5 mm to 5.5 mm, and the biofilm loss rate was estimated in the range of 1.7 x 10(-4) to 7.8 x 10(-4) kg COD m(-2) d(-1) at flow rate ranging between 5.0 and 50 1 d(-1). PMID- 12088369 TI - Trace metal sorption ability of insolubilized humic acids. AB - The objective of this work was to study the trace metal sorption ability of Insolubilized Humic Acids (IHA) in comparison with regular Humic Acids (HA). IHA were prepared from HA by heating at 330 degrees C for one hour under a nitrogen atmosphere. A solubility study was carried out at different pH values in NaNO3 (0.1 M) as a background electrolyte. It showed that IHA were insoluble until pH 10 whereas the solubility of HA increased with pH in one main step in the pH range 4-5 over which the soluble fraction of humic acids increased from 10% to 90%. We observed that trace metal sorption was dramatically influenced by these solubility properties. Indeed this study showed that above pH 5-6, HA and IHA may have opposite effects in terms of trace metal sorption. Solubilization of HA made possible the formation of soluble metal-humic complexes, retaining metal in solution even for pH at which hydroxide can form. Under such pH conditions, which very numerous soils show, HA could be involved in the transport and uptake of trace metals in the plant-soil system. In terms of remediation, HA are not a suitable binding phase. On the contrary IHA were not solubilized at neutral and alkaline pH, avoiding then the solubilization of sorbed trace metals. They appeared to be a suitable binding additive phase for future remediation trials. PMID- 12088370 TI - Reduced ammonia emissions from slurry after self-acidification with organic supplements. AB - Ammonia volatilisation from field applied slurries causes environmental hazards and loss of fertilizer value. Acidification of slurry, usually with inorganic or organic acids has previously been used to reduce NH3 emissions. In this study, we present an alternative technique for the acidification of slurry, namely the use of fermentation by endogenous microbes to form organic acids from readily degradable organic compounds. In laboratory experiments, the addition of different sugars (sucrose in dosages of 0.003, 0.01, 0.03, 0.1 and 0.3 mol l(-1), glucose in dosages of 0.05 and 0.1 mol l(-1)) and organic residues (sugar beet residues in dosages of 33 and 330 g fresh weight l(-1), biowaste at 50 g fresh weight l(-1)) to cattle slurry resulted in a considerable decrease in pH, with a minimum pH of 4.7. A subsequent pH increase indicated that the organic acids were probably further degraded with a resultant loss of acidity in the slurry. In a field study, the NH3 emissions from untreated and acidified (pH = 6) slurries were compared after field application (20 m3 ha(-1)). During the first 20 hours, the acidified slurry showed NH3 emissions of less than 5% of the applied ammonia compared to a 26% loss from the untreated slurry. The total emissions of NH4+-N were 32% for acidified and 54% for untreated slurry. Easily degradable organic amendments therefore have the potential to effectively reduce NH3 emissions from slurries and may be an alternative for the use of acids. PMID- 12088371 TI - Biodegradation of gaseous benzene with microbial consortium in a biofilter. AB - The objective of this study was to investigate the biodegradation of gaseous benzene in a biofilter inoculated with benzene-oxidizing microorganisms. The biofilter performance was monitored in terms of benzene removal efficiency and carbon dioxide production. The biofilter was capable of achieving as much as 96% benzene removal efficiency at a residence time of 2 min and an inlet concentration of 220 ppm. During operation with an inlet benzene of 220 ppm, the maximum elimination capacity of the biofilter was 483 g of C6H6 m(-3) day(-1). Under the same conditions, carbon dioxide with a concentration of up to 726 ppm was produced. It was found that carbon dioxide wasproduced at a rate of 608 mg day(-1), which corresponded to a volume of 0.35 l day(-1). Observable features of the microorganisms, meaning microbial activity occurrence in the biofilter, were investigated with the microscopy analysis. PMID- 12088372 TI - Semi-continuous anaerobic co-digestion of agro-wastes. AB - Anaerobic digestion is a well established process for treating many types of organic waste, both solid and liquid. As such, the digestion of cattle slurries and of a range of agricultural wastes has been evaluated and has been successful. Previous batch studies have shown that multi-component mixtures of agro-wastes can be digested successfully, although with some mixtures a degree of both synergism and antagonism occurred. A continuously stirred tank reactor (15 l) was used as a mesophilic (35 degrees C) anaerobic reactor to examine the effect of treating mixtures of fruit and vegetable wastes, cattle slurry, poultry waste and waste activated sludge in which the proportions of the latter two components only were varied. The retention time was kept at 20 days and the loading rate maintained in the range 1.51-1.89 kg VS m(-3) d(-1). Increasing the proportion of poultry waste from 0% to 100% improved the methane yield from 0.164 to 0.244 m CH4 kg(-1) VS added, and caused the VS reduction to decrease slightly. There did not appear to be any synergism or antagonism in the mixtures examined. PMID- 12088373 TI - Purification of pulp and paper mill effluent using Eichornia crassipes. AB - Konark Pulp and Paper Industries Private Limited is a medium size industry producing 1600 m3 of wastewater a day. The existing water treatment system of the industry was found to be ineffective both in performance and economy. In the present study, a new system of treatment has been developed using water hyacinth Eichornia crassipes, coagulation by lime and alum, followed by rapid sand filtration. The performance efficiency of each unit viz. Eichornia treatment; coagulation with lime, with alum, and with lime:alum combinations, and filtration was studied. Water quality parameters considered were Biological Oxygen Demand, Chemical Oxygen Demand, Dissolve Oxygen, Total Dissolved Solids, turbidity, percentage transmission, and water colour. Based on the individual performance of each unit, a continuous system has been designed and was tested. The new system of treatment could treat the wastewater to the discharge standards and also was found economically feasible. Testing culture of fish (tilapia) proved that the treated water could be safely discharged into natural waters. All fish tested, survived and remained healthy throughout the period of testing. Culture of fish further improved the water quality. PMID- 12088374 TI - Use of biological activated carbon to treat mixed gas of toluene and benzene in biofilter. AB - The biodegradation of toluene and benzene in a biofilter filled with cylindrical activated carbon was studied. Three various gaseous flow rates, i.e. 0.25, 0.50 and 0.75 m3 h(-1), corresponding to empty bed gas residences of 75, 37.5 and 25 s, respectively, and total organic load lower than 400 g m(-1) h(-1) were tested. The biofilter proved to be highly efficient in biodegradations of toluene and benzene, and toluene was more easily degraded than benzene. When each inlet load of toluene and benzenewas lower than 150 g m(-3) h(-1), removal rate increased with inlet loads and reached maximum values of 150 and 120 g m(-3) h(-1) for toluene and benzene, respectively. For inlet load higher than the maximum removal capacity conditions, the removal rate decreased with inlet load. The carbon dioxide concentration profile through the biofilter revealed that the mass ratios of carbon dioxide produced to the toluene and benzene removed were 2.15 g CO2 g( 1) toluene and 1.67 g CO2 g(-1) benzene. Model predictions for toluene, benzene and carbon dioxide concentration gradient profiles were in agreement with experimental data for the tested conditions. The observation of biotic community demonstrated that the microbes consisted of bacillus, spore bacillus and fungi, of them spore baxillus was dominant. PMID- 12088375 TI - Modeling a full-scale primary sedimentation tank using artificial neural networks. AB - Modeling the performance of full-scale primary sedimentation tanks has been commonly done using regression-based models, which are empirical relationships derived strictly from observed daily average influent and effluent data. Another approach to model a sedimentation tank is using a hydraulic efficiency model that utilizes tracer studies to characterize the performance of model sedimentation tanks based on eddy diffusion. However, the use of hydraulic efficiency models to predict the dynamic behavior of a full-scale sedimentation tank is very difficult as the development of such models has been done using controlled studies of model tanks. In this paper, another type of model, namely artificial neural network modeling approach, is used to predict the dynamic response of a full-scale primary sedimentation tank. The neuralmodel consists of two separate networks, one uses flow and influent total suspended solids data in order to predict the effluent total suspended solids from the tank, and the other makes predictions of the effluent chemical oxygen demand using data of the flow and influent chemical oxygen demand as inputs. An extensive sampling program was conducted in order to collect a data set to be used in training and validating the networks. A systematic approach was used in the building process of the model which allowed the identification of a parsimonious neural model that is able to learn (and not memorize) from past data and generalize very well to unseen data that were used to validate the model. Theresults seem very promising. The potential of using the model as part of a real-time process control system isalso discussed. PMID- 12088376 TI - Food chain transfer of zinc within the ecosystems of old and modern metalliferous mine wastes. AB - Concentrations of zinc in mine waste, vegetation, invertebrate and small mammals from abandoned mines and a modern mine site have been determined in order to assess and compare environmental risk. Very high concentrations of zinc in mine waste from all sites were reflected in vegetation zinc levels but not in invertebrates and mammals. Physiological regulatory control mechanisms operate to optimise zinc retention, and maintain homeostasis in animals by enhancing absorption of zinc from diets low in the trace element, and by buffering the potential accumulative effects of high dietary levels of the metal. Zinc is not considered to be an acute toxicity factor precluding colonisation of these mine sites by invertebrates and mammals though it maybe a selective force influencing the species composition and relative abundance in animal communities. PMID- 12088377 TI - Measuring and modelling VOC biotransformation rates. AB - A new protocol was used to determine biotransformation rate coefficients for volatile organic compounds in activated sludge systems. The values obtained from the protocol were compared to biotransformation rates obtained from a pilot plant. It was seen that biotransformation rates were easier to obtain from the protocol. Observations also showed that acclimation of the microorganisms to the volatile organic compounds did not change the biotransformation rates. The experimental conditions were then modelled using TOXCHEM+, a mechanistic model to predict volatile organic compound behaviour. The TOXCHEM+ predictions, using system default biotransformation rates, compared favourably with the low biotransformation rates measured by the protocol for bromoform and the chlorinated compounds, while under predicting the biotransformation rates for the non-chlorinated volatile organic compounds. PMID- 12088378 TI - Removal of dissolved organic matter from water by adsorption-flocculation using organobentonite. AB - Bentonite modified with short chain cationic surfactant might be the basis of a new approach to removing dissolved organic matter (DOM) from water. The treatment process involves dispersing bentonite to the DOM-containing water and then adding a small cationic surfactant to result in flocs which are agglomerates of organobentonite and bound DOM. The flocs are then removed from the solution by sedimentation. Experimental results indicate that Benzyltrimethylammonium bromide (BTMA)-bentonite displays a high affinity for DOM, most likely due to aromatic groups in the DOM molecule interacting with the benzene ring in the BTMA ion through increased pi-pi type interactions. Under appropriate operating conditions, 75% DOM removal and nearly 100% bentonite recovery could be achieved by the adsorption-flocculation process using BTMA-bentonite. Additionally, the rapid adsorption kinetics made adsorption-flocculation with BTMA-bentonite attractive for continuous treatment of large volumes of raw water. The bentonite may function as a recyclable surfactant medium for the adsorption and subsequent combustion of organic pollutants. PMID- 12088379 TI - A study of methods to reduce groundwater contamination around the Kimpo landfill in Korea. AB - Four different methods were studied to reduce the groundwater contamination around the Kimpo landfill in Korea by leakage of a leachate. The hydraulic parameters were measured for this study and then the groundwater flow and contaminant transport systems around the Kimpo landfill were simulated with MODFLOW and MT3D models. Both the method of establishing 5 sets of Radial Collector Well Laterals (RCWLs) within the landfill and a method utilizing the wells for dewatering condensed water brought about a 70% leachate elimination efficiency. Their construction costs were cheap, and the maximum concentrations of chloride and phenol in the leachate effluent were high. Meanwhile, a method establishing an interception wall to the 2nd layer at the cirumference of the landfill and 22 sets of RCWLs within it brought about a 92% leachate elimination efficiency. Its construction cost was expensive, and the maximum concentrations of chloride and phenol in the leachate effluent were low. A method of establishing an interception wall to the 3rd layer and 40 sets of RCWLs brought about a 97% leachate elimination efficiency. Its construction cost was the most expensive, and the maximum concentrations of chloride and phenol were low. Conclusively, a method of establishing 22 sets of RCWLs was evaluated as the most stable and economical option to reduce groundwater contamination concentration below drinking water standard. PMID- 12088380 TI - Evaluation of a leaching process coupled with regeneration/recycling of the extractant for treatment of heavy metal contaminated solids. AB - A hydrometallurgical circuit process combining the leaching of metals with the electrolytic regeneration of the mass separating agent is an option to effectively treat heavy metal contaminated solids. This process aims towards a dosed loop utilisation of the extractant by its regeneration and recycling, enabling the use of high cost extractants and the recovery of the heavy metals as well. The circuit process, in which each cycle consists of an extraction step followed by the electrolytic regeneration of the mass separating agent and a second extraction to finally recycle the separating agent and start a new cycle, was applied on a lead contaminated soil from an accumulator manufacturing facility and for the removal of copper from a municipal solid waste incinerator bottom ash. For the first case a 0.13 M citrate solution was used at a pH of 5; for the second 10%; sulphuric acid (pH=0.5). The results showed that above 80% of lead and copper were removed from soil and ash, respectively, when the concentration of mass separating agent was controlled and kept constant. The extraction efficiency of metals from the solid materials was in the same range as the one obtained by two extaction steps using fresh extracting agent. However, due to losses of the mass separating agent the operation in a complete closed loop configuration was not possible. Nevertheless, a substantial reduction in separating agent consumption was achieved. The process proved to be suitable for treating highly contaminated materials, for which the operation parameters of extraction and regeneration are more favourable, also increasing the recovery of metals. PMID- 12088381 TI - Selective removal of chromates by macroporous exchanger Amberlyst A-21. AB - Sorption studies of chromate on weakly basic macroporous anion exchanger Amberlyst A-21 were performed as a function of pH (2 - 9), temperature (20 - 40 degrees C) and concentration (0.4 - 80 mmol.l(-1)). Chromate sorption was observed to decrease with the increase in initial pH of the solution, with a sorption maxima at pH 2 and increased with the increase in temperature. The changes in pH and in the ratio of choride ions released to chromate ions sorbed suggested that the anion exchange process was accomanied by hydrolysis and Donnan invasion. The FTIR spectra and isoeteric heat of sorption besides showing the hydrolysis / Donnan invasion of resin under different experimental conditions of concentration, pH and temperature also showed the dimerization of the chromate species inside the resin phase. PMID- 12088382 TI - Bacterial population of a two-phase anaerobic digestion process treating effluent of cassava starch factory. AB - Different groups of microorganisms in a two-phase anaerobic system were enumerated to evaluate the prevalence of specific groups and species. Total and fecal coliforms showed similar values both in acidogenic and methanogenic reactors. The fecal streptococci were 4-fold higher in the acidogenic reactor, when compared with those of the methanogenic reactor. As expected, no methane forming or sulfate reducing bacteria were found in the acidogenic reactor. The populations of methanogenic bacteria were dominated by a mixed population of straight to curved rods and multicellular filaments which strongly resembled members of the genus Methanosaeta. Seven prevalent species of facultative anaerobic gram-negative bacteria were identified as Previdencia alcalifacienciens, Providencia rettgeri, Enterobacter cloaceae, Citrobacter freundii, Klebsiella oxytoca, Proteus pennri and Yersinia enterocolitica. The species Pseudomonas aeruginosa, Stentophomonas maltophila and Acinotobacter iwoffi, were the most frequently isolated glucose non-fermenting gram-negative bacilli. Among these species, only P. aeruginosa was present in high number in each sample. PMID- 12088383 TI - Tactile perception and manual dexterity in computer users. AB - Recent studies suggest that sensory input generated during highly repetitive tasks can degrade the sensory representation of the hand and eventually lead to sensory and motor problems. In this study, we investigated whether early changes in tactile perception and manual dexterity could be detected in persons exposed to computer tasks. Performance in tests designed to assess tactile perception (grating orientation task for spatial acuity and roughness discrimination) and manual dexterity (grooved pegboard test) was compared between two groups of healthy individuals, matched for age, gender, and experience, who differed in terms of computer habits. One group consisted of frequent users (FU, > 2 h/day, n = 36) and the other of non or occasional users (OU, < 2 h/day, n = 28). Comparison of performance between groups with subjects sorted by gender revealed significant differences (t-test, p < 0.05) in female, but not male, participants. Grating resolution thresholds at the tip on the second and fifth digits were, on average, 40% higher in female FU (n = 13) than in female OU (n = 10) and performance scores on the dexterity test were significantly higher for the left hand. The results of this study indicate that early signs of deterioration in hand function can be present in persons constantly exposed to computer tasks and that these signs are more readily apparent in women than in men. The loss of tactile spatial acuity found in female FU possibly reflect an early consequence of the degraded sensory representation of the hand resulting from constant repetitions of fine motor tasks. PMID- 12088384 TI - Comparison of responses to tensile and compressive stimuli in C-mechanosensitive nociceptors in rat hairy skin. AB - Mechanosensitive nociceptors with unmyelinated axons (C-fibers) were studied in a preparation of isolated skin and nerve from rat. Afferent discharges were recorded while the skin was mechanically stimulated using quantitative stretch (tension) and indentation (compression). The apparatus allowed for generating stimuli of equal magnitudes in both tension and compression. Stimulus-response functions were obtained for individual afferents relating discharge rate to tensile stress or compressive stress. A response threshold and maximal slope were obtained from each function. Thresholds did not differ significantly for compression and tension nor did the maximal slopes. We conclude that C nociceptors are equally sensitive to tensile and compressive stress. PMID- 12088385 TI - A four-channel analysis of the tactile sensitivity of the fingertip: frequency selectivity, spatial summation, and temporal summation. AB - Thresholds were measured for the detection of vibratory stimuli of variable frequency and duration applied to the index fingertip and thenar eminence through contactors of different sizes. The effects of stimulus frequency could be accounted for by the frequency characteristics of the Pacinian (P), non-Pacinian (NP) I, and NP III channels previously determined for the thenar eminence (Bolanowski et al., J Acoust Soc Am 84: 1680-1694, 1988; Gescheider et al., Somatosens Mot Res 18: 191-201, 2001). The effect of changing stimulus duration was also essentially identical for both sites, demonstrating the same amount of temporal summation in the P channel. Although the effect of changing stimulus frequency and changing stimulus duration did not differ for the two sites, the effect of varying the size of the stimulus was significantly greater for the thenar eminence than for the fingertip. The attenuated amount of spatial summation on the fingertip was interpreted as an indication that the mechanism of spatial summation consists of the operations of both neural integration and probability summation. PMID- 12088387 TI - Synthetic heat at mild temperatures. AB - "Synthetic heat", also known as the heat grill illusion, occurs when contact with spatially adjacent warm and cold stimuli produce a sensation of "heat". This phenomenon has been explained as a painful perception that occurs when warm stimulation inhibits cold-sensitive neurons in the spinothalamic tract (STT), which in turn unmasks activity in the pain pathway caused by stimulation of C polymodal nociceptors (CPNs). The "unmasking model" was tested in experiment 1 by combining warm (35-40 degrees C) and cool (> or = 27 degrees C) stimuli that were too mild to stimulate CPNs. After discovering that these temperatures produced nonpainful heat, experiment 2 was designed to determine whether heat could be induced when near-threshold cooling was paired with mild warmth, and whether lowering the base temperature for cooling would increase the noxious (burning, stinging) components of heat for fixed cooling steps of 1-3 degrees C. Cooling by just 1 degrees C from a base temperature of 33 degrees C led to reports of heat on more than 1/3 of trials, and cooling by just 3 degrees C evoked heat on 75% of trials. Lowering the base temperature to 31 or 29 degrees C increased reports of heat and burning but did not produce significant reports of pain. Perception of nonpainful heat at such mild temperatures indicates either that cold-sensitive nociceptors with thresholds very similar to cold fibers innervate hairy skin in humans, or that heat can result from integration of warm fiber and cold fiber activity, perhaps via convergence on nonspecific (e.g., WDR) neurons in the STT. PMID- 12088386 TI - Effects of alterations of the vibrissae-related organization of thalamocortical axons upon the organization and outgrowth of intracortical connections in the barrelfield of the rat. AB - Previous studies have shown that intracortical projections in layer IV of the vibrissae representation of primary somatosensory cortex (S-I) are arrayed in a pattern complementary to that of thalamocortical axons (TCAs). Elevation of cortical serotonin (5-HT) in rats during the first postnatal week results in a transient disruption of the vibrissae-related pattern of TCAs and layer IV neurons in S-I. The present study examines the influence of elevated cortical 5 HT levels and the attendant loss of vibrissae-related TCA clusters on the organization of S-I intracortical connections. Cortical 5-HT was elevated in neonatal rats via chronic injections of clorgyline from birth until P-6. Animals were euthanized on P-6 or allowed to survive an additional 4 days without further clorgyline treatment. Distributions of TCAs and intracortical axons were assessed via application of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (Di-I) and 4-(4-(dihexadecylamino)styryl)-N-methylpyridinium iodide (Di-A) to the thalamic radiations and directly into the cortical barrelfield, respectively. Chronic administration of clorgyline resulted in a loss of the vibrissae-related organization of TCAs in layer IV of S-I. There was also a loss of the complementary pattern of intracortical projections in layer IV of this region. Discontinuation of clorgyline treatment resulted in a return of the vibrissae-related pattern of TCAs as well as the complementary pattern of intracortical projections. These results are consistent with the conclusion that the normal organization of intracortical projections in this region of S-I depends on the presence of the orderly array of TCAs. PMID- 12088389 TI - Decreased cutaneous sensory axon-reflex vasodilatation below the lesion in patients with complete spinal cord injury. AB - The histamine-induced skin flare response has been considered of practical value in determining the level of a spinal cord lesion, but clinical observations have varied widely with regard to the nature and degree of change below the lesion. We have quantified cutaneous sensory axon-reflex vasodilatation in patients with complete spinal cord injury (SCI) above and below the lesion, and compared the findings with normal subjects. Axon-reflex vasodilatation was induced by intradermal histamine injection, and measured by (a) laser Doppler fluxmetry and (b) tracing the surface area of the flare. Axon-reflex vasodilatation was present in all SCI patients above and below the lesion, but was significantly diminished below the lesion by both measures (pflux rise = 0.0008; pflare = 0.023), and in comparison with controls (by 39%). The flux increase was significantly correlated with the area of flare (r = 0.82; p = 0.02). Axon-reflex vasodilatation and visual analogue scale (VAS) pain scores on histamine injection were not significantly different above the lesion in SCI patients from controls. Baseline laser Doppler flux was not different at any test site in SCI and normal subjects. The cutaneous sensory axon-reflex is thus significantly diminished in SCI patients below the level of the lesion, but the underlying mechanism is unclear. A possible explanation under investigation is that increased basal or reflex sympathetic vasoconstriction mediated via the isolated spinal cord may counteract the vasodilatation produced by the cutaneous sensory terminals. PMID- 12088388 TI - The relationship between MI and SMA afferents and cerebellar and pallidal efferents in the macaque monkey. AB - The purpose of the present study was to determine the interrelationship between the thalamic afferents arising from the cerebellum (Cb) and the internal segment of the globus pallidus (GPi) with the neurons projecting to the primary motor cortex (MI) and to the supplementary motor area (SMA). We combined fluorescent retrograde tracers with a double anterograde labeling technique. Multiple injections of a combination of Diamidino Yellow and Fast Blue were made into either the MI or SMA hand/arm representation as determined by intracortical microstimulation. In the same animal, biotinylated dextran amine was injected into the GPi and horseradish peroxidase conjugated to wheat germ agglutinin was injected into the contralateral cerebellar nuclei. The results revealed that the cerebellar and pallidal thalamic territories are largely separate. The ventral anterior nucleus (VA) and the ventral lateral nucleus pars oralis (VLo) contained a greater density of pallidal labeling while a greater density of cerebellar label was observed more caudally in the ventral posterior lateral nucleus pars oralis (VPLo) as well as in nucleus X (X). Moreover, we observed that the greatest coincidence of retrograde cell labeling was within the pallidal thalamic territory following the SMA injections and within the cerebellar thalamic territory following the MI injections. However, interdigitating foci of pallidal and cerebellar label were also observed particularly in the ventral lateral nucleus pars oralis (VLo) and the ventral lateral nucleus pars caudalis (VLc). In both VLo and VLc, we additionally observed coincidence between the cerebellar labeling and SMA projection neurons as well as between pallidal labeling and MI projection neurons. These data suggest that while MI primarily receives inputs originating from Cb and SMA primarily receives inputs originating from GPi, it also appears that MI and SMA receive secondary afferents arising from GPi and Cb, respectively. PMID- 12088390 TI - The effects of long-standing limb loss on anatomical reorganization of the somatosensory afferents in the brainstem and spinal cord. AB - We examined the terminations of sensory afferents in the brainstem and spinal cord of squirrel monkeys and prosimian galagos 4-8 years after a therapeutic forelimb or hindlimb amputation within 2 months of birth. In each animal, the distributions of labeled sensory afferent terminations from remaining body parts proximal to the limb stump were much more extensive than in normal animals. These sprouted afferents extended into the portions of the dorsal horn of the spinal cord as well as the cuneate and external cuneate nuclei of the brainstem (forelimb amputees) or spinal Clarke's column (hindlimb amputee) related to the amputated limb. Such reorganization in sensory afferents along with reorganization of the motor efferents to muscles (Wu and Kaas, J Neurosci 19: 7679-7697, 1999, Neuron 28: 967-978, 2000) may provide a basis for mislocated phantom sensations of missing forelimb movements accompanying actual shoulder movements during cortical stimulation or movement imagery in patients with amputations. PMID- 12088391 TI - Environmental thermal stress. AB - Thermal stress from cold and heat can affect health and productivity in a wide range of environmental and workload conditions. Health risks typically occur in the outer zones of heat and cold stress, but are also related to workload. Environmental factors related to thermal stress are reviewed. Individuals undergo thermoregulatory physiologic changes to adapt and these changes are reviewed. Heat and cold related illnesses are reviewed as well as their appropriate therapy. Published standards, thresholds and recommendations regarding work practices, personal protection and types of thermal loads are reviewed. PMID- 12088392 TI - Bacterial and fungal aerosols in indoor environment in Central and Eastern European countries. AB - Studies of indoor bioaerosols conducted in Central and Eastern European countries, as a result of the scarcity of funding, mostly do not attain the level presented by similar studies in Northern America and Western Europe. For socio economic reasons, most of the intense studies on indoor bioaerosols in Central and Eastern European countries were carried out in industrial facilities and have contributed significantly to occupational health science. In contrast, until recently, insufficient of studies have been conducted on bioaerosols of residential and communal premises (dwellings, offices, schools, etc.) and no network for monitoring the microbiological quality of air in such premises exists. In Poland, in the mid-1990s complex bioaerosol investigations were carried out by the Bioaerosol Group at the Institute of Occupational Medicine and Environmental Health in Sosnowiec. The concentrations of airborne bacteria and fungi in dwellings without mold problems were between 88-4297 cfu/m(3) and 0-1997 cfu/m(3), while in moldy homes they were 178-4751 cfu/m(3) and 49-16,968 cfu/m(3), respectively. As many as 167 microbial species were isolated from the air of examined dwellings. Most frequently occurred Gram-positive cocci (Micrococcus/ Kocuria spp., Staphylococcus spp.), endospore-forming bacilli (Bacillus spp.), Gram-negative bacteria (Pseudomonadaceae, Aeromonas spp.), filamentous fungi (Penicillium spp., Aspergillus spp.), and yeasts. Notable studies of indoor bioaerosols have also been performed in the other Central and Eastern European countries: Lithuania, Latvia, Estonia, Russian Federation, Ukraine, Czech Republic, Slovakia, Bulgaria and Hungary, are reviewed in this article. The lack of reference limit values for bioaerosols seriously hinders interpretation of results obtained in various countries. The following residential limit values (RLV) for dwellings and communal premises are proposed for the concentration of airborne bacteria, fungi and bacterial endotoxin: 5 x 10(3) cfu/m(3), 5 x 10(3) cfu/m(3) and 5 ng/m(3) (50 EU), respectively. The proposed values of occupational exposure limit (OEL) for industrial settings contaminated by organic dust are 100 x (3) cfu/m(3), 50 x (3) cfu/m(3) and 200 ng/m(3) (2000 EU), respectively. It is also proposed that the presence in indoor air of microorganisms from risk groups 3 and 4 of European Community Directive 2000/54/EC (e.g., Mycobacterium tuberculosis, Bacillus anthracis, Coxiella burnetii), independently of the concentration, should always be inadmissible and result in preventive actions. PMID- 12088393 TI - Coexistence DNA of Borrelia burgdorferi sensu lato and Babesia microti in Ixodes ricinus ticks from north-western Poland. AB - The tick Ixodes ricinus may carry microorganisms which cause serious human and animal diseases, i.a., the Lyme disease (borreliosis), caused by the spirochaete Borrelia burgdorferi sensu lato and babesiosis, induced by the protozoan Babesia microti. Both microbe species may co-occur in the same and other species of the genus tick and produce a mixed infection in humans and animals. The major objective of the study was to identify DNA of B. burgdorferi and B. microti in the I. ricinus ticks collected in spring and autumn 1999 from 6 sites in north western Poland. The microbial DNA was identified with polymerase chain reaction (PCR). The marker used to detect the B. burgdorferi s.l. DNA was a fragment of the fla gene encoding the protein flagellin, while the B. microti DNA was detected with a fragment of the gene encoding 16S rRNA. A total of 550, 1,160, and 385 tick adults, nymphs, and larvae, respectively, were examined. Among the 155 (7.4%) B. burgdorferi- infected ticks and the 130 (6.2%) infected with B. microti, mixed infection was detected in 0.6% of individuals. The prevalence of coinfection differed between the tick developmental stages. Coinfection was most prevalent (3.1%) in females, males and nymphs being less affected (0.4 and 0.2%, respectively). No coinfection was revealed in the tick larvae. The study described was the first of its kind to be conducted in the former District of Szczecin. For the phenomenon of microbial co-occurrence and related mixed infections to be properly evaluated, the research will be continued. PMID- 12088394 TI - The use of the radioisotope method in studies of pesticide penetration into the eyeball. AB - The studies concerning the effects of pesticides on the human body focused mainly on their local action or chronic poisoning of the organism. In this study we examine the pesticide penetration into the eyeball resulting from direct contact with the eye. We used an isotope-labelled carbamate pesticide -- carbaryl. The determinations of the amount and concentration of this substance in the cornea, aqueous humour, vitreous humour and retina were performed using the method measuring beta radiation emitted by radioactive carbon. The results revealed measurable concentrations of labelled carbamate in the cornea and aqueous humour 10 and 30 min after application. The levels of this pesticide in the vitreous humour and retina were very low and difficult to analyse statistically. The described method appears to be useful in determining the range and rate of eye penetration by environmental toxins having direct contact with the eyeball. PMID- 12088395 TI - House dust mites and their allergens in Danish mattresses -- results from a population based study. AB - The purpose of this study was to identify the level of house dust mites (HDMs) and their allergens in mattresses, not selected on their owners atopic status, and to find associated factors. Dust was collected from 68 mattresses. The recruitment was population-based and conducted during the screening phase of a HDM intervention study. The visited persons declared to have had a "cold" bedroom the previous winter. HDMs were counted and dust was analysed by ELISA for Der 1 (= Der f 1+ Der p 1+ Der m 1). Multiple regression analysis was carried out to find housing conditions associated with high HDM levels. Type of housing, mattress age and self-assessed winter bedroom-temperature explained 47% of Der 1. Median concentrations were 3.77 microg Der 1/g and 1 HDM/0.1 g dust. Both immunochemically and microscopically Dermatophagoides farinae was dominant; D. pteronyssinus less frequent but important; and D. microceras insignificant. In 62% of these suburban homes the mattress dust exceeded 2 microg Der 1/g; and measurement of both Der f 1 and Der p 1 was necessary and sufficient to evaluate HDM allergen exposure. The association with a high HDM level was highest and most consistent for one-family houses. PMID- 12088396 TI - Air contaminants in different European farming environments. AB - Farmers are known to be at high risk from the development of occupational airway disease. The first stage of the European farmers' study has shown that pig farmers in Denmark and Germany, poultry farmers in Switzerland and greenhouse workers in Spain were at highest risk for work-related respiratory symptoms. Therefore, the aim of this study was to determine exposure levels at relevant farm workplaces. Dust and endotoxin levels as well as microbiological concentrations were determined in 213 crop and animal farming environments by personal sampling. The highest total dust concentrations were found in poultry houses in Switzerland with median concentrations of 7.01 mg/m(3). The median airborne endotoxin concentrations in total dust ranged between 0.36 ng/m(3) in Spanish greenhouses and 257.58 ng/m(3) in poultry houses in Switzerland. Likewise, the highest median concentrations of total (2.0 x (7) cells/m(3)) and active fungi (4.4 x (5) cfu/m(3)) have been found in Swiss poultry houses. The predominant fungus taxa discovered in poultry houses were Eurotium spp. and thermophilic fungi. Cladosporium and Botrytis were mainly detected in greenhouses. The exposure level found in this study might put the farmers at risk from respiratory diseases. PMID- 12088398 TI - Detection of Borrelia burgdorferi sensu lato in mosquitoes (Culicidae) in recreational areas of the city of Szczecin. AB - The mosquitoes were caught within the period of their highest activity, i.e. June August 2001, in recreational areas of the city of Szczecin. Spirochetes, Borrelia burgdorferi sensu lato were detected in mosquitoes with the aid of the method of indirect immunofluorescence (IFA) using rabbit anti-Borrelia burgdorferi antibodies and goat anti-rabbit IgG marked with fluorescein isothiocyanate (FITC). A total of 639 mosquito females representing genera Aedes (99.1%) and Culex (0.9%) were collected. The mean value of the infection rate of mosquitoes from the area studied was 1.25%. The highest infection rate was recorded in June (3.2%), while the lowest -- in July (0.6%). All mosquitoes infected with Borrelia burgdorferi spirochetes belonged to the genus Aedes. The results of the present study confirm a potential role of those arthropods in epidemiology of Lyme borreliosis PMID- 12088397 TI - Second inter-laboratory study comparing endotoxin assay results from cotton dust. AB - Previously, a large two-part inter-laboratory round robin endotoxin assay study was completed. This first study showed that when cotton dust samples, which are practically identical, are assayed for endotoxin that the intra- laboratory results had a very small variation while intra-laboratory results of the sample had a very high variation. In the first part of the study, each laboratory followed its own in-house assay protocol; but in the second part of the study, when the extraction protocol was standardized, the inter-laboratory results showed a lower variation, which suggested that with further standardization, further reduction of differences between laboratories might be achieved in order that results between laboratories would become more comparable. The results stimulated interest in extending the study to include cotton dust with two levels of endotoxin, standardization of the extraction protocol, and using the same assay kit from the same production lot. The results of this second round robin endotoxin assay study indicate that differences between laboratories are still high, but most of the laboratories could discern the cotton dusts with the different levels of endotoxin. PMID- 12088399 TI - Indoor fungi and their ciliostatic metabolites. AB - According to epidemiological studies, it is possible that some secondary metabolites of indoor airborne fungi could be responsible for health troubles which occupants suffer from. In our previous experiments, a model with tracheal rings of 1-day-old chicks in vitro was shown to be a very suitable method to study the ciliostatic chloroform-extractable endo- and/or exometabolites of filamentous fungi. In this study we isolated the filamentous fungi from walls of "mouldy" dwellings and schools (cultivation on dichloran 18% glycerol agar at 25 and 37 degrees C for 10 d) in Slovakia. We studied the ciliostatic effect of the chloroform-extractable endo- and exometabolites of 96 representative isolates (stationary cultivation on the liquid medium with 2% of yeast extract and 10% of sucrose at 25 degrees C for 10 days) on the cilia movement in tracheal organ cultures of 1-day-old chickens in vitro after 24, 48 and 72 hrs (incubation in the minimal essential medium according to Eagle with Earl s salts and 20 microg of extract of metabolites dissolved in dimethylsulfoxide per 1 mL). Strains of Penicillium Link: Fr. sp., Aspergillus versicolor (Vuill.) Tiraboschi, A. flavus Link, Cladosporium sphaerospermum Penzig and C. cladosporioides (Fres.) de Vries were isolated most frequently. Two A. flavus isolates were able to produce aflatoxins B1, B2, G1, G2 in vitro after cultivation on the liquid medium with 20% sucrose and 2% yeast extract. This is the first isolation of aflatoxigenic A. flavus strains from dwellings in Slovakia. All frequently isolated strains produced secondary metabolites with the strongest ciliostatic activity -- their exo- and endometabolites stopped tracheal ciliary movement in chicks till 24 h. There are some toxic fungal metabolites in the indoor air not only with the ability to destroy ciliary movement in the upper airways in vitro but, probably, during long-lasting exposure to cause general intoxication of macroorganism via lung tissue. PMID- 12088400 TI - Betula and Populus pollen counts and meteorological conditions in Szczecin, Poland. AB - The aim of the study was to analyse a relationship between the meteorological conditions and the pollen counts of selected allergenic taxa (Betula spp., Populus spp.) in the air of Szczecin city (north western Poland) in 2001. Apart from the individual rhythm of plant pollination, weather conditions are considered the most important factors determining the dispersion and content of pollen in the air. The meteorological parameters analysed were the minimum, maximum and average values of air temperature, relative humidity, solar radiation and wind speed. The beginning and end of a season were established by the 98 % method. The concentration of birch pollen in the air in 2001 was very high, the pollen season started in the third decade of April and lasted till the 10 May. The highest airborne concentration of 3,712 grains in 1 m(3) per 24 h was noted at the beginning of May on a sunny day with strong wind, and air temperature above 20 degrees C. The concentration of poplar pollen in 2001 was low. The pollen season started from the beginning of April and lasted till the beginning of May. The maximum concentration of airborne poplar pollen of 222 grains in 1 m(3) per 24 h was observed in the third decade of April and was preceded by several days with low temperature (1- 2 degrees C), rain, snowfalls and strong wind. A positive and statistically significant correlation was found between the air temperature and the birch pollen concentration, while a similar but negative correlation was found for poplar pollen. PMID- 12088401 TI - Qualitative and quantitative analysis of agricultural dust in working environment. AB - The presented quantitative and qualitative analysis of dust in agricultural working environment is a continuation of the process of recognizing the exposure to dust among private farmers. The study covers the following: determination of respirable fraction of dust in the respiratory zone (on the background of total dust) while performing individual farming activities which constitute an annual work cycle, organic and mineral components of settled dust for basic groups of farming activities, and the main mineral pathogenic component - free silica in airborne and settled dust. The study was conducted on 5 farms specialising in: cultivation of cereals, root plants, vegetables, dairy cattle and swine breeding and mixed production. The analysis of settled dust covered 17 types of dust accompanying field work and farm/indoor activities. Studies of airborne dust were conducted on farmers while performing 40 main work activities which contributed to the annual work cycle. Results of the study confirmed the following: agricultural work activities are accompanied by a high level of dustiness and showed the presence of a respirable fraction in airborne dust of up to 25%, a higher level of pathogenic free silica SiO(2) in settled dust samples in the working environment of a farmer, compared to dust in respiratory zone, a comparable level of SiO(2) in total and respirable airborne dust, and a high level of organic component in settled dust at work activities with plant material. These results indicated that the evaluation of farmers' exposure to dust should be based on the examination of samples taken in the respiratory zone while performing individual work activities PMID- 12088402 TI - Selected socio-economic features and the prevalence of peptic ulcer among Polish rural population. AB - The aim of the study was to determine the relationship between the prevalence of peptic ulcer and the occurrence of selected socio-economic features among Polish rural population. The study was conducted based on the all- Polish representative study of the state of health of rural population, and covered a group of 6,512 rural inhabitants aged 20-64 -- 3,107 males and 3,405 females selected by two stage stratified sampling. Peptic ulcer was diagnosed in 348 people in the study (5.3%): 250 males (8.0%) and 98 females (2.9%). Duodenal ulcer occurred in 3.2% of people examined, followed by gastric ulcer -- 1.2%, duodenal and gastric ulcer -- 0.2%, and 0.9% of patients underwent surgical procedures due to peptic ulcer. Peptic ulcer occurred more frequently among people with a lower education level (lack of education -- 7.8%, elementary school education -- 5.8%), compared to those with higher education categories (elementary vocational -- 4.9%, secondary school and college -- 3.7%). The disease was more often diagnosed among respondents who described their material standard as poor (7.7%), compared to those who described this standard as good (4.0%). Among people who considered their material standard as poor, gastric ulcer was noted more frequently than duodenal ulcer. A correlation was observed between the prevalence of peptic ulcer and such socio-economic features of Polish rural population as the level of education and material standard. PMID- 12088403 TI - Exposure to airborne microorganisms in furniture factories. AB - Microbiological air sampling was performed in 2 furniture factories located in eastern Poland. In one factory furniture were made from fibreboards and chipboards while in the other from beech wood. It was found that the concentration of total microorganisms (bacteria + fungi) in the air of the facility using beech wood for furniture production (mean 10.7 x (3) cfu/m(3), range 3.3 27.5 x (3) cfu/m(3)) was significantly higher (p < 0.01) compared to microbial concentration in the facility using fibre- and chipboards (mean 3.6 x (3) cfu/m(3), range 1.9-6.2 x (3) cfu/m(3)). On average, the commonest microorganisms in the air of the furniture factories were corynebacteria (Corynebacterium spp., Arthrobacter spp., Brevibacterium spp.) which formed 18.1 50.0% of the total airborne microflora, and fungi (mostly Aspergillus spp., Penicillium spp., Absidia spp. and yeasts) which formed 6.2-54.4% of the total count. The values of the respirable fraction of airborne microflora in the furniture factories varied within fairly wide limits and were between 15.0-62.4%. Altogether, 28 species or genera of bacteria and 12 species or genera of fungi were identified in the air of examined factories, of which respectively 8 and 7 species or genera were reported as having allergenic and/or immunotoxic properties. In conclusion, the workers of furniture factories are exposed to relatively low concentrations of airborne microorganisms which do not exceed the suggested occupational exposure limits. Nevertheless, the presence of allergenic and/or immunotoxic microbial species in the air of factories poses a potential risk of respiratory disease, in particular in sensitive workers. PMID- 12088404 TI - Response of furniture factory workers to work-related airborne allergens. AB - The aim of this work was to determine the reactivity of furniture factory workers to microbial allergens associated with wood dust. Allergological examinations by skin and precipitin tests were performed in 48 workers employed in a factory producing furniture from fibreboards and chipboards, and in 32 healthy urban dwellers not exposed to organic dusts (referents). The skin test was performed by the intradermal method with the saline extracts of the cultures of 3 microbial species (Rahnella sp., Arthrobacter globiformis, Aspergillus fumigatus) associated with wood dust. Skin reactions were recorded after 20 minutes, 8 hours and 24 hours and graded 1-4, depending on the diameter of the reaction. The agar gel test for the presence of precipitins in serum was performed with the extracts of 15 microbial isolates. The furniture factory workers showed a high skin response to the extracts of environmental microbes. The frequency of early grade 2 reactions (diameter 10 mm) to the extract of Rahnella sp. was 64.6% among furniture workers, being significantly higher (p < 0.001) compared to reference group (18.7%). High frequencies of grade 2 reactions in furniture workers were also found with the extracts of A. globiformis and A. fumigatus (52.1% and 62.5%, respectively). The frequencies of grade 2 delayed (after 8 h) and late (after 24 h) reactions to Rahnella sp. in furniture workers were non-specifically high (97.9%/93.7%) while the response rates to A. globiformis and A. fumigatus were much lower (10.4%/25.0%, and 4.2%/37.5%, respectively). In agar-gel test for detection of precipitins, in most cases very low percentages of positive reactions (0-2.1%) were noted in furniture factory workers. The only exception was a high percentage of positive reactions (27.1%) to the antigen of Pseudomonas maltophilia, which was significantly greater in furniture workers compared to the reference group (p < 0.01). The obtained results suggest that early allergic reactions to microorganisms associated with wood dust are common among workers of furniture industry, which may increase a potential risk of work-related disease in this occupational group. PMID- 12088406 TI - Study on the occurrence of Borrelia burgdorferi sensu lato and tick-borne encephalitis virus (TBEV) in ticks collected in Lublin region (eastern Poland). AB - In 114 Ixodes ricinus ticks from 1 district of Lublin region (eastern Poland) examined by dark field microscopy method, the presence of motile spirochetes, morphologically corresponding to Borrelia species was detected in 8.8% of the total examined ticks. The highest infection rate was noted among females (16.7%), much lower in males (7.1%) and nymphs (4.0%). Examination of 550 ticks collected from 3 districts of the Lublin region by polymerase chain reaction (PCR) showed that 5.3% of the total number of ticks were infected with Borrelia spirochetes. The highest rates of Borrelia infection were observed in Ixodes ricinus ticks from the Zamosc and Lublin districts (9.6% and 4.7% respectively). In the Wlodawa district, only 2.4% ticks showed the presence of Borrelia DNA. In contrast to the results obtained by dark field microscopy method, the highest infection rate was noted in males (11.2%), followed by females (6.9%) and nymphs (1.7%). 57 Ixodes ricinus ticks collected from 3 Lublin districts (Lubartow, Lublin, Radzyn Podlaski) were tested for the presence of tick-borne encephalitis virus (TBEV) by the inoculation of 5-week-old Swiss mice, followed by blind passages and inoculation of GMK cell culture with brain suspension of the infected mice. One strain of TBEV was isolated from a pool of 24 I. ricinus ticks collected from the Radzyn Podlaski district. The minimum infection rate of ticks from this district with TBEV was estimated as 4.2%, and in the total area of Lublin region estimated as 1.8%. PMID- 12088405 TI - Work-related symptoms among furniture factory workers in Lublin region (eastern Poland). AB - 48 woodworkers employed in the furniture factory were examined. The control group consisted of 41 office workers with no exposure to organic dust. The examination included: interview on work-related symptoms, physical examination, and lung function test performed before and after the working-day. 38 out of 48 (79.2%) woodworkers reported work-related symptoms. The most common complaint was dry cough reported by 25 workers (52.1%), followed by general malaise -- reported by 17 (35.45%), conjunctivitis -- by 16 (33.3%), rhinitis - by 16 (33.3%), and skin symptoms by 16 (33.3%). Other symptoms such as headache, shortness of breath and chest pain occurred less frequently. Subjects working in initial processing and board processing departments had a higher prevalence of cough compared to workers employed in the varnishing department (p < 0.01). The prevalence of skin symptoms was significantly higher in board processing and varnishing departments compared to initial processing department (p < 0.05). Occupational asthma and allergic alveolitis were recorded in 3 out of 48 (6.2%) and 2 out of 48 (4.2%) workers, respectively. Baseline FVC and FEV(1) values were lower in woodworkers compared to controls (p < 0.01). The increased lung function parameters (FVC, FEV(1)) were observed in woodworkers who smoked compared to non-smokers. The difference was not statistically significant. There was a significant over-shift decrease of all measured spirometric values: FVC, FEV(1)), FEV(1)) /VC, PEF among woodworkers (p < 0.001). There was a significant pre-shift, post-shift decline in FVC, FEV(1)), FVC/FEV(1)), and PEF among workers under 30 years of age (p < 0.001). The same tendency was seen for FVC and FEV(1)) in subjects over 30. The percentage changes in FVC and FEV(1)) were greater in the group of younger workers (15.1% and 17.6%) respectively, than in the group of older subject (6.2%, 7.1%). The difference was not statistically significant. PMID- 12088407 TI - Longterm decrease in the CD57 lymphocyte subset in a patient with chronic Lyme disease. AB - Lyme disease is a tickborne illness caused by the spirochete Borrelia burgdorferi. In a previous report we described a decrease in the CD57 lymphocyte subset in patients with chronic Lyme disease. We have now identified a patient with chronic relapsing and remitting symptoms of Lyme disease who had decreased levels of CD57 lymphocytes over 10 years. This observation represents the longest duration of an immunologic abnormality ever documented in chronic Lyme disease. The CD57 lymphocyte subset appears to be a useful marker of longterm infection with the Lyme disease spirochete. PMID- 12088408 TI - Allergic contact urticaria and rhinitis to roe deer (Capreolus capreolus) in a hunter. AB - Roe deer (Capreolus capreolus) is one of the most common game mammals in Europe, where hundreds of thousands people are exposed to this animal. Despite this fact, we are aware of only two cases of allergy to roe deer published until recently, one case of allergic rhinoconjunctivitis and asthma and the second of contact urticaria. We describe another case with co-existing allergic contact urticaria and rhinitis in a 55-year old male professional hunter. The symptoms were provoked only by exposure to roe deer, and there were no other past or present allergic diseases. Specific IgE was found to following animal allergens: cow dander (CAP class 5), goat epithelium and horse dander (each CAP class 4), dog epithelium, dog dander and swine epithelium (each CAP class 2). Skin prick tests have shown positive reaction only to cow epithelium (+). Because of lack of deer dander allergen for specific IgE and skin tests, we have confirmed the causal relationship between exposure to roe deer and allergy using the rub test with roe deer's fur. There was a clearly positive urticarial reaction on the patient's skin accompanied by nasal itch, sneezing and rhinorrhea. No reaction was seen in a control person. We surmise that the positive tests with cow epithelium seen in this patient may result from a cross-reactivity to deer allergens. We conclude that although occupational allergies to roe deer seem to be rare, such possibility should be always considered among people having contact with these animals. PMID- 12088409 TI - B cells of aged mice show decreased expansion in response to antigen, but are normal in effector function. AB - Increased dysfunction of the immune system with age can be attributed to developmental changes in cell types critical for proper immune responses. Previous studies have shown defects in humoral responses of aged individuals, but have not distinguished between aged T-cell/microenvironment and intrinsic B-cell defects. Here adoptive transfer of antigen-specific transgenic B cells compared early immunopoeisis from young and aged donors in a young recipient environment. B cells from aged donors demonstrated decreased antigen-induced expansion, particularly in the lymph nodes; however, they acquired a germinal center phenotype at frequencies similar to B cells from young donors. Additionally, aged B cells produced equivalent levels of antigen-specific antibody that underwent affinity maturation and isotype switching and demonstrated similar numbers of antibody-secreting cells of switched isotype. Thus, the ability of aged B cells to respond appropriately to T-dependent antigens and differentiate into high affinity, isotype-switched, antibody-secreting cells appears to be intact. PMID- 12088410 TI - Observation of antigen-dependent CD8+ T-cell/ dendritic cell interactions in vivo. AB - In order to track hematopoetic cells of all lineages unambiguously at all stages of development, we have developed C57BL/6 mice that express a transgene coding for green fluorescent protein (GFP) under control of the human ubiquitin C promoter. These mice, called UBI-GFP/BL6, express GFP in all tissues examined, with high levels of GFP expression observed in hematopoetic cells. UBI-GFP/BL6 mice are unique in that B cells, T cells, and dendritic cells have distinct levels of GFP fluorescence. In cell transfer experiments, leukocytes from UBI GFP/BL6 mice are readily identified by FACS or fluorescence microscopy. We demonstrate that transplanted UBI-GFP/BL6 dendritic cells are easily identified in secondary lymphoid tissues. Direct interactions between individual dendritic cells and multiple naive CD8+ T cells are observed in lymph nodes within 12 h of cell transfer and require loading of the dendritic cells with the appropriate peptide antigen. Dendritic cells undergo specific morphologic changes following interactions with antigen-specific T cells. PMID- 12088411 TI - Analysis of the individual role of the TCRzeta chain in transgenic mice after conditional activation with chemical inducers of dimerization. AB - Signaling through the TCR/CD3 complex plays a critical role in T-cell development and activation. Gene-targeted mice lacking particular components of this complex show arrested T-cell development in the thymus. As all TCR/CD3 components are required for efficient surface expression of the complex, it is difficult to assess the specific signaling role of each receptor component. To overcome this problem, we designed a strategy to examine the specific role(s) of individual receptor chains. A chimeric protein, containing binding domains for chemical inducers of dimerization fused to the cytoplasmic tail of TCRzeta, was generated. Activation of the chimeric receptor after stimulation with chemical dimerizers in Jurkat cells showed tyrosine phosphorylation of the TCRzeta chain chimera, recruitment of phosphorylated Zap70, and generation of NFAT in a reporter assay. Analysis of thymocytes from transgenic mice expressing this chimeric receptor showed that intracytoplasmic crosslinking of the chimera induced tyrosine phosphorylation of the protein, as well as a slow and very weak calcium mobilization response. However, this signaling did not lead to increased expression of activation markers, T-cell proliferation, or apoptosis. In addition, stimulation of thymocytes in suspension or in fetal thymic organ cultures with chemical inducers of dimerization did not lead to alterations in positive or negative selection. We conclude that signaling through the TCRzeta chain alone is not sufficient to generate downstream events leading to full T cell activation or thymocyte selection; instead, additional CD3 components must be required to induce a functional response in primary thymocytes and peripheral T cells. PMID- 12088412 TI - Antigen presentation of a modified tumor-derived peptide by tumor infiltrating lymphocytes. AB - CD8+ T-lymphocytes recognize peptides in the context of major histocompatibility complex (MHC) class I antigens. Upon activation, these cells differentiate into effector cytotoxic T lymphocytes (CTL) and no longer require formal antigen presentation by professional antigen presenting cells (APC). Subsequently, any cell expressing MHC class I/cognate peptide can stimulate CTL. Using TIL specific for a melanoma antigen-derived peptide, IMDQVPFSV (g209 2M), we sought to determine whether these CTL could present peptide to each other. Our findings demonstrate that peptide presentation of the g209 2M peptide epitope by TIL is comparable to conventional methods of using T2 cells as APC. We report here that CTL are capable of self-presentation of antigenic peptide to neighboring CTL resulting in IFN-gamma secretion, proliferation, and lysis of peptide-loaded CTL. These results demonstrate that human TIL possess both APC functions as well as cytotoxic functions and that this phenomenon could influence CTL activity elicited by immunotherapy. PMID- 12088414 TI - Plasmid DNA induces increased lymphocyte trafficking: a specific role for CpG motifs. AB - Bacterial DNA, primarily through immunostimulatory cytosine-guanine (CpG) motifs, induces the secretion of cytokines and activates a variety of effector cells. We investigated the possibility that CpG motifs might also modulate immunosurveillance by altering cell trafficking through a regional lymph node. Intradermal injection of plasmid DNA induced rapid and prolonged increases in the number of lymphocytes collected in efferent lymph. This effect on cell trafficking was not dependent on the expression of an encoded reporter gene but varied with plasmid construct and required a circular form. Injection of synthetic oligodeoxyribonucleotides containing CpG motifs did not alter lymphocyte trafficking but CpG-enhanced plasmid induced a dose-dependent increase in cell trafficking. Phenotypic analyses revealed that the increase in cell trafficking involved all lymphocyte subpopulations and represented a mass movement of cells. These observations reveal that bacterial DNA, through immunostimulatory CpG motifs, alters immunosurveillance by increasing cell recruitment to a regional lymph node. PMID- 12088413 TI - Stimulation of T-Cell activation by CXCL12/stromal cell derived factor-1 involves a G-protein mediated signaling pathway. AB - Recently we found that CXCL12/SDF-1 is a costimulator of peripheral CD4+ T cells. In this study, we report that CXCL12 alone induced expression of activation markers by peripheral CD4+ memory T cells and costimulated activation marker expression by anti-CD3 stimulated peripheral CD4+ naive and CD4+ memory T cells as well as by peripheral CD8+ T cells. The stimulation by CXCL12 was inhibited by Pertussis Toxin (PTX), but not by anti-CD25 mAb. CXCL12 also induced enhancement of IL-2 production and proliferation by anti-CD3 stimulated CD4+ memory T cells, but not by CD4+ naive T cells. PTX inhibited the enhancement of IL-2 production and proliferation, whereas anti-CD25 mAb inhibited proliferation, but not IL-2 production. Thus, CXCL12 upregulated T-cell activation, and a G-coupled protein mediated signaling pathway was necessary for stimulation of T cells by CXCL12. PMID- 12088415 TI - Intercellular exchange of class II major histocompatibility complex/peptide complexes is a conserved process that requires activation of T cells but is constitutive in other types of antigen presenting cell. AB - Activated T cells acquire antigen presenting cell- (APC) derived class II major histocompatibility complex glycoproteins (MHCII) but the role of TCR in this process is controversial. This study provides additional evidence that ligation of TCR initiates activation-dependent processes that independently mediate acquisition of APC-derived molecules. First, intercellular exchange of MHCII resulted in the constitutive accumulation of xenogeneic rat I-A on murine B cells, whereas naive murine T cells required activation to adsorb xenogeneic I-A. Likewise, continuous lines of B cells, basophils, and MO from various species such as rat, mouse, and human constitutively acquired xenogeneic I-A. Second, inhibitors of T-cell activation such as wortmannin, EGTA, or mAb against I-A, TCR, LFA-1, or CD4 inhibited I-A acquisition by rested T cells but not by preactivated T cells. In conclusion, exchange of MHCII is a conserved process that requires activation of T cells but is constitutive in other types of APC. PMID- 12088416 TI - Measles virus exploits dendritic cells to suppress CD4+ T-cell proliferation via expression of surface viral glycoproteins independently of T-cell trans infection. AB - Dendritic cells (DC) have been proposed to play a pivotal role in transient immune suppression induced by measles virus (MV) infection. In the present study, we show that DC-induced suppression of T-cell proliferation was not mediated by IL-10 or IFNalpha/beta, which are released following infection of DC, but required cell contacts between MV-infected DC and T cells. Human sera containing neutralizing anti-MV antibodies, as well as anti-MV hemagglutinin (HA) or fusion protein (F) mAbs, were found (i) to reverse suppression and (ii) to restore DC allostimulatory capacity. Interestingly, DC-induced T-cell suppression was associated with both phenotypic and functional DC maturation, as demonstrated by IL-12 production and chemotaxis to MIP-3beta. These data suggest that MV infection turns on the maturation program of DC allowing migration to draining lymph nodes, where potent T-cell immune suppression might be achieved via cell surface expression of HA and F glycoproteins, independently of T cell trans infection. PMID- 12088417 TI - The COX-2 inhibitor NS-398 causes T-cell developmental disruptions independent of COX-2 enzyme inhibition. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit the function of cyclooxygenases, COX-1 and COX-2, which catalyze the first step in the synthesis of inflammatory mediators (PGE2). We sought to understand the roles of cyclooxygenases and NSAIDs in T-cell development. Our data show no significant defects in T-cell development in fetal thymic organ cultures of mice disrupted in both or either COX genes or in mice disrupted in either EP-1 or EP-2 receptor genes. On the other hand, NSAIDs reproducibly caused thymocyte developmental defects. However, the specific effects of the COX-2 inhibitors were not correlated with their potency for inhibition of COX-2 activity. We focused on the NS-398 COX-2 inhibitor and showed that its effects could not be reversed by exogenous PGE2. Furthermore, NS-398 was inhibitory even when its target, COX-2, was absent. These data show that the T-cell developmental effects of NS-398 are COX-2 and PGE2 independent. PMID- 12088418 TI - Involvement of tumor necrosis factor-related apoptosis-inducing ligand in NK cell mediated and IFN-gamma-dependent suppression of subcutaneous tumor growth. AB - Natural killer (NK) cells and interferon- (IFN) gamma have been implicated in immune surveillance against tumor development. Here we show tumor necrosis factor related apoptosis-inducing ligand (TRAIL), which is a type II membrane protein belonging to the TNF family and plays a critical role in the NK cell-mediated and IFN-gamma-dependent suppression of subcutaneous growth of TRAIL-sensitive tumors. Administration of a neutralizing monoclonal antibody against TRAIL promoted outgrowth of subcutaneously inoculated TRAIL-sensitive tumors (L929, LB27.4, and Renca) but not TRAIL-resistant tumors (P815 and B16). Such a protective effect of TRAIL against TRAIL-sensitive tumors was abrogated in NK cell-depleted or IFN gamma-deficient mice. These results suggested a substantial role of TRAIL as the effector molecule that eliminates subcutaneously developing TRAIL-sensitive tumors. PMID- 12088420 TI - New isoflavone and triterpene glycosides from soybeans. AB - Chemical investigation of a soybean phytochemical concentrate resulted in the isolation and identification of two new isoflavanones, dihydrodaidzin (1) and dihydrogenistin (2), a new isoflavone, 2' ',6' '-O-diacetyloninin (3), and two new triterpenoid saponins (13 and 14). Nine known isoflavonoids (4-12) and three known saponins (15-17) were also identified. Structures of the new compounds were established by a combination of extensive NMR (DEPT, DQF-COSY, HMBC, HMQC, and ROESY) studies and chemical degradation. Cytotoxic activities (ED(50)) of various extracts and selected isoflavonoids and saponins were measured against human stomach carcinoma (Hs 740.T, Hs 756 T), breast adenocarcinoma (Hs 578 T, Hs 742.T), and prostate carcinoma (DU 145, LNCaP-FGC) cell lines. Isoflavonoids 3 and 5 were more active than 1 and 2 versus at least one of the three cell lines examined, indicating the importance of the 2,3-double bond in cytotoxicity. Saponins 13, 14, and 15 were slightly more active than saponins 16 and 17, indicating that sugar attachments at position-22 enhance cytotoxic activity. PMID- 12088419 TI - Brominated labdane-type diterpenoids from an Okinawan Laurencia sp. AB - From an unidentified species of Laurencia collected from Okinawan waters two novel brominated metabolites, 1 and 2, along with known halogenated compounds, 2,10-dibromo-3-chloro-alpha-chamigrene (3) and microcladallene A (4), were isolated and identified. The structures of these new compounds were established as ent-labdane-type bromoditerpenes, (1S,3R,5S,6S,8S,9S,10R,13R)-1-acetoxy-3 bromo-6-hydroxy-8,13-epoxy-labd-14-ene (1) and (3R,5S,6S,8S,9S,10R,13R)-3-bromo-6 hydroxy-8,13-epoxylabd-14-en-1-one (2), by interpretation of their spectroscopic data as well as by X-ray crystallographic analysis. PMID- 12088421 TI - Geographic distribution of three alkaloid chemotypes of Croton lechleri. AB - Three known alkaloids, isoboldine (2), norisoboldine (1), and magnoflorine (8), have been isolated for the first time from Croton lechleri, a source of the wound healing latex "sangre de grado". An HPLC system was developed, and a large number of latex and leaf samples of C. lechleri from 22 sites in northern Peru and Ecuador were analyzed to gain an understanding of the natural variation in alkaloid content for the species. Up to six alkaloids were found to occur in the leaves including, in addition to those listed above, thaliporphine (3), glaucine (4), and taspine (9), whereas the latex contained only 9. Taspine (9) is the component that has been previously found to be responsible for the wound healing activity of C. lechleri latex, and its mean concentration throughout the range examined was found to be 9% of the latex by dry weight. In addition, three chemotypes are defined based on the alkaloid content of the leaves, and the geographic distribution of these chemotypes is discussed along with a quantitative analysis of the alkaloid content as a function of chemotype. PMID- 12088422 TI - Antifungal stilbenoids from Stemona collinsae. AB - Fifteen new stilbenoids including 11 phenylbenzofurans, the stemofurans A-K (1 11), and four dihydrostilbenes, the stilbostemins A (15), C (17), E (19), and F (20), were isolated and identified from a methanolic extract of Stemona collinsae roots together with five known derivatives, the stilbenes pinosylvin (13) and 4' methylpinosylvin (14), the dihydrostilbenes, stilbostemins B (16) and D (18), and the dihydrophenanthrene racemosol (12) as well as (+)-sesamin, coniferyl alcohol, and stigmasterol. Bioautographic tests with Cladosporium herbarum displayed antifungal activity for stilbenoids of all four structural types. Ten derivatives were tested against five microfungi using the microdilution technique linked with digital image analysis of germ tubes. PMID- 12088423 TI - Phytotoxic compounds from Prionosciadium watsoni. AB - Bioassay-guided fractionation of a phytotoxic extract of Prionosciadium watsoni led to the isolation of three new pyranocoumarins and two pyranochromones. The new compounds were characterized as propionic acid (9R,10R)-9-acetoxy-8,8 dimethyl-9,10-dihydro-2H,8H-benzo[1,2-b:3,4-b']dipyran-2-one-10-yl ester (1), isobutyric acid (9R,10R)-9-hydroxy-8,8-dimethyl-9,10-dihydro-2H,8H-benzo[1,2 b:3,4-b']dipyran-2-one-10-yl ester (2), isobutyric acid (9R)-8,8-dimethyl-9,10 dihydro-2H,8H-benzo[1,2-b:3,4-b']dipyran-2-one-9-yl ester (10), 2-methylbut-(2Z) enoic acid (3R)-5-methoxy-3,4-dihydro-2,2,8-trimethyl-6-oxo-2H,6H-benzo[1,2-b:5,4 b']dipyran-3-yl ester (11), and isobutyric acid (3R)- 5-methoxy-3,4-dihydro-2,2,8 trimethyl-6-oxo-2H,6H-benzo[1,2-b:5,4-b']dipyran-3-yl ester (12) by spectroscopic and chemical methods. The stereochemistry at the stereogenic centers was established by applying the Mosher ester methodology. The structures of 1 and 2 were corroborated by single-crystal X-ray diffraction studies. The phytotoxic activity of the isolated compounds was assessed on Amaranthus hypochondriacus, Echinochloa crus-galli, and Lemna pausicostata. The phytotoxins also modified the electrophoretic mobility of calmodulin from both bovine-brain and spinach. PMID- 12088424 TI - Goniolactones A-F, six new styrylpyrone derivatives from the roots of Goniothalamus cheliensis. AB - Six new styrylpyrone derivatives, goniolactones A-F (1-6), have been isolated from the roots of Goniothalamus cheliensis. The structures and stereochemistry of the new compounds were elucidated by interpretation of spectroscopic data and chemical methods. The relative configuration of goniolactone A (1) was determined by X-ray crystallography analysis, and the absolute configurations of goniolactones A (1) and B (2) were established by Mosher's method. Goniolactone B (2) exhibited significant cytotoxicity against A2780, HCT-8, and KB cells with IC(50) values of 7.40, 4.43, and 7.23 microM, respectively. PMID- 12088426 TI - New nitrogenous eudesmane-type compounds isolated from the Caribbean sponge Axinyssa ambrosia. AB - Fractionation of an acetone-methanol (1:1) extract of the Caribbean marine sponge Axinyssa ambrosia yielded three new sesquiterpenes whose structures were established by spectroscopic methods as (4R*,5R*,7S*,10R*)-eudesm-11-en-4-ylamine hydrochloride (1), axinyssamine hydrochloride, (4R*,5R*,7S*,10R*)-4 isocyanatoeudesm-11-ene (3), and (4R*,5R*,7S*,10R*)-formamidoeudesm-11-ene (4). Compound 1 exhibited significant cytotoxic activity against cancer cells and was also active in a lethality test using polyps of the scleractinian coral Madracis mirabilis. PMID- 12088425 TI - Evaluation of the potential cancer chemotherapeutic efficacy of natural product isolates employing in vivo hollow fiber tests. AB - The hollow fiber test has been developed for the preliminary in vivo assessment of cancer chemotherapeutic efficacy of selected natural products. Using this model, we have established growth conditions for HL-60, HUVEC, Ishikawa, KB, KB V1, LNCaP, Lu1, MCF-7, Mel2, P-388, and SW626 cells implanted at the intraperitoneal (i.p.) and subcutaneous (s.c.) compartments of athymic mice. Five cytotoxic natural product isolates (2-6) were tested in this model, along with paclitaxel (taxol) (1). Among the compounds tested, dioscin (2) and 13-methoxy-15 oxozoapatlin (3) were found to be active, indicating their potential to function as cancer chemotherapeutic agents. On the other hand, ochraceolide A (4), alpha lapachone (5), and 2-(1-hydroxyethyl)naphtha[2,3-b]furan-4,9-quinone (6), all of which were significantly cytotoxic to cultured mammalian cells, did not mediate significant responses with the hollow fiber model. In further xenograft studies using KB cells implanted at the subcutaneous site, compound 3 mediated a statistically significant response which was consistent with the response observed at the subcutaneous compartment in the hollow fiber tests. In sum, these studies illustrate the usefulness of the hollow fiber model in natural product drug discovery programs. Preliminary indications of potential therapeutic efficacy can be provided quickly at relatively low expense. Agents capable of mediating a response at the subcutaneous site would appear to warrant greatest attention. PMID- 12088427 TI - New sesquiterpenoids from the root of Guatteria multivenia. AB - A phytochemical investigation of the CHCl(3) fraction of an ethanol extract of the root of Guatteria multivenia furnished nine compounds, of which four are sesquiterpenes (1-4) and five are alkaloids (5-9). Of the four sesquiterpenes, two are new (1, 3), named guatterin A (1) and dihydromadolin-K (3), and two are known (2, 4), identified as madolin-K (2) and madolin-W (4). The five known alkaloids were identified as liriodenine (5), lysicamine (6), lanuginosine (7), guadiscine (8), and O-methylpallidine (9). All the known compounds were isolated from this species for the first time. Structures of the new compounds were determined by extensive NMR studies, including DEPT, COSY, HMQC, HMBC, and NOESY. Compound 7 showed weak inhibitory effect against Candida albicans secreted aspartic proteases (SAP) with IC(50) of 45 microg/mL. Compound 5 was found to have antimicrobial activity against C. albicans, Cryptococcusneoformans, Staphylococcus aureus, and methicillin-resistant S. aureus (MRS) with IC(50)/MIC values of 3.5/6.25, 2.0/12.5, 2.0/3.13, and 2.0/3.13 microg/mL, respectively. PMID- 12088428 TI - Cytotoxic and antifungal triterpene glycosides from the Patagonian sea cucumber Hemoiedema spectabilis. AB - Two new sulfated triterpene glycosides, hemoiedemosides A (1) and B (2), have been isolated from the Patagonian sea cucumber Hemoiedema spectabilis. Their structures have been established by a combination of spectroscopic analysis (NMR and FABMS) and chemical transformations. Both glycosides present the same aglycon and differ in the degree of sulfation of the tetrasaccharide chain. Hemoiedemoside B (2) is a new example of a small number of trisulfated triterpene glycosides from sea cucumbers belonging to the family Cucumariidae. Glycosides 1 and 2 exhibit considerable antifungal activity against the phytopathogenic fungus Cladosporium cucumerinum, while the semisynthetic desulfated derivative 1a is less active. PMID- 12088429 TI - Structure and absolute stereochemistry of hectochlorin, a potent stimulator of actin assembly. AB - Hectochlorin (1) was isolated from marine isolates of Lyngbya majuscula collected from Hector Bay, Jamaica, and Boca del Drago Beach, Bocas del Toro, Panama. The planar structure was deduced by one- and two-dimensional NMR spectroscopy. X-ray crystallography was used to determine the absolute stereochemistry of hectochlorin as 2S,3S,14S,22S. Hectochlorin is equipotent to jasplakinolide (5) in its ability to promote actin polymerization, but unlike jasplakinolide, is unable to displace a fluorescent phalloidin analogue from polymerized actin. In addition, hectochlorin shows both a unique profile of cytotoxicity by the COMPARE algorithm and potent inhibitory activity toward the fungus Candida albicans. Structurally, hectochlorin resembles dolabellin and the recently reported lyngbyabellin class of compounds. PMID- 12088430 TI - New antimicrobial cycloartane triterpenes from Acalypha communis. AB - Three new cycloartane-type triterpenes, 16 alpha-hydroxymollic (1), 15 alpha hydroxymollic (2), and 7 beta,16 beta-dihydroxy-1,23-dideoxyjessic acids (3), were isolated from the aerial parts of Acalypha communis. The structures of the novel triterpenes were determined by spectroscopic methods as well as chemical derivatization. These compounds were tested for their antimicrobial activity against Gram-positive and -negative bacteria. Compounds 1-3 exhibited moderate antimicrobial activity (MIC 8, 32, 8 microg/mL, respectively) against vancomycin resistant enterococci. In addition, compound 1 was found to be active against methicillin-resistant staphylococci. In contrast, compounds 1-3 were poorly active against Gram-negative bacteria. Compound 3 was tested in an in vivo model; it did not provide protection to mice infected with Staphylococcus aureus. PMID- 12088431 TI - Biologically active polyketide metabolites from an undetermined fungicolous hyphomycete resembling Cladosporium. AB - Eight new polyketide-derived metabolites [cladoacetals A and B (1 and 2), 3-(2 formyl-3-hydroxyphenyl)propionic acid (3), 3-deoxyisoochracinic acid (4), isoochracinol (5), 7-hydroxy-3-(2,3-dihydroxybutyl)-1(3H)-isobenzofuranone (6), (+)-cyclosordariolone (10), and altersolanol J (11)] and six known metabolites [two isomeric 1-(1,3-dihydro-4-hydroxy-1-isobenzofuranyl)butan-2,3-diols (7a/b), 7-hydroxy-1(3H)-isobenzofuranone (8), isoochracinic acid (9), altersolanol A (12), and macrosporin (13)] have been isolated from solid-substrate fermentation cultures of an undetermined fungicolous isolate (NRRL 29097) that resembles Cladosporium sp. All structures were assigned primarily by analysis of 1D and/or 2D NMR data. Five of the compounds showed antibacterial activity. PMID- 12088432 TI - Glycolipids from sponges. 11. Isocrasserides, novel glycolipids with a five membered cyclitol widely distributed in marine sponges. AB - In addition to the previously reported crasserides (1a-m), marine sponges contain smaller amounts of the isomeric isocrasserides (3a-m). The structures of these new glycoglycerolipid analogues were determined by NMR analysis and chemical degradation. Crasserides and isocrasserides were present in all the species of marine sponges we analyzed and therefore appear to be characteristic of the phylum Porifera. PMID- 12088433 TI - Suberosols A-D, four new sesquiterpenes with beta-caryophyllene skeletons from a Taiwanese gorgonian coral Subergorgia suberosa. AB - Four new beta-caryophyllene-derived sesquiterpenes alcohols, suberosols A (1), B (2), C (3), and D (4), along with two known beta-caryophyllene-derived sesqueterpene ketones, buddledins C (5) and D (6), were isolated from a Taiwanese gorgonian coral Subergorgia suberosa. The structures of 1-4 were determined on the basis of extensive spectroscopic analyses. Cytotoxicity of these compounds toward various cancer cell lines is also described. PMID- 12088434 TI - Isolation and structure elucidation of radical scavengers from Thymus vulgaris leaves. AB - 2,2-Diphenyl-1-picrylhydrazyl radical (DPPH*) scavenging activity-guided fractionation of a leaf extract of Thymus vulgaris led to the isolation of the radical scavengers rosmarinic acid 1, eriodictyol, taxifolin, luteolin 7 glucuronide, p-cymene 2,3-diol, p-cymene 2,3-diol 6-6'-dimer, carvacrol, thymol, and a new compound, 2. The fractionation was considerably facilitated by using an on-line HPLC detector for radical scavenging activity. In this detector activity is monitored as the disappearance of the color of a postcolumn added stable radical after reacting with radical scavengers in a reaction coil. Compound 2, which consists of rosmarinic and caffeic acid moieties linked via a C-3'-C-8' ' ether bridge, was mainly elucidated by various NMR techniques and CD. Phenylpropanoid trimer 2 was a weaker and stronger radical scavenger than rosmarinic acid 1 in off-line TEAC and DPPH* assays, respectively. PMID- 12088435 TI - Gymnasterkoreaynes A-F, cytotoxic polyacetylenes from Gymnaster koraiensis. AB - Six new polyacetylenes, gymnasterkoreaynes A-F (1-6), were isolated from the roots of Gymnaster koraiensis, together with 2,9,16-heptadecatrien-4,6-diyn-8-ol (7) and 1,9,16-heptadecatriene-4,6-diyn-3,8-diol (8), by bioassay-guided fractionation using the L1210 tumor cell line as a model for cytotoxicity. The structures of compounds 1-6 were established spectroscopically, which included 2D NMR experiments. Gymnasterkoreaynes A-F (1-6) are linear diacetylenes and are structurally related to falcarinol, panaxynol, panaxydiol, and panaxytriol. Of the compounds isolated, gymnasterkoreaynes B (2), C (3), F (6), and 1,9,16 heptadecatrien-4,6-diyn-3,8-diol (8) exhibited significant cytotoxicity against L1210 tumor cells with ED(50) values of 0.12-3.3 microg/mL. PMID- 12088436 TI - Syntheses of two cytotoxic sinapyl alcohol derivatives and isolation of four new related compounds from Ligularia nelumbifolia. AB - Phytochemical reinvestigation on Ligularia nelumbifolia afforded four novel sinapyl alcohol analogues named nelumols B-E (1-4) and three known sinapyl alcohol derivatives (5-7). Their structures were elucidated by NMR techniques. Total syntheses of cytotoxic geranyloxy sinapyl alcohol (6) and geranyloxy sinapyl aldehyde (7) were carried out via two different paths. The 4-O-benzyl substituted analogues (20 and 27) as well as the 4-O-(2-methylbutenyl) derivatives (34 and 35) were also synthesized. The cytotoxicities of 6 and 7 were measured using A-549, HL-60, and KB cancer cell lines. PMID- 12088437 TI - Identification, syntheses, and characterization of the geometric isomers of 9,11 hexadecadienal from female pheromone glands of the sugar cane borer Diatraea saccharalis. AB - Chemical analysis of the pheromone glands of the sugar cane borer Diatraea saccharalis has shown the presence of the four geometric isomers of 9,11 hexadecadienal (1-4), in addition to hexadecanal and (Z)-hexadec-11-enal. We here report the syntheses and characterization of compounds 1-4. One starting material, 9-decen-1-ol, has been used to obtain all of them via divergent synthetic routes. PMID- 12088438 TI - Decipinin A and decipienolides A and B: new bioactive metabolites from the coprophilous fungus Podospora decipiens. AB - Decipinin A (1), a new compound with antifungal and antibacterial activity, has been isolated from liquid cultures of the coprophilous fungus Podospora decipiens (JS 270). Two new tetracyclic sesquiterpene lactones, decipienolides A (2) and B (3), were also obtained from this isolate as an inseparable mixture of epimers that showed antibacterial activity. The structures of 1-3 were elucidated by analysis of 1D and 2D NMR data, aided by chemical shift comparisons to related compounds. PMID- 12088439 TI - Spiroidesin, a novel lipopeptide from the cyanobacterium Anabaena spiroides that inhibits cell growth of the cyanobacterium Microcystis aeruginosa. AB - Spiroidesin (1), a novel D-amino acid-containing linear lipopeptide, was isolated from waterblooms of the cyanobacterium Anabaena spiroides. The structure was identified by 2D NMR and chemical degradation analyses. Spiroidesin inhibited cell growth of the toxic cyanobacterium Microcystis aeruginosa (IC(50), 1.6 x 10( 6) M). PMID- 12088440 TI - Callyspongynic acid, a polyacetylenic acid which inhibits alpha-glucosidase, from the marine sponge Callyspongia truncata. AB - A new polyacetylenic acid, callyspongynic acid (1), was isolated as an alpha glucosidase inhibitor from the marine sponge Callyspongia truncata. Its structure was determined by spectroscopic and chemical methods. PMID- 12088441 TI - Lyngbouilloside, a novel glycosidic macrolide from the marine cyanobacterium Lyngbya bouillonii. AB - A glycosidic macrolide, lyngbouilloside (1), was characterized from the marine cyanobacterium Lyngbya bouillonii collected from Papua New Guinea. Its structure was elucidated by spectroscopic analysis and chemical derivatization. Relative stereochemistry was deduced from homonuclear and heteronuclear coupling constants as well as NOE information. Lyngbouilloside was modestly cytotoxic to neuroblastoma cells (IC(50) value of 17 microM). PMID- 12088442 TI - New tropane alkaloids from Erythroxylum moonii. AB - Four new tropane alkaloids were isolated from the leaves of Erythroxylum moonii and identified as 3 alpha-isobutyryloxy-7 beta-hydroxynortropane (1), 3 alpha hydroxy-7 beta-phenylacetoxynortropane (2), 3 alpha-cis-cinnamoyloxytropane (3), and 3 alpha-hydroxy-6 beta-(3'-hydroxy-2'-methyl-3'-phenylpropionyloxy)-7 beta hydroxytropane (4). Other alkaloids isolated for the first time from E. moonii were 3 alpha-benzoyloxytropane, 3 alpha-phenylacetoxytropane, 3 alpha-trans cinnamoyloxytropane, and 3 alpha-phenylacetoxy-6 beta,7 beta-dihydroxynortropane. The structures of compounds 1-4 were elucidated by spectroscopic methods. PMID- 12088443 TI - Novel diterpene lactones from Suregada multiflora. AB - Two new diterpene lactones, suregadolides A (1) and B (2), were isolated from a dichloromethane extract of Suregada multiflora bark. These compounds possess a novel skeleton, which contains a cyclopropane ring bridging C-3 and C-4 of the abietane skeleton. The structures were established on the basis of one- and two dimensional NMR and other spectroscopic studies. Compound 1 showed moderate inhibitory activity in a mutant yeast strain bioassay. PMID- 12088444 TI - Ecteinascidins 770 and 786 from the Thai tunicate Ecteinascidia thurstoni. AB - Ecteinascidins 770 (1b) and 786 (3b) were isolated from the pretreated Thai tunicate Ecteinascidia thurstoni with potassium cyanide in buffer solution (pH 7). These structures were fully elucidated by extensive 2D NMR analysis. PMID- 12088445 TI - Convolutindole A and convolutamine H, new nematocidal brominated alkaloids from the marine bryozoan Amathia convoluta. AB - Nematocidal activity of an extract of the marine bryozoan Amathia convoluta, collected from Tasmania's east coast, was ascribed to two novel tribrominated alkaloids: convolutamine H (2) and convolutindole A (5), an indole possessing the unusual N-methoxy moiety. The structures were established by spectroscopic techniques. PMID- 12088447 TI - Cytotoxic sphingosine 4-sulfates from the sponge Spirastrella abata. AB - Two new sphingosine 4-sulfates (1 and 2) have been isolated from the sponge Spirastrella abata as cytotoxic constituents. The gross structures of 1 and 2 were determined by spectroscopic analysis, and their stereochemistry was established by chemical conversion. The compounds exhibited significant cytotoxicity against a small panel of five human tumor cell lines. PMID- 12088446 TI - Isolation of a new mycalolide from the marine sponge Mycale izuensis. AB - Bioassay-directed fractionation of the lipophilic extract of the marine sponge Mycale izuensis led to the isolation of cytotoxic mycalolides including a new compound, 30,32-dihydroxymycalolide A (1). Its structure including absolute stereochemistry was deduced by spectroscopic and chemical methods. Compound 1 was cytotoxic against HeLa cells with an IC(50) value of 2.6 ng/mL. PMID- 12088448 TI - Oxachamigrenes, new halogenated sesquiterpenes from Laurencia obtusa. AB - Two new sesquiterpenes belonging to a novel oxacyclic structural type of chamigrene skeleton, oxachamigrene (1) and 5-acetoxyoxachamigrene (2), have been isolated from the red alga Laurencia obtusa. The structures of the compounds were determined on the basis of spectroscopic evidence. A biogenetic route for these metabolites has been proposed. PMID- 12088449 TI - Clinical laboratory assays for HER-2/neu amplification and overexpression: quality assurance, standardization, and proficiency testing. AB - OBJECTIVE: To present and contrast the results of immunohistochemistry and fluorescence in situ hybridization (FISH) proficiency testing surveys for HER 2/neu, as conducted by the Cell Markers and Cytogenetics Committees of the College of American Pathologists. DESIGN: During the past 2 years, unstained sections from invasive breast carcinomas have been used for both immunohistochemistry and interphase FISH proficiency surveys. In most instances, the same cases were used for both the Cell Markers and Cytogenetics surveys. Additional data regarding interpretative variability for immunohistochemistry surveys have also been captured. RESULTS: The number of laboratories performing FISH for HER-2/neu testing doubled during the 2-year period. The results of FISH testing have been remarkably concordant for laboratories submitting results. In contrast, the results of immunohistochemistry testing continue to highlight substantial variability among laboratories evaluating the same cases. The data show that this variability is at least in part due to variability in interpretation among observers, as well as inherent differences between immunohistochemistry and FISH techniques. CONCLUSIONS: College of American Pathologists proficiency survey programs provide useful information about clinical testing for HER-2/neu amplification/overexpression. PMID- 12088450 TI - Continuous wristband monitoring over 2 years decreases identification errors: a College of American Pathologists Q-Tracks Study. AB - CONTEXT: Identification of patients is one of the first steps in ensuring the accuracy of laboratory results. In the United States, hospitalized patients wear wristbands to aid in their identification, but wristbands errors are frequently found. OBJECTIVE: To investigate if continuous monitoring of wristband errors by participants of the College of American Pathologists (CAP) Q-Tracks program results in lower wristband error rates. SETTING: A total of 217 institutions voluntarily participating in the CAP Q-Tracks interlaboratory quality improvement program in 1999 and 2000. DESIGN: Participants completed a demographic form, answered a questionnaire, collected wristband data, and at the end of the year, best and most improved performers answered another questionnaire seeking suggestions for improvement. Each institution's phlebotomists inspected wristbands for errors before performing phlebotomy and recorded the number of patients with wristband errors. On a monthly basis, participants submitted data to the CAP for data processing, and at the end of each quarter, participants received summarized comparisons. At the end of each year, participants also received a critique of the results along with suggestions for improvement. MAIN OUTCOME MEASURES: The percentage of wristband errors by quarter, types of wristband errors, and suggestions for improvement. RESULTS: During 2 years, 1 757 730 wristbands were examined, and 45 197 wristband errors were found. The participants' mean wristband error rate for the first quarter in 1999 was 7.40%; by the eighth quarter, the mean wristband error rate had fallen to 3.05% (P <.001). Continuous improvement occurred in each quarter for participants in the 1999 and 2000 program and in 7 of 8 quarters for those who participated in both 1999 and 2000. Missing wristbands accounted for 71.6% of wristband errors, and best performers usually had wristband error rates under 1.0%. The suggestion for improvement provided by the largest number of best and most improved performers was that phlebotomists should refuse to perform phlebotomy on a patient when a wristband error is detected. CONCLUSIONS: The wristband error rate decreased markedly when this rate was monitored continuously using the CAP Q-Tracks program. The Q-Tracks program provides a useful tool for improving the quality of services in anatomic pathology and laboratory medicine. PMID- 12088451 TI - Tyrosinase expression in malignant melanoma, desmoplastic melanoma, and peripheral nerve tumors. AB - CONTEXT: Pathologists may encounter problems in the differential diagnosis of malignant melanoma, spindle and epithelioid neoplasms of peripheral nerves, and fibrohistiocytic tumors. Tyrosinase has been demonstrated to be a sensitive marker for melanoma. OBJECTIVE: To determine the specificity of tyrosinase expression in the differential diagnosis of melanoma, desmoplastic melanoma, and peripheral nerve sheath tumors. DESIGN: Immunoreactivity for tyrosinase, HMB-45 (anti-gp100 protein), S100 protein, CD34, and vimentin was studied in 70 tumors, including 15 melanomas (5 desmoplastic, 4 amelanotic, 6 melanotic), 13 malignant peripheral nerve sheath tumors; 10 schwannomas (1 pigmented), 12 neurofibromas (4 pigmented), and 20 fibrohistiocytic tumors (10 dermatofibrosarcoma protuberans and 10 dermatofibromas). Microwave-based antigen retrieval was performed in 10mM citrate buffer, pH 6.0, for 20 minutes at 121 degrees C. RESULTS: All melanomas demonstrated positive immunostaining for tyrosinase, HMB-45, and S100 protein. Immunoreactivity for HMB-45 was generally stronger than that for tyrosinase in amelanotic lesions and significantly stronger in 1 of the desmoplastic lesions. The 4 pigmented neurofibromas were focally positive for tyrosinase, but did not stain for HMB-45. The pigmented schwannoma was focally positive for both tyrosinase and HMB-45. The malignant peripheral nerve sheath tumors, dermatofibrosarcoma protuberans, and dermatofibromas were nonreactive for tyrosinase and HMB-45. CONCLUSIONS: Our results support the sensitivity of tyrosinase expression and demonstrate the relative specificity of tyrosinase as a marker for melanocytic lesions, including desmoplastic melanoma, although pigmented peripheral nerve tumors may demonstrate focal positive staining. Immunoreactivity for tyrosinase and HMB-45 may have been enhanced by the microwave-based antigen-retrieval technique used in this study. PMID- 12088452 TI - CD34/QBEND10 immunostaining in the bone marrow trephine biopsy: a study of CD34 positive mononuclear cells and megakaryocytes. AB - CONTEXT: The immunohistochemical detection of CD34 protein using QBEND10 antibody in bone marrow trephine biopsies was shown recently to be a precise method for quantitation of blasts and a possibly useful approach in diagnosis and classification of myelodysplastic syndrome. OBJECTIVES: To evaluate CD34+ cells in bone marrow biopsies with various diagnoses and to assess how counts obtained using this method correlate with blast counts obtained by traditional morphologic evaluation of bone marrow smears. DESIGN: Bone marrow trephine biopsies from 108 adult patients were evaluated by immunohistochemistry using anti-CD34 antibody (QBEND10). CD34+ mononuclear cells were counted and compared with the blast counts in the bone marrow aspirate smears or imprints. CD34+ mononuclear cell clusters and CD34+ megakaryocytes were also recorded. The type of positivity (membranous vs cytoplasmic) and the percentage of CD34+ megakaryocytes were evaluated because the presence of CD34+ megakaryocytes was recently suggested to be present in myelodysplastic syndrome, but not in myeloproliferative disease or nonneoplastic bone marrow. RESULTS: Six of 24 biopsies with partial involvement by non-Hodgkin lymphoma and 5 of 60 biopsies with reactive changes had 5% to 10% CD34+ mononuclear cells and were associated with lymphocytosis and increased hematogones. The CD34+ mononuclear cell clusters were found only in myelodysplastic syndrome and myeloproliferative disease. The CD34+ megakaryocytes were present in all diagnostic groups. CONCLUSION: The number of CD34+ mononuclear cells was often slightly higher than the number of myeloid blasts in the bone marrow smears, probably due to increased hematogones. The presence and the number of CD34+ megakaryocytes do not appear to have diagnostic value, but this finding should be further investigated in relation to clinical parameters. PMID- 12088453 TI - Immunohistochemical study of myofibroblasts in normal colonic mucosa, hyperplastic polyps, and adenomatous colorectal polyps. AB - CONTEXT: Myofibroblasts are distinct cells with characteristics of both smooth muscle cells and fibroblasts. Through their ability to secrete cytokines, chemokines, prostaglandins, growth factors, and matrix components, they are thought to play critical roles in inflammation, growth, repair, and neoplasia. OBJECTIVE: The goal of this study was to identify the distinct cell populations of the lamina propria of normal colon and colorectal polyps. DESIGN: We studied the expression of alpha-smooth muscle actin (alphaSMA), smooth muscle myosin (SMM), desmin, vimentin, and c-kit by intestinal mesenchymal (stromal) cells in the normal colonic mucosa (n = 5), as well as in hyperplastic polyps (n = 5), sporadic colorectal adenomas (n = 47), and adenomas from patients with familial polyposis (n = 36). RESULTS: In the normal colonic mucosa, the pericryptal stromal cells were alphaSMA+, SMM+, desmin-, and vimentin+, defining them as myofibroblasts. In contrast, cells of the muscularis mucosae were alphaSMA+, SMM+, desmin+, and vimentin-, defining them as smooth muscle cells. alpha-Smooth muscle actin also highlighted direct connections between the muscularis mucosae and the pericryptal myofibroblasts, and vimentin immunostaining showed a network of connections between the alphaSMA+ pericryptal myofibroblasts and the alphaSMA- fibroblasts in the interstitium. In all hyperplastic polyps and adenomatous polyps, the interstitial stromal cells (fibroblasts) now also express alphaSMA and form a syncytium of alphaSMA+ networklike connections throughout the lamina propria. Stromal cells of sporadic adenomas demonstrated the same immunohistochemical staining characteristics displayed by adenomas from patients with familial polyposis and by hyperplastic polyps. Conclusions.-These findings indicate that in normal colon, alphaSMA- fibroblasts are the predominant cell type in the lamina propria. However, the pericryptal (subepithelial) stromal cells are a distinct cell type (alphaSMA+ myofibroblast) that is immunophenotypically different from muscularis mucosae smooth muscle cells and are connected to the interstitial, nonpericryptal fibroblasts with which they exist as a network throughout the lamina propria of the normal colon. Furthermore, in both hyperplastic and neoplastic polyps, there are changes in nonpericryptal fibroblasts from vimentin+, alphaSMA-, and SMM- to vimentin+, alphaSMA+, and SMM+; thus, the interstitial fibroblasts are replaced by myofibroblasts. The factors that cause these changes and the origin of the myofibroblasts need to be determined to clarify the biology of colorectal tumorigenesis. PMID- 12088454 TI - Epithelial neoplasms of the appendix and colorectum: an analysis of cell proliferation, apoptosis, and expression of p53, CD44, bcl-2. AB - CONTEXT: Carcinomas of the appendix are usually well-differentiated mucinous adenocarcinomas that tend to produce pseudomyxoma peritonei and do not show metastatic spread until late in the disease process. In contrast, adenocarcinomas of the colon and rectum rarely result in pseudomyxoma peritonei and frequently metastasize, even if mucinous and well differentiated. These differences in behavior may be reflected by differences at the molecular level. OBJECTIVES: To examine adenocarcinomas and their precursor lesions (adenomas) of the appendix and colorectum and to determine whether differences exist in the numbers of proliferating and apoptotic cells or in expression of p53, bcl-2, and the standard form of CD44 (CD44s). DESIGN: Retrospective analysis of surgical specimens. SETTING: Multicenter study. PATIENTS: Individuals treated surgically for tumors of the appendix or colorectum. INTERVENTIONS: Sections were cut from formalin-fixed surgical specimens and immunohistochemical tests were performed for Ki-67 (as a marker of proliferating cells), M30 (as a marker of apoptotic cells), p53, CD44s, and bcl-2. MAIN OUTCOME MEASURES: Expression of Ki-67, M30, p53, CD44s, and bcl-2 in tumor cells. RESULTS: The appendiceal adenomas showed significantly lower Ki-67 counts, p53 expression, and bcl-2 expression. When compared with adenocarcinomas of the colorectum in general (mucinous and nonmucinous), the appendiceal adenocarcinomas showed significantly lower Ki-67 counts, M30 counts, and CD44s expression. However, when the analysis was confined to well-differentiated mucinous adenocarcinomas, only the M30 count was significantly different. CONCLUSIONS: The lower proliferative and apoptotic activity of appendiceal carcinomas and the lower CD44s expression are in keeping with their more indolent behavior compared with adenocarcinomas of the colorectum. However, when only the subset of well-differentiated mucinous adenocarcinomas was compared, only the apoptotic activity was different, suggesting that the other differences were related to the morphologic structure of the lesions. PMID- 12088455 TI - C1 inhibitor infusion modifies platelet activity in hereditary angioedema patients. AB - CONTEXT: C1 inhibitor (C1-INH) is an alpha2-globulin that blocks esterolytic activity of the first component of the classic complement cascade. The alpha granules of normal human platelets also contain C1-INH, which is expressed on the platelet surface during platelet secretion in healthy patients, but it is clearly reduced in patients with hereditary angioedema (HAE). OBJECTIVE: To evaluate the effects of in vivo C1-INH concentrate infusion on platelet responsiveness and coagulation system activity in patients with HAE. DESIGN: Assessment of the platelet activity and plasma levels of C1-INH, activated factor XII (XIIa), and prothrombin fragment F1.2 (F1.2) before and after infusion of 15 U/kg of C1-INH concentrate. PATIENTS: In 6 patients (4 men and 2 women), HAE was diagnosed according to the accepted clinical and laboratory criteria. MEASUREMENTS: Platelet aggregation (final concentrations: adenosine diphosphate, 0.5, 1.25, and 2.5 microM; collagen, 5 microg/mL), C1-INH antigen (radial immunodiffusion), C1 INH activity (chromogenic substrates), and XIIa and F1.2 (enzyme-linked immunosorbent assay). RESULTS: After C1-INH infusion, we observed a prompt increase of C1-INH level and a slow return toward its plasma preinfusion values within 4 to 7 days, a significant decrease of both adenosine diphosphate- and collagen-induced platelet aggregation versus preinfusion values (maximum after 1 2 days; P <.001), and a rapid decrease of high basal values of XIIa and F1.2 in 30 and 120 minutes, respectively. CONCLUSIONS: These data show a role of C1-INH in the control of platelet activity and that its deficiency increases platelet aggregability and plasma levels of XIIa and F1.2 in patients with HAE. PMID- 12088456 TI - Intraoperative evaluation of lumpectomy margins by imprint cytology with histologic correlation: a community hospital experience. AB - BACKGROUND: Several well-controlled studies have demonstrated significantly increased local recurrence rates in patients with low-stage breast carcinoma treated with breast conservation therapy in whom focally positive margins were not reexcised. Imprint cytology is a rapid technique for evaluating surgical margins intraoperatively, thus allowing reexcisions to be performed during the initial surgery. The large majority of studies on the use of intraoperative imprint cytologic examination of breast conservation therapy margins have been performed at university-based academic centers. OBJECTIVE: To evaluate the utility of intraoperative imprint cytologic evaluation of breast conservation therapy margins in a community hospital setting. METHODS: We retrospectively reviewed the intraoperative imprint cytology margins of 141 lumpectomy specimens that had been obtained from 137 patients between May 1997 and May 2001. RESULTS: We evaluated 758 separate margins. On a patient basis, the sensitivity was 80%, the specificity was 85%, the positive predictive value was 40%, the negative predictive value was 97%, and the overall accuracy was 85%. There were no cytologically unsatisfactory margins. CONCLUSION: Imprint cytology is an accurate, simple, rapid, and cost-effective method for determining the margin status of breast conservation therapy specimens intraoperatively in the community hospital setting. This method allows a survey of the entire surface area of the lumpectomy specimen, which is not practical using frozen section evaluation. PMID- 12088457 TI - Undifferentiated sarcoma of the parotid gland with osseous metaplasia. AB - Malignant spindle cell tumors of the parotid gland are a diagnostic challenge. We present an unusual case of such a tumor that occurred in the right parotid gland of a 53-year-old man. The clinical and histologic assessments were consistent with a primary sarcoma of the parotid gland. The tumor was composed of sheets of pleomorphic, spindle-shaped cells with an area of bone formation. By immunohistochemistry, the tumor cells were positive for vimentin and negative for epithelial markers. Electron microscopy revealed mesenchymal cells containing moderate amounts of rough endoplasmic reticulum. The major differential diagnostic considerations were spindle cell carcinoma, carcinosarcoma, and primary undifferentiated sarcoma with osseous metaplasia. The lack of epithelial features and the benign appearance of the bone formation led to a diagnosis of undifferentiated sarcoma of the parotid gland. PMID- 12088458 TI - A carcinosarcoma/sarcomatoid carcinoma arising in a urinary bladder diverticulum. AB - We describe an invasive polypoid carcinosarcoma/sarcomatoid carcinoma arising within a urinary bladder diverticulum in a 65-year-old patient with synchronous, moderately differentiated prostatic adenocarcinoma. Histologically, the diverticular tumor exhibits an admixture of different morphologic components, including invasive high-grade urothelial carcinoma, malignant glandular structures in a cellular background of malignant spindle cells, and areas formed exclusively by spindle and pleomorphic cells. There was full-thickness involvement of the diverticulum with extension of the tumor into the perivesical fat and ipsilateral seminal vesicle. In view of the early invasive behavior of carcinosarcoma/sarcomatoid carcinoma combined with the paucity of the muscular layer in the diverticulum wall, a graver prognosis was expected for this aggressive tumor that occurred in this unusual site. PMID- 12088459 TI - Metastatic renal cell carcinoma presenting as a hemorrhoid. AB - Metastatic, noncolorectal carcinoma involving the anal canal is exceptionally rare, with only 3 cases being described in the medical literature. We report the case of a 53-year-old man with an anal mass clinically presenting as a large, thrombosed, internal hemorrhoid. The patient had a history of nephrectomy for renal cell carcinoma 9 years previously. The resected anal lesion was histologically identical to the primary tumor in the kidney, showing features of renal cell carcinoma of the clear cell type. To the best of our knowledge, this is only the fourth reported case of metastatic, noncolorectal carcinoma involving the anal canal and is the first report of a renal cell carcinoma metastasis to this site. PMID- 12088460 TI - Microcystic transitional cell carcinoma: a report of 2 cases arising in the renal pelvis. AB - Microcystic transitional cell carcinoma is a rare variant of urothelial carcinoma; to date, it has been described only in the urinary bladder. We report 2 cases of microcystic transitional cell carcinoma arising in the renal pelvis. The first case occurred in a 73-year-old man with a history of superficially invasive transitional cell carcinoma who presented with macroscopic hematuria and anemia. The second case occurred in a 62-year-old woman who had no relevant medical history and presented with hematuria. Computed tomographic scan revealed a tumor of the renal pelvis. In both cases, microscopic examination showed invasive transitional cell carcinoma with prominent cystic features. The cysts were irregular in size and were deeply infiltrative. The cysts were lined by single or multiple layers of cuboidal or flattened cells with minimal cytological atypia. The first patient died of his disease 18 months after presentation. The second patient remained well at her 6-month follow-up examination. Microcystic transitional cell carcinoma is an unusual, deceptively bland variant of urothelial carcinoma, which can mimic benign lesions. PMID- 12088461 TI - Who we are versus what we do. PMID- 12088462 TI - The experience of providing physical therapy in a changing health care environment. AB - BACKGROUND AND PURPOSE: The changes in the health care environment during the last decade have had an impact on the roles and responsibilities of all health care professionals. The purpose of this phenomenological study was to describe the experience of staff physical therapists during a time of systemic change within a large urban academic medical center. SUBJECTS AND METHODS: Participants were 5 physical therapists working in various clinical settings within the medical center. The participants were interviewed and asked the question "Over the past 4 years, there have been major changes in your work environment. What has it been like for you working as a clinician during this time of change?" Interviews were recorded, transcribed, and analyzed to find thematic patterns of responses. RESULTS: Four common themes emerged in which participants described experiencing loss of control, stress, discontent, and disheartenment. A fifth theme showed that despite these negative feelings, participants were able to "find the silver lining" in their daily work lives (ie, they were able to find positive aspects of their professional lives despite the perceived unpleasant changes with which they had to cope). DISCUSSION AND CONCLUSION: This study provides insight into the experiences of a group of staff physical therapists during a time of systemic change in their work environment. Although the themes reflect largely unsettling and negative experiences, there seems to be an underlying ability to find affirmative aspects of work. PMID- 12088463 TI - Effect of passive range of motion exercises on lower-extremity goniometric measurements of adults with cerebral palsy: a single-subject design. AB - BACKGROUND AND PURPOSE: People with spastic cerebral palsy often receive passive stretching that is intended to maintain or increase joint passive range of motion (PROM) even though the effectiveness of these exercises has not been definitively demonstrated. The purpose of this study was to determine the effect of PROM exercises on 6 adults with spastic quadriplegia and contractures. PARTICIPANTS: Four men and 2 women (X =31 years of age, range=20-44 years) who lived in an institution for people with mental retardation participated in the study. METHODS: The authors used 2 multiple baseline designs. Three participants (group 1) received lower-extremity PROM exercises during phase A; PROM exercises were discontinued during phase B. Three participants (group 2) did not receive PROM exercises during phase A; PROM exercises were initiated during phase B. Data were analyzed using visual analysis and the C statistic. RESULTS: Results varied with the method of analysis; however, phase A and phase B measurements, overall, did not differ for either group. DISCUSSION AND CONCLUSION: This study demonstrated use of a single-subject design to measure the effect of PROM exercises on adults with cerebral palsy. The authors concluded that the PROM exercise protocol did not have an effect on the lower-extremity goniometric measurements of the participants. PMID- 12088465 TI - Scale to measure patient satisfaction with physical therapy. AB - BACKGROUND AND PURPOSE: Patient satisfaction can be one indicator of quality of care. In this study, a patient satisfaction questionnaire for physical therapy was developed. SUBJECTS: The subjects were a consecutive sample of 1,024 patients who received physical therapy between January and March 1999 at a teaching hospital in Geneva, Switzerland. METHODS: A cross-sectional mail survey was conducted in which a structured questionnaire measuring patient satisfaction with various aspects of physical therapy followed by open-ended questions was sent to the subjects. RESULTS: Overall, 528 of 1,024 patients (52%) responded (patient demographics for 501 respondents who provided demographic data: mean years of age=58.6, SD=18.9, range=15-95; 258 men, 243 women). Factor analysis was used to identify main domains of satisfaction, and a scale was constructed to measure satisfaction with each dimension: treatment subscale (5 items), admission subscale (3 items), logistics subscale (4 items), and a global assessment subscale (2 items). All subscales had good acceptability and small floor and ceiling effects. Internal consistency coefficients varied between.77 and.90, indicating good reliability for all subscales. Scale validity was supported by a logical grouping of items into subscales, according to their content, and by correlations of satisfaction scores with the patient's intention to recommend the facility and with the number of positive and negative comments to open-ended questions. Younger patients were less satisfied than older patients for 2 of the subscales (admission and logistics). DISCUSSION AND CONCLUSION: The 14-item instrument is a promising tool for the evaluation of patient satisfaction with physical therapy in both inpatients and outpatients. PMID- 12088464 TI - Environmental demands associated with community mobility in older adults with and without mobility disabilities. AB - BACKGROUND AND PURPOSE: In this study, the influence of 8 dimensions of the physical environment on mobility in older adults with and without mobility disability was measured. This was done in order to identify environmental factors that contribute to mobility disability. SUBJECTS: Subjects were 36 older adults ((> or = 70 years of age) who were recruited from 2 geographic sites (Seattle, Wash, and Waterloo, Ontario, Canada) and were grouped according to level of mobility function (physically able [ability to walk 1/2 mile (0.8 km) or climb stairs without assistance], physically disabled). METHODS: Subjects were observed and videotaped during 3 trips into the community (trip to grocery store, physician visit, recreational trip). Frequency of encounters with environmental features within each of the 8 dimensions was recorded. Differences in baseline characteristics and environmental encounters were analyzed using an analysis of variance or the Fisher exact test, as appropriate. RESULTS: Mobility disability among older adults was not associated with a uniform decrease in encounters with environmental challenges across all dimensions. Environmental dimensions that differed between subjects who were physically able and those with physical disability included temporal factors, physical load, terrain, and postural transition. Dimensions that were not different included distance, density, ambient conditions (eg, light levels and weather conditions), and attentional demands. DISCUSSION AND CONCLUSION: Understanding the relationship of the environment to mobility is crucial to both prevention and rehabilitation of mobility disability in older adults. Among older adults, certain dimensions of the environment may disable community mobility more than others. PMID- 12088466 TI - A retrospective analysis of ethics knowledge in physical therapy (1970-2000). AB - BACKGROUND AND PURPOSE: Purtilo, Guccione, and others have noted that increased clinical autonomy for physical therapists presents more complex ethical dilemmas. The body of literature examining physical therapy ethics, however, is relatively small and has not been analyzed. The primary purposes of this research were: (1) to use multiple perspectives to describe and analyze literature examining ethics in physical therapy from 1970 to 2000, (2) to develop a model to describe the evolution of knowledge of ethics in physical therapy during this period, and (3) to compare the proposed model with the evolutionary models proposed by Purtilo in physical therapy and by Pellegrino in bioethics. SAMPLE: The sample consisted of peer-reviewed journal articles cited in the MEDLINE or Cumulative Index to Nursing and Allied Health Literature (CINAHL) databases between 1970 and 2000 or referenced in Ethics in Physical Therapy. METHODS: A two-phase mixed quantitative and qualitative method was used to analyze publications. In the quantitative phase, the author sorted publications into a priori categories, including approach to ethics, author, decade, country of publication, role of the physical therapist, and component of morality. During the qualitative phase of the research, the author analyzed and sorted the publications to identify common themes, patterns, similarities, and evolutionary trends. These findings were compared with the evolutionary models of Pellegrino and Purtilo. RESULTS: The 90 publications meeting inclusion criteria were predominantly philosophical, using the "principles" perspective; focused on the patient/client management role of the physical therapist; and addressed the moral judgment component of moral behavior. As predicted by Purtilo's model, the focus of identity evolved from self-identity to patient-focused identity, with increasing representation of societal identity. Recurrent themes included the need to further identify and clarify physical therapists' ethical dilemmas, the interrelationship between clinical and ethical decision making, and the changing relationship with patients. DISCUSSION AND CONCLUSION: Although knowledge of ethics grew steadily between 1970 and 2000, this retrospective analysis identified gaps in our current knowledge. Further research is needed to address the unique ethical problems commonly encountered in all 5 roles of the physical therapist; patient perspectives on ethical issues in physical therapy; variety in ethical approaches; factors affecting moral judgment, sensitivity, motivation, and courage; and cultural dimensions of ethical practice in physical therapy. PMID- 12088467 TI - Improved intralimb coordination in people with incomplete spinal cord injury following training with body weight support and electrical stimulation. AB - BACKGROUND AND PURPOSE: Limb coordination is an element of motor control that is frequently disrupted following spinal cord injury (SCI). The authors assessed intralimb coordination in subjects with SCI following a 12-week program combining body weight support, electrical stimulation, and treadmill training. SUBJECTS: Fourteen subjects with long-standing (mean time post-SCI=70 months, range=12-171 months), incomplete SCI participated. Three subjects without SCI provided data for comparison. METHODS: A vector-based technique was used to assign values to the frame-by-frame changes in hip/knee angle, and vector analysis techniques were used to assess how closely the hip/knee angles of each step cycle resembled those of every other step cycle. Overground and treadmill walking speeds also were measured. RESULTS: Following training, 9 of the 14 subjects with SCI demonstrated greater intercycle agreement. Mean overground and treadmill walking speeds improved (84% and 158%, respectively). DISCUSSION AND CONCLUSION: The intervention used in this study is based on our current understanding of the role of afferent input in the production of walking. Although the study sample was small and there was no control group, results suggest that training may improve intralimb coordination in people with SCI. PMID- 12088468 TI - Managing ovarian cancer. PMID- 12088469 TI - Can we improve pain management in nursing homes? PMID- 12088470 TI - Endometriosis. PMID- 12088471 TI - A medical school for the Australian National University. PMID- 12088472 TI - Ovarian cancer: patterns of care in Victoria during 1993-1995. AB - OBJECTIVE: To describe the management of and outcomes in patients with newly diagnosed ovarian cancer during 1993, 1994 and 1995 in Victoria. DESIGN AND SETTING: Retrospective cohort study conducted by surveying doctors involved in managing incident ovarian cancer cases identified from the population-based Victorian Cancer Registry. The survey was conducted in 1997 and the cohort was followed up until the end of 1999 to obtain at least four years of follow-up data on all patients. PATIENTS: All women with invasive epithelial ovarian cancer diagnosed during 1993, 1994 and 1995. MAIN OUTCOME MEASURES: Reported management in terms of staging, treatment and survival. RESULTS: Management details were obtained for 84.5% (562/665) of eligible patients. Median age at diagnosis was 66 years (range, 22-98 years). Surgery was the primary therapy in 77.2% of women (434/562). Only one in three women had adequate surgery, which was less likely to be performed by general gynaecologists and general surgeons than gynaecological oncologists (21.3% [35/164] v 13.3% [8/60] v 52% [105/202]). After surgery 78.6% of women (341/434) received chemotherapy, usually with platinum-based regimens. The overall five-year relative survival was 46% for women treated surgically; poor survival was related to increasing age, later tumour stage, presence of ascites, residual disease > 2 cm and poorer histological differentiation of the tumour. CONCLUSIONS: For optimal care a preoperative carcinoma antigen (CA)-125 assay, chest x-ray and pelvic ultrasound should be performed, and early referral to a multi-disciplinary unit for definitive surgery is advised. Every effort should be made to adequately stage or debulk the tumour. Women with high-risk early-stage and advanced disease should be considered for platinum-based chemotherapy. PMID- 12088473 TI - Prevalence of pain among nursing home residents in rural New South Wales. AB - OBJECTIVE: To measure the prevalence of pain among residents of rural and regional nursing homes in northern New South Wales and to describe the procedures used for pain management. DESIGN: Cross-sectional survey using interviews and audit of medical records. SETTING AND PARTICIPANTS: 917 nursing home residents in 15 nursing homes within a northern NSW area health service in 1998-1999. MAIN OUTCOME MEASURES: Number of residents experiencing pain at the time of interview; sites of pain and magnitude of pain problem; diagnoses relevant to pain; analgesic prescribing patterns; non-pharmacological treatments for pain; and the extent of pain documentation in nursing records. RESULTS: The prevalence of pain present at interview was 27.8% (95% CI, 21.8%-33.8%). Women reported pain more often than men (31% v 21%; chi(2)(2) = 5.38; P = 0.02), but pain was not significantly associated with age, length of stay, or diagnoses of arthritis or dementia. Common sites for pain were the limbs, joints and back; 22% of residents reporting pain had no record of analgesic medication, and 16% had had no form of pain treatment ordered. Agreement between the nursing record and the residents' pain symptoms was borderline poor/fair (kappa, 0.24). CONCLUSIONS: The prevalence of pain is high among nursing home residents in rural NSW who are able to communicate their pain. Descriptive data suggest that pain management activities could be substantially improved. PMID- 12088474 TI - Use of medication by young people with attention-deficit/hyperactivity disorder. AB - OBJECTIVES: To examine the prevalence of psychotropic medication use by children with attention-deficit/hyperactivity disorder (ADHD) and children without ADHD. To identify factors associated with stimulant use by children in the community. DESIGN: A representative, multistage probability sample of Australian households was conducted in 1998. Parents completed questionnaires assessing children's mental health problems and health-related quality of life. They also completed a structured interview to identify children's psychiatric disorders and their use of medications during the previous six months. PARTICIPANTS: Parent or main caregiver of 3597 children aged 6-17 years. MAIN OUTCOME MEASURES: Rates of use of stimulants (dexamphetamine and methylphenidate), antidepressants and clonidine by children. RESULTS: Overall, 1.8% of children (95% CI, 1.5%-2.3%) were receiving stimulant medication. Of those with ADHD, 12.6% (95% CI, 9.8%-16.1%) were being treated with stimulants, 2.3% (95% CI, 1.3%-4.3%) with antidepressants, and 1.9% (95% CI, 1.0%-3.7%) with clonidine. Among children without ADHD, 0.5% (95% CI, 0.3%-0.8%) were receiving stimulant medication. This represented 22.9% (95% CI, 14.6%-34.0%) of all the children who were receiving stimulants. Variables significantly associated with stimulant use were being male, having ADHD, attending a paediatrician, and having higher scores on the Aggressive Behaviour and Attention Problems scales on the Child Behaviour Checklist. CONCLUSIONS: About 13% of Australian children with ADHD, and a substantial number of children without ADHD, are taking stimulants. The question of whether Australian children are being undertreated or overtreated with stimulant medication depends on the criteria used to assess the appropriateness of stimulant use. Additional information is needed to clarify when stimulants should be used to treat ADHD. PMID- 12088475 TI - Guideline-discordant care in acute myocardial infarction: predictors and outcomes. AB - OBJECTIVES: To determine (i) factors which predict whether patients hospitalised with acute myocardial infarction (AMI) receive care discordant with recommendations of clinical practice guidelines; and (ii) whether such discordant care results in worse outcomes compared with receiving guideline-concordant care. DESIGN: Retrospective cohort study. SETTING: Two community general hospitals. PARTICIPANTS: 607 consecutive patients admitted with AMI between July 1997 and December 2000. MAIN OUTCOME MEASURES: Clinical predictors of discordant care; crude and risk-adjusted rates of inhospital mortality and reinfarction, and mean length of hospital stay. RESULTS: At least one treatment recommendation for AMI was applicable for 602 of the 607 patients. Of these patients, 411(68%) received concordant care, and 191 (32%) discordant care. Positive predictors at presentation of discordant care were age > 65 years (odds ratio [OR], 2.5; 95% CI, 1.7-3.6), silent infarction (OR, 2.7; 95% CI, 1.6-4.6), anterior infarction (OR, 2.5; 95% CI, 1.7-3.8), a history of heart failure (OR, 6.3; 95% CI, 3.7 10.7), chronic atrial fibrillation (OR, 3.2; 95% CI, 1.5-6.4); and heart rate >/= 100 beats/min (OR, 2.1; 95% CI, 1.4-3.1). Death occurred in 12.0% (23/191) of discordant-care patients versus 4.6% (19/411) of concordant-care patients (adjusted OR, 2.42; 95% CI, 1.22-4.82). Mortality was inversely related to the level of guideline concordance (P = 0.03). Reinfarction rates also tended to be higher in the discordant-care group (4.2% v 1.7%; adjusted OR, 2.5; 95% CI, 0.90 7.1). CONCLUSIONS: Certain clinical features at presentation predict a higher likelihood of guideline-discordant care in patients presenting with AMI. Such care appears to increase the risk of inhospital death. PMID- 12088476 TI - A quality use of medicines program for continuity of care in therapeutics from hospital to community. AB - OBJECTIVE: To evaluate adherence to an agreed minimum dataset for patient medication information exchange between hospitals and general practitioners. DESIGN: Quasi pre-post design with a supplementary post-program comparison group; sequential descriptive surveys of patients following recent discharge from public hospitals; opinion questionnaire for key hospital informants; stakeholder forum to review data and Quality Use of Medicines (QUM) action plans between pre- and post-program surveys. PARTICIPANTS AND SETTING: GPs and hospitals from the South East Area Health Service, Sydney. During 2000-2001, 81/124 GPs (65%) returned 147 patient questionnaires in initial follow-up; 88/119 GPs (74%) returned 131 questionnaires in final follow-up; a supplementary group of 54/120 GPs (45%) returned 66 questionnaires; 32/45 (71%) of nominated key informants responded to the hospital survey. RESULTS: Direct notification of GPs by hospitals of their patient's admission was unchanged from the initial level of 22%. The proportion of GPs providing medication information to the hospital increased from 38% to 51% at Stage 2 (P < 0.05) and remained at 52% at Stage 3. The proportion of GPs receiving discharge summaries directly (initially 2%) increased to 26% at Stage 2 (P < 0.001) and remained at 27% at Stage 3. CONCLUSIONS: System change is slow to occur, but changes that are implemented are maintained. The stakeholder forum suggested that a specific person should be responsible for GP liaison. PMID- 12088477 TI - Recommendations for lightning protection in sport. AB - Lightning is an important cause of weather-related morbidity and mortality and is often underrated. There has been an increase in the proportion of casualties occurring during outdoor sport and recreational activities over recent years. However, in Australia, there is a deficiency in recommendations for lightning safety at sporting events. Organisers of sporting events should have a lightning safety policy that includes a designated weather watcher with the authority to stop or postpone the event, a specific chain of command, and designated safe areas. Suspension and resumption of play should follow the "30/30" rule: play should stop when the flash-to-bang count is 30 seconds, and should not resume until 30 minutes after the last lightning. At events with large crowds, additional time should be allowed for evacuating all people to safe areas. PMID- 12088478 TI - Australia needs to follow New Zealand's lead on sports injuries. AB - Sports injuries result in substantial costs to the Australian community and also act as a barrier to increased participation in physical activity. However, the Australian healthcare system has no coordinated approach for monitoring or preventing sports injuries. This is in contrast to New Zealand, which has a specific body responsible for managing sports injuries, in a similar way to work injuries and traffic accidents. PMID- 12088479 TI - Clinical trials without consent: some experiments simply cannot be done. PMID- 12088480 TI - Transient cortical blindness related to coronary angiography and graft study. PMID- 12088481 TI - Emerging viral infections in Australia. AB - Hendra virus infection should be suspected in someone with close association with horses or bats who presents acutely with pneumonia or encephalitis (potentially after a prolonged incubation period). Australian bat lyssavirus infection should be suspected in a patient with a progressive neurological illness and a history of exposure to a bat. Rabies vaccine and immunoglobulin should be strongly considered after a bite, scratch or mucous membrane exposure to a bat. Japanese encephalitis vaccine should be considered for people intending to reside in or visit endemic areas of southern or eastern Asia for more than 30 days. PMID- 12088482 TI - Medical indemnity. PMID- 12088483 TI - Chronic fatigue syndrome clinical practice guidelines. PMID- 12088484 TI - Howard Florey, Alexander Fleming and the fairy tale of penicillin. PMID- 12088485 TI - Hyponatraemia and hypokalaemia caused by indapamide. PMID- 12088486 TI - End-of-life issues. PMID- 12088487 TI - Community-acquired MRSA bacteraemia. PMID- 12088489 TI - Conference overview: a duty of care. PMID- 12088490 TI - The student and junior doctor in distress. PMID- 12088491 TI - A student mental health and welfare program in a medical faculty. PMID- 12088492 TI - Stress in a graduate medical degree. PMID- 12088493 TI - Tasmanian University Medical Students Society student mentor scheme: a model to help students in distress. PMID- 12088494 TI - Examining stress and responses to stress in medical students and new medical graduates. PMID- 12088495 TI - Registration of medical students by medical boards. PMID- 12088496 TI - Overview: the experience of the Health Committee of the South Australian Medical Board. PMID- 12088497 TI - Attitudes to healthcare and self-care among junior medical officers: a preliminary report. PMID- 12088498 TI - The junior doctor in distress: the role of a medical education officer at the systems level. PMID- 12088499 TI - The junior doctor in distress: the role of a medical education officer at the individual level. PMID- 12088500 TI - Overview: the experience of the New South Wales Medical Board. PMID- 12088501 TI - Changes to the South Australian Medical Practitioners Act 1983. PMID- 12088502 TI - Return to work for junior doctors after ill-health. PMID- 12088503 TI - Reports from breakout groups. PMID- 12088504 TI - Oligomerization status, with the monomer as active species, defines catalytic efficiency of UDP-glucose pyrophosphorylase. AB - Barley UDP-glucose pyrophosphorylase (UGPase), a key enzyme for the synthesis of sucrose, cellulose and other saccharides, was expressed in Escherichia coli and purified. Using both native electrophoresis and gel filtration, the recombinant and crude leaf UGPase proteins were found to exist as a mixture of monomers, dimers and higher-order polymers. In order to understand the molecular basis for the oligomerization of UGPase, a conserved Cys residue was replaced (C99S mutant) and several amino acids were substituted (LIV to NIN, KK to LL and LLL to NNN) in a conserved hydrophobic domain (amino acids 117-138). The C99S mutant had about half the V (max) of the wild-type and a 12-fold higher K (m) for PP(i), whereas NIN and LL mutations lowered the V (max) by 12- and 2-fold, respectively, with relatively small effects on substrate K (m) values (the NNN mutant was insoluble/inactive). The NIN mutation resulted in a low-activity oligomerized enzyme form, with very little monomer formation. Activity staining on native PAGE gels as well as gel-filtration studies demonstrated that the monomer was the sole enzymically active form. Possible implications of the oligomerization status of UGPase for post-translational regulation of the enzyme are discussed. PMID- 12088505 TI - New insights into long-chain fatty acid uptake by heart muscle: a crucial role for fatty acid translocase/CD36. AB - Long-chain fatty acids are an important source of energy for several cell types, in particular for the heart muscle cell. Three different proteins, fatty acid translocase (FAT)/CD36, fatty acid transport protein and plasma membrane fatty acid binding protein, have been identified as possible membrane fatty acid transporters. Much information has been accumulated recently about the fatty acid transporting function of FAT/CD36. Several experimental models to study the influence of altered FAT/CD36 expression on fatty acid homoeostasis have been identified or developed, and underscore the importance of FAT/CD36 for adequate fatty acid transport. These models include the FAT/CD36 null mouse, the spontaneously hypertensive rat and FAT/CD36-deficient humans. The fatty acid transporting role of FAT/CD36 is further demonstrated in mice overexpressing muscle-specific FAT/CD36, and in transgenic mice generated using a genetic-rescue approach. In addition, a wealth of information has been gathered about the mechanisms that regulate FAT/CD36 gene expression and the presence of functional FAT/CD36 on the plasma membrane. Available data also indicate that FAT/CD36 may have an important role in the aetiology of cardiac disease, especially cardiac hypertrophy and diabetic cardiomyopathy. This review discusses our current knowledge of the three candidate fatty acid transporters, the metabolic consequences of alterations in FAT/CD36 levels in different models, and the mechanisms that have been identified for FAT/CD36 regulation. PMID- 12088506 TI - Determinants of adenophostin A binding to inositol trisphosphate receptors. AB - Inositol 1,4,5-trisphosphate (IP(3)) receptors from cerebellum and recombinant type 1 IP(3) receptors expressed in Sf9 cells had indistinguishable affinities for IP(3) ( K (d)=6.40+/-0.48 nM) and adenophostin A ( K (d)=0.89+/-0.05 nM). In cytosol-like medium, each of the three mammalian IP(3) receptor subtypes when expressed in Sf9 cells bound adenophostin A with greater affinity than IP(3). It has been suggested that adenophostin A binds with high affinity only in the presence of ATP, but we found that adenophostin A similarly displaced [(3)H]IP(3) from type 1 IP(3) receptors whatever the ATP concentration. N-terminal fragments of the type 1 receptor were expressed with and without the S1 splice site; its removal had no effect on [(3)H]IP(3) binding to the 1-604 protein, but abolished binding to the 224-604 protein. The 1-604 fragment and full-length receptor bound adenophostin A with the same affinity, but the fragment had 3-fold greater affinity for IP(3), suggesting that C-terminal residues selectively inhibit IP(3) binding. The 224-604S1(+) fragment bound IP(3) and adenophostin A with increased affinity, but as with the 1-604 fragment it bound adenophostin A with only 2-fold greater affinity than IP(3). High-affinity binding of adenophostin A may be partially determined by its 2'-phosphate interacting more effectively than the 1 phosphate of IP(3) with residues within the IP(3)-binding core. This may account for the 2-fold greater affinity of adenophostin A relative to IP(3) for the minimal IP(3)-binding domain. In addition we suggest that C-terminal residues, which impede access of IP(3), may selectively interact with adenophostin A to allow it unhindered access to the IP(3)-binding domain. PMID- 12088508 TI - Non-essential roles of cysteine residues in functional expression and redox regulatory pathways for canine glutamate/aspartate transporter based on mutagenic analysis. AB - A redox regulatory mechanism and a molecular link between oxidative and excitotoxic neurodegeneration have been postulated for high-affinity Na(+) dependent glutamate transporters. In the present study, mutations were introduced at three cysteine residues in canine glutamate/aspartate transporter (GLAST) to investigate the functional significance of thiol groups in response to oxidation. Cys(-) GLAST, in which all cysteines were replaced by other amino acids, as well as other mutants with disruption of one of three cysteine residues, showed insoluble oligomer formation, which was considered to be due to spontaneous and excessive oxidation as observed in wild-type GLAST. The mutant transporters also showed plasma-membrane localization and glutamate-transport kinetics that were very similar to those of wild-type GLAST. Glutamate-transport activities in COS-7 cells transfected with wild-type and Cys(-) GLAST were inhibited to the same degree when cells were exposed to Hg(2+) and were recovered by the addition of thiol-specific reductant dithiothreitol. These findings suggest that cysteine residues are not critical in functional expression of GLAST and the redox-sensing pathway via glutamate transporters. PMID- 12088507 TI - Measurement of free and membrane-bound cathepsin G in human neutrophils using new sensitive fluorogenic substrates. AB - Activated human polymorphonuclear neutrophils at inflammatory sites release the chymotrypsin-like protease cathepsin G, together with elastase and proteinase 3 (myeloblastin), from their azurophil granules. The low activity of cathepsin G on synthetic substrates seriously impairs studies designed to clarify its role in tissue inflammation. We have solved this problem by producing new peptide substrates with intramolecularly quenched fluorescence. These substrates were deduced from the sequence of putative protein targets of cathepsin G, including the reactive loop sequence of serpin inhibitors and the N-terminal domain of the protease-activated receptor of thrombin, PAR-1. Two substrates were selected, Abz TPFSGQ-EDDnp and Abz-EPFWEDQ-EDDnp, that are cleaved very efficiently by cathepsin G but not by neutrophil elastase or proteinase 3, with specificity constants (k(cat)/K(m)) in the 10(5) M(-1).s(-1) range. They can be used to measure subnanomolar concentrations of free enzyme in vitro and at the surface of neutrophils purified from fresh human blood. Purified neutrophils express 0.02 0.7 pg of cathepsin G/cell (n=15) at their surface. This means that about 10(4) purified cells may be enough to record cathepsin G activity within minutes. This may be most important for investigating the role of cathepsin G as an inflammatory agent, especially in bronchoalveolar lavage fluids from patients with pulmonary inflammatory disorders. PMID- 12088509 TI - Pulse pressure and age at menopause. AB - BACKGROUND: The objective of this study was to study the association of early age at menopause with pulse pressure (PP), a marker of arterial stiffness, and PP change. METHODS: The effect of natural menopause was studied in 2484 women from the Atherosclerosis Risk in Communities (ARIC) Study who had not used hormone replacement therapy and who had not had a hysterectomy. The cross-sectional association of age with PP was evaluated in the entire cohort. The cross sectional association of recalled age at menopause was evaluated in the 1688 women who were postmenopausal at baseline. PP change over 6 years was assessed in relation to menopausal age separately in women who were postmenopausal at baseline and in those whose menopause occurred during the 6-year interval. RESULTS: Chronological age was strongly and positively associated with PP in cross-sectional analyses, but not independently associated with PP change. While menopausal age was not associated cross-sectionally with PP, early age at menopause (age<45) was significantly and independently associated with a slightly larger increase in PP (8.4, 95% CI 7.0-9.8) than later menopause (6.5, 95% CI 5.8;7.2). However, among normotensive women the difference was not statistically significant (p = 0.07, 6.1 vs 4.7). CONCLUSIONS: Early age at menopause may be related to a greater increase in arterial stiffness, but the effect appears to be small and further evidence is needed. PMID- 12088510 TI - Functional foods: concepts and application to inulin and oligofructose. AB - A food can be regarded as functional if it is satisfactorily demonstrated to affect beneficially one or more target functions in the body, beyond adequate nutritional effects, in a way which is relevant to either the state of well-being and health or the reduction of the risk of a disease. Health claims are expected to be authorized for functional foods based either on enhanced function (type A claim) or disease risk reduction (type B claim). Their development is a unique opportunity to contribute to the improvement of the quality of the food offered to consumer's choice for the benefit of his well-being and health. But only a rigorous scientific approach producing sound data will guarantee its success. The functional food components that are discussed in the proceedings of the 3rd ORAFTI Research Conference are the inulin-type fructans, natural food components found in miscellaneous edible plants. They are non-digestible oligosaccharides that are classified as dietary fiber. The targets for their functional effects are the colonic microflora that use them as selective 'fertilizers'; the gastrointestinal physiology; the immune functions; the bioavailability of minerals; and the metabolism of lipids. Potential health benefits may also concern reduction of the risk of some diseases like intestinal infections, constipation, non-insulin dependent diabetes, obesity, osteoporosis or colon cancer. The present proceedings review the scientific data available and, by reference to the concepts in functional food science, they assess the scientific evidence which will be used to substantiate health claims. PMID- 12088511 TI - Gastrointestinal effects of prebiotics. AB - The defining effect of prebiotics is to stimulate selectively the growth of bifidobacteria and lactobacilli in the gut and, thereby, increase the body's natural resistance to invading pathogens. Prebiotic carbohydrates may also have additional, less specific, benefits because they are fermented in the large intestine. The prebiotic carbohydrates that have been evaluated in humans at the present time largely consist of fructans or galactans. There is consistent evidence from in vitro and in vivo studies that these are not digested by normal human enzymes, but are readily fermented by anaerobic bacteria in the large intestine. There are no reports of faecal recovery of measurable quantities of prebiotic carbohydrates. Through fermentation in the large intestine, prebiotic carbohydrates yield short-chain fatty acids, stimulate the growth of many bacterial species in addition to the selective effects on lactobacilli and bifidobacteria, they can also produce gas. Along with other fermented carbohydrates, prebiotics have mild laxative effects, although this has proved difficult to demonstrate in human studies because the magnitude of laxation is small. Potentially, the most important effect of prebiotic carbohydrates is to strengthen the body's resistance to invading pathogens and, thereby, prevent episodes of diarrhoea. At the present time, this effect has not been convincingly demonstrated in either adults or children, although there have been attempts to ameliorate the diarrhoea associated with antibiotics and travel, but without success. However, prebiotic carbohydrates clearly have significant and distinctive physiological effects in the human large intestine, and on the basis of this it is likely that they will ultimately be shown to be beneficial to health. PMID- 12088512 TI - Nutritional advantages of probiotics and prebiotics. AB - The potential 'nutritional advantages' of probiotics and prebiotics consist of preventive, and sometimes curative, effects against certain diseases. The evidence supporting such advantages, which requires randomised controlled trials and consistency of results from study to study, is rapidly increasing. This article summarizes the effects against diseases of intestinal origin. There is a high level of evidence for positive effects of some prebiotics to alleviate constipation and treat hepatic encephalopathy. Interesting aspects, but with a lower level of evidence at the present time, include prevention of colon cancer, intestinal infection, and recurrence of inflammatory bowel disease. There is a high level of evidence for positive effects of some probiotics in the alleviation of lactose intolerance, antibiotic-associated intestinal disorders and gastroenteritis. Evidence is rapidly growing regarding the prevention of recurrence of inflammatory bowel diseases. Positive trials have suggested preventive effects against intestinal colonization with specific gut pathogens including Clostridium difficile and Helicobacter pylori. PMID- 12088513 TI - Inulin and oligofructose in the dietary fibre concept. AB - Dietary fibre consists of remnants of edible plant cell polysaccharides and associated substances resistant to hydrolysis by human alimentary enzymes, which may benefit health through a wide range of physiological effects. Inulin and oligofructose are storage carbohydrates found in a number of vegetables, fruits and whole grains. They resist digestion and absorption in the stomach and small intestine of humans, as shown by their almost full recovery at the end of the ileum of healthy or ileostomised volunteers. Inulin and oligofructose thus enter into the large intestine where they are available to fermentation, as demonstrated by increased breath hydrogen. Fermentation of both substrates is complete and no residue is found in human stools. Inulin and oligofructose improve laxation. Their bulking capacity comprised between 1.2 and 2.1 g of stool per g of ingested substrate, results mainly from increases in microbial biomass in the colon. As water content of bacterial cells is high, stools are softer and easier to expulse. Stool frequency is thus increased, particularly in slightly constipated individuals. In addition, likely due to their fermentation properties, inulin and oligofructose also affect the intestinal epithelium (trophicity, mucin expression, etc.), that may strengthen mucosal protection and reduce the risk of gastrointestinal diseases. In summary, inulin and oligofructose are plant carbohydrates, resistant to digestion in the human small intestine and fermented by colonic bacteria. They exert several intestinal physiological effects contributing to maintenance of health. Therefore, inulin and oligofructose fit well within the current concept of dietary fibre. PMID- 12088514 TI - Fermentation and bulking capacity of indigestible carbohydrates: the case of inulin and oligofructose. AB - The bulking index (i.e. the increase in faecal fresh weight in gram per gram indigestible carbohydrate ingested) with oligofructose and inulin is similar to that produced with other easily fermented fibres such as pectins and gums. Most studies in man have been performed at a level of 15 g/d and more investigations on lower intakes are needed to appoint the least intake for an effect. Concerning short-chain fatty acids (SCFA) most studies have been using oligofructose and points at an increased butyric acid formation in the caecum of rats. In one study on rats with inulin high caecal proportions of propionic acid were obtained. As inulin has a higher molecular weight than oligofructose it might be speculated if this could be a reason to the different SCFA-profile formed. No effects on faecal concentrations of SCFA in humans have been revealed with inulin and oligofructose, which neither is expected as most of the SCFA formed during the fermentation already has been absorbed or utilized by the colonic mucosa. PMID- 12088515 TI - Calcium intake, calcium bioavailability and bone health. AB - Calcium accounts for 1-2 % of adult human body weight. Over 99 % of total body Ca is found in the teeth and bones. Therefore, in addition to the obvious structural role of the skeleton, it also serves as a reservoir for Ca. Dietary Ca intake has an important impact on bone metabolism and bone health. Chronic Ca deficiency resulting from inadequate intake or poor intestinal absorption is one of several important causes of reduced bone mass and osteoporosis. It is vital, therefore, that adequate dietary Ca is consumed at all stages of life - in early life so that the genetically programmed peak bone mass can be reached and in later adulthood so that the skeletal mass can be maintained and age-related bone loss minimised. Unfortunately, there is wide variation in the estimates of daily Ca requirements made by different expert authorities. Furthermore, there is evidence that many individuals are not consuming these recommended levels. The consequence of this for bone health will be discussed in the present review. Besides the amount of Ca in the diet, the absorption of dietary Ca in foods is also a critical factor in determining the availability of Ca for bone development and maintenance. Thus, there is a need to identify food components and/or functional food ingredients that may positively influence Ca absorption in order to ensure that Ca bioavailability from foods can be optimised. This approach may be of particular value in individuals who fail to achieve the dietary recommended level of Ca. PMID- 12088516 TI - Inulin, oligofructose and mineral metabolism - experimental data and mechanism. AB - Numerous investigations performed in animal models in the past 10 years have shown repeatedly that non-digestible oligosaccharides (NDO), such as inulin, oligofructose or transgalacto-oligosaccharides (TOS), stimulate mineral absorption, mainly calcium and magnesium. Long-term beneficial effects on bone health have been indicated by accumulation of bone mineral content in growing rats or prevention of bone loss in ovariectomized rats. However, bone mineral content or density are not necessarily associated with bone quality. In recent studies both oligofructose and calcium prevented loss of trabecular bone area induced by oestrogen deficiency, this, however, occurred at different trabecular shapes. The effects of NDO on mineral metabolism may be based on the enhancement of passive and active mineral transport across the intestinal epithelium, mediated by an increase in certain metabolites of the intestinal flora and a reduction of pH. The possible impact of short-chain fatty acids, butyrate in particular, and of polyamines on the stimulation of mineral absorption capacity, and the interaction of oligofructose and antibiotics is discussed. PMID- 12088517 TI - Non-digestible oligosaccharides and calcium absorption in girls with adequate calcium intakes. AB - Non-digestible oligosaccharides such as inulin and oligofructose have been shown to consistently increase calcium absorption in experimental animals, but data in humans are less clear-cut. The objective of this study was to assess the effect of 8 g/d of oligofructose or a mixture of inulin and oligofructose on calcium absorption in girls at or near menarche. A total of fifty-nine subjects were studied using a balanced, randomized, cross-over design. They received, in random order, 8 g/d placebo (sucrose), oligofructose or the mixture inulin+oligofructose for 3 weeks, separated by a 2-week washout period. Throughout the study, subjects consumed a total of approximately 1500 mg/d dietary calcium, by adding two glasses of calcium-fortified orange juice to their diet. Four grams of placebo, oligofructose or the mixture inulin+oligofructose was added to each glass of orange juice immediately before it was consumed. At the end of each 3-week adaptation period, calcium absorption was measured, using a dual stable isotope technique, from the cumulative fractional excretion of an oral and an intravenous tracer over 48 hours. Calcium absorption was significantly higher in the group receiving the inulin+oligofructose mixture than in the placebo group v. P=0.01), but no significant difference was seen between the oligofructose group and the placebo group v. P=NS). We conclude that modest intakes of an inulin+oligofructose mixture increases calcium absorption in girls at or near menarche. PMID- 12088518 TI - Prebiotic effects of inulin and oligofructose. AB - Prebiotics are non-digestible food ingredients that target certain components within the microbiota of the human large intestine. Efficient prebiotics need to have a specific fermentation therein and thereby have the ability to alter the faecal microflora composition towards a more 'beneficial' community structure. This should occur by the stimulation of benign or potentially health promoting genera but not the harmful groups. Because of their positive attributes bifidobacteria and lactobacilli are the most frequent target organisms. Both inulin and oligofructose have been demonstrated to be effective prebiotics. This has been shown through both in vitro and in vivo assessments in different laboratories. Because of their recognised prebiotic properties, principally the selective stimulation of colonic bifidobacteria, both inulin and oligofructose are increasingly used in new food product developments. Examples include drinks, yoghurts, biscuits and table spreads. Because of the recognised inhibitory effects that bifidobacteria can exert against gut pathogens, one of the most important aspects of prebiotic ingestion is fortification of the gut flora to resist acute infections. PMID- 12088519 TI - Molecular methods for exploring the intestinal ecosystem. AB - Molecular methods have provided renewed impetus for the analysis of the composition of the intestinal microflora in health and disease. The polymerase chain reaction coupled with denaturing gradient gel electrophoresis provides a method whereby the bacterial communities in large numbers of samples can be compared efficiently and effectively. Altered bacterial populations associated with disease states can then be targeted for further investigation. In the long term, an 'abnormal microflora' might be rectified by the use of probiotics or prebiotics. PMID- 12088520 TI - Molecular biological methods for studying the gut microbiota: the EU human gut flora project. AB - Seven European laboratories co-operated in a joint project (FAIR CT97-3035) to develop, refine and apply molecular methods towards facilitating elucidation of the complex composition of the human intestinal microflora and to devise robust methodologies for monitoring the gut flora in response to diet. An extensive database of 16S rRNA sequences for tracking intestinal bacteria was generated by sequencing the 16S rRNA genes of new faecal isolates and of clones obtained by amplification with polymerase chain reaction (PCR) on faecal DNA from subjects belonging to different age groups. The analyses indicated that the number of different species (diversity) present in the human gut increased with age. The sequence information generated, provided the basis for design of 16S rRNA directed oligonucleotide probes to specifically detect bacteria at various levels of phylogenetic hierarchy. The probes were tested for their specificity and used in whole-cell and dot-blot hybridisations. The applicability of the developed methods was demonstrated in several studies and the major outcomes are described. PMID- 12088521 TI - Oligofructose and experimental model of neonatal necrotising enterocolitis. AB - The gut of preterm neonates is colonised with a paucity of bacterial species originating more from the environment than from the mother. Furthermore, a delayed colonisation by bifidobacteria promotes colonisation by potentially pathogenic bacteria. This may contribute towards the development of neonatal necrotising enterocolitis (NEC). The physiopathology of NEC is still unclear but immaturity of the gut, enteral feeding and bacterial colonisation are all thought to be involved. None of the current preventive treatments are considered satisfactory. Modulating the autochthonous microflora by probiotics or prebiotics could be a more reliable approach to prevention. Using gnotobiotic quails as an experimental model of NEC we have shown that onset of intestinal lesions requires a combination of low endogenous lactase activity, lactose in diet, and colonisation by lactose-fermenting bacteria such as the clostridia. The protective role of bifidobacteria was demonstrated in this model through a decrease in clostridial populations and in butyric acid. Oligofructose dietary supplementation was shown to enhance this effect with an increase in the bifidobacterial level and consequently a greater decrease in clostridia. However, oligofructose was unable to promote a bifidobacterial acquisition when the microflora was initially deprived of this group. Nevertheless, oligofructose can act as an anti-infective agent and decrease the occurrence or severity of the lesions depending on the bacteria involved. According to these results and to the fact that oligosaccharides are a major component of breast milk, the addition of oligofructose in formula milks may be a nutritional approach to favouring colonisation by a beneficial flora. PMID- 12088522 TI - The immune-enhancing effects of dietary fibres and prebiotics. AB - The gastrointestinal tract is subjected to enormous and continual foreign antigenic stimuli from food and microbes. This organ must integrate complex interactions among diet, external pathogens, and local immunological and non immunological processes. It is critical that protective immune responses are made to potential pathogens, while hypersensitivity reactions to dietary antigens are minimised. There is increasing evidence that fermentable dietary fibres and the newly described prebiotics can modulate various properties of the immune system, including those of the gut-associated lymphoid tissues (GALT). This paper reviews evidence for the immune-enhancing effects of dietary fibres. Changes in the intestinal microflora that occur with the consumption of prebiotic fibres may potentially mediate immune changes via: the direct contact of lactic acid bacteria or bacterial products (cell wall or cytoplasmic components) with immune cells in the intestine; the production of short-chain fatty acids from fibre fermentation; or by changes in mucin production. Although further work is needed to better define the changes, mechanisms for immunomodulation, and the ultimate impact on immune health, there is convincing preliminary data to suggest that the consumption of prebiotics can modulate immune parameters in GALT, secondary lymphoid tissues and peripheral circulation. Future protocols on the physiological impact of consuming prebiotics should be designed to include assessments of the gut microflora, gut physiology and the function and composition of the various regions of GALT. PMID- 12088523 TI - Non-digestible oligosaccharides and defense functions: lessons learned from animal models. AB - Animals are constantly exposed to a diversity of health challenges and the gastrointestinal tract (GIT) is a major, if not the principal, site of exposure. Animal models and a limited number of human clinical studies have shown that the assemblages and metabolic activities of the resident bacteria are important determinants of the effectiveness of the various host defense mechanisms and thereby influence the ability of animals to respond to health challenges. The assemblages of bacteria resident in the GIT provide a first line of defense that can exclude invading pathogens, reduce the proliferation of opportunistic pathogens already resident in the GIT, and reduce the availability, carcinogenicity, or toxicity of noxious chemicals. The mucosa of the GIT is a second, multilayered line of defense that includes the mucous and other secretions, the epithelial cells, and immune-associated cells scattered within and under the epithelium. The final line of defense contends with pathogens or noxious chemicals that transcend the mucosal barrier and enter the host and consists of the innate and acquired components of the systemic immune system and the xenobiotic metabolizing enzymes. The lactic acid producing bacteria (LAB) are considered to be immunomodulatory and directly or indirectly influence the GIT and systemic defense functions. Corresponding with this, supplementing the diet with inulin, oligofructose, or other nondigestible oligosaccharides that increase the densities and metabolic capacities of the LAB enhances defense mechanisms of the host, increases resistance to various health challenges, and accelerates recovery of the GIT after disturbances. PMID- 12088524 TI - Human studies with probiotics and prebiotics: clinical implications. AB - Probiotic agents have been shown to have significant clinical beneficial effects in the prevention and management of gastrointestinal and non-gastrointestinal conditions. These observations have led to work demonstrating that an important mechanism of these agents is their close interaction with the gut associated lymphoid tissue (GALT) and suggested immunomodulatory effects on systemic immune response. Studies on the possibility that prebiotic agents might directly or indirectly induce similar immunomodulation have only recently begun. The preliminary findings of several recent human clinical trials reviewed in this article indicate that prebiotics may indeed prove to be a clinically beneficial dietary supplement, in the context of novel nutritional strategies for the management of gastrointestinal and systemic conditions. PMID- 12088525 TI - Dietary carbohydrate's effects on lipogenesis and the relationship of lipogenesis to blood insulin and glucose concentrations. AB - The process by which dietary carbohydrate is transformed into fat in the human body is termed de novo lipogenesis. New methods for the measurement of this process in humans are available and have been used to investigate the role of the carbohydrate form (fed as a liquid or solid), the level of processing of carbohydrate in foods, and the role of lipogenesis in the control of liver triacylglycerol secretion. The present paper will discuss how research results are affected by both the physical state of the carbohydrate in the diet and by the metabolic state of individual research subjects. Of interest is the relationship between the glycemic index of a food (or indicators of a food's glycemic index) and that food's ability to stimulate lipogenesis in humans. Given the increasing prevalence of obesity worldwide, future scientific emphasis will expand methods to quantitate the lipogenic potential of specific foods and dietary patterns and investigate how the metabolic state of insulin resistance affects lipogenesis and/or contributes to obesity. PMID- 12088526 TI - Inulin and oligofructose modulate lipid metabolism in animals: review of biochemical events and future prospects. AB - Inulin and oligofructose, besides their effect on the gastro-intestinal tract, are also able to exert 'systemic' effect, namely by modifying the hepatic metabolism of lipids in several animal models. Feeding male Wistar rats on a carbohydrate-rich diet containing 10 % inulin or oligofructose significantly lowers serum triacylglycerol (TAG) and phospholipid concentrations. A lower hepatic lipogenesis, through a coordinate reduction of the activity and mRNA of lipogenic enzymes is a key event in the reduction of very low-density lipoprotein TAG secretion by oligofructose. Oligofructose is also able to counteract triglyceride metabolism disorder occurring through dietary manipulation in animals, and sometimes independently on lipogenesis modulation: oligofructose reduces post-prandial triglyceridemia by 50 % and avoids the increase in serum free cholesterol level occurring in rats fed a Western-type high fat diet. Oligofructose protects rats against liver TAG accumulation (steatosis) induced by fructose, or occurring in obese Zucker fa/fa rats. The protective effect of dietary inulin and oligofructose on steatosis in animals, would be interesting, if confirmed in humans, since steatosis is one of the most frequent liver disorders, occurring together with the plurimetabolic syndrome, in overweight people. The panel of putative mediators of the systemic effects of inulin and oligofructose consists in either modifications in glucose/insulin homeostasis, the end-products of their colonic fermentation (i.e. propionate) reaching the liver by the portal vein, incretins and/or the availability of other nutrients. The identification of the key mediators of the systemic effects of inulin and oligofructose is the key to identify target function(s) (or dysfunction(s)), and finally individuals who would take an advantage of increasing their dietary intake. PMID- 12088527 TI - Inulin and oligofructose: effects on lipid metabolism from human studies. AB - Although convincing lipid-lowering effects of the fructo-oligosaccharide, inulin, have been demonstrated in animals, attempts to reproduce similar effects in man have produced conflicting findings. This may be because of the much lower doses which can be used due to the adverse gastrointestinal symptoms exhibited by most subjects consuming in excess of 15 g/d. There are nine studies reported in the literature which have investigated the response of blood lipids (usually total and LDL-cholesterol and triacylglycerol) to inulin or oligofructose supplementation in human volunteers. Three have observed no effects of inulin or oligofructose on blood levels of cholesterol or triacylglycerol, three have shown significant reductions in triacylglycerol, whilst four have shown modest reductions in total and LDL-cholesterol. Studies have been conducted in both normo- and moderately hyperlipidaemic subjects. Differences in study outcomes do not appear to be due to differences in the type or dose of oligosaccharides used nor the duration of the studies. Because animal studies have identified inhibition of hepatic fatty acid synthesis as the major site of action for the triacylglycerol lowering effects of inulin and oligofructose, and because this pathway is relatively inactive in man unless a high carbohydrate diet is fed, variability in response may be a reflection of differences in background diet or the experimental foods used. PMID- 12088528 TI - Strategies for cancer prevention: the role of diet. AB - Linkages between diet habits and cancer risk have surfaced from a multitude of epidemiological and preclinical studies. Collectively these studies provide rather compelling evidence that dietary components modify the incidence and biological behavior of tumors. While the risk of breast, prostate, colon, lung and liver cancers are frequently associated with dietary patterns, inconsistencies are not uncommon. These inconsistencies likely reflect the multi factorial and complex nature of cancer and the specificity that individual dietary constituents have in modifying cancer related genetic pathways. The complexity of defining the role of diet is underscored by the numerous and diverse essential and non-essential components that may alter one or more phases of the cancer process. The explosive increase in the recognition of genes and pathways for regulating cell growth and development, and evaluating the response to hormones and other chemicals synthesized by the body, offers exciting opportunities for unraveling the molecular targets by which dietary components influence cancer prevention. It is recognized that all cells have unique 'signatures' that are characterized by active and inactive genes and cellular products. It is certainly plausible that bridging knowledge about these unique cellular characteristics with the molecular targets for nutrients can be used to assist in optimizing nutrition and minimizing cancer risk. PMID- 12088529 TI - Experimental evidences on the potential of prebiotic fructans to reduce the risk of colon cancer. AB - Inulin is extracted from the chicory root. It is a set of fructans with its monomers linked by means of beta(2-1) bonds. This linkage cannot be hydrolysed by either pancreatic or by brush border digestive enzymes in the upper intestinal tract of humans. As such the carbohydrates arrive in the colon, where they are fermented by bifidobacteria and other lactic acid producing bacteria, thus enhancing their relative populations in the gut. Recent research in experimental animal models revealed that inulin has significant anticarcinogenic properties. It acts chemopreventively by reducing the incidence of azoxymethane (AOM) - induced aberrant crypt foci and tumours in the colon. These effects may be due to the stimulation of bifidobacteria, which themselves have been shown to act as antigenotoxic in the colon and to reduce AOM-induced tumours. Also fermentation products, including the short-chain fatty acid butyrate, could contribute to the protective effects. In this case a mechanism may be the induction of apoptosis of already transformed cells. The experimental evidence from animal studies and from studies elucidating potential mechanisms strongly supports the possibility that inulin will contribute to reducing risks for colon cancer in humans. In order to obtain more insight into this possibility, human dietary intervention studies relating biomarkers of reduced risk to inulin consumption are needed. PMID- 12088530 TI - Inulin/oligofructose and anticancer therapy. AB - The results of our investigations indicate that dietary treatment with inulin or oligofructose incorporated in the basal diet for experimental animals: (i) reduced the incidence of mammary tumors induced in Sprague-Dawley rats by methylnitrosourea; (ii) inhibited the growth of transplantable malignant tumors in mice; and (iii) decreased the incidence of lung metastases of a malignant tumor implanted intramuscularily in mice. Moreover, besides such cancer risk reduction effects, the dietary treatment with inulin or oligofructose significantly potentiated the effects of subtherapeutic doses of six different cytotoxic drugs commonly utilized in human cancer treatment. If confirmed, such dietary treatment with inulin or oligofructose potentiating cancer therapy might become an interesting approach to complement classical protocols of human cancer treatment without any additional risk for the patients. PMID- 12088531 TI - Technological functionality of inulin and oligofructose. AB - Inulin and oligofructose are functional food ingredients which offer a unique combination of nutritional properties and important technological benefits. They are found in many vegetables and fruits and can be industrially obtained from chicory roots. In food formulations, inulin and oligofructose may significantly improve organoleptic characteristics. Their incorporation allows upgrading of both taste and mouthfeel in a wide range of food applications. Oligofructose is highly soluble and possesses technological properties that are closely related to those of sugar and glucose syrups. It is often used in combination with high intensity sweeteners. Inulin has a much lower solubility, improves the stability of foams and emulsions and shows exceptional fat-like characteristics when used under the form of a gel in water. Fat and carbohydrate replacement with chicory inulin and oligofructose offers the advantage of not compromising on taste and texture, while delivering nutritionally enhanced products. PMID- 12088532 TI - Oligosaccharides in infant formula. AB - Breast-feeding is the golden standard for infant feeding. However, the majority of a few week old infants are fed with a second choice infant feeding, cow's milk based formula. Amongst the multiple differences between human and cow's milk regards the development of the gastro-intestinal flora: the flora of the breast fed infant being richer in bifidobacteria and lactobacilli. Both species are known to be potentially beneficial for the health of the host. The absence of oligosaccharides, the third largest component in human milk, in cow's milk is likely to account for the differences in colonic flora. The oligosaccharide content and concentration in breast milk is - just as for the other macronutrients - a dynamic process, making it impossible for industry to mimic nature. However, if the composition cannot be mimicked, the effect and function can be imitated. The addition of two oligosaccharides, galacto-oligosaccharides and inulin, to cow's milk based infant formula has been shown to have a bifidogenic effect, and to stimulate the growth of bifidi and lactobacilli. In conclusion, the addition of oligosaccharides to cow's milk based infant formula brings this alternative, second choice infant feeding one step closer to the golden standard of human milk. But, prolonged breast-feeding should still be promoted with maximum effort. PMID- 12088533 TI - Pet food and feed applications of inulin, oligofructose and other oligosaccharides. AB - Prebiotics may be considered as functional food ingredients. They are attracting considerable interest from pet owners, pet food manufacturers, livestock producers and feed manufacturers. The most common forms of prebiotics are nondigestible oligosaccharides (NDO), including inulin, oligofructose mannanoligosaccharides, gluco-oligosaccharides, and galacto-oligosaccharides. These NDO are nondigestible by enzymes present in the mammalian small intestine, but are fermented by bacteria present in the hindgut of nonruminants. Inulin and oligofructose are present in measurable quantities in feed ingredients like wheat, wheat by-products, barley, and peanut hulls. Consumption of prebiotic oligosaccharides elicits several purported health benefits. In companion animals, prebiotics have been shown to improve gut microbial ecology and enhance stool quality. In production livestock and poultry, prebiotics are employed to control pathogenic bacteria, reduce faecal odour, and enhance growth performance. Research to date indicates positive effects of prebiotics on health status and performance of companion animals, livestock, and poultry. PMID- 12088534 TI - Communicating about gut health to the consumer: presenting the BENEO Programme. AB - Providing sufficient scientific data to be able to make 'health related claims' on foods is quite a challenge. But bringing these claims successfully to the consumer is perhaps an even bigger challenge. Especially when the claims are based on the function of the intestinal tract, as this is a subject that not everybody communicates easily about. In our consumer research, we have focused on how the new consumer thinks and talks about gut health. We found out that he is aware of the existence and the importance of gut microflora. He believes that foods can influence his own flora. At the same time, our research has tested the efficiency of several ways to communicate about these aspects. The consumer reacts quite differently on different marketing concepts. He accepts that natural ingredients can help his gut flora and expects to find such active ingredients in common everyday foods and also in diet supplements. Based on this research, we have condensed the 'prebiotic' message about inulin and oligofructose into a number of simple communications. The best of these were selected for further consumer product testing. The results of this effort form the basis of the BENEO Programme: a communication platform created by ORAFTI that allows the food industry, in partnership with ORAFTI, to bring a uniform and clear message to the consumer about the health benefits of inulin and oligofructose. PMID- 12088535 TI - Genomics and proteomics: importance for the future of nutrition research. AB - A huge number of genes within the human genome code for proteins that mediate and/or control nutritional processes. Although a large body of information on the number of genes, on chromosomal localisation, gene structure and function has been gathered, we are far from understanding the orchestrated way of how they make metabolism. Nevertheless, based on the genetic information emerging on a daily basis, we are offered fantastic new tools that allow us new insights into the molecular basis of human metabolism under normal as well as pathophysiological conditions. Recent technological advancements have made it possible to analyse simultaneously large sets of mRNA and/or proteins expressed in a biological sample or to define genetic heterogeneity that may be important for the individual response of an organism to changes in its nutritional environment. Applications of the new techniques of genome and proteome analysis are central for the development of nutritional sciences in the next decade and its integration into the rapidly developing era of functional genomics. PMID- 12088536 TI - Tunnel specificity and forager movement in subterranean termites (Isoptera: Rhinotermitidae and Termitidae). AB - The movement of foragers of two species of Australian, subterranean, mound building termites, Coptotermes lacteus (Froggatt) (Rhinotermitidae) and Nasutitermes exitiosus (Hill) (Termitidae), was investigated in their natural habitat using artificial feeding sites along trenches dug to mimic natural forager tunnels that radiate out from the central mound-nests. Termites were dyed by self-feeding on cardboard soaked with histological fat-stains on one or two trenches and then termites were collected from other feeding sites at two and four weeks after the fat-stains were placed. At two and four weeks after marking commenced, 60-75% of marked termites were found in trenches containing the marked paper, and 2-16% were found in trenches on the opposite side of the nest. The proportion of marked termites in a sample was three to eight times greater in the trenches containing the marked paper relative to other trenches. Although difficulties with fat-stains used as markers might explain some of the observed patterns, it is evident that C. lacteus and N. exitiosus foragers do not move randomly between feeding sites in their natural habitat. PMID- 12088537 TI - Insecticide use and organophosphate resistance in the coffee leaf miner Leucoptera coffeella (Lepidoptera: Lyonetiidae). AB - Increasing rates of insecticide use against the coffee leaf minerLeucoptera coffeella(Guerin-Meneville) and field reports on insecticide resistance led to an investigation of the possible occurrence of resistance of this species to some of the oldest insecticides used against it in Brazil: chlorpyrifos, disulfoton, ethion and methyl parathion. Insect populations were collected from ten sites in the state of Minas Gerais, Brazil and these populations were subjected to discriminating concentrations established from insecticide LC99s estimated for a susceptible standard population. Eight of the field-collected populations showed resistance to disulfoton, five showed resistance to ethion, four showed resistance to methyl parathion, and one showed resistance to chlorpyrifos. The frequency of resistant individuals in each population ranged from 10 to 93% for disulfoton, 53 to 75% for ethion, 23 to 76% for methyl parathion, and the frequency of resistant individuals in the chlorpyrifos resistant population was 35%. A higher frequency of individuals resistant to chlorpyrifos, disulfoton and ethion was associated with greater use of insecticides, especially other organophosphates. This finding suggests that cross-selection, mainly between organophosphates, played a major role in the evolution of insecticide resistance in Brazilian populations of L. coffeella. Results from insecticide bioassays with synergists (diethyl maleate, piperonyl butoxide and triphenyl phosphate) suggested that cytochrome P450-dependent monooxygenases may play a major role in resistance with minor involvement of esterases and glutathione S-transferases. PMID- 12088538 TI - The use of alarm pheromones to enhance bait harvest by grass-cutting ants. AB - The enhancement of bait for the control of grass-cutting ants was investigated using two species of grass-cutting ant, Atta bisphaerica (Forel) and Atta capiguara (Goncalves) (Hymenoptera: Formicidae). Bait was applied in loose piles to obtain a direct relationship between ant attraction and bait harvest. Enhancement with alarm pheromone compounds significantly increased the attractiveness and harvest of bait under certain conditions. A large proportion of the ants attracted to the enhanced bait were minor workers. These ants rarely transport bait because of their small size, and so it may be possible to increase the effect of bait enhancement by using smaller bait granules. Foragers of A. capiguara were less inclined to transport citrus-pulp bait than were those of Atta laevigata (Fr. Smith), a species that also harvests dicotyledonous plants. This emphasizes the importance of developing a bait matrix that is more acceptable to grass-cutting species. Nevertheless, the results suggest that alarm pheromone compounds have significant potential to improve the efficacy of baits for the control of grass-cutting ants. PMID- 12088539 TI - Evaluating an interpolation approach for modelling spatial variability in pest development. AB - Air temperatures estimated by partial thin plate spline interpolation, or from the 'nearest station' (Voronoi polygon method), were used to model the phenology of three pests of horticultural crops throughout England and Wales. Temperatures for a particularly hot (1976) and a particularly cold (1986) year were interpolated to a grid resolution of 1 km. Estimates were made of the timing of spring emergence (Cecidophyopsis ribis (Westwood)), the maximum number of generations completed during the summer (Plutella xylostella (Linnaeus)) and the numbers of days when mating was possible (Merodon equestris (Fabricius)). The relative accuracy of the two temperature estimation methods was compared using jack-knife cross-validation. For C. ribis and P. xylostella, modelling with interpolated temperature input data was more accurate than using data from the 'nearest station'. Of the three phenology models used, the one that relied on an activity threshold (M. equestris) was the most sensitive to both types of input data. Spatial variability in the activity of M. equestrisadults was investigated in the two main areas (south-west peninsula and Lincolnshire) where its host crop (Narcissus) is grown. Modelling at cruder scales (up to 25*25 km) masked local variation, but the degree to which this was important varied from region to region and over time, as did the geography of the variability itself. The results indicate that interpolated data, computed to a resolution of 1 km using the UK synoptic network, have the potential for wider use within agricultural decision support systems for horticultural crops. PMID- 12088540 TI - Extent of digestion affects the success of amplifying human DNA from blood meals of Anopheles gambiae (Diptera: Culicidae). AB - The success of distinguishing blood meal sources of Anopheles gambiae Giles through deoxyribonucleic acid (DNA) profiling was investigated by polymerase chain reaction (PCR) amplification at the TC-11 and VWA human short tandem repeats (STR) loci. Blood meal size and locus had no significant effect on the success of amplifying human DNA from blood meals digested for 0, 8, 16, 24 and 32 h (P = 0.85 and 0.26 respectively). However, logistic regression found a significant negative relationship between time since ingestion and the success probability of obtaining positive PCR products among meals digested for between 8 and 32 h (P = 0.001). Approximately 80% of fresh blood meals were successfully profiled. After 8 h, the proportion of blood meals that could be successfully profiled decreased slowly with time after ingestion, dropping to below 50% after approximately 15 h. There was no significant difference in the success of amplifying human DNA from blood meals of mosquitoes killed at time 0 and 8 h after ingestion (P = 0.272). PMID- 12088541 TI - Delayed mating reduces reproductive output of female European grapevine moth, Lobesia botrana (Lepidoptera: Tortricidae). AB - Virgin females of Lobesia botrana Denis & Schiffermuller were mated at ages of 1, 3, 5, 8, 12 and 16 days and the effect of mating delay on reproductive output assessed. Delayed mating did not affect female mating success but fertilization was reduced in 16-day-old females. Increased mating delays substantially affected daily oviposition pattern and resulted in a significant reduction of both fecundity and fertility, so that mean number of viable eggs laid decreased from 150 to 22 in 1- and 16-day-old mated females, respectively. Heavier females were more constrained than lighter ones by mating delays and female reproductive efficiency (no. viable eggs/female body weight) was also significantly reduced. Female longevity significantly increased and oviposition period gradually declined with mating delay. The number of viable eggs was positively correlated with both female weight and oviposition period; female longevity and female weight were also significantly correlated. However, the significance of these correlations declined with increased mating delay. Results overall indicated that mating delay drastically reduces female L. botrana reproductive output. The implications of delayed mating of females are discussed from an ecological perspective in relation to L. botrana control using mating disruption. PMID- 12088542 TI - Modelled impact of insecticide-contaminated dung on the abundance and distribution of dung fauna. AB - Deterministic models assessed the effects that contaminated dung from insecticide treated cattle had on populations of three hypothetical species of dung fauna that dispersed randomly and could double their numbers every 1-28 weeks at low density. Insecticide was allowed to kill 2-98 % of adults and prevent 16-100% of breeding in pats produced immediately after cattle treatment, with toxicity declining to < 1% in pats produced 2-23 days later. Treatment intervals were 10 40 days. The modelled impact of insecticide was affected little by approximately four-fold variations in: length and density dependence of the attractive life span of pats, frequency of pat occupation by immature adults, distribution of pat toxicity during treatment interval, and changes in dispersal rates due to age and population density. Of greater importance were variations in: pat toxicity, treatment interval, frequency of pat occupation by breeding adults, density dependence of recruitment and death, natural adversity and mortality in dormancy, general rate of dispersal, and the size and shape of the area with treated cattle. Overall, it seemed that wide variations in the impact of contamination will occur in the field, but in many situations the risk to dung fauna can be substantial, especially for slow breeding beetles, and muscoids contacting insecticide on cattle. Risk extends outside the treated areas, for a distance equal to several daily displacements of the insects. Untreated refuges for species survival should be compact blocks at least 25 daily displacements wide. PMID- 12088544 TI - Secretion clearance techniques: absence of proof or proof of absence? PMID- 12088543 TI - Field survey and greenhouse evaluation of non-rice host plants of the striped stem borer, Chilo suppressalis (Lepidoptera: Pyralidae), as refuges for resistance management of rice transformed with Bacillus thuringiensis toxin genes. PMID- 12088546 TI - Physiology of airway mucus clearance. AB - Respiratory tract secretions consist of mucus, surfactant, and periciliary fluid. The airway surface fluid is present as a bilayer, with a superficial gel or mucous layer and a layer of periciliary fluid interposed between the mucous layer and the epithelium. A thin layer of surfactant separates the mucous and periciliary fluid layers. The mucous layer extends from the intermediate airway to the upper airway and is approximately 2-10 microm thick in the trachea. Airway mucus is the secretory product of the goblet cells and the submucosal glands. It is a nonhomogeneous, adhesive, viscoelastic gel composed of water, carbohydrates, proteins, and lipids. In health, the mucous gel is primarily composed of a 3 dimensional tangled polymer network of mucous glycoproteins or mucin. Mucin macromolecules are 70-80% carbohydrate, 20% protein, and 1-2% sulfate bound to oligosaccharide side chains. The protein backbones of mucins are encoded by mucin genes (MUC genes), at least 8 of which are expressed in the respiratory tract, although MUC5AC and MUC5B are the 2 principal gel-forming mucins secreted in the airway. Mucus is transported from the lower respiratory tract into the pharynx by air flow and mucociliary clearance. Expectorated sputum is composed of lower respiratory tract secretions along with nasopharyngeal and oropharyngeal secretions, cellular debris, and microorganisms. Disruption of normal secretion or mucociliary clearance impairs pulmonary function and lung defense and increases risk of infection. When there is extensive ciliary damage and mucus hypersecretion, airflow-dependent mucus clearance such as cough becomes critically important for airway hygiene. PMID- 12088547 TI - Positioning versus postural drainage. AB - For the past 70 years positioning and postural drainage have played an important role in increasing lung volumes, perfusion, oxygenation and mobilization of secretions. While gravity is not a primary mechanism for normal secretion clearance, it plays a major role in depth and pattern of ventilation, perfusion, and lymphatic drainage. Changing patient position, or turning patients on a regular basis, is a powerful tool in maintaining lung health in a broad range of patients. In contrast, postural drainage requires considerable investment of time, and has been shown to have limited benefit in most patients. Postural drainage has been shown to improve mobilization of secretions in patients with cystic fibrosis as well as patients who produce, and have difficulty clearing, large quantities of sputum. The benefits of postural drainage appear technique dependent, requiring sufficient drainage time (3 - 15 min) for each position drained. The evidence does not support the use of vibration and percussion independent of active postural drainage. Exercise offers benefit in secretion clearance, which increases when combined with a program of postural drainage. In conclusion, routine turning, mobilization and exercise is important to maintain lung health in all patients, while postural drainage, properly applied, has been shown to improve secretion clearance in a relatively narrow range of patients with cystic fibrosis and excessive sputum production. PMID- 12088548 TI - Airway physiology, autogenic drainage, and active cycle of breathing. AB - Airway clearance techniques are used to aid in mucus clearance in a variety of disease states. Autogenic drainage and active-cycle-of-breathing technique are 2 such modalities that rely heavily on basic airway physiology to enhance clearance. In this review I discuss the equal pressure point, huffing, and asynchronous and collateral ventilation, and review the literature and theory regarding autogenic drainage and active cycle of breathing. Selection of airway clearance techniques is discussed in the light of evidence-based medicine. PMID- 12088549 TI - Positive pressure techniques for airway clearance. AB - Positive airway pressure (PAP) has been used since the 1930s to improve oxygenation, increase lung volumes and reduce venous return. More recently, PAP has been identified as an effective method of splinting airway during expiration, improving collateral ventilation, increasing response to inhaled bronchodilators, and aiding secretion clearance in patients with cystic fibrosis and chronic bronchitis. A range of devices, administration techniques, and evidence supporting their clinical use is explored, suggesting that PAP is equivalent to postural drainage in the clearance of secretions. PAP produced by threshold and fixed orifice resistors generate different characteristic flow, and airway and esophageal pressure patterns that may contribute to different physiologic effects. Further clinical studies are required to better understand the effects of these differences. PMID- 12088550 TI - High-frequency oscillation of the airway and chest wall. AB - High-frequency oscillation (HFO), applied to either the airway or chest wall, has been associated with changes in sputum attributes and clearance. The evolution of evidence, both in vitro and in vivo, supporting the use of HFO is reviewed. Devices that apply HFO to the airway range from the relatively simple mechanical Flutter and Acapella devices to the more complex Percussionaire Intrapercussive Ventilators. and the Hayek Oscillator are designed to provide high-frequency chest wall compression. Operation and use of these devices are described with examples of differentiation of device types by characterization of flows, and airway and esophageal pressures. Although HFO devices span a broad range of costs, they provide a reasonable therapeutic option to support secretion clearance for patients with cystic fibrosis. PMID- 12088551 TI - Airway clearance techniques for the patient with an artificial airway. AB - Artificial airways provide both opportunities and challenges to clinicians concerned with airway clearance. For example, the artificial airway provides direct access to the lower airways for catheter suctioning of secretions and a direct route for lung instillation of medications that promote secretion mobilization. At the same time, the presence of an artificial airway impairs natural mechanisms of airway clearance-coughing and mucociliary function. Artificial airways are invariably coated with an antibiotic-resistant bacterial biofilm that can be introduced into the lung by several commonly applied airway clearance techniques. This factor is rarely considered during research on airway clearance techniques for patients with artificial airways. This review summarizes current research on airway clearance techniques for patients with artificial airways, with special attention to the implications of the bacterial biofilm. Directions for future research are also discussed. PMID- 12088552 TI - The pharmacologic approach to airway clearance: mucoactive agents. AB - The airway mucosa responds to infection and inflammation in a variety of ways. This response often includes surface mucous (goblet) cell and submucosal gland hyperplasia and hypertrophy, with mucus hypersecretion. Products of inflammation, including neutrophil-derived deoxyribonucleic acid (DNA) and filamentous actin (F actin), effete cells, bacteria, and cell debris, all contribute to mucus purulence and, when this mucus is expectorated it is called sputum. Mucoactive medications are intended to serve one of 2 purposes; either to increase the ability to expectorate sputum or to decrease mucus hypersecretion. Mucoactive medications have been classified according to their proposed mechanisms of action. Increased knowledge of the properties of mucus has given us tools to better understand the mechanisms of airway disease and mucoactive therapy. Expectorants are thought to increase the volume or hydration of airway secretions. Systemic hydration and classic expectorants have not been demonstrated to be clinically effective. Modifiers of airway water transport are being clinically investigated as expectorants. Mucolytics degrade polymers in secretions. The classic mucolytics have free thiol groups to degrade mucin. Peptide mucolytics break pathologic filaments of neutrophil-derived DNA and actin in sputum. Nondestructive mucolysis includes mucin dispersion by means of charge shielding. Mucokinetics are medications that increase mucociliary efficiency or cough efficiency. Cough flow can be increased by bronchodilators in patients with airway hyperreactivity. Abhesives such as surfactants decrease mucus attachment to the cilia and epithelium, augmenting both cough and mucociliary clearance. Mucoregulatory agents reduce the volume of airway mucus secretion and appear to be especially effective in hypersecretory states such as bronchorrhea, diffuse panbronchiolitis, and some forms of asthma. Mucoregulatory agents include anti inflammatory agents (indomethacin, glucocorticosteroids), anticholinergic agents, and some macrolide antibiotics. Classifying mucoactive agents should help us to develop and evaluate new types of therapy and to better direct therapy toward the patients who are most likely to benefit. PMID- 12088553 TI - Airway clearance strategies for the pediatric patient. AB - Clinicians who care for cystic fibrosis (CF) patients have many techniques to choose from to facilitate mucus clearance. Little has been published about when to introduce the various techniques and in what order to teach them. Debates have occurred over these issues in the CF community, and there is now consensus on some topics. It is very important to teach adherence to therapy at an early age. Adherence to an airway clearing regimen assists in maintaining good pulmonary function in CF patients. Knowing when and how to introduce airway clearance techniques beyond chest physiotherapy (CPT) is clinically relevant and useful. A 5-position modified CPT routine can be used with infants and children, and it takes less time and may improve adherence. Infants and toddlers can be taught breathing games that eventually lead them to perform diaphragmatic breathing and huffing. Once they have mastered diaphragmatic breathing and huffing, children can be taught the active cycle of breathing technique. Modified CPT can be phased out at that point. Positive expiratory pressure therapy can usually be introduced around 6-7 years of age. High-frequency chest wall oscillation, oscillating positive expiratory pressure, and autogenic drainage all follow. CF patients should be given every opportunity to learn and master various techniques to promote mucus clearance. PMID- 12088555 TI - First interview and one session with 'ANA'. PMID- 12088556 TI - Wishing for the impossible: an essay in comparative psychoanalytic practice. PMID- 12088557 TI - Jumping from the couch: an essay on phantasy and emotional structure. AB - This paper addresses two questions: first, how do phantasies work? Second, how do these mental activities affect a person's overall emotional life? The first question tends to be overlooked since those who accept, for example, projective identification as a basic mental activity tend also to treat it as an explanatory primitive. On this view, there is no further question to ask about how projective identification itself works; rather, other psychological and emotional phenomena are explained in terms of it. By contrast, this paper asks, how does projective identification itself work? The aim is not to provide a reductive explanation but to ask how it is that phantasies have the efficacy they have. To that end, one moment in the analysis of the Rat Man is re-examined. There is then an attempt to show the difficulties involved in weaving an account of phantasy into the broader scale interpretation of emotional life. PMID- 12088558 TI - Some implications of Bion's thought: the waking dream and narrative derivatives. AB - Following up Bion's idea that dreaming is a continuous process that takes place in waking life as well as in sleep, the author develops its theoretical and technical implications. He describes the narrative derivatives of waking dream thought as the means, or interface, whereby access may be had, albeit indirectly, to the constantly forming sequences of alpha-elements. Examples of the formation of these sequences out of childhood memories, present-day experiences and sexual feelings are given. Reference is also made to the notion of the analytic field. The author goes on to demonstrate the clinical value of these concepts both for studying the session's microtransformations and as a means of in-depth examination of the analyst's own mental functioning, including its lapses, in a session. Abundant clinical material is presented to help analysts who normally apply different models of the mind share these ideas. Finally, it is suggested that night dreams result from a further elaboration of the elements formed during waking life. PMID- 12088559 TI - Pathways of pleasure: the channel of primal anality. AB - This article describes a mode of transference relationship in which the analysand has to contend with a psychic mobilisation that results from exchanges with the analyst-object and is related to the activation of pleasure in his/her own mental functioning. The patient's ego feels the internal and external excitations that stem from this mobilisation to be dangerous because of the anxieties about threatening intrusions that they raise. These anxieties arise along the contours of narcissistic flaws that give resonance to early traumatic experiences. The ego protects itself from the danger by organising narcissistic defences that oppose the impulses towards the analyst-object and foster a stagnation of the psychic work. The author puts forward the hypothesis that the patient's ego, not tolerating exposure to drive-related dangers, retracts into a mode of primary anality that is imprisoning and restrictive, setting up a fantasy of narcissistic nidification in which a part of the ego merges with an omnipotent primary object. The purpose of this strategy is to neutralise the excitations - as far as possible - while nevertheless maintaining the element of the drive excitation that forms the basis of the fantasy. The dynamics and the economy of the defensive organisation are examined in detail, as are the questions that these raise. A clinical case illustration is presented. PMID- 12088560 TI - A 9-year-old with conversion disorder, successfully treated with psychoanalysis. AB - This paper outlines the successful psychoanalytic treatment of a latency-aged boy with severe conversion disorder. Details of clinical treatment are presented. As no tested treatment has yet been found that reliably treats chronic conversion in children, this case is of importance in demonstrating the usefulness of psychoanalysis as treatment for severe psychopathology. The patient's unconscious fantasy constellations that gave rise to this syndrome are highlighted. PMID- 12088561 TI - Reporting a dream accompanying an enactment in the transference situation. AB - Patients with pathological organisations of the personality present the analyst with considerable technical difficulties. One of these problems arises from the fact that, in such patients, dreams frequently are not being used for communication of unconscious meaning, but instead for purposes of manipulation of the transference situation. They then represent attempts to identify the analyst with a part of the patient's self or with a particular internal object in order to draw him/her into collusive enactments. Following the work of Bion and Segal the paper presents a two-dimensional model in order to clarify the structure and use of dreams in this situation. The model may serve as a background orientation for the analyst in the clinical situation, as is subsequently illustrated in a detailed clinical sequence with a borderline patient. To conclude, the author suggests that whenever tendencies towards acting in are predominant, the interpretation of the enactment should generally be given preference over the interpretation of the dream content. The possible advantages and disadvantages of both strategies of interpretation are discussed. Finally, the author highlights consequences that arise when dealing with countertransference. PMID- 12088562 TI - Borges, immortality and the circular ruins. AB - The author explores ideas surrounding immortality and death focusing on the interplay between their development in two stories by Borges ('The circular ruins' and 'The immortal') and their manifestation in a patient. With the help of Borges's stories, the author addresses the desperate necessity experienced by some individuals to search for immortality. This is not just an expression of the universal wish to live forever but, at a deeper level, arises from the impossibility of bearing the mental pain of experiencing ordinary human vulnerability and loss - death being the ultimate expression of such vulnerability. It is suggested that the relentless pursuit of immortality in such individuals expresses an omnipotent phantasy of ridding the self of the emotional pain and fear that arises through being alive. It leads to a denial of the emotional significance of passage of time, of separation and sexual differences. In actuality, the individual's state of not feeling approximates to a complete loss of human identity and emotional death, with no place for any meaningful others. The individual him/herself becomes a 'mere image', living in a delusional world peopled by him/herself and his/ her projections, and ending up trapped inside the circular ruins he/she has generated. The horror experienced at the stark awareness of the individual's emotional death and the wish to re-establish contact with the good internal objects that have been attacked sets in motion the long process of searching for the recovery of a sense of temporality (that would still include the wish for immortality) and, with it, a sense of identity. PMID- 12088563 TI - The sense of beauty. AB - This paper proposes an integrative psychoanalytic model of the sense of beauty. The following definition is used: beauty is an aspect of the experience of idealisation in which an object(s), sound(s) or concept(s) is believed to possess qualities of formal perfection. The psychoanalytic literature regarding beauty is explored in depth and fundamental similarities are stressed. The author goes on to discuss the following topics: (1) beauty as sublimation: beauty reconciles the polarisation of self and world; (2) idealisation and beauty: the love of beauty is an indication of the importance of idealisation during development; (3) beauty as an interactive process: the sense of beauty is interactive and intersubjective; (4) the aesthetic and non-aesthetic emotions: specific aesthetic emotions are experienced in response to the formal design of the beautiful object; (5) surrendering to beauty: beauty provides us with an occasion for transcendence and self-renewal; (6) beauty's restorative function: the preservation or restoration of the relationship to the good object is of utmost importance; (7) the self-integrative function of beauty: the sense of beauty can also reconcile and integrate self-states of fragmentation and depletion; (8) beauty as a defence: in psychopathology, beauty can function defensively for the expression of unconscious impulses and fantasies, or as protection against self crisis; (9) beauty and mortality: the sense of beauty can alleviate anxiety regarding death and feelings of vulnerability. In closing the paper, the author offers a new understanding of Freud'semphasis on love of beauty as a defining trait of civilisation. For a people not to value beauty would mean that they cannot hope and cannot assert life over the inevitable and ubiquitous forces of entropy and death. PMID- 12088564 TI - The psychoanalytic method working with mental and physical impairment. PMID- 12088565 TI - Psychoanalysis and art: a psychoanalytic view of the life and work of Cezanne. PMID- 12088566 TI - Plenary session: 'The link and the other'. PMID- 12088567 TI - Cathecting body and mind in a new relationship: aspects of the analytic method in work with adolescents. PMID- 12088568 TI - The nature of the problems of psychoanalysis with so-called 'difficult' patients. PMID- 12088569 TI - Psychoanalysis across cultural and linguistic difference: conceptual and technical issues. PMID- 12088570 TI - Research: presentations from research advisory board funded projects. PMID- 12088571 TI - The problems of technique with severely ill patients. PMID- 12088572 TI - 'The Analyst at Work'. PMID- 12088575 TI - Contribution of nonspecific laboratory test to the diagnosis of malaria in febrile travelers returning from endemic areas: value of hypocholesterolemia. AB - BACKGROUND: The gold standard in diagnosis of malaria is microscopic detection of malaria parasites in thin blood smears. However, the sensitivity and specificity of blood smears depend mostly on the experience of the examiner. In the traveler returning from the tropics, diagnosis of malaria may be difficult when the parasitemia is low. In this circumstance any indicator that suggests the diagnosis of malaria is of great interest. The aim of this study is to determine the value of hypocholesterolemia to the diagnosis of imported malaria. METHOD: A retrospective case-control study was performed among hospitalized patients with fever returning from a malaria-endemic area, to compare the results of routine biological parameters of 129 malaria cases with those of 92 control patients. RESULTS: Multivariate analysis, using a logistic regression model demonstrates that hypocholesterolemia was the strongest parameter associated with malaria (adjusted odds ratio [OR]: 75.22, 95% confidence interval [CI] 4.60-1227.78) and the most specific (98%, 95% CI 0.95-1.0). The most sensitive abnormality was thrombocytopenia (82%, 95% CI-0.77 0.87). With a malaria prevalence of 52% in our population study, hypocholesterolemia has the strongest positive predictive value among routine biological parameters for malaria diagnosis (96%). The combination of hypocholesterolemia and thrombocytopenia was always associated with diagnosis of malaria in this study. CONCLUSION: These results show that hypocholesterolemia is significantly associated with malaria. Therefore, in the setting of negative thin and thick blood smears, the presence of hypocholesterolemia, particularly when it is combined with thrombocytopenia, in a febrile traveler returning from a malaria-endemic area, should prompt repetition and careful analysis of blood smears to avoid misdiagnosis. PMID- 12088576 TI - Combined vaccination against hepatitis A, hepatitis B, and typhoid fever: safety, reactogenicity, and immunogenicity. AB - BACKGROUND: The etiological agents of hepatitis A, hepatitis B, and typhoid fever share similar patterns of global distribution, and cause significant disease burden in travelers to endemic countries. Combined vaccination against all three diseases, based on currently available vaccines, would promote compliance and convenience for travelers. This clinical study evaluated the feasibility of extemporaneously syringe-mixed hepatitis A and B vaccine (Twinrix) and a Vi polysaccharide vaccine (Typherix) in healthy adults, and compared this to concomitant administration of the vaccines in separate arms. METHODS: The mixed dose of vaccine contained at least 720 enzyme-linked immunosorbent assay (ELISA) units of the inactivated hepatitis A antigen, 20 microg of the recombinant hepatitis B antigen and 25 microg of the Vi polysaccharide typhoid antigen in 1.5 mL. The study was conducted in 200 healthy 18- to 45-year-old volunteers. RESULTS: Equivalence between the vaccines mixed before administration and the concomitantly administered vaccines was shown in terms of seroconversion and seroprotection. With the exception of local injection site soreness, which was higher in the mixed administration group, the reactogencity was similar for both groups. In both vaccination groups more than 95% of the subjects were anti hepatitis A virus and anti-Vi seropositive 1 month after the first vaccination. With regard to hepatitis B, a strong response was achieved in both groups, with more than two-thirds of the subjects protected 2 months after the start of the immunization course. CONCLUSION: These results support the feasibility of extemporaneously syringe-mixed combined hepatitis A and B vaccine with a Vi polysaccharide typhoid vaccine, administered in healthy adults. PMID- 12088577 TI - Risk of possible exposure to rabies among tourists and foreign residents in Nepal. AB - BACKGROUND: There is little data available on the actual risk to travelers of being possibly exposed to rabies. This data would be useful in advising travelers who are considering rabies pre-exposure immunization. In addition, it is not known how many travelers are already pre-immunized when they are bitten by a possibly rabid animal. The current study was performed to determine the rate at which travelers to Nepal are possibly exposed to rabies, and to determine risk factors for possible rabies exposure. METHODS: A prospective 3-year study was carried out at the Canadian International Water and Energy Consultants (CIWEC) Clinic Travel Medicine Center in Kathmandu, Nepal, during the years 1996 through 1998. All non-Nepalese and non-Indian patients who presented with animal bites or scratches were eligible to be included in the study. RESULTS: Ninety-nine persons presented with possible rabies exposures to the CIWEC Clinic during the study period; 56 were tourists, and 43 were resident expatriates. The incidence of people presenting to the CIWEC clinic with possible rabies exposures was 1.9 per 1,000 persons/year for tourists, and 5.7 per 1,000 persons/year for resident expatriates (p <.0001). The incidence of possible exposure to rabies while trekking was 1.2 per 1,000 persons/year. Women were significantly more likely than men to present with a possible rabies exposure, accounting for 61% of patients (p =.0027). Younger people were more likely to have bite exposures to the face and head than older patients. The length of time between exposure and treatment averaged 1.6 to 5.0 days. Among patients presenting with animal bites, 56% of foreign residents, and 21% of tourists had been pre-immunized against rabies. CONCLUSIONS: Foreign residents of Nepal are significantly more likely to be exposed to rabies than tourists. Trekking does not increase the chances of being exposed to rabies. Children have a higher risk of being bitten on the face and head, and females are more likely than males to be bitten or scratched by a possibly rabid animal. PMID- 12088578 TI - Travel health: a survey of life jacket designs currently in use on commercial aircraft. AB - BACKGROUND: Although any travel harbors potential health problems, travel across water poses the additional risk of involuntary contact with water in the event of an emergency. Studies suggest that fatal boat-related accidents have occurred due to the passengers' inability to don their life jackets. With the dramatic increase of long-haul flights, it was decided to investigate if potential safety hazards can be identified leading to similar problems in relation to the life jackets provided onboard aircraft. It was assumed that the variety of designs combined with a possible lack of attention paid to safety instructions could represent such a problem. METHOD: Safety instruction cards in passengers' seat pockets usually depict the life jacket model used on a particular aircraft. Ninety-eight such safety cards from 53 airlines were analyzed and categorized to identify the range and variety of designs currently in use. RESULTS: Twelve different categories of life jacket models could be distinguished with some airlines using more than one model. It was also found that not all cards show a clear depiction of the models used, the cards of two airlines show two different designs on the same cards, and two other companies chose to change the model halfway through the pictured donning procedure. CONCLUSION: There is a wide variety of life jacket designs in use with the safety cards not always giving accurate instructions. This variety also has implications for the safety demonstration at the beginning of a flight. Further research is recommended into a range of topics, one being the quality and content of these instructions, and also into other personal floating devices provided for the safety of travelers in the event of an emergency over water. PMID- 12088579 TI - Enterotoxigenic Escherichia coli as cause of diarrhea among Mexican adults and US travelers in Mexico. AB - BACKGROUND: Enterotoxigenic Escherichia coli (ETEC) is the most common pathogen identified in travelers to Mexico with diarrhea. There have been few recent studies looking at the etiology of diarrhea in travelers compared with the local resident population. METHODS: We compared enteric pathogens isolated in two populations experiencing acute diarrhea acquired in Guadalajara, Mexico and also compared clinical illness caused by the principal pathogen, ETEC. RESULTS: A single and 2 enteropathogens were detected in 107 (23%) and 8 (2%), respectively, of 457 Mexicans in 1995 and 1997, and 37 (29%) and 2 (2%), respectively, of 127 US adults in 1997. The most common pathogen was ETEC in both groups (11% of Mexican, 19% of US adults), although more common in the US travelers group (p =.0017). Shigella spp and Cryptosporidium spp were less common in the Mexican (<1% and <1%, respectively) than in the travelers group (6% and 3%, respectively) (p <.001 and p =.002, respectively). Entamoeba histolytica was more often found in the Mexican group (4% Mexican, 0% US adults; p =.027). CONCLUSION: ETEC is the most common pathogen among travelers and Mexican residents in this study. The duration of untreated diarrhea due to ETEC was significantly shorter among Mexicans (49 hours in Mexican, 94 hours in US adults; p =.0004), as was the average number of unformed stools passed over 4 days (Mexicans 8.8 versus travelers 17.9 stools; p =.0009 PMID- 12088580 TI - Traveling with infants and young children. Part III: travelers' diarrhea. PMID- 12088581 TI - Diarrhea outbreak among Czech Army Field Hospital personnel caused by Providencia alcalifaciens. PMID- 12088582 TI - Four-site intradermal postexposure boosters in previously rabies vaccinated subjects. PMID- 12088583 TI - Trends in antimalarial drugs prescribed in New Zealand 1993 to 1998. PMID- 12088584 TI - Knowledge, attitudes, and practices on malaria preventive measures of migrants attending a public health clinic in northern Italy. PMID- 12088585 TI - A travel medicine practitioner's tale. PMID- 12088586 TI - Spotting the spirochete: rapid diagnosis of leptospirosis in two returned travelers. PMID- 12088587 TI - Vaccination against meningococcal disease: which vaccine to use? PMID- 12088589 TI - Simple polynomial multiplication algorithms for exact conditional tests of linearity in a logistic model. AB - The linear logistic model is often employed in the analysis of binary response data. The well-known asymptotic chi-square and likelihood ratio tests are usually used to detect the assumption of linearity in such a model. For small, sparse, or skewed data, the asymptotic theory is however dubious and exact conditional chi square and likelihood ratio tests may provide reliable alternatives. In this article, we propose efficient polynomial multiplication algorithms to compute exact significance levels as well as exact powers of these tests. Two options, namely the cell- and stage-wise approaches, in implementing these algorithms will be discussed. When sample sizes are large, we propose an efficient Monte Carlo method for estimating the exact significance levels and exact powers. Real data are used to demonstrate the performance with an application of the proposed algorithms. PMID- 12088588 TI - Objective assessment of blood stasis using computerized inspection of sublingual veins. AB - In traditional Chinese tongue diagnosis, inspection of the sublingual veins has been developed as a new diagnostic examination method especially for determining blood stasis. The main purpose of this research is to develop a computerized inspection system, used to quantitatively extract the chromatic and geometrical properties of sublingual veins. A method based on color equalization is presented to assist the image segmentation of sublingual veins. Color image analysis techniques are applied to extract the length, width, area and color information from sublingual veins. Experimental results are compared with the visual inspection of physicians. The severity of blood stasis was classified into three groups: normal, moderate and pronounced. Each group consisted of eight subjects. The overall recognition rate is 87.5% in the leaving-one-out classification using the developed system. PMID- 12088590 TI - The use of simulated annealing for finding optimal population designs. AB - The development of functions for MATLAB and S-PLUS that can be used for the evaluation of specific population pharmacokinetic designs has been described recently. These functions are based on the evaluation of an approximation of the population Fisher information matrix. Optimisation of the design of the population experiment can be made on the basis of D-optimal design techniques, where the determinant of the population Fisher information matrix is maximised. This maximisation is complex due to the convoluted nature of the surface of the determinant. Four optimisation algorithms (simplex, non-adaptive random search, non-adaptive random search followed by simplex and simulated annealing) are compared in their ability to optimise the sampling times for various design structures for three examples of population pharmacokinetic models. In all cases, despite more computing time, simulated annealing was superior to the other methods for finding optimal designs with greater benefits being seen over the other algorithms for the more complex designs. PMID- 12088591 TI - Rook methodology: a flexible framework for exact test development in the matching paradigm. AB - As well as supplying a means of enumerating permutations, rook methodology provides a flexible and fertile analytic framework that can facilitate statistical test development in applied settings. Four applications in psychology, where rook methodology provides exact small-sample tests, are described. They are (a) matching under uncertainty, (b) ordered spatial memory, (c) rater agreement under uncertainty, and (d) multiple object relocation. An online computer program, that supplies an exact small-sample test of hypotheses within the rook paradigm, is outlined. The program, which exploits rook methodology to enumerate permutations, may be accessed and executed via the Internet using an ordinary browser. PMID- 12088592 TI - System-approach methods for modeling and testing similarity of in vitro dissolutions of drug dosage formulations. AB - System-approach based modeling methods are used to model dynamic systems describing in vitro dissolutions of drug dosage formulations. Employing the models of these systems, model-dependent criteria are proposed for testing similarity between in vitro dissolutions of different drug dosage formulations. The criteria proposed are exemplified and compared with the criterion called the similarity factor f(2), commonly used in the field of biomedicine. Advantages of the criteria proposed over this factor are presented. PMID- 12088593 TI - Development of a digital signal processor-based new 12-lead synchronization electrocardiogram automatic analysis system. AB - This paper presents a digital signal processor (DSP)-based new multichannel electrocardiogram (ECG) system for 12-lead synchronization ECG automatic analysis in real-time with high sampling rate at 1000 Hz and 12-bits precision. Using the hardware structure of double-CPU based on Microprocessor (MPU) 89C55 and DSP TMS320F206 combines the powerful control ability of MPU with DSPs fast computation ability. Fully utilizing the double-CPUs resource, the system can distribute the reasonable CPU-time for the real-time tasks of multichannel synchronization ECG sampling, digital filter, data storing, waveform automatic analysis and print at high sampling rate. The digital ECG system has the advantages of simple structure, sampling with high speed and precision, powerful real-time processing ability and good quality. The paper discusses the system's principle and the skilful hardware design, also gives the ECG processing using the fast simple integer-coefficient filter method and the automatic calculation algorithms of the ECG parameters such as heart rate, P-R interval, Q-T interval and deflexion angle of ECG-axis etc. The system had been successfully tested and used in the ECG automatic analysis instrument. PMID- 12088594 TI - Parametric receiver operating characteristic curve analysis using mathematica. AB - Several computer programs have been written to perform receiver operating characteristic (ROC) curve analysis, and are available in the public domain. Here, the author provides the theory and description for 'rocMath', a Mathematica program that performs parametric ROC curve analysis. The 'rocMath' program has some advantages over other ROC curve programs, including the ability to provide, through optional arguments: (a) user-specified pointwise confidence limits, as well as default 95% limits, on ROC curve area and on true-positive rates; (b) ROC curve plots with data points, a fitted curve, and user-specified pointwise confidence bands; and (c) ROC curve areas, tables, and plots based on a logistic distribution as well as on a standard normal distribution. In addition, the code of 'rocMath' can be modified to address additional ROC curve applications. The program uses Mathematica's ability to operate on purely symbolic as well as numeric data to achieve substantial coding efficiency. Limitations of the 'rocMath' program are also discussed. PMID- 12088595 TI - A web-based self-monitoring system for people living with HIV/AIDS. AB - Available evidence indicates that the internet is becoming the main source for delivering information on the prevention, control and treatment of HIV/AIDS. This paper presents a further step in the use of the internet to meet the new challenge of managing HIV as a chronic illness. It describes a work carried out under the SEAHORSE EU project to build and test a Web-based self-monitoring system for HIV/AIDS patient care. The user interface has been carefully designed to provide a high-level of interaction and therefore improve some of the current limitations of Web applications. The system comprises three modules: (1) a patient self-monitoring personal diary, to create a follow-up patient record; (2) a data analysis and visualisation tool; and (3) a section to allow patients to ask for advising and remote doctor support. One of the crucial system design issues has been the system security and users anonymity features required in this clinical domain. A feasibility pilot has been carried out to test the system in three organisations involved in the research project: the Immune Development Trust (London), the Lambeth, Southwark and Lewisham Health Authority (London) and Apoyo Positivo in Madrid. The preliminary results of the system evaluation show the potential validity and usefulness of the tool for helping people living with HIV/AIDS to promote and manage their health and for providing health professionals with new means for tele-monitoring and tele-caring patients. PMID- 12088596 TI - Research on patient needs in palliative care at the end of life. PMID- 12088597 TI - Problems to discuss with cancer patients in palliative care: a comprehensive approach. AB - This study presents a comprehensive overview of the problems patients experience in a palliative phase of cancer. A two-step qualitative method was used: in-depth interviews with patients and relatives (N=9 patients+7 relatives), followed by interviews (N=31patients+15 relatives) using a checklist to confirm and complete the picture. Quality of life and quality of care domains were addressed. Patients experienced problems in all of these palliative care domains, although individual patients may have experienced only a few problems. Fatigue, feelings of futility, reluctance to accept help, fear of suffering and the fear that help would not be available if needed, were common problems. Communication problems arose when a grudge against a GP had remained, or because one family member tried to spare the other a confrontation with his/her feelings of fear or grief. A comprehensive checklist of relevant problems is presented. PMID- 12088598 TI - Cancer and autonomy. AB - The idea to be reflected upon in this article concerns choice as a basic act in case of illness. Choices are precondition for decision making, decision making is crucial during the course of an illness. The making of choices depends on autonomy as a characteristic of identity. This article describes different psychological characteristics of autonomous behavior, some options for the health professional to accordingly interact with the patient, his history and future and the consequences for the provoking of stress during the diagnostic and treatment procedures. Stress remaining an important and probably decisive factor in the course of an illness like cancer. PMID- 12088599 TI - Communication difficulties of healthcare professionals and family caregivers of HIV infected patients. AB - The family caregivers of HIV infected patients play a growing role in the support of the everyday life of their patient. Being more and more involved in different aspects of counselling and care-giving, they become the partner of the healthcare professionals who take care of the patient. It is nowadays well known that communication between healthcare professionals and patients is complex. So communication is between healthcare professionals and family caregivers, as our survey aims at demonstrating. Both the family caregivers and the healthcare professionals face difficulties and express needs and expectations that are important to meet in order to optimise the quality of care which is given to the patient. PMID- 12088600 TI - A questionnaire study of GPs' and patients' beliefs about the different components of patient centredness. AB - Much research has explored the interaction between doctor and patient in the consultation and patient centredness has generally emerged as the preferred mode of consultation style. The present study aimed to examine and compare general practitioners' (GPs) and patients' beliefs about the importance of the different aspects of patient centred behaviour in a consultation. Matched questionnaires were completed by 410 patients (response rate=76.5%) and 64 GPs (response rate=85.3%) from practices around London concerning aspects of patient centredness operationalized in terms of doctor receptiveness, patient involvement, the affective content of the relationship and information giving. The results showed that GPs and patients showed similar beliefs about involving the patient in decision making, aspects of doctor receptiveness and the importance of the patient's own feelings in the consultation. However, GPs believed that it was less important to focus only on the patient's main problem, and more important to acknowledge their own feelings and avoid medical language. Further, GPs rated doctor receptiveness and the affective content of the relationship overall as more important for a good consultation than the patients. The patients also consistently rated information giving as more important than the GPs. To conclude, GPs rated the doctor receptiveness and affective components of patient centredness as more important than patients whereas patients showed greater endorsement of information indicating that although patient centredness may currently be the preferred style of consultation, doctors and patients prefer different aspects of this behaviour. PMID- 12088601 TI - Development and feasibility of a modified reattribution model for somatising patients, applied by their own general practitioners. AB - Reattribution has been developed as a cognitive-behavioural treatment model for somatisation in general practice. Our objective is to make reattribution suitable for application on patients with long-standing somatisation, including hypochondria, and to evaluate feasibility. Three modifications were developed: (1) dealing with persistent illness worry, (2) adjustment of the doctor's speed to that of the patient, and (3) the use of symptom diaries. Performance of ten experienced general practitioners (GPs), after a 20h training programme (six sessions of variable length), was measured by self-registrations and audio-taped consultations. GPs were interviewed on factors interfering with performance. Nine GPs completed the course. Reattribution was applied to 51 out of 75 indicated somatising patients, which required on average three consultations of 10-30min duration. We conclude that the modified reattribution model offers a feasible approach to the broad spectrum of somatisation seen in general practice; only the modification 'dealing with illness worry' showed limited feasibility. PMID- 12088602 TI - 'Health repertories': an understanding of lay management of minor ailments. AB - Data from a programme of research exploring information channels about health and medicines used by young adults (16-24 years old) in the UK were considered, in the light of existing schema and memory theory, to provide a concept of 'health repertories' for self-medication of minor ailments. Focus groups comprising a total of 48 young adults were conducted, followed by structured interviews with 76 young adults who visited a community pharmacy to purchase non-prescription medicines or to ask for advice, and case studies from this cohort. We propose that young adults develop a dynamic 'health repertory' of information for management of minor illness episodes, comprising a number of 'entries', and that these repertories are consistent with schema theory. Each 'repertory' includes description/labelling of symptoms, one or more self-medication strategies, and contingency plans (including formal health care intervention), if these strategies fail. Information in the repertory is drawn from both lay and professional channels. PMID- 12088603 TI - Development and pilot testing of a decision aid for postmenopausal women with osteoporosis. AB - This study's aim was to develop and pilot test an evidence-based decision aid for postmenopausal women with osteoporosis who are considering options to prevent fractures. The aid was based on the Ottawa Decision Support Framework, and integrated evidence from our Cochrane systematic reviews. Following development by a panel of experts in osteoporosis and decision making, a user review panel of practitioners and women who had already made their decision about osteoporosis therapy reviewed the decision aid for acceptability. Then the decision aid was pilot tested using a before-after design in women at the point of decision making. Compared to baseline, there were statistically significant improvements in knowledge, realistic expectations and decreased decisional conflict. Our decision aid shows promise in preparing women for counseling about osteoporosis therapies. Long-term adherence to chosen therapy and quality of life will be evaluated in a randomized controlled trial. PMID- 12088604 TI - Improving patient involvement in chronic disease management: the views of patients, GPs and specialists on a guidebook for ulcerative colitis. AB - Patient information, shared care and decision-making are recognised as beneficial to chronic disease management. As part of an RCT, opinions of ulcerative colitis patients and their doctors were sought on a guidebook designed to support self care and mediate doctor/patient interaction during consultations. Semi-structured interviews were conducted with 6 specialists and 16 GPs. Patients' views were obtained from written responses to a postal questionnaire (136 replies). Inclusion of lay knowledge and clarification of treatment decision processes increased patients' perception of involvement in disease management through increased identification and awareness of others' self-management experiences. Doctors focused on the importance of improving patients' compliance and use of services. However, the guidebook was seldom used as it had been intended during consultations. Patients felt constrained by time limits and consultants did not actively encourage guidebook use. Based on the findings, we suggest an approach utilising the guidebook to improve patient participation in disease management. PMID- 12088605 TI - Factors related to fear in patients undergoing cataract surgery: a qualitative study focusing on factors associated with fear and reassurance among patients who need to undergo cataract surgery. AB - The aim was to identify factors that are related to fear among patients who need to undergo cataract surgery. The data were collected by focus group interviews (n=27). The doctor-patient relationship, patient education, the wait, hospital organization, social support, sensations, previous experience, outcome of surgery, and coping strategies were identified as the main factors that contribute to feelings of fear related to cataract surgery. Five stages of fear were identified: at home after diagnosis, during preparation for surgery at the hospital, the day of surgery, the post-operative visits, and the period after these follow-up visits at home. A model regarding the factors related to fear in patients awaiting cataract surgery was developed, which emphasizes the importance of a good doctor-patient relationship, and the need for patient education that is tailored to the individual patients. PMID- 12088606 TI - Evaluating the reliability of DISCERN: a tool for assessing the quality of written patient information on treatment choices. AB - Patients require good quality, evidence based information so that they can participate actively in the decision making process. The DISCERN instrument has been developed to help patients rate the quality of written information materials about treatment choices. This study evaluated the reliability of DISCERN using 31 information leaflets discussing treatment options for prostate cancer. The index of agreement between two independent raters was substantial for the overall quality rating (kappa=0.65, 95% confidence interval 0.49, 0.82), indicating that the instrument could be used to discriminate reliably between low and high quality prostate cancer publications. Healthcare professionals should inform patients of the availability of the instrument and encourage its use by patients who regularly attend to written sources of information on treatment choices. Early indications show that DISCERN could enable both patients and healthcare professionals to discriminate between the plethora of variable quality information currently available. PMID- 12088608 TI - Antioxidant enzymes and lipid peroxidation in the scalp of patients with alopecia areata. AB - Alopecia areata (AA) is an autoimmune inflammatory disease. However, little is known about the alterations in lipid peroxidation and antioxidant enzymes in the scalp of patients with AA. Therefore, the aim of this study was to investigate the status of oxidative stress in the scalp of patients with AA. We measured the levels of thiobarbituric acid reactive substances (TBARS) as lipid peroxidation status, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) as antioxidant enzymes in the scalp of ten patients with AA and ten control subjects. The levels of TBARS in scalp of patients with AA (3654.1+/-621.2 nmol/g tissue) were significantly higher than those of controls (1210.2+/-188.8 nmol/g tissue) (P=0.002). The levels of SOD (134.8+/-23.8 U/g tissue) and GSH-Px (332.7+/-66.2 U/g tissue) in scalp of patients with AA were also significantly higher than those of controls (63.2+/-8.8 U/g tissue, 112.0+/-18.4 U/g tissue, respectively) (P=0.019, P=0.002, respectively). The mean levels of TBARS, SOD and GSH-Px in early phase of disease were increased 2-fold as compared with late phase of the disease. These results indicate that oxidative status is affected in AA. Lipid peroxidation and antioxidant enzymes may be involved in the pathogenesis of AA. Furthermore, we found high SOD and GSH-Px activities in the scalp of patient with AA. These high levels could not protect the patients against the reactive oxygen species, because lipid peroxidation could not be lowered in AA patients. PMID- 12088607 TI - Cutaneous late phase reaction in adult atopic dermatitis patients with high serum IgE antibody to Dermatophagoides farinae: correlation with IL-5 production by allergen-stimulated peripheral blood mononuclear cells. AB - It is known that, in patients of allergic asthma and rhinitis, the late-phase reaction (LPR) occurs 6-12 h after allergen challenge, but there are few reports concerning cytokine production in the cutaneous LPR in atopic dermatitis (AD). We report here the results of our study on the relationship between the cutaneous LPR and the production of cytokines such as IL-4, IL-5, IL-2 and IFN-gamma by peripheral blood mononuclear cells (PBMC) of AD patients. We selected 29 pure AD patients with no history of atopic airway diseases who showed high serum IgE antibody against Dermatophagoides farinae and performed skin prick testing with three different antigens and observed the resultant cutaneous reactions in 23 of the AD patients. Furthermore, we measured the cytokine production by the cultured PBMC under the stimulation of the antigens and compared it with the results of the skin tests. 13 (57%) of these 23 AD patients demonstrated positive LPR in response to D. farinae, and the mean concentration of IL-5 produced by PBMC was higher in these LPR-positive AD patients compared to the LPR-negative ones. Additionally, we noticed that there was a positive correlation between the mean diameter of the erythema of LPR and the level of IL-5 production by PBMC in the LPR-positive patients. We suggest that there are at least two groups in AD patients, i.e. LPR-positive and LPR-negative ones. The observation of LPR can be an important and practical way to classify AD patients into subgroups, which may enable us to regard IL-5 or eosinophils as a target for treatment. PMID- 12088609 TI - Videomicroscopic and histopathological investigation of intense pulsed light therapy for solar lentigines. AB - A noncoherent, broadband, intense pulsed light source has been effective for symptoms of photoaging skin as a nonablative method. The purpose of this study was to investigate the mechanism of efficacy of intense pulsed light for solar lentigines, a symptom of photoaging skin, with videomicroscopy and histopathology. Skin lesions of patients with solar lentigines who received one treatment of intense pulsed light were examined. Sixteen of 20 patients showed tiny crusts clinically. These tiny crusts were confirmed to be micro-crust formation after epidermal injury with sequential observation using videomicroscope and histopathology. Drop-off of micro-crusts with ample melanin pigments lead to clinical improvement of skin lesions. Intense pulsed light with absorption spectrum for melanin induced injury of melanin-containing epidermal cells via photothermal effects, suggesting that intense pulsed light may be a new modality for solar lentigines. PMID- 12088610 TI - Propionibacterium acnes-induced IL-8 production may be mediated by NF-kappaB activation in human monocytes. AB - Propionibacterium acnes (P. acnes) causes an inflammatory acne that is characterized by massive neutrophilic infiltration. IL-8 is thought to play an important role in the pathophysiology of P. acnes, although the mechanisms by which P. acnes up-regulates the release of IL-8, a neutrophilic chemokine, from target cells is not well understood. In this study, we investigated the mechanisms through which heat-killed P. acnes induces IL-8 production in THP-1 cells (a human monocytic cell line). We found that P. acnes is able to directly induce IL-8 production and IL-8 mRNA expression in human monocytic cells in a dose- and time-dependent manner through a mechanism requiring transcription factor NF-kappaB activation. Additionally, P. acnes-induced IL-8 secretion was inhibited by roxithromycin, a macrolide antibiotic, and its inhibitory effect seemed to be partially associated with the inhibition of P. acnes-induced NF kappaB activation. This is the first study to show that NF-kappaB activation is involved in the IL-8 production of monocytic cells stimulated by P. acnes. PMID- 12088611 TI - The effects of basic fibroblast growth factor (bFGF) on the breaking strength of acute incisional wounds. AB - Many researchers have reported the effects of cytokines on ulcers, especially on chronic ulcers. However, the long-term influences of cytokines on acute incisional wounds with respect to breaking strength, quality of the scar etc. have not been elucidated completely. In the present study, the breaking strength of scars produced by full thickness incisional wounds of the skin created on backs of rabbits were compared when the wounds were treated by conventional suturing or with basic fibroblast growth factor (bFGF) in addition to suturing. We administered bFGF to full thickness acute incisional wounds of the skin created on backs of rabbits. The breaking strengths of wounds treated by conventional suturing and those treated with bFGF soon after the operation were compared as were the conditions of the scars after long-term follow up of up to 7 weeks. They were also examined histopathologically. Administration of bFGF at the time of wound closure, especially 1.0 microg in amount per cm, increased the breaking strength significantly compared to the control group from 5 weeks after the operation. Moreover, histopathological examination revealed that there was rich vascularization soon after the operation and an arrangement of collagen fibers which lenticulated horizontally from the 5th week and later after the operation in the bFGF treated groups in a dose dependent manner. This study demonstrates that administration of bFGF at the time of wound closure significantly increased the breaking strength compared to the control group from 5 weeks after the operation. And the administration of bFGF also improved the quality of the scar, the condition of the scar by surface condition by inspection as well as the width of scar at histological evaluations. So we think this cytokine may be highly effective clinically. This may be especially true for patients whose wound healing process tends to be delayed such as aged patients or when high breaking strength is recommended even for patients with normal wound healing potential. PMID- 12088614 TI - Spectrum of vibrio species associated with acute diarrhea in North Jakarta, Indonesia. AB - Vibrio spp was isolated from 1024 (21.2%) of 4820 diarrhea patients admitted to a community hospital in North Jakarta from 1996 through 1998. Vibrio cholerae O1 (49.5%) and V. parahaemolyticus (30.1%) comprised the major species isolated, followed by V. cholerae non-O1 (16.9%), and V. fluvialis (9.4%). In 938 (19.4%) patients, Vibrio was found as single isolate. Multiple infections were detected in 86 (1.8%) patients. A small number of V. furnisii, V. metschnikovii, V. mimicus and V. hollisae were also isolated. No V. cholerae O139 was detected. The majority of patients with Vibrio spp. infections were adults between the ages of 20 to 45 years. No Vibrio spp. was isolated from infants <1 year old in this study. In vitro antibiotic susceptibility testing revealed no antibiotic resistance associated with the 507 V. cholerae O1 isolates tested, except for colistin (100%). These data implicate Vibrio spp. as a major cause of diarrhea in this region. PMID- 12088615 TI - Culturing bronchial washings obtained during bronchoscopy fails to add diagnostic utility to culturing the bronchoalveolar lavage fluid alone. AB - A retrospective analysis was performed to determine whether cultures of bronchial washings (BW) obtained during bronchoscopy added to the diagnostic efficiency of cultures of bronchoalveolar lavage fluid (BAL) alone. Results of BW and BAL cultures submitted from 268 patients over a 7-month period were compared. The isolation of an organism from the BW but not from the BAL occurred in only 17.4% of cases. Moreover, the vast majority of those organisms consisted of yeasts or molds of questionable clinical significance that did not prompt a change in antimicrobial therapy. Culturing the BAL specimen alone would have resulted in an efficiency of 97.0% (95% confidence interval 94.2-98.7%) for the isolation of clinically relevant pathogens identified from bronchoscopic specimens. These results suggest that the submission of BW obtained during the BAL procedure for culture evaluation not only fails to add diagnostic value, but may also result in unnecessary laboratory evaluations and provide misleading information to clinicians. PMID- 12088616 TI - Comparison of NCCLS microdilution method and Etest in antifungal susceptibility testing of clinical Trichosporon asahii isolates. AB - We investigated the in vitro activity of amphotericin B, fluconazole, and itraconazole against clinical Trichosporon asahii isolates (n = 43) by NCCLS M27A reference microdilution method and explored the correlation between Etest and NCCLS reference method. Microdilution MIC ranges following 48 h of incubation were 1-8, 0.25-16, and 0.06-4 microg/ml for amphotericin B, fluconazole, and itraconazole, respectively. The corresponding Etest MIC ranges were determined as 0.125- > 8, 0.25- > 64, and 0.03-8 microg/ml. Of interest, Etest tended to produce lower amphotericin B MICs and widen the MIC range compared to microdilution. The influence of Etest on fluconazole and itraconazole MICs was in contrary with that observed for amphotericin B. Etest MICs of fluconazole and itraconazole tended to be higher than microdilution MICs. The wider range of amphotericin B MICs obtained by using Etest methodology may facilitate discrimination of isolates with reduced susceptibility to amphotericin B. However, clinical significance of these findings remain yet unknown and determination of MIC breakpoint values is required. PMID- 12088617 TI - Diagnosis of pelvic actinomycosis by 16S ribosomal RNA gene sequencing and its clinical significance. AB - Traditional ways of identification of anaerobic Gram-positive non-sporulating bacilli by isolation of the organism and studying it phenotypically by elucidation of its morphologic and biochemical characteristics and metabolic end products are associated with a need for special equipment and expertise, and strains that are "unidentified" because of ambiguous biochemical profiles. In this study, an anaerobic Gram-positive non-sporulating bacterium was isolated from the intrauterine contraceptive device of a 36-year old woman with pyosalpinx. The Vitek system (ANI) showed that it was 99% Propionibacterium granulosum; whereas the API system (20A) showed that it was 78% Actinomyces meyeri/odontolyticus. The 16S ribosomal RNA gene of the strain was amplified and sequenced. There was 0 base difference between the isolate and A. odontolyticus (GenBank Accession no. AJ234047), indicating the isolate most closely resembled a strain of A. odontolyticus. Identification of the organism in this study was important because the duration of antibiotic therapy would be entirely different. In the present case, identification of the bacterium as A. odontolyticus inferred that the patient suffered from an intermediate form of pelvic actinomycosis. A prolonged course of antibiotics would be more desirable, as the relapse rate of actinomycosis after a short course of antibiotics is high. PMID- 12088618 TI - Value of Etest penicillin V and penicillin G strips for penicillin susceptibility testing of Neisseria meningitidis. AB - The NCCLS agar dilution method and Etest are currently accepted methods for susceptibility testing of Neisseria meningitidis to penicillin. We determined the MIC of penicillin V and penicillin G by both the agar dilution method and Etest using 43 strains of N. meningitidis. Although results for reference strains were within the accepted quality control range of penicillin MICs for both drugs, differences of two to three dilutions were seen between the two antibiotics with both methods. Penicillin V results cannot correctly predict the susceptibility to penicillin G for N. meningitidis if penicillin G breakpoints are used for penicillin V. However, adjusting the penicillin V breakpoints two dilutions higher (i.e., S < or = 0.25 and R > or = 8 microg/ml), concordance could be achieved for susceptibility categorization by the two compounds. An agreement of 98% +/- 1 dilution was obtained between Etest and the reference method when using penicillin G strips. We conclude that Etest with penicillin G strips is a convenient and reliable alternative method for determining the MICs of penicillin for N. meningitidis. PMID- 12088619 TI - Mupirocin resistance in methicillin-resistant Staphylococcus aureus clinical isolates in a Spanish hospital. Co-application of multiplex PCR assay and conventional microbiology methods. AB - A total of 1785 Staphylococcus aureus clinical isolates were collected in our hospital during 1998 (526), 1999 (564) and 2000 (695). Among them, one hundred and thirty (39, 33 and 58, respectively) were phenotypically assigned as methicillin-resistant Staphylococcus aureus (MRSA); sixteen of these isolates (3, 2 and 11, respectively) were detected as highly mupirocin-methicillin resistant Staphylococcus aureus (MMRSA). In this work, our goal was to characterize MRSA and MMRSA clinical isolates by co-application of phenotypic and genotypic methods in order to determine the MMRSA incidence in our hospital during the period 1998 2000. With this purpose we compared and integrated the results obtained using conventional microbiology methods with those obtained using a multiplex polymerase chain reaction (PCR) assay. Our results showed a good complementation between these two approximations to determine the incidence of MMRSA clinical isolates and permitted to estimate that such incidence increased from 7.7% in 1998 to 19% in 2000. PMID- 12088620 TI - Comparative in vitro activities of ertapenem against aerobic and facultative bacterial pathogens from patients with complicated skin and skin structure infections. AB - This study compared the in vitro activities of ertapenem (Merck & Co., Inc.), ceftriaxone, amoxicillin-clavulanate, and piperacillin-tazobactam against 518 aerobic and facultative bacterial pathogens isolated from 340 patients with complicated skin and skin structure infections. Ciprofloxacin was also tested against Gram-negative isolates. Gram-positive cocci accounted for 68.1% of the aerobic bacteria; Staphylococcus aureus was the most common isolate (45.6%). The ertapenem MIC was < or = 2 microg/ml for 80.9% of isolates and > or = 8 microg/ml for 16.2% (including isolates of enterococci, methicillin-resistant S. aureus, Pseudomonas aeruginosa, and other nonfermentative Gram-negative bacteria). Against methicillin-susceptible S. aureus, ertapenem had the most potent activity. Ertapenem was the most active drug against Enterobacteriaceae (100% susceptible), whereas amoxicillin-clavulanate was least active (66% susceptible). Piperacillin-tazobactam was the most active drug against P. aeruginosa (100% susceptible), followed by ciprofloxacin (87% susceptible). In summary, ertapenem was highly active in vitro against many aerobic and facultative bacterial pathogens commonly recovered from patients with complicated skin and skin structure infections. PMID- 12088621 TI - Validation of Vitek version 7.01 software for testing staphylococci against vancomycin. AB - We tested 143 isolates of staphylococci with vancomycin by the National Committee for Clinical Laboratory Standards broth microdilution (BMD) reference method and compared the results to those generated using the Vitek automated system (GPS-105 and GPS-107 cards and version 7.01 software). For ten isolates, the vancomycin MICs by BMD were 8 microg/ml. By Vitek, the vancomycin MICs ranged from 2 to 16 microg/ml. Vancomycin MICs of > or =32 microg/ml were reported for two additional isolates by Vitek; however, the MICs decreased to < or =0.5 microg/ml on retesting. By BMD, the vancomycin MICs for both isolates were 1 microg/ml. While the modal vancomycin MIC results by BMD for S. aureus and coagulase-negative staphylococci (CoNS) were both 1 microg/ml, Vitek results showed a mode of < or =0.5 microg/ml for S. aureus, and a mode of 2 microg/ml for CoNS. Vitek did not report vancomycin MICs of 1 or 4 microg/ml for any of the isolates tested. While the sensitivity of detecting staphylococci with reduced susceptibility to vancomycin appears to be improved with Vitek version 7.01 software, when compared to earlier software versions, laboratories may notice an overall shift in MIC data toward higher vancomycin MICs, although for the most part, this does not affect the categorical interpretations of the results. PMID- 12088622 TI - Emerging fluoroquinolone-resistance for common clinically important gram-negative bacteria in Taiwan. AB - The in vitro antimicrobial activity of the three most commonly used fluoroquinolones (norfloxacin, ofloxacin and ciprofloxacin) was measured for 2235 isolates of seven common pathogenic gram-negative bacteria. Collected from a major teaching hospital in Taiwan during the years 1985-86, 1989-90, and 1996-97, the samples were evaluated using the agar dilution method. The overall susceptibility to fluoroquinolones has decreased rapidly after the wide use of these antimicrobial agents. Isolates of Escherichia coli and Morganella morganii proved quite susceptible to the fluoroquinolones until 1996. By 1996-97, 20% of the bacteria had become resistant to the fluoroquinolones. Activity against Klebsiella pneumoniae and Proteus mirabilis remained excellent, however, with more than 90% of the isolates susceptible during the survey period. Interestingly, some strains of Serratia marcescens and Pseudomonas aeruginosa were demonstrating in vitro resistance even before the fluoroquinolones were launched in Taiwan. This resistance increased rapidly, with around 20-30% of S. marcescens isolates fluoroquinolone-resistant by 1996-1997. After the introduction of the fluoroquinolones, resistance was demonstrated for about 15% of the P. aeruginosa isolates, and 20% of Enterobacter cloacae isolates, with the susceptibility rate remaining steady state through the 1990s. Different pulsed field gel electrophoresis patterns have been demonstrated for most fluoroquinolone-resistant isolates, suggesting that the increased resistance was not due to the spread of a single clone. We conclude a trend of increasing fluoroquinolone resistance for many common gram-negative bacteria in Taiwan, especially in recent years. Appropriate use of the new fluoroquinolones should be encouraged in order to prevent the rapid emergence and increase of fluoroquinolone-resistant bacteria. PMID- 12088624 TI - Antimicrobial susceptibility patterns of beta-hemolytic and viridans group streptococci: report from the SENTRY Antimicrobial Surveillance Program (1997 2000). AB - Susceptibility patterns of 15 antimicrobial agents were assessed for 3,400 isolates of beta-hemolytic (betahS) and viridans group (VgS) streptococci in the four regions of the SENTRY Antimicrobial Surveillance Program: Asia-Pacific (APAC), Europe (EU), Latin America (LA) and North America (NA). In 1997 through 2000, SENTRY Program monitors tested strains by reference broth microdilution methods and results were interpreted using National Committee for Clinical Laboratory Standards criteria. Among the betahS processed, 81.9% of strains were either Streptococcus pyogenes (n = 650) or S. agalactiae (n = 1,190). The VgS were generally classified as unspeciated alpha-hemolytic streptococci (n = 512; 44%) or S. mitis (n = 254; 22%). Seven quinolones, two beta-lactams, erythromycin (ER), clindamycin (CM), quinupristin/dalfopristin (Q/D), vancomycin (VA), teicoplanin (TP) and linezolid (LZ) were tested. Rank order of susceptibility for betahS isolates was: ceftriaxone (CTX) = Q/D = VA = TP = LZ (100.0%) > gatifloxacin (GATI) = trovafloxacin (TROV, 99.8%) > levofloxacin (LEVO; 99.7%) > penicillin (PEN; 99.3%) > grepafloxacin (GREPA; 97.4%) > CM (94.4%) > ER (85.5%). ER versus betahS had the highest MIC(90) values (2 microg/ml) and the lowest susceptibility rates across all regions (range, 81.4% in NA to 97.3% in LA). Among the VgS, susceptibility rank order was: VA = TP = LZ (100.0%) > Q/D (99.1%) > GATI = LEVO = TROVA (98.0%) > GREPA (96.5%) > CTX (92.8%) > CM (90.3%) > PEN (68.6%) > ER (64.5%). Macrolide resistance in both streptococcal species groups of the M-phenotype was highest in the Americas, with erm-patterns predominating in EU and APAC regions. BMS284756 among the monitored new agents showed a four- to eight-fold greater potency versus these streptococcal isolates when compared to the other six tested quinolones. Like Streptococcus pneumoniae, these other streptococci appear to have acquired numerous resistances and require continued surveillance to direct adequate therapies. PMID- 12088623 TI - In vitro bactericidal activity of piperacillin, gentamicin, and metronidazole in a mixed model containing Escherichia coli, Enterococcus faecalis, and Bacteroides fragilis. AB - An anaerobic, mixed model assay was used to study the bactericidal activities of piperacillin, gentamicin, and metronidazole, alone and in double- and triple antibiotic combinations against a polymicrobial suspension of E. coli, E. faecalis, and B. fragilis. Only slight differences were noted with the agents when tested against single (10(5) cfu/mL inoculum) versus polymicrobic suspensions (10(6) cfu/mL final inoculum) of susceptible and resistant organisms. Contrary to previous reports in the literature, metronidazole was not active against E. coli in an anaerobic environment (even in the presence of B. fragilis) nor was the activity of metronidazole reduced against B. fragilis in the presence of E. faecalis. Gentamicin demonstrated excellent activity against E. coli when tested in a Bactron anaerobic chamber (5% hydrogen, 5% CO(2,) 90% nitrogen). The pH of the media was only reduced to 6.3-6.7, considerably higher than the pH range of 5-6 needed to significantly reduce the activity of aminoglycosides. PMID- 12088625 TI - Azithromycin treatment failure in community-acquired pneumonia caused by Streptococcus pneumoniae resistant to macrolides by a 23S rRNA mutation. AB - In this report, we describe an azithromycin treatment failure in community acquired pneumonia. During the first three days of azithromycin, the patient's symptoms worsened, and she was subsequently admitted to the hospital. Blood cultures were positive for a penicillin-susceptible, macrolide-resistant S. pneumoniae. DNA sequencing revealed an A2059G mutation in domain V of the 23S rRNA. To our knowledge, this is the first clinical report of an azithromycin failure in the treatment of S. pneumoniae resistant to macrolides by this mechanism. PMID- 12088627 TI - Is serological diagnosis of Legionnaires' disease a reliable method? PMID- 12088626 TI - Nosocomial fungemia due to Trichosporon asteroides: firstly described bloodstream infection. AB - Trichosporon spp. are oppurtunistic yeasts that cause deep-seated, mucosa associated, and superficial infections in immunocompromised patients. It is well known that Trichosporon asteroides is mainly responsible of superficial infections and does not cause systemic infections in humans so far. In this study, we present the first case of disseminated infection due to Trichosporon asteroides in an intensive care patient. Yeast colonies were isolated from the specimens of blood, urine, aspiration fluid of the endotracheal tube and catheter tip swabs of the patient. Conventional mycological studies were not adequate for the identification of the isolate to the species level. The genetic identification of the yeast isolate was performed and the DNA sequence of the isolate exactly matched the corresponding sequence of the Trichosporon asteroides rRNA gene from the GenBank DNA database (accession numbers: AB018017, AF075513). Therefore, our isolate was identified as Trichosporon asteroides as a causative agent of deep-seated fungemia. PMID- 12088630 TI - Biocomplexity: the post-genome challenge in ecotoxicology. AB - There are four crucial challenges for the environmental toxicologists in the next decade: (1) understanding the mechanisms of molecular and subcellular interactions with pollutant chemicals, including genomic and proteomic aspects; (2) the development of predictive simulation models of toxic effects on complex cellular and physiological processes; (3) linking molecular, cellular and patho physiological 'endpoints' with higher level ecological consequences; and (4) precautionary anticipation of possible harmful impacts of novel developments in industrial processes, including biotechnology and nanotechnology. One of the major difficulties in ecotoxicology is to link harmful effects of chemical pollutants in individual animals and plants with the ecological consequences. Consequently, this obstacle has resulted in a 'knowledge-gap' for those seeking to develop policies for sustainable use of resources and environmental protection. The overall problem is: how to develop effective procedures for environmental/ecological impact and risk assessment? However, the use of diagnostic 'clinical-type' tests or 'biomarkers' has started to provide information on the health-status of populations based on relatively small samples of individuals. Also, biomarkers can now be used to begin to link processes of molecular and cellular damage through to the higher levels (i.e. prognostic capability), where they can result in reduced performance and reproductive success. Research effort to meet this challenge must be inter-disciplinary in character, since the key questions mainly involve complex interfacial problems. These include effects of physico-chemical speciation on uptake and toxicity, the toxicity of complex mixtures; and linking the impact of pollutants through the various hierarchical levels of biological organisation to ecosystem and human health. Finally, the development and use of process-based computational simulation models (i.e. 'virtual' cells, organs and animals), illustrated using an endosomal/lysosomal uptake and cell injury model, will facilitate the development of a predictive capacity for estimating risk associated with the possibility of future environmental events. PMID- 12088631 TI - Sex-specific biotransformation and detoxification after xenobiotic exposure of primary cultured hepatocytes of European flounder (Platichthys flesus L.). AB - Sex-specific effects of sublethal concentrations of known effective pro-oxidants such as 100,200 and 400 microM benzo[a]pyrene (B[a]p), 50 microM nitrofurantoin (NF) and 100 microM hydrogen peroxide (H2O2) on biotransformation pathways were studied in isolated hepatocytes of immature female and male European flounder (Platichthys flesus L.). Cell responses were assessed at the level of: (1) stress induction as measured by formation of reactive oxygen species (ROS), mainly superoxide radicals, and induction of cytochrome P450 (CYP450) biotransformation activity; (2) cellular antioxidant defences, both non-enzymatic (reduced glutathione) and enzymatic (DT-diaphorase (DTD) or quinone oxidoreductase, EC 1.6.99.2); (3) detoxification (aldehyde dehydrogenase (ALDH), EC 1.2.1.3); and (4) cellular damage as measured by reduced lysosomal membrane stability and cell death. As there is increasing evidence that 17-beta-estradiol interferes with certain pathways of xenobiotic biotransformation, we additionally tested the effects of different concentrations of 17-beta-estradiol (0.2-10 microM) alone and 17-beta-estradiol (1 microM) in combination with 100 microM B[a]p. Parameters were monitored after 1 and 9 days of exposure by quantitative image analysis of chromogenic or fluorogenic reaction products. Our study revealed sex-dependent differences in cellular stress responses. In hepatocytes of female flounder, biotransformation was slower and the capacity of non-enzymatic antioxidant defences and detoxification of toxic aldehydes was lower than in males. Additional administration of 17-beta-estradiol enlarged these differences between the sexes with respect to biotransformation activity and antioxidant defence in xenobiotic-induced injury. These findings may explain the higher susceptibility of female flounder to toxic and carcinogenic compounds in the marine environment. PMID- 12088632 TI - Effects of chronic dietary exposure to environmentally relevant concentrations to 2,3,7,8-tetrachlorodibenzo-p-dioxin on survival, growth, reproduction and biochemical responses of female rainbow trout (Oncorhynchus mykiss). AB - Adult female rainbow trout were exposed to dietary 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) at concentrations of 1.8, 18 and 90 ng TCDD/kg (ww) food for up to 300 day. At the end of the exposure fish were spawned and the reproductive outcomes were assessed. TCDD was accumulated into tissues and eggs in a dose dependent manner with steady state being achieved after 50-100 day of exposure. Biochemical and hematological parameters were monitored at 50, 100, 150, and 200 day after the beginning of exposure. The survival of adult female trout was reduced in a dose-dependent manner by exposure to TCDD in the diet. Fish fed 1.8 ng TCDD/kg, moist weight of diet, showed significantly reduced survival compared with those fed the control diet. TCDD also affected survival of fry from females fed 1.8 ng TCDD/kg. Observed adverse effects in adult fish were as sensitive as early life-stage endpoints. Liver EROD activity was only moderately increased in all exposure groups after 250+ day of exposure. Low rates of edema and deformities were observed in fry from all treatment groups including controls. This study has demonstrated adverse effects of TCDD to both adults and fry at concentrations comparable to current environmental concentrations. This suggests that direct adult toxicity as well as reproductive endpoints need to be incorporated in the current risk assessment paradigm for these compounds. PMID- 12088633 TI - Swimming and ammonia toxicity in salmonids: the effect of sub lethal ammonia exposure on the swimming performance of coho salmon and the acute toxicity of ammonia in swimming and resting rainbow trout. AB - This study tested the hypothesis that swimming exacerbates ammonia toxicity in fish. Both sub-lethal and acute toxicity testing was conducted in a swim tunnel on swimming and resting coho salmon and rainbow trout, respectively. The sub lethal tests on coho salmon also considered the compartmentalization of ammonia within the fish. Coho salmon showed a significant linear decrease in U(crit) both with increasing water ammonia (0, 0.02, 0.04 and 0.08 mg per l NH3) and increasing plasma ammonia. Data collected included plasma pH and ammonia, muscle pH and ammonia and muscle membrane potential. Based on results found in these experiments it was concluded that the reduction in swimming performance was due to both metabolic challenges as well as depolarization of white muscle. Acute toxicity testing on swimming and resting rainbow trout revealed that swimming at (60% U(crit) or approximately 2.2 body lengths/s) decreased the LC50 level from 207+/-21.99 mg N per l in resting fish to 32.38+/-10.81. The LC50 for resting fish was significantly higher than that for swimming fish. The acute value set forth by the US EPA at the same pH is 36.1 mg N per l and may not protect swimming fish. In addition the effect of water hardness on ammonia toxicity was considered. It was found that increased water calcium ameliorates ammonia toxicity in fish living in high pH water. PMID- 12088634 TI - The effect of feeding and fasting on ammonia toxicity in juvenile rainbow trout, Oncorhynchus mykiss. AB - Present fresh water ammonia standards have been established using data collected from toxicity tests on unfed fish. Ammonia, however, is an unusual toxicant as it is produced as a metabolic waste following protein catabolism. The present research was conducted to investigate the relationship between feeding and ammonia toxicity in rainbow trout, Oncorhynchus mykiss. Results from these studies revealed that some fish fed to satiation have plasma ammonia levels greater than 30 microg/ml. This level was similar to the plasma ammonia levels in rainbow trout at the ammonia LC50 value calculated in the present experiments. Even though plasma ammonia in fed fish was elevated there was no significant difference between the 96 h LC50 values for fed and unfed fish (174 mg N per l) at pH 7.2. Feeding rates during these experiments decreased during the first 48 h of ammonia exposure, but increased again in the second 48 h at all but the highest ammonia level. Feeding rate never increased to the control level in ammonia exposed fish. In a second set of experiments feeding fish had a significantly higher 24 h LC50 level, 177 mg N per l, than fish fasted for 5 or 10 days, 135-143 mg N per l. No significant difference was noted however, between the 48 h LC50 values for fed and fasted fish. It was evident from these studies that feeding protects rainbow trout from ammonia toxicity during the first 24 h of exposure and that fasting exacerbates ammonia toxicity. PMID- 12088635 TI - Effects of exposure of Mya arenaria and Mactromeris polynyma to contaminated marine sediments on phagocytic activity of hemocytes. AB - Two species of bivalves, Mya arenaria and Mactromeris polynima, were exposed to contaminated marine sediments from Baie des Anglais, Quebec, for a period of 10 and 12 weeks, respectively, in order to determine if there was an effect on the phagocytic activity of hemocytes from each species. These sediments contain elevated levels of both PAHs and PCBs. Uncontaminated beach sand was used as control sediments. After a period of 4 weeks, each species of bivalves were sampled and hemocyte phagocytic activity was monitored by flow cytometry. While phagocytosis by hemocytes from M. polytiyma was significantly suppressed, those from M. arenaria were not different from beach sand-exposed controls. At the end of the exposure period, the phagocytic activity of hemocytes from both species was suppressed. Physiological parameters such as mantle proteins or malondialdehyde levels, total protein and total glycogen levels in the digestive gland were not affected by exposure to contaminated sediments. Moreover, the suppression of phagocytosis was well correlated with the transfer of contaminants from the sediments to the bivalves and their subsequent bioaccumulation, as demonstrated by the PCB body burden. These results support the use of bivalves as good sentinel species to survey sediment contamination and the usefulness of hemocyte phagocytic activity as a sensitive biomarker of exposure to organic contaminants. PMID- 12088636 TI - Reproductive alterations in adult grass shrimp, Palaemonetes pugio, following sublethal, chronic endosulfan exposure. AB - Grass shrimp (Palaemonetes pugio) populations exposed to anthropogenic contaminant sources in South Carolina (SC) have reduced densities when compared with populations at SC-reference sites. This laboratory study examined the effects of a commonly used agricultural insecticide, endosulfan, on grass shrimp reproduction. Reproductively active grass shrimp were chronically exposed to sublethal concentrations of endosulfan (200 or 400 ng/l) for 43 days. The cumulative number of females that became gravid and the rate at which they became gravid were measured. Endosulfan exposure reduced the cumulative number of gravid females by 31% in the 200 ng/l exposure and 39% in the 400 ng/l exposure. The first appearance of gravid females in the population was significantly delayed in treated populations compared with the control treatment in a dose dependent manner. Clutch size in these gravid females was not significantly different among the treatments. Additionally, there was no difference in the onset of reproduction in the treated populations. These results implicate a population reduction due to a decrease in the overall number of females becoming gravid in a population over time, not a reduction in clutch size per individual. While the mechanisms of action have yet to be defined, these results indicate that sublethal endosulfan concentrations may have a negative effect on grass shrimp reproductive biology. PMID- 12088637 TI - Evaluating molar CYP1A level in fish hepatic microsomes by competitive ELISA using recombinant membrane-free CYP1A standard protein. AB - Fish cytochrome P4501A (CYP1A) is a widely accepted environmental biomarker, detecting biological effects of several xenobiotic groups present in aquatic environments, when evaluated in target tissues of a biosensor species. However, appropriate utilization of its protein level as a routine environmental diagnostic tool requires evaluation of properly normalized molar levels, mitigating comparison among different laboratories, during a multi-annual time scale and over a variety of tested populations of the biosensor species. A competitive enzyme-linked immunosorbent assay (ELISA) was developed for determination of CYP1A of the striped sea bream, Lithognathus mormyrus, using our previously described antibody raised to a trout CYP1A synthetic peptide, and a recombinant L. mormyrus CYP1A as a competitor. The L. mormyrus CYP1A-cDNA was cloned and modified by truncating its 5' hydrophobic membrane anchor, as well as by addition of 4x histidine tag, permitting its partial purification on a nickel NTA column. The modified cDNA was ligated into the PCWOri+ vector and heterologously produced in Escherichia coli as a cytosolic, membrane-free protein, retaining its immuno-affinity with the anti-CYP1A antibody in the presence of the detergent Triton X-100. The detergent was added to the ELISA solution during the competitive step, rendering the microsomal CYP1A more accessible to the antibody. ELISA components, including coated levels of the modified standard CYP1A, and the concentrations of the Triton X-100, CYP1A specific antibody, and the range of dissolved CYP1A standard protein, were optimized. Hypothesized immuno-affinity differences between the microsomal and the recombinant CYP1As, and among microsomal samples, as well as assay accuracy, were examined and discussed. This ELISA can serve for more efficient utilization of fish CYP1A as a pollution biomarker, and also as a model for establishing competitive ELISAs aimed at quantification of many different microsomal P450 proteins. PMID- 12088638 TI - Bioaccumulation of trace metals in the copepod Calanoides acutus from the Weddell Sea (Antarctica): comparison of two-compartment and hyperbolic toxicokinetic models. AB - Bioaccumulation of Cd, Co, Cu, Ni, Pb and Zn in the Antarctic calanoid copepod Calanoides acutus (Giesbrecht, 1902) was investigated during a cruise of RV 'Polarstern' to the Weddell Sea. Main goals were to provide information on accumulation strategies of the organisms tested and to verify toxicokinetic models as a predictive tool. Except for Cd, the organisms accumulated metals upon exposure. It was possible to estimate significant model parameters of two compartment and hyperbolic models. These models were successfully verified in a second toxicokinetic uptake study. The model verification was extended in a third uptake study with increasing external metal dosing. We found a linear increase of net uptake with external waterborne metal exposures up to 80 microg Pb l(-1), with excellent predictions of the two-compartment model. For Co both models give reasonable predictions up to 20 microg Co l(-1). Regarding Cu, Ni and Zn only hyperbolic model predictions were in good agreement with measured values up to 150 microg Cu l(-1), 80 microg Ni l(-1) and 290 microg Zn l(-1). Due to a decrease of Cd body burden in the experiments, only the hyperbolic model was applicable, leading to reliable predictions up to 20 microg Cd l(-1). These concentrations largely determine the range for which these models may serve as a predictive tool. PMID- 12088639 TI - Keratinocytes and cytokines. AB - The skin has long been recognized as a major producer of cytokines, but the keratinocyte as principal epidermal cell has received less attention as potential source and target of cytokines. Nevertheless, keratinocytes produce a plethora of cytokines including interleukin (IL)-1, -6, -7, -8, -10, -12, -15, -18, and -20, and tumor necrosis factor alpha (TNF). The production by keratinocytes of pro inflammatory (IL)-1, -6, -8, and TNF was recognized early and is well studied. Keratinocyte-derived IL-7 and -15 are considered to be significant in T-cell trafficking, possibly even in the pathogenesis of cutaneous T-cell lymphoma. Immunomodulatory IL-10 and -12 originating from keratinocytes are considered to be responsible for systemic effects, and IL-18 perhaps has a similar action. Keratinocytes were fairly recently recognized as being source or target of other IL-10 family members like IL-20 and IL-24 and the role of these cytokines in specific diseases is under investigation. In addition, a variety of cytokine receptors are present on keratinocytes like those for IL-4, -13, and -17 and to lesser degree IL-2. The ability to study the expression of cytokines by keratinocytes in vivo and in vitro using primary cells, immortalized cells or even organotypic culture systems offers many possibilities to further investigate the role of cytokine production in keratinocyte biology and disease. PMID- 12088640 TI - Evaluation of a specific immunochemotherapy for the treatment of canine visceral leishmaniasis. AB - The efficacy of specific immunochemotherapy against Leishmania infantum infection in dog was studied. The effects on transmission of the disease, as well as the cellular and humoral immune response were examined. The treated animals showed a significant reduction in the infection rates that were detected in Phlebotomus perniciosus females fed on the dog. The humoral immune response, assayed with an indirect immunofluorescence antibody test (IFAT), did not show significant variations under the influence of the therapy. The characterisation of the peripheral blood mononuclear cells (PBMC) using flow cytometry indicated a significant increase in the proportion of T lymphocytes, especially of CD4/TcR(alpha)(beta)(+) and CD4/CD45RA(+) cells, without showing evidence for modifications in the other leukocyte subsets. Cellular lymphoproliferation studies indicated a lack of a specific response to soluble leishmanial antigen (SLA), but the non-specific lymphoproliferative capacity assayed with phytohemagglutinin (PHA) was maintained. PMID- 12088642 TI - Production and characterization of monoclonal antibodies against dog immunoglobulin isotypes. AB - A panel of six monoclonal antibodies (mAbs) recognizing antigenic determinants on canine immunoglobulin (Ig) heavy or light chains was produced and characterized. All monoclonals recognized the IgG(2) subclass, although only two were subclass specific (CA3H1 and CA4F1). The CA3B8 mAb was found to be specific for an epitope on canine immunoglobulin G heavy chain, (IgG(1) and IgG(2) subclasses). Two mAbs (CA2E9 and CA5B2) reacted with an epitope on the heavy chain of canine IgG and IgM and another, CA4E7, bound to canine IgA, IgG and IgM isotypes; CA4E7 recognized an epitope on canine immunoglobulin light chain. CA4E7, CA4F1 and CA5B2 recognized an epitope in the Fab region. Three mAbs, CA3B8, CA4E7 and CA5B2, showed much lower reactivity with canine IgG by ELISA when IgG was periodate-treated, suggesting that they recognized a carbohydrate determinant. Cross-reactivity analysis of these mAbs with sera from horse, goat, cow, sheep, pig, cat, rabbit, hamster, rat, mouse and human indicated that two mAbs, CA3B8 and CA5B2, recognized a canine IgG-specific epitope; two others, CA3H1 and CA4E7, recognized an epitope also present in rabbit and sheep immunoglobulin respectively; and the remaining two (CA2E9 and CA4F1) recognized an epitope broadly present on the Igs of the species analyzed. This panel of antibodies will be a useful tool for future canine immunodiagnosis tests. With the exception of CA2E9, all mAbs were able to recognize plasma cells on paraffin-embedded tissues, and will thus be useful for immunohistochemical assays. PMID- 12088641 TI - Cytokine expression during the outcome of canine experimental infection by Leishmania infantum. AB - In this study, the cytokines, interferon-gamma (IFN-gamma), interleukin-2 (IL-2), IL-12 p40, IL-6 and IL-10, expressed by peripheral blood mononuclear cells of 13 beagle dogs inoculated with Leishmania infantum amastigotes, were analysed during a period of up to 23 months. The course of infection was monitored through clinical and parasitological examinations, haematological alterations and serum antileishmania antibody levels. Dogs developed symptomatic infections with haematological alterations, humoral immune response and reduced specific lymphoproliferative response. Parasite presence was detected in bone marrow, popliteal lymph node and skin. Specifically stimulated cytokine transcripts were generally observed in a low proportion of dogs, except at months 9, 10 and 11 post-infection where there was a considerable increase in the proportion of dogs expressing IFN-gamma and IL-2 mRNA. IL-12 p40 and IL-10 transcripts were sporadically detected in few animals. In non-infected animals, IFN-gamma mRNA was the only detectable cytokine but only in cells cultured in the presence of concanavalin A (ConA). The low proportion of animals expressing specific cytokines, during the first 8 months of infection associated with evidences of parasite dispersion without clinical signs of disease, suggests the occurrence of a relatively "silent establishment" of the parasite avoiding adverse host-cell mediated immunological reactions. The humoral immune response displayed in these animals, the cell-mediated immunosuppression, nor the disease severity could be related with the expression of IL-10. The predominance of a Th1 type response for a relatively short period indicates that these cytokines are required to control the infection delaying the appearance of progressive disease. PMID- 12088643 TI - Cloning of bovine CD69. AB - CD69 is rapidly inducible on various hematopoietic cells upon stimulation and is detectable as an early activation antigen. Although CD69 is well characterized in human and mouse, no information is available on bovine CD69. We report here that, bovine CD69 was cloned from a cDNA expression library prepared from activated peripheral blood lymphocytes. The full-length cDNA contained an 80bp 5' untranslated region, followed by a 600bp coding region and AU-rich motifs in a 3' untranslated region (GenBank accession number AF272828). Comparison of the bovine CD69 coding sequence reveals 69.4 and 78.2% nucleotide sequence identities with mouse and human CD69, respectively. The predicted amino acid sequence of bovine CD69 shares 56.3 and 62.3% sequence identity when compared with mouse and human CD69, respectively. Bovine CD69 has the highly conserved amino acid sequences found in the C-type lectin family, suggesting that the conserved residues may be important for conformation and binding to the, as yet unidentified ligand. In addition, the cytoplasmic tail of bovine CD69 has two casein kinase-2 (CK-2) phosphorylation sites. These data suggest that bovine CD69 plays an important role in the activation of lymphocytes. PMID- 12088644 TI - Development and characterization of monoclonal antibodies to chicken interleukin 15. AB - The chicken IL-15 gene was recently cloned and shown to encode a polypeptide with T cell growth factor activity similar to IL-2. To further characterize the chemical and biological properties of chicken IL-15, we generated a panel of monoclonal antibodies against bacterially expressed protein and characterized their binding specificities. All antibodies were reactive by ELISA with recombinant IL-15, but not IL-2, and identified a 15kDa recombinant chicken IL-15 by Western blot analysis. Two antibodies inhibited IL-15-induced proliferation of splenic lymphoblast cells. These monoclonal antibodies will be useful for further structural and immunological studies of chicken IL-15. PMID- 12088645 TI - Specific mucosal IgA immunity in turkey poults infected with turkey coronavirus. AB - The objective of this study was to elucidate the kinetics and magnitudes of specific IgA antibody responses in intestines of turkey poults infected with turkey coronavirus (TCV). Turkey poults were orally inoculated with TCV at 10 days of age. Intestinal segment cultures were administered for duodenum, jejunum, and ileum and the IgA antibody responses were analyzed at 1, 2, 3, 4, 6, or 9 weeks post-infection (PI) in two different experiments. The kinetics of virus specific IgA antibody responses in duodenum, jejunum, and ileum were similar: gradually increased from 1 week PI, reached the peak at 3 or 4 weeks PI, and declined afterward. The virus-specific IgA antibody responses in duodenum, jejunum, and ileum showed negative correlation with duration of TCV antigen in the corresponding locations of intestine with Spearman's correlation coefficient of -0.85 (p=0.034), -0.74 (p=0.096), and -0.75 (p=0.084), respectively. Moreover, the virus-specific IgA antibody responses in serum were positively correlated with that of duodenum (coefficient=0.829, p=0.042), jejunum (coefficient=0.829, p=0.042), and ileum (coefficient=0.771, p=0.072) segment cultures, suggesting that the induction of specific IgA response in serum was predictive of an IgA response in intestine. The results indicate that intestinal mucosal IgA antibodies to TCV are elicited in turkeys following infection with TCV. The local mucosal antibodies may provide protective immunity for infected turkeys to recover from TCV infection. PMID- 12088646 TI - The effects of cAMP modulation upon the adhesion and respiratory burst activity of immune complex-stimulated equine neutrophils. AB - Toxic products such as reactive oxygen intermediates released by activated polymorphonuclear neutrophil (PMN) have an important role in the pathophysiology of diseases associated with the deposition of immune complexes (IC) in tissues. IC-induced activation of PMN requires adhesion mediated by integrin adhesion receptors. Of the integrins expressed on PMN, the beta(2) family has been found to be of particular importance for activation of PMN by IC. beta(2) Integrin ligand binding must be activated to enable adhesion to IC. Both activating and inhibitory signals regulate beta(2) integrin ligand avidity and adhesion. The second messenger cyclic adenosine monophosphate (cAMP) has been demonstrated to inhibit the activation of PMN in response to a variety of stimuli. The purpose of this study is to test the hypothesis that cAMP-dependent signals inhibit beta(2) integrin-dependent adhesion of equine PMN to immobilized IC and subsequent adhesion-dependent activation of respiratory burst activity. Treatment of equine PMN with beta(2) adrenergic agonists isoproterenol or clenbuterol, which trigger an increase in intracellular cAMP concentration, inhibited adhesion of equine PMN to IC in a dose dependent manner. Similarly, inhibition of cAMP hydrolysis by the non-specific phosphodiesterase (PDE) inhibitor pentoxifylline and the PDE 4 specific inhibitor rolipram inhibited adhesion of equine PMN to IC. Elevation of intracellular cAMP levels with pentoxifylline, clenbuterol and rolipram also inhibited IC-induced activation of respiratory burst activity in equine PMN. Importantly, co-treatment of equine PMN with rolipram and either beta(2) adrenergic agonist synergistically inhibited both the adhesion of equine PMN to IC as well as the subsequent respiratory burst activity. PMID- 12088647 TI - Lesional expression of thymus and activation-regulated chemokine in canine atopic dermatitis. AB - In this study, we investigated the mRNA expression of a chemokine, thymus and activation-regulated chemokine (TARC), and cytokines including IL-1beta, IL-4, IFN-gamma and TNF-alpha in skin samples obtained from both dogs with atopic dermatitis (AD) and healthy dogs. TARC mRNA was found to be selectively expressed in lesional skin of the dogs with AD, but not in non-lesional skin of the dogs with AD or the normal skin of the healthy dogs. The expression levels of IL 1beta, IFN-gamma and TNF-alpha in the lesional skin were also significantly higher than those in the non-lesional skin of the dogs with AD. However, IL-4 mRNA was not detected in any of the skin samples in this study. The present results suggest that TARC and inflammatory cytokines such as IL-1beta, IFN-gamma and TNF-alpha may play roles in the pathogenesis of canine AD as well as that of human AD. PMID- 12088648 TI - Characterisation of antisera to recombinant IgA of the common brushtail possum (Trichosurus vulpecula). AB - One of the limiting factors in understanding immune responses in marsupials is the scarcity of marsupial specific immunological reagents. This paper describes the characterisation of an antiserum raised against a recombinant protein of the constant region of the heavy chain of IgA (C(alpha)) of the common brushtail possum (Trichosurus vulpecula). The availability of a marsupial specific anti-IgA provides a useful tool for the characterisation of mucosal immune responses in possums. Anti-C(alpha) specifically detects IgA in possum serum and secretions using ELISAs, immuno-dot blots and Western blots without any cross-reactivity to IgG. The possum anti-C(alpha) cross-reacts with IgA of koala (Phascolarctos cinereus), tammar wallaby (Macropus eugenii) and eastern grey kangaroo (Macropus giganteus), demonstrating the potential for use in other marsupials. PMID- 12088649 TI - IgG antibody subclass response against equine herpesvirus type 4 in horses. AB - In this study, IgG subclass responses against equine herpesvirus type 4 (EHV-4) were examined by enzyme-linked immunosorbent assay (ELISA) using a type-specific region of EHV-4 glycoprotein G (gG). ELISA using sera collected from horses experimentally infected with EHV-4 revealed that IgGa and IgGb antibodies were detected at high level, but IgGc and IgG(T) antibody responses were detected at low level or were undetectable. The IgGa antibody response reached its peak on day 10 post-infection, and then dropped. The IgGb antibody response reached its maximum level on day 12 post-infection, and then the level was sustained during at least 28 days after infection. Forty healthy racehorses that had already been infected with EHV-4 possessed antibody against EHV-4. Although IgGa antibodies specific for EHV-4 were not detected in any horses, IgGb antibodies were detected and the levels correlated with total IgG antibodies against EHV-4 gG. The results suggest that EHV-4-specific IgGa and IgGb antibodies are induced in EHV-4 infected horses, and that IgGb antibody, but not IgGa, is long lasting. PMID- 12088650 TI - Maximum shields: the assembly and function of the bacterial spore coat. AB - Spores produced by bacilli and clostridia are surrounded by a multilayered protein shell called the coat. As the armor-like appearance of the coat suggests, this structure, along with others within the spore, confers the remarkable resistance properties that make Bacillus anthracis spores such potent biological weapons. Here, I review recent studies of coat assembly in the model organism Bacillus subtilis, and explore the implications of these findings for coat assembly in B. anthracis and for defense against biological weapons. PMID- 12088651 TI - To sialylate, or not to sialylate: that is the question. AB - Most oropharyngeal pathogens express sialic acid units on their surfaces, mimicking the sialyl-rich mucin layer coating epithelial cells and the glycoconjugates present on virtually all host cell surfaces and serum proteins. Unlike the host's cells, which synthesize sialic acids endogenously, several microbial pathogens use truncated sialylation pathways. How microorganisms regulate sialic acid metabolism to ensure an adequate supply of free sugar for surface remodeling is a new area of research interest to basic scientists and those focused on the clinical outcome of the host-pathogen interaction. PMID- 12088652 TI - How does Legionella pneumophila exit the host cell? AB - In recent years, tremendous progress has been made in unraveling the elegant mechanisms by which intracellular pathogens invade host cells and establish intracellular infections. By contrast, our knowledge of the mechanisms of host cell cytolysis and the egress of intracellular pathogens is still in its infancy. Temporal pore-formation-mediated lysis of the host and exit by Legionella pneumophila and Leishmania could provide a new model of egress for other intracellular pathogens, many of which exhibit pore-forming or cytolysin activity PMID- 12088653 TI - Tales of the unexpected. PMID- 12088659 TI - Are autotrophs less diverse than heterotrophs in marine picoplankton? PMID- 12088660 TI - Two different open reading frames named slyA in the E. coli sequence databases. PMID- 12088662 TI - Do plant and human pathogens have a common pathogenicity strategy? AB - Recently, a novel 'two-step' model of pathogenicity has been described that suggests host-cell-derived vasculoproliferative factors play a crucial role in the pathogenesis of bacillary angiomatosis, a disease caused by the human pathogenic bacterium Bartonella henselae. The resulting proliferation of endothelial cells could be interpreted as bacterial pathogens triggering the promotion of their own habitat: the host cell. Similar disease mechanisms are well known in the plant pathogen Agrobacterium tumefaciens, which causes crown gall disease. There are notable similarities between the pathogenicity of A. tumefaciens leading to tumourous disease in plants and to the B. henselae triggered proliferation of endothelial cells in humans. Here, we hypothesize that this pathogenicity strategy might be common to several bacterial species in different hosts owing to shared pathogenicity factors. PMID- 12088663 TI - Cell turnover and cell tropism in HIV-1 infection. AB - Early infection with HIV-1 is dominated by CCR5-tropic (R5, non-syncytium inducing) viruses. The evolution of CXCR4-tropic (X4, syncytium-inducing) viruses occurs later in the infection and is associated with rapid disease progression. Here, we propose that the tropism of X4 viruses for naive CD4+ T cells is disadvantageous in early infection owing to the low division rate of these cells. In healthy individuals, the division rate of memory cells is nearly ten times higher than that of naive cells and thus the memory-cell tropism of R5 viruses could account for their dominance early in infection. As the division rate of naive T cells increases with CD4+ depletion, X4 viruses come to dominate in late disease. PMID- 12088664 TI - The role of mathematical modelling in the control of the 2001 FMD epidemic in the UK. AB - Mathematical models played an important role in guiding the development of the control policies in the 2001 foot-and-mouth disease epidemic in the UK. The variety of approaches that helped to guide the policy can sometimes be confusing. Here, the different modelling exercises that were developed over the course of the epidemic are reviewed, describing the difficulties in interpreting the available data and the appropriateness of the various assumptions. PMID- 12088665 TI - 2001: a year of major advances in anthrax toxin research. AB - Anthrax is caused when spores of Bacillus anthracis enter a host and germinate. The bacteria multiply and secrete a tripartite toxin causing local edema and, in systemic infection, death. In nature, anthrax is primarily observed in cattle and other herbivores; humans are susceptible but rarely affected. In 2001, anthrax spores were used effectively for the first time in bioterrorist attacks, resulting in 11 confirmed cases of human disease and five deaths. These events have underscored the need for improved prophylaxis, therapeutics and a molecular understanding of the toxin. The good news about anthrax is that several decisive discoveries regarding the toxin have been reported recently. Most notably, the toxin receptor was identified, the 3-D structures of two of the toxin subunits were solved and potent in vivo inhibitors were designed. These findings have improved our understanding of the intoxication mechanism and are stimulating the design of strategies to fight disease in the future. PMID- 12088666 TI - How bacteria could cause cancer: one step at a time. AB - Helicobacter pylori highlighted the potential for bacteria to cause cancer. It is becoming clear that chronic infection with other bacteria, notably Salmonella typhi, can also facilitate tumour development. Infections caused by several bacteria (e.g. Bartonella spp., Lawsonia intracellularis and Citrobacter rodentium) can induce cellular proliferation that can be reversed by antibiotic treatment. Other chronic bacterial infections have the effect of blocking apoptosis. However, the underlying cellular mechanisms are far from clear. Conversely, several bacterial toxins interfere with cellular signalling mechanisms in a way that is characteristic of tumour promoters. These include Pasteurella multocida toxin, which uniquely acts as a mitogen, and Escherichia coli cytotoxic necrotizing factor, which activates Rho family signalling. This leads to activation of COX2, which is involved in several stages of tumour development, including inhibition of apoptosis. Such toxins could provide valuable models for bacterial involvement in cancer, but more significantly they could play a direct role in cancer causation and progression. PMID- 12088674 TI - Balancing protective immunity and immunopathology. AB - The immune system is fighting a constant war against pathogens in its own territory. This requires a potent arsenal for efficient control of pathogens but also requires tight regulatory mechanisms in order to avoid excessive collateral damage. Maintaining equilibrium is the daily challenge of the immune system. Interactions between pathogens, antigen-presenting cells (APCs) and lymphocytes are critical in this balancing act. Of particular importance for the generation of protective immune responses is the induction of activation programs in APCs directly by pathogens or by T cell derived factors, such as CD40L, RANKL or cytokines. In order to counterbalance overshooting immune responses, T cells and APCs secrete anti-inflammatory cytokines that are key for maintaining a healthy balance between protection and immunopathology. PMID- 12088675 TI - How dendritic cells and microbes interact to elicit or subvert protective immune responses. AB - B and T lymphocytes recognize antigens with high specificity, but neither initiate immune responses, nor decide their types. These functions rest upon dendritic cells (DCs), which can determine and maintain Th1/Th2 polarization. Immune responses are thus dependent on the DC subset, the receptors that recognize each pathogen and the microenvironment. Microbes employ an array of mechanisms to evade and disrupt DC functions; some even hijack DCs for transport around the body. Our progress in the understanding of DC physiology will hopefully help us create the necessary vaccines to counteract the infectious agents that still plague mankind. PMID- 12088676 TI - Links between innate and adaptive immunity via type I interferon. AB - Type I interferon (IFN-alpha/beta) is expressed rapidly following exposure to a wide variety of infectious agents and plays a key role in innate control of virus replication. Recent studies have demonstrated that dendritic cells both produce IFN-alpha/beta and undergo maturation in response to IFN-alpha/beta. Moreover, IFN-alpha/beta has been shown to potently enhance immune responses in vivo through the stimulation of dendritic cells. These findings indicate that IFN alpha/beta serves as a signal linking innate and adaptive immunity. PMID- 12088677 TI - The role of dendritic cells during Salmonella infection. AB - One type of phagocytic antigen-presenting cell (APC) - the dendritic cell (DC) - may have specialized functions during infection with the bacterium Salmonella, including a possible role in transporting Salmonella across the intestinal barrier. In addition, changes in the number, localization and cytokine production of CD8alpha+, CD8alpha-CD4+ and CD8alpha-CD4- DC subsets occur during infection. DCs function in stimulating bacteria-specific T cells by direct presentation of Salmonella antigens and as bystander APCs. Studying the function of DCs during Salmonella infection provides insight into the capacity of these sophisticated APCs, which are a key link between innate and adaptive immunity, to initiate and modulate the immune response to a bacterial infection. PMID- 12088678 TI - The role of IFN-gamma in the outcome of chlamydial infection. AB - Chlamydia are intracellular bacteria which infect many vertebrates, including humans. They cause a myriad of severe diseases, ranging from asymptomatic infection to pneumonia, blindness or infertility. IFN-gamma plays an important role in defense against acute infection and in the establishment of persistence. Chlamydia have evolved mechanisms to escape IFN-gamma functions. IFN-gamma mediated effector mechanisms may involve effects on the metabolism of tryptophan or iron, on the inducible NO synthase (iNOS), on the secretion of chemokines and adhesion molecules or on the regulation of T-cell activities. IFN-gamma is secreted by the innate and the adaptive arms of the immune system. Within the former, Chlamydia-infected macrophages express IFN-gamma that in turn mediates resistance to infection. IFN-alpha/beta are pivotal for both IFN-gamma- and iNOS mediated resistance to chlamydial infection in macrophages. PMID- 12088679 TI - Control of Mycobacterium tuberculosis through mammalian Toll-like receptors. AB - An efficient immune response against the intracellular pathogen Mycobacterium tuberculosis is critically dependent on rapid detection of the invader by the innate immune response and the activation of the adaptive immune response. Toll like receptors (TLRs) contribute to innate immunity by the detection of Mycobacteria-associated molecular patterns and mediating the secretion of antibacterial effector molecules. TLRs influence the adaptive immune response by upregulation of immunomodulatory molecules supporting the development of a Th1 biased T cell response. In this manner, activation of TLRs contributes to defense against microbial infection. PMID- 12088680 TI - Malaria, monocytes, macrophages and myeloid dendritic cells: sticking of infected erythrocytes switches off host cells. AB - The adhesive phenotypes expressed by Plasmodium-falciparum-infected erythrocytes were previously thought simply to permit sequestration of parasites in the peripheral circulation. Recent work has illuminated how falciparum-infected erythrocytes may modulate the function of monocytes, macrophages and myeloid dendritic cells through the action of haemozoin from digested haemoglobin and through adhesion of infected cells to their surface. PMID- 12088681 TI - Dendritic cells and immunity to leishmaniasis and toxoplasmosis. AB - There is increased recognition that dendritic cells (DCs) are an important source of the IL-12 required to initiate protective immunity to protozoa, such as Leishmania and Toxoplasma. This article reviews the advances made in the last two years in understanding the pathways that lead to DC activation after infection with these organisms. Interestingly, there appear to be differences in the DC activation pathways utilized by these two intracellular protozoa which also may differ from the pathways utilized by bacteria. PMID- 12088682 TI - Interactions between dead cells and dendritic cells in the induction of antiviral CTL responses. AB - Dendritic cells - professional antigen-presenting cells - are key players for activating adaptive immune responses against viruses. Apoptosis or lytic cell death often accompanies viral infection. Dendritic cells can acquire infected dead or dying cells as exogenous sources of antigens for presentation on MHC class I and II molecules to initiate T cell responses. This pathway of activating T cells may be critical for the development of effective antiviral immunity in vivo. PMID- 12088684 TI - Promises and pitfalls in the reconstitution of immunity in patients who have HIV 1 infection. AB - Recent studies suggest that immunologic containment of HIV-1 is possible and the immune correlates of this control are being defined. There have been significant advances in our ability to elicit virus-specific immune responses, which have led to viral control in animal models. Although viral escape from the immune system is an important challenge, new strategies for therapeutic and preventative HIV-1 vaccination are being vigorously pursued in human trials. PMID- 12088683 TI - Avoiding the kiss of death: how HIV and other chronic viruses survive. AB - Virus-specific CD8 T cells during chronic infection often exceed in numbers virus replicating infected cells. Why then do antiviral CD8 T cells not do a better job of controlling infection? Although viral strategies for immune evasion are well known, this review will focus on changes in the CD8 T cell that interfere with cytolytic function. Most antiviral CD8 T cells in chronic infection do not express perforin, a molecule that is required for cytolysis. IL-2 and other costimulatory signals can restore cytotoxicity that has been impaired, suggesting a role for CD4 T cell anergy. The chance to eradicate an infection by T cell mediated lysis is undermined after an infection becomes established, in part because the effector immune response is impaired in the setting of chronic antigen. PMID- 12088686 TI - T cell immunity in lymphoid and non-lymphoid tissues. AB - Immune responses to infection or effective vaccination generally result in the development of memory lymphocytes capable of mounting a rapid response to secondary infection. Since most infections initiate in non-lymphoid tissues, defense at these sites may be important for protection. Recent results suggest that a substantial portion of the T cell response to infection is focused in non lymphoid tissues. Furthermore, anatomic localization appears to define phenotypic and functional heterogeneity among antigen-specific memory T cell populations. PMID- 12088685 TI - Neutralizing antibodies against HIV -- back in the major leagues? AB - The past few months have seen encouraging successes for neutralizing antibodies against HIV; human monoclonal antibodies targeting conserved HIV envelope epitopes potently neutralized primary virus isolates, including strains of different clades. In primates, passive immunization with combinations containing human monoclonal antibodies completely prevented infection, even after mucosal virus challenges. Epitopes recognized by the protective monoclonal antibodies are important determinants for protection and provide a rational basis for AIDS vaccine development. PMID- 12088687 TI - NK cell receptors in antiviral immunity. AB - An array of inhibitory and activating receptors initially identified on NK cells are also expressed by conventional CD8+ alphabeta T cells. New evidence strongly implicates these 'NK cell receptors' in modulating NK cell and virus-specific CD8+ T cell responses against a variety of viral infections. Precise regulation of NK cell and T cell responses by these receptors optimizes antiviral immunity while preventing immunological bystander pathology and autoimmunity. PMID- 12088688 TI - Humoral immunity and long-lived plasma cells. AB - A selected fraction of plasmablasts enters the compartment of nondividing, long lived plasma cells to maintain humoral antibody memory. In accord with a current model for lymphocyte homeostasis, the lifetime of long-lived plasma cells is probably regulated by competition for a limited number of survival niches present in splenic red pulp, bone marrow and inflamed tissue. Plasma cells secreting autoantibodies specific for some, but not all, self-antigens are probably 'allowed' to enter the compartment of long-lived plasma cells and provide antibody-mediated 'autoimmune memory' that is resistant to conventional therapies. PMID- 12088689 TI - On differences between immunity and immunological memory. AB - The evolutionary benefits of immunological memory are important: whereas antibodies can be transmitted to offspring by their mother and thereby benefit the species, T cell memory may function to help the individual combat persistent infection in peripheral tissues. Although experimental immunological memory is largely maintained antigen-independently, protective immunity is antigen dependent. PMID- 12088690 TI - Distinct fronto-central N60 and supra-sylvian N70 middle-latency components of the median nerve SEPs as assessed by scalp topographic analysis, dipolar source modelling and depth recordings. AB - OBJECTIVES: To investigate the possible contribution of the second somatosensory (SII) area in the generation of the N60 somatosensory evoked potential (SEP). METHODS: In 7 epileptic patients and in 6 healthy subjects scalp SEPs were recorded by 19 electrodes placed according to the 10-20 system. All epileptic patients but one were also investigated using depth electrodes chronically implanted in the parieto-rolandic opercular cortex. Scalp SEPs underwent brain electrical source analysis. RESULTS: In both epileptic patients and healthy subjects, scalp recordings showed two middle-latency components clearly distinguishable on the basis of latency and scalp distribution: a fronto-central N60 potential contralateral to stimulation and a later bilateral temporal N70 response. SEP dipolar source modelling showed that a contralateral perisylvian dipole was activated in the scalp N70 latency range whereas separate perirolandic and frontal sources were activated at the scalp N60 latency. Depth electrodes recorded a biphasic N60/P90 response in the parieto-rolandic opercular regions contra- and ipsilateral to stimulation. CONCLUSIONS: Two different middle-latency SEP components N60 and N70 can be distinguished by topographic analysis and source modelling of scalp recordings, the sources of which are located in the fronto-central cortex contralateral to stimulation and in the supra-sylvian cortex on both sides, respectively. The source location of the scalp N70 in the SII area is strongly supported by its spatio-temporal similarities with SEPs directly recorded in the supra-sylvian opercular cortex. PMID- 12088691 TI - Movement interference attenuates somatosensory high-frequency oscillations: contribution of local axon collaterals of 3b pyramidal neurons. AB - OBJECTIVES: We examined the effects of movement interference on high-frequency oscillations (HFOs) and N20m in 10 healthy subjects. METHODS: For the movement interference condition, somatosensory evoked magnetic fields (SEFs) following electric median nerve stimulation were recorded during voluntary movement of the digits. For the control condition, the SEFs were recorded without interference. The N20m and HFOs were separated by 3-300Hz and 300-900Hz bandpass filtering. Then, the peak-to-peak amplitudes were measured. RESULTS: Both interference/control amplitude ratios for the N20m and HFOs were smaller than 100%. In contrast, the HFO/N20m amplitude index, which was calculated by dividing the interference/control amplitude ratio for the HFOs with that for the N20m, was significantly greater in the movement interference condition than in the control condition. CONCLUSIONS: Although the overall amplitude of the HFOs was decreased by movement, enhancement of the HFOs by the movement was revealed by the HFO/N20m amplitude index. Thus, we suggest that the HFOs represent activity of the inhibitory interneurons excited by both thalamocortical afferent impulses and excitatory synaptic inputs from pyramidal neurons in area 3b through their local axon collaterals, thereby reflecting both feed-forward and feedback inhibitory effects onto the post-synaptic pyramidal neurons. PMID- 12088692 TI - Intrathalamic non-propagating generators of high-frequency (1000 Hz) somatosensory evoked potential (SEP) bursts recorded subcortically in man. AB - OBJECTIVES: Recently, bursts of high-frequency (1000 Hz) median nerve somatosensory evoked potential (SEP) wavelets were recorded subcortically near and inside the thalamus from deep brain electrodes implanted for tremor therapy. This study aimed to clarify whether these subcortical SEP bursts reflect evoked axonal volleys running in the thalamocortical radiation or a locally restricted intrathalamic response. METHODS: During deep brain electrode implantation, median nerve SEP were recorded in 7 patients sequentially along the subcortical stereotactic trajectory at sites +20 and +10 mm above the respective target nucleus (ventral intermediate thalamus or nucleus subthalamicus). Low- and high frequency SEP components (corner frequency 430 Hz) were analyzed separately with respect to peak latency and amplitude as they changed along the recording trajectory. RESULTS: Individual wavelets of the subcortical 1000 Hz SEP burst showed fixed peak latencies independent from the depth of the electrode penetration; they increased markedly in amplitude with decreasing distance to the thalamus. In contrast, the amplitude gradient between the two recording sites was shallower for the low-frequency SEP component, which peaked earlier at the lower recording site. CONCLUSIONS: Subcortically recorded 1000 Hz SEP wavelet bursts predominantly reflect locally restricted near-field activity, presumably generated in the somatosensory relay nucleus. In contrast, the variable peak latency of the subcortical low-frequency component could reflect postsynaptic potentials sequentially evoked during passage of the lemniscal afferences curving through the thalamus and contributions from the thalamocortical radiation. PMID- 12088693 TI - High intensity magnetic stimulation over the lumbosacral spine evokes antinociception in rats. AB - OBJECTIVES: High intensity magnetic stimulation (MS) applied over the skin can painlessly depolarize superficial and deep nerves and we aimed to evaluate the effectiveness of MS of spinal nerves in evoking a potent analgesic response. METHODS: The MS was administered to adult male Sprague-Dawley rats using a Cadwell MES-10 high-speed magnetic stimulator. A Peltier device and von Frey fibers were used to determine heat and mechanical nociceptive responses of the rats. RESULTS: A brief (5 min) course of MS over the rat's lumbosacral spine produced a long-lasting (30-40 min) and robust (80-90% maximum possible effect) hindpaw antinociceptive effect to both mechanical and heat stimuli. Spinal cord transected rats had intact hindpaw nociceptive withdrawal responses, but transection eliminated MS evoked antinociception, indicating a critical extrasegmental component in the mechanism of MS antinociceptive action. The opiate receptor antagonist naloxone (5 mg/kg, i.p.) completely blocked MS evoked antinociception, demonstrating an opioidergic mechanism for MS antinociception. The alpha(2) adrenoceptor antagonist atipamezole (5 mg/kg, i.p.) slightly reduced the MS antinociceptive response to heat and had no effect on MS antinociception for mechanical stimuli. CONCLUSIONS: These data indicate that MS can evoke a robust, long-lasting antinociceptive effect, which requires an intact supraspinal pathway and is opioidergic mediated. PMID- 12088694 TI - Cortical potentials related to assessment of pain intensity with visual analogue scale (VAS). AB - OBJECTIVES: To elucidate brain mechanisms underlying the psychophysical processes to measure pain intensity, pain-related somatosensory evoked potentials (pain SEPs) following painful CO(2) laser stimulation were studied while employing a task to measure intensity of pain on a visual analogue scale (VAS). METHODS: In 12 healthy subjects, 3 kinds of CO(2) laser stimuli, different in intensity as determined by irradiation duration of 40, 60 and 80ms, were randomly delivered to the left hand dorsum at an irregular interval of 4-6s. The subject was requested to assess the intensity of each pain stimulus and point to the VAS scale by moving a pointer held with the right hand according to the subjective feeling of pain sensation (pain intensity assessment (PIA) condition). For the control condition, the subject moved the pointer to the midpoint of the VAS line irrespective of the pain intensity (control motor task condition). Electroencephalograms were recorded from 21 scalp electrodes, referenced to the linked earlobes, and were averaged time-locked to the stimulus onset for each stimulus duration as well as for each task condition. RESULTS: The VAS scores were 2.8+/-0.5/10 for the stimulus of 40ms duration, 4.8+/-0.8/10 for 60ms and 6.1+/-0.9/10 for 80ms, and showed a highly significant positive correlation with the stimulus duration. Following the early components of pain SEPs which were affected by stimulus duration but not modulated by task conditions, a surface positive peak at latency of 612-642ms was identified exclusively under the PIA condition regardless of the stimulus intensity and was called 'intensity assessment-related potential (IAP)'. The IAP was maximal at the midline parietal area and symmetrically distributed over the scalp. Neither latency nor amplitude of the IAP was significantly different among the 3 different stimulus intensities. CONCLUSIONS: IAP is an event-related potential (ERP) associated with assessment of pain intensity but not influenced by pain intensity itself. From its scalp distribution, it can be assumed that the assessment of pain intensity involves multiple areas in both hemispheres. PMID- 12088695 TI - Diazepam-enhanced beta activity in Sturge Weber syndrome: its diagnostic significance in comparison with MRI. AB - OBJECTIVES: This study was performed to assess the extent of functional involvement of the affected hemisphere in Sturge Weber syndrome in comparison with the uninvolved hemisphere. To this end beta activity in the electroencephalogram (EEG) was measured, both before and after administration of diazepam intravenously (i.v.). METHODS: In 9 patients asymmetry in beta band activity was studied before and after diazepam administration. Several beta bands and asymmetry parameters were calculated. beta band asymmetries were compared with structural abnormalities (magnetic resonance imaging, MRI). RESULTS: Total beta activity was reduced in the involved hemisphere in all patients after diazepam administration. In 3 patients functional abnormalities were found in brain regions that were structurally intact. CONCLUSIONS: Decreased diazepam enhanced beta activity in the EEG is a sensitive criterion of functional abnormality. In patients with subtle structural abnormalities diazepam-enhanced EEG may have added value in diagnosing functional involvement and in monitoring disease progression in patients. PMID- 12088696 TI - PLEDs in Creutzfeldt-Jakob disease following a cadaveric dural graft. AB - INTRODUCTION: Periodic synchronous discharges (PSDs) are a well-known electroencephalographic finding associated with Creutzfeldt-Jakob disease (CJD), but only a few reports have documented the appearance of periodic lateralized epileptiform discharges (PLEDs) in CJD and there has been no discussion as to why PLEDs appear with unilateral (right or left) dominance. CASE REPORT AND DISCUSSION: We report on a 61-year-old man who received a cadaveric dura mater graft and developed CJD 14 years later. Periodic lateralized epileptiform discharges (PLEDs) were observed predominantly in the right hemisphere, coinciding with the location of the dural graft, the presumed source of the CJD agent and PLEDs seen in this case finally developed into PSDs. A similar case has not been reported in the literature and we believe this case serves to further the understanding of the pathophysiology of CJD. PMID- 12088697 TI - EEG evidence for a new conceptualisation of attention deficit hyperactivity disorder. AB - OBJECTIVES: This study investigated the presence of electroencephalographic (EEG) clusters within a sample of children with the inattentive type of attention deficit/hyperactivity disorder (ADHD). METHODS: Subjects consisted of 100 boys with ADHD and 40 age-matched controls. EEG was recorded from 21 sites during an eyes-closed resting condition and Fourier transformed to provide estimates for total power, and relative power in the delta, theta, alpha and beta bands. Factor analysis was used to group sites into 3 regions; frontal, central and posterior. These data were subjected to cluster analysis. RESULTS: Two distinct EEG clusters of children with the inattentive type of ADHD were found. These were characterised by (a) increased high-amplitude theta with deficiencies of delta and beta activities, and (b) increased slow wave and deficiencies of fast wave activity. CONCLUSIONS: These two subtypes are independent of current diagnostic categories, and consist of a cortically hypoaroused group and a group typified by a maturational lag in central nervous system (CNS) development. These results support a re-conceptualisation of ADHD based on the CNS abnormality underlying the disorder rather than the behavioural profile of the child. This has the potential to add a level of predictive validity, which is currently lacking in the present diagnostic systems. PMID- 12088698 TI - EEG harmonic responses to photic stimulation in normal aging and Alzheimer's disease: differences in interhemispheric coherence. AB - OBJECTIVES: The distinction between electroencephalogram (EEG) driving responses in normal aging and Alzheimer's disease (AD) is unclear. This study was conducted to explore the structure of harmonic responses of EEG coherence during photic stimulation (PS) in AD. METHODS: EEG interhemispheric coherence was analyzed for each harmonic responses during PS (white flickers at 5, 10 and 15 flashes/sec) in younger subjects (20-25 years), non-demented elderly subjects (50-67 years) and AD patients (43-66 years). RESULTS: The elderly subjects had higher interhemispheric coherence during PS than the younger subjects, with significant differences found in the 6th (30Hz) and 8th (40Hz) harmonic responses to 5Hz PS. The AD patients had significantly smaller coherence than the elderly subjects in the 2nd (10Hz) and 4th (20Hz) harmonic responses to 5Hz PS, and in the fundamental (10Hz) and 2nd (20Hz) harmonic responses to 10Hz PS. These coherence values did not correlate with duration of the disease or with cognitive functioning as evaluated by MMSE score. CONCLUSIONS: A failure of stimulation related brain activation in AD patients was observed in alpha-related frequencies, suggesting that AD is not the end result of normal aging of the brain, at least in terms of the mechanisms which are involved in PS-related functional organization. PMID- 12088699 TI - A new EEG method for estimating cortical neuronal impairment that is sensitive to early stage Alzheimer's disease. AB - OBJECTIVES: To test the hypothesis that elecetroencephalographic (EEG) analysis is sensitive to cortical neuronal impairment in early stage Alzheimer's disease (AD), and that this analysis correlates with corresponding changes in cerebral blood flow. METHODS: We examined an EEG measure of neuronal impairment in the cerebral cortex in terms of its ability to detect very mild AD. This measure, the mean value of the resting state EEG alpha dipolarity (D(alpha)), approaches unity without cortical sulcal lesions, whereas brains with randomly distributed cortical sulcal lesions lower D(alpha) values well below unity. D(alpha) was evaluated in 25 patients with very mild AD, 33 patients with moderately severe AD, and 56 normal age-matched subjects. These subjects also received SPECT, and strong correlation between D(alpha) and regional cerebral blood flow (rCBF) was observed. RESULTS: D(alpha) values greater than 0.977 correctly classified normal subjects, but also included 10% of very mild AD. D(alpha) values less than 0.952 correctly classified very mild AD as well as moderately severe AD, but also included 10% of normal subjects. D(alpha) values also correlated positively with bilateral temporal-parietal rCBF (a characteristic finding in AD patients); both declined with increasing dementia severity. CONCLUSIONS: Analysis of D(alpha) in this sample supports the hypothesis that early stages of AD can be discriminated from normal aging using measures of cortical neuronal impairment. Furthermore, dementia severity, as reflected by the degree of impairment, is reflected in declining D(alpha) values and increasing variance (greater spread of the D(alpha) values). PMID- 12088700 TI - EEG synchronization upon reward in man. AB - OBJECTIVES: It was tested whether reward in humans is associated with EEG synchronization similar to that seen in animals. METHODS: In two experiments (I and II) the EEG was recorded from frontal, central, and parietal positions before, during, and after drinking or oral stimulation. In Experiment I, subjects (n=11) who had either been thirsty for 16h or had quenched thirst before recordings, drank 400ml of water. In Experiment II, thirsty subjects (n=11) either drank 400ml of water or sucked on a soother. The recording epochs included a 3min baseline, an interval of about 5min during which subjects drank or sucked on the soother, and a 7min post-drinking interval. RESULTS: During the drinking epoch, beta band-power (12-30Hz) was enhanced in both conditions of Experiment I and II, respectively. In Experiment I, after drinking, lower alpha power (8-10Hz) was higher when subjects were thirsty than when they were not. Lower alpha was also enhanced in the post-drinking interval of both conditions of Experiment II, and after sucking, lower alpha synchronization was in addition accompanied by increased theta activity (4-8Hz). CONCLUSIONS: Increased beta activity during drinking and sucking in thirsty subjects presumably reflects non-specific activation related to the motivational strength of sensorimotor regulation during consumatory behavior. The thirst dependent lower alpha synchronization after drinking, generated not only by water consumption but also by surrogate oral stimuli, can be considered a reflection of the drive reducing and rewarding qualities of oral stimulation and consumatory behavior. PMID- 12088701 TI - Effect of amyotrophic lateral sclerosis serum on calcium channels related to spontaneous acetylcholine release. AB - OBJECTIVES: The aim of this work was to further investigate the effect of sera from sporadic amyotrophic lateral sclerosis (ALS) patients on miniature end-plate potentials (MEPP) frequency, by the mouse passive transfer model, and to study whether the transferred serum induces any change in the sensitivity of the L-type voltage-dependent calcium channels (VDCC) to its specific blocker Nitrendipine. METHODS: A total of 35 CF1 mice were divided into 3 groups: (a) ALS group receiving sera from 15 patients that had been clinically and electromyographycally diagnosed as having sporadic ALS; (b) normal group receiving sera from 13 healthy volunteers and from 3 disease control patients, and (c) control group, which was kept untreated. Animals in groups (a) and (b) received daily intraperitoneal injection of 0.5-1ml of serum for 3 days, and 24h later the left hemidiaphragm was excised for electrophysiological recordings. RESULTS: Analysis of MEPPs frequency recorded from ALS group showed that 3 of them induced an increase in spontaneous neurotransmitter release while in 4 a decrease was observed, suggesting that sera alter spontaneous secretion as result of an increased or decreased Ca(2+) influx through the normally involved N-type or L-type VDCC, respectively. When the effect of Nitrendipine, an L-type VDCC blocker, was studied on ALS sera-injected mice, we found variable responses to the drug: only two mice showed control sensitivity to Nitrendipine, while in 7 its action was lower and surprisingly in 4 was greater than that without the drug. CONCLUSIONS: These results suggest that ALS sera contain factor(s) that are able to modify spontaneous neurotransmitter release by altering calcium current through L-type and N-type VDCC, and even inducing changes in the sensitivity to the L-type VDCC blocker. PMID- 12088702 TI - The cause of slowed forearm median conduction velocity in carpal tunnel syndrome: a Palmar stimulation study. AB - OBJECTIVES: To elucidate the etiopathogenesis of decreased forearm median motor conduction velocity (FMMCV) in carpal tunnel syndrome (CTS), we used segmental stimulation at the palm, wrist and antecubital fossa to determine conduction block at wrist and calculate and compare the segmental median motor conduction velocity (MMCV) to determine the pathogenesis. BACKGROUND: The cause of the decreased FMMCV in CTS remains unclear. Animal models have supported retrograde axonal atrophy as the cause. Some authors believe standard FMMCV, calculated by subtracting the distal latency, may not represent an exact assessment of FMMCV but rather the velocity of small fibers that persist throughout the carpal tunnel. SUBJECTS AND METHODS: Patients with clinical symptoms and signs of CTS which had been confirmed with standard electrodiagnosis, were included. The patients were divided into two groups: one with reduced FMMCV <50m/s (Group I, n=20) and the other with normal FMMCV>50m/s (Group II, n=40). Age-matched volunteers served as controls (n=60). We used palm, wrist and antecubital stimulation, and recorded compound muscle action potential (CMAP) amplitudes at the abductor pollicis brevis (APB) muscle. Based on a ratio of the CMAP amplitudes obtained from wrist and palm stimulation (W/P ratio) and the latency differences, we calculated the W/P ratio and the across wrist MMCV (AWMMCV) and FMMCV and compared and correlated them between two patient groups. RESULTS: There was no difference in median motor and sensory distal latency between Groups I and II. CMAP and sensory nerve action potential amplitudes were reduced in Group I compared with Group II, but the difference was only marginally significant. Four patients had a significant reduction of the W/P ratio in Group I, compared with 7 patients in Group II, which did not reach a significance. Sixteen patients (80%) in Group I demonstrated no conduction block. Furthermore, Group I showed significantly decreased FMMCV when compared with Group II; however, AWMMCV was not significantly reduced in Group I, suggesting that decreased FMMCV does not result from a decrease in AWMMCV. CONCLUSIONS: There was no significant motor conduction block and no correlation of the FMMCV and AWMMCV in CTS patients with a decrease of FMMCV, suggesting retrograde axonal atrophy, and not selective conduction block of the large fibers at the wrist, is the direct cause of decreased FMMCV in CTS. PMID- 12088703 TI - Multi-MUP analysis of laryngeal muscles. AB - We performed quantitative motor unit potential (MUP) analysis of the thyroarytenoid (TA) and cricothyroid (CT) muscles, using multi-MUP-analysis in 40 healthy volunteers. The method is well tolerated, easy to perform, and examination of one muscle takes 5-10min. The mean MUP amplitude of both muscles was significantly larger in men than in women. The method can safely be used in clinical routine. PMID- 12088705 TI - Post-exercise facilitation and depression of M wave and motor evoked potentials in healthy subjects. AB - OBJECTIVES: To characterize so-called central fatigue, the effect of various levels of exercise on central and peripheral motor potentials were compared. METHODS: Thirteen healthy subjects performed 4 levels of exercise following isometric dorsiflexion of the foot. Post-exercise recordings from the anterior tibial muscle of motor evoked potentials (MEP) evoked by transcranial magnetic stimulation (TMS) and M wave evoked by electrical stimulation of the peroneal nerve were performed. RESULTS: After 5 s the post-exercise MEP amplitude increased. The increase was related to the degree of work performed. Subsequently, there was a gradually decrease of amplitude reaching statistical significance after 15 min. The area of the M wave increased significantly after 10 s and returned to baseline after 2-3 min. CONCLUSIONS: Facilitation and depression of MEP after fatiguing exercise is at least partly a peripheral phenomenon dependent on the level of exercise performed. PMID- 12088704 TI - Combined monitoring of motor and somatosensory evoked potentials in orthopaedic spinal surgery. AB - OBJECTIVES: To demonstrate possible advantages of combined (motor and sensory) versus single modality (either motor or sensory) intra-operative spinal cord monitoring and to investigate risk factors for post-operative neurological sequelae. METHODS: Recordings of lower limb motor evoked potentials (MEPs) to multi-pulse transcranial electrical stimulation (TES), and tibial nerve somatosensory evoked potentials (SEPs), were attempted during 126 operations in 97 patients (79 with spinal deformity and 18 with miscellaneous spinal disorders). RESULTS: Combined motor and sensory monitoring was successfully achieved in 104 of 126 (82%) operations. No response to either modality could be recorded in two patients with Friedreich' s ataxia. In 18 patients monitoring was possible in only one modality: SEPs could not be recorded in two patients and MEPs in 16. Significant intra-operative EP changes occurred in one or both modalities in 16 patients; in association with instrumentation in 10 cases, and with systemic changes in 6. After appropriate remedial measures, SEPs recovered either fully or partially in 8/8 patients and MEPs in only 67% (10/15 patients). New deficits were present post-operatively in 6 of the 16 patients with abnormal intra-operative EPs. Normal MEPs at the end of the operation correctly predicted the absence of new motor deficits in all cases. SEPs either remained unchanged or recovered fully after remedial measures in 3 patients with new post-operative motor deficits. Neurological complications were more frequent in patients with miscellaneous spinal disorders and/or pre-existing neurological deficits. No complications occurred in patients with idiopathic scoliosis. CONCLUSIONS: Combined SEPs and multi-pulse TES-MEPs provide a safe, reliable and sensitive method of monitoring spinal cord function in orthopaedic surgery. This method is superior to single modality techniques, both for increasing the number of patients in whom satisfactory monitoring of spinal cord function can be achieved and, for improving the sensitivity and predictivity of monitoring. Combined SEP/MEP methods may enhance the impact of neuromonitoring on the intra-operative management of the patient and favourably influence neurological outcome. PMID- 12088706 TI - Early and late inhibition in the human motor cortex studied by paired stimulation through subdural electrodes. AB - OBJECTIVES: To evaluate the focal nature of the early and late inhibition of corticospinal neurons demonstrated by a paired-pulse stimulation paradigm. METHODS: We performed paired-electric pulse stimulation studies using subdural electrodes implanted in 4 patients with intractable partial epilepsy. RESULTS: Inhibition of motor evoked potentials in the first dorsal interosseous muscle was obtained by paired-pulse stimulation of the hand motor cortex (M1) with a subthreshold conditioning stimulus at conditioning-test intervals between 1 and 6ms. This early inhibition was abolished when the conditioning stimulus was moved to the sensory cortex (S1) or the arm M1. The inhibition was also produced by paired-pulse stimulation of the hand M1 with a suprathreshold conditioning stimulus between 50 and 300ms in all 3 patients. This late inhibition was still recognized when moving the conditioning stimulus to the hand S1 only in one of 3 patients. CONCLUSIONS: The early inhibition arises from very small areas in the M1 and is little mediated by neuronal circuits in the S1. On the other hand, the focal nature of the late inhibition is complicated and it arises mainly from the M1 but the S1 may be related to the generation of the late inhibition in some cases. PMID- 12088707 TI - Influence of internal globus pallidus stimulation on motor cortex activation pattern in Parkinson's disease. AB - OBJECTIVES: Indications of the functional neurosurgical treatments become more and more numerous, however, few methods were used to study the mechanism of action and some discrepancies came to light. We assessed the influence of internal globus pallidus (GPI) stimulation and L-Dopa on cortical activation during the preparation and execution phases of the movement compared to clinical improvement of Parkinson's disease. METHODS: We recorded the movement-related cortical potential and movement-related desynchronization. RESULTS: The Unified Parkinson's Disease Rating Scale was improved by 46% under stimulation and 64% under stimulation with L-Dopa. Premovement desynchronization was significantly increased on central contralateral derivation under stimulation with L-Dopa and decreased on frontocentral ipsilateral derivation under stimulation with and without L-Dopa. Movement desynchronization was improved on the contralateral motor cortex under stimulation with and without L-Dopa (benefit correlated with bradykinesia improvement). The movement-related postmotor potential was significantly increased under stimulation with L-Dopa. CONCLUSIONS: GPI stimulation alone influenced the premotor cortex activation during the planning and induced a selective and focal effect on the organisation of motor cortical activity during the movement execution which may explain bradykinesia improvement. The motor cortex activation improvement under stimulation with L Dopa concerned both the movement preparation and execution but remained very localised to the contralateral motor cortex. PMID- 12088708 TI - The correlation between muscle and nerve fiber conduction velocities in thenar muscle is lost in case of carpal tunnel syndrome. AB - OBJECTIVE AND METHODS: This study investigated the relationship between muscle fiber conduction velocity (MFCV) and motor nerve conduction velocity (MNCV) in thenar muscle of 20 normal subjects and of 20 patients suffering from a moderate carpal tunnel syndrome. Our goal was to confirm the positive correlation between MFCV and MNCV and to assess the influence of carpal tunnel syndrome on this relationship. MFCV was calculated in voluntarily contracted thenar muscle using a multi-channel surface recording and a spike-triggered averaging technique. RESULTS: MFCV values ranged between 2.6 and 7.2 m/s (mean+/-SEM: 4.5+/-0.3) in normal subjects and between 3.5 and 6.9 m/s (4.7+/-0.2) in patients. Subjects and patients did not differ regarding MFCV values, but a correlation between MFCV and MNCV was found in normal subjects (P=0.0005) and not in patients (P=0.54). CONCLUSIONS: A correlation between muscle and nerve conduction velocities existed in healthy subjects but was lost in case of moderate carpal tunnel syndrome. MFCV appeared to be insensitive to focal nerve conduction slowing. PMID- 12088709 TI - Movement-related EEG potentials are force or end-effector dependent: evidence from a multi-finger experiment. AB - OBJECTIVES: This study examined behavioral and electrocortical responses in producing 3 levels of force (25, 50 and 75% of MVC) at a constant rate of force development with each of 4 fingers both during the achievement of the desired force (ramp phase) and its maintenance (static phase). We were particularly interested in describing in more detail the interaction between nominal force and finger on various components of movement-related potential (MRP) associated with preparation and execution of isometric force production tasks. METHODS: Our experimental design systematically controlled the rate of force development while nominal force level was experimentally manipulated during isometric force production tasks. We applied time-domain averaging of EEG single trials in order to extract 3 components of MRP (BP(-600 to -500); MP(-100 to 0); MMP) preceding and accompanying behavioral responses. RESULTS: Overall, as in our previous research the effect of force per se was not reflected in the EEG components. However, we did find an interaction between finger and force level in both the Bereitshaftspotential (BP) and motor potential (MP) components of the movement related potentials. While the middle, ring and little finger produced no differences in EEG components at any of the 3 force levels, the index finger did. We further correlated the force trajectory and the EEG time series with the highest correlations found in the lowest force level with the index finger. As the force level was increased, the correlation was significantly reduced. CONCLUSIONS: Overall, the whole complex of MRP components and evolution of EEG time series during multi-finger isometric force production tasks reflect a combination of factors including the primary end-effector performing the task and interaction of end-effector and the amount of nominal force. PMID- 12088710 TI - Intensity-dependent effects of 1 Hz rTMS on human corticospinal excitability. AB - OBJECTIVES: This study explored whether the effects of repetitive transcranial magnetic stimulation (rTMS) on corticospinal excitability are dependent on the stimulation intensity and examined the effect of rTMS on inhibitory function. METHODS: Nine normal volunteers received 15min of 1Hz rTMS at 85 and 115% of the resting motor threshold (RMT). Cortical excitability was measured before and after rTMS. RESULTS: rTMS at both intensities produced an increase in the RMT but only 115% stimulation reduced the size of motor evoked potentials (MEPs). rTMS had no effects on the cortical silent period or cortical inhibition measured with paired pulse TMS. CONCLUSIONS: The effects of 1Hz rTMS on motor cortex excitability are partially dependent on stimulus intensity and the effects of rTMS on motor thresholds and MEP size may differ. PMID- 12088711 TI - Multi-channel EMG of the M. triceps brachii in rats during treadmill locomotion. AB - OBJECTIVES: The study aims at a precise characterisation of intramuscularly varying recruitment patterns within the triceps brachii muscle (long and lateral head; proximal, medial, distal regions) in the time course of averaged step cycles during locomotion. METHODS: The triceps brachii muscle of 15 Hannover rats was investigated with a supramuscular 16-electrodes grid during treadmill locomotion. Multi-channel electromyogram (EMG) was recorded simultaneously with high-speed videography. The rectified and smoothed EMG was time-normalised. EMG profiles and dynamic EMG-map series were calculated. Differences between EMG distribution patterns were tested by multivariate analysis of variance. RESULTS: In the pre-stance phase EMG activity increased especially in the proximal long head. It most likely propagated from lower muscle layers of the long head. During stance phase the EMG activity of the lateral head rose steeply and exceeded those of the long head in short time. The fastest steps show the highest EMG amplitudes. CONCLUSIONS: EMG registrations with grid electrodes help in the identification of intramuscular co-ordination processes during locomotion. While the EMG profiles characterise the time course, the topographical distribution is better represented in dynamic EMG interference maps. The dynamic changing activation patterns of triceps brachii depend on the phase of the step cycle. This clearly indicates the different functions of the muscle heads. PMID- 12088712 TI - Thalamocortical dysrhythmia and the thalamic reticular nucleus in behaving rats. AB - OBJECTIVES: The aim of this study was to investigate the effects of bilateral chemical lesion of the rostral pole of the thalamic reticular nucleus on EEG activities in freely moving rats applying quantitative analysis and brain mapping of power spectra distribution. METHODS: Ketamine-sedated Sprague-Dawley rats were implanted to monitor behavioral states with frontoparietal electrodes in a first series of experiments and with multiple electrodes along the antero-posterior axis (F1, F2, F7, F8, T3, T4, P3, P4) in a second series. Monopolar and bipolar recordings were obtained in animals stereotaxically injected with ibotenic acid into both rostral poles of the thalamic reticular nucleus. Long-term video-EEG recordings and brain mapping based on quantitative spectral analysis were made. RESULTS: Two forms of dysrhythmia gradually emerged in the neocortical EEG at 12 24h post-injection: potentiation of theta waves and spontaneous high-voltage spindles (HVS) at 4.5-8Hz frequency. Brain mapping during these dysrhythmia shows highest power posteriorly (parietotemporal) for theta and mesiofrontally for HVS. CONCLUSIONS: Given the lack of inhibitory intrinsic interneurons in the rat thalamus, bilateral destruction of a small part of the solely GABAergic population may promote cortical dysrhythmia (probably by dis-inhibition). The topographic differences in power might indicate different involved structures. PMID- 12088713 TI - Variation in the response to transcranial magnetic brain stimulation in the general population. AB - OBJECTIVES: The aim of this study is to describe the variability and other characteristics of the motor evoked potential (MEP) to transcranial magnetic stimulation (TMS) in a large database. METHODS: One hundred fifty one subjects, including 17 sib pairs, free of neurological or psychiatric disease and on no neuroactive medications were studied with uniform techniques. Nineteen were studied on 3 occasions. Measures included MEP threshold (N=141) during rest and voluntary muscle activation and the response to paired TMS (subthreshold conditioning stimulus) at interstimulus intervals (ISIs) of 3, 4, 10, and 15ms (N=53). RESULTS: There was a large variability in all the measures. Approximately 40-50% of this appeared to come from within-subjects variation or experimental error. The MEP threshold data were skewed downward, but normalized with log transformation. The paired-pulse ratios (conditioned/unconditioned MEP) were normally distributed except those from the 3ms ISI which had no lower tail and could not be normalized. There were subjects showing inhibition and others showing facilitation at all ISIs. There were no correlations in any of the data with age or sex, but MEP thresholds were highly correlated within sibs. CONCLUSIONS: These data should be useful for planning, analyzing, and interpreting TMS studies in healthy and patient populations. PMID- 12088714 TI - Inhibition, response mode, and stimulus probability: a comparative event-related potential study. AB - OBJECTIVES: In the present study, effects of response mode (finger movement or counting) and stimulus probability on inhibitory processes were studied. METHODS: Electroencephalographic activity was registered in a visual go/nogo paradigm. Subjects either responded manually to go stimuli or counted silently the occurrence of each go stimulus in different conditions. In both response mode conditions, response probability was varied. RESULTS: For finger movement and counting, similar N2 and P3 go/nogo effects were found. The amplitude of the nogo N2 and nogo P3 varied as a negative function of nogo stimulus probability. The go P3 varied as a negative function of go stimulus probability. In the manual condition, however, the descending flank of the go N2 at anterior electrode sites was more negative in the 0.50go and 0.75go probability trials than in the 0.25go probability trials. CONCLUSIONS: The results of the present study confirm the hypothesis that differences between go and nogo event-related potentials are not dependent on overt movement-related potentials. It could be speculated that the probability effect on the N2 amplitude in go trials in the manual condition has to be explained in terms of a modulation of the strength of motoric preparation processes varying as a positive function of the probability of the go stimulus. PMID- 12088716 TI - Hormesis: changing view of the dose-response, a personal account of the history and current status. AB - This paper provides a personal account of the history of the hormesis concept, and of the role of the dose response in toxicology and pharmacology. A careful evaluation of the toxicology and pharmacology literatures suggests that the biphasic dose response that characterizes hormesis may be much more widespread than is commonly recognized, and may come to rival our currently favored ideas about toxicological dose responses confined to the linear and threshold representations used in risk assessment. Although hormesis-like biphasic dose responses were already well-established in chemical and radiation toxicology by the early decades of the 20th century, they were all but expunged from mainstream toxicology in the 1930s. The reasons may be found in a complex set of unrelated problems of which difficulties in replication of low-dose stimulatory responses resulting from poor study designs, greater societal interest in high-dose effects, linking of the concept of hormesis to the practice of homeopathy, and perhaps most crucially a complete lack of strong leadership to advocate its acceptance in the right circles. I believe that if hormesis achieves widespread recognition as a valid and valuable interpretation of dose-response results, we would expect an increase in the breadth of evaluations of the dose-response relationship which could be of great value in hazard and risk assessment as well as in future approaches to drug development and/or chemotherapeutics. PMID- 12088717 TI - Carcinogenicity of the aromatic amines: from structure-activity relationships to mechanisms of action and risk assessment. AB - Aromatic amines represent one of the most important classes of industrial and environmental chemicals: many of them have been reported to be powerful carcinogens and mutagens, and/or hemotoxicants. Their toxicity has been studied also with quantitative structure-activity relationship (QSAR) methods: these studies are potentially suitable for investigating mechanisms of action and for estimating the toxicity of compounds lacking experimental determinations. In this paper, we first summarized the QSAR models for the rodent carcinogenicity of the aromatic amines. The gradation of potency of the carcinogenic amines depended firstly on their hydrophobicity, and secondly on electronic (reactivity, propensity to be metabolically transformed) and steric properties. On the contrary, the difference between carcinogenic and non-carcinogenic aromatic amines depended mainly on electronic and steric properties. These QSARs can be used directly for estimating the carcinogenicity of aromatic amines. A two-step prediction is possible: (1) estimation of yes/no activity; (2) if the answer from step 1 is yes, then prediction of the degree of potency. The QSARs for rodent carcinogenicity were put in a wider context by comparing them with those for: (a) Salmonella mutagenicity; (b) general toxicity; (c) enzymatic reactions; (d) physical-chemical reactions. This comparative QSAR exercise generated a coherent global picture of the action mechanisms of the aromatic amines. The QSARs for carcinogenicity were similar to those for Salmonella mutagenicity, thus pointing to a similar mechanism of action. On the contrary, the general toxicity QSARs (both in vitro and in vivo systems) were mostly based on hydrophobicity, pointing to an aspecific mechanism of action much simpler than that for carcinogenicity and mutagenicity. The oxidation of the amines (first step in the main metabolic pathway leading to carcinogenic and mutagenic species) had identical QSARs in both enzymatic and physical-chemical systems, thus providing evidence for the link between simple chemical reactions and those in biological systems. The results show that it is possible to generate mechanistically and statistically sound QSAR models for rodent carcinogenicity, and indirectly that the rodent bioassay is a reliable source of good quality data. PMID- 12088718 TI - Lycopersicon assays of chemical/radiation genotoxicity for the study of environmental mutagens. AB - From a literature survey, 21 chemicals are tabulated that have been evaluated in 39 assays for their clastogenic effects in Lycopersicon. Nineteen of the 21 chemicals are reported as giving a positive reaction (i.e. causing chromosome aberrations). Of these, five are reported positive with a dose response. In addition, 23 assays have been recorded for six types of radiation, all of which reacted positively. The results of 102 assays with 32 chemicals and seven types of radiation tested for the induction of gene mutations are tabulated, as well as 20 chemicals and/or radiation in combined treatments. The Lycopersicon esculentum (2n=24) assay is a very good plant bioassay for assessing chromosome damage both in mitosis and meiosis and for somatic mutations induced by chemicals and radiations. The Lycopersicon bioassay has been shown to be as sensitive and as specific an assay as other plant genotoxicity assays, such as Hordeum vulgare, Vicia faba, Crepis capillaris, Pisum sativum and Allium cepa and should be considered in further studies in assessing clastogenicity. Tests using L. esculentum can be made for a spectrum of mutant phenotypes of which many are identifiable in young seedlings. PMID- 12088719 TI - Spectrum of styrene-induced DNA adducts: the relationship to other biomarkers and prospects in human biomonitoring. AB - Styrene is an important industrial chemical that has shown genotoxicity in many toxicology assays. This is believed to be related to the DNA-binding properties of styrene-7,8-oxide (SO), a major metabolite of styrene. In this review, we have summarized knowledge on various aspects of styrene genotoxicity, especially in order to understand the formation and removal of primary DNA lesions, and the usefulness of biomarkers for risk assessment. Biological significances of specific DNA adducts and their role in the cascade of genotoxic events are discussed. Links between markers of external and internal exposure are evaluated, as well as metabolic aspects leading to the formation of DNA adducts and influencing biomarkers of biological effect. Finally, we suggest a design of a population study, which may contribute to our understanding genotoxic events in the exposure either to single xenobiotic or complex mixture. PMID- 12088720 TI - In vitro and in vivo characterisation of [11C]-DASB: a probe for in vivo measurements of the serotonin transporter by positron emission tomography. AB - 3-Amino-4-(2-dimethylaminomethyl-phenylsulfanyl)-benzonitrile, labeled with carbon-11 ([11C]-DASB), is a recently introduced radiotracer for imaging the serotonin transporter (SERT) by positron emission tomography (PET). A series of in vitro and in vivo experiments were performed to further characterise the properties of [11C]-DASB as an in vivo imaging agent for SERT. In vitro binding assays confirmed that DASB binds specifically to SERT with nanomolar affinity and high selectivity over a large number of other receptors, ion-channels and enzymes in the central nervous system. Ex vivo, [11C]-DASB binding in rat brain was shown to be saturable (ED(50) of 56 nmoles/kg), and sensitive to both the number of available SERT binding sites and the number of viable serotonin neurons. Estimates of the radiation dose in man were extrapolated from rat biodistribution data (effective dose 5.5 E-03 mSv/MBq; critical organ --urinary bladder wall). Together with previous studies, the present findings indicate that [11C]-DASB is a very useful radiopharmaceutical for probing changes in SERT densities using PET imaging in the living human brain. PMID- 12088721 TI - Synthesis, in vitro and in vivo pharmacology of a C-11 labeled analog of CP 101,606, (+/-)threo-1-(4-hydroxyphenyl)-2-[4-hydroxy-4-(p [11C]methoxyphenyl)piperidino]-1-propanol, as a PET tracer for NR2B subunit containing NMDA receptors. AB - A carbon-11 labeled methoxyl analog of CP-101,606, (+/-)threo-1-(4-hydroxyphenyl) 2-[4-hydroxy-4-(p-[11C]methoxyphenyl)piperidino]-1-propanol [(+/-)[11C]1], was synthesized as a new subtype-selective PET radioligand for NMDA receptors. The in vitro binding studies using rat brain slices demonstrated that (+/-)[11C]1 shows an extremely high-specific binding to the NR2B subunit of NMDA receptors. In contrast to the in vitro binding, the in vivo binding to mouse and monkey brains showed no apparent specific localization of the radioactivity in any of the brain regions. Metabolism and physicochemical properties such as the lipophilicity of (+/-)[11C]1 seemed unlikely to affect the in vivo (+/-)[11C]1 binding. Among the various endogenous ligands acting at the NMDA receptors, polyamines (spermine and spermidine) and divalent cations (Mg(2+,) Zn(2+,) and Ca(2+)) strongly inhibited the in vitro (+/-)[11C]1 binding. Thus, the present studies point to the possibility that the polyamines and cations behave as endogenous inhibitors for (+/-)[11C]1 binding, leading to the loss of the specific binding in vivo. PMID- 12088722 TI - Evaluation of [76Br]FBAU 3',5'-dibenzoate as a lipophilic prodrug for brain imaging. AB - [76Br]FBAU is a potential PET tracer for assessing proliferation. This study proposes that [76Br]FBAU 3',5'-dibenzoate has higher blood-brain-barrier permeability than [76Br]FBAU itself and thus might be better suited for applications in the brain. The brain uptake indexes of the two compounds measured after carotid injection (29.6 +/- 13.9 for [76Br]FBAU 3',5'-dibenzoate, versus 10.0 +/- 8.7 for [76Br]FBAU) support this claim. Biodistribution study also showed that the brain accumulation of activity was higher in rats injected with [76Br]FBAU 3',5'-dibenzoate than with [76Br]FBAU (0.119+/-0.023 DUR at 1 h, versus 0.061 +/- 0.006). [76Br]FBAU 3',5'-dibenzoate was relatively stable in rat plasma, gradually being hydrolyzed to [76Br]FBAU exponentially with a calculated half-life of 0.8 h. DNA incorporation of [76Br]FBAU was also confirmed. The results presented support the hypothesis that the 3',5'-dibenzoate can act as a prodrug for FBAU and deliver more radiolabeled nucleoside to the brain. PMID- 12088723 TI - In vitro and in vivo characteristics of a human colon cancer cell line, SNU-C5N, expressing sodium-iodide symporter. AB - Rat NIS (rNIS) genes were transfected into a human colon cancer cell line (SNU C5) by lipofection. The transfected cells (SNU-C5N) exhibited an increase 125I uptake to a level 10 times higher than the untransfected SNU-C5 cells. The addition of 300 microM DIDS, an anion channel blocker, to the culture media led to a 2.35 times increase of 125I uptake in the cells. For the first 10 minutes, up to 70% of the cellular radioactivity was released into the medium. In the biodistribution study using SNU-C5N-xenografted mice, the %ID/g of the SNU-C5N tumors at 1, 3, 6, and 12 h after the 125I injection were 4.43%, 1.09%, 1.05%, and 0.05%, respectively, which were significantly higher than those for the SNU C5 tumors (P<0.05). In tumor imaging, the SNU-C5N-xenografted tumor was clearly visible. In this study, NIS lipofection is efficient for triggering significant iodide uptake by a nonthyroidal tumor. However, for an increased therapeutic effect, the key issue is iodide retention in the target tissue. PMID- 12088724 TI - Radiosynthesis and mouse brain distribution studies of [11C] CP-126,998: a PET ligand for in vivo study of acetylcholinesterase. AB - The selective, reversible acetylcholinesterase inhibitor 5,7-Dihydro-7-methyl-3- [2-[1-(phenylmethyl]-4-piperidinyl]ethyl]-6H-pyrrolo[3,2-f]-1,2-benzisoxazol3-6 one (CP-126,998) was labeled with C-11 iodomethane via base-promoted alkylation of the lactam nitrogen. [11C] CP-126,998 was synthesized in good radiochemical yield (13-29% non-decay corrected) and high specific radioactivity (177-418 GBq/micromol). In vivo mouse biodistribution studies reveal [11C] CP-126,998 to localize preferentially in striatal tissue, a region known to be rich in acetylcholinesterase. Competitive blocking studies using a variety of acetylcholinesterase inhibitors (diisopropylfluorophosphate, tacrine, CP-118,954) verified the specificity of the PET radiotracer for brain acetylcholinesterase. PMID- 12088725 TI - In vitro and in vivo evaluation of a 99mTc(I)-labeled bombesin analogue for imaging of gastrin releasing peptide receptor-positive tumors. AB - A new radiolabeled bombesin analogue, [99mTc(I)-PADA-AVA]bombesin (7-14), was synthesized and in vitro and in vivo characterized. High affinity and rapid internalization were obtained in binding assays. A specific binding towards gastrin releasing peptide receptors-positive tissues, pancreas and tumor, was observed in CD-1 nu/nu mice bearing PC-3 prostate adenocarcinoma xenografts. We therefore conclude that [99mTc(I)-PADA-AVA]bombesin (7-14) might have promising characteristics for applications in nuclear medicine, namely for diagnosis of GRP receptor overexpressing tumors. PMID- 12088726 TI - Dual SPECT of dopamine system using [99mTc]TRODAT-1 and [123I]IBZM in normal and 6-OHDA-lesioned formosan rock monkeys. AB - Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by a severe loss of the dopaminergic neurons in the substantia nigra pars compacta. In this study, we evaluated pre- and post-synaptic binding sites of the dopamine system in three normal and one parkinsonian monkeys using simultaneous [99mTc]TRODAT-1 and [123I]IBZM imaging. The parkinsonian monkey was induced by injecting 6-hydroxydopamine (6-OHDA) bilaterally into the medial forebrain bundle under MRI guidance. [99mTc]TRODAT-1 (targeting dopamine transporters) and [123I]IBZM (targeting D(2)/D(3) receptors) were administered almost simultaneously and the SPECT images were acquired over 4 h using a dual headed gamma camera equipped with ultra-high resolution fan-beam collimators. Data were obtained using energy window of 15% centered on 140 keV for 99mTc in conjunction with 10% asymmetric energy window in a lower bound at 159 keV for 123I. Single SPECT studies of [99mTc]TRODAT-1 and [123I]IBZM were also performed. We found a comparable image quality and uptake ratios between single- and dual isotope studies. There are higher TRODAT-1 uptakes in the control monkeys than the 6-OHDA-lesioned monkey. The uptake of [123I] IBZM showed no significant difference between controls and 6-OHDA-lesioned monkey. Our results suggest that dual isotope imaging using [99mTc]TRODAT-1 and [123I]IBZM may be a useful means in evaluating the changes of both pre- and post-synaptic dopamine system in a primate model of parkinsonism. PMID- 12088727 TI - Synthesis and evaluation of 99mTc(CO)(3)-DTPA-folate as a folate-receptor targeted radiopharmaceutical. AB - A folate-receptor-targeted 99mTc-radiopharmaceutical, [99mTc]Tc(CO)(3)DTPA folate, was prepared by heating [99mTc]Tc(CO)(3)(H(2)O)(3)(+) in an aqueous solution of the previously reported DTPA-folate conjugate. The radiotracer was HPLC purified (> 98% radiochemical purity) and evaluated in vitro and in vivo as an agent for targeting folate-receptor-positive cells. [99mTc]Tc(CO)(3)DTPA folate experienced high, folate-receptor-specific uptake in human KB tumor cells. Intravenous administration of [99mTc]Tc(CO)(3)DTPA-folate to athymic mice bearing KB cell tumor xenografts resulted in 99mTc tumor uptake of 1.8 +/- 0.5 and 3.3 +/ 0.2%ID/g (n = 3) at 30 minutes and 4 hours post-injection, respectively. Tumor uptake was reduced when folic acid was co-administered with the intravenous [99mTc]Tc(CO)(3)DTPA-folate, consistent with radiopharmaceutical localization being mediated by the folate receptor. PMID- 12088728 TI - Pharmacokinetics and brain distribution in non human primate of R(-)[123I]DOI, A 5HT(2A/2C) serotonin agonist. AB - Our goal was to synthesize with high specific activity R(-)-1-(2,5-Dimethoxy-4 [123I]iodophenyl)-2-aminopropane [R(-)[123I]DOI], an in vitro potent and selective 5-HT(2A/2C) serotonin agonist, and study in vivo its plasma pharmacokinetics and brain distribution in baboon by SPECT. The purpose was to evaluate this radiotracer as a potential tool in discerning the role of the agonist high affinity state of 5-HT(2) receptors in depression and other neurological disorders. The radiotracer was prepared by electrophilic radioiodination of the N-trifluoroacetyl precursor of R(-)-1-(2,5 Dimethoxyphenyl)-2-aminopropane [R(-)DMA-TFA] with high-purity sodium [123I]iodide in the presence of chloramine-T, followed by amino deprotection with KOH in isopropanol (labeling yield: 73%, radiochemical yield: 62%, radiochemical purity: 99%). In vivo studies in baboon showed high accumulation of radioactivity in thalamus, the frontoparietal cortex, temporal, occipital and the striatum regions, with slightly lower accumulation in the midbrain and cerebellum. Ketanserin did not displaced the radioactivity in any of these brain regions. Plasma metabolite analysis was performed using methanol protein precipitation, the methanol fractions contained from 68% to 92% of the mixture of a labeled metabolite and parent compound. The recovery coefficient of unmetabolized R( )[123I]DOI was 68%. The percent parent compound present in the extracted fraction, measured by HPLC, decreased gradually with time from 99.8% to 0.3% still present after 4.7 hours post injection whereas the percentage of the only one detected metabolite increased conversely. Free fraction determination (f(1)), was 31 +/- 0.9% (n = 3). For comparison purposes, ex-vivo brain distribution, displacement and metabolite analysis was also carried out in rodents. Although R( )[123I]DOI displayed good brain uptake and localized in serotonergic areas of the brain, its target to non target ratio and its insensitivity to ketanserin displacement suggest high nonspecific uptake, therefore non potentially useful as brain imaging radiotracer for visualization of the agonist high-affinity state of 5-HT(2A) receptors and for visualizing 5-HT(2C) receptors by SPECT. PMID- 12088729 TI - Preparation and preliminary evaluation of 99mTc-EC-For-MLFK. AB - For-Met-Leu-Phe-Lys (For-MLFK), a chemotactic peptide that binds with high affinity to granulocytes and monocytes, was labeled with 99mTc using ethylene dicysteine (EC) as the metal chelating system. EC was selected because of the rapid renal excretion of its 99mTc-complex and therefore, was expected to enhance the degree of urinary elimination of the peptide-conjugate. 99mTc-EC-For-MLFK was prepared using a preformed chelate approach. After incubation of 99mTc-EC-For MLFK with total blood, 68.1% of the labeled peptide was associated with WBC and 86% of this cell-associated activity was bound to granulocytes. Biodistribution studies in normal mice revealed a very fast blood clearance (4.1% and 0.6% of I.D. in blood at respectively 5 and 60 min p.i.). However, elimination of the labeled peptide proceeds mainly via the hepatobiliary system (24.5% of I.D. in liver and 48.8% of I.D. in intestines at 60 min p.i.) and to a much lower degree via the kidneys (17.9% in renal system at 60 min p.i.). From these results, it is concluded that 99mTc-EC-For-MLFK is not suited to image infections, despite its high binding to granulocytes, since it leads to high, non-specific, abdominal activity. PMID- 12088730 TI - Experimental radiation synovectomy in rabbit knee with holmium-166 ferric hydroxide macroaggregate. AB - Holmium-166 ferric hydroxide macroaggregate (Ho-166 FHMA) particles possess two important properties for radiosynovectomy; relatively short half-life of the radioisotope and appropriate carrier size. Both these minimize radioactive leakage from the treated joint. This study was conducted to assess the effects of Ho-166 FHMA on synovium and synovial fluid in rabbit knee joints. Whole-knee autoradiography was utilized to determine distribution of radioactivity after intra-articular Ho-166 FHMA injection. Intra-articular injection of Ho-166 FHMA resulted in focal acute radiation necrosis in synovial lining but no hyperplasia of synoviocytes. Later, subsynovial fibrosis became evident. White blood cell and total protein levels in the joint fluid were elevated because of intra-articular inflammation due to the acute effects of radiation. Whole knee autoradiograms showed uneven distribution of the radionuclide along the synovium and extraarticular leakage on the third day after treatment. PMID- 12088731 TI - A new and convenient method for purification of 86Y using a Sr(II) selective resin and comparison of biodistribution of 86Y and 111In labeled Herceptin. AB - A simple and rapid procedure was developed for purification of cyclotron produced 86Y via the 86Sr(p,n) 86Y reaction. A commercially available Sr(II) selective resin was used to separate 86Y from the cyclotron irradiated Sr(II) target with a recovery of the enriched Sr(II) target while yielding a 75-80% recovery of 86Y suitable for radiolabeling either proteins or peptides. To demonstrate the utility of this methodology, the anti-HER2 monoclonal antibody Herceptin was radiolabeled with the purified 86Y and compared to 111In labeled Herceptin. The biodistribution study demonstrated that 111In-Herceptin, while a suitable surrogate for 90Y in the major organs, did not parallel the uptake of 86Y Herceptin in the bone, and thus may not accurately predict the level of 90Y accumulation in the bone for clinical RIT applications. This result exemplifies the requirement of employing appropriate matched pair isotopes for imaging and therapy to insure that dosimetry considerations may be addressed accurately. PMID- 12088732 TI - Effect of vehicle on brain uptake of [11C]toluene. AB - With the goal of investigating the pharmacokinetics of the abused solvent, toluene we have adapted the rapid coupling of methyl iodide with tributylphenylstannane mediated by palladium(0) complex to the synthesis of no carrier-added [11C]toluene starting with 11CH(3)I. Two methods for purification and formulation of the tracer were developed. The first one yielded [11C]toluene dissolved in dimethylacetamide/saline solution, for the second one we adapted supercritical fluid technology where the tracer was purified using and conventional C(18) HPLC column and pure supercritical CO(2) fluid as a mobile phase operating at 2000 psi. Formulation of the tracer in cyclodextrin resulted in a significantly higher integrated uptake and distribution volume values. Additionally, we observed higher uptake and slower clearance of 11C-toluene in white matter, consistent with higher lipid content and neurotoxicological evidence indicating restricted and diffuse white matter changes in toluene abusers. This trend was observed when either DMA or cyclodextrin was used as a vehicle. It appears then, that the choice of a vehicle affected only the degree of bioavailability, but not the regional brain pharmacokinetics. Finally, we demonstrated the effect of a decreased percent difference between DV values for the studies performed on the same day, that is, test/retest variability was lower for all brain regions in beta-cyclodextrin experiments. Present results clearly demonstrate that the choice of a vehicle has a significant effect on tracer uptake and should be considered as a potential factor contributing to the pharmacokinetic measurements. PMID- 12088733 TI - Robotic preparation of Sodium Acetate C 11 Injection for use in clinical PET. AB - Sodium Acetate C 11 Injection is a radiopharmaceutical commonly used for clinical studies with positron emission tomography (PET). We have designed a fully automated robotic system for the compounding of this 20-minute half-lived tracer in the clinical setting. The system is comprised of five modular workstations that are configured in a flexible manner to accommodate all of the steps in the production of the labeled drug. The Trapping Station isolates cyclotron-produced [11C]CO(2) gas from the target and directs carbonation of methylmagnesium Grignard in diethyl ether. The Heating Station hydrolyzes the intermediate, and removes ether and unreacted [11C]CO(2) from the mixture. The Extraction Station removes ionic and organic contaminants from the drug using solid-phase extraction (AG 11A8 and C18 resin). The Filtration Station sterilizes the radiopharmaceutical, and tests membrane patency post filtration. The Assay Station measures the weight and radioactivity content of the Final Product Container, facilitating calculation of activity concentration in a remote manner. Rapid quality control methodology has also been developed that is suitable for pre-release analysis of the drug product. For a 7.5 min irradiation with a 40 microA proton beam, 223-300 mCi of Acetate C 11 Injection with purity meeting USP standards is produced within 23 min. This robotic system is a reliable method for producing Sodium Acetate C 11 Injection, USP in the clinical setting with minimal personnel exposure. Moreover, its design flexibility permits synthesis of additional (11)C- or (18)F-labeled PET tracers within the same shielded hot cell. PMID- 12088734 TI - Re: Selective in vitro labeling of white blood cells using 99mTc-labeled liposomes. PMID- 12088735 TI - Apoptosis in the rat spinal cord during postnatal development; the effect of perinatal asphyxia on programmed cell death. AB - The aim of our study was to investigate the effect of perinatal asphyxia on developmental apoptosis in the cervical and lumbar spinal cord in the neonatal rat. Perinatal asphyxia was induced by keeping pups at term in utero in a water bath at 37 degrees C for 20 min, followed by resuscitation. Effects of this treatment on developmental apoptosis were studied on postnatal days 2, 5 and 8 using terminal deoxynucleotidyl transferase (TdT)-dUTP-biotin nick end labelling (TUNEL) and caspase-3 staining. TUNEL positive cells were identified using double immunostaining. On postnatal day 2 an increase of 215% in TUNEL positive cells was detected (P=0.005) in laminae IV-VII of the lumbar spinal cord of rats which underwent perinatal asphyxia compared to controls. An increase of 55% compared to controls (P=0.03) was seen in laminae I-III of the lumbar spinal cord at postnatal day 8. TUNEL positive cells could be partly identified as microglia cells (ED1 positive) and oligodendrocytes (O4 positive). The effect of perinatal asphyxia on programmed cell death in the neonatal rat spinal cord was mainly observed in the intermediate zone and dorsal horn of the lumbar spinal cord. We conclude that perinatal asphyxia has a pronounced effect on the survival of cells in a specific region of the spinal cord and thus may have a profound effect on the development of motor networks. PMID- 12088736 TI - Intrinsic determinants of synaptic phenotype: an experimental study of abducens internuclear neurons connecting with anomalous targets. AB - The present experiments investigate the role of postsynaptic neurons in the morphological differentiation of presynaptic terminals that are formed de novo in the adult CNS. Abducens internuclear neurons in the adult cat were chosen as the experimental model. These neurons project onto the contralateral medial rectus motoneurons of the oculomotor nucleus. Abducens internuclear axon terminals were identified by their anterograde labeling with biocytin and analyzed at the electron microscopic level. To promote the formation of new synapses, two different experimental approaches were used. First, after the selective ablation of medial rectus motoneurons with ricin, abducens internuclear neurons reinnervated the neighboring oculomotor internuclear neurons. Second, after axotomy followed by embryonic cerebellar grafting, abducens internuclear axons invaded the implanted tissue and established synaptic connections in both the molecular and granule cell layer. Boutons contacting the oculomotor internuclear neurons developed ultrastructural characteristics that resembled the control synapses on medial rectus motoneurons. In the grafted cerebellar tissue, abducens internuclear axons and terminals did not resemble climbing or mossy fibers but showed similarities with control boutons. However, labeled boutons analyzed in the granule cell layer established a higher number of synaptic contacts than controls. This could reflect a trend towards the mossy fiber phenotype, although labeled boutons significantly differed in every measured parameter with the mossy fiber rosettes found in the graft. We conclude that at least for the abducens internuclear neurons, the ultrastructural differentiation of axon terminals reinnervating novel targets in the adult brain seems to be mainly under intrinsic control, with little influence by postsynaptic cells. PMID- 12088737 TI - Transient expression of juvenile-type neurocan by reactive astrocytes in adult rat brains injured by kainate-induced seizures as well as surgical incision. AB - Neurocan is one of the major chondroitin sulfate proteoglycans expressed in nervous tissues. The expression of neurocan is developmentally regulated, and full-length neurocan is detected in juvenile brains but not in adult brains. In the present study, we demonstrated by western blot analysis that full-length neurocan transiently appeared in adult rat hippocampus when it was lesioned by kainate-induced seizures. Immunohistochemical studies showed that neurocan was detected mainly around the CA1 region although the seizure resulted in neuronal cell degeneration in both the CA1 and CA3 regions of the hippocampus. Double labeling for neurocan mRNA and glial fibrillary acidic protein demonstrated that many reactive astrocytes expressed neurocan mRNA. The re-expression of full length neurocan was also observed in the surgically injured adult rat brain. In contrast, the expression of other nervous tissue chondroitin sulfate proteoglycans, such as phosphacan and neuroglycan C, was not intensified but rather was either reduced in the kainate-lesioned hippocampus or in the surgically injured cerebral cortex. These observations indicate that induction of neurocan expression by reactive astrocytes is a common phenomenon in the repair process of adult brain injury, and therefore, it can be postulated that juvenile type neurocan plays some roles in brain repair. PMID- 12088738 TI - Frequency dependent changes in mechanosensitivity of rat knee joint afferents after antidromic saphenous nerve stimulation. AB - The aim of the present study was to examine the effect of electrical saphenous nerve stimulation (14 V, 1-10 Hz) on the mechanosensitivity of rat knee joint afferents. The responses to passive joint rotations at defined torque were recorded from slowly conducting knee joint afferent nerve fibres (0.6-20.0 m/s). After repeated nerve stimulation with 1 Hz, the mechanosensitivity of about 79% of the units was significantly affected. The effects were most prominent at a torque close to the mechanical threshold. In about 46% of the examined nerve fibres a significant increase was obtained, whereas about 33% reduced their mechanosensitivity. The sensitisation was prevented by an application of 5 microM phentolamine, an alpha-adrenergic receptor blocker, together with a neuropeptide Y receptor blocker. An inhibition of N-type Ca(2+) channels by an application of 1 microM omega-conotoxin GVIA caused comparable changes of the mechanosensitivity during the electrical stimulation. Electrical nerve stimulation with higher frequencies resulted in a further reduction of the mean response to joint rotations. After stimulation with 10 Hz, there was a nearly complete loss of mechanosensitivity.In conclusion, antidromic electrical nerve stimulation leads to a frequency dependent transient decrease of the mechanosensitivity. A sensitisation was only obtained at 1 Hz, but this effect may be based on the influence of sympathetic nerve fibres. PMID- 12088739 TI - Feeding-induced decrease in extracellular glutamate level in the rat nucleus accumbens: dependence on glutamate uptake. AB - In vivo microdialysis combined with high-performance liquid chromatography and electrochemical detection was used to monitor extracellular glutamate levels in the medial nucleus accumbens of Sprague-Dawley rats during their feeding behaviour. Consumption of a palatable new diet or a diet to which rats were previously exposed caused a decrease in extracellular level of glutamate in the nucleus accumbens during and after feeding. The presentation of an inedible object (a piece of rubber) instead of the expected food caused a marked increase in extracellular glutamate levels. In contrast, if the piece of rubber was presented to rats that did not expect food delivery, the extracellular level of glutamate remained unchanged during the rubber presentation. The feeding-induced decrease in the extracellular glutamate level did not depend on food deprivation and was completely prevented by intraaccumbal infusions through the dialysis probe of 10 mM D,L-threo-beta-hydroxyaspartate (a glutamate uptake inhibitor). Intraaccumbal infusions of 10 microM S-(-)-raclopride L-tartrate (a D2/D3 dopamine receptor antagonist) or 1 microM tetrodotoxin (a voltage-dependent Na(+) channel blocker) also completely reversed the decrease in extracellular glutamate level in response to food intake. The D1/D5 dopamine receptor antagonist SCH 23390 (10 microM) administered into the nucleus accumbens had no significant effect on the feeding-induced decrease in extracellular glutamate level. From the data obtained we suggest that the decrease in the extracellular level of glutamate in the medial nucleus accumbens in response to feeding appears to arise from a temporal increase in glutamate uptake that is probably operated by dopamine inputs to the nucleus accumbens via D2/D3 receptors. Our findings also suggest that the dissociation between the expected biological value of a presented object and the reality might be an important determinant for regulation of glutamate release in this brain area during feeding behaviour. PMID- 12088740 TI - A high-fat, refined sugar diet reduces hippocampal brain-derived neurotrophic factor, neuronal plasticity, and learning. AB - We have investigated a potential mechanism by which a diet, similar in composition to the typical diet of most industrialized western societies rich in saturated fat and refined sugar (HFS), can influence brain structure and function via regulation of neurotrophins. We show that animals that learn a spatial memory task faster have more brain-derived neurotrophic factor (BDNF) mRNA and protein in the hippocampus. Two months on the HFS diet were sufficient to reduce hippocampal level of BDNF and spatial learning performance. Consequent to the action of BDNF on synaptic function, downstream effectors for the action of BDNF on synaptic plasticity were reduced proportionally to BDNF levels, in the hippocampus of rats maintained on the HFS diet between 2 and 24 months. In particular, animals maintained on the HFS diet showed a decrease in levels of: (i) synapsin I mRNA and protein (total and phosphorylated), important for neurotransmitter release; (ii) cyclic AMP-response element-binding protein (CREB) mRNA and protein (total and phosphorylated); CREB is required for various forms of memory and is under regulatory control of BDNF; (iii) growth-associated protein 43 mRNA, important for neurite outgrowth, neurotransmitter release, and learning and memory. Diet-related changes were specific for the hippocampus consequent to its role in memory formation, and did not involve neurotrophin-3, another member of the neurotrophin family. Our results indicate that a popularly consumed diet can influence crucial aspects of neuronal and behavioral plasticity associated with the function of BDNF. PMID- 12088741 TI - Synaptically driven spikes and long-term potentiation in neocortical layer 2/3. AB - Recently, variation upon a well-established hippocampal model has given rise to a new paradigm in which the strength of synaptic inputs to neocortical layer 2/3 is estimated in vitro by recording synaptically driven extracellular potentials elicited there by electrical stimulation applied to underlying layer 4. The analysis of these potentials is commonly based upon an assumption that postsynaptic spiking has played no significant role in their generation. Here, we have tested this assumption by quantifying in rats (using data obtained by cell attached recording) the rate at which unit spikes are elicited in layer 2/3 under commonly used conditions of stimulation and recording. We found that spike responses were regularly elicited at the same latencies as field potential peaks and the rising phases of intracellularly recorded synaptic currents, and the incidence of such spiking (the fractional rate of cells spiking versus cells sampled) was sufficient to give this higher-order activity a major role in determining response amplitudes. We then analyzed layer 2/3 waveform characteristics before and after inducing long-term potentiation (LTP) by theta burst stimulation (TBS) and found that the induction of LTP succeeded only when the initial response included a strong spike component. We further observed that LTP expression was always accompanied by a pronounced enhancement of such components. Our data suggest that, unlike in hippocampal CA1, LTP elicited by TBS in this neocortical paradigm depends upon modification of synaptically driven spike activity, through either enhanced synchronization of unitary responses, the recruitment of additional responding units, or both. This potentiation of the spike response could arise (as previously proposed) through an increase in the efficacy of synapses mediating projection from layer 4 to 2/3, but other mechanisms may also contribute, such as modification of short-range recurrent connections within layer 2/3, which are likely to play an important role in defining local-network cell ensembles. PMID- 12088742 TI - Uptake and pathogenic effects of amyloid beta peptide 1-42 are enhanced by integrin antagonists and blocked by NMDA receptor antagonists. AB - Many synapses contain two types of receptors - integrins and N-methyl-D-aspartate (NMDA) receptors - that have been implicated in peptide internalization. The present studies tested if either class is involved in the uptake of the 42 residue form of amyloid beta peptide (Abeta1-42), an event hypothesized to be of importance in the development of Alzheimer's disease. Cultured hippocampal slices were exposed to Abeta1-42 for 6 days in the presence or absence of soluble Gly Arg-Gly-Asp-Ser-Pro, a peptide antagonist of Arg-Gly-Asp (RGD)-binding integrins, or the disintegrin echistatin. Abeta uptake, as assessed with immunocytochemistry, occurred in 42% of the slices incubated with Abeta peptide alone but in more than 80% of the slices co-treated with integrin antagonists. Uptake was also found in a broader range of hippocampal subfields in RGD-treated slices. Increased sequestration was accompanied by two characteristics of early stage Alzheimer's disease: elevated concentrations of cathepsin D immunoreactivity and activation of microglia. The selective NMDA receptor antagonist D-(-)-2-amino-5-phosphonovalerate completely blocked internalization of Abeta, up-regulation of cathepsin D, and activation of microglia. Our results identify two classes of receptors that cooperatively regulate the internalization of Abeta1-42 and support the hypothesis that characteristic pathologies of Alzheimer's disease occur once critical intraneuronal Abeta concentrations are reached. PMID- 12088743 TI - Osmotic stress increases brain-derived neurotrophic factor messenger RNA expression in the hypothalamic supraoptic nucleus with differential regulation of its transcripts. Relation to arginine-vasopressin content. AB - We have shown that osmotic stress increases brain-derived neurotrophic factor (BDNF) mRNA in the supraoptic nucleus and that this increase seems to be determined by the high expression of transcripts containing exon I. The paraventricular nucleus is another hypothalamic neuronal subset where BDNF mRNA is also sensitive to osmotic stress stimulation. In this nucleus, transcripts containing exon I were not modified but only those containing exon II. By contrast, transcripts containing exon III did not exhibit any variation in our experimental conditions. The presence of BDNF mRNA in both paraventricular and supraoptic hypothalamic nuclei was recently reported. These nuclei are extremely sensitive to osmotic stimuli and their neurons secrete oxytocin and arginine vasopressin in the posterior pituitary gland. This study was thus designed to investigate the possible involvement of BDNF in the response of supraoptic nucleus to osmotic stress stimulus. Osmotic stress was induced by hypertonic saline injection (1.35% NaCl) administered to animals 3 h before analysis. We used non-isotopic in situ hybridization to study the expression of BDNF mRNA and its transcripts with antisense riboprobes on histological brain sections, including paraventricular and supraoptic nuclei from control and osmotic stress stimulated animals. To investigate a possible correlation between the expression of BDNF mRNA and arginine-vasopressin, the peptide content was analyzed by immunohistochemistry in both paraventricular and supraoptic nuclei at two different times after hyperosmotic injection. The results showed that BDNF mRNA expression preceded the arginine-vasopressin increase. In addition, on serial adjacent histological sections of supraoptic nucleus (10 microm), both BDNF and arginine-vasopressin mRNAs were visualized by isotopic in situ hybridization and the images were overlaid, showing that almost all of the hybridization signals were overlapped. Taken together our results are in keeping with the hypothesis that activation of the different BDNF promoters seems to be region-specific. Besides, the temporal correlation between both BDNF mRNA expression and arginine vasopressin content, as well as the morphological vicinity between their respective producing cells in the supraoptic nucleus, suggest an autocrine or paracrine action for this neurotrophin in the regulation of arginine-vasopressin secretion. PMID- 12088744 TI - Blockade of nigral and pallidal opioid receptors suppresses vacuous chewing movements in a rodent model of tardive dyskinesia. AB - Chronic neuroleptic treatment leads to the development of tardive dyskinesia in 20-30% of patients. While the pathogenesis of tardive dyskinesia remains elusive, altered opioid peptide function in striatal projection pathways of the basal ganglia has been implicated. Using a rodent model of vacuous chewing movements induced by chronic neuroleptic administration, we investigated regional involvement of opioid transmission in tardive dyskinesia. We examined the role of dynorphin in the direct striatonigral pathway by infusing nor-binaltorphimine, a selective kappa opioid receptor antagonist, into the substantia nigra pars reticulata. As well, infusions of naloxone (a non-specific opioid receptor antagonist), D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr amide (CTOP; a mu opioid receptor antagonist) or naltrindole (a delta opioid receptor antagonist) into the globus pallidus were used to establish the contribution of the striatopallidal pathway. Chronic fluphenazine treatment (25 mg/kg i.m. every 3 weeks for 18 weeks) resulted in a robust increase in vacuous chewing movements. Infusion of nor-binaltorphimine (5.0 nmol) into the substantia nigra pars reticulata significantly attenuated vacuous chewing movements. Infusion of naloxone (0.5 and 2.0 nmol) into the globus pallidus also significantly attenuated vacuous chewing. Infusion of naltrindole into the globus pallidus blocked vacuous chewing at all doses administered (0.5, 1.0, 2.0 nmol) while CTOP was only effective at the two higher doses. From these results we suggest that increases in dynorphin in the direct striatonigral pathway and enkephalin in the indirect striatopallidal pathway following chronic neuroleptic administration are both likely to contribute to tardive dyskinesia. PMID- 12088745 TI - Glutamate-induced currents reveal three functionally distinct NMDA receptor populations in rat dorsal horn - effects of peripheral nerve lesion and inflammation. AB - The importance of the N-methyl-D-aspartate (NMDA) receptor in various painful conditions is well established. The effects of peripheral nerve lesion or joint inflammation, as models of different pain states, on NMDA receptor-mediated currents and NMDA receptor subunit mRNA expression were therefore studied in acutely dissociated neurones from the rat spinal cord dorsal horn. In the neuronal population from control rats, all four NR2 subunits and both NR1 splice variants assayed were detected. A majority of neurones expressed mRNA for more than one NR2 subunit, and some neurones expressed all four NR2 subunits as well as both NR1 splice variants. The NR2B subunit was the most commonly expressed, while the NR2C was the rarest. Following nerve lesion, fewer neurones expressed NR2A compared to the control. The dose-response curve for glutamate-evoked NMDA receptor-mediated currents in the neurones was best described by a three component fit, suggesting that three functionally distinct NMDA receptor populations are present in the dorsal horn. Minor changes in the dose-response curve after nerve lesion could not be ascribed with certainty to the lesion. Changes in other parameters of NMDA receptor-mediated currents were observed neither after nerve lesion nor after joint inflammation. In summary, the present work demonstrates that single dorsal horn neurones express mRNA for several NMDA receptor subunits. The glutamate dose-response curves indicate that there are three major types of NMDA receptors present in dorsal horn neurones. We also report a reduced expression of NR2A following peripheral nerve lesion. PMID- 12088746 TI - Nitration is a mechanism of regulation of the NMDA receptor function during hypoxia. AB - The present study tested the hypothesis that nitration is a mechanism of hypoxia induced modification of the N-methyl-D-aspartate (NMDA) receptor. To test this hypothesis the effect of hypoxia on the nitration of the NR1, NR2A and NR2B subunits of the NMDA receptor was determined. Furthermore, the effect of administration of a nitric oxide synthase (NOS) inhibitor, N-nitro-L-arginine (NNLA) on the hypoxia-induced nitration of the NMDA receptor subunits as well as the NMDA receptor-mediated Ca2+ influx, an index of NMDA receptor-ion channel function, were determined in cortical synaptosomes. Studies were performed in newborn piglets divided into normoxic, hypoxic and hypoxic-NNLA groups. Hypoxia was induced by decreasing the FiO(2) to 0.07-0.09 for 60 min. Cerebral tissue hypoxia was confirmed by determining the levels of high energy phosphates ATP and phosphocreatine. Nitration of the NMDA receptor subunits was determined by immunoprecipitation using specific antibodies and western blot analysis. NMDA receptor-ion channel-mediated Ca2+ influx was determined using 45Ca2+. There was a significant increase in the nitrated NR1, NR2A and NR2B subunits following hypoxia: 104+/-11 vs. 275+/-18 optical density (OD)xmm(2) for NR1 (P<0.05), 212+/ 36 vs. 421+/-16 ODxmm(2) for NR2A (P<0.05) and 246+/-44 vs. 360+/-26 ODxmm(2) for NR2B (P<0.05). This increase in nitrated NR1, NR2A and NR2B subunits of the NMDA receptor was prevented by the administration of NNLA prior to hypoxia (NR1 160+/ 19, P=NS, NNLA vs. normoxic; NR2A 304+/-49, P=NS, NNLA vs. normoxic, and NR2B 274+/-19, P=NS, NNLA vs. normoxic). The increase in nitration of the NR1, NR2A and NR2B subunits of the NMDA receptor increased as a function of decreased cerebral high-energy phosphates, ATP and phosphocreatine, during hypoxia. Furthermore, NOS blockade prior to hypoxia resulted in prevention of the hypoxia induced increase in NMDA receptor-mediated Ca2+ influx. Our results demonstrate that hypoxia results in increased nitration of the NMDA receptor subunits and that administration of an NOS inhibitor prior to hypoxia prevents the hypoxia induced nitration of the NMDA receptor subunits as well as the hypoxia-induced increase in NMDA receptor-mediated Ca2+ influx. We conclude that nitration is a mechanism of modification of the NMDA receptor function during hypoxia in the newborn piglet brain. PMID- 12088747 TI - Early chronic aluminium exposure impairs long-term potentiation and depression to the rat dentate gyrus in vivo. AB - As an important neurotoxin, aluminium can cause cognitive dysfunctions and mental diseases. Previous studies have reported that aluminium impaired long-term potentiation (LTP) in vivo and in vitro. Here, we utilise two models of synaptic plasticity, LTP and long-term depression (LTD) to study the effects of aluminium on synaptic plasticity in vivo. Neonatal Wistar rats were chronically exposed to aluminium from birth to weaning via the milk of dams fed with 0.3% aluminium chloride solution. Excitatory postsynaptic potential (EPSP) and population spikes (PS) were recorded from the dentate gyrus (DG) of adult rats by electrically stimulating the perforant path. THE FOLLOWING RESULTS WERE OBTAINED: (1) The input/output function indicated that, as compared to controls, aluminium increased the baseline amplitude of the PS, but decreased the baseline slope of EPSP. (2) Aluminium significantly prevented LTD in PS (controls: 77.36+/-6.7%, n=7; aluminium-exposed: 102.01+/-9.1%, n=7; P<0.05) and decreased the LTD amplitude in EPSP (controls: 76.61+/-4.1%, n=7; aluminium-exposed: 94.31+/-7.9% n=7, P<0.05). (3) Aluminium reduced the amplitude of LTP in both PS (controls: 190+/-16.1%, n=7; aluminium-exposed: 135+/-9.7%, n=7; P<0.05) and EPSP (control: 132+/-9.3%, n=7; aluminium-exposed: 115+/-10.6%, n=7; P<0.05). As for LTD and LTP, PS was impaired more seriously than EPSP in aluminium-exposed rats. (4) Aluminium exposure decreased the paired-pulse facilitation (PPF) of PS at 30-150 ms interpulse interval (IPI), and reduced 93.5% of PPF at 80 ms IPI in PS (controls: 243.4+/-39.8%, n=7; aluminium-exposed: 149.9+/-12.3%, n=7). There was no significant difference in EPSP of PPF. From these results we conclude that aluminium exposure in neonatal rats thus reduces the amplitude of LTP and PPF and blocks the induction of LTD in the DG. We suggest that aluminium affects both presynaptic and postsynaptic mechanisms of synaptic transmission. PMID- 12088748 TI - Expression of macrophage colony-stimulating factor and its receptor in microglia activation is linked to teratogen-induced neuronal damage. AB - Prenatal exposure to teratogen agents is linked to the pathogenesis of neurodevelopment disorders, but the mechanisms leading to the neurodevelopmental disturbance are poorly understood. To elucidate this, an in vitro model of microglial activation induced by neuronal injury has been characterized. In this connection, exposure of primary microglial cells to the conditioned medium from the neuronal damage induced by teratogen, cyclophosphamide, is accompanied by a reactive microgliosis as assessed by reverse transcription-polymerase chain reaction, enzyme-linked immunosorbent assay, lectin histochemistry, double labeling immunohistochemistry and in situ hybridization. Our results showed that reactive microglia were capable of releasing various cytokines such as tumor necrosis factor-alpha, interleukin-1, interleukin-6, transforming growth factor beta and nitric oxide. Also, we have shown that macrophage colony-stimulating factor (M-CSF) was in fact produced by the reactive microglia. Concomitant to this was the increased expression of M-CSF receptor in these cells following the teratogen-induced neuronal injury. The up-regulation of M-CSF receptor suggests that the cells are capable of responding to self-derived M-CSF in an autocrine fashion. Results with antibody neutralization further suggest that microglial proinflammatory response, as manifested by cytokine expression in culture, is mediated by M-CSF, which acts as a molecular signal that initiates a microglial reaction. We therefore suggest that microglial activation following cyclophosphamide treatment is not only a response to the neuronal damage, but is also a cause of the damage during pathogenesis of neurodevelopment disorders. To this end, the increased expression of M-CSF and its receptor on microglia would be directly linked to the active cell proliferation and proinflammatory response in the teratogen-induced injury. PMID- 12088749 TI - Non-specific olfactory aversion induced by intrabulbar infusion of the GABA(A) receptor antagonist bicuculline in young rats. AB - On postnatal day 12, young rats show an aversion to an odor to which they had been exposed along with presentations of foot shock on postnatal day 11. The acquisition of this aversive learning involves and requires disinhibition of the mitral/tufted cells induced by centrifugal noradrenergic activation during somatosensory stimulation. This olfactory learning is established only for the odor to which the rat has been exposed during conditioning. Infusion of the GABA(A) receptor antagonist bicuculline at a high dose (2.0 nmol/each olfactory bulb) into the olfactory bulb in the presence of an odor is capable of developing olfactory aversive responses without somatosensory stimulation in young rats. The purpose of this study is to characterize the properties of bicuculline-induced aversive responses. In contrast to the odor specificity of aversive learning produced by odor-shock conditioning, bicuculline-induced aversive responses lack odor specificity. Namely, bicuculline infusion in the presence of a citral odor results, in a dose-dependent manner, in subsequent aversive responses to strange odors (benzaldehyde and vanillin) that have never been presented. Moreover, bicuculline infusion alone is sufficient to produce dose-dependent aversive responses to strange odors (citral, benzaldehyde and geraniol). From these results we suggest that disinhibition of mitral/tufted cells from granule cells by bicuculline infusion makes young rats aversive to strange odors non specifically, as if the rats had learned the odor aversion as a result of odor exposure paired with foot shock. Different mechanisms of disinhibition of the mitral/tufted cells may underlie both the pharmacological manipulation and noradrenergic activation by somatosensory stimulation. PMID- 12088750 TI - Changes in basal and cocaine-evoked extracellular dopamine uptake and release in the rat nucleus accumbens during early abstinence from cocaine: quantitative determination under transient conditions. AB - Despite an abundance of studies on mechanisms of behavioral sensitization, considerable uncertainty exists as to whether alterations in dopamine neurotransmission underlie the exacerbated behavioral effects of cocaine observed during the early stages of abstinence. One of the factors contributing to the uncertainty and controversy may be the limitations in utilized measurement techniques (mostly conventional microdialysis). The techniques of quantitative microdialysis under transient conditions and rotating disk electrode voltammetry were used to characterize basal dopamine dynamics as well as time-related changes in extracellular dopamine concentrations and dopamine uptake that occur in response to an acute drug challenge in control animals and animals with previous history of cocaine. Basal extracellular dopamine concentrations were unaltered on abstinence day 3 from repeated cocaine administration (5 days, 20 mg/kg, i.p.). The extraction fraction of dopamine, an indirect measure of dopamine uptake, was significantly lower in cocaine-sensitized animals relative to controls. These two facts, taken together, suggest that basal dopamine release is depressed in cocaine-sensitized animals on abstinence day 3. At the same time, a cocaine challenge decreased the extraction fraction and increased the extracellular dopamine concentration in both experimental groups. The magnitude of the increase in extracellular dopamine concentration was greater in cocaine-sensitized animals, while the ability of cocaine to decrease the extraction fraction was unaltered, suggesting that the increase in extracellular dopamine concentration reflects an increase in drug-evoked dopamine release. Moreover, cocaine pretreated rats demonstrated greater depolarization-induced dopamine release and the ability of dopamine D(2) receptor agonist, quinpirole, to inhibit release was decreased in these animals. These data demonstrate that a cocaine treatment regimen resulting in behavioral sensitization is associated with a reduction in basal dopamine release, an enhancement in both cocaine and K(+)-evoked dopamine release, and a subsensitivity of dopamine D(2) autoreceptors that regulate dopamine release in the nucleus accumbens. PMID- 12088751 TI - Developmental regulation and neuronal expression of the cellular disintegrin ADAM11 gene in mouse nervous system. AB - ADAM11 is the prototype member of the predominantly CNS-associated clade of the ADAM metalloprotease-disintegrins that has been implicated in neural adhesion and axon guidance. The present study describes the spatiotemporal expression pattern of the ADAM11 gene in adult and developing mouse, and identifies the cells expressing the gene. In the adult CNS, ADAM11 mRNA was present throughout the forebrain, including different cortical fields and diencephalic nuclei. In brainstem, low to moderate expression was detected in certain midbrain nuclei, while several pontine and medullary nuclei showed a very strong signal. High expression was observed in the cerebellar cortex and spinal cord. In addition, ADAM11 was expressed in ganglia of the peripheral nervous system (PNS), retinae, testes, liver, and at lower levels in epidermal and mucosal epithelia, kidney, and salivary gland. The expression was localized to neurons in all examined CNS and PNS subfields. During pre- and perinatal development, ADAM11 was differentially expressed both in the developing PNS and CNS, as well as in heart, kidney, eyes, and brown fat. The present results suggest a widespread involvement of ADAM11 in neuron-neuron or neuron-glial cell interactions during development as well as in the adult nervous system. They provide novel complementary information to recently accumulated data on CNS integrin gene expression and offer useful clues for further studies of the neural functions of ADAMs and integrins. PMID- 12088752 TI - Serotonin(3) receptor stimulation in the nucleus tractus solitarii activates non catecholaminergic neurons in the rat ventrolateral medulla. AB - The present study was performed to determine whether or not the increased arterial pressure triggered by 5-HT(3) receptor stimulation in the nucleus tractus solitarii and underlain by a sympathoexcitation is associated with the activation of ventromedullary cells known to be involved in vascular regulation, i.e. the C1 and A1 catecholaminergic cells. For this purpose, double immunohistochemical labeling for tyrosine hydroxylase and c-fos protein was performed all along the ventrolateral medulla after microinjection of 1-(m chlorophenyl)-biguanide, a selective and potent 5-HT(3) receptor agonist, into the nucleus tractus solitarii of alpha-chloralose/urethane-anaesthetized rats. This treatment produced a significant elevation of arterial pressure ( approximately +35 mm Hg). Concomitantly, a significant increase in the number of c-fos expressing neurons was observed in the rostral ventrolateral medulla (+63%), in particular in its most anterior part (+78%), and in the medullary region surrounding the caudal part of the facial nucleus (+91%). Retrograde labeling with gold-horseradish peroxidase complex showed that at least some of these activated c-fos expressing cells project to the spinal cord. However, the number of double-stained neurons, i.e. c-fos and tyrosine hydroxylase positive neurons, did not increase at any level of the ventrolateral medulla. In contrast, under the same alpha-chloralose/urethane anesthesia, systemic infusion of sodium nitroprusside appeared to produce a hypotension and a marked increase in the density of such double c-fos and tyrosine hydroxylase expressing cells in the rostral ventrolateral medulla and the caudal medullary region surrounding the caudal part of the facial nucleus. These data indicate that medullary catecholaminergic C1 and A1 neurons are not involved in the pressor effect elicited by 5-HT(3) receptor stimulation in the nucleus tractus solitarii. However, this 5-HT(3) receptor-mediated effect is clearly associated with the excitation of (non-catecholaminergic) neurons within the pressor region of the ventral medulla. PMID- 12088753 TI - Forebrain-specific promoter/enhancer D6 derived from the mouse Dach1 gene controls expression in neural stem cells. AB - Drosophila dachshund is involved in development of eye and limbs and in the development of mushroom bodies, a brain structure required for learning and memory in flies. Its mouse homologue mDach1 is expressed in various embryonic tissues, including limbs, the eye, the dorsal spinal cord and the forebrain. We have isolated a forebrain-specific 2.5-kb enhancer element termed D6 from the mouse mDach1 gene and created D6-LacZ and D6-green fluorescent protein (GFP) reporter gene mouse lines. In embryonic stages, the D6 enhancer activity is first detected at embryonic day 10.5 in scattered cells of the outbuldging cortical vesicles. By embryonic day 12.5, D6 activity expands throughout the developing neocortex and the hippocampus. In the adult mouse brain, D6 enhancer is active in neurons of the cortical plate, in the CA1 layer of the hippocampus and in cells of the subventricular zone and the ventricular ependymal zone. Adult mice also show D6 activity in the olfactory bulb and in the mamillary nucleus. Cultured D6 positive cells, which were derived from embryonic and postnatal brains, show characteristics of neural stem cells. They form primary and secondary neurospheres that differentiate into neurons and astrocytes as examined by cell specific markers.Our results show that D6 enhancer exerts highly tissue-specific activity in the neurons of the neocortex and hippocampus and in neural stem cells. Moreover, the fluorescence cell sorting of D6-GFP cells from embryonic and postnatal stages allows specific selection of primary neural progenitors and their analysis. PMID- 12088754 TI - Injection of brain-derived neurotrophic factor in the rostral ventrolateral medulla increases arterial blood pressure in anaesthetized rats. AB - Brain-derived neurotrophic factor (BDNF) is a unique neurotrophin which not only supports the development of neurons but also modulates the synaptic activity in a number of neuronal systems. BDNF is synthesized in neurons, anterogradely transported and released from nerve terminals and exerts acute effects on synaptic transmission in both peripheral and central nervous systems. Previous studies have shown that BDNF is distributed in several groups of neurons in the brain stem which regulate cardiovascular functions. Here we showed that injection of BDNF (40-400 ng/100 nl) into the rostral ventrolateral medulla resulted in a significant increase in arterial blood pressure (Delta35.5+/-3.5 mmHg) in rats. The duration of change in blood pressure was 145+/-40 s with a latency of 3-5 s. There was no significant effect on the heart rate. The injection of glutamate as a positive control also triggered an increase in blood pressure. Injection of phosphate-buffered saline as a control or the same amount of nerve growth factor did not cause significant changes in blood pressure in different preparations. Immunohistochemistry showed that the nerve terminals immunoreactive for BDNF were localized in several brain stem regions and terminate around spinal projection neurons in the rostral ventrolateral medulla. Neurons in the rostral ventrolateral medulla can uptake exogenous BDNF and express the high affinity receptor trkB. From these results we suggest that BNDF in the medulla may play a role in the regulation of blood pressure. PMID- 12088755 TI - Chronic hypoxia up-regulates fibroblast growth factor ligands in the perinatal brain and induces fibroblast growth factor-responsive radial glial cells in the sub-ependymal zone. AB - A number of signaling molecules have been implicated in the acute response to hypoxia/ischemia in the adult brain. In contrast, the reaction to chronic hypoxemia is largely unexplored. We used a protocol of chronic hypoxia in rat pups during the first three postnatal weeks, encompassing the period of cellular plasticity in the cerebral cortex. We find that the levels of fibroblast growth factor 1 (FGF1) and FGF2, two members of the FGF family, increase after 2 weeks of chronic hypoxia. In contrast, members of the neurotrophin family are unaffected. FGF2 is normally expressed in the nucleus of mature, glial fibrillary acidic protein (GFAP)-containing astrocytes. Under hypoxia, most FGF2-containing cells do not express detectable levels of GFAP, suggesting that chronic low O(2) induces their transformation into more immature glial phenotypes. Remarkably, hypoxia promotes the appearance of radial glia throughout the sub-ventricular and ependymal zones. Most of these cells express vimentin and brain lipid binding protein. A subset of these radial glial cells expresses FGF receptor 1, and are in close contact with FGF2-positive cells in the sub-ventricular zone. Thus, FGF receptor signaling in radial glia may foster cell genesis after chronic hypoxic damage. From the results of this study we suggest that after the chronic exposure to low levels of oxygen during development, the expression of radial glia increases in the forebrain periventricular region. We envision that astroglia, which are the direct descendants of radial glia, are reverting back to immature glial cells. Alternatively, hypoxia hinders the normal maturation of radial glia into GFAP-expressing astrocytes. Interestingly, hypoxia increases the levels of expression of FGF2, a factor that is essential for neuronal development. Furthermore, chronic hypoxia up-regulated FGF2's major receptor in the periventricular region. Because radial glia have been suggested to play a key role in neurogenesis and cell migration, our data suggests that hypoxia-induced FGF signaling in radial glia may represent part of a conserved program capable of regenerating neurons in the brain after injury. PMID- 12088756 TI - Cortical expression of endothelin receptor subtypes A and B following middle cerebral artery occlusion in rats. AB - This work aimed to define the spatial expression of endothelin A (ET(A)) and B (ET(B)) receptors in the cerebral cortex after permanent middle cerebral artery occlusion (MCAO) and to identify the phenotype of cells expressing ET(A) and ET(B) receptors. Cortical expression of ET(A) and ET(B) receptors was determined at the mRNA level by semi-quantitative reverse transcription-polymerase chain reaction and at the protein level by immunofluorescence staining, 12, 24 and 72 h after MCAO. Cells expressing endothelin receptors were phenotyped by double labelling with antibodies, anti-protein gene product (PGP9.5) and anti-ED1, towards neurons and activated microglia/macrophages, respectively. Both ET(A) and ET(B) receptor mRNA expressions increased significantly in the ipsilateral cortex in a time-dependent manner after MCAO. Robust expression of ET(A) receptors was noted in most neurons of the ischemic core and in several neurons in laminae 3 and 4 of the peri-infarct region 24 and 72 h after MCAO. ET(B) receptor immunoreactivity was observed in activated microglia/macrophages, beginning 24 h after MCAO. These results provide the first evidence that the action of endothelin during ischemia may be mediated by neuronal ET(A) receptors and activated microglia/macrophage ET(B) receptors. This differential localization of ET(A) and ET(B) receptors suggests that endothelin is involved in some complex neuron-glial interactions in addition to its vascular modulatory activity during ischemia. PMID- 12088757 TI - Local circuit plasticity in the rat dentate gyrus: characterization and aging related impairment. AB - We have used frequency-dependent inhibition, a form of short-term plasticity mediated by the activation of inhibitory interneurons, to characterize in vivo alterations in local circuit activity and plasticity in the dentate gyrus of the anesthetized rat. The application of the GABA-A receptor blocker, bicuculline, induced a transient reduction in frequency-dependent inhibition, indicating that this form of local circuit activity is GABA-mediated. Delivering theta burst stimulation to the perforant pathway of the hippocampus induced long-term potentiation of the population excitatory post-synaptic potential, reflecting the potentiation of the perforant path-dentate gyrus granule cell synapses. Concomitantly, theta burst stimulation caused a lasting reduction in frequency dependent inhibition. In aged rats, long-term potentiation could be induced to the same level as in young rats, but while in young rats frequency-dependent inhibition was concomitantly reduced, frequency-dependent inhibition in the old rats did not show this form of plasticity. Our results indicate that theta burst stimulation induces a form of local circuit plasticity independently of its known capacity to induce synaptic plasticity, and that this form of local circuit plasticity is compromised in aging. Based on these results we propose a potential role for plasticity at the level of the local circuit in learning and memory. PMID- 12088758 TI - Prolactin and schizophrenia: clinical consequences of hyperprolactinaemia. AB - Prolactin is a polypeptide hormone that is synthesized and secreted from specialised cells of the anterior pituitary gland, known as lactotrophs. The hormone was given it's name because extracts from the bovine pituitary gland caused growth of the crop sac and stimulated the elaboration of crop milk in pigeons, and promoted lactation in rabbits. Although prolactin is best known for the multiple effects it exerts on the mammary gland, it has over 300 separate biological activities not represented by its name. It sub serves multiple roles in reproduction other than lactation and is an important modulator of homeostasis in the mammalian organism. Hence Bern and Nicoll suggested renaming it "omnipotin or versatilin". Schizophrenia is a severe psychiatric disorder that affects approximately one percent of the population worldwide. It is well established that traditional typical anti-psychotics elevate prolactin levels. It is also agreed that the serum prolactin concentration is not elevated in patients with schizophrenia who are not receiving anti-psychotic medication. Hyperprolactinaemia has direct effects on the brain and on other organs. Direct consequences include galactorrhoea. Indirect consequences of hyperprolactinaemia include oligomenorrhoea and amenorrhoea, erratic or absent ovulation, sexual dysfunction, reduced bone mineral density and cardiovascular disease. With the advent of prolactin sparing anti-psychotics, ample consideration needs to be given to the physiological consequences of hyperprolactinaemia in schizophrenic patients. In this paper we will examine molecular biology, secretion and physiology of prolactin. The consequences of hyperprolactinaemia in humans including effects on fertility, sexual dysfunction, bone mineral density, cardiovascular disease, changes in psychopathology and movement disorders will be reviewed. The literature on the association between schizophrenia, anti-psychotic medication and hyperprolactinaemia and more specifically on the consequences of this hyperprolactinaemia in schizophrenic patients will also be reviewed. PMID- 12088759 TI - Effects of docosahexaenoic acid on calcium pathway in adult rat cardiomyocytes. AB - In this study we examined the effect of polyunsaturated fatty acids (PUFAs), in particular of docosahexaenoic acid (DHA), on calcium homeostasis in isolated adult rat cardiomyocytes exposed to KCl, ET-1 and anoxia. Free [Ca(2+)](i) in rat cardiomyocytes was 135.7 +/- 0.5 nM. Exposure to 50 mM KCl or 100 nM ET-1 resulted in a rise in free [Ca(2+)](i) in freshly isolated cells (465.4 +/- 15.6 nM and 311.3 +/- 12.6 nM, respectively) and in cultured cells (450.8 +/- 14.8 nM and 323.5 +/- 14.8 nM respectively). An acute treatment (20 minutes) with 10 microM DHA significantly reduced the KCl- and ET-1-induced [Ca(2+)](i) increase (300.9 +/- 18.1 nM and 232.08 +/- 11.8 nM, respectively). This reduction was greater after chronic treatment with DHA (72 h; 257.7 +/- 13.08 nM and 192.18 +/- 9.8 nM, respectively). Rat cardiomyocytes exposed to a 20 minute superfusion with anoxic solution, obtained by replacing O(2) with N(2) in gas mixture, showed a massive increase in cytosolic calcium (1200.2 +/- 50.2 nM). Longer exposure to anoxia induced hypercontraction and later death of rat cardiomyocytes. Preincubation with DHA reduced the anoxic effect on [Ca(2+)](i) (498.4 +/- 7.3 nM in acute and 200.2 +/- 12.2 nM in chronic treatment). In anoxic conditions 50 mM KCl and 100 nM ET-1 produced extreme and unmeasurable increases of [Ca(2+)](i.) Preincubation for 20 minutes with DHA reduced this phenomenon (856.1 +/- 20.3 nM and 782.3 +/- 7.6 nM, respectively). This reduction is more evident after a chronic treatment with DHA (257.7 +/- 10.6 nM and 232.2 +/- 12.5 nM, respectively). We conclude that in rat cardiomyocytes KCl, ET-1 and anoxia interfered with intracellular calcium concentrations by either modifying calcium levels or impairing calcium homeostasis. Acute, and especially chronic, DHA administration markedly reduced the damage induced by calcium overload in those cells. PMID- 12088760 TI - VIP-ellipticine derivatives inhibit the growth of breast cancer cells. AB - The effects of vasoactive intestinal peptide (VIP)-ellipticine (E) derivatives were investigated on breast cancer cells. VIP-ALALA-E and VIP-LALA-E inhibited 125I-VIP binding to MCF-7 cells with an IC(50) values of 1 and 0.2 microM respectively. VIP-ALALA-E and VIP-LALA-E caused elevation of cAMP in MCF-7 cells with ED(50) values of 1 and 0.1 microM. VIP-LALA-E caused increased c-fos mRNA in MCF-7 cells. Radiolabeled VIP-LALA-E was internalized at 37 degrees C and delivered the cytotoxic E into MCF-7 cells. VIP-LALA-E inhibited the clonal growth of MCF-7 cells, decreased cell viability based on trypan blue exclusion and reduced 35S-methionine uptake. These results indicate that VIP-E derivatives function as breast cancer VPAC(1) receptor agonists which inhibit MCF-7 cellular viability. PMID- 12088761 TI - Effect of experimental hypoalbuminemia on the plasma protein binding of tolmetin. AB - The purpose of this work was to study tolmetin plasma protein binding in an experimental model of hypoalbuminemia in the rat. Hypoalbuminemia was produced by repetitive plasmapheresis, achieving a 26.2 +/- 4.6% reduction in albumin circulating levels. Rats then received a 100 mg/kg oral tolmetin dose. Control rats received oral tolmetin 10, 56 or 100 mg/kg. Tolmetin plasma protein binding was determined by an ultrafiltration technique using an in vivo pharmacokinetic approach. Plasma protein binding data for the 3 doses studies in control animals could be described considering a single binding site with Kd = 21.9 +/- 2.1 microM and N = 0.98 +/- 0.05 sites per molecule of albumin. For hypoalbuminemic rats Kd was significantly increased (p < 0.05), while there was no significant change in the number of binding site per albumin molecule (Kd = 131.6 +/- 38.1 microM and N = 1.58 +/- 0.77). Our results show that hypoalbuminemia produces a disproportionate increase in the free fraction of tolmetin, not only by reducing albumin concentration, but also by a decrease in affinity. The mechanism responsible of such changes in affinity remains to be elucidated. PMID- 12088762 TI - Synthesis and opioid activity of new fentanyl analogs. AB - Three new fentanyl analogs (compounds 3-4-5) have been synthesized and evaluated for antinociceptive properties using the writhing test. The analgesic property of the active compound, N-[1-phenylpyrazol-3-yl]-N-[1-(2-phenethyl)-4-piperidyl)] propenamide (compound 4), was tested using the hot plate test in mice. Its opioid agonistic activity was characterized using three isolated tissues: guinea pig ileum, mouse vas deferens, and rabbit vas deferens. Compound 4 was as effective as fentanyl or morphine and it showed less antinociceptive potency than fentanyl but it was more potent than morphine. The duration of the antinociception was similar to that of fentanyl. This compound inhibited the electrically evoked contractions of myenteric plexus-longitudinal muscle strips of guinea pig ileum and of mouse vas deferens but not those of rabbit vas deferens. These effects could be reversed by micro selective antagonists (naloxone and/or CTOP) but not by the delta selective antagonist naltrindole, thus indicating that the compound acted as a micro opioid agonist. Finally, the binding data confirmed that compound 4 had high affinity and selectivity for the micro-receptor. PMID- 12088763 TI - Excitatory action of lead on rat sympathetic preganglionic neurons in vitro and in vivo. AB - Lead exposure elicited an increase in blood pressure and was considered to be a cardiovascular risk factor. The involvements of sympathetic nervous system and circulating catecholamines have been implicated in lead-induced hypertension. This study examined the effects of PbCl(2) on sympathetic preganglionic neurons (SPNs) in vitro and in vivo. In vitro electrophysiological study showed that superfusion of a low concentration (5 microM) of PbCl(2), which had no effects on membrane potential and spontaneous discharge rate, enhanced excitatory postsynaptic potentials (EPSPs) in some of the SPNs examined but inhibited inhibitory postsynaptic potentials (IPSPs) in other SPNs tested. A higher concentration (50 microM) of PbCl(2) inhibited both EPSPs and IPSPs in all SPNs examined. In vivo study showed that intrathecal injection of PbCl(2) (10 and 100 nmol) via an implanted cannula to the T7-T9 segments of urethane-anesthetized rats increased both the heart rate and mean arterial pressure. The pressor and tachycardic responses of intrathecal PbCl(2) (100 nmol) were attenuated by pretreatment with intravenous administration of hexamethonium (10 mg/kg) or intrathecal AP-5 (DL-2-amino-5-phosphonovaleric acid, 100 nmol), but were not significantly antagonized by prior intrathecal administration of CNQX (6-cyano-7 nitroquinoxaline-2,3-dione, 100 nmol). Taken together, these results demonstrated that lead may exert a stimulatory effect on SPNs, which may result from the enhancement of EPSPs and inhibition of IPSPs by low concentrations of lead. PMID- 12088764 TI - Music, immunity and cancer. AB - The effects of music on the immune system and cancer development were evaluated in rodents subjected to sound stress. Animals were exposed daily to broad band noise around midnight and/or music for 5 hours on the following morning. Thymus and spleen cellularity, peripheral T lymphocyte population, the proliferative response of spleen cells to mitogen concanavalin A and natural killer cell activity were calculated in BALB/c mice. Sprague Dawley rats were injected i.v. with Walker 256 carcinosarcoma cells; 8 days later the rats were sacrificed and the number of metastatic nodules on the surface of the lungs was calculated macroscopically. Music reduced the suppressive effects of stress on immune parameters in mice and decreased the enhancing effects of stress on the development of lung metastases provoked by carcinosarcoma cells. Music enhanced the immune parameters and the anti-tumor response in unstressed rodents. Our data at present demonstrates that music can effectively reverse adverse effects of stress on the number and capacities of lymphocytes that are required for an optimal immunological response against cancer in rodents. PMID- 12088765 TI - Evidence for a deficient pancreatic beta-cell response in a rat model of hyperthyroidism. AB - To clarify mechanism behind the abnormal glucose tolerance, observed in hyperthyroidism, we studied genomic and nongenomic effects of thyroid hormone on insulin secretion using a rat model of hyperthyroidism. Male Sprague-Dawley rats were intraperitoneally injected with vehicle, low (100 microg/kg) or high dose (600 microg/kg) of thyroxin (T(4)) for 2 weeks. Rats treated with high dose, but not low dose, of T(4), showed an increase in serum T(3) levels, and a decrease in body weight as compared to control rats. In rats treated with either dose of T(4), fasting blood glucose levels were increased, but serum insulin levels were similar to those of controls. After an oral glucose load, blood glucose levels were increased in rats treated with high dose, but not low dose, of T(4). Serum insulin levels after the oral glucose load were decreased in rats treated with either dose of T(4). After an intravenous glucose load, blood glucose levels were comparable among groups, but serum insulin levels tended to be low in T(4) treated rats. Steady-state blood glucose levels were comparable among groups. The insulin secretory responses to high glucose (20mM) or arginine (10mM) of the isolated pancreas was decreased in rats treated with high dose, but not low dose, of T(4). Mean insulin secretory response to glucose and arginine were decreased by 40.1% and by 60.4% in high-dose-T(4)-treated rats. Addition of T(3) in the perfusion medium decreased glucose-induced insulin release. Ratios of proinsulin mRNA levels to beta-actin mRNA were decreased in the islets of T(4)-treated rats (0.45 +/- 0.07 vs control 0.61 +/- 0.03, p < 0.05). Levels of TR (thyroid hormone nuclear receptor) alpha1 + cErb Aalpha2 mRNA, but not TRbeta1, were decreased in the pancreatic islets of T(4)-treated rats. Calculated islet area was increased, but the number of beta-cells determined immunohistochemically was not increased in T(4)-treated rats, nor the volume density of insulin positive islets. We concluded that a deficient pancreatic beta-cell response to glucose, rather than insulin resistance, was responsible for abnormal glucose tolerance in this model of hyperthyroidism. Thyroid hormone causes a decrease in glucose-induced insulin secretion. We observed nongenomic and genomic effects of thyroid hormone on glucose-induced insulin secretion. PMID- 12088766 TI - The anti-anginal drug fendiline elevates cytosolic Ca(2+) in rabbit corneal epithelial cells. AB - The effect of the anti-anginal drug fendiline on intracellular free Ca(2+) levels ([Ca(2+)](i)) in a rabbit corneal epithelial cell line (SIRC) was explored using fura-2 as a fluorescent Ca(2+) indicator. At a concentration above 1 microM, fendiline increased [Ca(2+)](i) in a concentration-dependent manner with an EC(50) value of 7 microM. The [Ca(2+)](i) response consisted of an immediate rise and an elevated phase. Extracellular Ca(2+) removal decreased half of the [Ca(2+)](i )signal. Fendiline induced quench of fura-2 fluorescence by Mn(2+) (50 microM), suggesting the presence of Ca(2+) influx across the plasma membrane. This Ca(2+) influx was abolished by La(3+) (50 microM), but was insensitive to dihydropyridines, verapamil and diltiazem. Fendiline (10 microM)-induced store Ca(2+) release was largely reduced by pretreatment with thapsigargin (1 microM) (an endoplasmic reticulum Ca(2+) pump inhibitor) to deplete the endoplasmic reticulum Ca(2+). Conversely, pretreatment with 10 microM fendiline abolished thapsigargin-induced Ca(2+) release. Fendiline (10 microM)-induced Ca(2+) release was not altered by inhibiting phospholipase C with 2 microM 1-(6-((17beta-3 methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122). Cumulatively, this study shows that fendiline induced concentration-dependent [Ca(2+)](i )increases in corneal epithelial cells by releasing the endoplasmic reticulum Ca(2+) in a phospholipase C-independent manner, and by causing Ca(2+) influx. PMID- 12088767 TI - Effect of timosaponin A-III, from Anemarrhenae asphodeloides Bunge (Liliaceae), on calcium mobilization in vascular endothelial and smooth muscle cells and on vascular tension. AB - The effects of timosaponin A-III (TA-III), from Rhizoma Anemarrhenae, on Ca(2+) mobilization in vascular endothelial cells and smooth muscle cells and on vascular tension have been explored. TA-III increased intracellular Ca(2+) concentrations ([Ca(2+)](i)) in endothelials cells at a concentration larger than 5 microM with an EC(50) of 15 microM, and increased [Ca(2+)](i) in smooth muscle cells at a concentration larger than 1 microM with an EC(50) of 8 microM. Within 5 min, the [Ca(2+)](i) signal was composed of a gradual rise, and the speed of rising depended on the concentration of TA-III. The [Ca(2+)](i) signal was abolished by removing extracellular Ca(2+) and was recovered after reintroduction of Ca(2+). The TA-III-induced [Ca(2+)](i) increases in smooth muscle cells were partly inhibited by 10 microM nifedipine or 50 microM La(3+), but was insensitive to 10 microM verapamil and diltiazem. TA-III (10-100 microM) inhibited 0.3 microM phenylephrine-induced vascular contraction, which was abolished by pretreatment with 100 microM N(omega)-nitro-L-arginine (L-NNA) or by denuding the aorta. TA III also increased [Ca(2+)](i) in renal tubular cells with an EC(50) of 8 microM. Collectively, the results show for the first time that TA-III causes [Ca(2+)](i) increases in the vascular system. TA-III acted by causing Ca(2+) influx without releasing intracellular Ca(2+). TA-III induced relaxation of phenylephrine induced vascular contraction via inducing release of nitric oxide from endothelial cells. PMID- 12088768 TI - A synthetic analog of prostaglandin E(1) prevents the production of reactive oxygen species in the intestinal mucosa of methotrexate-treated rats. AB - Administration of methotrexate to rats results in severe enterocolitis and death. Previous our studies showed that a synthetic analog of prostaglandin E(1), OP 1206 [17S, 20-dimethyl-trans-Delta(2)-prostaglandin E(1)] ameliorated the anticancer agent-induced enterocolitis of rats. In the current study, we have focused on the biochemical effect of OP-1206 on the methotrexate-induced intestinal inflammation implicating reactive oxygen species (ROS). Methotrexate (15 mg/kg body weight) was orally administered to rats once daily for 5 days. OP 1206 (0.5 microg/kg body weight) was orally administered to rats twice a day for 5 days. On the 6th day, the chemiluminescence from the jejunum was measured to evaluate the generation of ROS. Spontaneous chemiluminescence from the jejunum of the methotrexate-treated rats increased significantly, compared with the control. Luminol-enhanced chemiluminescence from inflamed mucosal scrapings from the jejunum of the methotrexate-treated rats indicated more remarkable enhancement than the control rats. The treatment of OP-1206 with methotrexate showed significantly lower chemiluminescence of both the jejunum and mucosal scrapings than those of the methotrexate-treated rats. The alkaline phosphatase (ALP) activity, as a marker of small intestinal differentiation, in the intestinal mucosa of the methotrexate-treated rats decreased remarkably, but that of the methotrexate and OP-1206-treated rats was significantly higher than that of the methotrexate-treated rats. Thus, OP-1206 may possibly help the anticancer chemotherapy by protecting the small intestine from the methotrexate-induced damage. PMID- 12088769 TI - A risk model derived from the National Registry of Myocardial Infarction 2 database for predicting mortality after coronary artery bypass grafting during acute myocardial infarction. AB - The mortality risk associated with coronary artery bypass grafting (CABG) after acute myocardial infarction (AMI) remains controversial. Although elective CABG is quite safe, the effects of recent myocardial infarction, gender, and other clinical factors on perioperative mortality rates are not completely understood. The objective of this study was to determine in-hospital mortality rates for patients with AMI receiving CABG and to generate a model to predict the risk for any individual patient with specific risk factors. Using the National Registry of Myocardial Infarction 2 database, we identified 71,774 subjects (21,270 women) with AMI who underwent CABG; we excluded those subjects who received immediate surgery as reperfusion therapy. Multivariate logistic regression was used to quantify the independent effects of age, recent myocardial infarction, gender, and other covariates on mortality. A risk score was then generated from the regression model to quantify the mortality risk. The results of logistic regression modeling determined that age was an independent predictor of in hospital death (adjusted odds ratio [OR] 3.05, 95% confidence interval [CI] 2.76 to 3.37 for age >75), as were previous CABG (OR 2.84, 95% CI 2.55 to 3.16), heart failure on presentation (OR 1.73, 95% CI 1.57 to 1.91 for Killip class II), and female gender (OR 1.58, 95% CI 1.45 to 1.71). The mortality risk score showed that 55% of patients had risk scores of 2 to 5 and mortality rates of 4% to 13%. This moderate risk group experienced 76% of the total predicted mortality. Thus, in-hospital CABG mortality rates after AMI are high compared with elective surgery. Using the described risk score, clinicians can quantify the impact of patient risk factors in making decisions about referral for and timing of CABG. PMID- 12088770 TI - Impact of spontaneous anterograde flow of the infarct artery on left ventricular function in patients with a first anterior wall acute myocardial infarction. AB - To assess the impact of spontaneous anterograde flow of the infarct artery on outcomes in patients with acute myocardial infarction (AMI), we studied 478 patients with a first anterior wall AMI who underwent coronary angiography within 12 hours after the onset of chest pain; Thrombolysis In Myocardial Infarction (TIMI) 3 flow was obtained after reperfusion therapy. Patients were divided into 3 groups: 119 patients with spontaneous anterograde flow (initial TIMI 2 or 3 flow) of the infarct artery, 118 patients with an initially occluded artery (TIMI 0 or 1 flow) and time to angiography or=55% (odds ratio 7.13, 95% confidence interval 3.10 to 16.4, p <0.001). In conclusion, although very early reperfusion improved LV function more than late reperfusion, spontaneous anterograde flow was associated with better acute and predischarge LV function after AMI compared with very early reperfusion of an initially occluded artery. PMID- 12088771 TI - Assessment by radionuclide ventriculography of postischemic regional left ventricular dysfunction in patients with healed myocardial infarction or angina pectoris. AB - Postischemic left ventricular (LV) dysfunction has been observed in experimental animal models after brief, complete coronary artery occlusion followed by reperfusion, but less relevant information is available for humans. The incidence and duration of postischemic LV dysfunction was examined by exercise radionuclide ventriculography in 50 patients with coronary artery disease. Cardiac imaging was performed at rest and was repeated during exercise and then immediately after and 5, 10, and 20 minutes after exercise. LV regional wall motion abnormalities were noted in 50 segments during exercise; they persisted in 30 of 50 segments after exercise, and remained apparent for 20 minutes in 11 segments. In contrast, in 33 segments, wall motion abnormalities were noted only after exercise and continued for 20 minutes in 13 of 33 segments. Exercise-related wall motion abnormalities were observed in 63 segments (76%) after exercise, and in 24 of 63 segments abnormalities continued for 20 minutes after exercise, although parameters of LV hemodynamic functions approached normal values after exercise. The mechanism of postexercise dysfunction is considered to involve acute myocardial stunning after a brief episode of myocardial ischemia, whereas regional wall motion abnormalities observed only after exercise seem to be related to increased levels of catecholamines or sympathetic overdrive, which mask less significant myocardial ischemia during exercise. PMID- 12088772 TI - Contribution of abdominal obesity and hypertriglyceridemia to impaired fasting glucose and coronary artery disease. AB - Multiple logistic regression models were used in a cross-sectional study to determine the relation of fasting glycemia to angiographically assessed coronary artery disease (CAD) in 569 men (aged 18 to 69 years) who were stratified according to fasting blood glucose concentrations (<6.1 mmol/L, and 6.1 to 6.9 mmol/L or 110 to 124 mg/dl), waist circumference (<90 vs >or=90 cm), and fasting triglyceridemia (<2.0 vs >or=2.0 mmol/L or <177 vs >or=177 mg/dl). For this purpose, nondiabetic impaired fasting glucose was defined as from 6.1 to 6.9 mmol/L (110 to 124 mg/dl) compared with 250 normoglycemic controls (fasting glycemia <6.1 mmol/L or <124 mg/dl) without history of CAD. In the absence of "hypertriglyceridemic waist," impaired fasting glucose was not predictive of CAD. However, the risk of CAD was markedly higher among subjects characterized by both the hypertriglyceridemic waist phenotype and the presence of impaired fasting glucose (odds ratio 8.5, 95% confidence intervals 3.5 to 20.4; p <0.05) compared with the normoglycemic group with low waist circumferences and triglyceride levels. Thus, the results of the present study emphasizes the importance of other underlying metabolic abnormalities, such as abdominal obesity and related atherogenic dyslipidemia, in the modulation of the CAD risk associated with hyperglycemia. PMID- 12088773 TI - Discrimination of early/intermediate and advanced/complicated coronary plaque types by radiofrequency intravascular ultrasound analysis. AB - Radiofrequency intravascular ultrasound (IVUS-RF) analysis, as an extension of conventional IVUS imaging, may provide more accurate plaque discrimination. Thirty-two autopsy atherosclerotic coronary arteries were investigated. Corresponding sectors in different plaques were matched by histologic and RF analysis. Histologic analysis utilized the American Heart Association plaque classification. The backscattered ultrasound RF signal was analyzed by fast Fourier transform, providing the underlying frequency components of its power spectrum. The normalized backscattered signal power (in decibels [dB]) for frequencies between 15.3 and 40.3 MHz was then measured for plaque discrimination. Advanced/complicated plaque types showed a higher signal power at all frequencies than early/intermediate lesion types (p <0.001 to p = 0.005). Discrimination of advanced/complicated lesion types was best at 15.3 MHz, with a cut-off point of 2.5 dB (sensitivity 93%, specificity 79%), and second best at 17.6 MHz (sensitivity 87%, specificity 71%, cut-off point 1.9 dB). With conventional IVUS, plaque discrimination was weaker; the best sensitivity for diagnosing early/intermediate lesion types was reached for "soft plaque" (sensitivity 63%, specificity 73%). Compared with conventional IVUS, IVUS-RF can discriminate between advanced/complicated and early/intermediate coronary atherosclerotic lesions with relatively high sensitivity and specificity in vitro. PMID- 12088774 TI - Prognostic value of heart rate variability in chronic congestive heart failure (Veterans Affairs' Survival Trial of Antiarrhythmic Therapy in Congestive Heart Failure). AB - Although the value of heart rate variability (HRV) for risk stratification after acute myocardial infarction has been demonstrated, the value of low HRV as a predictor of sudden cardiac death in patients with ischemic cardiomyopathy has not been shown convincingly to date. We retrospectively analyzed electrocardiographic data from 179 patients in the Veterans Affairs' Survival Trial of Antiarrhythmic Therapy in Congestive Heart Failure to determine if HRV (expressed as the SD of the normal-to-normal RR intervals [SDNN]) would be useful as a predictor of overall mortality and sudden death. Because our goal was to identify high-risk patients, we compared patients in the lowest quartile of HRV with the remaining patients. Among the 127 patients meeting inclusion criteria, SDNN <65.3 ms (the lowest quartile) was the sole independent factor predictive of survival in a multivariate model (p = 0.0001). A Cox proportional-hazards model revealed that each increase of 10 ms in SDNN conferred a 20% decrease in risk of mortality (p = 0.0001). Furthermore, patients with SDNN <65.3 ms had a significantly increased risk of sudden death (p = 0.016). Thus, HRV was the sole independent predictor of overall mortality and was significantly associated with sudden death in this population. PMID- 12088775 TI - Comparison of interstudy reproducibility of cardiovascular magnetic resonance with two-dimensional echocardiography in normal subjects and in patients with heart failure or left ventricular hypertrophy. AB - Fast breath-hold cardiovascular magnetic resonance (CMR) shows excellent results for interstudy reproducibility of left ventricular (LV) volumes, ejection fraction, and mass, which are thought to be superior to results of 2-dimensional echocardiography. However, there is no direct comparison of the interstudy reproducibility of both methods in the same subjects. A total of 60 subjects (normal volunteers [n = 20], or patients with heart failure [n = 20] or LV hypertrophy [n = 20]) underwent 2 CMRs and 2 echocardiographic studies for assessment of LV volumes, function, and mass. The interstudy reproducibility coefficient of variability was superior for CMR in all groups for all parameters. Statistical significance was reached for end-systolic volume (4.4% to 9.2% vs 13.7% to 20.3%, p <0.001), ejection fraction (2.4% to 7.3% vs 8.6% to 19.4%, p <0.001), and mass (2.8% to 4.8% vs 11.6% to 15.7% p <0.001), with a trend for end diastolic volume (2.9% to 4.9% vs 5.5% to 10.5%, p = 0.17). The superior interstudy reproducibility resulted in considerably lower calculated sample sizes (reductions of 55% to 93%) required by CMR compared with echocardiography to show clinically relevant changes in LV dimensions and function. Thus, CMR has excellent interstudy reproducibility in normal, dilated, and hypertrophic hearts, and is superior to 2-dimensional echocardiography. PMID- 12088776 TI - Is it time to reassess the optimal timing of coronary artery bypass graft surgery following acute myocardial infarction? PMID- 12088777 TI - Aspirin beyond platelet inhibition. PMID- 12088778 TI - Impact of body mass index on outcome after percutaneous coronary intervention (the obesity paradox). PMID- 12088779 TI - Relation of amounts of narrowing to the length of the right coronary artery. PMID- 12088780 TI - Variation in the risk of onset of acute myocardial infarction during the menstrual cycle. PMID- 12088781 TI - Comparison of errors in estimating weight and in dosing of single-bolus tenecteplase with tissue plasminogen activator (TIMI 10B and ASSENT I). PMID- 12088782 TI - Relation of initial platelet counts to Thrombolysis In Myocardial Infarction-3 flow rates at 90 minutes after commencing fibrinolytic therapy in patients with acute myocardial infarction. PMID- 12088783 TI - Comparison of coronary imaging between magnetic resonance imaging and electron beam computed tomography. PMID- 12088784 TI - Outcome of treatment of aorto-ostial lesions involving the right coronary artery or a saphenous vein graft with a polytetrafluoroethylene-covered stent. PMID- 12088785 TI - Factor V Leiden, prothrombin G20210A, and risk of sudden coronary death in apparently healthy persons. PMID- 12088786 TI - Relation of degree of laser debulking of in-stent restenosis as a predictor of restenosis rate. PMID- 12088789 TI - Comparison of patients with syncope with left ventricular dysfunction and negative electrophysiologic testing to cardiac arrest survivors and patients with syncope and preserved left ventricular function and impact of an implantable defibrillator. PMID- 12088788 TI - Diagnosing prosthetic mitral valve thrombosis and the effect of the type of prosthesis. PMID- 12088787 TI - Relation of peripheral flow-mediated vasodilatation and coronary arterial calcium in young patients with heterozygous familial hypercholesterolemia. PMID- 12088790 TI - Frequency and associations of symptomatic deterioration after dual-chamber defibrillator implantation in patients with ischemic or idiopathic dilated cardiomyopathy. PMID- 12088791 TI - Effect of hormone replacement therapy on carotid arterial compliance in healthy postmenopausal women. PMID- 12088792 TI - Effect of beta blockers on new congestive heart failure in older persons with healed myocardial infarcts. PMID- 12088793 TI - Neurobiology of psychosocial treatment of mental stress-induced myocardial ischemia in men. PMID- 12088794 TI - Helicobacter pylori infection, infectious burden, and coronary artery disease. PMID- 12088796 TI - C-reactive protein and coronary artery disease: influence of obesity, caloric restriction and weight loss. AB - C reactive protein (CRP) values in blood are a good indicator of the likelihood of acute coronary and cerebral events in both healthy subjects and patients with coronary artery disease. This indicates that atherosclerotic lesions rich in inflammatory cells and cytokines are more likely to produce acute events either through vasospasm and/or thrombosis and also can be readily detected through elevations in CRP when measured using a high sensitivity assay (hsCRP). However the arterial wall is only one potential source of cytokines which induce CRP production. Fat cells also produce cytokines, in particular IL-6 which induces the synthesis of CRP by the liver. Obesity, especially abdominal obesity, is associated with elevations of hsCRP. This may be of pathogenic significance as CRP stimulates the uptake of LDL by macrophages, induces complement activation which may cause cellular damage in the artery, and enhances monocyte production of tissue factor, thus enhancing the risk of thrombosis. Caloric restriction and weight loss lowers IL-6 and CRP levels and may beneficially suppress an immune response. Whether particular dietary macronutrients or micronutrients alter IL-6 or CRP is unknown but this issue is clearly becoming more important. PMID- 12088797 TI - Polyunsaturated dietary fats change the properties of calcium sparks in adult rat atrial myocytes. AB - This study investigated the effects of dietary omega-3 polyunsaturated fatty acids on calcium handling mechanisms in cardiac myocytes, with the hypothesis that this effect underlies some of the antiarrhythmic properties of these compounds. Adult male Sprague Dawley rats had their standard chow supplemented with either lard (57% saturated and 40% monounsaturated fat), canola oil (60% monounsaturated, 33% polyunsaturated) or fish oil (78% polyunsaturated). Isolated cardiac atrial myocytes from these animals were loaded with fluo-3AM and examined with laser scanning confocal microscopy. The dietary interventions resulted in considerable changes in the membrane phospholipid composition of cardiac cell membranes, particularly the ratio of n-6 to n-3 (2.17 with lard supplement and 1.28 with fish oil supplement). Calcium sparks in myocytes from rats which received saturated fat were significantly more prolonged than those from rats which received fish oil. (Lard = 105.4 +/- 18.9 ms; Fish oil = 43.5 +/- 4.7 ms: mean +/- s.e.m). The results for canola oil were intermediate (56.4 +/- 9.0 ms). The prolongation of the sparks in rats fed lard was primarily due to a higher proportion of sparks with long plateaus and/or slowed kinetics in this group. The frequency of sparks was not significantly different in cells from any group. We conclude that calcium handling mechanisms in rat atrial myocytes are affected by inclusion of different fats in the diet, correlated with changes in the cell membrane phospholipid composition, and speculate that this may underlie some of the antiarrhythmic properties of these dietary compounds. PMID- 12088798 TI - Oleic, linoleic and linolenic acids enhance receptor-mediated uptake of low density lipoproteins in Hep-G2 cells. AB - In the present study, the binding, internalization and degradation of low-density lipoprotein (LDL) was investigated in Hep-G2 cells treated with 18:0, 18:1, 18:2 and 18:3. In non-treated control cells, the surface binding (heparin-releasable) of 125I-LDL progressed in a saturable manner reaching equilibrium within 2 h, amounting 24.0 +/- 1.1, 29.5 +/- 1.3 and 31.4 +/- 2.8 (ng/mg cell protein) at 1, 2 and 4 h, respectively. The cells rapidly internalized 125I-LDL reaching a plateau at 2 h (72.4 +/- 6.3/1 h, 96.7 +/- 4.3/2 h and 100.8 +/- 4.6 ng/mg protein/4 h, respectively). The degradation of internalized LDL progressed slowly during the first hour of incubation reflecting the time required to an uptake and delivery of LDL to the cellular lysosomes. The levels of degraded LDL discharged into the medium then increased rapidly in a linear manner after the initial lag period, amounting 16.8 +/- 1.2, 51.8 +/- 7.0 and 118.2 +/- 5.7 ng/mg protein at 1, 2 and 4 h, respectively. The treatment of cells with of 1.0 mM of fatty acids for 4 h resulted in a significant increase in the surface binding of 125I-LDL compared to the control (34.9 +/- 3.0), but it was significantly lower in cells exposed to 18:0 (48.2 +/- 2.0) than to 18:1 (56.8 +/- 5.1), 18:2 (56.0 +/- 3.5) and 18:3 (57.8 +/- 6.0 ng/mg protein/4 h) (P < 0.05). The levels of degraded LDL in cells remained nearly the same regardless of fatty acid treatments, but degraded LDL levels in the medium were much higher in cells exposed to 18:1 (167.6 +/- 10.1), 18:2 (159.8 +/- 7.7) and 18:3 (165.1 +/- 14.7) than to 18:0 (142.1 +/- 8.4) and the control (121.2 +/- 3.4 ng/mg protein/4 h) (P < 0.05). The present finding that 18:1 is equally effective in enhancing the receptor-mediated LDL uptake and its degradation as those of 18:2 and 18:3 suggests that the major action of 18:1 in lowering LDL-cholesterol levels also involves an increased clearance of LDL via hepatic LDL-receptors. PMID- 12088799 TI - Fatty acid composition and conjugated linoleic acid content of different tissues in rats fed individual conjugated linoleic acid isomers given as triacylglycerols small star, filled. AB - HEAT TREATMENT OF VEGETABLE OILS GAVE RISE TO FOUR MAIN CONJUGATED LINOLEIC ACID (CLA) ISOMERS : the 9c,11t, 9t,11t, 10t,12c and 10t,12t. The diet of male Wistar rats was supplemented with 150 mg/day either 9c,11t-, 9t,11t-, 10t,12c- or 10t,12t CLA isomers for 6 days and their effects on lipid composition were investigated in liver, heart, skeletal muscle Gastrocnemius, kidneys, brain and adipose tissue. The incorporation of all isomers was low (< 1.4%) and the level was as follows : adipose tissue > Gastrocnemius > liver, kidneys > brain. The main changes in the overall lipid composition were observed in skeletal muscle (Gastrocnemius) and in heart and were associated with feeding the 10t,12c and 10t,12t isomers. The diet enriched in 10t,12t CLA decreased the total long chain polyunsaturated fatty acid proportion in Gastrocnemius (from 18.4% to 14.4%) and increased that of 20:4 n-6 in heart (from 16.9 to 19.3%). The diet enriched in 10t,12c CLA decreased the monounsaturated fatty acid proportion in Gastrocnemius (from 32.0 to 26.1%) and produced an effect similar to the 10t,12t in heart. By contrast, the 9c,11t and 9t,11t isomers did not affect fatty acid composition in all tissues and organs. We concluded that ingestion of 10t,12c and 10t,12t CLA present in oils and in CLA mixtures could change muscle lipid composition. PMID- 12088800 TI - Effects of an antioxidant-rich juice (sea buckthorn) on risk factors for coronary heart disease in humans. AB - There is increasing evidence to support the hypothesis that free radical-mediated oxidative processes contribute to atherogenesis. More recently the ability of antioxidant nutrients to affect cell response and gene expression has been reported in vitro, providing a novel mechanistic perspective for the biological activity of antioxidants. Sea buckthorn (Hippophae rhamnoides L.) is a rich source of antioxidants both aqueous and lipophilic, as well as polyunsaturated fatty acids. The objective of the study was to characterize the antioxidant profile of Sea buckthorn juice (SBJ) and to evaluate its effect on plasma lipids, LDL oxidation, platelet aggregation and plasma soluble cell adhesion protein concentration. Twenty healthy male volunteers were given either a placebo or SBJ for 8 weeks. Additional daily intakes of vitamin C, alpha-tocopherol, beta carotene and flavonoids through SBJ supplementation were 462, 3.2, 1.0 and 355 mg respectively. There were no significant changes in plasma total cholesterol, LDL C, platelet aggregation or plasma intercellular cell adhesion molecule 1 (ICAM-1) levels between treatment groups. Although not significant, a 20% and 17% increase in plasma HDL-C and triacylglycerol (TAG) concentrations were observed. SBJ supplementation also resulted in a moderate decrease in the susceptibility of LDL to oxidation. PMID- 12088801 TI - Involvement of DNA mismatch repair in folate deficiency-induced apoptosis small star, filled. AB - Folate is a critical factor for DNA metabolism and its deficiency is associated with a number of human diseases and cancers. Although it has been shown that folate deficiency induces genomic instability and apoptotic cell death, the underlying mechanism is largely unknown. Given the role of mismatch repair in maintaining genomic integrity, mismatch repair was tested for its involvement in folate deficiency-induced genomic instability and cell death. Cells proficient in mismatch repair were highly sensitive to folate deficiency compared with cells defective in either hMutSalpha or hMutLalpha. Since wild-type cells but not mutant cells underwent apoptosis upon extensive folate depletion, the apoptotic response is dependent on a functional mismatch repair system. Our data also indicate that p53 is required for the folate depletion-induced apoptosis. In vitro biochemical studies demonstrated that hMutSalpha specifically recognized DNA damage induced by folate deficiency, suggesting a direct participation of mismatch repair proteins in mediating the apoptotic response. We conclude that while the mismatch repair-dependent apoptosis is necessary to protect damaged cells from tumorigenesis, it may damage a whole tissue or organ, as seen in patients with megaloblastic anemia, during extensive folate deficiency. PMID- 12088802 TI - Influence of probiotic supplemented infant formula on composition of plasma lipids in atopic infants. AB - Probiotic therapy is a new, successful approach to alleviating allergic symptoms. In this study, our aim was to investigate whether the positive results obtained with probiotic therapy would be associated with the differential absorption and utilization of dietary PUFA. 15 infants referred to a pediatric clinic on the basis of atopic eczema were weaned to Bifidobacterium Bb-12 or Lactobacillus GG supplemented infant formula, or to the same formula without probiotics (randomized, placebo-controlled, double blind study design). In plasma neutral lipids, alpha-linolenic acid (18:3 n-3) proportions were reduced by the probiotic supplementation. In phospholipids, Lactobacillus GG supplemented formula did not influence alpha-linolenic acid proportions, while Bifidobacterium Bb-12 supplemented formula increased the proportion of alpha-linolenic acid; from 0.13 +/- 0.03 to 0.24 +/- 0.03 (mean +/- SEM) (P = 0.002). These results show that some physiological effects of probiotics may be associated with physiological interactions between probiotics and dietary PUFA. PMID- 12088803 TI - Common practice in molecular biology may introduce statistical bias and misleading biological interpretation. AB - In studies on enzyme activity or gene expression at the protein level, data are usually analyzed by using a standard curve after subtracting blank values. In most cases and for most techniques (spectrophotometric assays, ELISA), this approach satisfies the basic principles of linearity and specificity. In our experience, this might be also the case for Western-blot analysis. By contrast, mRNA data are usually presented as arbitrary units of the ratio of a target RNA over levels of a control RNA species. We here demonstrate by simple experiments and various examples that this data-normalization procedure may result in misleading conclusions. Common molecular biology techniques have never been carefully tested according to the basic principles of validation of quantitative techniques. We thus prefer a regression-based approach for quantifying mRNA levels relatively to a control RNA species by Northern-blot, semi-quantitative RT PCR or similar techniques. This type of techniques is also characterized by a lower reproducibility for repeated assays when compared to biochemical analyses. Therefore, we also recommend to design experiments, which allow the detection of a similar range of variance by biochemical and molecular biology techniques. Otherwise, spurious conclusions may be provided regarding the control level of gene expression. PMID- 12088804 TI - Relationship between clinical endpoints for induction of anesthesia and bispectral index and effect-site concentration values. AB - STUDY OBJECTIVE: To assess the relationship between clinical endpoints for induction of anesthesia and the electroencephalographic (EEG) bispectral index (BIS) and effect-site concentration (C(E)) values when using a target-controlled infusion (TCI) of either thiopental sodium or propofol, by hypothesizing that yawning may be a useful alternative to other commonly used clinical signs for determining loss of consciousness. DESIGN: Randomized observational clinical study. SETTING: Operating room of a university-based hospital. PATIENTS: 60 healthy adult patients (aged 20-50 yrs) scheduled for elective surgery with general anesthesia. INTERVENTIONS: During a TCI of propofol (n = 30) or thiopental (n = 30), clinical endpoints for loss of verbal responsiveness (LOV), loss-of-eyelash reflex (LOE), occurrence of yawning, and apnea were assessed at 15-second intervals. In addition, BIS and C(E) values were recorded at each of the endpoints. MEASUREMENTS AND MAIN RESULTS: In both anesthetic groups, the sequence of occurrence of the clinical endpoints was similar, namely LOV, LOE, yawning, and, lastly, apnea. Compared with LOV and LOE, yawning was associated with lower BIS and higher C(E) values with both anesthetics. The frequency of yawning was higher with thiopental than propofol (83% vs. 63%, respectively). However, the frequency of apnea was higher with propofol than thiopental (77% vs. 53%, respectively). CONCLUSION: The correlation of the clinical endpoints with BIS and C(E) values was highest for LOV. Yawning was as unreliable as LOE for determining the onset of unconsciousness during induction of anesthesia. This clinical sign failed to be observed in 17% and 37% of patients induced with thiopental and propofol, respectively. PMID- 12088805 TI - Effect of distance and infusion rate on operation of Medfusion 2010 infusion pump during magnetic resonance imaging. AB - STUDY OBJECTIVES: To determine the accuracy and reliability of intravenous infusion, as well as magnetic resonance image effect of the Medfusion 2010 infusion pump (Medex Medical Supplies, Inc., Duluth, GA) at distances of 2, 4, 8, 12, and 16 feet from a 1.5 Tesla MRI magnet over a four-week time interval, using infusion rates that would correspond to those of propofol administration. DESIGN: Prospective, open-label study. SETTING: Radiology department MRI in an American academic medical center. INTERVENTIONS: Five infusion pumps, including one outside the MRI suite as control, were tested. Pumps were evaluated at distances of 2, 4, 8, 12, and 16 feet from a 1.5 Tesla magnet. One pump at distances of 2 and 16 feet was tested during 30 and 90 hours of operation, respectively. Three pumps at distances of 4, 8, and 12 feet were tested during 120 hours of operation. Pump batteries were recharged for 18 hours outside the MRI suite between the 6-hour test periods. Distilled, deionized water was infused from a 30 mL or 60-mL syringe via a 36-inch Medex small-bore extension set into a graduated collection cylinder at rates of 5, 10.5, 21, or 42 mL/hr. Each rate was increased to the next infusion rate level at 30-hour weekly intervals. The collection cylinder with infused water was weighed on an electronic scale accurate to within +/-0.001 g. Analysis of variance and regression analysis were used to analyze data. A p < 0.05 value was considered statistically significant. MEASUREMENTS AND MAIN RESULTS: There was no significant difference comparing the grams weight of volume measured and the mL volume visualized within and between groups at varying distances from the magnet. Increasing infusion rates resulted in corresponding increases in volume delivered irrespective of the distance from the magnet. CONCLUSIONS: The Medfusion 2010 infusion pumps were found to be reliable and accurate, without causing failure or any significant degradation of MRI images compared to control at infusion rate of 42 mL/hr at a distance of 2 feet for 30 hours; and a rate of 5.0, 10.5, 21, and 42 mL/hr at 16 feet for 90 hours; and at 4, 8, and 12 feet for 120 hours of operation. PMID- 12088806 TI - Fluid management and postoperative respiratory disturbances in patients with transthoracic esophagectomy for carcinoma. AB - STUDY OBJECTIVE: To investigate whether intraoperative fluid management contributes to postoperative respiratory disturbances in esophagectomy for carcinoma. DESIGN: Retrospective study. SETTING: Operating room and postanesthetic care unit of the cancer center. PATIENTS: From 1997 to 2000, 112 ASA physical status I, II, and III patients with primary carcinoma of the esophagus undergoing transthoracic esophagectomy. INTERVENTIONS AND MEASUREMENTS: As of 1998, we altered fluid management during esophagectomy to save intraoperative fluid administration. Then, we investigated postoperative respiratory disturbances after esophagectomy in the period from 1998 to 2000 (late period) compared with the period from 1997 to 1998 (early period). We also investigated the relationship between perioperative risk factors and postoperative respiratory disturbances. The need for frequent (>10) bronchoscopic suctioning of sputum during postoperative period, the need for tracheostomy, and failure in the removal of endotracheal tube (ETT) (extubation) on the first postoperative day (1 POD) were investigated for respiratory disturbances after surgery. MAIN RESULTS: Intraoperative volume balance decreased more so in the late period compared with early period (p < 0.0,001). The need for tracheostomy, bronchoscopic suctioning, and extubation failure on 1 POD were more frequent in the early period than in the late period (p = 0.0083, p = 0.0319, and p = 0.0024, respectively). The hospital recovery period after surgery was shortened during the late period (p = 0.032). Intraoperative volume balance affected the need for tracheostomy and frequent bronchoscopy postoperatively. CONCLUSIONS: Careful intraoperative fluid administration may decrease postoperative respiratory disturbances. PMID- 12088807 TI - The effects of thiopental and generic and nongeneric propofol on respiratory resistance during anesthetic induction in patients with reactive airways. AB - STUDY OBJECTIVE: To demonstrate a favorable effect of propofol on respiratory system resistance during anesthetic induction, and to determine if generic propofol causes adverse effects on respiratory resistance. DESIGN: Randomized pilot study. SETTING: Anesthetic induction for elective surgery. PATIENTS: 27 consenting ASA physical status II and III patients with reactive airways (positive smoking history or chronic obstructive pulmonary disease), but not receiving bronchodilator therapy. INTERVENTIONS: Patients were randomized equally to one of three anesthetic induction (and maintenance) drugs: sodium thiopental, 5 mg/kg (25 microg/kg/min), generic or nongeneric propofol, 1.25 mg/kg (50 microg/kg/min). They received preinduction midazolam and fentanyl (2 mg and 150 microg) and intravenous lidocaine (0.5 mg/kg). After anesthetic induction, tracheal intubation was established, and predetermined settings for mechanical ventilation were initiated. MEASUREMENTS: Immediately after intubation, a sensor was placed on the 8-mm endotracheal tube to detect baseline airway pressure and flow. During maintenance, repeat measurements of pressure and flow were obtained at 2.5-minute intervals for 10 minutes. Respiratory system resistance was derived off-line using the isovolumetric technique. MAIN RESULTS: Patients were similar across groups. The respiratory resistance measured after anesthetic induction did not differ among groups. During the maintenance infusion of thiopental or propofol, respiratory resistance increased gradually across all groups. There was no difference in the response of respiratory resistance either at induction or during the 10-minute maintenance between the generic and the nongeneric propofol groups. CONCLUSIONS: In contrast to earlier reports, this pilot study was unable to document a difference in the respiratory resistance in patients induced with thiopental or propofol. In addition, we were unable to demonstrate any different respiratory responses between generic propofol, containing sodium metabisulfite preservative, and nongeneric propofol. PMID- 12088808 TI - Postoperative nausea and vomiting: comparison of the effect of postoperative meperidine or morphine in gynecologic surgery patients. AB - STUDY OBJECTIVE: To evaluate the incidence and severity of postoperative nausea and vomiting in women receiving postoperative intravenous morphine or meperidine following gynecologic surgery. DESIGN: Prospective, double-blind, randomized study. SETTING: Tertiary-care academic medical center. PATIENTS: 200 ASA physical status I, II, and III patients scheduled for elective gynecologic surgery. INTERVENTIONS: Patients received either postoperative IV morphine (n = 100) or meperidine (n = 100) following gynecologic surgery. MEASUREMENTS: We compared pain scores, sedation scores, nausea scores, well-being scores, vomiting rate, and patient satisfaction in both groups 15, 30, 60, and 120 minutes after arrival in the postoperative anesthesia care unit. MAIN RESULTS: The vomiting rate was 8/100 versus 7/100 (at 15 min), 4/100 versus 26/100 (at 30 min) (p < 0.05), 3/100 versus 23/100 (at 60 min) (p < 0.05), and 0/100 versus 0/100 (at 120 min) in the morphine or meperidine groups, respectively. The pain and sedation scores were similar in both groups. No major complications were noted in either group. CONCLUSION: Our study demonstrates an advantage of the use of morphine rather than meperidine for pain control in the immediate postoperative period following gynecologic surgery. PMID- 12088809 TI - The quotient end-tidal/inspired concentration of sevoflurane in a low-flow system. AB - STUDY OBJECTIVE: To investigate the effect of two different fresh gas flows on inspired and end-tidal sevoflurane concentration for a given vaporizer setting in a low-flow anesthesia system. DESIGN: Prospective clinical study. SETTING: Department of Anesthesiology of a university teaching hospital. PATIENTS: 56 ASA physical status I and II patients without systemic diseases, having elective surgery with an expected anesthesia time of at least 120 minutes. INTERVENTIONS: Patients were randomly assigned to receive either 1.0 or 2.0 L/min fresh gas flow with the vaporizer setting fixed at 2% sevoflurane. The inspired (In), end-tidal (Et), and Et/In ratio sevoflurane concentrations were estimated. MEASUREMENTS AND MAIN RESULTS: After 120 minutes of sevoflurane anesthesia the inspired and end tidal sevoflurane concentration were 1.45 +/- 0.10% versus 1.28 +/- 0.12% (p < 0.001) in the 1.0 L/min group and 1.64 +/- 0.08% versus 1.46 +/- 0.11% (p < 0.001) in the 2.0 L/min group. The ratio end-tidal and inspired concentrations/vaporizer setting was 0.64 +/- 0.06 and 0.73 +/- 0.05 in the 1.0 L/min group versus 0.73 +/- 0.05 and 0.82 +/- 0.04 in the 2.0 L/min group. For the ratio inspired and end-tidal/vaporizer setting there were significant difference between the groups (p < 0.001). The estimated ratio end-tidal/inspired was 0.88 +/- 0.04 in the 1.0 L/min group versus 0.89 +/- 0.04 in the 2.0 L/min group (ns). CONCLUSION: After 120 minutes of sevoflurane anesthesia at a vaporizer setting of 2% there is a significant difference between fresh gas flow of 1.0 and 2.0 L/min for inspired and end-tidal concentrations, but not for the ratio end-tidal/inspired. PMID- 12088810 TI - Airway length in adults: estimation of the optimal endotracheal tube length for orotracheal intubation. AB - STUDY OBJECTIVE: To estimate the optimal endotracheal tube (ETT) length in orotracheally intubated patients. DESIGN: Prospective study. SETTING: Operating room of a medical center hospital. PATIENTS: 293 ASA physical status I and II patients (150 male and 143 female), requiring general anesthesia and orotracheal intubation. INTERVENTIONS: We used fiberoptic bronchoscope within the ETT to identify the carina and vocal cords. MEASUREMENTS: The length from carina to vocal cords, vocal cords to right mouth angle (corner), and carina to right mouth angle were measured. The optimal ETT tip was defined as 5 cm above the carina. Patient's height and sternum length were recorded. MAIN RESULTS: The correlation between airway length and body height was significant. By linear regression, a formula was obtained to estimate the optimal ETT length in orotracheally intubated patients: the length from 5 cm above carina to right mouth angle (cm) =< body height (cm)/5> - 13. CONCLUSION: The optimal insertion length of the ETT for orotracheally intubated adult patients with the head placed in a neutral position is correlated with body height. The proposed formula can provide a useful guide to determine the optimal ETT tip position in most of the patients who required orotracheal intubation. PMID- 12088811 TI - Postoperative nausea and vomiting after total intravenous anesthesia with propofol and remifentanil or alfentanil: how important is the opioid? AB - STUDY OBJECTIVE: To compare the frequency and duration of postoperative nausea and vomiting (PONV) following total intravenous anesthesia (TIVA) with propofol and either remifentanil or alfentanil in outpatients undergoing arthroscopic surgery of the extremities. DESIGN: Randomized, third-party blinded study. SETTING: University medical center. PATIENTS: 100 ASA physical status I and II patients scheduled for arthroscopic surgery of the knee or shoulder. INTERVENTIONS: The anesthesia regimen consisted of a bolus followed by continuous infusion of propofol (2 mg/kg followed by 120 microg/kg/min) and the opioid (remifentanil 0.5 microg/kg followed by 0.1 microg/kg/min or alfentanil 10 microg/kg followed by 0.25 microg/kg/min). Patients breathed 100% oxygen spontaneously through a Laryngeal Mask Airway (or an endotracheal tube when medically indicated). Opioids were titrated to maintain blood pressure and heart rate within 20% of baseline and a respiratory rate of 10 to 16 breaths/min. Propofol was titrated downward as low as possible without permitting patient movement. MEASUREMENTS: Nausea was determined by an 11-point categorical scale and was recorded before surgery and multiple time points thereafter. The times of emetic episodes were recorded. Treatment of PONV was at the discretion of the postanesthesia care unit (PACU) nurses who were blinded to the identity of the opioid used. MAIN RESULTS: Nausea scores were 0 at all time points in over 70% of the patients in each group. None of the 100 patients vomited while in the hospital, and only one patient required antiemetic therapy. CONCLUSION: When propofol-based TIVA is used for arthroscopic surgery, short-acting opioids do not significantly affect the risk of PONV. PMID- 12088812 TI - Postoperative nausea and vomiting after craniotomy for tumor surgery: a comparison between awake craniotomy and general anesthesia. AB - STUDY OBJECTIVE: To assess the frequency of postoperative nausea and vomiting (PONV) in patients following an awake craniotomy compared to general anesthesia for tumor surgery. DESIGN: Prospective observational and chart review of all patients having a craniotomy for tumor during one year. SETTING: Postanesthesia care unit (PACU) and intensive care unit (ICU) of a university hospital. PATIENTS: 187 patients were reviewed. 107 patients who had a craniotomy for supratentorial tumor that was less than six hours in duration were analyzed and compared (50 awake craniotomy vs. 57 general anesthesia). INTERVENTIONS: Medical records were reviewed for events after the first four hours until discharge. The occurrence and the time of any nausea, vomiting; the administration of antiemetics and analgesic drugs; and complications were documented. MEASUREMENTS: Frequency of nausea, vomiting, administration of antiemetics and analgesia, and outcome between the two groups were compared using Chi-square and Student's t test. MAIN RESULTS: The frequency of nausea (4% vs. 23%; p = 0.012) and vomiting (0% vs. 11%; p = 0.052) were less in patients having an awake craniotomy compared to general anesthesia, but only during the first four hours. The administration of postoperative analgesia was not different between the two groups and did not influence the frequency of PONV. CONCLUSION: The frequency of PONV during the initial recovery phase was less in patients having an awake craniotomy for tumor surgery than in patients having a similar procedure with general anesthesia. PMID- 12088813 TI - Attitudes of anesthesiologists about addiction and its treatment: a survey of Illinois and Wisconsin members of the American Society of Anesthesiologists. AB - STUDY OBJECTIVE: To survey practicing anesthesiologists about their attitudes about addiction and its treatment by means of a previously validated instrument. DESIGN: Anonymous mail survey. SETTING: Metropolitan medical college. PARTICIPANTS: Active members of the American Society of Anesthesiologists in Illinois and Wisconsin. MAIN RESULTS: The survey consisted of 50 statements presented in a five item Likert (agreement-disagreement) format that evaluated five attitude factors: permissiveness, nonstereotypes, treatment interventions, treatment optimism, and nonmoralism. Five hundred twelve completed and 21 undeliverable surveys were returned from 1,656 surveys mailed (response rate = 31%). The raw scores for the five attitude factors were permissiveness 25 +/- 6, nonstereotypes 34 +/- 6, treatment interventions 32 +/- 4, treatment optimism 19 +/- 3, and nonmoralism 31 +/- 5 (means +/- SD). Anesthesiologists with a personal history of addiction recorded attitude scores that were significantly (p < 0.05) higher than those of their colleagues. Formal training in substance abuse management, attendance at a Twelve Step meeting as either a participant or an observer, and experience with a friend, relative, or colleague with addiction were also associated with significantly higher attitude scores. The attitude scores of anesthesiologists were consistently below those previously reported for clinicians who regularly care for patients with addiction. CONCLUSIONS: Personal experience with, or education about, addiction contributes to a more positive attitude about addiction. Anesthesiologists have less positive attitudes about addiction than do physicians who regularly manage the disease. PMID- 12088815 TI - Comparison of pulmonary arterial thermodilution and arterial pulse contour analysis: evaluation of a new algorithm. AB - STUDY OBJECTIVE: To compare cardiac index (CI) measurement by arterial pulse contour analysis using two different algorithms (CI(PC), CI(PCnew)) with pulmonary arterial thermodilution values (CI(PA)) so as to evaluate the difference between the conventional algorithm, CI(PC), and a new algorithm, CI(PCnew), that accounts for patients' individual aortic compliance. DESIGN: Prospective, clinical study. SETTING: Intensive care unit of a university hospital. PATIENTS: 20 ASA physical status II and III patients following elective cardiac surgery. MEASUREMENTS AND MAIN RESULTS: 360 parallel triplicate determinations of CI (CI(PA), CI(PC), CI(PCnew)) were performed within a 90 minute period during the immediate postoperative period. Prior to the start of the study period, CI(PC) as well as CI(PCnew) were calibrated by triplicate femoral arterial thermodilution measurements. Regression analysis of CI(PA) and CI(PC), as well as CI(PA) and CI(PCnew), revealed r = 0.89, p < 0.001, and r = 0.93, p < 0.001, respectively. Bland-Altman analysis was used for determining the accuracy and precision of CI(PC) and CI(PCnew) compared with CI(PA). The mean differences (m) and standard deviation (SD) between CI(PA) and CI(PC,) as well as CI(PA) and CI(PCnew), resulted in m = -0.312 L/min/m(2), SD = 0.456 L/min/m(2), and m = - 0.140 L/min/m(2), SD = 0.328 L/min/m(2), respectively. CONCLUSION: Arterial pulse contour analysis measurement of CI using either algorithm correlates well with CI values derived by pulmonary arterial thermodilution. However, the algorithm introduced in this study proved to be a more accurate predictor of values as derived by pulmonary artery catheter. PMID- 12088816 TI - Airway management of a patient with facial trauma. AB - Airway management in patients with facial trauma is usually challenging. In this case, we report the airway management of a patient who had multiple midface fractures and a two-inch tree limb implanted into his face. PMID- 12088814 TI - Rapid inhalation induction with 7% sevoflurane combined with intravenous midazolam. AB - STUDY OBJECTIVE: To identify an improved rapid inhalation induction (RII) technique by investigating hemodynamics and heart rate variability of induction with midazolam and RII with 7% sevoflurane in comparison with RII alone. DESIGN: Prospective, randomized study. SETTING: Operating room of a university hospitals. PATIENTS: 40 patients scheduled for gastrectomy without complications. INTERVENTIONS: In the sevoflurane (Sev) group, anesthesia was induced with tidal volume breathing of 7% sevoflurane with 50% nitrous oxide (N(2)O) in oxygen (O(2)) for 3 minutes. In the midazolam-sevoflurane group (Mid-Sev) intravenous midazolam 0.1 mg/kg was given, followed by 7% sevoflurane with 50% N(2)O. In both groups, endotracheal intubation was facilitated with vecuronium 0.15 mg/kg, which was administered at loss of consciousness. MEASUREMENTS: Blood pressure, heart rate, heart rate variability, time to loss of response to verbal command (sleep time), rate of body movement or cough during induction, and patient satisfaction were monitored. MAIN RESULTS: Blood pressure and heart rate increased significantly in the Sev group but not in the Mid-Sev group. Heart rate was significantly higher in the Sev group at 1 minute after intubation. High frequency component (cardiac parasympathetic activity) in heart rate variability increased before intubation, with a higher value seen in the Sev group than the Mid-Sev group. The ratio of low-frequency component to high-frequency component (cardiac sympathetic activity) did not change in either group. Sleep time did not differ between the two groups. The numbers of patients with body movement and cough, and patients who were not satisfied with the induction method were, respectively, four, three, and eight in the Sev group; and one, zero, and two in the Mid-Sev group. The last value was significantly higher in the Sev group (p = 0.0285). CONCLUSIONS: The addition of intravenous midazolam 0.1 mg/kg provides more stable hemodynamics, cardiac autonomic nervous system activity, and patient satisfaction in RII with 7% sevoflurane. PMID- 12088817 TI - Bispectral index monitoring during hypoglycemic coma. AB - Bispectral index (BIS) monitoring is gaining wide acceptance as a gauge of the hypnotic effect of anesthetics. However, change in BIS in relation to the mental status of diabetic patients due to hypoglycemia has not been reported to date. We describe a case in which BIS was found to correlate with a patient's level of consciousness and changes in blood glucose levels. PMID- 12088818 TI - Current status of the Combitube: a review of the literature. AB - The Combitube (Tyco-Healthcare-Kendall-Sheridan, Mansfield, MA) is an easily inserted and highly efficacious device to be used as an alternative airway whenever conventional ventilation fails. The Combitube allows ventilation and oxygenation whether the device locates in the esophagus (very common) or the trachea (rare). In this report, we review studies that suggest the Combitube is a valuable and effective airway in the emergency and prehospital settings, in cardiopulmonary resuscitation, in elective surgery, and in critically ill patients in the intensive care unit. Also reviewed are studies that demonstrate the superiority of the Combitube over other supraglottic ventilatory devices in resuscitation with respect to success rates with insertion and ventilation. Contrary to the Laryngeal Mask Airway, the Combitube may help in patients with limited mouth opening. The Combitube may be of special benefit in patients with massive bleeding or regurgitation, and it minimizes the risk of aspiration. PMID- 12088819 TI - Gaston Labat, John Lundy, Emery Rovenstine, and the Mayo Clinic: the spread of regional anesthesia in America between the World Wars. AB - The spread of regional anesthesia in America was greatly facilitated by the work of Gaston Labat. Recruited to work at the Mayo Clinic, Dr. Labat there published his seminal textbook, Regional Anesthesia, in which he laid out his techniques to the next generation of physician specialists, notably John Lundy, Ralph Waters, and Emery Rovenstine. It was Rovenstine who was responsible for creating the specialty of anesthesiology in the 1920s and 1930s. John Lundy continued Labat's work at the Mayo Clinic when Labat left for Bellevue Hospital in New York. There, while teaching, Labat further developed and refined his techniques for delivering regional anesthesia. PMID- 12088820 TI - Use of internal standard RNA molecules for the RT-PCR amplification of the faeces borne RNA viruses. AB - The diagnostic system based on reverse transcription (RT)-PCR has been used widely for the detection of viral genomes of faecal-borne RNA viruses. However, faecal specimens often produce both false positive and false negative results. Therefore, there is a need for a diagnosis procedure that can control for 'false results'. In this study, an internal standard RNA that can serve as a non competitive positive template was developed and used directly to detect faecal borne RNA viruses without noticeable competitive inhibition of the target viral genome. These results suggest that the internal standard RNA is a useful standard molecule when undertaking diagnostic qualitative RT-PCR procedures for enteroviruses and related faecal-borne RNA viruses. PMID- 12088821 TI - Differentiation of type A, Asia1 and O foot-and-mouth disease virus variants, amplified by the same system, by sequencing of the capsid protein genes. AB - A reverse transcription-dependent polymerase chain reaction (RT-PCR) is described that amplifies the genes encoding the capsid proteins VP1-3 of at least three evolutionary lineages each of the foot-and-mouth disease (FMD) virus types A, Asia1 and O. Most of these lineages are circulating at present in Asia and Africa. The method is not only suitable to confirm suspected outbreaks of FMD, but also describes the modulation of major and minor antigenic sites in the course of an epizootic by nucleotide sequence determination of the obtained RT PCR products. Such knowledge helps to choose suitable vaccines for disease control. PMID- 12088822 TI - An improved method for recovery of F-defective Sendai virus expressing foreign genes from cloned cDNA. AB - An improved system is described to recover non-transmissible Sendai virus that lack the envelope fusion (F) gene from cloned cDNA. The system (1) used plasmids that expressed the F and the HN viral envelope proteins, as well as the plasmids that expressed the viral NP, P, and L proteins as helper plasmids for recovery, and (2) overlaid packaging cells that expressed the F protein. With this improved system, we have succeeded in recovery of F-defective Sendai virus expressing two foreign proteins, and expression vectors that do not contain the EGFP reporter gene. This system may provide the basis for constructing recombinant F-defective Sendai virus for preventing and treating human diseases in the form of vaccines and vectors for gene therapy. PMID- 12088823 TI - Replication of rhesus cytomegalovirus in life-expanded rhesus fibroblasts expressing human telomerase. AB - The kinetics of rhesus cytomegalovirus (RhCMV) infection were compared in primary and telomerase-immortalized (Telo-) rhesus fibroblasts (RF). Equivalent viral titers were achieved with both cell types. However, the production of infectious virions was slightly faster and plaque size was larger in Telo-RF, compared with primary cells. Comparable RhCMV growth curves and viral susceptibility were observed using Telo-RF passaged for different time periods in culture, whereas the ability of primary cells to support robust RhCMV replication declined as the cells approached senescence. Analysis of cell growth kinetics suggested that the rate of RhCMV replication was directly related to the rate of cell proliferation. RT-PCR analysis of representative RhCMV genes demonstrated that the presence of telomerase did not alter the temporal profile of RhCMV gene expression. In addition, Telo-RF cells were observed to have a significantly increased efficiency of transfection with cationic lipids, compared with primary RF. These results demonstrated that Telo-RF represents a stable, permissive cell line for RhCMV infection, facilitating standardization of in vitro assays for this important non-human primate CMV. The ease of transfection will enable molecular analyses and the generation of complementing cell lines for the propagation of defective RhCMV variants. PMID- 12088824 TI - Inclusion of full length human immunodeficiency virus type 1 (HIV-1) gag sequences in viral recombinants applied to drug susceptibility phenotyping. AB - Drug susceptibility phenotyping of recombinant clinical human immunodeficiency virus type 1 (HIV-1) isolates has been used widely to quantitatively assess viral resistance to antiretroviral agents. A novel method is described for HIV-1 drug susceptibility phenotyping. Recombinant virus that contains the entire HIV-1 Gag, protease (PR) and reverse transcriptase (RT) coding regions is generated from plasma of HIV-1 infected subjects, thus allowing the in vitro investigation of effects caused by all protein-coding sequence elements upstream from the drug targets on: (i) drug susceptibility; and (ii) viral replicative capacity. Mutations known to cause retarded viral growth kinetics (RT M184V and PR I50V) were introduced and analyzed in parallel using both the new Five Prime HIV assay (FPH) and a standard recombinant virus assay (RVA). The M184V and I50V mutants produced up to 4.8- and 5.9-fold higher p24 antigen levels, respectively, with the FPH when compared to the cultures containing RVA-derived viruses. The reduced number of homologous recombination events necessary to generate replication competent provirus with the FPH is the most likely explanation for these findings. Long range RT-PCR products were generated from plasma of HIV-1 infected subjects and HIV-1 LTR sequences were added using one-step PCR-mediated recombination. FPH-recombinants generated from two patients with previous HIV PR and RT inhibitor therapy showed lower drug susceptibilities than mutants established in parallel by RVA, and relative in vitro replication of the FPH recombinant derived from one of these subjects was enhanced compared to the corresponding RVA mutant. Although there were changes from the HIV-1 subtype B consensus sequence in amino acids flanking the Gag p17/p24, p24/p2 or p2/p7 PR cleavage sites, none were within the 10 amino acids immediately flanking the sites. These data suggest that determinants of drug susceptibility may be encoded in Gag upstream of the p7/p1 and p1/p6 regions, and that some phenotyping assays may therefore be underdetermining the reduction of drug susceptibility in some viral isolates. PMID- 12088825 TI - Detection of reovirus by reverse transcription-polymerase chain reaction using primers corresponding to conserved regions of the viral L1 genome segment. AB - A rapid, reverse transcription-polymerase chain reaction (RT-PCR) procedure for the detection of reovirus RNA in cell culture is described. Total nucleic acids are extracted from a small volume of cell culture supernatant and reverse transcribed using random hexamers. An aliquot of cDNA is then utilized in nested PCR. The PCR primers correspond to sequences conserved between prototype reovirus strains type 1 Lang, type 2 Jones, and type 3 Dearing, as well as those of several reovirus field-isolate strains. Reactions are analyzed by agarose gel electrophoresis, and samples showing a band of the appropriate size in the first and second amplification, or in the second amplification alone, are designated as positive. This protocol allows for the rapid and sensitive detection of reovirus in cell culture. The RT-PCR methods described below can easily be adapted to the amplification of reovirus from other media, including preserved tissues, clinical specimens, and water. PMID- 12088826 TI - Purification and reactiongenicity of human immunodeficiency virus type 2 total external glycoprotein expressed in Pichia Pastoris. AB - The purification of human immunodeficiency virus type 2 total external glycoprotein gp105 expressed in Pichia pastoris was investigated. Expression conditions were optimized by an orthogonal test. The results from tests of variance analyses showed that the most important parameter for efficient expression of total gp105 in P. pastoris is adequate aeration during methanol induction. The optimum induction conditions for gp105 expression were: more than 85% aeration, induction for 3 days, the initial pH 6.0-7.0 and a final methanol concentration of 1.0%. Under these conditions, the expressed total gp105 was secreted into fermentation broth and reached a yield of 23%, approximately 141 mg/l. Expressed gp105 was isolated and purified by salting out and Sephadex G-100 chromatography and the yield of gp105 was 40%. gp105 was purified to electrophoretic purity and its isoelectric point (pI) was about 5.2 by SDS-PAGE and isoelectrofocusing. The purified gp105 contained approximately 35% carbohydrate, which proved that the expressed gp105 was a glycoprotein. Its N terminal amino acid was arginine by Dansyl-Cl and the result indicated that expressed gp105 was secreted and cleaved correctly. The results from gp105 ELISA demonstrated that the purified total gp105 showed good reactiongenicity and antigenic specificity. PMID- 12088827 TI - An improved endothelial barrier model to investigate dengue haemorrhagic fever. AB - A cell culture model suitable for studies of dengue haemorrhagic fever was developed, based on culture of primary human umbilical vein endothelial cells (HUVECs) on a permeable membrane. By electron microscopy, cultured HUVECs at day 11 resembled morphologically microvascular endothelium. Endothelial barrier function was assessed by measuring transendothelial flux of albumin. Instead of using a labelled tracer molecule, an enzyme-linked immunosorbent assay (ELISA) was developed to measure concentrations of native human albumin. The permeability characteristics of the HUVEC monolayer were found to be improved significantly (approximately 1 log reduction in permeability coefficient for albumin) by culturing HUVECs in human serum rather than fetal calf serum. Permeability coefficients for albumin in the range 1-4 x 10(-7) cm/s were achieved, which is an improvement on previous in vitro models of the endothelial barrier. Comparison of transendothelial flux of albumin and urea provided evidence of molecular sieving by the HUVEC monolayer. Moreover, tumour necrosis factor-alpha induced a dose-dependent, reversible increase in permeability of the HUVEC monolayer. This endothelial barrier model thus has many important characteristics that resembled human microvascular endothelium and is an improvement on the previous model proposed for studies of dengue haemorrhagic fever. PMID- 12088828 TI - Purification of turkey coronavirus by Sephacryl size-exclusion chromatography. AB - Sephacryl S-1000 size-exclusion chromatography was used to purify turkey coronavirus (TCoV) from infected turkey embryo. TCoV was propagated in the 22-day old turkey embryos. Intestines and intestinal contents of infected embryos were harvested and homogenized. After low speed centrifugation, the supernatant was concentrated by ultracentrifugation through a cushion of 30 or 60% sucrose solution, or by ammonium sulfate precipitation. The purification methods included sucrose gradient and Sephacryl S-1000 size-exclusion chromatography. Ultracentrifugation through a cushion of 60% sucrose solution was better than the other two methods for concentration of TCoV from intestinal homogenate. The most effective method for purifying TCoV and removing extraneous materials was size exclusion chromatography as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. More spike-rich particles were observed in the sample purified by chromatography than those purified by sucrose gradient as examined by electron microscopy. Differentiation of turkey anti-TCoV antiserum from normal turkey serum was better achieved by ELISA plates coated with TCoV preparation purified by size-exclusion chromatography than that purified by sucrose density gradient. The results indicated that Sephacryl S-1000 chromatography was useful for purification of TCoV. PMID- 12088829 TI - Development of a particle agglutination assay system for detecting Japanese encephalitis virus-specific human IgM, using hydroxyapatite-coated nylon beads. AB - Japanese encephalitis virus-specific IgM is a reliable indicator for serodiagnosis of Japanese encephalitis. A particle agglutination (PA) assay system was developed to detect anti-Japanese encephalitis virus IgM in human serum samples. The newly developed PA assay consisted of hydroxyapatite-coated nylon beads and V-bottom 96-well microplates. Hydroxyapatite-coated nylon beads were coated with Japanese encephalitis virus antigens. Japanese encephalitis virus antigen-coated, hydroxyapatite-coated nylon beads agglutinated in the IgM captured wells when anti-Japanese encephalitis virus IgM-positive serum samples were used. A button pattern was formed at the bottom of the wells when anti Japanese encephalitis virus IgM-negative serum samples were used. Thirty anti Japanese encephalitis virus IgM-positive serum samples from Japanese encephalitis confirmed cases were tested by the PA assay. All these serum samples were determined to be Japanese encephalitis virus IgM-positive. IgM titers determined by the PA assay corresponded to those determined by enzyme-linked immunosorbent assay. The titers were consistent in two independent PA assays. These results indicate that the newly developed PA assay is a reliable method for detecting anti-Japanese encephalitis virus IgM in human serum samples and that this assay will be a suitable diagnostic system especially in rural areas of Asia. PMID- 12088830 TI - Continuous porcine cell lines developed from alveolar macrophages: partial characterization and virus susceptibility. AB - Porcine monomyeloid cell lines were established following transfection of primary porcine alveolar macrophage cultures with plasmid pSV3neo, carrying genes for neomycin resistance and SV40 large T antigen. The parental clone 3D4 exhibited a relatively rapid doubling time (25.5 h), high plating efficiency and mixed phenotype with respect to growth on a solid support. Single cell cloning of the 3D4 parent resulted in establishment of several cell lines; three of them designated 3D4/2, 3D4/21 and 3D4/31 were selected for further characterization. All three clones supported the replication of vesicular stomatitis virus (VSV), pseudorabies virus (PRV), classical swine fever virus (CSFV), swine vesicular disease virus (SVDV), swine poxvirus, African swine fever virus (ASFV), herpes simplex virus (HSV), parainfluenza virus, bovine adenovirus (BAV), vaccinia virus (VV), and porcine adenovirus (PAV). Under the conditions tested the cells did not support replication of porcine reproductive and respiratory syndrome virus (PRRSV). The swine myeloid character was confirmed for all three clones by fluorescence activated cell scanning (FACS) analysis using monoclonal antibodies 74-22-15 and DH59B, which recognize the pan-myeloid antigen cluster SWC3a. A subpopulation of each cell line was positive for nonspecific esterase activity and phagocytic activity to varying degrees depending on the media formulation. Cells from all three lines exhibited anchorage dependent growth when maintained in RPMI 1640 medium supplemented with 5-15% fetal bovine serum (FBS) and nonessential amino acids. Propagation in commercially formulated serum free media resulted in colony formation and growth in suspension. The addition of dimethyl sulfoxide (DMSO) or phorbol 12-myristate 13-acetate (PMA) to serum free media restored cell attachment. DMSO was also able to induce expression of CD14 monocyte marker in the 3D4/31 cell line maintained in FBS containing medium, as determined by FACS with monoclonal antibody CAM36A. Supplementation of RPMI medium with 10% porcine serum upregulated the expression of CD14 and induced expression of porcine macrophage markers recognized by antibodies 2B10 and 2G6 (Vet. Immunol. Immunopathol. 74 (2000) 163) in all three cell lines. The porcine myelomonocytic cell lines obtained may have a wide variety of applications in porcine virology and immunology. PMID- 12088831 TI - Pretreatment to avoid positive RT-PCR results with inactivated viruses. AB - Enteric viruses that are important causes of human disease must often be detected by reverse transcription-polymerase chain reaction (RT-PCR), a method that commonly yields positive results with samples that contain only inactivated virus. This study was intended to develop a pretreatment for samples, so that inactivated viruses would not be detected by the RT-PCR procedure. Model viruses were human hepatitis A virus, vaccine poliovirus 1 and feline calicivirus as a surrogate for the Norwalk-like viruses. Each virus was inactivated (from an initial titer of approximately 10(3) PFU/ml) by ultraviolet light, hypochlorite or heating at 72 degrees C. Inactivated viruses, that were treated with proteinase K and ribonuclease for 30 min at 37 degrees C before RT-PCR, gave a negative result, which is to say that no amplicon was detected after the reaction was completed. This antecedent to the RT-PCR method may be applicable to other types of viruses, to viruses inactivated in other ways and to other molecular methods of virus detection. PMID- 12088832 TI - Cyclosporin A in blood and brain tissue following intra-carotid injections in normal and stroke-induced rats. AB - Administration of Cyclosporin A (CsA) to rats undergoing reversible global or focal ischemia has been demonstrated to be variably neuroprotective. As CsA does not readily cross the blood-brain barrier, the variability may be due to differences in bioavailability of CsA to the ischemic brain. We have, therefore, quantitated CsA levels in blood and brain following intra-carotid injection in rats undergoing permanent right middle cerebral artery (MCA) occlusion using a three-vessel model of focal cerebral ischemia. After 30 min of three-vessel occlusion, CsA (10 mg/kg) was injected into the left external carotid artery followed by reversal of the left common carotid artery occlusion. At various times post-injection, blood samples were collected from the vena cava and samples of ischemic or sham-operated cortex were obtained for CsA quantitation by tandem mass spectrometry. Pharmacokinetic parameters were determined using non-linear mixed-effects modeling. CsA areas under the curve between normal and stroke induced rats were not significantly different in blood (18355 vs. 19405 ng x h/ml, NS) or in brain tissue (15664 vs. 14931 ng x h/g, NS). These results demonstrate that intra-carotid injection of CsA results in high levels in brain (brain-blood ratio from 0.5 to 1). No significant differences in blood and brain exposure were observed between normal and stroke-induced rats. Therefore, reduced cerebral blood flow in the ischemic territory did not limit CsA availability to the cortex. In addition, CsA intra-carotid administration was neuroprotective following 24 h recovery as there was a significant decrease in the infarct area of the affected hemisphere compared to saline injected rats as estimated by TTC staining of viable tissue. PMID- 12088833 TI - Optic afferents to the parabrachial nucleus. AB - Following intraocular injection of cholera toxin subunit B (CTB), optic afferents to the dorsal pontine region were observed in Mongolian gerbils, Chilean degus, and laboratory rats. CTB-positive optic axons emerge at the caudal pole of the superior colliculus, descend through the periaqueductal gray, and innervate the lateral parabrachial nucleus. This projection appears to be a continuation of the retinal pathway that innervates the dorsal raphe nucleus in these same species. PMID- 12088834 TI - Regulation of interstitial excitatory amino acid concentrations after cortical contusion injury. AB - Increases in brain interstitial excitatory amino acid (EAA(I)) concentrations after ischemia are ameliorated by use-dependent Na+ channel antagonists and by supplementing interstitial glucose, but the regulation of EAA(I) after traumatic brain injury (TBI) is unknown. We studied the regulation of EAA(I) after TBI using the controlled cortical impact model in rats. To monitor changes in EAA(I), microdialysis probes were placed in the cortex adjacent to the contusion and in the ipsilateral hippocampus. Significant increases in dialysate EAA(I) after TBI were found compared to levels measured in sham controls. Treatment with the use dependent Na+ channel antagonist 619C89 (30 mg/kg i.v.) did not significantly decrease dialysate glutamate compared to vehicle controls in hippocampus (10.4+/ 2.4 vs. 11.9+/-1.6 microM), but there was significant decrease in dialysate glutamate in cortex after 619C89 treatment (19.3+/-3 vs. 12.6+/-1.1 microM, P<0.05). Addition of 30 mM glucose to the dialysate, a treatment that decreases EAA(I) after ischemia, had no significant effect upon dialysate glutamate after TBI in cortex (20.0+/-4.9 vs. 11.7+/-3.4 microM) or in hippocampus (10.9+/-2.0 vs. 8.9+/-2.4 microM). These results suggest that neither increased release of EAAs due to Na+ channel-mediated depolarization nor failure of glutamate reuptake due to glucose deprivation can explain the majority of the increase in EAA(I) following TBI. PMID- 12088835 TI - Stress-induced dynamic changes in mouse brain polyamines. Role in behavioral reactivity. AB - Recent findings indicate that rapid and transient changes in polyamine metabolism, termed the polyamine-stress-response, may occur repeatedly in the brain after chronic intermittent stress. Here, we sought to examine the effects of chronic intermittent restraint stress, or of daily intraperitoneal dexamethasone injections on polyamine concentrations in the hippocampus of adult male C57BL/6 mice. Additionally, we studied the effects of alpha difluoromethylornithine, an irreversible ornithine decarboxylase inhibitor, on stress-induced changes in polyamines and on behavioral reactivity to novelty stress measured in an open-field arena. As previously observed, following a single stress episode putrescine concentration increased transiently, but the polyamines spermidine and spermine remained unchanged. Following chronic restraint stress, putrescine concentration was increased after each daily stress episode with the largest increase observed after the 4th episode, while spermidine was increased just after the 2nd and 4th episodes and spermine only after the 4th daily episode. In contrast, all polyamine concentrations were increased after 10 injections of either dexamethasone or vehicle (0.9% NaCl). A 14-day course of alpha-difluoromethylornithine treatment resulted in a complete putrescine depletion and over 50% reduction in polyamines, and led to changes in open field activity indicative of altered emotional behavior. CONCLUSIONS: (a) while putrescine concentration increases in the hippocampus after each restraint stress episode, spermidine and spermine undergo a delayed but transient increase; (b) in contrast, chronic dexamethasone treatment may lead to a permanent increase in the concentrations of all polyamines and; (c) chronic alpha difluoromethylornithine treatment reduces brain polyamine concentrations and modulates emotional reactivity to novelty stress. The study indicates that the brain polyamine-stress-response is a dynamic process that varies with the type, intensity and length of stressful stimuli, and implicates this response as an adaptive mechanism in the reaction to stressors. PMID- 12088836 TI - Short therapeutic window for nifedipine in transient intrauterine ischemia in fetal rat brain. AB - The aim of this study was to explore whether nifedipine influences the secondary deterioration of cerebral mitochondrial function after transient intrauterine ischemia in fetal rats. Intrauterine ischemia was induced by a 30-min occlusion of the right uterine artery at 20 days of gestation in Wistar rats. Nifedipine (1 mg kg(-1)) or vehicle was injected subcutaneously before the onset of ischemia or 1 h after the start of recirculation. Fetuses were delivered by cesarean section at the end of ischemia (n=6 with vehicle; n=6 with nifedipine pretreatment) or at 4 h of recirculation (n=6 with vehicle; n=6 with nifedipine pretreatment; n=6 with nifedipine posttreatment), and the cerebral mitochondrial respiration was measured polarographically. Tissue oxygen tension was evaluated in placental and fetal cerebral tissues (n=5 with vehicle; n=5 with nifedipine pretreatment). The vehicle treated animals showed a significant decrease in mitochondrial activities at the end of ischemia and 4 h of recirculation. Nifedipine attenuates the secondary deterioration at 4 h of recirculation when given just prior to ischemia, but had no neuroprotective activity when given 1 h after the start of recirculation. Nifedipine pretreatment had no influence on oxygen delivery in placenta and fetal cerebrum during and after ischemia. Despite the short therapeutic window, the treatment of nifedipine attenuates the secondary deterioration of cerebral mitochondrial function after transient intrauterine ischemia in fetal rats when given just prior to ischemia. PMID- 12088837 TI - Nuclei and subnuclei gene expression profiling in mammalian brain. AB - Information on the neuroanatomical expression of a given gene is critical to understanding its function in the central nervous system. The integration of laser capture microdissection (LCM), T7-based RNA amplification and cDNA microarrays allows for this information to be simultaneously generated for thousands of genes. To validate this integrative approach, we catalogued the gene expression profiles of seven rat brain nuclei or subnuclei. A hundred cells from the following seven brain nuclei were analyzed: locus coeruleus (LC), dorsal raphe nucleus (DR), parvocellular division (PA) and magnocellular division (MG) of the hypothalamic paraventricular nucleus (PVN) and CA1, CA3 and dentate gyrus (DG) divisions of the hippocampal formation. Of the 2145 genes investigated, 1402 genes (65%) gave a hybridization signal statistically different from the background level that was defined by non-specific hybridizations to 15 different plant genes. Validation of our microarray data on four arbitrarily selected genes was confirmed by Real-Time PCR. Previous research showing expression patterns of 'signature' genes (n=17) for specific brain nuclei are consistent with our findings. For example, as previously shown, enriched mRNA expression encoding the serotonin transporter or tyrosine hydroxylase was found in DR and LC cells, respectively. Interestingly, expression of the serotonin 5-HT(2B) receptor mRNA was also found in DR cells. We confirmed this new finding by in-situ hybridization. The hierarchical clustering analysis of gene expression shows that the two divisions of the PVN (PA and MG) are closely related to each other, as well as the three regions of the hippocampal formation (CA1, CA3 and DG), which also showed similar gene expression profiles. This study demonstrates the importance, feasibility and utility of cellular brain nuclei profiling. PMID- 12088838 TI - Modulation by hyperpolarization-activated cationic currents of voltage responses in human rods. AB - We used the whole-cell patch-clamp recording technique on surgically excised human retina to examine whether human rod photoreceptors express hyperpolarization-activated cationic currents (I(h)) and to analyze the effects of I(h) on rod's voltage responses. Hyperpolarizing voltage steps from a holding potential of -60 mV evoked a slow inward-rectifying current in both rods in retinal slices and isolated rods. The slow inward-rectifying currents induced by hyperpolarization were markedly reduced by 3 mM Cs(+) (a blocker of I(h)) in the bath, but not by 3 mM Ba(2+) (an anomalous rectifier K(+) current blocker) or 1 mM SITS (a Cl(-) current blocker). A concentration-response curve for block by Cs(+) of the inward currents could be fitted by the Hill equation with a half blocking concentration (IC(50)) of 41 microM and a Hill coefficient of 0.91. The time course of the inward current activation was well described at all recorded voltages by the sum of two exponentials. Under current-clamp conditions, injection of steps of current, either hyperpolarizing or depolarizing, elicited an initial rapid voltage change that was followed by a gradual decay in the voltage response. The decay in the voltage responses was eliminated by bath application of 3 mM Cs(+). The voltage dependence, pharmacology, and kinetics of the slow inward-rectifying currents described above suggest that human rods express I(h). We suggest that I(h) becomes activated in the course of large hyperpolarizations generated by bright-light illumination and may modify the waveform of the photovoltage in human rods. PMID- 12088839 TI - Microtubule-associated protein 1B is involved in the initial stages of axonogenesis in peripheral nervous system cultured neurons. AB - Neuronal process extension is dependent on the reorganisation of the cytoskeleton, in particular microtubules and microfilaments, and one of the ways in which microtubules are regulated is by a group of microtubule-associated proteins called MAPs. MAP1B, the first MAP to be expressed in developing neurons, has been shown to play an important role during axonogenesis. Previously, we have shown that a phosphorylated isoform of MAP1B is involved in maintaining growth cone microtubules in a dynamically unstable state. In order to further investigate the role of MAP1B during axonogenesis we have cultured dorsal root ganglion (DRG) neurons from a MAP1B deficient mutant mouse. These mice express only trace amounts of MAP1B, have defects in the development of their nervous system and die perinatally. Cultured DRG neurons from MAP1B deficient mice show a reduction in axon elongation and an increase in growth cone area. The reduction in axon elongation is most likely to occur due to an inhibition in the early stages of axonogenesis. Using time-lapse video we have verified that during the first 2 h after plating, MAP1B deficient neurones extend their axons with an average speed that is half the speed of control neurones. These results support the participation of MAP1B during the initial stages of axonogenesis. PMID- 12088840 TI - Quantitative autoradiography of adenosine receptors and NBTI-sensitive adenosine transporters in the brains and spinal cords of mice deficient in the mu-opioid receptor gene. AB - There is a large body of evidence indicating important interactions between the adenosine and opioid systems in regulating pain at both the spinal and supraspinal level. Mice lacking the mu-opioid receptor (MOR) gene have been successfully developed and the animals show complete loss of analgesic responses to morphine as well as differences in pain sensitivity. To investigate if there are any compensatory alterations in adenosine systems in mutant animals, we have carried out quantitative autoradiographic mapping of A(1) and A(2A) adenosine receptors and nitrobenzylthioinosine (NBTI) sensitive adenosine transporters in the brains and spinal cords of wild type, heterozygous and homozygous mu-opioid receptor knockout mice. Adjacent coronal sections were cut from the brains and spinal cords of +/+, +/- and -/- mice for the determination of binding of [3H]DPCPX, [3H]CGS21680 or [3H]NBTI to A(1) and A(2A) adenosine receptors and NBTI-sensitive adenosine transporters, respectively. A small but significant reduction in [3H]DPCPX and [3H]NBTI binding was detected in mutant mice brains but not in spinal cords. No significant change in A(2A) binding was detected in mu-opioid receptor knockout brains. The results suggest there may be functional interactions between mu-receptors and A(1) adenosine receptors as well as NBTI sensitive adenosine transporters in the brain but not in the spinal cord. PMID- 12088841 TI - Lesion induced new neuron incorporation in the adult hypothalamus of the avian brain. AB - Cell loss in most adult vertebrate brain regions is thought to be irreversible. Here, we explore the effects of electrolytic lesions on the induction of cell proliferation and newborn neurons in the ventromedial nuclei (VMN) of the hypothalamus in young and adult ring doves. The hypothalamus does not normally recruit new neurons. Bromodeoxyuridine (BrdU) and tritiated thymidine ([3H]Thy) were used to identify cells born before and after bilateral electrolytic lesions. Hu and NeuN were used to identify neurons. TUNEL test for apoptosis and 3A7 antibodies were used to identify morphological changes of pre-existing cells. Lesions produced significantly more newborn cells in the subventricular zone (SVZ). The rate of cell proliferation peaked at 7-14 days postlesion. A fraction of these newborn cells were neuronal precursor and began to migrate away along the radial glial fibers 2 weeks after lesion. During this period, the outer area of the lesion site was marked with massive apoptosis and re-expression of radial glial-like fibers. In birds that survived 5 months, we found newly differentiated neurons in the outer area of the lesion site. We conclude that electrolytic lesion can invoke neuronal recruitment in the adult hypothalamus. We further suggest that lesion-induced apoptosis and re-expression of developmental mechanisms might be involved in the recruitment process. PMID- 12088843 TI - Combined systemic administration of morphine and magnesium sulfate attenuates pain-related behavior in mononeuropathic rats. AB - The response to opioids is reduced in neuropathic pain states. We examined the effect of the combination of morphine (0.1 mg/kg) and magnesium sulfate (125 mg/kg) on behavioral signs of neuropathic pain in spinal nerve ligated rats. Administered alone, neither drug produced any effect, but the combination exerted a significant anti-allodynic effect, which was partially reversed by naloxone. These results suggest that combining low doses of magnesium sulfate with opiates might be an alternative in treating neuropathic pain, with reduced risk of side effects. PMID- 12088842 TI - Superior colliculus firing changes after lesion or electrical stimulation of the subthalamic nucleus in the rat. AB - Recent data have suggested a critical role for the basal ganglia in the remote control of epileptic seizures. In particular, it has been shown that inhibition of either substantia nigra pars reticulata or subthalamic nucleus as well as activation of the superior colliculus suppresses generalized seizures in several animal models. It was previously shown that high frequency stimulation of the subthalamic nucleus, thought to act as functional inhibition, stopped ongoing non convulsive generalized seizures in rats. In order to determine whether high frequency stimulation of the subthalamic nucleus involved an activation of superior colliculus neurons, we examined the effects of subthalamic nucleus manipulation, by either high frequency stimulation or chemical lesion, on the spontaneous electrical activity of superior colliculus neurons. Acute high frequency stimulation of the subthalamic nucleus (frequency 130 Hz) induced an immediate increase of unitary activity in 70% of responding cells, mainly located within the deep layers, whereas a reduction was observed in the remaining 30%. The latter responses are dependent on the intensity and frequency of the stimulation. Unilateral excitotoxic lesion of the subthalamic nucleus induced a delayed and transient decrease of superior colliculus activity. Our data suggest that high frequency stimulation of the subthalamic nucleus suppresses generalised epileptic seizures through superior colliculus activation. PMID- 12088845 TI - Demonstration of expression of mRNA for particular AMPA and kainate receptor subunits in immature and mature cultured rat calvarial osteoblasts. AB - Reverse transcription polymerase chain reaction revealed expression of mRNA for particular subunits of ionotropic glutamate receptors (iGluR) in primary cultures of rat calvarial osteoblastic cells under immature to mature states. These included GluR3, KA1 and KA2 subunits, in addition to NR1 and NR2D subunits. These results suggest that glutamate may play an unidentified role in mechanisms associated with cellular development through particular subunits of iGluR in rat calvarial osteoblasts. PMID- 12088844 TI - Acute and long-term effects of the 5-HT2 receptor antagonist ritanserin on EEG power spectra, motor activity, and sleep: changes at the light-dark phase shift. AB - Parallel effects of a single injection of the 5-HT(2) receptor antagonist ritanserin on EEG power spectra, sleep and motor activity were measured for a 20 h period in freely moving Sprague-Dawley rats. Ritanserin (0.3 mg/kg, i.p.), administered at light onset (passive phase), caused an immediate transient increase in the EEG power density in the low frequency range (0.25-6 Hz, mainly delta activity) and a depression in the high frequency range (27-30 Hz) accompanied by a decrease in vigilance and light slow wave sleep (SWS-1), intermediate stage of sleep and increase in deep slow wave sleep (SWS-2) compared to control treatment. All these effects were over 8 h after the injection. Twelve hours after the injection, at dark onset (active phase), there was a marked increase in vigilance and motor activity and decrease in SWS-1 and spindle frequency activity in the control animals, but all these changes were diminished by ritanserin treatment. These effects resulted in a significant relative increase in the intermediate band (peak: 12-15 Hz) of the EEG power spectra and thus, a relative increase in thalamo-cortical synchronization caused by ritanserin at dark onset. Because ritanserin is a selective 5-HT(2) receptor antagonist, we conclude that under physiological conditions serotonin increases EEG desynchronization and produces an increase in vigilance level and motor activity by tonic activation of 5-HT(2) receptors. This regulatory mechanism plays an important role in the waking process, and the appearances of its effects in the light and dark phase are markedly different. PMID- 12088846 TI - Vasopressin-induced cytoplasmic and nuclear calcium signaling in cultured cortical astrocytes. AB - We sought to determine whether vasopressin V(1a) receptor (V(1a)R) mRNA detected in cortical astrocytes [Brain Res. Mol. Brain Res. 45 (1997) 138] was translated into functional receptors by investigating the effector calcium signaling cascade associated with the vasopressin V(1a) receptor subtype. Analysis of intracellular calcium dynamics using the calcium-sensitive dye fura-2 AM indicated that exposure of cortical astrocytes to V(1) vasopressin receptor agonist, [Phe(2),Orn(8)]-oxytocin, induced a marked dose-dependent increase in intracellular calcium which was abolished by depletion of extracellular calcium. V(1) agonist treatment induced a rapid increase in calcium signal in both the cytoplasm and nucleus, which was followed by an accumulation of the calcium signal in the nucleus, suggesting translocation of cytoplasmic calcium into the nucleus. The nuclear calcium signal was sustained for several seconds followed by translocation back to the cytoplasm. Following the nuclear-to-cytoplasmic calcium translocation, total free intracellular calcium concentration decreased. The dynamic calcium cytoplasmic and nuclear localization was confirmed by laser scanning confocal microscopy coupled with the calcium-sensitive dye fluo-3 AM. To determine the source of calcium, V(1) agonist-induced (45)Ca(2+) uptake and [(3)H]IP(1) accumulation were investigated. V(1) agonist induced significant and rapid uptake of (45)Ca(2+) and a significant dose-dependent increase in [(3)H]IP(1) accumulation in cortical astrocytes. To our knowledge, this is the first documentation of a vasopressin receptor-induced calcium signaling cascade in cortical astrocytes and the first documentation of vasopressin receptor induction of nuclear calcium signaling. PMID- 12088847 TI - Selective lesion of cholinergic neurons in the medial septum by 192 IgG-saporin impairs learning in a delayed matching to position T-maze paradigm. AB - This study examined whether selective destruction of cholinergic neurons in the medial septum impairs acquisition of a delayed matching-to-position (DMP) spatial memory task. Either the selective immunotoxin 192 IgG-saporin (SAP; 0.22 or 1.0 microg) or the non-selective excitatory neurotoxin ibotenate (IBO; 5 microg), was infused directly into the medial septum of rats. Both doses of SAP, but not IBO, significantly impaired acquisition of the DMP task and blunted the initial alternating behavior of the rats in the T-maze. Histochemical staining revealed that both doses of SAP produced a near complete depletion of choline acetyltransferase (ChAT)-positive neurons in the medial septum. Some loss of parvalbumin staining was observed following administration of the higher dose, but not the lower dose, of SAP. In contrast, IBO produced a nearly complete depletion of parvalbumin-positive staining throughout the medial septum. IBO also produced a loss of ChAT-positive neurons and considerable local damage in the medial septum around the area of injection; however, many ChAT-positive neurons in the medial septum distal to the injection remained. A significant correlation between the number of days to reach criterion and ChAT activity in the frontal cortex and hippocampus was observed. The results suggest that low doses of SAP can be used to selectively destroy cholinergic neurons in the medial septum, and that selective destruction of these neurons significantly impairs acquisition of the DMP task. We propose that acquisition of the DMP task is a sensitive behavioral assay for the selective loss of basal forebrain cholinergic projections. PMID- 12088848 TI - Human urocortin II: mild locomotor suppressive and delayed anxiolytic-like effects of a novel corticotropin-releasing factor related peptide. AB - Recently, human urocortin II (hUcn II), a member of the corticotropin-releasing factor (CRF) peptide family, was identified. The following experiments sought to compare the effects of this novel CRF-related peptide versus those of ovine CRF (oCRF) on locomotor activation and anxiety-related behavior, using the locomotor activity test and the elevated plus maze, respectively. To examine locomotor activity during the active (dark) and inactive (light) phases, rats were intracerebroventricularly (i.c.v.) injected with 0, 0.1, 1.0 or 10 microg of hUcn II (n=8/group active; n=6-9/group inactive) or oCRF (n=8/group active; n=8/group inactive) 2 h after the onset of their respective testing phase and monitored for 3 (inactive) or 5 (active) h. To compare the effects of CRF-related peptides on exploration of the elevated plus maze, rats were pretreated (i.c.v. 0, 0.1, 1.0 or 10 microg) with hUcn II (n=7-11/group) or oCRF (n=7-10/group), 10 min prior to testing. Delayed effects in the elevated plus maze were examined in rats injected with 1.0 microg of hUcn II (n=8/group) or oCRF (n=6-8/group), or vehicle (n=8/group) 1, 4 or 6 h before testing. In contrast to the activational effects of oCRF, hUcn II mildly suppressed locomotor activity during the inactive phase. hUcn II did not acutely affect open arm exploration in the elevated plus maze, whereas oCRF decreased this measure. However, hUcn II increased open arm exploration 4 h after injection. Thus, hUcn II exhibits mild motor suppressive effects and delayed anxiolytic-like effects, suggesting a time-dependent role for hUcn II in the regulation of stress-related behavior. PMID- 12088849 TI - Estrogen and opioids interact to modulate the locomotor response to cocaine in the female rat. AB - Estrogen is known to modulate the behavioral response to cocaine; however the mechanisms by which this is accomplished is unknown. In this study we examine one possible candidate, the endogenous opioid system. Adult Sprague-Dawley rats were ovariectomized (OVX), half received Silastic implants with estradiol benzoate (OVX-EB), the other half received empty implants (OVX). After 1 week, spontaneous locomotor and stereotyped activity was measured for 60 min using an automated system. On day 2, locomotor activity was recorded for 30 min. Rats were injected with saline (SAL) or naloxone (NAL) (2 mg/kg, i.p.) and activity measured for the next 20 min. Each of these groups were further subdivided, one that received a saline injection (SAL) and another that received a cocaine injection (COC) (15 mg/kg, i.p.). Locomotor and stereotyped activities were recorded for 60 min. This resulted in the following injection groups: SAL-SAL, NAL-SAL, SAL-COC and NAL COC. During habituation, OVX rats displayed an overall higher level of activity than OVX-EB rats. Similar to what is observed in males, naloxone significantly reduced locomotion and stereotyped behavior but only in OVX rats. Estrogen administration to OVX rats abolished the effect of naloxone. Surprisingly, when naloxone was administered prior to cocaine, an increase in cocaine-induced locomotor and stereotyped activity was observed, but only in OVX-EB rats. These results indicate that opioid modulation of cocaine-induced locomotor and stereotype activity in the female differs from that reported in the male. In addition in the female, the effect of opioids on cocaine-induced locomotor behavior is dependent on plasma levels of estrogen. PMID- 12088850 TI - Sero-epidemiological assessment and diagnosis of visceral leishmaniasis in an endemic locality using Fast Agglutination Screening Test (FAST). AB - The Fast Agglutination Screening Test (FAST) was employed on sera obtained from an endemic area of visceral leishmaniasis in southwestern Ethiopia, in February 2000. The study involved (i) active case detection among 1575 residents of two villages; and (ii) passive case detection in an outpatient clinic. Sera of 1587 individuals, including 143 sera of previously treated VL patients, were tested. Based on the size of agglutination mat, the FAST results were read qualitatively as non-reactive (-), weakly reactive (1+), moderately reactive (2+) and highly reactive (3+). All FAST reactive sera were re-tested with the Direct Agglutination Test (DAT). After clinical screening of 1625 individuals, 61 individuals with signs and symptoms of early or late VL were found; 26 sera were FAST positive. Twenty-two of these suspected VL cases were subjected to parasitological examination using lymph node aspirates. Eighteen (81.8%) were confirmed either by demonstration of amastigotes in smears or promastigotes in NNN cultures. FAST reactive anti-leishmanial antibodies were detected in 4.5% of untreated and 70.6% of previously treated patients. Forty-five sera of 1390 previously untreated asymptomatic individuals (3.2%) were found to be FAST positive. This report demonstrates that FAST is a rapid and cost-effective screening test for the diagnosis and sero-epidemiological surveillance of visceral leishmaniasis. PMID- 12088851 TI - Effect of Trypanosoma cruzi released antigens binding to non-infected cells on anti-parasite antibody recognition and expression of extracellular matrix components. AB - It has been proposed that antigens released by Trypanosoma cruzi sensitize vertebrate cells leading to their destruction by the immune response raised against the parasite. Here, we characterized antigens released by trypomastigotes of T. cruzi that bind to non-infected cells and investigated biological consequences of this adsorption. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of antigens released by [(35)S]-methionine labeled parasites revealed the presence of polypeptides mainly ranging from 85 to 170 kDa that were specifically recognized by sera from chronically T. cruzi infected rabbits. Polypeptides of 85-110 and 160-170 kDa bound to non-infected epithelial, fibroblast and muscle mammalian cell lines, which thus became targets for anti-T. cruzi antibody binding. Cysteine-proteinase, but not trans-sialidase, was detected among the cell-bound antigens, and purified cysteine-proteinase was adsorbed to non-infected cells. Immunoelectron microscopic studies showed that parasite antigens were mainly released as membrane vesicles that adhered to membrane microvilli and were internalized by mammalian cells. We provide evidence that adsorption of parasite antigens induced an increase in expression of extracellular matrix (ECM) components (fibronectin, laminin and type I collagen) by sensitized cells. Thus, our data reinforce the idea that in vivo T. cruzi released antigens might be involved in the establishment of inflammation, sensitizing non-infected host cells and triggering an immune response against parasite antigens. Further, our data showed that antigen sensitization modulates biological cell functions as ECM expression that could mediate cell-cell or parasite-host cell interactions, contributing to the establishment of inflammation. PMID- 12088852 TI - Sensitivity of Plasmodium vivax to chloroquine in Sa Kaeo Province, Thailand. AB - Following a recent, abrupt local increase in the incidence of vivax malaria, a study was conducted in order to evaluate the efficacy of chloroquine for the treatment of 26 adult patients with acute vivax malaria in Sa Kaeo Province, Thailand. The chloroquine sensitivity of Plasmodium vivax has been assessed in parallel, using a growth inhibition method. Blood samples for the in vitro tests were taken prior to the administration of the standard treatment with chloroquine -in total 25 mg base/kg over 3 days--and primaquine 0.25 mg base/kg once daily for 14 days. The efficacy has been assessed according to the WHO standard in vivo test. The cure rate was 100%. No recrudescence was observed during the follow-up period of 28 days. The mean fever clearance time (FCT) was 40 h, the mean parasite clearance time (PCT) was 49 h. Mean IC(50) and IC(90) of the parasites were 28 and 171 nM, respectively. These results show that local P. vivax is still sensitive to chloroquine. The epidemic outbreak was therefore obviously not due to the presence of chloroquine-resistant P. vivax. PMID- 12088854 TI - Elevated levels of nitric oxide and low levels of haptoglobin are associated with severe malarial anaemia in African children. AB - Severe malarial anaemia (SA) is a major complication of malaria and an important cause of child mortality and morbidity. However, the pathogenesis behind SA is poorly understood. Nitric oxide (NO) is known to play a protective role against clinical malaria but is also suggested to have a pathogenic role in cerebral malaria (CM). Erythrophagocytosis by splenic macrophages has been implicated in the pathogenesis of SA. In this study, plasma levels of NO, neopterin, haptoglobin and C-reactive protein (CRP) were measured in paediatric patients with CM, n=77, SA (n=28) and uncomplicated malaria (UM n=53). Haptoglobin levels were significantly lower in SA (median (interquartile range) 25 (17-59) mg/l) than in both CM and UM (40 (24-80) mg/l and 110 (60-160) mg/l, respectively, P<0.001). In contrast, NO levels were higher in SA (38 (28-51) micromol/l) than in CM and UM (21 (15-32) micromol/l and 10.3 (5.6-17) micromol/l, respectively, P<0.001). A significant negative correlation between haptoglobin and NO was seen in the SA group. No such correlation was observed within the UM or CM groups. No significant differences in neopterin levels were observed between any of the three groups, neither was there any correlation between parasitaemias and neopterin levels. The low haptoglobin and high levels of NO in this SA group may contribute to haemolysis. Taken together our results support the hypothesis that immune-mediated erythrocyte destruction is involved in the pathogenesis of malarial anaemia. PMID- 12088853 TI - Infection with Schistosoma mansoni correlates with altered immune responses to Ascaris lumbricoides and hookworm. AB - Studies were performed on humoral and cellular immune responses of patients from areas in Brazil endemic for hookworm and Ascaris lumbricoides, and either endemic or non-endemic for Schistosoma mansoni. Humoral and cellular responses were evaluated by enzyme-linked immunosorbant assay (ELISA) and peripheral blood mononuclear cell (PBMC) proliferation assays against larval hookworm antigens, A. lumbricoides egg antigens, and soluble egg antigens (SEA) or soluble whole adult antigenic preparation (SWAP) from S. mansoni. Patients from S. mansoni-endemic areas, who currently had only hookworm or Ascaris infections, expressed lower humoral and cellular responses to hookworm or Ascaris antigens, respectively, than did their counterparts from areas not endemic for S. mansoni. Individuals from S. mansoni endemic area, although without detectable S. mansoni infection, do mount humoral and cellular responses to SEA and SWAP. This group of individuals has been probably in contact with S. mansoni antigens, since the groups harboring A. lumbricoides or hookworm infections from non-S. mansoni endemic areas do not have detectable anti-S. mansoni responses. PBMC proliferative responses discriminated well between patients with active hookworm infections versus ascariasis, if they were from areas not endemic for S. mansoni. PMID- 12088855 TI - Spatial and intensity-dependent variations in associations between multiple species helminth infections. AB - Estimated associations between infections with different helminth species can be used to predict the proportion of a population infected with multiple species infections. This is an important measure of disease burden, as those with multiple infections are often at an increased risk of morbidity. In this paper, we investigate variation amongst the estimated associations between Ascaris lumbricoides, Trichuris trichiura and hookworm, over a number of different spatial levels among schoolchildren in Cameroon. Associations between species were largely homogeneous within districts, provinces and ecological zones, although variation between these regions was identified, implying that a single measure of association may not be appropriate in different epidemiological settings. Further data collected amongst school children in Kenya and Uganda were analysed, to assess the dependence of the associations on the intensity of infection. It was found that the strength of the association between A. lumbricoides and T. trichiura increased with intensity, such that those with more intense infections with one species are increasingly likely to harbour concurrent intense infections with the other species. The implications of these results are discussed in relation to the estimation of the disease burden due to multiple helminth species. PMID- 12088856 TI - Morbidity and nutritional impairment in relation to CD4 count in a Zambian population with high HIV prevalence. AB - The HIV epidemic has greatly increased morbidity in many African cities and severe undernutrition is a prominent feature of the clinical presentation. However, there is little information about the relationship of morbidity or nutritional status to immune damage at a population level. We report a cross sectional study of morbidity and nutritional status in relation to CD4 count in an impoverished urban community in Lusaka, Zambia, at enrollment into a longitudinal study. Over a 2 month period in 1999, 261 (52%) of 506 adults resident in one area were interviewed and examined. Of 186 adults who consented to testing, 33 (51%) of 65 who were HIV seropositive reported symptoms of disease compared to 39 (32%) of 121 who were HIV seronegative (OR 2.2, 95%CI 1.1-4.2; P=0.02). Peripheral blood CD4 counts in HIV seronegative individuals were broadly similar to norms in developed countries, but 8 (7%) had CD4 counts below 500 cells/microl. Morbidity in HIV seropositive adults was dominated by tuberculosis (n=11), other respiratory infections (5) or persistent diarrhoea (4), and affected individuals had a wide range of CD4 counts. Nutritional impairment was evident in HIV seropositive adults with clinical evidence of opportunistic infection (OI), not those with asymptomatic HIV infection. Unexpectedly, we also noted that systolic blood pressure was reduced progressively in HIV infection and in those with OI. In conclusion, HIV-related morbidity was dominated by a small number of treatable infectious diseases occurring over a wide range of CD4 count. Nutritional impairment was associated with OI. PMID- 12088857 TI - Evaluation of an antigen from Taenia crassiceps cysticercus for the serodiagnosis of neurocysticercosis. AB - We report here the evaluation of an antigen from Taenia crassiceps cysticercus as a potential reagent in an enzyme-immunoelectrotransfer blotting assay (EITB) and an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of neurocysticercosis (NC) using clinical specimens obtained from patients in different phases of the disease. Serum and cerebrospinal fluid (CSF) samples from 64 patients suspected of having NC according to clinical manifestation and brain computed tomography were tested by ELISA with Taenia solium total saline antigen (ELISA-Tso) and by immunoblotting with T. crassiceps glycoproteins antigen (EITB gpTcra). Forty-five serum samples were also tested immunoblotting with T. solium glycoproteins antigen (EITB-gpTso) and 30 were tested by ELISA with T. crassiceps 14 kDa glycoprotein (ELISA-gp14Tcra). Serum samples from apparently healthy individuals without any parasitic disease and from patients with other parasitic diseases were included as controls. The results of ELISA-Tso analysis with CSF obtained from 64 patients with NC showed that 53 (83%) were reactive. EITB-gpTcra analysis with serum from the same group of patients showed a sensitivity of 91%. Results of EITB-gpTso and EITB-gpTcra analysis with serum samples demonstrated an agreement of 100% between both tests. ELISA-gp14Tcra was positive in 23 (77%) sera, 22 with paired CSF positive. When ELISA-gp14Tcra results were compared to EITB-Tso results, a relative sensitivity of 95% was observed. All serum samples from the control group were negative in ELISA-gp14Tcra and only one serum from an individual with Taenia saginata was reactive in this assay, showing a specificity of 99% for ELISA-gp14Tcra. This fraction was purified in only one step with a good yield for use in immunoassays. We suggest that the gp14Tcra antigen can be used for detecting anti-cysticercus antibodies in serum samples for epidemiological investigation purposes and also for diagnostic screening of NC patients. PMID- 12088858 TI - Molecular characterization of Vibrio cholerae O1 outbreak strains in Miri, Sarawak (Malaysia). AB - Bacterial resistance to various antimicrobial agents is common in area with high usage of antibiotics. In this study, the data on antimicrobial susceptibility patterns of Vibrio cholerae O1 from patients during an outbreak period was found to be high but variable rates of multidrug resistance. Thirty-two of 33 V. cholerae isolates harboured the tcp, ctx, zot and ace genes, suggesting their possible roles in the outbreak cases. We analyzed the molecular diversity of a total of 33 strains of V. cholerae O1 isolated from 33 patients between November 1997 and April 1998 using random amplified polymorphic DNA (RAPD) analysis. The 30 typable isolates could be separated into four major clusters containing 5, 17, 2 and 6 isolates, respectively. However, no particular RAPD pattern was predictive of a particular pattern of antibiotic susceptibility. The findings of this study showed that multiple clones seemed to be responsible for cases in the outbreaks in the study area. PMID- 12088859 TI - Albendazole-soybean oil emulsion for the treatment of human cystic echinococcosis: evaluation of bioavailability and bioequivalence. AB - A single 12.5 mg/kg dose of albendazole (Abz) in tablet form (AbzT) followed 2 weeks later by an equivalent dose of Abz emulsified in 30% soybean oil (AbzE) was administered orally 2 h after the first morning meal to 7 male adult patients with cystic echinococcosis caused by Echinococcus granulosus. Serum samples were taken 1, 3, 5, 7, 8, 9, 11, 14, 18, 24, 36, and 48 h post medication from each patient to measure the serum concentrations of albendazole sulfoxide (AbzSOX), the principal bioactive metabolite of Abz. AbzSOX concentrations were measured by reverse phase HPLC. The data were subjected to pharmacokinetic analysis to compare the relative bioavailability and bioequivalence of AbzT and AbzE. The results demonstrated that the mean peak concentrations (C(max)) for AbzT and AbzE were 1.06+/-0.38 mg/l and 1.71+/-0.47 mg/l, respectively; the area under the concentration-time curves (AUC) were 13.24+/-4.93 mg x h/l and 21.01+/-7.54 mg x h/l, respectively. The relative bioavailability of AbzE was F(Flu)=1.59. Two one sided tests procedure and (1-2 alpha) 90% confidence interval methods were used to evaluate the bioequivalence of AbzE and AbzT. The results demonstrated that the bioavailability of AbzE was greater than AbzT. PMID- 12088860 TI - A novel and sensitive method to monitor helminth infections by faecal sampling. AB - The Kato-Katz technique is the method routinely used for diagnosing human schistosomiasis mansoni by estimating faecal egg burdens. To improve the sensitivity of faecal diagnosis, we established and validated a novel separation technique based upon the greater density of viable schistosome eggs relative to faecal material. Subsequently, it was used for faecal examination of 27 schistosomiasis patients in El-Sharkia, Egypt, with Kato-Katz smears as criterion standard. Low intensity infections (<100 eggs/g) were only detected by our technique. Moreover, triple Kato-Katz analysis on consecutive samples still missed 7.4% of all human patients, whereas the new method diagnosed 100% of samples correctly on second analysis. We conclude that in endemic areas many patients are being systematically missed by routine diagnosis. Moreover, the sensitivity of our method allows its use in proposed pre-clinical and clinical vaccine trials in non-human primates and humans, where reliable estimates of faecal egg counts are essential. PMID- 12088861 TI - 17beta-Estradiol upregulates distinct maxi-K channel transcripts in mouse uterus. AB - The mouse maxi-K channel transcript undergoes alternative splicing to produce isoforms differing in sensitivity to intracellular regulators. We hypothesized that 17beta-estradiol could induce myometrial maxi-K channel transcripts to differentially splice. Polymerase chain reaction demonstrated two products at site D in mice injected with either 8.5 microg of 17beta-estradiol for 4 days or a vehicle control. Splicing of site D is known to modulate the sensitivity of the maxi-K channel to calcium and voltage. RNase protection analyses revealed that the alpha subunit transcript, and an exon encoding 59 amino acids at site D that enhances Ca(2+)- and voltage-sensitivity, are upregulated approximately 1.4-fold after 17beta-estradiol stimulation however, the insertless isoform of this transcript is enhanced approximately 5-fold. Immunoblotting demonstrates that the total maxi-K channel alpha subunit expression mimics transcript regulation. These findings verify that maxi-K channel transcripts are differentially spliced by 17beta-estradiol, which may contribute to stoichiometric changes in isoform expression during pregnancy. PMID- 12088862 TI - Mechanism of angiotensin II-induced phospholipase D activation in bovine adrenal glomerulosa cells. AB - Based on previous data demonstrating activation of phospholipase D (PLD) in response to angiotensin II (AngII), we have hypothesized a role for PLD in mediating aldosterone secretion from bovine adrenal glomerulosa cells. In this study we demonstrate that a PLD-generated signal(s) is required for the AngII elicited secretory response, since interfering with lipid second messenger formation using a primary alcohol inhibited AngII-induced aldosterone secretion, but not that elicited by incubation with a hydrophilic cholesterol analog, 22(R) hydroxycholesterol, which bypasses signaling pathways. Three mechanisms for hormonal activation of PLD have been described in other systems: direct receptor coupling, activation through protein kinase C (PKC) and a combination of these two mechanisms. Our results indicate that the PKC activator, phorbol 12-myristic 13-acetate (PMA), is able to activate PLD, and that receptor engagement is apparently not necessary for PLD activation in response to this agent. Maximal doses of AngII and PMA produced no additive effect on PLD activation, suggesting that these two agents function through a common PKC pathway. This interpretation was confirmed by the ability of a PKC inhibitor, Go 6976, to inhibit partially AngII-induced PLD activation. Finally, treatment with the calcium ionophores A23187 or ionomycin or the calcium channel agonist BAY K8644 had no effect on PLD activity. Likewise, inhibiting calcium influx with high-dose nitrendipine affected neither basal PLD activity nor that stimulated by AngII. Thus, our results suggest a role for PKC, independent of calcium influx, in mediating AngII induced PLD activation in glomerulosa cells. PMID- 12088863 TI - Three mRNA species for mammalian-type gonadotropin-releasing hormone in the brain of the eel Anguilla japonica. AB - A variant mRNA species for mammalian-type gonadotropin-releasing hormone (mGnRH) that retains first intron has recently been found in the brain of a primitive teleost, the eel Anguilla japonica. Here we have found that this mRNA species is generated from an alternative upstream transcription start site of the mGnRH gene. In addition, the upstream start site produces another mRNA species that lacks first intron. Both of these two variant mRNA species, however, have the same open reading frame as the downstream mGnRH mRNA, whose introns are all spliced out, and therefore, all three mGnRH mRNA species expressed in the eel brain would translate a common prepro-mGnRH polypeptide. Quantitative analysis revealed that the upstream mRNAs are less abundant accounting for not more than 1% of the total mGnRH mRNAs. When testosterone was administered (1 mg per fish per day) continuously for 5 days to juvenile male eels, both upstream and downstream mGnRH mRNA species in the brain were up-regulated. However, the level of the upstream mRNAs increased by 3-5-fold after 12 days of the treatment, whereas that of the downstream mRNA showed a more modest increase (about 2-fold) although this increase was more rapid. These results demonstrate that both upstream and downstream mGnRH mRNAs in the eel brain are under the hormonal control, but they are regulated in different and independent manners. PMID- 12088865 TI - Insulin but not insulin-like growth factor-1 promotes the primordial to primary follicle transition. AB - A critical step in ovarian biology is the transition of the developmentally arrested primordial follicle to the growing primary follicle. The current study utilizes a rat ovarian organ culture system to investigate the role of insulin and insulin-like growth factor-1 (IGF-1) in this process. Four-day-old rat ovaries were cultured and the degree of primordial to primary follicle transition measured. Insulin increased the primordial to primary follicle transition 30% over control with a half maximal effective concentration (EC50) between 2.5 and 5 ng/ml. IGF-1 did not cause an increase in the primordial to primary follicle transition at concentrations up to 100 ng/ml. Ovaries were also treated with epidermal growth factor (EGF) and hepatocyte growth factor (HGF) and neither had an effect on the primordial to primary follicle transition. Ovaries were treated with insulin in conjunction with other factors known to promote the primordial to primary follicle transition in order to discern any potential synergistic effects. Previous experiments have shown that kit ligand (KL), basic fibroblast growth factor (bFGF) and leukemia inhibitory factor (LIF) promote the primordial to primary follicle transition. Insulin was shown to have an additive effect with KL and LIF, but not bFGF. The fact that insulin can influence the primordial to primary follicle transition at low concentrations (i.e. 5 ng/ml) and that IGF-1 has no effect suggests that insulin is acting at the insulin receptor, not the IGF-1 receptor. The observation that insulin has an additive effect with KL and LIF, but not bFGF, suggests the insulin's site of action is likely the oocyte. In summary, observations suggest that insulin acts as an endocrine type factor to help coordinate primordial to primary follicle transition at the level of the oocyte. The significance of the observations in relation to diabetes and female infertility is discussed. PMID- 12088864 TI - Mechanisms of bradykinin-induced glucagon release in clonal alpha-cells In-R1-G9: involvement of Ca(2+)-dependent and -independent pathways. AB - The mechanisms by which bradykinin (BK) increases glucagon release were investigated. BK (0.1-10 microM) increased [Ca(2+)](i) and glucagon release in clonal alpha-cells In-R1-G9. BK-induced glucagon release was lower in the absence than in the presence of extracellular Ca(2+), but it still increased glucagon release while [Ca(2+)](i) was stringently deprived. Depletion of intracellular Ca(2+) store with thapsigargin abolished both the BK-induced Ca(2+) peak and sustained plateau. Microinjection of heparin abolished BK-induced Ca(2+) release. Pertussis toxin (PTX) did not block BK-induced [Ca(2+)](i) increase or glucagon release. U-73122 (8 microM), a phospholipase C (PLC) inhibitor, abolished BK induced increases in [Ca(2+)](i), but only reduced BK-induced glucagon release by 40%. A phospholipase D (PLD) inhibitor zLYCK reduced BK-induced glucagon release by 60%. The combination of U-73122 and zLYCK abolished BK-induced glucagon release. Both SK&F 96365, a receptor-operated Ca(2+) channel (ROC) blocker and nimodipine, an L-type Ca(2+) channel blocker, reduced BK-induced [Ca(2+)](i) increase and glucagon release. These findings suggest that BK increase glucagon release through a PTX-insensitive G protein and both Ca(2+)-dependent and independent pathways. The Ca(2+)-dependent pathway is attributable to PLC activation. PLC catalyzes IP(3) formation, inducing Ca(2+) release from the endoplasmic reticulum, which, in turn, triggers Ca(2+) influx via both ROCs and L type channels. PLD activation may be involved in Ca(2+)-dependent and/or independent pathway. PMID- 12088866 TI - Progesterone receptor activates its promoter activity in human endometrial stromal cells. AB - Previous studies have shown that progestin increases the content of progesterone receptor (hPRA and hPRB) and the hPR mRNA during decidualization of human endometrial stromal cells suggesting that endogenous hPR enhances the transcription of the hPR gene. In the present study, we provide evidence that hPR regulates the promoter activity mediated through an active Sp1 site. In stromal cells treated with medroxyprogesterone acetate, the promoter activity was significantly increased when cells were co-transfected with hPR expression vector. Progressive deletion analysis showed that the highest activity was derived from the promoter region between -55 and +31 bp. Transactivation by hPR was dose dependent. The capacity of hPRA was stronger than that of hPRB. The ligand binding domain, but not DNA binding domain of the hPR was required for the transactivation. The proximal promoter region lacks a canonical progesterone response element. Instead, an active Sp1 site (-49 to -43 bp) has been confirmed. Mutation of the Sp1 site eliminated the effect of hPR activation. The promoter activity was increased by over expression of Sp1, whereas Sp3 had no effect. Electrophoretic mobility shift assay showed that the promoter region between -55 and +31 bp bound to Sp1 family proteins, Sp1 (C2 complex) and Sp3 (C1 and C3 complexes) identified by antibodies to Sp1 and Sp3. Sp1 complex formed by extracts of stromal cells was less intense than that formed by progestin decidualized stromal cells. Sp1/DNA binding was enhanced when stromal cell extracts were incubated with calf intestine alkaline phosphatase (CIP) suggesting that dephosphorylation of Sp1 enhances the DNA binding. Addition of protein kinase inhibitor, H-89 or H-7, enhanced the hPR stimulated promoter activity. Western blot analysis showed that endometrial stromal/decidual cell extracts contained a wide band of Sp1 spanning from approximately 105 to 96 kDa and was resolved into one band at 96 kDa by CIP. Decidual cell extracts are abundant with the 96 kDa Sp1. In addition, the 96 kDa Sp1 was co-precipitated with ligand activated hPRA or hPRB in the decidual cell nuclear extracts. These data suggest that dephosphorylated Sp1, abundant in decidual cells, enhances the binding to both DNA and hPR resulting in a robust increase of the hPR promoter activity. PMID- 12088867 TI - Evidence that PACAP and GnRH down-regulate follicle-stimulating hormone-beta mRNA levels by stimulating follistatin gene expression: effects on folliculostellate cells, gonadotrophs and LbetaT2 gonadotroph cells. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) stimulates alpha subunit transcription and lengthens LH-beta mRNA transcripts, but reduces FSH beta mRNA levels in rat pituitary cell cultures. PACAP also stimulates follistatin transcription, an effect which may explain the decrease in FSH-beta mRNA. To begin to investigate the cells in which PACAP activates the follistatin gene, quantitative in situ hybridization for follistatin mRNA combined with immunostaining for LHbeta and S100 protein was performed. In control cultures, follistatin mRNA was expressed in 70% of gonadotrophs and in 47% of folliculostellate cells (S-100+). PACAP increased (P<0.001) both the number of follistatin-expressing cells as well as the number of grains per cell in both gonadotrophs and folliculostellate cells, while GnRH only affected (P=0.01) gonadotrophs. Follistatin and FSH-beta gene expression in rat pituitary cultures were also measured by competitive quantitative RT-PCR and northern analysis, respectively. Both PACAP and GnRH increased (P<0.05) follistatin gene expression and suppressed (P<0.05) FSH-beta mRNA, and the effect of PACAP together with GnRH on follistatin exceeded that of GnRH alone. PACAP regulation of follistatin and FSH-beta gene expression was studied further in LbetaT2 cells that were found to express receptors for the specific PACAP receptor, PAC(1). Follistatin mRNA was undetectable in cultures exposed to control media, or stimulated with PACAP, GnRH or rh-activin-A. In contrast to the results in primary pituitary cultures, PACAP increased FSH-beta mRNA in these follistatin-deficient cells. Moreover, using transient transfection, PACAP stimulated transcription of ovine-FSH-beta luciferase. GnRH likewise increased FSH-beta mRNA and stimulated FSH-beta gene transcription in LbetaT2 cells. Activin-A increased FSH-beta gene expression dose dependently, and activin induction of FSH-beta mRNA was blocked completely by 3 fold excess follistatin. These results indicate that PACAP stimulates follistatin gene expression in both gonadotrophs and folliculostellate cells, and provide further evidence that follistatin is required for PACAP or continuous GnRH to down-regulate FSH-beta mRNA. These experiments suggest a mechanism by which PACAP influences FSH production selectively by an autocrine effect on gonadotrophs and by a paracrine mechanism through folliculostellate cells that involves follistatin. PMID- 12088868 TI - G120K-PEG, a human GH antagonist, decreases GH signal transduction in the liver of mice. AB - After receptor binding, growth hormone (GH) induces GH receptors (GHR) dimerization and JAK2 is activated after its association with a dimerized GHR, stimulating the tyrosyl phosphorylation of insulin receptor substrate-1 (IRS-1), IRS-2 and Shc proteins. G120K-PEG, a GH antagonist is produced by a mutation that blocks GH action by preventing the GHR dimerization. This study shows that the inhibitory effect of G120K-PEG was maximal with a GH:G120K-PEG ratio of 1:100, as no increase in JAK2 tyrosyl phosphorylation was observed with this dose of GH. When the dose of GH was increased and with a GH:G120K-PEG ratio of 1:10 some tyrosyl phosphorylation of JAK2 could be observed. Additionally, GH-induced IRS 1, IRS-2 and SHC tyrosyl phosphorylation was inhibited approximately 50% at equimolar concentrations of the antagonist of GH and almost abolished with a GH:G120K-PEG ratio of 1:100. The results clearly show that G120K-PEG inhibits GH signal transduction in mouse liver. PMID- 12088869 TI - Hyperglycemia enhances VSMC proliferation with NF-kappaB activation by angiotensin II and E2F-1 augmentation by growth factors. AB - To clarify the mechanisms of hyperglycemia-induced proliferation of vascular smooth muscle cells (VSMC), we examined the effects of high glucose (HG) on nuclear factor (NF)-kappaB and E2F-1. Angiotensin II (Ang II) significantly enhanced DNA binding activity of NF-kappaB under HG (25.6 mM) conditions with an increase in p65 subunit of NF-kappaB, and did it slightly under normal glucose (NG; 5.6 mM) conditions. Ang II failed to induce E2F-1 expression, or its binding to the cdc2 promoter, even under HG conditions. HG greatly augmented the cdc2 inducibility of fetal calf serum (FCS), through the increase in E2F-1 activity. These data indicate that hyperglycemia contributes to abnormal proliferation of VSMC by two mechanisms; the induction of NF-kappaB activation by Ang II, which facilitates transcription of certain growth factors, and the augmentation of E2F 1 in response to growth factors. PMID- 12088870 TI - The zinc finger/homeodomain protein deltaEF1 mediates estrogen-specific induction of the ovalbumin gene. AB - Regulation of the ovalbumin (Ov) gene is strictly controlled by precise developmental, tissue-specific, and hormonal cues. The Ov gene is transcriptionally activated by four classes of steroid hormones: estrogens, androgens, glucocorticoids, and progestins. Although it has served as a model to study multi-hormone gene regulation for the past 30 years, the pathways that relay each hormone signal to the Ov gene are largely unclear. Extensive linker scanner and point mutation analysis has revealed elements necessary for its induction by estrogen, androgen, progesterone, or glucocorticoid but has failed to identify any elements that are specific to the action of any one steroid hormone. These observations in conjunction with the observation that the Ov gene is indirectly regulated by steroid hormones suggest that these signals may all induce the same transcription factor. However, here we have identified two cis acting DNA elements in the 5' flanking region of the Ov gene that are required for induction by estrogen, but not by androgen or progesterone. These elements span -152 to -146 and -810 to -806 with respect to the start point of transcription. This implies that estrogen induces the Ov gene by a separate pathway than do androgens or progestins. Gel mobility shift assays demonstrate that the estrogen-specific sequences bind the estrogen inducible transcription factor deltaEF1, suggesting that deltaEF1 plays a distinct role in mediating the estrogen signal to the Ov gene. PMID- 12088871 TI - Estrogen receptor-mediated effects of tamoxifen on human endometrial cancer cells. AB - Tamoxifen is an estrogen receptor (ER)-antagonist that is widely used for the treatment of breast cancer, although it increases the risk of endometrial cancer. The mechanism mediating the stimulatory effect of tamoxifen on endometrial cancer is presently unknown. In this study we examined the effects of tamoxifen on Ishikawa 3H-12 endometrial cancer cells and MCF-7 breast cancer cells. Ishikawa cell growth was stimulated by 4-hydroxytamoxifen and accompanied by increased transcriptional activity of the endogenous ER. These stimulatory effects did not occur in MCF-7 cells. The relative transcriptional activity of the activation function (AF) 1 domain of ERalpha compared with that of the AF2 domain was 4-fold higher in Ishikawa cells than in MCF-7 cells. Mitogen-activated protein (MAP) kinase, which stimulates the transcriptional activity of AF1, was constitutively activated in Ishikawa cells, but not in MCF-7 cells. These observations suggest that the constitutively activated MAP kinase-signaling pathway in Ishikawa cells enhances the transcriptional activity of ERalpha via the AF1 domain. This ERalpha activation pathway may be involved in the stimulatory effect of tamoxifen on the development and/or progression of endometrial cancer. PMID- 12088872 TI - Expression and modification of PKA and AKAPs during meiosis in rat oocytes. AB - Meiosis in oocytes is initiated during fetal life, arrested around birth and resumed after puberty. Meiotic arrest is controlled by a cAMP-dependent protein kinase (PKA)-mediated cAMP action. We examined oocytes for the presence and modulation of the regulatory (R) subunits of PKA and the A-kinase anchoring proteins (AKAPs) that target PKA to specific subcellular locations. We found that rat oocytes express the two regulatory subunit isoforms, RI and RII of PKA. Immunocytochemistry revealed that the regulatory subunits underwent cellular translocation upon resumption of meiosis. We also demonstrated the presence of a novel 140 kDa AKAP, AKAP140 that exhibited a retarded electrophoretic motility at reinitiation of meiosis. The mobility shift of AKAP140 was susceptible to alkaline phosphatase and prevented by inhibition of p34cdc2 kinase. We conclude that rat oocytes express AKAP140 that is phosphorylated during meiosis. AKAP140 phosphorylation is sensitive to p34cdc2 kinase inhibitors. We hypothesize that AKAP140 and its phosphorylation state may influence the translocation of the R subunits of PKA throughout resumption of meiosis. PMID- 12088873 TI - Expression of mRNA encoding growth differentiation factor 9 and bone morphogenetic protein 15 during follicular formation and growth in a marsupial, the brushtail possum (Trichosurus vulpecula). AB - The oocyte derived growth differentiation factor (GDF) 9 and bone morphogenetic protein 15 (BMP15; also known as GDF9b) are essential for normal follicular growth. However, little is known about expression of these factors during ovarian development. Therefore, we determined the ontogeny of expression of GDF9 and BMP15 mRNA in the developing ovary of the brushtail possum. Ovaries were collected from pouch young (n=3-5 per group) around times of key developmental events namely: (1) morphological sexual differentiation (i.e. days 1-5 following birth), (2) after sexual differentiation (i.e. days 10-15), (3) before and during initiation of germ-cell meiosis (i.e. days 22-45), (4) shortly after initiation of follicular growth (i.e. days 78-85), (5) during preantral follicular growth (i.e. days 96-113) and (6) during antral follicular growth (i.e. days 155-190). Ovaries were also collected from three juvenile and four adult animals and gene expression was determined by in situ hybridization. The mRNAs encoding GDF9 and BMP15 were first observed in oocytes of newly-formed primordial follicles (i.e. days 78-85). Expression of both mRNAs was restricted to the oocyte and was present in follicles irrespective of whether they were non-growing primordial follicles or undergoing preantral or antral development. Thus, since the mRNAs encoding GDF9 and BMP15 were not observed until follicular formation, it is unlikely that these proteins have any role in early germ cell development. Nevertheless, the findings that the mRNAs encoding both proteins were observed in oocytes from the primordial stage of follicular formation suggest a possible role for these proteins in the maintenance of primordial follicles as well as a key role during follicular development. These results highlight important species differences in the ontogeny of expression of GDF9 and BMP15 between possums and other species such as the human, sheep or rat. PMID- 12088874 TI - Hormonal regulation of DNA polymerase beta activity and expression in rat adrenal glands and testes. AB - We investigated whether DNA polymerase beta activity and expression in rat adrenal glands and testes are controlled by the cAMP dependent protein kinase (A kinase) phosphorylation system in addition to anterior pituitary hormones. DNA polymerase beta mRNA expression in rat testes was decreased by hypophysectomy and recovered with administration of gonadotropic hormone, suggesting that this enzyme is controlled at the mRNA level by this pituitary hormone. In addition, DNA polymerase beta activity in the adrenal glands and testes and the amount of mRNA in the testes increased when cAMP was administered to the normal rat. This activity was decreased by administration of the cyclic AMP-dependent protein kinase inhibitor, H(8). Moreover, when alkaline phosphatase was added to the assay system in vitro, a decrease in DNA polymerase beta activity was observed. These findings indicate that changes in the activity and expression of DNA polymerase beta are mediated via cAMP and the A-kinase system, and that phosphorylation of this enzyme is also involved in this expression. PMID- 12088875 TI - Mouse pituitary adenylate cyclase-activating polypeptide (PACAP): gene, expression and novel splicing. AB - PACAP is a conserved neuropeptide present in all vertebrates. In the mouse, the PACAP gene and various mRNAs have been isolated. To further characterize the mouse PACAP gene (Adcyap1), we have confirmed its sequence, identified its chromosomal location on distal chromosome 17 and used RT-PCR and 5'RACE in various tissues to identify eight PACAP mRNAs, four of which are novel. Three of these novel transcripts have alternate 5'UTRs, whereas the fourth is altered within the coding region. This is the first report of alternate splicing within the coding region of the PACAP gene. The expression pattern of PACAP in the mouse during embryonic development and adulthood is known. Here, expression of PACAP in the mouse at four postnatal stages in 12 tissues is assessed. We identify continuous expression of PACAP in the brain and eye and stage-specific expression in the gonads and thymus. The complex splicing of PACAP transcripts may regulate the tissue-and stage-specific expression. PMID- 12088876 TI - MT(1) melatonin receptor overexpression enhances the growth suppressive effect of melatonin in human breast cancer cells. AB - Melatonin inhibits the proliferation of estrogen receptor alpha (ERalpha) positive (MCF-7), but not ERalpha-negative (MDA-MB-231) breast cancer cells. Here, we assessed the effect of MT(1) melatonin receptor stable overexpression in MCF-7 and MDA-MB-231 breast cancer cells on the growth-suppressive effects of melatonin. Parental and vector-transfected MCF-7 cells demonstrated a modest, but significant, growth-suppressive response to melatonin; however, melatonin treatment of MT(1)-transfected MCF-7 cells resulted in significantly enhanced growth-suppression. This response was blocked by an MT1/MT2 melatonin receptor antagonist. Interestingly, MT(1)-overexpression did not induce a melatonin sensitive phenotype in melatonin-insensitive MDA-MB-231 cells. Finally, Northern blot analysis demonstrated an enhanced inhibition of ERalpha mRNA expression and an enhanced induction of pancreatic spasmolytic polypeptide (pS2) by melatonin in MT(1)-transfected MCF-7 cells relative to vector-transfected MCF-7 cells. These data suggest the involvement of the MT(1) melatonin receptor in mediation of melatonin effects on growth-suppression and gene-modulation in breast cancer cells. PMID- 12088877 TI - Organochlorine pesticides directly regulate gonadotropin-releasing hormone gene expression and biosynthesis in the GT1-7 hypothalamic cell line. AB - Environmental toxicants profoundly affect growth and developmental processes. In the present study, we hypothesized that hypothalamic gonadotropin-releasing hormone (GnRH) neurons, which regulate the reproductive axis, are targets of environmental endocrine disrupting chemicals. Two organochlorine pesticides (methoxychlor and chlorpyrifos) were tested for their effects on GnRH gene expression and biosynthesis in the immortalized hypothalamic GT1-7 cells, which synthesize and secrete GnRH. GT1-7 cells were treated with methoxychlor or chlorpyrifos for 24 h in dose-response experiments, and GnRH gene expression and peptide levels were quantified. In order to examine whether these pesticides affect GnRH biosynthesis through the estrogen receptor (ER), in other experiments their effects were compared to those of estrogen, or they were co-administered with the ER antagonist, ICI 182,780 (ICI). Both methoxychlor and chlorpyrifos had significant effects on GnRH gene transcription and GnRH mRNA levels. These effects were not consistently blocked by ICI, nor did the effects of these pesticides consistently mimic those of estrogen, suggesting a mechanism independent of the ER. Chlorpyrifos and methoxychlor slightly stimulated peptide levels, and this effect was blocked by ICI, suggesting that the ER may mediate effects of pesticides on GnRH release. These results indicate that chlorpyrifos and methoxychlor alter GnRH biosynthesis in this hypothalamic cell line in vitro, suggesting that they may have endocrine disrupting effects on GnRH neurons in vivo. PMID- 12088878 TI - Structural regions of ERalpha critical for synergistic transcriptional responses contain co-factor interacting surfaces. AB - Most highly estrogen responsive genes are synergistically activated by multiple copies of estrogen responsive elements (EREs) capable of binding to the estrogen receptor (ER). We examined here the structural features of the receptor necessary to interact with co-regulatory proteins and to produce a synergistic pattern of activation from multiple EREs. Using full length and truncated variants of ERalpha, we show in transfected mammalian cells that although the carboxyl (AF-2) and the amino (AF-1) terminal activation domains are functionally integrated to induce transcription, AF-1 is critical for mediating synergy. Partial characterization of AF-1 sub-domains revealed that both Box-1 and Box-2 regions (amino acids 41-64 and 87-108, respectively) are essential for a synergistic response to estrogen. We show that members of the p160 family of co-factors and TIF-1 interact with the AF-2 domain of ERalpha. We also found that TIF-2, a member of the p160 family, can interact with the Box-1 region of AF-1. Apparently, structural regions required for the ability of ERalpha to induce transcription synergistically from tandem ERE sequences are also critical for the interaction of the receptor with the co-regulatory proteins. PMID- 12088879 TI - Regulation of endometrial adenocarcinoma cell proliferation by Activin-A and its modulation by 17beta-estradiol. AB - A role for activins in regulating cellular transformation is suggested by the alpha-inhibin knockout mouse in which development of gonadal tumors is associated with elevated activin levels. It was the purpose of the current study to determine whether activin had similar actions on endometrial cell lines, specifically on a well differentiated estrogen-responsive endometrial adenocarcinoma cell line (ISH) and estrogen-unresponsive cells (HEC-50) obtained from a poorly differentiated endometrial adenocarcinoma. Activin was secreted by both adenocarcinoma cell lines. Using reverse transcription-PCR, messenger RNA type I and type II activin receptor subtypes were detected in both cell lines: expression of IB and IIB was approximately three- to fourfold greater in ISH cells than in HEC-50 cells, while activin receptor IA and IIA messenger RNA levels were approximately equal in both cell lines. Activin treatment (30-300 ng/ml) caused a dose- and time-dependent inhibition of ISH cells proliferation and resulted in a significant decrease in Bcl-2 protein and mRNA levels. No difference was observed in Bax expression. There was no significant effect of activin when the cultures of ISH cells were exposed to 17beta-estradiol. In contrast, activin showed a weak, but significant, mitogenic effect on HEC-50 cells without modifications in Bax and Bcl-2 mRNA and protein levels. The results demonstrate that activin is a regulator of endometrial cancer cell growth. 17beta Estradiol may promote resistance of estrogen-responsive endometrial cancer cells to the growth-retarding effects of activin and one of the mechanisms might be a down-regulation of the activin receptors. PMID- 12088880 TI - Hypoxia decreases Runx2/Cbfa1 expression in human osteoblast-like cells. AB - To elucidate the molecular mechanism in relation to vascular supply and osteoporosis, we investigated the effect of hypoxia on Runx2 expression in MG63 cells. Also investigated was expression of type I collagen and osteocalcin, which are regulated by Runx2, alkaline phosphatase (ALPase) to see if they are affected by hypoxia. Quiescent cultures of MG63 cells were exposed to hypoxia (2% O(2)) and normoxia (18% O(2)) for 24, 48, 72 and 96 h. In cells exposed to hypoxia, reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that mRNA expression of Runx2, type I collagen, osteocalcin, and ALPase were decreased in a time dependent manner to 96 h. Activity of ALPase was also reduced in the same manner. Western blotting showed a marked decrease in Runx2 protein at 96 h in cells under hypoxia compared to normoxia. These data indicate that Runx2 expression in osteoblasts is reduced by hypoxia, and may be a mechanism of osteoporosis by decreased vascular supply. PMID- 12088881 TI - Affinity chromatography and co-chromatography of bispecific monoclonal antibody immunoconjugates. AB - Bispecific monoclonal antibodies (bsMAb) are unique macromolecules functioning as cross-linkers with two different predetermined binding specificities. A wide range of potential applications employing these probes can be envisioned in immunodiagnostics and immunotherapy. One of the major limitations for the use of bsMAbs produced by hybrid-hybridomas is the production of parental monospecific antibodies along with bsMAbs. Hence, the purification of desired bsMAb free from both parental mAbs and other possible promiscuous combinations is essential. Purification of antibodies is the single greatest obstacle in obtaining an immunoprobe with high specific activity. This review describes the affinity purification and affinity co-purification techniques for the separation of bsMAb as a pre-formed immune complex or as a pure species. The use of immobilized ligands is the basis of affinity chromatography. Affinity chromatography can be classified into three different categories depending on the properties of the immobilized ligand. The ligand-specific affinity chromatography is based on the extremely specific immobilized ligand, directed towards the protein or antibody of interest. Using a dual, sequential affinity chromatography, bsMAb can be purified from a mixture of bispecific and monospecific monoclonal antibodies with a ligand specific for each antibody. Thiophilic adsorption is a group-specific affinity method that can be successfully used to separate monospecific forms from bispecific species by salt gradient elution. Affinity co-chromatography offers a convenient one-step method for purification of bulk amounts of immunoconjugates for diagnostic applications by exploiting several dye-ligands known to bind certain enzymes. The same method could be potentially used for quality control and quality assurance purposes in industrial biotechnology. PMID- 12088882 TI - Purification of antibodies by affinity chromatography. AB - This review focusses on affinity purification of immunoglobulins, a methodology which is a powerful tool to obtain pure and intact antibodies. Affinity techniques allow antibody purification both in a single step chromatographic procedure as well as in complex purification protocols depending on the intention to use the target antibody. The purification strategies for antibodies by interaction with affinity ligands such as antibodies and Fe receptors or low molecular weight compounds are described. PMID- 12088883 TI - Mutation detection of the human glucocorticoid receptor alpha gene area coding for the hormone-binding domain by denaturing gradient gel electrophoresis. AB - Mutations in the hormone-binding domain of the human glucocorticoid receptor alpha (hGRalpha) gene have been detected in a variety of glucocorticoid resistance syndromes. Using the denaturing gradient gel electrophoresis technique, we developed a sensitive method for the detection of alterations in the gene area coding for the whole hormone-binding domain and part of the DNA binding domain of the hGRalpha. This method can be applied for screening of glucocorticoid receptor gene alterations in glucocorticoid-dependent diseases. PMID- 12088884 TI - Monitoring and control of pullulan production using vision sensor. AB - The production of the polysaccharide pullulan by the yeast-like fungi, Aureobasidium pullulans, is accompained by cellular morphogenetic changes. High productivity and yield of the process have been found to correlate with high concentration of yeast-like cells in the culture. The morphogenetic changes of A. pullulans cells depend on the culture conditions, e.g., dissolved oxygen, shear rate and medium composition. In order to improve the productivity of the process, a novel control law was formulated. A feeding strategy dependent on the culture cellular composition was designed and aimed to keep the yeast-like cell concentration high. The culture morphogenetic composition during the process was monitored by a recently developed vision sensor. Feeding was actuated when the yeast-like cell concentration decreased below a threshold. The proposed control strategy improved pullulan production by increasing both productivity and yield of the cells by 67% and 80%, correspondingly. The results point to the advantage and the potential of using the monitoring and control system and algorithm to increase productivity and yield in cellular bioprocesses. PMID- 12088885 TI - Novel methodology for the follow-up of acute lymphoblastic leukemia using FTIR microspectroscopy. AB - In this report, we present a novel spectroscopic method of follow-up during chemotherapy treatment for B- and T-cell childhood leukemia patients. We isolated peripheral lymphocytes from blood drawn from patients before and after the chemotherapy and collected Microscopic FTIR (FTIR-MC) spectra of the isolated lymphocytes. Our results showed that nucleic acids content decreased in both types of patients. Changes in phospholipids and proteins level could be observed. The overall effects of drugs administered to the patients can be understood at the molecular level using FTIR-MC and these results are expected to stimulate wider applications of spectroscopy in leukemia research. PMID- 12088886 TI - Determination of glucose oxidase immobilised as monolayer onto a flat surface. AB - An assay to estimate the amount of glucose oxidase immobilised as a monolayer onto a flat surface is reported. This method is based on the electrochemical detection of the flavin adenine dinucleotide (FAD) cofactor released by the immobilised enzyme in acid solutions. FAD concentration in the acid solution was measured by amperometry, using a flow injection analysis (FIA) system equipped with a wall-jet electrode, and with a sensitivity of (9.2+/-2.0)x10(-2) nA/nM. By this method, the amount of glucose oxidase molecules present in a monolayer deposited on a silanised glass slide was easily detected, in which the detection limit is more than one order of magnitude lower than the maximum loading of the surface with an ordered monolayer of glucose oxidase. PMID- 12088887 TI - 2-D gel densitometer for high-contrast and selective imaging of chlorophyll containing protein complexes separated by non-denaturing polyacrylamide gel electrophoresis. AB - In this work, we present a home-made two-dimensional (2-D) CCD imaging system for the monochromatic densitometry of plane gels and its application to the imaging and densitometry of chlorophyll (Chl)-containing proteins separated by non denaturing polyacrylamide gel electrophoresis. The monochromatic imaging of separated green bands at the wavelengths corresponding to their absorption band increases their contrast. This allows a better visualization of the faint-green bands in the gel and using of samples with lower Chl content for the electrophoresis. By the comparison of 2-D densitograms of the same gel illuminated with 670 and 650 nm lights, that is, at the red absorption maximum of Chl a and b, respectively, we achieved a selective imaging of the complexes with different Chl a/b ratio. This approach was used to specify an unknown band that appeared in the gel of the sample prepared from the thylakoid membranes of preheated barley leaves. PMID- 12088888 TI - Social and political factors influencing the functioning of regional health boards in British Columbia (Canada). AB - Health reform is associated with changes in the way the health system works and in the roles of major stakeholders, such as governments, health professionals, and the lay public. This paper reviews the immediate relevance of these social and political elements to health boards, particularly those with lay board members; source documents include peer-reviewed articles, and government documents and news releases in Canada especially. Also presented are the perceptions of 130 regional health board members in British Columbia (BC), Canada, who responded to our 1996 survey questionnaire. Two sets of social and political factors are identified and discussed in this paper. The first set deals with the composition of health board members (qualifications, representation, and selection). Our findings suggest that there is now less attention focusing on the composition of health boards in BC. This may contribute to a re-focusing of attention on the boards' effectiveness in working with stakeholders and in influencing the health system. The other set of social and political factors deals with the relations of health boards with key stakeholder groups. The responses to our questionnaire suggest that the health boards in BC may have had some success in addressing the concerns of various stakeholder groups. However, the respondents also suggested that the stakeholder groups needed to be more understanding and involved in the regionalization (decentralization) process. Health boards that have lay representatives, including regional health authorities in Canada, face similar social and political factors immediate to their operation. PMID- 12088889 TI - A follow-up study of drug misusers who received an intervention from a local arrest referral scheme. AB - In the United Kingdom the election of the 1997-2001 Labour government led to rapid development of drug treatment interventions in the criminal justice sector. One type of drug treatment intervention is arrest referral and this is the most developed and researched form of intervention within the United Kingdom (Arrest referral: emerging lessons from research, paper 23. London: Home Office; 1998). However, much of the published literature reports upon evaluations of working styles and practices of individual arrest referral schemes (ARS). This paper reports a follow-up study on the impact that intervention by an ARS had upon a group of drug-misusing arrestees, 18 months after their initial contact with the ARS. The findings from the follow-up study demonstrate that the intervention had had a positive impact on both drug misuse and offending among the sample. Sixty four per cent had entered treatment and remained 'clean' following an intervention by the ARS. Analysis of the police national computer records for the sample demonstrated that 88% of those who remained 'clean' also ceased criminal activity. Additionally, the study highlighted other factors relevant to the treatment of drug misusers. Most of the sample had been in contact with health professionals during their misusing careers but the perceived negative attitude on the part of health professionals towards drug misusers and the lack of proactive, health-led interventions indicated that mainstream health care provision had failed this sample of problematic drug misusers. Another key factor to emerge from the study was the relevance of aftercare to the recovery process, which was needed to offer wide-ranging support to drug misusers in recovery, such as help with accommodation, learning basic life skills and constructing a new lifestyle without drugs. The study therefore has a dual outcome in demonstrating the positive impact of the ARS and in turn the rationale for the Government's 10 year anti-drugs strategy but it also highlights the weakness as aftercare is not a prominent feature of the strategy (Tackling drugs to build a better Britain, the government's ten year strategy for tackling drugs. London: The Stationery Office; 1998). PMID- 12088890 TI - Productivity costs before and after absence from work: as important as common? AB - Traditionally, production losses are estimated using the human capital or friction cost method. These methods base estimations of productivity costs on data on absence from work. For some diseases, like migraine, productivity losses without absence are occasionally calculated by estimating the production losses from reduced productivity at work. However, diseases typically only associated with absence may also be expected to cause reduced productivity before and after absence. In a previous study, Brouwer et al. concluded that productivity losses without absence are also very relevant in common diseases, like influenza, common cold or neck-problems. Studying a new sample of employees of a Dutch trade-firm (n = 51), who completed the questionnaire 'Ill and Recovered' upon return to work after absence due to illness, it was revealed that about 25% of the respondents experienced production losses before absence and about 20% of the respondents experience production losses after absence. This leads to an increase in estimated production losses of about 16% compared with only considering absence data. Current productivity costs estimates based solely on absence data may, therefore, underestimate real productivity costs. Compensation mechanisms in firms may reduce the underestimation. PMID- 12088891 TI - Exploring the role of order effects in person trade-off elicitations. AB - BACKGROUND: The person trade-off (PTO) has been advocated by some as an alternative measure for the purposes of cost-effectiveness analyses. However, the measurement properties of PTO elicitations are still being defined. METHODS: We presented subjects with two PTO scenarios. In the pre-existing paraplegia scenario, they were asked how many paraplegics' lives would have to be saved to be just as important as saving 100 'normal' people's lives. In the paraplegia onset scenario, they were asked how many patients who would experience the onset of paraplegia need to be saved to equal the benefit of saving 100 'normal' lives. We varied the order of the two scenarios across subjects to test whether PTO elicitations are susceptible to order effects. In addition, we varied whether subjects were required to provide a numerical response to the first elicitation. RESULTS: Subjects' PTO indifference points for the two scenarios varied dramatically depending on the order with which they received the scenarios, and according to whether the first elicitation required a numerical response. For those subjects providing numerical responses to both elicitations, median PTO responses varied by a factor of close to two in the pre-existing paraplegia scenario and by a factor of eight in the paraplegia onset scenario. However, the magnitude of the order effect was significantly reduced when subjects were not asked to provide a numerical response to the first PTO elicitation. CONCLUSION: PTO elicitations are susceptible to order effects. These order effects are partly due to numerical anchoring. However, other cognitive factors contribute to the order effect. Further research should clarify whether these order effects can be reduced. PMID- 12088892 TI - A combination of an extrapolation method and a benchmark method to develop quantitative health targets for Japan. AB - The aim of the present study was to propose a methodology to formulate quantitative health targets which combined an extrapolation method and a benchmark method, and to estimate the targets for mortality rates (Mb) for selected causes of death by the year 2010 in Japan. Using the extrapolation method, based on the nationwide Mt from 1988 to 1997, the Mt in 2010 was predicted using a regression model. Using the benchmark method, the paired-mean of the age-adjusted mortality rates (Mts) of the top five prefectures among 47 prefectures was calculated as the benchmark. Combining the predicted mortality and the benchmark mortality, year 2010 targets were determined. As a results year 2010 targets as percentages compared with Mt in 1997 for cancer at all sites, stomach cancer, lung cancer, colo-rectal cancer, liver cancer and stroke were estimated to be 93, 52, 94, 102, 53 and 52% for males, and 84, 43, 86, 82, 60, and 45%, for females, respectively. The methodology presented in this article could be used as a standard procedure to formulate realistic quantified health targets, which can be adopted to develop health policies in nations, regions and communities. PMID- 12088893 TI - Consumer involvement in health research: a review and research agenda. AB - The involvement of consumers in health research is now Department of Health policy within the UK. Despite the existence of policy directives, there is a dearth of knowledge on the effects of such involvement. This paper critically reviews the state of our knowledge on this issue, and maps out a research agenda with the aim of stimulating systematic, empirical inquiry into consumer involvement in health research. The paper discusses definitions of 'the consumer'; considers why consumer involvement is believed to be important to health research; traces the development of the policy; analyses the epistemological and methodological implications of the policy; discusses the various levels of consumer involvement in research; and outlines the objections to the policy that have been put forward by clinicians and researchers. Four questions were identified during the review as being in need of theoretical and empirical attention: (1) how can consumer involvement in health research be further conceptualised? (2) how and why does consumer involvement influence health research? (3) how can the influence of consumers in health research be measured and evaluated? and (4) what factors are associated with 'successful' consumer involvement in health research? PMID- 12088914 TI - Controversies in radioiodine therapy: relation to ophthalmopathy, the possible radioprotective effect of antithyroid drugs, and use in large goitres. AB - In routine use for more than 50 years, radioiodine ((131)I) is generally considered safe and devoid of major side effects. Therefore, it is surprising that relatively many aspects of radioiodine therapy are controversial, as illustrated by recent international questionnaire studies. Our review aims at highlighting three of these areas - namely, the influence of (131)I on the course of Graves' ophthalmopathy, the possible radioprotective effects of antithyroid drugs, and the use of (131)I in large goitres. (131)I therapy carries a small (but definite) risk of causing progression of Graves' ophthalmopathy. Identification of risk factors (thyroid dysfunction, high level of thyroid stimulating hormone (TSH) receptor antibodies, cigarette smoking) allows the identification of patients at risk and the institution of concomitant glucocorticoid treatment, thereby hindering progression of eye disease. On the basis, largely, of retrospective data, it appears that carbimazole (or methimazole), if stopped 3-5 days before treatment, does not influence the outcome of (131)I therapy. Simultaneous thyrostatic medication most probably reduces the efficacy of (131)I, as does restarting it within 7 days. Propylthiouracil seems to have a more prolonged radioprotective effect than carbimazole. Surgery is the treatment of first choice in patients with a large goitre. However, in the case of patient ineligibility or preference, (131)I therapy may be an option. The treatment has a favourable effect on tracheal compression and inspiratory capacity, but the reduction in thyroid volume is only 30-40%. Inpatient treatment, necessitated by the large doses, makes the treatment cumbersome. Controversy related to radioiodine therapy is mainly based on the lack of adequate prospective randomised studies comparing efficacy, side effects, cost and patient satisfaction. PMID- 12088915 TI - Functional scintigraphy of the adrenal gland. AB - Over the last 30 years nuclear medicine imaging of the adrenal gland and its lesions has been achieved by the exploitation of a number of physiological characteristics of this organ. By seeking and utilising features which are quantitatively or qualitatively different from those of the adjacent tissues, functional depiction of the adrenal gland and its diseases, which in most cases retain the basic physiology of their tissue of origin, including both the cortex and the medulla, are now a useful clinical reality. Agents widely used in clinical practice include: (a) uptake and storage of radiolabelled cholesterol analogues via the low density lipoprotein (LDL) receptor and cholesterol ester storage pool in the adrenal cortex ((131)I-6-beta-iodomethyl-norcholesterol, (75)Se-selenomethyl-norcholesterol); (b) catecholamine type I, presynaptic, uptake mechanism and intracellular granule uptake and storage mechanism in the adrenal medulla and extra-adrenal paraganglia ((131)I-, (123)I- and (124)I-meta iodo-benzyl-guanidine (MIBG), (18)F-metafluoro-benzyl-guanidine); (c) cell surface receptor binding of peptides/neurotransmitters/modulators such as for the family of five subtypes of somatostatin receptors ((123)I-tyr-octreotide, (111)In DTPA-octreotide, (111)In-DOTA-octreotide and many others); (d) although not specific for the adrenal gland, increased glycolysis by tumours, particularly the most malignant varieties, (18)F-2-fluoro-d-deoxyglucose can thus be expected to depict certain malignant lesions such as malignant pheochromocytomas (particularly the minority which are not detected by MIBG) and adrenal incidentalomas (particularly when they occur in patients with known extra-adrenal malignancies). There are a variety of adrenal tissue characteristics with potential for exploitation but which are not currently in clinical use, and which may, nevertheless, have potential as imaging agents. These include: (a) inhibitors of adrenal cortical steroid hormone synthesis enzymes (e.g. radiolabelled analogues of metyrapone); (b) radiolabelled lipoproteins which bind to adrenocortical LDL receptors; (c) inhibitors of catecholamine biosynthesis enzymes (e.g. radiolabelled analogues of tyrosine and related amino acids); (d) cell surface receptors for various peptides and hormones which may be over expressed on adrenal cortical or adrenal medullary tumours (e.g. radiolabelled analogues of ACTH on adrenocortical cells of zona fasciculata or zona glomerulosa origin, neurotransmitter/hormone message peptides binding to cell surface receptors such as bombesin, vasoactive intestinal polypeptide, cholecystokinin and opiate peptides); (e) the adrenal cortex can also synthesise cholesterol ab initio from acetate, and preliminary studies with (11)C-acetate positron emission tomography have shown interesting results. PMID- 12088916 TI - Comparison of two different doses of iodide in the prevention of gestational goiter in marginal iodine deficiency: a longitudinal study. AB - OBJECTIVE: A prospective randomized trial was performed to assess the usefulness of iodine supplementation in the prevention of goiter in pregnant women living in marginally iodine-deficient areas. DESIGN: Eighty-six pregnant women were recruited and randomized in two groups and treated daily for up to six months after delivery with 200 microg iodide (group A) or 50 microg iodide (group B). Sixty-seven women (32 in group A and 35 in group B) completed the study. METHODS: Thyroid volume (TV), thyroid functional parameters and urinary iodine concentration were determined in all subjects at booking, at the 18th-26th, and the 29th-33rd week of gestation, and at the 3rd and 6th month after delivery. RESULTS: A slight but not significant increase in TV during gestation was observed only in group B. After delivery a progressive decrease in TV was documented in both groups, the final TV being significantly reduced with respect to the initial volume in group A. No significant changes in serum free thyroid hormones and TSH concentrations were found during gestation in either group. Postpartum thyroiditis was observed in 5 women (2 in group A, 3 in group B). No side effects were seen. CONCLUSION: The present data indicate that in marginally iodine-deficient areas, the administration of iodide is recommended in pregnancy and lactation. In the conditions of the present trial a dose of 50 microg iodide/day is a safe and effective measure in preventing an increase in TV during pregnancy but a dose of 200 microg iodide/day appeared to be more effective without inducing side effects and without enhancing the frequency of post-partum thyroiditis. PMID- 12088917 TI - The effects of thyrotoxicosis and its treatment on central arterial stiffness. AB - OBJECTIVE: To assess central arterial stiffness in thyrotoxicosis using the technique of pulse wave analysis. DESIGN: Case control study designed to determine the effect of thyrotoxicosis on central arterial stiffness and at 6 months after radioiodine treatment. PATIENTS: Twenty (18 women and 2 men) thyrotoxic patients and 20 age- and sex-matched controls were studied at baseline. Thyrotoxic patients were re-studied at 6 months following treatment of thyrotoxicosis with 555 MBq (131)I with no additional therapy for the six-month period. MEASUREMENTS: Using the sphygmocor apparatus, peripheral pressure waveforms were recorded non-invasively from the radial artery and central pressure waveforms were generated from these. Indices of arterial stiffness, central augmentation index (AI), augmentation of central arterial pressure (AG) and central blood pressures were derived. AI corrected for heart rate (AIc) was calculated. RESULTS: Thyrotoxic patients recorded a significantly lower AI (means+/-s.e.m.) compared with controls (15.0+/-2.1 vs 28.0+/-2.1%; P<0.0005) even when corrected for differences in heart rate AIc (20.0+/-2.1 vs 28.0+/-2.1%; P<0.005) as well as AG (6.0+/-0.8 vs 10.0+/-1.1 mmHg; P<0.002) but higher pulse pressure (58.0+/-3.5 vs 47.0+/-2.0 mmHg; P<0.02). At 6 months following treatment, a significant rise in AIc (27.0+/-1.8 vs 20.0+/-2.1%; P<0.005) and AG (11.0+/-1.0 vs 6.0+/-0.8 mmHg; P<0.005) was noted. Lipid profiles were comparable between the groups. CONCLUSIONS: These data suggested that subjects with untreated thyrotoxicosis have a decreased augmentation of central arterial pressure or lowered central arterial stiffness that would not appear to contribute to any excess cardiovascular risk in that condition. PMID- 12088918 TI - An audit of the insulin-tolerance test in 255 patients with pituitary disease. AB - OBJECTIVE: The insulin-tolerance test (ITT) is currently considered to be the gold standard for evaluating adults suspected of GH deficiency (GHD). The aim of this study was to determine factors that may influence nadir blood glucose (BG) when using a mean insulin dose of 0.1 IU/kg body weight. Furthermore, we wanted to evaluate the safety and GH-related aspects of the ITT. DESIGN: ITT was performed in 277 patients, of whom 255 (129 females) were eligible for evaluation. RESULTS: Multiple regression analysis, including the whole population, showed that the major determining factors for nadir BG were basal BG and body mass index (BMI) (P<0.02). No serious adverse event was recorded. Sixty three percent of all patients tested had severe GHD with peak GH response to hypoglycaemia below 7.8 mIU/l. The positive predictive value for IGF-I was 0.82 and the negative predictive value was 0.47, using a cut-off value corresponding to -2 s.d. GH peak response to hypoglycaemia decreased with increasing numbers of other pituitary hormone deficiencies. CONCLUSIONS: When determining the dose of insulin based on weight, factors like pre-test BG and BMI should also be considered. We propose an algorithm stating that the dose of insulin should be 0.1 IU insulin/kg body weight minus 2 IU if pre-test BG is <4.0 mmol/l and minus 2 IU if BMI is <20 kg/m(2) in order to take these factors into account. Our findings furthermore support the concept that the low-dose ITT is a safe test in adults, when performed in experienced hands. It was confirmed that IGF-I is not sufficient when diagnosing GHD in adults, and reliable stimulation tests like ITT are required in the diagnosis. PMID- 12088919 TI - A dose titration model for recombinant GH substitution aiming at normal plasma concentrations of IGF-I in hypopituitary adults. AB - OBJECTIVE: To evaluate a dose titration model for recombinant human GH substitution in adult patients with GH deficiency, aiming at normal plasma levels of IGF-I. DESIGN AND METHODS: Eighteen patients participated and a start dose of 0.17 mg GH/day was used except by two men who started with 0.33 mg/day. To demonstrate a clear GH effect the patients were first titrated, with steps of 0.17 mg GH/day every 6-8 weeks, to IGF-I levels in the upper range of age adjusted reference values. The GH dose was then reduced 1 dose step and kept for a further 6 months. For comparison we investigated 17 healthy control subjects. RESULTS: Plasma IGF-I was increased after 2 weeks on the start dose and did not increase further for up to 8 weeks. Women had significantly lower GH sensitivity than men measured as net increment of IGF-I on the start dose of GH. GH sensitivity was not changed by age. The plasma IGF-I levels increased from 76.3+/ 47.0 (s.d.) to 237+/-97 microg/l at the end of the study (P<0.001), and similar IGF-I levels were obtained in both sexes. The maintenance median GH dose was 0.33 mg/day in males and 0.83 mg/day in females (P=0.017). The GH dose correlated negatively with age in both sexes. Body weight, very low density triglycerides, lipoprotein(a) (Lp(a)), and fasting insulin increased, whereas insulin sensitivity index (QUICKI) decreased significantly. In comparison with the controls, the patients had lower fasting blood glucose, fasting insulin and Lp(a) levels at baseline, but these differences disappeared after GH substitution. The two groups had equal insulin sensitivity (QUICKI), but 2 h oral glucose tolerance test values of blood glucose and insulin were significantly higher in the patients at the end of the study. CONCLUSIONS: In conclusion our data suggest that the starting dose of GH substitution and the dose titration steps should be individualised according to GH sensitivity (gender) and the IGF-I level aimed for (age). The reduced insulin sensitivity induced by GH substitution could be viewed as a normalisation if compared with control subjects. PMID- 12088920 TI - Growth hormone replacement does not increase serum prostate-specific antigen in hypopituitary men over 50 years. AB - OBJECTIVE: Epidemiological studies have shown an increased risk for prostate carcinoma in men with serum IGF-I in the upper part of the age-related reference range. Recombinant human GH (rhGH) is widely used in patients with GH deficiency, usually raising the serum IGF-I levels into the normal range: safety surveillance is therefore mandatory, with particular regard to neoplasia. The aim was to examine whether rhGH replacement in hypopituitary adults is associated with changes in serum prostate-specific antigen (PSA) as a surrogate marker of changes in prostatic growth. DESIGN AND METHODS: A prospective longitudinal study was used with a median follow-up of 22 (range 2.5-32) months, in which 41 men aged over 50 years with adult onset hypopituitarism and GH deficiency during rhGH replacement were examined. Serum PSA and IGF-I were measured at baseline and at latest follow-up. RESULTS: Mean serum PSA remained unchanged during rhGH replacement, with a median follow-up of 2 years. No correlation was found between the individual changes in serum IGF-I and changes in serum PSA. CONCLUSIONS: These data are reassuring thus far regarding the safety of GH replacement in relation to the prostate in this patient group. PMID- 12088921 TI - Plasma ghrelin levels during exercise in healthy subjects and in growth hormone deficient patients. AB - OBJECTIVE: To characterise plasma levels of the recently identified endogenous ligand for the GH secretagogue receptor (ghrelin) during submaximal aerobic exercise in healthy adults and in GH-deficient adults. DESIGN: Eight healthy males (mean+/-s.e. age, 40.8+/-2.9 years) and eight hypopituitary males with verified GH deficiency (mean+/-s.e. age, 40.8+/-4.7 years) underwent a baseline test of their peak aerobic capacity (VO(2) peak) and lactate threshold (LT) on a cycle ergometer, as well as an evaluation of body composition. The patients were then studied on two occasions in random order when they exercised for 45 min at their LT. On one occasion, GH replacement had been discontinued from the evening before, whereas on the other occasion they received their evening GH in addition to an intravenous infusion of GH (0.4 IU) during exercise the following day. The healthy subjects exercised at their LT on one occasion without GH. RESULTS: The patients were significantly more obese and had lower VO(2) max (corrected for body weight) and LT as compared with the control subjects. Exercise induced a peak in serum GH concentrations after 45 min in the control group (11.43+/-3.61 microg/l). Infusion of GH in the patients resulted in a peak level after 45 min, whereas no increase was detected when exercising without GH (9.77+/-2.40 (GH) vs 0.11+/-0.07 microg/l (no GH)). Plasma ghrelin levels did not change significantly with time in either study, and no correlations were detected between ghrelin levels and parameters such as GH and IGF-I levels, age or body composition. Plasma ghrelin levels were significantly lower during the study period with GH as compared with the study with no GH. CONCLUSIONS: Submaximal aerobic exercise of an intensity sufficient to stimulate GH release was not associated with significant alterations in plasma ghrelin concentrations, which indicated that systemic ghrelin is not involved in the exercise-induced stimulation of GH secretion. The observation that ghrelin levels were lower during GH replacement suggests that GH may feedback-inhibit systemic ghrelin release. PMID- 12088922 TI - Acute hyperinsulinemia is associated with increased circulating levels of adrenomedullin in patients with type 2 diabetes mellitus. AB - OBJECTIVE: To investigate the effect of acute hyperinsulinemia on the plasma levels of adrenomedullin (AM) in patients with type 2 diabetes mellitus. DESIGN: We measured the plasma levels of AM in 18 patients with type 2 diabetes mellitus and in 19 normal subjects before and during a euglycemic hyperinsulinemic clamp study (the goal was for blood sugar levels of 5.24 mmol/l and insulin levels of 1200 pmol/l). Both plasma AM and serum insulin were measured by immunoradiometric assays. RESULTS: Before the glucose clamp study there was no significant difference in the plasma levels of AM between patients with type 2 diabetes mellitus and normal subjects. During the glucose clamp study, the serum levels of insulin significantly increased (from 33.0+/-3.6 to 1344.6+/-67.8 pmol/ml, P<0.001), as did the plasma levels of AM (from 12.8+/-0.7 to 14.2+/-0.9 fmol/ml, P<0.03) only in patients with type 2 diabetes mellitus. There was a significant correlation between the change in circulating levels of insulin and AM (r=0.755, P<0.01). CONCLUSIONS: Acute hyperinsulinemia induced a significant increase in the plasma levels of AM in patients with type 2 diabetes mellitus. Increased insulin may regulate circulating levels of AM in patients with type 2 diabetes mellitus. PMID- 12088923 TI - Dopaminergic tone and obesity: an insight from prolactinomas treated with bromocriptine. AB - OBJECTIVE: It has recently been shown that increased body weight is associated with prolactinomas and that weight loss occurs with normalization of prolactin levels. On the other hand, decreased dopaminergic tone in humans is well correlated with obesity. The objective of this study was to correlate changes in prolactin levels with leptin and body mass index (BMI) in patients with prolactinomas treated with the long-acting dopamine agonist bromocriptine (BC). METHODS: Eleven female and twelve male patients, aged 36.7+/-2.6 years with BMI in males of 30.4+/-1.7 kg/m(2) and in females of 24.4+/-1.2 kg/m(2), were evaluated after 1 and 6 months and 11 patients were further evaluated after 2 years of BC therapy. Plasma prolactin is presented as the mean of four samples taken daily. Serum leptin was determined in the pooled serum from three samples taken at 15-min intervals at 0800 h after an overnight fast. Multivariate linear regression and repeated measures analysis of covariance were used. RESULTS: In males, pretreatment prolactin levels were 71 362+/-29 912 mU/l while leptin levels were 14.9+/-1.8 microg/l. In females, pretreatment prolactin levels were 11 395+/-5839 mU/l and leptin levels were 16.7+/-2.5 microg/l. The sexual dimorphism of serum leptin levels at initial presentation was preserved after adjusting for BMI and prolactin-induced hypogonadism. After 1 month of therapy, prolactin levels significantly decreased (males: 17 618+/-8736 mU/l, females: 3686+/-2231; P<0.05), BMI did not change (males: 30.2+/-1.7 kg/m(2), females: 24.1+/-1.2 kg/m(2); P>0.05), while serum leptin levels decreased (males: 12.5+/ 1.5 microg/l, females: 13.6+/-2.1 microg/l; P<0.05). After 6 months of treatment, prolactin further decreased (males: 3456+/-2101 mU/l, females: 677+/-360 mU/l; P<0.05) as did BMI (males: 28.6+/-1.6 kg/m(2), females 23.1+/-1.0 kg/m(2); P<0.05). The difference was more pronounced in male patients. Leptin levels were 12.8+/-2.8 microg/l in males and 12.9+/-1.8 microg/l in females (P<0.05). After 2 years of BC treatment, prolactin levels were near normal (males: 665+/-439 mU/l, females 447+/-130 mU/l; P<0.05) and BMI remained 26.5+/-1.9 kg/m(2) for males and 23.6+/-0.8 kg/m(2) for females (P<0.05). Leptin levels were 9.5+/-2.2 microg/l in males and 18.7+/-3.1 microg/l in females (P<0.05). There was a gradual increase in the gender difference in serum leptin levels over time. Changes in serum leptin levels significantly correlated with changes in BMI (r=0.844, P<0.001) but did not correlate with changes in plasma prolactin levels after 1 month (r=0.166), 6 months (r=0.313) and 2 years (r=0.234, P>0.05). CONCLUSION: The long acting dopamine agonist BC, by increasing dopaminergic tone, may influence body weight and likely body composition by mechanisms in addition to reducing hyperprolactinemia in patients with prolactinomas. PMID- 12088924 TI - The value of plasma markers for the clinical behaviour of phaeochromocytomas. AB - OBJECTIVE: Phaeochromocytomas (PCCs) are widely known for their clinical unpredictability. This study intends to define predictive plasma markers for their variable postoperative behaviour. Furthermore, the diagnostic accuracy of these plasma tests was determined. DESIGN AND METHODS: A retrospective correlative study was performed in a series of 83 operated and four autopsied patients in order to correlate preoperative catecholamine (CAT) levels of 103 PCCs with their clinical behaviour. In a subset of cases, chromogranin-A (Chr-A) and enzymes/precursors of the CAT biosynthesis were studied for their predictive value. RESULTS: Basal CAT levels were elevated in 81/87 instances (sensitivity: 93%). Four of six cases with normal measurements showed only medullary hyperplasia. Larger PCCs, particularly those showing necrosis, capsular and vascular invasion, secreted higher CAT levels. Bilateral, hereditary tumours were less productive than their unilateral counterparts. Extra-adrenal PCCs secreted significantly lower levels of epinephrine (EPI) than intra-adrenal tumours. Fourteen patients developed metastases. According to Kaplan-Meier estimations, patients with higher levels of dopamine, norepinephrine (NE) and aromatic l-amino acid decarboxylase as well as lower ratios of EPI/EPI+NE, had significantly shorter metastases-free intervals. Existence of preoperative hypertension, left ventricular hypertrophy and measured blood pressures showed significant positive relationships with CAT levels, but not with Chr-A. CONCLUSIONS: These data showed that plasma CAT measurement is a sensitive method in the diagnostic work-up of PCCs. Those tumours producing normal levels are commonly small and asymptomatic. Furthermore, certain secretion patterns are indicative of the presence of metastases as well as the size and site of sporadic and syndrome-related PCCs. PMID- 12088926 TI - Evidence that luteinising hormone receptor polymorphisms may contribute to male undermasculinisation. AB - BACKGROUND: The luteinising hormone receptor (LHR) is necessary for the stimulation of androgen production and male genital development. It contains three protein polymorphisms: a leucine and glutamine insertion between codons 8 and 9 (LQ+) and two amino acid substitutions (N291S, N312S). OBJECTIVES: To determine whether these LHR polymorphisms are associated with male genital undermasculinisation or the androgen receptor polyglutamine repeat polymorphism (AR(Q)n), which contributes in some cases to the cause of genital undermasculinisation. METHODS: The LHR polymorphisms were assessed by PCR amplification of genomic DNA, followed by restriction enzyme analysis. The frequency of the LHR polymorphisms were compared between an undermasculinised male group (n=75) and a control group (n=55). RESULTS: LQ+ was not independently associated with the undermasculinised group (P=0.09), but it was associated with increased AR(Q)n within the undermasculinised group (P=0.02), particularly for AR(Q)n lengths >or=26 (P=0.002). In the undermasculinised group, homozygosity for N291 (872A/A) was more frequent (P=0.05), whereas homozygosity for N312 (935A/A) was less frequent (P=0.05). The combination of the presence of 872A/A and the absence of 935A/A showed a stronger association with the undermasculinised group than either polymorphism independently (P=0.006). The odds ratio of this genotype compared with any other, between the undermasculinised and control groups was 3.28 (95% confidence interval (CI) 1.33 to 8.08). CONCLUSION: LHR polymorphisms may contribute to genital undermasculinisation. PMID- 12088925 TI - Serum inhibin B, FSH, LH and testosterone levels before and after human chorionic gonadotropin stimulation in prepubertal boys with cryptorchidism. AB - BACKGROUND: Several studies have indicated that cryptorchidism is associated with degenerative changes in both Sertoli cells and germ cells. The gonadal peptide hormone inhibin B reflects Sertoli cell function. Low inhibin B levels are found in a large portion of formerly cryptorchid men who show compromised seminiferous tubule function. It is not known if inhibin B can be used to demonstrate early damage of seminiferous tubules in prepubertal boys with cryptorchidism. METHODS: We investigated the relationship between serum levels of inhibin B, testosterone, FSH and LH in 62 prepubertal boys with uni- and bilateral cryptorchidism. Furthermore, we investigated the changes in serum levels of inhibin B and the corresponding changes in serum levels of FSH, LH and testosterone during a short course (3 weeks) of human chorionic gonadotropin (hCG) injections in 18 of these cryptorchid boys. RESULTS: In the 62 prepubertal boys with uni- or bilateral cryptorchidism there were no significant differences in baseline levels (median and range) of inhibin B (88 (20-195) pg/ml vs 78 (35-182) pg/ml; not significant), LH (0.08 (<0.05-0.99) IU/l vs 0.06 (<0.05-1.61) IU/l; not significant) and FSH (0.60 (0.08-3.73) IU/l vs 0.85 (0.25-2.55); not significant) compared with 156 healthy prepubertal boys, and there were no differences in hormonal levels between boys with uni- or bilateral cryptorchidism. There was no correlation between baseline levels of inhibin B and FSH. In boys younger than 9 years, we found no correlation between baseline levels of inhibin B and LH whereas, in boys older than 9 years, baseline levels of inhibin B were positively correlated to baseline LH (Spearman rank correlation coefficient ((R(s))=0.58, P=0.03). Treatment with hCG (1500 IU intramuscularly twice weekly for 3 weeks) resulted in descensus of testes in 9 out of 18 patients. In all boys but one, irrespective of age, hCG induced a marked increase in testosterone into the adult range (from undetectable to 21.8 (7.0-35.4) nmol/l; P<0.001) and completely suppressed FSH and LH levels. Serum levels of inhibin B increased significantly from 116 (50-195) pg/ml to 147 (94-248) pg/ml (P<0.05), but not uniformly. The increase in serum levels of inhibin B was inversely correlated to baseline inhibin B (Rs=-0.52, P=0.03) and baseline FSH (R(s)=-0.59, P<0.01). CONCLUSIONS: We therefore suggest that, in the prepubertal testes, inhibin B is secreted from the prepubertal Sertoli cells following hCG, whereas early pubertal testes with more differentiated Sertoli cells are not able to secrete inhibin B in response to hCG stimulation, perhaps due to lack of germ cell-derived betaB-subunits. We found (a) normal inhibin B levels in prepubertal boys with uni- or bilateral cryptorchidism, (b) that hCG stimulated testosterone markedly and suppressed FSH and LH levels and (c) that hCG treatment stimulated inhibin B levels in the youngest cryptorchid boys. In the oldest prepubertal boys no hCG-induced changes in inhibin B were shown. PMID- 12088927 TI - Expression of G(alpha)(s) proteins and TSH receptor signalling in hyperfunctioning thyroid nodules with TSH receptor mutations. AB - OBJECTIVE: Constitutively activating mutations of the thyrotrophin receptor (TSHR) are the main molecular cause of hyperfunctioning thyroid nodules (HTNs). The G protein coupling is an important and critical step in the TSHR signalling which mainly includes G(alpha)(s), G(alpha)(i) and G(alpha)(q)/11 proteins. DESIGN: We investigated the in vitro consequences of overexpressing G(alpha) proteins on signalling of the wild-type (WT) or mutated TSHR. Moreover, we investigated whether changes in G(alpha) protein expression are pathophysiologically relevant in HTNs or cold thyroid nodules (CTNs). METHODS: Wild-type TSH receptor and mutated TSH receptors were coexpressed with G(alpha)(s), G(alpha)(i) or G(alpha)(q)/11, and cAMP and inositol phosphate (IP) production was measured after stimulation with TSH. The expression of G(alpha)(s), G(alpha)(i) and G(alpha)(q)/11 proteins was examined by Western blotting in 28 HTNs and 14 CTNs. RESULTS: Coexpression of G(alpha)(s) with the WT TSH receptor in COS 7 cells significantly increased the basal and TSH-stimulated cAMP accumulation while coexpression of the G(alpha)(q) or G(alpha)11 protein significantly increased the production of cAMP and inositol triphosphate (IP(3)). The coexpression of the TSH receptor mutants (I486F, DEL613-621), known to couple constitutively to G(alpha)(s) and G(alpha)(q) with G(alpha)(s) and G(alpha)(q)/11, significantly increased the basal and stimulated cAMP and IP(3) accumulation. Coexpression of the TSH receptor mutant V556F with G(alpha)(s) only increased the basal and stimulated cAMP production while its coexpression with G(alpha)(q)/11 increased the basal and stimulated IP(3) signalling. The expression of G(alpha)(s) protein subunits determined by Western blotting was significantly decreased in 14 HTNs with a constitutively activating TSH receptor mutation in comparison with the corresponding surrounding tissue, while in 14 HTNs without TSH receptor or G(alpha)(s) protein mutation and in 14 CTNs the expression of G(alpha)(s) protein was not different compared with the surrounding tissue. The expression of G(alpha)(i) and G(alpha)(q)/11 proteins in HTNs or CTNs was not significantly different compared with the surrounding tissue. CONCLUSIONS: The reduced expression of G(alpha)(s) protein subunits in HTNs with TSHR mutations could act as a feedback mechanism to desensitise the chronically stimulated cAMP cascade. As G(alpha) protein expression was not significantly increased in the majority of CTNs and HTNs an influence of G(alpha) overexpression on TSH signalling could be excluded in these nodules. PMID- 12088929 TI - Origin of parathyroid hormone (PTH) fragments detected by intact-PTH assays. AB - BACKGROUND: Intact parathyroid hormone (I-PTH) assays react with non-(1-84)PTH, large carboxyl-terminal (C) fragments with a partially preserved amino-terminal (N) structure. They account for up to 50% of I-PTH in renal failure and may be implicated in PTH resistance. We wanted to know if they were secreted by the parathyroid glands and generated by peripheral metabolism of PTH(1-84). METHODS: Anesthetized normal and nephrectomized (NPX) rats were injected i.v. with 1.5 microg human (h) PTH(1-84). Blood was obtained from 8 rats at 2, 4, 6, 8, 12, 24, 48 and 96 min. I-PTH (Allegro I-PTH) was measured in all samples. Pools of serum were fractionated by HPLC at each time point and the fractions assayed to quantitate hPTH(1-84) and non-(1-84)PTH. Secretion studies were performed with dispersed cells from 5 parathyroid adenomas. The serum of 10 patients with primary hyperparathyroidism and cell supernatants were fractionated by HPLC and were analyzed as described. RESULTS: hPTH(1-84) disappeared from serum biexponentially. The half-life of the first exponential was similar in normal (2.08 min) and NPX (1.94 min) rats, while that of the second was longer in NPX rats (32.4 vs 20.9 min). The residual quantity of hPTH(1-84) under the curve was greater in NPX (6964+/-2392 pmol) than in normal rats (3229+/-561 pmol; P<0.001). Non-(1-84)PTH concentration was maximal at 8 min in both groups and was higher in NPX (92.8+/-13.8 pmol/l) than in normal rats (38.8+/-7.2 pmol/l; P<0.01). The area under the curve of non-(1-84)PTH was also greater in NPX (1904+/-405 pmol) than in normal rats (664+/-168 pmol; P<0.001). All parathyroid adenomas secreted non-(1-84)PTH. It represented 21.1+/-3.9% of secreted and 32.5+/-1.3% of circulating I-PTH in primary hyperparathyroidism. CONCLUSIONS: Non-(1-84)PTH, like other C-PTH fragments, originates from both the peripheral metabolism of hPTH(1-84) and from parathyroid gland secretion. Renal failure influences its concentration by increasing the amount of substrate available and by reducing non (1-84)PTH clearance. Its higher proportion in serum relative to cell supernatants in primary hyperparathyroidism reflects the added role of peripheral metabolism and the longer half-life of fragments. PMID- 12088928 TI - Nitric oxide synthase activity in hyperthyroid and hypothyroid rats. AB - OBJECTIVE: Thyroid disorders are accompanied by important changes in haemodynamic and cardiac functions and renal sodium handling. Since nitric oxide (NO) plays a crucial role in regulating vascular tone and renal sodium excretion, the present paper was designed to determine whether changes in the activity of NO synthase (NOS) participate in the cardiovascular and renal manifestations of thyroid disorders. METHODS: We measured NOS activity in the heart (left and right ventricles), vessels (aorta and cava) and kidney (cortex and medulla) of euthyroid, hyperthyroid and hypothyroid rats after 6 weeks of treatment. NOS activity was determined by measuring the conversion of L-[(3)H]-arginine to L [(3)H]-citruline. RESULTS: NOS activity was higher in all tissues from hyperthyroid rats when compared with controls, except in the right ventricle. In the hypothyroid group, NOS activity showed a more heterogeneous pattern, with significant increases in both ventricles but significant reduction in the aorta, while in the vena cava, renal cortex and medulla the enzyme activity also tended to be higher, but significance was not reached. CONCLUSIONS: These data indicated that NOS activity was upregulated in tissues primarily related to blood pressure control in hyperthyroid rats, suggesting that an increased NO production may contribute to the hyperdynamic circulation in hyperthyroidism and may have a protective homeostatic effect in the target organs of the hypertension that accompanies this endocrine disease. The aortic and renal findings in hypothyroid rats suggested a possible role for NOS in the increased peripheral resistance and the normal pressure-diuresis-natriuresis response of these hypotensive animals, although hypothyroidism produced a heterogeneous tissue response in NOS activity. PMID- 12088930 TI - Nateglinide, but not repaglinide, stimulates growth hormone release in rat pituitary cells by inhibition of K channels and stimulation of cyclic AMP dependent exocytosis. AB - OBJECTIVE: GH causes insulin resistance, impairs glycemic control and increases the risk of vascular diabetic complications. Sulphonylureas stimulate GH secretion and this study was undertaken to investigate the possible stimulatory effect of repaglinide and nateglinide, two novel oral glucose regulators, on critical steps of the stimulus-secretion coupling in single rat somatotrophs. METHODS: Patch-clamp techniques were used to record whole-cell ATP-sensitive K(+) (K(ATP)) and delayed outward K(+) currents, membrane potential and Ca(2+) dependent exocytosis. GH release was measured from perifused rat somatotrophs. RESULTS: Both nateglinide and repaglinide dose-dependently suppressed K(ATP) channel activity with half-maximal inhibition being observed at 413 nM and 13 nM respectively. Both compounds induced action potential firing in the somatotrophs irrespective of whether GH-releasing hormone was present or not. The stimulation of electrical activity by nateglinide, but not repaglinide, was associated with an increased mean duration of the action potentials. The latter effect correlated with a reduction of the delayed outward K(+) current, which accounts for action potential repolarization. The latter effect had a K(d) of 19 microM but was limited to 38% inhibition. When applied at concentrations similar to those required to block K(ATP) channels, nateglinide in addition potentiated Ca(2+) evoked exocytosis 3.3-fold (K(d)=3 microM) and stimulated GH release 4.5-fold. The latter effect was not shared by repaglinide. The stimulation of exocytosis by nateglinide was mimicked by cAMP and antagonized by the protein kinase A inhibitor Rp-cAMPS. CONCLUSION: Nateglinide stimulates GH release by inhibition of plasma membrane K(+) channels, elevation of cytoplasmic cAMP levels and stimulation of Ca(2+)-dependent exocytosis. By contrast, the effect of repaglinide was confined to inhibition of the K(ATP) channels. PMID- 12088931 TI - Effect of the GH secretagogue L-163,255 and restricted feeding time on GH pulsatility in the rat. AB - OBJECTIVE: To determine the effect of repeated treatments with the growth hormone secretagogue (GHS) L-163,255 on the pulsatile release of GH when administered in meal-fed rats before and after feeding. DESIGN: The first group of rats (AL, n=6) had food available ad libitum. The second (restricted, R, n=6), third (GHSB, n=6), and fourth (GHSA, n=6) groups were fed from 1100 to 1400 h. Groups GHSB and GHSA were given GHS by gavage, 3.0 mg/kg L-163,255, at 1000 h (before feeding, B) and at 1500 h (after feeding, A) respectively. Three weeks after the initiation of the treatment, blood samples were collected at 10-min intervals over 6 h, and GH levels were determined. RESULTS: In group R, the concentrations of GH were higher before feeding (17.6+/-2.4 ng/ml) than during feeding (11.2+/-1.2 ng/ml), P<0.05. The average concentrations of the peak in response to GHS were higher when GHS was administered before (121.70+/-33.68 ng/ml) than after (49.67+/-17.87 ng/ml) feeding. The mass of GH, as calculated by deconvolution analysis was also higher in the GHSB group than in the GHSA group (251.6+/-64.1 ng/ml per min vs 85.3+/-22.9 ng/ml per min respectively, P<0.05). CONCLUSION: L-163,255 is effective in inducing GH release after repeated oral administration in rats. The effectiveness is greater when GHS is administered before rather than after feeding in meal-fed animals. PMID- 12088932 TI - Endurance training improves responsiveness to insulin and modulates insulin signal transduction through the phosphatidylinositol 3-kinase/Akt-1 pathway. AB - BACKGROUND: Endurance training increases insulin-stimulated muscle glucose transport and leads to improved metabolic control in diabetic patients. OBJECTIVE: To analyze the effects of endurance training on the early steps of insulin action in muscle of rats. DESIGN: Male rats submitted to daily swimming for 6 weeks were compared with sedentary controls. At the end of the training period, anesthetized animals received an intravenous (i.v.) injection of insulin and had a fragment of their gastrocnemius muscle excised for the experiments. METHODS: Associations between insulin receptor, insulin receptor substrates (IRS) 1 and -2 and phosphatidylinositol 3-kinase (PI3-kinase) were analyzed by immunoprecipitation and immunoblotting. Akt-1 serine phosphorylation and specific protein quantification were detected by immunoblotting of total extracts, and IRS 1/IRS-2-associated PI3-kinase activity were determined by thin-layer chromatography. RESULTS: Insulin-induced phosphorylation of IRS-1 and IRS-2 increased respectively by 1.8-fold (P<0.05) and 1.5-fold (P<0.05), whereas their association with PI3-kinase increased by 2.3-fold (P<0.05) and 1.9-fold (P<0.05) in trained rats as compared with sedentary controls, respectively. The activity of PI3-kinase associated with IRS-1 and IRS-2 increased by 1.8-fold (P<0.05) and 1.7-fold (P<0.05) respectively, in trained rats as compared with their untrained counterparts. Serine phosphorylation of Akt-1/PKB increased 1.7-fold (P<0.05) in trained rats in response to insulin. These findings were accompanied by increased responsiveness to insulin as demonstrated by a reduced area under the curve for insulin during an i.v. glucose tolerance test, by increased glucose disappearance rate during an insulin tolerance test, and by increased expression of glucose transporter-4. CONCLUSIONS: The increased responsiveness to insulin induced by chronic exercise in rat skeletal muscle may result, at least in part, from the modulation of the insulin signaling pathway at different molecular levels. PMID- 12088933 TI - Influence of thyroid status and growth hormone deficiency on ghrelin. AB - OBJECTIVE: To assess whether some of the alterations in energy homeostasis present in thyroid function disorders and GH deficiency could be mediated by ghrelin. DESIGN: To assess the influence of thyroid status on ghrelin, adult male Sprague-Dawley rats were treated with vehicle (euthyroid), amino-triazole (hypothyroid) or l-thyroxine (hyperthyroid). The influence of GH on ghrelin was assessed in wild-type (control) and GH-deficient (dwarf) Lewis rats. Evaluation of gastric ghrelin mRNA expression in the stomach was carried out by Northern blot. Circulating levels of ghrelin were measured by radioimmunoassay. RESULTS: Hypothyroidism resulted in an increase in gastric ghrelin mRNA levels (euthyroid: 100+/-3.2% vs hypothyroid: 127.3+/-6.5%; P<0.01), being decreased in hyperthyroid rats (70+/-5.4%; P<0.01). In keeping with these results, circulating plasma ghrelin levels were increased in hypothyroid (euthyroid: 124+/-11 pg/ml vs hypothyroid: 262+/-39 pg/ml; P<0.01) and decreased in hyperthyroid rats (75+/-6 pg/ml; P<0.01). Using an experimental model of GH deficiency, namely the dwarf rat, we found a decrease in gastric ghrelin mRNA levels (controls: 100+/-6% vs dwarf: 66+/-5.5%; P<0.01) and circulating plasma ghrelin levels (controls: 124+/ 12 pg/ml vs dwarf: 81+/-7 pg/ml; P<0.01). CONCLUSION: This study provides the first evidence that ghrelin gene expression is influenced by thyroid hormones and GH status and provides further evidence that ghrelin may play an important role in the alteration of energy homeostasis and body weight present in these pathophysiological states. PMID- 12088934 TI - Serum creatinine patterns in coronary bypass surgery patients with and without postoperative cognitive dysfunction. AB - Renal dysfunction is common after coronary artery bypass graft (CABG) surgery. We have previously shown that CABG procedures complicated by stroke have a threefold greater peak serum creatinine level relative to uncomplicated surgery. However, postoperative creatinine patterns for procedures complicated by cognitive dysfunction are unknown. Therefore, we tested the hypothesis that postoperative cognitive dysfunction is associated with acute perioperative renal injury after CABG surgery. Data were prospectively gathered for 282 elective CABG surgery patients. Psychometric tests were performed at baseline and 6 wk after surgery. Cognitive dysfunction was defined both as a dichotomous variable (cognitive deficit [CD]) and as a continuous variable (cognitive index). Forty percent of patients had CD at 6 wk. However, the association between peak percentage change in postoperative creatinine and CD (parameter estimate = -0.41; P = 0.91) or cognitive index (parameter estimate = -1.29; P = 0.46) was not significant. These data indicate that postcardiac surgery cognitive dysfunction, unlike stroke, is not associated with major increases in postoperative renal dysfunction. IMPLICATIONS: We previously noted that patients with postcardiac surgery stroke also have greater acute renal injury than unaffected patients. However, in the same setting, we found no difference in renal injury between patients with and without cognitive dysfunction. Factors responsible for subtle postoperative cognitive dysfunction do not appear to be associated with clinically important renal effects. PMID- 12088935 TI - Myocardial ischemia and cytokine response are associated with subsequent onset of infections after noncardiac surgery. AB - Postoperative myocardial ischemia (POMI) is prevalent among patients after major noncardiac surgery. Surgery, as well as POMI, may modulate the immune system, potentially worsening patient outcome. We sought to investigate the modulation of soluble interleukin (IL)-6 and IL-10 by POMI and its association with increased postoperative infection rates. Two-hundred-three patients undergoing elective major abdominal, vascular, and orthopedic surgery participated in this prospective observational study. Perioperative management was standardized. Hemodynamic variables were kept within 20% of baseline. POMI was assessed by Holter electrocardiography starting at least 8 h before the induction of anesthesia and continued until 96 h after surgery. Twelve-lead electrocardiograms, cardiac enzymes, and immune variables were obtained at the time of admission to the hospital, before surgery, before the induction of anesthesia, after surgery, at the time of admission to the intensive care unit, and 6, 12, 18, 24, 36, 48, 72, 96, 120, 144, and 168 h after surgery. Infections were diagnosed according to the Centers for Disease Control criteria. The incidence of POMI was 27%, and the majority of cases (76%) occurred within the first 24 h after surgery. IL-6 and IL-10 levels significantly increased during surgery but did not differ between the POMI and Non-POMI groups. However, in the subset of patients who developed severe infections or sepsis (n = 47) a median of 3 days (range, 1-8 days) after surgery, the intraoperative increases of IL-6 and IL-10 in the POMI group were, respectively, 3 and 10 times higher compared with the increase in the Non-POMI group. By using a multifactorial analysis in these patients with severe infections, the type of surgical trauma was associated with an increased IL-6 response, whereas the increase in IL-10 was attributed to POMI. These findings suggest that immediate cytokine responses due to POMI and type of surgery might be relevant for the later onset of severe infections and sepsis. IMPLICATIONS: Postoperative myocardial ischemia (POMI) occurred in 27% of patients after major noncardiac surgery. This was associated with an immediate augmented cytokine response in the first 12 h after surgery in patients who developed severe infections or sepsis 3 days later. POMI was associated with an increased interleukin (IL)-10 response, whereas IL-6 was associated with the type of surgery. PMID- 12088936 TI - Does dobutamine improve ventricular function in dogs with regional myocardial dysfunction? AB - We studied the effect of systemic dobutamine infusion (4 microg. kg(-1). min(-1) IV) on regional wall motion abnormalities (RWMAs) in eight anesthetized open chested dogs. We hypothesized that infusion of small doses of dobutamine would reduce RWMAs and improve global ventricular function. Apical RWMAs were induced by local intracoronary boluses of 9.0 mg esmolol. Phase angles, effective stroke volume (SV), maximum SV, stroke work, and segmental shortening were compared among four left ventricular (LV) regions (apical, papillary, chordal, and basal) during baseline, dobutamine, esmolol, and dobutamine-esmolol treatments. The minimal global LV volume was designated as 0 degrees, and the cardiac cycle was divided into 360 intervals. Regional phase angles were defined as the distance (in degrees) that regional minimum volume differed from global minimal LV volume (end-systole). RWMA decreased blood pressure (92 +/- 2 mm Hg to 84 +/- 3 mm Hg) and increased LV end-diastolic pressure (1.8 +/- 0.5 mm Hg to 4.2 +/- 0.8 mm Hg). RWMA delayed regional contraction (-2.9 degrees +/- 1.6 degrees to 52.3 degrees +/- 1.5 degrees ) and decreased effective SV (2.3 +/- 0.4 mL to 1.6 +/- 0.3 mL) in the affected apical region but did not decrease maximal SV. Systemic infusion of dobutamine restored global LV function but failed to eliminate RWMA, as evidenced by decreased apical synchrony, effective SV, and stroke work. We concluded that systemic dobutamine restored global LV function but failed to correct RWMA. IMPLICATIONS: We examined the effect of systemic dobutamine on regional wall motion abnormalities (RWMAs) induced by intracoronary esmolol infusion in eight anesthetized dogs. Esmolol dilated the heart and decreased regional synchrony of contraction. Dobutamine restored cardiac function but failed to correct the asynchrony of regional contraction caused by esmolol induced RWMAs. PMID- 12088937 TI - The plasma supplemented modified activated clotting time for monitoring of heparinization during cardiopulmonary bypass: a pilot investigation. AB - The standard celite or kaolin activated clotting time (ACT) correlates poorly with heparin levels during cardiopulmonary bypass (CPB). We compared a modified kaolin ACT, in which plasma was supplemented, to a standard undiluted kaolin ACT for monitoring heparin levels during CPB. Fifteen patients undergoing normothermic CPB were enrolled in this prospective study. Heparin management was performed according to the Hepcon HMS results (Medtronic, Minneapolis, MN). The ACTs were performed with the ACT II device (Medtronic). Hepcon HMS calculations, standard kaolin ACTs, and plasma supplemented modified ACTs (mACTs), prepared by diluting blood samples 1:1 with human plasma (Behring, Marburg, Germany), were measured every 30 min during CPB. The data obtained were correlated to the plasma chromogenic anti-Xa activity as a reference assay for heparin levels. A total of 64 samples were evaluated. The chromogenic anti-Xa activity ranged from 0.2 to 5.5 IU/mL. The Hepcon HMS calculations ranged from 2.7-8.2 IU/mL of heparin, the standard ACT ranged from 424 to >999 s, and the mACT ranged from 210 to 801 s. The correlation to the chromogenic anti-Xa method was r = 0.43 for the standard kaolin ACT and r = 0.69 for the plasma mACT. The plasma mACT provided an improved correlation to chromogenically measured levels of anti-Xa activity during CPB. The improved correlation most likely results from a correction of the effects of the impairment of the coagulation system caused by hemodilution and consumption of procoagulants on extracorporeal surfaces. IMPLICATIONS: During cardiopulmonary bypass, the plasma modified kaolin activated clotting time (ACT) provides a better correlation with heparin levels than the standard kaolin ACT. PMID- 12088938 TI - The paradoxical positive inotropic effect of sevoflurane in healthy and cardiomyopathic hamsters. AB - We investigated the effects of sevoflurane (0.7 to 3.6 vol%) on inotropy and lusitropy in left ventricular papillary muscles of healthy hamsters and genetically induced cardiomyopathic (strain BIO 14.6) hamsters in vitro (29 degrees C, pH 7.40, Ca(2+) 2.5 mM, stimulation frequency three per minute) under low (isotony) and high (isometry) loads. Sevoflurane induced a moderate positive inotropic effect in healthy hamsters (maximum unloaded shortening velocity and isometric active force at 3.6 vol%: 115% +/- 12% and 128% +/- 21% of baseline values, respectively; P < 0.01) and in cardiomyopathic hamsters (maximum unloaded shortening velocity and isometric active force at 3.6 vol%: 115% +/- 20% and 124% +/- 31% of baseline values, respectively; P < 0.05). This positive inotropic effect did not differ between healthy and cardiomyopathic hamsters, even when sevoflurane concentrations were corrected for minimum alveolar anesthetic concentration values in each strain, and was unchanged after alpha- and beta adrenoceptor blockade. After calcium-channel blockade, this positive inotropic effect was abolished in healthy hamsters but enhanced in cardiomyopathic hamsters. In both strains, sevoflurane induced a moderate negative lusitropic effect under low and high loads. IMPLICATIONS: A paradoxical moderate positive inotropic effect of sevoflurane was observed in hamster ventricular muscle. This effect was likely related to calcium channel interaction, because after calcium channel blockade, it was abolished in healthy hamsters and enhanced in cardiomyopathic hamsters. PMID- 12088939 TI - Life-threatening hyperkalemia: a complication of spironolactone for heart failure in a patient with renal insufficiency. AB - IMPLICATIONS: Serum potassium concentration should be measured immediately before operation to detect hyperkalemia in heart failure patients treated with spironolactone. Renal insufficiency, advanced age, potassium supplementation, decompensated congestive heart failure, and a spironolactone dose larger than 25 mg/d increase the risk of hyperkalemia as a consequence of spironolactone therapy. PMID- 12088940 TI - The anesthetic considerations in patients with ventricular assist devices presenting for noncardiac surgery: a review of eight cases. AB - IMPLICATIONS: The number of patients supported by ventricular assist devices (VADs) that present for noncardiac surgery is increasing in our institution. Our recent experience with eight such patients is reported, along with a review of the most commonly implanted VADs and the anesthetic implications and considerations for VAD-supported patients undergoing noncardiac surgery. PMID- 12088941 TI - Shy-Drager syndrome and severe unexplained intraoperative hypotension responsive to vasopressin. AB - IMPLICATIONS: We describe the first case of Shy-Drager syndrome diagnosed on the basis of intraoperative hemodynamic changes. The initial hypertension in the supine position followed by severe hypotension after hydralazine administration, ultimately responsive to vasopressin, led to a diagnosis of Shy-Drager syndrome. We suggest that vasopressin may be the drug of choice in patients with Shy-Drager syndrome with refractory hypotension. PMID- 12088942 TI - Profound sinus bradycardia after intravenous nicardipine. AB - IMPLICATIONS: Nicardipine-induced bradycardia has been reported in experimental animals but not in clinical patients. We report a clinical case of unexpected bradycardia caused by nicardipine. The mechanism of this bradycardia was not clear, and depression of sympathetic tone by epidural anesthesia, hypothermia, and paroxysmal atrial fibrillation might have been contributory. PMID- 12088943 TI - The effect on lung mechanics in anesthetized children with rapacuronium: a comparative study with mivacurium. AB - The administration of rapacuronium increases the risk of severe bronchospasm. There have been no studies of pulmonary function directly demonstrating airway constriction with rapacuronium in children. In this study, 10 ASA physical status I or II patients (aged 2-6 yr) were randomly divided into 2 equal groups, receiving either rapacuronium or mivacurium. Anesthesia was induced with sevoflurane and maintained with remifentanil (0.2-0.3 microg. kg(-1). min(-1)) and propofol (200-250 microg. kg(-1). min(-1)) infusions. We performed three sets of pulmonary function tests: baseline, after the administration of muscle relaxant, and after the administration of a beta(2) agonist. In both groups, there were no changes in static respiratory compliance. The increase in total respiratory system resistance after the administration of rapacuronium did not reach statistical significance (214.4% +/- 122.65% of baseline, P approximately 0.1), whereas maximal expiratory flow at 10% of forced vital capacity (MEF)(10) and MEF(functional residual capacity) on partial flow-volume curves by the forced deflation technique decreased markedly (53.4% +/- 18.49%, P < 0.01 and 41.3% +/- 27.42%, P < 0.001, respectively). With the administration of mivacurium, no changes were observed in respiratory system resistance (109.5% +/- 30.28%). MEF(10) decreased slightly (77.0% +/- 9.03%, P < 0.005) whereas MEF(FRC) did not (81.2% +/- 29.85%, not significant). After the administration of a beta(2) agonist, all measurements returned to baseline. Thus, the administration of rapacuronium consistently results in lower airway obstruction with minimal changes in static respiratory compliance when compared with mivacurium. IMPLICATIONS: Pulmonary function tests in the present study showed that rapacuronium consistently causes severe bronchoconstriction, confirming clinical case reports of bronchospasm. The bronchoconstriction is reversible with albuterol. Mivacurium also causes very mild subclinical bronchoconstriction. PMID- 12088944 TI - An evaluation of a virtual reality airway simulator. AB - In this research, we sought to test the hypothesis that the AccuTouch Flexible Bronchoscopy Simulator (Simulator) is an effective way to teach clinicians the psychomotor skills necessary to use the fiberoptic bronchoscope as an instrument for intubating the trachea of a pediatric patient. Pediatric residents with no prior experience in fiberoptic bronchoscopy were studied. Residents performed fiberoptic intubation on children undergoing general anesthesia. Tapes of these intubations were analyzed for: time to visualization of the carina, and number and time that the bronchoscope tip hit the mucosa. Residents were then trained on the Simulator. Performance of fiberoptic intubation on a subsequent child was compared. Training on the Simulator was the only instruction that the residents received between the two cases. A control group of residents performed two consecutive intubations without training on the Simulator between cases. Residents studied an average of 17 cases, and spent 39 min on the Simulator. Performance was markedly improved after the Simulator. Time to completion of successful intubation with a bronchoscope was reduced from 5.15 to 0.88 min (P < 0.001). The number of times that the tip of the bronchoscope hit the mucosa was reduced from 21.4 to 3.0 (P < 0.001). The amount of time that the resident spent viewing the mucosa decreased from 2.24 to 0.19 min (P < 0.001). The percent of time viewing the channel of the airway increased from 58.5% to 80.4% (P = 0.004). This bronchoscopy simulator was very effective in teaching residents the psychomotor skills necessary for fiberoptic intubation. Significant improvement was seen in time to completion of endotracheal intubation, as well as other performance indicators. IMPLICATIONS: This research showed that the AccuTouch Bronchoscopy Simulator is an effective way to teach the psychomotor skills necessary to intubate the trachea of patients using a fiberoptic bronchoscope. The residents that practiced on the Simulator dramatically improved their skills compared with a control group of residents. PMID- 12088945 TI - Insufficiency in a new temporal-artery thermometer for adult and pediatric patients. AB - SensorTouch is a new noninvasive temperature monitor and consists of an infrared scanner that detects the highest temperature on the skin of the forehead, presumably over the temporal artery. The device estimates core temperature (T(core)). We tested the hypothesis that the SensorTouch is sufficiently precise and accurate for routine clinical use. We studied adults (n = 15) and children (n = 16) who developed mild fever, a core temperature of at least 37.8 degrees C, after cardiopulmonary bypass. Temperature was recorded at 15-min intervals throughout recovery with the SensorTouch thermometer and from the pulmonary artery (adults) or bladder (children). Pulmonary artery (T(core)) and SensorTouch (T(st)) temperatures correlated poorly in adults: T(core) = 0.7. T(st) + 13, r(2) = 0.3. Infrared and pulmonary artery temperatures differed by 1.3 +/- 0.6 degrees C; 89% of the adult temperatures thus differed by more than 0.5 degrees C. Bladder and infrared temperatures correlated somewhat better in pediatric patients: T(core) = 0.9. T(st) + 12, r(2) = 0.6. Infrared and bladder temperatures in children differed by only 0.3 degrees C, but the SD of the difference was 0.5 degrees C. Thus, 31% of the values in the infants and children differed by more than 0.5 degrees C. IMPLICATIONS: We evaluated a noninvasive infrared forehead thermometer (SensorTouch) in adult and pediatric cardiac patients. Accuracy was poor in the adults and suboptimal in infants and children. PMID- 12088946 TI - Awareness and recall in outpatient anesthesia. AB - We studied the incidence of awareness and explicit recall during general anesthesia in outpatients versus inpatients undergoing surgery. During a 14.5-mo period, we structurally interviewed 1500 outpatients and 2343 inpatients. Among outpatients, there were five cases of awareness and recall (one with clear intraoperative recollections and four with doubtful intraoperative recollections). Of the inpatients, six reported awareness and recall (three with clear and three with doubtful intraoperative recollections). The incidence of clear intraoperative recollections was 0.07% in outpatients and 0.13% in inpatients. The difference in the incidence was not significant. Among outpatients, those with awareness and recall were given smaller doses of sevoflurane than those without awareness and recall (P < 0.05). In conclusion, awareness and recall are rare complications of general anesthesia, and outpatients are not at increased risk for this event compared with inpatients undergoing general anesthesia. IMPLICATIONS: Rapid recovery from general anesthesia is a crucial element of outpatient surgery. However, this practice may predispose a patient to receive less anesthetic, with increased risk for awareness and recall. We have shown that outpatients undergoing an operation using general anesthesia are not at increased risk for awareness compared with inpatients. PMID- 12088947 TI - Development of an appropriate list of surgical procedures of a specified maximum anesthetic complexity to be performed at a new ambulatory surgery facility. AB - A common but difficult task for a hospital when it decides to open a freestanding ambulatory surgery facility is how to decide which surgical procedures should be done at the new facility. This is necessary in order to determine how many operating rooms to plan for the new facility and which ancillary services are needed on-site. In this case study, we describe a novel methodology that we used to develop a comprehensive list of procedures to be done at a new ambulatory facility. The level of anesthetic complexity of a procedure was defined by its number of ASA Relative Value Guide basic units. Broad categories of procedures (e.g., eye surgery) were defined according to the International Classification of Diseases, Ninth Revision, Clinical Modification. We identified 22 categories that are of a type that every procedure in the category has no more than seven basic units. In addition, by analyzing all procedures that the hospital being studied actually performed on an ambulatory basis, we identified six other categories of procedures that were of a type that all procedures eligible for surgery at the new facility had seven or fewer basic units. IMPLICATIONS: We describe a novel method to develop a comprehensive list of procedures that have a prespecified maximum level of anesthetic complexity to be performed at a new ambulatory surgery facility. PMID- 12088948 TI - Sensitivity of the alpha7 nicotinic acetylcholine receptor to isoflurane may depend on receptor inactivation. AB - In previous studies, we demonstrated that nicotinic acetylcholine receptors (nAChRs) composed of the alpha7 subunit are unaffected by the co-application of isoflurane with agonists at concentrations up to 640 microM (two times the minimum alveolar anesthetic concentration). Modulation of alpha7-nAChR activity by isoflurane might have important behavioral ramifications because these receptors are expressed diffusely in the central and peripheral nervous systems and play pre- and postsynaptic roles in synaptic transmission. Here we have demonstrated that under some potentially physiologically relevant circumstances, the activation of alpha7 nAChRs may be inhibited by clinically relevant concentrations of isoflurane. We evaluated isoflurane inhibition of alpha7 nAChRs from chicks and humans expressed in Xenopus oocytes using two-electrode voltage clamp methodology. We determined the influence of time of preperfusion of isoflurane, agonist concentration, and membrane potential on inhibition by isoflurane. Both activation by a large concentration of agonist and isoflurane preperfusion increased inhibition. The half-maximal inhibitory concentration for isoflurane inhibition of chick alpha7 nAChR with isoflurane preperfusion and activation by 100 microM of acetylcholine was 938 +/- 26, and when activated by 1 mM of acetylcholine, it was 408 +/- 51 microM. The increase in inhibition with isoflurane preexposure and large agonist concentration raises the possibility that isoflurane interacts preferentially with a closed or closed-desensitized state of the channel. IMPLICATIONS: Nicotinic receptors expressed in the brain have been considered a possible target for the actions of isoflurane. We studied the effect of isoflurane on alpha7 type nicotinic receptors expressed in Xenopus oocytes. We find that when activated by large concentrations of acetylcholine, alpha7 nicotinic receptors are inhibited by isoflurane at concentrations near MAC. PMID- 12088949 TI - The inhibitory effect of local anesthetics on bradykinin-induced phospholipase D activation in rat pheochromocytoma PC12 cells. AB - Bradykinin induces activation of phospholipase D (PLD) via B(2) receptors in neuronal cells. To demonstrate molecular mechanism(s) of local anesthetics, we examined whether and how local anesthetics affect bradykinin-induced PLD activation in PC12 cells. Using [(3)H]Palmitic acid-labeled PC12 cells stimulated with bradykinin, formation of [(3)H]phosphatidylbutanol was measured as a variable of PLD activity. Bradykinin-stimulated PLD activity seemed to peak at 2 min. Procaine, lidocaine, ropivacaine, bupivacaine, and tetracaine suppressed the bradykinin-induced PLD activation. We chose tetracaine, the most potent drug among the local anesthetics tested, to examine how local anesthetics affect phospholipase C, protein tyrosine kinase, and extracellular signal-regulated kinase, which are the molecules upstream of PLD. Tetracaine at clinically relevant concentrations (1 approximately 10 x 10(-4) M) inhibited the bradykinin induced PLD activation in a dose- and time-dependent manner, but neither tetrodotoxin nor nifedipine affected the PLD activation. Tetracaine (5 x 10(-4) M) slightly potentiated brady-kinin-induced phospholipase C activation. Bradykinin-stimulated protein tyrosine-phosphorylation and extracellular signal regulated kinase activation were not affected by tetracaine. Tetracaine significantly decreased PLD activity of membrane fraction in PC12 cells. These results indicate that local anesthetics depress bradykinin-induced lipid signaling pathway(s) and may provide some clues to understanding the molecular mechanisms of these drugs for anesthesia or analgesia. IMPLICATIONS: Local anesthetics depressed the bradykinin-induced activation of phospholipase D (PLD) in PC12 cells. The effects of tetracaine, the most potent among the anesthetics tested, on the bradykinin-induced intracellular signaling molecules were examined. The bradykinin-induced PLD activation could be one of the potential intracellular signaling molecular sites of local anesthetic action. PMID- 12088950 TI - Electro-acupuncture at the Zusanli, Yanglingquan, and Kunlun points does not reduce anesthetic requirement. AB - We tested the hypothesis that electro-acupuncture at the Zusanli, Yanglingquan, and Kunlun acupuncture points on the legs decreases anesthetic requirement. Fourteen young, healthy volunteers were anesthetized with desflurane on two separate days. Needle electrodes were positioned at the three acupuncture points thought to produce a generalized sedative and analgesic effect. Needles were percutaneously placed on treatment days; on control days, they were insulated and taped near the insertion points. The electrodes were stimulated on the treatment day. Stimulation consisted of 2-Hz and 100-Hz currents alternated at 2-s intervals. When the end-tidal desflurane concentration of 5.5% was stable for 15 min, noxious electrical stimuli were administered via 25-gauge needles on both thighs (70 mA at 100 Hz for 10 s). Desflurane concentration was increased 0.5% when movement occurred and decreased 0.5% when it did not. An investigator, blinded to treatment, determined movement. These up-and-down sequences were continued until volunteers crossed from movement to no movement four times. A logistic regression determined the partial pressure of desflurane that produced a 50% likelihood of movement in response to noxious stimulation and consequently identified the minimum alveolar anesthetic concentration equivalent for desflurane. There was no significant difference in minimum alveolar anesthetic concentration equivalents between the electro-acupuncture (4.6% +/- 0.6%, mean +/ SD) and control (4.6% +/- 0.8%) days (P = 0.8). These data provided an 80% power for detecting a difference of 0.35 volume-percent between the groups. IMPLICATIONS: Electro-stimulation of three general acupuncture points on the leg did not reduce desflurane requirements. This type of acupuncture is thus unlikely to facilitate general anesthesia or decrease the need for anesthetic drugs. PMID- 12088951 TI - Supplementing desflurane-remifentanil anesthesia with small-dose ketamine reduces perioperative opioid analgesic requirements. AB - Relative large-dose intraoperative remifentanil could lead to the need for more postoperative analgesics. Intraoperative N-methyl-D-aspartate receptor antagonists, such as ketamine, decrease postoperative opioid use. We therefore tested the hypothesis that intraoperative small-dose ketamine improves postoperative analgesia after major abdominal surgery with remifentanil-based anesthesia. Fifty patients undergoing abdominal surgery under remifentanil-based anesthesia were randomly assigned to intraoperative ketamine or saline (control) supplementation. The initial ketamine dose of 0.15 mg/kg was followed by 2 microg. kg(-1). min(-1). In both groups, desflurane was kept constant at 0.5 minimum alveolar anesthetic concentration without N(2)O, and a remifentanil infusion was titrated to autonomic responses. All patients were given 0.15 mg/kg of morphine 30 min before the end of surgery. Pain scores and morphine consumption were recorded for 24 postoperative h. Less of the remifentanil was required in the Ketamine than in the Control group (P < 0.01). Pain scores were significantly larger in the Control group during the first 15 postoperative min but were subsequently similar in the two groups. The Ketamine patients required postoperative morphine later (P < 0.01) and received less morphine during the first 24 postoperative h: 46 mg (interquartile range, 34-58 mg) versus 69 mg (interquartile range, 41-87 mg, P < 0.01). No psychotomimetic symptoms were noted in either group. In conclusion, supplementing remifentanil-based anesthesia with small-dose ketamine decreases intraoperative remifentanil use and postoperative morphine consumption without increasing the incidence of side effects. Thus, intraoperative small-dose ketamine may be a useful adjuvant to intraoperative remifentanil. IMPLICATIONS: Supplementing remifentanil-based anesthesia with small-dose ketamine decreased intraoperative remifentanil use and postoperative morphine consumption. These data demonstrate that N-methyl-D-aspartate antagonists, such as ketamine, can be a useful adjuvant to intraoperative remifentanil. PMID- 12088952 TI - Sensitivity to vecuronium in seropositive and seronegative patients with myasthenia gravis. AB - Patients with myasthenia gravis (MG) are hypersensitive to nondepolarizing neuromuscular blocking drugs. Although antibodies to the acetylcholine receptor (AChR) often are observed in MG patients, 10% to 30% of patients do not show an anti-AChR antibody. Little is known about differences in sensitivity to nondepolarizing neuromuscular blocking drugs between MG patients with and without anti-AChR antibody. Hypothesizing that seronegative patients are as sensitive to vecuronium as seropositive patients, we assessed sensitivity in seropositive and seronegative MG patients and in non-MG patients (n = 8 each). During anesthesia with sevoflurane (2.5%) and nitrous oxide (60%) in oxygen, neuromuscular transmission was monitored by measuring the twitch tension of the adductor pollicis muscle with supramaximal stimulation. After baseline measurements, 10 microg/kg IV dose increments of vecuronium were administered sequentially until blockade exceeded 90%. The degree of blockade and onset time after the initial 10 microg/kg of vecuronium were assessed, and doses required to exceed 90% blockade were recorded. In addition, effective doses of 50% and 95% for vecuronium were calculated from a single data point. Both types of MG patients showed increased sensitivity to vecuronium compared with non-MG patients. IMPLICATIONS: Hypothesizing that seronegative patients are as sensitive to vecuronium as seropositive patients, we assessed sensitivity in seropositive and seronegative myasthenia gravis (MG) patients and in non-MG patients. They were, indeed, both equally sensitive to vecuronium. PMID- 12088953 TI - Small-dose ketamine improves the postoperative state of depressed patients. AB - We investigated whether ketamine is suitable for depressed patients who had undergone orthopedic surgery. We studied 70 patients with major depression and 25 patients as the control (Group C). The depressed patients were divided randomly into two groups; patients in Group A (n = 35) were induced with propofol, fentanyl, and ketamine and patients in Group B (n = 35) were induced with propofol and fentanyl, and all patients were maintained with 1.5%-2.0% isoflurane plus nitrous oxide. The mean Hamilton Depression Rating (HDR) score was 12.7 +/- 5.4 for Group A and 12.3 +/- 6.0 for Group B 2 days before surgery and 9.9 +/- 4.1 for Group A and 14.4 +/- 3.8 for Group B 1 day after surgery. The HDR score in Group A 1 day after surgery was significantly (P < 0.05) lower than that in Group B. The HDR score in Group C was 4.2 +/- 1.7 2 days before surgery and 4.8 +/- 1.6 1 day after surgery. Depressed mood, suicidal tendencies, somatic anxiety, and hypochondriasis significantly decreased in Group A as compared with Group B. Postoperative pain scores in Group A at 8 and 16 h after the end of anesthesia were 26.6 +/- 8.7 and 24.9 +/- 8.2, respectively, which were significantly (P < 0.05) lower than 34.3 +/- 12.0 and 31.1 +/- 8.8 in Group B. In conclusion, small-dose ketamine improved the postoperative depressive state and relieved postoperative pain in depressed patients. IMPLICATIONS: NMDA receptor antagonists are reported to be effective for improving depression. It remains unclear whether ketamine, which is an NMDA receptor antagonist, postoperatively affects the psychological state in depressed patients. We investigated the effect of 1.0 mg/kg of ketamine on postoperative outcomes in depressed patients. PMID- 12088954 TI - Can succinylcholine be used safely in hyperkalemic patients? AB - The use of succinylcholine in hyperkalemic patients (serum potassium >5.5 mEq/L) is often viewed as relatively contraindicated, although there are no systematic data to define what preoperative potassium level is safe. We retrospectively reviewed more than 40,000 general anesthetics administered over 70 mo in which succinylcholine was given at the induction. This search yielded 38 patients with a preoperative potassium of 5.6 mEq/L or greater. Survival of the anesthetic was 100%, and no dysrhythmias or other major morbidity were documented upon manual review of the intraoperative automated record keeper charts or the patient medical records. These data allow a 95% confidence interval assessment of maximal risk for an event of 7.9%, which is not negligible, but which almost certainly grossly overestimates the risk for patients with moderately increased potassium levels. A prospective trial to definitively assess the safety margin for succinylcholine use in hyperkalemic patients would be difficult. Therefore, these data, taken in the context of a compelling case for rapid intubating conditions without long-term paralysis, suggest safety in succinylcholine use in patients with modest hyperkalemia. IMPLICATIONS: In a review of more than 40,000 general anesthetics in which succinylcholine was given at induction, 38 patients had a preoperative potassium of 5.6 mEq/L or greater. All patients survived the anesthetic with no dysrhythmias or other major morbidity documented. Succinylcholine may be appropriate and safe for use in certain patients with moderate hyperkalemia. PMID- 12088956 TI - The dose-related efficacy of diltiazem for enhancing diaphragmatic fatigability in dogs. AB - Nicardipine, a calcium channel blockade, enhances the production of diaphragmatic fatigue. We studied the dose-related effects of diltiazem, another calcium channel blockade, on diaphragmatic fatigability in dogs. Animals were divided into three groups of eight each. In each group, diaphragmatic fatigue was induced by intermittent supramaximal bilateral electrophrenic stimulation at a frequency of 20 Hz applied for 30 min. During this fatigue-producing period, Group I received no study drug, Group II was infused with diltiazem 0.1 mg. kg(-1). h( 1), and Group III was infused with diltiazem 0.5 mg. kg(-1). h(-1). We assessed diaphragmatic contractility by transdiaphragmatic pressure (Pdi). After the fatigue-producing period, in Group I, Pdi at low-frequency (20-Hz) stimulation decreased from baseline values (P < 0.05), whereas there was no change in Pdi at high-frequency (100-Hz) stimulation. In Groups II and III, with an infusion of diltiazem, Pdi at both stimuli decreased from baseline values (P < 0.05). The decrease in Pdi to each stimulus was more in Group III than in Group II (P < 0.05). We conclude that diltiazem causes a dose-related augmentation of fatigability in the diaphragm of dogs. IMPLICATIONS: Diaphragmatic muscle fatigue is implicated as a cause of respiratory failure. Diltiazem, a calcium channel blockade, enhances diaphragmatic fatigability in dogs in a dose-related manner. PMID- 12088955 TI - Sevoflurane increases glucose transport in skeletal muscle cells. AB - Sevoflurane activates phospholipase C and protein kinase C, leading to an increase in intracellular Ca(2+) concentration, which modulates glucose transport. We studied in vitro the effect of sevoflurane on the uptake of 2 deoxyglucose in rat skeletal muscle cells and the mechanism that modulates the glucose transport. Sevoflurane 0.8, 1.2, and 2.0 mM significantly increased glucose uptake from 13.1 +/- 1.2 pmol. h(-1). mg protein(-1) to 22.6 +/- 1.4, 32.1 +/- 1.8, and 37.4 +/-2.7 pmol. h(-1). mg protein(-1), respectively. Tyrphostin A-23 (a highly selective tyrosine kinase inhibitor) 1 and 10 nM significantly decreased the sevoflurane-stimulated glucose uptake from 32.1 +/- 1.8 to 25.8 +/- 1.1 and 15.2 +/- 1.7 pmol. h(-1). mg protein(-1), respectively. Genistein (a selective tyrosine kinase inhibitor) 1 and 10 nM also significantly decreased the sevoflurane- stimulated glucose uptake from 32.1 +/- 1.8 to 25.7 +/ 1.5 and 15.2 +/- 1.4 pmol. h(-1). mg protein(-1), respectively. The sevoflurane stimulated glucose uptake was decreased by 100 nM and 1 microM TMB-8 (an intracellular Ca(2+) antagonist), from 32.1 +/- 1.8 pmol. h(-1). mg protein(-1) to 25.6 +/- 3.3 and 20.3 +/- 1.6 pmol. h(-1). mg protein(-1), respectively. Staurosporine (a protein kinase C antagonist) 100 nM significantly decreased sevoflurane-stimulated glucose uptake to 26.1 +/- 1.5 pmol. h(-1). mg protein( 1). We conclude that sevoflurane increases glucose uptake in skeletal muscle cells and that the sevoflurane-stimulated glucose uptake was associated with tyrosine kinase, protein kinase C, and intracellular Ca(2+). IMPLICATIONS: Sevoflurane anesthesia has an inhibitory effect on insulin secretion. Glucose concentrations in plasma do not significantly change during sevoflurane anesthesia. Plasma glucose concentrations are affected by intracellular glucose metabolism. However, glucose transport into cells during sevoflurane anesthesia remains unclear. PMID- 12088957 TI - The efficacy and safety of transdermal scopolamine for the prevention of postoperative nausea and vomiting: a quantitative systematic review. AB - The role of scopolamine administered via transdermal therapeutic systems in the prevention of postoperative vomiting, nausea, and nausea and vomiting is unclear. We performed a systematic search for full reports of randomized comparisons of transdermal scopolamine with inactive control. Dichotomous data were extracted. In the meta-analysis, relative risks and numbers-needed-to-treat/harm were calculated with 95% confidence intervals (CI). In 23 trials, 979 patients received transdermal scopolamine, and 984 patients received placebo. Sensitivity analyses were performed using restricted data for truncated control event rates (40%-80%) and for large trials. With these data, the relative risks for postoperative vomiting (five reports), nausea (five reports), nausea and vomiting (eight reports), and rescue treatment (three reports) were 0.69 (95% CI, 0.58 0.82), 0.69 (95% CI, 0.54-0.87), 0.76 (95% CI, 0.66-0.88), and 0.68 (95% CI, 0.54 0.85), respectively. This means that of 100 patients who receive transdermal scopolamine, approximately 17 will not experience postoperative vomiting who would have done so had they all received a placebo. However, 18 of 100 patients will have visual disturbances, eight will report dry mouth, two will report dizziness, one will be classified as being agitated, and 1-13 patients who are prescribed transdermal scopolamine will not use it correctly. The timing of application does not alter efficacy. IMPLICATIONS: Of 100 patients who receive transdermal scopolamine, approximately 17 will not vomit in the postoperative period who would have done so had they all received a placebo. However, 18 of 100 patients will have visual disturbances, and eight will report dry mouth. Incorrect use further limits its efficacy. PMID- 12088958 TI - The efficacy of lafutidine in improving preoperative gastric fluid property: a comparison with ranitidine and rabeprazole. AB - IMPLICATIONS: Acid aspiration syndrome remains a potentially critical perioperative complication. We compared lafutidine, ranitidine, and rabeprazole for reduction of preoperative gastric fluid acidity and volume in elective surgery and found that these variables were minimized with a single morning dose of lafutidine 20 mg compared with ranitidine or rabeprazole. Preoperative oral lafutidine may be an alternative to ranitidine as a prophylaxis against aspiration pneumonia. PMID- 12088959 TI - E-cylinder-powered mechanical ventilation may adversely impact anesthetic management and efficiency. AB - Anesthesiologists often administer care outside the operating room. These locations may depend on gas cylinders for their oxygen source more than the operating suites supplied by dedicated central pipelines. Using full E-cylinders, we determined the oxygen consumption of two often used, pneumatically driven anesthesia ventilators to answer three questions: How much time is available when mechanically ventilating patients in the setting of absent or malfunctioning central oxygen pipeline? How much oxygen is used by the ventilator to drive the bellows? How does changing the inspiratory to expiratory ratio and the inspiratory flow (Narkomed ventilator only) influence oxygen use? At a ventilation of 5 L/min, we found that mechanical ventilation consumes between 59% and 85% of the available oxygen in an E-cylinder to drive the ventilator at fresh gas flows ranging from 1 to 10 L/min. The time span until the low oxygen alarm sounded ranged from 38 to 99 min. Alteration of the inspiratory flow but not the inspiratory to expiratory ratio had a significant impact. Clinicians must recognize that mechanical ventilation using E-cylinders rapidly depletes this sole oxygen source and could jeopardize patient safety. Conversely, manual or spontaneous ventilation with low fresh gas flows minimizes oxygen depletion. IMPLICATIONS: The time available to ventilate patients with an E-cylinder tank as the sole oxygen source was found to be as short as 38 min. Clinicians must recognize that mechanical ventilation using oxygen cylinders rapidly depletes oxygen and could jeopardize patient safety. PMID- 12088960 TI - A new highly reliable instrument for the assessment of pre- and postoperative gynecological pain. AB - In this study, we evaluated the reliability of a newly developed method for pain assessment, which is based on perceptual matching by Pain Matcher, Cefar Medical AB, Lund, Sweden, during minor gynecological surgery. In addition, the responsiveness to two different anesthetic methods-electro-acupuncture or a fast acting opiate, alfentanil, both in combination with a paracervical block-was estimated by using Pain Matcher and visual analog scale (VAS) assessments before and 2 h after surgery. Two hundred-twenty-three women (aged 22-38 yr) participated. The results show that Pain Matcher is a reliable method for pain assessments, with lack of random individual disagreement and with no statistical evidence of systematic disagreement in position or in concentration. The augmented rank-order coefficient (r(a)) values were excellent (0.95-1.00). When scales were used to detect true changes over time, there was no clear indication of responsiveness, mostly because of statistically significant random individual changes. However, the individual changes were much smaller for magnitude matching than for VAS. In conclusion, we would recommend the use of perceptual matching by Pain Matcher for pain assessment, because in this study it was a reliable and powerful in test-retest situations and had smaller individual changes than VAS after intervention. The Pain Matcher procedure was well accepted by the patients, and the results suggest that it may be useful when evaluating acute pre- and postoperative pain. IMPLICATIONS: We evaluated a new instrument for pain assessment. Our results show that this method is highly reliable, is well tolerated by the patients, is reported to be easy to use, and may be useful when evaluating acute pre- and postoperative pain. PMID- 12088961 TI - The analgesic effects of intraperitoneal and incisional bupivacaine with epinephrine after total abdominal hysterectomy. AB - The objective of our study was to see if incisional and intraperitoneal bupivacaine with epinephrine produces analgesia after total abdominal hysterectomy. Forty-six ASA physical status I and II patients received a standardized anesthetic, patient-controlled analgesia (PCA) morphine, and rectal paracetamol 1 g every 6 h. Patients were randomized to receive 50 mL of bupivacaine 0.25% with epinephrine 5 microg/mL or 50 mL of normal saline. Thirty milliliters and 20 mL of treatment solution were administered into the peritoneum and incision, respectively, before wound closure. Seventeen and 16 patients in the Placebo and Bupivacaine groups, respectively, completed the study. The reasons for withdrawal were PCA malfunction, PCA discontinued too early, nausea, chest infection, intraabdominal drain insertion, and protocol violation. There were no significant differences between the Bupivacaine and Placebo groups in age, height, weight, or duration of surgery. Pain on movement was significantly more intense in the Placebo group than in the Bupivacaine group on awakening. Morphine consumption (interquartile range) over 24 h was 62 mg (53-85 mg) in the Placebo group compared with 44 mg (33-56 mg) in the Bupivacaine group (P < 0.01). This significant difference was attributable to the larger morphine consumption in the Placebo group in the first 4 postoperative h. We conclude that a combination of intraperitoneal and incisional bupivacaine with epinephrine provides significant morphine-sparing analgesia for 4 h after total abdominal hysterectomy. IMPLICATIONS: A combination of intraperitoneal and incisional bupivacaine with epinephrine may be recommended because it provides significant morphine-sparing analgesia for 4 h after total abdominal hysterectomy. PMID- 12088962 TI - An investigation of monoamine receptors involved in antinociceptive effects of antidepressants. AB - We attempted to determine which monoamine receptor subtypes are predominantly involved in antidepressant-induced antinociception. Antinociceptive effects were evaluated by using formalin tests with rats. Antidepressants acting as potent inhibitors of norepinephrine reuptake (nisoxetine, nortriptyline, and maprotiline) or inhibiting reuptake of both norepinephrine and serotonin (5-HT) (imipramine and milnacipran) induced dose-dependent antinociception. Simultaneous intraperitoneal administration of antidepressants and either prazosin (alpha(1) antagonist) or ketanserin (5-HT(2) antagonist) significantly antagonized antinociceptive effects. Fluvoxamine (selective serotonin reuptake inhibitor) induced antinociception less potently than other antidepressants and was significantly antagonized by ketanserin, but not prazosin. Ondansetron (5-HT(3) antagonist) significantly antagonized antinociception by 10 mg/kg of imipramine. In contrast, SDZ-205,557 (5-HT(4) antagonist) markedly enhanced antinociception by small-dose (2.5 mg/kg) imipramine. Imipramine-induced antinociception was significantly antagonized by intracerebroventricular administration of prazosin or ketanserin, but not by yohimbine (alpha(2) antagonist) or ondansetron, and was significantly enhanced by intracerebroventricularly administered SDZ-205,557. These findings suggest that alpha(1) adrenoceptors and 5-HT(2) receptors in the brain are involved in antidepressant-induced antinociception. In addition, the results suggested functional interactions between noradrenergic and serotonergic neurons as mechanisms for antidepressant-induced antinociception. IMPLICATIONS: Formalin tests of rats treated with antidepressants and antagonists of monoamine receptors indicate that alpha(1) adrenoceptors, serotonin (5-HT)(2) receptors, and 5-HT(3) receptors are involved in antidepressant-induced antinociception, suggesting functional interactions between noradrenergic and serotonergic neurons as mechanisms of antidepressant-induced antinociception. PMID- 12088963 TI - Cyclooxygenase inhibition for postoperative analgesia. PMID- 12088964 TI - Anesthesiologists and acute perioperative stress: a cohort study. AB - Previous studies have indicated that many anesthesiologists exhibit symptoms of chronic stress. There is a paucity of data, however, regarding the existence of acute stress signs among anesthesiologists. Anesthesiologists from three practice settings (n = 38) were studied while they were anesthetizing 203 patients. Heart rate (HR) was recorded continuously and arterial blood pressure (BP) was measured hourly and immediately after each induction. Anxiety levels and salivary cortisol levels were also assessed after each induction. Comparison BP and HR data were obtained from the anesthesiologists during a nonclinical day. We found that anesthesiologists' HR increased during the anesthetic process compared with morning baseline HR (P = 0.008). This HR increase, however, was not clinically significant; the average HR during the anesthetic pro- cess ranged from 80 +/- 12 to 84 +/- 11 bpm. Similarly, although both systolic and diastolic BP after inductions were increased compared with baseline BP (P = 0.001), this increase was not clinically significant. In 9% of the inductions, however, systolic BP exceeded 140 mm Hg, and in 17% of all inductions, diastolic BP exceeded 90 mm Hg. Finally, the average BP of anesthesiologists during a clinical day was not different from the average BP during a nonclinical day (P = 0.9). Self-reported anxiety did not increase significantly after inductions (P = 0.15). An analysis of Holter tapes revealed no rhythm abnormalities and no signs of myocardial ischemia. We conclude that the practice of anesthesiology is associated with minor manifestations of acute physiologic stress during the perioperative process. IMPLICATIONS: Anesthesiologists experience minor psychologic stress while involved in the anesthetic process. PMID- 12088965 TI - Sampling error can significantly affect measured hospital financial performance of surgeons and resulting operating room time allocations. AB - Hospitals with limited operating room (OR) hours, those with intensive care unit or ward beds that are always full, or those that have no incremental revenue for many patients need to choose which surgeons get the resources. Although such decisions are based on internal financial reports, whether the reports are statistically valid is not known. Random error may affect surgeons' measured financial performance and, thus, what cases the anesthesiologists get to do and which patients get to receive care. We tested whether one fiscal year of surgeon specific financial data is sufficient for accurate financial accounting. We obtained accounting data for all outpatient or same-day-admit surgery cases during one fiscal year at an academic medical center. Linear programming was used to find the mix of surgeons' OR time allocations that would maximize the contribution margin or minimize variable costs. Confidence intervals were calculated on these end points by using Fieller's theorem and Monte-Carlo simulation. The 95% confidence intervals for increases in contribution margins or reductions in variable costs were 4.3% to 10.8% and 6.0% to 8.9%, respectively. As many as 22% of surgeons would have had OR time reduced because of sampling error. We recommend that physicians ask for and OR managers get confidence intervals of end points of financial analyses when making decisions based on them. IMPLICATIONS: The common approach of using one fiscal year of perioperative accounting data can be insufficient to prevent random error from influencing important management decisions. When accounting data are used for hospital and operating room management decision making, confidence intervals should be calculated for the key financial variables (e.g., variable cost per hour of operating room time). PMID- 12088966 TI - Electroventilation with monopolar and bipolar intratracheal electrodes. AB - Electroventilation is the technique of producing inspiration with stimuli applied to inspiratory muscles by using strategically placed electrodes on or within the body. We conducted studies using the dog to determine the efficacy of monopolar and bipolar intratracheal electrodes for electroventilation. Bipolar electrodes were able to produce an inspired volume greater than a spontaneous tidal volume in all seven dogs, but monopolar electrodes were able to in only three of seven dogs. The intratracheal electrode has the advantage over body-surface electrodes that less current is required to produce a single tidal volume, and it has the advantage over intraesophageal electrodes that anatomical placement of the electrode is simplified. These experiments demonstrated that a bipolar electrode can be used for electroventilation. IMPLICATIONS: This study demonstrates that electroventilation is possible with intratracheal electrodes, decreasing the current needed to electroventilate by using body-surface electrodes. Two different methods of intratracheal electroventilation can be used: monopolar, in which the return electrode is on the body surface; and bipolar, in which the return electrode is in the trachea. PMID- 12088967 TI - Modeling the effect of progressive endotracheal tube occlusion on tidal volume in pressure-control mode. AB - A recognized hazard of prolonged endotracheal intubation is progressive airway occlusion resulting from deposition of secretions on the inner surface of the endotracheal tube (ETT). When volume-controlled ventilation is used, progressive ETT occlusion may be detected by monitoring the difference between peak and plateau airway pressures. In pressure-controlled modes, however, inspiratory airway pressures are preset and thus cannot act as a warning indicator. Instead, changes in delivered tidal volumes may aid the diagnosis of ETT occlusion. To determine whether tidal volume monitoring effectively detects progressive ETT occlusion, we mathematically modeled the response of a ventilator operating in pressure-controlled mode to increasing airway resistance. To corroborate our model, we then bench-tested the Siemens 300 and Puritan-Bennett 7200 ventilators by using a test lung and a series of ETTs ranging in size from 9.0 to 3.5 mm inner diameter to simulate progressive occlusion. We found that when pressure controlled mode was used, progressive ETT occlusion did not reduce delivered tidal volumes until occlusion was nearly complete. We conclude that prolonged use of pressure-controlled mode may allow significant ETT obstruction to build up undetected, risking complete ETT occlusion and complicating the perioperative care of patients ventilated with this mode. IMPLICATIONS: Although increasing airway pressures during volume-controlled ventilation allow early recognition of endotracheal tube (ETT) obstruction, airway pressures with pressure-controlled ventilation are fixed. We found during tests of two intensive care unit ventilators that although ETT obstruction reduces delivered tidal volumes during pressure-controlled ventilation, reductions do not occur until occlusion is advanced. PMID- 12088968 TI - Does spinal anesthesia cause hearing loss in the obstetric population? AB - Lumbar puncture is believed to cause hypoacousis by causing cerebrospinal fluid leakage in older individuals. We hypothesized that parturients undergoing subarachnoid block (SAB) may experience hearing loss. We evaluated the effects of SAB on hearing in parturients undergoing elective cesarean delivery. We also compared two types of spinal needles: a pencil-point needle (24-gauge Sprotte needle) and a cutting needle (25-gauge Quincke needle). Sixty patients were studied: 20 received lumbar epidural block for labor analgesia (controls), 20 received a SAB with a Sprotte needle, and 20 others received a SAB with a Quincke needle for cesarean delivery. A tone audiometer was used to test for that decibel level at which the patient heard 125-, 250-, 500-, 1000-, 2000-, 4000-, and 8000 Hz frequencies. The hearing test was performed before anesthesia, after delivery, and on the first and second postoperative days. The results were analyzed by using repeated-measures analysis of variance at P < 0.05. No patient from any of the three groups developed a hearing loss either at low or high frequencies. Spinal anesthesia does not lead to significant hearing loss when a pencil- or a cutting-point needle is used in the obstetric population. IMPLICATIONS: Sixty obstetric patients were enrolled in the study to examine the possible effects of spinal anesthesia on their hearing. By using an audiometer, the patient's hearing was evaluated before delivery, after delivery, and for the following 2 days. There was no significant change of hearing in any of the patients. PMID- 12088969 TI - The relative motor blocking potencies of epidural bupivacaine and ropivacaine in labor. AB - Minimal local analgesic concentrations (MLAC) have been used to determine the epidural analgesic potencies of bupivacaine and ropivacaine. There are no reports of the motor blocking potencies of these drugs. We sought to determine the motor block MLAC of both drugs and their relative potency ratio. Sixty ASA physical status I and II parturients were randomized to one of two groups, during the first stage of labor. Each received a 20-mL bolus of epidural bupivacaine or ropivacaine. The first woman in each group received 0.35%. Up-down sequential allocation was used to determine subsequent concentrations at a testing interval of 0.025%. Effective motor block was defined as a Bromage score <4 within 30 min. The up-down sequences were analyzed by using the Dixon and Massey method and probit regression to quantify the motor block minimal local analgesic concentration. Two-sided P < 0.05 defined significance. The motor block minimal local analgesic concentration for bupivacaine was 0.326% (95% confidence interval [CI], 0.285-0.367) and for ropivacaine was 0.497% (95% CI, 0.431-0.563) (P = 0.0008). The ropivacaine/bupivacaine potency ratio was 0.66 (95% CI, 0.52-0.82). This is the first MLAC study to estimate the motor blocking potencies of bupivacaine and ropivacaine. Ropivacaine was significantly less potent for motor block, at 66% that of bupivacaine. IMPLICATIONS: The results of this study demonstrate that epidural ropivacaine is less potent than epidural bupivacaine in producing motor blockade during labor. The motor block potency relation is similar to the sensory potency ratio for these two drugs. PMID- 12088971 TI - New landmarks for the anterior approach to the sciatic nerve block: imaging and clinical study. AB - In this study, we assessed the reliability of the inguinal crease and femoral artery as anatomic landmarks for the anterior approach to the sciatic nerve and determined the optimal position of the leg during this approach. An imaging study was conducted before the clinical study. The sciatic nerve was located twice in 20 patients undergoing ankle or foot surgery, once with the leg in the neutral position and once with the leg in the externally rotated position. The patient was lying supine. A 22-gauge, 150-mm insulated b-beveled needle connected to a nerve stimulator was inserted 2.5 cm distal to the inguinal crease and 2.5 cm medial to the femoral artery and was directed posteriorly and laterally with a 10 degrees -15 degrees angle relative to the vertical plane. The sciatic nerve was located in all patients at a depth of 10.6 +/- 1.8 cm when the leg was in the neutral position and 10.4 +/- 1.5 cm when the leg was in the externally rotated position (not significant). In the neutral position and in the externally rotated position, the time needed to identify anatomic landmarks was 28 +/- 15 s and 26 +/- 14 s, respectively (not significant), and the time needed to locate the sciatic nerve was 79 +/- 53 s and 46 +/- 25 s (P < 0.006), respectively. We conclude that the inguinal crease and femoral artery are reliable and effective anatomic landmarks for the anterior approach to the sciatic nerve and that the optimal position of the leg is the externally rotated position. IMPLICATIONS: This new anterior approach to the sciatic nerve using the inguinal crease and femoral artery as landmarks is an easy and reliable technique. PMID- 12088970 TI - Intrathecal versus intravenous fentanyl for supplementation of subarachnoid block during cesarean delivery. AB - Forty-eight healthy parturients scheduled for elective cesarean delivery were randomly allocated to receive intrathecally either 12 mg of hyperbaric bupivacaine plus 12.5 microg of fentanyl (n = 23) or bupivacaine alone (n = 25). In the latter group, IV 12.5 microg of fentanyl was administered immediately after spinal anesthesia. We compared the amount of IV fentanyl required for supplementation of the spinal anesthesia during surgery, the intraoperative visual analog scale, the time to the first request for postoperative analgesia, and the incidence of adverse effects. Additional IV fentanyl supplementation amounting to a mean of 32 +/- 35 microg was required in the IV Fentanyl group, whereas no supple- mentation was required in the Intrathecal Fentanyl group (P = 0.009). The time to the first request for postoperative analgesia was significantly longer in the Intrathecal Fentanyl group than in the IV Fentanyl group (159 +/- 39 min versus 119 +/- 44 min; P = 0.003). The incidence of systolic blood pressure <90 mm Hg and the ephedrine requirements were significantly higher in the IV Fentanyl group as compared with the Intrathecal Fentanyl group (P = 0.01). Also, intraoperative nausea and vomiting occurred less frequently in the Intrathecal Fentanyl group compared with the IV Fentanyl group (8 of 23 vs 17 of 25; P = 0.02). IMPLICATIONS: Supplementation of spinal bupivacaine anesthesia for cesarean delivery with intrathecal fentanyl provides a better quality of anesthesia and is associated with a decreased incidence of side effects as compared with supplementation with the same dose of IV fentanyl. PMID- 12088972 TI - Levobupivacaine for axillary brachial plexus block: a pharmacokinetic and clinical comparison in patients with normal renal function or renal disease. AB - We compared the pharmacokinetics and clinical characteristics of 0.5% levobupivacaine for axillary block in patients with normal renal function versus patients with end-stage renal disease (ESRD). Twenty patients with normal renal function and eight patients with ESRD received an axillary block with 50-60 mL of 0.5% levobupivacaine. Patients were evaluated for onset and duration of sensory/motor block. Eleven patients with normal renal function and eight patients with ESRD underwent pharmacokinetic analysis. No differences between groups were found in the onset, duration, or quality of block. The median time to sensory block was 12.5 min and 12.9 min, and mean duration of the block was 19 h and 22 h in normal versus ESRD patients, respectively. No significant differences in noncompartmental pharmacokinetic variables (median) were found between normal and ESRD patients with an AUC(0-t) (microg. h(-1). mL(-1)) of 11 and 13, peak concentration (C(max)) (microg/mL) of 1.2 and 1.6, and a time to peak concentration (T(max)) (min) of 55 and 48, respectively. This study demonstrates the clinical efficacy and equivalence of the pharmacokinetic characteristics of 0.5% levobupivacaine for axillary brachial plexus block in patients with ESRD and normal renal function. IMPLICATIONS: This study demonstrates the clinical efficacy and equivalence of the pharmacokinetic characteristics of 0.5% levobupivacaine for axillary brachial plexus block in patients with renal disease and normal renal function. PMID- 12088973 TI - Temporary bilateral blindness after acute lidocaine toxicity. AB - IMPLICATIONS: This case report describes an uncommon complication (blindness) occurring after an inadvertent overdosage of a frequently used local anesthetic (lidocaine) during a regional anesthetic procedure. The discussion focuses on the suspected pathophysiology of the blindness. PMID- 12088974 TI - The resolution of ST segment depressions after high right thoracic paravertebral block during general anesthesia. AB - IMPLICATIONS: Thoracic epidural, stellate ganglion, and thoracic paravertebral blocks all relieve angina. We report a case of intraoperative resolution of ST segment depression after a right thoracic paravertebral block. PMID- 12088975 TI - Small-dose dexamethasone reduces nausea and vomiting after laparoscopic cholecystectomy: a comparison of tropisetron with saline. AB - Dexamethasone is an effective antiemetic drug, but the efficacy of small-dose dexamethasone 5 mg on the prophylaxis of postoperative nausea and vomiting (PONV) in patients undergoing laparoscopic cholecystectomy has not been evaluated. We, therefore, evaluated the prophylactic effect of small-dose dexamethasone (5 mg) on PONV in patients undergoing laparoscopic cholecystectomy. Tropisetron and saline served as controls. One-hundred-twenty patients scheduled for laparoscopic cholecystectomy were enrolled in a randomized, double-blinded, placebo-controlled study. At the induction of anesthesia, the Dexamethasone group received IV dexamethasone 5 mg, the Tropisetron group received IV tropisetron 2 mg, and the Placebo group received IV saline. We found that both dexamethasone and tropisetron significantly decreased the following variables: the total incidence of PONV (P < 0.01), more than four vomiting episodes (P < 0.05), and the proportions of patients requiring rescue antiemetics (P < 0.05). The differences between the Dexamethasone and Tropisetron groups were not significant. We conclude that prophylactic IV dexamethasone 5 mg significantly reduces the incidence of PONV in patients undergoing laparoscopic cholecystectomy. At this dose, dexamethasone is as effective as tropisetron 2 mg and is more effective than placebo. IMPLICATIONS: We evaluated the prophylactic effect of small-dose dexamethasone (5 mg) on postoperative nausea and vomiting (PONV) in patients undergoing laparoscopic cholecystectomy. Tropisetron (2 mg) and saline served as controls. We found that dexamethasone 5 mg (IV) significantly reduced the incidence of PONV in these patients, and, at this dose, dexamethasone was as effective as tropisetron and was more effective than placebo. PMID- 12088976 TI - Fentanyl attenuates the hemodynamic response to endotracheal intubation more than the response to laryngoscopy. AB - We examined the effectiveness of avoiding laryngoscopy in reducing the hemodynamic responses to orotracheal intubation during the induction of anesthesia. One hundred surgical patients who required orotracheal intubation were randomly allocated into four groups. The first and third groups underwent fiberoptic intubation, in which an anesthesiologist inserted the endotracheal tube into the trachea under TV monitoring through a bronchoscope, and the second and fourth groups underwent conventional orotracheal intubation using a rigid laryngoscope. The third and fourth groups were pretreated with 2 microg/kg fentanyl IV immediately before the induction of anesthesia. Blood pressure and heart rate were measured noninvasively. A significant reduction in hemodynamic response was seen in only the group treated with fentanyl and intubated using the fiberoptic technique. Without fentanyl, there was no significant difference in hemodynamic changes between the groups. We conclude that the administration of fentanyl suppresses the hemodynamic responses to endotracheal intubation more than it does to laryngoscopy. There was no significant difference in the hemodynamic responses to orotracheal intubation by fiberscopy and laryngoscopy without fentanyl pretreatment, whereas 2 microg/kg fentanyl significantly reduced the hemodynamic responses in the group intubated by fiberscopy. IMPLICATIONS: We assessed the effectiveness of avoiding laryngoscopy for orotracheal intubation. There was no significant difference in the hemodynamic responses to orotracheal intubation by fiberscopy and laryngoscopy without fentanyl pretreatment, whereas 2 microg/kg fentanyl significantly reduced the hemodynamic responses in the group intubated by fiberscopy. Pretreatment of fentanyl and fiberoptic intubation might be recommended for avoiding hyperdynamic responses. PMID- 12088977 TI - Rapid deflation of the bronchial cuff of the double-lumen tube after decreasing the concentration of inspired nitrous oxide. AB - Deflationary phenomena of the endotracheal tube cuff may occur after inspired nitrous oxide (N(2)O) concentrations are reduced, but deflationary phenomena of the double-lumen tube (DLT) cuff have not been investigated. In this study, tracheal and bronchial cuffs of left-sided Mallinckrodt (Athlone, Ireland) DLTs were inflated with air, 40% N(2)O, or 67% N(2)O (Air, N40, or N67 groups, respectively) in 24 patients undergoing thoracic surgery; 40 min later, O(2) was substituted for N(2)O in some of the patients in the N40 group (N40-c group). Intracuff gas volumes, N(2)O concentrations, and cuff compliance were also measured. Both tracheal and bronchial cuff pressures significantly increased in the Air group but decreased in the N67 group. Neither pressure significantly changed in the N40 group, but both decreased in the N40-c group after terminating N(2)O anesthesia; the time required for bronchial cuff pressures to decrease by half (12.0 +/- 5.5 min) was less than that for tracheal cuff pressures (31.2 +/- 11.0 min, P < 0.01). The volume change in the N40-c group was not significantly different between the tracheal and bronchial cuffs, but tracheal cuff compliance was significantly higher than bronchial compliance. Therefore, filling DLT cuffs with 40% N(2)O stabilizes cuff pressure during anesthesia with 67% N(2)O, but bronchial cuffs deflate more quickly than tracheal cuffs after cessation of N(2)O administration through smaller compliance. IMPLICATIONS: We demonstrated that after cessation of nitrous oxide (N(2)O) administration, bronchial N(2)O-filled cuffs of the double-lumen tube deflate more rapidly than tracheal cuffs. To avoid insufficient separation of the lungs by the bronchial cuff, a frequent check of the cuff pressure is recommended after the inspired N(2)O concentration is decreased. PMID- 12088978 TI - Repeated deflation of a gas-barrier cuff to stabilize cuff pressure during nitrous oxide anesthesia. AB - Although a nitrous oxide (N(2)O) gas-barrier cuff effectively limits the increase of cuff pressure during N(2)O anesthesia, there are few data assessing whether an N(2)O gas-barrier cuff is more beneficial for stabilizing intracuff pressure than standard endotracheal tubes when cuffs are repeatedly deflated to stabilize pressure during N(2)O anesthesia. In the present study, the pressure of air filled standard-type cuffs (Trachelon; Terumo, Tokyo, Japan) and N(2)O gas barrier type endotracheal tube cuffs (Profile Soft-Seal Cuff [PSSC]; Sims Portex, Kent, UK) was measured during 67% N(2)O anesthesia (n = 8 in each), during which the cuffs were repeatedly deflated every 30 min (Trachelon) or 60 min (PSSC) for the first 3 or 4 h. After aspirating the cuffs for 3 h, the cuff pressure exceeded 22 mm Hg in more than half of the patients in both groups. However, aspiration of the cuffs for 4 h decreased the maximal cuff pressure between deflation intervals in both groups (P < 0.01 for each), and increased the intracuff N(2)O concentration (P < 0.0001 for each). After deflating the cuffs over 4 h, the cuff pressure in both groups never exceeded 22 mm Hg during the subsequent 3 h, and intracuff N(2)O concentrations did not significantly change. Therefore, deflation of cuffs for 4 h during N(2)O anesthesia sufficiently stabilized cuff pressure and equilibrated the intracuff N(2)O concentrations in both groups. The use of the PSSC endotracheal tube might be more practical because of the smaller number of cuff deflations required, but the PSSC does not reduce the duration of cuff deflations to stabilize the pressure. IMPLICATIONS: We demonstrated that the N(2)O concentration and pressure in the N(2)O-barrier Profile Soft-Seal Cuff stabilized when the cuff was aspirated once an hour for 4 h during N(2)O anesthesia. The Profile Soft-Seal Cuff might be easier to use in clinical practice than standard endotracheal tubes because of the smaller number of cuff deflations required. PMID- 12088979 TI - The use of the intubating laryngeal mask endotracheal tube with intubating devices. AB - IMPLICATIONS: Despite adequate visualization of the vocal cords using specialized airway devices, anatomical factors and the physical characteristics of the tube may cause difficulty when performing endotracheal intubation. The endotracheal tube designed for use with the intubating laryngeal mask airway may facilitate intubation in these circumstances. PMID- 12088980 TI - Physiological or functional fluid spaces. PMID- 12088981 TI - Effectiveness of blinding when comparing nitrous oxide with topical anesthetic cream. PMID- 12088982 TI - Popliteal block as an alternative to Labat's approach. PMID- 12088983 TI - Did sevoflurane contribute to hepatic failure in a child given large doses of acetaminophen? PMID- 12088984 TI - Hemostasis in patients of different ABO blood groups. PMID- 12088985 TI - What to do with clonidine with long-acting local anesthetic in brachial plexus block? PMID- 12088986 TI - Can BIS monitoring delay the development of clinical judgment? PMID- 12088987 TI - Myocardial ischemia during emergency anesthesia in a patient with systemic lupus erythematosus resulting from undiagnosed antiphospholipid syndrome. PMID- 12088988 TI - Acute oxygen desaturation and right heart dysfunction secondary to transesophageal echocardiography-induced malpositioning of the endotracheal tube. PMID- 12088989 TI - In response to "A multimodal postoperative rehabilitation program in critically ill diabetic patients". PMID- 12088990 TI - Bicarbonate administration is potentially harmful in patients with moderate acidosis. PMID- 12088992 TI - Knotting of an epidural catheter like a tie. PMID- 12088993 TI - Characterization of a Listeria monocytogenes Scott A isolate with high tolerance towards high hydrostatic pressure. AB - An isolate of L. monocytogenes Scott A that is tolerant to high hydrostatic pressure (HHP), named AK01, was isolated upon a single pressurization treatment of 400 MPa for 20 min and was further characterized. The survival of exponential- and stationary-phase cells of AK01 in ACES [N-(2-acetamido)-2-aminoethanesulfonic acid] buffer was at least 2 log units higher than that of the wild type over a broad range of pressures (150 to 500 MPa), while both strains showed higher HHP tolerance (piezotolerance) in the stationary than in the exponential phase of growth. In semiskim milk, exponential-phase cells of both strains showed lower reductions upon pressurization than in buffer, but again, AK01 was more piezotolerant than the wild type. The piezotolerance of AK01 was retained for at least 40 generations in rich medium, suggesting a stable phenotype. Interestingly, cells of AK01 lacked flagella, were elongated, and showed slightly lower maximum specific growth rates than the wild type at 8, 22, and 30 degrees C. Moreover, the piezotolerant strain AK01 showed increased resistance to heat, acid, and H(2)O(2) compared with the wild type. The difference in HHP tolerance between the piezotolerant strain and the wild-type strain could not be attributed to differences in membrane fluidity, since strain AK01 and the wild type had identical in situ lipid melting curves as determined by Fourier transform infrared spectroscopy. The demonstrated occurrence of a piezotolerant isolate of L. monocytogenes underscores the need to further investigate the mechanisms underlying HHP resistance of food-borne microorganisms, which in turn will contribute to the appropriate design of safe, accurate, and feasible HHP treatments. PMID- 12088994 TI - Secondary (gamma-Proteobacteria) endosymbionts infect the primary (beta Proteobacteria) endosymbionts of mealybugs multiple times and coevolve with their hosts. AB - Mealybugs (Hemiptera, Coccoidea, Pseudococcidae) are plant sap-sucking insects that have within their body cavities specialized cells containing prokaryotic primary endosymbionts (P-endosymbionts). The P-endosymbionts have the unusual property of containing within their cytoplasm prokaryotic secondary endosymbionts (S-endosymbionts) [C. D. von Dohlen, S. Kohler, S. T. Alsop, and W. R. McManus, Nature (London) 412:433-436, 2001]. Four-kilobase fragments containing 16S-23S ribosomal DNA (rDNA) were obtained from the P-endosymbionts of 22 mealybug species and the S-endosymbionts of 12 representative species. Phylogenetic analyses of the P-endosymbionts indicated that they have a monophyletic origin and are members of the beta-subdivision of the Proteobacteria; these organisms were subdivided into five different clusters. The S-endosymbionts were members of the gamma-subdivision of the Proteobacteria and were grouped into clusters similar to those observed with the P-endosymbionts. The S-endosymbiont clusters were distinct from each other and from other insect-associated bacteria. The similarity of the clusters formed by the P- and S-endosymbionts suggests that the P-endosymbionts of mealybugs were infected multiple times with different precursors of the S-endosymbionts and once the association was established, the P and S-endosymbionts were transmitted together. The lineage consisting of the P endosymbionts of mealybugs was given the designation "Candidatus Tremblaya" gen. nov., with a single species, "Candidatus Tremblaya princeps" sp. nov. The results of phylogenetic analyses of mitochondrial DNA fragments encoding cytochrome oxidase subunits I and II from four representative mealybug species were in agreement with the results of 16S-23S rDNA analyses, suggesting that relationships among strains of "Candidatus T. princeps" are useful in inferring the phylogeny of their mealybug hosts. PMID- 12088995 TI - The genetic properties of the primary endosymbionts of mealybugs differ from those of other endosymbionts of plant sap-sucking insects. AB - Mealybugs (Hemiptera, Coccoidea, Pseudococcidae), like aphids and psyllids, are plant sap-sucking insects that have an obligate association with prokaryotic endosymbionts that are acquired through vertical, maternal transmission. We sequenced two fragments of the genome of Tremblaya princeps, the endosymbiont of mealybugs, which is a member of the beta subdivision of the Proteobacteria. Each of the fragments (35 and 30 kb) contains a copy of 16S-23S-5S rRNA genes. A total of 37 open reading frames were detected, which corresponded to putative rRNA proteins, chaperones, and enzymes of branched-chain amino acid biosynthesis, DNA replication, protein translation, and RNA synthesis. The genome of T. princeps has a number of properties that distinguish it from the genomes of Buchnera aphidicola and Carsonella ruddii, the endosymbionts of aphids and psyllids, respectively. Among these properties are a high G+C content (57.1 mol%), the same G+C content in intergenic spaces and structural genes, and similar G+C contents of the genes encoding highly and poorly conserved proteins. The high G+C content has a substantial effect on protein composition; about one-third of the residues consist of four amino acids with high-G+C-content codons. Sequence analysis of DNA fragments containing the rRNA operon and adjacent regions from endosymbionts of several mealybug species suggested that there was a single duplication of the rRNA operon and the adjacent genes in an ancestor of the present T. princeps. Subsequently, in one mealybug lineage rpS15, one of the duplicated genes, was retained, while in another lineage it decayed. These results extend the diversity of the types of endosymbiotic associations found in plant sap-sucking insects. PMID- 12088996 TI - Members of the family Comamonadaceae as primary poly(3-hydroxybutyrate-co-3 hydroxyvalerate)-degrading denitrifiers in activated sludge as revealed by a polyphasic approach. AB - The distribution and phylogenetic affiliations of poly(3-hydroxybutyrate-co-3 hydroxyvalerate) (PHBV)-degrading denitrifying bacteria in activated sludge were studied by a polyphasic approach including culture-independent biomarker and molecular analyses as well as cultivation methods. A total of 23 strains of PHBV degrading denitrifiers were isolated from activated sludges from different sewage treatment plants. 16S ribosomal DNA (rDNA) sequence comparisons showed that 20 of the isolates were identified as members of the family Comamonadaceae, a major group of beta-Proteobacteria. When the sludges from different plants were acclimated with PHBV under denitrifying conditions in laboratory scale reactors, the nitrate removal rate increased linearly during the first 4 weeks and reached 20 mg NO(3)(-)-N h(-1) g of dry sludge(-1) at the steady state. The bacterial community change in the laboratory scale sludges during the acclimation was monitored by rRNA-targeted fluorescence in situ hybridization and quinone profiling. Both approaches showed that the population of beta-Proteobacteria in the laboratory sludges increased sharply during acclimation regardless of their origins. 16S rDNA clone libraries were constructed from two different acclimated sludges, and a total of 37 clones from the libraries were phylogenetically analyzed. Most of the 16S rDNA clones were grouped with members of the family Comamonadaceae. The results of our polyphasic approach indicate that beta Proteobacteria, especially members of the family Comamonadaceae, are primary PHBV degrading denitrifiers in activated sludge. Our data provide useful information for the development of a new nitrogen removal system with solid biopolymer as an electron donor. PMID- 12088997 TI - Parallel characterization of anaerobic toluene- and ethylbenzene-degrading microbial consortia by PCR-denaturing gradient gel electrophoresis, RNA-DNA membrane hybridization, and DNA microarray technology. AB - A mesophilic toluene-degrading consortium (TDC) and an ethylbenzene-degrading consortium (EDC) were established under sulfate-reducing conditions. These consortia were first characterized by denaturing gradient gel electrophoresis (DGGE) fingerprinting of PCR-amplified 16S rRNA gene fragments, followed by sequencing. The sequences of the major bands (T-1 and E-2) belonging to TDC and EDC, respectively, were affiliated with the family Desulfobacteriaceae. Another major band from EDC (E-1) was related to an uncultured non-sulfate-reducing soil bacterium. Oligonucleotide probes specific for the 16S rRNAs of target organisms corresponding to T-1, E-1, and E-2 were designed, and hybridization conditions were optimized for two analytical formats, membrane and DNA microarray hybridization. Both formats were used to characterize the TDC and EDC, and the results of both were consistent with DGGE analysis. In order to assess the utility of the microarray format for analysis of environmental samples, oil contaminated sediments from the coast of Kuwait were analyzed. The DNA microarray successfully detected bacterial nucleic acids from these samples, but probes targeting specific groups of sulfate-reducing bacteria did not give positive signals. The results of this study demonstrate the limitations and the potential utility of DNA microarrays for microbial community analysis. PMID- 12088998 TI - Differential ability of genotypes of 2,4-diacetylphloroglucinol-producing Pseudomonas fluorescens strains to colonize the roots of pea plants. AB - Indigenous populations of 2,4-diacetylphloroglucinol (2,4-DAPG)-producing fluorescent Pseudomonas spp. that occur naturally in suppressive soils are an enormous resource for improving biological control of plant diseases. Over 300 isolates of 2,4-DAPG-producing fluorescent Pseudomonas spp. were isolated from the rhizosphere of pea plants grown in soils that had undergone pea or wheat monoculture and were suppressive to Fusarium wilt or take-all, respectively. Representatives of seven genotypes, A, D, E, L, O, P, and Q, were isolated from both soils and identified by whole-cell repetitive sequence-based PCR (rep-PCR) with the BOXA1R primer, increasing by three (O, P, and Q) the number of genotypes identified previously among a worldwide collection of 2,4-DAPG producers. Fourteen isolates representing eight different genotypes were tested for their ability to colonize the rhizosphere of pea plants. Population densities of strains belonging to genotypes D and P were significantly greater than the densities of other genotypes and remained above log 6.0 CFU (g of root)(-1) over the entire 15-week experiment. Genetic profiles generated by rep-PCR or restriction fragment length polymorphism analysis of the 2,4-DAPG biosynthetic gene phlD were predictive of the rhizosphere competence of the introduced 2,4 DAPG-producing strains. PMID- 12088999 TI - Airborne microbial flora in a cattle feedlot. AB - A total of 1,408 cattle held in eight commercial feedlot pens were used to examine the quantity and diversity of microorganisms in cattle feedlot air. The effect of two feeding patterns on the generation of airborne dust and the total numbers of microorganisms was also examined (four feedlot pens/treatment). Microbial samples were collected, and dust particles that were 2.5 microm or less in diameter were measured with a Dustrak monitor during the evening dust peak for 4 days at sites both upwind and downwind of the feedlot pens. An Andersen biological cascade sampler was employed with different medium and incubation combinations for the capture and identification of bacteria and fungi. The results showed that when bacteria were considered, only nonpathogenic gram positive organisms were recovered. However, gram-negative bacteria may have been present in a viable but nonculturable state. Fungi were recovered in smaller numbers than bacteria, and none of the fungi were pathogenic. The Dustrak results showed that one feeding pattern resulted in cattle behavior that generated levels of downwind dust lower (P = 0.04) than the levels generated by the behavior resulting from the other feeding pattern. However, the Andersen sampler results showed that there were no differences between feeding patterns with regard to the total number or diversity of microorganisms. The disparity may have been due to the different operating principles of the two systems. The overall numbers of microorganisms recovered were lower than those reported in studies of intensively housed farm animals in which similar recovery techniques were used. PMID- 12089000 TI - Analysis of bacterial community composition by oligonucleotide fingerprinting of rRNA genes. AB - One of the first steps in characterizing an ecosystem is to describe the organisms inhabiting it. For microbial studies, experimental limitations have hindered the ability to depict diverse communities. Here we describe oligonucleotide fingerprinting of rRNA genes (OFRG), a method that permits identification of arrayed rRNA genes (rDNA) through a series of hybridization experiments using small DNA probes. To demonstrate this strategy, we examined the bacteria inhabiting two different soils. Analysis of 1,536 rDNA clones revealed 766 clusters grouped into five major taxa: Bacillus, Actinobacteria, Proteobacteria, and two undefined assemblages. Soil-specific taxa were identified and then independently confirmed through cluster-specific PCR of the original soil DNA. Near-species-level resolution was obtained by this analysis as clones with average sequence identities of 97% were grouped in the same cluster. A comparison of these OFRG results with the results obtained in a denaturing gradient gel electrophoresis analysis of the same two soils demonstrated the significance of this methodological advance. OFRG provides a cost-effective means to extensively analyze microbial communities and should have applications in medicine, biotechnology, and ecosystem studies. PMID- 12089002 TI - Functional production and characterization of a fibrin-specific single-chain antibody fragment from Bacillus subtilis: effects of molecular chaperones and a wall-bound protease on antibody fragment production. AB - To develop an ideal blood clot imaging and targeting agent, a single-chain antibody (SCA) fragment based on a fibrin-specific monoclonal antibody, MH-1, was constructed and produced via secretion from Bacillus subtilis. Through a systematic study involving a series of B. subtilis strains, insufficient intracellular and extracytoplasmic molecular chaperones and high sensitivity to wall-bound protease (WprA) were believed to be the major factors that lead to poor production of MH-1 SCA. Intracellular and extracytoplasmic molecular chaperones apparently act in a sequential manner. The combination of enhanced coproduction of both molecular chaperones and wprA inactivation leads to the development of an engineered B. subtilis strain, WB800HM[pEPP]. This strain allows secretory production of MH-1 SCA at a level of 10 to 15 mg/liter. In contrast, with WB700N (a seven-extracellular-protease-deficient strain) as the host, no MH-1 SCA could be detected in both secreted and cellular fractions. Secreted MH-1 SCA from WB800HM[pMH1, pEPP] could be affinity purified using a protein L matrix. It retains comparable affinity and specificity as the parental MH-1 monoclonal antibody. This expression system can potentially be applied to produce other single-chain antibody fragments, especially those with folding and protease sensitivity problems. PMID- 12089001 TI - Molecular characterization of the S-layer gene, sbpA, of Bacillus sphaericus CCM 2177 and production of a functional S-layer fusion protein with the ability to recrystallize in a defined orientation while presenting the fused allergen. AB - The nucleotide sequence encoding the crystalline bacterial cell surface (S-layer) protein SbpA of Bacillus sphaericus CCM 2177 was determined by a PCR-based technique using four overlapping fragments. The entire sbpA sequence indicated one open reading frame of 3,804 bp encoding a protein of 1,268 amino acids with a theoretical molecular mass of 132,062 Da and a calculated isoelectric point of 4.69. The N-terminal part of SbpA, which is involved in anchoring the S-layer subunits via a distinct type of secondary cell wall polymer to the rigid cell wall layer, comprises three S-layer-homologous motifs. For screening of amino acid positions located on the outer surface of the square S-layer lattice, the sequence encoding Strep-tag I, showing affinity to streptavidin, was linked to the 5' end of the sequence encoding the recombinant S-layer protein (rSbpA) or a C-terminally truncated form (rSbpA(31-1068)). The deletion of 200 C-terminal amino acids did not interfere with the self-assembly properties of the S-layer protein but significantly increased the accessibility of Strep-tag I. Thus, the sequence encoding the major birch pollen allergen (Bet v1) was fused via a short linker to the sequence encoding the C-terminally truncated form rSpbA(31-1068). Labeling of the square S-layer lattice formed by recrystallization of rSbpA(31 1068)/Bet v1 on peptidoglycan-containing sacculi with a Bet v1-specific monoclonal mouse antibody demonstrated the functionality of the fused protein sequence and its location on the outer surface of the S-layer lattice. The specific interactions between the N-terminal part of SbpA and the secondary cell wall polymer will be exploited for an oriented binding of the S-layer fusion protein on solid supports to generate regularly structured functional protein lattices. PMID- 12089003 TI - Monocyclic aromatic hydrocarbon degradation by Rhodococcus sp. strain DK17. AB - Rhodococcus sp. strain DK17 was isolated from soil and analyzed for the ability to grow on o-xylene as the sole carbon and energy source. Although DK17 cannot grow on m- and p-xylene, it is capable of growth on benzene, phenol, toluene, ethylbenzene, isopropylbenzene, and other alkylbenzene isomers. One UV-generated mutant strain, DK176, simultaneously lost the ability to grow on o-xylene, ethylbenzene, isopropylbenzene, toluene, and benzene, although it could still grow on phenol. The mutant strain was also unable to oxidize indole to indigo following growth in the presence of o-xylene. This observation suggests the loss of an oxygenase that is involved in the initial oxidation of the (alkyl)benzenes tested. Another mutant strain, DK180, isolated for the inability to grow on o xylene, retained the ability to grow on benzene but was unable to grow on alkylbenzenes due to loss of a meta-cleavage dioxygenase needed for metabolism of methyl-substituted catechols. Further experiments showed that DK180 as well as the wild-type strain DK17 have an ortho-cleavage pathway which is specifically induced by benzene but not by o-xylene. These results indicate that DK17 possesses two different ring-cleavage pathways for the degradation of aromatic compounds, although the initial oxidation reactions may be catalyzed by a common oxygenase. Gas chromatography-mass spectrometry and 300-MHz proton nuclear magnetic resonance spectrometry clearly show that DK180 accumulates 3,4 dimethylcatechol from o-xylene and both 3- and 4-methylcatechol from toluene. This means that there are two initial routes of oxidation of toluene by the strain. Pulsed-field gel electrophoresis analysis demonstrated the presence of two large megaplasmids in the wild-type strain DK17, one of which (pDK2) was lost in the mutant strain DK176. Since several other independently derived mutant strains unable to grow on alkylbenzenes are also missing pDK2, the genes encoding the initial steps in alkylbenzene metabolism (but not phenol metabolism) appear to be present on this approximately 330-kb plasmid. PMID- 12089005 TI - Process and metabolic strategies for improved production of Escherichia coli derived 6-deoxyerythronolide B. AB - Recently, the feasibility of using Escherichia coli for the heterologous biosynthesis of complex polyketides has been demonstrated. In this report, the development of a robust high-cell-density fed-batch procedure for the efficient production of complex polyketides is described. The effects of various physiological conditions on the productivity and titers of 6-deoxyerythronolide B (6dEB; the macrocyclic core of the antibiotic erythromycin) in recombinant cultures of E. coli were studied in shake flask cultures. The resulting data were used as a foundation to develop a high-cell-density fermentation procedure by building upon procedures reported earlier for recombinant protein production in E. coli. The fermentation strategy employed consistently produced approximately 100 mg of 6dEB per liter, whereas shake flask conditions generated between 1 and 10 mg per liter. The utility of an accessory thioesterase (TEII from Saccharopolyspora erythraea) for enhancing the productivity of 6dEB in E. coli was also demonstrated (increasing the final titer of 6dEB to 180 mg per liter). In addition to reinforcing the potential for using E. coli as a heterologous host for wild-type- and engineered-polyketide biosynthesis, the procedures described in this study may be useful for the production of secondary metabolites that are difficult to access by other routes. PMID- 12089004 TI - Cloning and heterologous expression of an enantioselective amidase from Rhodococcus erythropolis strain MP50. AB - The gene for an enantioselective amidase was cloned from Rhodococcus erythropolis MP50, which utilizes various aromatic nitriles via a nitrile hydratase/amidase system as nitrogen sources. The gene encoded a protein of 525 amino acids which corresponded to a protein with a molecular mass of 55.5 kDa. The deduced complete amino acid sequence showed homology to other enantioselective amidases from different bacterial genera. The nucleotide sequence approximately 2.5 kb upstream and downstream of the amidase gene was determined, but no indications for a structural coupling of the amidase gene with the genes for a nitrile hydratase were found. The amidase gene was carried by an approximately 40-kb circular plasmid in R. erythropolis MP50. The amidase was heterologously expressed in Escherichia coli and shown to hydrolyze 2-phenylpropionamide, alpha chlorophenylacetamide, and alpha-methoxyphenylacetamide with high enantioselectivity; mandeloamide and 2-methyl-3-phenylpropionamide were also converted, but only with reduced enantioselectivity. The recombinant E. coli strain which synthesized the amidase gene was shown to grow with organic amides as nitrogen sources. A comparison of the amidase activities observed with whole cells or cell extracts of the recombinant E. coli strain suggested that the transport of the amides into the cells becomes the rate-limiting step for amide hydrolysis in recombinant E. coli strains. PMID- 12089006 TI - Potential repair of Escherichia coli DNA following exposure to UV radiation from both medium- and low-pressure UV sources used in drinking water treatment. AB - The increased use of UV radiation as a drinking water treatment technology has instigated studies of the repair potential of microorganisms following treatment. This study challenged the repair potential of an optimally grown nonpathogenic laboratory strain of Escherichia coli after UV radiation from low- and medium pressure lamps. Samples were irradiated with doses of 5, 8, and 10 mJ/cm(2) from a low-pressure lamp and 3, 5, 8, and 10 mJ/cm(2) from a medium-pressure UV lamp housed in a bench-scale collimated beam apparatus. Following irradiation, samples were incubated at 37 degrees C under photoreactivating light or in the dark. Sample aliquots were analyzed for up to 4 h following incubation using a standard plate count. Results of this study showed that E. coli underwent photorepair following exposure to the low-pressure UV source, but no repair was detectable following exposure to the medium-pressure UV source at the initial doses examined. Minimal repair was eventually observed upon medium-pressure UV lamp exposure when doses were lowered to 3 mJ/cm(2). This study clearly indicates differences in repair potential under laboratory conditions between irradiation from low-pressure and medium-pressure UV sources of the type used in water treatment. PMID- 12089007 TI - Tightly bound binary toxin in the cell wall of Bacillus sphaericus. AB - We have shown that urea-extracted cell wall of entomopathogenic Bacillus sphaericus 2297 and some other strains is a potent larvicide against Culex pipiens mosquitoes, with 50% lethal concentrations comparable to that of the well known B. sphaericus binary toxin, with which it acts synergistically. The wall toxicity develops in B. sphaericus 2297 cultures during the late logarithmic stage, earlier than the appearance of the binary toxin crystal. It disappears with sporulation when the binary toxin activity reaches its peak. Disruption of the gene for the 42-kDa protein (P42) of the binary toxin abolishes both cell wall toxicity and crystal formation. However, the cell wall of B. sphaericus 2297, lacking P42, kills C. pipiens larvae when mixed with Escherichia coli cells expressing P42. Thus, the cell wall toxicity in strongly toxic B. sphaericus strains must be attributed to the presence in the cell wall of tightly bound 51 kDa (P51) and P42 binary toxin proteins. The synergism between binary toxin crystals and urea-treated cell wall preparations reflects suboptimal distribution of binary toxin subunits in both compartments. Binary toxin crystal is slightly deficient in P51, while cell wall is lacking in P42. PMID- 12089008 TI - Survival of GacS/GacA mutants of the biological control bacterium Pseudomonas aureofaciens 30-84 in the wheat rhizosphere. AB - GacS/GacA comprises a two-component regulatory system that controls the expression of secondary metabolites required for the control of plant diseases in many pseudomonads. High mutation frequencies of gacS and gacA have been observed in liquid culture. We examined whether gacS/gacA mutants could competitively displace the wild-type populations on roots and thus pose a threat to the efficacy of biological control. The survival of a gac mutant alone and in competition with the wild type on roots was examined in the biological control strain Pseudomonas aureofaciens 30-84. In this bacterium, GacS/GacA controls the expression of phenazine antibiotics that are inhibitory to plant pathogenic fungi and enhance the competitive survival of the bacterium. Wheat seedlings were inoculated with strain 30-84, and bacteria were recovered from roots after 21 days in sterile or nonsterile soil to check for the presence of gacS or gacA mutants. Although no mutants were detected in the inoculum, gacS/gacA mutants were recovered from 29 out of 31 roots and comprised up to 36% of the total bacterial populations. Southern hybridization analysis of the recovered gacA mutants did not indicate a conserved mutational mechanism. Replacement series analysis on roots utilizing strain 30-84 and a gacA mutant (30-84.gacA) or a gacS mutant (30-84.A2) demonstrated that although the mutant population partially displaced the wild type in sterile soil, it did not do so in natural soil. In fact, in natural soil final rhizosphere populations of wild-type strain 30-84 starting from mixtures were at least 1.5 times larger than would be predicted from their inoculation ratio and generally were greater than or equal to the population of wild type alone despite lower inoculation rates. These results indicate that although gacS/gacA mutants survive in natural rhizosphere populations, they do not displace wild-type populations. Better survival of wild type populations in mixtures with mutants suggests that mutants arising de novo or introduced within the inoculum may be beneficial for the survival of wild-type populations in the rhizosphere. PMID- 12089009 TI - Phage-displayed peptides as developmental agonists for Phytophthora capsici zoospores. AB - As part of its pathogenic life cycle, Phytophthora capsici disperses to plants through a motile zoospore stage. Molecules on the zoospore surface are involved in reception of environmental signals that direct preinfection behavior. We developed a phage display protocol to identify peptides that bind to the surface molecules of P. capsici zoospores in vitro. The selected phage-displayed peptides contained an abundance of polar amino acids and proline but were otherwise not conserved. About half of the selected phage that were tested concomitantly induced zoospore encystment in the absence of other signaling agents. A display phage was shown to bind to the zoospore but not to the cyst form of P. capsici. Two free peptides corresponding to active phage were similarly able to induce encystment of zoospores, indicating that their ability to serve as signaling ligands did not depend on their exact molecular context. Isolation and subsequent expression of peptides that act on pathogens could allow the identification of receptor molecules on the zoospore surface, in addition to forming the basis for a novel plant disease resistance strategy. PMID- 12089010 TI - Identification of glyA (encoding serine hydroxymethyltransferase) and its use together with the exporter ThrE to increase L-threonine accumulation by Corynebacterium glutamicum. AB - L-threonine can be made by the amino acid-producing bacterium Corynebacterium glutamicum. However, in the course of this process, some of the L-threonine is degraded to glycine. We detected an aldole cleavage activity of L-threonine in crude extracts with an activity of 2.2 nmol min(-1) (mg of protein)(-1). In order to discover the molecular reason for this activity, we cloned glyA, encoding serine hydroxymethyltransferase (SHMT). By using affinity-tagged glyA, SHMT was isolated and its substrate specificity was determined. The aldole cleavage activity of purified SHMT with L-threonine as the substrate was 1.3 micromol min( 1) (mg of protein)(-1), which was 4% of that with L-serine as substrate. Reduction of SHMT activity in vivo was obtained by placing the essential glyA gene in the chromosome under the control of P(tac), making glyA expression isopropylthiogalactopyranoside dependent. In this way, the SHMT activity in an L threonine producer was reduced to 8% of the initial activity, which led to a 41% reduction in glycine, while L-threonine was simultaneously increased by 49%. The intracellular availability of L-threonine to aldole cleavage was also reduced by overexpressing the L-threonine exporter thrE. In C. glutamicum DR-17, which overexpresses thrE, accumulation of 67 mM instead of 49 mM L-threonine was obtained. This shows that the potential for amino acid formation can be considerably improved by reducing its intracellular degradation and increasing its export. PMID- 12089011 TI - Plant-dependent genotypic and phenotypic diversity of antagonistic rhizobacteria isolated from different Verticillium host plants. AB - To study the effect of plant species on the abundance and diversity of bacterial antagonists, the abundance, the phenotypic diversity, and the genotypic diversity of rhizobacteria isolated from potato, oilseed rape, and strawberry and from bulk soil which showed antagonistic activity towards the soilborne pathogen Verticillium dahliae Kleb. were analyzed. Rhizosphere and soil samples were taken five times over two growing seasons in 1998 and 1999 from a randomized field trial. Bacterial isolates were obtained after plating on R2A (Difco, Detroit, Mich.) or enrichment in microtiter plates containing high-molecular-weight substrates followed by plating on R2A. A total of 5,854 bacteria isolated from the rhizosphere of strawberry, potato, or oilseed rape or bulk soil from fallow were screened by dual testing for in vitro antagonism towards VERTICILLIUM: The proportion of isolates with antagonistic activity was highest for strawberry rhizosphere (9.5%), followed by oilseed rape (6.3%), potato (3.7%), and soil (3.3%). The 331 Verticillium antagonists were identified by their fatty acid methyl ester profiles. They were characterized by testing their in vitro antagonism against other pathogenic fungi; their glucanolytic, chitinolytic, and proteolytic activities; and their BOX-PCR fingerprints. The abundance and composition of Verticillium antagonists was plant species dependent. A rather high proportion of antagonists from the strawberry rhizosphere was identified as Pseudomonas putida B (69%), while antagonists belonging to the Enterobacteriaceae (Serratia spp., Pantoea agglomerans) were mainly isolated from the rhizosphere of oilseed rape. For P. putida A and B plant-specific genotypes were observed, suggesting that these bacteria were specifically enriched in each rhizosphere. PMID- 12089012 TI - Wine yeast strains engineered for glycogen overproduction display enhanced viability under glucose deprivation conditions. AB - We used metabolic engineering to produce wine yeasts with enhanced resistance to glucose deprivation conditions. Glycogen metabolism was genetically modified to overproduce glycogen by increasing the glycogen synthase activity and eliminating glycogen phosphorylase activity. All of the modified strains had a higher glycogen content at the stationary phase, but accumulation was still regulated during growth. Strains lacking GPH1, which encodes glycogen phosphorylase, are unable to mobilize glycogen. Enhanced viability under glucose deprivation conditions occurs when glycogen accumulates in the strain that overexpresses GSY2, which encodes glycogen synthase and maintains normal glycogen phosphorylase activity. This enhanced viability is observed under laboratory growth conditions and under vinification conditions in synthetic and natural musts. Wines obtained from this modified strain and from the parental wild-type strain don't differ significantly in the analyzed enological parameters. The engineered strain might better resist some stages of nutrient depletion during industrial use. PMID- 12089013 TI - In situ transfer of antibiotic resistance genes from transgenic (transplastomic) tobacco plants to bacteria. AB - Interkingdom gene transfer is limited by a combination of physical, biological, and genetic barriers. The results of greenhouse experiments involving transplastomic plants (genetically engineered chloroplast genomes) cocolonized by pathogenic and opportunistic soil bacteria demonstrated that these barriers could be eliminated. The Acinetobacter sp. strain BD413, which is outfitted with homologous sequences to chloroplastic genes, coinfected a transplastomic tobacco plant with Ralstonia solanacearum and was transformed by the plant's transgene (aadA) containing resistance to spectinomycin and streptomycin. However, no transformants were observed when the homologous sequences were omitted from the Acinetobacter sp. strain. Detectable gene transfer from these transgenic plants to bacteria were dependent on gene copy number, bacterial competence, and the presence of homologous sequences. Our data suggest that by selecting plant transgene sequences that are nonhomologous to bacterial sequences, plant biotechnologists could restore the genetic barrier to transgene transfer to bacteria. PMID- 12089014 TI - Molecular characterization and expression of pyruvate formate-lyase-activating enzyme in a ruminal bacterium, Streptococcus bovis. AB - To clarify the significance of the activation of pyruvate formate-lyase (PFL) by PFL-activating enzyme (PFL-AE) in Streptococcus bovis, the molecular properties and gene expression of PFL-AE were investigated. S. bovis PFL-AE was deduced to consist of 261 amino acids with a molecular mass of 29.9 kDa and appeared to be a monomer protein. Similar to Escherichia coli PFL-AE, S. bovis PFL-AE required Fe(2+) for activity. The gene encoding PFL-AE (act) was found to be polycistronic, and the PFL gene (pfl) was not included. However, the act mRNA level changed in parallel with the pfl mRNA level, responding to growth conditions, and the change was contrary to the change in the lactate dehydrogenase (LDH) mRNA level. PFL-AE synthesis appeared to change in parallel with PFL synthesis. Introduction of a recombinant plasmid containing S. bovis pfl and the pfl promoter into S. bovis did not affect formate and lactate production, which suggests that the activity of the pfl promoter is low. When the pfl promoter was replaced by the S. bovis ldh promoter, PFL was overexpressed, which caused an increase in the formate-to-lactate ratio. However, when PFL-AE was overexpressed, the formate-to-lactate ratio did not change, suggesting that PFL AE was present at a level that was high enough to activate PFL. When both PFL-AE and PFL were overexpressed, the formate-to-lactate ratio further increased. It is conceivable that LDH activity is much higher than PFL activity, which may explain why the formate-to-lactate ratio is usually low. PMID- 12089015 TI - Seasonal fluctuations and long-term persistence of pathogenic populations of Agrobacterium spp. in soils. AB - Short- and long-term persistence of pathogenic (i.e., tumor forming) agrobacteria in soil was investigated in six nursery plots with a history of high crown gall incidence. No pathogenic Agrobacterium strains were isolated in soil samples taken in fall and winter in any plots, but such strains were isolated from both bulk soils and weed rhizospheres (over 0.5 x 10(5) pathogenic CFU/g of bulk soil or rhizosphere) in three out of six plots in spring and summer. PCR amplifications of a vir sequence from DNA extracted from soil confirmed the presence of Ti plasmids in summer and their absence in fall and winter. The results indicate that strains that harbor a Ti plasmid had an unforeseen positive fitness versus Ti plasmid-free strains in soil and rhizosphere in spring and summer in spite of the apparent absence of tumor, and hence of opines. The gain of fitness occurred during a bloom of all cultivable agrobacteria observed only in conducive soils. An evolution of the pathogenic population was recorded during a 4-year period in one particularly conducive soil. In 1990, the pathogenic population in this soil consisted of only biovar 1 strains harboring both octopine- and nopaline-type Ti plasmids. In 1994, it consisted of only nopaline type Ti plasmids equally distributed among biovar 1 and 2 strains. These results suggest that nopaline-type Ti plasmids conferred a better survival ability than octopine-type Ti plasmids to biovar 2 agrobacteria under the present field conditions. PMID- 12089016 TI - Prevalence and fingerprinting of Listeria monocytogenes strains isolated from raw whole milk in farm bulk tanks and in dairy plant receiving tanks. AB - The incidence of Listeria species in raw whole milk from farm bulk tanks and from raw milk in storage at a Swedish dairy plant was studied. Listeria monocytogenes was found in 1.0% and Listeria innocua was found in 2.3% of the 294 farm bulk tank (farm tank) milk specimens. One farm tank specimen contained 60 CFU of L. monocytogenes ml(-1). L. monocytogenes was detected in 19.6% and L. innocua was detected in 8.5% of the milk specimens from the silo receiving tanks at the dairy (dairy silos). More dairy silo specimens were positive for both Listeria species during winter than during summer. Restriction enzyme analysis and pulsed-field gel electrophoresis were applied to 65 isolates of L. monocytogenes, resulting in 16 different clonal types. Two clonal types were shared by the farm tank milk and the dairy silo milk. All except one clonal type belonged to serovar 1/2a. In the dairy silo milk five clonal types were found more frequently and for a longer period than the others. No Listeria species were found in any other samples from the plant. PMID- 12089017 TI - Identification of an opd (organophosphate degradation) gene in an Agrobacterium isolate. AB - We isolated a bacterial strain, Agrobacterium radiobacter P230, which can hydrolyze a wide range of organophosphate (OP) insecticides. A gene encoding a protein involved in OP hydrolysis was cloned from A. radiobacter P230 and sequenced. This gene (called opdA) had sequence similarity to opd, a gene previously shown to encode an OP-hydrolyzing enzyme in Flavobacterium sp. strain ATCC 27551 and Brevundimonas diminuta MG. Insertional mutation of the opdA gene produced a strain lacking the ability to hydrolyze OPs, suggesting that this is the only gene encoding an OP-hydrolyzing enzyme in A. radiobacter P230. The OPH and OpdA proteins, encoded by opd and opdA, respectively, were overexpressed and purified as maltose-binding proteins, and the maltose-binding protein moiety was cleaved and removed. Neither protein was able to hydrolyze the aliphatic OP malathion. The kinetics of the two proteins for diethyl OPs were comparable. For dimethyl OPs, OpdA had a higher k(cat) than OPH. It was also capable of hydrolyzing the dimethyl OPs phosmet and fenthion, which were not hydrolyzed at detectable levels by OPH. PMID- 12089018 TI - Technical-scale production of cyanophycin with recombinant strains of Escherichia coli. AB - By the use of Escherichia coli DH1 harboring cphA from Synechocystis sp. strain PCC6803, large-scale production of cyanophycin at 30- and 500-liter culture volumes was established. Transcription of cphA was controlled by the thermosensitive cI857 repressor, which enabled induction of cphA by a simple temperature shift in the culture fluid. Maximum cyanophycin cell content of up to 24% (wt/wt) of cellular dry matter was obtained by induction in the early exponential growth phase and cultivation of the cells in terrific broth complex medium. Synthesis of cyanophycin was found to be strongly dependent on the presence of complex components, and in mineral salts medium the cells synthesized and accumulated cyanophycin only if Casamino Acids were added. Cultivations were done at the 500-liter scale, allowing the provision of cell mass for the preparation of cyanophycin at the kilogram scale. Isolation of cyanophycin was achieved by a new acid extraction procedure which allowed large-scale purification of the polyamide from whole cells. PMID- 12089019 TI - Expression of a fungal hydrophobin in the Saccharomyces cerevisiae cell wall: effect on cell surface properties and immobilization. AB - The aim of this work was to modify the cell surface properties of Saccharomyces cerevisiae by expression of the HFBI hydrophobin of the filamentous fungus Trichoderma reesei on the yeast cell surface. The second aim was to study the immobilization capacity of the modified cells. Fusion to the Flo1p flocculin was used to target the HFBI moiety to the cell wall. Determination of cell surface characteristics with contact angle and zeta potential measurements indicated that HFBI-producing cells are more apolar and slightly less negatively charged than the parent cells. Adsorption of the yeast cells to different commercial supports was studied. A twofold increase in the binding affinity of the hydrophobin producing yeast to hydrophobic silicone-based materials was observed, while no improvement in the interaction with hydrophilic carriers could be seen compared to that of the parent cells. Hydrophobic interactions between the yeast cells and the support are suggested to play a major role in attachment. Also, a slight increase in the initial adsorption rate of the hydrophobin yeast was observed. Furthermore, due to the engineered cell surface, hydrophobin-producing yeast cells were efficiently separated in an aqueous two-phase system by using a nonionic polyoxyethylene detergent, C(12-18)EO(5). PMID- 12089020 TI - Heterologous expression of the Lactococcus lactis bacteriocin, nisin, in a dairy Enterococcus strain. AB - The bacteriocin nisin is produced only by some strains of Lactococcus lactis, and to date production in other lactic acid bacteria has not been achieved. Enterococcus sp. strain N12beta is a nisin-immune transconjugant obtained from a nisin-producing donor (L. lactis ATCC 11454) and a dairy recipient (Enterococcus sp. strain S12beta), but it does not produce nisin. In this study, using PCR amplification, we confirmed that the whole nisin operon is likely present in Enterococcus sp. strain N12beta. Northern hybridization of total RNA from strain N12beta with a nisA probe and the results of reverse transcriptase PCR showed the lack of nisA transcription in this strain. However, nisA transcription was partially restored in strain N12beta upon growth in the presence of exogenous nisin, and the nisA transcription signal was intensified after an increase in the external nisin level. Furthermore, bioassays showed that active nisin was produced in a dose-dependent fashion by strain N12beta following induction by exogenous nisin. These results indicated that expression of the nisin genes in Enterococcus sp. strain N12beta depended on autoinduction via signal transduction. However, the amount of external inducing signal required was significantly greater than the amount needed for autoinduction in L. lactis. PMID- 12089021 TI - Mucosa-associated bacteria in the human gastrointestinal tract are uniformly distributed along the colon and differ from the community recovered from feces. AB - The human gastrointestinal (GI) tract harbors a complex community of bacterial cells in the mucosa, lumen, and feces. Since most attention has been focused on bacteria present in feces, knowledge about the mucosa-associated bacterial communities in different parts of the colon is limited. In this study, the bacterial communities in feces and biopsy samples from the ascending, transverse, and descending colons of 10 individuals were analyzed by using a 16S rRNA approach. Flow cytometric analysis indicated that 10(5) to 10(6) bacteria were present in the biopsy samples. To visualize the diversity of the predominant and the Lactobacillus group community, denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene amplicons was performed. DGGE analysis and similarity index comparisons demonstrated that the predominant mucosa-associated bacterial community was host specific and uniformly distributed along the colon but significantly different from the fecal community (P < 0.01). The Lactobacillus group-specific profiles were less complex than the profiles reflecting the predominant community. For 6 of the 10 individuals the community of Lactobacillus like bacteria in the biopsy samples was similar to that in the feces. Amplicons having 99% sequence similarity to the 16S ribosomal DNA of Lactobacillus gasseri were detected in the biopsy samples of nine individuals. No significant differences were observed between healthy and diseased individuals. The observed host-specific DGGE profiles of the mucosa-associated bacterial community in the colon support the hypothesis that host-related factors are involved in the determination of the GI tract microbial community. PMID- 12089022 TI - Improving the pathogenicity of a nematode-trapping fungus by genetic engineering of a subtilisin with nematotoxic activity. AB - Nematophagous fungi are soil-living fungi that are used as biological control agents of plant and animal parasitic nematodes. Their potential could be improved by genetic engineering, but the lack of information about the molecular background of the infection has precluded this development. In this paper we report that a subtilisin-like extracellular serine protease designated PII is an important pathogenicity factor in the common nematode-trapping fungus Arthrobotrys oligospora. The transcript of PII was not detected during the early stages of infection (adhesion and penetration), but high levels were expressed concurrent with the killing and colonization of the nematode. Disruption of the PII gene by homologous recombination had a limited effect on the pathogenicity of the fungus. However, mutants containing additional copies of the PII gene developed a higher number of infection structures and had an increased speed of capturing and killing nematodes compared to the wild type. The paralyzing activity of PII was verified by demonstrating that a heterologous-produced PII (in Aspergillus niger) had a nematotoxic activity when added to free-living nematodes. The toxic activity of PII was significantly higher than that of other commercially available serine proteases. This is the first report showing that genetic engineering can be used to improve the pathogenicity of a nematode trapping fungus. In the future it should be possible to express recombinant subtilisins with nematicidal activity in other organisms that are present in the habitat of parasitic nematodes (e.g., host plant). PMID- 12089023 TI - Antibiotic and biosurfactant properties of cyclic lipopeptides produced by fluorescent Pseudomonas spp. from the sugar beet rhizosphere. AB - Cyclic lipopeptides (CLPs) with antibiotic and biosurfactant properties are produced by a number of soil bacteria, including fluorescent Pseudomonas spp. To provide new and efficient strains for the biological control of root-pathogenic fungi in agricultural crops, we isolated approximately 600 fluorescent Pseudomonas spp. from two different agricultural soils by using three different growth media. CLP production was observed in a large proportion of the strains (approximately 60%) inhabiting the sandy soil, compared to a low proportion (approximately 6%) in the loamy soil. Chemical structure analysis revealed that all CLPs could be clustered into two major groups, each consisting of four subgroups. The two major groups varied primarily in the number of amino acids in the cyclic peptide moiety, while each of the subgroups could be differentiated by substitutions of specific amino acids in the peptide moiety. Production of specific CLPs could be affiliated with Pseudomonas fluorescens strain groups belonging to biotype I, V, or VI. In vitro analysis using both purified CLPs and whole-cell P. fluorescens preparations demonstrated that all CLPs exhibited strong biosurfactant properties and that some also had antibiotic properties towards root-pathogenic microfungi. The CLP-producing P. fluorescens strains provide a useful resource for selection of biological control agents, whether a single strain or a consortium of strains was used to maximize the synergistic effect of multiple antagonistic traits in the inoculum. PMID- 12089024 TI - Distribution of genes encoding the trypsin-dependent lantibiotic ruminococcin A among bacteria isolated from human fecal microbiota. AB - Fourteen bacterial strains capable of producing a trypsin-dependent antimicrobial substance active against Clostridium perfringens were isolated from human fecal samples of various origins (from healthy adults and children, as well as from adults with chronic pouchitis). Identification of these strains showed that they belonged to Ruminococcus gnavus, Clostridium nexile, and Ruminococcus hansenii species or to new operational taxonomic units, all from the Clostridium coccoides phylogenetic group. In hybridization experiments with a probe specific for the structural gene encoding the trypsin-dependent lantibiotic ruminococcin A (RumA) produced by R. gnavus, seven strains gave a positive response. All of them harbored three highly conserved copies of rumA-like genes. The deduced peptide sequence was identical to or showed one amino acid difference from the hypothetical precursor of RumA. Our results indicate that the rumA-like genes have been disseminated among R. gnavus and phylogenetically related strains that can make up a significant part of the human fecal microbiota. PMID- 12089025 TI - Modeling the interactions of Lactobacillus curvatus colonies in solid medium: consequences for food quality and safety. AB - The growth process of Lactobacillus curvatus colonies was quantified by a coupled growth and diffusion equation incorporating a volumetric rate of lactic acid production. Analytical solutions were compared to numerical ones, and both were able to predict the onset of interaction well. The derived analytical solution modeled the lactic acid concentration profile as a function of the diffusion coefficient, colony radius, and volumetric production rate. Interaction was assumed to occur when the volume-averaged specific growth rate of the cells in a colony was 90% of the initial maximum rate. Growth of L. curvatus in solid medium is dependent on the number of cells in a colony. In colonies with populations of fewer than 10(5) cells, mass transfer limitation is not significant for the growth process. When the initial inoculation density is relatively high, colonies are not able to grow to these sizes and growth approaches that of broth cultures (negligible mass transfer limitation). In foods, which resemble the model solid system and in which the initial inoculation density is high, it will be appropriate to use predictive models of broth cultures to estimate growth. For a very low initial inoculation density, large colonies can develop that will start to deviate from growth in broth cultures, but only after large outgrowth. PMID- 12089026 TI - Degradation of humic acids by the litter-decomposing basidiomycete Collybia dryophila. AB - The basidiomycete Collybia dryophila K209, which colonizes forest soil, was found to decompose a natural humic acid isolated from pine-forest litter (LHA) and a synthetic (14)C-labeled humic acid ((14)C-HA) prepared from [U-(14)C]catechol in liquid culture. Degradation resulted in the formation of polar, lower-molecular mass fulvic acid (FA) and carbon dioxide. HA decomposition was considerably enhanced in the presence of Mn(2+) (200 microM), leading to 75% conversion of LHA and 50% mineralization of (14)C-HA (compared to 60% and 20%, respectively, in the absence of Mn(2+)). There was a strong indication that manganese peroxidase (MnP), the production of which was noticeably increased in Mn(2+)-supplemented cultures, was responsible for this effect. The enzyme was produced as a single protein with a pI of 4.7 and a molecular mass of 44 kDa. During solid-state cultivation, C. dryophila released substantial amounts of water-soluble FA (predominantly of 0.9 kDa molecular mass) from insoluble litter material. The results indicate that basidiomycetes such as C. dryophila which colonize forest litter and soil are involved in humus turnover by their recycling of high molecular-mass humic substances. Extracellular MnP seems to be a key enzyme in the conversion process. PMID- 12089027 TI - tfdA-like genes in 2,4-dichlorophenoxyacetic acid-degrading bacteria belonging to the Bradyrhizobium-Agromonas-Nitrobacter-Afipia cluster in alpha-Proteobacteria. AB - The 2,4-dichlorophenoxyacetate (2,4-D)/alpha-ketoglutarate dioxygenase gene (tfdA) homolog designated tfdAalpha was cloned and characterized from 2,4-D degrading bacterial strain RD5-C2. This Japanese upland soil isolate belongs to the Bradyrhizobium-Agromonas-Nitrobacter-Afipia cluster in the alpha subdivision of the class Proteobacteria on the basis of its 16S ribosomal DNA sequence. Sequence analysis showed 56 to 60% identity of tfdAalpha to representative tfdA genes. A MalE-TfdAalpha fusion protein expressed in Escherichia coli exhibited about 10 times greater activity for phenoxyacetate than 2,4-D in an alpha ketoglutarate- and Fe(II)-dependent reaction. The deduced amino acid sequence of TfdAalpha revealed a conserved His-X-Asp-X(146)-His-X(14)-Arg motif characteristic of the active site of group II alpha-ketoglutarate-dependent dioxygenases. The tfdAalpha genes were also detected in 2,4-D-degrading alpha Proteobacteria previously isolated from pristine environments in Hawaii and in Saskatchewan, Canada (Y. Kamagata, R. R. Fulthorpe, K. Tamura, H. Takami, L. J. Forney, and J. M. Tiedje, Appl. Environ. Microbiol. 63:2266-2272, 1997). These findings indicate that the tfdA genes in beta- and gamma-Proteobacteria and the tfdAalpha genes in alpha-Proteobacteria arose by divergent evolution from a common ancestor. PMID- 12089028 TI - Bovine enteroviruses as indicators of fecal contamination. AB - Surface waters frequently have been contaminated with human enteric viruses, and it is likely that animal enteric viruses have contaminated surface waters also. Bovine enteroviruses (BEV), found in cattle worldwide, usually cause asymptomatic infections and are excreted in the feces of infected animals in large numbers. In this study, the prevalence and genotype of BEV in a closed herd of cattle were evaluated and compared with BEV found in animals in the immediate environment and in environmental specimens. BEV was found in feces from 76% of cattle, 38% of white-tailed deer, and one of three Canada geese sharing the same pastures, as well as the water obtained from animal watering tanks, from the pasture, from streams running from the pasture to an adjacent river, and from the river, which emptied into the Chesapeake Bay. Furthermore, BEV was found in oysters collected from that river downstream from the farm. These findings suggest that BEV could be used as an indicator of fecal pollution originating from animals (cattle and/or deer). Partial sequence analysis of the viral genomes indicates that different viral variants coexist in the same area. The possibility of identifying the viral strains found in the animals and in the contaminated areas by sequencing the RNA genome, could provide a tool to find the origin of the contamination and should be useful for epidemiological and viral molecular evolution studies. PMID- 12089029 TI - Klebsiella pneumoniae produces no histamine: Raoultella planticola and Raoultella ornithinolytica strains are histamine producers. AB - Histamine fish poisoning is caused by histamine-producing bacteria (HPB). Klebsiella pneumoniae and Klebsiella oxytoca are the best-known HPB in fish. However, 22 strains of HPB from fish first identified as K. pneumoniae or K. oxytoca by commercialized systems were later correctly identified as Raoultella planticola (formerly Klebsiella planticola) by additional tests. Similarly, five strains of Raoultella ornithinolytica (formerly Klebsiella ornithinolytica) were isolated from fish as new HPB. R. planticola and R. ornithinolytica strains were equal in their histamine-producing capabilities and were determined to possess the hdc genes, encoding histidine decarboxylase. On the other hand, a collection of 61 strains of K. pneumoniae and 18 strains of K. oxytoca produced no histamine. PMID- 12089030 TI - Genotypic and phenotypic diversity within species of purple nonsulfur bacteria isolated from aquatic sediments. AB - To assess the extent of genotypic and phenotypic diversity within species of purple nonsulfur bacteria found in aquatic sediments, a total of 128 strains were directly isolated from agar plates that had been inoculated with sediment samples from Haren and De Biesbosch in The Netherlands. All isolates were initially characterized by BOX-PCR genomic DNA fingerprinting, and 60 distinct genotypes were identified. Analyses of 16S rRNA gene sequences of representatives of each genotype showed that five and eight different phylotypes of purple nonsulfur bacteria were obtained from the Haren and De Biesbosch sites, respectively. At the Haren site, 80.5% of the clones were Rhodopseudomonas palustris, whereas Rhodoferax fermentans and Rhodopseudomonas palustris were numerically dominant at the De Biesbosch site and constituted 45.9 and 34.4% of the isolates obtained, respectively. BOX-PCR genomic fingerprints showed that there was a high level of genotypic diversity within each of these species. The genomic fingerprints of Rhodopseudomonas palustris isolates were significantly different for isolates from the two sampling sites, suggesting that certain strains may be endemic to each sampling site. Not all Rhodopseudomonas palustris isolates could degrade benzoate, a feature that has previously been thought to be characteristic of the species. There were differences in the BOX-PCR genomic fingerprints and restriction fragment length polymorphisms of benzoate-coenzyme A ligase genes and form I and form II ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) genes between benzoate-degrading and non-benzoate-degrading genotypes. The ability to distinguish these two Rhodopseudomonas palustris groups based on multiple genetic differences may reflect an incipient speciation event resulting from adaptive evolution to local environmental conditions. PMID- 12089031 TI - Growth in coculture stimulates metabolism of the phenylurea herbicide isoproturon by Sphingomonas sp. strain SRS2. AB - Metabolism of the phenylurea herbicide isoproturon by Sphingomonas sp. strain SRS2 was significantly enhanced when the strain was grown in coculture with a soil bacterium (designated strain SRS1). Both members of this consortium were isolated from a highly enriched isoproturon-degrading culture derived from an agricultural soil previously treated regularly with the herbicide. Based on analysis of the 16S rRNA gene, strain SRS1 was assigned to the beta-subdivision of the proteobacteria and probably represents a new genus. Strain SRS1 was unable to degrade either isoproturon or its known metabolites 3-(4-isopropylphenyl)-1 methylurea, 3-(4-isopropylphenyl)-urea, or 4-isopropyl-aniline. Pure culture studies indicate that Sphingomonas sp. SRS2 is auxotrophic and requires components supplied by association with other soil bacteria. A specific mixture of amino acids appeared to meet these requirements, and it was shown that methionine was essential for Sphingomonas sp. SRS2. This suggests that strain SRS1 supplies amino acids to Sphingomonas sp. SRS2, thereby leading to rapid metabolism of (14)C-labeled isoproturon to (14)CO(2) and corresponding growth of strain SRS2. Proliferation of strain SRS1 suggests that isoproturon metabolism by Sphingomonas sp. SRS2 provides unknown metabolites or cell debris that supports growth of strain SRS1. The role of strain SRS1 in the consortium was not ubiquitous among soil bacteria; however, the indigenous soil microflora and some strains from culture collections also stimulate isoproturon metabolism by Sphingomonas sp. strain SRS2 to a similar extent. PMID- 12089032 TI - Identification of proteins involved in the heat stress response of Bacillus cereus ATCC 14579. AB - To monitor the ability of the food-borne opportunistic pathogen Bacillus cereus to survive during minimal processing of food products, we determined its heat adaptive response. During pre-exposure to 42 degrees C, B. cereus ATCC 14579 adapts to heat exposure at the lethal temperature of 50 degrees C (maximum protection occurs after 15 min to 1 h of pre-exposure to 42 degrees C). For this heat-adaptive response, de novo protein synthesis is required. By using two dimensional gel electrophoresis, we observed 31 heat-induced proteins, and we determined the N-terminal sequences of a subset of these proteins. This revealed induction of stress proteins (CspB, CspE, and SodA), proteins involved in sporulation (SpoVG and AldA), metabolic enzymes (FolD and Dra), identified heat induced proteins in related organisms (DnaK, GroEL, ClpP, RsbV, HSP16.4, YflT, PpiB, and TrxA), and other proteins (MreB, YloH, and YbbT). The upregulation of several stress proteins was confirmed by using antibodies specific for well characterized heat shock proteins (HSPs) of B. subtilis. These observations indicate that heat adaptation of B. cereus involves proteins that function in a variety of cellular processes. Notably, a 30-min pre-exposure to 4% ethanol, pH 5, or 2.5% NaCl also results in increased thermotolerance. Also, for these adaptation processes, protein synthesis is required, and indeed, some HSPs are induced under these conditions. Collectively, these data show that during mild processing, cross-protection from heating occurs in pathogenic B. cereus, which may result in increased survival in foods. PMID- 12089033 TI - Influence of the transposition of the thermostabilizing domain of Clostridium thermocellum xylanase (XynX) on xylan binding and thermostabilization. AB - A xylanase gene, xynX, of Clostridium thermocellum had one thermostabilizing domain (TSD) between the signal peptide sequence and the catalytic domain (CD). The TSD of a truncated xylanase gene, xynX'(TSD-CD), was transpositioned from the N terminus to the C terminus of the CD by overlapping PCRs, and a modified product, xynX'(CD-TSD), was constructed. XynX'(TSD-CD) had a higher optimum temperature (70 degrees C versus 65 degrees C) and was more thermostable (residual activity of 68% versus 46% after a 20-min preincubation at 70 degrees C) than the one without the TSD, XynX'(CD). However, the domain-transpositioned enzyme, XynX'(CD-TSD), showed a lower optimum temperature (30 degrees C) and thermostability (20%) than XynX'(CD). Both XynX'(TSD-CD) and XynX'(CD-TSD) showed significantly higher binding capacity toward xylan than XynX'(CD), and the domain transposition did not cause any change in the binding ability. XynX'(TSD-CD) and XynX'(CD-TSD) also showed considerable binding to lichenan but not to carboxymethyl cellulose and laminarin. XynX'(TSD-CD) and XynX'(CD-TSD) had higher activities for insoluble xylan than XynX'(CD), while XynX'(CD) was more active against soluble xylan than XynX'(TSD-CD) and XynX'(CD-TSD). These results indicate that the TSD of XynX has dual functions, xylan binding and thermostabilization, and the domain should also be classified as a xylan-binding domain (XBD). The binding capacity of the XBD was not affected by domain transpositioning within the gene. PMID- 12089034 TI - Solid-phase contact assay that uses a lux-marked Nitrosomonas europaea reporter strain to estimate toxicity of bioavailable linear alkylbenzene sulfonate in soil. AB - Information about in situ toxicity of the bioavailable pools of adsorptive soil pollutants is a prerequisite for proper ecological risk assessment in contaminated soils. Such toxicity data may be obtained by assays allowing for direct exposure of introduced test microorganisms to the toxicants, as they appear in solid solution equilibria in the natural soil. We describe a novel sensitive solid-phase contact assay for in situ toxicity testing of soil pollutants based on a recombinant bioluminescent reporter strain of Nitrosomonas europaea. A slurry of the reporter strain and soil sample was shaken for 1 h, after which bioluminescence was measured either directly (soil slurry protocol) or in the supernatant obtained after centrifugation (soil extract protocol). The assay was validated for both protocols by using linear alkylbenzene sulfonate (LAS) as a toxic and adsorptive model compound in the soil samples. Interestingly, LAS showed the same toxicity to the reporter strain with either soil incubation (both protocols) or pure culture, suggesting that adsorbed LAS pools contributed to the observed toxicity. The solid-phase contact assay that used the reporter strain of lux-marked N. europaea was slightly more sensitive for the detection of LAS toxicity in soil than activity-based assays targeting indigenous nitrifiers and much more sensitive than assays targeting indigenous heterotrophic microbes. We conclude that the new solid-phase contact assay, which is based on direct interaction of the test microorganisms with bioavailable pools of the toxicants in soil, provides a most sensitive and relevant method for evaluating the in situ toxicity and assessing the risks of soil contaminants. PMID- 12089035 TI - Large fraction of dead and inactive bacteria in coastal marine sediments: comparison of protocols for determination and ecological significance. AB - It is now universally recognized that only a portion of aquatic bacteria is actively growing, but quantitative information on the fraction of living versus dormant or dead bacteria in marine sediments is completely lacking. We compared different protocols for the determination of the dead, dormant, and active bacterial fractions in two different marine sediments and at different depths into the sediment core. Bacterial counts ranged between (1.5 +/- 0.2) x 10(8) cells g(-1) and (53.1 +/- 16.0) x 10(8) cells g(-1) in sandy and muddy sediments, respectively. Bacteria displaying intact membrane (live bacterial cells) accounted for 26 to 30% of total bacterial counts, while dead cells represented the most abundant fraction (70 to 74%). Among living bacterial cells, nucleoid containing cells represented only 4% of total bacterial counts, indicating that only a very limited fraction of bacterial assemblage was actively growing. Nucleoid-containing cells increased with increasing sediment organic content. The number of bacteria responsive to antibiotic treatment (direct viable count; range, 0.3 to 4.8% of the total bacterial number) was significantly lower than nucleoid-containing cell counts. An experiment of nutrient enrichment to stimulate a response of the dormant bacterial fraction determined a significant increase of nucleoid-containing cells. After nutrient enrichment, a large fraction of dormant bacteria (6 to 11% of the total bacterial number) was "reactivated." Bacterial turnover rates estimated ranged from 0.01 to 0.1 day(-1) but were 50 to 80 times higher when only the fraction of active bacteria was considered (on average 3.2 day(-1)). Our results suggest that the fraction of active bacteria in marine sediments is controlled by nutrient supply and availability and that their turnover rates are at least 1 order of magnitude higher than previously reported. PMID- 12089036 TI - Laccase-catalyzed oxidation of Mn(2+) in the presence of natural Mn(3+) chelators as a novel source of extracellular H(2)O(2) production and its impact on manganese peroxidase. AB - A purified and electrophoretically homogeneous blue laccase from the litter decaying basidiomycete Stropharia rugosoannulata with a molecular mass of approximately 66 kDa oxidized Mn(2+) to Mn(3+), as assessed in the presence of the Mn chelators oxalate, malonate, and pyrophosphate. At rate-saturating concentrations (100 mM) of these chelators and at pH 5.0, Mn(3+) complexes were produced at 0.15, 0.05, and 0.10 micromol/min/mg of protein, respectively. Concomitantly, application of oxalate and malonate, but not pyrophosphate, led to H(2)O(2) formation and tetranitromethane (TNM) reduction indicative for the presence of superoxide anion radical. Employing oxalate, H(2)O(2) production, and TNM reduction significantly exceeded those found for malonate. Evidence is provided that, in the presence of oxalate or malonate, laccase reactions involve enzyme-catalyzed Mn(2+) oxidation and abiotic decomposition of these organic chelators by the resulting Mn(3+), which leads to formation of superoxide and its subsequent reduction to H(2)O(2). A partially purified manganese peroxidase (MnP) from the same organism did not produce Mn(3+) complexes in assays containing 1 mM Mn(2+) and 100 mM oxalate or malonate, but omitting an additional H(2)O(2) source. However, addition of laccase initiated MnP reactions. The results are in support of a physiological role of laccase-catalyzed Mn(2+) oxidation in providing H(2)O(2) for extracellular oxidation reactions and demonstrate a novel type of laccase-MnP cooperation relevant to biodegradation of lignin and xenobiotics. PMID- 12089038 TI - Purification and characterization of an extracellular protease from Penicillium chrysogenum Pg222 active against meat proteins. AB - An extracellular protease from Penicillium chrysogenum (Pg222) isolated from dry cured ham has been purified. The purification procedure involved several steps: ammonium sulfate precipitation, ion-exchange chromatography, filtration, and separation by high-performance liquid chromatography. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and gel filtration, the purified fraction showed a molecular mass of about 35 kDa. The hydrolytic properties of the purified enzyme (EPg222) on extracted pork myofibrillar proteins under several conditions were evaluated by SDS-PAGE. EPg222 showed activity in the range of 10 to 60 degrees C in temperature, 0 to 3 M NaCl, and pH 5 to 7, with maximum activity at pH 6, 45 degrees C, and 0.25 M NaCl. Under these conditions the enzyme was most active against tropomyosin, actin, and myosin. EPg222 showed collagenolytic activity but did not hydrolyze myoglobin. EPg222 showed higher activity than other proteolytic enzymes like papain, trypsin, and Aspergillus oryzae protease. The N-terminal amino acid sequence was determined and was found to be Glu-Asn-Pro-Leu-Gln-Pro-Asn-Ala-Pro-Ser-Trp. This partial amino acid sequence revealed a 55% homology with serine proteases from Penicillium citrinum. The activity of this novel protease may be of interest in ripening and generating the flavor of dry-cured meat products. PMID- 12089037 TI - Fungal fragments as indoor air biocontaminants. AB - The aerosolization process of fungal propagules of three species (Aspergillus versicolor, Penicillium melinii, and Cladosporium cladosporioides) was studied by using a newly designed and constructed aerosolization chamber. We discovered that fungal fragments are aerosolized simultaneously with spores from contaminated agar and ceiling tile surfaces. Concentration measurements with an optical particle counter showed that the fragments are released in higher numbers (up to 320 times) than the spores. The release of fungal propagules varied depending on the fungal species, the air velocity above the contaminated surface, and the texture and vibration of the contaminated material. In contrast to spores, the release of fragments from smooth surfaces was not affected by air velocity, indicating a different release mechanism. Correlation analysis showed that the number of released fragments cannot be predicted on the basis of the number of spores. Enzyme-linked immunosorbent assays with monoclonal antibodies produced against Aspergillus and Penicillium fungal species showed that fragments and spores share common antigens, which not only confirmed the fungal origin of the fragments but also established their potential biological relevance. The considerable immunological reactivity, the high number, and the small particle size of the fungal fragments may contribute to human health effects that have been detected in buildings with mold problems but had no scientific explanation until now. This study suggests that future fungal spore investigations in buildings with mold problems should include the quantitation of fungal fragments. PMID- 12089039 TI - Identification of the Enterococcus faecalis tyrosine decarboxylase operon involved in tyramine production. AB - Screening of a library of Enterococcus faecalis insertional mutants allowed isolation of a mutant affected in tyramine production. The growth of this mutant was similar to that of the wild-type E. faecalis JH2-2 strain in Maijala broth, whereas high-performance liquid chromatography analyses showed that tyramine production, which reached 1,000 microg ml(-1) for the wild-type strain, was completely abolished. Genetic analysis of the insertion locus revealed a gene encoding a decarboxylase with similarity to eukaryotic tyrosine decarboxylases. Sequence analysis revealed a pyridoxal phosphate binding site, indicating that this enzyme belongs to the family of amino acid decarboxylases using this cofactor. Reverse transcription-PCR analyses demonstrated that the gene (tdc) encoding the putative tyrosine decarboxylase of E. faecalis JH2-2 is cotranscribed with the downstream gene encoding a putative tyrosine-tyramine antiporter and with the upstream tyrosyl-tRNA synthetase gene. This study is the first description of a tyrosine decarboxylase gene in prokaryotes. PMID- 12089040 TI - Binding rather than metabolism may explain the interaction of two food-Grade Lactobacillus strains with zearalenone and its derivative (')alpha-earalenol. AB - The interaction between two Fusarium mycotoxins, zearalenone (ZEN) and its derivative (')alpha-zearalenol ((')alpha-ZOL), with two food-grade strains of Lactobacillus was investigated. The mycotoxins (2 microg ml(-1)) were incubated with either Lactobacillus rhamnosus strain GG or L. rhamnosus strain LC705. A considerable proportion (38 to 46%) of both toxins was recovered from the bacterial pellet, and no degradation products of ZEN and (')alpha-ZOL were detected in the high-performance liquid chromatograms of the supernatant of the culturing media and the methanol extract of the pellet. Both heat-treated and acid-treated bacteria were capable of removing the toxins, indicating that binding, not metabolism, is the mechanism by which the toxins are removed from the media. Binding of ZEN or (')alpha-ZOL by lyophilized L. rhamnosus GG and L. rhamnosus LC705 was a rapid reaction: approximately 55% of the toxins were bound instantly after mixing with the bacteria. Binding was dependent on the bacterial concentration, and coincubation of ZEN with (')alpha-ZOL significantly affected the percentage of the toxin bound, indicating that these toxins may share the same binding site on the bacterial surface. These results can be exploited in developing a new approach for detoxification of mycotoxins from foods and feeds. PMID- 12089041 TI - Silencing of the aspergillopepsin B (pepB) gene of Aspergillus awamori by antisense RNA expression or protease removal by gene disruption results in a large increase in thaumatin production. AB - Aspergillopepsin B was identified in culture broths of Aspergillus awamori by in situ detection of its proteolytic activity and by immunodetection with anti aspergillopepsin B antibodies. Severe thaumatin degradation was observed after in vitro treatment of thaumatin with purified aspergillopepsin B. The pepB gene encoding aspergillopepsin B of A. awamori was cloned and characterized. It is located in chromosome IV of A. awamori, as shown by pulsed-field gel electrophoresis, and encodes a protein of 282 amino acids with high similarity to the aspergillopepsin B of Aspergillus niger var. macrosporus. The pepB gene is expressed at high rates as a monocistronic 1.0-kb transcript in media with casein at acidic pH values. An antisense cassette constructed by inserting the pepB gene in the antisense orientation downstream from the gpdA promoter resulted in a good level of antisense mRNA, as shown by reverse transcription-PCR. Partial silencing of the pepB gene by the antisense mRNA resulted in a 31% increase in thaumatin yield. However, significant residual degradation of thaumatin still occurred. To completely remove aspergillopepsin B, the pepB gene was deleted by double crossover. Two of the selected transformants lacked the endogenous pepB gene and did not form aspergillopepsin B. Thaumatin yields increased by between 45% in transformant APB 7/25 and 125% in transformant 7/36 with respect to the parental strain. Reduction of proteolytic degradation by gene silencing with antisense mRNA or total removal of the aspergillopepsin B by directed gene deletion was a very useful method for improving thaumatin production in A. awamori. PMID- 12089042 TI - Source of enterococci in a farmhouse raw-milk cheese. AB - Enterococci are widely distributed in raw-milk cheeses and are generally thought to positively affect flavor development. Their natural habitats are the human and animal intestinal tracts, but they are also found in soil, on plants, and in the intestines of insects and birds. The source of enterococci in raw-milk cheese is unknown. In the present study, an epidemiological approach with pulsed-field gel electrophoresis (PFGE) was used to type 646 Enterococcus strains which were isolated from a Cheddar-type cheese, the milk it was made from, the feces of cows and humans associated with the cheese-making unit, and the environment, including the milking equipment, the water used on the farm, and the cows' teats. Nine different PFGE patterns, three of Enterococcus casseliflavus, five of Enterococcus faecalis, and one of Enterococcus durans, were found. The same three clones, one of E. faecalis and two of E. casseliflavus, dominated almost all of the milk, cheese, and human fecal samples. The two E. casseliflavus clones were also found in the bulk tank and the milking machine even after chlorination, suggesting that a niche where enterococci could grow was present and that contamination with enterococci begins with the milking equipment. It is likely but unproven that the enterococci present in the human feces are due to consumption of the cheese. Cow feces were not considered the source of enterococci in the cheese, as Enterococcus faecium and Streptococcus bovis, which largely dominated the cows' intestinal tracts, were not found in either the milk or the cheese. PMID- 12089043 TI - Cardiovascular effects of fermented milk containing angiotensin-converting enzyme inhibitors evaluated in permanently catheterized, spontaneously hypertensive rats. AB - In this study, two strains of Lactobacillus helveticus were used to produce fermented milk rich in angiotensin-converting enzyme (ACE) inhibitors. In vitro tests revealed that the two milks contained competitive inhibitors of ACE in amounts comparable to what has been obtained in previously reported studies. The two milks were administered by gavage to spontaneously hypertensive rats that had had a permanent aortic catheter inserted through the left arteria carotis, and mean arterial blood pressure and heart rate were monitored from 4 to 8 h after administration. Unfermented milk and milk fermented with a lactococcal strain that does not produce inhibitors were used as controls. Highly significant blood pressure effects were observed; i.e., milk fermented with the two strains of L. helveticus gave a more pronounced drop in blood pressure than the controls. Significant differences in heart rate effects were detected with one of the strains. PMID- 12089045 TI - Distribution of microbes producing antimicrobial epsilon-poly-L-lysine polymers in soil microflora determined by a novel method. AB - We developed a simple and sensitive screening method to investigate the distribution of microbes producing an antimicrobial poly(amino acid), epsilon poly-L-lysine (epsilon-PL), in microflora. An acidic dye, Poly R-478, incorporated in an agar plate detected epsilon-PL producers by electrostatic interaction with the secreted basic polymers. All epsilon-PL producers, isolated after careful and sufficient screening of soil microflora, belonged exclusively to two groups of bacteria of the family Streptomycetaceae and ergot fungi. They were characterized based on the density and diameter of the concentric zone formed by the secreted polymers. The density depended on each isolate. The increase in the diameter of the concentric zone per unit of time varied among isolates and was negatively correlated with the molecular weight. Although the distribution of epsilon-PL producers was extremely limited, their products were structurally varied. The molecular masses of the secreted polymers among the isolates ranged from 0.8 to 2.0 kDa. There were also isolates producing unknown polymers inconsistent with the correlation or producing a mixture of polymers with original and modified structures. A chemically modified polymer was an epsilon-PL derivative, as determined by mass spectrometry. Since the structural variations had no relation to the phylogenetic position of the isolates, it is possible that enzymes involved in the synthesis diversified after putative horizontal transfers of relevant genes. PMID- 12089044 TI - Lactobacillus casei is able to survive and initiate protein synthesis during its transit in the digestive tract of human flora-associated mice. AB - Live Lactobacillus casei is present in fermented dairy products and has beneficial properties for human health. In the human digestive tract, the resident flora generally prevents the establishment of ingested lactic acid bacteria, the presence of which is therefore transient. The aim of this work was to determine if L. casei DN-114 001 survives during transit and how this bacterium behaves in the digestive environment. We used the human flora associated (HFA) mouse model. L. casei DN-114 001 was genetically modified by the introduction of erm and lux genes, encoding erythromycin resistance and luciferase, respectively. For this modified strain (DN-240 041), light emission related to luciferase expression could easily be detected in the contents of the digestive tract. When inoculated into the digestive tract of HFA mice, L. casei (DN-240 041) survives but is eliminated with the same kinetics as an inert transit marker, indicating that it does not establish itself. In pure culture of L. casei, luciferase activities were high in the exponential and early stationary growth phases but decreased to become undetectable 1 day after inoculation. Viability was only slightly reduced even after more than 5 days. After transit in HFA mice, luciferase activity was detected even when 5-day-old L. casei cultures were given to the mice. In culture, the luciferase activity could be restored after 0.5 to 7 h of incubation in fresh medium or milk containing glucose, unless protein synthesis was inhibited by the addition of chloramphenicol or rifampin. These results suggest that in HFA mice L. casei DN-240 041, and thus probably L. casei DN-114 001, is able to initiate new protein synthesis during its transit with the diet. The beneficial properties of L. casei-fermented milk for human health might be related to this protein synthesis in the digestive tract. PMID- 12089046 TI - Biodegradation of 1,2,3-trichloropropane through directed evolution and heterologous expression of a haloalkane dehalogenase gene. AB - Using a combined strategy of random mutagenesis of haloalkane dehalogenase and genetic engineering of a chloropropanol-utilizing bacterium, we constructed an organism that is capable of growth on 1,2,3-trichloropropane (TCP). This highly toxic and recalcitrant compound is a waste product generated from the manufacture of the industrial chemical epichlorohydrin. Attempts to select and enrich bacterial cultures that can degrade TCP from environmental samples have repeatedly been unsuccessful, prohibiting the development of a biological process for groundwater treatment. The critical step in the aerobic degradation of TCP is the initial dehalogenation to 2,3-dichloro-1-propanol. We used random mutagenesis and screening on eosin-methylene blue agar plates to improve the activity on TCP of the haloalkane dehalogenase from Rhodococcus sp. m15-3 (DhaA). A second generation mutant containing two amino acid substitutions, Cys176Tyr and Tyr273Phe, was nearly eight times more efficient in dehalogenating TCP than wild type dehalogenase. Molecular modeling of the mutant dehalogenase indicated that the Cys176Tyr mutation has a global effect on the active-site structure, allowing a more productive binding of TCP within the active site, which was further fine tuned by Tyr273Phe. The evolved haloalkane dehalogenase was expressed under control of a constitutive promoter in the 2,3-dichloro-1-propanol-utilizing bacterium Agrobacterium radiobacter AD1, and the resulting strain was able to utilize TCP as the sole carbon and energy source. These results demonstrated that directed evolution of a key catabolic enzyme and its subsequent recruitment by a suitable host organism can be used for the construction of bacteria for the degradation of a toxic and environmentally recalcitrant chemical. PMID- 12089047 TI - Site-specific recombination-based genetic system for reporting transient or low level gene expression. AB - We report here the construction, characterization, and application of a plasmid based genetic system that reports the expression of a target promoter by effecting an irreversible, heritable change in a bacterial cell. This system confers strong repression of the reporter gene gfp in the absence of target promoter expression and utilizes the site-specific recombination machinery of bacteriophage P22 to trigger high-level reporter gene expression in the original cell and its progeny after target gene induction. We demonstrate the effectiveness of this genetic system by tailoring it to indicate the availability of arabinose to the biological control agent Enterobacter cloacae JL1157 in culture and in the barley rhizosphere. The presence of bioavailable arabinose triggered the production of P22 excisionase and integrase from the reporter plasmid pAraLHB in JL1157, and this led to excision of the cI repressor gene, which is flanked by att sites, and the subsequent irreversible expression of gfp in the original cell and in its progeny. In culture, nearly 100% of an E. cloacae JL1157(pAraLHB) population expressed gfp after exposure to 6.5 to 65 microM arabinose for 3 h. We used this biosensor to demonstrate that arabinose was released from the seeds of several legumes and grass species during germination and from roots of barley seedlings grown hydroponically or in soil. When introduced into microcosms containing barley, the biosensor permitted the localization of arabinose along the roots. Arabinose was present near the root seed junction and on the seminal roots but was not detected at the root tips. This recombination-based reporter system should be useful for monitoring bacterial exposure to transient or low levels of specific molecules directly in the environment. PMID- 12089048 TI - Noninvasive quantitative measurement of bacterial growth in porous media under unsaturated-flow conditions. AB - Glucose-dependent growth of the luxCDABE reporter bacterium Pseudomonas fluorescens HK44 was monitored noninvasively in quartz sand under unsaturated flow conditions within a 45- by 56- by 1-cm two-dimensional light transmission chamber. The spatial and temporal development of growth were mapped daily over 7 days by quantifying salicylate-induced bioluminescence. A nonlinear model relating the rate of increase in light emission after salicylate exposure to microbial density successfully predicted growth over 4 orders of magnitude (r(2) = 0.95). Total model-predicted growth agreed with growth calculated from the mass balance of the system by using previously established growth parameters of HK44 (predicted, 1.2 x 10(12) cells; calculated, 1.7 x 10(12) cells). Colonization expanded in all directions from the inoculation region, including upward migration against the liquid flow. Both the daily rate of expansion of the colonized zone and the population density of the first day's growth in each newly colonized region remained relatively constant throughout the experiment. Nonetheless, substantial growth continued to occur on subsequent days in the older regions of the colonized zone. The proportion of daily potential growth that remained within the chamber declined progressively between days 2 and 7 (from 97 to 13%). A densely populated, anoxic region developed in the interior of the colonized zone even though the sand was unsaturated and fresh growth medium continued to flow through the colonized zone. These data illustrate the potential of a light transmission chamber, bioluminescent bacteria, and sensitive digital camera technology to noninvasively study real-time hydrology-microbiology interactions associated with unsaturated flow in porous media. PMID- 12089049 TI - Changes in bacterioplankton community structure and activity with depth in a eutrophic lake as revealed by 5S rRNA analysis. AB - The community structure of bacterioplankton was studied at different depths (0 to 25 m) of a temperate eutrophic lake (Lake Plusssee in northern Germany) by using comparative 5S rRNA analysis. The relative amounts of taxonomic groups were estimated from 5S rRNA bands separated by high-resolution electrophoresis. Comparison of partial 5S rRNA sequences enabled detection of changes in single taxa over space and during seasons. Overall, the bacterioplankton community was dominated by 3 to 14 abundant (>4% of the total 5S rRNA) taxa. In general, the number of 5S rRNA bands (i.e., the number of bacterial taxa) decreased with depth. In the fall, when thermal stratification and chemical stratification were much more pronounced than they were in the spring, the correlation between the depth layers and the community structure was more pronounced. Therefore, in the fall each layer had its own community structure; i.e., there were different community structures in the epilimnion, the metalimnion, and the hypolimnion. Only three 5S rRNA bands were detected in the hypolimnion during the fall, and one band accounted for about 70% of the total 5S rRNA. The sequences of individual 5S rRNA bands from the spring and fall were different for all size classes analyzed except two bands, one of which was identified as Comamonas acidivorans. In the overall analysis of the depth profiles, the diversity in the epilimnion contrasted with the reduced diversity of the bacterioplankton communities in the hypolimnion, and large differences occurred in the composition of the epilimnion at different seasons except for generalists like C. acidivorans. PMID- 12089050 TI - Fitness of Salmonella enterica serovar Thompson in the cilantro phyllosphere. AB - The epiphytic fitness of Salmonella enterica was assessed on cilantro plants by using a strain of S. enterica serovar Thompson that was linked to an outbreak resulting from cilantro. Salmonella serovar Thompson had the ability to colonize the surface of cilantro leaves, where it was detected by confocal laser scanning microscopy (CLSM) at high densities on the veins and in natural lesions. The population sizes of two common colonizers of plant surfaces, Pantoea agglomerans and Pseudomonas chlororaphis, were 10-fold higher than that of the human pathogen on cilantro incubated at 22 degrees C. However, Salmonella serovar Thompson achieved significantly higher population levels and accounted for a higher proportion of the total culturable bacterial flora on cilantro leaves when the plants were incubated at warm temperatures, such as 30 degrees C, after inoculation, indicating that the higher growth rates exhibited by Salmonella serovar Thompson at warm temperatures may increase the competitiveness of this organism in the phyllosphere. The tolerance of Salmonella serovar Thompson to dry conditions on plants at 60% relative humidity was at least equal to that of P. agglomerans and P. chlororaphis. Moreover, after exposure to low humidity on cilantro, Salmonella serovar Thompson recovered under high humidity to achieve its maximum population size in the cilantro phyllosphere. Visualization by CLSM of green fluorescent protein-tagged Salmonella serovar Thompson and dsRed-tagged P. agglomerans inoculated onto cilantro revealed that the human pathogen and the bacterial epiphyte formed large heterogeneous aggregates on the leaf surface. Our studies support the hypothesis that preharvest contamination of crops by S. enterica plays a role in outbreaks linked to fresh fruits and vegetables. PMID- 12089051 TI - New chromogenic agar medium for the identification of Candida spp. AB - A new chromogenic agar medium (Candida diagnostic agar [CDA]) for differentiation of Candida spp. is described. This medium is based on Sabouraud dextrose agar (Oxoid CM41) and contains (per liter) 40.0 g of glucose, 10.0 g of mycological peptone, and 15.0 g of agar along with a novel chromogenic glucosaminidase substrate, ammonium 4-(2-[4-(2-acetamido-2-deoxy-beta-D-glucopyranosyloxy)-3 methoxyphenyl]-vinyl)-1-(propan-3-yl-oate)-quinolium bromide (0.32 g liter(-1)). The glucosaminidase substrate in CDA was hydrolyzed by Candida albicans and Candida dubliniensis, yielding white colonies with deep-red spots on a yellow transparent background after 24 to 48 h of incubation at 37 degrees C. Colonies of Candida tropicalis and Candida kefyr were uniformly pink, and colonies of other Candida spp., including Candida glabrata and Candida parapsilosis, were white. CDA was evaluated by using 115 test strains of Candida spp. and other clinically important yeasts and was compared with two commercially available chromogenic agars (Candida ID agar [bioMerieux] and CHROMagar Candida [CHROMagar Company Ltd.]). On all three agars, colonies of C. albicans were not distinguished from colonies of C. dubliniensis. However, for the group containing C. albicans plus C. dubliniensis, both the sensitivity and the specificity of detection when CDA was used were 100%, compared with values of 97.6 and 100%, respectively, with CHROMagar Candida and 100 and 96.8%, respectively, with Candida ID agar. In addition, for the group containing C. tropicalis plus C. kefyr, the sensitivity and specificity of detection when CDA was used were also 100%, compared with 72.7 and 98.1%, respectively, with CHROMagar Candida. Candida ID agar did not differentiate C. tropicalis and C. kefyr strains but did differentiate members of a broader group (C. tropicalis, C. kefyr, Candida lusitaniae plus Candida guilliermondii); the sensitivity and specificity of detection for members of this group were 94.7 and 93.8%, respectively. In addition to the increased sensitivity and/or specificity of Candida detection when CDA was used, differentiation of colony types on CDA (red spotted, pink, or no color) was unambiguous and did not require precise assessment of colony color. PMID- 12089052 TI - Use of 16S ribosomal DNA for delineation of marine bacterioplankton species. AB - All of the marine bacterioplankton-derived 16S ribosomal DNA sequences previously deposited in GenBank were reanalyzed to determine the number of bacterial species in the oceanic surface waters. These sequences have been entered into the database since 1990. The rate of new additions reached a peak in 1999 and subsequently leveled off, suggesting that much of the marine microbial species richness has been sampled. When the GenBank sequences were dereplicated by using 97% similarity as a cutoff, 1,117 unique ribotypes were found. Of the unique sequences, 609 came from uncultured environmental clones and 508 came from cultured bacteria. We conclude that the apparent bacterioplankton species richness is relatively low. PMID- 12089053 TI - Enrichment double-antibody sandwich indirect enzyme-linked immunosorbent assay that uses a specific monoclonal antibody for sensitive detection of Ralstonia solanacearum in asymptomatic potato tubers. AB - Sensitive and specific routine detection of Ralstonia solanacearum in symptomless potato tubers was achieved by efficient enrichment followed by a reliable double antibody sandwich indirect enzyme-linked immunosorbent assay based on the specific monoclonal antibody 8B-IVIA. This monoclonal antibody reacted with 168 typical R. solanacearum strains and did not recognize 174 other pathogenic or unidentified bacteria isolated from potato. The optimized protocol included an initial enrichment step consisting of shaking the samples in modified Wilbrink broth for 72 h at 29 degrees C. This step enabled specific detection by the enzyme-linked immunosorbent assay of 1 to 10 CFU of R. solanacearum per ml of initial potato extract. Analysis of 233 commercial potato lots by this method provided results that coincided with the results of conventional methods. PMID- 12089054 TI - Evidence of association of salmonellae with tomato plants grown hydroponically in inoculated nutrient solution. AB - The possibility of uptake of salmonellae by roots of hydroponically grown tomato plants was investigated. Within 1 day of exposure of plant roots to Hoagland nutrient solution containing 4.46 to 4.65 log(10) CFU of salmonellae/ml, the sizes of the pathogen populations were 3.01 CFU/g of hypocotyls and cotyledons and 3.40 log(10) CFU/g of stems for plants with intact root systems (control) and 2.55 log(10) CFU/g of hypocotyls and cotyledons for plants from which portions of the roots had been removed. A population of > or =3.38 log(10) CFU/g of hypocotyls-cotyledons, stems, and leaves of plants grown for 9 days was detected regardless of the root condition. Additional studies need to be done to unequivocally demonstrate that salmonellae can exist as endophytes in tomato plants grown under conditions that simulate commonly used agronomic practices. PMID- 12089055 TI - Phylogenetic relationships and coaggregation ability of freshwater biofilm bacteria. AB - Nineteen numerically dominant heterotrophic bacteria from a freshwater biofilm were identified by 16S ribosomal DNA gene sequencing, and their coaggregation partnerships were determined. Phylogenetic trees showed that both distantly related and closely related strains coaggregated at intergeneric, intrageneric, and intraspecies levels. One strain, Blastomonas natatoria 2.1, coaggregated with all 18 other strains and may function as a bridging organism in biofilm development. PMID- 12089056 TI - Cloning of a gene encoding raw-starch-digesting amylase from a Cytophaga sp. and its expression in Escherichia coli. AB - A raw-starch-digesting amylase (RSDA) gene from a Cytophaga sp. was cloned and sequenced. The predicted protein product contained 519 amino acids and had high amino acid identity to alpha-amylases from three Bacillus species. Only one of the Bacillus alpha-amylases has raw-starch-digesting capability, however. The RSDA, expressed in Escherichia coli, had properties similar to those of the enzyme purified from the Cytophaga sp. PMID- 12089057 TI - Differentiation of Paenibacillus larvae subsp. larvae, the cause of American foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNA. AB - A rapid procedure for the identification of Paenibacillus larvae subsp. larvae, the causal agent of American foulbrood (AFB) disease of honeybees (Apis mellifera L.), based on PCR and restriction fragment analysis of the 16S rRNA genes (rDNA) is described. Eighty-six bacterial strains belonging to 39 species of the genera Paenibacillus, Bacillus, Brevibacillus, and Virgibacillus were characterized. Amplified rDNA was digested with seven restriction endonucleases. The combined data from restriction analysis enabled us to distinguish 35 profiles. Cluster analysis revealed that P. larvae subsp. larvae and Paenibacillus larvae subsp. pulvifaciens formed a group with about 90% similarity; however, the P. larvae subsp. larvae restriction fragment length polymorphism pattern produced by endonuclease HaeIII was found to be unique and distinguishable among other closely related bacteria. This pattern was associated with DNA extracted directly from honeybee brood samples showing positive AFB clinical signs that yielded the restriction profile characteristic of P. larvae subsp. larvae, while no amplification product was obtained from healthy larvae. The method described here is particularly useful because of the short time required to carry it out and because it allows the differentiation of P. larvae subsp. larvae-infected larvae from all other species found in apiarian sources. PMID- 12089058 TI - Transcription factor decoy. PMID- 12089059 TI - Preventing apoptosis with thioredoxin: ASK me how. PMID- 12089060 TI - Importance of integrin signaling in myocyte growth and survival. PMID- 12089061 TI - Role of Akt signaling in vascular homeostasis and angiogenesis. AB - Akt is a serine/threonine protein kinase that is activated by a number of growth factors and cytokines in a phosphatidylinositol-3 kinase-dependent manner. Although antiapoptotic activity of Akt is well known, it also regulates other aspects of cellular functions, including migration, glucose metabolism, and protein synthesis. In this review, Akt signaling in endothelial cells and its critical roles in the regulation of vascular homeostasis and angiogenesis will be discussed. PMID- 12089062 TI - Nip21 gene expression reduces coxsackievirus B3 replication by promoting apoptotic cell death via a mitochondria-dependent pathway. AB - Our previous studies, using differential mRNA display, suggested that the mouse Nip21 gene may be involved in myocarditis development in the coxsackievirus B3 (CVB3)-infected mouse heart. Sequence comparison indicated that the mouse Nip21 gene shares high sequence homology to human Nip2. This human protein is known to interact with both the apoptosis inhibitor Bcl-2 and a homologous protein, the adenovirus E1B 19-kDa protein. Such interactions implicate Nip21 gene in cell death pathways. To study the function of this gene, we have cloned Nip21 from mouse hearts and established a Tet-On doxycycline-inducible HeLa cell line and a cardiomyocyte H9c2 cell line expressing Nip21 to characterize gene function in relation to apoptosis. We demonstrated that Nip21 expression could induce apoptosis via caspase-depended mitochondria activation. To further determine the function of Nip21 in CVB3-induced apoptosis, the Tet-On/Nip21 HeLa cell line was induced by doxycycline followed by CVB3 infection. We found that activation of caspase-3 and cleavage of poly-(ADP-ribose) polymerase occurred 2 hours earlier than in vector-transfected control cells, suggesting that Nip21 expression enhances CVB3-induced apoptosis. We also demonstrated a significant decrease in HeLa cell and H9c2 cell viability. Particularly, as illustrated by viral plaque assay, CVB3 replication was dramatically reduced in Tet-On HeLa cells, due at least in part to the earlier killing of the host cells by Nip21 overexpression. PMID- 12089063 TI - Thioredoxin promotes ASK1 ubiquitination and degradation to inhibit ASK1-mediated apoptosis in a redox activity-independent manner. AB - It has been shown that thioredoxin (Trx) in a reduced form binds to and inhibits apoptosis signal-regulating kinase 1 (ASK1). Apoptotic stimuli such as tumor necrosis factor (TNF) and reactive oxygen species (ROS) activate ASK1 in part by oxidizing Trx (forming intramolecular disulfide between C32 and C35) to release Trx from ASK1. In the present study, we examined if Trx affects ASK1 protein stability and whether the redox activity of Trx is critical in regulating ASK1 activity. First, we showed that overexpression of the wild-type Trx (Trx-WT) in endothelial cells induced ASK1 ubiquitination and degradation. Trx-induced ASK1 ubiquitination/degradation could be blocked by ASK1 activators TNF and TRAF2. We then tested the single-mutation of Trx at the catalytic site C32 or C35 (Trx-C32S or Trx-C35S) and the double-mutation (Trx-CS). The results showed that the single mutants (but not Trx-CS) retained the binding activity for ASK1 and the ability to induce ASK1 ubiquitination/degradation. Unlike Trx-WT, Trx-C32S and Trx-C35S mutants constitutively bind to ASK1 even in the presence of hydrogen peroxide in vitro and TNF in vivo. Finally, we showed that the single mutants (not Trx-WT) significantly (n=4 and P<0.05) inhibited ASK1-induced JNK activation, caspase 3 activity, and apoptosis in TNF/ROS-resistant manner. Our data suggest that association of Trx with ASK1 through a single Cysteine (C32 or C35) is necessary and sufficient for Trx activity in inducing ASK1 ubiquitination/degradation leading to inhibition of ASK1-induced apoptosis. PMID- 12089064 TI - Dominant-negative suppression of HCN1- and HCN2-encoded pacemaker currents by an engineered HCN1 construct: insights into structure-function relationships and multimerization. AB - I(f), a diastolic depolarizing current activated by hyperpolarization, is a key player in cardiac pacing. Despite the fact that I(f) has been known for over 20 years, the encoding genes, namely HCN1 to 4, have only recently been identified. Functional data imply that different HCN isoforms may coassemble to form heteromeric channel complexes, but little direct evidence is available. Subunit stoichiometry is also unknown. Although the pore region of HCN channels contains the glycine-tyrosine-glycine (GYG) signature motif found in K+-selective channels, they permeate both Na+ and K+. In the present study, we probed the functional importance of the GYG selectivity motif in pacemaker channels by replacing this triplet in HCN1 with alanines (GYG(349-351)AAA or HCN1-AAA). HCN1 AAA did not yield functional currents; coexpression of HCN1-AAA with wild-type (WT) HCN1 suppressed normal channel activity in a dominant-negative manner (55.2+/-3.2%, 68.3+/-4.3%, 78.7+/-1.6%, 91.7+/-0.8%, and 97.9+/-0.2% current reduction at -140 mV for WT:AAA cRNA ratios of 4:1, 3:1, 2:1, 1:1, and 1:2, respectively) without affecting gating (steady-state activation, activation and deactivation kinetics) or permeation (reversal potential) properties. HCN1-AAA coexpression, however, did not alter the expressed current amplitudes of Kv1.4 and Kv2.1 channels, indicating that its suppressive effect was channel-specific. Statistical analysis reveals that a single HCN channel is composed of 4 monomeric subunits. Interestingly, HCN1-AAA also inhibited HCN2 in a dominant-negative manner with the same efficacy. We conclude that the GYG motif is a critical determinant of ion permeation for HCN channels, and that HCN1 and HCN2 readily coassemble to form heterotetrameric complexes. PMID- 12089065 TI - Hypoxia-reoxygenation: a potent inducer of apoptotic changes in the human placenta and possible etiological factor in preeclampsia. AB - Preeclampsia is a severe disorder of human pregnancy characterized by generalized activation of maternal endothelial cells. Oxidative stress of the placenta is considered a key intermediary step, precipitating deportation of apoptotic fragments into the maternal circulation, but the cause remains unknown. We hypothesize that intermittent placental perfusion, secondary to deficient trophoblast invasion of the endometrial arteries, leads to an ischemia reperfusion-type insult. We therefore tested whether hypoxia-reoxygenation (H/R) in vitro stimulates apoptosis in human placental tissues compared with controls kept hypoxic or normoxic throughout. After H/R, release of cytochrome c from mitochondria was significantly increased and was associated with intense immunolabeling for active caspase 3 in the syncytiotrophoblast and fetal endothelial cells. There was also increased labeling of syncytiotrophoblastic nuclei for cleaved poly (ADP-ribose) polymerase (PARP), and higher cytosolic concentrations of cleaved PARP fragment were detected by Western blot. Syncytiotrophoblastic nuclei displayed increased chromatin condensation, and a significantly greater percentage was TUNEL positive. These changes were accompanied by increased lactate dehydrogenase release into the medium. Preadministration of the free radical scavenger, desferrioxamine, reduced cytochrome c release and the TUNEL-positive index, suggesting generation of hydroxyl radicals mediates these processes. By contrast, hypoxia alone caused a smaller increase in the TUNEL-positive index, and the majority of syncytiotrophoblastic nuclei displayed karyolysis, whereas normoxic controls remained euchromatic. We conclude that H/R stimulates apoptotic changes within the syncytiotrophoblast, whereas hypoxia principally induces necrosis. The quality of placental perfusion may therefore be a more important factor in the pathophysiology of preeclampsia than the absolute quantity. PMID- 12089066 TI - GFP-FRNK disrupts focal adhesions and induces anoikis in neonatal rat ventricular myocytes. AB - Focal adhesion kinase (FAK) is a nonreceptor protein tyrosine kinase involved in adhesion-dependent signal transduction. FAK is highly expressed in cultured neonatal rat ventricular myocytes (NRVMs) and undergoes tyrosine autophosphorylation in response to cell adhesion, stretch, and growth factor stimulation. We previously showed that inhibition of FAK phosphorylation by adenovirally mediated overexpression of FRNK (the autonomously expressed C terminal domain of FAK) prevented endothelin-1 (ET)-induced NRVM hypertrophy. One question raised by these studies was whether FRNK localized to focal adhesions and displaced FAK from sites required for downstream signaling. Therefore, we constructed a replication-defective adenovirus encoding a GFP-FRNK fusion protein (Adv-GFP-FRNK) and examined its effects on NRVM cytoarchitecture and signaling. Uninfected NRVMs contained small amounts of endogenous FRNK. NRVMs infected with Adv-GFP-FRNK expressed much larger amounts of a 66-/68-kDa protein that localized to costameres and focal adhesions. GFP-FRNK overexpression suppressed basal and ET-induced FAK phosphorylation and also inhibited ET-induced phosphorylation of PYK2, the other member of the FAK family of nonreceptor protein tyrosine kinases. In contrast, GFP-FRNK overexpression did not prevent ET-induced ERK, JNK, or p70S6K phosphorylation. Furthermore, GFP-FRNK resulted in the loss of detectable FAK and paxillin in focal adhesions, which was accompanied by reduced levels of total paxillin and, ultimately, cell detachment and apoptosis. We conclude that FRNK functions as a dominant-negative inhibitor of adhesion-dependent signaling by displacing FAK from focal adhesions and interfering with the anchorage of NRVMs that is necessary for cell survival, a process known as anoikis. PMID- 12089067 TI - Glycated collagen I induces premature senescence-like phenotypic changes in endothelial cells. AB - Diabetic vasculopathy is central to the development of diverse cardiovascular, renal, retinal, and neurological complications of diabetes. We previously demonstrated that growth of endothelial cells on glycated extracellular matrix proteins (collagen and matrigel) results in a significant decrease in cell proliferation. In the present study, we show that early-passage human umbilical vein endothelial cells (HUVECs) grown on glycated collagen (GC) express hallmarks of premature cell senescence, ie, increase in the proportion of cells expressing senescence-associated beta-galactosidase activity, apoptotic rate, and p53 and p14(AFR) expression, but in contrast to replicative senescence, display neither attrition of telomeres nor decrease in telomerase activity. An increased frequency of prematurely senescent cells was similarly observed in vivo in aortae of young Zucker diabetic rats, compared with lean controls. NO production by HUVECs grown on GC was decreased, despite a 3-fold increase in eNOS expression and was associated with the increased nitrotyrosine-modified proteins. Development of premature senescence of HUVECs on GC could be prevented and reversed by treatments with the peroxynitrite scavenger, ebselen, eNOS intermediate N(omega)-hydroxy-L-arginine (NOHA), or superoxide dismutase mimetic Mn-TBAP. Concomitant with the reversal of senescence, ebselen, and NOHA each restored NO production to levels observed with HUVECs grown on unmodified collagen. Our findings indicate that diabetes mellitus in vivo and GC exposure in vitro elicit premature senescence of the vascular endothelium, a process with distinct pathogenetic mechanisms. Premature senescence of the vascular endothelium is hypothesized to be an important contributor to diabetic vasculopathy and a consequence of reduced NO availability, peroxynitrite, and/or superoxide excess. PMID- 12089068 TI - Actin capping protein: an essential element in protein kinase signaling to the myofilaments. AB - Actin capping protein (CapZ) binds the barbed ends of actin at sarcomeric Z lines. In addition to anchoring actin, Z-discs bind protein kinase C (PKC). Although CapZ is crucial for myofibrillogenesis, its role in muscle function and intracellular signaling is unknown. We hypothesized that CapZ downregulation would impair myocardial function and disrupt PKC-myofilament signaling by impairing PKC-Z-disc interaction. To test these hypotheses, we examined transgenic (TG) mice in which cardiac CapZ protein is reduced. Fiber bundles were dissected from papillary muscles and detergent extracted. Some fiber bundles were treated with PKC activators phenylephrine (PHE) or endothelin (ET) before detergent extraction. We simultaneously measured Ca2+-dependent tension and actomyosin MgATPase activity. CapZ downregulation increased myofilament Ca2+ sensitivity without affecting maximum tension or actomyosin MgATPase activity. Maximum tension and actomyosin MgATPase activity were decreased after PHE or ET treatment of wild-type (WT) muscle. Fiber bundles from TG hearts did not respond to PHE or ET. Immunoblot analysis revealed an increase in myofilament-associated PKC-epsilon after PHE or ET exposure of WT preparations. In contrast, myofilament associated PKC-epsilon was decreased after PHE or ET treatment in TG myocardium. Protein levels of myofilament-associated PKC-beta were decreased in TG ventricle. C-protein and troponin I phosphorylation was increased after PHE or ET treatment in WT and TG hearts. Basal phosphorylation levels of C-protein and troponin I were higher in TG myocardium. These results indicate that downregulation of CapZ, or other changes associated with CapZ downregulation, increases cardiac myofilament Ca2+ sensitivity, inhibits PKC-mediated control of myofilament activation, and decreases myofilament-associated PKC-beta. PMID- 12089069 TI - Diversity in mitochondrial function explains differences in vascular oxygen sensing. AB - Renal arteries (RAs) dilate in response to hypoxia, whereas the pulmonary arteries (PAs) constrict. In the PA, O2 tension is detected by an unidentified redox sensor, which controls K+ channel function and thus smooth muscle cell (SMC) membrane potential and cytosolic calcium. Mitochondria are important regulators of cellular redox status and are candidate vascular O2 sensors. Mitochondria-derived activated oxygen species (AOS), like H2O2, can diffuse to the cytoplasm and cause vasodilatation by activating sarcolemmal K+ channels. We hypothesize that mitochondrial diversity between vascular beds explains the opposing responses to hypoxia in PAs versus RAs. The effects of hypoxia and proximal electron transport chain (pETC) inhibitors (rotenone and antimycin A) were compared in rat isolated arteries, vascular SMCs, and perfused organs. Hypoxia and pETC inhibitors decrease production of AOS and outward K+ current and constrict PAs while increasing AOS production and outward K+ current and dilating RAs. At baseline, lung mitochondria have lower respiratory rates and higher rates of AOS and H2O2 production. Similarly, production of AOS and H2O2 is greater in PA versus RA rings. SMC mitochondrial membrane potential is more depolarized in PAs versus RAs. These differences relate in part to the lower expression of proximal ETC components and greater expression of mitochondrial manganese superoxide dismutase in PAs versus RAs. Differential regulation of a tonically produced, mitochondria-derived, vasodilating factor, possibly H2O2, can explain the opposing effects of hypoxia on the PAs versus RAs. We conclude that the PA and RA have different mitochondria. PMID- 12089070 TI - Renal myogenic response: kinetic attributes and physiological role. AB - The kinetic attributes of the afferent arteriole myogenic response were investigated using the in vitro perfused hydronephrotic rat kidney. Equations describing the time course for pressure-dependent vasoconstriction and vasodilation, and steady-state changes in diameter were combined to develop a mathematical model of autoregulation. Transfer functions were constructed by passing sinusoidal pressure waves through the model. These findings were compared with results derived using data from instrumented conscious rats. In each case, a reduction in gain and increase in phase were observed at frequencies of 0.2 to 0.3 Hz. We then examined the impact of oscillating pressure signals. The model predicted that pressure signals oscillating at frequencies above the myogenic operating range would elicit a sustained vasoconstriction the magnitude of which was dependent on peak pressure. These predictions were directly confirmed in the hydronephrotic kidney. Pressure oscillations presented at frequencies of 1 to 6 Hz elicited sustained afferent vasoconstrictions and the magnitude of the response depended exclusively on the peak pressure. Elevated systolic pressure elicited vasoconstriction even if mean pressure was reduced. These findings challenge the view that the renal myogenic response exists to maintain glomerular capillary pressure constant, but rather imply a primary role in protecting against elevated systolic pressures. Thus, the kinetic features of the afferent arteriole allow this vessel to adjust tone in response to changes in systolic pressures presented at the pulse rate. We suggest that the primary function of this mechanism is to protect the glomerulus from the blood pressure power that is normally present at the pulse frequency. PMID- 12089071 TI - Inhibitory effects of novel AP-1 decoy oligodeoxynucleotides on vascular smooth muscle cell proliferation in vitro and neointimal formation in vivo. AB - Excessive proliferation of vascular smooth muscle cells (VSMCs) and neointimal formation are critical steps in the pathogenesis of atherosclerosis and restenosis after percutaneous transluminal angioplasty. In this study, we investigated the hypothesis that the activator protein-1 (AP-1) plays an important role in neointimal formation after vascular injury. A circular dumbbell AP-1 decoy oligodeoxynucleotide (CDODN) was developed as a novel therapeutic strategy for restenosis after angioplasty. This CDODN was more stable than the conventional phosphorothioate linear decoy ODN (PSODN) and maintained structural integrity on exposure to exonuclease III or serum. Transfection with AP-1 decoy ODNs strongly inhibited VSMC proliferation and migration, as well as glucose- and serum-induced expression of PCNA and cyclin A genes. Administration of AP-1 decoy ODNs in vivo using the hemagglutinating virus of Japan (HVJ)-liposome method virtually abolished neointimal formation after balloon injury to the rat carotid artery. Compared with PSODN, CDODN was more effective in inhibiting the proliferation of VSMCs in vitro and neointimal formation in vivo. Our results collectively indicate that AP-1 activation is crucial for the mediation of VSMC proliferation in response to vascular injury. Moreover, the use of stable CDODN specific for AP-1 activity in combination with the highly effective HVJ-liposome method provides a novel potential therapeutic strategy for the prevention of restenosis after angioplasty in humans. PMID- 12089072 TI - Apolipoprotein E mediates the retention of high-density lipoproteins by mouse carotid arteries and cultured arterial smooth muscle cell extracellular matrices. AB - Lipoprotein retention in the vascular extracellular matrix (ECM) plays a critical role in atherogenesis. Previous studies demonstrated the presence of apo A-I and E in atherosclerotic lesions, suggesting that HDL may be trapped by the artery wall. We sought to determine mechanisms by which HDL could be bound and retained by the arterial wall, and whether apo E was a principal determinant of this binding. We evaluated in situ accumulation of fluorescently labeled DiI-human HDL+/-apo E in perfused carotid arteries from apo E-null mice. Apo E was important in mediating HDL binding to the vascular wall, with a 48+/-16% increase in accumulation of DiI-labeled apo E-containing HDL (HDL3+E) compared with DiI apo E-free HDL (HDL3-E) (P=0.003). To investigate possible mechanisms responsible for retention, we assessed binding of unlabeled HDL3-E and HDL3+E to ECM generated by cultured arterial smooth muscle cells. Similar to the in situ carotid artery data, HDL3+E bound better to the ECM than did HDL3-E (3-fold lower K(a) and 3.5-fold higher B(max) for HDL3+E versus HDL3-E). These differences were eliminated after either neutralization of arginine residues on apo E or digestion of matrix with chondroitin ABC lyase, suggesting that chondroitin and/or dermatan sulfate proteoglycans were responsible for apo E-mediated increased binding. These findings demonstrate that HDL can bind to both intact murine carotid arteries and smooth muscle cell-derived ECM, and that apo E is a principal determinant in mediating the ability of HDL to be trapped and retained via its interaction with ECM proteoglycans. PMID- 12089073 TI - New treatments for atopic dermatitis. PMID- 12089074 TI - Long term care for older people. PMID- 12089075 TI - Psychological medicine. PMID- 12089076 TI - Whooping cough--a continuing problem. PMID- 12089078 TI - Doctor "interfered" in treatment of girl after parents rejected his help. PMID- 12089080 TI - US draws up plans for smallpox outbreak after terrorist attack. PMID- 12089081 TI - Give patients a second chance before striking them off, says ombudsman. PMID- 12089082 TI - AMA considers whether to pay for donation of organs. PMID- 12089083 TI - BMA has to pay 815000 pounds sterling in damages for indirect racial discrimination. PMID- 12089087 TI - Doped East German athletes to receive compensation. PMID- 12089089 TI - Anti-leukotrienes as add-on therapy to inhaled glucocorticoids in patients with asthma: systematic review of current evidence. AB - OBJECTIVES: To examine the evidence for the efficacy and glucocorticoid sparing effect of oral anti-leukotrienes taken daily as add-on therapy to inhaled glucocorticoids in patients with asthma. DESIGN: Systematic review of randomised controlled trials of children and adults with asthma comparing the addition of anti-leukotrienes or placebo to inhaled glucocorticoids. MAIN OUTCOME MEASURES: The rate of exacerbations of asthma requiring rescue systemic glucocorticoids when the intervention was compared to the same or double dose of inhaled glucocorticoids, and the glucocorticoid sparing effect when the intervention was aimed at tapering the glucocorticoid. RESULTS: Of 376 citations, 13 were included: 12 in adult patients and one in children. The addition of licensed doses of anti-leukotrienes to inhaled glucocorticoids resulted in a non significant reduction in the risk of exacerbations requiring systemic steroids (two trials; relative risk 0.61, 95% confidence interval 0.36 to 1.05). No trials comparing the use of anti-leukotrienes with double the dose of inhaled glucocorticoids could be pooled. The use of anti-leukotrienes resulted in no overall group difference in the lowest achieved dose of inhaled glucocorticoids (three trials; weighted mean difference -44.43 microg/day, -147.87 to 59.02: random effect model) but was associated with a reduction in withdrawals owing to poor asthma control (four trials; relative risk 0.56, 0.35 to 0.89). CONCLUSIONS: The addition of anti-leukotrienes to inhaled glucocorticoids may modestly improve asthma control compared with inhaled glucocorticoids alone but this strategy cannot be recommended as a substitute for increasing the dose of inhaled glucocorticoids. The addition of anti-leukotrienes is possibly associated with superior asthma control after tapering of glucocorticoids, but the glucocorticoids sparing effect cannot be quantified at present. PMID- 12089090 TI - Amblyopia treatment outcomes after screening before or at age 3 years: follow up from randomised trial. AB - OBJECTIVE: To assess the effectiveness of early treatment for amblyopia in children. DESIGN: Follow up of outcomes of treatment for amblyopia in a randomised controlled trial comparing intensive orthoptic screening at 8, 12, 18, 25, 31, and 37 months (intensive group) with orthoptic screening at 37 months only (control group). SETTING: Avon, southwest England. PARTICIPANTS: 3490 children who were part of a birth cohort study. MAIN OUTCOME MEASURES: Prevalence of amblyopia and visual acuity of the worse seeing eye at 7.5 years of age. RESULTS: Amblyopia at 7.5 years was less prevalent in the intensive group than in the control group (0.6% v 1.8%; P=0.02). Mean visual acuities in the worse seeing eye were better for children who had been treated for amblyopia in the intensive group than for similar children in the control group (0.15 v 0.26 LogMAR units; P<0.001). A higher proportion of the children who were treated for amblyopia had been seen in a hospital eye clinic before 3 years of age in the intensive group than in the control group (48% v 13%; P=0.0002). CONCLUSIONS: The intensive screening protocol was associated with better acuity in the amblyopic eye and a lower prevalence of amblyopia at 7.5 years of age, in comparison with screening at 37 months only. These data support the hypothesis that early treatment for amblyopia leads to a better outcome than later treatment and may act as a stimulus for research into feasible screening programmes. PMID- 12089091 TI - Spiritual beliefs may affect outcome of bereavement: prospective study. AB - OBJECTIVE: To explore the relation between spiritual beliefs and resolution of bereavement. DESIGN: Prospective cohort study of people about to be bereaved with follow up continuing for 14 months after the death. SETTING: A Marie Curie centre for specialist palliative care in London. PARTICIPANTS: 135 relatives and close friends of patients admitted to the centre with terminal illness. MAIN OUTCOME MEASURE: Core bereavement items, a standardised measure of grief, measured 1, 9, and 14 months after the patients' death. RESULTS: People reporting no spiritual belief had not resolved their grief by 14 months after the death. Participants with strong spiritual beliefs resolved their grief progressively over the same period. People with low levels of belief showed little change in the first nine months but thereafter resolved their grief. These differences approached significance in a repeated measures analysis of variance (F=2.42, P=0.058). Strength of spiritual belief remained an important predictor after the explanatory power of relevant confounding variables was controlled for. At 14 months the difference between the group with no beliefs and the combined low and high belief groups was 7.30 (95% confidence interval 0.86 to 13.73) points on the core bereavement items scale. Adjusting for confounders in the final model reduced this difference to 4.64 (1.04 to 10.32) points. CONCLUSION: People who profess stronger spiritual beliefs seem to resolve their grief more rapidly and completely after the death of a close person than do people with no spiritual beliefs. PMID- 12089092 TI - Medical students' compliance with simple administrative tasks and success in final examinations: retrospective cohort study. PMID- 12089093 TI - Prolonged erections produced by dihydrocodeine and sildenafil. PMID- 12089094 TI - An observational study comparing quality of care in walk-in centres with general practice and NHS Direct using standardised patients. AB - OBJECTIVES: To compare the quality of clinical care in walk-in centres with that provided in general practice and by NHS Direct. DESIGN: Observational study involving assessment of clinicians by standardised patients. SETTING: 20 walk-in centres, 20 general practices, and 11 NHS Direct sites. PARTICIPANTS: 297 consultations with standardised patients, 99 in each setting, carried out by professional role players trained to play five clinical scenarios (postcoital contraception, chest pain, sinusitis, headache, and asthma). MAIN OUTCOME MEASURES: Primary outcomes were mean scores on consensus derived checklists of essential items for the management of the clinical scenarios. Data were also collected on access to and referral by walk-in centres, general practices, and NHS Direct. RESULTS: Walk-in centres achieved a significantly greater mean score for all scenarios combined than general practices (difference between groups 8.2, 95% confidence interval 1.7 to 14.6) and NHS Direct (10.8, 5.5 to 16.1). There was considerable between scenario variation, with walk-in centres performing particularly well on postcoital contraception and asthma scenarios. In contrast to general practices, walk-in centres and NHS Direct referred a higher proportion of patients (26% and 82%, respectively). CONCLUSION: Walk-in centres perform adequately and safely compared with general practices and NHS Direct for the range of conditions under study, but the impact of referrals on workload of other healthcare providers requires further research. PMID- 12089095 TI - Research in medical education: three decades of progress. PMID- 12089096 TI - Dengue: an escalating problem. PMID- 12089097 TI - ABC of psychological medicine. The consultation. PMID- 12089098 TI - Risk factor thresholds: their existence under scrutiny. PMID- 12089099 TI - Effective diabetes care: a need for realistic targets. PMID- 12089100 TI - Alteplase for stroke. Uncertainty remains about efficacy. PMID- 12089101 TI - Authors' reply to getting more for their dollar: Kaiser v the NHS. PMID- 12089102 TI - Health care in Britain remains variable. PMID- 12089104 TI - Ethics of limb allografts. Authors assume that they understand more than their patients do. PMID- 12089103 TI - PFI is here to stay. Select committee's report used parliamentary privilege unacceptably. PMID- 12089105 TI - Co-artemether has been used in ambulatory treatment of falciparum malaria. PMID- 12089106 TI - Action is still needed to stem heterosexual transmission of HIV in Africa. PMID- 12089107 TI - Clarifications: open access follow up for inflammatory bowel disease. PMID- 12089108 TI - Managed clinical networks. PMID- 12089110 TI - Shortage of vaccines: USA, Europe, who next? PMID- 12089109 TI - Newborn and childhood screening programmes: criteria, evidence, and current policy. PMID- 12089111 TI - Markers of microvascular complications in insulin dependent diabetes. PMID- 12089112 TI - Borrelia lymphocytoma ("lymphadenosis benigna cutis"). PMID- 12089113 TI - The meningococcus tamed? AB - Serogroup B Neisseria meningitidis is a frequent cause of invasive meningococcal disease, yet there are no effective vaccines suitable for routine immunisation. Limited efficacy has been shown with meningococcal outer membrane vacccines in children 4 years and older. Here we review the status of current research and consider new approaches to development of meningococcal serogroup B vaccines. PMID- 12089115 TI - Antivaccination activists on the world wide web. AB - AIMS: To determine the likelihood of finding an antivaccination site on the world wide web and to characterise their explicit claims and rhetorical appeals. METHODS: Using "vaccination" and "immunisation", examining the first 10 sites displayed on seven leading search engines. Detailed examination of content of 100 antivaccination sites found on Google. RESULTS: 43% of websites were antivaccination (all of the first 10 on Google). Main rhetorical appeals involve themes of the scientific veracity of antivaccination argument; rapport with parents seeking to protect their children from harm; and alleged collusion between doctors, the pharmaceutical industry, and government to deny vaccine harm. CONCLUSIONS: There is a high probability that parents will encounter elaborate antivaccination material on the world wide web. Factual refutational strategies alone are unlikely to counter the highly rhetorical appeals that shape these sites. PMID- 12089116 TI - Cycling with myth Australia. PMID- 12089114 TI - Should the new pneumococcal vaccine be used in high-risk children? AB - A new conjugate 7-valent vaccine to prevent pneumococcal infection (Prevenar, Wyeth) has recently received a European licence for use in young healthy children. The vaccine is not currently included in the universal immunisation schedule in the UK or elsewhere in Europe, although it is being used widely in the USA. Its availability for purchase raises the question whether paediatricians should consider using it in high risk children, including those for whom the polysaccharide 23-valent vaccine was previously recommended, until (or unless) it is introduced into general use-indeed the Chief Medical Officer for England and Wales has recently made a recommendation regarding such children aged less than 2 years. We review the evidence concerning use of the vaccine in such children and make suggestions as to how the vaccine may be used while further information is collected. PMID- 12089117 TI - Hygiene levels in a contemporary population cohort are associated with wheezing and atopic eczema in preschool infants. AB - BACKGROUND: The hygiene hypothesis states that insufficient exposure to certain infectious agents during childhood increases the risk of developing asthma and atopic diseases. Improvements in hygiene levels may be partly responsible for this decline in exposure. AIMS: To assess whether hygiene levels in infancy are associated with wheeze and/or atopic eczema, independent of a number of possible confounding factors. METHODS: Data were gathered from the Avon Longitudinal Study of Parents and Children (ALSPAC). Parental self completion questionnaires provided symptom data on infant wheeze and atopic eczema at 0-6 months and 30-42 months, respectively. A simple hygiene score was derived using questionnaire responses at 15 months, which ranged from least hygienic to most hygienic. Multivariable logistic regression models analysed the effect of hygiene scores on health outcomes, while adjusting for a number of important confounding variables. RESULTS: Increasing hygiene scores were independently associated with wheezing (OR = 1.04; 95% CI: 1.00 to 1.08) and atopic eczema (OR = 1.04; 95% CI: 1.01 to 1.07) between 30 and 42 months, but not in the first six months. The odds ratio was higher for atopic eczema if the rash was reported to have become sore and oozy (OR = 1.09; 95% CI: 1.02 to 1.16). CONCLUSIONS: High levels of hygiene at 15 months of age were independently associated with wheeze and atopic eczema reported between 30 and 42 months, and there was an increased risk for children with more severe eczema during this period. The importance of hygiene in public health should not be dismissed; however, the creation of a sterile environment through excessive cleanliness may potentially be harmful to the immune system. PMID- 12089119 TI - Bereavement support following sudden and unexpected death: guidelines for care. AB - Dealing with families who have suffered a sudden and unexpected death is a skill that may be needed by any paediatrician. Offering a bereavement follow up meeting to such families is part of accepted practice and is perceived to be of value in helping the family to come to terms with the loss. Unfortunately, there is very little guidance on the objectives for such a meeting, or the training required to help staff conduct such meetings. The nature of the work on a paediatric intensive care unit (PICU) means that staff have a greater experience of handling families in such a situation. We have reviewed our experience over the past five years following up the families of 51 children who have died suddenly and unexpectedly in our regional PICU. In doing this we have identified five key elements that we suggest are essential to a successful follow up meeting, and have supported this with case studies as illustration. PMID- 12089118 TI - Factors associated with different hygiene practices in the homes of 15 month old infants. AB - BACKGROUND: Improved hygiene in Westernised regions of the world may be partly responsible for the increased prevalence of diseases of the immune system, such as asthma and atopy. There is a paucity of data on cleanliness norms in young children in the UK and there has been no attempt to identify factors that influence the adoption of particular hygiene practices in the home. AIMS: To examine levels of hygiene in a contemporary cohort of children and identify social and lifestyle factors influencing hygiene practices in the home. METHODS: The sample under study are participants in the Avon Longitudinal Study of Parents and Children (ALSPAC). Parental self completion questionnaires provided data on hygiene levels in children at 15 months of age, and a hygiene score was derived from these responses. Multivariable logistic regression models investigated associations between high hygiene scores (top quintile) and a number of perinatal, maternal, social, and environmental factors. RESULTS: Maternal smoking during pregnancy, low maternal educational achievement, and living in local authority housing were factors independently associated with high hygiene scores, as was increased use of chemical household products. High hygiene scores were inversely related to living in damp housing and attendance at day care. There were no gender or ethnic differences in hygiene score. CONCLUSION: Important data on cleanliness norms for infants have been presented. The adoption of hygiene practices is influenced to some degree by social, lifestyle, and environmental factors-with higher hygiene scores occurring in more socially disadvantaged groups. Increased use of chemical household products in the more socially disadvantaged groups within ALSPAC has emerged as an important confounder in any study of hygiene and ill health. PMID- 12089121 TI - Should paediatric units have bereavement support posts? AB - Eight bereavement support posts in paediatrics and maternity were established using the experience of the Child Bereavement Trust (CBT). Evaluation showed that staff welcomed the informal support and teaching for themselves, as well as support for families; 67% of staff who returned questionnaires felt care for bereaved families had improved as a result of the posts. Many felt that there had been a positive effect on staff retention and stress levels. PMID- 12089122 TI - Excessive infant crying: definitions determine risk groups. AB - We assessed risk groups for excessive infant crying using 10 published definitions, in 3179 children aged 1-6 months (response: 96.5%). Risk groups regarding parental employment, living area, lifestyle, and obstetric history varied by definition. This may explain the existence of conflicting evidence on the occurrence and cause of excessive crying. PMID- 12089123 TI - Cushing's syndrome, growth impairment, and occult adrenal suppression associated with intranasal steroids. AB - We have previously described iatrogenic Cushing's syndrome secondary to intranasal steroids. This report further highlights the potential deleterious effects of intranasal steroids. Nine cases (including the original two cases) are reviewed to show the varied clinical manifestations of adrenal suppression caused by intranasal steroids. Four presented with Cushing's syndrome, three with growth failure, while two asymptomatic patients were discovered in the course of pituitary function testing. Four children had dysmorphic syndromes--Down's, Treacher-Collins, CHARGE association, and campomelic dysplasia--reflecting the vulnerability of such children to ENT problems, together with the difficulty of interpreting steroid induced growth failure in this context. Adrenal suppression was seen not only with betamethasone but also with budesonide, beclomethasone and flunisolide nasal preparations. A careful enquiry as to the use of intranasal steroids should be routine in children presenting with unexplained growth failure or Cushing's syndrome. Particular vigilance/awareness is required in children with dysmorphic syndromes. PMID- 12089124 TI - Two administration methods for inhaled salbutamol in intubated patients. AB - AIMS: To compare serum concentrations and effects on respiratory mechanics and haemodynamics of salbutamol administered by small volume nebuliser (SVN) and metered dose inhaler (MDI) plus spacer. METHODS: Blinded, randomised, crossover study in 12 intubated infants and children (mean age 17.8 months) receiving inhaled salbutamol therapy. Subjects received salbutamol as 0.15 mg/kg by SVN and four puffs (400 microg) by MDI plus spacer at a four hour interval in random order. Passive respiratory mechanics were measured by a single breath/single occlusion technique, and serum salbutamol concentrations by liquid chromatography mass spectrometry at 30 minutes, 1, 2, and 4 hours after each dose. Haemodynamics (heart rate and blood pressure) were recorded at each measurement time. RESULTS: There was no difference in percentage change in respiratory mechanics or haemodynamics between the two methods of administration. Mean area under the curve (AUC(0-4)) was 5.86 for MDI plus spacer versus 4.93 ng/ml x h for SVN. CONCLUSIONS: Serum concentrations and effects on respiratory mechanics and haemodynamics of salbutamol were comparable with the two administration methods under the conditions studied. Future studies are needed to determine the most effective and safe combination of dose and administration method of inhaled salbutamol in mechanically ventilated infants and children. PMID- 12089125 TI - Natural history and risk factors in fulminant hepatic failure. AB - BACKGROUND: The natural history of fulminant hepatic failure (FHF) without liver transplantation is not well known. AIMS: To study the natural history and prognostic factors, especially the presence of ascites and spontaneous bacterial peritonitis (SBP), in children with FHF. METHODS: FHF was defined by the onset of encephalopathy within 12 weeks of onset of jaundice. From August 1997 to December 2000, 67 children (< or =12 years) were diagnosed with FHF. Their clinical features, investigations and outcome were noted. Viral markers A to E (IgM, anti HAV; IgM, anti-HEV, HBsAg, and anti-HCV) were determined by ELISA. SBP was defined by the presence of > or =250 neutrophils with or without a positive culture in ascitic fluid. RESULTS: Mean age of the children was 5.8 years with an almost equal sex distribution. Viral markers were positive in 63 (94%) cases: hepatitis A in 34 (54%), E in 17 (27%), A+E in seven (11%), and B in five (8%). Thirty one children presented with grade I or II encephalopathy and all recovered, whereas 17 of 36 children who had grade III or IV encephalopathy died. Ascites was detected (both clinically and ultrasonically) in 34 (51%) cases, nine (26%) of which had SBP. Overall mortality was 25%. Mortality was higher in those who had ascites than in those who did not (32% v 18%); among those with ascites it was maximum in those who had SBP (78% v 16%). Total serum bilirubin and grade of encephalopathy were significantly higher, serum albumin was significantly lower, and prothrombin time was significantly prolonged in those who died than in those who recovered. CONCLUSION: The natural history of FHF in Indian children depends on age, grade of encephalopathy, ascites, and SBP. SBP depicts worse outcome. In all cases of FHF with ascites, the presence of SBP should be investigated. PMID- 12089126 TI - Bone mineral density in patients with classic galactosaemia. AB - BACKGROUND: Diminished bone mineral density (BMD) is a well known complication in women with classic galactosaemia caused by premature ovarian failure. Diminished BMD in prepubertal patients of either sex has, however, only been reported once. AIM: To assess BMD in children with classic galactosaemia. METHODS: Eleven treated patients (five males, six females, aged 2-18 years) had BMD determined by dual energy x ray absorptiometry. Two measurements were performed, an areal measurement of the total body and a volumetric measurement of the femoral neck. Results were expressed as Z scores. Dietary calcium intake, blood calcium, phosphate, vitamin D, parathormone, and markers of bone formation (bone alkaline phosphatase, osteocalcin) and bone resorption (NTX) were determined. RESULTS: All patients had a significantly diminished BMD. Mean Z score of the volumetric BMD was -1.76 (range -0.7 to -3.3), and of the areal BMD -0.99 (range -0.5 to -1.4). Dietary calcium intake and calcium, phosphate, parathormone, bone alkaline phosphatase, vitamin D metabolites, and osteocalcin (free and carboxylated) were normal in all patients. NTX levels in blood were significantly lower (p < 0.001) than in control subjects. CONCLUSION: BMD in this group of children of both sexes with classic galactosaemia under dietary treatment was decreased. Lower NTX levels in galactosaemics point to an apparent decreased bone resorption. PMID- 12089127 TI - Congenital heart disease and neuroblastoma: just coincidence? AB - The clinical history of a neonate with simple transposition of the great arteries in whom a metastatic neuroblastoma was diagnosed incidentally at autopsy is described, and the literature containing all 66 previously reported cases of neuroblastoma associated with congenital cardiac malformations is reviewed. One third of the described cases were classified as in situ neuroblastoma; neural crest derived cardiac lesions were present in 31%. Several possible aetiological mechanisms are discussed, and we conclude that the association of neuroblastoma with congenital cardiac malformations is multifactorial in origin. The described case represents the first reported example in which catecholamine release may have contributed to the fatal outcome of definitive congenital cardiac surgery. PMID- 12089128 TI - Neurofibromatosis type 1 and sporadic optic gliomas. AB - AIMS: To compare the natural history of sporadic optic glioma with those associated with neurofibromatosis type 1 (NF1). METHODS: Optic glioma cases were identified using both the Manchester Children's Tumour Registry (CTR) and the North West Regional NF1 Database (NF1DB), with detailed information on natural history available from the former (in 34 of 36 cases identified). RESULTS: A total of 52 cases over a period of 41 years were identified. From the 34 whose natural history was known, almost all (n = 31) were symptomatic, with mean ages of presentation of 4.5 and 5.1 years for NF1 and sporadic cases respectively. The majority (n = 22) presented with visual impairment, seven of whom were blind in at least one eye. Sporadic cases were over twice as likely as NF1 to have visual impairment. Recurrence occurred in 12 patients. Fewer NF1 patients died as a direct result of their optic glioma, but overall mortality and 5 and 10 year survival rates between the two groups were similar. All five primary (non metastatic) second central nervous system (CNS) tumours occurred in NF1 cases, two of these following radiotherapy. CONCLUSIONS: Symptomatic sporadic optic gliomas presented with impaired vision more frequently and were more aggressive than NF1 optic gliomas. Only optic glioma cases with NF1 were at risk of developing a second CNS tumour. Aggressive treatment of sporadic optic gliomas and early surveillance of NF1 optic gliomas may be required. The use of radiotherapy in these children requires further clarification. PMID- 12089130 TI - Intravenous atropine treatment in infantile hypertrophic pyloric stenosis. AB - AIMS: To assess the efficacy of a new regimen of intravenous atropine treatment for infantile hypertrophic pyloric stenosis (IHPS) with special reference to regression of pyloric hypertrophy. METHODS: Atropine was given intravenously at a dose of 0.01 mg/kg six times a day before feeding in 19 patients with IHPS diagnosed from radiographic and ultrasonographic findings. When vomiting ceased and the infants were able to ingest 150 ml/kg/day formula after stepwise increases in feeding volume, they were given 0.02 mg/kg atropine six times a day orally and the dose was decreased stepwise. RESULTS: Of the 19 infants, 17 (89%) ceased projectile vomiting after treatment with intravenous (median seven days) and subsequent oral (median 44 days) atropine administration. The remaining two infants required surgery. No significant complications were encountered. Ultrasonography showed a significant (p < 0.05) decrease in pyloric muscle thickness, but no significant shortening of the pyloric canal after completion of the atropine treatment. The patients exhibited failure to thrive at presentation, but were thriving at 6 months of age (p < 0.01). CONCLUSIONS: This atropine therapy resulted in satisfactory clinical recovery. Pyloric muscle thickness was significantly reduced. PMID- 12089131 TI - Nature and nurture in vitamin B12 deficiency. AB - We report on a child in whom severe nutritional vitamin B12 deficiency was exacerbated by a genetic impairment of the folate cycle, causing reduced CSF concentrations of the methyl group donor 5-methyltetrahydrofolate. Some patients with vitamin B12 deficiency may benefit from high dose folic acid supplementation, even if plasma concentrations are high. PMID- 12089132 TI - Does iron have a place in the management of breath holding spells? PMID- 12089134 TI - Is omeprazole helpful in the management of children with reflux oesophagitis? PMID- 12089135 TI - Does oral sucrose reduce the pain of neonatal procedures? PMID- 12089136 TI - Mentoring clinical researchers. PMID- 12089138 TI - C difficile induced pneumatosis intestinalis in a neutropenic child. PMID- 12089139 TI - Broad requirement for the mediator subunit RGR-1 for transcription in the Caenorhabditis elegans embryo. AB - The Mediator-related transcription co-factors integrate positive and negative inputs and recruit and activate the RNA polymerase II complex. To understand the role of Mediator during transcription, it is important to identify Mediator subunits that are essential for its functions. In the yeast Mediator, the conserved component Rgr1 is associated with multiple subunits that are required for specific activation or repression events. Yeast rgr1 is essential for viability, for certain repression mechanisms, and for activation of heat shock genes, but it is not known whether rgr1 is generally important for transcription. Here we have performed the first analysis of rgr-1 function in a metazoan. We found that in the developing Caenorhabditis elegans embryo rgr-1 is broadly required for transcription and for phosphorylation of both Ser-2 and Ser-5 of the RNA polymerase II C-terminal domain repeat. We conclude that RGR-1 fulfills a critical Mediator function that is broadly essential for metazoan mRNA transcription and that RGR-1 may be required at an early recruitment or initiation step. PMID- 12089140 TI - A third bacterial system for the assembly of iron-sulfur clusters with homologs in archaea and plastids. AB - The assembly of iron-sulfur (Fe-S) clusters is mediated by complex machinery. In several proteobacteria, this process involves ISC (Fe-S cluster assembly) machinery composed of at least six components also conserved in mitochondria from lower to higher eukaryotes. In nitrogen-fixing bacteria, another system, termed NIF (nitrogen fixation), is required for the maturation of nitrogenase. Here we report the identification of a third system, designated the SUF machinery, the components of which are encoded in Escherichia coli by an unassigned operon, sufABCDSE. We have analyzed spontaneous pseudorevertants isolated from a mutant strain lacking all the components of the ISC machinery. The suppressor mutations in the revertants have been localized to the regulatory region of the suf operon; overexpression of this operon restores the growth phenotypes and activity of Fe-S proteins in mutant cells lacking ISC. Disruption of the suf operon alone does not cause any major defects, but synthetic lethality was observed when both the isc and suf operons were inactivated. These results indicate that proteins encoded by the suf operon participate in the ISC-independent minor pathway for the assembly of Fe-S clusters. The genes homologous to sufBC are present in a wide range of bacteria, Archaea, and plastids, suggesting that this type of system is almost ubiquitous in nature. PMID- 12089141 TI - Detoxification of cholera toxin without removal of its immunoadjuvanticity by the addition of (STa-related) peptides to the catalytic subunit. A potential new strategy to generate immunostimulants for vaccination. AB - Peptides related to the heat-stable enterotoxin STa were fused to the N terminus of the A-subunit of cholera toxin (CTA) to explore whether peptide additions could help generate detoxified cholera toxin (CT) derivatives. Proteins carrying APRPGP (6-CTA), ASRCAELCCNPACPAP (16-CTA), or ANSSNYCCELCCNPACTGCYPGP (23-CTA) were genetically constructed. Using a two-plasmid system these derivatives were co-expressed in Vibrio cholerae with cholera toxin B-subunit (CTB) to allow formation and secretion of holotoxin-like molecules (engineered CT, eCTs). Purified eCTs maintained all normal CT properties yet they were more than 10-fold (eCT-6), 100-fold (eCT-16), or 1000-fold (eCT-23) less enterotoxic than wild-type CT. The inverse correlation between enterotoxicity and peptide length indicated sterical interference with the ADP-ribosylating active site in CTA. This interpretation agreed with greater than 1000-fold reductions in cAMP induction, with reductions, albeit not proportional, in in vitro agmatine ADP-ribosylation, and was supported by molecular simulations. Intranasal immunization of mice demonstrated that eCTs retained their inherent immunogenicity and ability to potentiate immune responses to a co-administered heterologous protein antigen, although in variable degrees. Therefore, the addition of STa-related peptides to CTA reduced the toxicity of CT while partly preserving its natural immunoadjuvanticity. These results suggest peptide extensions to CTA are a useful alternative to site-directed mutagenesis to detoxify CT. The simplicity of the procedure, combined with efficient expression and assembly of derivatives, suggests this approach could allow for large scale production of detoxified, yet immunologically active CT molecules. PMID- 12089142 TI - Involvement of toll-like receptor (TLR) 2 and TLR4 in cell activation by mannuronic acid polymers. AB - The alginate capsule produced by the human pathogen Pseudomonas aeruginosa is composed mainly of mannuronic acid polymers (poly-M) that have immunostimulating properties. Poly-M shares with lipopolysaccharide the ability to stimulate cytokine production from human monocytes in a CD14-dependent manner. In the present study we examined the role of Toll-like receptor (TLR) 2 and TLR4 in responses to poly-M. Blocking antibodies to TLR2 and TLR4 partly inhibited tumor necrosis factor production induced by poly-M in human monocytes, and further inhibition was obtained by combining the antibodies. By transiently transfecting HEK293 cells, we found that membrane CD14 together with either TLR2 or TLR4/MD-2 could mediate activation by poly-M. Transfection of HEK293 cells with TLR2 and fluorescently labeled TLR4 followed by co-patching of TLR2 with an antibody revealed no association of these molecules on the plasma membrane. However, macrophages from the Tlr4 mutant C3H/HeJ mice and TLR4 knockout mice were completely non-responsive to poly-M, whereas the tumor necrosis factor release from TLR2 knockout macrophages was half of that seen with wild type cells. Taken together the results suggest that both TLR2 and TLR4 are involved in cell activation by poly-M and that TLR4 may be required in primary murine macrophages. PMID- 12089144 TI - Constitutive agonist-independent CCR5 oligomerization and antibody-mediated clustering occurring at physiological levels of receptors. AB - Although homo-oligomerization has been reported for several G protein-coupled receptors, this phenomenon was not studied at low concentrations of receptors. Furthermore, it is not clear whether homo-oligomerization corresponds to an intrinsic property of nascent receptors or if it is a consequence of receptor activation. Here CCR5 receptor oligomerization was studied by bioluminescence resonance energy transfer (BRET) in cells expressing physiological levels of receptors. A strong energy transfer could be observed, in the absence of ligands, in whole cells and in both endoplasmic reticulum and plasma membrane subfractions, supporting the hypothesis of a constitutive oligomerization that occurs early after biosynthesis. No change in BRET was observed upon agonist binding, indicating that the extent of oligomerization is unrelated to the activation state of the receptor. In contrast, a robust increase of BRET, induced by a monoclonal antibody known to promote receptor clustering, suggests that microaggregation of preformed receptor homo-oligomers can occur. Taken together, our data indicate that constitutive receptor homo-oligomerization has a biologically relevant significance and might be involved in the process of receptor biosynthesis. PMID- 12089143 TI - cAMP inhibition of Akt is mediated by activated and phosphorylated Rap1b. AB - Rap1b has been implicated in the transduction of the cAMP mitogenic signal. Rap1b is phosphorylated and activated by cAMP, and its expression in cells where cAMP is mitogenic leads to an increase in G(1)/S phase entry and tumor formation. The PCCL3 thyroid follicular cells represent a differentiated and physiologically relevant system that requires thyrotropin (TSH), acting via cAMP, for a full mitogenic response. In this model system, cAMP stimulation of DNA synthesis requires activation and phosphorylation of Rap1b by the cAMP-dependent protein kinase A (PKA). This scenario presents the challenge of identifying biochemical processes involved in the phosphorylation-dependent Rap1b mitogenic action. In thyroid cells, Akt has been implicated in the stimulation of cell proliferation by TSH and cAMP. However, the mechanism(s) by which cAMP regulates Akt activity remains unclear. In this study we show that in PCCL3 cells 1) TSH inhibits Akt activity via cAMP and PKA; 2) Rap1b is required for cAMP inhibition of Akt; and 3) transduction of the cAMP signal into Akt requires activation as well as phosphorylation of Rap1b by PKA. PMID- 12089145 TI - Identification of calcium-independent phospholipase A2 (iPLA2) beta, and not iPLA2gamma, as the mediator of arginine vasopressin-induced arachidonic acid release in A-10 smooth muscle cells. Enantioselective mechanism-based discrimination of mammalian iPLA2s. AB - The agonist-stimulated release of arachidonic acid (AA) from cellular phospholipids in many cell types (e.g. myocytes, beta-cells, and neurons) has been demonstrated to be primarily mediated by calcium-independent phospholipases A(2) (iPLA(2)s) that are inhibited by the mechanism-based inhibitor (E)-6 (bromomethylene)-3-(1-naphthalenyl)-2H-tetrahydropyran-2-one (BEL). Recently, the family of mammalian iPLA(2)s has been extended to include iPLA(2)gamma, which previously could not be pharmacologically distinguished from iPLA(2)beta. To determine whether iPLA(2)beta or iPLA(2)gamma (or both) were the enzymes responsible for arginine vasopressin (AVP)-induced AA release from A-10 cells, it became necessary to inhibit selectively iPLA(2)beta and iPLA(2)gamma in intact cells. We hypothesized that the R- and S-enantiomers of BEL would possess different inhibitory potencies for iPLA(2)beta and iPLA(2)gamma. Accordingly, racemic BEL was separated into its enantiomeric constituents by chiral high pressure liquid chromatography. Remarkably, (S)-BEL was approximately an order of magnitude more selective for iPLA(2)beta in comparison to iPLA(2)gamma. Conversely, (R)-BEL was approximately an order of magnitude more selective for iPLA(2)gamma than iPLA(2)beta. The AVP-induced liberation of AA from A-10 cells was selectively inhibited by (S)-BEL (IC(50) approximately 2 microm) but not (R) BEL, demonstrating that the overwhelming majority of AA release is because of iPLA(2)beta and not iPLA(2)gamma activity. Furthermore, pretreatment of A-10 cells with (S)-BEL did not prevent AVP-induced MAPK phosphorylation or protein kinase C translocation. Finally, two different cell-permeable protein kinase C activators (phorbol-12-myristate-13-acetate and 1,2-dioctanoyl-sn-glycerol) could not restore the ability of A-10 cells to release AA after exposure to (S)-BEL, thus supporting the downstream role of iPLA(2)beta in AVP-induced AA release. PMID- 12089146 TI - Modular arrangement and secretion of a multidomain serine protease. Evidence for involvement of proline-rich region and N-glycans in the secretion pathway. AB - The Limulus Factor C (FC), a multidomain glycoprotein that binds bacterial endotoxin with high affinity, belongs to the serine protease family of the complement and blood coagulation cascade. Here, we provide compelling evidence for the importance of modular arrangement and relevance of the proline-rich region (PRR) and N-glycosylation to the secretion and function of FC. We propose that PRR could be a universal conformational domain that regulates protein folding and targeting. FCs lacking PRR preceding the serine protease domain, were localized intracellularly. Misfolded conformers of the intracellular FCs were more susceptible to trypsin digestion. Glycosylation inhibition studies indicate that the presence but not the exact structure of the N-glycans affects the secretion of FC, although the complexity of glycosylation may influence its endotoxin-induced proteolytic cleavage with resultant enzymatic activity. Disruption of specific N-glycan sites at positions 740, 767, and 912, downstream of the PRR, at or near the serine protease domain, blocks its secretion. Co expressed molecular chaperones like canine calnexin associates with glycosylated FCs to increase its solubility and secretion level but did not alter their expression profiles. Our results clearly demonstrate that the folding and secretion of a multidomain serine protease like FC are determined by its modular domain arrangement and site-specific N-glycans. The secreted FCs containing the N terminal portion of FC are able to detect lipopolysaccharide with high sensitivity. We also identified the lectin-like and sushi 4 domains to contribute to the binding of lipopolysaccharide. PMID- 12089147 TI - Calmodulin activates electron transfer through neuronal nitric-oxide synthase reductase domain by releasing an NADPH-dependent conformational lock. AB - Neuronal nitric-oxide synthase (nNOS) is activated by the Ca(2+)-dependent binding of calmodulin (CaM) to a characteristic polypeptide linker connecting the oxygenase and reductase domains. Calmodulin binding also activates the reductase domain of the enzyme, increasing the rate of reduction of external electron acceptors such as cytochrome c. Several unusual structural features appear to control this activation mechanism, including an autoinhibitory loop, a C-terminal extension, and kinase-dependent phosphorylation sites. Pre-steady state reduction and oxidation time courses for the nNOS reductase domain indicate that CaM binding triggers NADP(+) release, which may exert control over steady-state turnover. In addition, the second order rate constant for cytochrome c reduction in the absence of CaM was found to be highly dependent on the presence of NADPH. It appears that NADPH induces a conformational change in the nNOS reductase domain, restricting access to the FMN by external electron acceptors. CaM binding reverses this effect, causing a 30-fold increase in the second order rate constant. The results show a startling interplay between the two ligands, which both exert control over the conformation of the domain to influence its electron transfer properties. In the full-length enzyme, NADPH binding will probably close the conformational lock in vivo, preventing electron transfer to the oxygenase domain and the resultant stimulation of nitric oxide synthesis. PMID- 12089148 TI - Low intracellular zinc impairs the translocation of activated NF-kappa B to the nuclei in human neuroblastoma IMR-32 cells. AB - In the current work, we studied how variations in extracellular zinc concentrations modulate different steps involved in nuclear factor kappaB (NF kappaB) activation in human neuroblastoma IMR-32 cells. Cells were incubated in media containing varying concentrations of zinc (1.5, 5, 15, and 50 microm). Within 3 h, the intracellular zinc content was lower in cells exposed to 1.5 and 5 microm, compared with the other groups. Low intracellular zinc concentrations were associated with the activation of NF-kappaB, based on high levels of IkappaBalpha phosphorylation, low IkappaBalpha concentrations, and high NF-kappaB binding activity in total cell fractions. However, the active dimer accumulated in the cytosol, as shown by a low ratio of nuclear/cytosolic NF-kappaB binding activity. This altered nuclear translocation was accompanied by a decreased transactivation of an endogenous NF-kappaB-driven gene (ikba) and of a reporter gene (pNF-kappaB-luc). In cells with low intracellular zinc concentrations, a low rate of in vitro tubulin polymerization was measured compared with the other groups. We conclude that low intracellular zinc concentrations induce tubulin depolymerization, which may be one signal for NF-kappaB activation. However, NF kappaB nuclear translocation is impaired, which inhibits the transactivation of NF-kappaB-driven genes. This could affect cell survival, and be an important factor in certain zinc deficiency-associated pathologies. PMID- 12089149 TI - Molecular identification of a novel carnitine transporter specific to human testis. Insights into the mechanism of carnitine recognition. AB - l-Carnitine is an essential component of mitochondrial fatty acid beta-oxidation and plays a pivotal role in the maturation of spermatozoa within the male reproductive tract. Epididymal plasma contains the highest levels of l-carnitine found in the human body, and initiation of sperm motility occurs in parallel to l carnitine increase in the epididymal lumen. Using a specific carrier, epididymal epithelium secretes l-carnitine into the lumen by an active transport mechanism; however, the structure-activity relationship comprising the carnitine-permeation pathway is poorly understood. We discovered a novel carnitine transporter (CT2) specifically located in human testis. Analyzing the primary structure of CT2 revealed that it is phylogenetically located between the organic cation transporter (OCT/OCTN) and anion transporter (OAT) families. Hence, CT2 represents a novel transporter family. When expressed in Xenopus oocytes, CT2 mediates the high affinity transport of l-carnitine but does not accept mainstream OCT/OCTN cationic or OAT anionic substrates. We synthesized and tested various carnitine-related compounds and investigated the physicochemical properties of substrate recognition by semi-empirical computational chemistry. The data suggest that the quaternary ammonium cation bulkiness and relative hydrophobicity be the most important factors that trigger CT2-substrate interactions. Immunohistochemistry showed that the CT2 protein is located in the luminal membrane of epididymal epithelium and within the Sertoli cells of the testis. The identification of CT2 represents an interesting evolutionary link between OCT/OCTNs and OATs, as well as provides us with an important insight into the maturation of human spermatozoa. PMID- 12089150 TI - Exposure of yeast cells to anoxia induces transient oxidative stress. Implications for the induction of hypoxic genes. AB - The mitochondrial respiratory chain is required for the induction of some yeast hypoxic nuclear genes. Because the respiratory chain produces reactive oxygen species (ROS), which can mediate intracellular signal cascades, we addressed the possibility that ROS are involved in hypoxic gene induction. Recent studies with mammalian cells have produced conflicting results concerning this question. These studies have relied almost exclusively on fluorescent dyes to measure ROS levels. Insofar as ROS are very reactive and inherently unstable, a more reliable method for measuring changes in their intracellular levels is to measure their damage (e.g. the accumulation of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in DNA, and oxidative protein carbonylation) or to measure the expression of an oxidative stress-induced gene, e.g. SOD1. Here we used these approaches as well as a fluorescent dye, carboxy-H(2)-dichloro-dihydrofluorescein diacetate (carboxy-H(2) DCFDA), to determine whether ROS levels change in yeast cells exposed to anoxia. These studies reveal that the level of mitochondrial and cytosolic protein carbonylation, the level of 8-OH-dG in mitochondrial and nuclear DNA, and the expression of SOD1 all increase transiently during a shift to anoxia. These studies also reveal that carboxy-H(2)-DCFDA is an unreliable reporter of ROS levels in yeast cells shifted to anoxia. By using two-dimensional electrophoresis and mass spectrometry (matrix-assisted laser desorption ionization time-of flight), we have found that specific proteins become carbonylated during a shift to anoxia and that some of these proteins are the same proteins that become carbonylated during peroxidative stress. The mitochondrial respiratory chain is responsible for much of this carbonylation. Together, these findings indicate that yeast cells exposed to anoxia experience transient oxidative stress and raise the possibility that this initiates the induction of hypoxic genes. PMID- 12089151 TI - A novel family 8 xylanase, functional and physicochemical characterization. AB - Xylanases are generally classified into glycosyl hydrolase families 10 and 11 and are found to frequently have an inverse relationship between their pI and molecular mass values. However, we have isolated a psychrophilic xylanase that belongs to family 8 and which has both a high pI and high molecular mass. This novel xylanase, isolated from the Antarctic bacterium Pseudoalteromonas haloplanktis, is not homologous to family 10 or 11 enzymes but has 20-30% identity with family 8 members. NMR analysis shows that this enzyme hydrolyzes with inversion of anomeric configuration, in contrast to other known xylanases which are retaining. No cellulase, chitosanase or lichenase activity was detected. It appears to be functionally similar to family 11 xylanases. It hydrolyzes xylan to principally xylotriose and xylotetraose and is most active on long chain xylo-oligosaccharides. Kinetic studies indicate that it has a large substrate binding cleft, containing at least six xylose-binding subsites. Typical psychrophilic characteristics of a high catalytic activity at low temperatures and low thermal stability are observed. An evolutionary tree of family 8 enzymes revealed the presence of six distinct clusters. Indeed classification in family 8 would suggest an (alpha/alpha)(6) fold, distinct from that of other currently known xylanases. PMID- 12089152 TI - High resolution crystal structures of the wild type and Cys-55-->Ser and Cys-59- >Ser variants of the thioredoxin-like [2Fe-2S] ferredoxin from Aquifex aeolicus. AB - The [2Fe-2S] ferredoxin (Fd4) from Aquifex aeolicus adopts a thioredoxin-like polypeptide fold that is distinct from other [2Fe-2S] ferredoxins. Crystal structures of the Cys-55 --> Ser (C55S) and Cys-59 --> Ser (C59S) variants of this protein have been determined to 1.25 A and 1.05 A resolution, respectively, whereas the resolution of the wild type (WT) has been extended to 1.5 A. The improved WT structure provides a detailed description of the [2Fe-2S] cluster, including two features that have not been noted previously in any [2Fe-2S] cluster-containing protein, namely, pronounced distortions in the cysteine coordination to the cluster and a Calpha-H-Sgamma hydrogen bond between cluster ligands Cys-55 and Cys-9. These features may contribute to the unusual electronic and magnetic properties of the [2Fe-2S] clusters in WT and variants of this ferredoxin. The structures of the two variants of Fd4, in which single cysteine ligands to the [2Fe-2S] cluster are replaced by serine, establish the metric details of serine-ligated Fe-S active sites with unprecedented accuracy. Both the cluster and its surrounding protein matrix change in subtle ways to accommodate this ligand substitution, particularly in terms of distortions of the Fe(2)S(2) inorganic core from planarity and displacements of the polypeptide chain. These high resolution structures illustrate how the interactions between polypeptide chains and Fe-S active sites reflect combinations of flexibility and rigidity on the part of both partners; these themes are also evident in more complex systems, as exemplified by changes associated with serine ligation of the nitrogenase P cluster. PMID- 12089153 TI - Two E2F sites in the Arabidopsis MCM3 promoter have different roles in cell cycle activation and meristematic expression. AB - The commitment to DNA replication is a key step in cell division control. The Arabidopsis MCM3 homologue forms part of the mini chromosome maintenance (MCM) complex involved in the initiation of DNA replication at the transition G(1)/S. Consistent with its role at the G(1)/S transition we show that the AtMCM3 gene is transcriptionally regulated at S phase. The 5' region of this gene contains several E2F consensus binding sites, two of which match the human consensus closely and whose roles have been studied here. The identity of the two sequences as E2F binding sites has been confirmed by electrophoretic mobility shift assay analyses. Furthermore the promoter is activated by AtE2F-a and AtDP-a factors in transient expression studies. One of the E2F binding sites is shown to be responsible for the G(2)-specific repression of the promoter in synchronized cell suspension cultures. In contrast, the second E2F binding site has a role in meristem-specific expression in planta as deletion of this site eliminates the expression of a reporter gene in root and apical meristems. Thus two highly similar E2F binding sites in the promoter of the MCM3 gene are responsible for different cell cycle regulation or developmental expression patterns depending on the cellular environment. PMID- 12089154 TI - Fe65, a ligand of the Alzheimer's beta-amyloid precursor protein, blocks cell cycle progression by down-regulating thymidylate synthase expression. AB - The functions of the Alzheimer's beta-amyloid precursor protein (APP) and of its complex with the adaptor protein Fe65 are still unknown. We have demonstrated that Fe65 is also a nuclear protein and APP functions as an extranuclear anchor, thus preventing Fe65 nuclear translocation. According to this finding, it was also demonstrated that Fe65 could play a role in the regulation of transcription. In the present paper we show that the overexpression of Fe65 prevents G(1) --> S cell cycle progression of serum-stimulated fibroblasts and that the contemporary overexpression of APP abolishes this effect of Fe65. The overexpression of Fe65 completely abolishes the activation of a key S phase gene, the thymidylate synthase (TS) gene, driven by the transcription factor LSF/CP2/LBP1 (LSF). This phenomenon is observed only in experimental conditions leading to the accumulation of Fe65 in the nucleus. Similarly, the two other members of the Fe65 protein family, Fe65L1 and Fe65L2, have been found to translocate into the nucleus and to prevent the activation of the TS gene promoter induced by LSF. Two results support the hypothesis that the inhibitory effect of Fe65 on cell cycle progression in fibroblasts is the result of the inhibition of TS gene expression: (i) Fe65 overexpression, but not Fe65 and APP co-expression, prevents the accumulation of endogenous TS upon the exposure of cells to serum, and (ii) thymidine addition to the culture medium completely overcomes the growth arrest caused by Fe65. In neuronal PC12 cells, the overexpression of Fe65 or of Fe65L1 and Fe65L2 blocks cell cycle, as observed in fibroblasts, but thymidine supplementation to culture medium does not revert this block, thus suggesting that Fe65 proteins induce in neuronal cells a gene expression program different from that activated in fibroblasts. PMID- 12089155 TI - Botulinum neurotoxin A activity is dependent upon the presence of specific gangliosides in neuroblastoma cells expressing synaptotagmin I. AB - Botulinum neurotoxin A (BoNT/A) is the deadliest of all known biological substances. Although its toxicity makes BoNT/A a biological warfare threat, its biologic activity makes it an increasingly useful therapeutic agent for the treatment of muscular disorders. However, almost 200 years after its discovery, the neuronal cell components required for the activity of this deadly toxin have not been unequivocally identified. In this work, neuroblastoma cells expressing synaptotagmin I, a protein shown to be bound by BoNT/A, were used to determine whether specific gangliosides were necessary for BoNT/A activity as measured by synaptosomal-associated protein of 25 kDa (SNAP-25) cleavage. Ganglioside GT1b was found to support BoNT/A activity significantly more effectively than GD1a, which was far more effective than GM1 when added to ganglioside-deficient murine cholinergic Neuro 2a or to human adrenergic SK-N-SH neuroblastoma cells. Whereas both cell lines expressed synaptotagmin I, SNAP-25 cleavage was not observed in the absence of complex gangliosides. These results indicate that 1) gangliosides are required for BoNT/A activity, 2) synaptotagmin I in the absence of gangliosides does not support BoNT/A activity, and 3) Neuro 2a cells are an efficient model system for studying the biological activity of BoNT/A. PMID- 12089156 TI - A syndrome of joint laxity and impaired tendon integrity in lumican- and fibromodulin-deficient mice. AB - Lumican and fibromodulin regulate the assembly of collagens into higher order fibrils in connective tissues. Here, we show that mice deficient in both of these proteoglycans manifest several clinical features of Ehlers-Danlos syndrome. The Lum(-/-)Fmod(-/-) mice are smaller than their wild type littermates and display gait abnormality, joint laxity, and age-dependent osteoarthritis. Misaligned knee patella, severe knee dysmorphogenesis, and extreme tendon weakness are the likely causes for joint laxity in the double-nulls. Fibromodulin deficiency alone leads to significant reduction in tendon stiffness in the Lum(+/+)Fmod(-/-) mice, with further loss in stiffness in a Lum gene dose-dependent way. At the protein level, we show marked increase of lumican in Fmod(-/-) tendons, which may partially rescue the tendon phenotype in this genotype. These results establish fibromodulin as a key regulator and lumican as a modulator of tendon strength. A disproportionate increase in small diameter immature collagen fibrils and a lack of progression to mature, large diameter fibrils in the Fmod(-/-) background may constitute the underlying cause of tendon weakness and suggest that fibromodulin aids fibril maturation. This study demonstrates that the collagen fibril modifying proteoglycans, lumican and fibromodulin, are candidate genes and key players in the pathogenesis of certain types of Ehlers-Danlos syndrome and other connective tissue disorders. PMID- 12089158 TI - Molecular, biochemical, and functional characterization of a Nudix hydrolase protein that stimulates the activity of a nicotinoprotein alcohol dehydrogenase. AB - The cytoplasmic coenzyme NAD(+)-dependent alcohol (methanol) dehydrogenase (MDH) employed by Bacillus methanolicus during growth on C(1)-C(4) primary alcohols is a decameric protein with 1 Zn(2+)-ion and 1-2 Mg(2+)-ions plus a tightly bound NAD(H) cofactor per subunit (a nicotinoprotein). Mg(2+)-ions are essential for binding of NAD(H) cofactor in MDH protein expressed in Escherichia coli. The low coenzyme NAD(+)-dependent activity of MDH with C(1)-C(4) primary alcohols is strongly stimulated by a second B. methanolicus protein (ACT), provided that MDH contains NAD(H) cofactor and Mg(2+)-ions are present in the assay mixture. Characterization of the act gene revealed the presence of the highly conserved amino acid sequence motif typical of Nudix hydrolase proteins in the deduced ACT amino acid sequence. The act gene was successfully expressed in E. coli allowing purification and characterization of active ACT protein. MDH activation by ACT involved hydrolytic removal of the nicotinamide mononucleotide NMN(H) moiety of the NAD(H) cofactor of MDH, changing its Ping-Pong type of reaction mechanism into a ternary complex reaction mechanism. Increased cellular NADH/NAD(+) ratios may reduce the ACT-mediated activation of MDH, thus preventing accumulation of toxic aldehydes. This represents a novel mechanism for alcohol dehydrogenase activity regulation. PMID- 12089157 TI - Lipopolysaccharide down-regulates Sp1 binding activity by promoting Sp1 protein dephosphorylation and degradation. AB - We examined the in vivo effect of lipopolysaccharide (LPS) on Sp1 (promoter selective transcription factor 1) DNA binding activity and studied the mechanisms involved in mouse lungs. The Sp1 DNA complex displayed a major band composed of Sp1, Sp2, and Sp3 trimer and a minor band composed of Sp3 homodimer. Compared with control, nuclear proteins from lungs challenged with LPS for 60, 90, 120, 150, 180, and 240 min, respectively, showed a markedly reduced Sp1 binding activity. Down-regulation of Sp1 binding activity was accompanied by a reduced expression of two Sp1-dependent genes (endothelial nitric oxide synthase and cyclooxygenase-1). Immunoprecipitation-Western blot experiments demonstrated that LPS dephosphorylated Sp1 protein at serine and threonine residues but not at the tyrosine residue. Dephosphorylation of Sp1 protein in vitro significantly reduced Sp1 DNA binding activity. Deglycosylation of Sp1 protein also reduced Sp1 binding activity. However, LPS did not cause Sp1 deglycosylation. LPS markedly reduced nuclear Sp1 protein level but had no significant effect on Sp1 mRNA abundance and on Sp1 protein nuclear translocation. Both Sp1 protein dephosphorylation and Sp1 protein degradation are temporally correlated to the reduced Sp1 binding activity. Our results demonstrate that challenge of mice with LPS in vivo down regulates Sp1 DNA binding activity through promoting Sp1 protein dephosphorylation and degradation. PMID- 12089159 TI - H2, the minor subunit of the human asialoglycoprotein receptor, trafficks intracellularly and forms homo-oligomers, but does not bind asialo-orosomucoid. AB - The functional human hepatic asialoglycoprotein receptor (ASGP-R) is a hetero oligomer composed of two subunits, designated H1 and H2, which are highly homologous. Despite their extensive homology, the major H1 subunit is stably expressed by itself, whereas in the absence of H1 most of the H2 subunits are degraded in the ER. In this study, we were able to investigate the capability of the minor ASGP-R subunit, H2, to function independently of H1, because it was apparently stabilized by fusing its NH(2) terminus with an epitope tag. We could thus create stable cell lines in hepatoma-derived SK-Hep-1 cells that expressed the H2 subunit alone. H2 was expressed on the cell surface and was internalized, predominantly through the clathrin-coated pit pathway. Since the internal pool of H2 was also able to traffick to the cell surface, we conclude that H2 recycles between the surface and intracellular compartments, similar to the constitutive recycling of hetero-oligomeric ASGP-R complexes. However, the rate of H2 recycling and internalization was approximately 25-33% that of H1. Similar to H1, the H2 polypeptides were also able to self-associate to form homo-oligomers, including trimers and tetramers. However, unlike H1, which can bind the ligand asialo-orosomucoid (ASOR) when overexpressed in COS-7 cells, H2 failed to bind or endocytose ASOR. In summary, the H2 subunit of the human ASGP-R contains functional, although weak, signal(s) for endocytosis and recycling and has the ability to oligomerize. H2 homo-oligomers, however, do not create binding sites for desialylated glycoproteins, such as ASOR, that contain tri- and tetra antennary N-linked oligosaccharides. Nonetheless, these results raise the intriguing possibility that naturally occurring H2 homo-oligomers may exist in human hepatocytes and have an as yet undiscovered function. PMID- 12089160 TI - Active nuclear import and export pathways regulate E2F-5 subcellular localization. AB - Epidermal keratinocyte differentiation is accompanied by differential regulation of E2F genes, including up-regulation of E2F-5 and its concomitant association with the retinoblastoma family protein p130. This complex appears to play a role in irreversible withdrawal from the cell cycle in differentiating keratinocytes. We now report that keratinocyte differentiation is also accompanied by changes in E2F-5 subcellular localization, from the cytoplasm to the nucleus. To define the molecular determinants of E2F-5 nuclear import, we tested its ability to enter the nucleus in import assays in vitro using digitonin-permeabilized cells. We found that E2F-5 enters the nucleus through mediated transport processes that involve formation of nuclear pore complexes. It has been proposed that E2F-4 and E2F-5, which lack defined nuclear localization signal (NLS) consensus sequences, enter the nucleus in association with NLS-containing DP-2 or pRB family proteins. However, we show that nuclear import of E2F-5 only requires the first N-terminal 56 amino acid residues and is not dependent on interaction with DP or pRB family proteins. Because E2F-5 is predominantly cytoplasmic in undifferentiated keratinocytes and in other intact cells, we also examined whether this protein is subjected to active nuclear export. Indeed, E2F-5 is exported from the nucleus through leptomycin B-sensitive, CRM1-mediated transport, through a region corresponding to amino acid residues 130-154. This region excludes the DNA- and the p130-binding domains. Thus, the subcellular distribution of E2F-5 is tightly regulated in intact cells, through multiple functional domains that direct nucleocytoplasmic shuttling of this protein. PMID- 12089161 TI - Role of cardiac natriuretic peptide testing in heart failure. PMID- 12089162 TI - Improving results from home blood-glucose monitoring: accuracy and reliability require greater patient education as well as improved technology. PMID- 12089163 TI - A hematologic "gold standard" for iron-deficient states? PMID- 12089164 TI - Single-step assays to analyze CYP2D6 gene polymorphisms in Asians: allele frequencies and a novel *14B allele in mainland Chinese. AB - BACKGROUND: Cytochrome P450-dependent monooxygenase 2D6 (CYP2D6) activity can be estimated by investigating the metabolism of model drugs or by genotyping the most common CYP2D6 alleles. For Caucasians, the CYP2D6 allele frequencies are well investigated, and single-step assays are available for genotyping, whereas allele analysis in mainland Chinese is limited. METHODS: Two tetra-primer assays and one allele-specific amplification assay were developed to easily genotype the CYP2D6 alleles *8, *10, and *14 previously detected in Asians. Applying these assays in combination with established single-tube assays, we analyzed 223 DNA samples from Chinese volunteers for the CYP2D6 alleles *3, *4, *5, *6, *8, *10, and *14 and for duplication of CYP2D6. RESULTS: Six different alleles were detected in mainland Chinese. The most frequent mutant allele was the intermediate metabolizer allele, CYP2D6*10, with a prevalence of 51.3%, followed by the poor metabolizer alleles CYP2D6*5 (7.2%) and a novel variant of CYP2D6*14. This novel *14B allele (2.0%) differs from the *14 allele by the absence of the C188T substitution and by the additional G1749C substitution. Furthermore, six duplication alleles of CYP2D6 were detected, including one duplication of the *10 allele (*10X2). CONCLUSIONS: The CYP2D6 allele frequencies in mainland Chinese shows some genetic diversity compared with Chinese from other regions: a novel *14B allele, a slightly higher frequency of the *5 allele, and a slightly lower frequency of the *10 allele than in most other Chinese populations. PMID- 12089165 TI - Independent risk factor for moderate to severe internal carotid artery stenosis: T786C mutation of the endothelial nitric oxide synthase gene. AB - BACKGROUND: NO synthesized from L-arginine by the constitutive endothelial NO synthase (eNOS) plays a key role in the atherosclerotic process. We investigated whether common variants in the NOS3 gene (a T786C mutation in the 5' flanking region and the polymorphism on exon 7 that produced the Glu298Arg polymorphism in the protein) are associated with an increased risk of moderate to severe internal carotid artery (ICA) stenosis. METHODS: We studied 88 patients consecutively operated for ICA stenosis and 133 healthy controls. A T786C mutation in the 5' flanking region and the polymorphism in exon 7 that produces the Glu298Asp polymorphism in the protein were explored by PCR and fluorescent probe analysis. RESULTS: Genotype distribution was significantly different between patients and controls only for T786C, the CC genotype frequency being 26% and 13%, respectively [odds ratio (OR), 2.26; 95% confidence interval (CI), 1.14-4.46; P = 0.018]. Moreover, the CC genotype was significantly more frequent in a subgroup of patients with ulcerative plaques compared with patients with nonulcerative lesions (44% vs 17%; OR, 3.82; 95% CI, 1.79-8.14; P = 0.003). Multiple logistic regression analysis using the most frequent risk factors and the eNOS gene variant showed that the CC genotype is an independent risk factor for ICA stenosis (P = 0.023). CONCLUSION: C allele homozygosity in position 786 of the eNOS promoter seems to be an independent risk factor for the development of moderate to severe ICA stenosis, especially ulcerative lesions. PMID- 12089166 TI - Instruments for self-monitoring of blood glucose: comparisons of testing quality achieved by patients and a technician. AB - BACKGROUND: Instruments for self-monitoring of blood glucose (SMBG) are increasingly used by patients with diabetes. The analytical quality of meters in routine use is poorly characterized. METHODS: We compared SMBG performance achieved by patients and by a medical laboratory technician. Imprecision was calculated from duplicate measurements, and deviation as the difference between the first measurement and the mean of duplicate laboratory-method results (calibrated with NIST material). Analytical quality for five groups of SMBG instruments was compared with quality specifications for BG measurements. All participants completed a questionnaire assessing both SMBG training and use of the meters. RESULTS: We recruited 159 SMBG users from a hospital outpatient clinic and 263 others from 65 randomly selected general practices (total of 422). Most (two thirds) used insulin. CVs for the five meter types were 7%, 11%, 18%, 18%, and 20% in the hands of patients and 2.5-5.9% for the technician. For three of five meter types, patients' BG measurements had larger deviations from the laboratory results than did the technician's results. The technician's performance could not predict the patients'. No instrument when used by patients (but two operated by the technician) met published quality specifications. The analytical quality of patients' results was not related to whether they had chosen the instruments on advice from healthcare personnel (one-third of patients), were only self-educated in SMBG (50%), or performed SMBG fewer than seven times/week (62%). CONCLUSIONS: The analytical quality of SMBG among patients was poorer than, and could not be predicted from, the performance of the meters in the hands of a technician. We suggest that new instruments be tested in the hands of patients who are trained on meter use in a routine way. PMID- 12089167 TI - Phospholipid autoantibodies and the antiphospholipid antibody syndrome: diagnostic accuracy of 23 methods studied by variation in ROC curves with number of clinical manifestations. AB - BACKGROUND: We analyzed the diagnostic accuracies for the diagnosis of antiphospholipid syndrome (APS) of 23 antiphospholipid antibody (APL-Ab) assays targeted at different antigen preparations and immunoglobulin isotypes. METHODS: In 144 patients with suspected APS, anti-cardiolipin (aCL) and anti-beta2 glycoprotein I (abeta2GPI) antibodies were measured with 23 different ELISAs from three manufacturers. Data were analyzed by ROC curves. In the absence of an accepted criterion standard, the endpoint "diagnosis of APS" was varied according to the number (two through five) of signs and symptoms of APS. RESULTS: Although the presence of lupus anticoagulant was associated significantly with APL-Ab in 10 of 23 assays (P = 0.01-10(-4)) and recurrent arterial or venous occlusions were significantly associated with APL-Ab of IgM isotype in 5 of 6 assays (P = 0.02-10(-4)), sensitivity for detection of APS did not exceed 67%. With the exception of IgA APL-Ab, the diagnostic accuracy of the assays improved when the diagnosis of APS was based on an increasing number of simultaneous features of APS. For most methods, areas under the ROC curves were >0.8 irrespective of the method's subclass specificity and antigen preparation (aCL or abeta2GPI), if the clinical diagnosis of APS was based on four or more signs and symptoms of APS. CONCLUSION: Despite considerable heterogeneity in the individual test results, a single test of IgG or IgM isotype targeted at either aCL or abeta2GPI antibodies has excellent diagnostic accuracy when the criterion for diagnosis requires four or more typical manifestations of APS. PMID- 12089168 TI - Long-term analytical performance of hemostasis field methods as assessed by evaluation of the results of an external quality assessment program for antithrombin. AB - BACKGROUND: It is important for a laboratory to know the stability of performance of laboratory tests over time. The aim of this study was to adapt from the field of clinical chemistry a method to assess the long-term analytical performance of hemostasis field methods. METHODS: The linear regression model was used to compare the laboratory results with the consensus mean value of a survey. This model was applied to plasma antithrombin activity using the data for 82 laboratories, collected between 1996 and 1999 in the European Concerted Action on Thrombosis (ECAT) external quality assessment program. The long-term total, random, and systematic error were calculated. The variables introduced to define the long-term performance in this model were the long-term analytical CV (LCV(a)) and the analytical critical difference (ACD), which indicates the minimum difference necessary between two samples measured on a long-term time-scale to consider them statistically significantly different. RESULTS: The systematic error (bias) ranged from 4.5 to 103 units/L. The random error ranged from 24.4 to 242 units/L. For the majority of the laboratories, random error was the main component (>75%) of the total error. The LCV(a), after adjustment for the contribution of the bias, ranged from 2.8% to 48%. The ACD ranged from 78 to 1290 units/L with a median value of 190 units/L. No statistically significant differences were observed for either LCV(a) or ACD between the two different measurement principles for antithrombin activity based on the inhibition of either thrombin or factor Xa. CONCLUSIONS: This linear regression model is useful for assessing the total error, random error, and bias for hemostasis field methods. The LCV(a) and ACD for measurement on a long-term time-scale appear to be useful for assessing the long-term analytical performance. PMID- 12089169 TI - Bioluminescent method for detecting telomerase activity. AB - BACKGROUND: Telomerase is a promising biomarker in cancer diagnosis and therapy. The elongation of telomeric repeats catalyzed by telomerase is accompanied by release of six PP(i) for each TTAGGG repeat (1 pmol PP(i)/310 pg telomeric repeats). We developed a novel method to measure telomerase activity by use of an enzymatic luminometric PP(i) assay (ELIPA). METHODS: Extracts of cell lines and tissues were incubated with primer at 30 degrees C for 30 min. Released PP(i) was converted to ATP by sulfurylase, and ATP was detected by a luciferase bioluminescence system. The ELIPA results were compared with results obtained with the conventional telomeric repeat amplification (TRAP)-ELISA in 42 lung carcinoma tissues and 27 control tissues without malignancy. RESULTS: The lower detection limits of ELIPA and TRAP-ELISA were 5 and 10 cells, respectively. The within-run imprecision (CV) of ELIPA was < or =12%. When compared with TRAP ELISA, the correlation coefficient (r) was 0.79. When we used the cutoff value from ROC analysis to distinguish malignant and nonmalignant tissues, the sensitivity and specificity of ELIPA were 83% and 96%, respectively, whereas the sensitivity and specificity of TRAP-ELISA were 71% and 96%, respectively. CONCLUSION: ELIPA is a simple and sensitive homogeneous method to quantify telomerase activity. PMID- 12089170 TI - Quantification and utility of monosialylated alpha-fetoprotein in the diagnosis of hepatocellular carcinoma with nondiagnostic serum total alpha-fetoprotein. AB - BACKGROUND: At concentrations <500 microg/L, serum alpha-fetoprotein (AFP) has low specificity in the diagnosis of hepatocellular carcinoma (HCC), but monosialylated AFP (msAFP) is more specific for HCC. We describe two strategies for quantitative analysis of msAFP and explore their diagnostic accuracy in cases of HCC with nondiagnostic serum total AFP concentrations. METHODS: We first used isoelectric focusing, Western blot, and densitometry (IEF-Western blot assay). We then developed a second assay, a novel glycosylation immunosorbent assay (GISA), based on the specificity of sialyltransferase and immunosorbent technology. Both assays were used to measure msAFP and msAFP percentage relative to total AFP in sera with nondiagnostic AFP concentrations from 36 patients with newly diagnosed HCC and from 18 patients with liver cirrhosis. RESULTS: The msAFP percentages and concentrations were significantly higher in the HCC patient group regardless of the quantification methods. The msAFP concentrations and msAFP percentages obtained by the two assays were highly correlated (r = 0.70 and 0.49, respectively). For discrimination of HCC with nondiagnostic serum total AFP from liver cirrhosis, the areas under the ROC curves were 0.81 (95% confidence interval, 0.70-0.92) for msAFP by IEF-Western blot assay, 0.73 (0.58-0.87) for msAFP by GISA, 0.89 (0.80-0.97) for msAFP percentage by IEF-Western blot assay, and 0.74 (0.59-0.89) for msAFP percentage by GISA. CONCLUSIONS: Both the serum concentration and percentage of msAFP are potential diagnostic markers for HCC with nondiagnostic AFP. GISA can quantify a specific glycoform of a serologic marker. PMID- 12089171 TI - Performance of a revised cardiac troponin method that minimizes interferences from heterophilic antibodies. AB - BACKGROUND: Recent guidelines for use of cardiac troponin to detect cardiac damage and for cardiovascular risk stratification have made increasingly sensitive troponin assays important. Troponin assays continue to be plagued by interferences caused by heterophilic antibodies (HAs). We evaluated the performance of a revised cardiac troponin I (cTnI) assay designed to have increased analytical sensitivity and to minimize the effect of HAs. METHODS: The revised Dade Behring Dimension cTnI assay was evaluated according to NCCLS EP5-A at five institutions. Plasma samples from 14 309 patients were assayed by the original Dimension cTnI assay. To identify samples that may have interfering HAs, samples with values >1.4 microg/L were reanalyzed on the Dade Behring Stratus CS cTnI assay. Samples with possible interfering antibodies were also analyzed before and after selective absorbance studies on the revised Dade Behring Dimension cTnI assay. RESULTS: The limit of quantification in the revised method was 0.1 microg/L with imprecision (CV) of 11-17% at 0.1 microg/L. Values correlated well with the Stratus CS cTnI method: revised = 1.06(original) + 0.01; r = 0.98, S(y/x) = 0.25 microg/L). Falsely increased results consistent with myocardial infarction by the original Dimension cTnI assay and presumably attributable to HAs were identified in 0.17% of all patients with samples submitted for cTnI analysis. The revised Dimension cTnI assay eliminated the interference in 17 of 25 samples identified and greatly decreased the interference in the other 8. CONCLUSIONS: The revised Dimension cTnI method greatly minimizes the effect of interfering HAs. It also exhibits analytical performance characteristics consistent with recent guidelines for use of this assay to detect cardiac damage. PMID- 12089172 TI - Quantification of pro-B-type natriuretic peptide and its products in human plasma by use of an analysis independent of precursor processing. AB - BACKGROUND: Measurement of cardiac natriuretic peptides or their precursors in plasma appears promising in the diagnosis of heart failure. However, the currently available assays to measure pro-B-type natriuretic peptide (proBNP) derived peptides have produced grossly discrepant results. METHODS: We treated plasma with trypsin before assay and used in the assay an antibody specific for a processing-independent epitope of human proBNP. We then determined the total concentration of proBNP and its products in healthy volunteers and heart-failure patients. RESULTS: The antiserum produced (no. 98192) required an intact proBNP NH2 terminus for binding and displayed a high titer, index of heterogeneity, and binding affinity, implying that the RIA was monospecific and highly sensitive. Preanalytical tryptic treatment of plasma cleaved proBNP forms to release the N terminal 1-21 fragment. Furthermore, enzymatic treatment of plasma also was efficient in avoiding nonspecific interference from plasma proteins, making it an expedient alternative to extraction. In healthy individuals, the total proBNP concentrations increased with age from 2.0 pmol/L (range, 0-15 pmol/L; ages 51-65 years) to 22 pmol/L (range, 3-40 pmol/L; ages 66-88 years; P <0.0001). The increase in plasma proBNP in the elderly, however, also seems to reflect the prevalence of cardiac disease. Plasma concentrations in patients with heart failure were all markedly increased [median, 89 pmol/L (range, 29-659) vs 1.0 pmol/L (range, 0-16) in age-matched controls; P <0.0001]. CONCLUSIONS: The processing-independent analysis measures the total proBNP product irrespective of the degree of proBNP processing. The results show that proBNP and its products circulate in low picomolar concentrations in healthy individuals. PMID- 12089173 TI - Relationship between genetic polymorphisms of alcohol-metabolizing enzymes and changes in risk factors for coronary heart disease associated with alcohol consumption. AB - BACKGROUND: There are large individual variations in the responses of risk factors for coronary heart disease to alcohol consumption. To clarify the factors responsible for these individual variations, we studied the relationship between blood pressure, serum lipids, and uric acid and the genetic polymorphisms of alcohol dehydrogenase (ADH) 2 and aldehyde dehydrogenase (ALDH) 2 in alcohol drinkers. METHODS: We examined 133 male workers who drank >300 g of alcohol per week. Information regarding lifestyle habits was obtained by questionnaire. The ADH2 genotype was determined by PCR and subsequent digestion with MaeIII. The ALDH2 genotype was determined based on amplified product length polymorphisms. RESULTS: When the workers were divided into three groups: the ADH2(1)/2(1), ADH2(1)/2(2), and ADH2(2)/2(2) groups, the mean triglycerides and gamma-glutamyl transpeptidase concentrations were significantly higher in the ADH2(2)/2(2) group than in the ADH2(1)/2(1) group. In addition, multiple logistic regression analysis showed that the frequencies of individuals whose systolic blood pressure, triglycerides, and uric acid values were in the highest one third were significantly higher in the ADH2(2)/2(2) group than in the ADH2(1)/2(1) group. In contrast, no difference was observed between the ALDH2(1)/2(1) and (ALDH2(1)/2(2) + ALDH2(2)/2(2)) groups with regard to the mean value of any variable and to the frequency of individuals with any variable value in the highest one third. CONCLUSION: Individuals with the ADH2(1)/2(1) genotype might suffer fewer negative effects of drinking. PMID- 12089174 TI - Determination of sulpiride by capillary electrophoresis with end-column electrogenerated chemiluminescence detection. AB - BACKGROUND: Capillary electrophoresis (CE) with tris(2,2'-bipyridyl)ruthenium(II) [Ru(bpy)3(2+)]-electrogenerated chemiluminescence (ECL) detection is a promising method for clinical analysis. In this study, a method combining CE with Ru(bpy)3(2+) ECL (CE-ECL) detection that can be applied to amine-containing clinical species was developed, and the performance of CE-ECL as a quantitative method for determination of sulpiride in human plasma or urine was evaluated. METHODS: Sulpiride was separated by capillary zone electrophoresis in uncoated fused-silica capillaries [50 cm x 25 microm (i.d.)] filled with phosphate buffer (pH 8.0) and a driving voltage of +15 kV, with end-column Ru(bpy)3(2+) ECL detection. A platinum disc electrode was used as working electrode. Sulpiride in human plasma or urine samples (100 microL) was extracted by a double-step liquid liquid extraction procedure, dried under nitrogen at 35 degrees C in a water bath, and reconstituted with 100 microL of filtered water. The extraction solvent was ethyl acetate-dichloromethane (5:1 by volume). RESULTS: Under optimum conditions (pH 8.0 phosphate buffer, injection for 6 s at 10 kV, and +1.2 V as detection potential), separation of sulpiride was accomplished within 4 min. The calibration curve was linear over a concentration range of 0.05-25.0 micromol/L, and the limit of detection was 2.9 x 10(-8) mol/L for sulpiride. Intra- and interday CVs for ECL intensities were <6%. Extraction recoveries of sulpiride were 95.6-101% with CVs of 2.9-6.0%. The method was clinically validated for patient plasma and urine samples. CONCLUSIONS: CE combined with Ru(bpy)3(2+) ECL is reproducible, precise, selective, and enables the analysis of sulpiride in human plasma and urine. It thus is of value for rapid and efficient analysis of amine-containing analytes of clinical interest. PMID- 12089175 TI - Evaluation of a no-pretreatment cyclosporin A assay on the Dade Behring dimension RxL clinical chemistry analyzer. AB - BACKGROUND: Monitoring whole-blood concentrations of cyclosporin A (CsA) is common practice in the management of solid organ and bone marrow transplant recipients. In a multicenter study we evaluated a new, direct (no pretreatment) CsA assay on the Dade Behring Dimension RxL system and compared results with those from the Abbott TDx CsA immunoassay and a HPLC method. METHODS: Whole-blood samples from heart (n = 111; 35 patients), liver (n = 201; 44 patients), kidney (n = 279; 65 patients), and miscellaneous organ (n = 77; 12 lung, 12 bone marrow, 5 kidney/pancreas, and 1 pancreas patient) recipients were obtained from patient populations of the participating institutions. Routine clinical monitoring of CsA was performed using either the TDx method or HPLC. RESULTS: The minimum detectable concentration of CsA averaged 9.4 microg/L, and the lower limit of quantification was 30 microg/L. The method was linear from 30 to 500 microg/L. Cross-reactivity with seven different CsA metabolites ranged from 0.0% to 5.7% for the Dimension RxL assay compared with 0.4-15.9% for the TDx assay. Total imprecision (CV) averaged 6.2%, and within-run imprecision averaged 4.9%. Passing Bablok linear regression analyses of all samples from two sites yielded the following: RxL = 0.81 x TDx - 16.8; and RxL = 1.12 x HPLC - 1.7. CONCLUSIONS: The Dade Behring CsA assay for the random-access Dimension platform offers adequate performance characteristics for routine clinical use, does not require a manual pretreatment step, and demonstrates less cross-reactivity with CsA metabolites than another commonly used immunoassay. PMID- 12089176 TI - Biochemical markers and hematologic indices in the diagnosis of functional iron deficiency. AB - BACKGROUND: The hypochromic red cell is a direct indicator of functional iron deficiency (ID) in contrast to the majority of biochemical markers, which measure functional ID indirectly via iron-deficient erythropoiesis. The aim of this study was to evaluate the extent to which these biochemical markers can distinguish ID from anemia of chronic disease (ACD) as well as from the combined state of functional ID/ACD, using red cell hemoglobinization as the gold standard. METHODS: We studied 442 patients with various disease-specific anemias and 154 nonanemic patients. As indicators of red cell hemoglobinization, we measured the reticulocyte hemoglobin content (CHr) and the proportion of hypochromic red cells (HYPO), using an Advia 120 hematology analyzer. Ferritin, transferrin, transferrin saturation, and the concentration of the soluble transferrin receptor (sTfR) were determined by ELISA and immunoturbidimetric assay. The sTfR/log ferritin ratio (sTfR-F index) was used as an additional marker for biochemical identification of iron-deficient erythropoiesis. RESULTS: In a control group (n = 71), the 2.5 percentile values were 28 pg for CHr and 5% for HYPO. These values were used to indicate unimpaired red cell hemoglobinization and absence of functional ID. In patients with deficient red cell hemoglobinization but no acute phase response (APR), iron-deficient erythropoiesis was indicated by serum ferritin and sTfR-F index values < or =20.8 microg/L and >1.5, respectively. Corresponding values in patients with APR were < or =61.7 microg/L and >0.8, respectively. The positive likelihood ratios for the biochemical markers and the sTfR-F index for identifying iron-restricted erythropoiesis in patients with and without APR were 2.6-6.9 and 4.3-16.5, respectively. CONCLUSION: In APR patients, biochemical markers demonstrate weaknesses in the diagnosis of functional ID as defined by hematologic indices. Use of diagnostic plots to illustrate the relationship between the sTfR-F index and CHr allows the progression of ID to be identified, regardless of whether an APR is present. PMID- 12089177 TI - Technical evaluation of a new immunoradiometric and a new immunoluminometric assay for thyroglobulin. AB - BACKGROUND: After removal of differentiated thyroid carcinoma (DTC), serum thyroglobulin (Tg) can indicate persistent or recurrent disease. We describe two novel two-step assays designed to measure low Tg concentrations. METHODS: We evaluated prototypes of the new IRMA, DYNOtest Tg-pluS, and the new immunoluminometric assay (ILMA), LUMItest) Tg-pluS. In the first step, a high salt incubation buffer leads to dissociation of Tg-Tg antibody complexes in serum and is intended to reduce nonspecific interference and interference of potential Tg autoantibodies in the system. We studied recovery of human Tg (from thyroid glands) added to horse serum. We also studied 58 patients with DTC in whom Tg values under thyroid-stimulating hormone (TSH) suppression and TSH stimulation (without thyroxine) were available. RESULTS: The detection limits were 0.04 microg/L Tg for the IRMA and 0.02 microg/L for the ILMA. Intraassay imprecision (CV) was <10% over the range of the calibration curve in both assays. The day-to day CV was <20% at 0.2 microg/L for the IRMA and at 0.06 microg/L for the ILMA. No high-dose hook effect was seen with up to 200 000 microg/L added Tg or in dilutions of 12 patient sera with Tg values of 307-38 880 microg/L. Mean recovery of 50 microg Tg/L was 96% in those patients. Among 77 samples with Tg antibody values of 65.2-8150 kilounits/L, recovery by the IRMA was disturbed in 7 cases (9%) and by the ILMA in 9 cases (12%). Tg increased as measured in both assays in 50 of 58 patients after thyroxine withdrawal. CONCLUSIONS: The new assays have improved precision for Tg <1 microg/L, and even low measured Tg concentrations respond physiologically to thyroxine withdrawal. The assays are free of a high dose hook effect up to a Tg concentration of at least 38 000 microg/L and may further reduce Tg antibody interference. PMID- 12089178 TI - Computer-supported interpretation of protein profiles after capillary electrophoresis. AB - BACKGROUND: Electrophoretic patterns of proteins in serum/plasma are useful in the diagnosis and evaluation of many diseases. Capillary zone electrophoresis (CZE) allows rapid and automated protein separation and produces digital absorbance data, appropriate for mathematical analysis. We previously demonstrated success in detection of monoclonal immunoglobulins in such a system. This study tests new algorithms to produce rapid standardized computer-supported interpretation of the entire electropherogram. METHODS: Data from Beckman Paragon CZE 2000 electropherograms were compared with quantitative protein data from >800 routine clinical samples. Algorithms were designed to produce semiquantitative analyses of major proteins and to define different patterns of inflammation based on the electropherogram. RESULTS: The algorithms produced reliable semiquantitative evaluations of prealbumin, albumin, alpha1-antitrypsin, haptoglobin, and transferrin, but were less accurate for alpha1-acid glycoprotein. Some genetic variants of albumin and deficiency variants of alpha1 antitrypsin were easily recognized. Complex clinical traits such as degree and type of inflammation could be evaluated. When used together with previously developed algorithms addressing immunoglobulins, the new algorithms provide relevant clinical interpretation. Selected outputs indicate the need for reflex testing or evaluation by specialists. CONCLUSIONS: Automation of both electrophoresis and interpretation can provide a rapid, inexpensive, standardized analysis that can hopefully improve the diagnostic information and clinical outcome for large groups of patients. It also provides objective criteria for clinical interpretations, to be validated or adjusted in future clinical studies. PMID- 12089179 TI - Serum folate by two methods in pregnant women carrying fetuses with neural tube defects. PMID- 12089180 TI - Direct spectrophotometric method for measurement of bilirubin in newborns: comparison with HPLC and an automated diazo method. PMID- 12089181 TI - Release characteristics of cardiac biomarkers and ischemia-modified albumin as measured by the albumin cobalt-binding test after a marathon race. PMID- 12089182 TI - Perinatal reference intervals for plasma homocysteine and factors influencing its concentration. PMID- 12089183 TI - C-reactive protein kinetics in newborns: application of a high-sensitivity analytic method in its determination. PMID- 12089184 TI - Rhabdomyolysis-related renal tubular damage studied by proton nuclear magnetic resonance spectroscopy of urine. PMID- 12089185 TI - Analytical characteristics of the AxSYM cardiac troponin I and creatine kinase MB assays. PMID- 12089186 TI - Differential detection of skeletal troponin I isoforms in serum of a patient with rhabdomyolysis: markers of muscle injury? PMID- 12089187 TI - Preanalytical influences on the measurement of ghrelin. PMID- 12089188 TI - Biological variation of glycohemoglobin. PMID- 12089189 TI - Soluble transferrin receptor (sTfR), ferritin, and sTfR/log ferritin index in anemic patients with nonhematologic malignancy and chronic inflammation. PMID- 12089190 TI - Development and evaluation of a PCR-based, line probe assay for the detection of 58 alleles in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. PMID- 12089191 TI - Effect of corticosteroid therapy on serum cystatin C and beta2-microglobulin concentrations. PMID- 12089192 TI - Serum acylcarnitines and vitamin B12 deficiency. PMID- 12089193 TI - Dried-serum spot assay for folate. PMID- 12089194 TI - Light chain proteinuria and lysozymuria in a patient with acute monocytic leukemia. PMID- 12089195 TI - Practical method for estimating the frequency of specimen mix-up in clinical chemistry laboratories. PMID- 12089197 TI - Guidelines and recommendations in laboratory medicine. PMID- 12089196 TI - IgG anti-transglutaminase autoantibodies in systemic lupus erythematosus and Sjogren syndrome. PMID- 12089198 TI - Alpha1-microglobulin is stable in human urine ex vivo. PMID- 12089199 TI - Serum cystatin C as a marker of kidney dysfunction in an elderly population. PMID- 12089203 TI - Visual navigation in desert ants Cataglyphis fortis: are snapshots coupled to a celestial system of reference? AB - Central-place foraging insects such as desert ants of the genus Cataglyphis use both path integration and landmarks to navigate during foraging excursions. The use of landmark information and a celestial system of reference for nest location was investigated by training desert ants returning from an artificial feeder to find the nest at one of four alternative positions located asymmetrically inside a four-cylinder landmark array. The cylindrical landmarks were all of the same size and arranged in a square, with the nest located in the southeast corner. When released from the compass direction experienced during training (southeast), the ants searched most intensely at the fictive nest position. When instead released from any of the three alternative directions of approach (southwest, northwest or northeast), the same individuals instead searched at two of the four alternative positions by initiating their search at the position closest to the direction of approach when entering the landmark square and then returning to the position at which snapshot, current landmark image and celestial reference information were in register. The results show that, in the ants' visual snapshot memory, a memorized landmark scene can temporarily be decoupled from a memorized celestial system of reference. PMID- 12089204 TI - Accumulation of the mitochondrial form of the sulphydryl oxidase Erv1p/Alrp during the early stages of spermatogenesis. AB - In this study, we investigated the expression of the mammalian FAD-dependent sulphydryl oxidase Erv1p/Alrp in the rat and mouse and during mouse spermatogenesis. Up to three forms of Alrp were identified in protein extracts from different tissues and organs, but very little enzyme was present in blood samples. The three forms of Alrp represent the full-length protein of 23 kDa and fragments of 21 kDa and 15 kDa. All forms of Alrp were assembled into dimers in vivo. In contrast to samples from other organs, the protein analysis of mouse testis identified predominantly full-length 23 kDa Alrp. This finding prompted us to investigate in more detail the expression of Alrp during spermatogenesis. Testis samples of individual mice from postnatal days 13-29 were probed with an antibody specific for mammalian Alrp. In addition, cells from whole testis preparations were fractionated on a bovine serum albumin column gradient. Protein expression of mouse Alrp was compared with those of testis-specific cyritestin, the cytoskeleton marker actin and mitochondrial subunit Vb of cytochrome oxidase and cytochrome c. These studies demonstrated a specific accumulation of full length mouse Alrp during the early stages of spermatogenesis. The highest levels of Alrp were found in spermatogonia and primary spermatocytes. Levels of expression of Alrp did not correlate with the synthesis of components of the respiratory chain, indicating that full-length Alrp in the mitochondria of spermatogonia and spermatocytes has another function in addition to its role in oxidative phosphorylation. PMID- 12089205 TI - Functional morphology of proximal hindlimb muscles in the frog Rana pipiens. AB - Musculoskeletal models have become important tools in understanding motor control issues ranging from how muscles power movement to how sensory feedback supports movements. In the present study, we developed the initial musculotendon subsystem of a realistic model of the frog Rana pipiens. We measured the anatomical properties of 13 proximal muscles in the frog hindlimb and incorporated these measurements into a set of musculotendon actuators. We examined whether the interaction between this musculotendon subsystem and a previously developed skeleton/joint subsystem captured the passive behavior of the real frog's musculoskeletal system. To do this, we compared the moment arms of musculotendon complexes measured experimentally with moment arms predicted by the model. We also compared sarcomere lengths measured experimentally at the starting and take off positions of a jump with sarcomere lengths predicted by the model at these same limb positions. On the basis of the good fit of the experimental data, we used the model to describe the multi-joint mechanical effects produced by contraction of each hindlimb muscle and to predict muscle trajectories during a range of limb behaviors (wiping, defensive kicking, swimming and jumping). Through these analyses, we show that all hindlimb muscles have multiple functions with respect to accelerating the limb in its three-dimensional workspace and that the balance of functions depends greatly on limb configuration. In addition, we show that muscles have multiple, task-specific functions with respect to the type of contraction performed. The results of this study provide important data regarding the multifunctional role of hindlimb muscles in the frog and form a foundation upon which additional model subsystems (e.g. neural) and more sophisticated muscle models can be appended. PMID- 12089206 TI - S- and C-start escape responses of the muskellunge (Esox masquinongy) require alternative neuromotor mechanisms. AB - The startle response is a model system for examining the neural basis of behavior because of its relatively simple neural circuit organization and kinematic pattern. In fishes, the two primary types of startle behavior differ in their initial movements. In the C-start type of startle, the fish bends into a C shape, while the S-start involves an S-shaped body bend. Although considerable research has focused on determining how the C-start is generated neurally, S-start neurobiology has not been examined. I quantify the kinematics and electromyographic patterns of the initial movements of the C-start and S-start behaviors of the muskellunge (Esox masquinongy) to test three hypotheses for how the S-start is generated. (i) The S-start is generated by the same motor neural circuit as the C-start, but passive bending of the tail causes the body to take on an S shape. (ii) The S-start is generated by the same motor neural circuit as undulatory swimming. (iii) The S-start is generated by an independent neural mechanism from that used either in the C-start or in undulatory swimming. Results from kinematics and muscle activity patterns support the third hypothesis. In the muskellunge, the S-start is a high-performance startle behavior with peak angular velocity and peak angular acceleration of its initial bending comparable with those of the C-start and higher than would be expected for undulatory swimming. The S-start motor pattern, however, is distinct from the C-start motor pattern in having simultaneous muscle activity anteriorly on one side of the body and posteriorly on the opposite side. In contrast, the C-start is characterized by simultaneous unilateral muscle activity along the full length of the body. Alternative models are proposed for S-start neural circuit organization involving reticulospinal and local control of muscle activity. PMID- 12089207 TI - Spectral reflectance and directional properties of structural coloration in bird plumage. AB - Bird plumage is coloured both by pigments and by spectrally selective interference in the light reflected from feather barbs. These barbs are composites of high- and low-refractive-index materials, and light is reflected at refractive index boundaries. The spatial structure determines the wavelengths where constructive interference occurs and, hence, the spectral tuning. The spectral tuning of interference colours often varies with angles of illumination and reflection, which produces iridescence. Iridescence and other optical effects mean that interference coloration looks different from pigmentation and is visually striking. To study the optical and visual properties of structural plumage colours, we recorded the reflectance spectra of feathers and in particular their directional properties. A fixed spot on a feather was viewed from a fixed position, whilst the feather orientation and the angle of illumination were varied. We recognise two main types of coloration, 'directional' and 'diffuse'. Within these types, there is considerable variation, and five examples illustrate some features of structural plumage colours and suggest how their optical and visual properties can be measured and described. PMID- 12089208 TI - Radula-centric and odontophore-centric kinematic models of swallowing in Aplysia californica. AB - Two kinematic models of the radula/odontophore of the marine mollusc Aplysia californica were created to characterize the movement of structures inside the buccal mass during the feeding cycle in vivo. Both models produce a continuous range of three-dimensional shape changes in the radula/odontophore, but they are fundamentally different in construction. The radulacentric model treats the radular halves as rigid bodies that can pitch, yaw and roll relative to a fixed radular stalk, thus creating a three-dimensional shape. The odontophore-centric model creates a globally convex solid representation of the radula/odontophore directly, which then constrains the positions and shapes of internal structures. Both radula/odontophore models are placed into a pre-existing kinematic model of the I1/I3 and I2 muscles to generate three-dimensional representations of the entire buccal mass. High-temporal-resolution, mid-sagittal magnetic resonance (MR) images of swallowing adults in vivo are used to provide non-invasive, artifact-free shape and position parameter inputs for the models. These images allow structures inside the buccal mass to be visualized directly, including the radula, radular stalk and lumen of the I1/I3 cavity. Both radula-centric and odontophore-centric models were able to reproduce two-dimensional, mid-sagittal radula/odontophore and buccal mass kinematics, but the odontophore-centric model's predictions of I1/I3, I2 and I7 muscle dimensions more accurately matched data from MR-imaged adults and transilluminated juveniles. PMID- 12089209 TI - Glutamine synthetase expression in liver, muscle, stomach and intestine of Bostrichthys sinensis in response to exposure to a high exogenous ammonia concentration. AB - A previous study provided evidence that the adaptive strategy used by the teleost fish Bostrichthys sinensis (sleeper) for detoxifying ammonia during extended periods of air exposure was to synthesize and store glutamine, primarily in the muscle, accompanied by an increase in glutamine synthetase (GSase) activity in liver. The aim of the present study was to assess the effect on GSase expression in various tissues of exposure of B. sinensis to exogenous ammonia. Exogenous ammonia increases internal ammonia concentrations in fish, mimicking environmental situations such as air exposure that preclude loss of ammonia across the gills, and thus triggering alternative mechanisms for ammonia detoxification. The results reveal relatively high levels of GSase activity, not only in liver but also, unexpectedly, in muscle, and even higher levels in intestine and, in particular, stomach. Exposure to ammonia results in significant increases in GSase activity, GSase protein and GSase mRNA levels in all of these tissues except stomach. The amino acid sequences of GSases from liver and stomach deduced from the cDNA sequences are essentially identical and are >97 % identical to the amino acid sequences of GSases from Gulf toadfish (Opsanus beta) and marble goby (Oxyeleotris marmoratus). PMID- 12089210 TI - Substrate utilization during graded aerobic exercise in rainbow trout. AB - A biochemical approach was employed to examine the oxidative utilization of carbohydrate and lipid in red muscle of rainbow trout (Oncorhynchus mykiss) during sustained swimming at 30 and 60% of their critical swimming speed (U(crit); for 2, 15 and 240 min) and during non-sustainable swimming at 90% U(crit) (for 2, 15 and 45 min). Measurements included pyruvate dehydrogenase (PDH) activity, creatine phosphate, ATP, glycogen, glycolytic intermediates, acetyl-CoA, acetyl-, total-, free-, short-chain fatty acyl- and long-chain fatty acyl- carnitine, intramuscular triacylglycerol and malonyl-CoA concentrations, and whole body oxygen consumption ((O)(2)). During the first 2 min at 30 and 60% U(crit), oxidation of endogenous glycogen by PDH activation increased 4- and 8 fold, respectively, yielding 1.5- to 2.5-fold increases in acetyl-CoA and 2- to 6 fold increases in acetyl-carnitine concentrations. Within 15 min, PDH activity returned to control values (153.9+/-30.1 nmol g(-1) wet tissue min(-1)); after 240 min there were small 1.7- to 2.6-fold increases in long-chain fatty acyl carnitine and approx. 50% decreases in malonyl-CoA concentrations, indicating an overall enhancement of lipid oxidation. Sustainable swimming at 30 and 60% U(crit) was further characterized by 1.5- and 2.2-fold increases in M(O(2)), respectively. Non-sustainable swimming at 90% U(crit) was characterized by a sustained tenfold (approx.) elevation of red muscle PDH activity (approx. 1600 nmol g(-1) wet tissue min(-1)). Significant 67% decreases in white muscle creatine phosphate and 73% decreases in glycogen levels, without matching increases in lactate levels, point to significant recruitment of white muscle during high-speed swimming for power production, and the potential export of white muscle lactate to red muscle for oxidation. Overall, sustainable exercise at 30 and 60% U(crit) is supported by approximately equal contributions of carbohydrate (approx. 45%) and lipid (approx. 35%) oxidation, whereas non sustainable swimming is supported primarily by carbohydrate oxidation with only moderate contributions from lipid oxidation. PMID- 12089211 TI - Long-term fasting and realimentation in hypogean and epigean isopods: a proposed adaptive strategy for groundwater organisms. AB - The effects of long-term fasting and subsequent refeeding on digestive physiology and energy metabolism were investigated in a subterranean aquatic crustacean, Stenasellus virei, and in a morphologically similar surface-dwelling species, Asellus aquaticus. Metabolic response to food deprivation was monophasic in A. aquaticus, with an immediate, large decrease in all energy reserves. In contrast, S. virei displayed three successive periods of phosphageno-glucidic, lipidic and, finally, proteo-lipidic-dominant catabolism over the course of the nutritional stress. To represent the responses of subterranean crustaceans to food stress and renutrition, a sequential energy strategy was hypothesized, suggesting that four successive phases (called stress, transition, adaptation and recovery) can be distinguished. Based on these results, a general adaptive strategy for groundwater organisms was proposed. Their remarkable resistance to long-term fasting may be partly explained by (1) a depressed metabolism, during which they mainly subsist on lipid stores, (2) a prolonged state of glycogen- and protein sparing, (3) low energetic requirements and (4) large body stores. In addition, these groundwater species displayed high recovery abilities during refeeding, showing an optimal utilization of available food and a rapid restoration of their body reserves. These adaptive responses might be considered for numerous subterranean organisms as an efficient energy-saving strategy in a harsh and unpredictable environment where fasting (and/or hypoxic) periods of variable duration alternate with sporadic feeding events (and/or normoxic periods). Therefore, food-limited and/or hypoxia-tolerant groundwater species appear to be good examples of animals representing a low-energy system. PMID- 12089213 TI - Effect of environmental temperature on body temperature and metabolic heat production in a heterothermic rodent, Spermophilus tereticaudus. AB - This study quantifies the thermoregulatory ability and energetics of a mammal, the round-tailed ground squirrel Spermophilus tereticaudus, that can relax thermoregulatory limits without becoming inactive. We measured body temperature and metabolic rate in animals exposed for short periods (1 h) to air temperatures ranging from 10 to 45 degrees C and for long periods (8 h) to air temperatures ranging from 10 to 30 degrees C. Within 45 min of exposure to air temperatures ranging from 10 to 45 degrees C, the mean body temperatures of alert and responsive animals ranged from 32.1 degrees C (T(air)=10 degrees C) to 40.4 degrees C (T(air)=45 degrees C). This thermolability provided significant energetic savings below the thermoneutral zone, ranging from 0.63 W (18 %) at 10 degrees C to 0.43 W (43 %) at 30 degrees C. When exposed for 8 h to air temperatures between 10 and 30 degrees C, animals varied their body temperature significantly over time. At all air temperatures, the lowest body temperature (maintained for at least 1 h) was 31.2 degrees C. The highest body temperatures (maintained for at least 1 h) were 33.6 degrees C at 10 degrees C, 35.3 degrees C at 20 degrees C and 36.3 degrees C at 30 degrees C. The energetic savings realized by maintaining the minimum rather than the maximum body temperature was 0.80 W (25 %) at 10 degrees C, 0.71 W (33 %) at 20 degrees C and 0.40 W (47 %) at 30 degrees C. This study demonstrates in several ways the ability of this species to adjust energy expenditure through heterothermy. PMID- 12089212 TI - In the polymorphic ciliate Tetrahymena vorax, the non-selective phagocytosis seen in microstomes changes to a highly selective process in macrostomes. AB - Ciliates use phagocytosis to acquire edible particles. The polymorphic ciliate Tetrahymena vorax appears in two forms ('microstomes' and 'macrostomes'). Transformation of microstomes into macrostomes takes place in the presence of the ciliate Tetrahymena thermophila and enables the macrostome to phagocytose the latter species. The non-specific, constitutive phagocytosis in microstomes thereby changes into a specific inducible process in macrostomes. The purpose of this study was to determine whether the phagocytotic process in macrostomes is specifically aimed at catching T. thermophila. The two forms of phagocytosis represent an interesting model system for studying the mechanism whereby phagosomes are formed. The macrostomal form capture deciliated and ciliated Tetrahymena thermophila, latex beads with diameters of 20.3 and 30.0 microm and small microstomal cells. However, the macrostomes select T. thermophila as a prey when they have the opportunity to choose between deciliated T. thermophila and latex beads and between T. thermophila and microstomes. The non-selective formation of phagosomes seen in microstomes changes to a highly selective process during the transformation to macrostomes. Unlike microstomes, macrostomes do not form a closed vacuole after capturing a latex bead, indicating that mechanical stimulation by the prey does not in itself trigger phagocytosis in the macrostomal form of T. vorax. Although macrostomes captured T. thermophila in preference to microstomes, phagocytosis of microstomes started immediately following capture, indicating that the substance/molecule that triggers the formation of the phagosome is not specific for T. thermophila cells. After capturing a T. thermophila cell, the macrostomal cell, which normally swims in a forward direction, reverses direction and swims backwards for a short time before starting to rotate. Macrostomal cells did not change their swimming pattern after capturing a latex bead. We believe, therefore, that backward swimming is more likely to be related to signals resulting from phagocytosis than from mechanical stimulation of the pouch. Cytochalasin B (10 microg ml(-1)) inhibits phagocytosis in both microstomes and macrostomes, indicating that actin filaments play an active role in phagocytosis in both cell types. The antitubulin drug nocodazole (0.3-30 micromol l(-1)) inhibits the formation of more than one phagosome in the macrostome, indicating that membrane transport to the oral apparatus in macrostomes is guided by microtubules. Nocodazole has no effect on the process of phagocytosis in microstomes. PMID- 12089214 TI - Effect of increased calcium concentration in sea water on calcification and photosynthesis in the scleractinian coral Galaxea fascicularis. AB - The relationship between calcification and photosynthesis in coral was investigated using standard sea water with enhanced calcium concentration. In standard sea water at 23 degrees C with the calcium concentration increased by 2.5 mmol l(-1), incorporation of calcium into the skeleton increased by 30-61 %, depending on the method of data normalisation, and photosynthesis, measured as (14)C incorporation into the tissues, also increased by 87 %. At 29 degrees C, calcium incorporation into the skeleton increased by 54-84 % and (14)C incorporation increased by 32 % when sea water calcium concentration was increased by 5 mmol l(-1). However, photosynthesis measured as net photosynthetic oxygen production did not increase. Similarly there was no change in respiration rate when coral polyps were incubated in high-calcium sea water. It is conjectured that an increase in photorespiration may be responsible for the latter observations. Bisphosphonate has been considered to inhibit calcification but not photosynthesis in corals. We show that bisphosphonate may not inhibit formation of amorphous calcium carbonate and that the inhibition of calcification is possibly illusory. The data are consistent with the trans-calcification model, which suggests that calcification is a source of CO(2) for photosynthesis in corals. PMID- 12089215 TI - Extreme resistance to desiccation and microclimate-related differences in cold hardiness of gall wasps (Hymenoptera: Cynipidae) overwintering on roses in southern Canada. AB - Four species of cynipid wasp of the genus Diplolepis that induce galls on roses (Rosa species) in southern Canada and two species of inquiline cynipid associated with these galls were studied for their cold-hardiness and resistance to water loss and for possible links between these adaptations. Mid-winter-acclimated supranivean D. spinosa and Periclistus pirata had lower supercooling points (-38 to -40 degrees C) and higher hemolymph osmolalities (1760-1849 mosmol kg(-1)) than subnivean D. polita, D. gracilis, D. radicum and Periclistus sp. (-31 to -32 degrees C and 977-1464 mosmol kg(-1), respectively). During a simulated transition from summer/fall to mid-winter conditions, the glycerol concentration of D. spinosa more than tripled, reaching a final value of 0.98 moll(-1), while its supercooling point decreased by 13 degrees C from the initial value of -27.4 degrees C; however, glycerol concentration and supercooling point did not change for the subnivean species. The permeability of the cuticle of all species was extremely low (0.33-1.00 microg h(-1) cm(-2) mmHg(-1) at 5 degrees C and 0 % relative humidity; 1 mmHg=0.133 kPa), even compared with that of desert species; however, there was no difference in cuticular permeability between supranivean and subnivean prepupae. Transition temperatures ranged between 32.3 and 34.6 degrees C; below 30 degrees C, temperature had little effect on rates of water loss for all species (Q(10)=1.13-1.87). PMID- 12089216 TI - Kinematics of plaice, Pleuronectes platessa, and cod, Gadus morhua, swimming near the bottom. AB - The kinematics of plaice (Pleuronectes platessa, L=22.1 cm) and cod (Gadus morhua, L=25.0 cm, where L is total fish length) swimming at various speeds at the bottom and lifted to heights, h, of 10, 50 and 100 mm by a thin-wire grid were measured. For cod, tailbeat frequency, amplitude, body and fin span and propulsive wavelength were unaffected by h and varied with speed as described for fusiform pelagic species. In contrast, the kinematics of plaice was affected by h. Body and fin spans and propulsive wavelength were independent of swimming speed and h. Tailbeat amplitude was independent of swimming speed, but averaged 1.5 cm at h=0 and 2.5 cm at h> or = 10 mm. Plaice tailbeat frequency increased with swimming speed for fish at the bottom but was independent of swimming speed at h=10, 50 and 100 mm, averaging 4.6, 6.0 and 5.8 Hz respectively. Total mechanical power, P, produced by propulsive movements calculated from the bulk momentum form of elongated slender-body theory was similar for cod and plaice swimming at the bottom but, at h> or = 10 mm, P for plaice was larger than that for cod. Plaice support their weight in water by swimming at a small tilt angle. The small changes in swimming kinematics with swimming speed are attributed to decreasing induced power costs to support the weight as speed increases. The contribution of the tail to power output increased monotonically with the tail gap/span ratio, z/B, for z/B=0.23 (h=0 mm) to z/B=1.1 (h=50 mm). The smaller tailbeat amplitude of the tail decreased both z/B and the power output for plaice swimming at the bottom. For the maximum body and fin span of plaice, the contribution to power output increased for local z/B values of 0.044 (h-0 mm) to 0.1 (h=10 mm) and declined somewhat at larger values of z/B. The smaller effect of the bottom on power output of the large-span anterior body sections may result from the resorption of much of the upstream wake at the re-entrant downstream tail. PMID- 12089217 TI - Scaling of rotational inertia in murine rodents and two species of lizard. AB - Because the force required to rotate a body about an axis is directly proportional to its rotational inertia about the axis, it is likely that animals with high rotational inertia would be constrained in their turning abilities. Given that rotational inertia scales with mass(1.67) in geometrically similar animals, whereas the ability to apply torque scales with mass(1.00), larger animals would be expected to have more difficulty turning than smaller animals of similar shape. To determine how rotational inertia scales with body mass, we used the fact that the period of a physical pendulum is proportional to its rotational inertia(0.50), and measured rotational inertia in two groups of vertebrates with greatly different body shapes: murine rodents (Mus domesticus and Rattus norvegicus) and lizards (Iguana iguana and Varanus exanthematicus). Rotational inertia did not deviate significantly from isometric scaling in the murine rodents as a group or in the varanid lizards, scaling with mass(1.63) and mass(1.59), respectively. Although rotational inertia did scale with negative allometry in iguanas and rats alone, with mass(1.56) and mass(1.42), respectively, it still increased much more quickly with increasing mass than the predicted ability to apply torque. This suggests either that these animals are not constrained by rotational inertia because of their relatively small size or that larger rodents and lizards are poorer turners than smaller ones. The murine rodents had a 3.0- to 4.9-fold lower rotational inertia than similarly sized lizards of either species. Given that the basal synapsids had body proportions and limb configurations similar to those of modern lizards, we suggest that the loss of the large muscular tail and elongated body form during the evolution of cynodonts and mammals reduced rotational inertia and probably improved turning ability. PMID- 12089218 TI - Quality of life: it's never too late. PMID- 12089219 TI - Boosting bioavailability of topotecan: what do we gain? PMID- 12089220 TI - Quality of life of long-term survivors of non-small-cell lung cancer. AB - PURPOSE: To describe the quality of life (QOL) among survivors of non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: One hundred forty-two 5-year minimum self-reported disease-free survivors of NSCLC completed QOL instruments (QOL Survivor and Medical Outcomes Study 36-Item Short Form [SF-36]) and assessments of emotional distress (Center for Epidemiologic Studies Depression Scale [CES D]), comorbid disease, and tobacco use. Pulmonary function was assessed with a hand-held spirometer. Multivariate regression methods were used on total QOL Survivor scores and physical (PC) and mental (MC) component scores of the SF-36. RESULTS: The majority (71%) of survivors described themselves as hopeful, and 50% viewed the cancer experience as contributing to positive life changes (QOL Survivor). Comorbidity was common (60% >or= one condition); 22% had distressed mood (CES-D >or= 16). Most were former smokers (76%); 13% continued to smoke. Half had moderate/severe pulmonary distress (forced expired volume in 1 second [FEV1] < 70% of predicted). Regression models including the set of variables (age, sex, living alone, education, smoking status, pulmonary function category, distressed mood, time since diagnosis, and comorbidity) accounted for 37%, 48%, and 29% in the QOL-total, MC, and PC scores, respectively. Primary predictors of lower QOL scores were white ethnicity and distressed mood (CES-D >or= 16) (34% of the variance explained). The primary predictor of lower MC scores was distressed mood (R(2) = 0.45). Lower PC scores were associated with older age, living alone, FEV1 less than 70% of predicted, distressed mood, time since diagnosis, and more comorbid diseases (R(2) = 0.28). CONCLUSION: These findings provide the first description of the QOL of long-term survivors of lung cancer. Risk factors for poorer QOL are strongly linked to distressed mood, which is a potential target for intervention. PMID- 12089221 TI - Prognostic impact of micrometastatic tumor cells in the lymph nodes and bone marrow of patients with completely resected stage I non-small-cell lung cancer. AB - PURPOSE: This study was designed to substantiate the prognostic impact of occult micrometastatic tumor cells in the lymph nodes (LNs) and bone marrow (BM) in stage I non-small-cell lung cancer (NSCLC) patients using cytokeratin (CK) as a micrometastatic marker and the relationship between the micrometastases in the LNs and BM. PATIENTS AND METHODS: A total of 2,432 hilar and mediastinal LNs were removed during surgery from 115 patients with completely resected stage I NSCLC. The LNs were analyzed for micrometastasis using immunohistochemistry with the biclonal anti-CK antibody AE1/AE3. BM aspirates from 115 patients were immunocytochemically stained with the monoclonal anti-CK antibody CK2. RESULTS: CK-positive (CK+) cells were detected in 42 (1.7%) of 2,432 LNs, in 32 (27.8%) of 115 patients, and in 32 (27.8%) of 115 BM aspirates. There was no relationship between the frequencies of CK+ cells in the LNs and in the BM. The patients with CK+ cells in the LNs had a poor prognosis by both univariate (P =.008) and multivariate analyses (P =.01), whereas the presence of CK+ cells in the BM did not allow prediction of survival (P =.32). The prognostic impact of LNs micrometastasis was independent even after adjusting for the status of BM micrometastasis. CONCLUSION: The detection of lymph nodal micrometastatic tumor cells provides an accurate assessment of tumor staging and has powerful prognostic implications for completely resected stage I NSCLC patients. PMID- 12089222 TI - Long-term follow-up of patients treated with paclitaxel/carboplatin-based chemotherapy for advanced non-small-cell lung cancer: sequential phase II trials of the Minnie Pearl Cancer Research Network. AB - PURPOSE: To provide long-term follow-up on the survival of patients with advanced non-small-cell lung cancer treated with paclitaxel/carboplatin-based regimens in a multicenter, community-based setting. PATIENTS AND METHODS: Between March 1995 and April 1998, 321 patients with newly diagnosed stage IIIB or IV non-small-cell lung cancer were treated on sequential phase II trials with the following combination regimens: paclitaxel/carboplatin, paclitaxel/carboplatin/gemcitabine, and paclitaxel/carboplatin/vinorelbine. Details of these three regimens and patient populations have been previously reported. Responding and stable patients continued treatment until tumor progression or for a recommended six treatment courses. RESULTS: After a median follow-up of 58 months (minimum follow-up, 40 months), the median survival for the entire group of patients was 8.6 months, with actual 1-, 2-, and 3-year survival rates of 40%, 19%, and 7%, respectively. The actuarial 4-year survival rate for the entire group was 4%. No statistically significant differences in survival were seen among the three regimens. Administration of all three regimens was feasible in a community-based setting; however, myelosuppression and hospitalizations for treatment of neutropenia/fever were more frequent with the three-drug regimens. CONCLUSION: Paclitaxel/carboplatin-based regimens, in addition to prolonging median survival and improving 1-year survival, result in substantial improvements in the 2-year survival of patients with advanced non-small-cell lung cancer when compared retrospectively with supportive care or traditional cisplatin-based regimens. In these sequential phase II trials, we did not demonstrate any advantages of three drug regimens when compared with paclitaxel/carboplatin. Because few patients remain alive after 4 years with any of these chemotherapy regimens, future treatment improvements will require the introduction of novel agents. PMID- 12089223 TI - Increased oral bioavailability of topotecan in combination with the breast cancer resistance protein and P-glycoprotein inhibitor GF120918. AB - PURPOSE: We discovered that breast cancer resistance protein (BCRP), a recently identified adenosine triphosphate-binding cassette drug transporter, substantially limits the oral bioavailability of topotecan in mdr1a/1b(-/-) P glycoprotein (P-gp) knockout and wild-type mice. GF120918 is a potent inhibitor of BCRP and P-gp. The aim was to increase the bioavailability of topotecan by GF120918. PATIENTS AND METHODS: In cohort A, eight patients received 1.0 mg/m(2) oral topotecan with or without coadministration of one single oral dose of 1,000 mg GF120918 (day 1 or day 8). In cohort B, eight other patients received 1.0 mg/m(2) intravenous topotecan with or without 1,000 mg oral GF120918 to study the effect of GF120918 on the systemic clearance of topotecan. RESULTS: After oral topotecan, the mean area under the plasma concentration-time curve (AUC) of total topotecan increased significantly from 32.4 +/- 9.6 microg.h/L without GF120918 to 78.7 +/- 20.6 microg.h/L when GF120918 was coadministered (P =.008). The mean maximum plasma concentration of total topotecan increased from 4.1 +/- 1.5 microg/L without GF120918 to 11.5 +/- 2.4 microg/L with GF120918 (P =.008). The apparent bioavailability in this cohort increased significantly from 40.0% (range, 32% to 47%) to 97.1% (range, 91% to 120%) (P =.008). Interpatient variability of the apparent bioavailability was 17% without and 11% with GF120918. After intravenous administration of topotecan, coadministration of oral GF120918 had a small but statistically significant effect on the AUC and systemic clearance of total topotecan but no statistically significant effect on maximum plasma concentration and terminal half-life of total topotecan. CONCLUSION: Coadministration of the BCRP and P-gp inhibitor GF120918 resulted in a significant increase of the systemic exposure of oral topotecan. The apparent oral bioavailability increased from 40.0% without to 97.1% with GF120918. PMID- 12089224 TI - Phase II study of carboplatin in children with progressive low-grade gliomas. AB - PURPOSE: To assess the rate of tumor response and activity of carboplatin in stabilizing the growth of progressive low-grade gliomas. PATIENTS AND METHODS: Eligible patients received carboplatin 560 mg/m(2) intravenously every 4 weeks for 1 year after maximum tumor response or until disease progression or unacceptable toxicity. RESULTS: Between October 1993 and October 2000, 81 children (median age, 79 months; range, 6 to 204) were enrolled onto this study. Patients received a median of 11 cycles of carboplatin (range, one to 29). Median follow-up from the time of enrollment was 55 months (range, 10 to 93). The overall objective response (complete response [CR] + partial response [PR] + minor response [MR]) and disease stabilization (CR + PR + stable disease + MR) rates to carboplatin treatment were 28% (95% confidence interval [CI], 18% to 38%) and 85% (95% CI, 74% to 93%), respectively. Eleven and 14 patients suffered progressive disease on study and after stopping therapy, respectively. Toxicity was predominantly myelosuppression and included grade 3/4 neutropenia in 56 patients and grade 3/4 thrombocytopenia in 40 patients. The 3-year failure-free survival (FFS) and overall survival (OS) for all patients were 64% (95% CI, 54% to 76%) and 84% (95% CI, 76% to 93%), respectively. Patients with diencephalic tumors had inferior FFS and OS compared with those with tumor at other sites (38% v 74% for FFS, P =.011; 54% v 91% for OS, P =.004). Neurofibromatosis type 1 patients with progressive low-grade glioma had a significantly better OS (95% v 80%; P =.052). CONCLUSION: Carboplatin, in the schedule used in this study, produced disease stabilization or improvement in a majority of children with progressive low-grade glioma, with manageable toxicity. Improved treatment strategies are particularly required for patients with diencephalic tumors. PMID- 12089225 TI - Outcomes of growth hormone replacement therapy in survivors of childhood acute lymphoblastic leukemia. AB - PURPOSE: Little is known about the long-term efficacy or adverse effects of growth hormone (GH) replacement therapy in survivors of childhood acute lymphoblastic leukemia (ALL) who have GH deficiency. We investigated the adult height of patients who had received GH and estimated their risk of leukemia relapse or development of a second malignancy. PATIENTS AND METHODS: Of 910 patients treated for ALL at a single institution, 47 had received GH replacement therapy. The linear growth of these 47 patients was retrospectively evaluated. Their risk of leukemia relapse or second malignancy was compared with that of survivors who did not undergo GH therapy. RESULTS: The median height SD score at the start of GH therapy had decreased by 1.0 since the time of diagnosis of ALL. After a median duration of 4.5 years of GH therapy, adult height SD scores improved and approached height SD scores at the time of diagnosis of ALL. The median adult height for male patients was 173.2 cm (range, 157 to 191.9 cm), and for female patients, it was 158.1 cm (range, 141 to 168 cm). None of the patients developed adverse effects requiring discontinuation of GH treatment. At the 7 year and 11-year landmarks in continuous hematologic remission, there was no statistical evidence that GH therapy was associated with leukemia relapse or development of a second malignancy. CONCLUSION: This study suggests that GH replacement therapy is safe and efficacious for the correction of GH deficiency in survivors of childhood ALL. PMID- 12089226 TI - Cisplatin, gemcitabine, and ifosfamide as weekly therapy: a feasibility and phase II study of salvage treatment for advanced transitional-cell carcinoma. AB - PURPOSE: We investigated the feasibility, safety, and antitumor activity of weekly gemcitabine given in combination with low doses of cisplatin and ifosfamide in previously treated patients with advanced transitional-cell carcinoma (TCC) of the urothelium. PATIENTS AND METHODS: Patients with measurable, metastatic or unresectable TCC who had received one or two prior chemotherapy regimens were eligible. On a 28-day course, doses of cisplatin 30 mg/m(2), gemcitabine 800 mg/m(2), and ifosfamide 1 g/m(2) were given on day 1 and then repeated on day 8 and day 15 unless there was dose-limiting hematologic toxicity. RESULTS: Fifty-one patients were registered; 10 patients participated in a pilot study, after which 41 patients were registered onto the phase II protocol. Forty-eight patients (94.1%) had dose-limiting hematologic toxicity on day 8 or day 15. Nonhematologic toxicity of grade 3 or greater consisted mainly of nausea and vomiting (seven patients, 13.7%) and infection (seven patients, 13.7%). Responses could be assessed in 49 of 51 eligible patients; two complete responses (4.1%) and 18 partial responses (36.7%) were observed for an overall response rate of 40.8% (exact 95% confidence interval, 27% to 56%). CONCLUSION: This regimen of cisplatin, gemcitabine, and ifosfamide is not feasible for weekly administration because of hematologic toxicity. Nevertheless, there was promising activity with only two doses per 28-day cycle. On the basis of these results, we have initiated a phase II trial of this combination given as a single dose every 14 days in patients with untreated, metastatic urothelial carcinoma. PMID- 12089227 TI - Incipient angiogenesis in Barrett's epithelium and lymphangiogenesis in Barrett's adenocarcinoma. AB - PURPOSE: Barrett's esophagus (BE), a precancerous condition for Barrett's adenocarcinoma, is classically characterized by flames of salmon-colored mucosa extending into normal pale esophageal mucosa. This flaming is thought to be a consequence of continuous erosis of mucosa caused by chronic reflux. Another characteristic feature of Barrett's adenocarcinoma patients is the frequent development of lymph node metastases. We addressed whether onset of angiogenesis occurs in BE and if the lymphatic system might provide a route for Barrett's adenocarcinoma cells to infiltrate regular lymph nodes. PATIENTS AND METHODS: Fifteen surgically resected Barrett's dysplasia or adenocarcinoma patients were included. Immunohistochemistry and a modified whole mount analysis were used. RESULTS: The incipient angiogenesis originates from the pre-existing vascular network in the lamina propria and infiltrates Barrett's epithelium, giving its ominous salmon-red color. Barrett's epithelium-specific goblet cells express vascular endothelial growth factor (VEGF)-A. The immature blood vessels show a relative absence of smooth muscle actin (SMA)-positive mural cells and express VEGF receptor (VEGFR)-2 and matrix metalloproteinase (MMP)-9 on their exterior. Coexpression of VEGF-C and its receptor VEGFR-3 on lymphatic vessels is demonstrated. CONCLUSION: BE is strongly neovascularized not eroded. This novel concept of a molecular mechanism of the origin of BE might emphasize why precancerous BE can give rise to the more cancerous dysplasia and Barrett's adenocarcinoma stages. In addition, adenocarcinoma cells induce lymphangiogenesis. The new lymphangiogenic vessels might provide a systemic route for adenocarcinoma cells to invade circulation and induce lymph node metastasis. PMID- 12089228 TI - Alteration of p53 pathway in squamous cell carcinoma of the head and neck: impact on treatment outcome in patients treated with larynx preservation intent. AB - PURPOSE: To identify the role of p53 pathway alteration(s) as predictors of treatment outcome in patients with advanced, resectable, squamous cell carcinoma (SCC) of the larynx and pharynx treated with larynx preservation (LP) intent. PATIENTS AND METHODS: Seventy-one patients treated on two consecutive LP protocols were studied based on availability of representative tissues. We analyzed the expression pattern of p53, its upstream regulator mdm2, and downstream transcriptional target p21/WAF1 by immunohistochemistry. Positive phenotype was defined as >or= 20% of tumor cells showing nuclear immunoreactivity. Results were correlated with treatment outcomes. RESULTS: Positive phenotype was observed in 35 (49%) of 71 cases for p53, in 52 (74%) of 70 for mdm2, and in 37 (54%) of 68 for p21. There was no correlation between p53 phenotype and p21 nuclear accumulation. The mdm2-negative phenotype was most predictive of major response at the primary tumor site (P =.088). p53-positive phenotype was associated with worse local control with LP (LCLP; 49% v 23%, P =.053) and inferior overall survival (OS; 51% v 29%, P =.017) at 5 years. On Cox regression analysis, p53-positive phenotype predicted inferior OS (P =.033) and showed a trend for worse LCLP (P =.102). When analyzed in a multivariate model as continuous variables, p53 showed a stronger correlation with inferior OS (P <.01), and mdm2 was associated with worse OS (P <.01). CONCLUSION: Among the three markers studied, our data support p53 phenotype as the most informative predictor of unfavorable outcomes in the LP setting, and suggest a role for mdm2 phenotype that requires further exploration. Our analysis does not support a p53 dependent mechanism for p21 expression. Prospective and larger studies are necessary before integration of these molecular markers as part of molecular staging and predictors for organ preservation or other outcomes. PMID- 12089229 TI - ChlVPP/EVA hybrid versus the weekly VAPEC-B regimen for previously untreated Hodgkin's disease. AB - PURPOSE: To test the hypothesis that a chemotherapy regimen of relatively low toxicity and 11 weeks' duration (doxorubicin, cyclophosphamide, etoposide, vincristine, bleomycin, and prednisolone [VAPEC-B]) is at least as effective in terms of disease control as 6 months' treatment with chlorambucil, vinblastine, procarbazine, and prednisone/etoposide, vincristine, and doxorubicin (ChlVPP/EVA hybrid), which is associated with a high risk of permanent sterility. PATIENTS AND METHODS: Two hundred eighty-two patients with previously untreated Hodgkin's disease, clinical stages I/II (plus mediastinal bulk and/or B symptoms) and clinical stages III/IV were randomized at three United Kingdom and one Italian center to receive either six monthly cycles of ChlVPP/EVA hybrid or 11 weekly cycles of VAPEC-B. After chemotherapy and a restaging evaluation, radiotherapy was administered to sites of previous bulk or residual radiographic abnormality before patients were observed off treatment. RESULTS: Further accrual to the trial was halted at the planned third interim analysis in September 1996. After a median follow-up of 4.9 years, freedom from progression (FFP), event-free survival (EFS), and overall survival (OS) are all significantly better in the population treated with ChlVPP/EVA than VAPEC-B, where the comparative 5-year results are 82% and 62% (FFP), 78% and 58% (EFS), and 89% and 79% (OS), respectively. The superiority of ChlVPP/EVA was seen in both low-risk and intermediate/high-risk patients, although subset analysis suggested that ChlVPP/EVA and VAPEC-B produce equivalent results in the best-prognosis patients (Hasenclever score or=95% detection limits as determined by probit regression analysis ranged from 1,545 to 2,835 viral genome equivalents/ml of serum (8.6 to 16 RNA copies per assay). The suitability of the assays was exemplified by detection and quantification of viral RNA in serum samples of VHF patients. PMID- 12089243 TI - Clinical evaluation of the ZstatFlu-II test: a chemiluminescent rapid diagnostic test for influenza virus. AB - Exploiting the high sensitivity of the chemiluminescence phenomenon, an accurate and sensitive point-of-care test, called the ZstatFlu-II test (ZymeTx, Inc., Oklahoma City, Okla.), was developed to detect influenza virus infections. The ZstatFlu-II test takes 20 min and requires approximately 2 min of "hands-on" time for operational steps. The ZstatFlu-II test does not distinguish between infections with influenza virus types A and B. ZstatFlu-II test results are printed on Polaroid High-Speed Detector Film, allowing test results to be archived. A prototype version of the ZstatFlu-II test was evaluated during the 2000-to-2001 flu season with 300 nasal aspirate specimens from children at a pediatric hospital. Compared to culture, the ZstatFlu-II test had 88% sensitivity and 92% specificity. The Directigen test had a sensitivity of 75% and a specificity of 93%. The sensitivity of the ZstatFlu-II test was significantly higher than that of the Directigen test (P < 0.0574). PMID- 12089244 TI - Detection and differentiation of Cryptosporidium parasites that are pathogenic for humans by real-time PCR. AB - Cryptosporidiosis is a significant cause of food-borne and waterborne outbreaks of diarrheal diseases. To better understand the route of transmission of Cryptosporidium parasites, a number of genotyping techniques have been developed, based on PCR-restriction fragment length polymorphism or sequencing analysis of antigen, structural, and housekeeping genes. In this study, a real-time assay for the detection of Cryptosporidium oocysts is described. This technique had a detection limit of five oocysts. By melting curve analysis of PCR products with fluorescence-labeled hybridization probes, this technique was able to differentiate five common Cryptosporidium parasites that are pathogenic for humans in a single PCR. We evaluated and validated the test using samples from presently known Cryptosporidium parasites that are pathogenic for humans. This technique provides an alternative molecular tool in epidemiologic studies of human cryptosporidiosis. PMID- 12089246 TI - Characterization of Salmonella serovars by PCR-single-strand conformation polymorphism analysis. AB - PCR-restriction fragment length polymorphism (PCR-RFLP) and PCR-single-strand conformation polymorphism (PCR-SSCP) analyses were carried out on the 1.6-kb groEL gene from 41 strains of 10 different Salmonella serovars. Three HaeIII RFLP profiles were recognized, but no discrimination between the serovars could be achieved by this technique. However, PCR-SSCP analysis of the groEL genes of various Salmonella serovars produced 14 SSCP profiles, indicating the potential of this technique to differentiate different Salmonella serovars (interserovar differentiation). Moreover, PCR-SSCP could differentiate strains within a subset of serovars (intraserovar discrimination), as three SSCP profiles were produced for the 11 Salmonella enterica serovar Enteritidis strains, and two SSCP profiles were generated for the 7 S. enterica serovar Infantis and five S. enterica serovar Newport strains. PCR-SSCP has the potential to complement classical typing methods such as serotyping and phage typing for the typing of Salmonella serovars due to its rapidity, simplicity, and typeability. PMID- 12089245 TI - Rapid and simple approach for identification of Mycobacterium tuberculosis complex isolates by PCR-based genomic deletion analysis. AB - Although the virulences and host ranges differ among members of the Mycobacterium tuberculosis complex (TBC; M. tuberculosis, M. africanum, M. canettii, M. microti, M. bovis, and M. bovis BCG), commercially available molecular assays cannot differentiate these organisms because of the genetic identities of their 16S rRNA gene sequences. Comparative genomic analyses with the complete DNA sequence of M. tuberculosis H37Rv has provided information on regions of difference (RD 1 to RD 16) deleted in members of the TBC other than M. tuberculosis. To determine whether deletion analysis could accurately differentiate members of TBC, we used PCR to assess the presence or absence of specific regions of the genome in 88 well-characterized isolates of M. tuberculosis, M. africanum, M. microti, M. bovis, and M. bovis BCG. The identifications obtained by use of the specific deletion profiles correlated 100% with the original identifications for all TBC members except M. africanum, but further characterization resulted in profiles specific for all members. Although six RD regions were used in the analyses with the original 88 isolates, it was found that the use of RD 1, RD 9, and RD 10 was sufficient for initial screenings, followed by the use of RD 3, RD 5, and RD 11 if the results for any of the first three regions were negative. When 605 sequential clinical isolates were screened, 578 (96%) were identified as M. tuberculosis, 6 (1%) were identified as M. africanum, 8 (1%) were identified as M. bovis, and 13 (2%) were identified as M. bovis BCG. Since PCR-based assays can be implemented in most clinical mycobacteriology laboratories, this approach provides a rapid and simple means for the differentiation of members of TBC, especially M. bovis and M. tuberculosis, when it is important to distinguish between zoonotic sources (i.e., cattle and unpasteurized dairy products) and human sources of tuberculosis disease. PMID- 12089247 TI - Rapid detection of Leishmania infantum infection in dogs: comparative study using an immunochromatographic dipstick test, enzyme-linked immunosorbent assay, and PCR. AB - Current zoonotic visceral leishmaniasis (ZVL) control programs in Brazil include the culling of Leishmania infantum-infected reservoir dogs, a strategy that has failed to prevent a rise of canine and human ZVL cases over the past decade. One of the main reasons this strategy has failed is because of a long delay between sample collection, sample analysis, and control implementation. A rapid, sensitive, and specific diagnostic tool would be highly desirable, because it would allow control interventions to be implemented in situ. We compared an immunochromatographic dipstick test to enzyme-linked immunosorbent assay (ELISA) and PCR for detecting L. infantum infections in dogs from an area of ZVL endemicity in Brazil. The dipstick test was shown to have 61 to 75% specificity and 72 to 77% sensitivity, compared to 100% specificity for both ELISA and PCR and 71 to 88% and 51 to 64% sensitivity for ELISA and PCR, respectively. Of the field samples tested, 92 of 175 (53%), 65 of 175 (37%), and 47 of 175 (27%) were positive by dipstick, ELISA, and PCR, respectively. The positive and negative predictive values for the tested dipstick were 58 to 77% and 75%, respectively. Efforts should be made to develop a more specific dipstick test for diagnosis of leishmaniasis, because they may ultimately prove more cost-effective than currently used diagnostic tests when used in mass-screening surveys. PMID- 12089248 TI - Comparison of an industry-derived LCx Chlamydia pneumoniae PCR research kit to in house assays performed in five laboratories. AB - In a multicenter comparison of PCR assays utilizing 120 quantitated samples of 16 Chlamydia pneumoniae isolates, an LCx research-use-only (RUO) PCR developed by Abbott Laboratories demonstrated 100% sensitivity on 48 samples with >1 copy of DNA per microl of specimen. The sensitivities of five in-house PCR assays ranged from 54 to 94% for the same samples. All six assays showed decreased sensitivities as the DNA copy numbers of the samples decreased. Overall, sensitivities ranged from 68% for the LCx PCR assay to 29% for one of the in house tests. The LCx RUO PCR and three of the five in-house PCR tests reported no false positives with the 24 negative samples. Increasing the number of replicates tested increased the sensitivities of all of the assays, including the LCx PCR. The LCx RUO assay showed high reproducibility for a single technologist and between technologists, with a kappa agreement of 0.77. The within-center agreements of the five in-house PCR tests varied from 0.19 to 0.74 on two challenges of 60 specimens 1 month apart. The LCx C. pneumoniae RUO PCR shows excellent potential for use in clinical studies, which could enable standardization of results in the field. PMID- 12089249 TI - Horizontal transmission of Candida parapsilosis candidemia in a neonatal intensive care unit. AB - This report describes the nosocomial acquisition of Candida parapsilosis candidemia by one of the six premature newborns housed in the same room of a neonatal intensive care unit at the Ospedale Santa Chiara, Pisa, Italy. The infant had progeria, a disorder characterized by retarded physical development and progressive senile degeneration. The infant, who was not found to harbor C. parapsilosis at the time of his admission to the intensive care unit, had exhibited symptomatic conjunctivitis before the onset of a severe bloodstream infection. In order to evaluate the source of infection and the route of transmission, two independent molecular typing methods were used to determine the genetic relatedness among the isolates recovered from the newborn, the inanimate hospital environment, hospital personnel, topically and intravenously administered medicaments, and indwelling catheters. Among the isolates collected, only those recovered from the hands of two nurses attending the newborns and from both the conjunctiva and the blood of the infected infant were genetically indistinguishable. Since C. parapsilosis was never recovered from indwelling catheters or from any of the drugs administered to the newborn, we concluded that (i) horizontal transmission of C. parapsilosis occurred through direct interaction between nurses and the newborn and (ii) the conjunctiva was the site through which C. parapsilosis entered the bloodstream. This finding highlights the possibility that a previous C. parapsilosis colonization and/or infection of other body sites may be a predisposing condition for subsequent C. parapsilosis hematogenous dissemination in severely ill newborns. PMID- 12089250 TI - Characterization of an unusual Mycobacterium: a possible missing link between Mycobacterium marinum and Mycobacterium ulcerans. AB - In an attempt to characterize an unusual mycobacterial isolate from a 44-year-old patient living in France, we applied phenotypic characterizations and various previously described molecular methods for the taxonomic classification of mycobacteria. The results of the investigations were compared to those obtained in a previous study with a set of temporally and geographically diverse Mycobacterium ulcerans (n = 29) and Mycobacterium marinum (n = 29) isolates (K. Chemlal, G. Huys, P.-A. Fonteyne, V. Vincent, A. G. Lopez, L. Rigouts, J. Swings, W. M. Meyers, and F. Portaels, J. Clin. Microbiol. 39:3272-3278, 2001). The isolate, designated ITM 00-1026 (IPP 2000-372), is closely related to M. marinum according to its phenotypic properties, lipid pattern, and partial 16S rRNA sequence. Moreover, fingerprinting by amplified fragment length polymorphism (AFLP) analysis unequivocally classified this strain as a member of the species M. marinum, although it lacked two species-specific AFLP marker bands. However, PCR and restriction fragment length polymorphism analysis based on M. ulcerans specific insertion sequence IS2404 showed the presence of this element in a low copy number in isolate ITM 00-1026. In conclusion, the designation of this isolate as a transitional species further supports the recent claim by Stinear et al. (T. Stinear, G. Jenkin, P. D. Johnson, and J. K. Davies, J. Bacteriol. 182:6322-6330, 2000) that M. ulcerans represents a relatively recent phylogenetic derivative of M. marinum resulting from the systematic acquisition of foreign DNA fragments. PMID- 12089251 TI - Multiplex, quantitative, real-time PCR assay for cytomegalovirus and human DNA. AB - We created a multiplex, quantitative, real-time PCR assay that amplifies cytomegalovirus (CMV) and human DNA in the same reaction tube, allowing for a viral load determination that is normalized to measured human DNA. The assay targets a conserved region of the CMV DNA polymerase gene that is not affected by known drug resistance mutations. All 36 strains of CMV detected by culture or qualitative PCR in a population of lung transplant recipients were detected. The assay detected 1 to 10 copies of CMV plasmid DNA. The analytic sensitivity was not affected by the presence of DNA from 10(6) human cells but was reduced approximately 10-fold by alkaline lysates of leukocyte preparations. CMV quantitation was linear over a range of 10(1) to 10(6) copies. The intraassay and interassay coefficients of variation were 29 and 40%. Human DNA was regularly detected in patient plasma samples, and the amount was increased by storage of blood at room temperature before plasma separation and by plasma separation techniques that allowed leukocyte contamination. Applied to whole blood, the assay provides a measurement of CMV DNA in relation to cellular content without a need for cell counting procedures. Applied to plasma, the assay can reveal artifactual increases in plasma CMV levels resulting from leukocyte contamination. Further study of the utility of this assay to monitor patient populations at risk for CMV disease is warranted. PMID- 12089252 TI - Sensitivity of the Procleix HIV-1/HCV assay for detection of human immunodeficiency virus type 1 and hepatitis C virus RNA in a high-risk population. AB - The Procleix HIV-1/HCV Assay is a high-throughput nucleic acid test for the simultaneous detection of human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV) RNA during blood donor screening. This study evaluated the clinical sensitivity of the Procleix assay and assessed the assay's ability to identify HIV-1- and HCV-infected individuals undetected by standard serologic tests. Plasma samples were obtained prospectively from 539 individuals at high risk for HIV-1 and HCV infection at seven clinics affiliated with Johns Hopkins University. Samples were tested in the Procleix HIV-1/HCV Assay and, if reactive, were then tested in the Procleix HIV-1 and HCV discriminatory assays to differentiate the source of viral RNA positivity. Of these 539 subjects, 287 (53.2%) tested reactive in the Procleix HIV-1/HCV Assay. In discriminatory assay testing, 12 of 287 subjects (4.2%) were reactive for HIV-1 RNA only, 260 (90.6%) were reactive for HCV RNA only, and 11 (3.8%) were coinfected with HIV-1 and HCV. The clinical sensitivity for samples tested neat was 100% for HIV-1 and 99.3% for HCV. Three subjects with Procleix HCV reactive/seronegative results seroconverted upon follow-up and were confirmed as Procleix HCV yield cases. The Procleix HIV 1/HCV Assay is a highly sensitive test that detects ongoing and early HIV-1 and HCV infection in a significant number of subjects at high risk for these diseases. Confirmation of Procleix yield cases upon follow-up demonstrated the ability of the Procleix HIV-1/HCV Assay to detect the presence of HIV-1 and HCV in blood earlier than standard serologic tests. PMID- 12089253 TI - Detection of methicillin-resistant Staphylococcus aureus and simultaneous confirmation by automated nucleic acid extraction and real-time PCR. AB - A molecular assay for the simultaneous detection of a Staphylococcus aureus specific gene and the mecA gene, responsible for the resistance to methicillin in staphylococci, was evaluated. The assay included an automated DNA extraction protocol conducted with a MagNA Pure instrument and real-time PCR conducted with a LightCycler instrument. The performance and robustness of the assay were evaluated for a suspension of methicillin-resistant S. aureus (MRSA) strain with a turbidity equivalent to a McFarland standard of 0.5, which was found to be the ideal working concentration. The specificity of the new molecular assay was tested with a panel of 30 gram-negative and gram-positive bacterial strains other than MRSA. No cross-reactivity was observed. In a clinical study, 109 isolates of MRSA were investigated. All clinical MRSA isolates gave positive results for the S. aureus-specific genomic target, and all but one were positive for the mecA gene. In conclusion, the new molecular assay was found to be quick, robust, and laborsaving, and it proved to be suitable for a routine molecular diagnostic laboratory. PMID- 12089254 TI - Detection and genotyping of human group A rotaviruses by oligonucleotide microarray hybridization. AB - A rapid and reliable method for the identification of five clinically relevant G genotypes (G1 to G4 and G9) of human rotaviruses based on oligonucleotide microarray hybridization has been developed. The genotype-specific oligonucleotides immobilized on the surface of glass slides were selected to bind to the multiple target regions within the VP7 gene that are highly conserved among individual rotavirus genotypes. Rotavirus cDNA was amplified in a PCR with primers common to all group A rotaviruses. A second round of nested PCR amplification was performed in the presence of indodicarbocyanine-dCTP and another pair of degenerate primers also broadly specific for all genotypes. The use of one primer containing 5'-biotin allowed us to prepare fluorescently labeled single-stranded hybridization probe by binding of another strand to magnetic beads. The identification of rotavirus genotype was based on hybridization with several individual genotype-specific oligonucleotides. This approach combines the high sensitivity of PCR with the selectivity of DNA-DNA hybridization. The specificity of oligonucleotide microchip hybridization was evaluated by testing 20 coded rotavirus isolates from different geographic areas for which genotypes were previously determined by conventional methods. Analysis of the coded specimens showed that this microarray-based method is capable of unambiguous identification of all rotavirus strains. Because of the presence of random mutations, each individual virus isolate produced a unique hybridization pattern capable of distinguishing different isolates of the same genotype and, therefore, subgenotype differentiation. This strain information indicates one of several advantages that microarray technology has over conventional PCR techniques. PMID- 12089255 TI - Highly sensitive multiplex assay for detection of human immunodeficiency virus type 1 and hepatitis C virus RNA. AB - Various nucleic acid assays have been developed and implemented for diagnostics and therapeutic monitoring of human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV) infections. The high-throughput, semiautomated assays described here were developed to provide a method suitable for screening plasma specimens for the presence of HIV-1 and HCV RNAs. Three assays were developed: a multiplex HIV-1/HCV assay for simultaneous detection of HIV-1 and HCV, and discriminatory assays for specific detection of HIV-1 and HCV. The assay systems utilize three proprietary technologies: (i) target capture-based sample preparation, (ii) transcription-mediated amplification (TMA), and (iii) hybridization protection assay (HPA). An internal control is incorporated into each reaction to control for every step of the assay and identify random false negative reactions. The assays demonstrated a sensitivity of at least 100 copies/ml for each target, and they detected with similar sensitivity all major variants of HCV and HIV-1, including HIV-1 group O strains. Assay sensitivity for one virus was not affected by the presence of the other. The specificity of these TMA-driven assays was >or=99.5% in both normal donor specimens and plasma containing potentially interfering substances or other blood-borne pathogens. Statistical receiver operating characteristic plots of 1 - specificity versus sensitivity data determined very wide analyte cutoff values for each assay at the point at which the assay specificity and sensitivity were both >or=99.5%. The sensitivity, specificity, and throughput capability predict that these assays will be valuable for large-volume plasma screening, either in a blood bank setting or in other diagnostic applications. PMID- 12089256 TI - Occurrence of a multidrug-resistant Pseudomonas aeruginosa clone in different hospitals in Rio de Janeiro, Brazil. AB - Multidrug-resistant Pseudomonas aeruginosa nosocomial infections are increasingly recognized worldwide. The existence of metallo-beta-lactamase- and extended spectrum beta-lactamase-producing isolates exhibiting resistance to most beta lactam antimicrobial agents greatly complicates the clinical management of patients infected with such isolates. Since 1998, P. aeruginosa isolates resistant to all commercially available antimicrobial agents have been detected at a university-affiliated public hospital in Rio de Janeiro, Brazil. The present study was designed to characterize the antimicrobial resistance profiles and the genetic diversity of the P. aeruginosa strains isolated at this hospital and four private hospitals in Rio de Janeiro. Between April 1999 and March 2000, 200 consecutive isolates were obtained and analyzed for antimicrobial resistance. The genetic diversity of a selected number of them was evaluated by pulsed-field gel electrophoresis and PCR with the ERIC-2 primer. A predominant genotype, designated genotype A, was identified among isolates from four of the five hospitals evaluated. Eighty-four ceftazidime-resistant isolates were evaluated for metallo-beta-lactamase production, which was detected in 20 (91%) of 22 genotype A isolates and 11 (18%) of 62 isolates belonging to other genotypes (P < 0.05). Two metallo-beta-lactamase-producing genotype A isolates also produced an extended-spectrum beta-lactamase. The occurrence of multidrug-resistant P. aeruginosa strains belonging to a unique genotype in different hospitals in Rio de Janeiro underscores the importance of the contribution of a single clone to the increase in the incidence of multidrug-resistant P. aeruginosa nosocomial infections. PMID- 12089257 TI - Histologic and genotypic characterization of a novel Mycobacterium species found in three cats. AB - Three cases of feline atypical mycobacteriosis from different geographical regions in North America were characterized by large clusters of filamentous bacteria visible on hematoxylin-and-eosin-stained tissue sections. PCR amplification demonstrated the presence of Mycobacterium-specific nucleic acid in samples of skin lesions from these cases. PCR-assisted cloning and DNA sequence analysis of a 541-bp length of the Mycobacterium 16S rRNA gene generated DNA sequences which were >95% identical, suggesting that the three isolates were closely related. Two of the sequences were 99% identical and may represent the same species. Alignment with comparable 16S rRNA gene sequences from 66 Mycobacterium species and partially characterized isolates highlighted similarities (>94%) with Mycobacterium bohemicum, Mycobacterium haemophilum, Mycobacterium ulcerans, Mycobacterium avium subsp. avium, and isolate IWGMT 90242. Parsimony analysis of sequence data suggested relatedness to M. leprae. Significant molecular genetic and pathobiological differences between these three similar isolates and other known species of mycobacteria suggested that the organisms may not have been described previously and that these cases may represent a new form of mycobacterial disease in cats. We suggest the term "Mycobacterium visibilis" to describe the organism from which the two nearly identical sequences were obtained. PMID- 12089258 TI - Seroprevalence of Babesia infections in humans exposed to ticks in midwestern Germany. AB - Babesiosis is considered to be an emerging tick-borne disease in humans worldwide. However, most studies on the epidemiology of human babesiosis to date have been carried out in North America, and there is little knowledge on the prevalence of infection and frequency of disease in other areas. The aim of this study was to investigate the prevalence of Babesia infections in a human population in Germany. A total of 467 sera collected between May and October 1999 from individuals living in the Rhein-Main area were tested for the presence of immunoglobulin G (IgG) and IgM antibodies to antigens of Babesia microti and Babesia divergens by indirect fluorescent-antibody (IFA) tests. These sera were derived from 84 Lyme borreliosis patients suffering from erythema migrans, 60 asymptomatic individuals with positive borreliosis serology, and 81 individuals with a history of tick bite. Cutoff values for discrimination between seronegative and seropositive results in the IFA tests were determined using sera from 120 healthy blood donors and 122 patients suffering from conditions other than tick-borne diseases (malaria, n = 40; toxoplasmosis, n = 22; syphilis, n = 20; Epstein-Barr virus infection, n = 20; and presence of antinuclear antibodies, n = 20). The overall specificities of the IFA tests for B. microti and B. divergens were estimated to be >or=97.5%. Positive IgG reactivity against B. microti antigen (titer, >or=1:64) or B. divergens antigen (titer, >or=1:128) was detected significantly more often (P < 0.05) in the group of patients exposed to ticks (26 of 225 individuals; 11.5%) than in the group of healthy blood donors (2 of 120 individuals; 1.7%). IgG antibody titers of >or=1:256 against at least one of the babesial antigens were found significantly more often (P < 0.05) in patients exposed to ticks (9 of 225) than in the control groups (1 of 242). In the human population investigated here, the overall seroprevalences for B. microti and B. divergens were 5.4% (25 of 467) and 3.6% (17 of 467), respectively. The results obtained here provide evidence for concurrent infections with Borrelia burgdorferi and Babesia species in humans exposed to ticks in midwestern Germany. They also suggest that infections with Babesia species in the German human population are more frequent than believed previously and should be considered in the differential diagnosis of febrile illness occurring after exposure to ticks or blood transfusions, in particular in immunocompromised patients. PMID- 12089259 TI - Minimizing the workup of blood culture contaminants: implementation and evaluation of a laboratory-based algorithm. AB - An algorithm was implemented in the clinical microbiology laboratory to assess the clinical significance of organisms that are often considered contaminants (coagulase-negative staphylococci, aerobic and anaerobic diphtheroids, Micrococcus spp., Bacillus spp., and viridans group streptococci) when isolated from blood cultures. From 25 August 1999 through 30 April 2000, 12,374 blood cultures were submitted to the University of Iowa Clinical Microbiology Laboratory. Potential contaminants were recovered from 495 of 1,040 positive blood cultures. If one or more additional blood cultures were obtained within +/ 48 h and all were negative, the isolate was considered a contaminant. Antimicrobial susceptibility testing (AST) of these probable contaminants was not performed unless requested. If no additional blood cultures were submitted or there were additional positive blood cultures (within +/-48 h), a pathology resident gathered patient clinical information and made a judgment regarding the isolate's significance. To evaluate the accuracy of these algorithm-based assignments, a nurse epidemiologist in approximately 60% of the cases performed a retrospective chart review. Agreement between the findings of the retrospective chart review and the automatic classification of the isolates with additional negative blood cultures as probable contaminants occurred among 85.8% of 225 isolates. In response to physician requests, AST had been performed on 15 of the 32 isolates with additional negative cultures considered significant by retrospective chart review. Agreement of pathology resident assignment with the retrospective chart review occurred among 74.6% of 71 isolates. The laboratory based algorithm provided an acceptably accurate means for assessing the clinical significance of potential contaminants recovered from blood cultures. PMID- 12089260 TI - Real-time PCR for quantification of human herpesvirus 6 DNA from lymph nodes and saliva. AB - A real-time quantitative PCR assay has been developed to measure human herpesvirus 6 (HHV-6) DNA in biological specimens. The assay sensitivity was 10 copies of DNA per well, with a linear dynamic range of 10 to 10(7) copies of HHV 6 DNA. Intra- and interassay variations were, respectively, 0.88 and 0.8% for samples containing 10(2) DNA copies, 0.99 and 0.96% for samples containing 10(4) copies, and 0.76 and 0.9% for samples containing 10(6) copies. Among 34 saliva samples from healthy subjects, 26 were found to contain HHV-6 DNA (76.5%; median, 23,870 copies/ml), and following a single freeze-thaw cycle, 25 of the same samples were found to be positive for HHV-6 DNA, although at a statistically significantly lower concentration (median, 3,497 copies/ml). The assay enabled detection of HHV-6 DNA in lymph node biopsies from patients with Hodgkin's disease (HD) (13 of 37 patients [35.1%]), B-cell neoplasms (8 of 36 patients [22.2%]), and T- or NK-cell neoplasms (3 of 13 patients [23.1%]), with concentrations ranging from 100 to 864,640 HHV-6 copies per microg of DNA (HHV-6B being found in every case except two). All HD patients infected with HHV-6 presented clinically with the nodular sclerosis subtype of HD. The real-time quantitative PCR assay developed here was simple to perform and was sensitive over a wide range of HHV-6 concentrations. It therefore appears to be of potential value in clinical investigation or diagnosis of HHV-6 infection. PMID- 12089261 TI - Clostridium difficile genotyping based on slpA variable region in S-layer gene sequence: an alternative to serotyping. AB - Recent investigations of Clostridium difficile cell wall components have revealed the presence of an S-layer encoded by the slpA gene. The aim of this study was to determine whether slpA genotyping can be used as an alternative to serotyping. The variable regions of slpA were amplified by PCR from serogroup reference strains and various clinical isolates chosen randomly. Amplified products were analyzed after restriction enzyme digestion and DNA sequencing. The sequences of the variable region of the SlpA protein were found to be strictly identical within a given serogroup but divergent between serogroups. These preliminary results suggest that PCR-restriction fragment length polymorphism, in conjunction with DNA sequencing of the slpA variable region, could constitute an alternative typing method for determining C. difficile serotypes. PMID- 12089262 TI - Cross-reactivity among several recombinant calicivirus virus-like particles (VLPs) with monoclonal antibodies obtained from mice immunized orally with one type of VLP. AB - Human caliciviruses (HuCVs) are classified into the Norwalk-like viruses (NLV) and Sapporo-like viruses (SLV) as genera within the family CALICIVIRIDAE: The NLV genus is further classified into genogroups I and II, based on sequence similarities. To study the antigenic determinants on the HuCV capsid protein and develop new diagnostic tools for field samples, we established and characterized monoclonal antibodies (MAbs) against baculovirus-expressed recombinant HuCV virus like particles (VLPs). Hybrid clones producing MAbs were obtained from cultures of PAI myeloma cells fused with spleen or mesenteric lymph node cells from mice immunized orally with either a single type of recombinant Norwalk virus (rNV), Kashiwa 47 virus (rKAV), Snow Mountain agent (rSMA), or Sapporo virus (rSV) VLP or with mixtures of two types of VLPs from different genogroups. Twenty MAbs, obtained as mouse ascites, were characterized and classified into six groups according to their enzyme-linked immunosorbent assay (ELISA) and Western blotting (WB) cross-reactivity patterns to VLPs. Five groups of MAbs reacted by both WB and ELISA and were classified as follows: common cross-reactive MAbs for four genogroup I and six genogroup II VLPs (group A), genogroup I-specific MAbs (group B), genogroup II-specific MAbs (group C), and strain-specific MAbs (groups D and E). One MAb group (group F) reacted only by ELISA. The group A MAbs, which showed broad cross-reactivity with VLPs of both NLV genogroups, were obtained from mice immunized orally with a single type of VLP (either rNV or rKAV). Two MAbs, which were obtained from mice immunized with rSV, reacted with rSV but not with any NLV VLP. These are the first MAbs to be reported for any SLV. These strain-, genogroup-, and genus-reactive MAbs will be useful tools for further study of the antigenic and structural topography of the HuCV virion and for diagnostic assays for HuCVs. PMID- 12089263 TI - Detection of Tropheryma whipplei DNA in feces by PCR using a target capture method. AB - Whipple's disease is a rare multisystemic bacterial infection with variable clinical manifestations. For decades, the laboratory diagnosis was based on the demonstration of periodic acid Schiff-positive inclusions in macrophages of gastrointestinal biopsies. PCR has improved the diagnosis of Whipple's disease due to its increased sensitivity compared to histopathological analysis. To avoid invasive procedures for taking specimens, we have investigated the possibility of detecting Tropheryma whipplei DNA in feces rather than in biopsies or gastric aspirate of patients with and without Whipple's disease. Total bacterial DNA was isolated from stool specimens using Qiagen columns followed by a T. whipplei specific hybridization step with a biotinylated capture probe and streptavidin coated magnetic particles. The captured DNA was then amplified using the same seminested PCR targeting the 16S rRNA gene of the organism that had been applied to other specimens without capturing. For five of eight patients with Whipple's disease, duodenal biopsies and stool samples were PCR positive, whereas for the three other patients, both specimens were PCR negative. Of 84 patients without Whipple's disease, 75 tested negative in the duodenal biopsy and in the stool sample. For four, both specimens were positive. Five patients tested positive in the stool sample but not in the biopsy. However, for three of these five patients, the gastric aspirate had been PCR positive, indicating that the stool PCR result was true rather than false positive. Compared to PCR from duodenal biopsies, stool PCR has a sensitivity of 100% and a specificity of 97.3%. Additionally, 15 PCR-positive and 22 PCR-negative stool samples were extracted using the Invisorb Spin Stool DNA kit. The simplified stool extraction showed 93.3% sensitivity and 95.5% specificity compared to the target capture method. We conclude that PCR with stool specimens with either extraction method is a sensitive and specific diagnostic tool for the detection of T. whipplei DNA and one not requiring invasive sampling procedures. PMID- 12089264 TI - Comparison of an assay using signal amplification of the heat-dissociated p24 antigen with the Roche Monitor human immunodeficiency virus RNA assay. AB - We compared an assay using signal amplification of a heat-dissociated p24 antigen (HDAg) with the Roche Monitor human immunodeficiency virus (HIV) RNA assay. The two assays gave comparable results when 130 specimens from 130 patients were tested (r = 0.60, P < 0.0001). The HDAg assay was almost as sensitive (85%) as the Roche HIV RNA kit (95%), just as specific (25 negative results from 25 HIV seronegative volunteers [100%]), less variable (mean log standard deviation of 0.07 compared to 0.11 when eight specimens were tested three or four times), and less expensive (reagent and labor costs, $8 versus $75). The assay appeared to be useful for monitoring established patients (n = 17) and identifying seroconverters (n = 4). HIV subtypes A to F were all recognized. This assay should be useful for monitoring patients in resource-poor countries and for monitoring vaccine recipients. PMID- 12089265 TI - Evaluation of 15 motility media and a direct microscopic method for detection of motility in enterococci. AB - Isolation of motile, vanC enterococci has yet to be a major infection control concern; however, rapid detection still is important. We evaluated 15 motility media from three manufacturers and a 2-h direct microscopic method for accurate detection of 89 enterococcal strains, including 72 vanC enterococcal strains. Resistance genes were confirmed by a multiplex PCR method with the vanC gene detected in all motile enterococci. Motility in the 72 vanC enterococci was detected at 30 degrees C within 72 h in BD Biosciences motility nitrate medium and in Remel motility B medium, motility B medium supplemented with methyl-alpha D-glucopyranoside (investigational), motility S medium, motility test medium, and motility test medium with tetrazolium indicator. Motility was also observed for all vanC enterococci with the 2-h direct (30 degrees C incubation) microscopic detection method. All Enterococcus faecalis and Enterococcus faecium isolates were observed to be nonmotile in all media and by the direct microscopic method. Since differences between the various motility media tested were observed, the medium used for detection of enterococcal motility must be selected carefully. PMID- 12089266 TI - Performance of CHROMagar selective medium and oxacillin resistance screening agar base for identifying Staphylococcus aureus and detecting methicillin resistance. AB - Two new selective media, oxacillin resistance screening agar base (ORSAB) and CHROMagar Staph aureus (CSA), were evaluated for identification of Staphylococcus aureus and for screening of methicillin resistance by addition of antimicrobial agents to these media. A well-defined collection consisting of 1,140 staphylococci was used. A total of 624 were S. aureus, of which 358 were methicillin susceptible and 266 were methicillin resistant, and 516 were coagulase-negative staphylococci. The methicillin-resistant S. aureus (MRSA) strains were selected based on the results of phage typing; 247 different types were included in the analysis. For identification of S. aureus, both media performed better after 24 h than after 48 h. The sensitivities at 24 h were comparable (CSA, 98.6%; ORSAB, 97.1%), but the specificity of CSA was significantly higher (CSA, 97.1%; ORSAB, 92.1%). For screening of methicillin resistance, antibiotic supplements were added to both media. The sensitivity was lower after 24 h (CSA, 58.6%; ORSAB, 84.2%) and increased significantly after 48 h (CSA, 77.5%; ORSAB, 91.4%). At both time intervals ORSAB was significantly more sensitive than CSA. However, the specificities of both media were high after 24 h (CSA, 99.1%; ORSAB, 98.3%) and decreased significantly after 48 h of incubation (CSA, 94.7%; ORSAB, 95.5%). In conclusion, for identification of S. aureus, CSA is more accurate than ORSAB because of a significantly higher specificity. For screening of MRSA, ORSAB performs better than CSA, but the usefulness in clinical practice is limited because a significant number of strains are not detected. PMID- 12089267 TI - Seminested PCR for diagnosis of candidemia: comparison with culture, antigen detection, and biochemical methods for species identification. AB - The rapid detection and identification of Candida species in clinical laboratories are extremely important for the management of patients with hematogenous candidiasis. The presently available culture and biochemical methods for detection and species identification of Candida are time-consuming and lack the required sensitivity and specificity. In this study, we have established a seminested PCR (snPCR) using universal and species-specific primers for detection of Candida species in serum specimens. The universal outer primers amplified the 3' end of 5.8S ribosomal DNA (rDNA) and the 5' end of 28S rDNA, including the internally transcribed spacer 2 (ITS2), generating 350- to 410-bp fragments from the four commonly encountered Candida species, viz., C. albicans, C. tropicalis, C. glabrata, and C. parapsilosis. The species-specific primers, complementary to unique sequences within the ITS2 of each test species, amplified species-specific DNA in the reamplification step of the snPCR. The sensitivity of Candida detection by snPCR in spiked serum specimens was close to 1 organism/ml. Evaluation of snPCR for specific identification of Candida species with 76 clinical Candida isolates showed 99% concordant results with the Vitek and/or ID32C yeast identification system. Further evaluation of snPCR for detection of Candida species in sera from culture-proven (n = 12), suspected (n = 16), and superficially colonized (n = 10) patients and healthy subjects (n = 12) showed that snPCR results were consistently negative with sera from healthy individuals and colonized patients. In culture-proven candidemia patients, the snPCR results were in full agreement with blood culture results with respect to both positivity and species identity. In addition, snPCR detected candidemia due to two Candida species in five patients, compared to three by blood culture. In the category of suspected candidemia with negative blood cultures for Candida, nine patients (56%) were positive by snPCR; two of them had dual infection with C. albicans and either C. tropicalis or C. glabrata. In conclusion, the snPCR developed in this study is specific and more sensitive than culture for the detection of Candida species in serum specimens. Moreover, the improved detection of cases of candidemia caused by more than one Candida species is an additional advantage. PMID- 12089269 TI - Genetic diversity of clinical and environmental strains of Salmonella enterica serotype Weltevreden isolated in Malaysia. AB - The incidence of food-borne salmonellosis due to Salmonella enterica serotype Weltevreden is reported to be on the increase in Malaysia. The pulsed-field gel electrophoresis (PFGE) subtyping method was used to assess the extent of genetic diversity and clonality of Salmonella serotype Weltevreden strains from humans and the environment. PFGE of XbaI-digested chromosomal DNA from 95 strains of Salmonella serotype Weltevreden gave 39 distinct profiles with a wide range of Dice coefficients (0.27 to 1.00), indicating that PFGE is very discriminative and that multiple clones of Salmonella serotype Weltevreden exist among clinical and environmental isolates. Strains of one dominant pulsotype (pulsotype X1/X2) appeared to be endemic in this region, as they were consistently recovered from humans with salmonellosis between 1996 and 2001 and from raw vegetables. In addition, the sharing of similar PFGE profiles among isolates from humans, vegetables, and beef provides indirect evidence of the possible transmission of salmonellosis from contaminated raw vegetables and meat to humans. Furthermore, the recurrence of PFGE profile X21 among isolates found in samples of vegetables from one wet market indicated the persistence of this clone. The environment in the wet markets may represent a major source of cross-contamination of vegetables with Salmonella serotype Weltevreden. Antibiotic sensitivity tests showed that the clinical isolates of Salmonella serotype Weltevreden remained drug sensitive but that the vegetable isolates were resistant to at least two antibiotics. To the best of our knowledge, this is the first study to compare clinical and environmental isolates of Salmonella serotype Weltevreden in Malaysia. PMID- 12089268 TI - Phenotypic and genotypic characterization of serologically atypical strains of Shigella flexneri type 4 isolated in Dhaka, Bangladesh. AB - Twenty-one atypical Shigella flexneri type 4 strains isolated from patients attending the Dhaka treatment center of the International Centre for Diarrhoeal Disease Research, Bangladesh, were extensively characterized and compared with S. flexneri serotypes 4a and 4b. The atypical strains agglutinated only with the type antigen factor 4 and did not agglutinate with any group factors, thereby excluding their characterization into serotype 4a or 4b. Of the 21 strains, 85.7% did not ferment mannitol but were able to ferment most of the sugars, whereas the remaining 14.3% strains fermented mannitol but were unable to ferment most of the sugars. Most of the strains were resistant to ampicillin, tetracycline, and trimethoprim-sulfomethoxazole. All of the strains harbored the 140-MDa plasmid, had the ipaH gene, had the sen gene (encoding Shigella enterotoxin 2), had the ability to bind Congo red, and were positive for keratoconjunctivitis in the guinea pig eye, attesting their invasive properties. All of the strains contained a middle-range plasmid (35 to 62 MDa) as well as a number of stable small plasmids, yielding mainly two plasmid profiles which were different from those of 4a and 4b strains. Conjugation and curing experiments suggested that the middle range plasmids harbored a self-transferable multiple antibiotic resistance marker. Pulsed-field gel electrophoresis analysis of all of the tested strains yielded two types with numerous subtypes, whereas ribotyping yielded only two types which were completely different from those of types 4a and 4b. This study concluded that two different clones of atypical S. flexneri type 4 exist and strongly suggests that these are new subserotypes of S. flexneri that await further serological classification. PMID- 12089270 TI - VIDAS test for avidity of Toxoplasma-specific immunoglobulin G for confirmatory testing of pregnant women. AB - Because congenital toxoplasmosis is almost solely the result of maternal infection acquired during gestation, it is critical to determine whether infection during pregnancy has occurred. In the United States, definitive diagnosis of the acute infection and the time of its occurrence have been compromised by a lack of systematic screening and the fact that only a single serum sample is submitted for testing. In studies in Europe, and depending on the method used, the demonstration of high-avidity immunoglobulin G (IgG) toxoplasma antibodies has been shown to exclude infection having occurred in the first 3 to 5 months of pregnancy. We investigated the usefulness of determining the avidity of IgG toxoplasma antibodies with a VIDAS kit (herein referred to as the VIDAS Toxo-IgG avidity kit, the VIDAS kit essentially rules out acute infection having occurred within the 4 prior months) in the setting of a reference serology laboratory in the United States. Sera (132 samples) from 132 women in the first 16 weeks of pregnancy were chosen because at least one test in the toxoplasma serological profile (TSP) suggested or was equivocal for a recently acquired infection. High-avidity antibodies were demonstrated in 75% of 99 sera positive with the IgM enzyme-linked immunosorbent assay (ELISA) and 31.3% of 16 sera with acute TSP results. A significant percentage of sera with equivocal results wtih the IgM ELISA or TSP also had high-avidity test results. Of 39 women for whom treatment with spiramycin had been suggested to attempt to prevent congenital transmission, 19 (48.7%) had high-avidity antibodies. These findings highlight the value of the VIDAS IgG avidity kit when used in combination with the TSP to exclude recent infection, especially when only a single serum sample is available. PMID- 12089271 TI - Detection of isoniazid-resistant Mycobacterium tuberculosis strains by a multiplex allele-specific PCR assay targeting katG codon 315 variation. AB - We describe a simple multiplex allele-specific (MAS)-PCR assay to detect mutations in the second base of the katG gene codon 315, including AGC-->ACC and ACA (Ser-->Thr) substitutions that confer resistance to isoniazid (INH) in Mycobacterium tuberculosis clinical isolates. The 315 ACC allele is found in the majority of Inh(r) strains worldwide, especially in areas with a high incidence of tuberculosis. The 315 ACA allele is characteristic of the New York City multidrug-resistant (MDR) strain W and its progenies in the United States. The mutations in katG315 are revealed depending on the presence or absence of an indicative fragment amplified from the wild-type allele of this codon. Initially optimized on the purified DNA samples, the assay was then tested on crude cell lysates and auramine-stained sputum slide preparations with the same reproducibility and interpretability of profiles generated by agarose gel electrophoresis. The MAS-PCR assay can be used for the detection of resistance to INH in clinical laboratories in regions with a high prevalence of MDR M. tuberculosis strains. PMID- 12089272 TI - Helicobacter marmotae sp. nov. isolated from livers of woodchucks and intestines of cats. AB - Woodchucks (Marmota monax) have a high incidence of hepatocellular carcinoma (HCC) associated with chronic infection with woodchuck hepatitis virus (WHV) and serve as a model of hepatitis B virus-associated HCC in humans. Helicobacter hepaticus, an enterohepatic helicobacter in mice, is known to cause hepatocellular adenomas and carcinomas in susceptible mouse strains. In long-term chemical bioassays conducted with B6C3F(1) mice, H. hepaticus has been regarded as a confounding factor because of its tumor-promoting activity. In order to determine if woodchucks harbor a Helicobacter sp. that might play a role in potentiating hepatic inflammation or neoplasia, a study was undertaken to determine whether woodchucks' livers were infected with a Helicobacter sp. Frozen liver samples from 20 (17 WHV-infected and 3 noninfected) woodchucks, 10 with WHV associated hepatic tumors and 10 without tumors, were cultured by microaerobic techniques and analyzed by using genus- and species-specific helicobacter PCR primers. A 1,200-bp Helicobacter sp.-specific sequence was amplified from 14 liver samples. Southern hybridization confirmed the specific identity of the PCR products. Nine of the 10 livers with tumors had positive Helicobacter sp. identified by PCR, whereas 5 of the 10 livers without tumors were positive. By use of 16S rRNA species-specific primers for H. marmotae, two additional liver samples from the nontumor group had positive PCR amplicons confirmed by Southern hybridization. A urease-, catalase-, and oxidase-positive bacterium was isolated from one liver sample from a liver tumor-positive woodchuck. By 16S rRNA analysis and biochemical and phenotypic characteristics, the organism was classified as a novel Helicobacter sp. Subsequently, four additional bacterial strains isolated from feces of cats and characterized by biochemical, phenotypic, and 16S rRNA analysis were determined to be identical to the woodchuck isolate. We propose the name Helicobacter marmotae sp. nov. for these organisms. Further studies are required to ascertain if this novel Helicobacter sp. plays a tumor promotion role in hepadnavirus-associated tumors in woodchucks or causes enterohepatic disease in cats. PMID- 12089273 TI - Nosocomial spread of a Staphylococcus capitis strain with heteroresistance to vancomycin in a neonatal intensive care unit. AB - A premature infant in a neonatal intensive care unit (NICU) developed a bloodstream infection caused by coagulase-negative staphylococci (CoNS) sensitive to vancomycin. The infection persisted for 3 weeks, despite therapy with vancomycin and replacement of all intravenous catheters. The neonate died due to necrotizing enterocolitis which developed during the ongoing sepsis. We screened this strain and 216 other strains of CoNS from cultures of blood obtained from neonates between 1997 and 2000 for heteroresistance to vancomycin. Forty-eight isolates, including the strain that caused ongoing sepsis, proved heteroresistant. All isolates were identified as Staphylococcus capitis and were identical, just as their resistant stable subcolonies were, when they were genetically fingerprinted by amplified-fragment length polymorphism analysis. The heteroresistant phenotype of this endemic strain was confirmed by population analysis. We conclude that heteroresistance to vancomycin occurs in S. capitis and might be the cause of therapeutic failures in NICUs. Moreover, heteroresistant strains can become endemic in such units. PMID- 12089274 TI - Use of a fragment of glycoprotein G-2 produced in the baculovirus expression system for detecting herpes simplex virus type 2-specific antibodies. AB - Fragments of glycoprotein G (gG-2(281-594His)), comprising residues 281 to 594 of herpes simplex virus type 2 (HSV-2), glycoprotein G of HSV-1 (gG-1(t26-189His)), and glycoprotein D of HSV-1 (gD-1(1-313)), were expressed in the baculovirus expression system to develop an assay for the detection of HSV-1 and HSV-2 type specific antibodies. The expression of the gG-1(t26-189His) and gG-2(281-594His) fragments was analyzed by Western blotting using monoclonal antibodies LP10 and AP1, respectively. The molecular masses of the major products of gG-1(t26-189His) and the fragment of gG-2(281-594His) were 36 to 39 kDa and 64 to 72 kDa, respectively. Human sera positive for HSV-1 reacted with gG-1(t26-189His), sera positive for HSV-2 reacted with the gG-2(281-594His) fragment, and sera positive for both types reacted with gG-1(t26-189His) and gG-2(281-594His) in Western blotting. The human sera recognized polypeptides of gG-2(281-594His) with molecular masses of 57 to 67 and 120 to 150 kDa and additional faint bands of 21, 29, and 45 kDa. The recombinant gG-1(t26-189His) and the recombinant gG-2(281 594His) fragment were used as type-specific antigens for the detection of HSV-1- and HSV-2-specific antibody responses in human sera, respectively. As type-common antigens, an extract of HSV-1-infected Vero cells and recombinant gD-1(1-313) were used. An enzyme-linked immunosorbent assay to detect type-specific antibodies was developed, and the sensitivity and specificity were evaluated by comparison with commercial tests by using sera obtained from different sources. The sensitivity and specificity were 91.5 and 95.5%, respectively, compared to the Gull assay. The gG-2(281-594His) fragment can be obtained in relatively large quantities at low cost. PMID- 12089275 TI - Detection of Epstein-Barr virus genomes in peripheral blood B cells from solid organ transplant recipients by fluorescence in situ hybridization. AB - Resolution of Epstein-Barr Virus (EBV) infection in pediatric solid-organ transplant recipients often leads to an asymptomatic carrier state characterized by a persistently elevated circulating EBV load that is 2 to 4 orders of magnitude greater than the load typical of healthy latently infected individuals. Elevated EBV loads in immunosuppressed individuals are associated with an increased risk for development of posttransplant lymphoproliferative disease. We have performed fluorescence in situ hybridization (FISH) studies with peripheral blood B cells from carriers of persistent EBV loads in order to directly quantitate the number of EBV genomes per infected cell. Patients were assigned to two groups on the basis of the level of the persistent load (low-load carriers, 8 to 200 genomes/10(5) peripheral blood lymphocytes; high-load carriers, >200 genomes/10(5) peripheral blood lymphocytes). FISH analysis revealed that the low load carriers predominantly had circulating virus-infected cells harboring one or two genome copies/cell. High-load carriers also had cells harboring one or two genome copies/cell; in addition, however, they carried a distinct population of cells with high numbers of viral genome copies. The increased viral loads correlated with an increase in the frequency of cells containing high numbers of viral genomes. We conclude that low-load carriers possess EBV-infected cells that are in a state similar to normal latency, whereas high-load carriers possess two populations of virus-positive B cells, one of which carries an increased number of viral genomes per cell and is not typical of normal latency. PMID- 12089276 TI - Molecular peculiarities of the lytA gene isolated from clinical pneumococcal strains that are bile insoluble. AB - The autolytic LytA amidase from 12 bile (deoxycholate)-insoluble streptococcal isolates (formerly classified as atypical Streptococcus pneumoniae) showing different antibiotic resistance patterns was studied. These atypical strains, which autolyze at the end of the stationary phase of growth, contain highly divergent lytA alleles (pairwise evolutionary distances of about 20%) compared to the lytA alleles of typical pneumococci. The atypical LytA amidases exhibit a peculiar deletion of two amino acids responsible for cell wall anchoring in the carboxy-terminal domain and have a reduced specific activity. These enzymes were inhibited by 1% deoxycholate but were activated by 1% Triton X-100, a detergent that could be used as an alternative diagnostic test for this kind of strain. Preparation of functional chimeric enzymes, PCR mutagenesis, and gene replacements demonstrated that the characteristic bile insolubility of these atypical strains was due to their peculiar carboxy-terminal domain and that the 2 amino-acid deletion was responsible for the inhibitory effect of deoxycholate. However, the deletion alone did not affect the specific activity of LytA. A detailed characterization of the genes encoding the 16S rRNA and SodA together with multilocus sequence typing indicated that the strains studied here are not a single clone and, although they cannot be strictly classified as typical pneumococci, they represent a quite diverse pool of organisms closely related to S. pneumoniae. The clinical importance of these findings is underlined by the role of the lytA gene in shaping the course of pneumococcal diseases. This study can also contribute to solving diagnostic problems and to understanding the evolution and pathogenic potential of species of the Streptococcus mitis group. PMID- 12089277 TI - Real-time fluorescence PCR assays for detection and characterization of Shiga toxin, intimin, and enterohemolysin genes from Shiga toxin-producing Escherichia coli. AB - PCR assays have proved useful for detecting and characterizing Shiga toxin producing Escherichia coli (STEC). Recent advances in PCR technology have facilitated the development of real-time fluorescence PCR assays with greatly reduced amplification times and improved methods for the detection of amplified target sequences. We developed and evaluated two such assays for the LightCycler instrument: one that simultaneously detects the genes for Shiga toxins 1 and 2 (stx(1) and stx(2)) and another that simultaneously detects the genes for intimin (eae) and enterohemolysin (E-hly). Amplification and sequence-specific detection of the two target genes were completed within 60 min. Findings from the testing of 431 STEC isolates of human and animal origin, 73 isolates of E. coli negative for stx genes, and 118 isolates of other bacterial species with the LightCycler PCR (LC-PCR) assays were compared with those obtained by conventional block cycler PCR analysis. The sensitivities and specificities of the LC-PCR assays were each 100% for the stx(1), eae, and E-hly genes and 96 and 100%, respectively, for the stx(2) gene. No stx(2) genes were detected from 10 stx(2f) positive isolates because of significant nucleotide differences in their primer annealing regions. Melting curve analyses of the amplified Shiga toxin genes revealed sequence variation within each of the tested genes that correlated with described and novel gene variants. The performance characteristics of the LC-PCR assays, such as their speed, detection method, and the potential subtyping information available from melting curve analyses, make them attractive alternatives to block cycler PCR assays for detecting and characterizing STEC strains. PMID- 12089278 TI - Sensitive detection of HBsAg mutants by a gap ligase chain reaction assay. AB - Hepatitis B virus (HBV) surface gene variants have been associated with diagnostic escape and immune escape following vaccination. The most common mutation observed in these variants is a glycine-to-arginine substitution at amino acid 145 (G145R). In order to sensitively detect the presence of this mutant in serum, a new molecular detection system was developed; in this new system, a gap ligase chain reaction (gLCR) assay was coupled with electrochemiluminescence detection of reaction products. The gLCR assay could detect approximately 10 copies of mutant DNA and could discriminate low levels of mutant DNA in the presence of excess wild-type DNA. Detection of the G145R mutant in clinical specimens was evaluated by testing 56 suspect serum specimens. The G145R mutation was observed in 18 of 28 HBV-DNA-positive samples. The approximate percentage of mutant present in each specimen was calculated by comparison with a standard curve of an increasing ratio of mutant DNA to wild-type DNA. Most samples contained a very low percentage of mutant virus (approximately 5%), with an observed range of approximately 3 to 74%. The G145R mutation was most frequently observed in specimens producing a diagnostic anomaly or from transplant patients but was also observed in specimens from vaccinated individuals and specimens in which HBsAg diagnostic escape was suspected. Therefore, the gLCR assay is a sensitive and specific method for detection of G145R mutants, which could be modified to include the detection of other HBV mutants. PMID- 12089279 TI - Genetic diversity of the outer surface protein C gene of southern Borrelia isolates and its possible epidemiological, clinical, and pathogenetic implications. AB - The ospC genes of 20 southern Borrelia strains were sequenced. The strains consisted of B. burgdorferi sensu stricto, B. andersonii, B. bissettii, one undescribed genospecies, MI-8, and one probably new Borrelia species, TXW-1. A high degree of similarity exists between B. burgdorferi sensu stricto and B. bissettii and between B. bissettii and B. andersonii. Lateral transfers of the ospC gene probably occurred between B. burgdorferi sensu stricto and B. bissettii and between B. bissettii and B. andersonii. Internal gene recombination appears to occur among them. The highest degree of genetic diversity among them was observed in the two variable domains (V1 and V2), semivariable domain (SV), and the species-specific epitopes (between amino acids 28 and 31). Differences in ospC sequences among southern strains reflect diversity at the strain and genospecies levels. MI-8, which was recognized as an undescribed genospecies in our previous reports, remains distinguishable in our current analysis of ospC genes and is distinct from B. burgdorferi sensu stricto. Interestingly, another undescribed southern isolate, TXW-1, was not amplified under various PCR conditions. Compared to European B. burgdorferi sensu stricto strains, American B. burgdorferi sensu stricto strains show greater genetic heterogeneity. Southern B. burgdorferi sensu stricto, B. andersonii, and B. bissettii isolates were intermixed with each other in the phylogenetic trees. In the derived trees in our work, at least one southeastern strain of B. burgdorferi, MI-2, most closely aligns with a so-called invasive cluster that possesses many proven human invasive strains. Transmission experiments show that MI-2 and the strains in this group of southern spirochetes are able to infect mice and hamsters and that the typical vector of Lyme disease, Ixodes scapularis, can acquire the spirochetes from infected mammals. Currently, strain MI-2 appears to be the only southern isolate among the 20 we analyzed that clusters with an OspC invasive group and thus might be invasive for humans. PMID- 12089281 TI - Evaluation of the C6 peptide enzyme-linked immunosorbent assay for individuals vaccinated with the recombinant OspA vaccine. AB - The C6 enzyme-linked immunosorbent assay (ELISA), based on a peptide (C6) that reproduces the sequence of invariable region 6 of VlsE, the antigenic variation protein of Borrelia burgdorferi, has been shown to be a sensitive and specific test for the serologic diagnosis of Lyme disease. We now report that none of 29 uninfected individuals vaccinated with the recombinant OspA vaccine had an antibody response to the C6 peptide. The C6 peptide ELISA can be used to diagnose Lyme disease in patients who have received the OspA vaccine. PMID- 12089280 TI - Detection of duck hepatitis B virus DNA on filter paper by PCR and SYBR green dye based quantitative PCR. AB - Duck hepatitis B virus (DHBV) belongs to the Hepadnaviridae family, which includes human Hepatitis B virus (HBV) and Woodchuck hepatitis virus. It is widely distributed in wild and domestic ducks due to congenital transmission. HBV is a worldwide health problem, with carriers at risk of developing cirrhosis and liver cancer. Medical staff and scientists working with HBV must be vaccinated because of its contagious nature. DHBV is a safe surrogate for HBV because of their similarities. Collection of serum and blood samples on filter paper has been used to screen for metabolic disorders, genetic diseases, and viral infection and for evolutionary studies of the genome. In this study, DHBV from serum and blood dried on filters was detected by PCR. A 0.1-microl sample was sufficient for detection. The immobilization potential of filter papers for DHBV was examined, and the highest yield of PCR products was observed with Whatman paper. Dried serum was stable under different storage temperatures for 4 weeks, but the yields of PCR products decreased when the temperature was >or=4 degrees C. The optimal condition for storage was -70 degrees C. A newly developed quantitative PCR based on monitoring the amplification by measuring the increase in fluorescence caused by the binding of SYBR green dye to double-stranded products was applied herein. DHBV genomic DNA cloned in a plasmid was used for the generation of standard DHBV DNA for quantitative PCR. It validated results from PCR in terms of the copy number of DHBV particles. The specificity of PCR was demonstrated by melting curve analysis, and the differentiation of two DHBV isolates amplified from dried serum was demonstrated based on their melting temperatures determined by GC contents and sequence. It was easier and simpler than other PCR-based DNA techniques. The use of serum dried on filters allows samples from distant field for which cold storage and transportation are a problem to be mailed to the diagnostic laboratory. Samples can be archived for comparison and used as a source of DNA for cloning and sequencing. PMID- 12089282 TI - Differences between Staphylococcus aureus isolates from medical and nonmedical hospital personnel. AB - It is unclear whether the levels of Staphylococcus aureus colonization of hospital personnel with patient exposure are increased or whether personnel become colonized with more antibiotic-resistant strains. Differences in nasal and hand carriage of S. aureus between medical and nonmedical hospital personnel were examined. No differences in nasal carriage between the two groups were found; however, there was a trend that suggested differences in the rates of hand carriage of S. aureus (18% of nonmedical personnel and 10% of medical personnel). Medical personnel were colonized with more antibiotic-resistant isolates than nonmedical personnel (mean, 2.8 versus 2.1 isolates [P < 0.03]), and the strain profiles indicated that they tended to be more clonal in origin, suggesting that exposure to hospital isolates alters the colonization profile. PMID- 12089283 TI - Differentiation of porcine Brachyspira species by a novel nox PCR-based restriction fragment length polymorphism analysis. AB - A novel PCR-based restriction fragment length polymorphism analysis of the Brachyspira nox gene was developed. The restriction patterns for Brachyspira hyodysenteriae, B. pilosicoli, B. intermedia, B. murdochii, and B. innocens were highly distinct with two restriction endonucleases only. The assay proved to be user-friendly and robust. PMID- 12089284 TI - Transmission of Campylobacter hyointestinalis from a pig to a human. AB - We report on a case of human gastroenteritis caused by the pathogen Campylobacter hyointestinalis. Recurrent watery diarrhea and intermittent vomiting were the most significant symptoms of the previously healthy patient. Whole-cell protein electrophoresis and 16S rRNA gene sequencing were used to identify this Campylobacter species. Investigation of the patient's surroundings led to the recovery of a second C. hyointestinalis strain originating from porcine feces. Subsequent typing of the human and the porcine isolates by pulsed-field gel electrophoresis revealed similar macrorestriction profiles, indicating transmission of this pathogen. PMID- 12089285 TI - Prevalence of enterotoxigenic Escherichia coli strains harboring the longus pilus gene in Brazil. AB - The longus type IV pilus gene (lngA) was highly prevalent (32.8%) among Brazilian enterotoxigenic Escherichia coli strains producing both heat-labile and heat stable enterotoxins and bearing the CFA/I, CS1CS3, or CS6 antigen. Furthermore, lngA was more often found in strains isolated from children with diarrhea than in strains isolated from children without diarrhea. PMID- 12089286 TI - Quantitative stability of DNA after extended storage of clinical specimens as determined by real-time PCR. AB - Viral DNA stored for extended periods can be amplified by PCR. However, it is unknown whether stored specimens give accurate quantitative results by newer real time PCR techniques. We therefore compared herpes simplex virus DNA levels in specimens before and after 16 months of storage. The levels of viral DNA remained stable whether the DNA was stored as purified DNA or unextracted DNA in a whole specimen. PMID- 12089287 TI - Seroprevalence of antibodies that react with Anaplasma phagocytophila, the agent of human granulocytic ehrlichiosis, in different populations in Westchester County, New York. AB - We determined the frequencies of antibodies to Anaplasma phagocytophila, the agent of human granulocytic ehrlichiosis (HGE), in different groups of adults and children from Westchester County, New York. The groups included 159 adult blood donors and 215 children who were seronegative for Borrelia burgdorferi antibodies, 118 adult patients and 57 children who were seropositive for B. burgdorferi antibodies, and 42 adult patients with culture-confirmed erythema migrans. Eighteen (11.3%) of the blood donors and 11 (5.1%) of the B. burgdorferi seronegative children were found to have A. phagocytophila antibodies by indirect immunofluorescent-antibody assay (IFA). Nine of 42 (21.4%) patients with culture confirmed erythema migrans tested at the baseline visit, 42 of 118 (35.6%) adults, and 3 of 57 (5.3%) children whose sera were reactive for B. burgdorferi antibodies also tested positive for A. phagocytophila antibodies. The geometric mean titer ranged from 219 to 315 for all groups, and the differences in titers among the groups were not statistically significant. Only one-third of the healthy blood donors reactive by IFA were confirmed to be positive by immunoblotting. We conclude that a significant proportion of adults and children without clinical evidence of HGE will test positive for A. phagocytophila antibodies when the conventional cutoff titer of 80 is used in the IFA. This information must be considered in interpretation of test results. PMID- 12089288 TI - Improvement of AMPLICOR human immunodeficiency virus type 1 viral load test (version 1.5) by addition of a coprecipitant during the RNA isolation step. AB - The effect of the addition of a coprecipitant during the RNA isolation step on the analytical performance of the COBAS AMPLICOR human immunodeficiency virus type 1 (HIV-1) Monitor (version 1.5; Roche) viral load test was tested. Thirty six specimens including patient samples, positive control samples, and negative control samples were processed in the presence and absence of the Pellet Paint coprecipitant. Specimens processed without the coprecipitant had lower RNA yields, as evidenced by a lower signal for the quantitation standard (QS). In addition, the results for all samples processed with the coprecipitant were acceptable on the basis of the optical density (OD) reading for the QS, whereas the result for one specimen processed without the coprecipitant was unacceptable on the basis of the OD reading for the QS, which required the assay to be repeated. Furthermore, the use of the coprecipitant improved the overall precision of the assay. PMID- 12089289 TI - Use of the sensititre colorimetric microdilution panel for antifungal susceptibility testing of dermatophytes. AB - The Sensititre YeastOne antifungal panel was used to test 49 dermatophytes belonging to the species Epidermophyton floccosum, Microsporum gypseum, Microsporum canis, Trichophyton tonsurans, Trichophyton rubrum, and Trichophyton mentagrophytes. The MICs of four antifungals obtained with the Sensititre YeastOne antifungal panel were compared with those obtained by the reference NCCLS microdilution method. The levels of agreement between the two methods ( 0.05). Addition of bradykinin, enalaprilat, and amlodipine decreased oxygen consumption significantly less in SHR than WKY (SHR: bradykinin -13.9 +/- 1.9%, enalaprilat -15.3 +/- 1.6%, amlodipine -11.9 +/- 0.7%; WKY: bradykinin -22.8 +/- 1.0%, enalaprilat -24.1 +/- 2.0%, amlodipine -20.7 +/- 2.3%; P < 0.05), consistent with less NO effect in SHR. Addition of tempol reversed the defects in responsiveness to enalaprilat and amlodipine, suggesting that inactivation of NO by superoxide contributes to decreased NO availability. The response to an NO donor was similar in both groups and was unaffected by the addition of tempol. These results demonstrate that NO availability in the kidney is decreased in SHR, resulting in increased oxygen consumption. This effect is due to enhanced production of superoxide in SHR. By lowering intrarenal oxygen levels, reduced NO may contribute to susceptibility to injury and renal fibrosis. Increasing NO production, decreasing oxidant stress, or both might prevent these changes by improving renal oxygenation. PMID- 12089375 TI - Peritubular capillary regression during the progression of experimental obstructive nephropathy. AB - Injury to the renal microvasculature may be a major factor contributing to the progression of renal disease. Although severe disruption of peritubular capillaries (PTC) could lead to marked tubulointerstitial scarring, elucidation of that process remains incomplete. This study investigated the morphologic changes in PTC and their likely regulation by vascular endothelial growth factor (VEGF) during the progression of tubulointerstitial injuries. Unilateral ureteral obstruction was induced in Wistar rats by ligation of the left ureter, and the kidneys were then collected at selected times. PTC lumina and the expression of VEGF and its receptor Flk-1 were immunohistochemically detected. Morphologic changes in PTC endothelial cells were examined by using Ki67 staining, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling, and electron microscopic studies. In the first week of the disease period, immunohistochemical labeling of tubular VEGF intensified, with accompanying deformation and dilation of adjacent thrombomodulin (TM)-positive PTC lumina; an angiogenic response of endothelial cells was demonstrated with Ki67 and TM double-staining. During the subsequent 2 wk, tubular VEGF labeling decreased until it was virtually absent, an effect confirmed by Western blotting. Concomitantly, labeling of the VEGF receptor Flk-1 in PTC endothelial cells decreased and PTC lumina began to regress, demonstrating endothelial cell apoptosis (as detected in terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling and electron microscopic studies). By the end of week 4, the numbers of TM-positive PTC lumina were significantly decreased in areas of marked tubulointerstitial scarring. These results suggest that PTC regression, involving an early, unsustained, angiogenic response followed by progressive endothelial cell apoptosis, could be a potential factor contributing to tubulointerstitial scarring in this unilateral ureteral obstruction model. PMID- 12089376 TI - HIV-1 Nef induces proliferation and anchorage-independent growth in podocytes. AB - HIV-associated nephropathy (HIVAN) is now the third leading cause of end-stage renal disease in the African American population. HIV-1 infects renal tubular and glomerular epithelial cells or podocytes, cells that are a critical part of the filtration barrier. HIV-1 infection induces the loss of podocyte differentiation markers and increases podocyte proliferation. It has been previously shown that HIV-infection induces loss of contact inhibition. Here, the HIV-1 gene responsible for proliferative changes is identified by using cultured podocytes in vitro. The HIV-1 proviral construct, pNL4-3 was rendered noninfectious by replacing the HIV-1 gag/pol sequences with an EGFP reporter gene (pNL4-3: DeltaG/P-EGFP). This construct was then pseudotyped with VSV.G envelope to infect podocytes that were conditionally immortalized with SV-40 T antigen. In addition, mutated constructs were engineered with premature stop codons in the HIV-1 env, vif, vpr, vpu, nef, or rev genes. The parental construct and all the other mutated constructs, with the exception of nef, induced proliferation under nonpermissive conditions and anchorage-independent growth (colony formation in soft agar) under permissive conditions. In contrast, deletion of nef markedly reduced proliferation and colony formation. Although tat alone, or tat plus rev induced marginal levels of anchorage-independent growth, coexpression with nef significantly increased colony formation. Finally, stable expression of Nef in a retroviral vector, pBabe-puro, was sufficient to induce increased proliferation and colony formation. Moreover, nef induced saturation density and loss of contact inhibition. These data indicate that Nef induces multiple proliferative effects in podocytes in culture and that nef may therefore be an important gene in the pathogenesis of HIVAN in vivo. PMID- 12089377 TI - Mitochondrial DNA mutations in focal segmental glomerulosclerosis lesions. AB - Glomerular epithelial cells are primary pathogenic sites in focal segmental glomerulosclerosis (FGS) lesions. Glomerular epithelial cells are regarded as terminally differentiated cells that do not proliferate. These characteristics are also noted for neurons and muscular cells, which are major sites of mitochondrial DNA (mtDNA) mutation accumulation. Screening for mtDNA mutations was performed with renal biopsy specimens from patients with primary FGS and patients with IgA nephropathy (as subjects with secondary FGS and as control subjects). mtDNA extracted from kidney biopsy specimens was amplified with appropriate primer pairs for study of the mtDNA point mutations 3243A-->G, 3271T- >C, 8344A-->G, and 8993T-->G/C, as well as the common deletion (a 4977-bp deletion spanning mtDNA nucleotide pairs 8469 to 13447). In situ amplification of both total mtDNA and the common deletion was also performed. Two patients with FGS demonstrated the 3243A-->G point mutation; 12 patients with FGS and seven patients with IgA nephropathy accompanied by glomerulosclerotic lesions exhibited the common deletion in their kidney tissue. No patient demonstrated the mtDNA mutations 3271T-->C, 8344A-->G, or 8993T-->G/C. The degree of heteroplasmy for the 3243A-->G point mutation was >85%; however, the heteroplasmy for the common deletion was <1%. As determined with in situ PCR, normal mtDNA was mainly distributed in the tubular epithelium and mtDNA with the common deletion was mainly distributed among glomerular epithelial cells. In conclusion, it is suggested that mtDNA mutations are distributed in glomerular epithelial cells among some patients with primary FGS or secondary FGS with IgA nephropathy. These mutations may be related to glomerular epithelial cell damage. PMID- 12089378 TI - A minimal Ksp-cadherin promoter linked to a green fluorescent protein reporter gene exhibits tissue-specific expression in the developing kidney and genitourinary tract. AB - Ksp-cadherin is a unique, tissue-specific member of the cadherin family of cell adhesion molecules that is expressed exclusively in tubular epithelial cells in the kidney and developing genitourinary (GU) tract. Transgenic mice carrying 3425 bp of the Ksp-cadherin 5' flanking region linked to a lacZ reporter gene express beta-galactosidase exclusively in the kidney, although the expression pattern is incomplete (Am J Physiol 277: F599-F610, 1999). To further define the region that mediates tissue-specific expression, transgenic mice carrying 1341 bp or 324 bp of the 5' flanking region linked to a green fluorescent protein (GFP) reporter gene were produced. Transgenic mice carrying 1341 bp of the 5' flanking region expressed GFP in all embryonic tissues that endogenously express Ksp-cadherin, including the ureteric bud, Wolffian duct, Mullerian duct, and developing tubules in the mesonephros and metanephros. In the adult kidney, GFP was highly expressed in thick ascending limbs of Henle's loops and collecting ducts and weakly expressed in proximal tubules and Bowman's capsules. Transgenic mice carrying 324 bp of the 5' flanking region exhibited expression exclusively in tubular epithelial cells in the developing kidney and GU tract. Immunoblot analysis showed that the expression of GFP was restricted to the kidney in adult mice. Taken together, these results demonstrate that 324 bp of the Ksp-cadherin 5' flanking region is sufficient to direct epithelial-specific expression in the developing kidney and GU tract. Transgenic mice that express GFP in the mesonephros, metanephros, ureteric bud, and sex ducts may be useful for cell lineage studies. PMID- 12089379 TI - Epithelial-specific Cre/lox recombination in the developing kidney and genitourinary tract. AB - Ksp-cadherin is a unique, tissue-specific member of the cadherin family of cell adhesion molecules that is expressed in tubular epithelial cells in the kidney and developing genitourinary (GU) tract. It has recently been shown that a 1341 bp fragment of the 5' flanking region containing the Ksp-cadherin gene promoter can recapitulate the complete expression pattern of the gene in the developing kidney and GU tract. Similar to the endogenous Ksp-cadherin gene, transgenes containing 1341 bp of the 5' flanking region are expressed in developing nephrons, ureteric bud, mesonephric tubules, Wolffian duct, and Mullerian duct. In adult mice, the expression is restricted to renal tubules. In the current study, Ksp1.3/Cre transgenic mice carrying 1329 bp of the Ksp-cadherin 5' flanking region linked to the Cre recombinase gene were produced. Adult transgenic mice expressed Cre recombinase in renal tubules, especially collecting ducts and thick ascending limbs of Henle's loops. Transgenic embryos expressed Cre recombinase in the branching ureteric bud, developing renal tubules, and sex ducts. Ksp1.3/Cre transgenic mice were crossed with mice carrying a lacZ reporter gene that is activated by Cre/lox-mediated genetic recombination. Bitransgenic progeny expressed lacZ exclusively in renal tubules, mesonephric tubules, ureteric bud, developing ureter, and Wolffian duct. These results demonstrate that Ksp1.3/Cre transgenic mice express Cre recombinase exclusively in the kidney and developing GU tract and can mediate epithelial-specific Cre/lox recombination in these tissues. Ksp1.3/Cre transgenic mice should be useful for cell lineage studies and kidney-specific gene targeting. PMID- 12089380 TI - Association of the P-glycoprotein transporter MDR1(C3435T) polymorphism with the susceptibility to renal epithelial tumors. AB - Except for hereditary disease, genetic factors that contribute to the development of renal epithelial tumors are unknown. There is a possibility that the MDR1 encoded plasma membrane transporter P-glycoprotein (PGP) influences the risk of development of renal neoplasms. PGP is known to be involved in uptake, binding, transport, and distribution of xenobiotics. There is evidence that the MDR1(C3435T) polymorphism drives expression and modulates disease risk. In an explorational case-control study, constitutional genotype frequencies were established at MDR1(C3435T) of 537 healthy control subjects and compared with those of 212 patients with renal epithelial tumors. There were 179 clear cell renal cell carcinoma (CCRCC) and 33 tumors collectively assigned as non-CCRCC. In a second study, genotypes of another 150 healthy control subjects and 50 patients with three non-CCRCC types (26 papillary RCC, 11 chromophobe RCC, and 13 renal oncocytic adenoma) were compared. PCR-restriction fragment length polymorphism based analysis of constitutional DNA, and statistical analysis were applied. PGP expression was analyzed by quantitative immunohistochemistry. The explorational study showed a significant association between T allele frequency and the occurrence of tumors (P = 0.007). When tumors were histopathologically distinguished into frequent CCRCC and less frequent non-CCRCC, both patient groups contributed to this effect with a seemingly strong influence by the latter (P = 0.0419). The second study established the T allele as a risk factor especially for non-CCRCC (P = 0.0005) with the highest risk for homozygote TT allele carriers (P < 0.0001). Independently, MDR1(C3435T) genotype associated variations in PGP expression were shown in normal renal parenchyma with a 1.5 fold difference of median values (TT, 1.9; CC, 2.8; P = 0.0065). The data provide evidence for PGP to influence the susceptibility to develop renal epithelial tumors by virtue of its MDR1(C3435T) polymorphism and changes in expression. Especially T and TT carriers are at risk for developing non-CCRCC, i.e., papillary and chromophobe RCC as well as oncocytic adenomas. PMID- 12089381 TI - Altered expression pattern of polycystin-2 in acute and chronic renal tubular diseases. AB - Polycystin-2 represents one of so far two proteins found to be mutated in patients with autosomal-dominant polycystic kidney disease. Evidence obtained from experiments carried out in cell lines and with native kidney tissue strongly suggests that polycystin-2 is located in the endoplasmic reticulum. In the kidney, polycystin-2 is highly expressed in cells of the distal and connecting tubules, where it is located in the basal compartment. It is not known whether the expression of polycystin-2 in the kidney changes or whether it can be manipulated under certain instances. Therefore, the distribution of polycystin-2 under conditions leading to acute and chronic renal failure was analyzed. During ischemic acute renal failure, which affects primarily the S3 segment of the proximal tubule, a pronounced upregulation of polycystin-2 and a predominantly combined homogeneous and punctate cytoplasmic distribution in damaged cells was observed. After thallium-induced acute injury to thick ascending limb cells, polycystin-2 staining assumed a chicken wire-like pattern in damaged cells. In the (cy/+) rat, a model for autosomal-dominant polycystic kidney disease in which cysts originate predominantly from the proximal tubule, polycystin-2 immunoreactivity was lost in some distal tubules. In kidneys from (pcy/pcy) mice, a model for autosomal-recessive polycystic kidney disease in which cyst formation primarily affects distal tubules and collecting ducts, a minor portion of cyst lining cells cease to express polycystin-2, whereas in the remaining cells, polycystin-2 is retained in their basal compartment. Data show that the expression and cellular distribution of polycystin-2 in different kinds of renal injuries depends on the type of damage and on the nephron-specific response to the injury. After ischemia, polycystin-2 may be upregulated by the injured cells to protect themselves. It is unlikely that polycystin-2 plays a role in cyst formation in the (cy/+) rat and in the (pcy/pcy) mouse. PMID- 12089383 TI - Impaired renal sensory responses after renal ischemia in the rat. AB - Renal sensory responses and reflex function were examined in rats 24 h after 45 min of ischemic injury caused by unilateral renal arterial occlusion (RAO). The integrity of renal pelvic mechanoreceptor (MRu)-mediated renorenal reflex was examined. An increase in ipsilateral afferent renal nerve activity (ARNA) and a reflex decrease in efferent renal nerve activity (ERNA) and contralateral diuresis and natriuresis produced by increasing the intrapelvic pressure were seen in sham-operated (Sham) rats, but it was largely attenuated in RAO rats. Using single-fiber recordings of the renal MRu discharge, graded increases in intrapelvic pressure or renal pelvic administration of substance P (SP) resulted in pressure- or concentration-dependent increases in ARNA in the control kidney of Sham rats, whereas attenuated responses were seen in the postischemic kidney of RAO rats. The unresponsiveness of renal MRus in RAO rats was accompanied by an insufficient release of SP. However, the baseline SP release is higher in RAO kidneys due to a reduced neutral endopeptidase (NEP) activity in the renal pelvis of the postischemic kidney. No changes in NK-1 receptor mRNA levels were demonstrated; however, the expression of NK-1 receptors in the plasma membrane of RAO pelvis were decreased, possibly resulting from the internalization of the receptors associated with beta-arrestin trafficking. Renal excretory responses after saline loading were significantly lower in the postischemic kidney of RAO rats than in Sham rats. Responses of ARNA and ERNA were also lower. It is concluded that the defective activation of renal sensory mechanoreceptors in the postischemic kidney results from an inadequate release of SP after mechanostimulation and the reduced functional NK-1 receptors. PMID- 12089382 TI - Salt-sensitivity of proximal reabsorption alters macula densa salt and explains the paradoxical effect of dietary salt on glomerular filtration rate in diabetes mellitus. AB - GFR varies inversely with dietary NaCl in patients with early type I diabetes and in streptozotocin (STZ)-diabetic rats. To explain this paradox within the laws of physiology, it was hypothesized that it results from heightened sensitivity of the diabetic proximal tubule to dietary salt because changes in proximal reabsorption (Jprox) elicit reciprocal adjustments in GFR through the normal actions of tubuloglomerular feedback (TGF). Micropuncture was done in rats after 5 wk of moderately hyperglycemic STZ-diabetes and 1 wk of different NaCl diets. First, single-nephron GFR (SNGFR) and early distal tubular Na(+), Cl(-) and K(+) concentration (representing the TGF signal) were measured by collecting from early distal nephrons. In nondiabetics, dietary salt did not affect SNGFR or the TGF signal. In diabetics, the TGF signal varied directly with dietary salt while SNGFR varied inversely with dietary salt. Next, Jprox was measured by collecting from late proximal tubules. To control for different SNGFR, SNGFR was manipulated by perfusing Henle's loop to alter TGF activity. Controlling for SNGFR, dietary salt did not affect Jprox in nondiabetics but exerted a major inverse impact on Jprox in diabetics. In conclusion, normal rats acclimate to dietary NaCl by primarily adjusting transport downstream of the macula densa. In contrast, diabetes renders reabsorption in the proximal tubule sensitive to dietary NaCl with subsequent effects on the TGF signal. This explains the paradoxical effect of dietary NaCl on GFR in early diabetes. PMID- 12089384 TI - Estimating total body water in children on the basis of height and weight: a reevaluation of the formulas of Mellits and Cheek. AB - An estimate of total body water (TBW) has important implications in clinical practice. For patients on peritoneal dialysis (PD), the estimate is critical when determining the delivered dialysis dose. The formulas of Mellits and Cheek have been recommended to estimate TBW in children on PD. These formulas were derived from healthy children, and very few infants were included. To assess the accuracy of these formulas, the original data were obtained and additional data from a broad literature review were compiled. The majority of the new data points were in the infant age range. Data were fitted using least-squares methodology and backward elimination to obtain a parsimonious model. Best fits were obtained using age, gender, and weight or a height x weight term. The results of the curves are as follows: Infants 0 to 3 mo (n= 71): TBW = 0.887 x (Wt)(0.83) Children 3 mo to 13 yr(n = 167): TBW = 0.0846 x 0.95([if female]) x (Ht xWt)(0.65) Children > 13 yr (n = 99): TBW = 0.0758 x 0.84([if female]) x (Ht x Wt)(0.69) When compared with the previous Mellits and Cheek formulas, the new formula fits better for infants (comparison of prediction errors, P < 0.0004). These newer formulas do not perform significantly better for the older two groups. Actual TBW measurement in children on PD must still be determined to verify the use of these formulas and to accurately assess dialysis delivery and adequacy. PMID- 12089385 TI - Changing trends in the survival of dialysis patients with human immunodeficiency virus in the United States. AB - HIV-infected patients with end-stage renal disease have a very high morbidity and mortality. In the last decade, survival of HIV-infected patients in the United States has remarkably improved. To determine whether similar improvement in survival has occurred in HIV-infected dialysis patients, their survival was evaluated by using the United States Renal Data System database. Survival of HIV infected dialysis patients in the United States was determined and the influence of year of initiation of dialysis, and demographic characteristics on the survival were analyzed by the Kaplan-Meier method. The effects of above variables on survival were also examined in a Cox proportional hazards model. Identified were 6166 HIV-infected patients with end-stage renal disease who received dialysis in the United States. Eighty-nine percent of the patients were black, 7.4% white, and 3% other. From 1990 to 1999, 1-yr survival of HIV-infected patients on dialysis improved from 56 to 74%, and the annual death rates declined from 458 deaths to 240 deaths per 1000 patient-years. The hazard ratio declined significantly in patients who initiated dialysis in years 1999-2000 compared with patients who initiated dialysis < or = 1990 (hazard ratio, 0.49; 95% confidence interval, 0. 40 to 0.60). Survival of HIV-infected dialysis patients has remarkably improved in the United States. PMID- 12089386 TI - Insulin resistance as an independent predictor of cardiovascular mortality in patients with end-stage renal disease. AB - Insulin resistance is closely associated with atherosclerosis and cardiovascular mortality in the general population. Patients with end-stage renal disease (ESRD) are known to have insulin resistance, advanced atherosclerosis, and a high cardiovascular mortality rate. We evaluated whether insulin resistance is a predictor of cardiovascular death in a cohort of ESRD. A prospective observational cohort study was performed in 183 nondiabetic patients with ESRD treated with maintenance hemodialysis. Insulin resistance was evaluated by the homeostasis model assessment method (HOMA-IR) using fasting glucose and insulin levels at baseline, and the cohort was followed for a mean period of 67 mo. Forty nine deaths were recorded, including 22 cardiovascular deaths. Cumulative incidence of cardiovascular death by Kaplan-Meier estimation was significantly different between subjects in the top tertile of HOMA-IR (1.40 to 4.59) and those in the lower tertiles of HOMA-IR (0.28 to 1.39), and the hazard ratio (HR) was 2.60 (95% confidence interval [CI], 1.12 to 6.01; P = 0.026) in the univariate Cox proportional hazards model. In multivariate Cox models, the positive association between HOMA-IR and cardiovascular mortality remained significant (HR, 4.60; 95% CI, 1.83 to 11.55; P = 0.001) and independent of age, C-reactive protein, and presence of preexisting vascular complications. Further analyses showed that the effect of HOMA-IR on cardiovascular mortality was independent of body mass index, hypertension, and dyslipidemia. In contrast, HOMA-IR did not show such a significant association with noncardiovascular mortality. These results indicate that insulin resistance is an independent predictor of cardiovascular mortality in ESRD. PMID- 12089387 TI - Familial risk of preeclampsia in Newfoundland: a population-based study. AB - This study sought to quantify the familial risk of preeclampsia (proteinuric hypertension) in Newfoundland and to identify characteristics in probands that predict increased familial risk. Reviewed were 5173 obstetric charts from 10 hospitals, representing 99% of deliveries on the island of Newfoundland for a 1 yr period from April 1996 to March 1997; pregnancy-induced hypertension was diagnosed according to strict criteria. Family obstetric histories were obtained from identified probands with preeclampsia, and sisters and mothers of probands were interviewed. In addition, the obstetric charts from sisters and mothers were reviewed to identify preeclampsia. The incidence of preeclampsia in the population was 5.6% (n = 292), and in primiparous women it was 7.9%. Factors independently associated with increased risk of preeclampsia included primiparous delivery, multiple gestation, pregestational and gestational diabetes, maternal age of more than 35 yr, and region of the province. Of 330 sisters identified, 217 had 445 pregnancies, with 331 charts located for review. The incidence of preeclampsia (based on chart review) in 163 primiparous sisters was 20.2%. The relative risk of preeclampsia in primiparous sisters of probands with preeclampsia compared with primiparous women in the population was 2.6 (95% confidence interval, 1.8 to 3.6). Factors in probands independently associated with a higher risk of preeclampsia in sisters included at least 2+ proteinuria and region of the province. This population-based study, which used unbiased ascertainment and strict diagnostic criteria, demonstrated a significantly higher risk of preeclampsia in sisters of probands with preeclampsia, particularly when probands were defined by severity of preeclampsia and by geographic region. PMID- 12089388 TI - Risk factors for proteinuria in a large, multiracial, southeast Asian population. AB - The factors associated with proteinuria were examined in a large multiracial Asian population participating in a screening program aimed at the early detection of renal disease. Of 213,873 adults who participated, 189,117 with complete data were included. Malay race, increasing age, both extremes of body mass index (BMI), self-reported family history of kidney disease (FKD), and higher systolic and diastolic BP measurements (even at levels classified as being within the normal range) were independently associated with dipstick-positive proteinuria. The odds ratios (OR) for proteinuria increased progressively with age. There was a J-shaped relationship between BMI and proteinuria (OR of 1.3, 1.00, 1.3, 1.6, and 2.5 for BMI of < or =18.00, 23.00 to 24.99, 25.00 to 27.49, 27.50 to 29.99, and > or =30.00 kg/m(2), respectively, compared with BMI of 18.01 to 22.99 kg/m(2)). OR for proteinuria according to systolic and diastolic BP were significantly increased beginning at levels of 110 and 90 mmHg, respectively. In addition, the Malay race was associated with a significantly higher OR for proteinuria, compared with the Chinese race (OR of 1.3). Finally, FKD was significantly associated with proteinuria (OR of 1.7), whereas a family history of diabetes mellitus and a family history of hypertension were not. When family histories were analyzed by clustering, isolated FKD remained a significant determinant of proteinuria and the magnitude of the effect was not significantly different from that observed in the presence of a coexisting family history of diabetes mellitus or hypertension. This is the first study to evaluate factors associated with proteinuria in an Asian population. The epidemiologic study of renal disease in this population suggests that risk factors for renal disease might differ significantly among racial groups. PMID- 12089389 TI - Traditional cardiovascular disease risk factors in dialysis patients compared with the general population: the CHOICE Study. AB - Although atherosclerotic cardiovascular disease (ASCVD) risk in end-stage renal disease (ESRD) is 5 to 30 times that of the general population, few data exist comparing ASCVD risk factors among new dialysis patients to the general population. This cross-sectional study of 1041 dialysis patients describes the prevalence of ASCVD risk factors at the beginning of ESRD compared with estimates of ASCVD risk factors in the adult US population derived from the Third National Health and Nutrition Examination (NHANES III). CHOICE Study participants had a high prevalence of diabetes (54%), hypertension (96%), left ventricular hypertrophy by electrocardiogram (EKG) criteria (22%), low physical activity (80%), hypertriglyceridemia (36%), and low HDL cholesterol (33%). CHOICE participants were more likely to be older, black, and male than NHANES III participants. After adjustment for age, race, gender, and ASCVD (defined as myocardial infarction, revascularization procedure, stroke, carotid endarterectomy, and amputation in CHOICE; and as myocardial infarction and stroke in NHANES III), the prevalence of diabetes, hypertension, left ventricular hypertrophy by EKG, low physical activity, low HDL cholesterol, and hypertriglyceridemia were still more common in CHOICE participants. Smoking, obesity, hypercholesterolemia, and high LDL cholesterol, however, were less common in CHOICE than NHANES III participants. The projected 5-yr ASCVD risk based on the Framingham Risk Equation among those older than 40 yr without ASCVD was higher in CHOICE Study participants (13%) than in the NHANES III participants (6%). In summary, many ASCVD risk factors are more prevalent in ESRD than in the general population and may explain some, but probably not all, of the increased ASCVD risk in ESRD. PMID- 12089390 TI - Anemia and renal insufficiency are independent risk factors for death among patients with congestive heart failure admitted to community hospitals: a population-based study. AB - The purpose of this retrospective cohort study was to examine the associations among chronic kidney disease, anemia, and risk of death among patients with heart failure. Retrospective cohort study. Patients with a principal diagnosis of heart failure (ICD9 codes 402.01, 402.11, 402.91, 404.01, 404.11, 404.91, and 428.xx) were included. Chronic kidney disease (CKD) was defined as a serum creatinine >1.4 mg/dl for women and >1.5 mg/dl for men. There were 665 eligible patients in the sample with a mean (SD) age of 75.7 (10.9) yr; 60% were women, 71% were white, and 38% had CKD. On admission, a hematocrit > or =40% was found for 30.3% of the patients; 22.9% had a hematocrit between 36% and 40%, 33.2% between 30% and 35%, and 13.6% had a hematocrit of <30%. The 1-yr death rates among individuals with and without CKD were 44.9% and 31.4%, respectively (relative risk [RR], 1.43; 95% confidence interval [CI], 1.17 to 1.75). The mortality at 1 yr was 31.2% for individuals with a hematocrit > or =40%; 33.8% (RR, 1.08; 95% CI. 0.79 to 1.47) for hematocrit 36 to 39%; 36.7% (RR, 1.17; 95% CI, 0.89 to 1.54) for hematocrit between 30 and 35%; and 50.0% (RR, 1.60; 95% CI, 1.19 to 2.16) for those with a hematocrit <30% (chi(2) for trend was 7.37; P = 0.007). Both hematocrit and serum creatinine were independently associated with increased risk of death during follow-up after controlling for other patient risk factors. In conclusion, CKD and anemia are frequent among older patients with heart failure and are independent predictors of subsequent risk of death. PMID- 12089391 TI - Normalization of brain death-induced injury to rat renal allografts by recombinant soluble P-selectin glycoprotein ligand. AB - Donor brain death has been considered a significant risk factor for both early and late organ allograft dysfunction. This central injury not only evokes an upsurge of catecholamines with resultant peripheral tissue vasoconstriction and ischemia but also promotes release of hormones and inflammatory mediators that may also affect the organs directly. One of the resultant influences of these events is the rapid upregulation of the acute-phase adhesion molecules, the selectins. These initiate leukocyte adhesion to vascular endothelium and trigger subsequent cellular and molecular changes in the compromised tissues. An established F344 --> LEW rat model of chronic rejection was used to examine (1) whether the initial inflammatory events that develop within kidney allografts from brain-dead donors could be normalized using a recombinant soluble form of P selectin glycoprotein ligand and (2) whether amelioration of these early changes would alter the inexorable progression of chronic allograft rejection. Untreated living donor controls experienced unrelenting chronic rejection over time. This complex process was accelerated in brain-dead donor kidneys. Treatment with P selectin glycoprotein ligand prevented the early inflammatory changes in the transplanted organs and their subsequent (200 d) functional and morphologic manifestations, particularly when the soluble ligand was administered both to the donor before organ removal and to the recipient after engraftment. This strategy of using a naturally occurring selectin ligand to prevent donor-associated chronic graft dysfunction may be of special clinical interest in cadaver donor transplantation. PMID- 12089392 TI - Serum glomerular permeability activity in patients with podocin mutations (NPHS2) and steroid-resistant nephrotic syndrome. AB - A plasma factor displaying permeability activity in vitro and possibly determining proteinuria has been hypothesized in idiopathic focal segmental glomerulosclerosis (FSGS). In vitro permeability activity (P(alb)) was determined in sera of five patients with autosomal recessive steroid-resistant nephrotic syndrome (NPHS2), an inherited condition indistinguishable from idiopathic FSGS on clinical grounds, but in which proteinuria is determined by homozygous mutations of podocin, a key component of the glomerular podocyte. All patients had presented intractable proteinuria with nephrotic syndrome; four developed renal failure and received a renal allograft. For comparison, sera from 31 children with nephrotic syndrome were tested. Pretransplant P(alb) was high in all cases (mean 0.81 +/- 0.06), equivalent to levels observed in idiopathic FSGS. Overall, P(alb) did not correlate with proteinuria. The posttransplant outcome was complicated in two patients by recurrence of proteinuria after 10 and 300 d, respectively, that responded to plasmapheresis plus cyclophosphamide. P(alb) levels were high at the time of the recurrence episodes and steadily decreased after plasmapheresis, to reach normal levels in the absence of proteinuria after the seventh cycle. In an attempt to explain high P(alb) in these patients, putative inhibitors of the permeability activity were studied. Coincubation of serum with homologous nephrotic urine reduced P(alb) to 0, whereas normal urine did not determine any change, which suggests loss of inhibitory substances in nephrotic urine. The urinary levels of the serum P(alb) inhibitors apo J and apo E were negligible in all cases, thus suggesting that other urinary inhibitors were responsible for the neutralizing effect. These data indicate that P(alb) is high in NPHS2, probably resulting from loss of inhibitors in urine. Lack of correlation of P(alb) with proteinuria suggests a selective loss of inhibitors. As in idiopathic FSGS, proteinuria may also recur after renal transplantation in NPHS2 patients, and post-transplant proteinuria is associated with high P(alb). The relationship between elevated P(alb) and proteinuria in NPHS2 remains to be determined. PMID- 12089393 TI - ANCA-positive vasculitis. PMID- 12089394 TI - Repuncturing the renal biopsy: strategies for molecular diagnosis in nephrology. AB - As has been exemplified by recent progress in the classification of cancer, future approaches to enhance the clinical diagnostic power of tissue biopsies may be based on gene expression profiles. A series of strategies to translate these approaches to the diagnosis of renal disease is here proposed. The theoretical and technical problems resulting from the small amount of starting material available from renal biopsies will be specifically addressed. A preliminary study with cDNA array-based expression data obtained from kidneys with tubulointerstitial inflammation and fibrosis suggests the feasibility of distinguishing molecular categories of renal disease. Finally, a combined conventional and molecular work-up of renal biopsies will be suggested. These approaches should add a new dimension to biopsy interpretation and provide novel information concerning renal pathogenesis, diagnosis, prognosis, and differential therapy. A coordinated effort from nephrologists and pathologists in large multicenter trials will be required to achieve this goal. It is hoped that this outlook will lead to stimulating discussions and the implementation of these innovative ideas in nephrology. PMID- 12089396 TI - The breathing kidney. PMID- 12089397 TI - ANCA are pathogenic--oh yes they are! PMID- 12089410 TI - Steering a course to an AIDS vaccine. PMID- 12089411 TI - Intellectual property. Arrest of ex-Harvard postdocs raises questions of ownership. PMID- 12089412 TI - Paleontology. China regains fossils seized in California. PMID- 12089413 TI - Counterterrorism. Academies weigh in on homeland defense. PMID- 12089414 TI - Exoplanets. Winking star unveils planetary birthplace. PMID- 12089415 TI - Agbiotech. A little pollen goes a long way. PMID- 12089416 TI - Cancer research. Nanoparticles cut tumors' supply lines. PMID- 12089418 TI - Optics. New twist could pack photons with data. PMID- 12089417 TI - Patents. NIH to limit scope of foreign patents. PMID- 12089419 TI - High-energy physics. CERN panel calls for cuts and shake-ups. PMID- 12089420 TI - Astronomy. Cosmic lenses may be magnifying quasars. PMID- 12089421 TI - Canada. Few women win new academic chairs. PMID- 12089423 TI - Therapies. Raising the limits. PMID- 12089422 TI - Therapies. Confronting the limits of success. PMID- 12089424 TI - Therapies. The high cost of poverty. PMID- 12089425 TI - Vaccines. Monkey puzzles. PMID- 12089426 TI - Plant genetics. Something to sniff at: unbottling floral scent. PMID- 12089427 TI - Plant genetics. Plants 'speak' using versatile volatiles. PMID- 12089429 TI - Astronomy. White dwarf mergers and the rebirth of luminous stars. PMID- 12089428 TI - China and AIDS--the time to act is now. PMID- 12089430 TI - Signal transduction. MAP kinase signaling specificity. PMID- 12089431 TI - Molecular dynamics. Biomolecules see the light. PMID- 12089432 TI - Evolution. Chaperones as buffering agents? PMID- 12089433 TI - Supernova remnant OH masers: signposts of cosmic collision. AB - A supernova explosion, the final death throe of a massive star, creates an expanding bubble of hot gas that overruns up the surrounding medium. When a supernova remnant encounters a dense interstellar cloud, the compression may trigger gravitational collapse and the formation of a new generation of stars. This event can be detected through intense stimulated emission in the 1720 megahertz transition of the hydroxyl radical, OH, which yields unique insights into the physical processes and conditions occurring during the interaction. PMID- 12089435 TI - Separation of water into its ortho and para isomers. PMID- 12089434 TI - Diversity considerations in HIV-1 vaccine selection. AB - Globally, human immunodeficiency virus-type 1 (HIV-1) is extraordinarily variable, and this diversity poses a major obstacle to AIDS vaccine development. Currently, candidate vaccines are derived from isolates, with the hope that they will be sufficiently cross-reactive to protect against circulating viruses. This may be overly optimistic, however, given that HIV-1 envelope proteins can differ in more than 30% of their amino acids. To contend with the diversity, country specific vaccines are being considered, but evolutionary relationships may be more useful than regional considerations. Consensus or ancestor sequences could be used in vaccine design to minimize the genetic differences between vaccine strains and contemporary isolates, effectively reducing the extent of diversity by half. PMID- 12089436 TI - Strongly correlated superconductivity. AB - High-temperature superconductivity in doped Mott insulators such as the cuprates contradicts the conventional wisdom that electron repulsion is detrimental to superconductivity. Because doped fullerene conductors are also strongly correlated, the recent discovery of high-critical-temperature, presumably s-wave, superconductivity in C60 field effect devices is even more puzzling. We examine a dynamical mean-field solution of a model for electron-doped fullerenes that shows how strong correlations can indeed enhance superconductivity close to the Mott transition. We argue that the mechanism responsible for this enhancement could be common to a wider class of strongly correlated models, including those for cuprate superconductors. PMID- 12089437 TI - Nitrate controls on iron and arsenic in an urban lake. AB - Aquatic ecosystems are often contaminated by multiple substances. Nitrate, a common aquatic pollutant, strongly influenced the cycling of arsenic (As) under anoxic conditions in urban Upper Mystic Lake (Massachusetts, USA) by oxidizing ferrous iron [Fe(II)] to produce As-sorbing particulate hydrous ferric oxides and causing the more oxidized As(V), which is more particle-reactive than As(III) under these conditions, to dominate. This process is likely to be important in many natural waters. PMID- 12089438 TI - 50 million years of genomic stasis in endosymbiotic bacteria. AB - Comparison of two fully sequenced genomes of Buchnera aphidicola, the obligate endosymbionts of aphids, reveals the most extreme genome stability to date: no chromosome rearrangements or gene acquisitions have occurred in the past 50 to 70 million years, despite substantial sequence evolution and the inactivation and loss of individual genes. In contrast, the genomes of their closest free-living relatives, Escherichia coli and Salmonella spp., are more than 2000-fold more labile in content and gene order. The genomic stasis of B. aphidicola, likely attributable to the loss of phages, repeated sequences, and recA, indicates that B. aphidicola is no longer a source of ecological innovation for its hosts. PMID- 12089439 TI - A tropical rainforest in Colorado 1.4 million years after the Cretaceous-Tertiary boundary. AB - An extremely diverse lower Paleocene (64.1 million years ago) fossil leaf site from Castle Rock, Colorado, contains fossil litter that is similar to the litter of extant equatorial rainforests. The presence of a high-diversity tropical rainforest is unexpected, because other Paleocene floras are species-poor, a feature generally attributed to the Cretaceous-Tertiary (K-T) extinction. The site occurs on the margin of the Denver Basin in synorogenic sedimentary rocks associated with the rise of the Laramide Front Range. Orographic conditions caused by local topography, combined with equable climate, appear to have allowed for the establishment of rainforests within 1.4 million years of the K-T boundary. PMID- 12089440 TI - Proterozoic modular biomineralized metazoan from the Nama Group, Namibia. AB - We describe a Proterozoic, fully biomineralized metazoan from the Omkyk Member (approximately 549 million years before the present) of the northern Nama Group, Namibia. Namapoikia rietoogensis gen. et sp. nov. is up to 1 meter in diameter and bears a complex and robust biomineralized skeleton; it probably represents a cnidarian or poriferan. Namapoikia encrusts perpendicular to the walls of vertical synsedimentary fissures in microbial reefs. This finding implies that large, modular metazoans with biologically controlled mineralization appeared some 15 million years earlier than previously documented. PMID- 12089441 TI - Pollen-mediated movement of herbicide resistance between commercial canola fields. AB - There is considerable public and scientific debate for and against genetically modified (GM) crops. One of the first GM crops, Brassica napus (oilseed rape or canola) is now widely grown in North America, with proposed commercial release into Australia and Europe. Among concerns of opponents to these crops are claims that pollen movement will cause unacceptable levels of gene flow from GM to non GM crops or to related weedy species, resulting in genetic pollution of the environment. Therefore, quantifying pollen-mediated gene flow is vital for assessing the environmental impact of GM crops. This study quantifies at a landscape level the gene flow that occurs from herbicide-resistant canola crops to nearby crops not containing herbicide resistance genes. PMID- 12089442 TI - Myeloperoxidase, a leukocyte-derived vascular NO oxidase. AB - Myeloperoxidase (MPO) is an abundant mammalian phagocyte hemoprotein thought to primarily mediate host defense reactions. Although its microbicidal functions are well established in vitro, humans deficient in MPO are not at unusual risk of infection. MPO was observed herein to modulate the vascular signaling and vasodilatory functions of nitric oxide (NO) during acute inflammation. After leukocyte degranulation, MPO localized in and around vascular endothelial cells in a rodent model of acute endotoxemia and impaired endothelium-dependent relaxant responses, to which MPO-deficient mice were resistant. Altered vascular responsiveness was due to catalytic consumption of NO by substrate radicals generated by MPO. Thus MPO can directly modulate vascular inflammatory responses by regulating NO bioavailability. PMID- 12089443 TI - Channelrhodopsin-1: a light-gated proton channel in green algae. AB - Phototaxis and photophobic responses of green algae are mediated by rhodopsins with microbial-type chromophores. We report a complementary DNA sequence in the green alga Chlamydomonas reinhardtii that encodes a microbial opsin-related protein, which we term Channelopsin-1. The hydrophobic core region of the protein shows homology to the light-activated proton pump bacteriorhodopsin. Expression of Channelopsin-1, or only the hydrophobic core, in Xenopus laevis oocytes in the presence of all-trans retinal produces a light-gated conductance that shows characteristics of a channel selectively permeable for protons. We suggest that Channelrhodopsins are involved in phototaxis of green algae. PMID- 12089444 TI - A low genomic number of recessive lethals in natural populations of bluefin killifish and zebrafish. AB - Despite the importance of selection against deleterious mutations in natural populations, reliable estimates of the genomic numbers of mutant alleles in wild populations are scarce. We found that, in wild-caught bluefin killifish Lucania goodei (Fundulidae) and wild-caught zebrafish Danio rerio (Cyprinidae), the average numbers of recessive lethal alleles per individual are 1.9 (95% confidence limits 1.3 to 2.6) and 1.4 (95% confidence limits 1.0 to 2.0), respectively. These results, together with data on several Drosophila species and on Xenopus laevis, show that phylogenetically distant animals with different genome sizes and numbers of genes carry similar numbers of lethal mutations. PMID- 12089445 TI - AGTR2 mutations in X-linked mental retardation. AB - Two angiotensin II (Ang II)-specific receptors, AGTR1 and AGTR2, are expressed in the mammalian brain. Ang II actions on blood pressure regulation, water electrolyte balance, and hormone secretion are primarily mediated by AGTR1. The function of AGTR2 remains unclear. Here, we show that expression of the AGTR2 gene was absent in a female patient with mental retardation (MR) who had a balanced X;7 chromosomal translocation. Additionally, 8 of 590 unrelated male patients with MR were found to have sequence changes in the AGTR2 gene, including one frameshift and three missense mutations. These findings indicate a role for AGTR2 in brain development and cognitive function. PMID- 12089446 TI - Tumor regression by targeted gene delivery to the neovasculature. AB - Efforts to influence the biology of blood vessels by gene delivery have been hampered by a lack of targeting vectors specific for endothelial cells in diseased tissues. Here we show that a cationic nanoparticle (NP) coupled to an integrin alphavbeta3-targeting ligand can deliver genes selectively to angiogenic blood vessels in tumor-bearing mice. The therapeutic efficacy of this approach was tested by generating NPs conjugated to a mutant Raf gene, ATPmu-Raf, which blocks endothelial signaling and angiogenesis in response to multiple growth factors. Systemic injection of the NP into mice resulted in apoptosis of the tumor-associated endothelium, ultimately leading to tumor cell apoptosis and sustained regression of established primary and metastatic tumors. PMID- 12089447 TI - Ongoing modification of Mediterranean Pleistocene sapropels mediated by prokaryotes. AB - Late Pleistocene organic-rich sediments (sapropels) from the eastern Mediterranean Sea harbor unknown, metabolically active chemoorganotrophic prokaryotes. As compared to the carbon-lean intermediate layers, sapropels exhibit elevated cell numbers, increased activities of hydrolytic exoenzymes, and increased anaerobic glucose degradation rates, suggesting that microbial carbon substrates originate from sapropel layers up to 217,000 years old. 16S ribosomal RNA gene analyses revealed that as-yet-uncultured green nonsulfur bacteria constitute up to 70% of the total microbial biomass. Crenarchaeota constitute a smaller fraction (on average, 16%). A slow but significant turnover of glucose could be detected. Apparently, sapropels are still altered by the metabolic activity of green nonsulfur bacteria and crenarchaeota. PMID- 12089448 TI - Correction of ADA-SCID by stem cell gene therapy combined with nonmyeloablative conditioning. AB - Hematopoietic stem cell (HSC) gene therapy for adenosine deaminase (ADA) deficient severe combined immunodeficiency (SCID) has shown limited clinical efficacy because of the small proportion of engrafted genetically corrected HSCs. We describe an improved protocol for gene transfer into HSCs associated with nonmyeloablative conditioning. This protocol was used in two patients for whom enzyme replacement therapy was not available, which allowed the effect of gene therapy alone to be evaluated. Sustained engraftment of engineered HSCs with differentiation into multiple lineages resulted in increased lymphocyte counts, improved immune functions (including antigen-specific responses), and lower toxic metabolites. Both patients are currently at home and clinically well, with normal growth and development. These results indicate the safety and efficacy of HSC gene therapy combined with nonmyeloablative conditioning for the treatment of SCID. PMID- 12089449 TI - Systematic identification of pathways that couple cell growth and division in yeast. AB - Size homeostasis in budding yeast requires that cells grow to a critical size before commitment to division in the late prereplicative growth phase of the cell cycle, an event termed Start. We determined cell size distributions for the complete set of approximately 6000 Saccharomyces cerevisiae gene deletion strains and identified approximately 500 abnormally small (whi) or large (lge) mutants. Genetic analysis revealed a complex network of newly found factors that govern critical cell size at Start, the most potent of which were Sfp1, Sch9, Cdh1, Prs3, and Whi5. Ribosome biogenesis is intimately linked to cell size through Sfp1, a transcription factor that controls the expression of at least 60 genes implicated in ribosome assembly. Cell growth and division appear to be coupled by multiple conserved mechanisms. PMID- 12089450 TI - Myelin-associated glycoprotein as a functional ligand for the Nogo-66 receptor. AB - Axonal regeneration in the adult central nervous system (CNS) is limited by two proteins in myelin, Nogo and myelin-associated glycoprotein (MAG). The receptor for Nogo (NgR) has been identified as an axonal glycosyl-phosphatidyl-inositol (GPI)-anchored protein, whereas the MAG receptor has remained elusive. Here, we show that MAG binds directly, with high affinity, to NgR. Cleavage of GPI-linked proteins from axons protects growth cones from MAG-induced collapse, and dominant negative NgR eliminates MAG inhibition of neurite outgrowth. MAG-resistant embryonic neurons are rendered MAG-sensitive by expression of NgR. MAG and Nogo 66 activate NgR independently and serve as redundant NgR ligands that may limit axonal regeneration after CNS injury. PMID- 12089451 TI - Analytic and algorithmic solution of random satisfiability problems. AB - We study the satisfiability of random Boolean expressions built from many clauses with K variables per clause (K-satisfiability). Expressions with a ratio alpha of clauses to variables less than a threshold alphac are almost always satisfiable, whereas those with a ratio above this threshold are almost always unsatisfiable. We show the existence of an intermediate phase below alphac, where the proliferation of metastable states is responsible for the onset of complexity in search algorithms. We introduce a class of optimization algorithms that can deal with these metastable states; one such algorithm has been tested successfully on the largest existing benchmark of K-satisfiability. PMID- 12089453 TI - Structure of nanocrystalline anatase solved and refined from electron powder data. AB - Energy-filtered Debye-Scherrer electron powder data have been successfully employed to determine the structure of nanocrystalline anatase (TiO2). The performed structure analysis includes determining the unit cell, space group, solving the structure via direct methods from extracted intensities and refining the structure using the Rietveld technique. The refined structural parameters for space group I4(1)/amd are a = 3.872 (2), c = 9.616 (5) A with titanium at 0.5,0.75,0.375 and oxygen at 0.5,0.75,0.1618 (6). The obtained structure indicates low internal stress as judged from the almost regular geometry of the TiO6 building blocks. Striking resemblance with the anatase structure determined previously by Burdett, Hughbanks, Miller, Richardson & Smith [J. Am. Chem. Soc. (1987). 109, 3639-3646] from neutron diffraction on coarse-grained material gives strong support for the correctness of the structure determined here. The result of the present study shows that the methods originally developed for determining structures from X-ray powder data work equally well with data from electron powder diffraction. This may open the window for structural investigations on the vast number of nanocrystalline materials and thin films whose structures are difficult to determine by X-ray diffraction since they are frequently only available in small quantities. PMID- 12089454 TI - A maximum-likelihood method for global-optimization-based structure determination from powder diffraction data. AB - A maximum-likelihood algorithm has been incorporated into a crystal structure determination from a powder diffraction data framework that uses an integrated intensity-based global optimization technique. The algorithm is appropriate when the structural model being optimized is not a complete description of the crystal structure under study. PMID- 12089455 TI - Group-theoretical and geometrical considerations of the phase transition between the high-temperature polymorphs of quartz and tridymite. AB - A model was derived for the temperature-dependent phase transition between the high-temperature polymorphs of quartz (P6(4)22) and tridymite (P6(3)/mmc). Only the Si framework is considered, and the transformation can be described as a deformation of a homogeneous sphere packing with three contacts per sphere (type 3/10/h1) in the common subgroup P6(1)22 of P6(4)22 and P6(3)/mmc. The proposed model guarantees the three-dimensional connection of the crystal structure during the whole transformation process. PMID- 12089456 TI - Morphological possibilities in general crystallography. Snow crystals. AB - Morphological features of snow crystals are analyzed on the basis of concepts of a general crystallography, where point groups of infinite order are possible. The observations are first formulated in a set of rules, leading to a macroscopic growth lattice and to continuous growth boundaries. Both are brought in connection with two-dimensional integral invertible transformations. Families of boundaries are considered, labeled by a set of indices restricted by selection rules and generalizing the law of rational indices. These properties are indicated graphically on a sample of 12 natural snow crystals. Their geometric and arithmetic properties are summarized in a table. PMID- 12089457 TI - Structure, odd lines and topological entropy of disorder of amorphous silicon. AB - A continuous random network model of amorphous silicon, subject to periodic boundary conditions, is partitioned into cells bounded by irreducible rings. An algorithm has been developed to find the cells and the rings that bound them. A thread can be imagined to pass through odd rings (rings containing an odd number of atoms) without passing through even rings. Such a thread is an algorithmic realization of an odd line, which is the only topological defect in glass or amorphous condensed matter. The topological entropy of disorder associated with these odd lines is found to be approximately 80% of the value for an ideal tetrahedrally bonded random network of atoms for which the rings that bound the cells are statistically independent. PMID- 12089458 TI - A multigrid approach to the average lattices of quasicrystals. AB - An average structure associated with a given quasilattice is a system composed of several average lattices that in reciprocal space produces strong main reflections. The average lattice of a quasicrystal is a useful concept closely related to the geometric description of the quasicrystal to crystal transformation and has been proved to be structurally significant. Here we calculate average structures for arbitrary two- and three-dimensional quasilattices using the dual generalized method. Additionally, closed analytical expressions for the coordinates of the average structure, the quasiperiodic lattice and its diffraction pattern are given. PMID- 12089459 TI - Algebraic direct methods for few-atoms structure models. AB - As a basis for direct-methods phasing at very low resolution for macromolecular crystal structures, normalized structure-factor algebra is presented for few atoms structure models with N = 1, 2, 3, em leader equal atoms or polyatomic globs per unit cell. Main results include: [see text]. Triplet discriminant Delta(hk) and triplet weight W(hk) parameters, a approximately 4.0 and b approximately 3.0, respectively, were determined empirically in numerical error analyses. Tests with phases calculated for few-atoms 'super-glob' models of the protein apo-D-glyceraldehyde-3-phosphate dehydrogenase (approximately 10000 non-H atoms) showed that low-resolution phases from the new few-atoms tangent formula were much better than conventional tangent formula phases for N = 2 and 3; phases from the two formulae were essentially the same for N > or = 4. PMID- 12089460 TI - A new method for calculation of crystal susceptibilities for X-ray diffraction at arbitrary wavelength. AB - A novel method for the calculation of the X-ray susceptibility of a crystal in a wide range of radiation wavelengths is described. An analytical interpolation of one-electron wave functions is built to approximate the solution to Hartree-Fock equations for all atoms and ions of the periodic system of elements with high accuracy. These functions allow the calculation of the atomic form factors in the entire range of a transmitted momentum as well as the description of their anisotropy taking into account external and intracrystalline fields. Also, an analytical approximation for the force matrix of an arbitrary crystal is obtained and the microscopic calculation of the Debye-Waller factor for crystals with a complicated unit cell is presented. PMID- 12089461 TI - Absolute structure-factor measurements of an Al-Pd-Mn quasicrystal. AB - A number of X-ray reflections from an icosahedral quasicrystal Al-Pd-Mn have been measured with great accuracy on an absolute basis by making use of Bragg-case diffraction. Since the specimen had high crystal quality, the dynamical theory was used for analyzing the results and to extract structure factors from measured integrated intensities. Good agreement was found between theory and experiment for strong reflections. Anomalous transmission was found to be strong in the 'good' regions of the quasicrystalline specimen and it was measured on an absolute basis, but the small residual strains present in the specimen prevented an accurate comparison between theory and experiment. A detailed discussion is presented on the parameters that mostly affect anomalous transmission in relationship to the adopted structural model. PMID- 12089462 TI - Atomic structure of the (Al,Si)CuFe cubic approximant phase. AB - The structure of the alpha-(Al,Si)CuFe approximant phase is determined by a single-crystal X-ray diffraction study and compared to the ideal structure obtained by the perpendicular shear method of the parent icosahedral phase. It is shown that the local environments (typical atomic clusters) of the two phases are similar and expand significantly farther than the size of the unit cell of the approximant. The orbit Al(2) issuing from the theoretical icosahedral model corresponding to the inner dodecahedron of the Mackay-type cluster is not found in the approximant and is replaced by a partially occupied inner icosahedron with an unusually large Debye-Waller factor. PMID- 12089464 TI - The Goos-Hanchen effect at Bragg diffraction. AB - The strong incident-angle dependence of the phase of complex reflectivity causes a shift of the reflected beam from the geometrically expected path. This effect, known as the Goos-Hanchen effect in the visible region, was observed for Bragg case diffraction in the hard X-ray region. The shift was found to be in good agreement with the theory. PMID- 12089463 TI - On the symmetry of 9- and 10-hedra. AB - The symmetry point groups for all combinatorially non-isomorphic 9- and 10-hedra (2606 and 32300, respectively) are contributed in the paper for the first time. The most symmetrical polyhedra of 3 to 32 automorphism group orders (50 and 187, respectively) are drawn in the Schlegel projections and characterized by the facet symbols and symmetry point groups. PMID- 12089465 TI - Form factors for core electrons useful for the application of quantum crystallography (QCr) to organic molecules. AB - Form factors are calculated for the core electrons of the first-row atoms B, C, N, O and F. The form factors are presented in an analytical form, as appears in International Tables for X-ray Crystallography [Ibers & Hamilton (1974), Vol. IV, pp. 103-145. Birmingham: Kynoch Press; present distributor Kluwer Academic Publishers, Dordrecht]. Having such form factors for the core electrons reduces the number of parameters to be determined by the methods of quantum crystallography (QCr). It is shown that the form factors obtained are quite accurate. Thus, when they are used in QCr, they are expected to be consistent with accurate charge densities. PMID- 12089466 TI - Amphibians as environmental sentinels. PMID- 12089467 TI - Dichotomies in the perception of speech. PMID- 12089468 TI - Analysing phenotypic variation: when old-fashioned means up-to-date. PMID- 12089469 TI - Theoretical approaches to holistic biological features: Pattern formation, neural networks and the brain-mind relation. AB - The topic of this article is the relation between bottom-up and top-down, reductionist and holistic approaches to the solution of basic biological problems. While there is no doubt that the laws of physics apply to all events in space and time, including the domains of life, understanding biology depends not only on elucidating the role of the molecules involved, but, to an increasing extent, on systems theoretical approaches in diverse fields of the life sciences. Examples discussed in this article are the generation of spatial patterns in development by the interplay of autocatalysis and lateral inhibition; the evolution of integrating capabilities of the human brain, such as cognition-based empathy; and both neurobiological and epistemological aspects of scientific theories of consciousness and the mind. PMID- 12089470 TI - Carbon - the first frontier of information processing. AB - Information is often encoded as an aperiodic chain of building blocks. Modern digital computers use bits as the building blocks, but in general the choice of building blocks depends on the nature of the information to be encoded. What are the optimal building blocks to encode structural information? This can be analysed by substituting the operations of addition and multiplication of conventional arithmetic with translation and rotation. It is argued that at the molecular level, the best component for encoding discretized structural information is carbon. Living organisms discovered this billions of years ago, and used carbon as the back-bone for constructing proteins that function according to their structure. Structural analysis of polypeptide chains shows that an efficient and versatile structural language of 20 building blocks is needed to implement all the tasks carried out by proteins. Properties of amino acids indicate that the present triplet genetic code was preceded by a more primitive one, coding for 10 amino acids using two nucleotide bases. PMID- 12089471 TI - An insight into the possible mechanism of working of two-cistronic gene expression systems and rational designing of newer systems. AB - The initial attempts at hyper-expressing buffalo/goat growth hormone (GH)-ORFs in Escherichia coli directly under various strong promoters were not successful despite the presence of a functional gene. High level expression of GH was achieved as a fusion protein with glutathione-S-transferase (GST). To produce native GH in an unfused state, we adapted an established strategy of two cistronic approach in our system. In this strategy, utilizing one of the highly efficient reported sequences as the first cistron led to a nearly 1000-fold enhancement in the level of expression under an E. coli promoter (trc). In search of a newer first-cistron sequence as well as to see the generality of the two cistronic approach, we explored the ability of different lengths of a highly expressing natural gene to act as an efficient first cistron. Surprisingly, GST, which is naturally highly expressible in E. coli, could not be fitted into a successful two-cistronic construct. In addition, placement of the entire two cistronic expression cassette (which had earlier given high-level GH expression under trc promoter) under the T7 promoter in E. coli failed to hyper-express GH. These results suggest that the successful exploitation of the two-cistron arrangement for hyper-expression of eukaryotic ORFs in bacteria is not as straightforward as was previously thought. It appears probable that factors such as the sequence context, together with the length and codons used in the first cistron are important as well. PMID- 12089472 TI - Overexpression and characterization of dimeric and tetrameric forms of recombinant serine hydroxymethyltransferase from Bacillus stearothermophilus. AB - Serine hydroxymethyltransferase (SHMT), a pyridoxal-5' -phosphate (PLP) dependent enzyme catalyzes the interconversion of L-Ser and Gly using tetrahydrofolate as a substrate. The gene encoding for SHMT was amplified by PCR from genomic DNA of Bacillus stearothermophilus and the PCR product was cloned and overexpressed in Escherichia coli. The purified recombinant enzyme was isolated as a mixture of dimer (90%) and tetramer (10%). This is the first report demonstrating the existence of SHMT as a dimer and tetramer in the same organism. The specific activities at 37 C of the dimeric and tetrameric forms were 6 7 U/mg and 4 1 U/mg, respectively. The purified dimer was extremely thermostable with a T(m) of 85 degrees C in the presence of PLP and L-Ser. The temperature optimum of the dimer was 80 degrees C with a specific activity of 32 4 U/mg at this temperature. The enzyme catalyzed tetrahydrofolate-independent reactions at a slower rate compared to the tetrahydrofolate-dependent retro-aldol cleavage of L-Ser. The interaction with substrates and their analogues indicated that the orientation of PLP ring of B. stearothermophilus SHMT was probably different from sheep liver cytosolic recombinant SHMT (scSHMT). PMID- 12089473 TI - Human placental lipid induces mitogenesis and melanogenesis in B16F10 melanoma cells. AB - A hydroalcoholic extract of fresh term human placenta was found to be mitogenic as well as melanogenic on B16F10 mouse melanoma in an in vitro culture. The extract, a reservoir of a large number of bioactive molecules, was resolved to get the lipid fraction. Its activity was evaluated on B16F10 mouse melanoma by assessing the change in cellular morphology, growth and melanin induction. The lipid fraction, placental total lipid fraction (PTLF) tested in the study employed doses of 0 01 to 200 microg/ml; optimum growth and melanization accompanied by morphological changes were recorded at 10 and 100 microg/ml respectively. At intermediate doses growth and melanization were found to show a pattern of change over between growth and melanization and finally reached at an inverse relation at the respective optimal dose of response. Compared with defined sphingolipids, C(2) ceramide and sphingosine-1-phosphate, the results were mostly corroborative. The duality of biological response of sphingolipids as reported in numerous studies was comparable for the PTLF suggesting that its active component is a sphingolipid and showing its use for pigment recovery in vitiligo. PMID- 12089474 TI - Cloning and sequencing of complete tau-crystallin cDNA from embryonic lens of Crocodylus palustris. AB - tau-Crystallin is a taxon-specific structural protein found in eye lenses. We present here the cloning and sequencing of complete tau-crystallin cDNA from the embryonic lens of Crocodylus palustris and establish it to be identical to the a enolase gene from non-lenticular tissues. Quantitatively, the tau-crystallin was found to be the least abundant crystallin of the crocodilian embryonic lenses. Crocodile tau-crystallin cDNA was isolated by RT-PCR using primers designed from the only other reported sequence from duck and completed by 5'- and 3'-rapid amplification of cDNA ends (RACE) using crocodile gene specific primers designed in the study. The complete tau-crystallin cDNA of crocodile comprises 1305 bp long ORF and 92 and 409 bp long untranslated 5'- and 3'-ends respectively. Further, it was found to be identical to its putative counterpart enzyme a enolase, from brain, heart and gonad, suggesting both to be the product of the same gene. The study thus provides the first report on cDNA sequence of tau crystallin from a reptilian species and also re-confirms it to be an example of the phenomenon of gene sharing as was demonstrated earlier in the case of peking duck. Moreover, the gene lineage reconstruction analysis helps our understanding of the evolution of crocodilians and avian species. PMID- 12089476 TI - Water temperature and pH influence olfactory sensitivity to pre-ovulatory and post-ovulatory ovarian pheromones in male Barilius bendelisis. AB - The attractive response and sexual activity elicited by pre-ovulatory steroid sulphate and post-ovulatory 15K-PGF pheromones are greater in wild caught tubercular males and immature males which express breeding tubercles on the snout (at 12-13 days post androgen implant) than in non-tubercular and non-androgen implanted males of freshwater fish Barilius bendelisis. This shows that circulatory androgens exert an activational effect on olfactory receptors of male fish. Wild caught tubercular males and androgen implanted juvenile males exhibit a high responsiveness to steroid sulphate at the water temperature and pH which fish experience during the pre-spawning phase. The male's sensitivity to 15K-PGF is almost equally high at the water temperature and pH which they experience in wild during the both pre-spawning and spawning periods. This suggests that the differential olfactory sensitivity to the two classes of pheromones in androgen implanted males is due to the varied temperature and pH of water, and that during the breeding season the male's olfactory sensitivity to PGF pheromone is more widespread than to the steroidal pheromone. An increased and decreased olfactory sensitivity in mature males to sex pheromones and L-alanine respectively during the breeding phase is in agreement with the hypothesis that pheromonal stimuli dominate over feeding stimuli to promote spawning success. PMID- 12089478 TI - A theropod tooth from the Late Triassic of southern Africa. AB - An isolated, large recurved and finely serrated tooth found associated with the prosauropod Euskelosaurus fron the Late Triassic part of the Elliot Formation is described here. It is compared to the Triassic thecodonts and carnivorous dinosaurs and its possible affinity is discussed. The tooth possibly belongs to a basal theropod and shows some features similar to the allosauroids. This tooth is of significance, as dinosaur remains except for some footprints and trackways, are poorly known in the Late Triassic horizons of southern Africa. PMID- 12089477 TI - Cocoon production, morphology, hatching pattern and fecundity in seven tropical earthworm species - a laboratory-based investigation. AB - Data on the reproductive biology of seven Indian species of earthworms, viz. Perionyx excavatus Perrier, Lampito mauritii Kinberg, Polypheretima elongata (Perrier), Pontoscolex corethrurus (Muller), Eutyphoeus gammiei (Beddard), Dichogaster modiglianii (Rosa) and Drawida nepalensis Michaelsen are presented. The peregrine earthworms such as Perionyx excavatus, Pontoscolex corethrurus, Dichogaster modiglianii, and Polypheretima elongata are considered to be continuous breeders with high fecundity. Native Lampito mauritii and Drawida nepalensis are semi-continuous and Eutyphoeus gammiei discrete breeders. There is a dramatic increase in cocoon production by most earthworm species of Tripura in the summer and monsoon with a corresponding peak during April and July. Cocoon production decreased or ceased during winter. Temperature affected the incubation period of cocoons. With increase in temperature, incubation period increased in the endogeic worms, Pontoscolex corethrurus, Polypheretima elongata and Drawida nepalensis and decreased in the epigeic worms, Perionyx excavatus and Dichogaster modiglianii, within a temperature range between 28-32 degrees C under laboratory conditions. There was a significant (P < 0.05) positive correlation between number of hatchlings per cocoon and incubation period in Lampito mauritii. High rate of cocoon production, short development time with high hatching success, as well as continuous breeding strategies in the epigeic species Perionyx excavatus and Dichogaster modiglianii and the top soil endogeic species, Pontoscolex corethrurus, Drawida nepalensis and Lampito mauritii, indicate their possible usefulness in vermiculture. The giant anecic worm, Eutyphoeus gammiei, which has a very long cocoon development time, discrete breeding strategy and very low rate of cocoon production, is not a suitable species for vermiculture. PMID- 12089475 TI - Molecular cloning of growth hormone encoding cDNA of Indian major carps by a modified rapid amplification of cDNA ends strategy. AB - A modified rapid amplification of cDNA ends (RACE) strategy has been developed for cloning highly conserved cDNA sequences. Using this modified method, the growth hormone (GH) encoding cDNA sequences of Labeo rohita, Cirrhina mrigala and Catla catla have been cloned, characterized and overexpressed in Escherichia coli. These sequences show 96-98% homology to each other and are about 85% homologous to that of common carp. Besides, an attempt has been made for the first time to describe a 3-D model of the fish GH protein. PMID- 12089479 TI - The gathering of evidence, evidence based medicine, and Joseph Heller's Catch 22. PMID- 12089480 TI - Symposium of breast imaging. PMID- 12089481 TI - Functional breast imaging using the single photon technique. AB - Early diagnosis of carcinoma of the breast is currently attempted by mammography and physical examination which together have a sensitivity of 85%. Mammography has a positive predictive value of only 15 to 40 percent. The usefulness of scintimammography for the detection of carcinoma of the breast has been studied which demonstrated improved specificity compared to conventional mammography. In future, with the development or dedicated breast imager and new targeted radiopharmaceuticals, we can achieve detection of smaller than 1 cm carcinoma. This type of imaging enables us to detect cancer at its 'pre-anatomic' stage. PMID- 12089482 TI - Positron emission tomography for the diagnosis of breast cancer. AB - This article reviews the literature on breast imaging with [18F]fluorodeoxyglucose positron emission tomography (FDG PET). In clinical applications, there is currently no defined role for detecting primary breast cancer. The limited sensitivity of FDG PET does not allow the exclusion of malignancy, in particular small breast carcinomas, micrometastases and small, tumour infiltrated lymph nodes. However, in advanced stages, PET accurately determines the extent of disease, including the loco-regional lymph node status. Furthermore, whole-body PET imaging promises a high diagnostic accuracy for detecting recurrent or metastatic breast carcinoma. Future clinical applications may include monitoring therapeutic effects. PMID- 12089483 TI - Monitoring therapy in breast cancer. AB - Breast cancer remains one of the most common cancers in the developed world. New treatments are proving effective against both limited disease and metastases. Nuclear medicine is in a unique position as it is one of the only methods used to image the breast which is linked to cell cycle changes, the receptors on the cell surface and the cells' response to chemotherapy. Nuclear medicine is unaffected by the anatomical changes seen post-chemotherapy and radiotherapy and is uniquely placed to become a major methodology in the continued assessment of the breast cancer patient. However, before this can happen the utility of nuclear medicine techniques must be proved in multi-centre trials. PMID- 12089484 TI - Innovation in early breast cancer surgery: radio-guided occult lesion localization and sentinel node biopsy. AB - The surgical management of non-palpable breast lesions remains controversial. At the European Institute of Oncology we have introduced a new technique, radio guided occult lesion localization (ROLL) to replace standard methods and overcome their disadvantages. Regarding axillary dissection, probe-guided biopsy of the sentinel node (SN) is easy to apply, and the whole procedure is associated to a low risk of false negatives. We suggest that the SN technique should be widely adopted to stage the axilla in patients with breast cancer with clinically negative lymph nodes. Large-scale implementation of the sentinel node technique will reduce the cost of treatment as a result of shorter hospitalization times. PMID- 12089485 TI - The value of 24-hour images after rest thallium injection. AB - Rest (201) Tl imaging has been used for detecting viability, but the ideal timing for imaging after injection to maximally estimate viability is not well established. Thirty patients with fixed or incompletely reversible defects on 4 h redistribution SPECT imaging after thallium rest injection underwent 24 h imaging. Global redistribution was subjectively rated none, minimal or meaningful by two experienced observers. Fourteen patients had no meaningful redistribution at either 4 h or 24 h. Ten patients had meaningful redistribution at 4 h only. Six patients had no meaningful redistribution at 4 h but did at 24 h. Defect size was quantified using a 70% threshold. For the total group, defect size was smaller at 4 h compared to immediate imaging (38+/-18% vs 41+/-19%, P=0.06) and smaller still at 24 h (36+/-16% vs 38+/-18%, P=0.02). Later (24 h) redistribution images detected additional redistribution in 30% of the patients who did not have meaningful redistribution on early (4 h) images, and in 8% of the segments which were abnormal at 4 h. It is concluded that, in patients who have incompletely reversible defects on early redistribution imaging at 4 h, late redistribution imaging after 24 h will demonstrate additional redistribution in 30% of the patients. PMID- 12089486 TI - Validation of left ventricular function from gated single photon computed emission tomography by using a scintillator-photodiode camera: a dynamic myocardial phantom study. AB - A scintillator-photodiode camera is able to acquire single photon emission computed tomography (SPECT) images by using a rotating chair system. We validated the left ventricular (LV) parameters of this camera system utilizing a dynamic myocardial phantom. Gated myocardial SPECT of a dynamic myocardial phantom (Hokkaido University type; end diastolic volume (EDV), 143 ml; end systolic volume (ESV), 107 ml; ejection fraction (EF), 25%) was performed with this scintillation camera. LV parameters were calculated using pre-installed software (Mirage Myocardial Perfusion SPECT) (study 1) and the other software (QGS; Cedars Sinai) (study 2). For comparison, SPECT from a traditional Anger camera were processed by the QGS software (study 3). The estimated volumes were similar among the three studies (EDV, 110+/-8 ml in study 1, 112+/-2 ml in study 2 and 111+/-1 ml in study 3; ESV, 86+/-8 ml in study 1, 93+/-4 ml in study 2 and 91+/-2 ml in study 3). The estimated EFs were 23+/-3%, 17+/-2%, and 18+/-1%, respectively. The calculated volume within each study was underestimated by approximately the same degree. However, each estimated EF value for each study was close to the actual value. The estimated LV function using the scintillator-photodiode camera system may be considered as a suitable alternative to the traditional Anger camera system. PMID- 12089487 TI - The use of [18 F]fluorodeoxyglucose positron emission tomography to differentiate between synovitis, loosening and infection of hip and knee prostheses. AB - Pain is a common unspecific symptom in orthopaedic prosthetics. The accurate differentiation between synovitis, loosening or infection is often difficult with conventional X-rays, arthrography or bone scintigraphy. Because of the high glucose uptake of inflammatory cells, [18F]fluorodeoxyglucose (18F-FDG) is an appropriate tracer for the evaluation of suspected inflammation or infection. In this preliminary study we describe 18F-FDG PET findings in patients referred for evaluation of painful hip or knee prostheses. We studied 23 patients with 28 prostheses, 14 hip and 14 knee prostheses, who had a complete operative or clinical follow-up. 18F-FDG PET scans were obtained with an ECAT EXACT HR+ PET scanner. High glucose uptake in the bone prostheses interface was considered as positive for infection, an intermediate uptake as suspect for loosening, and uptake only in the synovia was considered as synovitis. The imaging results were compared with operative findings or clinical outcome. PET correctly identified three hip and one knee prostheses as infected, two hip and two knee prostheses as loosening, four hip and nine knee prostheses as synovitis, and two hip and one knee prostheses as unsuspected for loosening or infection. In three patients covered with an expander after explantation of an infected prosthesis PET revealed no further evidence of infection in concordance with the clinical follow up. PET was false negative for loosening in one case. Our preliminary results suggest that FDG PET could be a useful tool for differentiating between infected and loose orthopaedic prostheses as well as for detecting only inflammatory tissue such as synovitis. PMID- 12089488 TI - Comparison of 99mTc ciprofloxacin, 99mTc white blood cell and three-phase bone imaging in the diagnosis of hip prosthesis infections: improved diagnostic accuracy with extended imaging time. AB - The purpose of this study was to compare the utility of 99mTc labelled ciprofloxacin (Infecton) imaging with the 99mTc white blood cell and three-phase bone imaging procedures for identifying hip prosthesis infection. We studied 30 symptomatic patients in whom infection was confirmed in eight and excluded in 22 cases based on clinical and microbiological findings. 99mTc ciprofloxacin images were obtained at 1, 4 and 24 h after the injection of the tracer, and the data were compared to those obtained from 99mTc leukocyte and three-phase bone imaging. The 99mTc ciprofloxacin imaging correctly identified all true infections. In 13 (59%) of the non-infected patients, non-specific uptake of 99mTc ciprofloxacin was found in the 1-h and 4-h images, which disappeared, however, in the 24-h images. When the early and late 99mTc ciprofloxacin images were compared, the specificity was found to improve from 41% to 95%, positive predictive value from 38% to 89%, and the diagnostic accuracy from 57% to 97%. The accuracy of the conventional 99mTc leukocyte imaging was 90%. Dynamic bone imaging also yielded abnormal findings in all the infected patients although also in 23% of the non-infected patients. Current data indicate that 99mTc ciprofloxacin is a useful method for confirming hip prosthesis infection. The diagnostic efficiency of this method is improved when the imaging time is extended to 24 h post-injection of the tracer. PMID- 12089489 TI - Utility of follow-up studies using meta-[123 I]iodobenzylguanidine scintigraphy for detecting recurrent neuroblastoma. AB - Neuroblastomas sometimes recur after the initial disappearance of the tumour. We evaluated the utility of meta-[123I]iodobenzylguanidine (123I-MIBG) scintigraphy for the detection of recurrent neuroblastomas by comparing with the measurement of biochemical markers and clinical findings. Forty patients who had received treatment for neuroblastomas were included in the study. After the disappearance of the initial tumours, periodic measurements of urinary vanillyl mandelic acid, homovanillic acid and serum neuron specific enolase values, and an 123I-MIBG scintigraphy were performed. Whenever an abnormal finding was observed, other appropriate examinations and/or follow-up examinations were performed to elucidate the true state of the patient. Eleven recurrent episodes in eight patients were observed. Most of them occurred in the bone marrow or bone. Corresponding symptoms were observed in only two episodes; the other episodes were asymptomatic, and discovered by the periodic examinations. 123I-MIBG scintigrams visualized the recurrent tumours in 10 (91%) episodes. Elevated tumour markers were observed in only three episodes. 123I-MIBG scintigrams visualized most of the recurrent tumours, unless they were accompanied by any symptoms or elevations in biochemical tumour markers. Periodic examinations with 123I-MIBG scintigraphy appears to be a useful technique for the detection of the recurrences. PMID- 12089490 TI - The role of IDA scintigraphy in the follow-up of liver disease in patients with cystic fibrosis. AB - The main aim of this study was to investigate the role of N-(2,4,6 trimethyl-3 bromophenylcarbamoylmethyl) iminodiacetic acid (IDA; Mebrofenin) scintigraphy in follow-up assessments of the biliary system in patients with cystic fibrosis associated liver disease. Fourteen patients from a study published in 1996 were re-examined after a mean interval of 4.7 years from their initial study, in which diisopropylphenyl carboxymethyl iminodiacetic acid (DISIDA) was used. The results of ultrasound, liver function tests and clinical examination were also compared. Twelve of the patients had been treated with ursodeoxycholic acid and taurine in the interim. Five subjects' IDA examinations showed a slight improvement on follow-up, six deteriorated, two were unchanged, whilst one demonstrated a 'mixed picture'. Overall, nine patients deteriorated in one or more of the tests. No patient showed a decline in all four investigations and only two in three. There was poor correlation between the various follow-up examinations, with different patients showing a decline in some tests but not others. This may be due to the mixture of functional and anatomical studies utilized, their differing sensitivies, and the fact that deterioration in one did not necessarily affect another. In conclusion, follow-up of hepatobiliary disease in patients with cystic fibrosis cannot be encompassed by one method alone. If early detection of disease progression would affect management, patients will continue to require a number of investigations rather than a single test. PMID- 12089491 TI - Clinical outcome of radiosynoviorthesis: a meta-analysis including 2190 treated joints. AB - A variety of indications for radiosynoviorthesis have been reported in literature, but the clinical outcome differs and depends on the primary disease and the pre-existing degenerative changes. This metaanalysis was carried out to establish groups for radiosynoviorthesis based on clinical outcome with respect to primary disease, clinical stage and the pre-existing degenerative changes. The literature search was carried out using the MEDLINE search term 'radionuclide synovectomy'. Based on reports in the literature we determined groups for radiosynoviorthesis for clinical use. Our literature list comprised 2190 joints that were treated with radiosynoviorthesis. The overall response rate for all treated joints was 72.5+/-17%. The mean improvement rate for the treated joints in rheumatoid arthritis was 66.7+/-15.4%. For osteoarthritis the success rate was 56+/-11%, with better results in case of minimal radiological changes. Radiosynoviorthesis in patients with changes according to Steinbrocker I and II was successful in 72.8+/-12.3%, and in 64+/-17.3%, respectively. Steinbrocker III and IV had a mean success rate of 52.4+/-23.6%. In the case of haemophilia and Willebrand's disease a reduction of joint bleedings and factor usages after radiosynoviorthesis was evident in 91+/-4.3%. In patients with pigmented villonodular synovitis radiosynoviorthesis was successful in 77.3+/-25.3%. It is concluded that radiosynoviorthesis provides better results in rheumatoid arthritis than in osteoarthritis. Minimal or moderate changes according to Steinbrocker stages I and II respond better to radionuclide therapy than do stages III and IV. Deformed or unstable joints might fail treatment and therefore surgical interventions should be considered. Close cooperation with orthopaedists and rheumatologists is necessary to consider radiosynoviorthesis in each patient to ensure optimal medical care. PMID- 12089492 TI - A class of asymmetrical nitrido 99mTc heterocomplexes as heart imaging agents with improved biological properties. AB - Asymmetrical heterocomplexes containing a terminal technetium-nitrogen multiple bond coordinated to one diphosphine ligand (PNP) and one dithiocarbamate ligand (DBODC), were obtained through a simple two-step procedure under controlled conditions. The resulting complexes [99mTc(N)(PNP)(DBODC)]+ are monocationic, and possess a distorted square-pyramidal geometry where the Tc triple bond N multiple bond occupies an apical position and the diphosphine and dithiocarbamate ligands span the residual four coordination positions on the basal plane through the two phosphorus atoms and the two sulfur atoms, respectively. Biodistribution data in rats demonstrated that these complexes were rapidly extracted by the myocardium, and retained in this region for a prolonged time. After a few minutes post injection, lung uptake became negligible, and liver washout was extremely rapid and quantitative. Analysis of heart/liver uptake ratios for these complexes revealed that their values increased exponentially in time, and after 60 min post injection liver activity was almost completely eliminated into the intestine. Comparison with heart/liver ratios determined for 99mTc sestamibi and 99mTc tetrfosmin showed that values for these latter compounds were approximately 10 times lower than those measured for [99mTc(N)(PNP)(DBODC)]+ complexes at 60 min post-injection. In conclusion, the monocationic tracers [99mTc(N)(PNP)(DBODC)]+ exhibit high myocardial uptake in rats and dramatically high heart/lung and heart/liver ratios, suggesting that this novel class of perfusion agents could be conveniently employed to obtain heart images with superior imaging quality. PMID- 12089493 TI - Uptake of cis-4-[18 F]fluoro-L-proline in scar formation: a marker of collagen synthesis? AB - Excessive scar formation is accompanied by abnormal collagen synthesis. The feasibility of monitoring collagen synthesis in vivo with no-carrier-added cis 4[18F]fluoro-L-proline (cis-FPro) was evaluated in an animal model with scar formation induced by implanted meshes. The abdominal wall of rats was replaced by alloplastic meshes. At days 3, 7, 14, 21 and 90 after implantation, the uptake of cis-FPro at 4 h post-injection was determined for resected samples of the mesh and normal tissues. The highest uptake was found in the kidneys (1.73+/ 0.47%ID/g) followed by the liver (0.59+/-0.19%ID/g). The meshes showed the maximum uptake at day 3 (0.20+/-0.07%ID/g) with a decrease to 0.10+/-0.03%ID/g at day 90 (P<0.001). After 3 days no connective tissue was shown by histopathological morphometric analysis. The maximum partial volume (PV%) of connective tissue was 43+/-14 PV% 90 days after implantation. The maximum levels of granulocytes and inflammatory infiltrate were found at day 3 with minimal levels at day 90, paralleling the course of cis-FPro uptake. In conclusion, the uptake of cis-FPro at 4 h post-injection is not related to the content of connective tissue. Cis-FPro radiolabelled with 18F appears not to be a suitable radiopharmaceutical for in vivo monitoring of collagen synthesis in scar formation. PMID- 12089494 TI - Orthopedic concerns in children with endocrine disorders. PMID- 12089495 TI - Sonographic investigation of anatomical specimens of infant hip joints. AB - The anatomical foundations of infant hip sonography techniques are ill-defined. We investigated anatomical specimens of infant hip joints in a water bath, with Graf's and Terjesen's methods. Acetabular position was varied in defined increments, with respect to the ultrasound beam. The alpha angles and the femoral head coverage were measured. Plastic acetabular casts were sawn along the sonographic section planes, and the cut sections compared with the sonographic sections. For images to be obtained, which were analysable by the two methods, the ultrasound beam had to intersect with the acetabular inlet plane at defined angles. The acetabular notch had to be anteriorly rotated from the ultrasound beam plane by at least 20 degrees. Beam entry within a 50 degrees sector posterior to the perpendicular on the inlet plane gave analysable images. The alpha angles and femoral head coverage were much affected by coronal-plane transducer tilt. Caudad tilts were associated with lesser values, a fact that should be borne in mind in clinical ultrasound investigations. PMID- 12089496 TI - Ultrasound screening of the infant's hip: introduction of a new combined angle. AB - This study evaluates the most widely used sonographic measurements for judging an infant's hip morphology. The alpha and beta angles of Graf, the bony coverage percentage of Morin and a new combined angle were analysed. This latter angle records the direction of the whole acetabulum (cartilagenous and bony) as one unit. One-hundred sonograms were examined independently by two observers to calculate the interobserver and intraobserver limits of agreement. The new combined angle proved to be the most reproducible measurement. Graf's alpha angle was the second most reliable measurement followed by the bone coverage percentage. The relationship between the combined angle and alpha angle was statistically significant with a correlation coefficient of 0.70 (P<0.001). PMID- 12089497 TI - Risk factors for developmental dysplasia of the hip: ultrasonographic findings in the neonatal period. AB - The relationship between ultrasonographic findings at birth and risk factors for developmental dysplasia of the hip have not been prospectively evaluated. Since implementing a routine screening programme for all new-born babies in 1989 we have collected 48 000 sets of data, including family history, birth presentation, mode of delivery and birth weight. Of the 92 babies (three per 1000 live births) with persistent ultrasonographic abnormality at 6 weeks only 20% displayed evidence of clinical instability at the original examination. Female babies without the additional risks of breech birth or positive family history were quantitatively the most significant group, accounting for 75% of cases treated. The majority of babies requiring intervention would not have been identified utilizing present criteria for selective ultrasound screening. PMID- 12089498 TI - Complications associated with surgically treated hip fractures in children. AB - Although hip fractures make up less than 1% of all children's fractures, they commonly produce late complications such as avascular necrosis, non-union, coxa vara, and premature physeal closure. A retrospective review of 17 patients with a mean age of 11 years (range, 7-14) and with a mean follow-up of 6 years (range, 2.5-12) who underwent open reduction and internal fixation for hip fractures was performed. There were 10 transcervical and five cervicotrochanteric fractures, one transepiphyseal fracture, and one intertrochanteric fracture. A total of 25 operations were performed. Overall, the complication rate was 59%, with some patients having more than one complication. Seven patients healed without any complications or sequelae. In the remaining 10 patients, the complications were avascular necrosis (nine patients), coxa vara (eight patients), premature physeal closure (seven patients), non-union (four patients), and chondrolysis (one patient). Avascular necrosis, non-union, and chondrolysis were associated with a poor outcome. At final follow-up, the results were assessed using Ratliff's criteria and included seven patients with good, six with fair, and four with poor outcome. PMID- 12089499 TI - A comparison of the biomechanical effects of opening and closing wedge varus osteotomies in Perthes' disease. AB - Varus femoral osteotomy is a commonly employed surgical method of effecting containment in the management of Perthes' disease. This can be achieved by performing either an opening lateral wedge or a closing medial wedge osteotomy. The effects of these two techniques on the biomechanical interrelationships about the involved hip were compared using a three-dimensional computer simulation. Both procedures lead to femoral shortening and reduce the moment capacity of the hip abductors. The shortening effect and consequent leg length discrepancy are greater for the closing wedge osteotomy than for the opening wedge procedure. Pelvic obliquity compensation, likely to be greater for a closing wedge osteotomy, provides a beneficial increase in femoral head coverage but at the cost of a further reduction in the hip abductor moment capacity. PMID- 12089500 TI - Ankle joint range of motion measurements in spastic cerebral palsy children: intraobserver and interobserver reliability and reproducibility of goniometry and visual estimation. AB - The aim of this study was to assess the intra- and interobserver reliability and reproducibility of goniometry and visual estimation of ankle joint range of motion measurements in children with spastic cerebral palsy. Forty-six ankles of 24 spastic cerebral palsy children were measured under a strict protocol. The global mean measurement error was 5 degrees (SD, 5 degrees) for intra- and interobserver measurements and 3 degrees (SD, 3 degrees) for goniometry versus visual estimation. Statistical analysis showed a high reliability for intra- and interobserver measurements (r>0.75), between visual estimation and goniometry (correlation coefficient, r>0.967; concordance coefficient, r>0.957). Both visual estimation and goniometry ankle range-of-motion measurements are reliable and reproducible in spastic cerebral palsy children if a strict but simple protocol is applied. PMID- 12089501 TI - Osteoid osteoma of the elbow in children: a report of three cases and a review of the literature. AB - Although osteoid osteoma is a relatively common lesion, it is rarely found at the elbow. We report three cases of osteoid osteoma of the olecranon fossa in patients under the age of 15 years. Diagnosis was delayed because of nonspecific clinical and radiological features. The three patients suffered from synovitis due to flexion contracture while at the same time prosupination remained normal. Only one patient complained of specific nocturnal pain. All cases had a latency between the onset of symptoms and the appearance of radiological signs. Open surgical excision of the nidus resulted in relief of pain in all cases and motion recovery in two cases. Diagnostic difficulties and treatment options are discussed. PMID- 12089502 TI - Rib fixation for humeral lengthening following interscapulothoracic resection for malignant tumor. AB - We report a case of humeral lengthening in an adolescent, 6.5 years after interscapulothoracic resection for an osteosarcoma of the scapula. The lengthening external fixator was augmented by ipsilateral rib fixation in order to prevent cephalad migration of the proximal fragment. At a 3-year follow-up, the patient was satisfied, with an improved cosmesis and function of his limb. PMID- 12089503 TI - Fractures of the olecranon during growth: a 15-25-year follow-up. AB - Ten girls and 13 boys with mean age 11 years when sustaining a fracture of the olecranon were examined at a mean of 19 years after the injury. Ten fractures were displaced less than 2 mm, three 2-3 mm, eight more than 3 mm and two were multifragmental. The treatment consisted of mobilization in three cases, plaster treatment in nine and open reduction and internal fixation in 11 cases. At follow up, 21 children had no subjective complaints while two had occasional mild pain. The upper arm circumference was thinner in the former injured extremities than in the uninjured (P<0.05). No other objective deficits were found. None had developed non-union or elbow osteoarthritis. Olecranon fractures during growth have an excellent long-term outcome. PMID- 12089504 TI - The treatment outcome of trigger thumb in children. AB - Our objectives were to determine the treatment outcome of trigger thumb and to identify its possible cause. One hundred and thirty-eight thumbs from 115 children were reviewed. There was an overall success rate of 66% following conservative treatment and the success rate appears to be higher in the younger age group and in those treated with splint therapy, which includes wearing a splint and regular therapy exercises. The outcome of the children who underwent surgery was good with a recurrence rate of 1.4% and a wound infection rate of 2.8%. Our results suggest that a more conservative approach to this problem should be adopted. Our data also suggest that this condition may be acquired rather than congenital. PMID- 12089505 TI - Genu recurvatum in an adolescent girl: hypothetical etiology and treatment considerations. A case report. AB - Genu recurvatum may be the result of various bony or soft tissue deformities around the knee. We present a case of genu recurvatum with radiological features similar to those seen in infantile fibrocartilagenous dysplasia. The options for surgical treatment are described and discussed. PMID- 12089506 TI - Malleolus externus plasty for joint reconstruction in fibular aplasia: preliminary report of a new technique. AB - The instability of the upper and the lower ankle joint represents a serious problem in patients with fibular aplasia. None of the previous techniques demonstrated at least sufficient results. In this report we present a new technique of malleolus externus plasty applied in a 5 1/2-year-old boy with fibular aplasia (type II according to Coventry and Johnson or type Ib according to Achtermann and Kalamchi) with a 6 cm length deficiency of the lower leg and 1 cm of the upper leg. Furthermore, he showed a tibial antecurvation deformity of 20 degrees, a fourth ray foot with adduction deformity of the hindfoot, cutane syndactyly D 2-4, and a hallux varus. This new technique works on the following principle. A triangular iliac crest transplant is implanted with an apophysis and the annexing fascia glutealis, including the osseous part of the transplant, in the lateral distal tibia in a way that the growing apophyseal part lays distally covering the lateral talus. Gluteal fascia annexed to the apophysis was used for the reconstruction of a lateral tendon. Using a ring fixator the transplant is fixed and coincidentally the lower leg lengthened and the axis corrected. The axis deformity and the leg-length deficiency of 7 cm were equalized. The fixator could be removed after 6 months. Radiologically, an entire integration of the iliac crest transplant was found. Magnetic resonance imaging showed it had sufficient circulation. The reexamination 2.5 years later demonstrated a simultaneous growth of the malleolus externus and the distal tibia and stable ankle joints with sufficient mobility and full weight-bearing capability. For support of the foot the boy uses ready-made shoes with curved arch supports. In conclusion, using this new technique an individually adapted lateral malleolus with growth-potential can be constructed that stabilizes the foot and the ankle joints. Thus, reluxations of the foot specifically in relation to lengthening of lower legs can be avoided with concurrent preservation of the range of movement of the ankle joint. PMID- 12089507 TI - Positive selection of MHC class Ib-restricted CD8(+) T cells on hematopoietic cells. AB - Unlike conventional CD8(+) T cells, major histocompatibility complex (MHC) class Ib-restricted CD8(+) T cells show an activated phenotype in uninfected mice and respond rapidly to foreign invaders. The underlying factors that contribute to these differences are not well understood. We show here that the activated phenotype of MHC class Ib-restricted CD8(+) T cells was partially acquired as a result of interactions in the thymus and reflected an increased capacity to be selected via interactions with MHC molecules on hematopoietic cells. Using bone marrow-chimeric mice, we have shown that MHC class Ib-restricted, but not MHC class Ia-restricted, CD8(+) T cells specific for Listeria monocytogenes were efficiently selected when MHC class I was expressed only on hematopoietic cells. Thus, the distinct functional properties of MHC class Ib-restricted versus MHC class Ia-restricted CD8(+) T cells may result, at least in part, from the different ways in which they are positively selected in the thymus. PMID- 12089508 TI - Disulfide exchange in domain 2 of CD4 is required for entry of HIV-1. AB - CD4, a member of the immunoglobulin superfamily of receptors that mediates cell cell interactions in the immune system, is the primary receptor for HIV-1. The extracellular portion of CD4 is a concatenation of four immunoglobulin-like domains, D1 to D4. The D1, D2 and D4 domains each contain a disulfide bond. We show here that the D2 disulfide bond is redox-active. The redox state of the thiols (disulfide versus dithiol) appeared to be regulated by thioredoxin, which is secreted by CD4(+) T cells. Locking the CD4 and the thioredoxin active-site dithiols in the reduced state with a hydrophilic trivalent arsenical blocked entry of HIV-1 into susceptible cells. These findings indicate that redox changes in CD4 D2 are important for HIV-1 entry and represent a new target for HIV-1 entry inhibitors. PMID- 12089509 TI - Origin of regulatory T cells with known specificity for antigen. AB - T cell receptor agonists can induce the differentiation of regulatory T (T(R)) cells. We report here that the immunoglobulin kappa-controlled expression of an agonist in different cell types correlated with the phenotype of the generated T(R) cells. We found that aberrant expression on thymic stroma yielded predominantly CD4(+)CD25(+) T(R) cells, which--under physiological conditions- may be induced by ectopically expressed organ-specific antigens and thus prevent organ-specific autoimmunity. Expression of the agonist antigen by nonactivated hematopoietic cells produced mostly CD4(+)CD25(-) T(R) cells. This subset can be derived from mature monospecific T cells without "tutoring" by other T cells and can be generated in the absence of a functioning thymus. Suppression of CD4(+) T cell proliferative responses by both CD25(+) and CD25(-) subsets was interleukin 10 (IL-10) independent and was overcome by IL-2. These data suggest that distinct pathways can be exploited to interfere with unwanted immune responses. PMID- 12089511 TI - HIV entry: are all receptors created equal? PMID- 12089510 TI - Fyn kinase initiates complementary signals required for IgE-dependent mast cell degranulation. AB - Fc epsilon RI activation of mast cells is thought to involve Lyn and Syk kinases proximal to the receptor and the signaling complex organized by the linker for activation of T cells (LAT). We report here that Fc epsilon RI also uses a Fyn kinase-dependent pathway that does not require Lyn kinase or the adapter LAT for its initiation, but is necessary for mast cell degranulation. Lyn-deficiency enhanced Fyn-dependent signals and degranulation, but inhibited the calcium response. Fyn-deficiency impaired degranulation, whereas Lyn-mediated signaling and calcium was normal. Thus, Fc epsilon RI-dependent mast cell degranulation involves cross-talk between Fyn and Lyn kinases. PMID- 12089512 TI - When public goes private. PMID- 12089513 TI - The ABCs of mitochondrial transcription. PMID- 12089515 TI - Reversing Babel with GO. PMID- 12089514 TI - Saving sulfur. PMID- 12089516 TI - Interfering with HIV. PMID- 12089518 TI - From QTL to gene: the harvest begins. PMID- 12089519 TI - The MHC and body odors: arbitrary effects caused by shifts of mean pleasantness. PMID- 12089522 TI - Large-scale prediction of Saccharomyces cerevisiae gene function using overlapping transcriptional clusters. AB - Genome sequencing has led to the discovery of tens of thousands of potential new genes. Six years after the sequencing of the well-studied yeast Saccharomyces cerevisiae and the discovery that its genome encodes approximately 6,000 predicted proteins, more than 2,000 have not yet been characterized experimentally, and determining their functions seems far from a trivial task. One crucial constraint is the generation of useful hypotheses about protein function. Using a new approach to interpret microarray data, we assign likely cellular functions with confidence values to these new yeast proteins. We perform extensive genome-wide validations of our predictions and offer visualization methods for exploration of the large numbers of functional predictions. We identify potential new members of many existing functional categories including 285 candidate proteins involved in transcription, processing and transport of non coding RNA molecules. We present experimental validation confirming the involvement of several of these proteins in ribosomal RNA processing. Our methodology can be applied to a variety of genomics data types and organisms. PMID- 12089521 TI - Genome-wide binding map of the histone deacetylase Rpd3 in yeast. AB - We describe the genome-wide distribution of the histone deacetylase and repressor Rpd3 and its associated proteins Ume1 and Ume6 in Saccharomyces cerevisiae. Using a new cross-linking protocol, we found that Rpd3 binds upstream of many individual genes and upstream of members of gene classes with similar functions in anabolic processes. In addition, Rpd3 is preferentially associated with promoters that direct high transcriptional activity. We also found that Rpd3 was absent from large sub-telomeric domains. We show by co-immunoprecipitation and by the high similarity of their binding maps that Ume1 interacts with Rpd3. In contrast, despite the known role of Ume6 in Rpd3 recruitment, only a limited number of the genes targeted by Rpd3 are also enriched for (or targeted by) Ume6. This suggests that Rpd3 is brought to many promoters by alternative recruiters, some of which may bind the putative cis-regulatory DNA elements that we have identified in sets of Rpd3 target genes. Finally, we show that comparing the genome-wide pattern of Rpd3 binding with gene expression and histone acetylation in the rpd3 Delta mutant strain reveals new sites of Rpd3 function. PMID- 12089523 TI - Reciprocal crossover asymmetry and meiotic drive in a human recombination hot spot. AB - Human DNA diversity arises ultimately from germline mutation that creates new haplotypes that can be reshuffled by meiotic recombination. Reciprocal crossover generates recombinant haplotypes but should not influence the frequencies of alleles in a population. We demonstrate crossover asymmetry at a recombination hot spot in the major histocompatibility complex, whereby reciprocal exchanges in sperm map to different locations in the hot spot. We identify a single-nucleotide polymorphism at the center of the hot spot and show that, when heterozygous, it seems sufficient to cause this asymmetry, apparently by influencing the efficiency of highly localized crossover initiation. As a consequence, crossovers in heterozygotes are accompanied by biased gene conversion, most likely occurring by gap repair, that can also affect nearby polymorphisms through repair of an extended gap. The result is substantial over-transmission of the recombination suppressing allele and neighboring markers to crossover products. Computer simulations show that this meiotic drive, although weak at the population level, is sufficient to favor eventual fixation of the recombination-suppressing variant. These findings provide an explanation for the relatively uniform widths of human crossover hot spots and suggest that hot spots may be generally prone to extinction by meiotic drive. PMID- 12089524 TI - Crossover clustering and rapid decay of linkage disequilibrium in the Xp/Yp pseudoautosomal gene SHOX. AB - Crossover between the human sex chromosomes during male meiosis is restricted to the terminal pseudoautosomal pairing regions. An obligatory exchange occurs in PAR1, an Xp/Yp pseudoautosomal region of 2.6 Mb, which creates a male-specific recombination 'hot domain' with a recombination rate that is about 20 times higher than the genome average. Low-resolution analysis of PAR1 suggests that crossovers are distributed fairly randomly. By contrast, linkage disequilibrium (LD) and sperm crossover analyses indicate that crossovers in autosomal regions tend to cluster into 'hot spots' of 1-2 kb that lie between islands of disequilibrium of tens to hundreds of kilobases. To determine whether at high resolution this autosomal pattern also applies to PAR1, we have examined linkage disequilibrium over an interval of 43 kb around the gene SHOX. Here we show that in northern European populations, disequilibrium decays rapidly with physical distance, which is consistent with this interval of PAR1 being recombinationally active in male meiosis. Analysis of a subregion of 9.9 kb in sperm shows, however, that crossovers are not distributed randomly, but instead cluster into an intense recombination hot spot that is very similar in morphology to autosomal hot spots. Thus, PAR1 crossover activity may be influenced by male-specific hot spots that are highly suitable for characterization by sperm DNA analysis. PMID- 12089525 TI - Mutant DNA-binding domain of HSF4 is associated with autosomal dominant lamellar and Marner cataract. AB - Congenital cataracts cause 10-30% of all blindness in children, with one-third of cases estimated to have a genetic cause. Lamellar cataract is the most common type of infantile cataract. We carried out whole-genome linkage analysis of Chinese individuals with lamellar cataract, and found that the disorder is associated with inheritance of a 5.11-cM locus on chromosome 16. This locus coincides with one previously described for Marner cataract. We screened individuals of three Chinese families for mutations in HSF4 (a gene at this locus that encodes heat-shock transcription factor 4) and discovered that in each family, a distinct missense mutation, predicted to affect the DNA-binding domain of the protein, segregates with the disorder. We also discovered an association between a missense mutation and Marner cataract in an extensive Danish family. We suggest that HSF4 is critical to lens development. PMID- 12089526 TI - Deletion in Catna2, encoding alpha N-catenin, causes cerebellar and hippocampal lamination defects and impaired startle modulation. AB - Mice homozygous for the cerebellar deficient folia (cdf) mutation are ataxic and have cerebellar hypoplasia and abnormal lobulation of the cerebellum. In the cerebella of cdf/cdf homozygous mice, approximately 40% of Purkinje cells are located ectopically in the white matter and inner granule-cell layer. Many hippocampal pyramidal cells are scattered in the plexiform layers, and those that are correctly positioned are less densely packed than are cells in wild-type mice. We show that fear conditioning and prepulse inhibition of the startle response are also disrupted in cdf/cdf mice. We identify a deletion on chromosome 6 that removes approximately 150 kb in the cdf critical region. The deletion includes part of Catna2, encoding alpha N-catenin, a protein that links the classical cadherins to the neuronal cytoskeleton. Expression of a Catna2 transgene in cdf/cdf mice restored normal cerebellar and hippocampal morphology, prepulse inhibition and fear conditioning. The findings suggest that catenin cadherin cell-adhesion complexes are important in cerebellar and hippocampal lamination and in the control of startle modulation. PMID- 12089528 TI - Self-inhibition of olfactory bulb neurons. AB - The GABA (gamma-aminobutyric-acid)-containing periglomerular (PG) cells provide the first level of inhibition to mitral and tufted (M/T) cells, the output neurons of the olfactory bulb. We find that stimulation of PG cells of the rat olfactory bulb results in self-inhibition: release of GABA from an individual PG cell activates GABA(A) receptors on the same neuron. PG cells normally contain high concentrations of intracellular chloride and consequently are depolarized by GABA. Despite this, GABA inhibits PG cell firing by shunting excitatory signals. Finally, GABA released during self-inhibition may spill over to neighboring PG cells, resulting in a lateral spread of inhibition. Given the gatekeeping role of PG cells in the olfactory network, GABA-mediated self-inhibition will favor M/T cell excitation during intense sensory stimulation. PMID- 12089527 TI - Ptprj is a candidate for the mouse colon-cancer susceptibility locus Scc1 and is frequently deleted in human cancers. AB - Only a small proportion of cancers result from familial cancer syndromes with Mendelian inheritance. Nonfamilial, 'sporadic' cancers, which represent most cancer cases, also have a significant hereditary component, but the genes involved have low penetrance and are extremely difficult to detect. Therefore, mapping and cloning of quantitative trait loci (QTLs) for cancer susceptibility in animals could help identify homologous genes in humans. Several cancer susceptibility QTLs have been mapped in mice and rats, but none have been cloned so far. Here we report the positional cloning of the mouse gene Scc1 (Susceptibility to colon cancer 1) and the identification of Ptprj, encoding a receptor-type protein tyrosine phosphatase, as the underlying gene. In human colon, lung and breast cancers, we show frequent deletion of PTPRJ, allelic imbalance in loss of heterozygosity (LOH) and missense mutations. Our data suggest that PTPRJ is relevant to the development of several different human cancers. PMID- 12089529 TI - Rapid recruitment of NMDA receptor transport packets to nascent synapses. AB - Although many of the molecules involved in synaptogenesis have been identified, the sequence and kinetics of synapse assembly in the central nervous system (CNS) remain largely unknown. We used simultaneous time-lapse imaging of fluorescent glutamate receptor subunits and presynaptic proteins in rat cortical neurons in vitro to determine the dynamics and time course of N-methyl-D-aspartate receptor (NMDAR) recruitment to nascent synapses. We found that both NMDA and alpha-amino 3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) subunits are present in mobile transport packets in neurons before and during synaptogenesis. NMDAR transport packets are more mobile than AMPAR subunits, moving along microtubules at about 4 microm/min, and are recruited to sites of axodendritic contact within minutes. Whereas NMDAR recruitment to new synapses can be either concurrent with or independent of the protein PSD-95, AMPARs are recruited with a slower time course. Thus, glutamatergic synapses can form rapidly by the sequential delivery of modular transport packets containing glutamate receptors. PMID- 12089531 TI - Slicing soup. PMID- 12089530 TI - Early mitochondrial calcium defects in Huntington's disease are a direct effect of polyglutamines. AB - Huntington's disease (HD) is caused by an expansion of exonic CAG triplet repeats in the gene encoding huntingtin protein (Htt), but the mechanisms by which this mutant protein causes neurodegeneration remain unknown. Here we show that lymphoblast mitochondria from patients with HD have a lower membrane potential and depolarize at lower calcium loads than do mitochondria from controls. We found a similar defect in brain mitochondria from transgenic mice expressing full length mutant huntingtin, and this defect preceded the onset of pathological or behavioral abnormalities by months. By electron microscopy, we identified N terminal mutant huntingtin on neuronal mitochondrial membranes, and by incubating normal mitochondria with a fusion protein containing an abnormally long polyglutamine repeat, we reproduced the mitochondrial calcium defect seen in human patients and transgenic animals. Thus, mitochondrial calcium abnormalities occur early in HD pathogenesis and may be a direct effect of mutant huntingtin on the organelle. PMID- 12089532 TI - US Supreme Court applies strict limits to patents. PMID- 12089533 TI - New SEC regulations could hinder biotech investment. US Securities and Exchange Commission. PMID- 12089534 TI - First biotech drug to treat psoriasis. PMID- 12089535 TI - India promotes GMOs in Asia. PMID- 12089536 TI - Bush domestic security proposal affects range of biotech programs. PMID- 12089538 TI - In silico biology through "omics". PMID- 12089539 TI - The need for national centers for proteomics. PMID- 12089540 TI - A summer financial makeover. PMID- 12089543 TI - Industry opposes genomic legislation. PMID- 12089544 TI - Validation of AAV-mediated gene targeting. PMID- 12089545 TI - Therapeutic cancer vaccines on trial. PMID- 12089546 TI - Transplanting cloned cells into therapeutic promise. PMID- 12089547 TI - Metabolic engineering by genome shuffling. PMID- 12089548 TI - Ribozyme programming extends the protein code. PMID- 12089549 TI - RNA as the catalyst for drug screening. PMID- 12089551 TI - The disparity between human cell senescence in vitro and lifelong replication in vivo. AB - Cultured human fibroblasts undergo senescence (a loss of replicative capacity) after a uniform, fixed number of approximately 50 population doublings, commonly termed the Hayflick limit. It has been long known from clonal and other quantitative studies, however, that cells decline in replicative capacity from the time of explantation and do so in a stochastic manner, with a half-life of only approximately 8 doublings. The apparent 50-cell doubling limit reflects the expansive propagation of the last surviving clone. The relevance of either figure to survival of cells in the body is questionable, given that stem cells in some renewing tissues undergo >1,000 divisions in a lifetime with no morphological sign of senescence. Oddly enough, these observations have had little if any effect on general acceptance of the Hayflick limit in its original form. The absence of telomerase in cultured human cells and the shortening of telomeres at each population doubling have suggested that telomere length acts as a mitotic clock that accounts for their limited lifespan. This concept assumed an iconic character with the report that ectopic expression of telomerase by a vector greatly extended the lifespan of human cells. That something similar might occur in vivo seemed consistent with initial reports that most human somatic tissues lack telomerase activity. More careful study, however, has revealed telomerase activity in stem cells and some dividing transit cells of many renewing tissues and even in dividing myocytes of repairing cardiac muscle. It now seems likely that telomerase is active in vivo where and when it is needed to maintain tissue integrity. Caution is recommended in applying telomerase inhibition to kill telomerase-expressing cancer cells, because it would probably damage stem cells in essential organs and even increase the likelihood of secondary cancers. The risk may be especially high in sun-exposed skin, where there are usually thousands of p53-mutant clones of keratinocytes predisposed to cancer. PMID- 12089552 TI - Historical claims and current interpretations of replicative aging. AB - Replicative aging is the process by which most normal human cells "count" the number of times they have divided, eventually undergoing a growth arrest termed cellular senescence. This process is dependent on the shortening of telomeres, repeated sequences at the ends of the chromosomes. The loss of telomeric sequences with each cell division eventually induces a growth arrest that has a similar phenotype to that of cells stressed by inadequate culture or other conditions. Experiments over the past several years have identified species in which replicative aging does not occur and many examples in which a failure to proliferate has been misinterpreted as replicative senescence. Insights from these studies now permit a reevaluation of much of the seemingly contradictory data concerning replicative aging. There are good theoretical reasons for believing a limited proliferative capacity contributes to declining tissue homeostasis with increasing age. Although the presence of telomere shortening provides strong circumstantial evidence that replicative aging is occurring in vivo, thus far there is only very limited direct evidence for actual physiological effects of replicative aging. PMID- 12089553 TI - Generation of histocompatible tissues using nuclear transplantation. AB - Nuclear transplantation (therapeutic cloning) could theoretically provide a limitless source of cells for regenerative therapy. Although the cloned cells would carry the nuclear genome of the patient, the presence of mitochondria inherited from the recipient oocyte raises questions about the histocompatibility of the resulting cells. In this study, we created bioengineered tissues from cardiac, skeletal muscle, and renal cells cloned from adult bovine fibroblasts. Long-term viability was demonstrated after transplantation of the grafts into the nuclear donor animals. Reverse transcription-PCR (RT-PCR) and western blot analysis confirmed that the cloned tissues expressed tissue-specific mRNA and proteins while expressing a different mitochondrial DNA (mtDNA) haplotype. In addition to creating skeletal muscle and cardiac "patches", nuclear transplantation was used to generate functioning renal units that produced urinelike fluid and demonstrated unidirectional secretion and concentration of urea nitrogen and creatinine. Examination of the explanted renal devices revealed formation of organized glomeruli- and tubule-like structures. Delayed-type hypersensitivity (DTH) testing in vivo and Elispot analysis in vitro suggested that there was no rejection response to the cloned renal cells. The ability to generate histocompatible cells using cloning techniques addresses one of the major challenges in transplantation medicine. PMID- 12089554 TI - Efficient mouse airway transduction following recombination between AAV vectors carrying parts of a larger gene. AB - The small packaging capacity of adeno-associated virus (AAV) vectors limits the utility of this promising vector system for transfer of large genes. We explored the possibility that larger genes could be reconstituted following homologous recombination between AAV vectors carrying overlapping gene fragments. An alkaline phosphatase (AP) gene was split between two such AAV vectors (rec vectors) and packaged using AAV2 or AAV6 capsid proteins. Rec vectors having either capsid protein recombined to express AP in cultured cells at about 1-2% of the rate observed for an intact vector. Surprisingly, the AAV6 rec vectors transduced lung cells in mice almost as efficiently as did an intact vector, with 10% of airway epithelial cells, the target for treatment of cystic fibrosis (CF), being positive. Thus AAV rec vectors may be useful for diseases such as CF that require transfer of large genes. PMID- 12089555 TI - In situ delivery of passive immunity by lactobacilli producing single-chain antibodies. AB - Lactobacilli have previously been used to deliver vaccine components for active immunization in vivo. Vectors encoding a single-chain Fv (scFv) antibody fragment, which recognizes the streptococcal antigen I/II (SAI/II) adhesion molecule of Streptococcus mutans, were constructed and expressed in Lactobacillus zeae (American Type Culture Collection (ATCC) 393). The scFv antibody fragments secreted into the supernatant or expressed on the surface of the bacteria showed binding activity against SAI/II in enzyme-linked immunosorbent assay (ELISA), and surface scFv-expressing lactobacilli agglutinated SAI/II-expressing S. mutans in vitro without affecting the corresponding SAI/II knockout strain. Lactobacilli expressing the scFv fragment fused to an E-tag were visualized by scanning electron microscopy (SEM) using beads coated with a monoclonal anti-E-tag antibody, and they bound directly to beads coated with SAI/II. After administration of scFv-expressing bacteria to a rat model of dental caries development, S. mutans bacteria counts and caries scores were markedly reduced. As lactobacilli are generally regarded as safe (GRAS) microorganisms, this approach may be of considerable commercial interest for in vivo immunotherapy. PMID- 12089556 TI - Genome shuffling of Lactobacillus for improved acid tolerance. AB - Fermentation-based bioprocesses rely extensively on strain improvement for commercialization. Whole-cell biocatalysts are commonly limited by low tolerance of extreme process conditions such as temperature, pH, and solute concentration. Rational approaches to improving such complex phenotypes lack good models and are especially difficult to implement without genetic tools. Here we describe the use of genome shuffling to improve the acid tolerance of a poorly characterized industrial strain of Lactobacillus. We used classical strain-improvement methods to generate populations with subtle improvements in pH tolerance, and then shuffled these populations by recursive pool-wise protoplast fusion. We identified new shuffled lactobacilli that grow at substantially lower pH than does the wild-type strain on both liquid and solid media. In addition, we identified shuffled strains that produced threefold more lactic acid than the wild type at pH 4.0. Genome shuffling seems broadly useful for the rapid evolution of tolerance and other complex phenotypes in industrial microorganisms. PMID- 12089557 TI - Genetic engineering of aminodeoxyhexose biosynthesis in Streptomyces fradiae. AB - The antibacterial properties of macrolide antibiotics (such as erythromycin, tylosin, and narbomycin) depend ultimately on the glycosylation of otherwise inactive polyketide lactones. Among the sugars commonly found in such macrolides are various 6-deoxyhexoses including the 3-dimethylamino sugars mycaminose and desosamine (4-deoxymycaminose). Some macrolides (such as tylosin) possess multiple sugar moieties, whereas others (such as narbomycin) have only single sugar substituents. As patterns of glycosylation markedly influence a macrolide's drug activity, there is considerable interest in the possibility of using combinatorial biosynthesis to generate new pairings of polyketide lactones with sugars, especially 6-deoxyhexoses. Here, we report a successful attempt to alter the aminodeoxyhexose-biosynthetic capacity of Streptomyces fradiae (a producer of tylosin) by importing genes from the narbomycin producer Streptomyces narbonensis. This engineered S. fradiae produced substantial amounts of two potentially useful macrolides that had not previously been obtained by fermentation. PMID- 12089558 TI - Protein-dependent ribozymes report molecular interactions in real time. AB - Most approaches to monitoring interactions between biological macromolecules require large amounts of material, rely upon the covalent modification of an interaction partner, or are not amenable to real-time detection. We have developed a generalizable assay system based on interactions between proteins and reporter ribozymes. The assay can be configured in a modular fashion to monitor the presence and concentration of a protein or of molecules that modulate protein function. We report two applications of the assay: screening for a small molecule that disrupts protein binding to its nucleic acid target and screening for protein protein interactions. We screened a structurally diverse library of antibiotics for small molecules that modulate the activity of HIV-1 Rev responsive ribozymes by binding to Rev. We identified an inhibitor that subsequently inhibited HIV-1 replication in cells. A simple format switch allowed reliable monitoring of domain-specific interactions between the blood-clotting factor thrombin and its protein partners. The rapid identification of interactions between proteins or of compounds that disrupt such interactions should have substantial utility for the drug-discovery process. PMID- 12089559 TI - A tRNA aminoacylation system for non-natural amino acids based on a programmable ribozyme. AB - The ability to recognize tRNA identities is essential to the function of the genetic coding system. In translation aminoacyl-tRNA synthetases (ARSs) recognize the identities of tRNAs and charge them with their cognate amino acids. We show that an in vitro evolved ribozyme can also discriminate between specific tRNAs, and can transfer amino acids to the 3' ends of cognate tRNAs. The ribozyme interacts with both the CCA-3' terminus and the anticodon loop of tRNA(fMet), and its tRNA specificity is controlled by these interactions. This feature allows us to program the selectivity of the ribozyme toward specific tRNAs, and therefore to tailor effective aminoacyl-transfer catalysts. This method potentially provides a means of generating aminoacyl tRNAs that are charged with non-natural amino acids, which could be incorporated into proteins through cell-free translation. PMID- 12089561 TI - Targeted transgene insertion into human chromosomes by adeno-associated virus vectors. AB - Efficient methods are needed for the precise genetic manipulation of diploid human cells, in which cellular senescence and low conventional gene targeting rates limit experimental and therapeutic options. We have shown previously that linear, single-stranded DNA vectors based on adeno-associated virus (AAV) could accurately introduce small (<20 bp) genetic modifications into homologous human chromosomal sequences. Here we have used AAV vectors to introduce large (>1 kb) functional transgene cassettes into the hypoxanthine phosphoribosyl transferase (HPRT) and Type I collagen (COL1A1) loci in normal human fibroblasts. The transgene cassettes are inserted at high frequencies (1% of the total cell population under optimal conditions) and without secondary mutations. Selection for the inserted transgene cassette can be used to enrich for targeting events, such that >70% of surviving cells have undergone gene targeting with an appropriately designed vector. This approach should prove useful both for functional genomic analysis in diploid human cells and for therapeutic gene targeting. PMID- 12089562 TI - Amine-modified random primers to label probes for DNA microarrays. AB - DNA microarrays have been used to study the expression of thousands of genes at the same time in a variety of cells and tissues. The methods most commonly used to label probes for microarray studies require a minimum of 20 microg of total RNA or 2 microg of poly(A) RNA. This has made it difficult to study small and rare tissue samples. RNA amplification techniques and improved labeling methods have recently been described. These new procedures and reagents allow the use of less input RNA, but they are relatively time-consuming and expensive. Here we introduce a technique for preparing fluorescent probes that can be used to label as little as 1 microg of total RNA. The method is based on priming cDNA synthesis with random hexamer oligonucleotides, on the 5' ends of which are bases with free amino groups. These amine-modified primers are incorporated into the cDNA along with aminoallyl nucleotides, and fluorescent dyes are then chemically added to the free amines. The method is simple to execute, and amine-reactive dyes are considerably less expensive than dye-labeled bases or dendrimers. PMID- 12089560 TI - Regulation of endogenous gene expression with a small-molecule dimerizer. AB - Artificial transcription factors containing designer zinc-finger DNA-binding domains (DBDs) have been used to activate or repress expression of a growing number of endogenous genes. We have combined targeted zinc-finger DBD technology with a dimerizer-regulated gene expression system to permit the small-molecule control of endogenous gene transcription. We constructed a dimerizer-responsive transcription factor that incorporates an artificial zinc-finger DBD targeted to the promoter of the human VEGF gene. Introduction of this activator into human cells allowed expression of the chromosomal VEGF gene to be induced by a small molecule dimerizer compound consisting of a nonimmunosuppressive rapamycin analog. We found that by directly regulating zinc-finger protein (ZFP) activity, we could circumvent difficulties encountered in the generation of cell lines stably expressing conventional unregulated activators. Dimerizer-dependent VEGF induction was rapid, tight, and dose dependent, and resulted in VEGF protein expression levels several-fold greater than those produced by the natural hypoxic response. PMID- 12089563 TI - Patenting bioinformatic inventions: emerging trends in the United States. PMID- 12089567 TI - International postdocs. PMID- 12089568 TI - Nurturing the business of biotechnology. PMID- 12089569 TI - The rise of the European bioentrepreneur. PMID- 12089570 TI - The engineers of change. PMID- 12089571 TI - Merlin's maestro. PMID- 12089572 TI - Private company profiles. PMID- 12089580 TI - Understanding business incubation. PMID- 12089581 TI - Innovation in Manchester. PMID- 12089582 TI - Incubation without walls. PMID- 12089583 TI - The nuts and bolts of startups. PMID- 12089584 TI - Writing your business plan. PMID- 12089585 TI - In-licensing as a business model. PMID- 12089586 TI - Partnering challenges for startups. PMID- 12089587 TI - Integrating the intellectual property value chain. PMID- 12089588 TI - Intellectual property as a foundation for funding. PMID- 12089589 TI - A counselor's dose of preventive medicine. PMID- 12089590 TI - Syndication--value in numbers. PMID- 12089591 TI - Strategies for managing risk in equity. PMID- 12089592 TI - Reinventing the biotech manager. PMID- 12089593 TI - Leadership management needs in evolving biotech companies. PMID- 12089594 TI - Dealing with the media. PMID- 12089595 TI - Chronic hypoxia induces prolonged angiogenesis in skeletal muscles of rat. AB - Skeletal muscle capillarity and fibre cross-sectional area were investigated within and between diaphragm (Diaph), extensor digitorum longus (EDL), soleus (SOL) and tibialis anterior (TA) muscles of control and chronic hypoxic (12 % O(2) for 6 weeks) adult male Wistar rats (final body mass approximately 355 g). Cryostat sections were stained for alkaline phosphatase activity to depict all capillaries, and for succinic dehydrogenase to demonstrate regional differences in oxidative capacity within the muscles. Hypoxia-induced angiogenesis occurred in all muscles (P < 0.01), with capillary-to-fibre ratio (C:F) being higher in the more active and oxidative muscles, Diaph (27 %) and SOL (26 %), than phasically active and glycolytic muscles, TA (21 %) and EDL (15 %). Diaph, SOL and EDL maintained fibre size, and hence showed an increased capillary density (CD) and reduced intramuscular diffusion distance (DD), whereas TA showed fibre hypertrophy and maintained CD and DD compared to control muscles. The extent of angiogenesis among different regions of muscle varied so as to suggest that muscle fibre size has an additional influence on capillary growth during chronic systemic hypoxia, which is progressive over an extended period of systemic hypoxia. PMID- 12089596 TI - Influence of genetically predisposed diabetes on ethanol-induced depression of cardiac contraction in adult rat ventricular myocytes. AB - Diabetes mellitus and alcohol (ethanol) intake are two positively correlated major risk factors for cardiovascular abnormalities. However, the interaction of the two on cardiac function is largely unknown. The purpose of the present study was to examine the impact of genetically predisposed diabetes on acute ethanol exposure-induced cardiac contractile depression at the myocyte level. Ventricular myocytes from spontaneously biobreeding diabetes-prone (BBDP) rats and their diabetes-resistant littermates (BBDR) were stimulated to contract at 0.5 Hz. Contractile properties analysed include: peak shortening amplitude (PS), time-to PS (TPS), time-to-90 % relengthening (TR(90)) and maximal velocities of shortening/relengthening (+/- dL/dt). BBDP rats displayed hyperglycaemia, reduced body weight gain and increased cardiac, hepatic and renal size. Myocytes isolated from BBDP rat hearts exhibited prolonged TPS and TR(90) associated with normal PS and +/- dL/dt, compared with myocytes from the BBDR group. Acute ethanol exposure (80-640 mg dl(-1)) caused a concentration-dependent inhibition of PS in both BBDR and BBDP myocytes. However, the degree of inhibition of PS was significantly reduced in BBDP myocytes compared to that of BBDR myocytes. The maximal inhibition was 52.9 % and 28.4 % in BBDR and BBDP groups, respectively. Ethanol significantly depressed +/- dL/dt in both BBDR and BBDP myocytes. In addition, ethanol did not affect TPS or TR(90) in either the BBDR or BBDP group. Collectively, these results suggest that the ethanol-induced depression in cardiac myocyte contraction may be 'shadowed' by genetically predisposed diabetes. PMID- 12089597 TI - Protein kinase C regulation of rat jejunal transport systems: mechanisms involved in bicarbonate absorption. AB - We examined whether protein kinase C (PKC) modulates the transport systems involved in bicarbonate movements across the plasma membranes of rat jejunum. Results of enzymatic assays provide evidence that under basal conditions conventional PKC (cPKC) is present in both basolateral membranes (BLMs) and apical (brush border) membranes (BBMs) of the enterocyte. In BLMs the basal expression of the kinase is low compared to expression in BBMs; however, treatment with Ca(2+) and phorbol 12-myristate 13-acetate (PMA) causes a significant increase, thus suggesting an asymmetrical kinase translocation. To explore the effect of PKC activation on membrane-bound transport mechanisms, 'in vitro' phosphorylated membrane vesicles were used to perform uptake studies. Results suggest that PKC activation exerts an inhibitory effect on the basolateral Cl(-)-HCO(3)(-) antiporter, whereas the basolateral HCO(3)(-) conductive pathway seems to be stimulated and Cl(-) conductance unaffected. The apical, but not basolateral, Na(+)-H(+) exchanger is inhibited by PKC activation. The specificity of the response to PKC was confirmed by using the kinase inhibitor staurosporine or the inactive phorbol ester 4-alpha-PMA. The inhibition of both apical Na(+)-H(+) and basolateral Cl(-)-HCO(3)(-) exchange activities suggests that the overall action of PKC causes a reduction of transepithelial bicarbonate transport. PMID- 12089598 TI - Ammonia inhibits sodium and chloride absorption in rat distal colon. AB - It was recently demonstrated that ammonia inhibits sodium absorption in the proximal colon of rats. In order to investigate the effect of luminal ammonia in the distal colon, sodium and chloride transport were measured in Ussing chambers. Under short-circuit conditions, distal colon absorbed sodium and chloride. When luminal ammonia (30 mmol l(-1)) was present, sodium and chloride absorption was diminished. Inhibition of the two Na(+)-H(+) exchanger isoforms NHE2 and NHE3, which are known to be located in the apical membrane of the distal colon epithelium, failed to influence the effect of ammonia on transepithelial sodium and chloride fluxes. The inhibitory effect of ammonia was eliminated under the following conditions: after block of carbonic anhydrases with acetazolamide, in the presence of an unspecific blocker of Na(+)-H(+) exchangers, and under chloride-free conditions. Ammonia did not alter electrogenic sodium absorption. These results demonstrate that luminal ammonia inhibits sodium and chloride absorption in rat distal colon. We suggest that ammonia inhibits NaCl absorption by interfering with a Na(+)-H(+) exchanger that is not NHE2 or NHE3 PMID- 12089599 TI - Effect of extracellular magnesium on nerve-mediated and acetylcholine-evoked in vitro amylase release in rat parotid gland tissue. AB - In this study the effects of changes in extracellular magnesium ([Mg(2+)](o)) and calcium ([Ca(2+)](o)) concentrations on basal and on nerve-mediated and acetylcholine (ACh)-evoked in vitro amylase release and calcium mobilization were investigated in rat parotid gland tissue. In the presence of a normal (2.56 mM) [Ca(2+)](o), both zero (0 mM) and an elevated (10 mM) [Mg(2+)](o) significantly attenuated basal and ACh-evoked amylase release compared to the response obtained in normal (1.1 mM) [Mg(2+)](o). During electrical field stimulation (EFS) of parotid tissues, only elevated [Mg(2+)](o) reduced amylase release. In a Ca(2+) free medium, both basal and ACh-evoked amylase output were markedly reduced compared to the responses obtained under similar conditions in normal [Ca(2+)](o). Again, the ACh-induced amylase release in a Ca(2+)-free solution was larger in normal [Mg(2+)](o) than when the [Mg(2+)](o) was either zero or was elevated to 10 mM. Perturbation of [Mg(2+)](o) had no significant effect on basal intracellular free calcium concentration ([Ca(2+)](i)) in parotid acinar cells loaded with the fluorescent Ca(2+) indicator fura-2. Both zero Mg(2+) and an elevated [Mg(2+)](o) significantly reduced the ACh-induced rise in the peak and the plateau phase of the Ca(2+) transient that was seen in normal [Mg(2+)](o). In parotid acinar cells loaded with the fluorescent Mg(2+) indicator magfura-2, ACh elicited a gradual decrease in intracellular free Mg(2+) concentration ([Mg(2+)](i)) to below the basal level. The results indicate that both hypo- and hypermagnesaemia may reduce both basal and ACh-evoked amylase secretion from the salivary gland. As far as the ACh-evoked response is concerned, the effect may be exerted by a decrease in cellular Ca(2+) transport. PMID- 12089600 TI - Angiotensin II type 1 (AT(1)) receptor blockade enhances the L-NAME-induced vasoconstriction in rat submandibular gland. AB - The vasoregulatory role of nitric oxide (NO) and angiotensin II type 1 (AT(1)) receptors in the circulation of the submandibular gland (SMG) of rats was studied. The glandular blood flow was determined by means of laser Doppler flowmetry and rubidium isotope technique. The data obtained by these two methods correlated well (r = 0.77; P < 0.01). The AT(1) receptor antagonist candesartan (0.5 mg kg(-1), i.v.) reduced the vascular resistance in the SMG by 37 % (P < 0.05). By contrast, the NO synthase blocker L-NAME (15 mg kg(-1), i.v.) significantly increased vascular resistance in the SMG both in candesartan treated (P < 0.001) and non-treated (P < 0.001) animals. The increase in resistance was greater (P < 0.05) after previous blockade of AT(1) receptors. These findings suggest that the AT(1) receptors have an important role in the vasoregulation of the SMG in the rat. As a result of AT(1) blockade, NO-dependent tone of glandular vessels may be enhanced significantly. PMID- 12089601 TI - Myocardial stunning reduces the effects of nitric oxide on coronary capillary perfusion in the rabbit. AB - This investigation tested the hypothesis that the effects of nitric oxide synthase on myocardial capillary perfusion were reduced during myocardial stunning. Anaesthetized open-chest rabbits were assigned to either a control group or a group treated with a nitric oxide synthase inhibitor. To induce myocardial stunning, a coronary artery was occluded (for 15 min) and then reperfused (for 15 min) twice. During reperfusion, rabbits were given either saline or N(G)-nitro-L-arginine methyl ester (L-NAME, 30 mg kg(-1)) followed by i.v. injection of 150 mg kg(-1) fluorescein isothiocyanate (FITC)-labelled dextran (molecular weight, 150,000) for 14 s. Fluorescence microscopy was used to identify the perfused vessels and an alkaline phosphatase stain was used to locate the total microvasculature. The 'closest-individual' method was used to estimate the geometric distribution of capillaries. No significant differences were observed in the total volume fraction (mm(3) of capillaries per mm(3) of tissue) between the control and stunned regions in either the saline- (0.045 +/- 0.008 and 0.042 +/- 0.009, respectively) or the L-NAME-treated hearts (0.060 +/- 0.010 and 0.049 +/- 0.005, respectively). There were no significant differences in the percentage volume fraction of perfused capillaries between the control and the stunned regions (49 +/- 4 % and 54 +/- 4 %, respectively) in saline-treated hearts. In hearts treated with L-NAME, the percentage of perfused capillaries was significantly reduced. The reduction was significantly greater in the control region (approximately 27 %) than the stunned region (approximately 17 %). Closest individual analysis of the perfused capillary distribution in both groups demonstrated a similar unchanged distribution. Thus, nitric oxide synthase is an important regulator of basal coronary capillary perfusion, and its effects are significantly reduced by myocardial stunning. PMID- 12089602 TI - The cardiovascular and renal effects of acute and chronic inhibition of nitric oxide production in fetal sheep. AB - The acute and long-term effects of blockade of nitric oxide (NO) production were studied in six chronically catheterised fetal sheep aged from 116 and 118 days; six untreated fetal sheep received injections of saline. Injection of 10 mg (kg maternal body wt)(-1) of the nitric oxide synthase (NOS) inhibitor N(omega)-nitro L-arginine (NOLA) to the fetus, caused an immediate rise in fetal mean arterial pressure (MAP, P < 0.005) and a reflex fall in fetal heart rate (FHR, P < 0.001). Plasma renin concentration (PRC) fell from 8.4 +/- 3.3 to 1.5 +/- 0.3 ng ml(-1) h(-1) (P < 0.001) and was dependent on MAP (P = 0.001). Glomerular filtration rate (GFR) tended to increase, but renal blood flow (RBF) velocity decreased (P < 0.001). Thus filtration fraction (FF) increased (P < 0.025). Urine flow and sodium excretion increased (P < 0.001 for both). Fractional sodium reabsorption decreased (P < 0.05). In fetuses treated with NOLA, arterial pressure was found to affect glomerular haemodynamics and renal tubular handling of sodium. No such relationships were observed in untreated fetuses. The vascular responses to acetylcholine tended to be less (P = 0.07) and the responses to noradrenaline were enhanced in NOLA-treated fetuses. There were no changes in untreated fetuses. Fetuses were then injected twice daily with either 5 mg kg(-1) NOLA or saline for the next 2 days. On the 4th day, injection of 10 mg kg(-1) NOLA did not have any effects on MAP, FHR or renal function. However, the pressor responses to angiotensin II (Ang II) were enhanced (P < 0.005), as was the response to noradrenaline but to a lesser extent. It is concluded that endothelial production of NO maintains normal fetal blood pressure, renal vascular resistance and fetal renal function. When NO production was blocked by repeated injections of NOLA, other vasodilator pathways took over the maintenance of cardiovascular and renal vascular tone. However, alterations in both cardiovascular and renal function were still present. That is, there was increased pressor sensitivity to exogenous Ang II and unmasking of effects of arterial pressure on glomerular and tubular function. PMID- 12089603 TI - The effects of pregnancy and maternal nutrition on the maternal renin-angiotensin system in sheep. AB - Physiological changes occurring in the mother during pregnancy can determine the outcome of pregnancy in terms of birthweight and neonatal viability. Maternal adaptations include plasma volume expansion linked to enhanced activity of the renin-angiotensin system (RAS). The present study was designed to determine whether these changes occur very early in gestation, and the extent to which maternal nutrient restriction may compromise the maternal RAS. Using sheep, we have investigated the effects of pregnancy per se, maternal nutrient restriction and later restoration of maternal diet on maternal body weight, plasma volume and plasma renin concentration (PRC), and angiotensinogen (Aogen) and arginine vasopressin (AVP) concentration. During the period of placental growth (i.e. 28 80 days gestation) ewes were fed either a nutrient-restricted (NR) diet or were well fed (WF). NR ewes consumed between 3.2 and 3.8 MJ day(-1) of metabolisable energy (ME) which is close to 60 % of requirements taking into account the ME required for both ewe maintenance and growth of the conceptus in order to produce a 4.5 kg lamb at term. WF ewes consumed 150 % of ME requirements. Restoration of maternal diet between 80 and 140 days gestation (i.e. fed to satiety and consuming between 8 and 10.9 MJ day(-1), which is close to 150 % of ME requirements) followed previous nutrient restriction. Between pre-conception and 28 days gestation, plasma volume increased in conjunction with a decline in PRC and Aogen concentration. During the period of nutrient restriction ewe body weight did not increase and plasma volume was lower in NR than WF ewes. During this time there was no effect of maternal nutrition on PRC; however, Aogen concentration was lower in the NR group. From 80 days gestation following the rise in food intake for previously NR ewes, greater increases in ewe body weight, plasma volume and PRC occurred up to term compared with ewes that were well fed throughout gestation. Plasma AVP concentration was not significantly affected by either maternal nutrition or gestational age. In conclusion, the stimulus of moderately severe maternal nutrient restriction evoked smaller rises in maternal weight, plasma volume and Aogen concentration than occurred in ewes that were well fed throughout gestation. Following the restoration of maternal diet after 80 days gestation, PRC gradually rose to peak at term. These adaptations in the maternal RAS during the critical period of placental growth may have long-term effects on fetal development. PMID- 12089604 TI - Retrograde transfer of ovarian steroid hormones to the ovary in the porcine periovarian vascular complex. AB - The aim of the present study was to investigate the mechanism of the retrograde transfer of ovarian steroid hormones from the ovarian lymphatic and venous effluent to the arterial blood supplying the ovary. In the first experiment, reproductive organs were collected from gilts in the luteal (n = 10) and follicular (n = 10) phase of the oestrous cycle. The ovary with the mesovarium was isolated and perfused through the ovarian artery with warmed, oxygenated autologous blood. The concentrations of progesterone and oestradiol in ovarian arterial blood increased on passing through the ovarian artery to the ovary, in the luteal phase, from 20.3 +/- 2.1 to 31.4 +/- 3.9 ng ml(-1) (P < 0.001) and from 6.2 +/- 0.8 to 11.4 +/- 1.4 pg ml(-1) (P < 0.001), respectively, and in the follicular phase, from 1.2 +/- 0.2 to 2.2 +/- 0.4 ng ml(-1) (P < 0.001) and from 8.2 +/- 1.8 to 13.2 +/- 2.3 pg ml(-1) (P < 0.001), respectively. Approximately 17.5 +/- 3.9 % of the progesterone and 12.6 +/- 1.7 % of the oestradiol found in the ovarian venous effluent was retrogradely transferred from the ovarian venous blood to the ovary in the luteal phase. In the follicular phase, these values were 10.1 +/- 2.0 % and 8.6 +/- 1.4 %, respectively. The efficiency of retrograde transfer of oestradiol and the rate of retrograde transfer of progesterone differed between phases of the oestrous cycle (P < 0.05 and P < 0.0001, respectively). A direct relationship between the concentration of the steroids in the venous effluent and the efficiency and rate of the retrograde transfer to the ovary was not found. In the second experiment (luteal phase, n = 10; follicular phase, n = 5), the concentration of progesterone and oestradiol increased in both ovarian arterial blood (P < 0.0001) and in the venous effluent (P < 0.0001) after administration of the steroids into the lymphatic vessels of the isolated mesovarium with separated ovary. In the third experiment (follicular phase, n = 5), with the mesovarium isolated after the ovary was removed and ovarian venous blood flowing out under the force of gravity (without the blood pressure in the ovarian vein), it was demonstrated that the veno-venous network covering the branches of the ovarian artery was supplied with the blood flowing out from the mesovarian tissue and that the filling of the veno-venous network was dependent on the blood pressure in the ovarian artery. We conclude that the effective retrograde transfer of steroid hormones from ovarian venous and lymphatic effluent to the ovary is accomplished not only by the classical counter-current exchange mechanism, but also as a result of complex processes that may be dependent on a specific part of the circulation of the blood and lymph in the periovarian vascular complex of the mesovarium. PMID- 12089605 TI - Hypoxia and expression of VEGF-A protein in relation to capillary growth in electrically stimulated rat and rabbit skeletal muscles. AB - To investigate the role of hypoxia as a stimulus to the early upregulation of vascular endothelial growth factor (VEGF) in fast skeletal muscles during chronic low frequency stimulation, blood flow, oxygen consumption, VEGF expression and capillary : fibre ratio were measured in chronically stimulated tibialis anterior and extensor digitorum longus (EDL) muscles in rabbits and rats. No differences were found in blood flow, oxygen consumption and extraction between rabbit muscles stimulated for 2 or 4 days (8 h on-16 h off) and controls. Muscle P(O(2)) polarographically measured immediately at the end of stimulation on day 2 was also no different from control under resting conditions (10.7 +/- 1.6 vs. 9.5 +/- 1.2 Torr, n.s.). Unlike control muscles, however, P(O(2)) in 2 day stimulated muscles did not increase immediately after a further acute bout of contractions. This difference was not apparent after similar acute contractions in 4 day stimulated muscles. The involvement of VEGF in early angiogenesis in stimulated muscles was studied in serial cryosections of rat EDL. The proportion of capillaries positively immunostained for VEGF increased from 25 +/- 1 % to 40 +/- 1 % (P < 0.05) in muscles removed on day 2 immediately at the end of chronic stimulation; it decreased slightly after 16 h rest, and increased again after 4 days of stimulation. Capillary : fibre ratio was unchanged throughout the experiment. Capillary cell proliferation increased only after the rest period on day 2 (20-fold increase) and day 4 (12-fold increase), indicating angiogenesis in progress. Thus the timing of transient hypoxia and increase in capillary-linked VEGF in stimulated muscles, albeit in different species, was similar, and increased VEGF staining and capillary cell proliferation occurred even after the hypoxia had resolved. This suggests (1) a connection between hypoxia and VEGF during the early stages of stimulation, although ensuing capillary proliferation may thereafter rapidly correct for local hypoxia, and (2) that the subsequent angiogenesis and VEGF expression are dependent on factors other than hypoxia. PMID- 12089606 TI - Changes in human muscle transverse relaxation following short-term creatine supplementation. AB - The rapid increase in body mass that often occurs following creatine (Cr) supplementation is believed to be due to intracellular water retention. The purpose of this study was to determine whether Cr consumption alters the magnetic resonance (MR) transverse relaxation (T(2)) distribution of skeletal muscle. Transverse relaxation can be used to model water compartments within a cell or tissue. In this double-blind study, subjects were asked to supplement their normal diet with creatine monohydrate (20 g day(-1) for 5 days) mixed with a grape drink (Creatine group, n = 7), or the grape drink alone (Placebo group, n = 8). Phosphorous MR spectroscopy was used to determine the effectiveness of the supplementation protocol. Subjects that responded to the Cr supplementation (i.e. showed a > 5 % increase in the ratio of the levels of phosphocreatine (PCr) and ATP) were placed in the Creatine group. Both proton MR imaging and spectroscopy were used to acquire T(2) data, at 1.89 T, from the flexor digitorum profundus muscle of each subject before and after supplementation. Following the supplementation period, the Creatine group showed a gain in body mass (1.2 +/- 0.8 kg, P < 0.05, mean +/- S.D.), and an increase in PCr/ATP ratio (23.8 +/- 16.4 %, P < 0.001). Neither group showed any changes in intracellular pH or T(2) calculated from MR images. However, the spectroscopy data revealed at least three components (> 5 ms) at approximately 20, 40 and 125 ms in both groups. Only in the Creatine group was there an increase in the apparent proton concentration of the two shorter components combined (+5.0 +/- 4.7 %, P < 0.05). According to the cellular water compartment model, the changes observed in the shorter T(2) components are consistent with an increase in intracellular water. PMID- 12089608 TI - [Early systemic sclerosis: diagnostic criteria and work-up]. PMID- 12089607 TI - Route of administration differentially affects fevers induced by Gram-negative and Gram-positive pyrogens in rabbits. AB - We have investigated the febrile responses of New Zealand White rabbits to a Gram negative pyrogen (bacterial lipopolysaccharide (LPS) from Salmonella typhosa), commonly associated with systemic infection, and a Gram-positive pyrogen (Staphylococcus aureus), more frequently associated with superficial soft tissue infection, each administered via one of four different routes (intravenous, intramuscular, subcutaneous or intraperitoneal) at each of three different doses (LPS: 0.1, 1 and 10 microg kg(-1); S. aureus: 1.5 x 10(7), 1.5 x 10(8) and 1.5 x 10(9) cell walls kg(-1)). Intravenous administration of LPS evoked rapid, dose dependent biphasic fever. Injection of LPS by the other routes also evoked dose dependent fever. However, these fevers were monophasic, had increased latency of onset, and were of lower amplitude. It is important to note that a dose of approximately 10 and 100 times that of the standard intravenous dose was required to produce a similar peak rise in temperature when administered subcutaneously and intraperitoneally, respectively. Intravenous injection of the highest dose of S. aureus evoked dose-dependent biphasic fever, with short latency of onset, which was very similar to that induced by intravenous LPS. At lower doses, intravenous S. aureus induced monophasic fever. No fever occurred when the same doses of S. aureus were administered by any other route. We conclude that any of the four routes may be used for the study of LPS-induced fever, provided that the doses are adjusted. However, studies of S. aureus-induced fever, and detection of contamination with either pyrogen, requires intravenous injection. PMID- 12089609 TI - [Role of chemokines in the pathogenesis of chronic synovitis during rheumatoid arthritis]. AB - Chemokines play a central role in the pathogenesis of rheumatoid arthritis (RA) synovitis which is characterised by new blood vessel formation, thickening of the lining layer and infiltration of immune cells. The inflammatory infiltrate is generated by a series of events which include the recruitment of leukocytes from the blood stream into the tissue, their local retention and activation to effector cells. All these processes are finely regulated by the interplay of different cell adhesion molecules (CAMs) and chemoattractant factors called chemokines (CK). CK are a superfamily of small proteins that play a crucial role in immune and inflammatory reactions. These chemoattractant cytokines share structural similarities including four conserved cysteine residues which form disulphide bonds in the tertiary structure of the proteins. CK mediate their effects by binding specific receptors (CK-R) characterised by a domain structure which spans the cell membrane seven times and signal through heterotrimeric GPT binding proteins. Activation of the CK network results in an amplification of the inflammatory cascade and consequently in the progressive destruction of RA joints. The recognition of the central role of CK in inflammation has paved the way to the development of new agents capable of interfering with CK and CK-R. This review will focus in particular on the role of CK in regulating leukocyte trafficking in RA joints. PMID- 12089610 TI - [TNF- alpha gene polymorphisms in rheumatoid arthritis patients treated with anti TNF-alpha agents: preliminary results]. AB - OBJECTIVES: To study -238 and +489 TNF-alpha polymorphisms in severe-unresponsive (more than 6 swollen joints and still active disease despite at least 6 months of DMARDs combination therapy) and mild-responsive (less than 3 swollen joints and good response to MTX or other conventional DMARDs) rheumatoid arthritis (RA). METHODS: We investigated 100 RA patients (56 with severe and 44 with mild disease activity) and 45 healthy blood donors (HBDs). Genotyping was performed by PCR restriction fragment length polymorphism procedure. Several clinical and serological parameters were also examined. RESULTS: Severe RA patients disclosed the -238 GG genotype in 100% of the cases versus 95.5% in the mild-responsive patients and 91.2% in the HBDs. The +489 GG genotype disclosed only a trend towards a prevalence in severe RA patients. However the +489 A allele seems to associates with early onset, longer disease duration and longer responsiveness to conventional therapy. CONCLUSION: The -238 AG genotype is absent in severe unresponsive RA, but present in mild-responsive RA subjects. Thus -238 GG homozygosity associates with severity and unresponsiveness. In contrast the +489 polymorphism does not segregate differently between responsive and unresponsive RA patients. PMID- 12089611 TI - [Seronegative spondyloarthropathies and allergic diseases in patients with ulcerative colitis]. PMID- 12089612 TI - [The high frequency of spontaneous micronuclei observed in lymphocytes of systemic sclerosis patients: preliminary results]. AB - OBJECTIVE: Aim of the study is to assess the presence of spontaneous chromosome damage in patients affected by limited (lSSc) or diffuse (dSSc) Systemic Slerosis, using the micronucleus (MN) assay. METHODS: We evaluated MN frequency in cultured peripheral lymphocytes of 18 SSc and in a group of 20 healthy controls. Patients were also classified as ACA+, Scl70+, FAN+ according to the presence of the specific anti-nuclear antibodies. We also explored the hypothesis that the extent of cytogenetic alteration might be related to the severity of the pathological condition and/or to the immunological profile. RESULTS: Compared to controls, the patient group as a whole showed significantly higher MN frequencies (10.8+/-4.5 vs. 27.8+/-13.7, p<0.001). No correlation was found between spontaneous chromosome damage and severity of the disease, being MN frequency 33.1+/-17.0 and 19.8+/-2.7 in lSSc and dSSc, respectively. Interestingly, ACA+ subjects displayed the highest MN frequency (36.9+/-15.0), as compared to patients with different antibody pattern (Scl70+, FAN+; 19.7+/-8.2). CONCLUSIONS: Our results confirm the presence of chromosomal damage in circulating lymphocytes of SSc patients and would suggest a key role of antibodies to the centromere in determining the observed cytogenetic anomalies. PMID- 12089613 TI - [Methodology of an epidemiologic prevalence study in rheumatology: the Chiavari study]. AB - OBJECTIVES: Goals of epidemiological studies are the description of the measures of frequency of diseases, the attempt to clarify possible etiopathogenic mechanisms, and the provision of data to support health policy decisions. To increase the familiarity of rheumatologists toward epidemiology, we describe the methodology used in a prevalence study of musculoskeletal complaints performed in Chiavari, Italy. METHODS: A questionnaire, originally developed by the Epidemiology Unit of the Arthitis Research Council in Manchester, UK, to investigate the prevalence of rheumatoid arthritis, was used after translation and validation. 4456 subjects aged 16 years or more listed in four general practices were invited to participate in the study and to fill the ARC questionnaire. The 3294 responders reported a) any past occurrence of joint swelling lasting more than four weeks and the distribution of the swollen joints on a mannequin; b) any joint pain lasting more than four weeks; c) current joint pain or swelling; d) morning stiffness; e) whether they had been previously told by a doctor they had arthritis. RESULTS: Four steps were necessary to obtain a 74% response, i.e. direct contact, two mailings and a phone interview. The performance of the different questions was good. The prevalence of the most common conditions among patients answering positively to the questions regarding morning stiffness and symmetrical swelling of joints was as follows: osteoarthritis 2.60%, fibromyalgia 1.30%, carpal tunnel syndrome 1.14%, rheumatoid arthritis 0.31%, and psoriatic arthritis 0.10%. CONCLUSIONS: Methodological issues regarding the selection of the population and sample to study, the development of a questionnaire, and the problems in obtaining valid informations are discussed. PMID- 12089614 TI - [Rhinopharyngeal carcinoma and dermatomyositis: description of a clinical case]. AB - Nasopharyngeal carcinoma has long been reported as the predominant type of cancer associated with dermatomyositis in many several Asian countries, including Hong Kong, Singapore, and Southern-Cina. Dermatomyositis is one of the idiopathic inflammatory myopathies showing characteristic cutaneous manifestations. Reviews from the western literature have demonstrated that certain cancers, such as ovarian and breast carcinoma in women and lung and prostate carcinoma in men, are highly associated with DM relative to the general population. We report the case of a Caucasian Italian patient with nasopharyngeal carcinoma and dermatomyositis. Considering the rarity of nasopharyngeal carcinoma among whites, both the detection and the report of each new case are noteworthy in defining the geographic and ethnic distribution of this tumor. PMID- 12089615 TI - [Anterior chest wall examination reviewed]. AB - Anterior chest wall involvement is not infrequently observed within inflammatory arthropaties, particularly if one considers seronegative spondyloarthritides and SAPHO syndrome. Physical examination is unreliable and conventional X-rays analysis is an unsatisfactory tool during diagnostic work-up of this region. Scintigraphic techniques yield informations both on the activity and on the anatomical extent of the disease while computerized tomography visualize the elementary lesions, such as erosions, which characterize the process. Moreover, when available, magnetic resonance imaging couple the ability to finely visualize such lesions with the possibility to show early alterations and to characterize the "activity" of the disease, presenting itself as a powerful tool both for diagnosis and follow-up. This review briefly shows the applications of imaging techniques for the evaluation of the anterior chest wall focusing on what has been done in the SAPHO syndrome which can be considered prototypical for this regional involvement since it is the osteo-articular target mainly affected by the disease. PMID- 12089616 TI - [Polyarticular heterotopic ossification complicating drug induced coma]. AB - The Authors describe a case of generalised heterotopic ossification (HO) complicating the course of a 31 years old male patient, with a history of Hodgkin's lymphoma and a of long staying in an intensive care unit, where he had been placed on artificial coma. The clinical features, as well as radiological findings and therapy, are discussed. PMID- 12089617 TI - [The "Hospital Real de San Josef de los Naturales" and the rheumatic conditions found in New Spain during the 16th century]. AB - The rheumatic conditions found in New Spain during the sixteenth century were not different from those seen in Mexico in present times. We present the humoral conceptions on which medical theory was based in those times, and the contributions made by Alonso Lopez de Hinojosos during his practice in the Hospital Real de San Josef de los Naturales, in Mexico City. Among them were the clinical distinction between gout and rheumatoid arthritis more than one hundred years before Sydenham, and the identification of arthritis and ocular involvement associated with a contagious disease more than three hundred years before Reiter. We conclude that the analysis of ancient medical traditions is an interesting and fruitful enterprise. PMID- 12089619 TI - ? PMID- 12089620 TI - [Physiopathology and biomechanics of hip osteoarthritis] AB - Several factors seem to play a relevant role in the pathogenesis of hip osteoarthritis. Among these, an altered biomechanic and neuromuscular integrity of the hip joint should be considered. This is a review of the recent international literature concerning the role of loads and strengths acting on the hip joint, in order to better understand the pathogenesis and the physiopathology of the hip osteoarthritis. The study of these factors might be important to prevent the development of the osteoarthritis and might suggest the conservative treatment. In particular, the role of the balance among the muscles working in maintaining the equilibrium of the acting strengths is matter of discussion. The articular and neuromuscular dysfunction might induce an altered load distribution in the hip, particularly on the articular cartilage surface, and seems to favour the development of hip osteoarthritis. PMID- 12089621 TI - [The subclinical involvement of the lung in rheumatoid arthritis: evaluation by high-resolution computed tomography] AB - Pulmonary involvement is one of the most frequent extra-articular manifestations of rheumatoid arthritis (RA) and represents a serious complication, being the second cause of death after infection. High-resolution computed tomography (HRCT), owing to its increased sensitivity and diagnostic accuracy respect to the conventional chest radiograph (CXR), allows to detect pulmonary abnormalities in RA patients more frequently than CXR. The aim of this study was to assess pulmonary involvement by HRCT in lifelong non-smoking RA patients without symptoms and clinical signs of pulmonary disease. Seventy-two patients (54 women and 18 men) with a mean age of 56.8+/-10.4 years (range, 40-77 years) and mean duration of disease of 6.9+/-4.7 years (range, 2-12 years) entered the study. 52/72 (72%) were positive for rheumatoid factor (> 20 UI/ml). Standard CXR and HRCT were carried out in each patient. CXR showed a mild interstitial fibrosis in 7 patients (9.7%), whereas HRCT demonstrated pulmonary abnormalities in an higher number of them (22/72 = 30.5%). The most frequent abnormal findings on HRCT were irregular pleural margins (13.8%) and septal/subpleural lines (18%), both compatible with pulmonary fibrosis. Ground-glass opacities were found in 8.3% of the patients. Pulmonary nodules (diameter, range 0,5-2 cm) predominantly located in the subpleural portions of the lung, were demonstrated in the same percentage (8.3%) of patients. Small airway involvement, represented by bronchiectasis/bronchioloectasis, was shown in 15.2% of patients. Subpleural cysts were present in two cases (2.8%). No patient had evidence of honeycombing on HRCT. In conclusion, HRCT is an accurate, non-invasive and safe method of diagnosing lung abnormalities in RA patients without signs and clinical symptoms of pulmonary disease. PMID- 12089622 TI - [Neurological involvement in systemic sclerosis. Evidence of a central Raynaud's phenomenon] PMID- 12089618 TI - [About "Multiple sclerosis and autoimmune diseases" ]. PMID- 12089623 TI - [Neonatal lupus: clinical features and risk of congenital cardiac heart block in newborns from mothers with anti Ro/SSA antibodies] AB - OBJECTIVE: To assess the prevalence of Congenital Heart Block (CHB) in newborns from anti Ro/SS-A antibodies positive mothers affected by connective tissue diseases (CTD) and to evaluate the prevalence of other manifestations of Neonatal Lupus (NL) and the electrocardiographic abnormalities. METHODS: A prospective study was conducted on 100 anti Ro/SS-A positive mothers that were followed before and during their 118 pregnancies (4 twin pregnancies and 18 second pregnancies). Counterimmunoelectroforesis (CIE) and immunoblot (IB) were used to test antibodies to extractable nuclear antigens (ENA). RESULTS: Only 2 cases of CHB (1.8%) were found among the 112 living newborns. In one case the mother with primary Sjogren's Syndrome (pSS) was anti Ro 60 and 52kD positive while in the other case the mother affected by undifferentiated connective tissue disease (UCTD) was anti Ro 60kD and anti La positive. No fetal death was due to CHB. There were no cutaneous rashes at birth while mild hepatic enzyme alterations were observed in 21 (68%) of the 31 tested newborns. In 22 healthy newborns an ECG have been registered and in 4 cases (18.2%) sinus bradycardia was found. During the follow up 7 suckling showed Cutaneous Neonatal Lupus. Moreover a six month girl developed Kawasaki Syndrome. CONCLUSIONS: The risk of delivering a child with CHB is 1.8% in anti Ro/SS-A positive mothers with CTD. This finding is extremely important in the preconceptional counseling of anti-Ro/SS-A positive women. Furthermore mild electrocardiographic abnormalities may be found in their healthy newborns. PMID- 12089624 TI - [Diff-Quik(R) staining method for detection and identification of monosodium urate and calcium pyrophosphate crystals in synovial fluids] AB - The aim of this study was to evaluate whether DQ could prove useful to identify monosodium urate (MSU) and calcium pyrophosphate dehydrate (CPPD) crystals on permanent mounted stained slides. To this end, we studied 27 synovial fluid (SF) samples obtained from the knees of patients with the pseudogout (n=21) and acute gouty arthritis (n=6). Wet analysis for crystal detection and identification was performed within one hour of joint aspiration. In addition, we studied 16 inflammatory synovial effusions obtained from patients with knee arthritis not induced by crystals. For each SF, DQ stained slides were analyzed by 2 experienced doctors in SF analysis. The observers were blinded to the type of crystal present in the SF. Each slide was analyzed by compensated polarized and transmitted light microscopy. SF was considered positive if intracellular and/or extracellular crystals were clearly identified. In addition, the observers were asked to identify the type of the crystals using compensated polarized light microscopy. Sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV) of the DQ staining method were determined. 51 true positive and 28 true negative specimens were correctly classified (39 CPPD samples, 12 MSU samples, and 28 samples of crystals-unrelated arthropathies). All MSU specimens were correctly diagnosed. PMID- 12089625 TI - [Chronic osteomyelitis: a new therapeutical idea] AB - We studied the case of a male patient aged 43 affected by post-traumatic chronic osteomyelitis with frequent relapses. Having supposed an insufficiency of the arterial and venous microcirculation in perilesional bone and soft tissue we decided for a therapy with iloprost and antibacterial drugs. After 15 months of treatment the patient hasn't showed any clinically evident relapsing episodes and we have not reported any side effects related to the therapy. PMID- 12089626 TI - [Compressive cervical myelopathy due to massive periodontoid calcium pyrophosphate crystal deposition] AB - A 77 year-old man suffering from psoriatic arthropathy presented with progressive myelopathy due to massive deposits of calcium pyrophosphate dihydrate crystals in peri-odontoid tissue. The magnetic resonance imaging and computer tomographic pictures of the involved site are shown and discussed. The clinical spectrum of crystal deposition disease involving the atlo-axial joint is briefly reviewed. PMID- 12089627 TI - [Sanitary costs of osteoarthritis] AB - Muscoloskeletal disorders are the first cause of disability and the second cause of permanent disablement in Italy. Osteoarthritis is the most frequent rheumatic disease and affects about 4 million Italians. In spite of that, data concerning social costs are lacking. On account of this lack we measured sanitary costs of 314 patients suffering from osteoarthritis. A retrospective, prevalence-based multicentric study was performed using a bottom-up approach. The study period was 12 months and referred to 1999. Eight percent of patients didn't take any drug for the treatment of osteoarthritis; NSAIDs were prescribed to 86.9% of patients, analgesics to 29.9%, chondroprotective drugs to 7.6%, and gastroprotective drugs to 36.9%. Total sanitary costs came to 455 euro / patient / year: 122 euro were spent on diagnostics, 293 euro on therapy and 40 euro on management of drug related gastropathy. Since the costs of anti-inflammatory drugs came to 30 euro we calculated iatrogenic cost factor of 2.3. Moreover, the study supplied interesting informations about prescriptive habits, which differ in Italy from international guidelines for the medical treatment of OA, about patient management, because of hospitalization, which by itself absorbs 1/3 of resources, and about physiotherapy, which costs twice as much as pharmacological therapy. At last, data analysis gave the cue for suggestions on changing patients' management. PMID- 12089628 TI - [Some historical notes on popliteal cysts] AB - Popliteal cysts, more appropriately those involving the gastrocnemio semimembranous bursa, are traditionally known as "Baker's cysts", since in 1877 the English surgeon William Morrant Baker published a paper on this topic ("On the formation of synovial cysts in the leg in connection with disease of the knee joint"). As a matter of facts, some authors described popliteal cysts before Baker. In particular, in 1856 a French physician, E.Foucher, performed a clinical and experimental study on 18 patients, focusing on some important aspects on the semeiology and pathophysiology of popliteal cysts. In 1861, Pietro Antivari, a physician from Friuli, a northeastern region of Italy, graduated at the University of Padova by producing an interesting paper on the popliteal cysts ("Dell'idrope delle borse mucose poplitee con tre osservazioni"). In the present historical note we report in full one out of three cases described by Pietro Antivari. PMID- 12089630 TI - Adverse effects of cell saver in patients undergoing ruptured abdominal aortic aneurysm repair. AB - A retrospective analysis of 56 patients undergoing ruptured abdominal aortic aneurysm (AAA) repair was performed to find out if cell saver had any impact on postoperative morbidity and mortality. All patients but one were male. The mean age was 68 +/- 8 years (35-85 years). Cell saver was used in 40 patients (CS group) and was not used in 16 patients (NCS group). We compared the incidences of respiratory, renal, and gastrointestinal complications; reoperation; transfusion requirement; length of hospital stay; and mortality between the groups. This study demonstrated that intraoperative cell saver usage significantly increased the incidence of respiratory complications and the need for blood and fresh frozen plasma transfusion, and prolonged the hospital stay in patients with ruptured AAA, but did not have any impact on mortality. Postoperative complications were more prominent in patients who received >3000 mL cell saver blood. PMID- 12089629 TI - Biochemical alterations in cerebrospinal fluid during thoracoabdominal aortic cross-clamping in dogs. AB - Spinal cord damage during and after thoracoabdominal aortic cross-clamping continues to be a major problem. Somatosensory and motor evoked potentials have been used to monitor spinal cord function but their value for predicting paraplegia has been controversial. The aim of this study was to measure biochemical markers in the cerebrospinal fluid (CSF) and correlate changes with spinal cord ischemia. Since neural tissue utilizes only glucose as substrate for its metabolism and energy supply, we measured changes of metabolites of anaerobe glycolysis. In a canine model in which general anesthesia was used, the thoracoabdominal aorta was cross-clamped proximally and distally for 60 min. Hemodynamic parameters, blood gases, and glucose level were monitored continuously. Blood and CSF sampling were performed at baseline, at 15, 30, and 55 min during cross-clamping, and at 5 and 15 min after aortic declamping. Levels of lactate (1.7 +/- 0.1 to 3.2 +/- 0.3 mmol/L), pCO2 (43 +/- 2 to 35 +/- 1.6 mmHg), and neuron-specific enolase (NSE) (5.17 +/- 0.5 to 13.0 +/- 3.5 mg/L) in CSF showed significant changes (p < 0.05) during clamping and reperfusion. Changes in CSF lactate and NSE levels correlate with the duration of spinal cord ischemia. These markers of ischemic metabolism appear suitable to monitor the degree of spinal cord ischemia during thoracoabdominal cross-clamping and may be useful to predict the efficacy of preventive methods. PMID- 12089631 TI - Splenic artery aneurysms: two decades experience at Mayo clinic. AB - Although rare, splenic artery aneurysms (SAAs) have a definite risk of rupture. The optimal management of these aneurysms remains elusive. A retrospective chart review of all patients treated at our institutions with the diagnosis of SAA from January 1980 until December 1998 was undertaken. Follow-up was obtained via chart review and by direct phone contact of the patient or relative. No specific protocol was followed for management. From analysis of the patient data we concluded that although SAAs may rupture, not all intact aneurysms need intervention. Calcification does not appear to protect against rupture, although beta-blockade may be protective. Growth rates of SAA are slow and growth is infrequent. Selective management of SAAs is safe. Open ligation or transcatheter embolization should be considered for symptomatic aneurysms, for aneurysms > or = 2 cm in size, or for any SAA in women of childbearing years. PMID- 12089632 TI - Hand-assisted laparoscopic autoaugmentation gastrocystoplasty. AB - BACKGROUND: Autoaugmentation gastrocystoplasty has been previously performed successfully. This set of experiments was conducted to determine the feasibility of performing autoaugmentation gastrocystoplasty laparoscopically. METHODS: Hand assisted laparoscopic autoaugmentation gastrocystoplasty was performed on 15 mongrel dogs. The surgery was carried out with two 10-mm trocars and a 6-cm Pfannenstiel incision. The gastric wedge, supplied by the right gastroepiploic artery, was resected with two applications of an endoscopic gastrointestinal anastomosis (GIA) stapler. The pedicle was demucosalized, and the anastomosis to the bladder was completed through the Pfannenstiel incision. RESULTS: All of the dogs were successfully treated laparoscopically and were eating at 48 h. There was no evidence of anastomotic leak dehiscence at the gastric resection staple line. CONCLUSION: Hand-assisted laparoscopic autoaugmentation gastrocystoplasty can be performed successfully in dogs. This operation may offer a superior alternative to standard bladder autoaugmentation procedures in children suffering from congenital bladder disorders. PMID- 12089633 TI - Concomitant laparoscopic splenectomy and cholecystectomy as an effective and minimally invasive treatment of pyruvate kinase deficiency with gallstones. AB - A 4-year-old girl with pyruvate kinase deficiency underwent partial splenic embolization initially. However, even after this procedure, she still had to be transfused every 2 months and then every month. At 5 years of age, she was admitted to our hospital to undergo splenectomy. She underwent laparoscopic splenectomy and concomitant cholecystectomy for gallstones. The hemogram recovered to the normal range after surgery, and her postoperative course was uneventful. Considering the absence of morbidity, the short hospitalization, the quick return to normal activity, the good cosmetic result, and the improved clinical and hematologic results, we consider that simultaneous laparoscopic splenectomy and cholecystectomy is safe and effective for the management of hemolytic anemia resulting from pyruvatre kinase deficiency and associated with cholelithiasis. PMID- 12089634 TI - Repair of diaphragmatic rupture by laparoscopic implantation of a polytetrafluoroethylene patch. AB - Diaphragmatic hernias after blunt traumatic damage are serious complications, and diagnosis often might be delayed. In most cases, early symptoms are missed, but in the further posttraumatic period, patients experience recurrence of pulmonary dysfunction or intestinal symptoms such as obstruction, nausea, and pain. Most of these defects are diagnosed by laparotomy performed to investigate other major abdominal lesions. These diaphragmatic ruptures are managed by suturing using a thoracic or abdominal approach. An original diaphragmatic repair technique using a patch is presented. A 40-year-old woman was admitted to our clinic because of chronic abdominal pain. Chest x-ray and computed tomography scan showed a migration of the large intestine into the left hemithorax. A large diaphragmatic hernia was diagnosed and repaired laparoscopically using a patch. In the reported patient, laparoscopic suture of a diaphragmatic hernia using a polytetrafluoroethylene (PTFE) (GORETEX) patch proved to be safe, successful, elegant, and uneventful. PMID- 12089635 TI - Promotion of safety helmets for child bicyclists: 2002 update. AB - PURPOSE: Current research continues to show that bicycle helmets prevent serious injury and death in cyclists of all ages. Children are at special risk for head injury. The purpose of this review was to update information on the use and protective effect of bicycle helmets for child cyclists. CONCLUSIONS: Legislation, combined with community education and helmet promotion campaigns, remains the most cost-effective approach for increasing helmet use. Nineteen states and the District of Columbia have passed helmet legislation. Modest gains in helmet-wearing rates have been documented, with the greatest gains in young children, compared with adolescents. IMPLICATIONS: Nurses remain in strategic positions to encourage helmet use by educating children and parents. Nurses can participate in community-based education and advocacy programs. Nurse researchers can carry out descriptive studies that increase community awareness of bicycle injuries and examine factors such as parental rules and adolescent risk-taking. PMID- 12089636 TI - An integrative review of patients' perceptions regarding telehealth used in their health care. AB - The rapid changes in technology and communication have had a great impact on the delivery of health care in the United States. Such changes have spurred the growth of an arena in health care known as telehealth. An important medium in telehealth care is the use of video. This paper examines the research that has been undertaken in determining patients perceptions of the use of a video component in their health care. Ten studies examined used real-time, interactive video to deliver care, while one evaluated still images. The results of the integrative review tentatively suggest that patients are amenable to the use of video in telehealth interventions. Limitations of the studies reviewed also are discussed. PMID- 12089637 TI - Improving the quality of pain assessment through research utilization. PMID- 12089638 TI - Unit-based specialty vascular transitional home care program: an example of evidence-based nursing practice. PMID- 12089639 TI - Using a FOCUS-PDCA quality improvement model for applying the severe traumatic brain injury guidelines to practice: process and outcomes. PMID- 12089640 TI - Evidence-based development of a hospital-based heart failure center. PMID- 12089641 TI - Neonatal pain management with oral sucrose. PMID- 12089642 TI - [Idiopathic interstitial pneumonia - why again a new consensus classification?]. PMID- 12089643 TI - [The song of the humming birds: A series an rare pulmonary diseases--start of a new subject column]. PMID- 12089644 TI - [Positive pressure ventilation in Cheyne-Stokes respiration]. AB - Cheyne-Stokes respiration (CSR) is found in patients with chronic left ventricular failure and associated with a reduced prognosis. Continuous positive airway pressure (CPAP) improves the survival rate. In this retrospective study we report on the effect of different positive pressure ventilation modes in CSR. The observation period lasted from 1995 - 1999. Inclusion criteria was CSR with a respiratory disturbance index > 10/h whereas > 50 % of the events had to be central and/or mixed. In each patient a 4 week lasting intervention with each of CPAP, Bilevel CPAP in spontan mode (i.e. BiPAP S or BiPAP ST) and BiPAP in controlled mode (i.e. BiPAP T) was performed. Responder were defined by subjective and objective criteria. In total 41 males were included. Responder were distributed as follows: CPAP: n = 13 patients (31.7 %), BiPAP S/ST: n = 9 patients (22.0 %) and BiPAP T: n = 12 patients (29.3 %). In total 7 from 41 patients (17.1 %) rejected a long-term treatment with positive pressure ventilation. The majority of patients with CSR responded to CPAP and BiPAP S/ST mode. Compared to the other responder groups the best quality was reached with BiPAP T. However the latter was performed only by 29 % of the population. PMID- 12089645 TI - [Prevalence of obstructive airway disease in middle-age adults. Cross sectional study in three different occupations]. AB - Medical management of COPD begins with an awareness of risk factors and identification of risk patients. To identify this patients, 179 healthy volunteers with no known obstructive airway disease (81 women, 98 men, 40,5 +/- 11,2 years) and three different occupations (43 factory workers, 33 community workers, mainly working outside and 113 office employees) participated in the cross sectional spirometry study. Demographic data concerning age, height and body weight were comparable. In the first two groups were more men and more smokers. After adjustment to sex, age and height smokers revealed significant lower data for FEV 1 (3,16 l vs. 3,44 l) and FEV 1 /IVC (74,5 % vs. 77,6 %) in the factory workers. Comparable data exists for smokers in the community workers for FEV 1 (3,21 l vs. 3,59 l) and FEV 1 /IVC (74,8 % vs. 79,6 %), but not for office employees (FEV 1 3,51 l vs. 3,54 l; FEV 1 /IVC 76,1 % vs. 78,4 %). Airway obstruction, defined by FEV 1 /IVC below 70 % could be identified in thirteen of 57 smokers (22,8 %) and in twelve of 122 non-smokers (9,8 %) or in 25 of all 179 volunteers (14 %). The influence of smoking was significant (P = 0,0123). As a consequence, first there is a need for an increased awareness and routine spirometry in ad-risk patients; second, education and smoking cessation programs have to be initiated. PMID- 12089646 TI - [Continuous acoustic monitoring of nocturnal bronchial obstructions]. AB - Patients with bronchial asthma often have respiratory problems in sleep. The effects of bronchial obstructions while sleeping have been analysed in some studies. For an exact assessment the sleep itself must not be disturbed by the method. The continuous acoustic lung sound detection is such a method. It helps to assess the circadian rhythm during antiobstructive therapy which may lead to a better sleep quality and daytime fitness. PMID- 12089647 TI - [European Community Respiratory Health Survey in Adults (ECRHS)]. AB - The European Community Respiratory Health Survey (ECRHS) was the first study to assess the geographical variation in asthma, allergy, and allergic sensitization in adults using the same instruments and definitions. The database of the ECRHS includes information from approximately 140 000 individuals aged 20 - 44 years from 22 countries. The aim of this review is to summarize the results of the ECRHS and to present the specific contribution of the German centers in Hamburg and Erfurt. The prevalence ranged from 2.0 - 11.9 % for asthma, 9.5 - 40.9 % for allergic rhinitis, 4.0 - 32.0 % for wheeze, 3.4 - 27.9 % for bronchial hyperreactivity, and 16.2 - 44.5 % for allergic sensitisation against common aeroallergens. Although the prevalence of these atopic disorders were found to be consistently higher for the Hamburg center compared to the Erfurt center, strong regional differences in the prevalences were also found within several other European countries. Overall Europe, the lowest prevalences were seen in the Eastern and Middle European countries with the center Erfurt, followed by the Mediterranean region. The highest prevalences were reported for all English speaking centers. Strong geographic variation was reported for medication for asthma. Asthma seems to be undertreated in several countries. Environmental exposures and in particular indoor factors, and exposures at the workplace are playing a major role for asthma in adulthood. Furthermore, protective effects on atopy were found for exposures to pets (dogs) and a large number of siblings in early childhood. In conclusion, the ECRHS has shown that the prevalence of asthma varies widely. The fact that the geographical pattern is consistent with the distribution of atopy and bronchial responsiveness supports the conclusion that the geographical variations in the prevalence of asthma are true and likely due to environmental factors. PMID- 12089648 TI - ["When should a substance be designated as sensitizing for the skin ("sh") or for the airways ("sa")"?"]. PMID- 12089649 TI - [Pulmonary lymphangioleiomyomatosis]. PMID- 12089650 TI - [Pharmacological treatment of epilepsies: progress during the last 10 years]. AB - The pharmacological treatment of epilepsies represents an important domain of neurological therapeutical strategies and is not only important for specialized epilepsy centres, but for all hospital -and office-based neurologists. The choice of the right anticonvulsive drug is not always simple since the development of new drugs during the last 10 years offers a broad range of possibilities. Therefore, pharmacological treatment has become more sophisticated than it was with relatively few anticonvulsive drugs before. The new drugs are better tolerated and have less side effects, but more knowledge is required about the specific aspects of each drug. Pharmacology, pharmacocinetics, indication, dosage and side effects of the substances will be summarized and discussed. PMID- 12089651 TI - [Current use of ECT in the treatment of depressive disorders]. AB - If used with correct indication, electroconvulsive therapy (ECT) is a very effective and safe treatment for some forms of major depression, provided due consideration is given to possibly concomitant somatic conditions and that the cardiovascular reactions during and after the ECT treatments are managed carefully. Despite this, ECT remains controversial, especially in Germany. This article provides an overview of the current state of the art in ECT: appropriate indications, best practice in conducting ECT, side effects and recent developments in the use of ECT as maintenance therapy. This, it is hoped, will contribute to a more factually grounded discussion of the use of ECT. PMID- 12089652 TI - [Kurt Goldstein's understanding of amnesic aphasia and its underlying disorder - an early model of the pensee operatoire of the French psychosomatic school?]. AB - Kurt Goldstein's understanding of amnesic aphasia in some regards anticipated the model of the pensee operatoire, a concept developed during the 60's and 70's by the French psychoanalytical school of psychosomatics. Goldstein interpreted amnesic aphasia within the framework of a "basic disorder". Closely following the philosopher Ernst Cassirer, Goldstein described amnesic aphasia as an expression of a general alteration following localized or generalised brain damage. Due to various historical events (world war, fascism, the holocaust) as well as developments during the 20(th) century (dominance of the English language in many areas of science), these connections were forgotten or were no longer recognised as such. Without wanting to determine the extent to which the concept of pensee operatoire possesses validity, one can interpret Goldstein's reflections on aphasia as a heretofore unreceived preliminary model of the psychosomatic concept of the French School. PMID- 12089653 TI - [Current standing of forensic psychotherapy in Germany]. AB - Although treatment of inmates already has got a long tradition in Germany, forensic psychotherapy is - especially outside the so-called German "Massregelvollzug" - only at its beginnings, and consistent theories about delinquency and treatment programmes have not yet been established. While international studies show high prevalence of mental disorders in prisons (up to 60 - 70 %), in Germany even basic epidemiological data about the prevalence of mental disorders among inmates is still missing. Thus, the need for psychotherapy can only be roughly estimated. Ethical implications regarding the self-image, an ambivalent assignment for treatment (given by society) and a recent tightening of German law create a difficult framework for forensic psychotherapy, complicating the establishment as well as the evaluation of new treatment programmes, especially since many people still doubt the efficiency of forensic psychotherapy for delinquents despite convincing counter-evidence. It seems obvious that in Germany much research has to be done in the fields of epidemiology as well as in forensic psychotherapy in order to gain basic data and to be then able to provide effective psychotherapeutic treatment facilities aiming at the needs of mentally disturbed offenders. PMID- 12089655 TI - Epidemiologic and molecular trends of "Norwalk-like viruses" associated with outbreaks of gastroenteritis in the United States. AB - Between July 1997 and June 2000, fecal specimens from 284 outbreaks of nonbacterial gastroenteritis were submitted to the Centers for Disease Control and Prevention for testing for "Norwalk-like viruses" (NLVs). Specimens were examined by reverse-transcription polymerase chain reaction and direct electron microscopy for the presence of NLVs. Adequate descriptive data were available from 233 of the outbreaks, and, of these, 217 (93%) were positive for NLVs. Restaurants and events with catered food were the most common settings, and contaminated food was the most common mode of transmission. Genogroup II (GII) strains were the predominant type (73%), with genogroup I strains causing 26% of all NLV-positive outbreaks. Certain GII clusters (GII/1,4,j) were more commonly associated with outbreaks in nursing home settings than with outbreaks in other settings. Strain diversity was great: one potential new sequence cluster was implicated in multiple outbreaks, and strains belonging to a tentative new genogroup were identified. PMID- 12089654 TI - Autosomal dominant stapes ankylosis with broad thumbs and toes, hyperopia, and skeletal anomalies is caused by heterozygous nonsense and frameshift mutations in NOG, the gene encoding noggin. AB - Although fixation of the stapes is usually progressive and secondary to otosclerosis, it may present congenitally, with other skeletal manifestations, as an autosomal dominant syndrome-such as proximal symphalangism (SYM1) or multiple synostoses syndrome (SYNS1), both of which are caused by mutations in NOG, the gene encoding noggin. We describe a family that was ascertained to have nonsyndromic otosclerosis but was subsequently found to have a congenital stapes ankylosis syndrome that included hyperopia, a hemicylindrical nose, broad thumbs and great toes, and other minor skeletal anomalies but lacked symphalangism. A heterozygous nonsense NOG mutation-c.328C-->T (Q110X), predicted to truncate the latter half of the protein-was identified, and a heterozygous insertion in NOG c.252-253insC, in which the frameshift is predicted to result in 96 novel amino acids before premature truncation-was identified in a previously described second family with a similar phenotype. In contrast to most NOG mutations that have been reported in kindreds with SYM1 and SYNS1, the mutations observed in these families with stapes ankylosis without symphalangism are predicted to disrupt the cysteine-rich C-terminal domain. These clinical and molecular findings suggest that (1) a broader range of conductive hearing-loss phenotypes are associated with NOG mutations than had previously been recognized, (2) patients with sporadic or familial nonsyndromic otosclerosis should be evaluated for mild features of this syndrome, and (3) NOG alterations should be considered in conductive hearing loss with subtle clinical and skeletal features, even in the absence of symphalangism. PMID- 12089656 TI - Differential expression of proinflammatory cytokine genes in vivo in response to pathogenic and nonpathogenic pneumovirus infections. AB - Pneumonia virus of mice (PVM; Paramyxoviridae, subfamily Pneumovirinae) is an important pathogen for the study of physiologically relevant acute inflammatory responses in rodent hosts. In contrast to the severe symptomatology observed in response to infection with PVM strain J3666, infection with strain 15 resulted in few clinical symptoms, limited cellular inflammatory response, and no production of macrophage inflammatory protein-1alpha or monocyte chemoattractant peptide (MCP)-1. Microarray analysis of transcripts from lung tissue indicates that PVM J3666 infection promotes up-regulation of specific proinflammatory genes, most notably interferon (IFN)-1beta, IFN response genes, and chemokines MCP-1, MCP-3, RANTES (regulated on activation, normally T cell-expressed and secreted), and eotaxin. Of these, only RANTES expression increased in response to infection with strain 15, with no increased expression of IFN or IFN response genes, despite ongoing viral replication. These results suggest that pneumovirus replication alone is insufficient to promote antiviral inflammation and that evaluation of the more divergent strain-specific pneumovirus proteins may provide some intriguing leads toward the molecular basis of this differential response. PMID- 12089658 TI - Incidence of and risk factors for severe hepatotoxicity associated with antiretroviral combination therapy. AB - This retrospective cohort study investigated whether particular antiretroviral agents are associated with a higher risk for developing grade 4 liver enzyme elevations (LEEs) in patients with human immunodeficiency virus (HIV) type 1 infection who are starting to receive highly active antiretroviral therapy (HAART). Grade 4 LEE was defined as aminotransferase levels >10 times the upper limit of normal and >200 U above baseline levels. A multivariate Cox model was used to identify risk factors. The incidence of LEE was 6.3%. No patients died of LEE consequences. Risk factors were higher baseline alanine aminotransferase levels, chronic hepatitis B or C virus infection, antiretroviral therapy-naive patients undergoing their first HAART regimen, recent start of a regimen of nevirapine or high-dose ritonavir, and female sex. In hepatitis B virus (HBV) coinfected patients, discontinuing lamivudine (3TC) use was a risk factor. In 97% of cases, >or=1 risk factor was present. In HBV-coinfected patients using 3TC, continued use of 3TC should be considered, even if 3TC-resistant HIV strains develop. PMID- 12089657 TI - Dynamics of CD4 and CD8 T cell responses to cytomegalovirus in healthy human donors. AB - To study the dynamics of cytomegalovirus (CMV) immunity in healthy immunocompetent hosts, interferon-gamma-producing CD4 and CD8 T cell responses in the presence or absence of CMV antigens were measured from 15 CMV-seropositive donors and 13 CMV-seronegative donors. Cytokine responses in the absence of antigen were significantly higher in CMV-seropositive donors. Also, a disproportionate number of CD69(+) cells isolated ex vivo from CMV-seropositive donors were specific for CMV, suggesting recent reactivation in vivo. To examine changes in cellular responses over time, 10 seropositive donors were tested over a 6-month period. About half of the donors showed significant variability over time, but staphylococcal enterotoxin B responses remained relatively constant. These findings suggest that CMV can present a considerable and recurrent burden to the human immune system. By understanding the normal dynamics of CMV responses over time, it may be possible to better identify aberrant responses to CMV in immunocompromised hosts. PMID- 12089659 TI - Platelet-binding domains in 2 fibrinogen-binding proteins of Staphylococcus aureus identified by phage display. AB - The adherence of microorganisms to platelets previously immobilized on the subendocardium in nonbacterial thrombotic endocarditis is considered an important pathogenic step in Staphylococcus aureus endocarditis. To identify and characterize bacterial factors involved in the adherence to platelets, a phage display library of S. aureus was generated by use of the phagemid pG8H6. The library was affinity panned against purified immobilized platelets. After a second panning against platelets, a significant increase in the number of eluted phagemid particles was observed; 27% of 88 randomly isolated clones expressed overlapping deduced amino acid sequences with high similarity to the C-terminal domain of the S. aureus coagulase. In addition, 22% of the clones expressed the N terminal domain of the fibrinogen-binding protein Efb. The surface-associated fraction of the C-terminal domain of coagulase or the N-terminal domain of Efb may be involved in bacterial adherence to immobilized platelets, and fibrinogen may act as a bridging molecule in that interaction. PMID- 12089660 TI - Genotype-specific carriage of Neisseria meningitidis in Georgia counties with hyper- and hyposporadic rates of meningococcal disease. AB - Carriage of Neisseria meningitidis in a Georgia county with hypersporadic incidence of meningococcal disease ("hypersporadic county") and in a county with no cases of meningococcal disease was determined by a cross-sectional pharyngeal culture study of high school students. Among 2730 students from whom culture samples were obtained, meningococcal carriage was 7.7% (140/1818) in the hypersporadic county and 6.1% (56/912) in the comparison county. Carriage rates by serogroup and genetic type (i.e., electrophoretic type [ET]) did not differ significantly between counties, but apartment or mobile home residency was a risk factor for carriage in the hypersporadic county. Although most cases of meningococcal disease in the hypersporadic county were caused by members of the serogroup C ET-37 clonal group, no ET-37 meningococcal isolates were recovered from carriers in this county. However, 38% of all meningococcal isolates recovered from carriers in both counties were members of the serogroup Y ET-508 clonal group, an emerging cause of meningococcal disease in Georgia and throughout the United States during 1996-2001. Shifts in carriage and transmission of meningococcal strains with different pathogenic potential are important determinants of meningococcal disease incidence. PMID- 12089661 TI - Evidence of extensive interspecies transfer of integron-mediated antimicrobial resistance genes among multidrug-resistant Enterobacteriaceae in a clinical setting. AB - Multidrug resistance in gram-negative bacteria appears to be primarily the result of the acquisition of resistance genes by horizontal transfer. To what extent horizontal transfer may be responsible for the emergence of multidrug resistance in a clinical setting, however, has rarely been investigated. Therefore, the integron contents of isolates collected during a nosocomial outbreak of genotypically unrelated multidrug-resistant Enterobacteriaceae were characterized. The integron was chosen as a marker of transfer because of its association with multiresistance. Some genotypically identical isolates harbored different integrons. Grouping patients carrying the same integron yielded 6 epidemiologically linked clusters, with each cluster representing a different integron. Several patients carried multiple species harboring the same integron. Conjugation experiments with these strains resulted in the transfer of complete resistance patterns at high frequencies (10(-2) to 10(-4)). These findings provide strong evidence that the horizontal transfer of resistance genes contributed largely to the emergence of multidrug-resistant Enterobacteriaceae in this clinical setting. PMID- 12089662 TI - Escherichia coli O157:H7 infections: discordance between filterable fecal shiga toxin and disease outcome. AB - Shiga toxin (Stx)-producing Escherichia coli O157:H7 are the most common cause of hemolytic uremic syndrome (HUS). We detected free fecal Stx in 48%, 40%, and 17% of infected children with uncomplicated diarrhea, children who subsequently developed HUS, and children with HUS, respectively. Vero cell assay detected Stx more frequently than did a commercial Stx enzyme immunoassay. In children's stool samples obtained on or before day 4 of illness, each 10-fold decrease in titer was, paradoxically, associated with 3.8-fold increased odds of developing HUS (P=.03; 95% confidence interval, 0.77-19.7). The fecal Stx type did not correlate with the Stx expressed by bacteria grown in vitro and was not related to bacterial titer in the studied samples. These data suggest that therapeutic and diagnostic strategies directed toward binding or identifying intraintestinal fecal Stx may have limited success. PMID- 12089663 TI - Generation and characterization of a protective monoclonal antibody to Pseudomonas aeruginosa PcrV. AB - Pseudomonas aeruginosa is a gram-negative pathogen causing life-threatening infections. Lung injury and the development of sepsis depend largely on the expression of type III secretion system (TTSS) virulence. TTSS functions as a molecular syringe to deliver toxins directly to the cytosol of cells, inhibit innate immune mechanisms, and prevent bacterial clearance. Polyclonal antibodies that bind to PcrV of P. aeruginosa inhibit the delivery of type III toxins and enhance the clearance of bacteria during acute lung infections. PcrV is a homologue of LcrV, a protective antigen in the Yersinia TTSS and an integral component of TTSS. In this study, a murine monoclonal antibody (MAb) to PcrV was generated: MAb 166, which is protective against P. aeruginosa when coinstilled with the bacterial inoculum or intraperitoneally transferred to mice. Fab fragments from MAb 166 prevent sepsis and death. The epitope bound by MAb 166 was mapped to the carboxyl-terminus of PcrV. PMID- 12089664 TI - Genomic deletions suggest a phylogeny for the Mycobacterium tuberculosis complex. AB - To better understand the evolution of the Mycobacterium tuberculosis complex, subspecies were tested for large sequence polymorphisms. Samples with greater numbers of deletions, without exception, were missing all the same regions that were deleted from samples with lesser numbers of deletions. Principal genetic groups based on single-nucleotide polymorphisms were restricted to one of the deletion-based groups, and isolates that shared genotypes based on molecular epidemiological markers were assigned almost exclusively to the same deletion type. The data provide compelling evidence that human tuberculosis did not originate from the present-day bovine form. Genomic deletions present themselves as an attractive modality to study the evolution of the M. tuberculosis complex. PMID- 12089665 TI - An outbreak of neonatal deaths in Brazil associated with contaminated intravenous fluids. AB - A nursery outbreak of fever and clinical sepsis resulted in the deaths of 36 neonates in Roraima, Brazil. To determine the cause, epidemiologic studies were performed, along with culture and endotoxin analysis of intravenous (iv) fluids. Affected neonates were more likely to have lower birth weight (2.1 vs. 3.2 kg; P<.01), lower APGAR (activity, pulse, grimace, appearance, and respiration) score at 1 (7 vs. 8; P=.1) or 5 min (8 vs. 9; P=.03), lower gestational age (32 vs. 39 weeks; P=.001), or to receive iv medications (20/20 vs. 2/40; P<.0001). Fever occurred only after iv medication administration. Although culture results of unopened iv medications were negative, endotoxin levels of glucose and distilled water for injection were elevated (3.3 and 1.2 U/mL, respectively). Endotoxin contaminated iv medications were distributed nationally and may have caused other outbreaks of unexplained death. These results highlight the importance of monitoring both pharmaceutical quality and postmarketing surveillance for adverse events. PMID- 12089666 TI - Type 1 T helper cells specific for Candida albicans antigens in peripheral blood and vaginal mucosa of women with recurrent vaginal candidiasis. AB - The cytokine profile of circulating and vaginal T cells specific for immunodominant mannoprotein antigens of Candida albicans was analyzed in patients with recurrent vaginal candidiasis (RVC). Peripheral blood mononuclear cells (PBMC) from patients with RVC proliferated more than those from healthy subjects and expressed higher type 1:type 2 T helper cell cytokine ratios in response to C. albicans stimulation. A higher number of C. albicans-specific T cells was generated in PBMC from patients with RVC than in PBMC from healthy donors. C. albicans-specific T cell clones from patients with RVC produced higher levels of interferon (IFN)-gamma and lower levels of interleukin (IL)-4 than clones from control women. More important, a higher proportion of C. albicans-specific T cell clones was generated from lesional mucosa of patients with RVC than from normal mucosa, all of which produced IFN-gamma but not IL-4. These findings provide direct evidence that RVC is characterized by a highly polarized local and circulating type 1 T helper cell-like response against C. albicans antigens. PMID- 12089667 TI - Cryptosporidiosis stimulates an inflammatory intestinal response in malnourished Haitian children. AB - The mechanisms by which Cryptosporidium parvum cause persistent diarrhea and increased morbidity and mortality are poorly understood. Three groups of Haitian children <18 months old were studied: case patients, children with diarrhea not due to Cryptosporidium, and healthy control subjects. Compared with both control groups, children with acute cryptosporidiosis were more malnourished (including measures of stunting [P=.03] and general malnutrition [P=.01]), vitamin A deficient (P=.04), and less often breast-fed (P=.04). Markers of a proinflammatory immune response, interleukin (IL)-8 and tumor necrosis factor alpha receptor I, were significantly elevated in the case population (P=.02 and P<.01, respectively), as was fecal lactoferrin (P=.01) and the T helper (Th)-2 cytokine IL-13 (P=.03). The counterregulatory cytokine IL-10 was exclusively elevated in the case population (P<.01). A Th1 cytokine response to infection was not detected. This triple cohort study demonstrates that malnourished children with acute cryptosporidiosis mount inflammatory, Th-2, and counterregulatory intestinal immune responses. PMID- 12089668 TI - Younger age at vaccination may increase risk of varicella vaccine failure. AB - To determine vaccine effectiveness (VE), a varicella outbreak in a highly vaccinated day-care center (DCC) population in Pennsylvania was investigated. In Pennsylvania, proof of immunity is required for children >or=12 months old for DCC enrollment. Questionnaires were administered to parents of children who had attended the DCC continuously during the study period (1 November 1999-9 April 2000) to determine history of varicella disease or vaccination and for information about any recent rash illnesses. VE was calculated for children >or=12 months old without a history of varicella. There were 41 cases of varicella among 131 attendees, with 14 cases (34%) among vaccinated children. VE was 79% against all varicella and 95% against moderate or severe varicella. Vaccination at <14 months was associated with an increased risk of breakthrough disease (relative risk, 3.0; 95% confidence interval, 0.9-9.9). Despite varicella vaccination coverage of 80%, a sizeable outbreak occurred. Early age at vaccination may increase the risk of vaccine failure. PMID- 12089669 TI - Human leukocyte antigen class II alleles may contribute to the severity of hepatitis C virus-related liver disease. AB - Whether the host's immune response genes influence the severity of hepatitis C virus (HCV) liver disease is controversial. Human leukocyte antigen (HLA) class II alleles were analyzed in 233 HCV RNA-positive patients with chronic active hepatitis (197 patients with Knodell index of fibrosis F0-F3 and 36 patients with index of F4). The 2 groups did not differ by sex, duration of infection, mode of contamination, alcohol consumption, or HCV genotype. Patients with cirrhosis were older than those without (56+/-12 vs. 46+/-14 years; P<10-4) and had a lower DRB1*11 allele frequency (5.6% vs. 14.5%; P=.037), whereas DRB1*03 and DQB1*0201 frequencies appeared to be higher (DRB1*03, 18.1% vs. 9.6%; DQB1*0201, 37.5% vs. 23.4%; P=.04, corrected P value is not significant). Mean index of fibrosis was higher in DR3-positive than in DR11-positive patients (2.14 vs. 1.58; P=.05). By multivariate analysis, cirrhosis was associated with male sex and age >50 years. HLA class II alleles may weakly contribute to the severity of HCV liver disease. Of persons infected with HCV, only 15%-20% spontaneously clear the virus, and the rest become chronically infected. PMID- 12089670 TI - Prognostic value of quantitative Kaposi sarcoma--associated herpesvirus load in posttransplantation Kaposi sarcoma. AB - Organ transplant recipients have a higher risk of Kaposi sarcoma (KS). A quantitative real-time polymerase chain reaction assay was developed to evaluate KS-associated herpesvirus (KSHV) as a prognostic tool in transplant recipients with KS. Forty-three patients who developed KS after transplantation were included in a cross-sectional study to correlate virus load with transplantation or KS parameters. Seventeen patients (40%) had KSHV viremia (>100 copies/microg of DNA; median, 6067 copies/microg of DNA). Factors associated with these levels of viremia by univariate analysis were progression of KS (P=.00002), time from KS diagnosis (P=.0007), actual stage of KS (P=.006), initial stage of KS (P=.22), graft loss (P=.013), and time from transplantation (P=.0246). Disease progression remained associated with KSHV viremia in a multivariate analysis (P=.01). Thus, quantification of KSHV load in peripheral blood mononuclear cells could represent a useful tool for monitoring transplant recipients with KS. PMID- 12089671 TI - Cytomegalovirus glycoprotein B groups in human immunodeficiency virus-infected patients with incident retinitis. AB - Cytomegalovirus (CMV) strains may be categorized into 4 different groups on the basis of glycoprotein B (gB) genotype. gB genotypes in CMV polymerase chain reaction (PCR)-positive samples from case patients who were diagnosed with retinitis during prospective follow-up were compared with genotypes in CMV PCR positive samples from an equal number of retinitis-free matched control subjects. All patients were infected with human immunodeficiency virus (HIV) and CMV. Control subjects and their plasma samples were matched with case patients according to baseline CD4(+) T cell count, transfusion history, HIV risk factor, and follow-up time. CMV DNA was genotyped by restriction-enzyme digestion. Eighteen patients met our case definition. Approximately one-third of patients had gB genotype 1, and half had gB genotype 2. These rates were similar among case patients and control subjects and were similar by risk group. The CMV gB2 genotype is not a major determinant of retinitis pathogenicity but appears to be highly prevalent among HIV-infected patients. PMID- 12089673 TI - Conjugate vaccines against group B Streptococcus types IV and VII. AB - Although rarely encountered, group B Streptococcus (GBS) types IV and VII have been isolated from infants and adults with invasive disease. This study was designed to determine the immunogenicity and efficacy in animals of conjugate vaccines prepared with GBS types IV and VII capsular polysaccharide (CPS). Despite the striking similarities in structure of these 2 carbohydrate antigens, high-titer rabbit antiserum to each conjugate vaccine was serotype specific. Active vaccination of female mice with the conjugate vaccines induced type specific IgG and resulted in survival of >90% of newborn pups challenged with viable GBS of homologous serotype. If needed, types IV and VII CPS conjugate vaccines of the design described can be added to the formulation of a multivalent GBS vaccine. PMID- 12089672 TI - Emergence of a novel penicillin-nonsusceptible, invasive serotype 35B clone of Streptococcus pneumoniae within the United States. AB - Monitoring antibiotic-resistant pneumococcal strains not covered by the 7-valent conjugate vaccine is an important priority. The Centers for Disease Control and Prevention's Active Bacterial Core Surveillance identified 68 invasive penicillin nonsusceptible serotype 35B (PN35B) isolates recovered from 1995 to 2001 from patients residing in the states of California, Colorado, Connecticut, Georgia, Maryland, Minnesota, New York, Oregon, Tennessee, and Texas. Nonsusceptible isolates accounted for 69% of all serotype 35B isolates recovered during this time. Twelve (18%) of the 68 PN35B isolates recovered since 1995 were obtained from pediatric patients. These 68 isolates exhibited penicillin MICs of 0.25-2 microg/mL and reduced susceptibility to cefotaxime. Representative PN35B isolates exhibited a common chromosomal macrorestriction profile and identical penicillin binding-protein gene restriction profiles characteristic of penicillin-resistant strains, and they shared a unique 7-locus sequence type that included 3 new alleles. The mosaic pbp2b and divergent ddl sequences were suggestive of interspecies recombination at the ddl-pbp2b chromosomal region. PMID- 12089674 TI - Distribution of Legionella species and serogroups isolated by culture in patients with sporadic community-acquired legionellosis: an international collaborative survey. AB - This international collaborative survey identified culture-confirmed legionellosis in 508 patients with sporadic community-acquired legionellosis. Legionella pneumophila constituted 91.5% of the isolates. Serogroup 1 was the predominant serogroup (84.2%), and serogroups 2-13 (7.4%) accounted for the remaining serogroups. The Legionella species most commonly isolated were L. longbeachae (3.9%) and L. bozemanii (2.4%), followed by L. micdadei, L. dumoffii, L. feeleii, L. wadsworthii, and L. anisa (2.2% combined). L. longbeachae constituted 30.4% of the community-acquired Legionella isolates in Australia and New Zealand. PMID- 12089675 TI - Reduced lymphocyte-mediated antifungal capacity in high-risk infants. AB - Premature and critically ill infants are highly susceptible to Candida albicans. This study evaluated the lymphocyte-mediated antifungal capacity of infants relative to birth weight, prematurity, and illness severity. Growth inhibition of C. albicans by lymphocytes from preterm and low-birth weight infants was significantly reduced, compared with full-term and normal-weight infants. Lymphocyte growth inhibition of C. albicans is dependent on cell adhesion to the fungus. Compared with full-term infants, lymphocytes from preterm infants had a reduced capacity to adhere to C. albicans. Furthermore, infants with greater severity of illness (score for neonatal acute physiology [SNAP], >or=10) exhibited significantly reduced lymphocyte-mediated antifungal capacity and fungal adhesion. Although gestational age, birth weight, and SNAP were significantly associated with lymphocyte-mediated growth inhibition and adhesion, stepwise regression analysis demonstrated that gestational age best predicted both lymphocyte growth inhibition of and adhesion to the fungus. PMID- 12089676 TI - Prophylactic administration of liposomal amphotericin B is superior to treatment in a murine model of invasive aspergillosis after hematopoietic cell transplantation. AB - With use of a novel model of invasive Aspergillus fumigatus, the efficacy of prophylactic versus therapeutic administration of liposomal amphotericin B (L AmB) was tested in C57BL/6 mice. After lethal irradiation and transplantation of whole bone marrow (d 0), animals were challenged with conidia either intravenously or via nasal instillation on d +3 and divided into 3 groups: group I received 5% dextrose in water throughout the study period; group II received L AmB, 5 mg/kg, beginning on d +4; and group III received L-AmB, 5 mg/kg on d -4, d -2, d 0, and d +2, then daily starting d +4. Groups I and II did not survive intravenous challenge, whereas group III had a 40% survival rate. After nasal instillation of conidia, the survival was 25%, 35%, and 85% for mice in groups I, II, and III, respectively. These results demonstrate that prophylactic administration of L-AmB increased early survival against lethal challenge with A. fumigatus, compared with therapy instituted after infection. PMID- 12089677 TI - Radical cure of experimental cutaneous leishmaniasis by the bisphosphonate pamidronate. AB - The effects in vivo of the bisphosphonate drug pamidronate, used in bone resorption therapy, were investigated in an experimental model of cutaneous leishmaniasis. Pamidronate at an intraperitoneal dose of 10 mg/kg/day for 5 days effects a radical cure of cutaneous leishmaniasis in Balb/c mice, as evidenced by long-term disappearance of lesions; disappearance of amastigotes in lesion sites, as determined by histopathological analysis and cultivation of material obtained from lesions; and polymerase chain reaction analysis of necropsy material, using probes specific for kinetoplast DNA. Pamidronate is, therefore, a new lead compound for the synthesis of drugs effective against cutaneous leishmaniasis. PMID- 12089678 TI - Is the poorer rate of survival among patients with human immunodeficiency virus infection and anemia linked to immune activation? PMID- 12089680 TI - Reduced levels of CD4 cell spontaneous apoptosis in human immunodeficiency virus infected patients with discordant response to protease inhibitors. PMID- 12089681 TI - Tumors: patients and surgeons. PMID- 12089682 TI - Nerve growth factor is expressed in rat femoral vein. AB - PURPOSE: Entubulization is a well known method of nerve repair for defects too large to be reconstructed by direct suturing without tension. Vein grafts and alloplastic tubes have been used for entubulization in peripheral and cranial nerves, but the mechanism by which they promote healing is poorly understood. The overall hypothesis of this laboratory is that nerve growth factor (NGF) plays an important role in nerve regeneration after entubulization with a vein graft. The purpose of this pilot study was to localize NGF protein expression in the rat femoral vein. MATERIALS AND METHODS: Sciatic nerves and femoral veins were harvested from adult male Sprague-Dawley rats. Femoral arteries were also collected and used for comparison and validation of the analysis. All specimens were fixed in paraformaldehyde and embedded in paraffin. Specimens were either stained with hematoxylin and eosin or used for immunohistochemical reaction with anti-NGF antibody. RESULTS: Sciatic nerve was used as a positive control to identify the monofascicular architecture with hematoxylin and eosin and to document the positive immunohistochemical reaction. NGF immunoreactivity was present in the tunica intima and tunica adventitia of femoral vein and artery but not in the tunica media. CONCLUSION: The results of this pilot study indicate that NGF is detectable in both the intimal and adventitial layers of the rat femoral vein and artery but not in the smooth muscle wall. These findings suggest that vein grafts could potentially promote nerve regeneration by supplying NGF to the injured nerve. PMID- 12089684 TI - Changes in the reducing power of synovial fluid from temporomandibular joints with "anchored disc phenomenon". AB - PURPOSE: In this study, we examined the hypothesis that uncontrolled oxidative stress causes collapse of the lubrication system, which is considered a major initiator of temporomandibular joint (TMJ) dysfunction. The oxidative stress was evaluated by measuring the overall antioxidant capacity of the low-molecular weight antioxidants in the TMJ, using cyclic voltammetry (CV), in synovial fluid from normal and anchored disc phenomenon (ADP) TMJs. MATERIALS AND METHODS: Synovial fluids samples were taken from 13 normally functioning and 33 ADP TMJs. The samples were frozen initially on collection and analyzed in CV to measure their overall reducing power. RESULTS: CV measurements of the fluids collected from 90% of the healthy joints showed an anodic waves at peak potential [Ep(a)1/2] of 290 +/- 30 mV. Of the samples 56% showed another wave at 650 +/- 100 mV. These waves were generally absent in the fluid collected from ADP TMJs, but 2 new waves at 465 +/- 90 mV and greater than 750 mV were detected in 68% and 87% of the patients, respectively. CONCLUSIONS: The results show more anodic waves, most of which of higher potentials (greater than 750 mV) in ADP TMJs, indicating that the capacity to cope with oxidative stress is lower in these joints. It is not clear whether this is due to absence of low-molecular-weight antioxidants or their consumption by uncontrolled production of reactive oxygen species, which might be the initial step in the collapse of the lubrication system. PMID- 12089685 TI - Interleukin-1beta stimulates interleukin-8 production and gene expression in synovial cells from human temporomandibular joint. AB - PURPOSE: The aims of the present study were to isolate and characterize cultured synovial cells from human temporomandibular joint (TMJ) specimens and to investigate the effect of interleukin (IL)-1beta on IL-8 production and gene expression in those cells. MATERIALS AND METHODS: Synovial cells (HTS cells) were isolated from TMJ synovial tissues using an outgrowth method and then primary cultured. The cells were examined for cell-specific markers of fibroblast, macrophage, and dendritic cells using immunocytochemistry. HTS cells were then treated with IL-1beta, and amounts of IL-8 were measured by enzyme-linked immunosorbent assay. IL-8 production and expression were also investigated using immunocytochemistry and a reverse transcription-polymerase chain reaction method. RESULTS: HTS cells were positive for the fibroblast-specific markers, such as vimentin and propyl 4-hydroxylase. The macrophage or dendritic cell markers and HLA class II antigen were negative. Furthermore, IL-1beta enhanced IL-8 production in HTS cells in a time- and dose-dependent manner and stimulated IL-8 gene expression. CONCLUSIONS: HTS cells may provide important advantages for studies of the cellular and molecular mechanisms in the TMJ. In addition, we found that IL-1beta stimulated IL-8 production through an increase in IL-8 gene expression in HTS cells, which may be associated with the increase of infiltrating inflammatory cells seen in the synovial membrane of TMJ disorders. PMID- 12089686 TI - Recurrent neck disease in oral cancer. AB - PURPOSE: The goals were to examine the clinical and pathologic features of patients who developed metastatic squamous cell carcinoma in the cervical lymph nodes after initial treatment and to identify any common patterns. PATIENTS AND METHODS: A retrospective analysis of 35 patients of varying initial tumor stage was performed. There were 18 patients who had an initial neck dissection and 17 patients whose neck was managed by a "watch and wait" policy. RESULTS: Recurrence frequently involved level II nodes, and extracapsular spread was invariably present. The time taken for recurrence to develop was the same in both groups of patients (15 months, P =.35), and the overall median survival time after recurrence was 18 months (12 to 25 months, 95% confidence interval). In 27 of 29 patients (93%) who had the primary tumor resected, the thickness of tumor was greater than 5 mm. CONCLUSION: Neck recurrence may represent residual disease; it has histologically unfavorable features and consequently a poor prognosis. The frequency of recurrence at level II emphasizes the need for meticulous dissection in this region, and tumor thickness needs to be considered in planning treatment of the clinically negative neck. PMID- 12089688 TI - The surgical treatment of central giant cell granuloma of the mandible. AB - PURPOSE: The objective of this study was to report and evaluate our experience in the surgical treatment of mandibular central giant cell granuloma by resection without continuity defect and peripheral ostectomy. METHODS: A retrospective analysis was conducted of patients with central giant cell granuloma of the mandible who were treated between 1991 and 2000, in the Oral and Maxillofacial Surgery Unit at Jordan University of Science and Technology. A uniform surgical technique was used in all cases. The compact bone composed of the lower border of the mandible and/or posterior border of the ascending ramus, together with the nutrient periosteum attached to it, was preserved. All soft tissues in contact with or overlying the lesion and a margin of cancellous bone related to the lesion were excised. All patients were reviewed annually for a follow-up period of 1 to 9 years (mean, 3.9 years). RESULTS: Eighteen patients with central giant cell granuloma were included, (9 males and 9 females). Their age ranged from 10 to 46 years, with 89% younger than 40 years. Five (28%) lesions were in the incisor-canine region, 2 (11%) were confined to the premolar region, 4 (22%) were in the premolar-molar region, and 7 (39%) were in the molar-ramus region. All patients had aggressive central giant cell granulomas with pain, tooth mobility, and rapidly enlarging swelling. The initial diameter of lesions ranged from 2.7 to 10 cm. During the follow-up period, there was 1 case of recurrence, 2 (11%) patients had permanent lower lip paraesthesia, and no patient had obvious facial deformity. CONCLUSION: Our results suggest that resection without a continuity defect and peripheral ostectomy is a satisfactory method in the treatment of central giant cell granuloma of the mandible, with no or a very low recurrence rate and favorable postoperative function. PMID- 12089689 TI - Ameloblastoma in children. AB - PURPOSE: The purpose of this report was 1) to report the experience of the University of Maryland, Department of Oral and Maxillofacial Surgery (OMS Department) in the treatment of ameloblastoma in children and 2) to review the world literature on the treatment of ameloblastoma in children from 1970 to 2001. METHODS AND MATERIALS: This study first reviews the experience of the OMS Department of the University of Maryland with ameloblastomas in children and then reviews the literature on this subject. The first part of the study was undertaken by a retrospective chart review of all patients with a diagnosis of ameloblastoma in the OMS Department between May 1991 and December 1999. The literature on ameloblastoma in Western societies and Africa was separately reviewed from 1970 through 2001. Reports earlier than 1970 were not reviewed, as the histologic diagnosis of ameloblastoma was not well defined before that period. RESULTS: In the Maryland series, 11 patients under the age of 20 years with ameloblastoma were treated. Eight patients were seen primarily, and 3 presented with recurrent lesions. The average age was 15.5 years; 5 of 11 patients were black, and 9 of 11 tumors were unicystic ameloblastomas. The literature review showed 85 children in the Western reports and 77 reported from Africa. The average ages were 14.3 and 14.7 years, respectively, but unicystic ameloblastomas accounted for 76.5% of the Western and only 19.5% of the African children, with an increased frequency of occurrence in the mandibular symphisis in African (44.2%) versus Western (5.8%) patients. Analysis of recurrence after enucleation of unicystic ameloblastomas in 20 children followed at least 5 years or until recurrence showed a recurrence of 40%. CONCLUSIONS: Ameloblastomas in children differ from adults, with a higher percentage of unicystic tumors. African children appear to resemble the adult pattern. Although enucleation has been claimed to give acceptable recurrence rates in unicystic ameloblastoma, there are no large series with long follow-up in children. The histologic pattern that exhibits mural invasion in unicystic ameloblastoma suggests that more aggressive surgery is necessary. PMID- 12089692 TI - Analysis of human leukocyte antigens in patients with internal derangement of the temporomandibular joint. AB - PURPOSE: Spondyloarthropathy includes the subcategory of reactive arthritis (ReA). Spondyloarthropathies are commonly associated with certain human leukocyte antigen (HLA) alleles. Because we identified bacteria associated with ReA within the temporomandibular joint (TMJ), we now evaluate the frequency of HLA alleles in patients with TMJ pathology. PATIENTS AND METHODS: HLA typing of 129 patients (121 females and 8 males) performed by standard microcytotoxicity technique. Thirty patients had only class I (HLA-A and -B loci) evaluated. Ninety-nine patients had both class I and class II (HLA-DR loci) evaluated. Identification of alleles at the C locus was not performed. The antigenic frequency in the study group was compared to US white control subjects using a 2-tailed Fisher's exact test with a Bonferroni multiple comparison adjustment. RESULTS: The following class I HLA alleles, -A1 (32%), -A2 (50%), -A3 (33%), -B7 (23%), -B14 (14%), -B35 (20%), and -B44 (36%), including the B7 cross-reactive group (CREG) (49%) and class II alleles -DR1 (25%) and -DR4 (34%), were found to have an increased frequency in our patient group. CONCLUSIONS: Our study shows an increased frequency of several alleles that have been previously associated with arthropathy, and the alleles of the B7 CREG, in patients with TMJ pathology. Patients with these alleles may have an increased risk for the development of internal derangement of the TMJ as a consequence of the bacterial/infectious agents and host interactions with the subsequent cytokine/inflammatory response being influenced by their HLA phenotype. PMID- 12089691 TI - Pain, function, and psychologic outcome before, during, and after intraoral tumor resection. AB - PURPOSE: The aim of the present study was to determine the quality of life of patients with respect to pain before, during, and after ablative intraoral tumor surgery. PATIENTS AND METHODS: The Bochum Questionnaire on Rehabilitation was used to determine 147 items including the morphologic, functional, and psychosocial aspects of rehabilitation. One thousand six hundred fifty-two of 3,500 patients (47.2%) (413 women and 1,239 men) completed the questionnaire. The statistical investigation was carried out using 2-sample (independent) and paired (dependent) t tests. RESULTS: Pain and the functional impairment of chewing and swallowing are the most important parameters before treatment. Immediately after surgical treatment, other variables, such as speech intelligibility and mobility disorders in the head, neck, and shoulder regions, became more apparent. Pain was reported in the shoulder region in 38.5% and in the neck in 34.9% of patients after surgery. The temporomandibular joint was painful in 20.1%, the oral cavity in 18.7%, and the face in 8.2%. Pain was described by 7.4% of patients in other regions of the head. Seventy-five percent of 1527 patients were not taking pain medications during the investigation. The rate of physiotherapy consequently applied after surgery was less than 10%. CONCLUSIONS: Quality of life after ablative intraoral surgery is not greatly affected by pain. Because functional disorders play the dominant role in the impairment of postoperative quality of life, functional reconstruction, using microvascular techniques, and early rehabilitation, including physiotherapy, should be intensified. PMID- 12089693 TI - Assessment of patients treated for intracapsular fractures of the mandibular condyle by closed techniques. AB - PURPOSE: The goals to study different lines of intracapsular fractures of the mandibular condyle and to evaluate their influence on the prognosis after closed treatment. PATIENTS AND METHODS: Clinical, radiologic, and axiographic follow-up of 40 patients with 50 intracapsular fractures of the mandibular condyle was carried out after closed treatment. The examinations were performed an average of 22 weeks after treatment. Three types of intracapsular fractures were distinguished: type A, or fractures through the medial condylar pole; type B, or fractures through the lateral condylar pole with loss of vertical height of mandibular ramus, and type M, multiple fragments, comminuted fractures. RESULTS: Moderate to serious dysfunction was observed in 33% of the cases. Radiologic examination of fracture types B and M revealed a reduction in the height of the mandibular ramus of up to 13% compared with the contralateral side. These 2 fracture types also resulted in the most prominent deformations of the condylar head. Axiography revealed irregular excursions and a limitation of condylar movement in comminuted fractures of up to 74% compared with the nonfractured side. CONCLUSION: Lesions to the osseodiscoligamentous complex of the temporomandibular joint caused by intracapsular fractures of the mandibular condyle can be severe. The poor functional and radiologic results encountered in the fracture types B and M showed the limitations of closed functional treatment. PMID- 12089695 TI - The transgingival approach for placement of distraction implants. AB - PURPOSE: Since 1997, distraction implants have been clinically used for alveolar ridge distraction and, later, for prosthetic treatment. While 63 patients have been treated by the authors by alveolar ridge distraction with distraction implants with an open approach, the aim of this study was to demonstrate a minimally invasive technique of distractor placement via a transgingival approach. PATIENTS AND METHODS: Twelve patients were treated with a modified surgical incision using distraction implants. A tissue punch was used to remove transgingival mucosa, and a segmental osteotomy was performed using a vestibular incision. The distracted segment was pedicled at the lingual and crestal mucoperiosteum. Distraction was carried out for 0.5 mm per day, divided into 2 to 4 turns per day. A distraction of 5 to 7 mm was performed. At the end of distraction, the distraction insert was changed into a definitive stable implant insert. Prosthetic treatment was performed 4 months after the distraction period with fixed superstructures. The follow-up was performed with the aid of dental radiographs, evaluation of peri-implant probing depths, and Periotest values (Siemans, Bensheim, Germany). RESULTS: The outcome of this technique showed minimal scarring of the gingiva with good aesthetic results, the clinical and radiologic findings were satisfying, and the Periotest values were negative at every examination. The rate of complications was low. CONCLUSION: Minimal scarring and good aesthetic and functional outcome resulted in patients with alveolar ridge distraction performed with a transgingival approach. PMID- 12089696 TI - Temporomandibular disorders in 19-year-old Korean men. AB - PURPOSE: The purpose of this study was to determine the prevalence and the nature of the relationships between 3 temporomandibular joint disease (TMD) symptoms and symptoms of associated structures. MATERIALS AND METHODS: This study was designed to rule out the effect from the uneven composition of the samples on TMD symptoms. The samples were collected from subjects who were of the same age, gender, district, and race. Nineteen-year old men (n = 27,978) were selected and investigated by means of questionnaires and clinical examinations. The prevalence of each symptom was studied and tried to determine the effects of the TMD associated signs on the TMD signs. The indices allocated to reflect the TMD signs and symptoms and the others were dichotomized for bivariate analysis. The predictor variables were headache, neck pain, referred pain, stress, past trauma history in the TMJ, past TMJ dislocation, bruxism, and clenching. The outcome variables were mouth opening limitation, TMJ pain on rest, and TMJ pain during function. RESULTS: The incidence of masticatory muscle stiffness was 17.8%; TMJ sounds, 14.3%; headache, 7.2%; neck pain, 13.5%; bruxism, 8.4%; and clenching, 9.9%. Stress occurred in 12.8%, past trauma history in 11%, and previous joint dislocation in 2.5%. The experience of dislocation in the TMJ was found to be the most important risk factor in terms of mouth opening limitation (odds ratio, 4.08, P <.0001), joint pain during function (odds ratio, 5.50, P <.0001), and joint pain in the rest state (odds ratio, 4.63, P <.0001). Referred pain and the experience of trauma in TMJ were the secondary risk factors in terms of joint pain and referred pain and the stress in terms of mouth opening limitation. Considering referred pain can be induced by TMD, stress may be more related to mouth opening limitation (odds ratio, 2.18, P <.0001), and the experience of trauma in TMJ may be more related to pain in the rest state (odds ratio, 2.56, P <.0001) and during function (odds ratio, 2.47, P <.0001). CONCLUSIONS: The prevalence of TMD signs and symptoms as determined by this examination was in accord with the findings in women or mixed samples of other workers. Prior experience of a dislocated disc was found to be the most risky factor in TMD. Stress was related to limitations of mouth opening, and the experience of trauma in the TMJ was found to be related to pain in the joint region. Bruxism may not be a direct risk factor in TMD, and the clenching habit found to be more harmful than bruxism. PMID- 12089697 TI - Lesion of floor of the mouth. PMID- 12089698 TI - The gene: the polymerase chain reaction and its clinical application. AB - Chromosomal DNA transfers and stores information regarding the structure and function of the cell. Genetic information, encoded within sequences of nucleotides that compose DNA, is grouped into functional units called genes. Genetic diseases are caused by changes in the chromosomal DNA, leading to a change in the quantity or function of the protein gene product. In the past, genetic diagnosis was limited by the availability of sufficient quantity and quality of DNA and the absence of an efficient amplification procedure. The polymerase chain reaction (PCR), an inexpensive, rapid, and accurate means of amplifying DNA, is already making a major contribution to the diagnostic sciences. PCR techniques have been widely used in diverse applications, including molecular analysis of microbial pathogens, inheritable diseases and syndromes, and neoplasms. The purpose of this article is to 1) Review gene structure and function, 2) review principles of PCR technology and its applications in molecular biology, and 3) discuss an experimental clinical application of PCR to identify novel infectious agents responsible for odontogenic infections. PMID- 12089699 TI - Buccinator myomucosal island flap for postablative maxillofacial reconstructions: a report of 4 cases. PMID- 12089700 TI - Peripheral primitive neuroectodermal tumor of the maxillary gingivae with metastasis to cervical lymph nodes: report of a case. PMID- 12089701 TI - Bilateral mandibular cyst: lateral radicular cyst, paradental cyst, or mandibular infected buccal cyst? Report of a case. PMID- 12089702 TI - Postirradiation angiosarcoma of the head and neck: report of a case. PMID- 12089703 TI - Management of florid cemento-osseous dysplasia of the mandible producing solitary bone cyst: report of a case. PMID- 12089704 TI - A simple retrieval technique for accidentally displaced mandibular third molars. PMID- 12089705 TI - Fat transfer gun used as a precision injection device for injectable soft tissue fillers. PMID- 12089706 TI - Integration of the medical degree in oral and maxillofacial surgery: a 10-year follow-up. PMID- 12089707 TI - Recurrence rates in oral cancer. PMID- 12089708 TI - Recurrence of primary sclerosing cholangitis after liver transplantation. AB - Orthotopic liver transplantation (OLT) has become the only effective therapeutic option for patients with end-stage liver disease caused by primary sclerosing cholangitis (PSC). Excellent long-term outcome has been reported, with 5-year patient survival rates of approximately 80%. In the last few years, increasing evidence has emerged that PSC recurs after OLT. The diagnosis of PSC is based on well-defined cholangiographic features combined with biochemical and histological findings. However, none of these features is specific for PSC, particularly after OLT, because biliary strictures in the liver allograft can occur from a variety of causes other than recurrence. Therefore, PSC recurrence remains a controversial issue, especially because of a lack of a gold standard for diagnosis and well-established diagnostic criteria. Some reports provided cholangiographic evidence that post-OLT biliary strictures occurred more frequently in patients with PSC than in those who underwent OLT for other liver diseases (including patients with a Roux-en-Y biliary reconstruction). Because no other possible cause of biliary strictures could be invoked to explain the greater prevalence of these strictures, recurrent disease has been implicated. There also is histological evidence suggesting that PSC recurs after OLT. Histological findings suggestive of PSC were found more often in PSC allografts compared with a control group. Furthermore, histological features typical for PSC (fibro-obliterative lesions) were seen exclusively in liver biopsy specimens from patients with PSC. Recurrence of PSC was defined in a recent study from the Mayo Clinic by means of strict cholangiographic and histological criteria in a large cohort of patients with PSC in whom other causes of biliary strictures were excluded. PSC recurrence was found in 20% of patients. No risk factor for PSC recurrence could be found, and recurrent disease did not influence patient or graft survival after a mean follow-up of 4.5 years. In conclusion, several studies provided convincing evidence that PSC recurs after OLT, with an incidence of 5% to 20% and an interval to diagnosis of at least 1 year after OLT. To date, patient and graft survival do not appear to be negatively affected by disease recurrence in the intermediate term of follow-up. PMID- 12089709 TI - Cigarette smoking is associated with an increased incidence of vascular complications after liver transplantation. AB - Hepatic artery thrombosis (HAT) and other vascular complications are significant causes of morbidity after liver transplantation. Although cigarette smoking increases the risk of vascular complications after renal transplantation, its impact after liver transplantation remains unknown. Between May 1995 and April 2001, 288 liver transplantations were performed in 263 patients. Vascular complications developed in 39 patients (13.5%) (arterial complications, 28 patients [9.7%]; venous complications, 11 patients [3.8%]). Patient demographics, comorbid illnesses, and risk factors were analyzed using the Mann-Whitney U test, Chi-squared test, and Fisher's exact test. In patients with a history of cigarette smoking, incidence of vascular complications was higher than in those without history of cigarette smoking (17.8% v 8%, P =.02). Having quit cigarette smoking 2 years before liver transplantation reduced the incidence of vascular complications by 58.6% (24.4% v 11.8%, P =.04). The incidence of arterial complications was also higher in patients with a history of cigarette smoking compared with those without such history (13.5% v 4.8%, P =.015). Cigarette smoking cessation for 2 years also reduced the risk of arterial complications by 77.6% (21.8% v 5.9%, P =.005). However, the incidence of venous complications was not associated with cigarette smoking. Furthermore, there was no significant association between development of vascular complications and all other characteristics studied. Cigarette smoking is associated with a higher risk for developing vascular complications, especially arterial complications after liver transplantation. Cigarette smoking cessation at least 2 years before liver transplantation can significantly reduce the risk for vascular complications. Cigarette smoking cessation should be an essential requirement for liver transplantation candidates to decrease the morbidity arising from vascular complication after liver transplantation. PMID- 12089710 TI - Cigarette smoking--association with hepatic artery thrombosis. PMID- 12089711 TI - Predicting the decrease of conjugated bilirubin with extracorporeal albumin dialysis MARS using the predialysis molar ratio of conjugated bilirubin to albumin. AB - Extracorporeal albumin dialysis with the molecular adsorbent recirculating system (MARS) machine is a new supportive intervention for patients with liver failure. It removes bilirubin and other albumin-bound toxins from the patient and has been shown by preliminary studies of liver failure patients to be beneficial. Our study examines the ability of predialysis molar ratio of bilirubin to albumin to predict the decrease of bilirubin by MARS. We had 5 patients and results from 29 treatments. The results showed a significant correlation between the predialysis molar ratio of bilirubin (total and conjugated) to albumin to the reduction in bilirubin (total and conjugated): R(2) = 0.27 and 0.62 respectively, P <.005 for both. There was no significant correlation with the predialysis molar ratio of unconjugated bilirubin to albumin to the reduction in unconjugated bilirubin. The ratio of change in total bilirubin (micromol/L) to the predialysis molar ratio of total bilirubin to albumin and the ratio of change in conjugated bilirubin (micromol/L) to the predialysis molar ratio of conjugated bilirubin to albumin were 6.2 (+/- 4.2) and 10.8 (+/- 4.3), respectively (mean (+/- SD)). The results enable us to predict the likely reduction in bilirubin (especially conjugated) after each MARS treatment and also suggest to us that albumin infusion before MARS treatment may reduce the efficacy of bilirubin removal. Whether this ratio applies to other albumin-bound toxins remains open for investigation. PMID- 12089712 TI - Glomerular filtration rate assessment in individuals after orthotopic liver transplantation based on serum cystatin C levels. AB - Individuals after orthotopic liver transplantation (OLT) often show renal dysfunction, which may substantially affect the post-OLT course. Renal function after OLT is commonly assessed by means of serum creatinine (S(cr)) concentration or renal creatinine clearance (C(cr)). A glomerular filtration rate (GFR) estimate based on S(cr) level is not accurate enough because even a more marked decrease in GFR need not be associated with an increase in S(cr) level, especially in jaundiced patients. The study intends to try to estimate GFR in individuals after OLT by means of determining serum cystatin C (S(cyst)) concentrations. In 58 individuals (mean age, 49 +/- 7 years; 31 men, 27 women) at various intervals from OLT (mean, 14 +/- 10 months), GFR was estimated by using simultaneous determinations of S(cyst), S(cr), C(cr), and renal inulin clearance (C(in)). In most subjects (91.3%), C(in) was decreased to less than the lower limit of normal (80 mL/min/1.73 m(2)). A significant correlation (r = 0.70; P <.001) was found between 1/S(cyst) and C(in). Receiver operating characteristic analysis was performed on S(cyst) and S(cr) using a C(in) cutoff value of 80 mL/min/1.73 m(2). The area under the curve for S(cyst) was 0.912 +/- 0.044, and that for S(cr), 0.899 +/- 0.049. There was no statistically significant difference between these values. The sensitivity for a S(cyst) level of 1.20 mg/L (upper limit of normal value) to detect a decrease in GFR (measured as C(in)) below the lower limit of normal (80 mL/min/1.73 m(2)) was 96.1%. The sensitivity of S(cyst) level was significantly greater (P <.01) than the sensitivity of S(cr) level for men and at borderline significance for women (P =.05). Findings support the assumption that a S(cyst) level less than 1.2 mg/L indicates with a high degree of probability (P <.001) that GFR is not decreased to less than the normal limit. S(cyst) assessment in individuals after OLT could be proposed as a confirmatory test of a decrease in GFR in individuals with normal S(cr) levels. PMID- 12089713 TI - A tale of two markers. PMID- 12089714 TI - Cytokine gene polymorphisms and acute human liver graft rejection. AB - Interindividual differences exist in the capacity to produce cytokines. It has been reported that levels of in vitro cytokine production measured after stimulated cell culture are associated with polymorphisms in cytokine genes. Moreover, a correlation between heart, kidney, liver, and lung graft rejection or survival with cytokine gene polymorphisms has been described. In the present study, we analyzed the association of gene polymorphisms in T helper subtype 1 (T(H)1-), T(H)2-, and regulatory-type cytokines with human liver allograft rejection. Patients who received a primary liver graft from 1992 onward and were seen at the transplant outpatient clinic since then were included on this study (n = 89). Patients were HLA typed routinely. Biopsy-proven acute rejection occurred in 41 of 89 patients. After informed consent, blood was collected and DNA was obtained. Using amplification-refractory mutation system polymerase chain reaction, the following cytokine gene polymorphisms were determined: IL-2+166, IL 2-330, IL-15+13689, IL-15-80, TNF-A-308, TNFd3, IFN-G+874 (T(H)1-type cytokines), IL-4+33, IL-4-590, IL-6-174, IL-10-592, IL-10-819, IL-10-1082, IL-13+2043, IL-13 1055 (T(H)2 type cytokines), TGF-B1+869, and TGF-B1+915 (regulatory-type cytokines). Univariate analysis showed that polymorphisms of IL-10-1082, TGF B1+869, and HLA-DR6 were significantly related to liver graft rejection. Multiple logistic regression analysis was used to assess which variables remained significantly predictive of acute rejection. Multivariate analysis showed that TGF-B1+869 and HLA-DR6 were independently associated with the occurrence of acute rejection. These findings suggest a role for the regulatory-type cytokine transforming growth factor-beta1 in human liver graft rejection. PMID- 12089715 TI - Volumetric analysis of liver segments in 155 living donors. AB - Right-lobe graft has been used most frequently for living donor liver transplantation in adult patients; however, some donors cannot donate their right lobe (according to the Healey and Scroy's terminology) because the remaining residual liver would be too small. A recent study suggested the possibility of right posterior segment graft in these donors. The purpose of this study was to evaluate the feasibility of right lobe or right posterior segment graft with a volumetric analysis. Liver volumetry by computed tomography was performed in 155 consecutive donors, and the volume of each liver segment was calculated. To confirm the reliability of volumetric examination, the estimated graft volume and the actual weight were compared. The average volume ratios of the left lateral segment, left medial segment, caudate lobe, right anterior segment, and right posterior segment were 17%, 14%, 2%, 37%, and 30%, respectively. In 39 donors (25%), the volume ratio of the right lobe was over 70%. Of these donors, 72% had a larger (difference in volume ratio greater than 5%) right posterior segment than left lobe with caudate lobe. The relationship between the estimated volume and actual weight was linear. The present results suggest that right lobectomy carries a potential risk and that the right posterior segment may be useful as an alternative graft from the point of donor's safety. PMID- 12089716 TI - Long-term survival and late graft loss in pediatric liver transplant recipients- a 15-year single-center experience. AB - Increasing numbers of children undergo successful liver transplantation. Limited data exist on long-term survival and late graft loss. Survival and graft loss were studied in 376 primary liver graft recipients who survived more than 3 months after transplantation (80.5% of all primary graft recipients). Patient records were reviewed retrospectively for causes of graft loss. Risk factors were identified by analyzing graft, recipient, and posttransplant variables using multivariate Cox regression. One-, 5-, and 10-year actuarial graft survival rates in the study population were 94.6%, 87.3%, and 86.3%, respectively. Corresponding patient survival rates were 95.7%, 91.4%, and 90.4%. Forty-seven (12.5%) grafts were lost subsequently, 15 by patient death with preserved graft function. Survival rate after late retransplantation was 63.3%. Causes of late graft loss were infection (21.2%), posttransplant lymphoproliferative disease (PTLD, 21.2%), chronic rejection (17%), biliary complications (14.8%), and recurrence of malignant disease (8.5%). Independent risk factors for late graft loss and patient death included liver malignancy as primary disease, steroid resistant rejection, and PTLD. Graft loss rate was significantly increased for reduced-size grafts. Patients undergoing transplantation after 1991 and recipients of full size grafts were more likely to survive. In conclusion, the long-term outcome for pediatric primary liver graft recipients surviving the early postoperative period is excellent except for patients with liver malignancy. There is no increased risk of late graft loss with the use of split or living related donor grafts. Technical complications are only a minor factor in late graft loss, but complications related to immunosuppression and infection remain a major hazard and must be addressed. PMID- 12089717 TI - Treatment of posttransplantation recurrence of hepatitis C with interferon and ribavirin: lessons on tolerability and efficacy. AB - Recurrence of hepatitis C virus (HCV) infection after orthotopic liver transplantation is a major cause of graft failure. The aim of our study was to determine the safety, efficacy, and tolerability of combination therapy with interferon and ribavirin in the treatment of recurrent hepatitis after liver transplantation. Twenty-six patients (18 men) with histologically established HCV recurrence after liver transplantation for cirrhosis secondary to chronic HCV infection were treated with a combination of interferon alfa-2b (3 million units three times weekly) and ribavirin (800 to 1,000 mg/d). Dosage modifications were according to a standard protocol incorporating laboratory values and clinical side effects. Fifty percent of patients completed 1 year or more of therapy. On an intention-to-treat basis, nine patients (35%) showed an end-of-treatment virological response. Six of these nine patients completed greater than 6 additional months of follow-up, and all have had sustained virological responses. A histological response (decrease in histological activity index > or = 2) was seen in 75% of virological responders and 67% of nonresponders. Adverse events requiring dose modification or cessation of therapy occurred in 66% of patients. Adjuvant therapies used to support hemoglobin levels included erythropoietin and red blood cell transfusions. There were no independent pretreatment predictors of a virological response, perhaps because of the small sample size. Combination therapy with interferon and ribavirin may be beneficial in patients with recurrent HCV after liver transplantation. The majority of patients require dose modifications because of side effects. Histological response is common in virological nonresponders. PMID- 12089718 TI - Spectral electroencephalogram analysis in hepatic encephalopathy and liver transplantation. AB - The aim of this study is to evaluate the role of spectral electroencephalogram (EEG) analysis ((S)EEG) in quantitating brain dysfunction in cirrhotic patients, showing conditions of minimal hepatic encephalopathy (HE), and determining the impact of orthotopic liver transplantation (OLT) on its correction. (S)EEG was compared with visual EEG ((V)EEG) in 44 cirrhotic patients waiting for OLT and 44 healthy controls. Eighteen patients had overt HE, and 26 patients had no apparent HE. Twenty-one transplant recipients were reexamined 6 months after OLT. Computerized (S)EEG was performed by mean dominant frequency (MDF) and the occipital alpha-theta ratio, expressed as its logarithmic transformation (LogR). All patients underwent psychometric assessment. MDF and LogR correlated significantly with Child-Pugh score (P <.05) and the presence of HE (P <.0001). (S)EEG and (V)EEG determined minimal HE in 8 (31%) and 6 (23%) of 26 patients without overt HE, respectively. (S)EEG did not correlate with age, sex, cause of liver disease, portal hypertension, or psychometric test results. MDF and LogR improved in many transplant recipients. LogR was significantly lower in OLT candidates who died before OLT compared with OLT survivors. In conclusion, (S)EEG provides reliable quantitative information to evaluate the degree of HE and appears more sensitive than (V)EEG to discriminate a subclinical stage of HE. The improvement in (S)EEG results observed in transplant recipients confirms the reversibility of bioelectric brain dysfunction with restoration of liver functions. PMID- 12089719 TI - Bleeding complications from hepatic mucoidal aneurysmata: value of color duplex sonography after liver transplantation. PMID- 12089721 TI - Serial magnetic resonance imaging of central pontine myelinolysis. AB - Central Pontine Myelinolysis (CPM) is a rare neurologic complication after liver transplantation. The true incidence of CPM after orthotopic liver transplantation remains an estimate. However, with the introduction of magnetic resonance imaging, early recognition became feasible. In this report, we present a case of rapid resolution of CPM followed by serial magnetic resonance imaging scans. PMID- 12089720 TI - Recurrent portal hypertension after composite liver/small bowel transplantation. AB - Late technical complications of composite liver/small bowel transplantation procedures are often complex and have not been well defined. Here we describe the unusual presentation and management of two cases of recurrent thrombocytopenia due to hypersplenism resulting from portacaval shunt stenosis. Both patients presented with portal hypertension late after composite liver/small bowel transplantation. One patient presented with recurrent bouts of upper gastrointestinal hemorrhage and was ultimately found to have a stenosis of her native portacaval shunt. After unsuccessful balloon dilatation of the anastomosis, a successful side-to-side distal splenorenal shunt was performed. The second patient presented with severe thrombocytopenia, the etiology of which was determined to be a short segment occlusion of the inferior vena cava between the native portacaval shunt and the piggyback outflow anastomosis of the liver graft. Total caval occlusion prevented balloon dilatation; the patient was relisted for transplantation but died of chronic rejection four months later. Recurrent portal hypertension is challenging in patients who have had combined liver/small bowel transplantation. Surgeons performing intestinal transplantation need to be increasingly aware of these possible late complications. PMID- 12089722 TI - Hepatitis C quasispecies and severity of recurrence: cause, consequence, or coincidence? PMID- 12089723 TI - E-cadherin immunohistochemical analysis of histiocytoid carcinoma of the breast. AB - Histiocytoid carcinoma is a rare type of invasive breast carcinoma. It has been considered to be a variant of lobular carcinoma, a variant of apocrine ductal carcinoma, and an apocrine variant of lobular carcinoma and to resemble lipid rich carcinoma. In attempts to elucidate its histogenesis, investigators have used mucin and oil red O histochemical analysis and GCDFP-15 immunostaining. E cadherin is a relatively recent addition to the armamentarium of immunohistochemical markers used for cell differentiation and is a member of a family of transmembrane glycoproteins that has been shown to have a strong correlation with the histologic phenotypes of breast carcinoma. Most ductal carcinomas show diffuse membrane expression of E-cadherin, and lobular carcinomas are characterized by complete lack of membrane staining of E-cadherin. The object of this study was to use E-cadherin immunohistochemical analysis to help clarify the histogenesis of histiocytoid carcinoma. Fourteen cases containing the diagnosis of histiocytoid carcinoma of the breast were identified at M. D. Anderson Cancer Center (Houston, TX) from 1988 to 2001. All cases were rereviewed, histologic features were evaluated, and immunohistochemical staining with E-cadherin and GCDFP-15 was performed. Clinical information was extracted from the patients' medical records. Eleven cases met published histologic criteria for histiocytoid carcinoma. The remaining three cases were apocrine carcinoma. The pattern of tumor infiltration was solid, without secondary lumen formation in all cases of histiocytoid carcinoma. Lobular carcinoma in situ was identified in eight cases, but was absent in three. There was no E-cadherin immunohistochemical staining in eight of the 11 cases of histiocytoid carcinoma (72.7%). GCDFP-15 was immunoreactive in all 10 cases of histiocytoid carcinoma where it was performed. Follow-up data was available for nine of the 11 cases of histiocytoid carcinoma: six patients were alive with disease at 1.5 to 48 months, one patient had died of disease at 60 months, and two patients had no evidence of disease at 32 and 45 months. We conclude that histiocytoid carcinoma has an immunophenotypical profile consistent with both ductal and lobular differentiation. Moreover, the lack of consistent morphologic features, a specific clinical profile, and a distinct immunohistochemical pattern lead us to hypothesize that histiocytoid carcinoma is not a special type of breast cancer. PMID- 12089724 TI - Cyclooxygenase-2 (COX-2) expression by immunohistochemistry in glioblastoma multiforme. AB - Cyclooxygenase-2 (COX-2) (an enzyme responsible for the conversion of arachidonic acid to prostaglandin) upregulation has been described in association with a variety of tumors including gliomas. This study reviews the immunohistochemical profile of 47 glioblastoma multiforme using COX-2 antibody and correlates the results with MIB-1 (marker of cell proliferation) immunostaining. Positive staining with COX-2 was observed in 35 tumors (74.5%); more than 10% tumor cell positivity was seen in 10 tumors (21.3%). Immunostaining results were as follows: no staining, N = 12 (25.5%); 0% to 5% of tumor cells, N = 20 (42.6%); 5% to 10%, N = 5 (10.6%); 10% to 20%, N = 4 (8.5%); 20% to 50%, N = 4 (8.5%); and greater than 50%, N = 2 (4.3%). The mean MIB-1 labeling index for all tumors ranged from 3.0 to 76.4 (mean, 19.6). Mean MIB-1 labeling indices were higher in tumors with greater than 5% COX-2 immunostaining (mean MIB-1 labeling index, 23.5) versus tumors with 0% to 5% COX-2 immunostaining (mean MIB-1 labeling index, 17.7). There is evidence of COX-2 expression by immunohistochemistry in the majority of glioblastoma multiforme. As a group, tumors with a higher rate of cell proliferation tended to have increased expression of COX-2. These findings are significant in that therapeutic agents, which inhibit COX-2, are currently available and may play a role in the management of glioblastoma multiforme. PMID- 12089725 TI - The utility of epithelial membrane antigen and vimentin in the diagnosis of chromophobe renal cell carcinoma. AB - We evaluated the immunohistochemical expression of epithelial membrane antigen (EMA) and vimentin (VMT) in chromophobe renal cell carcinoma (CHRCC). We also studied the utility of EMA and VMT immunostains in helping differentiate CHRCC from renal oncocytoma and conventional (clear cell) renal cell carcinoma with granular morphology (GCRCC). Immunohistochemical staining for EMA and VMT was performed on 21 cases of CHRCC, 16 cases of renal oncocytoma, and 28 cases of GCRCC. The diagnosis in all cases was by concurrence of all pathologists involved in the study and was based entirely on examination of routinely stained slides. All cases were classic examples of these tumor types and presented no diagnostic difficulties. The intensity of immunohistochemical staining was graded on a scale of 0 to 3 (0 = no staining; 1 = equivocal; 2 = unequivocal, moderate intensity; and 3 = unequivocal, high intensity). Positive immunohistochemical staining was defined as unequivocal staining of at least 20% of the neoplastic cells. All cases of CHRCC were positive for EMA and negative for VMT. The same immunophenotype was observed in 75% of renal oncocytoma and 21% of GCRCC. In summary, all CHRCC cases in our study demonstrated immunohistochemical staining for EMA and not VMT. However, we also found that the same immunophenotype is observed in 75% of renal oncocytoma and in 21% of GCRCC, precluding its utility for positive identification of CHRCC. Nevertheless, the lack of such an immunophenotype is a reliable indication that a neoplasm under consideration is not CHRCC. PMID- 12089726 TI - Dedifferentiated chondrosarcoma with a noncartilaginous component mimicking a conventional giant cell tumor of bone. AB - We report a case of dedifferentiated chondrosarcoma in which the dedifferentiated component of the tumor shows a close histologic resemblance to a conventional giant cell tumor of bone. The tumor affected a 30-year-old woman with a long history of left shoulder discomfort and limitation of motion. Radiographic studies revealed a biphasic destructive lesion in the left proximal humerus composed of high-signal lobulated component on T2-weighted magnetic resonance image accompanied by a low signal intensity component exhibiting destructive growth with extension into soft tissue. Microscopically, two different areas consisting of the chondroid tissue and nonchondroid giant cell-rich lesion resembling conventional giant cell tumor of bone were found. Considering that the prognosis and survival associated with these two entities are very different, it is important to be aware of this variant of dedifferentiated chondrosarcoma to avoid the misdiagnosis of conventional giant cell tumor of bone. PMID- 12089727 TI - Extensive myxoid change in well-differentiated papillary mesothelioma of the pelvic peritoneum. AB - We present a unique case of papillary mesothelioma of the pelvic peritoneum with extensive myxoid change in a 44-year-old woman. The patient presented with lower abdominal pain; imaging studies revealed a pelvic mass. Microscopic examination of the surgically resected specimen showed extensive areas of myxoid stroma and only focal areas of classical papillary mesothelioma. A small biopsy of this lesion might have been misinterpreted as a soft tissue neoplasm with myxoid stroma or pseudomyxoma peritonei. It is suggested that mesothelioma with myxoid change should be included with differential diagnoses of myxoid lesions of the peritoneum. PMID- 12089728 TI - Solitary fibrous tumor of the tongue: report of a case with immunohistochemical and ultrastructural studies. AB - Solitary fibrous tumors are rare in extrapleural sites and extremely rare in the oral cavity. We report a case of a solitary fibrous tumor arising from the tongue of a 70-year-old woman. The tumor measured 1.6 cm in maximum diameter and consists of spindle-shaped cells distributed in a haphazard pattern. Immunohistochemical studies show strong positivity for CD34 and bcl-2, and weak positivity for desmin. Smooth muscle actin and S-100 protein are negative. Electron microscopy shows uniform neoplastic spindle cells with mesenchymal features. The differential diagnosis for spindle cell neoplasms in the tongue is discussed. PMID- 12089729 TI - Composite mycosis fungoides and B-cell chronic lymphocytic leukemia. AB - Concordant or composite mycosis fungoides and B-cell chronic lymphocytic leukemia (B-CLL) is exceedingly rare, with only 10 cases previously described to our knowledge. We report a case of a 64-year-old woman who developed generalized erythroderma 5 years after the diagnosis of early stage B-CLL. Over the next 6 years of her clinical course multiple sequential samples of skin, peripheral blood, and one enlarged lymph node were studied in detail by flow cytometry, immunohistochemistry, molecular diagnostics, and electron microscopy. The progressive cutaneous infiltrates were initially characterized as leukemia cutis, infiltration by B-CLL. Three years later, when she developed worsening skin disease and lymphadenopathy, the cutaneous infiltrates were characterized as cutaneous T-cell lymphoma. At that point, a biopsy of an enlarged lymph node revealed a composite lymphoma of both B-CLL and cutaneous T-cell lymphoma, and the peripheral blood also contained circulating cells of both neoplasms. Herein we summarize the literature on concordant cutaneous T-cell lymphoma and B-CLL, and the literature on concordant T- and B-cell neoplasms in general, with a review of the postulated relationships between these neoplasms. PMID- 12089730 TI - Sclerosing extramedullary hematopoietic tumor. AB - A 73-year-old woman presented with abdominal pain, weakness, and weight loss. Pertinent medical history included myelofibrosis, severe anemia, and lumpectomy for breast cancer. Computed tomography showed marked splenomegaly and numerous soft tissue masses in the mesentery. Excisional biopsy of the mass showed an encapsulated tumor with yellow-tan nodular growth pattern. Microscopically, the tumor was characterized by the presence of bizarre giant cells in background of prominent dense fibrous stroma. The atypical giant cells were positive for FVIII antibody staining. Many other cells were positive for CD43 antibody staining, indicating that the tumor was hematopoietic in nature, so-called sclerosing extramedullary hematopoietic tumor. Extramedullary hematopoiesis is a common phenomenon associated with severe anemia of any cause. When associated with myelofibrosis, however, it has different morphology from extramedullary hematopoiesis of the other causes. The significance of recognizing sclerosing extramedullary hematopoietic tumor is that it is often mistaken for sarcomas and other malignant neoplasms, especially when the clinical history is unknown. PMID- 12089731 TI - The pancreaticoduodenectomy. PMID- 12089732 TI - Blastomycosis: The great pretender can also be an opportunist. Initial clinical diagnosis and underlying diseases in 123 patients. AB - Clinically, blastomycosis can be difficult to recognize even in the endemic areas where clinicians are aware of this problem. In only 18% of 123 patients from the University of Mississippi Medical Center (Jackson, MS) blastomycosis was correctly suspected at the initial patient evaluation. Pneumonia sensu latu (40%), malignant tumors (16%), and tuberculosis (14%) were the most common misdiagnoses. The false first impression frequently resulted in unnecessary surgeries or treatment delays, with patients receiving inefficient antibiotic therapy for months. The presence of cutaneous involvement by the disease makes its' recognition easier for the clinician, raising the percentage of correct initial diagnosis to 64%. To evaluate the association with immunodepression, the presence of other diseases was also searched among the 123 patients. An immunodepressive condition preceded the fungal disease in 25% of patients. Another associated disease commonly found in blastomycotic patients was diabetes mellitus (22%). Blastomycosis is correctly suspected at the first clinical evaluation in only a small percentage of patients; pneumonia, cancer, and tuberculosis are the most common clinical considerations. Cutaneous involvement leads the clinician to the correct diagnosis in the majority of cases. One fourth of the patients with blastomycosis had underlying immunodepressive conditions, and underlying diabetes mellitus is present in 22% of patients. PMID- 12089733 TI - Comparison of clinical outcomes and natural morphologic changes between sequestered and large central extruded disc herniations. AB - A prospective and longitudinal investigation concerning clinical outcomes and morphologic changes of large lumbar disc herniations by MR imaging. To compare the clinical outcomes and the natural morphologic changes of between sequestered and large central extruded disc herniations. The spontaneous disappearance or diminution of large herniated lumbar discs in the spinal canal is known. Poor clinical outcome and small changes of herniated discs have been shown for large central extruded disc herniations with conservative treatment. The study population consisted of 22 patients with sequestration and a large central as extrusion established by an MR imaging study. Seventeen (11 patients with sequestration, and 6 patients with a large central extrusion) patients underwent a follow-up MR imaging study. The size of the herniated disc was measured on serial MR imaging studies, and the changes in size were classified into four categories. Clinical evaluations were also performed using a visual analogue scale (VAS), the Oswestry low back pain disability questionnaire, the straight leg raising test (SLRT) and so forth. Both the sequestered and large central extruded disc herniations showed a successful clinical outcome after conservative treatment in 17 of 22 patients (77%) in total: 11 of 13 patients (85%) with sequestered disc herniations, and 6 of 9 patients (67%) with large central extruded disc herniations. VAS and Oswestry disability scoring showed a greater change in the group with sequestration than in the group with large central extrusions. In the group with sequestration, seven patients reported the disappearance of herniated disc materials, and four patients showed a marked decrease in the size of their herniated discs in follow-up MR images. However, in the group with large central extrusions, only two patients showed a decrease in the size of their herniated discs. Large central extruded disc herniations can be treated successfully by conservative treatment. Outcomes seemed to be as good as or slightly inferior to those of sequestered disc herniations. However, a greater morphologic decrease in the herniated discs occurred more frequently for sequestered disc herniations than for large central extruded disc herniations. PMID- 12089734 TI - Electrophysiological evaluation of the incidence of martin-gruber anastomosis in healthy subjects. AB - The Martin-Gruber Anastomosis (MGA) is probably the most well known of the anastomotic anomalies that occur at various levels between the median and ulnar nerves. It is formed by motor axons from the median nerve or its branch anterior interosseous nerve that cross in the upper forearm to join the ulnar nerve. The purpose of this study was to establish the frequency of MGA in healthy subjects and to draw the attention of clinicians working in the neurophysiological laboratory to the presence of this anastomosis, and thus to avoid possible misinterpretations of data from needle electromyography (EMG) and nerve conduction studies. 100 volunteers (60 women and 40 men) were selected for the study. Surface recording electrodes were placed on the right hand thenar, hypothenar and on the first dorsal interosseous (FDI) muscles. The median and ulnar nerves were stimulated supramaximally at the wrist and at the elbow and compound muscle action potentials (CMAPs) were recorded and their amplitudes evaluated. MGA was found in 27 of the 100 subjects. The type of anastomosis most frequently seen was type II, which was observed in 21 subjects. Type I anastomosis was observed in three, type I + type II in two and type III anastomosis in one subject. It can thus be concluded that MGA is frequently encountered and it should be borne in mind that abnormal innervation models may influence the electrophysiological findings and thus give rise to faulty interpretations, especially in the case of median and ulnar nerve lesions. PMID- 12089735 TI - Smooth muscle cells transplantation is better than heart cells transplantation for improvement of heart function in dilated cardiomyopathy. AB - Muscle cell transplantation may delay or prevent cardiac dilation in dilated cardiomyopathy. The present study was designed to compare the effects of the heart function of smooth muscle cell (SMCs) auto-transplantation and heart cell (CMs) allo-transplantation in dilated cardiomyopathic hamsters, and to determine which cells are better for cell transplantation. CMs and SMCs were isolated from BIO 53.58 hamsters, and cultured for transplantation. CMs, SMCs (4 X 10(6) cells each) or culture medium were transplanted into 17 weeks old BIO 53.58 hamsters to achieve CM transplantation (CMTx), SMC transplantation (SMCTx), and controls (Con) (N=10 each). Cyclosporine (5 mg/Kg) was administered subcutaneously to CMTx. Healthy hamsters (sham, N=6) were used to compare heart functions. Four weeks after transplantation, heart function was evaluated in all groups using a Langendorff perfusion apparatus. Histology demonstrated severe focal myocardial necrosis in the dilated cardiomyopathic hearts. CMTx and SMCTx formed huge muscle tissue in the dilated myocardium. Sham, SMCTx, and CMTx had a better heart function than Con (p < 0.01), and SMCTx had a better peak systolic pressure (p < 0.05) and developed pressure (p < 0.05) than CMTx at any balloon volume. However, sham and SMCTx were not statistically different. SMCTx and CMTx formed muscle tissue and produced better heart function in the cardiomyopathic hearts, and SMCTx showed better systolic and developed pressures than CMTx, even though they were similar in other functions. Significantly, SMCTx had heart functions, which were similar to those of healthy hamster's hearts. PMID- 12089736 TI - Expression of CD40 and Fas ligand in Bowen's disease, squamous cell carcinoma and basal cell carcinoma. AB - Impaired regulation of apoptosis is known to be associated with the development of various cancers, and Fas/Fas-ligand (FasL) is known to play an important role in apoptosis. CD40 is a cell surface receptor, which when ligated modulates apoptosis in some cell types. The expressions of CD40 and FasL were examined in 10 normal skins, 7 Bowen's disease skins, 10 squamous cell carcinomas (SCCs) and 12 basal cell carcinomas (BCCs) immunohistochemically. In the normal epidermis, CD40 was more highly expressed in the keratinocytes of the squamous cell and granular layers than in those of the basal layer, and FasL expression was observed in the cell membrane of keratinocytes at the basal and squamous cell layers. CD40 expression was significantly higher in SCCs than in normal or Bowen's disease skin, while FasL expression was significantly higher in Bowen's disease than in SCCs. BCCs expressed the lowest levels of CD40 and FasL. These results suggest that altered CD40 and FasL expression may be related with the progression of SCC, and the marked reduced expression of CD40 and FasL may explain the biologic behavior of BCCs. PMID- 12089737 TI - Surgical treatment of tuberculous cold abscess of the chest wall. AB - The incidence of tuberculosis (Tbc) infection is high in some parts of the world and tuberculous cold abscess of the chest wall (CACW) often fails to respond to medical treatment. Medical records of 178 patients who underwent surgical treatment of chest wall abscesses from July 1970 to Sept. 2000 were reviewed and 89 patients who were pathologically confirmed as Tbc cold abscess cases were included in this study. Their ages ranged from 9 to 71 years (mean 33.3 years) and the male to female ratio was 1.2:1 (49 male, 40 female). The symptoms were palpable chest wall mass, pain and pus discharge, and three patients had multiple lesions. Twenty-five patients (28%) underwent excision of chest wall abscesses and 64 patients (72%) underwent chest wall and rib resection. Tbc medication was given preoperatively in 39 patients for an average of 6.3 months and all patients were given Tbc medication postoperatively for an average of 12 months. Postoperative complications were bleeding, pus discharge, empyema, pleural effusion, wound dehiscence, subcutaneous emphysema and activation of pulmonary Tbc. The disease recurred in 7 patients (7.8%) and these 7 patients all underwent a second operation. We recommend preoperative Tbc medication and complete resection of chest wall abscesses including any suspicious ribs. Postoperative Tbc medication for a minimum of 12 months is essential to decrease the risk of a relapse. PMID- 12089738 TI - Surgical outcome of female genital fistula in Korea. AB - This purpose of this study was to establish a new standard for the surgical management of female genital fistula in Korea. From January 1992 to October 2001, 117 patients with female genital fistula who were admitted to the departments of obstetrics and gynecology, urology and general surgery were analyzed. Nine patients with congenital etiologies and 48 patients who were treated conservatively were excluded. The relationships between surgical outcome and the cause of fistula, the location of fistula, and the various surgical methods were analyzed. In spite of appropriate surgical treatment, fistulas due to cervix cancer management had the worst prognosis. In terms of location, fistula recurrence after surgical repair was most common in the bladder fundus and base. The transvaginal and transrectal approaches are suitable for fistulas located in the lower vagina. The transabdominal approach is appropriate for fistulas located in the functional portions such as the bladder and ureter, for fistulas which are difficult to expose surgically by either the vaginal or rectal approach, or in cases with severe adhesions. In cases of cervix cancer, extra care should be taken during surgical expiration or definitive radiotherapy, especially when the areas involved are the bladder fundus and base. The nature of the surgical approach should be decided by the location of the fistula, the functional importance of the area, and the degree of surgical exposure during the corrective procedures. PMID- 12089739 TI - The circadian rhythms of blood pressure and heart rate in the hypertensive subjects: dippers and non-dippers. AB - The purpose of this study was to compare the circadian blood pressure and heart rate rhythm of dippers and non- dippers. Biochemical and clinical characteristics of dippers and non-dippers were also compared to determine whether non-dippers have an increased prevalence of hypertension- related conditions. The subjects were 123 out-patients with essential hypertension who had undergone ambulatory blood pressure monitoring at the University Hospital in Inchon, South Korea, from January 1, 1997 to December 31, 1998. Daytime values were determined between 6:00 AM and 8:00 PM and the nighttime values were determined between 8:00 PM and 6:00 AM. Non-dippers were defined as those who showed a reduction in systolic blood pressure, diastolic blood pressure or heart rate less than 10% and they accounted for 25%, 32% and 31% of the subjects, respectively. The timings of the circadian systolic and diastolic blood pressure and heart rate in dippers were very consistent: showing the lowest values during the night, rising in the early morning and reaching a plateau in the late morning. Non-dippers' circadian rhythms of systolic and diastolic blood pressures fluctuated less than those of dippers, and the 24-hours heart rate rhythms of the dipper and non- dipper groups were completely reversed. None of the biochemical characteristics tested in the present study showed a significant difference between dippers and non-dippers whether dipper was classified by systolic or diastolic blood pressure or heart rate. Similarly, hypertension-related symptoms and complications were not associated with the dipper/non-dipper description, whether classified by sBP, dBP, or HR. PMID- 12089740 TI - Diagnostic criteria of internal jugular phlebectasia in Korean children. AB - Internal jugular phlebectasia (IJP) is a fusiform dilatation of the internal jugular vein (IJV), usually presented as a neck mass in children. Accurate diagnosis from carefully directed history, physical examination, and radiological study could result in lifesaving therapy. We performed our study to suggest possible clinical diagnostic criteria for IJP in Korean children. We reviewed three cases of IJP (patients group) and compared the diameter of the internal jugular phlebectasias with diameters of IJVs in ten normal children (control group) using ultrasonography (USG). There were no significant differences in the range of diameters in the resting state between the two groups. The diameters on the right side, compared with those on the left side, showed no statistical significance (p > 0.05). The range of expanding diameter and average expanding ratios (resting state to Valsalva maneuver x 100%) showed a statistical difference between the two groups (p < 0.05). PMID- 12089741 TI - Evaluation of HER2/neu status by real-time quantitative PCR in breast cancer. AB - Over-expression of the human epidermal growth factor receptor-2 (HER2/neu) has been observed in many cancers, and is associated with a poor prognosis. Recent adjuvant treatment with anti-HER2 monoclonal antibodies in breast cancer has increased the demand for an evaluation of the HER2/neu status in breast cancer. The aim of this study was to investigate the HER2/neu status in breast cancer by a real-time quantitative polymerase chain reaction (PCR) method using LightCycler (Roche Diagnostics, Mannheim, Germany). DNA samples from the fresh tumor tissues of 27 patients with breast cancer were analyzed in parallel using immunohistochemistry (IHC) and the other prognostic parameters including estrogen receptor, progesterone receptor, cytokeratin, and DNA ploidy. Ten (37%) out of 27 cases tested were positive for HER2/neu, while 16 (73%) out of 22 tested positive through an IHC study. The correlation between the DNA aneuploidy and the positive results for HER2/neu were only observed using the real-time PCR method (p < 0.05). There was no significant correlation between the HER2/neu status and the S phase fractions of the DNA ploidy or other parameters. This study demonstrated that there is marked discordance in the results for the HER2/neu status according to the various methods used. Real-time quantitative PCR for HER2/neu appears to be clinically useful due to its simplicity and ability to produce rapid results. PMID- 12089742 TI - Effect of intracavitary injection of urokinase in the experimentally induced early pyogenic liver abscess of the rabbit. AB - This study was carried out to assess the effects of intracavitary injection of urokinase in the early liver abscess (ELA) of the rabbits. ELAs were induced on 25 in 47 New Zealand rabbits, which were divided into two groups, with 15 in group A, and 10 in group B. Urokinase was injected into the ELA of group A, and normal physiologic saline into those of group B. One and a half hours after the injections, the rabbits were sacrificed and evaluated by pathologists for the degree of fibrosis of the ELA wall, and fibrinolysis in the ELA itself. Statistical analyses were performed between the two groups. The following ELA sizes for each group were obtained: Group A, 4.3 x 2.9-10.1 x 7.2 mm (mean 7.1 x 4.1 mm); Group B, 4.6 x 2.7-15.0 x 9.7 mm (mean 8.5 x 4.57 mm). Eleven (73%) in group A showed grade II fibrosis of ELA wall, and 8 (80%) in group B showed grade III fibrosis of ELA wall (p=0.002). On pathological analysis, 5 (46%) in group A showed grade II fibrin, and 8 (80%) in group B showed grade III fibrin, of the ELA (p=0.09). In conclusion, injection of urokinase, into the ELAs, can reduce the degree of fibrosis of abscess walls. PMID- 12089743 TI - Uterine arterial embolization for the treatment of uterine leiomyomas. AB - OBJECTIVE: The purpose of this study was to establish guidelines for using uterine artery embolization for the treatment of uterine leiomyomas accompanied by adenomyosis in Korea. This study was performed to investigate 1) the effects of uterine embolization on leiomyoma accompanied by adenomyosis, 2) the outcome of uterine embolization according to the embolization materials used, 3) the relationship between the Resistance Index (RI) and the change in uterine volume. MATERIALS AND METHODS: We performed a retrospective study on 37 women who had uterine leiomyomas accompanied by adenomyosis. Bilateral uterine artery embolization was performed over a period of 17 months in 37 patients (age range 25 - 65) experiencing pain, hypermenorrhea, urinary frequency, etc. due to leiomyomas. Ultrasound imaging was performed before the procedure and at mean 6.9 months after the procedure. RESULTS: All procedures were technically successful. Mean clinical follow-up time was 12.8 months. Minor complications occurred in 82% patients following the procedure. After follow-up imaging, the median uterine volume decreased by 34.4% and the dominant myoma volume decreased by 86%. There was no statistical difference in the uterine volume reduction and the dominant myoma size reduction, whether the occluding agent was polyvinyl alcohol, polyvinyl alcohol plus gelfoam, or gelfoam, and whether the ultrasound measured Resistance Index value before the procedure was low or high. CONCLUSION: Primary candidates for uterine artery embolization include those with symptomatic uterine leiomyomas, who no longer desire fertility, but who wish to avoid surgery or are poor surgical risks. According to the results of our study, adenomyosis should not be considered as a contraindication for uterine artery embolization. However, because little data is available about the subsequent reproductive potential following this procedure, it should not be routinely advocated for infertile women. Further investigation is warranted regarding the choice of occluding agent and the role of the Resistance Index. PMID- 12089744 TI - Diverse vacA allelic types of Helicobacter pylori in Korea and clinical correlation. AB - Helicobacter pylori has a diversity of vacA allelic types. The purpose of this study was to correlate the vacA status and the clinical outcome. After constructing specific primers for the vacA signal sequence, H. pylori-positive antral biopsy specimens were examined for the vacA status in 25 gastric ulcers, 31 duodenal ulcers, 22 gastric cancers, 42 chronic gastritis, and 8 gastroduodenal ulcers. The relationship between the vacA allele and the clinical disease was examined. The vacA genotype s1c/m1 is predominant in Korea (71/128, 55.5%). Other strains including s1b or s2 were not found in this study. s1c/m1 was more prominent in duodenal ulcers, than in gastric ulcers (p=0.041) and cancer (p=0.029). Seven out of 8 patients with gastric and coexistent duodenal ulcers had the s1c/m1 allele. No statistical differences in the positive rates of the s1a/m1, s1a/m2, and s1c/m2 alleles among the disease groups were found. In conclusion, s1c/m1 is the main vacA allele in Korea and it is particularly associated with duodenal ulcers. PMID- 12089745 TI - The effect of initial serum neuron-specific enolase level on clinical outcome in acute carotid artery territory infarction. AB - The prediction of functional outcome in patients with acute cerebral infarction depends on many factors. Various techniques have been applied to predict severity and outcome after cerebral infarction. Neuron-specific enolase (NSE) is a component of a specific brain enzyme and a useful marker of brain injury. We evaluated the relation between initial serum NSE level and short- and long-term clinical outcome in 59 patients with acute cerebral infarction and in 38 age matched healthy controls. Serum NSE levels were determined in patients with carotid artery (CA) territory cerebral infarction within 24 hours of onset. Brain MRI was performed four to seven days after stroke. Patients were divided into two groups: large CA territory infarction with a lesion extending cortex (cortex group), and small subcortical CA territory infarction (subcortical group) with a lesion confined to the subcortical white matter. We compared the initial serum NSE levels of the two groups. National Institute of Health Stroke Scale (NIHSS) was determined at admission and seven days after onset and the modified Rankin's scale was used at the 3 months follow-up after onset. Serum NSE levels were significantly elevated in patients with acute cerebral infarction compared with the normal controls (13.88 +/- 5.47 ng/dl vs. 8.15 +/- 1.53 ng/dl, p < 0.05). The initial (< 24 h) serum NSE level was higher in the cortical group than in the subcortical group (16.68 +/- 5.70 ng/dl vs. 10.98 +/- 3.34 ng/dl, p < 0.05). NIHSS on admission and on the 7th day correlated with the initial serum NSE level (p < 0.05), as were more severe functional outcomes, as determined 3 months after onset (p < 0.05). This study shows that initial serum NSE level may be a useful marker for severity in acute ischemic stroke, and that it may be well correlated with short-term and long-term functional outcomes. PMID- 12089746 TI - Normal values of total body water in healthy Korean adults: comparison with data from western populations. AB - To find reference total body water (TBW) values in healthy Korean adults, we performed single frequency bioelectrical impedance analysis on 2942 healthy adults and compared these data with those of normal western adults. Males were found to have greater TBW than females. Conversely, females were found to have greater percent fat (pFat) than males. In both sexes, body weight (BW) and TBW were significantly lower in age groups of < 30, 50 - 59, > or = 60 years than in the 40 - 49 years age group. pFats were significantly higher in age groups of 50 59 and > or = 60 years than in the 40 - 49 years age group. pFats were similar among age groups of < 30, 30 - 39, and 40 - 49 in both sexes. In all age groups, pFats were significantly higher in females than in males. TBW was significantly correlated with BW, height (HT), fat mass, and body mass index (BMI) in both sexes. There was a significant correlation between TBW and age in males(r=-0.15, p=0.00), but not in females (r=-0.02, p=0.45). On linear regression analysis, TBW values were independently associated with BW, HT and age in males and were independently associated with BW and BMI in females. In all age groups, Korean males and females had lower mean TBW than in Western populations. However, this difference was greater in males than in females. In summary, we represented the contemporary normal TBW data in healthy Korean adults. At present, there are no national reference data relating TBW in Korea. We hope the results of this study will be useful as the baseline data for the evaluation of hydration and nutritional status in healthy and ill adults, including end-stage renal disease (ESRD). PMID- 12089747 TI - Effects of MK-801 and morphine on spinal C-Fos expression during the development of neuropathic pain. AB - The purpose of this study was to investigate the expression of c-fos in the spinal cord during the development of allodynia, induced by peripheral nerve injury. Following tight ligation of the left L5 and L6 spinal nerves of Sprague- Dawley rat, the lumbar spinal cord was postfixed following perfusion. Frontal frozen sections of 40 microm were immunostained according to the peroxidase antiperoxidase method. The allodynic threshold was checked with 8 calibrated von Frey filaments. MK-801 (0.3 mg/kg), morphine (3 mg/kg) and saline (as a placebo) were administered subcutaneously 30 min before, and 24 and 48 hrs after surgery. The tactile threshold decreased below 3 g since 2 days after surgery in the saline and morphine groups, but delayed a little in the MK-801 group. In the superficial layer the number of Fos-like immunoreactive neurones (Fos-LI) peaked at 2 hours and decreased thereafter, and reached normal levels 24 hrs following operation, for all groups. In the deep layer they were biphasic, - peaking at 2 and 24 hrs - in the saline group, but were suppressed in the morphine and MK-801 groups until 7 days following operation. The above discrepancy between the number of Fos-LI and the allodynic threshold showed that central sensitizations are not critically involved in the development of nerve injury induced tactile allodynia. PMID- 12089748 TI - An unusual case of osteoblastic metastasis from gastric carcinoma. AB - We report an unusual case of osteoblastic metastasis from gastric carcinoma. In this case, bone metastasis was the initial manifestation of the cancer. The laboratory findings revealed mild hypocalcemia and markedly elevated alkaline phosphatase levels. Plain X-ray showed mottled osteoblastic changes in the pelvis. Bone marrow and bone biopsy of the pelvis revealed metastatic adenocarcinoma with increased osteoblastic activity. An extensive search for the primary site revealed advanced gastric carcinoma, which was confirmed by endoscopic biopsy. PMID- 12089749 TI - Primary papillary carcinoma arising in a thyroglossal duct cyst. AB - We report a case of papillary carcinoma arising in a thyroglossal duct cyst, presenting with an anterior neck mass of a 31-year-old woman. The tumor was judged to be a primary lesion on the basis of intraoperative examination of the thyroid and pathologic findings of the mass. One year later, a small nodular mass in the left thyroid gland and lymph node enlargement of the right cervical lymph node were noted by follow-up imaging studies. Total thyroidectomy, right modified radical neck dissection and central neck dissection were performed. The thyroid gland revealed nodular hyperplasia without evidence of malignancy. On the other hand, the dissected neck lymph nodes revealed metastatic papillary carcinoma. Taken together, these findings suggested the tumor was a primary papillary carcinoma arising in the thyroglossal duct cyst. PMID- 12089750 TI - A case of primary bilateral adrenal lymphoma (PAL) with central nervous system (CNS) involvement. AB - Primary adrenal lymphoma (PAL) is a very rare disease and usually does not have disease elsewhere, and if present, it is frequently extranodal. Several cases of PALs, with central nervous system (CNS) involvement, gastrointestinal tract or other endocrine organs involvement, have been reported in Western literature. We experienced a case of PAL with CNS involvement, which was previously unreported in Korea. The patient, a 61 year-old male, was admitted with left abdominal pain. After imaging study and needle aspiration biopsy (NAB), a bilateral primary adrenal lymphoma (large B-cell type), with bilateral hemorrhage, was confirmed. Combination chemotherapy, with CEOP regimen, was used. A follow-up abdomino- pelvic CT scan, after the second CEOP chemotherapy, showed the previous adrenal masses had nearly disappeared. However, about 2 months after diagnosis (day 52), he showed recent memory loss, obtundation, disorientation and drowsy mental status. The brain MRI revealed multifocal scattered lesions with increased signal intensity. The result of a brain biopsy was diffuse large B-cell lymphoma. After 5(1/2) cycles of chemotherapy, with CEOP regimen, and whole brain radiotherapy, he is still surviving 6 months later, and has become alert with nearly normalized cognitive function. PMID- 12089751 TI - Ceftriaxone induced immune hemolytic anemia: detection of drug-dependent antibody by ex-vivo antigen in urine. AB - There have been a few reported cases of immune hemolytic anemia induced by ceftriaxone. We encountered a patient with immune hemolytic anemia that seemed to be stimulated by a degradation product of ceftriaxone. The patient's direct antiglobulin test was positive only for C3d, and no ceftriaxone-dependent antibodies were detectable in the patient's serum. To demonstrate the presence of the ceftriaxone-induced antibodies, an ex-vivo antigen in urine was obtained from the patient. In addition, we prepared a 1 mg/mL suspension solution of ceftriaxone, and group AB serum as a complement source. Using several combinations of the above reactants, the indirect antiglobulin test was performed. Only the indirect antiglobulin test using the patient's serum with the ex-vivo urine antigen was found to be positive. Other combinations were not reactive. To our knowledge, this is the first reported case in Korea, in which the causative antibody appeared to be stimulated solely by a degradation product of ceftriaxone. PMID- 12089752 TI - Diffuse glomerular basement membrane lamellation in post-transplant IgA nephropathy. AB - Diffuse glomerular basement membrane (GBM) lamellation, reminiscent of Alport's syndrome, has rarely, and exclusively, been reported in renal allografts from pediatric donors to adult recipients. We report on a similar lesion, identified in a 42-year-old male, who received a kidney from an unrelated 21-year-old living male donor. The disease of the recipient was unknown. Renal allograft biopsies were performed 3.5 and 4.8 years after the renal transplantation, due to massive proteinuria and serum creatinine elevation. The histological features of both biopsies were similar, but more advanced in the second biopsy. Glomerular mesangium was widened and had an IgA deposit in the first biopsy. In addition to the presence of mesangial electron dense deposits, the GBM showed diffuse lamellation and splintering on the subepithelial side, but no definite deposits. In the second biopsy, IgA deposits were extended to the peripheral capillary walls, but electron microscopic examination was not available. Two months after the second biopsy, the patient returned for hemodialysis. PMID- 12089753 TI - Primary malignant fibrous histiocytoma of the kidney. AB - Primary renal malignant fibrous histiocytoma is a rare tumor of the kidney. It is clinically and radiologically indistinguishable from a renal cell carcinoma. Even following radical surgery, the tumor shows a strong predilection for local recurrence and the prognosis is generally poor. We report on a 32-year-old man with malignant fibrous histiocytoma of the kidney who remained free of recurrence for 1 year after radical nephrectomy. PMID- 12089754 TI - Using a new condensable composite for posterior restorations. AB - This article discusses the properties of condensable composites and how their filler particles are continually being improved to create an updated, advanced tooth-colored material that can be packed with the same clinical success as amalgam in posterior restorations. Many dental companies are now addressing the major concerns of shrinkage, polishability, and discoloration that inundated previous composites by introducing the next generation of condensable composites. With increased filler content and the addition of special additives to the resin phase, Prodigy Condensable allows clinicians to follow the same protocol for placing amalgam restorations while being assured of improved wear-resistance, strength, longevity, postoperative sensitivity, and esthetics. PMID- 12089755 TI - An advance in condensable composites. AB - With the constant advance of technology and the public's increasing awareness of esthetically pleasing restorations, alternatives for amalgam and early composite materials have been brought to the forefront of dentistry. Condensable composites offer characteristics that distinguish them from their traditional hybrid counterparts: decreased wear, packability, and bulk curing with less polymerization shrinkage. Prodigy Condensable expands the first generation of condensables by altering its resin phase to make the material more packable. Through increased filler content and methacryoyloxyethlpolycaprolactonephosphate and rheological control additives, this second-generation condensable composite possesses strength and ease of use comparable to and even exceeding amalgam while demonstrating superior esthetics for all posterior restorations. PMID- 12089756 TI - Condensable composites: the next generation. AB - The search for a tooth-colored restorative material with the physical and handling characteristics of amalgam has been ongoing for many years. The esthetic awareness of our patients is such that many insist on tooth-colored replacements when they need a dental restoration. The introduction of condensable composites is the latest step toward that goal. This article will discuss the evolution of this restorative option, the latest advancements in the technology, and describe in detail the techniques for placement and finishing of this material. The use of esthetic composite tints also will be discussed. PMID- 12089757 TI - Postmenopausal estrogen replacement and tooth retention. AB - Loss of underlying supportive alveolar bone is a major cause of tooth loss. Alveolar bone loss appears to be related to the status of the extracranial skeleton, and therapies that preserve skeletal bone may be expected to benefit tooth retention. An association between postmenopausal hormone replacement therapy (HRT) and tooth retention was found in a cohort of 488 elderly women, 72 to 95 years old, who participated in the Framingham Heart Study between 1948 and 1995. There was an association between duration of HRT and tooth retention for total number of teeth remaining and the individual types of teeth (incisors, canines, and premolars, but not molars) retained. The odds of being edentulous were reduced by 6% for each 1-year increase in duration of HRT use. The data suggest that postmenopausal HRT protects against tooth loss and reduces the risk of edentulism. PMID- 12089758 TI - Is estrogen deficiency a risk factor for periodontal disease? AB - In recent years, interest in the relationship between osteopenia, osteoporosis, estrogen status, and periodontal disease has grown considerably. A substantial number of studies have attempted to address this important question. Despite the large body of evidence, controversy persists concerning a possible association between these conditions. Whether loss of oral bone is related to loss of systemic bone and whether estrogen deficiency modulates loss of oral bone are subjects of more specific debate. This article reviews the direct and indirect scientific evidence supporting the role of estrogen deficiency as a risk factor for periodontal disease. A model is proposed for the biological basis of such increased risk. The article also suggests essential criteria for the design of studies examining the relationship between osteopenia, osteoporosis, estrogen deficiency, and periodontal disease. PMID- 12089759 TI - Effect of estrogen supplementation on periodontal disease. AB - This article presents evidence of the protective role of estrogen supplementation on periodontal disease in postmenopausal women. A total of 412 women, 236 postmenopausal (50 to 75 years old) and 176 premenopausal (25 to 49 years old), were included in this study. The postmenopausal group included 59 women receiving estrogen replacement therapy (ERT) and 177 who did not receive ERT (non-ERT). Periodontal disease was assessed from two independent but highly correlated measures: clinical attachment loss (CAL) and alveolar bone loss (ABL). Severe periodontal disease was arbitrarily defined as mean CAL or ABL > or = 3 mm. Data adjusted for age effect were compared for various groups of premenopausal and postmenopausal women. Non-ERT women were twice as likely as their ERT counterparts, and three times more likely than premenopausal women, to exhibit severe attachment loss. Individual percentages of women affected by severe attachment loss were 18.6%, 11.9%, and 6.3% for non-ERT, ERT, and premenopausal women, respectively. On the other hand, severe ABL was detected in 34%, 20.3%, and 9.7% of the non-ERT, postmenopausal ERT, and premenopausal women, respectively. Similar to CAL, when ABL was used as a measure of disease, almost twice as many non-ERT women compared to their ERT counterparts and four times compared to premenopausal women exhibited severe ABL. Therefore, ERT appears to have a protective effect on the severity of periodontal disease and the periodontium. PMID- 12089760 TI - Estrogen replacement therapy--something to smile about. AB - Tooth loss and alveolar residual ridge resorption are significant oral health problems in older adults. Although the causes of tooth loss are many, several studies show an association between tooth loss/residual resorption and systemic osteoporosis. Because estrogen replacement therapy (ERT) slows the development of osteoporosis, we investigated the relationship between estrogen use and tooth loss among elderly women who were participating in the Leisure World Cohort Study. Among 3,921 women 52 to 109 years old (median = 81), those who had ever used estrogen retained more teeth than nonusers (21.2 vs 19.2). The age-adjusted risk of having fewer than 25 teeth decreased with increasing duration of ERT: 0.87 for < 4 years of ERT, 0.74 for 4 to 14 years, and 0.70 for 15+ years compared to nonusers. Similarly, the risk of edentia was inversely related to estrogen use. If a woman had ever taken ERT, her risk of losing all her teeth was about two thirds that of a woman who had never used estrogen. Long-term users (15+ years) had half the risk of becoming toothless. Despite differences in age, education, methods of obtaining tooth and hormone data, and prevalence of tooth loss, two other cohort studies (the Nurses' Health Study and the Framingham Heart Study) reported similar reductions in tooth loss among estrogen users. The data suggest that tooth retention, in addition to being affected by osteoporosis of alveolar bone, also shares in the benefits of ERT on osteoporosis. PMID- 12089761 TI - Tooth loss and hormone use in postmenopausal women. AB - Nearly 32% of US women aged 65 to 69 have no teeth. Because some tooth loss may reflect systemic osteoporosis, and postmenopausal hormone therapy protects against osteoporosis, the authors examined the risk of tooth loss in relation to hormone use in a prospective study of 42,171 postmenopausal women. Overall, the risk of tooth loss was 24% lower in women who were currently using hormones, although there was little effect for women who had stopped taking hormones. This decreased risk for current hormone users was observed regardless of the duration of use and was similar for a variety of hormone preparations and doses. Although few studies have examined the issue, this research suggests that hormone therapy may reduce tooth loss. PMID- 12089762 TI - Long-life physical property preservation and postendodontic rehabilitation with the Composipost. AB - Most coronal radicular reconstructions are made of cast inlay core metals or prefabricated posts covered in composite. The differences in the mechanical properties of these elements create a heterogeneous mass with inconsistent mechanical behavior. Studies using the Finite Element Method have shown the biomechanical disturbances caused by the inclusion of materials with a modulus of elasticity that is superior to that of dentine (i.e., nickel, chrome, zircon, etc). The use of materials with a modulus of elasticity close to that of dentine does not disturb the flow of stress inside the root. To our knowledge, only a composite material structured with programmable mechanical properties would be capable of producing both high mechanical performance and a modulus of elasticity adapted to dentine values. The C-POST, made of carbon epoxy, accommodates the demands of the dentine, as well as the in vitro stress linked to the prosthesis. The internal structure, consisting of long high-performance carbon fibers, unidirectionally and equally stretched, confers a totally original behavior that is adapted to clinical objectives. In addition, the C-POST has a fracture resistance superior to most metals. PMID- 12089763 TI - Six years of in vitro/in vivo experience with Composipost. AB - Some years ago, we created the following classifications for endodontic posts: (1) first generation posts (self-threaded posts, screw posts, serrated-carved posts); (2) second generation posts (passive posts); (3) third generation posts (nonmetallic passive posts). In this last group, we the placed carbon-fiber posts, which have a modulus of elasticity very similar to the modulus of elasticity of dentine and can realize a tooth-post-core monobloc instead of an assemblage of heterogeneous materials. This is quite a new philosophy in rebuilding endodontically treated teeth and is based on the use of integratable materials that homogeneously distribute masticatory loads and reduce stress. PMID- 12089764 TI - Evaluation of post and cores in the laboratory: rationale for developing a fatigue test and preliminary results. AB - Prosthetic treatment failures related to the biomechanical deficiencies of post and cores still represent a problem of clinical significance. To overcome the difficulties of clinical studies, numerous in vitro methods were developed to address specific properties of post-and-core restorations. Most of them, however, were based on an oversimplified mechanical testing of the restored tooth. Experience proved that the fatigue of restorative materials was a primordial factor in clinical failures. Therefore, special devices were built that stimulate the physiological masticatory cycle. Tests performed with adhesive post-and-core systems seem to indicate that materials placed in the tooth should have physical properties as close as possible to those of natural tissues. New carbon-epoxy posts appear to offer a promising solution for restoring the endodontically treated tooth. PMID- 12089765 TI - Coronoradicular rehabilitation with a carbon-fiber post. AB - Endodontically treated teeth present a myriad of problems because of coronal destruction from caries, previous restorations, and coronal access. Previously, a post was considered necessary to strengthen a tooth and to resist intraoral forces. However, recent research has indicated that not all endodontically treated teeth require a post, core, and crown. Each clinical situation must be evaluated and the need for a post and/or the appropriate post determined. The recent introduction of a carbon-fiber post may reduce some of the problems inherent with metal posts. Proper placement, advantages, and disadvantages are discussed. PMID- 12089766 TI - The C-POST system. AB - This article represents a clinical report from daily experience with the C-POST system, a carbon-fiber post for core build-up. The C-POST was invented in France in 1988 and introduced in Sweden in 1992. When I first started using the system, little literature about it had been published. Because the C-POST fits well in the modern bonding concept, it has become my therapy of choice. In my practice, more than 900 treated teeth have been registered in a computer so that treatment progress can be followed. Every treated tooth was registered according to the type of primer, cement, core material, number of posts, and type of prostheses used. The prosthetics included large composite fillings, porcelain crowns, and/or metal crowns or bridges from 3 to 12 units. Of the 900 treated teeth, 173 of them are between 3 and 4 years old, 2 contain fractured roots, and 1 had a post fracture. This article describes how to fabricate a C-POST core build-up and contains clinical hints and advice. Other applications for the post, such as splinting, bridges, and repair of fixed prosthodontics and vital teeth, are discussed. PMID- 12089767 TI - Carbon-fiber post removal technique. PMID- 12089768 TI - A history of pepsin and related enzymes. AB - Studies on gastric digestion during 1820-1840 led to the discovery of pepsin as the agent which, in the presence of stomach acid, causes the dissolution of nutrients such as meat or coagulated egg white. Soon afterward it was shown that these protein nutrients were cleaved by pepsin to diffusible products named peptones. Efforts to isolate and purify pepsin were spurred by its widespread adoption for the treatment of digestive disorders, and highly active preparations were available by the end of the nineteenth century. There was uncertainty, however, as to the chemical nature of pepsin, for some preparations exhibited the properties of proteins while other preparations failed to do so. The question was not settled until after 1930, when Northrop crystallized swine pepsin and provided convincing evidence for its identity as a protein. The availability of this purified pepsin during the 1930s also led to the discovery of the first synthetic peptide substrates for pepsin, thus providing needed evidence for the peptide structure of native proteins, a matter of debate at that time. After 1945, with the introduction of new separation methods, notably chromatography and electrophoresis, and the availability of specific proteinases, the amino acid sequences of many proteins, including pepsin and its precursor pepsinogen, were determined. Moreover, treatment of pepsin with chemical reagents indicated the participation in the catalytic mechanism of two aspartyl units widely separated in the linear sequence. Studies on the kinetics of pepsin action on long chain synthetic peptides suggested that the catalytic site was an extended structure. Similar properties were found for other "aspartyl proteinases," such as chymosin (used in cheese making), some intracellular proteinases (cathepsins), and plant proteinases. After 1975, the three-dimensional structures of pepsin and many of its relatives were determined by means of x-ray diffraction techniques, greatly extending our insight into the mechanism of the catalytic action of these enzymes. That knowledge has led to the design of new inhibitors of aspartyl proteinases, which are participants in the maturation of human immunodeficiency virus and in the generation of Alzheimer's disease. PMID- 12089769 TI - Beyond dominance: the importance of leverage. AB - The dominance concept as it is currently defined and applied in animal behavior is problematic. What has been traditionally considered dominance is actually a combination of dominance in the strict sense and power based upon other sources. Rather than working within the current paradigm, I propose a conceptual revision founded upon the more inclusive concept of power. Power is a phenomenon where a dyadic relationship is asymmetrical (Simon 1953) and can be divided into two types: dominance and leverage. Dominance is power based upon the ability to use force. Leverage is power based upon a resource that cannot be taken by force. Four characteristics of power are used in sociology (base, means, amount, and scope) that facilitate both the expansion of the power concept beyond traditional dominance and the application of these theoretical ideas in empirical studies. This cross-disciplinary approach to power allows a wide range of behaviors to be considered as critical while at the same time it focuses the attention of researchers to the aspects of power that differ among dyads, classes, and species. Power is not simply a linear combination of dominance and leverage, and more research is needed before the exact nature of this relationship can be clarified. By considering dominance as one form of power, this framework fosters a more complete understanding of power dynamics and their effects on animal societies. PMID- 12089770 TI - Where is the Thomas Henry Huxley of environmentalists? PMID- 12089772 TI - Scope of perioperative nursing practice. PMID- 12089771 TI - Managing the angry, hostile, finger pointing individual with success. PMID- 12089773 TI - "Just say no" to abuse in the workplace. PMID- 12089774 TI - Oh my, the pressure! AB - The intraoperative phase of a surgical patient's hospital stay has been overlooked as a major contributor of pressure ulcers that may arise postoperatively. Pressure ulcers are defined and then the hazards, underlying knowledge, and prevention tactics are reviewed. Bed sore, decubitous ulcer, pressure sore, and pressure ulcer are different terms describing the same problem encountered by medical and surgical patients. The common denominator is pressure- sustained pressure. PMID- 12089775 TI - Filicide: a review. PMID- 12089777 TI - The prevalence of extralegal deprivation of liberty in a psychiatric hospital population. PMID- 12089776 TI - "New" public management of mentally disordered offenders: Part I. A cautionary tale. PMID- 12089778 TI - Telephone scatologia. Comorbidity with other paraphilias and paraphilia-related disorders. PMID- 12089779 TI - Prediction of dangerousness in mentally disordered offenders in Germany. PMID- 12089780 TI - Gender and fitness to stand trial. A 5-year review of remands in Quebec. PMID- 12089781 TI - Predictive validity of the Violence Risk Appraisal Guide: a tool for assessing violent offender's recidivism. PMID- 12089782 TI - [Anthrax. The growing danger]. PMID- 12089783 TI - [Molecular pathologic diagnosis in solid tumors. What is clinically relevant?]. AB - There are only a few clinically relevant applications of molecular pathology assays in solid tumors. Among the findings which may influence therapy decisions are the amplification of Her-2/new in breast cancer and specific translocations in sarcomas. Mutation analyses of p53 may be helpful for only a very few cases, e.g. for confirming high grade dysplasia in the upper gastrointestinal tract or for diagnosing malignant soft tissue tumors in isolated cases. Microsatellite analyses are important for HNPCC screening or distinguishing tissue specimens of questionable identity. Other applications of molecular pathology assays such as detection of minimal residual disease or tumor cell dissemination and FISH analysis of urine and effusion specimens, may be increasingly applied in the future. PMID- 12089784 TI - [Molecular diagnosis in hematopathology]. AB - The diagnosis of hematopoietic neoplasias has nowadays become more and more refined by molecular cytogenetic and molecular biological techniques. The biological basis of most of these techniques is that (malignant) tumors represent clonal neoplasias that are derived from a single or few progenitor cells, thus imparting a clonal relationship to all daughter cells. Apart from clonality analysis as a marker for neoplastic growth, the presence or absence of certain primary chromosome abnormalities, and the number and the kind of secondary genetic anomalies become increasingly important in the delineation of the biological grade of aggressiveness. PMID- 12089785 TI - [Telomerase and new aspects for tumor biology]. AB - The field of tumor genetics is characterized by a whole string of unsolved questions that affect both the basics of tumorigenesis and the possibilities of using molecular markers at the clinical level. Telomerase, an enzyme that has the capability of unlimitedly maintaining the division of immortal cells, can nowadays be demonstrated with the use of up-to-date molecular biological methods. In vitro experiments have now succeeded in changing normal cells into tumor cells via transfection with telomerase. The regulatory mechanisms and interactions of telomerase are presently the subject of intensive research. Telomerase activity plays a crucial role even in present day diagnostic procedures and the estimation of prognosis for certain tumor types. In comparison with normal tissues, increased telomerase activity in tumor tissues implies therapeutical possibilities with the use of telomerase inhibitors. PMID- 12089786 TI - [Immunohistochemical diagnosis in cancer metastasis of unknown primary tumor]. AB - Immunohistochemical studies on metastatic carcinomas of unknown primary site are cost-effective and often allow a specific identification of the tumour origin, especially if the metastases are adenocarcinomas by light microscopy. Commercially available site-specific markers include prostate-specific antigen, thyroglobulin, thyroid transcription factor-1, uroplakin III, GCDFP-15, oestrogen and progesterone receptors, alpha-fetoprotein, the A103 monoclonal antibody against MART-1, cytokeratins 7 and 20, cytokeratins of basal cell type, p63, carcinoembryonic antigen, CA125, EMA, vimentin, HepPar-1, WT-1 and S100 protein. However, immunostaining with most of these markers does not show an absolute specificity for a certain primary site. For this reason, histopathologists interpretating staining results with these markers should take the available clinical data and the histological features of the metastatic carcinoma into consideration. These data are necessary to estimate the relative a priori probability of possible carcinomas. Based on Bayes' theorem, the a priori probability can then be used to calculate the diagnostically relevant predictive values for immunostaining results with the chosen markers. PMID- 12089787 TI - [The future of telepathology. An Internet "distributed system" with "open standards"]. AB - With the availability of Internet, the interest in the possibilities of telepathology has increased considerably. In the foreground is thereby the need of the non-expert to bring in the opinions of experts on morphological findings by means of a fast and simple procedure. The new telepathology system iPath is in compliance with these needs. The system is based on small, but when possible independently working modules. This concept allows a simple adaptation of the system to the individual environment of the user (e.g. for different cameras, frame-grabbers, microscope steering tables etc.) and for individual needs. iPath has been in use for 6 months with various working groups. In telepathology a distinction is made between "passive" and "active" consultations but for both forms a non-expert brings in the opinion of an expert. In an active consultation both are in direct connection with each other (orally or via a chat-function), this is however not the case with a passive consultation. An active consultation can include the interactive discussion of the expert with the non-expert on images in an image database or the direct interpretation of images from a microscope by the expert. Four software modules are available for a free and as fast as possible application: (1) the module "Microscope control", (2) the module "Connector" (insertion of images directly from the microscope without a motorized microscope), (3) the module "Client-application" via the web-browser and (4) the module "Server" with a database. The server is placed in the internet and not behind a firewall. The server permanently receives information from the periphery and returns the information to the periphery on request. The only thing which the expert, the non-expert and the microscope have to know is how contact can made with the server. PMID- 12089788 TI - [WHO classification of Hodgkin's lymphoma and its molecular pathological relevance]. AB - The current WHO classification of Hodgkin's lymphoma (HL) generally distinguishes the relatively rare variant (approximately 5% of all cases of HL) of nodular lymphocyte predominant type from a second group, which comprises classical HL and is separated into four subtypes: lymphocyte rich type, nodular sclerosis type, mixed cellularity type and lymphocyte depleted type. The classical lymphocyte rich subtype is a new entity and based on the typical morphology, can be recognized by definition only by the immunohistochemical characteristics of the Hodgkin and Reed/Sternberg cells (HRS) (CD30+, CD15+, CD20-). Molecular single cell studies are consistent with the dichotomy of HL in nodular lymphocyte predominant and classical types and stress the exceptional position of the former, which shows similarities with non-Hodgkin's lymphomas in several aspects. On the molecular biology level the tumor cells of all kinds of HL turn out to be clonal B cells derived from germinal center cells. However, tumor cells of nodular lymphocyte predominant HL differ from those of classical HL by the pattern of somatic mutations. Considering the inability of HRS cells of classical HL to express a B cell receptor, they should perish under normal conditions. However, they escape from apoptosis by mechanisms so far only partially understood, such as genomic mutations of the l-kappa B gene and the fas receptor gene or probably by down-regulation of B cell markers. In rare cases, HRS cells of HL can also be derived from T cells, as could be demonstrated by single cell analysis. Also, it could be shown by single cell PCR that HL and non-Hodgkin's lymphoma of both B and T cell types can arise from a common precursor. These results suggest that future classifications of HL will not only take into account the morphological and phenotypical profile, but also mechanisms of transformation yet to be discovered. PMID- 12089789 TI - [Morphological diagnosis of spinal diseases. Current technical possibilities and challenges for the histological preparation of transpedicular biopsies]. AB - The spine is the central component for the mobility of the human body. Both locally limited and diffuse pathologies of the spine are a challenge for the treating physician due to the difficult anatomy. The biopsy of a pathologically altered vertebral body is a fast and reliable basis for further therapy but until now this has not regularly been made use of as a diagnostic standard for spinal diseases, since the tissue gained was often microfractured because of the difficult anatomical position. Our own experience with transpedicular vertebral biopsies of 70 patients with different diseases is reported because of the considerable improvement in the biopsy technique and the methodical possibilities for processing the bony tissue. Methods which have proven particularly valuable are contact radiographs, embedding in plastic, careful decalcifying with EDTA and immunohistological procedures. In this way a definite diagnosis can be made in 97% of the cases. A close cooperation with the clinician carrying out the biopsy and a greater use of methods other than just fast decalcification is recommended. PMID- 12089790 TI - [Fatal cardiomyopathy in adult in polyglucosan body disease]. AB - Adult polyglucosan body disease (APBD) is a rare genetic disorder, inherited in an autosomal recessive mode. The disease is caused by mutations of the gene coding for the glycogen-branching enzyme, which is essential for branching of polyglucose chains in the normal glycogen molecule. The age of clinical manifestation of the disease mostly is between 40 and 60 years and its course is slowly progressive. Characteristic globular deposits (polyglucosan bodies, PGB) can be detected in biopsies of skin and skeletal muscle as well as in the peripheral and central nervous system. Biochemically, PGBs consist of poorly branched glycogen molecules with abnormally long polysaccharide chains. We report the case of a 50-year-old female patient with APBD who suffered from neurological symptoms such as spastic tetraparesis, urinary incontinence, hypesthesia and dementia. She died unexpectedly of cardiac failure. At autopsy a severe cardiomyopathy with abundant PGBs in the heart muscle fibres could be proven as the cause of death. This observation shows that in addition to the known deposition of PGBs in nervous system and skeletal muscle, an involvement of the heart has to be considered in APBD as well. PMID- 12089791 TI - [Instrumentation and implant systems for the spine]. PMID- 12089792 TI - [Effect of artificial disk nucleus implant on mobility and intervertebral disk high of an L4/5 segment after nucleotomy]. AB - This study investigated whether after a nucleotomy and implantation of a prosthetic disk nucleus (PDN) the original height and mobility of an L4/L5 disk can be restored. Compared to the intact state (100%), nucleotomy increased the median values of the normalized range of motion (ROM) in flexion/extension to 118%, lateral bending to 112%, and axial rotation to 121%. PDN implantation reduced ROM to 102%, 88%, and 90%. These differences were even more distinct when comparing the neutral zone (NZ) with 210%, 173%, and 107% after nucleotomy and 146%, 149%, and 44% after PDN implantation. With an axial preload of 200 N, disk height after nucleotomy was reduced by about 1.3 mm and could be restored with PDN implantation. PDN implantation can restore disk height and ROM after nucleotomy to normal values and reduce the strong NZ increase. Further biomechanical characterization of this therapy with PDN is necessary. PMID- 12089793 TI - [Basic principles of successful implantation of the SB Charite model LINK intervertebral disk endoprosthesis]. AB - The scope of the LINK SB Charite Artificial Disc is to replace degeneratively changed, symptomatic intervertebral discs and to restitute the fundamental functions of the lumbar mobile segment. Biomechanical static- and dynamic test results as well as postoperative controls of a patient-group with uniform indications are presented. The options of an intervertebral disc replacement with the SB Charite Artificial Disc are discussed. For the evaluation of segmental stability, comparative static examinations of the range of motion of cadaveric lumbar segments prior to and after the implantation of the artificial disc were carried out. The stress-dependent mechanical response to directional static load on implanted SB Charite Discs in neutral position, extension and flexion was examined under different testing-conditions in order to gain results on changes of surfaces, durability and sustain of functionality. Restitutable changes to the UHMWPE sliding core, prognosed definite deformations of the sliding core after 5 and 10 years, and the functionality of the UHMWPE sliding cores after stress were examined using dynamic compression tests. Our clinical results refer to 20 patients, who received a mono- (n = 14) or bisegmental (n = 6) implantation of a SB Charite Artificial Disc following a postnucleotomy-syndrome. Comparison of the ranges of motion in cadaveric segments with and without a SB Charite Artificial Disc reflected a significantly higher mobility in cases with an implanted prosthesis only in axial rotation. Other movements showed similar ranges of motion in native segments and in segments with prosthesis. The UHMWPE sliding cores showed tolerable changes during maximal static compression in neutral position and extension. The 7.5 mm sliding cores showed changes beyond normal deformations on compression in maximal flexion. None the less it may be concluded that in vivo these sliding cores also retain their mechanical integrity. Most of the 7.5 mm and 9.5 mm high sliding cores showed a regeneration of 50% compared to maximal deformation. All the tested UHMWPE sliding cores reflected continued articulation despite plastic deformation. Predictions for a period of 5 and 10 years state that extreme loads of 4.5 kN would lead to a reduction in the height of the sliding cores of less than 8%. There was no failure of the sliding cores due to delamination or extreme coldflow. The Oswestry-Score amounted to 24% (0 56) in the postoperative controls 46 months (6 months-13 years) postoperatively. In pain analysis 5 patients (25%) were pain free, 14 patients (70%) reported a pain reduction and 1 patient (5%) had no pain relief compared to preoperatively. 15 (83%) of 18 patients had no leg pain anymore after implantation of the SB Charite Artificial Disc. 14 (93%) of 15 patients had a negative leg raising test. 2 of 5 patients recovered from their motor symptoms. The segmental saggital total range of motion was 6.8 degrees (0-20) and in cases of bisegmental implantation 5.8 degrees (0-14) in the second disc segment. Those neighboring segments with the best function showed a range of motion of 6.8 degrees (2-15). The stated general and specific complications (n = 9) had a definite influence on the results of the postoperative control in one patient and a questionable influence in another patient. Based on biomechanical static and dynamic testing, the SB Charite Artificial Disc is suited to replace the fundamental functions of the lumbar intervertebral disc. Patients suffering from a postnucleotomy-syndrome can be rendered pain free with the implantation of this prosthesis. PMID- 12089794 TI - [Fusion implants of carbon fiber reinforced plastic]. AB - Carbon fiber reinforced plastics (CFRP) are used in the medical field when high mechanical strength, innovative design, and radiolucency (see spinal fusion implants) are needed. During the manufacturing process of the material CFRP carbon fibers are embedded into a resin matrix. This resin material could be thermoset (e.g., epoxy resin EPN/DDS) or thermoplastic (e.g., PEAK). CFRP is biocompatible, radiolucent, and has higher mechanical capabilities compared to other implant materials. This publication demonstrates the manufacturing process of fusion implants made of a thermoset matrix system using a fiber winding process. The material has been used clinically since 1994 for fusion implants of the cervical and lumbar spine. The results of the fusion systems CORNERSTONE-SR C (cervical) and UNION (lumbar) showed no implant-related complications. New implant systems made of this CFRP material are under investigation and are presented. PMID- 12089796 TI - [Effect of design and implantation technique on risk of progressive sintering of various cervical vertebrae cages]. AB - The aim of this study was to compare the subsidence of differently designed cervical interbody fusion devices under defined conditions. Forty-five bovine vertebral bodies were dissected from soft tissue and cartilage. The bony end plate was then taken off by 0, 1, and 2 mm. Five vertebral bodies of each abrasion depth were prepared for the uptake of a fusion device. Thus, three different fusion devices of comparable size underwent biomechanic testing in a Zwick testing machine with 4000 cycles of axial compression between 50 and 1000 N. Every 1000 cycles, the subsidence into the vertebral body was measured. Abrasion of the end plate resulted in an increased subsidence. The cage with rectangular shape and the cage with cylindric body and lateral wings showed better resistance to axial compression as long as the end plate remained intact. When the end plate was taken off, the subsidence was as high as in the cylindric cage, of which the subsidence did not correlate to the end plate abrasion. During preparation of the implant bed, the cortical bone of the end plate must be treated carefully. In cases of intact end plate, rectangular supporting areas can decrease the risk of subsidence. PMID- 12089795 TI - [Mechanical studies of lumbar interbody fusion implants]. AB - In addition to autogenous or allogeneic bone grafts, fusion cages composed of metal or plastic are being used increasingly as spacers for interbody fusion of spinal segments. The goal of this study was the mechanical testing of carbon fiber reinforced plastic (CFRP) fusion cages used for anterior lumbar interbody fusion. With a special testing device according to American Society for Testing and Materials (ASTM) standards, the mechanical properties of the implants were determined under four different loading conditions. The implants (UNION cages, Medtronic Sofamor Danek) provide sufficient axial compression, shear, and torsional strength of the implant body. Ultimate axial compression load of the fins is less than the physiological compression loads at the lumbar spine. Therefore by means of an appropriate surgical technique parallel grooves have to be reamed into the endplates of the vertebral bodies according to the fin geometry. Thereby axial compression forces affect the implants body and the fins are protected from damaging loading. Using a supplementary anterior or posterior instrumentation, in vivo failure of the fins as a result of physiological shear and torsional spinal loads is unlikely. Due to specific complications related to autogenous or allogeneic bone grafts, fusion cages made of metal or carbon fiber reinforced plastic are an important alternative implant in interbody fusion. PMID- 12089798 TI - [Dislocation tendency, stabilizing effect and sintering tendency of different lumbar vertebrae cages in an in vitro experiment]. AB - For biomechanical purposes, interbody fusion cages should not dislocate, should provide high stability, and should have a low subsidence risk. Zientek (Marquardt Medzintechnik), Stryker (Stryker Implants), and Ray lumbar interbody fusion cages (Surgical Dynamics) were tested in this study. They were implanted by pairs from a posterior approach without further stabilization. In a first step, each cage design was implanted into four human L3-4 segments and extracted posteriorly under an axial preload of 200 N. In a second step, standard flexibility tests were carried out with 24 human L2-3 and L4-5 specimens in an intact condition, directly after cage implantation, and after cyclic axial compression loading (200 1000 N, 40,000 cycles, 5 Hz). In a third step, a destructive axial compression test was carried out. Maximum pullout force was highest with Ray cages (median 945 N), followed by Zientek (605 N) and Stryker cages (130 N). With all three cage designs, primary stability was higher in lateral bending and flexion than in extension and axial rotation. Implantation of Ray cages caused a decreased range of motion in all three loading directions ranging between 49% and 99%. Zientek cages only stabilized in lateral bending, flexion, and extension (45-78%) and Stryker cages in none of the three loading directions. Cyclic loading caused an increased range of motion in all cases up to 190%. Axial compression force at failure was 8413 N with Ray cages, 8359 N with Stryker cages, and 5486 N with Zientek cages. The cage design seems to influence the dislocation tendency. In this regard, threaded cages or cages with anchorage systems seem to provide more security. The stabilizing effect seems to be mainly influenced by factors such as the degree of distraction or destruction of the facet joints rather than by the cage design. PMID- 12089797 TI - [Stabilizing effect and sintering tendency of 3 different cages and bone cement for fusion of cervical vertebrae segments]. AB - Important requirement for spinal fusion devices for segment are that they provide sufficient stability and guarantee a low subsidence risk. An important requirement for spinal fusion devices for segments are that they provide sufficient stability and guarantee a low subsidence risk. Therefore, in the following in vitro study, the stabilizing effect and subsidence tendency of cervical fusion cages and bone cement were investigated during cyclic loading. The WING cages (Medinorm AG) and BAK cages (Spinetec) made of titanium, the carbon fiber reinforced PEEK cage from Acromed (DePuy Acromed), and bone cement (PMMA, Sulzer) were tested. Twenty-four human cervical spine specimens were first tested intact with a standardized flexibility test (+/- 2.5 Nm). Then the implants were inserted and the primary stability determined. For the simulation of the postoperative loading of the cervical spine a cyclic loading protocol with 700 loading cycles was performed. In this test pure moments +/- 2.0 Nm in 9 different loading directions in randomized order were applied together with a 50 N preload to simulate the weight of the head. The subsidence and "long term stability" was measured after 50, 100, 200, 300, 500, and 700 cycles. All implants had a stabilizing effect in all directions most obviously in lateral bending. Here the range of motion was between 20.9% (AcroMed Cage), and 62% (BAK Cage) with respect to the intact specimen (100%). In laterial bending, flexion, and axial rotation the AcroMed cage stabilized the most followed by the bone cement, WING and BAK Cage. In extension the specimens treated with bone cement were the most stable. After 700 loading cycles the specimens with the BAK cage lost 1.6 mm in height, with the WING Cage 0.8 mm, with the Acromed 0.7 mm, and with the bone cement 0.5 mm. Two Acromed Cages dislocated during the long term testing. Cages have the potential to stabilize as effectively as bone cement. A smaller contact area, however, causes a higher subsidence risk compared to bone cement but increases the fusion area, thus increasing the chance of obtaining bony fusion. PMID- 12089799 TI - [In vitro stability study of an improved implant system for minimally invasive transforaminal approach]. AB - Calf specimens from the L3-4 and L5-6 levels were used for in vitro stability testing of a new transforaminal lumbar interbody fusion (TLIF) implant. Results were compared with those of the conventional posterior lumbar interbody fusion (PLIF) technique using two cuboid spacers. The specimens were loaded with pure moments of 10 Nm in flexion, extension, lateral bending, and axial rotation without any axial preload. They were allowed to move freely and unconstrained in all directions. In extension the PLIF implants showed slightly higher degrees of stiffness than the TLIF implant. While the conventional PLIF technique results in an increased range of motion by a factor of 2.5 after implant insertion, the TLIF approach prevents segmental destabilization in axial rotation. The facet joint arthrodesis using resorbable pins reveals biomechanically interesting results and will therefore be investigated in further studies. PMID- 12089800 TI - [The MACS-HMA hollow screw. An alternative possibility for stable implant anchoring in the vertebral body also for long fusions]. AB - Our biomechanical in vitro tests compared the stability of the MACS HMA (Aesculap, Tuttlingen) implants to three established systems. The MACS HMA is a modular system consisting of porous hollow titanium screws with an outer diameter of 12 mm for monocortical use. We report the preliminary results of MACS HMA used for correction of scoliosis. All other implant systems used are designed with bicortical screws; one is for application with two screws/two rods and one uses a pullout-resistant nut behind the opposite cortex to increase stability. Significantly increased motion in the craniocaudal direction was recognized for bicortical standard screws (20 +/- 17 microns) compared to MACS HMA (10 +/- 11 microns) or the dual screw system (12 +/- 6 microns). Two-tailed t-test showed significantly higher stability for the dual screw system (4.2 kN) and the system with pullout-resistant nut (4.0 kN) compared to all other systems (p < 0.025 or higher). Bicortical implants (2.1-3.2 kN) and MACS HMA (2.6 kN) did not reveal significant differences in pullout strength. All biomechanical tests and in vivo use demonstrated favorable performance of MACS HMA implants. PMID- 12089801 TI - [Effect of implants for replacement of a vertebral body on the mechanical function of the lumbar spine]. AB - A three-dimensional, nonlinear finite element model of the lumbar spine was created. A bisegmental internal spinal fixation device and successively two kinds of vertebral body replacements were integrated into the computer model. The model was loaded with pure moments in the three anatomical main planes as well as with forces that are expected during standing. A vertebral body replacement drastically reduces the mobility in the implant region. An implant has only a minor influence on the mobility and stresses in the adjacent regions for the loading cases chosen. A preload, for example, caused by additional distraction from the precisely fitted ventral implant, exerts a very strong effect on the stresses in the end plates that are in contact with the implant. Different sizes of the contact area between implant and vertebral body have only a local effect on stress distribution. PMID- 12089802 TI - [Stability of ventral, dorsal and combined spondylodesis in vertebral body prosthesis implantation]. AB - The purpose of this study was to evaluate the biomechanical characteristics of short-segment anterior, posterior, and combined instrumentations in lumbar spine vertebral body replacement surgery. Eight fresh frozen human cadaveric thoracolumbar spine specimens (T12-L4) were prepared for biomechanical testing. Pure moments (2.5, 5, and 7.5 Nm) of flexion-extension, left-right axial torsion, and left-right lateral bending were applied to the top vertebra in a flexibility machine and the motions of L1 vertebra with respect to L3 were recorded with an optoelectronic motion measurement system after preconditioning. One anterior, two posterior pedicle screw systems, and two combined instrumentations were tested. Load-displacement curves were recorded and neutral zone (NZ) and range of motion (ROM) were determined. The anterior instrumentation, after vertebral body replacement, showed greater motion than the intact spine, especially in axial torsion. Posterior instrumentation provided greater rigidity than the anterior instrumentation, especially in flexion-extension. The combined instrumentation provided superior rigidity in all directions compared to all other instrumentations. PMID- 12089803 TI - [Comparison of different vertebral body prosthesis with reference to migration and primary stability in dorsoventral spondylodesis after corporectomy with and without laminectomy]. AB - In order to allow speedy mobilization after vertebral body replacement, high primary stability as well as prevention of implant dislocation are required. It is unclear whether differences between common vertebral body replacement systems exist and whether spreadable implants improve primary stability. Migration characteristics of 3 different vertebral body replacement systems were determined using human lumbar spine specimens. Primary stability was assessed by loading specimens successively in 5 conditions: native, after dorsal spondylodesis, after laminectomy and after vertebral body replacement. Migration was shown to depend on cage design and spike shape. Dorsal spondylodesis in combination with vertebral body replacement reduced flexion/extension movement effectively independent of cage design. Stabilization potential in lateral bending was limited. None of the cages limited movement under torsional loading effectively. The possibility to spread the cage had a positive effect on stabilization. PMID- 12089804 TI - [Acute pain therapy and management in orthopedics]. PMID- 12089805 TI - [Bone anchored hearing aids always bilateral!]. PMID- 12089807 TI - [Synaptic function in the auditory pathway and genetics of hearing loss]. PMID- 12089806 TI - [Pediatric immunologic aspects of tonsillectomy]. PMID- 12089808 TI - [Differential diagnosis and management of retropharyngeal space-occupying lesions]. AB - BACKGROUND: Diseases of the retropharyngeal space place high request on the diagnostic and therapeutic knowledge of the otorhinolaryngologist. METHODS: Under consideration of six own cases and of all other published case reports, this inhomogeneous symptom is analysed systematically. RESULTS: Commonly, these alterations of the retropharyngeal space are observed incidentally during routine throat examination or in patients suffering from dyspnoea, dysphagia, dysphonia, sleep apnoea syndrome or neck pain. Six own cases show the very inhomogeneous aetiology (retropharyngeal abscess, lipoma, malignant schwannoma, sarcoidosis, aberrant internal carotid artery, Forestier disease). Therefore, before surgery, a sufficient radiologic diagnosis (lateral neck radiography and CT or MRI respectively) is necessary to avoid grave complications (injury to the cervical spine, the brain, the spinal cord or the internal carotid artery). A tissue sample can be obtained by the transoral approach while the lateral cervical approach seems to be the better way to treat a retropharyngeal abscess. CONCLUSION: Because of the very different aetiology of retropharyngeal space diseases the therapy finally depends on the definite diagnosis. PMID- 12089809 TI - [Stereotactic telepresence in otorhinolaryngologic surgery]. AB - BACKGROUND: Intraoperative three-dimensional (3D) navigation systems determine spatial positions and visualize them in radiological data sets. Usually, data from 3-D computed tomography (CT) are used. We have successfully implemented "augmented reality" in routine clinical practice by superimposing positional data and guiding and ancillary structures on the live endoscopic video of the operating site. Thus, optimal access paths and anatomical structures such as the a. carotis interna or the n. opticus can be displayed. METHODS AND RESULTS: With modern telecommunication, any two locations can be connected for intraoperative consultations with a remote expert, and 3D navigation is an ideal complement. We have successfully tested telephone, ISDN, Ethernet, and ATM technology intraoperatively and find that the ARTMA system provides well-developed technology. CONCLUSIONS: Stereotactic telepresence can provide essential aid in difficult surgical procedures, and the ARTMA Knowledge-Guided Surgery system will contribute further to the optimization of computer-assisted surgery. PMID- 12089810 TI - [Spontaneous defects of the lateral cranial base. 1. Clinical aspects, diagnosis and therapy]. AB - BACKGROUND: Spontaneous cerebrospinal fluid otorrhea (SCFO) or rhinorrhea originating from laterobasal defects is uncommon. An immediate closure of the defect is mandatory due to the risk of life-threatening meningitis. The purpose of this paper is to review the literature and to present two additional case reports with emphasis on diagnostic problems and surgical techniques. PATIENTS AND METHODS: Two adults have been treated for SCFO in our clinic. A 62-year-old woman was complaining of pertinent otorrhea after placement of a ventilation tube and conservative treatment of serous otitis for 1 year elsewhere. A 66-year-old male patient had been treated for acute pneumococcal meningitis. He presented with unilateral deafness and vertigo 3 months later. High-Resolution-Computer Tomography (HRCT) revealed a unilateral defect in the female and a bilateral defect in the male patient. A transmastoidal approach was used to amputate the meningoencephalocele and modified (Mini-Craniotomy) in the second (male) patient. The defect was closed in two layers using allogenic material and fibrin glue. RESULTS: One year after surgery both patients were free of symptoms and defect closure remained stable in HRCT and under microscopic view. CONCLUSIONS: SCFO from tegment defects is an uncommon cause of otorrhea and even rhinorrhea. History and clinical presentation of our patients can be regarded as typical for adults with SCFO. Evaluation of history, HRCT and a high index of suspicion is a prerequisite for correct diagnosis. The approach (transmastoidal, transtemporal, Mini-Craniotomy) to repair leaks of the tegmen should be related to the extent of the defect. PMID- 12089811 TI - [Spontaneous defects of the lateral skull base. 2. Etiology and review of the literature]. AB - BACKGROUND: Isolated cases of spontaneous cerebrospinal fluid otorrhea (SCFO) of temporal bone origin with or without meningo(encephalo)cele have been reported. Diagnosis is predicted on a high index of suspicion. Since meningitis may occur secondary to SCFO, optimal therapy with surgical closure of the defect should immediately be performed. This study was undertaken to evaluate the incidence, characteristics and type of SCFO in literature. MATERIALS AND METHODS: We reviewed the reports on SCFO in the world literature since the first description in 1897 by Escat. Clinical presentation, gender, age and surgical management were aspects of our classification. Not included were patients with cerebrospinal fluid leak following any kind of previous temporal bone surgery, head trauma or rupture of the round window membrane. RESULTS: A total of 241 CSFO in 236 patients were found to accomplish the search criteria. Gender was not found to be a risk factor. Two subtypes of SCFO exist: the majority (161 cases) developing through the tegmen tympani and a smaller number (80 cases) with a translabyrinthine SCFO. The latter group can be regarded as the childhood-type with congenital defects of the otic capsule (Mondini dysplasia) presenting with meningitis in 85% of all cases. Adulthood-type SCFO are typically located in the tegmen tympani area presenting with conductive hearing loss or otorrhea following tube placement in 86% of all cases. Recent literature recommends a combined (transmastoidal and middle fossa) approach for closure of defects in the tegmen. CONCLUSIONS: Copious otorrhea through a perforated tympanic membrane or following placement of a ventilation tube, conductive hearing loss or meningitis without impairment of cochlear or vestibular function should be regarded as warning signs for an existing SCFO in adults. Children usually present with symptoms of inner ear anomalies, meningitis and loss of cochlear-vestibular function. High resolution computer tomography is the diagnostic procedure of choice. A follow-up of several years to disclose any recurrence of SCFO or occurrence at a different area should be considered in any patient. PMID- 12089812 TI - [Atomic force microscope (AFM). A nanomanipulator for biophysical studies of stereocilia of the cochlear hair cells]. AB - OBJECTIVES: Studies of the mechanoelectrical sensor system of the hair cell bundle in the cochlea require a manipulation device that enables controlled force application and movement of individual stereocilia in the nanometer range. METHODS: In our atomic force microscope (AFM) setup, the scan is directly controlled in an upright differential interference contrast (DIC) infrared video microscope with a water immersion objective and in the measured AFM image. Here we present studies on hair cells of the mammalian cochlea. RESULTS AND CONCLUSIONS: The resulting images revealed the tips of individual stereocilia of living sensory cells of the organ of Corti and the typical shape of the ciliary bundle. Scanning electron-microscopic (SEM) images of the identical hair bundles obtained after AFM investigation demonstrated that up to four AFM manipulations on the same cell did not cause obvious damage to the surface morphology of the stereocilia. PMID- 12089813 TI - [Laser tonsillectomy in tonsillar hyperplasia of early childhood]. AB - Tonsillotomy, the partial resection of hyperplastic tonsils, is generally rejected in Germany as a treatment option for obstructions of the isthmus faucium in early childhood. We think this approach merits consideration. The favorable results obtained with tonsillotomy in the University Hospital Benjamin Franklin (n = 637), a private clinic, and an ENT practice with surgical facilities (n = 189) showed that there is no justification for adhering to literature reports of severe late complications with scarred tonsillar crypts and peritonsillar abscesses. No patient has developed any of these complications thus far. Following tonsillotomy, preoperative complaints (snoring, respiratory obstruction, dysphagia, and failure to grow) decreased considerably. Thus, tonsillotomy proved to be a suitable modality for treating tonsillar hyperplasia in early childhood. The prerequisite for long-term success is strict limitation of this intervention to the diagnosis of tonsillar hyperplasia. PMID- 12089814 TI - [Dilated pneumosinus of the frontal sinus]. AB - Pneumosinus dilatans is a rare disorder of the frontal sinus most commonly found in 20- to 40-year-old males. The deformity is caused by an abnormal enlargement of the frontal sinus by hyperpneumatization. A valve mechanism raising the pressure inside the sinus is regarded to be responsible for this condition. The diagnostic and surgical approach is presented in a 25-year-old male. A preoperative computed tomography (CT) scan in three planes illustrates the anatomical conditions and possible etiologic mechanisms. The surgical procedure is supposed to correct the esthetic deformity and the cause of the disorder. A bicoronal incision allows the best exposition of the frontal sinus. The osteoplastic correction is stabilized by titanium micro-osteosynthesis plates, which do not have to be removed. Functional endoscopic sinus surgery should be performed concomitantly in order to remove the cause of pneumosinus dilatans. PMID- 12089815 TI - [Ultrastructural changes of the nasal mucosa in primary ciliary dyskinesia]. AB - Primary ciliary dyskinesia syndrome (PCD) is a rare, autosomal receive disorder. Kartagener's syndrome is a subgroup of the PCD with situs inversus, bronchiectasis, and sinusitis. The symptoms results from an abnormal ultrastructural morphology of the cilia such as absence of dynein arms and other changes. As a consequence ciliary motility is disturbed. A 25-year-old man was examined because he suffered from recurrent severe pneumonia and Aspergillus infections of the lungs. On electron micrographs, ciliary abnormalities including deficiency of inner and outer dynein arms, dysmorphic outer dynein arms, and disorientation of the cilia were demonstrated. The diagnosis of PCD requires electron-microscopic investigations of the ciliated mucosa. Special attention should be given to ultrastructural changes of nasal or bronchial mucosa if a young patient suffers from recurrent severe respiratory infections. PMID- 12089816 TI - [Tumor of the nasopharyngeal wall. Hodgkin lymphoma (Hodgkin disease), nodular sclerosis, grade 1, stage Ia, primary manifestation of the nasopharyngeal wall]. PMID- 12089818 TI - [Lymphadenitis calls for strict follow-up]. PMID- 12089817 TI - [Unilateral ptosis--orbital complication? Aneurysm of the left internal carotid artery with complete oculomotor paralysis left]. PMID- 12089819 TI - [Bilateral bone anchored hearing aid management of children with external ear dysplasia and/or ear canal atresia]. AB - BACKGROUND AND OBJECTIVE: The literature gives hardly any information about the benefits for children of using a bilateral bone-anchored hearing aid (BAHA) as opposed to a bilateral conventional bone conducting hearing aid. PATIENTS AND METHODS: Within a period of 12 years, three children were selected for treatment with bilateral BAHA. At the time of surgery, the children were aged between 8 and 10 years. Two of the children suffered from Franceschetti syndrome and one child from Goldenhar's syndrome. The clinical course of these three patients is presented. RESULTS: In all cases, treatment with BAHA was beneficial according to subjective and objective criteria despite localized inflammatory complications in two of the three cases. CONCLUSIONS: The bilateral use of BAHA should be considered more frequently as an alternative to conventional bone conducting hearing aids even for children. PMID- 12089820 TI - [Characteristics of hearing aid management in infancy, early childhood and preschool age]. PMID- 12089821 TI - [Diagnostic value of color Doppler ultrasound diagnosis]. PMID- 12089822 TI - [The internet. Chances, risks and prospects for the surgical patient]. AB - The World Wide Web (WWW) serves consumers with an abundance of health information and services. This factor sets new accents in the health care system. The Internet can help to create an informed patient, who is actively involved in clinical decision-making processes ("shared decision making"). This has effects on the physician patient relationship. An informed patient demands better performance. The increased transparency and comparability of provider performance in patient care leads to an enhanced competition between providers. On the other hand, the Internet also bears certain risks: The quality of health information on the Internet is frequently described as unsatisfactory and patients rarely pay attention to quality markers of the information provided. It is necessary to encourage an active quality management for health information and services on the web. The Internet is not only a passive information resource but, above, all, an interaction medium. It can be harnessed to facilitate quality management. Intelligent technologies will gain increasing significance in the future, when the WWW develops towards a "semantic web". In particular, consumers will use intelligent computer programs, so-called "agents", to gather information or to evaluate the trustworthiness of health information providers. PMID- 12089823 TI - [Clinic communication and disease-oriented centers]. AB - German hospitals and surgical clinics/departments are facing far-reaching changes. One triggering factor is the imminent reorganization of hospital financing to a system of compensation, which is universally based on diagnosis related groups (DRGs) and entails a market-economy orientation in the hospital sector. Digital technologies, which facilitate making the necessary adjustments to clinic structures to meet forthcoming challenges, represent another element. The "digital transformation" of the hospital of the future takes place on three levels. The restructuring of the surgical realm runs rather a traditional course by increasing use of information technology, mostly to optimize documentation and existing procedures or to reduce costs. The second sphere reaches substantially further, encompassing reorganization of disease-oriented cooperation between the different medical specialties and enabling the establishment of suitably structured disease-oriented medical centers. This is followed by the third phase, which involves networking clinics or medical centers with private practitioners, aftercare and rehabilitation services, and other disease-oriented care providers. PMID- 12089824 TI - [Image fusion, virtual reality, robotics and navigation. Effects on surgical practice]. AB - In the new minimally invasive surgical era, virtual reality, robotics, and image merging have become topics on their own, offering the potential to revolutionize current surgical treatment and assessment. Improved patient care in the digital age seems to be the primary impetus for continued efforts in the field of telesurgery. The progress in endoscopic surgery with regard to telesurgery is manifested by digitization of the pre-, intra-, and postoperative interaction with the patients' surgical disease via computer system integration: so-called Computer Assisted Surgery (CAS). The preoperative assessment can be improved by 3D organ reconstruction, as in virtual colonoscopy or cholangiography, and by planning and practicing surgery using virtual or simulated organs. When integrating all of the data recorded during this preoperative stage, an enhanced reality can be made possible to improve intra-operative patient interactions. CAS allows for increased three-dimensional accuracy, improved precision and the reproducibility of procedures. The ability to store the actions of the surgeon as digitized information also allows for universal, rapid distribution: i.e., the surgeon's activity can be transmitted to the other side of the operating room or to a remote site via high-speed communications links, as was recently demonstrated by our own team during the Lindbergh operation. Furthermore, the surgeon will be able to share his expertise and skill through teleconsultation and telemanipulation, bringing the patient closer to the expert surgical team through electronic means and opening the way to advanced and continuous surgical learning. Finally, for postoperative interaction, virtual reality and simulation can provide us with 4 dimensional images, time being the fourth dimension. This should allow physicians to have a better idea of the disease process in evolution, and treatment modifications based on this view can be anticipated. We are presently determining the accuracy and efficacy of 4 dimensional imaging compared to conventional evaluations. PMID- 12089826 TI - [Hepatitis B. The HBV-infected surgeon--risk of infection for the patient]. PMID- 12089825 TI - [Intersections of reconstructive surgery in the area of regenerative medicine]. AB - Based on significant progress in biotechnology, "Regenerative Medicine" is evolving as a new discipline that could significantly change current therapeutic approaches. Medicine, stem cell biology, tissue engineering, nanotechnology, genomic research, material sciences, as well as system- and process-engineering come together to answer fundamental problems of human disease, including trauma and aging. The long-term goal is to create a profound knowledge of individualized therapy originating from nonbiological organ replacement, through hybridized systems, to fully biological implants and finally to human regenerative systems in vivo. Two model tissues, including heart valves and liver tissue are discussed in the context of in vivo and in vitro regeneration technologies. Extracellular matrix regulates tissue differentiation in at least three ways, including the biochemical composition of the matrix constituents, the 3-D organization or architecture, and via mechanical forces mediated to the cells by the matrix. The in vivo extracellular matrix constitutes a biopolymer which potentially plays a permissive role for tissue differentiation. PMID- 12089827 TI - ["Simplified Acute Physiology Score" (SAPS II) ina the assessment of severity of illness in surgical intensive care patients]. AB - INTRODUCTION: In 1993, Le Gall proposed a new Simplified Acute Physiology Score (SAPS II) to assess the severity of illness in intensive care patients and to predict the risk of hospital mortality using a large data base of more than 13,000 patients of different intensive care units. Up to the present time, no satisfactory form of validation for surgical intensive care patients has been available. We investigated the prognostic quality of this score system for the assessment of the severity of the illness in surgical intensive care patients. METHODS: Between October 1997 and December 1998, 310 consecutive patients were included in our study. We calculated the SAPS II score according to the published guidelines. Additionally, we calculated the risk of mortality by referring to the score. We used a receiver operating characteristics (ROC) analysis for statistical evaluation. RESULTS: The median duration of stay on our intensive care unit was 8.4 days. In all patients, the mean SAPS II was 29.9 +/- 12.7. In the survivors, calculation of mean SAPS II was 27.7 +/- 11.4. In the non survivors, mean SAPS II was 45.7 +/- 11.2. The predicted risk of mortality was 15.7%, the actual risk of mortality was 15.8%. CONCLUSION: The SAPS II score has proved to be a good prognostic instrument in surgical intensive care patients. SAPS II is particularly effective in providing an exact estimation of the risk of death, classifying patient groups in clinical studies, epidemiological investigations, and quality assessment. PMID- 12089828 TI - [How do general surgery patients experience the intensive care unit? Results of a prospective observational study]. AB - INTRODUCTION: Subjective experiences of patients during their stay in the intensive care unit (ICU) have so far rarely been described. The aim of this study was to analyze the experiences of patients during their stay in the ICU. METHODS: In a prospective study, 100 general surgical ICU patients were recorded consecutively. A questionnaire that covered a broad range of possible ICU experiences was handed out to patients shortly following their stay in ICU. At the same time, a questionnaire was given to the personnel of the ICU to investigate how well nurses and doctors were able to adopt the patients' perspectives of the ICU experience. RESULTS: Concerning the physical symptoms, insomnia was to the fore (67% of patients). Despite pain medication, 25% of patients reported severe pain. The main psychological symptom was a feeling of helplessness (29% of patients). As a general cause for concern, 48% of patients complained about limited mobility. The patients were critical of the presence of severely ill patients. The standards of nursing and medical attention, however, were judged very positively. The evaluation of the staff differed from the patients' experiences in many respects; the clearest differences concerned the items of pain, sleeping disorders and the observance of privacy. CONCLUSIONS: The study results led to several practical consequences in the quality of management procedure (e.g., the introduction of a thorough night's rest at the ICU, optimized information for patients). Additionally, we initiated further studies concerning the quality of life of ICU patients. PMID- 12089829 TI - [Antireflux surgery in Germany. Results of a representative survey with analysis of 2,540 antireflux operations]. AB - INTRODUCTION: Aim of this study was the evaluation of antireflux surgery in Germany. METHODS: An anonymous questionnaire including 288 structured items about diagnostic and therapeutic approaches, complications and mortality was sent to 33% randomly selected German general surgeons (n = 546) at the end of 2000. RESULTS: A total of 2,540 antireflux procedures was reported, 81% were performed laparoscopically; 65% were total, 31% partial fundoplications. The number of surgeons offering laparoscopic antireflux surgery increased from 0.3% in 1990 to 5% in 1995 and to 32% in 2000. Numerous modifications regarding esophageal mobilisation, crural repair, kind and extension of the wrap, use and size of a bougie, as well as an additional gastropexy were observed. Morbidity rates were significantly higher for open than for laparoscopic procedures (15.0% vs. 7.7%), mainly caused by wound healing problems (4.4% vs. 0.8%) and splenic lesions (3.1 vs. 0.6%). Gastric and esophageal perforations were similar in both groups (1.1 vs. 0.9%). Hospital mortality rate was 0.1%. We observed a frequency dependent learning curve regarding complication rates after laparoscopic antireflux surgery (< 11 fundoplication p.a.: 14.0%; 11-30 fundoplications: 7.3% (p = 0.05); > 30 fundoplications: 4.2% (p = 0.05%). Long-term-dysphagia occurred more often after 360 degrees-versus partial fundoplications (6.6% vs. 2.4%; p < 0.001) and after Nissen/Nissen-Rossetti--than after Floppy-Nissen-procedures (6.6% vs. 3.6%, p = 0.1). The recurrency rate was 9.3% without significant differences between the procedures. CONCLUSION: Laparoscopy has replaced the open technique. Different technical approaches significantly affect the outcome after laparoscopic antireflux surgery in Germany. PMID- 12089830 TI - [What is the value of the internet as a source of information for patients with inguinal hernias? First results of an observational study]. AB - INTRODUCTION: The increasing use of the internet has led to a variety of medical web pages and an increasing amount of information about hospitals. Little is known about the extent to which this new medium is already used by patients as a source of information. For patients with inguinal hernia, for example, a frequent surgical symptom with large method variety, the value of the internet as a source of information should be examined. METHODS: One hundred patients facing an elective inguinal hernia repair at the Surgical Department Charite Berlin took part in a prospective observation study (a questionnaire with 10 questions) between July 1999 and March 2001. The questions referred to the possibility of PC/internet access, other sources of information (e.g., general practitioner, friends, internet) as well as the criteria by which the clinic was chosen. Patients were asked to answer basic questions about the development and treatment of inguinal hernia and to give a self-assessment of their knowledge of inguinal hernia (0 = no knowledge; 10 = maximum knowledge). RESULTS: The questionnaire revealed that 39% of the patients (average age 47 +/- 16 years) had a PC in their homes, 24% of those with internet access; 11% had internet access at other places. It also showed that 53% of the patients sought medical information from their general practitioner, 29% from friends, and 18% from the internet. Young patients (median age 29 years), employees and patients with private internet access use the internet with significantly more frequency. Their medical knowledge as well as their self-assessment of medical knowledge was significantly higher (median 19 versus 7 points and median 8.6 versus 5.7, respectively) and they chose the clinic because of the information from their research on the internet. CONCLUSION: At present, the internet still plays a subordinate role as a source of information for patients with an inguinal hernia. The increasing presence of this medium as well as the higher internet acceptance of future patients indicates that it has a promising future as a source of medical information. PMID- 12089831 TI - [Pancreatic leakage after pancreas resection. An analysis of 345 operated patients]. AB - INTRODUCTION: Complications after pancreatic resections remain frequent despite a decreasing mortality. Pancreatic leakages represent a relevant part of those complications but data on risk factors for their occurrence are rare. We analyzed our experience with incidence, clinical course, and risk factors of pancreatic leakage in a large patient group. METHODS: We analyzed the prospectively documented perioperative data of 345 patients with pancreatic resections carried out between 1994 and 2001. Main indications for surgery were chronic pancreatitis (57%) and malignant tumors (37%). The following operations were performed: Whipple's operation 15%, pylorus-preserving pancreaticoduodenectomy 53%, duodenum preserving pancreatic head resection 19%, and distal pancreatic resection 13%. Risk factors were analyzed using uni- and multivariate methods. RESULTS: Postoperative mortality and complication rate were 2.9% and 41%, respectively. A pancreatic leakage occurred in 9.9%. In the majority of patients, pancreatic leakage was asymptomatic and controlled by prolonged drainage. However, one fourth of the patients with pancreatic leakage required reoperation. The mortality of pancreatic leakage was 12%. No patient with chronic pancreatitis died as a consequence of pancreatic leakage. Impaired preoperative renal function was the only risk factor for the occurrence of postoperative pancreatic leakage. CONCLUSIONS: Although easily managed in the majority of cases, pancreatic leakage still represents a relevant postoperative complication after pancreatic resection, especially in patients with malignant disease. Because of an increased risk of developing pancreatic leakage, an impaired renal function should be considered specifically in the perioperative management of the patients. PMID- 12089832 TI - [Factors influencing the development of incisional hernia. A retrospective study of 2,983 laparotomy patients over a period of 10 years]. AB - INTRODUCTION: Incisional hernia formation is one of the most frequent complications in visceral surgery requiring reoperation. Risk factors for incisional hernia formation and preventive strategies are not clearly defined. METHODS: In a retrospective study including 2983 patients over a 10-year period, the influence of demographic data, pre-, intra- and postoperative risk factors for incisional hernia development were evaluated. From the subgroups medical history, medication, laboratory values, indication, surgical technique, course of operation, postoperative course and wound healing, altogether 43 parameters were analysed. Statistical evaluation was performed using the chi 2-test according to Pearson, and binary logistic regression analysis. RESULTS: The mean incisional hernia incidence in the study was 4.3%. In the mean follow-up period of 21.1 months, the incisional hernia incidence was calculated at 9.8% using the Kaplan Meier estimate; for a 10-year period it reached 18.7%. The study revealed that 31.5% of all incisional hernias developed in the first 6 months after the operation, 54.4% after 12 months, 74.8% after 2 years and 88.9% after 5 years. Significant demographic factors influencing incisional hernia incidence were age (> 45 years) and male gender. The preoperative factors anaemia (Hb < 100 g/l) and BMI > 25, the intraoperative factors recurrent incision and previous laparotomy, and the postoperative factors catecholamin-therapy and disturbed wound healing were of significant influence. CONCLUSION: The calculated incisional hernia incidence for a 10 year period of almost 20% and the manifestation of 50% of all hernias more than 12 months after the operation, underline the necessity to intensify surgical research in the field of laparotomy healing. In comparison to demographic and endogenous risk factors, the surgical technique has less influence on laparotomy healing. Measures to ameliorate tissue perfusin seem to exert a positive influence on incisional hernia incidence. PMID- 12089833 TI - [Surgical reconstruction of high grade carotid stenosis: a safe procedure?]. AB - Carotid surgery is still controversial. Some large randomized trials have demonstrated the benefit of surgery in correlation to conservative treatment alone, but these positive results depend on how specific the diagnosis is and a low complication rate. This study presents the results of 2162 patients (male n = 1596 (74%), female n = 566 (26%), mean age 65 +/- 9 years), who underwent carotid surgery between 1990 and 1999. Forth-three percent of these patients had no ipsilateral neurological symptoms with high-grade carotid artery stenosis (Stage I). Thirty-eight percent appeared with prior ipsilateral TIA or PRIND- symptomatology (Stage II) and 19% suffered from stroke with persisting deficits (Stage IV). The operative technique of choice was thromboendarterectomy of the carotid bifurcation with vein-patch closure in 1967 patients (91%). In 1324 patients segmental resection of the internal carotid artery was performed. Carotid endarterectomies and other reconstructions for coronary artery disease including abdominal aortic aneurysm were combined during the same operation in 11% of the patients. The rate of postoperative ipsilateral neurological events was 4.1%. On the ontralateral side neurological symptoms appeared among 0.8%, and 0.4% of the patients had bilateral symptoms. Twenty patients (0.9%) died as a result of postoperative stroke. In relation to preoperative staging of the cerebrovascular occlusive disease in stage I, postoperative neurological symptoms appeared in 2.8% (mortality 0.6%), stage II in 5.7% (mortality 1.0%) and stage IV in 7.8% (mortality 1.2%) of the patients. These results confirm the importance of carotid reconstruction as a measure in the prevention of cerebral infarction in patients with asymptomatic or symptomatic high-grade carotid artery stenosis. The complication rate was lower than the data reported in the literature and the results were clearly better than under conservative treatment alone. In our opinion, the indication for carotid artery reconstruction should be made by a team of vascular surgeons, neurologists and neuroradiologists taking all patient specific factors into consideration. Only by optimal patient selection and minimal complication rates will a significant benefit for the patient be achieved. PMID- 12089834 TI - [Computer-assisted case-based education in surgery]. AB - INTRODUCTION: Computer technology is well established in the fields of medical diagnosis and monitoring. In recent years, there has also been an increase in the use of computers for the purpose of medical education. METHODS: With the aid of the "author's-system" CASUS, investigation of a case of acute abdominal pain was integrated into the practical medical training for students in the 4th. year of clinical studies. For patient history, clinical examination and laboratory tests, the method involves the use of 41 cards with pictorial and acoustic information (text, graphics, sound recordings and videos), including 23 questions to be answered by the students. At the end of the session, the acceptance, effectiveness and practicability of the computer-based training was evaluated by a questionnaire. RESULTS: In 356 students, a high rate of acceptance and satisfaction was achieved (positive judgement of various aspects: 55%-98%). Technical difficulties which occurred initially were overcome during the course of the evaluation. Some questions were answered more positively by female students. CONCLUSION: Most students share the view that computer-based training, though not able to take the place of bedside teaching, is a valuable addition to the armamentum of medical teaching. PMID- 12089835 TI - [Documentation of surgical performance--does more really help more? Comparison of the effects of maximum and limited documentation depth of clinical patient data on theoretical revenue volume of a surgical clinic after introduction of the DRG based reimbursement system]. AB - INTRODUCTION: The forthcoming introduction of a DRG-based account system in Germany aims at higher transparency and economic efficiency, particularly in the sector of in-patient health care. The availability of documentation of the highest quality, taking into account all potentially relevant diagnoses, appears to be the best method for achieving maximum revenue in individual surgical units. The aim of the study was to determine the relevance of various degrees of documentation depth on calculated DRG-based revenue. Furthermore, we evaluated whether improvements in the quality of documentation can be realized in current hospital organization. METHODS: In a prospective study, clinical data from 402 in patients were collected and revenues were calculated based on the Australian Refined DRG system. Various qualities of documentation were defined. In order to find the medical sectors most sensitive to "under-documentation", homogenous cases were classified into 23 treating groups, according to diagnosis. RESULTS: In 267 cases, maximum revenue was determined only by one main diagnosis, while better results could be achieved in 137 cases (34%) by extended documentation quality. Half of this gain could only be achieved by an independent medical documentation specialist. An upper limit of documentation intensity (number of diagnoses) could be defined. Maximum gain did not require maximum number of diagnoses. CONCLUSIONS: Documentation depth has an important influence on the calculated revenue of surgical therapy based on AR-DRG system. The quality and depth of the documentation is not, in itself, sufficient. In order to be really effective, it requires the highest degree of professionalism from hospital staff. PMID- 12089836 TI - [3-dimensional computer animation--a new medium for supporting patient education before surgery. Acceptance and assessment of patients based on a prospective randomized study--picture versus text]. AB - INTRODUCTION: The rigorous implementation of clear preoperative information is mandatory for the patient's understanding, acceptance and written informed consent to all diagnostic and surgical procedures. In the present study, I evaluated whether new media are suitable for conveying basic information to patients; I analysed the merits of computerized animation to illustrate a difficult treatment process, i.e., the progressive steps of a thyroid operation, in comparison to the use of conventional flyers. METHODS: 3D animation software was employed to illustrate the basic anatomy of the thyroid and the larnyx; the principle of thyroidectomy was explained by visualizing the surgical procedure step by step. Finally, the possible complications that may result from the intraoperative manipulations were also visually explained. Eighty patients entered a prospective randomisation: on the day before surgery, group 1 watched the computer animation, whereas group 2 was given the identical information in a written text (= standard flyer). The evaluation included a questionnaire with scores of 1-5, rating the patients' understanding, subjective and objective knowledge, emotional factors like anxiety and trust, and the willingness to undergo an operation. RESULT: Understanding of and subjective knowledge about the surgical procedure and possible complications, the degree of trust in professional treatment, the reduction in anxiety and readiness for the operation were significantly better after watching the computer animation than after reading the text. However, active knowledge did not improve significantly. The interest in the preoperative information was high in both groups. The benefit of computer animation was enhanced in a second inquiry; patients who had only read the text had a significant improvement in parameters after an additional exposure to the video animation. CONCLUSION: Preoperative surgical information can be optimized by presenting the operative procedure via computer animation. Nowadays, several types of new media such as the world wide web, CD, DVD, and digital TV are readily available and--as shown here--suitable for effective visual explanation. Most patients are familiar with acquiring new information by one of these means. An appropriately designed 3D repre-sentation is met with a high level of acceptance, as the present study clearly shows. Modern patient-based information systems are necessary. They can no longer be the sole responsibility of the medical profession, but must be on the agenda of hospital managements and of medical care systems as well. PMID- 12089838 TI - [Keloid of the earlobe after ear piercing. Not only a surgical problem]. AB - Earlobe keloids are benign, fibrous proliferations that show a high rate of recurrence of up to 80% following surgical excision. Traumas to the earlobe such as ear piercing, burns or surgical interventions are important in the pathogenesis of the disease. In addition to surgical keloid excision and reconstruction of the earlobe, several adjuvant therapeutic concepts have been described to prevent recurrence. Here we present the case of a female patient who suffered from severe bilateral keloid development after piercing of both ears. The report gives an overview of the relevant therapeutic concepts in the treatment of earlobe keloids and their possible complications. In addition, the question of written informed consent before ear piercing is discussed. PMID- 12089837 TI - ["Multimedia symposium wares". An enrichment of medical and graduate education?]. AB - The continual development of the internet has supported the spread of surgical knowledge by electronic means. High quality products have to be offered from a software as well as a contents point of view. The question as to whether these new media and their contents have a real value for efficient and motivating use in medical education needed to be answered by first assessing a quality profile for the development of surgical educational modules which were then evaluated on the basis of so-called "symposia ware". First, the reactions and opinions of physicians at 47 universities were assessed by a standardized questionnaire concerning their demands on multimedia teaching/learning modules. Several different aspects of technique, content, presentation, didactics and background knowledge were analyzed. In a second step, their opinions were evaluated concerning two applications (symposia implemented on CD-ROM as a slideshow with original slides and audio) with surgical and gastroenterological contents by standardized questionnaire. Questions concerning personal background such as educational status and experience with computers, e-mail and the internet on one hand and the CD-ROM itself concerning content, relevance for daily clinical work and continuing medical education as well as the quality of the application on the other hand, were evaluated using marks (1 = best, 6 = worst). A total of 320 physicians participated in the first part of the interview. Of these, 93% were equipped with computers in hospital as well as privately. The Internet was used by 90% of them. The majority declined a full text presentation as well as the application of scroll fields. The participants rather favored the integration of text, pictures, animations and videos. Furthermore, 95% demanded the provision on the internet. Thirty-seven colleagues in their 5th (1-11) year of training were interviewed, and of those, 27 were working in a surgical department and 10 in a medical department. Individual computer knowledge was rated with a median of 3. This revealed that 60% were equipped with computers in hospital as well as at home, the remaining 40% had computers either in hospital or at home. All participants used the internet. In total, 57% had experience with "Symposia ware". The rating of the "Symposia ware" itself was positive. Relevance and applicability of a slideshow for imparting knowledge were rated with a median of 2. This showed that 81% would buy the CD-ROM in principle, and 89% would spent up to 50 EUR. Quality, language, content and user-friendliness were all rated 2. Physicians frequently use computers and the internet. All this indicates a high degree of acceptance of electronic teaching/learning modules in medical education. A uniform structure of contents as well as a platform-independent, web based presentation is appreciated. To enhance illustration, a picture and video oriented visualization should be chosen. Overall, "symposia ware" is rated positively. It should cost no more than 50 EUR and it represents a valuable source of information for physicians. PMID- 12089840 TI - [In the borderline of medicine]. PMID- 12089839 TI - [Donor organ harvesting]. PMID- 12089841 TI - [Functional gastrointestinal diseases]. PMID- 12089842 TI - [A child is born--we can afford a celebration!]. PMID- 12089843 TI - [Treatment of acute pancreatitis]. AB - BACKGROUND: The role of surgery in the treatment of acute pancreatitis is still unclear. The aim of this study was to evaluate and compare our results with different treatment strategies described in the literature. MATERIAL AND METHODS: From 1980 to 1998, 376 patients, 195 women and 181 men, median age of 62.5 years (15-96 years) with acute pancreatitis were treated in our hospital. The classification of the pancreatitis was based on Ranson signs (more than four defined as serious). Patients with septic necrosis/abscesses were treated with percutaneous drainage and irrigation. RESULTS: The disease was caused by gallstone in 58%, ethyl in 13%, unknown cause in 14%, and miscellaneous in 15%. In 50 patients (13%) the pancreatitis was serious. Ten patients (2.6%) died in hospital. Among all patients with serious/haemorrhagic pancreatitis the mortality was 18%. INTERPRETATION: Intensive supportive care and broad spectrum antibiotics combined with percutaneous drainage and irrigation of infected intraabdominal content in haemorrhagic pancreatitis seem to give the same results as open surgery. PMID- 12089844 TI - [Antineutrophil cytoplasmic autoantibodies in systemic vasculitis]. AB - BACKGROUND: Antineutrophil cytoplasmic autoantibodies (ANCA) are directed against antigens located in the cytoplasm of neutrophil granulocytes and monocytes. Detection of ANCA has proved to be a useful diagnostic tool for a group of systemic vasculitis, especially Wegener's granulomatosis. Both indirect immunofluorescence (IIF) and ELISA have been used to detect ANCA. MATERIAL AND METHODS: In this study, samples from 319 patients tested by both immunofluorescence and ELISA were evaluated; 27 of these were diagnosed with Wegener's granulomatosis. RESULTS: The diagnostic sensitivity for Wegener's granulomatosis was 70% for C-ANCA and 63% for PR3-ANCA. The specificity was 97% and 99% respectively. Positive predictive value for the diagnosis of Wegener's granulomatosis in our population was 68% for C-ANCA and 90% for PR3-ANCA. Negative predictive value was 97% and 97% respectively. INTERPRETATION: We recommend that immunofluorescence is used for screening when an ANCA-associated vasculitis is suspected. However, a number of antigen specificities can provide the immunofluorescence patterns, and for this reason we recommend PR3-ELISA and MPO-ELISA tests whenever the immunofluorescence test is positive. PMID- 12089845 TI - [Spasticity treated with selective posterior rhizotomy]. AB - BACKGROUND: Spasticity is often seen in patients with central nervous lesions. Some patients with severe spasticity are not optimally treated with physiotherapy and medication. MATERIAL AND METHODS: We present a case history of a 41-year-old woman with multiple sclerosis and severe painful spasticity in her lower limbs. Her spasticity did not respond to treatment with physiotherapy, spasmolytic medication, botulinum toxin A, intrathecal baclofen or epidural spinal cord stimulation. RESULT: The patient was treated with selective posterior rhizotomy S1-L1. Section of 60% of the rootlets on the right side and 40% on left the side resulted in a good outcome with less spasticity and pain. Finally her contractures were treated with tenotomy and myotomy, also with good functional result. INTERPRETATION: Patients suffering from severe painful spasticity and who do not respond to physiotherapy in combination with other spasmolytic medication should be considered for surgical treatment. In some patients posterior rhizotomy is the treatment of choice. PMID- 12089846 TI - [Diagnosis and treatment of penile cancer]. AB - BACKGROUND: Penile cancer is a rare malignancy in Norway with about 40 new cases each year. MATERIAL AND METHODS: An overview on diagnosis and treatment of penile cancer is given and the guidelines from the European Association of Urology are presented. RESULTS: Phimosis and poor genital hygiene are pre-disposing conditions for penile cancer. Condylomata acuminatum and leukoplakia should be regarded as premalignant lesions. The superficial form of penile cancer should be treated by laser, surgery or radiotherapy. When the cancer infiltrates into corpus cavernosum or corpus spongiosum, or the tumour displays a poorly differentiated histology, a partial or total amputation of the penis has to be performed. Inguinal lymphadenectomy is recommended in patients presenting with a tumour > or = pT2 or if the histology reveals a moderately or poorly differentiated cancer. Five-year survival rate is about 80% for patients with localised tumour, and about 50% in patients with regional lymph node metastasis. INTERPRETATION: We recommend that the treatment of penile cancer is performed in the regional hospitals. PMID- 12089847 TI - [Regular prescription of narcotics and sedatives to drug-addicted patients in general practice]. AB - BACKGROUND: A nation-wide rehabilitation programme with legal prescription of methadone and buprenorfine has been organised for drug-addicted patients, but the number of patients waiting for such treatment is increasing. The Norwegian health authorities have discouraged general practitioners from prescribing quotas of drugs to drug-addicted patients waiting for admission to these programmes. MATERIAL AND METHODS: 15 patients, 31 to 42 years old, with 15 to 25 years of drug abuse are presented. All patients are waiting for admission to a rehabilitation programme or institution. They have been treated individually with specified quantities of medication in a private general practice in Oslo, Norway. RESULTS: After 1 to 3 years of treatment, all patients are alive and live by themselves in regular apartments. Their social condition have improved and they have better contact with their families. There were no conflicts with public authorities, and abuse of illegal drugs was reduced. INTERPRETATION: The results are in accordance with those obtained in centres for rehabilitation of drug addicted patients. General practitioners should be encouraged to provide intermediate treatment of drug-addicted patients waiting for admission into rehabilitation programmes. PMID- 12089848 TI - [Rooming-in in the maternity ward--are mothers satisfied?]. AB - BACKGROUND: 24-hour rooming-in in Norwegian maternity wards has been introduced as one of the "10 steps to successful breastfeeding", the basis for WHO/UNICEF Baby-Friendly Hospital Initiative. This step, optimal in order to get lactation off to a flying start, has lead to some worry about maternal fatigue. MATERIALS AND METHODS: Self-administrated questionnaires to health care personnel as well as to mothers in all Norwegian maternity wards were used to evaluate the impact of the Baby-Friendly Hospital Initiative. The same mothers were interviewed again 15 months later. RESULTS: Many (52%) felt tired after giving birth, but most (85%) were content with the care received. There was no difference in tiredness between mothers who chose to keep their babies with them and those who did not. Only 3.3% cited this at a main reason for tiredness. INTERPRETATION: Most mothers felt positive about keeping their infants with them at night. This demonstrates that Norwegian maternity wards generally have been able to implement 24 hour rooming-in in a caring way. Staff should offer to look after fussy babies without mothers demanding it and give the new mothers care and support in all possible ways. PMID- 12089849 TI - [Are results of large clinical drug trials valid for both men and women?]. PMID- 12089851 TI - [Functional dyspepsia--a psychosomatic disease]. AB - About 4% of consultations in general practice involve patients with upper gastrointestinal complaints. Evidence of peptic ulcer disease is identified in only 20-30% of the patients. No organic explanation of the symptoms is found using endoscopy in 20-50%; these patients are given the diagnosis non-ulcer dyspepsia or functional dyspepsia. Hypersecretion of gastric acid and the bacteria Helicobacter pylori, which seem to be major aetiologic factors in duodenal ulcer, are not important in functional dyspepsia. In these patients gastric motor abnormalities and visceral hypersensitivity are the most important pathophysiological mechanisms. The gastric motility is influenced by stress; there is a strong relationship between anxiety, depression and functional dyspepsia. Antacids, H2-blockers and prokinetics are hardly more effective than placebo in patients with functional dyspepsia, while antidepressants have been proven effective in reducing dyspeptic symptoms. Likewise, psychological treatment like bio-feedback, stress management, interpersonal psychotherapy and cognitive therapy has also been proven effective in reducing dyspeptic symptoms in patients with functional dyspepsia. PMID- 12089850 TI - [Irritable bowel syndrome--a multifactorial disease in children and adults]. AB - Irritable bowel syndrome is the most frequent gastrointestinal disorder in Norway. Though there has been huge research activity in the field, no proven single aetiology or effective treatment has emerged. Consensus-based clinical diagnostic criteria have not yet brought diagnostic clarity. Possibly, there are dysfunctions in the processing of sensory stimuli in the "brain-gut" axis that may cause visceral hypersensitivity and secondary motility changes. In some patients, a multifactorial explanation of the mechanisms is useful, including stressful life events or other psychological factors. Psychiatric co-morbidity is probably the most important maintaining factor. Irritable bowel syndrome may serve as a model for the study of the interaction between biological, psychological and social factors in functional disorders. A good therapeutic relationship between the physician and the patient is an important element in the treatment approach. PMID- 12089852 TI - [Chronic pelvic pain in women]. AB - BACKGROUND: This paper reviews empirical and clinical evidence of the aetiology and treatment of medically unexplained chronic pelvic pain in women. MATERIAL AND METHODS: Clinical experience from an ongoing randomised treatment trial supplemented by computer-assisted reviews of studies obtained by a Premedline and Medline search (1996 to February 2002) and data from the Cochrane Database of Systematic Reviews and the EBM database of Abstracts of Reviews of Effectiveness. RESULTS: The aetiology of medically unexplained chronic pelvic pain is disputed but likely to be multifactorial. A history of interpersonal difficulties and a stressful life is common, and comorbid psychiatric disorders occur frequently. No treatment of choice emerges from the few controlled treatment trials, though a flexible biopsychosocial approach seems the most promising. INTERPRETATION: Empathic medical evaluation and follow-up within a biopsychosocial framework is recommended. Analgesic, hormonal and, if appropriate, surgical treatment can relieve pain. Additional benefits may be obtained by adding sensory awareness directed physiotherapy aimed at changing painful muscle tensions, body attitude, movement pattern and dysfunctional respiration pattern. Co-morbid psychiatric disorders should be diagnosed and treated. Cognitive-behavioural stress management intervention aimed at improving coping with pain and current life situation may be indicated in a subsample of patients. PMID- 12089853 TI - [From "quota prescription" to drug-assisted rehabilitation--experiences with the temporary guidelines]. PMID- 12089854 TI - [Nutrition is medicine, too--and education is necessary]. PMID- 12089855 TI - [Research and connections]. PMID- 12089856 TI - [Application routines for support from the educational fund III are not satisfactory]. PMID- 12089857 TI - [Systematic evaluation of sickness certification II]. PMID- 12089858 TI - Multiple endocrine neoplasia syndromes. PMID- 12089859 TI - How to detect pheochromocytomas?--the diagnostic relevance of plasma free metanephrines. AB - Pheochromocytomas are chromaffin cell tumors, mostly originating from the adrenal medulla, and represent a rare cause of hypertensiondue to excessive production of catecholamines (norepinephrine and/or epinephrine). More than 10% occur in families with multiple endocrine neoplasia type II, von Hippel-Lindau disease, neurofibromatosis type I, and familial carotid body tumors. Since approximately half of the afflicted patients present without or with only episodic hypertension, detailed clinical evaluation and sensitive biochemical tests are mandatory for the diagnosis, which relies on the detection of increased catecholamine production. Commonly employed tests such as the measurement of free catecholamines in plasma and urine or of their metabolites, vanillylmandelic acid and total metanephrines (= free + conjugated normetanephrine and metanephrine) in urine, suffer from interference from external factors and sometimes low clinical sensitivity and/or specificity. Recent technical advances now allow us to measure plasma free (unconjugated) metanephrines, thus increasing clinical sensitivity and specificity to close to 100%. Plasma free metanephrines offer the following advantages for the detection of pheochromocytomas: (i) independence of short-term changes in catecholamine secretion which may result from change of posture, exercise or intraoperative stress, (ii) information on long-term increase of catecholamine production, (iii) tight correlation with tumor mass, and (iv) only minor interference from drugs. This method does not need time-consuming standardized procedures for blood sampling, which are a prerequisite for the determination of free catecholamines. In conclusion, it is therefore recommended to use plasma free metanephrines--after meticulous clinical screening--as the first-line biochemical test for detecting pheochromocytomas. PMID- 12089860 TI - Multiple endocrine neoplasia type 1 (MEN1) in Austria. AB - Multiple endocrine neoplasia type 1 (MEN1) is a rare cancer predisposition syndrome. It results from the autosomal dominant inheritance of inactivating germ line mutations of the MEN1 tumor suppressor gene. Mutation carriers are prone to develop tumors, preferentially, of the parathyroid and anterior pituitary glands as well as the enteropancreatic endocrine tissues. Because such tumors also occur without the MEN1 context, we have set up a molecular genetic screening program in Austria to discriminate between heritable and non-heritable tumor forms. Following the recognition of a MEN1-specific germ-line mutation in a tumor patient, we extend the screening to all first-degree relatives. To date, we have studied 42 individuals by sequencing the coding exons 2 to 10 of the MEN1 gene. A germ-line mutation was discovered in four of seven families suspected, clinically, to have MEN1, and in 3 of 22 (13.6%) patients with a presumed sporadic endocrine tumor. The respective mutations were also detected in three first-degree relatives of whom only one 6-year-old boy was asymptomatic at the time of investigation. The possibility to clearly discriminate between genetically predisposed and non-predisposed individuals has a significant impact on the diagnosis and clinical management of both patients and their relatives. Both symptomatic and asymptomatic mutation carriers can be closely monitored, thereby allowing early recognition and treatment of developing tumors. Non affected relatives, on the other hand, do not require further controls. Finally, this approach also provides the information necessary for reliable genetic counseling. PMID- 12089861 TI - Multiple endocrine neoplasia 1--current recommendations for diagnosis and treatment. AB - BACKGROUND: The principally affected glands in the MEN 1 syndrome (parathyroids, pancreas, pituitary and adrenal glands) are often diffusely or multi-centrically involved, making different therapeutic approaches necessary. METHODS: In a retrospective analysis of 10 patients with genetically proven (n = 7) or clinically suspected (n = 3) MEN 1 syndrome, recommendations for diagnosis, timing of interventions and surgical procedures are reviewed. RESULTS: All patients had primary hyperparathyroidism (PHPT). An extended bilateral exploration localized 4 or more enlarged glands in 6 patients and subtotal parathyroidectomy (SPTX) was performed. In 4 patients, only one (n = 2) or two (n = 2) enlarged glands were removed. Two patients were reoperated for persistent PHPT and one patient developed recurrent PHPT. In 3 out of 6 patients, neuroendocrine pancreatic tumors were the first manifestation. 2 patients had solitary, one patient multiple benign and one patient multiple malignant insulinomas. Tumors were removed by enucleation, distal pancreatic resection or a combination of both. Out of the 2 patients with gastrinomas, one underwent partial pancreatoduodenectomy and the other has refused operation up to now. During follow-up, no persistence or recurrence of hormone excess was diagnosed. Three patients had non-functioning bilateral lesions of the adrenal glands, and one of these additionally had a small, clinically insignificant pheochromocytoma. Adrenalectomy was performed during pancreatic surgery in 2 patients, and endoscopically in one patient. Pituitary tumors were treated in 3 patients. CONCLUSION: A high index of clinical suspicion, biochemical screening and menin gene testing, according to current guidelines, is mandatory for early diagnosis of MEN 1. In PHPT with multiglandular involvement and neuroendocrine pancreatic tumors, meticulous surgery can achieve a long-term cure in the majority of patients, with low morbidity. PMID- 12089863 TI - Recommendations for reporting C cell pathology of the thyroid. AB - BACKGROUND: Calcitonin screening programs have proved to be effective in early detection of medullary thyroid carcinoma, not only in patients with known risk factors for the development of hereditary tumors. Thus, more thyroidectomies, based on an abnormal pentagastrin test, can be expected. Here we give summarizing recommendations for reporting C cell pathology. METHODS: All patients underwent total thyroidectomy and were tested for germ-line mutations in the RET Protooncogene. The entire surgical specimens were blocked and C-cell disorders were assessed using conventional histology and immunohistochemistry. RESULTS: Among 110 patients with an abnormal pentagastrin test, 60 (55%) had medullary thyroid carcinoma (T1 34% [n = 37], T2 14% [n = 16], T4 6% [n = 7]), and 50 (45%) had C cell hyperplasia only. C cell hyperplasia accompanying medullary thyroid carcinoma was found in 13 of 15 familial and in 28 of 45 sporadic patients. All C cell changes were found in the upper two thirds of the thyroid lobes and 83% of the medullary thyroid carcinomas could be identified with frozen sections. CONCLUSION: 1. Abnormal pentagastrin stimulation is always associated with either medullary thyroid carcinoma or C cell hyperplasia. 2. Blocking of the entire upper two thirds of the thyroid lobes is essential for reliable detection of C cell hyperplasia and small medullary thyroid carcinomas. 3. Most medullary thyroid carcinomas can be detected with intraoperative frozen sections. 4. The presence of C cell hyperplasia should always be reported; however its usefulness for indicating familial risk is limited and its role as a preneoplastic condition in patients without RET-protooncogene mutations remains to be elucidated. PMID- 12089862 TI - Screening for medullary thyroid carcinoma: experience with different immunoassays for human calcitonin. AB - Calcitonin measurements in patients with nodular thyroid disease are helpful for early diagnosis and therapy of medullary thyroid carcinoma. We compared three commercial calcitonin assays routinely used: "CIS" (France), "Medgenix" (Belgium) and "Nichols-Advantage" (USA). In addition, we evaluated a two-step modification of the Medgenix-test and an enzyme immunoassay from Sangui (Japan). Method comparison studies revealed deviations from linear relationships between all routine assays. While histograms of CIS and Nichols results were similar at low concentrations with their highest frequency below 1 pg/ml, Medgenix showed a broad peak around 3 pg/ml. Correlation coefficients were 0.69 (CIS versus Medgenix) and 0.91 (CIS versus Nichols). In thyroidectomized patients, calcitonin was not detectable with CIS and Nichols, but the Medgenix test, which was more susceptible to interference, measured about 6 pg/ml. The immunoassay of Sangui showed insufficient analytical sensitivity. About 70-80% of calcitonin levels initially > 10 pg/ml were reproduced in the basal levels of the subsequent pentagastrin test. Average ratios (stimulated/basal level) were slightly higher for CIS and Nichols than for Medgenix. Prediction of a pathological stimulation from basal calcitonin was insufficient with CIS and Medgenix assays. If a calcitonin concentration of > 100 pg/ml (CIS) is considered as an indication for surgery, equivalent values are 71 pg/ml for Medgenix, and 96 pg/ml for Nichols. Using these criteria, about one third of patients who underwent pentagastrin stimulation showed pathological reactions. Reliable and sensitive calcitonin assays used for the screening of sporadic medullary thyroid carcinomas in patients pre-selected by nodular goiter are important for health and economic reasons, because they enable early therapy. PMID- 12089864 TI - Medullary thyroid carcinomas in cell culture--models for future therapies. AB - We report the successful establishment of seven human medullary thyroid carcinomas (MTC) as continuous cell lines. Characteristic features--such as the presence of neuroendocrine granules--and the positive immunoreactivity to antibodies to CT, CGRP, GRP, SRIF, 5-HT, NSE, PHE, LK2H10, ER and Pgr were followed throughout the cultivation. An overexpression of the antiapoptotic gene bcl-2 was detected in the cell lines. Deregulation of apoptosis plays an important role in multistep tumorigenesis. MTCs are known for the phenomenon of bcl-2-based chemo- and radioresistance. Our studies focus on influencing the growth rates and modulating the apoptotic rates by treatment with proliferation modifying substances and anticancer drugs. Our MTC cell lines are useful models for these in vitro studies. PMID- 12089865 TI - Eating disorders. What's new? PMID- 12089866 TI - Leptospiral carriage by mice and mongooses on the island of Barbados. AB - Leptospirosis is a zoonotic disease, maintained by chronic infection of the kidneys of reservoir animals, usually small mammals. Infection in humans is acquired from direct or indirect exposure to the urine of infected animals. Leptospirosis has a high incidence in tropical regions, and has been studied extensively in several Caribbean countries. We studied the carriage of Leptospira serovars by two small mammals which are potential maintenance hosts of the disease in Barbados. A total of 136 mongooses (Herpestes auropunctatus) and 97 mice (Mus musculus) were caught in live traps. Leptospiral antibodies were detected by microscopic agglutination test (MAT) using antigens representing 12 serogroups, and kidney tissues were inoculated into polysorbate medium for isolation of leptospires. The seroprevalence (at a titre of > or = 100) in mice was 28.2% (24/85, 95% CI 19.0, 39.1) and in mongooses 40.7% (48/118, 95% CI 31.7, 50.1). In mice, antibodies were detected predominantly against serogroups Ballum and Autumnalis, while in mongooses the predominant serogroup was Autumnalis. Leptospires were isolated from 28 mice (28.9%, 95% CI 20.1, 39.0) and from 4 mongooses (2.9%, 95% CI 0.8, 7.4). Mouse isolates were identified as serovars arborea (17) and bim (7). As in other parts of the world, common house mice (Mus musculus) represent a significant reservoir of leptospirosis. Although carriage of the Ballum serovar, arborea, was not unexpected, this represents the first time that an animal reservoir of serovar bim has been identified. This is significant because bim causes about 63% of human leptospirosis in Barbados, and control efforts and education for prevention can now be targeted at a specific reservoir. PMID- 12089867 TI - Evaluation of location of mandibular and mental foramina in dry, young, adult human male, dentulous mandibles. AB - The present study was undertaken to determine the precise location of the mental and mandibular foramina and the surrounding anatomic landmarks. Measurements were made on both right and left sides of each of 34 dried young Turkish adult (30-40 years old) human male mandibles. The mandibles were moderately robust and were from dentulous subjects. The results obtained in the study showed that the distance of the mandibular foramen to the angle of the anterior ramus were 16.9 mm on the right and 16.78 mm on the left. The distance to the posterior side of the ramus was 14.09 mm on the right, and 14.37 mm on the left. The narrowest anteroposterior diameters were 32.8 mm on the right and 32.05 mm on the left. The mandibular foramen is 0.5 mm posterior to the centre of the ramus on the right and 0.75 mm on the left. The distance of the lowest point of mandibular notch to the foramen was 22.37 mm on the right and 22.17 mm on the left. The distance from the mandibular foramen to the inferior border of the ramus in the mid position of the ramus was 30.97 mm on the right and 29.75 mm on the left. The average angle of the mandible (gonial angle) was 120.17 degrees on both sides. The distance of the mental foramen to the inferior border of the mandible was 14.61 mm and 14.29 mm on the right and left, respectively. Its distance to the superior border was 13.62 mm on the right and 14.62 mm on the left. The horizontal dimensions of the mental foramen was 2.93 mm on the right, 3.14 mm on the left and its vertical dimension was 2.38 mm on the right and 2.64 mm on the left. The mental foramen was located below the root of the second premolar in 21 bones (61.76%) on the right and 17 bones (50%) on the left while in the remaining mandibles, the foramen was present between the roots of the first and second premolars in 13 bones (38.2%) on the right and in 17 (50%) on the left. The average angle of the mental foramen was 96.82 degrees. PMID- 12089868 TI - Iatrogenic ulnar neuropathies post-pinning of displaced supracondylar humerus fractures in children. AB - A retrospective review of 141 displaced supracondylar fractures in children at the Bustamante Children's Hospital and the University Hospital of the West Indies from 1994 to 1999 revealed ten ulnar nerve palsies. Of the 141 supracondylar fractures, 27 were treated with open reduction and internal fixation, while 114 had closed reduction and percutaneous pinning. All fractures were fixed with crossed Kirschner wires. Of the ten cases, the ulnar nerve was explored in two cases; the medial pin was removed in two cases, while the other six cases were observed. Follow-up ranged from three to 18 months. Full nerve recovery occurred in all cases except one in which there was partial return of function. Recommendations are made regarding the management of these injuries. PMID- 12089869 TI - Nosocomial infections in an intensive care unit in a private hospital. AB - The nosocomial infection rate in the intensive care unit (ICU) of a private hospital was assessed during an 18-month survey. From 629 admissions to the ICU, 139 hospital-acquired infections were identified. The rate was 22.1% compared to the overall nosocomial infection rate of 15.3% for the entire hospital. In the ICU, the main infections occurred in the respiratory tract, 41 (29.5%), followed by surgical wounds, 35 (25.2%), urinary tract, 28 (20.1%) and the blood stream, 24 (17.3%). From 165 bacterial isolates, 80% of isolates were gram-negative rods, with P aeruginosa, 48 (36.6%), being the predominant gram-negative isolate followed by Klebsiella pneumoniae, 27 (20.6%), and Enterobacter sp, 22 (16.8%). The main gram-positive isolates were S aureus, 23 (41.8%), coagulase-negative Staphylococci, 17 (30.9%), and Enterococci, 11 (20.0%). Of the 23 S aureus strains, 15 (65.2%) were methicillin-resistant (MRSA), (8 MRSA were from surgical wounds, 5 from the respiratory tract and 2 from infected urine). Only 2 of the 17 (11.8%) coagulase-negative staphylococci were methicillin-resistant, and both were isolated from wounds. Resistance to ampicillin and augmentin (amoxicillin clavulanic acid) was high, 81.9% and 55.4%, respectively. Gentamicin, aztreonam, piperacillin and piperacillin-tazobactam showed resistance rates of less than 15%. Infection control measures aimed at reducing nosocomial infections at the hospital are often frustrated by apathy of hospital administrators who apparently are insensitive to the high nosocomial infection rate. Effort by the infection control team through seminars, lectures and newsletters have begun to show improvements in attitude and awareness of staff to infection control and preventative measures within the institution. PMID- 12089870 TI - Intestinal parasites among young children in the interior of Guyana. AB - Intestinal parasites contribute greatly to morbidity in developing countries. While there have been several studies of the problem in the Caribbean, including the implementation of control programmes, this has not been done for Guyana. The aim of this study was to determine the prevalence of intestinal parasites among young children in a town located in the interior of Guyana. Eighty-five children under the age of 12 years were studied prospectively for intestinal parasites in Mahdia, Guyana. Stool samples were transported in formalin to the Department of Microbiology, The University of the West Indies, Jamaica, for analysis using the formalin-ether concentration and Ziehl-Neelsen techniques. Data on age and gender of the children were recorded on field data sheets. At least one intestinal parasite was detected in 43.5% (37/85) of the children studied and multiple parasitic infections were recorded in 21.2% (18/85). The most common intestinal helminth parasite was hookworm (28.2%; 24/85), followed by Ascaris lumbricoides (18.8%; 16/85) and then Trichuris trichuria (14.1%; 12/85). Among the protozoan infections Giardia lamblia was detected in 10.5% (9/85) of the study population while Entamoeba histolytica appeared rarely. All stool samples were negative for Cryptosporidium and other intestinal Coccidia. There was no predilection for gender with any of the parasites. The pattern of distribution of worms in this area of Guyana was unlike that seen in other studies. Hookworm infection was the most common among the children and a large proportion had multiple infections. The study established the occurrence and prevalence of a number of intestinal parasites in the population of Guyana. This sets the stage for the design and implementation of more detailed epidemiological studies. PMID- 12089871 TI - A preliminary study of the indices of obesity among a select group of apparently healthy Caribbean subjects. AB - The purpose of this study was to assess the anthropometric indices of obesity among apparently healthy Caribbean subjects who have the potentials of living an affluent lifestyle. One hundred and eleven (38 males, 73 females) young adults aged between 17 and 38 years participated in the study. Subjects provided information on age, ethnic group, educational attainment, occupation, and previous record of body mass index (BMI) and family history of diabetes mellitus in a self-administered research questionnaire. Waist and hip circumferences (cm), weight (kg) and height (m) were measured. Subsequently, BMI and waist-to-hip ratio (W/H) were calculated. The majority of the subjects (83%) had never measured their BMI. Although the male subjects were significantly taller and heavier than the females (p < 0.01), there was no significant difference in the prevalence rates of obesity between the male and female subjects (p > 0.05). In this preliminary study there was a trend for more females than males to be underweight (BMI < 20 kg/m2). The male subjects had significantly higher waist circumference than the females (p < 0.01), but both genders had a similar percentage of male and female subjects with increased waist circumference (p > 0.05). The limitations of this study are its small size and self selection bias and hence the results obtained must be interpreted with caution. We suggest that assessment of body mass index should be incorporated in the routine clinical measurements of all patients to facilitate early identification, evaluation and treatment of overweight and obesity. PMID- 12089872 TI - Presence of anorexia nervosa and bulimia nervosa in Jamaica. AB - We could find no previous data describing the extent to which eating disorders are a public health problem in Jamaica, and so we carried out two exercises to assess this. We investigated the number of cases of anorexia nervosa (AN) and bulimia nervosa (BN) presenting at the University Hospital of the West Indies (UHWI) between 1985 and 1998, using case records, and carried out a survey among health professionals (psychiatrists, clinical psychologists, paediatricians and nutritionists/dietitians) to determine the number of patients with eating disorders seen by them between 1996 and 1998. We also examined the diagnostic criteria used and correlates of eating disorders. Only two cases of AN were treated at UHWI. Eleven cases each of AN and BN (two males) had presented to the health professionals surveyed, chiefly the psychiatrists. The AN patients ranged in age from 14 to 28 years (mean 20.9 years), and the BN patients from 11 to 35 years (mean 22.2 years). Eating disorders were reported primarily among urban dwellers (76%), and half of the cases were among students. Limiting food intake, excessive exercise and vomiting were the most frequently used methods of weight control. Nine eating disorder patients (41%) were also diagnosed with depression, and five (23%) patients reported previous emotional trauma. The occurrence of eating disorders in Jamaica appears to be very low. PMID- 12089873 TI - Ultrasonography of the lower limbs for deep vein thrombosis at the University Hospital of the West Indies. A five-year analysis. AB - A study was conducted on all patients referred to the ultrasound service of the radiology department of the University Hospital of the West Indies, Kingston, Jamaica, for evaluation of a clinical diagnosis of thrombosis of the deep veins of the lower limb, during the period January 1, 1995, to December 31, 1999. The calf veins were not routinely examined; this study was limited to the popliteal, superficial and deep femoral veins. Two hundred and seventy-four females and 104 males were referred. Seventy females and 32 males had ultrasound findings of deep vein thrombosis (DVT). The significant findings were that the male:female ratio for confirmed DVT was approximately 1:2; 26% of referrals had DVT on ultrasound; DVT was more common on the left and the popliteal vein (48%) and the superficial femoral vein (47%) accounted for most documented cases of DVT but thrombosis of the profunda femoral vein was unusual accounting for only 5% of cases. PMID- 12089874 TI - Analysis of cervico-vaginal (Papanicolaou) smears, in girls 18 years and under. AB - This study was conducted retrospectively at the Queen Elizabeth Hospital and a private laboratory in Barbados to determine the types of epithelial abnormalities in cervico-vaginal Papanicolaou (Pap)-stained smears, and their clinical implications in Barbadian girls, 18 years and under, during the five-year period January 1995 to December 1999. Two hundred and sixty-five Pap smears from 236 patients were examined and the gynaecological history, initial and repeat Pap smear diagnoses, and histology reports of these patients were analyzed. Of the 236 first-visit smears, 94 (39.8%) were abnormal with 36 (15.3%) displaying cytologic features of squamous intra-epithelial lesions (SIL), (33 low grade and 3 high grade). A diagnosis of atypical squamous cells of undetermined significance (ASCUS) was reported in the remaining 58 (24.5%) abnormal smears, of which 35 (60.3%) were suspected to be related to human papillomavirus (HPV) infection. Twenty-two (23.4%) of these 94 patients, who had abnormal smears of either ASCUS or low grade squamous intra-epithelial lesions (LSIL) were re evaluated within six to twelve months of the initial abnormal Pap smear diagnosis. Eight of these 22 patients (36.4%) had histological diagnosis of LSIL inclusive of cervical intra-epithelial neoplasia grade 1 (CIN 1) and condylomata. High-risk HPV DNA types were detected in two of these eight patients (25%). The study confirms that sexually active teenage girls are at risk of developing SIL and high-risk HPV infection. Screening of sexually active teenaged girls by Pap smears followed by other appropriate investigative procedures is recommended. PMID- 12089875 TI - Evaluation of oestrogen and progesterone receptors, and c-erbB-2 in carcinoma of the breast in Trinidadian women. AB - One hundred and three consecutive cases of breast cancer in Trinidadian women were evaluated for steroid receptor status and c-erbB-2 receptor along with conventional parameters including age, ethnicity, tumour size, histological type and grade, and lymph node status: The molecular markers were studied by immunohistochemistry (IHC) on paraffin sections. Tumour size > 2 cm was seen in 60% of the cases. Oestrogen receptor (ER), progesterone receptor (PR) and c-erbB 2 showed 54%, 46% and 63% positivity, respectively. There was no correlation between c-erbB-2 and steroid receptors. Forty-one per cent of cases showed double negativity for steroid receptors (ER-/PR-). No correlation was found between the markers and conventional parameters except for a negative correlation with the tumour grade. The high percentage of c-erbB-2 positivity and the high proportion of steroid receptor negativity suggest a need for studies on adjuvant therapy. Integration of selected markers with conventional parameters could help define subgroups for treatment and prognosis. PMID- 12089876 TI - Prostate cancer at the University Hospital of the West Indies in Jamaica. A clinico-pathological profile at the time of needle biopsy diagnosis. AB - A prospective study of the clinico-pathological features of prostate cancer at the time of needle biopsy diagnosis was conducted in the Department of Pathology at the University Hospital of the West Indies over a one-year period. Ninety cases were diagnosed during this time. The mean age of patients at diagnosis was 72 years and the mean Gleason score of the biopsy, 7.3. Serum prostate-specific antigen levels were markedly elevated in the majority of patients, with a mean value of at least 159.8 ng/ml in the 76 patients for whom this information was available. The significance of these findings and that of other histopathological features examined are discussed, particularly in relation to recognized prognostic factors. It is concluded that the diagnosis of prostate cancer in Jamaica appears to be considerably later than that seen elsewhere. This information should provide support for those who advocate the introduction of a vigorous screening process for prostate cancer in Jamaica. PMID- 12089877 TI - Images and diagnoses. Discoid lupus erythematosus. (Chronic cutaneous lupus erythematosus). PMID- 12089878 TI - Images and diagnoses. Fibrous dysplasia with healed oblique pathological fracture through the mid-shaft of the left femur. PMID- 12089879 TI - Images and diagnoses. Single ventricle. PMID- 12089880 TI - Images and diagnoses. Multiple epiphyseal dysplasia. PMID- 12089881 TI - The initial Trinidad experience with Cine MRI in clinical cardiology. AB - We describe the initial Trinidad experience with Magnetic Resonance Imaging (MRI) and Cine MRI as a diagnostic tool in clinical cardiology. Six patients from the following categories were referred for Cine MRI evaluation: congenital heart disease, valvular heart disease, aortic diseases, cardiomyopathy and intracardiac mass. All patients underwent echocardiography. MRI and Cine MRI were performed on all patients using a Siemens Magnetom 1.0 Tesla MR system at MRI Trinidad and Tobago Ltd. Selected patients underwent Angiography and/or computed tomography (CT) scanning. Clinical data and images of the six patients evaluated are described. MRI and Cine MRI provided excellent anatomical and functional details of the heart and aorta in five patients with dissection of the aorta, aneurysm of the ascending aorta, suspected left ventricular apical thrombus, infiltrative cardiomyopathy and arrhythmogenic right ventricular dysplasia. Technical difficulty was experienced with one patient who had a congenital defect (common atrium). In this study, Cine MRI provided excellent images in all but one patient. This new noninvasive technique enhanced diagnostic capabilities and facilitated management in patients with certain cardiovascular diseases. PMID- 12089882 TI - [Usefulness of osteocalcin and CrossLaps as markers of bone turnover in children with chronic renal failure]. AB - Bone disorders resulting from abnormalities in mineral are common in patients with predialytic stage of chronic renal failure (CRF). Disturbances associated with: phosphate excretions, vitamin D3 metabolism, hypocalcemia, increased parathyroid hormone (PTH) and acid-base disturbances lead to bone pathology know as renal osteodystrophy (RO). Estimation of tempo turnover make possible markers, of which concentration means oneself in urine and in serum. The aim of this study was to estimate the urinary carboxy-terminal telopeptide of type I collagen as a index of bone resorption and osteocalcin as a marker of bone formation in correlation with intact PTH (iPTH) in predialytic children. The study group consisted of 39 children aged 6-17 y. All children were divided into 2 groups: I- 21 predialytic pts; II--18 healthy children. The I group was divided into 2 subgroups: Ia--pts with normal range of iPTH; Ib--pts with higher range of iPTH. All patients were tested: for serum concentrations of: calcium (Ca), inorganic phosphorus (P), iPTH, OST; and for urinary CrossLaps (corrected with urinary creatinine). RESULTS: In subgroup Ib revealed correlation between OST and iPTH (p < 0.05); and between CrossLaps and OST (p = 0.05). CONCLUSIONS: The results of investigation on OST and CrossLaps as bone metabolism markers in predialytic children revealed that evaluating highly specific biochemical parameters of bone turnover is useful in assessing the clinical status of that metabolism. The knowledge of correlation between levels of bone turnover markers and clinical status of a child might help in taking appropriate therapeutic decision and in preventing renal osteodystrophy. PMID- 12089883 TI - [Acute pyelonephritis--clinical picture and the main diagnostic and therapeutic problems in children]. AB - The aim of the study was to determine the main clinical and organising difficulties affecting quality and efficacy of medical care in children with acute pyelonephritis (AP). 41 children aged 3 wk--17 yr 4 mo hospitalized for AP at the Department of Nephrology, District Children's Hospital, Szczecin, Poland in 2000 were investigated. Epidemiologic and demographic data, history, clinical presentation, auxiliary investigations, treatment and ambulatory follow-up were analysed. Instead of typical clinical picture neither diagnostic management nor the therapy fulfilled accepted standards. The main problems found were: delay of referral to hospital by primary care doctor, misdiagnosis or inaccurate primary diagnosis followed by insufficient treatment, difficulties with performing simple ambulatory tests (urinalysis, urine culture, ultrasonography) before hospitalisation and lack of co-operation between parents and nephrological outpatient clinic after discharge. For reflux and obstructive nephropathy, often presenting as urinary tract infection, are still the main cause of chronic renal failure in children in Poland the special attention was paid to necessity of thorough education of family doctors in the subject of management of such cases as well as improvement of accessibility to specialistic care. PMID- 12089884 TI - [Defects of the low urinary tract as a cause of recurrent urinary tract infection in girls]. AB - 462 girls aged 1 month--15 years with recurrent urinary tract infection were diagnosed in the Outpatient Clinic of Mother and Child Institute during 1985-1992 and 1999-2000 years period. The most common cause of recurrent urinary tract infection was an isolated construction of external urethral ostium--59.9% (187/312) then vesico-ureteral reflux coexisting with external urethral ostium- 41.1% (125/312), next isolate vesico-ureteral reflux--22.5% (104/462). Typical radiological diagnostic was performed in all cases and calibration of urethrae was 416 girls. From 75% (312/416) cases with urethral construction only 54.7% (253/462) had typical cystographic picture of that defect. Results of our studies confirmed the importance of exact diagnosis of "lower part of urinary tract" and necessity of appropriate treatment. PMID- 12089885 TI - [Usefulness of Uro-Vaxom in complex treatment of recurrent urinary tract infections in girls]. AB - Uro-Vaxom was used in the treatment of recurrent urinary tract infections in 35 girls. Most of them (34/35) tolerated the drug very well, no side effect were observed. We stopped administration of the Uro-Vaxom in one girl, during the first month of treatment because of vomiting. This way efficiency of Uro-Vaxom was evaluated in the treatment of recurrent urinary tract infections in 34 girls. Uro-Vaxom was found to be a valuable drug, supporting antibiotic therapy in recurrent urinary tract infections caused by E. coli. PMID- 12089886 TI - [Influence of selected factors on the development of renal scars in children with vesico-ureteric complex]. AB - Although relationship between vesico-ureteric reflux, urinary tract infection and renal scarring is known, pathogenesis of the renal scarring is still unclear. Vesico-ureteric reflux and urinary tract infection remain in very close relationship, they may also occur separately. The factor promoting renal injury is also young age of a child. The aim of this paper was to analyse the influence of the selected factors, namely: degree of the vesico-ureteric reflux; child's age at reflux diagnosis time as well as the frequency of recurrences of urinary tract infections exerted on reflux nephropathy. In 45 children with diagnosed vesicoureteric reflux the intensity of renal scarring was estimated using planar scintigraphy with DMSA-Tc-99m and a questionnaire was prepared. The questionnaire included information as follows: age at which the diagnosis was established, degree of reflux at the time of diagnosis, treatment applied, frequency of urinary tract infections before and after having taking medical care of children. It was revealed that the degree of scarring correlated with degree of vesico ureteric reflux of particular kidney. In the investigated group no correlations were found between intensity of renal scarring and both the age of establishing diagnosis and frequency of urinary tract infections before starting the treatment. Taking the obtained results into consideration it seems, that the degree of vesico-ureteric reflux estimated prior to treatment determines degree of renal injuries though, undoubtedly all factors being studied affect renal scarring. PMID- 12089887 TI - [Clinical investigations of children with chronic renal failure and peritonitis treated with peritoneal dialysis]. AB - Peritonitis and catheter-related infection are the most-common complications of peritoneal dialysis treatment. The aim of study was to analyse frequency causes, effectiveness of therapy and risk factors of peritonitis in patients treated with continuous peritoneal dialysis (CAPD). We described 13 patients aged 6-21 years, mean 12 years, who initiated CAPD between 1995 to 2000. The most-common primary cause of end-stage renal disease were chronic glomerulonephritis in 8 children, renal dysplasia in 3 children, hemolytic-uremic syndrome in 1 child and obstruction uropathy in 1 child. Peritonitis rate was 1 episode for 9 patient months. Staphylococcus aureus was the most-common infection factor. We performed exchange of catheter in 8 children. Causes of exchange were exit site infection, cuff excision, tunnel infection and peritonitis. Children were usually treated with two antibiotics for two weeks. Effect of treatment was better in these episodes treated longer than two weeks. As a risk factors of increasing peritonitis relapses we suggest immunosuppressive therapy, protein deficiency, catheter dislocation, not adequate education of patient family and chronic skin infections. We observed lower rate of peritonitis relapses in patients treated longer than two years on CAPD. CONCLUSIONS: 1. In patients with chronic renal failure treated with CAPD the preceding immunosupressive therapy increases the frequency of peritonitis. 2. More intensive treatment can cause decreasing frequency of peritonitis relapses. 3. Some risk factors increase the frequency of peritonitis relapses. 4. A better education of patient and his family decreases the frequency of peritonitis relapses. PMID- 12089888 TI - [Lower urinary track function in children with urinary track infection]. AB - During the last decade more and more urodynamic studies in children have been performed as a step in diagnostic process of urinary tract diseases in children. Such a procedure is done to diagnose a cause of urinary tract infection as well. The aim of our study was to evaluate uroflowmetric parameters in children with urinary tract infections and try to find typical sings. Among the children examined at the Department as an out-patient we randomly selected the group of 379 patients. There were 94 patients afflicted with urinary tract infection without any special predisposing factors. The range of age was 5-15 years (ave- 9.1, Me--9). There were 84 girls and 10 boys. A control group consisted of 195 patients in the range of age 5-16 years (ave 9.6). We analysed standard parameters of the uroflowmetric protocol (maximal and average flow, urinary bladder volume, residual volume, and time flow). Results were presented as average (X) and standard deviation (SD). There was no statistically significant difference between both group (p < 0.05). PMID- 12089889 TI - [Serum zinc concentration with reference to other markers of continuous ambulatory peritoneal dialysis patients status]. AB - The aim of our studies included measurement of serum Zn level in CAPD patients with the subsequent evaluation of relations between serum Zn and markers of nutrition, dietary intake, markers of acute phase reaction, CAPD adequacy, nitrogen balance as well as routine clinical and laboratory data. The study was performed in 81 patients treated with CAPD for up to 3 years (12 3-month study periods). Mean serum Zn concentration was 12.2 +/- 1.8 mumol/l and was decreased in 16% of patients. Positive correlation was shown between serum Zn level and prealbumin, iron, transferrin saturation, haemoglobin, mean corpuscular haemoglobin concentration and dialysis duration. Negative correlation was shown between serum Zn level and patients age as well as daily influent and effluent volumes, what means that patients in age over 65 years and those using daily inflow or outflow dialysate volume greater than 12.7 and 12.9 I, respectively, are at risk of Zn deficiency. Dietary Zn intake (9.9 +/- 2.5 mg/day) was in 96% of patients lower than that recommended for CAPD patients. Relation between Zn intake and ferritin, total cholesterol and vitamin E in serum as well as HDL /total cholesterol ratio was shown. Our results indicate that approx. 16% of CAPD patients need Zn supplementation. Serum Zn level in CAPD patients shows a beneficial effect on serum markers of iron metabolism, blood morphology indices, serum lipid profile and increased serum vitamin E level. PMID- 12089890 TI - [Dialysis-related complications in peritoneal dialysis patients]. AB - The hospital records of 56 patients (25M, 31F) with acute or chronic renal failure treated by peritoneal dialysis were retrospectively reviewed. Mean dialysis time was 13 +/- 15 days in acute renal failure and 32 +/- 23 months in chronic renal failure. The incidence of infectious (exit site infections and peritonitis) and non-infectious dialysis-related complication was assessed. Exit site infections were significantly more frequent in children aged 5 or less than in older patients (1/9.6 patient-month and 1/26.5 patient-month, respectively, p < 0.001). Such relationship was not found with regard to the incidence of peritonitis. There was a tendency of peritonitis rate to decrease in consecutive years was noted. This can be probably related to an increase in the number of patients, introduction of automated peritoneal dialysis, increasing experience of medical staff and patients themselves. PMID- 12089892 TI - [Overweight and obesity in urban population in Poland in years 1983-1999]. AB - There is a shortage of up-to-date data on the extent of the problem of overweight and obesity in Poland, although obesity is commonly considered as the 'epidemic of the end of the 20th century'. The aim of the study was to compare the percentages of overweight (BMI > or = 25) and obese (BMI > or = 30) adults from Wroclaw population in particular categories of age, sex and social status. Material comprised the data of 15,641 men and 19,121 women aged 21-60, occupationally active inhabitants of Wroclaw, examined in the DCDM 'DOLMED' in 1983-1999. BMI was used as a measure of general obesity (according to WHO categories). It was revealed that age, sex and social status significantly differentiated BMI of examined inhabitants of Wroclaw. In the subsequent decades of life mean BMI values increased, hence an increase of percentages of overweight and obese persons was observed. More than the half of men exceeded their proper relative mass before 40 years of age, whereas the 2/3 of them had BMI over 25 before the age of 50 (independently on their social background). The process of increasing percentages of overweight women was socially differentiated; between the 3rd and the 6th decade of life the percentages of overweight women increased among intelligence from 16 to 60%, among clerks from 18 to 72% and among workers from 27 to 83%. A sexual dimorphism resulted in fact that in men an age-related increase of overweight subjects (25 > or = BMI < 30) was observed, whereas in women an age-related increase of obese persons (BMI ? 30) was found. In an urban population in Poland in the 1990s the percentage of overweight persons (BMI l 25) exceeded 70% after the age of 50. Therefore--in the context of essential relationships between overweight and both mortality and morbidity--the health status of urban population in Poland is highly disturbing. PMID- 12089891 TI - [Captopril in the treatment of acute myocardial infarction. Hypotension as an important clinical problem]. AB - The aim of study was to analyze the effect of captopril (C) on blood pressure in patients with acute myocardial infarction (AMI) during 1 year treatment. Patients less then 70 years old with systolic blood pressure (sBP) > or = 100 mm Hg were qualified to the study. Administration of C was started during the first 4 days of AMI (mean 21 +/- 24 h). 50 pts treated with C and control group of 43 pts were finally analyzed. Doses of C were gradually increased from 3.125 t.i.d. on the first day till 25 mg t.i.d. on the 4th day. Significant decrease of sBP after administration of C was observed at 60 and 120 min after the 1st, 2nd and 3rd dose on the first day, and after 30-120 min after first C dose on the 4th day. dBP decreased only at 60-120 min after the 1st dose on first day and at 90 min after the first dose on 4th day. Hypotension after 1st and 2nd dose of C on the first day caused exclusion of 3 pts (6%) from the study. In conclusion, hypotension seems to be quite often encountered during captopril therapy of early phase of AMI. It appears not only after the first but also after the following doses. The initial dose of 3.125 mg seems to be safe and sufficient to assess its hypotensive action. PMID- 12089893 TI - [Prenatal diagnosis of congenital anomalies requiring surgical management in the Lower Silesia--fact or fiction?]. AB - In order to assess the current status of prenatal diagnosis of congenital anomalies requiring surgical management in the area of Lower Silesia, a retrospective study of medical charts of all neonates admitted to the Department of Paediatric Surgery in Wroclaw between 1997 and 2000 was undertaken. 218 babies affected by anomalies detectable antenatally by conventional ultrasound scan were subjected to the further study. Neural tube defects and abdominal wall defects were the most common and were diagnosed in 63 and 38 children respectively. The incidence of other congenital anomalies; hydrocephalus, oesophageal, duodenal and intestinal atresia, diaphragmatic hernia and urinary tract anomalies was similar in each group. The suspicion of anatomical defect or its precise recognition was made antenatally in 52 pregnancies (23.8%). The highest rate of positive prenatal diagnosis was noted among babies with hydrocephalus (50%) and urinary tract anomalies (42.8%). In other groups this rate was significantly less. Critical analysis of postnatal management in studied cases revealed that the babies with atresia of gastrointestinal tract, congenital diaphragmatic hernia and abdominal wall defects were negatively affected by a lack of prenatal recognition of the defect. Moreover, in more than 40% of all analysed pregnancies presence of risk factors (positive family history, course of pregnancy, mother's age, polyhydramnios, oligohydramnios, previous abnormal pregnancies) were identified that should have prompted an obstetrician to a through diagnostic evaluation of the foetus. The obtained results clearly indicate the necessity for the elaboration of a new protocol of antenatal care of pregnant women in the Lower Silesia. In case of suspected anomaly amenable to operative correction within the first days of life, paediatric surgeon should be an active member of a specialist team responsible for care of mother and her baby. PMID- 12089894 TI - [Pneumoperitoneum and peritonitis in a child treated by continuous ambulatory peritoneal dialysis]. AB - Pneumoperitoneum is a rare complication of peritoneal dialysis, which in most cases occurs soon after the implantation of the peritoneal catheter and does not need any specific treatment. In contrast, pneumoperitoneum due to visceral perforation represents a serious clinical problem and usually needs an urgent surgical intervention. We present a case of a 10-year old girl treated by peritoneal dialysis for 7 months who was admitted to hospital with symptoms of peritonitis and pneumoperitoneum. On admission her condition was severe and visceral perforation was strongly suspected. As her parents did not give consent to surgical treatment, only conservative management was introduced. Peritoneal dialysis was continued and the girl responded well to antibacterial therapy. The final outcome was favourable. PMID- 12089896 TI - [Causes of disturbances in iron turnover in chronic renal failure]. AB - In many patients with chronic renal insufficiency a depletion of stored and cellular iron (absolute iron deficiency), a blockade of iron in body stores (functional iron deficiency) or an iron overload can be shown. The factors leading to absolute iron deficiency are: 1) loss of iron as a result of blood loss by the gastrointestinal tract, taking blood specimens for laboratory tests and related to dialytic procedure; 2) enhanced use of iron during intensive erythropoiesis stimulated by recombinant human erythropoietin; 3) dietary iron deficiency or impaired iron uptake from the gastrointestinal tract; 4) other forms of gastrointestinal tract impairment; 5) pharmaceutical substances forming inabsorbable iron complexes and/or diluting the acidity of the gastric juice; 6) certain demographic factors. Functional iron deficiency develops during treatment with recombinant human erythropoietin and in the infectious state and during the inflammatory process. The use of recombinant human erythropoietin accelerates erythropoesis and by so increases iron requirement frequently far higher than the ability of iron stores to "transfer" iron to the bone marrow--this may be a result of ineffective mobilisation of iron stores and/or ineffective transport. In the infectious process the use of iron is impaired because of augmented cytokines production which leads to increase iron uptake and storage in the reticulo-endothelial system. Iron overload is caused by excessive iron intake, e.g. parenteral iron administration or repeated blood transfusions. PMID- 12089895 TI - [Vasculitis of cerebral vessels, probable cause of neurological complications in a child with Schonlein-Henoch purpura]. AB - We report 6-years old boy with Schonlein-Henoch purpura who presented neurologic manifestations: depressed state generalised convulsions, and cortical blindness. Sequential magnetic resonance imaging showed bilateral cerebral ischemic lesions in the cortex and white matter of parieto-occipital lobes caused by vasculitis. PMID- 12089897 TI - [Fungal allergy--part II]. AB - Allergic reactions caused by fungi, may be present in the airways, skin, eyes, gastrointestinal tract, skin and to cause systemic reaction. Nasal symptoms react as allergic rhinitis. In a small group of patients the allergic fungal rhinosinusitis is present. It is important, because of the way of treatment, to differentiate the AFS from the other forms of chronic rhinosinusitis. The fungi may cause bronchial asthma, allergic bronchopulmonary aspergillosis and allergic alveolitis (hypersensitivity pneumonitis). Diagnostics of the allergic reactions caused by fungi includes routine allergological methods, mycologic culture and direct microscopic examination. The process of reaching the diagnosis of allergic fungal rhinosinusitis and allergic bronchopulmonary aspergillosis may require the CT and histopatologic examination. The treatment of fungal allergic reactions includes allergen avoidance, pharmacotherapy, antifungal treatment (id reactions) and surgical treatment (allergic fungal rhinosinusitis). The immunotherapy is a valuable, and accepted by the WHO therapy of the Cladosporium and Alternaria sensitive patients. PMID- 12089898 TI - [Mycoplasmal infections--still actual problem]. AB - In the research there was presented a wide variety of infections caused by different Mycoplasma subtypes, the most frequently systemic location, diagnostic and treatment procedures and problems connected with. PMID- 12089899 TI - [Selected problems of endothelial functions. I. The role of endothelium in maintaining the hematological and circulatory balance]. AB - Based on literary data we carried out the analysis of the influence vascular endothelium on maintaining the haematological and circulatory balance in the organism. Particular attention was paid on its active metabolic and secretory role in this process. PMID- 12089900 TI - [Selected problems of endothelial functions. II. The role of the selectines in the damage of vascular endothelium]. AB - Studies in the recent years have provided numerous evidence of the inflammatory and immunological character of atherosclerosis. In the early stages of the adhesion of leucocytes and platelets to the blood vessel wall, and also in occurring intercellular interactions leading to creating the atherosclerotic lamina, the adhesive molecules from the selectine family ply the crucial role. Their expression, mainly selectines L, P and E on the endothelial surface and on the smooth muscles of the vessels, undergoes a distinct intensification under the influence of hypoxia. The begun cascade of molecular changes with the contribution of selectines leads inevitably to the development of atherosclerotic changes. The neutrophile and lymphocyte fixed to the endothelium show ability to its damage, both as a result of immediate response [release of free oxygen radicals, enzymes and cytoxic substances] and as a result of indirect action [release of cytocines which change the biological property of the endothelium- the so called endothelial activation]. In this work the role of diabetes and hypertension in the intensification of those processes has been noticed. PMID- 12089901 TI - [Endothelial dysfunction in hypertension--clinical implications]. AB - Endothelial cells produce both vasodilatating compounds as nitric oxide, prostacycline, endothelial derived hyperpolarising factor and counteracting substances known as endothelial derived contracting factors: endothelin, tromboxan A2, prostaglandin H2, free oxygen radicals. Natural balance between both groups affects blood perfusion of various tissues and constitutes important element in blood pressure control. More and more attention is paid to endothelial dysfunction in patogenesis of hypertension. In a number of studies endothelial dysfunction in hypertensive patients was found out as decreased release of nitric oxide or increased production of endothelin. Principle mechanism of impaired function of endothelium in hypertension seems to be decreased production and increased degradation of nitric oxide mainly due to free oxygen radicals. Favorable effects in improvement of endothelial function were achieved by using ACE inhibitors, AT1 receptor blockers and calcium channel antagonists. PMID- 12089902 TI - [Programmed cell death in the pathogenesis of human diseases]. AB - The distempers considering cell dying have an important meaning in pathogenesis of different diseases, just like the proliferation distempers. Proliferation and dying are probably responsible for homeostasis support at the same part. Two groups of distempers were distinguished. One of them is connected with an increased cell viability (some cancers, autoimmunological diseases, viral infections) and the other where there is an increased cell mortality (AIDS, some neurodegenerative diseases, myelodysplastic syndromes, osteoporosis and others). Programmed cell death induction using gene therapy methods is tested in preclinical examinations. The results of these tests are thought to have an important significance in therapy of many diseases. PMID- 12089903 TI - [Interaction of 5-HT and HPA axis in depression and treatment-resistant depression]. AB - Psychoendocrinology studies of depressed patients focus on the disregulation of the hypothalamic-pituitary-adrenal (HPA) axis. Abnormalities in the HPA axis have been noted in depressed patients. Numerous data have demonstrated the existence of reciprocal interactions between the central serotonin (5-HT) system and HPA axis. These interactions are of particular relevance when considering pathological conditions, such as depression, in which modifications of both the 5 HT system and HPA axis have been evidenced. In our laboratory, we examined the effects of adrenocorticotropic hormone (ACTH) on the immobilization of rats in the forced swim test and on the wet-dog shakes induced by the DOI, 5-HT2 receptor agonist with the administration of imipramine and lithium. The reduction of immobility, induced by the chronic administration of imipramine for 15 days, was blocked by treatment with ACTH for 14 days. And, chronic ACTH treatment for 14 days increased the wet-dog shake response. This effect of ACTH was not inhibited by a 14-day administration of imipramine. Accordingly, the chronic treatment of rats with ACTH may prove to be an effective model for antidepressant-treatment resistant depression. We believe that behavioral pharmacological and molecular biological research into the interaction between the 5-HT and HPA axis will elucidate the pathogenesis of depression or antidepressant-treatment-resistant depression and the mechanism of antidepressants action. PMID- 12089904 TI - [Analysis of neuronal functions in mice lacking the NMDA receptor epsilon 1 subunit]. AB - The NMDA subtype of glutamate receptor (GluR) plays an important role in excitatory neurotransmission, synaptic plasticity, brain development, and neurodegeneration. NMDA receptors are inherent high Ca(2+)-permeable channels, which are formed by heteromeric assembly of the GluR zeta 1 subunit (NR1) and any one of four GluR epsilon subunits (GluR epsilon 1-4; NR2A-D). Mice lacking the GluR epsilon 1 subunit exhibited a malfunction of NMDA receptors, as evidenced by reduction of NMDA receptor channel current, hippocampal long-term potentiation, [3H]MK-801 binding, and NMDA-stimulated 45Ca2+ uptake. A biochemical analysis revealed a hyperfunction of dopaminergic and serotonergic neuronal systems in the frontal cortex and striatum of GluR epsilon 1 mutant mice. The enhancement of dopaminergic neuronal activity in the striatum, at least, due to the disinhibition of inhibitory GABAergic neuronal input. GluR epsilon 1 mutant mice showed an increase of locomotor activity in a novel environment attributed to the hyperfunction of the dopaminergic neuronal system, and an impairment of spatial, contextual, and latent learning. These findings provide evidence that NMDA receptors regulate behavior through the modulation of not only glutamatergic but also GABAergic and dopaminergic neuronal systems. Moreover, it is suggested that GluR epsilon 1 mutant mice are useful as an animal model, which is associated with the malfunction of NMDA receptors and hyperfunction of the dopaminergic neuronal system. PMID- 12089905 TI - [Introduction of the conventional method of DNA transfection by adenovirus vector]. AB - DNA transfection techniques are necessary to perform the high throughput screening (HTS) for drug discovery. Among many transfection techniques, we will recommend the adenovirus vector for HTS as a particularly useful method. The adenovirus vector has attracted a great deal of attention as a vector for gene therapy and is also useful for studying gene functions in differentiated cells such as neurons and muscle cells. An efficient method of constructing recombinant adenoviruses has been established. In this article, we will introduce the method to produce the desired recombinant adenoviruses and show our findings using these adenoviruses. PMID- 12089906 TI - [Evaluation of pro-arrhythmic risk of drugs due to QT interval prolongation by the HERG expression system]. AB - Recently, there has been considerable attention focused on drugs that prolong the QT interval of the electrocardiogram. This occasionally evolves to fetal, polymorphic ventricular arrhythmias, torsades de pointes. Therefore, the early detection of the risk of drug-induced QT prolongation is important for avoiding the adverse cardiovascular effect in clinical use. It has been suggested that the QT prolongation and ventricular arrhythmia caused by drugs might be secondary to their ability to interfere with cardiac potassium channels involved in action potential repolarization and in particular with rapidly activating delayed rectifier K+ current (IKr). In cardiac myocytes, IKr contributes to termination of the plateau phase of action potential. The ether-a-go-go related gene in humans expressed a K+ channel current with biophysical characteristics similar to those of IKr. Electrophysiological studies on cloned HERG channels can provide fundamental information concerning the cardiac safety profile of new developing drugs. PMID- 12089907 TI - [The role of anthropologic study in the stomatognatic morphologic changes through time]. AB - OBJECTIVES: The possibility of a prognosis regarding the morphologic aspect of the passive component of stomatognathic apparatus was assessed through some researches on the viscero-cranium aspect. METHOD: The study used 20 skulls or skull fragments, 8 were from the first century A.D., 4 from the Middle Ages and the rest of 8 were from the XX-th century. All The skulls were found on the Romanian territory. A series of measurements were made on the maxilla, mandible and on the face as a whole; the results were compared with the literature data. RESULTS: The skulls discovered in the Neolithic necropolis existing on the Romanian territory are of mediterranoic type, with an antero-posterior prolonged skull and with narrow dental arches, presenting a prognathism according to the shape of the skull. After the Neolithic, the shape of the skull changes to a more rounded one and the dental arches become upsiloid. In the Middle Age, the dental arches tend to evolve from the upsiloid type to the paraboloid type. In our century, the paraboloid form of the dental arches is maintained (semiellipitic shape of the maxilla and the parabolic shape of the mandible). CONCLUSIONS: During the ages, the evolution of masticator apparatus was influenced mainly by the genetic factor regarding the shape and the pathology, but also, in some degree, by the dietary habits. Multiple changes have been made possible a gene crossing between individuals. Due to the genetic mixture between the local element and the migratory population, the evolution of the stomatognathic apparatus to paraboloid dental arches is explained, although upsiloid dental arches can be found. PMID- 12089908 TI - [Assessment of daily aluminum intake by food consumption]. AB - Aluminum is naturally present in foods; the contents of aluminum may increase during cooking or technological processes, or during conditioning. The average aluminum intake for humans is 10 mg/day. In case of drug administration, the normal average intake may reach 50-1000 mg/day. Foods represent the major source of aluminum for humans. We assessed the aluminum contents in samples of vegetal (vegetables and fruits), animal foods (milk and dairy, meat ) and drinking water, in order to evaluate the daily aluminum intake. Aluminum assessment was made by the colorimetric method with eriocromcianin R. The gathered values (mg Al3+/100 g product), range between 0.0177 and 0.5473, are similar to those reported in the literature, and allow the assessment of aluminum intake by daily food consumption. The daily aluminum intake by means of normo- and hypocaloric menus was computed, and ranges between 2.4231 and 0.8677 mg aluminum. PMID- 12089909 TI - Treatment of achalasia: an update. PMID- 12089910 TI - [Pathology of dermatoglyphics in ocular diseases]. AB - The paper studies dermatoglyphics, viewed from a pathological perspective, on a group of 200 blind subjects or suffering from other grave congenital ocular affections, many of them hereditary, living in the province of Moldavia, from which 400 finger prints and palms have been collected. The observation to be made in that the advanced degree of sight's affection, up to the total loss, is correlated with an ample and strong pathological charge of the digital and palmar dermatoglyphic image of the subjects affected, suggestively expressed at the sample's level by a higher incidence of the distortions with serious medical implications, quite close to that recorded at deaf-mutes, mental handicapped people, in the grave congenital cardio-vascular affections, or at parents with malformed children from Moldavia. From the multitude of dermatoglyphic distortions put into evidence, the author considers that it is only one that might be specific to the serious congenital grave ocular affections, that is a certain disposal of the epidermal ridges, as a dense or very dense network at the level of the Th/I, in a ratio of 80% with women and 38% with men. The very high frequency of this distortion for females subjects might assume that the gene responsible for such a pathological configuration is placed on chromosome X. PMID- 12089911 TI - [Intestinal occlusion by persistence of the omphalo-enteric conduct in a 10-year old child]. AB - The persistence of the omphalo-enteric conduct is a rare, but possible, cause of pathology in children. Haemorrhage and intestinal occlusion are the most frequent clinical presentations, usually in infants. We describe the case of a 10 year old boy, who was admitted in our department for what seemed first to be an acute appendicitis. During hospitalization, the clinical picture turned on to an intestinal occlusion, and the patient finally required surgery. We performed exploratory laparotomy and we discovered a mechanical occlusion by persistence of the omphalo-enteric conduct, which was treated according to standard protocols. The post-operative course was good, and the child was discharged 8 days after surgery. This case emphasizes that the persistence of the omphalo-enteric conduct should be sought as the source of symptoms in any child presenting with intestinal occlusion. PMID- 12089912 TI - [Hemorrhagic risk factors during beta-lactam antibiotics therapy]. AB - Any type hemorrhagic manifestation may occur 12 hours to 5 weeks after the administration of beta-lactam antibiotics. The mechanisms of blood losses proved to be by: 1) immunologic thrombocytopenia (penicillins); 2) alteration of platelet functions (semisynthetic penicillins); 3) hypoprothrombinemia (cephalosporins). The risk factors for the occurrence of hemorrhage under beta lactam antibiotics therapy are: concomitant administration of cytostatics for a neoplastic malignancy; b) acute or chronic renal failure; c) concomitant treatment for duodenal and gastric ulcer; d) malnutrition; e) dosage and duration of antibiotic administration. The frequency of bleeding under beta-lactams is not determined as yet. A severe case diagnosed at the IIIrd Medical Clinic of Iasi presenting spontaneous daily nasal bleedings that occurred 24 hours after the initiation of the treatment with cephalosporins (Kefurox) is presented. In this patient the risk factor was chronic renal failure. PMID- 12089913 TI - [Goldenhar syndrome in adults]. AB - The paper presents the case of a 43 year old female patient with multi malformations syndrome (facio-auriculo-vertebral syndrome), distinguished by alterations of the skeleton, face, sensorial organs and the heart. The syndrome is dominant at young ages and scarce at adults, and the explanation of the favorable evolution could be the lack of severe visceral or central nervous system involvement. The peculiar anomalies of this case are: asymmetric face, epibulbar dermoid, dysplastic ears, auricular tags, conductive and sensorineural deafness, fusion of vertebrae, hemivertebrae, ventricular septal defect. PMID- 12089914 TI - [Castleman's disease. Discussion related to case report]. AB - Castleman's disease, also called angiofollicular hyperplasia was first described as a distinct entity by Castleman et al in 1956. Now there are described two forms, a localised and a multicentric one. All forms of disease share the same histopathological features with a highly characteristic angiofollicular lymphoid hyperplasia which is either hyalinovascular or plasmocytic. The clinical and biological signs are varied and heterogeneous. The disorder is of unknown origin, but interleukin 6 plays a central part in this disease. Despite the benignity of this "prelymphoma state", an aggressive course with poor prognosis occur usually in the multicentric form. We report a case of Castleman's disease, multicentric variant. This case provides the opportunity for discussing many aspects of this atypical lymphoproliferative disorder. PMID- 12089915 TI - [Scintigraphy in breast cancer]. AB - Scintigraphy was evaluated in comparison to clinical and paraclinical methods used in diagnosis of breast tumors. 32 patients with palpable breast tumors were investigated by clinical examination, ultrasound, mammography and scintigraphy. Different radiopharmaceuticals marked with 99mTc were used, having different affinity for the tumoral tissue. A Siemens Diacam gamma camera was used. Patients were submitted to dynamic study of early blood-pool and tissular phase, followed by late static incidences. Regions of interest were compared with similar areas of the contralateral breast. Conclusions were compared to histopathological diagnosis. We have calculated sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for each of the investigation methods. The sensitivity was 80% for clinical examination, 100% for ultrasonography, 66% for mammography and 100% for scintigraphy. The specificity was 77% for clinical examination, 50% for ultrasonography, 90% for mammography and 80% for scintigraphy. PPV was 75% for clinical examination, 60% for ultrasonography, 80% for mammography and 82% for mammography. NPV was 82% for clinical examination, 100% for ultrasonography, 71% for mammography and 100% for scintigraphy. Good correlation was found between scintigraphy and histopathology. Ultrasound and mammography also confirmed, in most cases, the clinically suspected diagnosis. Including scintigraphy in the investigation protocol for breast tumors will allow a better assessment of the therapy. PMID- 12089916 TI - [Physiology 2000--retrospect and perspectives]. PMID- 12089917 TI - [The use of high gradient magnetic separation (HGMS) in obtaining red blood cell mass]. AB - This paper presents the "high gradient magnetic separation" (HGMS) and its use in red blood cells separation. A high gradient magnetic field system is described (set up from a filamentary iron-magnetic matrix), that is specially design for red blood cells capture. The configurations and the conditions that are needed for the red blood cells concentration process to allow the red blood cells mass to by obtained are evaluated. Finally, another possible technique of red blood cells concentration based on a type of continual magnetic separation is described. PMID- 12089918 TI - [History of Romanian Surgery]. PMID- 12089919 TI - [Professor Matei Bals (1905-1989 ) - Friend and fond of Iasi]. PMID- 12089920 TI - The extra-esophageal manifestations of gastroesophageal reflux disease: pathophysiology, diagnostics and treatment. PMID- 12089921 TI - [Cellular electric potentials in experimental cardiac hypertrophy]. AB - Myocardial hypertrophy is considered as an adaptive mechanism of the heart submitted to mechanical overload manifested by an increase of ventricular mass. For the study of the myocardial hypertrophy different experimental models were proposed although the transposition of the results in clinical conditions is difficult. In the adaptive mechanisms of the heart of great importance are the electro-mechanical interrelations. Mechanical alterations during cardiac hypertrophy are characterized by a reduced shortening velocity and an increase of the time to peak tension. They are due to the prolongation of action potential duration especially of its plateau and a decreased rate of repolarization because of the increase of the peak ICa and ICa density, and the slower inactivations of the maximal ICa. Abnormalities of the K+ currents IK1 and IK may contribute to the prolongation of action potential duration. The instability of Ca2+ transients in hypertrophic myocardium can cause the apparition of cardiac arrhythmias. PMID- 12089922 TI - [Renal osteodystrophy (I)]. AB - Renal bone disease represents one of the major complications of end-stage renal disease, accounting for the numerous and various changes at bone level, determined by abnormal calcium and phosphorus homeostasis and by changes in calcitriol and PTH synthesis. PTH represents as well a major uraemic toxin, exerting profound systemic effects, particularly at the cardiovascular level. PTH synthesis is mainly controlled by changes in calcium-phosphorus balance and calcitriol production by the kidneys. Several others factors are important in the development of secondary hyperparathyroidism: acidosis, autonomisation of PTH secretion and peripheral (target-organ) resistance to PTH actions. Although bone biopsy represents the definitive diagnostic test to differentiate between osteitis fibrosa, low-turnover bone disease and bone involvement unrelated to disturbed calcium metabolism (i.e. beta 2-microglobulin-related amyloidosis), plasma intact PTH generally exhibits a reasonably good relation with bone histology parameters. Moreover serum bone-specific alkaline phosphatase isoenzyme, serum pyridinoline and the novel serum markers for bone turnover are highly specific and correlate with bone histomorphometry parameters, so that, preventive and therapeutic strategies should be re-evaluated based solely on biochemical parameters. PMID- 12089923 TI - [Actual considerations in pneumococcal meningitis]. AB - Streptococcus pneumoniae has been recognized as a bacterial pathogen in human beings. It was the main cause of meningitis, almost since it was first isolated by Sternberg and Pasteur more than 100 years ago. Meningitis is constantly lethal without specifically treatment. Emergence and diffusion strains of the Streptococcus pneumoniae indicate the changes that are likely in the area of antimicrobial resistance in the last decade. Resistance of Streptococcus pneumoniae to antibiotics has progressed rapidly and is a global problem. This article presents the new information about Streptococcus pneumoniae, and the new possibilities of treatment of the pneumococcal meningitis. PMID- 12089924 TI - [The importance of ano-rectal manometry in irritable bowel syndrome]. AB - Irritable Bowel Syndrome (IBS) represents a frequent cause for gastroenterological referral. 50% of all gastroenterological consultations are for functional gastrointestinal disorders (IBS included). Multiple motility abnormalities were described in IBS. The AIM of the study was to evaluate the role of ano-rectal manometry in the diagnosis of IBS. STUDY DESIGN: 24 patients with IBS (mean age 50.8 +/- 15.4) and a control group of 10 subjects with no abdominal symptoms (mean age 48.1 +/- 12.3) had each an ano-rectal manometry examination. RESULTS: After exclusion of constipation predominant IBS subjects, the IBS patients presented lower perception threshold (22.1 +/- 10.9 ml) than the control group (60.2 +/- 11.3 ml) (p < 0.05) and than the constipation predominant group (81.3 +/- 44.0) (p < 0.05). In 44% of constipation predominant IBS concomitant pathology, revealed by manometry, was present: megarectum or hemorrhoids (spasm). CONCLUSION: Anorectal manometry could be a useful tool for the evaluation of IBS patients. PMID- 12089925 TI - [The prevalence of Helicobacter pylori infection in gastric cancer and gastric stump cancer]. AB - In 1994, the International Agency for Research in Cancer recognized Helicobacter pylori (Hp) as "a definitive cause of human cancer". The present study aimed at determining the prevalence of Hp infection in the gastric cancer developed on the background of a disordered stomach as compared with gastric stump cancer. The prevalence of Hp infection was a twice higher in the patients with gastric cancer than in those with gastric stump cancer, and in both cases much lower than that reported in the literature. Possible explanation would be the much older mean age of the patients, the protective role of gastrectomy, and the multifactorial etiopathogenesis in both gastric and gastric stump cancers. PMID- 12089927 TI - [The metabolic effects of the angiotensin converting enzyme inhibitors in experimental diabetes mellitus]. AB - We have studied the effects exerted by the angiotensin converting enzyme inhibitor (ACEI) Enalapril upon the glycemic profile in diabetic rats. With respect to the subacute character of our experiment (15 days), the following parameters have been recorded: diuresis, glycosuria, ketonuria and glycemia. In the diabetic rats treated with ACEI we haven noted slight, statistically insignificant improvements in glycosuria and glycemia, compared to the untreated diabetic subjects. Conversely, a statistically significant improvement of ketonuria has been noted in the diabetic rats receiving ACEI compared to the untreated ones. The short-term administration of ACEI in experimental diabetes mellitus acutely improves the glycemic profile, probably by the amelioration of the tissular sensitivity to insulin. PMID- 12089926 TI - [Heart blocks in gallstone]. AB - In this study, I analysed the AV blocks and BB blocks which I found in gallstone. OBJECTIVES: The objectives were to find if the heart blocks are in relationship with gallstone or not. METHODS: From 1122 patients with gallstone--13 had the AV blocks and 95 had BB blocks. All of these were examined: physical examinations, abdominal echography and Ecg. RESULTS: This study, 1.15% from the patients had AV blocks and 6.68% had BB blocks, especially RBB blocks, LBB blocks and AV1 block. CONCLUSIONS: I approximate that these diseases of the heart can coexist with gallstone. PMID- 12089929 TI - [Human Genome Project--medical implications]. PMID- 12089928 TI - [Is the age a limiting factor of functional capacity and cardiac performance in patients with chronic myocardial infarction ?]. AB - Actually the old age is not considered a contra-indication of cardiac rehabilitation, although it influences the prognosis of patients with chronic myocardial infarction. The objective of the paper is the study of age as a limiting factor of the cardiac performance and exercise capacity at patients with myocardial infarction at the admission in rehabilitation programmes. METHODS: Among the patients admitted in the Department of Internal Medicine of Rehabilitation Hospital of Iasi there were selected 132 with chronic myocardial infarction; they were divided into two groups: group A of 56 patients more than 55 years old and group B of 76 under 55. There were analysed the clinical features (risk factors, symptoms), the cardiac performance (by echographic study of the regional and global abnormalities of myocardial contractility) and the functional capacity (by exercise testing at the ergometric bicycle). RESULTS: Both groups were similar by the point of view of the prevalence of the risk factors and location of the infarction. The ejection fraction, although smaller at the group A (45% vs 50%), did not reach the statistical significance. The exercise testing revealed a lower capacity of the group A, marked by higher levels of the cardiac rate and time-tension index at similar levels of exercise. The metabolic equivalent (MET) and VO2max were significantly lower (p = 0.001 respectively 0.002). There can be concluded that the age diminishes the exercise capacity of the patients with myocardial infarction without a marked influence of the cardiac performance. The last one depends on the infarction size and the myocardial status while the lowering of the functional capacity is influenced by noncardiac factors as well. PMID- 12089930 TI - [Risk factors in child and adolescent with systemic hypertension]. AB - A prospective study was made on 1068 pupils, aged between 10-15 years (group I). From this group 995 pupils were examined after four to seven years (group II). Their age was between 16-19 years. The study contained: anthropometric data, blood pressure (BP) and pulse measurements, and a standard record using the epidemiologic interview method. For each pupil we determined the following familial and personal antecedent of cardiovascular disease, especially arterial hypertension; alimentary habits (salt and hydro-carbonate excess); consumption of coffee and cigarettes; physical activity; tendency to sedentarism; school or family stress; and behaviour type. Data were worked out and interpreted, using epidemiologic and statistic methods in Epi Info 6 program. The prevalence of genetic factors (4% in group I and 4.3% in group II), consumption of salt (38% in group I and 35% in group II), obesity (1.5% in group and 1.4% in group II), sedentariness (4.8% in group I and 6.8% in group II), stress (9.8% in group I and 10.5% in group II), with behaviour type A (41% I group and 52% in group II) all presented almost equal values in the two groups regardless of age or BP level. The statistical study of the influence of the risk factors on BP values, using student's t test has demonstrated the contributory part of the genetic factor, the excess of sweets and stress. The deleterious effects of arterial hypertension risk factors, which can begin early in childhood demonstrate the necessity for responsible parties (family, school, doctors, etc.) to be aware of these risk factors and assist in their control. PMID- 12089931 TI - Ender nailing versus external fixation in the stabilization of type III open tibial shaft fractures. AB - Management of severe open tibial shaft fractures presents a difficult challenge to the orthopaedic surgeon. They are frequently associated with loss of limb, infection and high levels of morbidity. All the authors considered, now, that there are five keys to successful treatment: antibiotic therapy, radical debridement and pulsed lavage irrigation, stabilization of fracture with minimal further devascularization, early soft tissue coverage and early bone-grafting. It rests, also, a number of controversies in the management of open tibial fractures, not least of which is the method of fracture stabilization: the choice between intramedullary nailing and external skeletal fixation, the use of reamed or unreamed nails. MATERIAL AND METHODS: Fifty-seven patients with 62 open fractures of the tibial shaft type IIIA, B and C (Gustilo) were treated between 1.01.1994 and 31.12.1998 in the Department of Orthopaedic Surgery of the Emergency Hospital, Iasi, Romania. There were 29 cases type IIIA fractures, 26 cases type IIIB and 7 cases type IIIC (Mess score showed the viability of the limb). Our patients were 36 males and 21 females; their mean age was 36 years (range 17 to 70). Forty-six patients were injured in road traffic accidents and 11 in other traumatic incidents. In 33 cases the skeletal stabilization was achieved by Ender nailing under general or regional anesthesia. In 15 cases we used a bilateral uniplanar external fixator (Burghele) and 14 fractures were stabilized with an Ilizarov external fixator. We note a secondary amputation after the failure of the revascularization of the limb in a type IIIC fracture. RESULTS: All fractures united; the mean time to union was 30.2 weeks--in the external fixation group and 26.4 weeks--in the Ender nailing group. Malunions occurred slightly more frequently in the external fixation group that in the Ender nailing group (15.7% versus 5.8%). We noted--also--more secondary procedures required in the external fixation group. The infection rate was 3 of 33 (9.1%) in the Ender nailing group compared to 4 deep (13.8%) and 8 pin-tract infection (27.6%) in the external fixation group. CONCLUSIONS: Our study suggests that Ender nailing has several advantages over external fixation in the management of severe open tibial shafts fractures. Based on these results, over the last years, in our Department we use mainly the Ender nailing technique, as we consider it a better approach for these type of lesions. PMID- 12089932 TI - [Neurologic manifestation in mumps virus infection]. AB - OBJECTIVE: To study the incidence, clinical manifestations and treatment of mumps virus meningitis. MATERIAL AND METHOD: Study of 64 patients with mumps virus meningitis, admitted to the Clinic of Infectious Diseases of Iasi during 1996 1999. RESULTS: The mumps virus meningitis had a high prevalence in 1998 (39%) most patients being males (62.5%) from the urban areas. 39.06% of cases were on the age group of 5-6 years old, 25% were in the age group 10-14 years old and 18.75% were adults patients. The clinical manifestation were dominated by headache (100%), fever (100%), vomiting (89%), photophobia (43.75%) and meningeal syndrome of contracture. The laboratory diagnosis was established by the cerebro spinal fluid exam, together with the peripheric blood exam. The treatment was in all the cases with pathogenic (corticotherapy) and symptomatics with a very good evolution. CONCLUSIONS: All the cases with diagnosis mumps virus meningitis were well going, having good evolution and complete healing under symptomatic and pathogenic therapy. PMID- 12089933 TI - [The luteolytic action of prostaglandin F2-alpha analogues in guinea pig's ovary]. AB - The role of prostaglandin at the level of the female genital system is multiple; they are involved in ovulation, luteolysis, contractility of the tube muscles, contraction of the gravid uterus, dilatation of the cervix. The synthetic analogous of the prostaglandin F2 alpha are utilized in a large area of treatments, for multiple purposes. The study was made on 2 lots with 10 non pregnant female guinea pig: the first lot was the witness lot and didn't receive any substance. The second lot received 100 micrograms/kg/day Isopropyl ester PGF2 alpha. The prostaglandin analogous produces vasodilating effects on the ovary blood vessels and luteolytic effects with the destruction of the ovarian follicles. PMID- 12089934 TI - [Ultrastructural characteristics of the adrenal gland]. AB - The Adrenal glands can be viewed as sustaining vital processes in the human body, through two different but related components: the cortex and the medulla. It is well known that the close connection between the cortical cells is able not only to support the adrenal functions, but in the same time to serve the better understanding of the normal way of its hormonal production. Using the ultrastructural study of the adrenal gland, we obtain images which can provide informations about details of the cellular structure, in the eventuality that all this work will continue to improve to the point that we might be able to completely understand the complex interrelations between the structure of each cell and the function of the whole organ. PMID- 12089935 TI - [Pre-test and post-test evaluation of HIV/AIDS educational program for adolescent]. AB - The increasing incidence of AIDS among young people involves the need for effective HIV/AIDS prevention programs. The evaluation of such a program, using pretest, information and posttest, was performed in a high-school of Iasi City, on a sample of 169 pupils. They answered a semi-structured questionnaire, concerning false and real HIV ways of transmission, attitudes and pre-conceptions about HIV infected people and preventive knowledge. The findings of this study showed that the cognitive level increases with age (higher at 17-19 years old pupils) and the receptivity is higher at 15-16 years old pupils. PMID- 12089936 TI - [The relationship between benign breast proliferation (fibrocystic mastopathy) and the appearance of breast carcinoma]. AB - Fibrocystic mastopathy (FM), a benign lesion, is considered by many author like a premalignant lesion. Breast carcinoma (CA) represents the most frequent female tumor and the second mortality cause through cancer, after the bronchopulmonary cancer. At the Department of Pathology from Tg. Mures, we selected 190 cases, for 10 month: 102 presented FM, 41 CA and 47 CA + FM. In every case, we have examined 2 slides stained with HE. In difficult cases, we performed special stains (van Gieson, H-PAS, Alcian). We studied the pulmonary tumor, adjacent mammary tissue and axillary lymph nodes. The present data of literature and the experience of the Department of Pathology from Tg. Mures showed a strong relationship between the presence of FM and the appearance of CA: both lesions have a common field of hormonal disfunction and intraductal and intralobular hyperplasia (an microscopic feature of mastopathy) increase the risk of CA. PMID- 12089937 TI - [Genetics of the polycystic ovarian syndrome and therapeutic perspectives ]. AB - PCOS is the most frequent endocrine disorder of premenopausal women. The common clinical signs of PCOS are hirsutism, menstrual irregularities with chronic anovulation and a trend toward overweight or obesity. Diagnosis is based upon high plasma levels of androgens and the ultrasound image of polycystic ovaries. The high prevalence of PCOS at first degree female relatives suggest an important genetic component of this syndrome. Linking studies in sisters presenting phenotypical traits of PCOS and in their parents allowed the investigation of certain candidate genes presumed to be involved in the physiopathology of PCOS. The genes encoding enzymes involved in androgen synthesis, protein transducers of insulin signals and the paracrine regulating factors of gonadotrophins and ovarian function have been analysed. To date, no determinant gene mutation was reported. However, several loci were detected, especially a locus within the insulin receptor. Mutations or gene polymorphisms and their function remain to be identified. These research attempts should explain the physiopathology of PCOS and open new therapeutic perspectives. The usage of medication increasing the sensitivity to insulin action is an example of applying these particular aspects. PMID- 12089938 TI - [Morphological study of osteosarcomas in children]. AB - Osteosarcoma is the most frequent primary osteal tumor in children. Our study evaluates morphological aspects of osteosarcomas during last 20 years. We present the results of study that includes 32 cases of classical osteosarcomas that had been diagnosed and treated at the "Sf. Maria" Hospital from Iasi (Romania). The diagnosis is based on usually histopathological methods applied to bioptic and surgical exeresis pieces. Male sex is most involved (56%). Patients' age is between 12-15 years in 68% of total cases. In 50% cases the tumour was localized in distal femoral metaphysis. Histological forms were: osteoblastic (66%), chondroblastic (18%), fibroblastic (16%). Enneking's staging was: II A (6.25%), II B (81.25%), and III (12.5%). Metastasis appeared in 75% cases' evolution. Concluding, 25% of studied patients had a favourable evolution during a 5 years period. Survival rate at 2 years was 39%. PMID- 12089939 TI - [Bactericidal activity of Trovafloxacin and Moxifloxacin against Gram-negative anaerobic bacilli]. AB - Anaerobes are often involved in human pathology, including oral infections. The therapy of anaerobic infection requires certain antibiotics such as: metronidazole, tetracycline or amoxicillin. But the incidence of resistant strains among anaerobes is continuously increasing and it is necessary to find some new agents effective against these bacteria. The aim of our study was to establish the bactericidal activity of two new quinolones against 14 strains of gram-negative anaerobic bacilli. The MIC of tested strains was first detected and then time-kill kinetics was determined by using Miles and Misra methodology. Ciprofloxacin was used as standard-quinolone. The results show that trovafloxacin and moxifloxacin have good activity against anaerobes, better than that of ciprofloxacin. Bactericidal curves suggest a rapid action of moxifloxacin (within the first 4 hours), but a prolonged cidal activity for trovafloxacin (24 hours). For both agents, the antimicrobial effect is concentration-dependent. As previous studies suggest, the new quinolones have good activity against both aerobic and anaerobic bacteria so they are useful in the treatment of mixed infections (common feature of anaerobic infections). PMID- 12089940 TI - [National statistical parameters of cervical cancer in Romania: 1992-1997]. AB - Cervical cancer is an important health issue in Romania, as the 1990-1992 statistics of IARC mentioned it to be first in Europe as mortality through this neoplasia. We bring to 1997 the national statistical data regarding the cervical cancer. MATERIAL AND METHODS: Our data provided by the Romanian Health Ministry statistical Office allowed estimations over the following parameters: incidence (crude and age-related), prevalence and number of deaths, and relationships between these parameters and age group, clinical stage, geographical distribution, etc. Usual statistical methods of interpretation are used, as well as prognosis using the parabola methods, and world-standardized incidence rates. RESULTS: Between 1992-1997, there were 14743 new cases of cervical cancer diagnosed in Romania. The crude incidence per 100,000 was 21.27, and age-related incidence ranged between 0.27 per 100,000 for the under 20 to 46.73 in the 40-49 group. World standardized incidence rate indexed by age groups gave an incidence of 16.84. The mortality was 2.37, with world standardized age related deaths of 2.81. The clinical stage distribution showed stages 0 (15.02%), I (14.04%), II (34.91%), III (27.81%) and IV (8.19. The incidence and mortality data shows a better evolution for younger age groups, but a steady increase in 30-39 group for incidence and mortality. CONCLUSIONS: The data above place Romania on a stable but high incidence of cervical cancer, with a high mortality due to late diagnostic and socio-economical conditions. PMID- 12089941 TI - [Researches on pharmacological properties of some new xanthine derivatives]. AB - We studied the potentially bronhodilatatory action of some new xanthine compounds, theophylline derivatives with 7-[2-hydroxy-3-(4-acetamido)-phenoxy propyl]-8-R-1,3-dimethyl-xanthine and 8-R-1,3-bis-(1,3-dimethyl-xanthin-7yl)-2 hydroxy-propane structure substituted in 7 and 8 position. The tracheal smooth muscle contraction we realized by carbachol (10(-5) M) or potassium chloride (40 mM), the xanthine compound being much active on the contraction induced by carbachol. We observed that the modification realised in theophylline structure determined the modification of bronchodilatatory properties. The bronchodilatatory effect of theophylline intensified with the introduction in 7 position the 2-hydroxy-3-(acetamido)-phenoxy-propyl and in 8 position the bromo, nitro, piperidinyl and imidazolyl radicals. PMID- 12089942 TI - [The influence of some synthetic prostaglandin F2alpha analogous on rat lung]. AB - The role of prostaglandins and their synthetic analogous at the level of the female genital system is multiple. The synthetic analogous of the prostaglandin F2 alpha are utilised in a large area of treatments, for multiple purposes, but their influence on the lung is not so well known. The study was made on 2 lots with 12 non pregnant female rats: the first lot was the witness lot and didn't receive any substance. The second lot received 25 micrograms/kg/day Flavoliz (synthetic analogous of the prostaglandin F2 alpha). The prostaglandinic analogous produces vasodilating effects on the lung blood vessels and moderate pulmonary destruction. PMID- 12089943 TI - [Advances and limits in biomedical stress quantification in dento-periodontal structures]. AB - The stress quantification in dento-periodontal structures is an important concept, as stress in the tissue and in surrounding structures. A number of clinical studies have suggested figures for such an optimal stress strange. The clinical experiments can be completed with mathematical analysing methods as finite element method. Once the basics of biomechanics understood laws the treatment goes clearly defined, the computer aided design improve the optimal restorative prosthetic or orthodontic appliance. The finite element method (FEM) makes possible the numerical simulation of different clinical situations. Based upon mathematical modelling and extrapolation from experimental studies, practical conclusions can be drawn concerning the different types of forces and their subsequent induced relationship to the stresses. Clinical and histological experiments confirm the mathematical prediction offered by the FEM. PMID- 12089944 TI - [Periodontal regeneration with enamel derivate proteins (Emdogain)--clinical evaluation]. AB - The study was undertaken in order to evaluate the effect of Emdogain as an adjuvant in periodontal surgery in promoting bone level gain, compared to bone levels after surgery. METHOD: The study was realised on 6 patients. who had pair test sites and control sites. Two interproximal sites were selected in each patient at a distance from each other, in the same maxilla, with infra-osseous defects with depth > or = 4 mm and width > or = 2 mm. The patients were followed on a long term period, initially and after 8 and 16 months. RESULTS: The average values of the clinical attachment level were evaluated both in the test sites and in the control group at 8 months and 16 months, The radiographic osseous level continued to recede at the group treated with Emdogain, and in the control group it remained at its initial level. The radiographic gain was ascertained after 16 months. CONCLUSION: This study proves that the topic use of Emdogain as an adjuvant, during periodontal defects surgery, on radicular surfaces, determines a gain in the clinical attachment and an osseous gain observed on X-rays. PMID- 12089945 TI - [Difficulties in the diagnosis of prolonged fever in infants]. AB - We report the case of an infant with cytomegalovirus (CMV) infection who developed a nosocomial sepsis of mixed etiology. Fever failed to decrease despite long-time and sustained antibiotic therapy. Treatment with an antiviral agent following the detection of CMV antibody of the IgM class resulted in clinical improvement and subsequent full recovery. We suggest that the diagnosis of CMV infection should be considered even in immunocompetent patients in whom antibiotic therapy fails. PMID- 12089946 TI - [An unusual case of multiple endocrine neoplasia]. AB - The patient A.M., woman, presented Multiple Endocrine Neoplasia, associating hyperparathyroidism, an adrenal tumor and pancreatic tumor; one can observe that the disease had features of Multiple Endocrine Neoplasia type-1 (pancreatic tumor), as well as Multiple Endocrine Neoplasia type-2 (MEN-2, pheochromocytoma), while the hyperparathyroidism is common for both two types of the syndrome. The development of the disease is extended over approximate forty years of life and finally led to death. The first symptoms appeared during pregnancy, at age 26, when the severe hyperparathyroidism determined bone mass acute loose, with decrease in height by affecting columnary vertebrae, severe bone pain due to bone resorption and soon after symptoms of renal stone disease. The patient suffered two surgical interventions for renal lithiasis. The biliary symptoms required also the surgical removal of the gall bladder, putting the diagnosis of active metabolic stone disease. At age 56, was established the diagnosis of carcinoma of the adrenal cortex, which was solved with surgical therapy. At age 66, digestive symptoms suggested the presence of a carcinoid tumor. Abdominal ultrasonography revealed a well-lined pancreatic tumor of 2-3 centimeters in diameter, probably adenoma. The malignant evolution of this tumor was rapid, leading to death in less than two years. The constant refuse of other investigations, by the patient, made us incapable to determine the type of secretion of the pancreatic tumor; we rather suspected a VIPoma, because of the watery diarrhea and severe weight loss. The most difficult to treat was anyway the hyperparathyroidism and its complications. PMID- 12089947 TI - Gas chromatographic detection of Mycobacterium tuberculosis complex in pure cultures from respiratory specimens. AB - It is important to identify mycobacteria to the species level in order to establish their clinical significance and to take the appropriate therapeutical decision. Biochemical tests on primary cultures take time (3-6 weeks) until the report of results; that's why more rapid techniques are needed. We have used gas chromatography with flame-ionisation detection (GC-FID) as an alternative identification method for 53 mycobacterial strains isolated from respiratory specimens. We have extracted fatty acids from whole mycobacterial cells, then derivatized them into methylesters, detectable by GC-FID. All the strains were identified as M. tuberculosis complex (MTC), using the Microbial Identification System (MIS) software. The specificity of the identification by GC-FID of MTC is 100%. In conclusion, pulmonary mycobacteriosis are dominated by MTC; GC-FID is a rapid and accurate method for the identification of MTC. PMID- 12089948 TI - Histology teachware for the Web site of the University of Medicine and Pharmacy "Gr. T. Popa" Iasi. AB - Design and implementation of Web educational materials for medical sciences are discussed starting from a brief presentation of the current trends in using Internet facilities and placing emphasis on the specific case of the www page constructed for the Department of Histology. The paper focuses on the development of teaching-aids for the morphological study of organs and systems, illustrated by the creation of Histology Album, including photographs and explanatory texts, which is accessible via this www departmental page. Relevant aspects for the construction of the Histology Album are detailed in terms of both logical structure and physical implementation, based on HTML techniques. Outcomes are expected in achieving a noticeable progress in students' instruction, as a result of diversifying the documentation possibilities and increasing the role of the individual work (by avoiding the permanent supervision of an instructor). Further improvements of this www page are commented in the light of the recently proposed frameworks for distance learning, relying on specialized software tools that allow student's interactive work. PMID- 12089949 TI - [The 7th World Conference of Clinical Pharmacology and Pharmacotherapy]. PMID- 12089950 TI - [The 9th International Congress on Magnesium MAG 2000]. PMID- 12089951 TI - [Psychiatry 2000. Traditions and perspectives. The National Congress of Psychiatry]. PMID- 12089952 TI - [An almost forgotten personality of the medical education of Iasi - professor doctor Petrus Niculescu]. PMID- 12089953 TI - [How I became pediatrician]. PMID- 12089954 TI - [Renal osteodystrophy(II)]. AB - Renal bone disease represents one of the major complications of end-stage renal disease, accounting for the numerous and various changes at bone level, determined by abnormal calcium and phosphorus homeostasis and by changes in calcitriol and PTH synthesis. PTH represents as well a major uraemic toxin, exerting profound systemic effects, particularly at the cardiovascular level. PTH synthesis is mainly controlled by changes in calcium-phosphorus balance and calcitriol production by the kidneys. Several others factors are important in the development of secondary hyperparathyroidism: acidosis, autonomisation of PTH secretion and peripheral (target-organ) resistance to PTH actions. Although bone biopsy represents the definitive diagnostic test to differentiate between osteitis fibrosa, low-turnover bone disease and bone involvement unrelated to disturbed calcium metabolism (i.e. beta 2-microglobulin-related amyloidosis), plasma intact PTH generally exhibits a reasonably good relation with bone histology parameters. Moreover serum bone-specific alkaline phosphatase isoenzyme, serum pyridinoline and the novel serum markers for bone turnover are highly specific and correlate with bone histomorphometry parameters, so that, preventive and therapeutic strategies should be re-evaluated based solely on biochemical parameters. PMID- 12089955 TI - [Radiobiological considerations in radio-induced soft tissue sarcomas]. AB - Soft tissue sarcomas are relative rare tumors with an incidence of almost 1% of all malignancies. Although their etiology is not well understood, studies regarding the effect of ionizing radiation, showed their capacity to induce sarcomas in soft tissues, after several years from the treatment of other primary tumor. Their early diagnosis could be achieved by genetic analysis of radio induced chromosomal translocations specific to each histological type. Dose response curves reveals the dynamics of the process, taking into consideration the beam energy and linear energy transfer. PMID- 12089956 TI - [Preoperative and intraoperative scintigraphic detection of sentinel node]. AB - It is known that the malignant primary tumor site has a specific lymphatic drainage, all the time intermediate, however, by a first lymph node named sentinel node. This can be situated in some typical or atypical topographic area and can be or not clinically evident. If the sentinel node is not invaded by the metastatic cells there is no metastatic risk for other lymphatic nodes, so the surgical treatment is not necessary to involve the corresponding lymphatic area dissection. An imagine, noninvasive detection of the involvement of the sentinel node in the spread of cancer could be related with the surgical decision to perform (or not) fully regional lymphadenectomy (based on biopsy result of such positive detected nodes) allowing only a tumorectomy in the first stages of neoplasia. For the case of scintigraphic method to detect the sentinel node, the performance depend on the development of radiotracers, the modification of the lymphoscintigraphic mapping techniques protocols, and the development and improvement of detectors. PMID- 12089957 TI - Implications of imidazolines and imidazoline receptors role at the vascular level. AB - There are both post- and pre-synaptic vascular imidazoline (IM) binding sites. The importance of direct IM actions and that of peripheral imidazoline receptors (IRs) are shadowed by the central effects of IMs and by their interaction with alpha 2 adrenoceptors. Since the discovery of clonidine the many studies on IMs have been focused on their hypotensive effect, with rilmenidine and moxonidine as representative drugs. Formerly called IM preferring alpha 2 or IM/guanidium sites, the IRs (idazoxan-sensitive) are the plasmalemmal I1 (clonidine-sensitive) and the various I2 (one structure identified as MAO). I1 signaling includes activation of phosphatidylcholine-selective phospholipase C and inhibition of some ligand-gated channels. Inhibitory IRs on postganglionic sympathetic terminals, are not alpha 2, H3, I1 or I2. Some IMs directly affect CaL, while others inhibit K+ efflux. Clonidine-displacing substances including agmatine are endogenous ligands at IRs and alpha 2 and may participate in arterial pressure control. Beside few speculations, the roles of vascular IRs are largely unknown. PMID- 12089958 TI - New derivatives of m-nitrobenzoyl-asparagic acid and m-nitrobenzoyl-asparagine with anti-inflammatory action. AB - In order to obtain new biologically active compounds some derivatives of 1-[N-(m nitrobenzoyl)-alpha-D,L-asparaginyl]-2-benzyl-benzimidazole have been synthesized by ring opening reactions of 2-(m-nitrophenyl)-4-(beta-amidomethyl)-delta 2 oxazolin-5-one and 2-(m-nitrophenyl)-4-(beta-carboxymethyl)-delta 2-oxazolin-5 one with 2-benzylbenzimidazole (dibazole). PMID- 12089959 TI - [The examination of the digestive tract in patients with iron-deficiency anemia]. AB - According to literature the gastroenterologic consultations for iron-deficiency anemia are quite frequent. The aim was the evaluation of the part played by gastrointestinal examinations for the diagnosis of iron-deficiency anemia. There were 115 patients admitted in the Medical Clinic between 1998-1999, with iron deficiency anemia in the absence of macroscopic bleeding who carried out upper or lower endoscopy. A digestive lesion which account for iron deficiency anemia was identified in 35% of the cases. In the upper digestive tract there were 60%, in decreasing order of frequency: peptic ulcer, gastric cancer, erosive gastritis, angiodysplasia. In the lower digestive tract were 35% of the cases (colorectal cancer, polyps, angiodysplasia and hemorrhoids). In 5% of the cases there were found synchronization of the digestive tract lesions. So the digestive tract examination is worth doing because it establishes the diagnosis in 35% of the cases with few symptoms. The digestive tract lesions are more frequent in the upper tract, but they are more severe in the lower digestive tract. The possibility of the synchronization of lesions proves the necessity of carrying out the complete examination of the whole digestive tract. PMID- 12089960 TI - [Clinical characteristics of patients with ischemic heart failure]. AB - A major cause of heart failure is coronary heart disease with an increasing prevalence and progression to heart failure. The objective of the study is the analysis of the clinical peculiarities, coronary risk factors and some resting hemodynamic parameters at patients with ischemic heart failure. METHODS: There have been studied 189 patients with ischemic heart disease. The ejection fraction (EF) assessed by echocardiographic examination was used to divide them into three groups: group A of 106 with EF above 50%, group B of 55 with EF between 40-49% and group C of 28 with EF below 40%. RESULTS: The mean age increased from group A (58 +/- 13 years) to group C (64 +/- 11 years, p = 0.05). Ischemic cardiomyopathy was higher at group C (32% vs 13% at group A and 24% at group B, p = 0.01) and old myocardial infarction was more frequent at group A (44% vs 45% at group B and 36% at group C, p = 0.009). Group C revealed a greater incidence of dyspnea (100%) and a greater resting heart rate (83 +/- 12/min, p = 0.03). The radiologic cardiomegaly was present at group C and at 42% at group A (p = 0.001). The echocardiographic findings show a greater prevalence of kinetic abnormalities and diastolic dysfunction at group C (p = 0.002 respectively 0.009). CONCLUSION: Patients with myocardial dysfunction are older, with a lower prevalence of myocardial infarction in contrast with a higher prevalence of cardiomyopathy. Dyspnea is the dominant symptom and the resting heart rate is higher. Systolic dysfunction is present at all patients and diastolic dysfunction at the majority. There were not differences concerning the prevalence of the coronary risk factors. PMID- 12089961 TI - The value of echocardiography for the assessment of the cardiac impairment in chronic renal failure in child. AB - Chronic renal failure (CRF) might gradually induce the occurrence of morphological and functional cardiac disturbances; these cardiac disturbances make up supplementary risk factors in children with CRF. The aim of our study was to determine the role of echocardiography(Echo) for the diagnosis of these cardiac disturbances acquired during the evolution of CRF in child. We studied 20 patients(aged between 5 and 18 years) with CRF, from whom 13 were dialysed. All patients underwent 2D echocardiography with Doppler, including the study of systolic and diastolic function of the left ventricle(LV). The main echocardiographic findings were:--concentric LV-hypertrophy(8 cases) and asymmetrical LV-hypertrophy(3);--dilated cardio-myopathy(3);--diastolic dysfunction of the LV(10);--normal systolic function in most of the cases;- mitral and tricuspid regurgitation(3);--pulmonary hypertension(2);--pericardial effusion (8) to cardiac tamponade (2). Echo allowed a better assessment of cardiac morphology and LV function and their improvement in some cases, after hemodialysis and antihypertensive therapy. CONCLUSION: Echo exam has a great utility for the diagnosis of cardiac disturbances in children with CRF, especially in cases with severe cardiac involvement and without cardiac symptoms; this investigation has to be performed right since the initial diagnosis, as well as for long-term surveillance. PMID- 12089963 TI - [Pharmacogenomics - a promise in clinical practice]. PMID- 12089962 TI - [Osteoarticular changes in childhood leukemia, lymphoma and neuroblastoma]. AB - Osteoarticular changes may occur in up to 23% of the cases with Acute Lymphoblastic Leukemia (ALL) and even more frequent with Acute Myeloblastic Leukemia (AML). Most of the bone and joint pains are due to neoplastic infiltration, radiologically obvious as metaphyseal clear stripes, parietal enlargement, periosteal reaction, osteolysis and diffuse osteoporosis in the long bone. In Malignant Lymphomas (ML) the bone involvement is rarer, usually bone metastases identifiable with Tc scintigraphy being the cause. In Neuroblastoma (Nbl) cases, bone metastases are commonly associated with abdominal tumor beyond one year of age. Of the total ALL, AML, ML and Nbl cases treated in our Oncology Dept, we selected 43 children with osteoarticular involvement. The sex ratio was 24 boys to 19 girls and the specific malignancy was ALL in 25 cases, AML in 5 cases, ML in 2 cases and Nbl in 10 cases. The following biological parameters were monitored: type of onset, the localization and nature of the bone affectation, differential diagnosis, and the response to therapy. The presence of the osteoarticular involvement has proved to be of no prognostic significance. The spectrum of clinical manifestations varied from mild pain to severe disability, in 7% of the cases being the unique symptoms. The type of lesion did not rise important differential diagnosis issues, excepting the cases with unique osteolytic lesion or diffuse osteoporosis, where the rest of the data and the elevated urine vanilmandelic acid helped to establishing the diagnosis. We conclude that the osteoarticular involvement encountered in different malignancies in children is a major sources of diagnostic problems, but it is not associated with a significant outcome. PMID- 12089964 TI - [EEG patterns in alcohol withdrawal syndrome]. AB - The cerebral motor and cognitive behavior is obviously altered by chronic alcoholism, which generates significant morphological and biochemical changes. In turn, alcoholic withdrawal improves the cerebral blood supply, the psychological and motor behavior and the EEG appearance, especially in young alcoholics. Our study also underlines that alcoholic women develop a late improvement of the clinical and bioelectrical condition during the alcoholic withdrawal. PMID- 12089965 TI - [The evolution of the natural-killer-dependent cytotoxic activity during normal pregnancy]. AB - The fetus was compared to an allograft because paternal antigens are also expressed. Even the analogy is not a real one, the main steps of the rejection phenomenon are relevant for the understanding of the complex materno-fetal interactions as the large granular lymphocytes are the first immune cells which invade the graft. Due to the important role of NK cells in the cell mediated immune response, the purpose of this paper was to assess the NK cell dependent cytotoxicity during normal pregnancy. The study was performed on a test group of 52 normal gravidas, in the Ist, IInd and IIIrd trimester and a control group of 20 healthy nonpregnant women with similar age and parity averages. The cytotoxic activity of the NK cells was measured in a 51Cr release assay against K 562 cell line. The results were expressed as the cytotoxicity index (%) and showed a statistically significant decrease for all gravidas compared to controls, mostly for the IInd trimester. The trend of the NK cytotoxic activity during normal pregnancy suggests the deep involvement of the NK cells in the mechanisms acting at the fetomaternal interface. PMID- 12089966 TI - [Chronic scapulo-humeral instability in children]. AB - The recurrent dislocation of the shoulder is a rare condition during childhood, and it should benefit from an early treatment in order to prevent secondary lesions of the humeral head from appearing. Osteoarthritis in adulthood is a potential complication of this disease. A number of 10 children (5 boys and 5 girls) were treated in our department between 1989 and 1998; 8 of them had the anterior type of dislocation, while 2 children had the posterior type. All children were older than 10. We performed the anterior Bankart procedure in 6 children, the posterior Bankart procedure in 3 cases and Putti-Platt procedure in 2 children, respectively. Surgery was followed by casting for 3 weeks and physical therapy afterwards. We obtained very good results in 4/6 cases. Recurrence appeared in 2 cases--one of them failed to respond again to surgery, the other one refused reintervention. We conclude that a simple and well performed surgical procedure is recommended in children, followed by physical therapy. Psychotherapy can be required if a voluntary component is implicated in producing the dislocation. PMID- 12089967 TI - [Incisional hernias after operative laparoscopy]. AB - The objective of this study was to determine the possible risk factors of incisional hernias after operative laparoscopy. Complications involving the abdominal wall, particularly incisional hernias, were not expected when laparoscopic procedures were first introduced. With the increasing number of laparoscopies in the abdominal surgery, more incisional hernias were observed. Of the approximately 3,145 patients who underwent laparoscopy from march 1993 through march 2000 in the First Surgical Clinic Iassy, a total of 15 hernias occurred, which is an incidence of 0.477%. There were no acute complications, such as small bowel obstruction, secondary to herniation through the trocars. Risk factors such as chronic bronchitis or weight increase, which give rise to abdominal pressure, were present in some cases. Precautions such as fascial suturing must be taken to reduce the incidence of this minor complication of laparoscopic surgery. PMID- 12089968 TI - [Immediate and late complications of transurethral resection of prostate]. AB - 300 patients with benign prostatic hyperplasia admitted in hospital between 1994 and 1998 were studied. The surgical procedure in 21 patients was TUIP (transurethral incision of prostate), in 18 patients was classic adenomectomy (open prostatectomy), in 58 patients classic TURP (transurethral resection of prostate) and in 203 patients TURP by low pressure (hypopressure). The immediate complications in those patients who underwent endoscopic procedures were bleeding in 18 patients, acute urinary infections in 20 patients, endotoxic shock in 5 patients, epididimytis in 9 patients, urine retention in 4 patients, obstruction of catheters in 10 patients, collection of washing liquid in perivesical space (Retzius) in 3 patients. The late complications were the following: bleeding--4 cases, imperative micturition--12 cases, vesicle neck sclerosis--3 cases, urethral orifice stenosis--16 cases, urethral stricture--3 cases, urinary retention--6 cases, abscess in perivesical space (Retzius)--1 case. After TURP in hypopressure, the complications were less frequent due to a better visualization a more rapid execution of the procedure. PMID- 12089969 TI - [Laparoscopic cholecystectomy in non-lithiasis cholecystopathies]. AB - We made a study in 1st Surgery Clinic of Iassy between March 93--November 98, with 2246 laparoscopic cholecystectomies. The reason was to define the place and role of laparoscopy in non-lithiasis cholecystopathies (81 cases). In the same period were operated classically 888 calculous cholecystitis and 38 non-lithiasis cholecystitis. The laparoscopy has a major role in diagnosis to the patients with many abdominal symptoms. We discovered pericholecystic adherences, hepatic cirrhosis, acute appendicitis, etc. Sometimes, the laparoscopy was made for "second look" after surgical treatment for neoplasia, the metastasis diagnosis, for tumors visible echographically. In many situations the laparoscopic cholecystectomy may be considered like a preventive operation. The easy postoperative evolution is an argument to enlarge the indication for laparoscopic cholecystectomy in alithiasis cholecystitis. PMID- 12089970 TI - [Marjolin's ulcer on burn scar, a curable but neglected disease]. AB - We discuss 9 consecutive carcinomas developed on postburn scars. Our interest was focused on surgery, recurrence, metastasis and long-term survival. The delay between burn trauma and the first clinical manifestation was 25-63 years. The most common localisation was in the limbs (10). Two cases presented with visceral metastasis. We performed either wide excision and grafting (5) or amputation (5). From the anatomo-pathological stand point we encountered 8 SCC and 1 BCC. In 6 cases we had no local recidive or metastasis. One presented a local recidive. There were two deaths--lung metastasis and "spontaneous" rupture of invaded axillary artery. The Marjolin's ulcer has a low incidence because of the patient's poor education. The prophylactic attitude is optimal. We favour early excision--grafting of the deep burns, long-term follow-up, excision and grafting of unstable areas (joint area, depigmented regions, chronic ulcers). The optimal surgical technique is excision followed by skin grafting since it allows early detection of the recurrence. PMID- 12089971 TI - [Multimodality treatment outcome of invasive thymomas]. AB - The aim of the study was to analyze the progression of invasive thymomas associated with myasthenia gravis, after the resection and the progression of unresectable invasive thymomas with a combined chemoradiotherapy. The study was performed on two groups of patients: 8 patients with invasive thymomas and myasthenia gravis operated at the 3rd Surgical Clinic between 1986-1999; 4 patients with unresectable invasive thymomas treated at the Radiology-Oncology Clinic by combined chemoradiotherapy, between 1993-1998. The results are presented for each group of patients, separately. CONCLUSION: The best treatment of invasive thymomas is the multimodal one. The timing of each method was established based on the collaboration between surgeons, medical oncologists, radiotherapists and neurologists, depending on the characteristics of each patient. PMID- 12089972 TI - Resistance mechanism for a number of non-typhoidic Salmonella strains isolated in Iasi, Romania. AB - The aim of this study was to find a possible explanation for the resistance pattern of the selected strains and to evaluate the impact in case of isolating such strains. During the period February-March 1998, in the Pediatric Hospital "Sf. Maria" from Iasi, Romania, 12 non-typhoidic Salmonella strains, non duplicates were isolated from stools of newborns and, in one case, from mother as well. Identification was done by standard methods, and sensitivity testing by disk diffusion method, against the following antimicrobials: Ampicillin (A), Amoxycilin/Clavulanic Acid (AMC), Tetracycline (T), Streptomycin (S), Cloramphenicol (C), Nalidixic Acid (NA), Ciprofloxacin (CIP), Cephalotin (KF), Cefuroxime (CXM), Ceftriaxone (CRO), Cefotaxime (CTX), Ceftazidime (CAZ), Aztreonam (AZT), Imipenem (IPM), Amikacin (AK). All strains showed resistance to: A, AMC, T, S, C, NA, CRO, CAZ, CXM, CTX, KF except one strain, CTX and CRO sensitive. The possible explanation is the production of an extended-spectrum beta-lactamase (ESbLA), detected by microbiological methods (Masuda and clover leaf). Resistance transfer was realised by using the previously described method. In conjugation experiments we have obtained only one transconjugant, identified as E. coli, that has acquired all resistance determinants. This report highlights on the isolation of multiresistant non-typhoidic Salmonella strains in hospitalised patients without previous antibiotic treatment and possibility of detecting such strains by being aware of the correlation: resistance phenotype resistance mechanism. PMID- 12089973 TI - [Contribution of molecular biology on the progress of pharmacology and therapeutics]. PMID- 12089974 TI - [Occupational stress--risk factor in essential arterial hypertension]. AB - The stress was recorded taking into consideration the epidemiological interview, using a standard methodology by filling in the type card the occupational stress (OS) and the family stress (FS). The constitutional factors (behaviour type A, body weight index > 25 Kg/m2) as well as the consumption in excess of salt, have significantly correlated themselves, both in individuals with OS and FS, and in those with the exclusively OS. The modification of biochemical constants (total cholesterol, triglycerides, total lipids, HDL-cholesterol, LDL-cholesterol) significantly associate themselves in persons to which the presence of OS and FS, of the occupational one, respectively was put into evidence (chi 2 = 94.10 for 5 freedom degrees and p < 0.001, chi 2 = 118.94 and p < 0.001, respectively). The prevalence of persons with pathologic values of total lipids was 58.7%, of triglycerides 35.7%, of total cholesterol 12.7%, of LDL cholesterol 27%, and of HDL-cholesterol 44.4%. Measuring the values of systolic and diastolic arterial hypertension an increase above the normal values of limit systolic arterial hypertension at 23.8% and at 12.3% for illness systolic arterial hypertension was found. The evaluation of association of certain cardiovascular risking factors in individuals with occupational stress has permitted a global understanding, useful for the elaboration of some preventive programmes. PMID- 12089975 TI - [Influence of corrosion products of the dental alloys on the excitable tissues]. AB - The surface of dental alloys in contact with well conducting electricity solutions are electrochemically corroded. This phenomenon is determined by a proper electric wave generated by electrochemical phenomena present at the limit between the two phases. Because during dental treatments similar conditions with the generation of corrosion waves are present, we decided to study the electrochemical behaviour of the most commonly used dental alloys. The method used in our study is based on the anodic and catodic reactions developed simultaneously, represented by the global curve of polarization that gives informations on the intensity of corrosion waves. We have also studied the influence of corrosion products on the excitable tissues. The results demonstrate the significant influence of metallic ions on the excitable tissues. PMID- 12089976 TI - [Comparison on the effects of metronidazole in subgingival application in the treatment of adult periodontitis]. AB - Control of subgingival plaque is the key of the treatment of periodontal diseases. The use of subgingival sustained-release antibiotic therapy is advocated for the beneficial effect on anaerobic flora. A single-blind clinical trial was carried out in 13 patients with adult periodontitis to compare the effects of subgingival application of metronidazole dental gel with those of subgingival scaling. METHOD AND MATERIALS: A split mouth design was used so that each patient received all treatments simultaneously. Randomly selected quadrants were treated with applications of 25% metronidazole gel, subgingival scaling, or a combination of scaling and gel application. The remaining quadrant in each patient was left untreated as a control. RESULTS: All three treatments were effective in significantly reducing bleeding on probing over the 14 weeks observation period. No statistically significant differences were found between scaling alone and combined treatment. Scaling and combined treatment were better than metronidazole. Metronidazole produced transient effects, best noted during the first 4 weeks after treatment. No additive effect of metronidazole was noted in the combined treatment. At week 14, only combined treatment sites and scaled showed statistically greater probing depths reduction than control sites. CONCLUSION: For the treatment of mild to moderate adult periodontitis, subgingival scaling alone is as effective as the combination of scaling and antibiotic therapy. PMID- 12089977 TI - Sensitivity of Enterobacteriaceae strains to the third generation cephalosporins. AB - The aim of this study was to evaluate the in vitro activity of cefotaxime, ceftriaxone, ceftazidime, ceftibuten and aztreonam against the clinical isolates of Enterobacteriaceae. MICROORGANISMS: A total of 973 enterobacteria isolated from clinical material (urine and faeces) were tested. Strains were collected during 1996-1998 period, in some centers of Eastern Romania. SENSITIVITY STUDY: MICs were performed using the agar dilution method following standardized techniques (NCCLS, 1994). Stock solutions of antibacterial agents were prepared from their respective powder forms, in concentrations ranging from 0.03 to 128 mg/ml. Our results show a good activity of the tested antimicrobials on Escherichia coli, Shigella spp and Klebsiella spp tested, and reveal a particular behaviour for Salmonella spp, characterized by a heterogenous susceptibility. In conclusion, cefotaxime, ceftriaxone, ceftazidime, ceftibuten and aztreonam were active at low concentrations against tested isolates. Against Salmonella spp, these agents were less active: percent of resistant strains by beta-lactamase production was significant. PMID- 12089978 TI - [Seizures during the treatment with interferon for chronic C hepatitis]. AB - Interferon alpha (IFN-alpha) is a well-established first-line treatment for chronic viral hepatitis. Side effects of IFN-alpha therapy are common but generally mild and self-limited. Generalized seizures during IFN-alpha therapy are very uncommon and are present in clinical isolated cases and usually in association with high doses of IFN-alpha. In our case a female of 39 years old, seizures have occurred at low doses of IFN-alpha used as therapy for chronic C viral hepatitis. As it comes to our knowledge, till now, there were published only 4 cases of generalized seizures that occurred during treatment with IFN alpha for chronic C viral hepatitis. The physiopathology of this complication is unknown. Generalized seizures can be reasonable due to IFN-alpha therapy, as long as the patient didn't have any seizure history, or other factors, which can develop seizures. Neurological examination, EEG and brain scan were normal. The recurrence of these seizures was absent stopping IFN-alpha therapy without any other seizure treatment. PMID- 12089979 TI - Abdominal pregnancy. Case report, review of the literature. AB - Abdominal pregnancy (AP) is a rare form of ectopic pregnancy (EP). It is a medical emergency because of high maternal and fetal morbidity and mortality. Early diagnosis is essential. Surgery is indicated as soon as the diagnosis is made. We present a case with AP in the third trimester with malformed fetus and left ovarian tumour. The fetus has been extracted at 33 weeks gestational age; the placenta and the ovarian tumour have been removed five weeks later. Pathology showed corpus luteum with hemorrhage, serous cystadenoma, follicular cyst. The fetus died, the woman successfully recovered. PMID- 12089980 TI - [Association between phakomatosis and neoplasia in children pathology. IV Pediatric Clinic experience]. AB - The risk to developing a neoplasm is increased when associated to a patient phakomatosis (Recklinghausen neurofibromatosis, Bourneville's tuberous sclerosis). We analysed 6 cases with phakomatosis and tumours, admitted in the Department of Oncopediatry, between 1993-1998; five of these children had neurofibromatosis and one Bourneville's disease. The associated tumours were hematologic malignancies (juvenile myeloid chronic leukemia) and solid tumors (rhabdomyosarcoma, hepatic carcinoma, CNS tumour, NHL optic glioma). The diagnosis was confirmed by microscopic examination of the bioptic material in all cases. Tumoral staging was performed by clinics, biology and imagistic investigations. All cases had extensive and aggressive tumours at the moment of diagnosis, We noticed a poor response and an early relapse after chemotherapy. A special follow-up and a different management has to be established for the patients with phakomatosis, in order to have a good oncological prophylaxis. PMID- 12089981 TI - [Implications of oxidative stress in senescence]. AB - After a short review of the literature concerning the paradox of oxygen and the participation of the oxidative stress created by the prevalence of its reactive radical species over the antioxidant enzymes, the main experimental arguments favoring the hypothesis of the implication of free radicals in senescence will be exposed. The amplification of degrading processes of lipid peroxidation and altering of the protein thiol groups in the presence of reactive oxygen species will be outlined. Both the aging of mitochondria due to the alteration of the electron transport chain and the oxidative attack on the mitochondrial DNA and the activation of the apoptosis as programmed cell death phenomenon in the presence of free radicals of oxygen, nitrogen and carbon will be given a particular interest. Finally, the implication of radical species in the production of some cardiovascular degenerative diseases of elderly patients will be discussed, together with the protective role of antioxidant compounds and diet, as factors that confine the progressive degradation of the senescent human body. PMID- 12089982 TI - Therapeutical aspects of the plasmocytoma localized at the level of head and neck. AB - The authors present a study on 7 cases of extramedullary plasmocytoma localized in E.N.T. area, hospitalized and solved in the E.N.T. Department of the the "Sf. Spiridon" Universitary Hospital, in the last 15 years, between 1983-1997. We insist on the last solved case, presenting a double isolated localization, a laryngeal and a rhinosinusal one. We consider that the most indicated treatment in the isolated extramedullary plasmocytoma, is the radiosurgical therapy with a good rate of survival. PMID- 12089983 TI - Barraquer-Simons syndrome. Report of a case and review of the literature. AB - Among lipodystrophies, the Barraquer-Simons syndrome or "parital lipodystrophy of the cephalothoracic type" is a rare condition of unknown origin characterized by a progressive atrophy of the subcutaneous fat of the upper part of the body. We report a 34-year-old woman with progressive loss of subcutaneous fat during 20 years, first affecting the face and spreading to the upper part of the body, equally presenting hypertrophy of the lower body fatty tissue. An overview of this rare syndrome is made. PMID- 12089984 TI - [Hysteroscopic endometrial ablation validation among conservative treatment methods of menorrhagia]. AB - Hysterectomy begins to fail as an absolute treatment for menorrhagia, as new medical and conservative treatment occurs. The author analyzes literature data of efficiency of hysteroscopic endometrial ablation compared with medical treatment, as well as among different hysteroscopic techniques. In conclusion, considering the expansion of hysteroscopic methods, gynaecologists should consider revising the treatment option in dysfunctional uterine bleeding, and offer to the patients the option of endometrial ablation. PMID- 12089985 TI - [CAD/CAM technology-elaboration of electronic model through optical impression in the fabrication of the dental prostheses by a computer assisted device]. AB - In modern CAD/CAM technology of dental prosthetics, the "optical impression" taken by special video cameras deploying 3D-recording of the oral situation (Cerec 1, Cerec 2) initiates the computer-generated production of a digital model. The computer receives and renders the digital data obtained through "optical impression" i.e. data acquisition not as a "negative" of the oral situation but as a "positive" copy of it, namely as a digital model in a virtual environment which can be displayed on the screen and be further used in the fabrication of dental prostheses by means of a computer-assisted device (CAM or CIM). PMID- 12089986 TI - [History of Romanian surgery]. PMID- 12089987 TI - [Genetic susceptibility to cancer]. AB - Hereditary predisposition is a common trait of many cancers. 15 to 20 percent of all cancers occur in individuals who have inherited a single gene alteration being members of families where multiple persons carry a high risk of developing cancer. Other than these so-called high penetrance genes which confer elevated risks of cancer development, there are many other genes that generate less dramatic but clinically important risks of cancer, often only if associated to specific exposures. PMID- 12089988 TI - [Cardiovascular risk factors in the elderly]. AB - The paper presents a review of recent findings about cardiovascular risk factors in elderly. It becomes important to know that the cardiovascular risk factors we look for in adulthood change in elderly patients. However, we consider that the cardiovascular risk factors in elderly remain hypertension, smoking status, hypercholesterolemia, diabetes mellitus and obesity. Recent studies proved that in elderly high levels of cholesterol are much less found than in adults as well as smoking status. Elderly has specific risk factors: high levels of iron and basic tachycardia. Other possible risk factors are: high levels of homocysteine, low plasmatic levels of HDL-cholesterol, high levels of lipoprotein-A and some coagulation factors. PMID- 12089989 TI - Neurofibromatosis. Nosological considerations. AB - The neurofibromatosis (NF) represent a set of conditions having different clinical manifestations, prognosis and inheritance. It has been presented--on clinical grounds--seven types of NF, but for only two of these National Institute of Health Consensus Development Conference (NIHCDC) advent a set of diagnostic criteria. The genes responsible for NF1 and NF2 were mapped to the long arm of chromosome 17 (17q11.2) and respectively 22 (22q11.2), and their protein product (neurofibromin and respectively merlin or schwanomin) was identified. Recent studies are proved that NF1 and NF2 genes act as a tumour suppressor gene. Up to now, only a limited number of mutations in these genes have been characterized but even in these cases the genotype/fenotype correlation has not provided enough information to allow speculation on the etiologic role NF1 or NF2 mutations might play in the variant forms of NF. Further studies are required to elucidate the genes functions and mutation spectrum. This should provide a framework for the molecular classification and diagnosis and the development of new therapy for NF. PMID- 12089990 TI - [Fluoroquinolones. Drug interactions]. AB - This review summarizes clinically relevant drug-drug interactions for fluoroquinolones: antiacids containing aluminum and magnesium salts, iron or zinc preparations, sucralfate, cimetidine, ranitidine, warfarina, cyclosporin, rifampin, oral contraceptive steroids, benzodiazepine, probenecid, beta-lactam antibiotics, nonsteroidal anti-inflammatory drugs, metronidazole, theophylline, caffeine. PMID- 12089991 TI - [Emergence of transmissible disorders, a continuous process--a new type of viral meningoencephalitis]. AB - In the interval 1994-1999, in Australia, Malaysia and Singapore, epizootic and epidemiological episodes of meningoencephalitis and severe acute respiratory syndromes were reported. Highly lethal in horses, swine and humans, the episodes were proved to be caused by the "new" viruses Hendra (HeV) and Nipah (NiV). At the same time three "new" viral agents have been isolated: Lyssavirus, Menanglevirus and Tupaia paramyxovirus. The intense contemporary circulation of people, animals and food products together with changes in human ecosystem favor new relations between humans and the "natural reservoirs" of biologic agents with a pathogenic potential for domestic and peridomestic animals and humans. PMID- 12089992 TI - [Childhood toxoplasmosis today]. AB - The comparative study of two groups of children suffering from acquired or congenital toxoplasmosis, being in hospital during two different decades (1979 1985--39 children and 1991-1997--117 children) proved: the fact that atypical or subclinical manifestations of the acquired toxoplasmosis (54%) draws our attention to the systematic research of the infection with Toxoplasma gondii, especially at the groups with increased risk now that the frequency of the diseases generating immunodeficiency; the increased frequency of congenital toxoplasmosis (5% in the first group, 8% in the second group) and especially of the invalidating of screening tests to the pregnant women; the therapeutic arsenal limited by the rate price/toxicity/medical resistance, must be individualized for each case, taking into account the increased number of other associated diseases (chronic hepatitis B 7%, TB 17%, HIV infection 5%); Rovamicine treatment at the second group had similar results to those obtained by administrating Pirimetamine--Trimetoprim to the children from the first group. PMID- 12089994 TI - [Gastroesophageal reflux disease and Helicobacter pylori]. PMID- 12089993 TI - [Prognosis and treatment of membranous glomerulonephritis-a 5-year prospective study]. AB - The aims of the study were to describe the clinical, pathological and biological features of membranous GN and to prospectively evaluate the relationships between individual negative prognostic factors--type of therapy and outcome. Between 1993 1998, 13/150 (8.7%) consecutive patients with renal biopsy had membranous GN (M = 62%, age = 42.5 +/- 14.5 years). Main (major) findings in these patients were: asymptomatic proteinuria--23.1%, heavy proteinuria (> 10 g/day)--33.3%, microscopic hematuria--53.8%, increased plasma creatinine levels--33.3%, hypertension--23.1% cases. 60% of the patients with nephrotic proteinuria had an underlying cause (infection, malignancy, immune-mediated systemic diseases). 40% of the patients with nephrotic proteinuria had 0 or less than 2 negative prognostic factors (without any of the recognized severe morphological changes). The following differentiated treatment protocols were applied: no treatment for asymptomatic proteinuria (group A), i.v. methyl-prednisolone boluses + prednisone 1 mg/kgc/day 3 months for those patients with few negative prognostic factors (group B), and steroids (as above) + cyclophosphamide (2 mg/kgc/day 3 months) or the Ponticelli regime in patients with important risk factors (group C). Outcome after a median follow-up period of 24 months was: complete remission in all cases from groups A + B (with only one exception were the underlying cause was breast malignancy); in group C in 75% of the subjects a complete or partial remission (proteinuria < 1 g/day) was obtained. Only one case progressed to chronic renal failure. There were no secondary effects from corticoids or immunosuppressive therapy. CONCLUSIONS: In membranous GN treatment should be tailored to the presence and type of negative prognostic factors. Even in high-risk patients combined steroids and immunosuppressive therapy determines a favorable outcome in 75% of the cases, without severe adverse effects. PMID- 12089996 TI - [Hanover intensive score. Study on 218 cases ]. AB - We have performed a prospective study (1990-1996) in our intensive care unit in Hospital "C.I. Parhon" Iassy. Our study has 218 critical patients with ARDS and MSOD. For all of them we have used a new scoring system HIS (Hanover Intensive Score) for: the evaluation of patients with ARDS that have the risk to develop MSOD; determine the prognosis in different moments of evolution. In our opinion this score has the following advantages: it is easy to apply; it can determine the prognosis and the probable status of the patient at the moment of outcome; it can be used in our country with the technical that we have and we propose this score for his performances. PMID- 12089995 TI - [Body composition in amenorrheic and eumenorrheic underweight patients]. AB - Relationship between weight and hypothalamic-pituitary-ovarian axis has been long time studied. Underweight female patients can have or not disturbances of gonadal system and the importance of the weight or body composition in the mechanisms of these abnormalities is not clear. In our study, we have investigated body composition in 15 underweight female patients. Five patients have had anorexia nervosa, with amenorrhea (group AN), and other 10 patients were eumenorrheic, but with low body weight (hypoanabolic syndrome--group HA). Bioelectrical impedance method was used to investigate body composition. Total body water (TBW), lean mass (LM), and fat mass (FM) were determined. Nutritional status was also assessed using body mass index (BMI). The mean body weight was significantly decreased (p = 0.05, 36.6 +/- 4.68 Kg versus 44.1 +/- 4.67 Kg) in group AN as compared to group HA. Fat mass was very low in group AN (9.45% of body weight versus 11.3% of body weight in group HA), but there was not significant difference. Lean mass was significantly decreased in group AN (33.24 +/- 2.37 Kg) in comparison with group HA (39.3 +/- 4.06 Kg). Correlations between anthropometric indices were studied. Positive significant correlations were obtained between body weight and lean mass, total body water and basal metabolism in both groups. In conclusion, our study reveals that body weight has a relative importance in ovarian axis dysfunction, and the most important of body compartments seems to be lean mass. PMID- 12089998 TI - [Achalasia in children: Diagnostic opportunities and therapeutical options]. AB - Achalasia is a functional disorder of the distal esophagus, which fails to relax during swallowing. Although being rare, this disorder is the source of a significant morbidity, including developmental disorders in children. Several therapeutical options are available: myorelaxants administration, esophageal dilations and surgery--distal esophageal myomectomy followed by an antireflux procedure. The paper introduces our experience in treating a number of 5 children between 1991 and 1998. PMID- 12089997 TI - [Cephalic duodenopancreatectomy with pyloric preservation in the treatment of pancreatic cancer]. AB - Is the application of DPCPP in the treatment of pancreatic neoplasia a good reason? We have analysed 30 patients with cephalic duodenopancreatectomy (DPC) for biliopancreatic neoplasia between 1995-1999 in Ist Surgical Clinic of Iassy (13 with pyloric preservation). The indications were:--cephalic pancreatic neoplasia (adenocarcinoma--4 cases (one with cephalic chronic pancreatitis on the intraoperative microscopical examination);--Vater ampulloma (7 cases);--inferior common biliary duct (CBD 1 case);--duodenal adenocarcinoma (1 case). In the same time was operated 265 biliopancreatic diseases (203 mechanical jaundice with 132 neoplastic jaundice). RESULTS: --Better early postoperatively status of the patients--DPCPP does not give better prognosis;--there are necessary some technical skills to depase the important phases of DPCPP. PMID- 12089999 TI - [Evaluation of sepsis prognosis using Saps II]. AB - The simplified acute physiological score (SAPS II), the only valid score in sepsis according Pilly (1997) includes 17 variables: 12 physiological variables, age, type of admission and 3 variables that reffer to the background diseases: AIDS, neoplasm and haematologic malignant diseases. SAPS II was used in 30 patients with sepsis. The evaluation for each variable were between 0-26 points. Our data suggest that values over 0.552 coincided with death in 12 patients and values below 0.552 coincided with a favourable course in 18 patients. PMID- 12090000 TI - Apoptotic cell death: from worms to wombats ... but what about the weeds? PMID- 12090001 TI - Organelle-specific death pathways. PMID- 12090002 TI - Caspases--at the cutting edge of cell death. PMID- 12090003 TI - Telomeres, telomerase and cellular immortalization. PMID- 12090004 TI - Back from the brink: plant senescence and its reversibility. PMID- 12090005 TI - Senescence and cell death in Brassica napus and Arabidopsis. PMID- 12090006 TI - Cytokinin and its receptors. PMID- 12090007 TI - Ethylene-triggered cell death during aerenchyma formation in roots. PMID- 12090008 TI - Targeted cell death in xylogenesis. PMID- 12090009 TI - Abscission and dehiscence. PMID- 12090011 TI - Tissue transglutaminase in cell death. PMID- 12090010 TI - Regulation of cell cycle arrest and cell death--alternative responses to DNA damage. PMID- 12090013 TI - Tumour cell death. PMID- 12090012 TI - Pathways used by adenovirus E1B 19K to inhibit apoptosis. PMID- 12090014 TI - Controlling apoptosis: implications for carcinogenesis? PMID- 12090015 TI - Type I interferons inhibit the resolution of chronic inflammation. PMID- 12090016 TI - Death of plant cells and their contribution to rumen function. PMID- 12090017 TI - Postscript: some concluding observations and speculations. PMID- 12090018 TI - Modulation of death receptor signalling. PMID- 12090019 TI - The role of the Bcl-2 family in the modulation of apoptosis. PMID- 12090020 TI - The role of granzymes and serpins in regulating cell growth and death. PMID- 12090022 TI - Regulation of apoptosis by cell metabolism, cytochrome c and the cytoskeleton. PMID- 12090021 TI - Mitochondria and cell death: a pore way to die? PMID- 12090023 TI - Feeding of red palm oil-supplemented diets to rats may impact positively on vitamin A status. AB - The impact of feeding of dietary palm oil supplements on plasma vitamin A profile was investigated in animals. Four-week-old Wistar albino rats (n = 8 per group) were maintained for 28 days on standard rat food (4.7% fat by weight) supplemented (10%, 20% and 30% by weight) with red palm oil (RPO) and refined palm olein (REFPO). Plasma beta-carotene and vitamin A concentrations of rats fed RPO-based diets were higher than in rats fed REFPO and control diets. Animals fed 30% RPO-containing diets had lower plasma beta-carotene concentrations than those fed 20% RPO-containing diets. The results suggest that consumption of palm oil in moderate amounts enhances growth of tissues and bioavailability of beta-carotene, which may combat vitamin A deficiency in developing countries, in view of the fact that performed vitamin A in animal products (namely meat, liver, eggs and fatty fish) is out of the reach of economically deprived people. PMID- 12090024 TI - Soluble and insoluble oxalate content of mushrooms. AB - Six different cultivars of edible fungi were bought from a local supermarket in Uppsala, Sweden and three different cultivars of wild growing fungi were harvested from the forest adjoining the city. The edible portions of the raw fungi were then analysed for total and soluble oxalate content. A portion of each fungi was then cooked by first boiling in a small amount of water followed by frying in a small amount of butter. The total oxalate content of the raw, commercially grown mushrooms ranged from 58.9 to 104.1 mg/100 g DM while the values for the total oxalate content of the cooked mushrooms ranged from 48.9 to 124.6 mg/100 g DM. Apart from cultivars ostronskivling and piopino the cultivated mushrooms contained mostly insoluble oxalates. Mushrooms harvested from the forest contained only soluble oxalate with the levels ranging from 29.3 to 40.2 mg/100 g DM in the raw tissue. Cooking marginally lowered the soluble oxalate content of these mushrooms. The levels of soluble and insoluble oxalates of all the mushrooms analysed were low compared to other common oxalate-containing vegetables. PMID- 12090025 TI - Lower diet glycaemic index in African than South Asian men in Trinidad and Tobago. AB - In Trinidad and Tobago (T&T), diabetes is over two times more common in South Asian (SA) than African (AF) men, with a smaller difference in women. In the USA, high diet glycaemic index (GI) was associated with increased risk of developing diabetes. In T&T, dietary intake data are unavailable. The objective of this study was to conduct a pilot survey of the dietary habits of 50 AF and 50 SA subjects in order to develop a valid nutrition survey instrument for use in T&T. We hypothesized that diet GI would be lower in AF than SA subjects. We collected 24 h dietary recalls from a sample of adults from T&T (26 AF and 33 SA women, and 17 AF and 25 SA men), excluding subjects of mixed or other ethnic backgrounds and those with diabetes or other chronic disorders. AF women were of similar age, but taller and heavier than SA women. There was no significant difference in diet between AF and SA women. AF men were taller than SA men, but of similar age (34 +/- 2 years versus 33 +/- 2 years) and body mass index (24.1 +/- 0.8 kg/m2 versus 23.3 +/- 0.9 kg/m2, respectively). Intakes of energy, fat, protein, carbohydrate and fibre, and diet glycaemic load did not differ significantly, but AF men had a lower diet GI than SA men (56 +/- 1 versus 59 +/- 1, P < 0.05). Although not conclusive, these results are consistent with previous data suggesting that diet GI may contribute to the development of type 2 diabetes. The data collected during this survey will be useful in developing a food frequency questionnaire for use in T&T to allow this hypothesis to be tested more rigorously in this population. PMID- 12090026 TI - Influence of dietary carbohydrates and glycaemic response on subjective appetite and food intake in healthy elderly persons. AB - Increased satiety and decreased food intake are reported following the consumption of low glycaemic index (GI) foods, which gradually increase blood glucose. This observation, however, is not uniformly supported and few studies have examined the impact of different GI foods on satiety and intake in the elderly. After an overnight fast, 10 men and 10 women (aged 60-82 years) consumed similar amounts of available carbohydrate as high (glucose drink or potatoes) or low (barley) GI foods or a non-energy placebo drink on four mornings. Blood glucose and subjective appetite were measured throughout a 120 min post-ingestion period, followed by consumption of an ad libitum lunch. Differences in plasma glucose after test food ingestion (glucose > potatoes > barley > placebo; P < 0.03) did not predict subjective appetite or lunch intake. Potatoes increased subjective satiety the most, followed by barley, then glucose, which trended towards greater satiety than placebo. Potatoes led to less hunger than placebo (P = 0.0023) and less prospective consumption than the other three foods (P < 0.0083), and potatoes and barley led to greater fullness than glucose and placebo (P < 0.0001). Lunch intake was decreased, compared with placebo (502 +/- 47 kcal, P < 0.031), by potatoes (405 +/- 40 kcal) and barley (441 +/- 41 kcal); however, only potatoes (41.9 +/- 12.3%) led to greater compensation at lunch for test food ingestion compared with glucose (11.9 +/- 9.5%, P = 0.016). These results suggest that elderly subjects are sensitive to the effects of different foods on subjective appetite and food intake, and that the GI of the foods tested did not predict their effects on satiety and food intake. PMID- 12090027 TI - Influences of dietary protein levels and phytate contents on zinc requirement in rats. AB - This study investigates whether the effects of increased dietary levels of soy protein on zinc-deficient rats are due to the dietary protein content. Rats were fed two levels of demineralized soy protein (DP) diets and two levels of egg albumin (EA) diets including 7 ppm zinc (Experiment 1). Growth was depressed in rats fed a 20% DP diet (0.43% phytate) but not in those fed a 10% EA diet containing 7 ppm zinc. Zinc concentrations in the serum, femur and kidney were lower in rats fed the 20% DP diet than those fed a 10% DP (0.20% phytate) diet, and they were also lower in rats fed the 10% EA diet than those fed a 5% EA diet. Zinc concentrations in the serum and femur of rats fed the 10% DP diet or the 20% DP diet were decreased compared with those fed the 5% EA diet or the 10% EA diet, respectively. Under zinc-deficient conditions (< 0.4 ppm Zn), the survival time shortened (Experiment 2) and the zinc concentration in the serum and femur decreased faster (Experiment 3) in rats fed the 10% EA diet compared with those fed the 5% EA diet. The survival times and time courses of these parameters show that the zinc requirements of rats increased with the dietary protein level. The increased zinc requirement of rats that accompanied increasing dietary soy protein was due to the dietary protein content as well as the dietary phytate content. PMID- 12090028 TI - Prevalence of risk factors to coronary heart disease in an Arab-American population in southeast Michigan. AB - Although significant advances have been made in the area of cardiovascular disease, few studies have targeted ethnic groups. There is a large and growing Arab-American (AA) population living in Southeast Michigan, whose risk of cardiovascular disease may be on the increase. The objective of this study was to evaluate the prevalence of cardiovascular disease risk factors and associated behavioral factors in an AA community with a large population of emigrants, subjected to significant lifestyle changes. Three hundred and fifty-two AA living in Southeast Michigan, mostly from the Middle East, were screened to determine their eating and smoking habits, body mass index (BMI) body fat analysis, blood pressure, and complete lipid profiling. Overweight was defined as a BMI greater than or equal to the 85th percentile value for age- and sex-specific reference data from the third National Health and Nutrition Examination Survey (NHANES III). Correlation analysis was used to examine factors associated with being overweight, with adjustment for age and sex. Blood cholesterol concentrations were compared with published data for Arabs from the Middle Eastern countries. The overall prevalence of being overweight in subjects aged 35 and older was significantly higher than NHANES III reference data (Men, 27.7% (95% confidence interval, 21.8-34.5); women, 33.7% (95% confidence interval, 27.9-40.1)). A mean cholesterol concentration of 210 +/- 4 mg/dl was observed in those over the age of 40. The mean high-density lipoprotein (HDL)-cholesterol levels for men and women were 38 and 48 mg/dl, respectively. Greater than 54.6% of all subjects had a total cholesterol:HDL ratio > 4.5. Although being overweight and obesity were prevalent in this population, the mean BMI for men was 25.7 +/- 0.34, compared with 27 +/- 0.58 for women. Increased BMI was significantly correlated (P < 0.01) with increased blood pressure, increased glucose levels, increased total cholesterol and decreased HDL-cholesterol levels (P < 0.01). Elevation in risk factors to cardiovascular disease is prevalent in this population and indicates a need for programs targeting primary prevention of obesity in men and women. These results, which could be attributed in part to lifestyle changes typical of most emigrant populations, suggest an increase in the risk for developing cardiovascular disease. In addition, this study provides a basis for future intervention to improve the health of this population. PMID- 12090029 TI - Cleaning frequency and the microbial load in ice-cream. AB - This study investigates the efficacy of a 62 h cleaning frequency in the manufacturing of ice-cream. Various product and product contact surfaces were sampled progressively throughout the time period between cleaning cycles, and analyzed for microbial growth. The coliform and standard plate counts (SPC) of these samples did not vary significantly over time after 0, 24, 48, or 62 h from Cleaning in Place (CiP). Data for product contact surfaces were significant for the SPC representing sample locations. Some of the variables in cleaning practices had significant influence on microbial loads. An increase in the number of flavors manufactured caused a decrease in SPC within the 24 h interval, but by the 48 h interval the SPC increased. More washouts within the first 24 h interval were favorable, as indicated by decreased SPC. The more frequently the liquefier was sanitized within the 62 h interval, the lower the SPC. This study indicates that food safety was not compromised and safety practices were effectively implemented throughout the process. PMID- 12090030 TI - Loss of residual cyanogens in a cassava food during short-term storage. AB - Residual cyanogens in gari, the most popular cassava food in West Africa, is implicated in the causation of tropical ataxic neuropathy. Gari is eaten by soaking its granules in cold water or by adding boiling water to make a food called eba. This study was conducted to determine whether loss of the residual cyanogens in gari during short-term storage and when gari is made into eba will reduce dietary cyanide load in consumers. Fifteen samples of gari, nine roasted from cassava mash fermented for at least 4 days (type A) and six roasted from cassava mash fermented for only 1 day (type B), were stored for 4 weeks. Free cyanide, linamarin, and cyanohydrin in gari and in eba made from the gari were determined at weekly intervals for 4 weeks. Free cyanide was absent in all samples of gari. Mean cyanohydrin dropped from 8.4 mg HCN Eq/kg dry weight to 4.6 mg HCN Eq/kg dry weight in type A gari, while it dropped from 3.0 mg HCN Eq/kg dry weight to 1.3 mg HCN Eq/kg dry weight in type B gari. Mean linamarin dropped from 6.6 mg HCN Eq/kg dry weight to 2.8 mg HCN Eq/kg dry weight in type A gari, while it dropped from 1.7 mg HCN Eq/kg dry weight to 0.4 mg HCN Eq/kg dry weight in type B gari. Loss of linamarin and cyanohydrin was significant at P < 0.001 for type A gari and at P < 0.002 for type B gari when the weekly levels were compared with initial values. When gari was made into eba, 36% of cyanohydrin and 47% of linamarin were lost from type A gari, while 38% of cyanohydrin and 5% of linamarin were lost from type B gari. Loss of linamarin and cyanohydrin when gari was made into eba was significant for both types of gari at P < 0.001. This study shows that the loss of cyanohydrin and linamarin in gari during short-term storage and when gari is made to eba will reduce dietary cyanide load in consumers. PMID- 12090031 TI - Deterioration of natural antioxidant species of vegetable edible oils during the domestic deep-frying and pan-frying of potatoes. AB - In the present work, virgin olive oil, sunflower oil and a vegetable shortening were used as cooking oils for the deep-frying and pan-frying of potatoes, for eight successive sessions, under the usual domestic practice. Several chemical and physicochemical parameters (acidic value, peroxide value, total polar artefacts, total phenol content and triglyceride fatty acyl moiety composition) were assayed during frying operations in order to evaluate the status of the frying oils, which were found within expected ranges similar to those previously reported. The oil fatty acids were effectively protected from oxidation by the natural antioxidants. The frying oil absorption by the potatoes was quantitated within 6.1-12.8%, depending on the oil type and the frying process. The retention of alpha- and (beta + gamma)-tocopherols during the eight fryings ranged from 85 90% (first frying) to 15-40% (eighth frying), except for the (beta + gamma) tocopherols of sunflower oil, which almost disappeared after the sixth frying. The deterioration during the successive frying of several phenolic species present in virgin olive oil is reported for the first time. The retention of total phenolics ranged from 70-80% (first frying) to 20-30% (eighth frying). Tannic acid, oleuropein and hydroxytyrosol-elenolic acid dialdeydic form showed remarkable resistance in all frying sessions in both frying methods, while hydroxytyrosol and hydroxytyrosol-elenolic acid were the faster eliminated. The deterioration of the other phenolic species account for 40-50% and 20-30% for deep-frying and pan-frying, respectively, after three to four frying sessions, which are the most usual in the household kitchen. Deep-frying resulted in better recoveries of all the parameters examined. The correlation of the deterioration rate of the phenolic compounds and tocopherols during frying is discussed and the nutritional aspects of the natural antioxidant intake, through the oil absorbed by the potatoes, are evaluated. PMID- 12090032 TI - [Reduced-intensity allogeneic hematopoietic stem cell transplantation (mini transplantation) for solid tumors]. AB - There are accumulating evidence of immunological therapeutic effect induced by reduced-intensity allogeneic hematopoietic stem cell transplantation (mini transplantation) for solid malignancies. The reduced toxicity of a mild preconditioning regimen in mini-transplantation has facilitated its application to patients with various solid tumors. The so-called graft-versus-tumor (GVT) effect usually appears when graft-versus-host disease develops. Therefore, it is suggested that the fundamental mechanism of the GVT effect is a reaction by T lymphocytes against various tumor-associated alloantigens of the tumor cells. Although mini-transplantation appears effective in some patients with renal cell carcinoma or breast cancer, it is still unclear whether the therapeutic mechanism could work on other solid tumors. Well-designed prospective clinical trials are warranted in order to determine the role of mini-transplant in the therapeutic strategy for solid tumors. PMID- 12090033 TI - [The diagnostic imagings of liver metastases]. AB - The liver is an organ susceptible metastases. Malignant neoplasms of many organs frequently metastasize to the liver, particularly colon malignancies. Most metastases enter the liver via the blood circulation, but metastases through lymphatic vessels and the peritoneum are also common. The morphologies of liver metastases and findings on diagnostic images vary considerably. Numerous modalities are available for diagnostic imaging of liver metastases: ultrasonography, computed tomography (CT), magnetic resonance imaging (MRI), angiography, nuclear medicine, and others. Ultrasonography is the simplest, most non-invasive, and cheapest, but relies on the skill of the operator. CT and MRI are also non-invasive, and the sensitivities of these techniques have recently been improved with the development of multidetector CT, contrast agents for MRI that specifically accumulate in the liver, and other advances. Moreover, positron emission CT using fluorine-18-deoxyglucose (FDG-PET) displays a high sensitivity. Angiography is an invasive modality, but is adopted with CT during arterial portography (CTAP) and CT during hepatic arteriography (CTHA), or for the purpose of interventional radiology. Rational selection of appropriate modalities for a given purpose requires familiarity with the characteristics of each modality. In the present paper, we describe the morphologic characters of liver metastases and investigate the associated characteristics and usefulness of each modality. PMID- 12090034 TI - [The impact of hepatic resection on metastatic colorectal cancer]. AB - Although liver resection is the accepted treatment for patients with metastatic colorectal cancer to the liver, there remains some controversy as to the criteria for patient selection and its impact on the recurrent disease following the operation. One hundred and sixteen patients underwent liver resection for metastaic colorectal carcinoma over the last 23 years at our institution. The actual survival was 50.5% at 5 years, and 38.6% at 10 years, excluding the 30-day operative mortality rate of 1.7%. Positive hepatic lymph nodes, extrahepatic disease, number of tumors (4 or more), and bilobar distribution of the diseases were strongly associated with poor outcome. These prognostic factors were also documented as the major prognostic determinants by 21 consecutive articles we reviewed. Among 52 (45.6%) patients with hepatic recurrence, patients who were acceptable surgical risks and had no extrahepatic diseases or a limited number (one or two) of lung metastasis were candidates for the repeat resection. Consequently, 31 patients underwent repeated hepatic resection. Their survival rate was 53.9% at 3 years and 32.3% at 5 years, with no mortality. This outcome is markedly better than that of untreated patients with an estimated survival of 3 to 24 months. Thus, an aggressive approach to surgical treatment is feasible with low risk and the potentially curative strategy even for recurrent hepatic metastases, providing the opportunity to achieve 5 year survival. PMID- 12090035 TI - [Microwave coagulation therapy for unresectable colorectal metastatic liver tumor]. AB - INTRODUCTION: We evaluate the utility of microwave coagulation therapy (MCT) for the patients with unresectable liver metastasis from colorectal cancers. METHODS: Forty-four patients unresectable synchronous colorectal hepatic metastasis between January 1, 1989 and December 31, 2000, were enrolled in the present study. The patients courses were followed until March, 2002. To evaluate the efficacy of MCT for unresectable liver metastasis from colorectal cancers, a retrospective comparative study was done between a MCT with CT (hepatic arterial trans-infusion chemotherapy and/or systemic chemotherapy) group and a CT group. The background factors were examined included the maximum tumor diameter, the number of liver metastasis and disease of other metastatic sites, which influenced the outcome. RESULTS AND CONCLUSION: The survival curves of the patients with unresectable synchronous colorectal hepatic metastasis undergoing MCT with CT and CT alone were significantly different (p = 0.03). No significant differences were found in the background factors that had a prognostic influence. It is confirmed that using MCT can prolong the survival rate for the patients with unresectable synchronous colorectal hepatic metastasis. PMID- 12090036 TI - [Treatment for patients with liver metastasis from colorectal cancer--expansion of hepatic arterial infusion therapy]. AB - The treatment of patients with liver metastasis from colorectal cancer was evaluated. Although surgical resection was the most effective therapy and 5-year survival was 30%, more than two-thirds of the patients were judged unresectable at the beginning of the treatment. Those whose metastasis was judged to be unresectable received hepatic arterial infusion (HAI) of 5-FU therapy. In a comparison of the survival period, patients received more than 15 g of 5-FU were found to have a statistically significant longer survival than those who received less than 15 g. Even though the patients who revealed a PR or CR response with HAI did not survive more than 4 years, we began surgical resection in these patients after HAI therapy. As a result, the 5-year survival rate of patients whose metastasis was unresectable and who underwent surgical resection after successful HAI was 30% which was equal to the result of the patients judged resectable at the beginning of the treatment. Such treatment may become the fundamental treatment for unresectable liver metastasis in the near future. PMID- 12090037 TI - [Systemic chemotherapy for advanced colorectal cancer with liver metastasis]. AB - We report the progress of systemic chemotherapy for advanced colorectal cancer with liver metastasis. It must be noted that the purpose of this treatment is to prolong the symptom-free period. Review of hepatic arterial infusion (HAI) compared with systemic chemotherapy for the treatment of unresectable liver metastases from colorectal cancer showed that was attained with HAI a much higher response rate and survival benefit than systemic chemotherapy. However, systemic chemotherapy has shown progress since that time. Regarding administration methods, continuous injection is better than bolus injection for 5-FU. New modulators of 5-FU have also became available, such as leucovorin, CPT-11, and I OHP. Futhermore, many studies of 5-FU-based combination therapy have shown that the mean survival time (MST) and response rate (RR) are now close to those of HAI. Finally, the combination with HAI with systemic chemotherapy using CPT-11 resulted in the highest RR of 74%. Further trials of such combination therapy will be performed in the future. PMID- 12090038 TI - [Evaluation of preoperative simultaneous radiochemotherapy combined with carboplatin and tegafur.uracil]. AB - We performed preoperative radiochemotherapy including local irradiation with total dose of 40 Gy along with concomitant combination chemotherapy with carboplatin and tegafur.uracil for 11 head and neck squamous cell carcinomas and evaluated the clinical and histopathological effects as well as the side effects. Seventy-three percent of patients accomplished significant clinical effects equal to or more than PR and significant histopathological effects in surgically resected tissues. Side effects were seen in only one patient with severe hypoleukocytemia, no patients with renal dysfunction, and 34% of patients with severe oral mucositis. These results suggest that our radiochemotherapy may be useful for preoperative treatment for patients with locally advanced head and neck squamous cell carcinomas. PMID- 12090039 TI - [Assessment of post-administration body distribution of toremifene and tamoxifen, and their administration regimens]. AB - Toremifene is an anti-estrogenic drug like tamoxifen. We assessed the body distributions after administration of toremifene and tamoxifen in order to evaluate their treatment regimens by measuring the concentrations in tissues. It is known that, after toremifene (TOR) or tamoxifen (TAM) is consecutively administered to breast cancer patients, TOR or TAM and their main active N desmethyl-metabolites (TOR-1 or TAM-1) are detected in sera, tumor tissues, and lymph nodes. Accordingly, after we administered toremifene or tamoxifen to primary breast cancer patients previous to surgery, we measured the concentrations of TOR, TOR-1, TAM, TAM-1 in sera, tumor tissues, and lymph nodes. We found that the concentrations of TOR and TOR-1 in sera, tumors, and lymph nodes reached a peak about 2 weeks after administration of toremifene 40 mg. Likewise, the concentrations of TAM and TAM-1 in sera, tumors, and lymph nodes reached a peak about 2 weeks after administration of tamoxifen, although the peak levels were lower than those of TOR or TOR-1. The concentrations of TAM-1 in lymph nodes were significantly and positively correlated to the duration of administration of TAM, and it was predicted that the concentration of TAM-1 in lymph nodes would reach a steady state at more than 4 weeks after administration of tamoxifen. The concentrations of TOR and TOR-1 were higher in tumors and lymph nodes than in sera. Furthermore, the concentrations of TOR and TOR-1 were significantly higher than those of TAM and TAM-1 in sera and tumors, respectively. Moreover, the concentration in tissue increased in a dose-dependent manner with administration of toremifene 120 mg. There were no significant differences between breast cancers positive and negative for estrogen receptors, with regard to the concentrations of TOR and TOR-1 in either sera, tumors, or lymph nodes. In conclusion, it would be expected that treatment with toremifene might be more effective for breast cancer than that with tamoxifen. PMID- 12090040 TI - [Dihydropyrimidine dehydrogenase and thymidylate synthase in colon carcinoma]. AB - Dihydropyrimidine dehydrogenase (DPD) and thymidylate synthase (TS) in cancer are considered to play key roles in the sensitivity to 5-FU-based chemotherapy. We measured DPD activities and TS content in cancer and normal mucosa obtained from 59 patients with colorectal carcinomas and examined their relationship and clinicipathological relevance. DPD activity was significantly decreased in cancer compared to that in adjacent mucosa, irrespective of the clinicopathological parameters (p < 0.0001), while TS content was significantly increased in cancer (p < 0.0001). DPD activity in cancer correlated with that of the normal mucosa (r = 0.57, p < 0.001), but no correlation was found between TS content of normal mucosa and cancer. The DPD activity difference between cancer and normal mucosa was significantly correlated with difference in TS content (r = 0.70, p < 0.0001). These results suggest that the reduction of DPD activity is associated with the elevation of TS content, which may be connected with the development of 5-FU resistance and the effectiveness of biochemical modulation through the stabilization of TS inhibition with folic acid in colon carcinoma. PMID- 12090041 TI - [Questionnaire survey of systemic chemotherapy in colorectal cancer]. AB - The results of a questionnaire survey conducted by the Colorectal Surgical Club were analyzed to elucidate how clinical trials performed as well as points of concern in the treatment of colorectal cancer by colorectal specialists in Japan. Of the specialists, 92.2% responded that systemic chemotherapy was necessary for curA (Dukes C), in almost all cases administered postoperatively either orally or intravenously, and 96.6% said systemic chemotherapy was necessary for unresectable colorectal cancer, in almost all cases administered intravenously in combinations of anticancer drugs. The things considered most important for systemic chemotherapy were survival rate, effective rate, and safety, while the most for which most care was taken were patient consultations, periodical blood examination, and observance of drug application and dosage. PMID- 12090042 TI - [Chemotherapy by superselective intraarterial infusion of nedaplatin combined with radiotherapy for oral cancer]. AB - Nedaplatin (CDGP), which is a CDDP derivative, has been reported to be an effective anticancer agent for head and neck cancer. This study was performed to assess the feasibility of chemotherapy by superselective intraarterial infusion of nedaplatin (CDGP) in patients with oral cancers. Ten patients were treated with chemotherapy by superselective intraarterial infusion of CDGP combined with radiotherapy. The complete and partial response rates were 7/10 (70%) and 3/10 (30%), respectively. Nine patients showed grade 1-2 hematological toxicity including leukocytopenia and anemia. Thrombocytopenia of grade 4 was seen in only one patient. However, all the patients were free from renal dysfunction. From these results, it is suggested that this combination therapy might be quite effective and safe. Further study will be needed to determine its efficacy against oral cancer. PMID- 12090043 TI - [Non-randomized clinical study comparing chemotherapy plus radiotherapy with radiotherapy alone in neoadjuvant therapy for oral cancer]. AB - Neoadjuvant therapy plays an important role for organ preservation and survival rate in the treatment of oral cancer. We clinically compared the effect of neoadjuvant radiotherapy and chemoradiotherapy in patients with oral cancer. We retrospectively examined 47 patients diagnosed with oral squamous cell carcinoma who underwent neoadjuvant therapy followed by curative surgery in the oral and maxillofacial surgery department of Ehime University Hospital. We divided them into two groups: radiotherapy alone (24 cases) and chemoradiotherapy (23 cases). The patients in the radiotherapy group underwent irradiation of 32.6 +/- 5.0 Gy (mean +/- SD). The patients in the chemoradiotherapy group received a low-dose fraction of cisplatin (8 mg/mm2/day, 5 days a week; total dose: 139.4 +/- 67.1 mg) and 5-fluorouracil (300 mg/mm2/day, 5 days a week; total dose: 5,900 +/- 1,839.8 mg) combined with simultaneous irradiation of 31.0 +/- 3.2 Gy. None of the 24 patients had a complete response to radiotherapy alone and 12 (50%) had a partial response. Six (26%) of the 23 patients had a complete response to chemoradiotherapy and 12 (52%) had a partial response. The primary control rate (82.6%) to chemoradiotherapy was higher than that (67.5%) to radiotherapy alone although no significant difference was found. The 5-year survival rate was 64.3% in the radiotherapy group and 62.8% in the chemoradiotherapy group. The findings of the present study suggest that while the combination of radiation and cisplatin/5-fluorouracil in neoadjuvant therapy for oral cancer may not bring a significant benefit to improve survival rate, the primary local control rate is improved in comparison with radiotherapy alone. PMID- 12090044 TI - [A case of liver metastasis of breast cancer successfully treated with paclitaxel infusion into the hepatic artery: an attempt of once weekly regimen]. AB - On October 21, 1995, a 45-year-old woman underwent mastectomy (Kodama's procedure), preserving the thoracic muscles, based on the diagnosis of right breast cancer (T2aN1bM0). After surgery, she received 3 cycles of adjuvant chemotherapy (CAF) using anthracycline and other drugs. Thereafter she was orally treated with UFT for 2 years. She developed tumor recurrence in her liver 3 years after surgery. At that time, she was initially scheduled to undergo partial hepatectomy (including the tumor-affected area), but it was later judged that radical surgery was impossible due to the presence of multiple lesions in both lobes of the liver. A reservoir was therefore installed in the hepatic artery. Ten days after reservoir installation, intra-arterial infusion of paclitaxel (80 mg/m2), dissolved in 100 ml of physiological saline, was performed for one hour, using the installed reservoir. This treatment was administered once weekly for 3 consecutive weeks, followed by one week of rest. After 3 cycles of this regimen, the liver metastases had disappeared completely. At present, 2 years after the beginning of this therapy, the patient remains in a state of CR (complete response). Grade 2 alopecia and grade 1 peripheral neuropathy were the only adverse reactions observed. PMID- 12090045 TI - [A case of large-cell lung cancer successfully treated with docetaxel in combination with carboplatin and radiotherapy]. AB - A 64-year-old male was referred to our hospital in September, 2000 for further examination of an abnormal chest shadow discovered in a regular health check-up. Chest X-P and CT revealed a large tumor in the left upper lobe, in association with hilar lymphadenopathy and costal invasion. Serum CEA was increased, and lung biopsy revealed a large-cell carcinoma. We performed chemoradiotherapy before surgery. Radiation (total 48 Gy) and 3 courses of chemotherapy with docetaxel (60 mg/m2) in combination with carboplatin (AUC = 6,600 mg) resulted in a remarkable reduction in the size of the mass, to less than 50%, and normalized serum CEA. Left upper lobectomy, lymphadenectomy and costectomy were performed. However, no tumor cells were detectable in the ablative specimen pathologically. These findings suggest the efficacy of chemoradiotherapy including docetaxel with carboplatin in patients with large-cell lung cancer. PMID- 12090046 TI - [A case report of advanced gastric cancer responding to TS-1, a novel oral fluorouracil derivative]. AB - TS-1 is a new, oral anticancer agent composed of two modulators, gimeracil (CDHP) and oteracil potassium (Oxo) are mixed with tegafur in a ratio of 1:0.4:1. We report one case of advanced gastric cancer with lung and lymph node metastases that completely responded to TS-1. A 71-year-old woman was admitted to our hospital because of breathlessness. A diagnosis of advanced gastric cancer with extensive lymph node metastases and multiple pulmonary metastases was made. One hundred mg/body/day of TS-1 was orally administrated for 4 weeks. A partial response (PR) was obtained after the first course with regression of multiple pulmonary metastases. After 1 drug-free week, the second course was administered with 120 mg/body/day of TS-1 for 4 weeks. After two courses, the primary tumor was reduced to an ulcer scar with pathological confirmation of a complete disappearance of the cancer tissue. Moreover, computed tomography (CT) showed a complete regression of the extensive lymph node and diffuse lung metastases, for a complete response (CR). The serum level of CEA was reduced from 172.7 ng/ml to 8.1 ng/ml after TS-1 treatment. As for adverse events, only pigmentation of the skin and Grade 2 oral aphta were observed. PMID- 12090047 TI - [A case of unresectable gastric cancer complicated by a serious hepatic disorder and Virchow lymph node metastasis in which FP therapy and TS-1 administration were effective]. AB - The patient was a 65-year-old woman who came to our hospital because she had noticed swelling of a left supraclavicular lymph node. Endoscopy revealed an ulcerous lesion in the L chain region, and poorly differentiated adenocarcinoma of the stomach was diagnosed as a result of a biopsy. Oral therapy was impossible because of pyloric stenosis, and a gastrojejunal bypass operation was performed. Serious hepatic dysfunction occurred postoperatively due to swelling that was suspected of being due to lymph node metastasis in the porta hepatis. For this reason, the TS-1 therapy that had been initially scheduled was abandoned, and low dose FP therapy was instituted for 4 weeks. Afterward, the swelling resolved and the patient's hepatic disorder and systemic condition improved. Administration of TS-1, 80 mg/day (bid), was started in the outpatient clinic 4 weeks after the completion of FP therapy. Administration had to be discontinued after the first course of TS-1 therapy because of grade 3 thrombocytopenia and neutropenia. However, since marked decreases in tumor markers were observed, TS-1 administration was resumed after recovery. After completion of the second course of TS-1 therapy, a decrease in size of the Virchow lymph node was noted, and PR was diagnosed. TS-1 as initial chemotherapy had to be abandoned for this patient because of a serious hepatic disorder, in which TS-1 administration is contraindicated. However, a favorable response was obtained by concomitant use of FP therapy. PMID- 12090048 TI - [Successful treatment of gastric cancer accompanied by multiple liver metastases with TS-1 followed by curative gastrectomy affiliation]. AB - Despite the improvement in the operative results for patients with gastric cancer, the prognosis of those with liver metastasis remains dismal. Multimodal therapy has attracted considerable attention as a breakthrough in the strategy for treating cases of highly advanced gastric cancer. We report the case of a 62 year-old female with gastric cancer accompanied by multiple liver metastases successfully treated by TS-1, a novel oral fluoropyrimidine derivative. One treatment course consisted of 4 weeks of TS-1 administration (100 mg daily) followed by a 2-week break. After 3 courses of treatment, an abdominal CT scan showed no evidence of liver metastases and gastroscopy revealed that the primary gastric lesion was reduced. Grade 1 toxicity (nausea and diarrhea) was seen but 6 days rest improved the condition. Distal gastrectomy was subsequently performed without any finding of residual tumor. TS-1 may have a promising role in neoadjuvant chemotherapy for gastric cancer. PMID- 12090050 TI - [A case of advanced colon cancer with para-aortic lymph node metastasis responding greatly to treatment by adjuvant chemotherapy of low-dose CPT-11 on an outpatient basis]. AB - We administered an adjuvant chemotherapy of a bi-weekly low-dose CPT-11 + 5-FU on an outpatient basis to a patient with advanced sigmoid colon cancer and para aortic lymph node metastasis. After 3 months, tumor markers fell to normal levels and the regression rate in the lymph node was over 80%. Throughout the period of treatment, there were no side effects, and there was no sign of recurrence for 6 months. In conclusion, we consider that with low-dose CPT-11 + 5-FU chemotherapy on an outpatient basis, the patient can maintain a high quality of life and show a massive effect, and that this regimen can be used as a first-line chemotherapy for advanced colon cancer. PMID- 12090049 TI - [A case of malignant lymphoma of the stomach in which preoperative chemotherapy provided a complete response]. AB - We have encountered a case of malignant lymphoma of the stomach in which a complete remission was confirmed in a resected specimen after chemotherapy. A 75 year-old woman complained of vomiting blood. A biopsy from gastric endoscopy indicated malignant lymphoma of diffuse large B-cell type. The patient was assumed to be inoperable due to enlargement of the tumor and lymph node metastasis, and THP-COP chemotherapy was carried out. After four courses of the THP-COP regimen, endoscopic examination revealed a significant tumor reduction. Total gastrectomy and splenectomy with lymph node dissection (D2) were performed after chemotherapy. No tumor cells were detected in any sections of the specimen or regional lymph nodes. PMID- 12090051 TI - [A case of colon cancer with multiple liver metastases and lymph nodes metastases successfully treated by 5'-deoxy-5-fluorouridine]. AB - In a 79-year-old man with ascending colon cancer, multiple liver metastases were detected by computer tomography. The patient underwent right hemicolectomy. Paraaortic lymph nodes metastases were found intraoperatively. After surgery, 5' deoxy-5-fluorouridine (5'-DFUR) was administered orally at 600 mg daily. Abdominal CT showed disappearance of the liver metastases 6 months after operation. At a follow-up exam 24 months after the beginning of the chemotherapy, the patient remained in remission and reported no side effects. We consider oral administration of 5'-DFUR to be a useful treatment for advanced colon cancer. PMID- 12090052 TI - [Effectiveness of prolonged oral administration of low-dose etoposide in a case of malignant lymphoma in the mesenterium]. AB - Malignant lymphoma in the mesenterium is rare and carries a poor prognosis, although curative resection cases have been reported. A 39-year-old woman with non-resectable mesenteric malignant lymphoma obtained a better QOL and outcome from prolonged oral administration of low-dose etoposide as a maintenance therapy after CHOP therapy. PMID- 12090053 TI - [SNP project in the Millennium Genome Project, Japan]. AB - The basic outlines of the Millennium Genome Project were drawn by the Japanese government on December 19, 1999. In the "Disease gene" team, 5 disease have been chosen as the targets of the Project, Alzheimer's disease, cancer, diabetes, hypertension and asthma. Along with the knowledge-based candidate gene approaches, the complementary strategy of the disease gene hunting, the statistics-based approach, is being deployed. The Project decided to go for a genome-wide, gene-based SNP scan through a close collaboration with another team of the Project, "Standard polymorphism" team, which already discovered c. a. 160,000 SNPs in and around the genes of the Japanese people. PMID- 12090054 TI - [Hemochromatosis: from phenotype to genotype]. PMID- 12090055 TI - [Plasma homocysteine levels in Spanish patients with coronary artery disease]. AB - OBJECTIVE: A rise in plasma levels of the amino acid homocysteine (HCY) is a possible risk factor in cardiovascular disease. The mechanisms proposed to explain how HCY can increase the risk of vascular disease include its direct effect on the vascular endothelium and its role in increasing the risk of thrombosis. The present work has been designed to determine HCY levels in patients with coronary artery disease (CAD) residents in the Canary Islands and to establish whether hyperhomocysteinemia can be considered as an risk factor. METHODS: The sample studied consisted of 132 patients with, angiographically demonstrated, CAD and 18 controls with normal coronary arteries. Biochemical parameters determined included: HCY, vitamin B12, vitamin B6, folic acid, creatinine, cholesterol and its fractions, triglycerides, glucose and fibrinogen. RESULTS: Mean levels of HCY were not significantly different between the cases and controls (p = 0.37). In the distribution of HCY levels into quintiles there was no significant association between the quintiles and the occurrence of CAD (p = 0.57). Multiple logistic regression analysis in which the risk factors were compared with quintiles 2, 3, 4 and 5 of HCY did not reveal a significant relation between HCY levels and risk of CAD. CONCLUSIONS: This study questions the previously accepted consideration that hyperhomocysteinemia is a risk factor of CAD. Controlled intervention trials are, therefore, necessary to clarify the possible association between total HCY levels and cardiovascular disease. PMID- 12090056 TI - [Acute aortic dissection]. AB - OBJECTIVE: Acute aortic dissection is a medical emergency associated with high rates of mortality. Despite recent diagnostic advances, prompt and accurate diagnosis is difficult. We present a review of the a medical emergency service experience to diagnosis, factors associated to prompt diagnosis and inhospital evolution. MATERIAL AND METHOD: We studied 86 consecutive patients who had aortic dissections, to assess the presentation, management, and outcome. RESULTS: Mean age 61.9 +/- 12.5 years, 80% male. Hypertension was the most common predisposing factor (65% of patients overall). Chest pain was the most common initial complaint (58%). The initial clinical impression was aortic dissection in 56% of patients overall. Computed tomography was diagnostic in 96% when used. 62% underwent surgery. Overall in-hospital mortality was 28%. CONCLUSIONS: Acute aortic dissection presents with a wide range of manifestations. A high clinical index of suspicion and a prompt computed tomography should to improve recognition of aortic dissection. PMID- 12090057 TI - [Doxazosin, of modified liberation, in hemodialyzed patients]. AB - INTRODUCTION: Valuation of doxazosin, system formulation modified, in the hypertension in the hemodialysis population. METHOD: Arterial Hypertension (AHT) has been studied in 77 patients (p) subjected to hemodialysis (HD). Mean age (mag) was 61 years (y), range 84y-25y; 66% were males. The underlying etiology was glomerular in 19%, tubulo-interstitial in 18%, congenital in 18%, vascular in 19% and diabetic in 26%. RESULTS: Doxazosin (system formulation modified, single daily dose (4 mg), treatment follow-up was completed in 16 patients subjected to HD for 24 weeks (wk). CONCLUSION: It is concluded that AHT is of great importance in HD, and can be adequately controlled with the new antihypertensive drugs. In this context, doxazosin affords excellent therapeutic control, efficiency and good pharmacological tolerance. PMID- 12090058 TI - [Gastrointestinal disorders in Guatemala and their relation with parasitic infections]. AB - BACKGROUND: To determine the frequency of symptoms sugestive of functional gastrointestinal disorders and their relation to parasite infections in a developing nation. MATERIAL AND METHODS: 174 consecutive patients was seen in a gastroenterology clinic in Solola (Guatemala). RESULTS: Symptoms suggestive of irritable bowel syndrome was diagnosed in 37 patients (21%), irritable bowel syndrome with dyspeptic syndrome in 26 (15%) and dyspeptic syndrome in 24 (14%). Forty-two patients (24%) had been diagnosed of parasitation in the past. Such patients had a higher risk of suffering symptoms suggestives with functional gastrointestinal disorder (OR 6,335 CI 95% 2.72-14-75). Fifty-nine patients (34%) were subjected to parasite examination in stools; of these, 13 proved positive. In this same group of patients the risk of functional gastrointestinal problems was likewise higher among the subjects with parasite infection (p < 0.05). In turn, 41 patients (49%) with symptoms suggestives of gastrointestinal functional disorders were or had previously been diagnosed of parasite infection. CONCLUSIONS: Symptoms suggestives of functional gastrointestinal disorders are frequent in developing regions and its was in relation to intestinal parasites. PMID- 12090060 TI - [Mycobacterium kansasii lung infection and synchronous bronchopulmonary carcinoma]. AB - The association of lung tuberculosis and carcinoma of the lung is very well known, although it is exceptional the synchronous presentation of lung infection for nontuberculous mycobacteria and lung carcinoma. We present the case of a 41 year old male, smoker, with antecedents of lung tuberculosis in the adolescence and chronic liver disease for virus B and C that he presents fever, hoarseness and loss of 4 kg of weight of a month of evolution. The CT scan of the chest revealed a lung mass in left apex with pneumonitis, tumor invading chest wall and mediastinal lymph node enlargement. The biopsy of a supraclavicular node showed metastasis of squamous carcinoma, while in the sputum was isolated Mycobacterium kansasii sensitive to rifampin and ethambutol. HIV was negative. The possible mechanisms of this uncommon association are commented, next to a revision of the literature. PMID- 12090059 TI - [Histiocytic necrotizing lymphadenitis or Kikuchi's disease. CT radiologic features]. AB - Histiocytic necrotizing lymphadenitis, Kikuchi Fujimoto's disease (KFD) is characterised by fever and lymphadenopathy, usually large cervical, unilateral lymph nodes. Such clinical presentation demands a work-up to exclude serious medical conditions like malignancy and infections. Foci of necrosis with lymphocytic Histiocytic predominance in association with scarce polymorphonuclear cells on lymph node examination, confirm the diagnosis of KFD. Many different patterns of computed tomographic (CT) appearance of KFD have been reported. We describe the CT scan finding in two patients with this disease. All our cases showed, after two and three weeks of evolution respectively, enlarged lymph nodes with hypodense centres and peripheral ring enhancement. These radiological alterations correlated with the central lymph node necrosis found in the anatomopathological studies. In conclusion, KFD should be considered in patients with fever, cervical lymph node enlargement and CT scan showing hypointense centres and peripheral ring enhancement. PMID- 12090061 TI - [Eye toxicity of antimalarial agents]. AB - We relate the evolution of a patient with Systemic Lupus Erythematosus with mucocutaneous, hematological, joint and renal expression. The skin lesions were treated with Chloroquine and Hydroxychloroquine and severe ocular toxicity related with antimalarials agent was developed. We revised the antimalarials use in Rheumatology, their utility, doses, adverse events and risk factors as cumulate doses, ideal daily doses, renal function and ophthalmological reviews recommendations. PMID- 12090063 TI - [Genetics of iron overload]. PMID- 12090062 TI - [Legionnaire's disease with predominant liver involvement]. AB - Like other pneumonias due to atypical agents, pneumonia due to Legionela Pneumophila has no characteristic clinical facts, although fever and non productive cough are almost constant and diarrhea with changes in mental status are common. Hyponatremia and moderate transient hypertransaminasemia are common too. Severe systemic affectation after hematogenous dissemination similar to those described with typical bacterial pneumonias is a prominent difference with other atypical agents, with high mortality rates in the absence of appropriate treatment. Etiological diagnosis is very difficult and it is normally achieved late in the course of the infection. Because of diagnostic difficulties and potential mortality in predisposed patients, empirical antibiotherapy has been extensively recommended. We present a patient affected by critical community acquired pneumonia due to Legionela Pneumophila serogroup 1 with liver alteration as the main manifestation and good response to empirical antibiotherapy with claritromycine and rifampin. We recommended the empirical use of such therapy in those pneumonias without microbiological diagnosis and torpid evolution. PMID- 12090064 TI - [Practical management of pleural effusion]. AB - There are many different diseases that can be associated with pleural effusions. When a pleural effusion is discovered, two questions need to be answered: 1) is the effusion a transudate or is it an exudate?, and 2) if the effusion is an exudate, what is the disease responsible for its production?. Answers to these questions can be obtained in more than two-thirds of patients testing the pleural fluid by diagnostic thoracentesis. The remainder may require watchful waiting until resolution or further diagnostic procedures, either non-invasive (radiologic imaging) or invasive (bronchoscopy, pleural biopsy, thoracoscopy). PMID- 12090065 TI - [Zieve's syndrome]. PMID- 12090066 TI - [Acute renal failure caused by renal artery embolism]. PMID- 12090067 TI - [Multicentric involvement in aggressive classical Kaposi's sarcoma]. PMID- 12090068 TI - [Pulmonary nodule in cirrhotic patient]. PMID- 12090070 TI - [Long-course coxalgia]. PMID- 12090069 TI - [Spontaneous bacterial peritonitis caused by Listeria monocytogenes]. PMID- 12090071 TI - [Sympathetic paroxysmal episodes secondary to cerebral anoxia]. PMID- 12090072 TI - High-potency sodium fluoride: a literature review. AB - A considerable number of clinical studies are documented in the literature on oral products containing 5,000 ppm fluoride (F), mainly nonabrasive gels. Study results show that this level of fluoride provides marked caries protection. These high-potency gels are especially effective for irradiated head and neck cancer patients, who typically develop rampant caries. Radiation therapy causes a marked decrease in salivation (xerostomia), which predisposes the teeth to caries formation and results in devastating consequences. The currently recommended treatment for these patients involves a 2-step regimen using a 5,000 ppm F gel and a conventional 1,000 ppm F dentifrice. Unfortunately, this treatment usually results in noncompliance because two steps are inconvenient for most patients. A 5,000 ppm F dentifrice meets the oral-hygiene needs of these patients. It provides an effective level of fluoride and a convenient one-step application procedure that increases patient compliance. PMID- 12090073 TI - The mechanism of fluoride protection. AB - Since the 1940s, researchers have understood the positive effect of fluoride on anticaries activity. Several studies, beginning with the Cheektowaga study, showed that an increased number of concentrated fluoride applications also have a beneficial effect on anticaries activity. In fact, the continuous presence of high-fluoride concentrations has a more significant protective effect than enamel fluoride. A high-potency dentifrice is the ideal form with which to produce protective effects. This article describes how high-fluoride concentrations affect dental caries. PMID- 12090074 TI - The benefits of high-potency fluoride dentifrices. AB - National surveys show that the incidence of caries in children, adults, and seniors is still quite high, despite interpretations to the contrary. The dental profession and general public agree that the standard level of 1,000 ppm to 1,500 ppm fluoride (F) in most dentifrices reduces dental caries. Many studies using higher concentrations of fluoride in dentifrices and gels, 2,000 to 5,000 ppm F, have shown that caries prevention increases with fluoride concentration. PMID- 12090076 TI - Case reports. 1. Xerostomia. PMID- 12090075 TI - Clinical manifestations of head and neck irradiation. AB - Treatment of squamous cell carcinoma of the oral cavity, salivary gland tumors, and Hodgkin's disease may include radiation therapy to the head and neck region. This therapy has treatment-related sequelae to the oral cavity and can contribute to increased rates of dental caries in these patients. To prevent radiation induced caries, patients are instructed to use a high-potency fluoride application in addition to their standard dentifrice during radiation therapy and the postradiation phases of their treatment. Traditionally, patients are prescribed a 5,000-ppm fluoride (F) gel that is applied using mouthguards. However, due to oral postoperative conditions, many patients are not complaint with this treatment. In many cases, the availability of a fluoride product that is easier to use could result in successful patient compliance. PMID- 12090077 TI - Case reports. 2. Impaired salivary function. PMID- 12090078 TI - Case reports. 3. Xerostomia. PMID- 12090079 TI - Case reports. 4. Desquamative gingivitis. PMID- 12090080 TI - Safety and efficacy of simultaneous bilateral primary combined trabeculotomy trabeculectomy for developmental glaucoma. AB - PURPOSE: To establish the safety and efficacy of simultaneous bilateral primary combined trabeculotomy-trabeculectomy for developmental glaucoma. METHODS: We studied 109 consecutive patients who underwent planned simultaneous bilateral primary combined trabeculotomy-trabeculectomy for developmental glaucoma by a single surgeon from January 1990 through December 1999. The main outcome measures were postoperative intraocular pressure (IOP), corneal clarity and diameter, visual acuity, bleb characteristics, time of surgical failure and complications. Postoperative complications including endophthalmitis and anaesthetic morbidity and mortality were also analysed. RESULTS: The series consisted of 218 primary combined trabeculotomy-trabeculectomy surgeries during 109 anaesthesias. The mean follow-up period was 16.33 +/- 16.22 months. The IOP reduced from 26.4 +/- 5.9 mmHg to 13.5 +/- 4.5 mmHg, with a mean percentage reduction of 46.2 +/- 23.7 (P < 0.0001). The success (IOP < 16 mmHg) probabilities were 90.9%, 88.0% and 69.3% at first, second and third year respectively (Kaplan-Meier analysis). The success probability of 69.3% obtained at third year was maintained till 6 years of follow up. One hundred and sixty six (76.1%) eyes had significant corneal oedema. Postoperatively, the cornea cleared in 93 (57.8%) eyes. Clinically, well functioning blebs were present in 114 of 171 eyes (66.6%). Postoperatively, 18 (8.3%) eyes developed shallow anterior chamber and 6 (33.3%) of them required surgical reformation. There was no incidence of endophthalmitis or any other sight-threatening complication. Of the anesthetic complications, apnea occurred in 17 (15.6%) patients and all were successfully resuscitated. The most serious post-anaesthetic complication was cardio-pulmonary arrest that occurred 5 hours postoperatively following aspiration during feeding in one child; this child could not be resuscitated. Two children had delayed recovery (2 and 4 hours respectively). The child who had delayed recovery by 2 hours survived and has completed 3 years of follow-up while the other child expired 48 hours later. CONCLUSION: Simultaneous bilateral primary combined trabeculotomy-trabeculectomy is safe and effective for developmental glaucoma. It obviates the need for long second anaesthesia with its attendant risks. It offers several other benefits to the patients and families. PMID- 12090081 TI - Of genes and disease. PMID- 12090082 TI - Timolol maleate 0.5% versus timolol maleate in gel forming solution 0.5% (Timolol GFS) in open angle glaucoma in India. Preliminary safety and efficacy study. AB - PURPOSE: To compare the efficacy and safety profile of Timolol maleate 0.5% versus Timolol gel forming solution (GFS) 0.5% in open angle glaucoma in Indian eyes. METHODS: In a prospective crossover study 52 patients of open angle glaucoma, well controlled intraocular pressure (IOP) on 0.5% timolol maleate solution were switched over to timolol GFS once a day, after a washout period of one month. A diurnal IOP measurement was done after 6 weeks and compared with patients on timolol maleate 0.5% twice a day. In addition, side effects reported or observed were compared. RESULTS: Statistically significant difference was not observed in ocular hypotensive effect of the two treatment. The side-effects in both the treatment groups were similar except for higher incidence of blurring of vision in patients on timolol GFS. The compliance was better with timolol GFS, but was not statistically significant. CONCLUSION: The results of this study suggest that the more convenient 0.5% timolol in gel forming solution can be offered as an equally efficacious and well-tolerated alternative to twice daily 0.5% timolol solution in open angle glaucoma. PMID- 12090083 TI - Combined extracapsular cataract extraction with ahmed glaucoma valve implantation in phacomorphic glaucoma. AB - PURPOSE: To report a retrospective analysis of a combined procedure of extracapsular cataract extraction (ECCE) with heparin surface modified (HSM) posterior chamber intraocular lens (PCIOL) implantation along with primary Ahmed glaucoma valve (AGV) implantation in an attempt to optimize visual acuity gains and intraocular pressure (IOP) control in patients with phacomorphic glaucoma. METHODS: ECCE with HSM PC IOL and AGV implantation was performed through two separate incisions in 15 patients diagnosed with phacomorphic glaucoma. Postoperative improvement in visual acuity and IOP control were monitored. RESULTS: A steady control of IOP was maintained in all patients with minimum anti glaucoma medications. The average visual acuity was approximately 6/24 at 3 months. CONCLUSION: Superior preoperative IOP control and a shorter phacomorphic attack resulted in better postoperative vision. The successful maintenance of IOP within the desired range in this study suggests that the procedure should be performed under similar conditions. PMID- 12090084 TI - Effect of combination antiretroviral therapy on cytomegalovirus retinitis. AB - PURPOSE: To study the various changes in the course of cytomegalovirus (CMV) retinitis following combination antiretroviral treatment. METHODS: Combination antiretroviral treatment was given to 12 patients with active CMV retinitis following which all anti-CMV medications were discontinued once the CD4 cell counts were > 100/mm3 for 3 months. RESULTS: The median CD4 cell count increased from 36.5/mm3 (range, 3-74/mm3) at baseline to 175.5/mm3 (range, 97-410/mm3) at 3 months. No patient had reactivation of CMV retinitis or developed extraocular CMV infection during median follow-up of 16.7 months. In one patient with peripheral active CMV retinitis, the retinitis resolved completely and remained so throughout the follow-up period without specific anti-CMV treatment. Five (41.7%) patients had immune recovery vitritis. CONCLUSION: Patients receiving combination antiretroviral treatment following treatment for CMV retinitis have better control of CMV retinitis but immune recovery vitritis is a common sequelae. Reactivation of CMV retinitis is common in patients who discontinue combination antiretroviral treatment. PMID- 12090085 TI - Further investigations on the association of Mycobacterium tuberculosis with Eales' disease. AB - PURPOSE: To apply polymerase chain reaction (PCR) on vitreous fluid (VF) from Eales' disease to further confirm its association with Mycobacterium tuberculosis. METHODS: Sixty nine VF samples from 69 patients (24 Eales' disease and 45 Non-Eales' as controls) were processed by conventional methods for detection of mycobacteria. Polymerase chain reaction (PCR) specific for IS 6110 and nested PCR (nPCR) using primers coding for MPB 64 gene were applied on all 69 VF. PCR based dot-blot hybridisation was applied on the IS 6110 amplified products of n PCR-positive VFs. RESULTS: Conventional methods (direct smear and culture) did not detect mycobacteria in any of the 69 VF samples. Five (20.8%) of 24 VF from Eales' and 2 (4.2%) of 45 VF from control patients tested positive for M. tuberculosis DNA by nPCR. This difference was statistically significant (P < 0.05). All 69 VF were negative by PCR for IS 6110. Two VF of Eales' patients positive by nPCR were also positive by DNA probe dot-blot hybridisation for IS 6110. CONCLUSION: Detection of M. tuberculosis DNA by PCR in a significant number of VF of Eales' disease patients reemphasizes the association of this bacterium with Eales' disease. PMID- 12090086 TI - Development of an immunoanalytical method for the detection of beta- and gamma crystallins and anti-crystallin antibodies. A molecular biomarker for cataract. AB - PURPOSE: To develop and evaluate an immunoanalytical method for the detection of beta- and gamma-crystallins and anti-crystallin antibodies. MATERIALS AND METHODS: Beta and gamma-crystallins isolated from rat lens were used as immunogens to raise polyclonal antibodies in rabbits. Antibody capture assay and western blot analysis showed that the antibodies to beta- and gamma-crystallins were specific. An indirect competitive enzyme linked immunosorbent assay (ELISA) developed to quantitate beta- and gamma-crystallin showed an IC50 value of 70 ng and 65 ng, respectively, based on regression analysis. Spiking studies with purified beta-crystallin antibodies showed that 33 ng of the purified antibody gave an absorbance of 1.1 at 450 nm, indicating the sensitivity of the method. RESULTS: Antibodies to beta- and gamma-crystallins were not detected in serum samples of the cataractous CFY/NIN rats (used as an animal model for induction of experimental cataract by feeding high galactose diet). However, the cataractous rat serum samples effectively displaced beta- and gamma-crystallin antibodies, indicating that these crystallins leak during cataract formation. The concentration of beta- and gamma-crystallins in the rat serum, as analysed by indirect competitive ELISA, was found to be in the range of 17.6-81.6 micrograms/ml [corrected] and 12.4-19.6 micrograms/ml, respectively. CONCLUSIONS: The methodology developed in the present study may find application as a biochemical tool in molecular epidemiology of cataract. PMID- 12090087 TI - Bipseudophakia. Clinicopathological findings of a pseudophakic human globe obtained postmortem implanted with an anterior chamber and a posterior chamber intraocular lens. AB - We report the clinicopathological findings of a human pseudophakic globe obtained postmortem, containing both anterior and posterior chamber intraocular lens, a condition we termed "bipseudophakia". PMID- 12090088 TI - Genes and diabetic retinopathy. AB - Several recent studies have provided evidence that good diabetes control is important to prevent diabetic retinopathy. However, some groups of patients develop diabetic retinopathy despite good control and others escape retinopathy despite poor control. This suggests the role of genetic factors in susceptibility to retinopathy. This article reviews the role of genetic factors in determining diabetic retinopathy. PMID- 12090089 TI - Bilateral anterior ischaemic optic neuropathy in Takayasu arteritis. AB - This report describes a case of a young male with bilateral, asymmetrical AION. He was subsequently diagnosed with Takayasu's disease. PMID- 12090090 TI - Pupillary block glaucoma following implantation of a posterior chamber pseudophakos in the anterior chamber. AB - Pupillary block glaucoma is a common complication of cataract surgery, especially following anterior chamber intraocular lens implantation. We report a case of pupillary block glaucoma with a posterior chamber IOL that was implanted in the anterior chamber following a complicated extracapsular cataract extraction. The case was successfully managed by explantation of the posterior chamber lens, anterior vitrectomy, peripheral iridectomy and secondary anterior chamber intraocular lens implantation. The intraocular pressure was controlled with a single topical antiglaucoma medication. PMID- 12090091 TI - A ruptured retinal arterial macroaneurysm presenting as subhyaloid haemorrhage. AB - We report a case of ruptured retinal arterial macroaneurysm with overlying subhyaloid haemorrhage. Both subhyaloid haemorrhage and subsequently the macroaneurysm was treated with argon laser photocoagulation. PMID- 12090092 TI - Silicone sleeve of extrusion cannula as an intraocular foreign body. AB - An unusual intraocular foreign body--the silicone sleeve of a soft-tipped extrusion cannula left behind accidentally in the vitreous cavity following a vitreoretinal surgical procedure for complex retinal detachment--is reported. The silicone sleeve remained within the eye for a year without causing any problem. PMID- 12090093 TI - Oncocytoma of the caruncle. AB - A rare case of oncocytoma of the caruncle in a 75-year-old female is reported. Clinical differential diagnosis and histopathologic features are discussed. PMID- 12090094 TI - Training primary care physicians in community eye health. Experiences from India. AB - This paper describes the impact of training on primary-care physicians in community eye health through a series of workshops. 865 trainees completed three evaluation formats anonymously. The questions tested knowledge on magnitude of blindness, the most common causes of blindness, and district level functioning of the National Programme for Control of Blindness (NPCB). Knowledge of the trainers significantly improved immediately after the course (chi 2 300.16; p < 0.00001). This was independent of the timing of workshops and number of trainees per batch. Presentation, content and relevance to job responsibilities were most appreciated. There is immense value addition from training primary-care physicians in community eye health. Despite a long series of training sessions, trainer fatigue was minimal; therefore, such capsules can be replicated with great success. PMID- 12090095 TI - Comparison of external dacryocystorhinostomy and 5-fluorouracil augmented endonasal laser dacryocystorhinostomy. A retrospective review. PMID- 12090096 TI - Efficacy of topical and systemic itraconazole as a broad-spectrum antifungal agent in mycotic corneal ulcer. A preliminary study. PMID- 12090097 TI - Efficacy of topical and systemic itraconazole as a broad-spectrum antifungal agent in mycotic corneal ulcer. A preliminary study. PMID- 12090098 TI - Efficacy of topical and systemic itraconazole as a broad-spectrum antifungal agent in mycotic corneal ulcer. A preliminary study. PMID- 12090099 TI - [Changes in the epidemiology of infectious diseases in Chile from 1990 to 2000]. AB - In the last decade in Chile, there was a large reduction in the rate of communicable diseases, especially typhoid fever, and those preventable through the universal vaccination programs. Exceptions were hepatitis A and Pertussis. The reduction in tuberculosis, has lead the country to a threshold in which the elimination of tuberculosis as a public health problem is reachable. The HIV epidemic is still expanding, at higher rate among women and heterosexual men, keeping drug addiction as a low ranked risk factor. At the end of the century, universal or expanded access to HIV therapy was still not a reality. Cholera was a well controlled emerging infection, but Hantavirus infection has become a major threat in many regions. Syphilis and especially gonorrhea have decreased, but condyloma has increased dramatically. The nosocomial infection scenario has changed somehow, due to more severely ice and complex patients admitted to hospitals. Multiresistant nosocomial pathogens continue expanding (S aureus, fermentor and non fermentor gram negative rods, especially A baumannii). The country has been able to expand control programs to almost all hospitals. Antimicrobial resistance has continued growing. The massive and indiscriminate use of antibiotics, largely responsible for the resistance, grew worse until the sale of antimicrobials in pharmacies was restricted by law. This had a major impact, with important reduction in sales of most, but not all, antibiotics. The impact in resistance rate of this reduction, if any, has yet to be assessed. PMID- 12090100 TI - [Immunophenotyping of peripheral blood lymphocytes from Chilean chronic chagasic patients by flow cytometry]. AB - BACKGROUND: Cellular immune mechanisms of the resistance to infection by T cruzi as well as the pathogenesis of Chagas disease are still controversial. AIM: To quantify and analyse the peripheral blood immune cells from chagasic and non chagasic patients by flow cytometry. PATIENTS AND METHODS: Peripheral blood samples were taken from 21 individuals seropositive for Chagas disease, under no specific treatment. Control samples from 21 healthy blood donors were also obtained. To quantify immune cells populations by flow cytometry, antibodies against CD3, CD4, CD8, CD16/56, CD45/14, CD19 and HLA-DR markers were used. RESULTS: The percentage of CD8+ cells was low and the CD4+/CD8+ ratio was high in chagasic patients, compared to their non infected counterparts. No statistically significant differences in the number of CD4+, NK, B, CD4+HLADR+ and CD8+HLADR+ cells, were observed within the two groups. CONCLUSIONS: These results show that Chilean chronic chagasic patients have lower percentage of CD8+ cells and higher CD4+/CD8+ ratio than non infected individuals. PMID- 12090101 TI - [Reduction of myocardial infarction mortality in Chilean hospitals]. AB - BACKGROUND: The characteristics of patients with acute myocardial infarction (MI) admitted to 37 Chilean hospitals (GEMI Registry Group), have been analyzed in the period 1993-1995 and 1997-1998. AIM: To report the changes in hospital mortality between these 2 periods, with a particular emphasis on the impact of treatment. PATIENTS AND METHODS: Between 1993-1995 we collected information from 2,957 patients and between 1997-1998 we registered 1,981 patients with MI. Analysis of the changes in mortality between periods was adjusted by demographic variables, coronary risk factors, MI location, Killip class on admission and the different therapeutic strategies utilized. The effects of different treatments on hospital mortality were adjusted by the previously determined mortality risk variables. RESULTS: Hospital mortality decreased from 13.3% to 10.8% between both periods (Odds Ratio (OR) 0.78, confidence intervals (95%) (CI) 0.65-0.93). A significant reduction in mortality was observed among patients below 60 years of age, in men, in diabetics and in subjects with an infarction classified as Killip class over II. The use of beta blockers (OR 0.65, CI 0.42-0.99) and intravenous nitrates (OR 0.78, CI 0.61-0.99) and the lower use of calcium channel blockers (OR 0.72, CI 0.60-0.87) were significantly associated with a lower mortality. The administration of angiotensin converting enzyme inhibitors was associated with a 29.3% mortality reduction (OR 0.69, CI 0.47-1.02). CONCLUSIONS: There has been a significant reduction in the mortality rate for MI in Chilean hospitals during the 2 registry periods analyzed, which was significant among some high risk patients and was related to treatment changes, according to evidence based guidelines. PMID- 12090102 TI - [Morbidity and mortality of treated essential arterial hypertension in a 26 years follow up]. AB - BACKGROUND: Treatment has a definitive impact on mortality in hypertension. The magnitude of blood pressure reduction, the type of drug used and the associated risk factors may modulate the effect of treatment on mortality. AIM: To report the effects of treatment of essential hypertension, in a cohort followed for up to 26 years. PATIENTS AND METHODS: A cohort of 1,172 essential hypertensive patients followed up to 26 years. Patients were treated with different antihypertensive drugs, alone or in combination (diuretics, beta blockers, calcium channel blockers and angiotensin converting enzyme inhibitors) according to international rules and consensus. Subjects were followed until death or loss from follow-up. Blood pressure reduction was aimed to obtain figures near 140/90 mm Hg. Causes of death and complications were obtained from hospital records, phone and death certificates. Survival was studied using life tables (Kaplan Meier method and intention to treat analysis) and Cox proportional hazard analysis. RESULTS: Initial blood pressure dropped significantly from 181/109 to 154/92 mm Hg, p < 0.001. Mean follow-up time was 10.6 +/- 6.1 years. There were 143 cardiovascular deaths, 142 acute myocardial infarctions, 101 strokes, 83 subjects had cardiac failure and 49 had renal failure. The observed rates were 0.92% per year for cardiovascular mortality, 1.36% per year for coronary heart disease, 0.94% per year for stroke. CONCLUSIONS: Our mortality rate is lower than that found in classical randomized studies but similar or slightly higher than the more recent ones. Morbidity rates were also very similar. Except for mortality rate, frequency of complications did not change in comparison to our previous report after 15 year of follow up. Aging did not seem to negatively influence mortality rates in adequately treated hypertensive patients. PMID- 12090103 TI - [Morphological prognostic elements in gallbladder cancer]. AB - BACKGROUND: The exact survival rates and prognostic factors of gallbladder cancer are still incompletely known. AIM: To report the actuarial survival of patients with gallbladder cancer. MATERIAL AND METHODS: Six hundred thirty seven women, aged 59 years old as a mean and 108 men, aged 64 years old as a mean, with gallbladder cancer are reported. Patients were followed for up to 150 months. RESULTS: Two hundred twenty four patients had an early and 521 had an advanced carcinoma. Overall survival was 38% at ten years. Sex or ethnic origin did not influence survival. Early tumors had a 92% survival at 10 years whereas the survival of advanced tumors was 16% at 5 years. Subserous tumors had a 5 years survival of 32% whereas serous tumors had a 5 years survival of 11%. Well differentiated advanced tumors had a significantly better survival than moderately or poorly differentiated tumors. Vascular or lymphatic infiltration was also associated to a lower survival. All patients with advanced tumors and vascular infiltration died before 5 years. CONCLUSIONS: Tumor infiltration and differentiation degree were the most important prognostic independent factors in gallbladder cancer. PMID- 12090104 TI - [Acanthamoeba keratitis treated with propamidine and polyhexamethyl biguanide (PHMB)]. AB - BACKGROUND: The diagnosis of acanthamoeba keratitis has increased since 1985 due to the massive use of contact lenses and a better knowledge of the disease by ophthalmologists. The use of biassociated therapy has resulted in a better prognosis and lower complication rate. AIM: To report patients with acanthamoeba keratitis treated with the association of propamidine (Brolene) and polyhexamethylbiguanide (PHMB) 0.02%. PATIENTS AND METHODS: Retrospective analysis of 27 patients (31 eyes) with acanthamoeba keratitis (bilateral in four cases), diagnosed by culture, biopsy or characteristic clinical features. RESULTS: Ninety six percent of patients used rigid contact lenses. Acanthamoeba cultures were positive in 71% of cases. The delay in the diagnosis was between 1 and 5 months. Early treatment was possible in 29% of patients. Infection was irradicated in all cases with the biassociated therapy. A tectonic keratoplasty to treat a trophic perforation was done in eight eyes. No patient required therapeutic keratoplasty to resolve the infection. Visual acuity at the end of follow up was better than 20/40 in nine patients and in eight it was in the range of count fingers or less. CONCLUSIONS: In patients with the clinical picture of acanthamoeba keratitis, early or late antiamoebic treatment is warranted even in the absence of positive cultures. The visual results of the treatment are highly dependent on the precocity of treatment. Prevention is imperative and is based on a strict contact lens hygiene. PMID- 12090106 TI - [Study of the chemical stability of acetylsalicylic acid tablets during storage in pharmacies of Concepcion, Chile]. AB - BACKGROUND: The chemical stability of a pharmaceutical product depends, among other factors, on environmental factors during transport, storage and manipulation of the product. AIM: To study the chemical stability of acetylsalicylic acid (AAS) tablets during ten months of storage in five pharmacies of Concepcion, Chile. MATERIAL AND METHODS: Tablets were randomly collected at the beginning of the study and at the third, sixth and tenth month. Quantitative analyses of AAS tablets was carried out by instrumental thin layer chromatography (HPTLC). RESULTS: AAS in tablets was between 99 and 109% at the beginning of the study, between 76 and 110% at three months, between 71% and 112% at six months and between 86 and 110% at ten months of storage. CONCLUSIONS: There was a progressive decrease in the content of acetylsalicylic acid in tables during storage, but it remained between the limits accepted by the United States Pharmacopoeia (USP) (90-110%). PMID- 12090105 TI - [Periodontal health status in patients with diabetes mellitus type 1, from 18 to 30 years-old, from Santiago de Chile]. AB - BACKGROUND: The prevalence of periodontal diseases, gingivitis and periodontitis, is higher in diabetic patients and can have severe functional and esthetic consequences early in their lives. AIM: To evaluate the prevalence of periodontal disease in type 1 diabetics, aged between 18 and 30 years old, living in Santiago de Chile. SUBJECTS AND METHODS: One hundred male and female type 1 diabetics were examined. Glycated hemoglobin A1c, microalbuminuria, and fundoscopy were assessed in a sample of 52 subjects, separated in two groups according to the presence of periodontal disease. RESULTS: The prevalence of gingivitis was 22%, periodontitis 41%. Only 37% of subjects were free of periodontal disease. When compared with patients without periodontal disease, in the group of patients with the disease there was a higher proportion of subjects with diabetes lasting more than 10 years (28 and 55% respectively) and a higher proportion of patients with chronic complications of diabetes (42 and 58% respectively). CONCLUSIONS: A high prevalence of periodontal diseases was observed in this sample of diabetic patients. A long history of diabetes and the presence of chronic complications were risk factors for these diseases in the analyzed sample. PMID- 12090107 TI - [Identification and antifungal susceptibility of Candida spp isolated from invasive mycoses. Influence of growth inhibition percentage to determine minimal inhibitory concentration]. AB - BACKGROUND: An increase in the frequency of resistant strains to antifungal drugs has been detected in the last decade. AIM: To report the minimal inhibitory concentration (MIC) to amphotericin B, fluconazole, ketoconazole and itraconazole. To compare the MIC obtained with 80% and 50% of growth inhibition to the azoles. MATERIAL AND METHODS: Fifty yeast strains isolated between 1998 and 1999, from 17 adults and 33 children with invasive mycosis were studied. Susceptibility was determined by broth microdilution method with RPMI 1640 plus glucose 2% according to the National Committee for Clinical Laboratory Standards (1997). RESULTS: The most frequently isolated strains were C albicans in 27 cases and C parapsilosis in 12. All isolates were susceptible to amphotericin B. According to the MICs obtained with 80% of inhibition, 12 strains had MICs considered as resistant to azoles. Five strains were resistant both to fluconazole and itraconazole. Considering MICs obtained with 50% of inhibition, only five strains were found resistant to azoles (p < 0.05). Using this criterion, only one C glabrata strain was found to be simultaneously resistant to fluconazole and itraconazole. CONCLUSIONS: Similar results in the pattern of susceptibility of Candida spp to azoles, to those reported abroad, are obtained when the MIC is calculated using 50% inhibition. PMID- 12090109 TI - [Irreversible acute heart failure caused by cardiac hemochromatosis secondary to multiple transfusions]. AB - We report a 64 years old male, admitted for dyspnea and edema of two weeks duration. A sideroblastic anemia, requiring multiple transfusions, was diagnosed four years earlier. The chest X-ray showed an enlarged heart and right pleural effusion. A low left ejection fraction was evidenced by echocardiogram. Doppler analysis of the mitral flow revealed a restrictive hemodynamic pattern. A diagnosis of secondary cardiac hemochromatosis deposit was made. Nine days after admission the patient died due to heart failure. The clinical presentation of cardiac hemochromatosis as a sudden and irreversible heart failure, as well as the importance of early diagnosis and surveillance of high-risk patients is emphasized. PMID- 12090108 TI - [Idiopathic endometrial decidual reaction in a woman in postmenopause: immunocytochemical evaluation of estrogen and progesterone receptors and its corticoid binding globulin]. AB - Endometrial decidual reaction in postmenopausal women without an evident exogenous or endogenous progesterone stimulus, is uncommon. The morphological findings are decidual transformation of the endometrial stroma, necrosis, cellular pleomorphism and in some cases hyperplastic glands that exhibit focal atypia raising the possibility of malignancy. We report the case of a 55 years old woman with an idiopathic endometrial decidual reaction. In this woman, immunocytochemistry of endometrial tissue demonstrated the absence of cytoplasmatic and nuclear estrogen receptors. Staining of nuclear progesterone receptors and cytoplasmatic corticoid binding globulin, exceeded the immunoreactivity observed in any period of the menstrual cycle. PMID- 12090110 TI - [Spontaneous hemoperitoneum caused by intraabdominal variceal rupture in a patient with liver cirrhosis. Clinical case]. AB - Spontaneous hemoperitoneum secondary to intraabdominal variceal rupture, is an uncommon and highly lethal complication of cirrhosis. We report a case of a 68 years old male submitted because of abdominal pain and fainting. The clinical and laboratory work up concluded that the patient had cirrhosis and hemoperitoneum. He was operated on and the exploration showed a massive hemoperitoneum caused by bleeding gastrosplenic varices. The patient died in the immediate postoperative period. PMID- 12090111 TI - [Improvement of medical student's academic performances in times of curricular reform]. AB - BACKGROUND: During the last decade, academic life at the medical school of the Pontificia Universidad Catolica de Chile has been thoroughly affected by a curricular reform process. Changes started in 1993 and have continued up until now. This reform did not have an experimental design to allow for a scientific evaluation of its effects. However, it seems interesting to study the evolution of indices of academic performance of our students during this period. AIM: To evaluate the academic performance of medical students between 1989 and 1999. SUBJECTS AND METHODS: All undergraduate students enrolled between 1989 and 1999. Academic performance was evaluated by 1) failure to pass one or more courses, 2) delay to complete the third year of studies, 3) withdrawal from school and 4) medical school grades. RESULTS: All indices of academic performance changed during the period of study. Failure to pass, delay and withdrawal from school significantly decreased, whereas medical school grades improved, particularly in basic and pre-clinical subjects. CONCLUSIONS: Academic performance of medical students improved consistently between 1989 and 1999. While specific causal relationships cannot be established, we believe that this improvement is likely related to the curricular reform. This reform included horizontal and vertical integration of academic contents, greater emphasis in problem-based learning and additional instances of evaluation such as the repetition exam. PMID- 12090112 TI - [Cost-effectiveness analysis of a preventive program for gallbladder disease in Chile]. AB - BACKGROUND: Chilean women have one of the highest mortality rates from gallstone disease in the world. There is no primary prevention for the disease and the benefits of prophylactic cholecystectomy in high risk groups have not been studied. AIM: To analyze the cost and effectiveness of a screening program for gallbladder disease in the Chilean women population. METHODS: A decision analytic model is used to compare lifetime cost and effectiveness of standard care with three screening strategies. The first two strategies consider "universal ultrasound screening" for all women 40 years old and laparoscopic cholecystectomy for those with gallstones ("elective intervention") or with calculous > or = 3 cm ("high risk intervention"). The third strategy is based on "selective screening" for obese women. RESULTS: The lifetime probability of a 40 years old Chilean woman of dying from gallbladder disease is reduced by 70% in the universal screening/elective intervention, by 63% in the high risk intervention and by 18% in the selective screening strategy. Her lifetime expectancy increases by 5.25, 4.64 and 1.24 months respectively. The incremental cost-effectiveness ratio of each screening strategy is US$ 180, US$ 147 and US$ 481 respectively. CONCLUSION: A screening program for gallbladder disease in a high risk population achieves significant benefits at a low incremental cost and acceptable cost-effectiveness. PMID- 12090113 TI - [Chromosome 8 trisomy in 2 cases of negative ABL/BCR chronic myeloid leukemia ]. PMID- 12090114 TI - [Assessment of clinical competence in medical education: a need]. PMID- 12090115 TI - [Psychology of hair in the past and today. From roundheads and wigs, from skinheads and hair styling]. PMID- 12090117 TI - [Diffuse hair loss in women]. AB - The complaint "Doctor, I am losing my hair" represents a particular challenge to the physician, and involves making a specific diagnosis, selecting an appropriate therapy, and expressing empathy for the patient's anxiety. Diffuse hair loss in women was formerly classified as an entity of its own. Since the identification of female pattern hair loss, most cases have been recognized to be due to androgenetic alopecia, often during phases of life characterized by fluctuations of sexual hormone levels or in connection with intake or cessation of hormonal therapy. The most difficult differential diagnosis includes androgenetic alopecia, chronic telogen effluvium, and psychogenic pseudo efflvuium. Androgenetic alopecia is due to androgen-induced, non-synchronized, progressive shortening of the hair growth cycle and gradually leads to thinning of the central scalp area. Idiopathic chronic telogen effluvium typically occurs in women, starting abruptly without a recognizable initiating factor, and involves the entire scalp area with increased shedding of telogen hair. It is believed to be due to synchronization phenomena of the cyclic hair growth. Psychogenic pseudo effluvium affects fashion-oriented, self-conscious women suffering of a discrepancy between the actual state of their hair and idealized expectations. Later the problem of age-related hair thinning oft becomes a surrogate for the more generalized problem of senescence. Rational therapy of androgenetic alopecia aims at blocking the androgen effect on hair follicles with estrogens and antiandrogens or at pharmacologically reversing vellus hair transformation with topical minoxidil. In contrast, women with idiopathic chronic telogen effluvium should be reassured that their problem is rather a state of exaggerated "hair shedding" than of actual "hair loss". PMID- 12090116 TI - [Androgenetic alopecia in the man]. AB - Androgenetic alopecia (AGA) occurs in approximately 40% of men at the age of 40 and 50% at 50, respectively. Especially for young men progressive hair loss can be distressing. Therefore, understanding of these patients' concerns is important for appropriate management. Current understanding of the pathophysiology of AGA mainly focuses on androgen metabolism as it affects hair growth. As a result, pharmacologic treatment has made considerable progress through the introduction of selective 5 alpha-reductase inhibition with finasteride. In placebo-controlled clinical trials in men with AGA, treatment with oral finasteride proved to be effective. Minoxidil is the only pharmacological substance for topical application with proven efficacy. So far, other treatment modalities have no proven efficacy in clinical trials, so that their use cannot be recommended. Options for advanced AGA not amenable to pharmacologic treatment are autologous hair transplantation and hair replacement, both of which have recently also made progress in terms of cosmetic appeal. PMID- 12090118 TI - [Hair diseases in childhood]. AB - This paper focuses on four important hair diseases mainly occurring in children. Trichotillomania is the most relevant differential diagnosis of alopecia areata in childhood. Meticulous inspection and lack of telogen hairs in the trichogram from the margin of the lesion usually are sufficient for differentiation. The trichogram also plays a significant role for the diagnosis of the loose anagen hair (loose anagen syndrome), a fairly new, but not rare entity, especially in distinguishing it from telogen effluvium. Five different types of clinical presentation are distinguished in tinea capitis. For the necessary systemic therapy; the new antimycotics terbinafine and itraconazole represent good alternatives to the well-tried griseofulvin. Several effective therapeutic options are also available for head lice, the most frequent parasitary infestation of school age. However, because of its neurotoxicity and the increasing problem of lice resistance lindane should not be used any longer for the treatment of head lice. PMID- 12090119 TI - [Hair shaft abnormalities]. AB - Hair shaft disorders may lead to brittleness and uncombable hair. In general the hair feels dry and lusterless. Hair shaft abnormalities may occur as localized or generalized disorders. Genetic predisposition or exogenous factors are able to produce and maintain hair shaft abnormalities. In addition to an extensive history and physical examination the most important diagnostic examination to analyze a hair shaft problem is light microscopy. Therapy of hair shaft disorders should focus to the cause. In addition, minimizing traumatic influences to hair shafts, such as dry hair with an electric dryer, permanent waves and dyes is important. A short hair style is more suitable for such patients with hair shaft disorders. PMID- 12090120 TI - [Alopecia areata]. AB - Alopecia areata is a frequent cause of hair loss. The origin of disease is not fully understood. However there are indications for a T-cell mediated autoimmune process. Genetic, immunologic and psychologic factors are important for the outbreak of disease. Most patients show localized patches of acute hair loss, where regrowth is observed spontaneously or with simple topical treatment within few months. In up to 15% of patients severe forms of disease can develop with total scalp (alopecia totalis) or scalp and body hair loss (alopecia universalis). There are only few known risk factors for development of a severe form. Although spontaneous remission is possible in these cases, it occurs rarely and treatment is difficult. Multifocal alopecia areata responds to intravenous high-dose corticosteroids. Topical immunotherapy with diphenylcyclopropenone (DPC) or PUVA therapy may be effective in longstanding and widespread disease. The unpredictable course of disease is a major handicap for clinical trials and treatment recommendations. Contact of patients with self-help organisations may be of help for coping with the disease. PMID- 12090121 TI - [Hair loss in internal medical illnesses]. AB - Hair loss related to internal diseases is generally temporary and often fully reversible. An iron- or protein-deficiency induced hair loss may be cured by simple substitution. In acute internal diseases, fever and after operations the patient may expect complete recovery of the hair loss without therapy. Symptomatic alopecia due to chronic diseases has a different prognosis and is dependent on the severity and character of the underlaying disease. If the systemic disease can be cured the hair loss may be decreased. Treatment and diagnosis of the systemic disease is recommended to be performed in cooperation with experts of internal medicine, oncologists and specialists of endocrinology. PMID- 12090122 TI - [Scarring alopecias]. AB - The irreversibility and the possible important cosmetic consequences of scarring alopecia demand special diagnostic attention in order to promptly attain a precise diagnosis and specific treatment. Scarring alopecias are either due to permanent damage to essential parts of the hair follicle or destruction of the entire hair follicle. They are classified into the categories of primary scarring alopecias, where the hair follicle is the primary target of destruction, and secondary scarring alopecias, where the follicular damage results incidentally from events around impinging on the follicular unit. The differential diagnosis of the more common primary scarring alopecias, e.g. follicular lichen planus, chronic cutaneous lupus erythematosus and folliculitis decalvans, can be difficult when based only on anamnestic and clinical findings. The scalp biopsy is essential for appropriate nosologic classification and has prognostic relevance. The primary therapeutic goal is to halt progression of the irreversible process as early as possible by means of immunomodulatory, immunosuppressive or antiinfectious agents, respectively. PMID- 12090123 TI - [Hypertrichosis and hirsutism]. AB - Facial hypertrichosis and hirsutism may cause severe cosmetic and psychologic problems. There are several therapeutic options, both on the cosmetic and medical level. Hirsutism with androgen-excess should be diagnosed and treated by gynecologists. Although frequently used, systemic antiandrogens have not yet proven their efficacy against unwanted hypertrichosis in clinical studies using modern and objective endpoints. Conventional methods of hair removal include plucking, waxing and chemical depilation by thiogycolates, as well as electrolysis and thermolysis. A new medical treatment of hypertrichosis is eflornithine cream. It inhibits the enzyme ornithine-decarboxylase which is essential for the rapidly dividing cells of the hair follicle. Permanent depilation by photothermolysis with lasers and pulsed flash-light systems is currently the most promising treatment. PMID- 12090124 TI - [Shampoo]. AB - The diversity of qualities demanded from a shampoo by today's consumer go beyond the sole function of cleansing the hair and scalp. A cosmetic benefit is expected, and the shampoo formulation has to be tailored to all the possible variations associated with hair quality (dry, greasy, permed, bleached, dyed), age (baby shampoo), care habit (frequency of shampooing) and specific problems relating to the superficial condition of the scalp (dandruff, seborrhea). The standard graduate medical training programs devote limited attention to the subject of shampoos, and much of the readily available information concerning shampoos is supplied by the industry. We should be increasingly aware that our patients look to us to supply information on what can be expected from a shampoo to enable them to make more informed choices at the consumer level. Moreover, selected ingredients of shampoos that have been popular with the consumer are currently under attack because of potential risks associated with their use, e.g. halogenated organic compounds, formaldehyde, nitromusc, and crude coal tar. Their critical appraisal also has to follow strict scientific criteria. PMID- 12090126 TI - Cardiopulmonary exercise test--its contribution in cardiology, among other things. PMID- 12090125 TI - Recovery kinetics of oxygen uptake after cardiopulmonary exercise test and prognosis in patients with left ventricular dysfunction. AB - INTRODUCTION: The prognostic value of peak oxygen uptake (peak VO2) in patients with left ventricular systolic dysfunction is currently recognized and accepted. Some studies have shown that other cardiopulmonary exercise test (CPET) parameters have additional value. OBJECTIVES: To evaluate whether our population of patients with left ventricular dysfunction had similar results to those found by other investigators who showed that a slow normalization of oxygen uptake (VO2) during the recovery period of a CPET has prognostic value, and whether the recovery phase parameters have additional prognostic value to peak VO2 in these patients. METHODS: We studied 292 consecutive adult patients (81.5% male; mean age 52.4 +/- 10.6 years) with an ejection fraction below 40% (mean 23.6 +/- 8.8%) given their first symptom-limited CPET between 03/1993 and 08/2000. The etiology was ischemic heart disease in 154, idiopathic cardiomyopathy in 130 and other in 8 patients. NYHA class was I in 7%, II in 50.6% and III in 42.4% of the patients. Two years was defined as the maximum follow-up time; it was 551.5 +/- 242.2 days on average, and 62 events (death or cardiac transplantation) occurred. The following parameters were analyzed: peak VO2 (l/min and ml/kg/min), percent predicted peak VO2 (pred VO2) (l/min and ml/kg/min), VO2 every 15 seconds (sec) of the first 3 minutes of recovery (the difference between peak VO2/kg and VO2/kg every 15 sec in the recovery period (dif VO2), expressed in ml/kg/min, and also the time (sec) to reach 50% of peak VO2 (T1/2). It was considered that a combined end-point was reached if patients died or underwent cardiac transplantation. RESULTS: ROC curves of these parameters showed the following as cut-off values (area under the curve > 0.7) for the occurrence of events: peak VO2 < 60% of pred VO2, dif VO2 at 60 sec (< 3 ml/kg/min), 90 sec (< 5), 120 sec (< 8), 150 sec (< 8.6) and 180 sec (< 10.5) of the recovery and T1/2 > 115 sec. Survival analysis was performed considering pred VO2 < 60%, dif VO2 at 150 sec (the largest area under the curve) and T1/2 > 115 sec. In the survival analysis, when the decrease in VO2 at 150 sec was less than 8.6 ml/kg/min the number of patients with events increased from 9.2% to 43.5% (p < 0.001; log-rank), and when T1/2 was less than 115 sec the number of events increased from 12.3 to 34.2% (p < 0.001; log-rank). When the criteria of T1/2 and dif VO2 at 150 sec were considered together with pred VO2 < 60%, mortality increased from 31 to 54% and from 33 to 51%, respectively (p < 0.001, for both parameters; chi-square). CONCLUSIONS: A slow VO2 kinetics in the recovery period of the CPET by itself identified groups of patients with poor prognosis. The association of these parameters with peak VO2 enhanced the identification of groups at greater risk for events. A global evaluation of the CPET should be performed, considering other parameters besides peak VO2, particularly those related to VO2 kinetics in recovery (T1/2 and dif VO2 at 150 sec) as identified in this study. PMID- 12090127 TI - Electronanatomical mapping and ablation of atrial tachycardias with the CARTO system. AB - Variable success rates in the ablation of atrial tachycardias using conventional electrophysiology have been achieved. There is no precise relation between P wave morphology in surface ECG and atrial electrophysiology, and this fact makes it more difficult to locate ectopic atrial foci. The CARTO system creates atrial activation maps that relate an anatomical location to an electrical potential. The aim of this study was to evaluate the efficacy of CARTO guided radiofrequency (RF) ablation of atrial foci. The population consisted of 10 consecutive patients with atrial tachycardia resistant to more than 2 drugs, 7 female, mean age 45 +/- 12 years. CARTO activation maps were constructed based on atrial tachycardia or premature beats. Radiofrequency energy was applied to the earliest activation zone. Immediate success was defined as suppression of ectopic atrial activity. Ectopic foci were located on the ostium of the coronary sinus (3 patients), crista terminalis (1 patient), right atrial appendage (1 patient), interatrial septum (1 patient) and in the pulmonary veins (4 patients). The activation maps contained 85 +/- 35 points. The number of RF applications ranged from 1 to 11 (mean 4). Immediate and 6 month success rate was 90%. We were not able to treat one patient with a focus in the right atrial appendage. No attempt was made to limit procedure or fluoroscopy time in our study. Nonetheless all procedures lasted less than 150 min, and fluoroscopy times were less than 30 minutes. CONCLUSIONS: The CARTO system precisely located ectopic atrial foci, allowing a high success rate in the ablation of focal atrial tachycardias. PMID- 12090128 TI - Valve orifice area in aortic stenosis evaluated by planimetry, Gorlin and continuity equations: a prospective study. AB - In evaluation of the severity of aortic valve stenosis, multiple parameters can be determined. All of them, except valve orifice area, are influenced by other factors such as cardiac output, heart rate or aortic insufficiency. OBJECTIVES: This is a prospective study which proposes, in the determination of the valve orifice area in aortic stenosis, to evaluate the accuracy of and correlation between three methods--planimetry by multiplane transesophageal echocardiography, the continuity equation by transthoracic echocardiography, and invasive measurement using the Gorlin formula. METHODS: Forty-five patients with known calcified valvular aortic stenosis 27 men, mean age 70 +/- 10 years, (range 27 82), were studied. In all patients the area was determined by planimetry and by the continuity equation. In 25 (56%) patients invasive measurements were obtained using the Gorlin formula. RESULTS: Evaluation of the valve orifice area by planimetry was easily performed and did not prolong the duration of the exam, except in five patients (11%). The area determined by the continuity equation had a mean value of 0.74 +/- 0.25 cm2, by planimetry 0.74 +/- 0.24 cm2 and by the Gorlin formula 0.65 +/- 0.17 cm2. Correlations between areas obtained by the three methods used were: continuity equation and planimetry 0.82; continuity equation and Gorlin formula 0.51; and planimetry and Gorlin formula 0.80. Concordance analysis (Bland and Altman's method) gave mean (Mn) values for the differences in the areas determined by the Gorlin formula and the continuity equation of 0.01 +/- 0.15 cm2 (Mn - 2SD = -0.29, Mn + 2SD = 0.30). The estimated value by the Gorlin formula and planimetry was 0.02 +/- 0.10 (Mn - 2SD = -0.19, Mn + 2SD = 0.23). CONCLUSIONS: 1) Planimetry of the valve orifice area by transesophageal echocardiography is feasible and does not prolong the duration of the exam in the majority of patients. 2) The strong correlation and the results of concordance analysis, in the determination of valve orifice area, between traditional invasive methods and planimetry, support the use of this noninvasive method in clinical practice. PMID- 12090129 TI - Modulation of myocardial function by pyruvate. AB - OBJECTIVES: Pyruvate is an intermediate product of glycolytic metabolism that has a positive inotropic effect in animal models and in failing human hearts. The main objective of the present work was to clarify the mechanisms underlying this inotropic effect. METHODS: Isotonic and isometric twitches were recorded before and after the addition of pyruvate (3, 10 and 15 mM) to rabbit papillary muscles (n = 10) and human atrial trabeculae (n = 6) degrading glucose 9 mM and acetate 5 mM as metabolic substrates. In another protocol, undertaken in rabbit papillary muscles (n = 8), pyruvate was added in the presence of an inhibitor of mitochondrial pyruvate uptake (alpha-cyano-4-hydroxycinnamate [HCm]; 0.5 mM), and using octanoate 5 mM as metabolic substrate. Calculated parameters: active tension (AT); maximum velocity of tension rise (dT/dtmax); maximum velocity of tension decline (dT/dtmin); peak shortening (PS); maximum velocity of shortening (dL/dtmax); maximum velocity of lengthening (dL/dtmin) and time to half relaxation (tHR). Values are presented as means +/- SEM. RESULTS: In rabbit papillary muscles, pyruvate caused an initial transient negative inotropic effect (maximum at 3 min), followed by a sustained increase in myocardial contractility that stabilized 15 min later. The maximum negative inotropic effect was observed with 3 mM of pyruvate, decreasing AT by 13 +/- 4%, dT/dtmax by 14 +/- 5%, dT/dtmin by 9 +/- 3%, PS by 13 +/- 4% and dL/dtmax by 13 +/- 5%. Maximum positive inotropic effect was observed with 10 mM, which increased AT by 45.0 +/- 9.5%, dT/dtmax by 20.5 +/- 7.4%, PS by 33.4 +/- 9.6%, dL/dtmin by 35.5 +/- 12.1, and tHR by 27.8 +/- 3.2%, without significantly altering dL/dtmax or dT/dtmin. In the presence of HCm, the positive inotropic effect was not only observed but even enhanced. In human atrial trabeculae the addition of pyruvate also induced a similar increase in contractility, but the transient negative inotropic effect was absent. CONCLUSIONS: The addition of pyruvate caused a dose dependent positive inotropic effect observed in rabbit papillary muscles as well as in human atrial trabeculae. The effect of pyruvate in rabbit papillary muscles does not depend on its mitochondrial uptake and metabolism. This may be particularly relevant during myocardial ischemia when pyruvate concentration is increased and mitochondrial function is impaired. These characteristics give pyruvate a suitable profile for the metabolic protection of the heart. PMID- 12090130 TI - Subaortic stenosis diagnosed in the course of a twins pregnancy: a controversial management. AB - Fixed subaortic stenosis, commonly associated with other congenital cardiac defects, is the cause of 10 per cent of cases of congenital obstruction of the left ventricular outflow. Corrective surgery is frequently a successful treatment, recommendations being based on the transaortic gradient in Europe while in the USA the most prevalent opinion is surgical repair independently of the gradient. We present a case of adult clinical onset of a fixed subaortic stenosis during pregnancy, in which hemodynamic changes are significant, that was medically treated and followed in the outpatient clinic of our hospital, and review the state of the art of the management and surgical indications of this condition. PMID- 12090131 TI - Basics of methodologic analysis of therapeutic trials. Part I: Validity of the results. AB - One of the most compelling questions a cardiologist (and, for that matter, any clinician) has to face involves choosing the optimal management strategy for their patients. One of the best sources of information about therapy is the randomized controlled trial, which, despite several significant shortcomings, is considered to be the gold standard in studies about therapeutics and prevention. The methodological quality of therapeutic trials is highly variable and--as already discussed in previous articles--in order to be useful they have to be critically appraised in terms of the validity of their results, their importance, and their applicability to the individual patient. In this article, we will introduce guidelines for the critical appraisal of evidence in therapy, discussing the issue of validity. In future articles we will present a critical appraisal of the importance and applicability of randomized clinical trial results. PMID- 12090132 TI - Multiple intracavitary masses in an AIDS patient. Echocardiographic evolution. PMID- 12090135 TI - [Speech of the president of the Portuguese Society of Cardiology in the opening session -- April 14, 2002]. PMID- 12090136 TI - [23rd Congress of the Portuguese Society of Cardiology]. PMID- 12090137 TI - [Theoretical and physical aspects of excimer laser trabeculotomy (ELT) ab interno with the AIDA laser with a wave length of 308 mm]. AB - Ablation of tissue structures containing and surrounded by water differs from tissue ablation at a surface, not only theoretically, but also in terms of outcome. In contrast to the situation often observed with surface ablation, it is shown that the trabecular meshwork in the anterior chamber of the eye can be ablated cleanly and accurately with the 308 nm Excimer laser without causing collateral thermal damage. The reason for this is that in the trabecular meshwork, the ratio of radiation-absorbing tissue to water--which absorbs very little energy--is very small. A marked cooling effect thus results, which permits the development of only a very small amount of collateral thermal damage at the boundaries of the ablation zone. PMID- 12090138 TI - Detection of differentially expressed genes in particle disease using array filter analysis. AB - The precise cellular mechanism of osteolysis in particle disease is still unknown. The aim of the study was to screen for new gene products in macrophages during particle contact. METHOD: In an established macrophage model THP1-cells (human monocytic cells) were differentiated under the influence of vitamin D3 and GM-CSF into macrophage-like cells (MLC). MLCs were incubated each with different concentrations of polyethylene particles, Lipopolysaccharids (LPS) and controls. Isolated RNA was transcribed into complementary radioactive 32P labeled cDNA. This probe was hybridised on an human cDNA expression array and analysed by autoradiography. To obtain a more reliable method quantifying mRNA, the reverse transcriptase polymerase chain reaction (RT-PCR) was used. RESULTS: The arrays showed an upregulation of the following genes by particles: TNF-Rezeptor 2, IL-1 Receptor Antagonist, Bone Morphogenic Protein 4 and HM 145. This was proven three times using RT-PCR and statistically significant in comparison to the controls. LPS induced the same upregulation except for HM145 whereas particles caused downregulation of this mRNA expression. CONCLUSION: Our results prove that the model of differentiated THP-1 cells treated with PE particles is a suitable system to analyse differential gene expression patterns, since the induction of the major positive control genes TNF alpha and IL1 beta were detected by this approach. BMP 4 is known as signal protein which mediates ectopic bone formation and can also be interpreted as a contra regulatory gene. HM 145 belongs to the leukocyte chemotactic peptide receptor family. HM 145 seems to be one of the first genes that is enhanced along the septical pathway but less expressed by contact with particles. Analysis of HM 145 expression might help to diagnose septic versus aseptic loosening of prosthesis. PMID- 12090139 TI - High frequency intra-QRS signals in idiopathic dilated cardiomyopathy. AB - We extracted and quantified high frequency intra-QRS signals in idiopathic dilated cardiomyopathy (IDC). In IDC the analysis of late potentials in the terminal QRS complex often fails in predicting clinical events because of intraventricular conduction abnormalities and the absence of a circumscribed arrhythmogenic substrate. Therefore, new approaches are required to assess the electrical state of the myocardium. We investigated 21 patients suffering from IDC with (n = 14) and without (n = 7) bundle branch block. High resolution 31 lead magnetocardiograms were filtered with a 67 point 4th order Savitzky-Golay filter. The difference of the measured and filtered signals was calculated (67 200 Hz). The spatio-temporal properties and the areas under the curves of the resulting high frequency intra-QRS signals (IQCs) were studied. We detected IQCs in all patients. The patients had individual patterns regarding the temporal and spatial properties of the IQCs during depolarisation. The IQCs predominantly appeared in the initial portion of the QRS. The ratios of the areas under the curves of the IQCs and the measured signals were linearly correlated to the left ventricular enddiastolic diameter (r = 0.71, significance 0.0012). In IDC the ventricular depolarization is accompanied by individual spatial and temporal patterns of high frequency intra-QRS signals. They can be studied non-invasively from body surface mapping data with the algorithm used in this study. This provides access to the assessment of the electrical status in patients with IDC. PMID- 12090140 TI - [Effect of a double intervertebral cage on the mechanical behavior of the lumbar spine]. AB - A three-dimensional nonlinear finite element model of the lumbar spine was developed. Paired threaded cages and a monosegmental internal spinal fixation device were integrated into the computer model. The model was loaded with such forces as apply during standing, as well as with pure moments in the three main anatomical planes, plus an additional preload. The latter was generated by shortening the distance between the pedicle screws on the longitudinal rod of the fixator. With the exception of torsional loading, an implant appreciably reduces the mobility in the segment concerned. At the loads studied, cages had only a minor impact on the movements and stresses in the adjacent regions, but a strong influence on the stresses in the endplates in contact with them. A preload increases these stresses dramatically. Contact conditions between vertebral body and cages also have a marked effect on the stress distribution in the corresponding vertebral endplate, especially in the case of extension loading. Owing to the preload, maximum stresses were higher for the rigid bond than when contact elements were used. PMID- 12090141 TI - [Measuring ligament elasticity of the knee joint--elasticity measuring strip and its alternatives]. AB - Those techniques for measuring ligament tension at the knee joint that are most commonly cited and easiest to carry out are discussed. These include four techniques based on the use of strain gauges. Apart from the Omega transducer and the buckle transducer, there is also the tendon force transducer, and the application of strain gauges to the bony ligament insertion sites. Other indirect measuring methods considered are the mercury strain transducer and the Hall effect transducer. The parameter measured with all of these methods is fluctuating current or voltage, which is then correlated with ligament tension. Three direct measurements are also discussed: the separation distances of marked fibres of the ligaments, replacement of fibres by threads, and a load cell/bone plug construction. The measured value is equated with the effective change in ligament length. PMID- 12090142 TI - [Correlation of electromyography and clinical results after bicondylar knee joint prosthesis implantation]. AB - AIM OF THE STUDY: To investigate clinical findings in patients receiving bicondylar sledge prostheses, and their correlation with electromyographic measurements in comparison with healthy volunteers. MATERIAL AND METHODS: An average of 31.9 months after implantation of a bicondylar sledge prosthesis, 15 patients were clinically examined and compared with 11 control subjects. Electromyographic measurements of the rectus femoris, vastus medialis and lateralis, semitendinosus, biceps femoris, tibialis anterior, and gastrocnemius muscles were obtained. RESULTS: All clinical scores were significantly poorer in the patient group. The activity of the vastus lateralis was significantly reduced. The activity in the lower leg of the operated side was significantly higher than in the control group. Significant differences were found in the tibialis anterior and gastrocnemius muscles between controls and patients. Analysis of the correlation between the clinical scores and the electromyographic measurements showed no significance. CONCLUSION: The clinical results correlated only slightly with the electromyographic activity of the muscles investigated. The commonly presumed quadriceps insufficiency was not confirmed. PMID- 12090144 TI - [The three pillars of clinical pharmacology]. PMID- 12090143 TI - [A cost-effectiveness analysis of changes in therapeutic strategies in the treatment of HIV since 1996]. AB - The objective was to assess the cost-effectiveness ratio of HAART in the treatment of HIV infection. Two random samples were extracted from the database of the Rothschild Public Hospital, and patients were matched for age, sex and T4 cell counts: a first sample selected in 1996/97 of HAART treated patients (CAS group) and a second sample selected in 1994/95 of non-HAART treated patients (CONTROL group). Immune recovery and use of resources data were extracted and analyzed over two years for 196 included patients. Mean T4 cell count after two years was higher among CAS patients (344/mm3 vs. 234/mm3; p < 0.0001). CAS patients recorded a supplementary cost of antiretroviral treatments (+171%; p < 0.0001) balanced by savings in other drugs expenses (-62%; p = 0.0560) and in hospitalizations (-25%; NS). Overall, CAS patients presented a 15% (NS) lower medical cost than CONTROL patients. PMID- 12090145 TI - [Desirudin (Revasc) to prevent thromboembolic complications after hip or knee replacement surgery]. AB - Since March 1999, desirudin (REVASC), a recombinant hirudin, has been used in Nancy to treat patients who undergo total hip or knee replacement with a high risk of thromboembolic complications. We carried out a retrospective study using clinical data on the first 15 consecutive patients treated with desirudin to find out prescription motivations, type of shift (indirect anticoagulants or low molecular-weight-heparin) and evolution. They all had a high risk of deep vein thrombosis (thrombophily, obesity, history of thromboembolic events). Some of this patients would have been excluded of the studies which permitted desirudin to be approved. In this study, we found no thromboembolic complications. The only striking facts are one bleeding complication (after difficult surgery) and one pulmonary embolism (2 months later). PMID- 12090146 TI - [Evaluation of the impact of individual antibiotic order forms on consumption of antibiotics]. AB - The aim of the study was to assess the impact of an individual patient order form which concerned the 21 most costly antibiotics in a university hospital. Antibiotics expenditures were monitored from 1995 to 1999 and were expressed in 1999 French Francs per 100 patient days (p.d.). The time series were analyzed by auto-regressive models. The trend of antibiotics expenditures which were concerned by the individual patient order form was a yearly increase of 50 FF/100 p.d. (p < 0.01). The individual patient order form had no significant impact on global antibiotics expenditures but there were some differences across departments: antibiotics costs decreased 1.293 FF/100 p.d. (p = 0.02) in intensive care departments. Monitoring antibiotics consumption should be continued in order to increase power of analysis and to assess the impact of the implementation of guidelines. PMID- 12090147 TI - [An adverse effect of glycine irrigation solution: absorption syndrome]. AB - Endoscopic operations in the genitourinary tract require the use of an irrigating fluid containing glycine 1.5% and expose patients to adverse events in relation with the absorption of that irrigating fluid, summarized as "transurethral resection reactions" (TURP syndrome). Intra and extra vascular absorption are now well described. The uptake of 1000 ml of fluid corresponds to an acute decrease in the serum sodium concentration of 5-8 mmol.l-1 and give poor clinical signs. This is the volume above which the risk of absorption-related symptoms is statistically increased. Dilutional hyponatraemia and toxicity of glycine and/or its metabolites explain the clinical symptoms. Hyponatraemia and the osmotic gap assess the diagnosis of TUR syndrome. There is a threat to life and a knowledge of an adequate management is essential. Ethanol monitoring is an non invasive method to prevent and detect early irrigating fluid absorption. PMID- 12090148 TI - [Results of a French nationwide survey of cutaneous side effects of ketoprofen gel reported between September 1996 and August 2000]. AB - A French nation-wide pharmacovigilance survey of ketoprofen topic collated, from September 1996 to August 2000, 770 cutaneous side-effects. The frequency varies from 0.013@1000 to 0.028@1000 according to the commercial gel. Analysis concerns spontaneous notifications of French nation-wide pharmacovigilance. Sex ratio is well distributed, population is young. Treatment lasted about 12 days, the side effects appearing after about 13 days, 25 per cent of cases are delayed to discontinuance of treatment, mainly after exposure to sunlight. Co-administered drugs are in most cases systemic or topical NSAIDs and/or fibrates and then increase the seriousness of the iatrogenic pathology. 75 per cent of cutaneous side-effects appear in summer, 50 per cent have been reported as "photosensitivity". Reactions are severe in 30 per cent of the cases. More than 80 per cent of cases present an extension beyond the site of application. The course is usually favourable and neither topical nor systemic corticosteroid treatment influence the duration of evolution. Photopatchtests testing in 23 per cent of cases show evidence of photoallergy to ketoprofen and crossed photoallergy with tiaprofenic acid, fenofibrate, oxybenzone and benzophenone. These results confirm that photoallergy is due to the common benzoylketone structure but not to their arylpropionic function. Some cases of persistent or recurrent photosensitivity must be more explored. The results lead to request a modification of marketing authorizations. PMID- 12090149 TI - [The origins of clinical pharmacology in France]. AB - This text is a history of the early development of clinical pharmacology in France in the 60s and 70s. It relates the obstacles that needed to be surmounted to impose clinical trials, pharmacovigilance, therapeutic drug monitoring. It follows the evolution of regulations, and the constitution of a new hospital and academic discipline. This study shows the obstacles raised by an established medical tradition against a change in paradigm towards a scientific approach to the evaluation and clinical use of drugs. It relates a crucial moment of the history of French pharmacology, when French medicine entered the modern era. PMID- 12090150 TI - [Tolerance of riluzole in a phase IIIb clinical trial]. AB - Within the framework of an early drug access programme launched in 1995, a multicentre open study was initiated in France in order to assess, inter alia, the safety of riluzole (50 mg twice a day) in a total of 2069 patients from 28 centres. This programme, a phase IIIb study with direct individual benefit, had two main objectives: to enable patients to receive riluzole therapy pending regulatory approval and commercial availability and to provide further data on the safety of riluzole in a broader ALS population. The most frequent adverse events related to riluzole treatment were: asthenia, nausea and elevation of serum transaminase levels. These observations, similar to data derived from previous pivotal clinical trials, confirm that riluzole has a satisfactory tolerability profile. PMID- 12090151 TI - [Oral antibiotic therapy in current practice: acute otitis media]. PMID- 12090152 TI - [Oral antibiotic therapy in current practice: lower respiratory tract infections in children]. PMID- 12090153 TI - [Oral antibiotic therapy in current practice: acute sinusitis in children]. PMID- 12090154 TI - [Oral antibiotic therapy in current practice: acute sinusitis in adults]. PMID- 12090155 TI - [Oral antibiotic therapy in current practice: exacerbation of chronic bronchitis]. PMID- 12090156 TI - The relation between xerostomia and hyposalivation in subjects with rheumatoid arthritis or fibromyalgia. AB - Aim of this study was to evaluate the relation between xerostomia and hyposalivation in 100 subjects with either rheumatoid arthritis or fibromyalgia, and further, to evaluate the predictive value of xerostomia on hyposalivation. Unstimulated and chewing stimulated whole saliva was collected in the morning with the subjects in a strict fasting condition and then about 2 hours later, after intake of a standardised breakfast. All participants filled in a questionnaire, mainly dealing with xerostomia. Forty subjects demonstrated a pathological fasting unstimulated whole saliva secretion rate, the corresponding number for fasting stimulated secretion being 39. For unstimulated, but not for stimulated saliva, the fasting secretion rate was significantly lower than the non-fasting. Xerostomia was reported by 74 subjects, this group having significantly lower both unstimulated and stimulated secretion rates than the non xerostomic group. On the individual level, the predictive value of xerostomia on hyposalivation showed high sensitivity but unsatisfactory specificity. In conclusion, this study underlines the importance of applying strictly standardised procedures when collecting saliva, and that fasting unstimulated whole saliva is the diagnostic salivary secretion of choice. Finally, xerostomia was found to predict hyposalivation on a group, but not on an individual level. PMID- 12090157 TI - Marginal bone loss and tooth loss in a sample from the County of Stockholm--a longitudinal study over 20 years. AB - The purpose of the study was to assess the degree of marginal alveolar bone loss and tooth loss in a longitudinal study over a period of 20 years. In 1970 a medical and odontological study on a stratified sample of the population of the County of Stockholm aged 18-65 years started. Twenty years later a new odontological investigation was performed on the 513 subjects who wanted to take part once more and who had at least five remaining teeth. The mean distance between the marginal bone level and the cemento-enamel junction measured on the radiographs in 1970 (MBL) was 13.8% of the root length. The mean bone loss during the 20-year period (MBD) was about 10%, corresponding to a mean annual marginal bone loss of about 0.1 mm. The MBD-values were significantly positively correlated to the MBL-values. The mean number of remaining teeth was 24.7 in 1970 and 21.8 in 1990. A significant positive correlation existed between the number of teeth lost between 1970 and 1990 and the mean MBD-value. In stepwise multiple regression analyses, the MBL-value was the only independent variable positively correlated to the MBD-value, while the number of teeth lost between 1970 and 1990 was significantly correlated to age and the MBL-value. PMID- 12090158 TI - Objective and subjective need for orthodontic treatment in Stockholm County. AB - The objective and subjective need for orthodontic treatment according to a modified version of the treatment priority index of the Swedish National Board of Health and Welfare (index scores 0 to 4), was studied in 1281 children and teenagers, aged between 8 and 16 years and living in the Stockholm area. Furthermore, differences between different parts of the Stockholm County with respect to need for orthodontic treatment, and the subjective demand for treatment were also recorded. Five areas with varying population structures were studied. The results showed that the prevailing objective and subjective need for treatment varied in the areas studied. The prevalence of objective need for treatment varied between 23.8% and 28.9%. The subjective need for treatment ranged from 21.6% to 30.3%. The highest prevalence of treatment need was found in areas with a large immigrant population. In all the investigated areas was the subjective demand for treatment 1-1.5% less than the objective need for treatment. On the basis of the present data, it can be concluded that neither the objective, nor the subjective need for orthodontic treatment exhibits any tendency to decline. The results do not, therefore, support the policy that the public financial resources for orthodontic treatment could be reduced due to a lowered need for treatment. Severe malocclusions may result in impaired craniomandibular function and have an unfavourable influence on facial and dental attractiveness, which may in turn have psychosocial implications. There is an obvious risk that the frequency of such problems may increase in the population, if children with great or very great need for treatment are denied adequate orthodontic care due to lack of sufficient resources. PMID- 12090159 TI - Determinants of costs of orthodontic treatment in the Finnish public health service. AB - Factors affecting the costs of children's free orthodontic treatment were studied from patient records of a random sample of 193 successfully treated orthodontic patients, aged 7-14 years at the start of the treatment. They were treated by orthodontic specialists in the health centers of the cities of Joensuu, Oulu, and Vaasa. The statistical analysis was done using advanced regression analysis, two tailed t-test and chi-square test. The results indicated that the number of appliances used, the patients' ages at the start of treatment, the number of missed appointments, differences between health centers, and the change in PAR (Peer Assessment Rating) Index scores accounted for 78.7% of the variations in treatment costs. The number of appliances used was the most important factor in the regression (R2 = 0.657). The costs of treatment were lowest in one-stage treatments started in the permanent dentition, and were highest in two-stage treatments started in the mixed dentition. Significant differences were found in treatment patterns and costs between the three health centers, reflecting low concern for cost-efficiency aspects. PMID- 12090160 TI - Prevalence of different temporomandibular joint sounds, with emphasis on disc displacement, in patients with temporomandibular disorders and controls. AB - Temporomandibular joint (TMJ) sounds are very common among patients with temporomandibular disorders (TMD), but also in non-patient populations. A variety of different causes to TMJ-sounds have been suggested e.g. arthrotic changes in the TMJs, anatomical variations, muscular incoordination and disc displacement. In the present investigation, the prevalence and type of different joint sounds were registered in 125 consecutive patients with suspected TMD and in 125 matched controls. Some kind of joint sound was recorded in 56% of the TMD patients and in 36% of the controls. The awareness of joint sounds was higher among TMD patients as compared to controls (88% and 60% respectively). The most common sound recorded in both groups was reciprocal clickings indicative of a disc displacement, while not one single case fulfilling the criteria for clicking due to a muscular incoordination was found. In the TMD group women with disc displacement reported sleeping on the stomach significantly more often than women without disc displacement did. An increased general joint laxity was found in 39% of the TMD patients with disc displacement, while this was found in only 9% of the patients with disc displacement in the control group. To conclude, disc displacement is probably the most common cause to TMJ sounds, while the existence of TMJ sounds due to a muscular incoordination can be questioned. Furthermore, sleeping on the stomach might be associated with disc displacement, while general joint laxity is probably not a causative factor, but a seeking care factor in patients with disc displacement. PMID- 12090161 TI - [Strong lobbying for direct -to-consumer advertising of prescription drugs. Beneficial for the patient or for the increase of sales?]. PMID- 12090162 TI - [Bibliometric data result in wrong interpretation of the scientific creativeness]. PMID- 12090163 TI - [Several cases of tick-borne encephalitis in Skane. Should high-risk groups be vaccinated?]. AB - Three cases of tick-borne encephalitis from the non-endemic southern Swedish region of Skane is reported. The clinical course was moderate to severe including neurological sequelae. The question is raised whether risk groups also in southern Sweden should be vaccinated. PMID- 12090164 TI - [A new putative principle for diagnosis of Creutzfeldt-Jakob disease]. AB - Creutzfeldt-Jakob's disease and bovine spongiform encephalitis (BSE) are both prion diseases, i.e., diseases caused by an abnormally folded isoform of cellular prion protein. A variant of Creutzfeldt-Jakob's disease can probably be transmitted from cattle with BSE to humans. To prevent spread of BSE, whole stocks of cattle are destroyed when symptoms of the disease appear. However, this is too late to prevent transmission during the about 5 years long incubation time. A method for presymptomatic diagnosis of BSE is clearly desirable. Miele and colleagues at Roslin Institute in Edinburgh present a potential molecular marker for prion diseases in the March issue of Nature Medicine. It is a dramatically decreased expression of a transcript called erythroid differentiation-related factor (EDRF). This change is detectable early in the course of the disease also in tissues outside the central nervous system, for instance blood. PMID- 12090165 TI - [Heart failure in amyloidosis--a rare but treatable disease]. AB - Cardiac amyloidosis is rare, but should be considered in the differential diagnosis of any patient with congestive heart failure who does not have evidence of ischemic heart disease. The diagnosis is more likely in patients with increased wall thickness and normal chamber size on echocardiogram, low voltage on the electrocardiogram, and evidence suggesting a multisystem disease. Amyloid proteins are varied; the most common are AA, AL and transthyretin. The clinical features differ and are largely determined by the underlying disease responsible for the amyloid infiltration and of the organs involved. The specific type of protein must be determined as the prognosis and treatment vary among the different types of amyloidoses. Systemic amyloidoses are life-threatening diseases but making an early diagnosis in connection with today's more specific treatment may modify the course of the disease. PMID- 12090166 TI - [Six case-control studies from the Osterbro-study (The Copenhagen City Heart Study). Angiotensinogen mutations and risk of myocardial and cerebral ischemia]. PMID- 12090167 TI - [Growth of insulin-producing beta cells. Perhaps a new way to treat type 1 diabetes]. AB - Insulin dependent diabetes mellitus (type 1-diabetes) results from a selective destruction of the insulin producing beta cells and a limited capacity of the remaining cells to regenerate in a compensatory manner. Increased knowledge of the factors involved in the regulation of beta-cell growth may lead to new ways of forming beta cells that in combination with selective immunosuppression could be used for treatment of type 1 diabetes. Physiological and pathophysiological conditions that induce beta-cell growth, different factors that have been implicated to regulate these processes and future perspectives concerning treatment of type 1 diabetes by expanding the beta-cell mass is discussed in this review article. PMID- 12090168 TI - [Mastocytosis--two diseases with different physiopathology]. AB - Mastocytosis means proliferation and accumulation of mast cells in skin and internal organs. Bone marrow cytogenetic abnormalities are similar to those found in myeloproliferative diseases. Recent findings indicate different pathogenetic forms of mastocytosis. Thus, adult patients with associated hematological diseases express mutation of the gene for the thyrosine kinase receptor, while childhood cases lack this mutation. In adults the skin and bone marrow are the most commonly affected organs but involvement of the skeleton and gastrointestinal tract also occur. Hematological malignancy is a rare but well recognized complication. Childhood onset mastocytosis in the skin regresses spontaneously, whereas virtually all of the adult-onset cases persist. PMID- 12090169 TI - [A case report. Calf muscle infarction--a diagnostic problem!]. PMID- 12090171 TI - [The bilingual brain]. PMID- 12090170 TI - [Irinotekan in spread small cell lung cancer. More studies are required to change the practice]. PMID- 12090172 TI - [Discrimination of elderly patients in clinical research and health care]. PMID- 12090173 TI - [The society of hospitalists: mutual task for physicians and politicians to explain community-based health care]. PMID- 12090174 TI - [Obvious advantages with insurance-financed health care]. PMID- 12090175 TI - [Economic control in psychiatry--failures and possibilities]. PMID- 12090176 TI - [Remarkable contribution on estrogen, part 2]. PMID- 12090177 TI - [WHO proposals on child health in the 21st century. Treat and prevent the big "child killers"]. PMID- 12090178 TI - [Ethical aspects of the meeting with the obese patient: Why don't we see the obesity?]. PMID- 12090179 TI - [Laws concerning the psychiatric involuntary commitment should be available in several languages, not only in Swedish and English]. PMID- 12090180 TI - [Doping--time for reflection and reconsideration!?]. PMID- 12090181 TI - [Development of the ESCAP-CDV as measuring tool for the assessment of quality of life in prostatic cancer]. AB - The current following of oncological patients is based in a biopsychosocial model where in addition to evaluate the response to treatment, the interval free of illness and the global survival it includes the functional capacity and the psychosocial charge the illness causes in patients. Nowadays, different treatments have improved the survival, therefore the number of years of inability increase. It creates a new variable, the quality of life related to health (QLRH) that valves the individual respond to the physical, mental and social effects that health or illness produces on patients daily life. To establish the QLRH as an instrument of measure is needed. In Spain there isn't any validated instrument for the evaluation of it in patients with prostate cancer. We have developed the ESCAP-CDV (Quality of Life study on prostate cancer) an instrument elaborated by an interdisciplinary group made of urologist, psichologists and a doctor specialized in family medicine expert in psychometric validation of scales. We evaluate multiple dimensions of quality of life in a questionnaires autoadministed. PMID- 12090182 TI - [Posterior sacral root neuromodulation in the treatment of chronic urinary dysfunction]. AB - OBJECTIVES: To describe the effectiveness of sacral root neuromodulation in ameliorating symptoms of refractory voiding disfunction in our center. MATERIAL AND METHODS: During the period from december 1998 throught december 2001, 31 Percutaneous Nerve Evaluation (PNE) was performed to 20 patients with refractory voiding dysfunction; a sacral nerve stimulation device was implanted in 10 patients (8 female, 2 male). The median age was 49 years. Refractory voiding dysfunction included: mixed disorders (30%), idiopathic nonobstructive chronic urinary retention (20%), urgency/frequency (20%), faecal and urinary incontinence with absence of sphincter defect (20%) and frequency (10%). RESULTS: The 2 patients with frequency/urgency decreased their symptoms more than 50%, bladders were emptied without post-void residual urine in 2 patients with urinary retention, faecal and urinary incontinence in 2 patients and mixed disorders in other 3, reduced their symptoms more than 90% without residual urine. The frequency improved more than 50% in 1 patient. CONCLUSIONS: Sacral Root Neuromodulation is a successful treatment in some cases of idiopathic chronic micturition dysfunction which don't respond to pharmacotheraphy or bladder retraining. The effects of neuromodulation are long-lasting and associated morbidity is low. PMID- 12090183 TI - [Our experience with the implant of single component integrated prostheses (Hydroflex-Dynaflex)]. AB - INTRODUCTION AND OBJECTIVES: To present our experience with Hydroflex and Dynaflex self-contained inflatable penile prosthesis. MATERIAL AND METHODS: Between october 1988 and december 2000 a total of 63 men underwent implantation of 8 Hydroflex and 55 Dynaflex penile prosthesis. The follow-up period ranged from 12 and 127 months (mean 54.5). RESULTS: A total of 124 cylinders were implanted (in 2 cases only 1 cylinder was implanted). During implantation 3 corpora cavernosun ruptures were present, but implantation was possible in 2. Specific complication rates after implantation were as follow: early infection, 3 patients (4.7%); late infection, 1 patient (1.6%); pain that needed readmission 2 cases (3.2%); mechanical device failure, 7 cylinders in 6 men (mean time 83.5 months). 7 men (11.1%) needed prolonged teaching time to operate the prosthesis. Only 6 men (9.5%) wer dissatisfied with the prosthesis. It was possible to change only 1 cylinder when needed. CONCLUSIONS: Self-contained inflatable prosthesis brings good results on long time use. The mechanical malfunction rate is low and it is possible to change only one cylinder if needed. An important number of patients need intensive and prolonged teaching time to obtain a successful result. PMID- 12090184 TI - [Half-stay and morbidity in a cohort of patients treated with urologic surgery iin the treatment with acenocumarol (Sintrom)]. AB - OBJECTIVE: Our objective was to evaluate the effect about half stay at hospital, in patients treated with Sintrom, which required different urological surgery. PATIENTS AND METHOD: We evaluated 55 patients in treatment with Sintrom which required different urological surgery. We analysed a cohort control group with similar characteristic without anticoagulation therapy. The patients was at hospital 3 days before surgery to realize the change Sintrom/Heparina. We used the haematology service's protocol at the Hospital del Mar. We analysed in each patient the half stay at hospital and the morbidity referred to bleeding complications. The decrease of haemoglobin higher than 2 g/dl was considered significant. RESULTS: About half stay, all patients required more days of hospitalization. The additional morbidity was obvious in the transurethral resection of the prostate and in great not endoscopical surgery. In the rest of surgery the complications referred to bleeding have repercussion about stay at hospital. CONCLUSIONS: We found a significant increased half stay at hospital and the morbidity referred to bleeding complications and its consequence (transfusion, second surgery,...). PMID- 12090185 TI - [Detection of prostatic cancer in the PSA range between 3 and 3.9 ng/ml]. AB - OBJECTIVE: In our study, we analyze the benefit of lowering the PSA cutoff point for which a prostate biopsy is indicated from 4 to 3 ng/ml. MATERIALS AND METHODS: We have considered 4.278 individuals coming from a prostate cancer screening program. We studied 1.217 interventions in which PSA was determined, indicating the prostate biopsy with PSA > or = 3 ng/ml. Digital rectal examination was never the indication for the biopsy. All biopsies were sextant and assisted by transrectal ultrasound. We compared the performance of the biopsy using 4 and 3 ng/ml as cut points. RESULTS: Of the 1.217 interventions performed, 947 had PSA values lower than 3 ng/ml, 80 between 3 and 3.9 ng/ml and 190 over 4 ng/ml. A total of 189 patients (70% of these two last groups) underwent a prostate biopsy. With 4 ng/ml as the cut point, 134 biopsies were indicated, detecting 28 cancers (positive predictive value 20.9%). However 189 biopsies were indicated and 34 cancers detected by lowering the cut point to 3 ng/ml (positive predictive value 17.9%). The reduction in the biopsy performance was not statistically significant (OR = 0.89). None of the 6 additional cancers detected was palpable or ecographically visible (T1c), all of them had a Gleason score under 7 and half of them could be considered clinically relevant. CONCLUSIONS: Lowering PSA cutoff point from 4 to 3 ng/ml improved the detection rate in 21.4% not jeopardizing the biopsy performance. Therefore we think that the group of patients with PSA between 3 and 3.9 ng/ml as candidates for prostate biopsy, should be included in screening programs. PMID- 12090186 TI - [Effects of peripheral functional electrostimulation in obstructive bladder instability]. AB - INTRODUCTION: The aim of this study was to assess the effects of peripheral functional electrical stimulation (FES) in patients with detrusor instability (DI) secondary to obstructive Benign Prostatic Hyperplasia (BPH). SUBJECTS, MATERIAL & METHODS: Nine patients with urodynamically proven DI secondary to BPH were investigated. Sensory and pudendo-anal reflex thresholds were determined in each patient to individually optimise FES. A standard fill cystometry was then performed and individually optimised FES was given when an unstable detrusor contraction occurred. Non stimulated contractions were used as controls. The effects of FES were analysed by measuring the mean area under the unstable pressure curves with and without FES and were expressed as percentage suppression. RESULTS: The percentage suppression in the mean area under the curves after stimulation was < 10%. There was no statistically significant difference between controls and FES using a two tailed, paired students t-Test (p > 0.1) at 95% confidence level (p > or = 0.05). CONCLUSION: FES did not significantly suppress unstable bladder contractions in patients with DI secondary to BPH. This finding suggests that obstructive DI does not depend on sacral reflex mechanisms and supports the view that it has a peripheral pathophysiological origin (myogenic and/or nerve endings). PMID- 12090187 TI - [Coordinated expression of cytokeratin 7 and 20 in transitional carcinoma of the bladder: diagnostic usefulness]. AB - BACKGROUND: Cytokeratins are intermediate filament proteins which are part of the framework of eucariotic cells. A characteristic pattern of expression of cytokeratins has been described for each type of epithelium. This pattern is maintained during the process of malignancy and metastasis. Immunohistochemical analysis of cytokeratins 7 and 20 (Ck 7 and Ck 20) has been considered particularly useful for identifying the origin of carcinomas. METHODS: A total of 122 cases of transitional cell carcinoma of urinary bladder were studied immunohistochemically with monoclonal antibodies against Ck 7 and Ck 20. The peroxidase labelled-streptavidin biotin technique was performed. Cases were considered positive when at least 1% of tumoral cells were stained. RESULTS: All cases investigated expressed Ck 7 whereas Ck 20 was positive in 75% of them. Reactivity for Ck 20 was always focal. No tumor was negative for Ck 7. CONCLUSION: Ck 7+/Ck 20+ immunophenotype is highly characteristic of transitional cell carcinoma of urinary bladder, whereas Ck 7+/Ck 20- is less frequent. Immunohistochemical determination of these cytokeratins is useful for the diagnosis when tumors are poorly differentiated and for identifying the primary site of metastatic carcinomas. PMID- 12090188 TI - [Laparoscopic diagnosis of Hodgkin's disease. Report of a case]. AB - Laparoscopic Surgery with diagnosis objectives in Urology, has many others indications related with different medical specialties, apart from the ones for the urologic area (ectopic test, linfadenectomy, etc...). In our case, the Services of Urology and Hematology, diagnosed a Hodgkin's disease case. The Service of Hematology realized treatment and evolution. Sometimes, the unspecific clinical presentations of the Hodgkin's disease, shows adenopatics masses, detected by Ecografy or TAC. Treatment and prognosis of the Hodgkin's disease, are related with the histological subtype and its spreading. PMID- 12090189 TI - [Oat cell carcinoma of the bladder]. AB - Small cell carcinoma (CCP) of the urinary bladder is an uncommon tumor, having usually an aggressive behavior. We report a new case of CCP seen at our centre and we communicate the applied therapeutic method. Although total cystectomy followed by chemo and radiotherapy seems to be the most effective treatment, we have carried out radical RTU of the bladder accompanied by chemo and radiotherapy as conservative therapy obtaining complete remission after 2 year of diagnosis, without local and distant evidence of the disease. PMID- 12090190 TI - [Bellini's duct carcinoma]. AB - Two new cases of uncommon Bellini's duct carcinoma are described. The existing literature is reviewed, discussing the clinical manifestations, diagnosis, treatment and embryological aspects, which differentiate from clear cell carcinoma. PMID- 12090191 TI - [Signet-ring cell neoplasm. Unusual histologic variety as primary bladder tumor]. AB - We report a case of primary signet-ring cell adenocarcinoma of the urinary bladder. It is a extremely rare tumor. Gross hematuria is the commonest clinical presentation as well as this case. Our patient presented with advanced tumour invasion into perivesical fat at dome. The treatment was partial cystectomy together with quimiotherapy. Medical literature are reviewed with special remark on histopathology and its bladder origin. PMID- 12090192 TI - [Leydig cell tumor. Two new cases]. AB - The Leydig's cell tumours are the most common of all non germ cell tumour of the testis, that clinical findings is different by age of presented. We report two new cases of Leydig cell tumour diagnosed in our centre having different presentation form, in old adults with feminizing features and occult tumour, and as a testicular nodule pain, respectively. The analytical, clinical, diagnostic aspects and therapeutic of the uncommon disease are discussed. PMID- 12090193 TI - [Bilateral obstructive uropathy secondary to inguinoscrotal bladder hernia]. AB - Bladder hernia is present in an important number of cases of inguinal hernias. Massive inguinoscrotal bladder hernias are rarer. The association of massive bladder hernia and bilateral ureterohydronephrosis is not often found. According to our knowledge, there have been only seven cases documented in the last ten years. Therefore, we will present a case of renal failure secondary to inguinoscrotal bladder hernia with bilateral obstructive uropathy. Once we have analyzed a clinical case, we will give the diagnosis and treatment for those hernias. PMID- 12090194 TI - [Unilateral agenesia of deferent duct]. PMID- 12090195 TI - The awakening!!! PMID- 12090196 TI - Complex emergencies: an introduction. PMID- 12090197 TI - Relevant in times of turmoil: WHO and public health in unstable situations. AB - For millions of people world-wide, surviving the pressure of extreme events is the predominant objective in daily existence. The distinction between natural and human-induced disasters is becoming more and more blurred. Some countries have known only armed conflict for the last 25 years, and their number is increasing. Recently, humanitarian sources reported 24 ongoing emergencies, each of them involving at least 300,000 people "requiring international assistance to avoid malnutrition or death". All together, including the countries still only at risk and those emerging from armed conflicts, 73 countries, i.e., almost 1.8 trillion people, were undergoing differing degrees of instability. Instability must be envisioned as a spectrum extending between "Utopia" and "Chaos". As emergencies bring forward extreme challenges to human life, medical and public health ethics make it imperative for the World Health Organisation (WHO) to be involved. As such, WHO must enhance its presence and effectiveness in its capacity as a universally accepted advocate for public health. Furthermore, as crises become more enmeshed with the legitimacy of the State, and armed conflicts become more directed against countries' social capital, they impinge more on WHO's work, and WHO must reconcile its unique responsibility in the health sector, the humanitarian imperative and the mandate to assist its primary constituents. Health can be viewed as a bridge to peace. The Organization specifically has recognised that disasters can and do affect the achievement of health and health system objectives. Within WHO, the Department of Emergency and Humanitarian Action (EHA) is the instrument for intervention in such situations. The scope of EHA is defined in terms of humanitarian action, emergency preparedness, national capacity building, and advocacy for humanitarian principles. The WHO's role is changing from ensuring a two-way flow of information on new scientific developments in public health in the ideal all-stable, all-equitable, well resourced state, to dealing with sheer survival when the state is shattered or is part of the problem. The WHO poses itself the explicit goals to reduce avoidable loss of life, burden of disease and disability in emergencies and post-crisis transitions, and to ensure that the Humanitarian Health Assistance is in-line with international standards and local priorities and does not compromise future health development. A planning tree is presented. The World Health Organization must improve its own performance. This requires three key pre-conditions: 1) presence; 2) surge capacity; and 3) institutional support, knowledge, and competencies. Thus, in order to be effective, WHO's presence and surge capacity in emergencies must integrate the institutional knowledge, the competencies, and the managerial set-up of the Organization. PMID- 12090198 TI - Complex emergencies: expected and unexpected consequences. AB - Complex emergencies emerged as a new type of disaster following the end of the Cold War, and have become increasingly common in recent years. Human activity including civil strife, war, and political repression often coexist with and contribute to natural phenomena such as famine. They frequently result in high mortality, population displacement, and the disruption of civil society and its infrastructure. This article reviews the context of recent complex emergencies, and their expected health consequences, such as diarrhea, measles, malnutrition and outbreaks of infectious disease, and the disruption of mechanisms of disease control and surveillance. However, the complex nature of these emergencies also may have unexpected consequences, such as hindering understanding of their causes or limiting the attention paid to them by the public. This paper discusses the context and consequences of complex emergencies from the health standpoint, and explores some of their unexpected effects. PMID- 12090199 TI - Military assistance in complex emergencies: what have we learned since the Kurdish relief effort? AB - After the success of relief efforts to the displaced Kurdish population in northern Iraq following the Gulf War, many in the US military and the international relief community saw military forces as critical partners in the response to future complex emergencies (CEs). However, successes in subsequent military involvement in Somalia, Rwanda, the former Yugoslavia, and other CEs proved more elusive and raised many difficult issues. A review of these operations reinforces some basic lessons that must be heeded if the use of military forces in humanitarian relief is to be successful. Each CE is unique, thus, each military mission must be clearly defined and articulated. Armed forces struggle to provide both security and humanitarian relief, particularly when aggressive peace enforcement is required. Significant political and public support is necessary for military involvement and success. Military forces cannot execute humanitarian assistance missions on an ad hoc basis, but must continue to develop doctrine, policy and procedures in this area and adequately train, supply, and equip the units that will be involved in humanitarian relief. Militaries not only must cooperate and coordinate extensively with each other, but also with the governmental and non-governmental humanitarian relief organizations that will be engaged for the long term. PMID- 12090200 TI - Enhancing the quality of humanitarian assistance: taking stock and future initiatives. AB - During the last five years, the debate on the performance of humanitarian assistance has intensified. The motivation to "do better" has come both from within the humanitarian agencies as well as from pressure exerted by the donors and the media. Paradoxically, until now, the voice of those who are to benefit from this assistance has not been heard. This paper is an overview of the most important initiatives to increase the quality of humanitarian assistance. The introduction of the logical framework and the increasing body of knowledge made available through guidelines have improved project management by measuring process and outcomes. Increasingly, evaluations are used to give account and to learn from experiences. But, current evaluation practice must develop in a wider variety of approaches more appropriate to create change of the operations in the field. Some agencies oppose new developments like the Sphere and the Humanitarian Accountability Projects, arguing that standards and regulation would undermine necessary flexibility to adjust responses to the local context, or be a threat to their independence. Nonetheless, standards are considered to be a prerequisite as reference to assess performance. Furthermore, it is hoped that a new breakthrough will be achieved by improved accountability towards beneficiaries. An option to address some of the gaps in the current quality assessment tools was to widen the perspective on performance from projects to the organisations behind them. Quality management models may provide the required framework, and they also can be used to embed current initiatives by organisations. Humanitarian organisations may want to develop forms of self-regulation rather than waiting for accreditation by donors. Another area in which progress is needed is a system wide approach to performance. At this level, the influence of political actors, donors, national governments, and other representatives of the parties in a conflict also should be assessed. It is their legal obligation to protect the basic right to assistance of persons affected by disasters, as enshrined in international law. PMID- 12090201 TI - Emerging issues and future needs in humanitarian assistance. AB - During the past two decades, there has been tremendous investment in the ability to intervene in disaster settings, and significant barriers remain to providing appropriate services to populations affected by natural and man-made calamities. Many of the barriers to providing effective assistance exist within the NGO community, and illustrate emerging needs for international agencies. These emerging needs include improving methods of recipient participation to promote the local health system, developing improved methods for quality assurance, enhancing options for personnel development, and addressing long-term needs of reconstruction and rehabilitation. Relief agencies face challenges on all levels to develop sound practices in providing humanitarian assistance that can lead to long-term benefits to populations affected by disaster. PMID- 12090202 TI - Integrating reproductive health into emergency response assessments and primary health care. AB - War-affected populations often are displaced for years. When primary health care is focused on the acute conditions that often present in the emergency phase of a complex emergency, insufficient attention often is directed towards other evolving needs of the population. Their reproductive health, psychosocial health, and problems with chronic diseases may be overlooked even after the situation stabilizes. This article examines currently available resources for conducting rapid assessments of health needs and services during complex emergencies. Their respective strengths and weaknesses are discussed, particularly for assessing a population's reproductive health needs, and for fostering the integration of reproductive health and primary health-care services, and for designing health services delivery. When more specific indicators are included in a needs assessment tool, the likelihood that the assessment results will influence the design and scope of the health program is increased. Needs assessments for primary health care that incorporate reproductive health indicators will assist health officials to integrate these services, and thus, use staff and facilities more efficiently, and will highlight areas of opportunity for providing services. PMID- 12090203 TI - Infant feeding practices in complex emergencies: a case study approach. AB - The majority of deaths associated with complex emergencies are attributed to infants and children under the age of five years. Most of these deaths are related to preventable diseases such as malnutrition, diarrhea, and malaria. Infant feeding emergencies have emerged as a major factor in complex emergencies. This paper reviews the current information relative to infant feeding, and uses four case studies as educational tools for the management of infant feeding emergencies. Child mortality rates in refugee population have been linked directly to protein-energy malnutrition (PEM). Breast feeding has many advantages over all other forms of feeding for children up to the age of two years of age. These advantages are discussed in detail in this paper. In addition, the appropriate and inappropriate uses of breast-milk substitutes (BMS) are discussed. Breast feeding also may play a role in the spread of HIV infections from the mother to the infant. However, in the setting of complex emergencies in the developing world, the risk of an infant dying of malnutrition and infection when not breastfed is likely to be greater than is the risk of death due to HIV acquisition through breastfeeding. The physiology of lactation is reviewed with particular reference to the roles of prolactin, oxytocin, and the feedback inhibitor of lactation (FIL) hormone. No medications have been demonstrated to augment milk production that can be used in a practical sense in complex emergencies. Lastly, the principles promulgated by the WHO and UNHCR for the feeding of infants and children in emergencies and for milk powder distribution are summarized. PMID- 12090204 TI - Physicians and international humanitarian law in complex emergencies: controversies and future opportunities. AB - This paper describes the areas in which the Geneva Conventions no longer are adequate as a source of legal description or prescription for the challenges faced by physicians working in complex emergencies. It covers the conceptual pitfalls facing the medical profession in connection with humanitarian interventions, which often are conventional military operations, but are not recognized as such because they may vary in some respects from more familiar forms of interstate conflict. Emerging categories of combatants who pose a major threat during complex emergencies also are identified. Opportunities to meet these challenges with the tools and culture of medicine are explored, and are proposed to the medical community as an opportunity for leadership. The paper proposes that new, epidemiological standards should be developed in order to identify the outbreak of armed conflicts and the trigger points for application of international humanitarian law. Such could replace the political model that presently underlies international humanitarian law. It also argues that international humanitarian law is not the starting point for application of humanitarian standards in war zones, but rather is built upon a peacetime medical culture that must be replicated in complex emergencies as a precursor to effective application of the law. PMID- 12090205 TI - Rehabilitating public health infrastructure in the post-conflict setting: epidemic prevention and preparedness in Kosovo. AB - The war in Kosovo in 1999 resulted in the displacement of up to 1.5 million persons from their homes. On the subsequent return of the refugees and internally displaced persons, one of the major challenges facing the local population and the international community, was the rehabilitation of Kosovo's public health infrastructure, which had sustained enormous damage as a result of the fighting. Of particular importance was the need to develop a system of epidemic prevention and preparedness. But no single agency had the resources or capacity to implement such a program. Therefore, a unique six-point model was developed as a collaboration between the Kosovo Institute of Public Health, the World Health Organization, and an international, non-governmental organization. Important components of the program included a major Kosovo-wide baseline health survey, the development of a province-wide public health surveillance system, rehabilitation of microbiology laboratories, and the development of a local capacity for epidemic response. While all program objectives were met, important lessons were learned concerning the planning, design, and implementation of such a project. This program represents a model that potentially could be replicated in other post-conflict or development settings. PMID- 12090206 TI - Communicating across cultures: improving translation to improve complex emergency program effectiveness. AB - Translation is a vital activity in Complex Emergencies (CEs) in which the responders and the affected populations do not share the same language or culture. This particularly applies to CEs in developing countries in which a lack of local resources usually results in the importation of foreign aid workers. This paper describes many of the common issues surrounding translation that can affect CE response effectiveness, issues that frequently are not appreciated by aid workers, including clinicians. The authors describe how these issues can arise, their effects, and outline approaches to addressing them. PMID- 12090207 TI - Preparing international relief workers for health care in the field: an evaluation of organizational practices. AB - The growth of the humanitarian aid industry has led to the proliferation of relief programs and the rapid rise in the number of relief personnel working in the field. One major necessity in developing successful international programs is appropriately trained field personnel. The purpose of this study was to evaluate the educational practice and training methods for field workers by non-government organizations (NGO). Of the 53 organizations surveyed, 64% responded that they sent health care workers to acute human emergencies. A majority of organizations, 31/53 (59%), used manuals as the primary method of training for workers before going into the field. Eighty-five percent of organizations (45/53) supplied their workers with trip briefings from prior personnel before going into the field, and 91% (48/53) had an on-site coordinator. Only 34% (18/53) provided classroom teaching or orientation prior to departure. The average number of months spent by workers abroad was < or = 1 for nearly half (49%) of the NGOs. Only 34% (18/53) of the NGOs required that personnel had previous international experience. Training of humanitarian workers varies significantly between non-governmental organizations. Lack of standardization in training programs and wide variation of provider preparedness indicates the need by NGOs for enhanced training for field personnel. PMID- 12090208 TI - Post-war Kosovo: Part 1. Assessment of prehospital emergency services. AB - The United Nations Mission in Kosovo (UNMIK) designated that the World Health Organization (WHO) develop health policy to assist in the recovery and rehabilitation of the post-war health system of Kosovo. As a critical part of the pre-policy evaluation, an assessment of current prehospital medical services was performed. This assessment identified a basic healthcare infrastructure upon which additional prehospital capabilities can be built, especially in communications, staffing, equipment, and transport services. To serve Kosovo properly in the future, it is recommended that capacity building must include the parallel development of emergency departments and specialty-trained physicians. PMID- 12090209 TI - Post-war Kosovo: Part 2. Assessment of emergency medicine leadership development strategy. AB - Since the return of the refugee population to Kosovo, attempts at development of an emergency medical system in Kosovo have met with varied success, and have been hampered by unforeseen barriers. These barriers have been exacerbated by the lack of detailed health system assessments. A multimodal approach of data collection and analysis was used to identify potential barriers, and determine the appropriate level of intervention for emergency medicine (EM) development in Kosovo. The four step, multi-modal, data collection tool utilized: 1) demographic and health systems data; 2) focus group discussions with health-care workers; 3) individual interviews with key individuals in EM development; and 4) Q-Analysis of the attitudes and opinions of EM leaders. Results indicated that Emergency Medicine in Kosovo is under-developed. This method of combined quantitative and qualitative analysis identified a number of developmental needs in the Kosovar health system. There has been little formal training, the EMS system lacks organization, equipment, and a reliable communication system, and centralized emergency centers, other than the center at Prishtina Hospital, are inadequate. Group discussions and interviews support the desire by Kosovar health-care workers to establish EM, and highlight a number of concerns. A Q-methodology analysis of the attitudes of potential leaders in the field, supported these concerns and identified two attitudinal groups with deeper insights into their opinions on the development of such a system. This study suggests that a multi modal assessment of health systems can provide important information about the need for emergency health system improvements in Kosovo. This methodology may serve as a model for future, system-wide assessments in post-conflict health system reconstruction. PMID- 12090210 TI - Post-war Kosovo: Part 3. Development and rehabilitation of emergency services. AB - The recent crisis in Kosovo led to nearly complete destruction of a healthcare system serving the needs of approximately 2 million people. Even prior to the crisis, the pre-existing healthcare system had inadequate provisions for the delivery of Emergency Medical Services. More than 440 diverse governmental and non-governmental organizations (NGOs) arrived to assist (and often compete) in the rehabilitation of Kosovo's healthcare needs. Each brought with them individual biases and strategies for how this rehabilitation should occur, and each faced numerous unforeseen barriers to the implementation of its programs. The authors used a four-step, multi-modal, needs assessment to gather information on the needs and potential barriers to the implementation of a program to rehabilitate emergency services as discussed in Part II. This paper chronicles the phases of the Emergency Medicine program development and the process of responding to barriers and changing needs. The program's successes and failures are noted, and the actual barriers encountered are reviewed. Overall, the needs assessment tool employed in this program was useful in the implementation of a program to restore and rehabilitate Emergency Services in Kosovo. The authors recommend the use of combined quantitative and qualitative methods for developing priorities for interventions in post-conflict settings following complex emergencies. PMID- 12090211 TI - Demographic methods to assess food insecurity: a North Korean case study. AB - In complex emergencies, especially those involving famine and/or widespread food insecurity, assessments of malnutrition are critical to understanding the population's health status and to assessing the effectiveness of relief interventions. Although the Democratic People's Republic of Korea (DPRK) has benefited from some of the largest, most sustained appeals in the history of the World Food Program (WFP), the government in Pyongyang has placed restrictions on international efforts to gather data on the health and nutritional status of the affected population. QUESTION: Lacking direct means to assess the nutritional status of the North Korean populace, what other methodologies could be employed to measure the public health impacts of chronic food shortage? The paper begins with a review of methods for assessing nutritional status, particularly in emergencies; a brief history of the North Korean food crisis (1995-2001), and a review of the available nutritional and health data on the DPRK. The main focus of the paper is on the results of a survey of 2,692 North Korean adult migrants in China. Recognizing certain biases and limitations, the study suggests that sample households have experienced an overall decline in food security, as evidenced by both the decline in government rations from an average of 120 grams per person per day to less than 60 grams per day, and by the increase in the percentage of households relying on foraging or bartering of assets as their principal source of food. It also is apparent that the period 1995-1998 has been marked by elevated household mortality, declining fertility, and steadily rising out-migration. Taken together, the signs point toward famine, whether that is defined as a discrete event--that is, as a regional failure in food production or distribution leading to elevated mortality from starvation and associated disease -or as a more complex social process whose sub-states include not only elevated mortality, but declining fertility, eating of alternative 'famine foods', transfer of assets, and the uprooting and separation of families. PMID- 12090212 TI - Complex emergencies in Indonesia. AB - Recently, Indonesia has experienced six major provincial, civil, armed conflicts. Underlying causes include the transmigration policy, sectarian disputes, the Asian economic crisis, fall of authoritarian rule, and a backlash against civil and military abuses. The public health impact involves the displacement nationwide of > 1.2 million persons. Violence in the Malukus, Timor, and Kalimantan has sparked the greatest population movements such that five provinces in Indonesia each now harbor > 100,000 internally displaced persons. With a background of government instability, hyperinflation, macroeconomic collapse, and elusive political solutions, these civil armed conflicts are ripe for persistence as complex emergencies. Indonesia has made substantial progress in domestic disaster management with the establishment of central administrative authority, strategic planning, and training programs. Nevertheless, the Indonesian experience reveals recurrent issues in international humanitarian health assistance. Clinical care remains complicated by absences of treatment protocols, inappropriate drug use, high procedural complication rates, and variable referral practices. Epidemiological surveillance remains complicated by unsettled clinical case definitions, non-standardized case management of diseases with epidemic potential, variable outbreak management protocols, and inadequate epidemiological analytic capacity. International donor support has been semi-selective, insufficient, and late. The militia murders of three UN staff in West Timor prompted the withdrawal of UN international staff from West Timor for nearly a year to date. Re-establishing rules of engagement for humanitarian health workers must address security, public health, and clinical threats. PMID- 12090214 TI - Controversies and guidelines in tissue-based HER-2/neu testing in breast cancer. AB - While it is tempting to think of the results of laboratory tests as quantitative, reproducible and accurate, many tests in fact have embedded variables that could lead to inconsistency and sometimes false reporting of results. Tissue-based testing of HER-2/neu in breast cancer represents an example of the latter. This article discusses the available technologies for HER-2/neu testing with some of the sources of testing errors. PMID- 12090213 TI - Principles of disaster management. Lesson 14. Criteria for assessing a program. AB - This lesson examines mechanisms that can be used for the evaluation of a program or project. The principal concern raised is whether the project has met its stated goals and objectives and whether the project has resulted in producing benefits to the affected society. Short-term (immediate) and long-term (developmental) contributions are discussed. The importance of projects contributing to increasing the absorbing capacity of the affected community for the next event is stressed. Twelve problems commonly encountered in program execution are defined. Optimal management attempts to identify potential pitfalls in advance, designing and implementing mechanisms to avoid them, and to deal with them if they should become manifest. Simply meeting the goals and objectives of the sponsoring organization is inadequate, as all responses must be coordinated and approved by the national coordinating agency. Thus, not only is the effectiveness of the project in meeting the defined goals and objectives important, but the project must be assessed in terms of the overall impact of the project on the society. Reference is made to using the structure provided by the Health Disaster Management: Guidelines for Evaluation and Research in the Utstein Style as promulgated by the Task Force on Quality Control of the World Association for Disaster and Emergency Medicine and the Nordic Society of Disaster Medicine. PMID- 12090215 TI - Medical laboratory industry--2007. Interview by C. Anne Pontius. PMID- 12090216 TI - Cardiac markers: cost-effective triage for MI admission and therapy. PMID- 12090217 TI - Incorporating HIPAA-relevant language into new vendor agreements. PMID- 12090218 TI - Latest changes to HIPAA still have problems; complying with OSHA needlestick directive. PMID- 12090219 TI - What "big picture" should inform professional chaplaincy? PMID- 12090220 TI - Pastoral counseling as a spiritual practice: an exercise in a theology of spirituality. AB - The author explores reasons for differentiating pastoral care from spiritual care and presents a theological exercise in articulating pastoral counseling as a spiritual practice. Viewing spirituality from systemic and constructivist perspectives, the author utilizes Paul Tillich's ontology of polarities. He describes spirituality as a subjective, "fuzzy," and inclusive concept and argues for the need for pastoral specialists to theologically articulate their ministry not only as a pastoral but also as a spiritual discipline. PMID- 12090221 TI - Uses of theories of depth psychology in ordained ministry and the institutional church. AB - Burgeoning understanding of unconscious processes should inform the work of the church and its ministries. Understanding derived from depth psychology can lead to deeper understandings of relationship with God and with one another, of the vicissitudes of spiritual life, of the meaning of ritual and sacraments, and to more profound and relevant scriptural hermeneutics and theological understandings. These claims are illustrated with examples. PMID- 12090223 TI - Rethinking supervision of ministry. AB - This article addresses the need for ongoing supervision in ministry and explores reasons for the avoidance of supervision. More specifically, twentieth century ideas regarding epistemology and Freud's "discovery" of the unconscious reveal important limitations in the commonly held model of supervision and at the same time provide reasons for the benefits of supervision. Further, this article proposes that individual unconscious motivations and religious institutional structures contribute to the avoidance of ongoing supervision in ministry. PMID- 12090222 TI - Developing an adequate "pneumatraumatology": understanding the spiritual impacts of traumatic injury. AB - Psychosocial interventions and systematic theology are primary resources for chaplains and congregational pastors who care for victims of physical trauma. Yet these resources may not be adequate to address the spiritual impacts of trauma. This article proposes a preliminary "pneumatraumatology," drawing on early Christian asceticism and Buddhist mysticism to describe one way of understanding the spiritual impacts of traumatic injury. It also suggests possible responses to these impacts informed by narrative/constructionist perspectives and Breggemann's understanding of the dimensions of spiritual transformation in the Hebrew Bible. PMID- 12090224 TI - Care teams and pastoral professionals: response to the health care changes of this decade. AB - Demographic changes already in motion will result in unprecedented population changes in the next four decades and beyond for which we have little time to prepare. In particular, as the population aged 65 and over grows, a significant cohort will manifest health-related needs of older age. Even in families with adequate health insurance, confronted with the chronic illness of loved ones, the need for continuous in-home support, including pastoral care, will become dire. For many families whose situations are complicated by low incomes, under insurance, and membership of minority populations, pastoral support will tax their congregations beyond present capabilities. Interfaith Care Partners' Care Team model of in-home pastoral support is the most effective means of delivering the pastoral support such families need. It calls for a creative linking of hospital out-patient services with individuals' congregations, and, particularly, of a stronger connection between hospital chaplaincy services and community clergy. PMID- 12090225 TI - Worship with a twist. AB - The core values of hospitality, respect, and innovation help to guide and initiate changes in the worship services of a rehabilitation hospital. The Antiphon of the day, special hymnals containing residents choices, and inclusion in the prayers of the day helped achieve these goals. Residents are constantly given choices with the Chaplain ready to keep the service moving smoothly. In these ways the core values are included in all services. Specific resources are listed, and the article is followed by a response from another chaplain with a somewhat different approach. PMID- 12090226 TI - The road less traveled: thoughts on choosing a seminary. PMID- 12090227 TI - Ordinary things ... of enormous value! PMID- 12090228 TI - Serving well. PMID- 12090229 TI - Blessed watchfulness. PMID- 12090230 TI - Proceedings of the 2nd Sino-European Congress of Laboratory Medicine. Shanghai, China. April 25-27, 2001. PMID- 12090231 TI - Public ambivalent over privacy issues in healthcare. PMID- 12090232 TI - Gathering the big picture: Healthcare Issues Survey, year three. PMID- 12090233 TI - Profile: Dr. Alan Hudson. Interview by Cynthia Martin. AB - Besides enviable success as a healthcare leader, primarily as President and CEO for 10 years at the University Health Network in Toronto, Dr. Alan R. Hudson is a neurosurgeon whose many awards and professional activities have helped to establish him as a world-class research. PMID- 12090234 TI - More on nursing workplaces. PMID- 12090235 TI - Retaining digital images. PMID- 12090236 TI - Evaluating the effect of stage-specific survivorship on the N(e)/N ratio. AB - Evaluating effective population size (Ne) and the effective size to census size ratio (Ne/N) in species with Type III survivorship curves is complicated when key demographic parameters [mean (k macro) and variance (V(k)) of family size] are measured during early life stages. The method of Crow & Morton (1955) for scaling demographic data collected at a juvenile stage to expected values at adulthood is extended to consider sequential episodes of random and family correlated survival. Results show the following: (i) The order in which the episodes of random and family-correlated survival occur does not affect N(e) or N(e)/N; (ii) If a population experiences an episode of family-correlated survival, N(e)/N scaled to its expected value in a population of constant size (k macro= 2) is simply the survival rate during the family-correlated stage. If multiple such stages occur, scaled N(e)/N is the product of the survivals during all family correlated life stages; (iii) Under the assumption of random post-enumeration survival, adjusting the variance effective size to its expected value at k macro= 2 is equivalent to computing the inbreeding effective size at the earlier life stage. Application to experimental data for hatchery populations of Pacific salmon (Oncorhynchus spp.) indicates that nonrandom survival during the marine phase led to estimated reductions in effective size of 0-62 (mean 19) in 12 different cohorts. This approach can provide insights into N(e)/N in highly fecund species, including some marine species in which N(e) has been estimated to be several orders of magnitude less than N. PMID- 12090238 TI - Academic health sciences centres: not enough has changed. PMID- 12090237 TI - An example of successful partnering. PMID- 12090239 TI - Academic health sciences centres: several perspectives missed. PMID- 12090240 TI - Decline in perceptions of quality of care may be ending. PMID- 12090241 TI - The specificity of the yeast and human class I ER alpha 1,2-mannosidases involved in ER quality control is not as strict previously reported. PMID- 12090242 TI - AMA unit says 'no' to mandatory HIV testing. PMID- 12090243 TI - Garlic supplements can impede HIV medication. PMID- 12090244 TI - What happens to T-cells explained. PMID- 12090245 TI - Re: utility of nonselective abdominal aortography in demonstrating ovarian artery collaterals in patients undergoing uterine artery embolization for fibroids. PMID- 12090246 TI - Acromegaly: a new therapy. AB - BACKGROUND: The treatment of acromegaly can be challenging. Despite a multimodality approach (surgery,radiation, dopamine agonists, somatostatin analogs), many patients do not achieve normalization of serum insulin-like growth factor I (IGF-I) concentrations. METHODS: The author discusses the characteristics and indications of pegvisomant therapy for patients with acromegaly and compares the use of this newly developed GH receptor antagonist with other pharmacological agents such as somatostatin and dopamine agonists. RESULTS: Therapy with pegvisomant allows serum IGF-I concentrations to be normalized in up to 97% of patients with acromegaly, including those who have failed other treatment modalities. With this agent,circulating GH levels increase as a result of the drop in IGF-I levels. The rise is rapid (within 2 weeks) and does not appear to be progressive over time. CONCLUSIONS: Published studies have shown pegvisomant to have efficacy in the treatment of acromegaly. As it appears to be well tolerated and safe, this novel compound may be an important therapeutic option for patients with acromegaly. Additional study of this novel agent and its mode of action is warranted. PMID- 12090248 TI - Yeast Whi2 and Psr1-phosphatase form a complex and regulate STRE-mediated gene expression. AB - BACKGROUND: In response to various stressful situations, including diauxic conditions, the Msn2 and Msn4 transcription factors induce STRE-mediated gene expression of many stress responsive genes in Saccharomyces cerevisiae. This is called the general stress response. The whi2 cells in the stationary phase are smaller than wild-type cells. RESULTS: Here we demonstrate that STRE-mediated gene expression in whi2 cells is reduced to half of that in the wild-type cells under various stress conditions. It is also delayed for several hours when the mutant cells enter the stationary phase. Using the two-hybrid system, we isolated a WHI2-interacting gene, PSR1, which is one of the redundant genes encoding plasma membrane phosphatases. whi2 and psr1 psr2 mutants had similar phenotypes, including reduced STRE-mediated gene expression, higher sensitivity to sodium ions and heat shock, and hyper-phosphorylation of Msn2. The phosphatase activity of Psr1 was necessary for the full activation of STRE-mediated gene expression. Furthermore, both Psr1 and Msn2 were co-immunoprecipitated with Whi2. CONCLUSIONS: Thus, Whi2 and its binding partner, Psr1-phosphatase, are required for a full activation of the general stress response, possibly through the dephosphorylation of Msn2. These results may explain why stationary phase whi2 cells are small. PMID- 12090249 TI - Cellular reprogramming in the F3 mouse with paternal F0 radiation history. AB - PURPOSE: In the mouse, paternal F0 acute irradiation of Type B spermatogonia produces biological effects in offspring, including altered signalling kinase activities and protein levels. It was hypothesized that these effects represented cellular reprogramming that would alter the response of somatic cells in these offspring to an acute ionizing radiation exposure. MATERIALS AND METHODS: Nineteen-day-old third generation (F3) CD1 mice with and without an acute 1.0 Gy paternal F0 radiation history each received an acute dose of 1.0 Gy from attenuated 137C n-rays. Kidney PKC and MAPK activities, and p53 protein levels were evaluated immediately following F3 irradiation. The same endpoints and DNA damage were evaluated in kidney-derived fibroblast primary cell cultures 3 weeks post-irradiation. RESULTS: Kidneys had significantly decreased PKC and MAPK activities and p53 protein levels related to F0 irradiation and increased PKC and MAPK activities following F3 irradiation irrespective of F0 radiation history. Kidney-derived fibroblasts had significant changes or strong trends for all selected endpoints based upon cross-interaction of F0 radiation history with F3 irradiation. Comet assays demonstrated significantly increased DNA damage in fibroblasts related to F0 irradiation and increased DNA damage following F3 irradiation. However, significantly decreased F3 irradiation damage was demonstrated based upon cross-interaction of F0 radiation. CONCLUSIONS: The data suggest that irradiation of paternal F0 Type B spermatogonia resulted in cellular reprogramming causing offspring with this radiation history to have altered responses to acute somatic n-irradiation. PMID- 12090251 TI - The three-legged stool. PMID- 12090250 TI - Adjusting to change: linking family structure transitions with parenting and boys' adjustment. AB - This study examined links between family structure transitions and children's academic, behavioral, and emotional outcomes in a sample of 238 divorcing mothers and their sons in Grades 1-3. Multiple methods and agents were used in assessing family process variables and child outcomes. Findings suggest that greater accumulations of family transitions were associated with poorer academic functioning, greater acting-out behavior, and worse emotional adjustment for boys. However, in all three cases, these relationships were mediated by parenting practices: Parental academic skill encouragement mediated the relationship between transitions and academic functioning, and a factor of more general effective parenting practices mediated the relationships between transitions and acting out and emotional adjustment. PMID- 12090252 TI - Patient safety. One giant Leap? PMID- 12090253 TI - Above and beyond? PMID- 12090254 TI - Lessons learned as a surgical chairman. PMID- 12090255 TI - Solid organ transplantation in HIV-infected individuals. PMID- 12090256 TI - Remediation of ammonia-rich minewater in constructed wetlands. AB - A three-year study of ammonia removal from minewater was carried out employing constructed wetland systems (surface flow wetland and subsurface flow wetland cells) at the former Woodey Mine in West Yorkshire, UK. The 1.4 Ha surface flow wetland (constructed in 1995) reduced the ammonia concentration from 3.5 - 45 mg l(-1) to < 2.3 mg l(-1) during the first half of the study and to essentially zero in the last year (2000 - 2001). About 25% of contained ammonia was converted to nitrate, about 10% was consumed by the plants and up to 30% was converted to nitrogen gas. This maturation effect was attributed to increased depth of sludge from sedimentation of ochre, providing increased surface area for immobilisation of ammonia oxidising bacteria. The surface flow wetland finally removed 2.3 g m( 2) day(-1) ammonia in comparison with 3.8 g m(-2) day(-1) for the subsurface flow (pea gravel) wetland cells, constructed for the present work and dosed with ammonium salts. Removal of ammonia by both systems was consistent with well established mechanisms of nitrification and denitrification. It was also consistent with ammonia removal in wastewater wetland systems, although the greater aeration in the minewater systems obviated the need for special aeration cycles. The general role of wetland plants in such aerated conditions was attributed to maintaining hydraulic conditions (such as hydraulic efficiency and hydraulic resistance of substratum in subsurface flow systems) in the wetlands and providing a suspended solids filter for minewater. PMID- 12090257 TI - [Lead and zinc recovery by adsorption on peat moss during municipal incinerator used lime decontamination]. AB - The objective of this research was to evaluate the efficiency of peat moss for lead and Zn recovery from alkaline leachates (pH 11.5) produced during decontamination of municipal incinerator fly ash. Tests carried out with peat moss columns (density of 0.13 g m l(-1)) gave very high removal yields for lead (98.9 to 100%) and Zn (98.4 to 99.8 %). The initial metal concentrations in the leachates were 126 to 138 mg Pb l(-1) and 14.4 to 23.5 mg Zn l(-1). The columns were fed using two rates (20 and 40 ml min(-1)), which correspond respectively to 17.6 and 8.8 min as contact time on peat moss. Adsorption efficiencies of 16 to 18 mg Pb g(-1) and 1.7 to 3.2 mg Zn g(-1) have been measured during this fly ash leachate treatment study. The adsorbed toxic metals can be desorbed using a chlorhydric acid solution. The peat moss can be regenerated and reused for several adsorption cycles without loss of the lead and Zn adsorption efficiencies. PMID- 12090258 TI - Ozonation of textile wastewater: physicochemical and phytotoxic aspects. AB - The purpose of this work was to evaluate the performance of ozonation as a technique to treat textile effluents. This performance evaluation was made using physico-chemical parameters and phytotoxic endpoints (i.e. biomass growth enzyme activities - catalase and peroxidase). After ozonation, the color absorbance (523 nm) was reduced by 80.9% and the pH decreased from 10.9 to approximately 7.5 while COD reduction reached nearly 87%. Phytotoxicity tests carried out in solution with three plant species (i.e. soybean, rice and wheat) allowed us to compare toxicity data of both, raw and ozonated effluents. The biomass (fresh weight) of the 3 plant species clearly decreases as a function of the increase in raw effluent concentration with LOEC values of 100% for rice and wheat, and 50% for soybean. Activities of catalase and peroxidase in the 3 plant species exposed to the raw textile effluent were significantly different from plant controls. The LOEC values for catalase were 6.2% (in wheat and soybean plants), and 25% (in rice plants), while for peroxidase the LOEC values were 6.2% (in rice plants), 25% (in soybean plants), and 50% (in wheat plants). Reduction in the phytotoxicity of textile effluent samples after ozonation is likely due to the elimination of the toxic organic fraction oxidized during this process. Regarding endpoint sensitivity, our results showed that biomass endpoint was less sensitive than biochemical endpoint. In conclusion, ozonation was relatively effective in reducing physico-chemical parameters and phytotoxic effects of textile effluents. Plant enzyme assays used in this study proved to be a sensitive tool in order to determine the toxicity of industrial effluents and may be used to evaluate efficiency of established (or emergent) wastewater treatment technology. PMID- 12090259 TI - [44th annual meeting of the Japan Geriatrics Society. Tokyo, Japan. June 12-14, 2002. Abstracts]. PMID- 12090260 TI - Retreatment in endodontics: treatment decisions by general practitioners and dental teachers in Finland. AB - OBJECTIVE: To evaluate treatment decisions of general practitioners (GPs) and dental teachers (DTs) in cases of endodontically treated teeth and to test the hypothesis that dentists' treatment decisions are independent of their working sector and other practice characteristics. DESIGN: A questionnaire based on four endodontic cases was mailed to 400 GPs and 47 DTs. The GPs were selected by stratified randomisation by gender, and main occupation (public and private sector). Others were all full-time DTs, representing clinical disciplines other than surgery and orthodontics. The endodontic cases were described in detail: one with a radiograph enclosed, three with statements on radiographic examinations. For each case, respondents chose the optimal treatment from alternatives, later reclassified as 'no therapy now' (further examination within 3-12 months), and 'active treatment' (immediate retreatment or periapical surgery). RESULTS: For Case 1 (tooth 25, asymptomatic, in normal occlusion, the radiograph revealed an underfilled root canal and a crown with a post, widened periodontal ligament space around the apex but no radiolucency), active treatment was chosen more frequently by private than by public dentists (11% vs. 4%, P=0.01), but neither sector differed (P>0.05) from DTs (6%). Decisions in favour of active treatment were associated with higher share of adult patients (OR=2.8; 95%CI: 0.3, 1.0; P=0.01). Cases 2-4 showed no difference in treatment decisions by practice characteristics. CONCLUSION: Treatment decisions by GPs and DTs were quite similar for each case and in most cases independent of the dentist's work and practice related characteristics. PMID- 12090261 TI - Incidence and distribution of root canal treatment in the dentition among a Jordanian sub population. AB - AIM: To determine and compare the incidence of endodontically treated teeth in the maxilla and mandible, and to present the frequency of endodontic treatment for each tooth in the dentition in a group of the Jordanian population; then to compare these results with previously published surveys. METHOD: A total of 1,404 root canal treated teeth were examined clinically and radiographically. RESULTS: Most teeth were treated in a one visit procedure (40.1%). The results showed that 77.7% of the treated teeth were in the maxilla and 22.3% were in the mandible. Posterior teeth accounted for 39.0% of all treated teeth, among which, 20.3% were molars. The most frequently treated tooth was the maxillary central incisor tooth (39.6%), with the maxillary lateral incisor second (10.1%). Furthermore, the maxillary incisors alone accounted for almost half (49.7%) of the cases. The maxillary first molar was the third most frequently treated tooth (8.2%) with the mandibular first molar at 7.5%. The lowest frequency of treatment was for mandibular lateral incisors and canines which made up only 1.1% and 1.9% respectively. CONCLUSION: Comparison with previously published surveys revealed a lower percentage of molar endodontics than in other surveys. PMID- 12090262 TI - Oral health related knowledge, behaviours, attitude and self-assessed status of primary school teachers in Tanzania. AB - AIM: To assess the oral health related knowledge, attitudes, behaviours and self assessed status of primary school teachers in Rungwe, Tanzania. SUBJECTS: 195 final year teacher trainees and 239 in-service teachers. METHOD: A structured questionnaire. RESULTS: School teachers were generally well informed and had moderate attitudes and behaviours to oral health related issues. However, a few but important deficiencies in this regard have been identified on which recommendations are presented. A substantial proportion of teachers reported having dental problems. However, the teachers had positive attitudes towards oral health education to pupils as part of a teaching curriculum. A professional support is called upon for teachers from the oral health personnel in terms of oral health education training at teachers' colleges, to prepare the teacher trainees for their future task. CONCLUSION: In-service teachers need to be motivated to improve their awareness on sound oral health information, attitudes and behaviours. This will facilitate their role as school oral health education providers in Tanzanian primary schools. PMID- 12090263 TI - Effectiveness of primary school-based oral health education in West Java, Indonesia. AB - A study in West Java has indicated that involvement of primary health care personnel and schoolteachers in oral health education (OHE) at primary schools is a feasible approach that is sustainable. AIM: The present study aims to assess the effects of that school-based OHE programme on pupils who had completed the programme one and a half years ago. METHODS: Eight experimental and six control primary schools in the same area participated in the study. Out of each school 10 children, aged 8-12-years old, were randomly selected. RESULTS: ANOVA with age and gender as co-variables showed statistically significantly lower (21%) habitual plaque scores among children from experimental schools as compared to those from control schools. Tooth brushing effectiveness had significantly improved among experimental children and they took longer for tooth brushing when supervised. The dmft/dmfs values were comparable but the DMFT/DMFS values of children from experimental schools tended to be lower (not statistically significant) than of those from control schools. Differences in oral health knowledge were apparent but self-reported habits pertaining to oral health were comparable between children from experimental and control schools. CONCLUSION: This school-based OHE programme had a moderate positive effect on oral health knowledge and on habitual plaque levels and on the effectiveness of tooth brushing. The effects on caries levels and on self-reported behaviour were inconclusive. PMID- 12090264 TI - Utilisation of dental services among a Turkish population in Witten, Germany. AB - AIM: To describe the oral health status and the dental service use pattern of a Turkish population in Witten, Germany, and to assess the factors affecting this use pattern. DESIGN: Cross-sectional. PARTICIPANTS: 532 Turkish people older than 12 years. METHODS: The data were obtained through a structured questionnaire and oral examination. The World Health Organisation methodology was followed in assessing the oral health status. RESULTS: Comparison of the study population with German studies showed a similar caries experience in younger groups, but in older groups a lower treatment level. The proportion of regular dental attendees indicated no difference according to age. A person was more likely to use dental services regularly if he had a better level of German language and preferred German services. Perceived oral condition, educational level and sex were also important for care pattern. Regular use of services was negatively associated with the mean number of decayed, missing and periodontally involved teeth, but positively related to the number of filled teeth. CONCLUSIONS: Since factors reflecting the level of adaptation to Germany were also important determinants of the dental utilisation pattern, efforts to improve the utilisation of services should be based on the socio-educational development of Turkish people in Germany. PMID- 12090265 TI - Self reported use of dental services among employed adults in Sri Lanka. AB - AIM: To determine the patterns and variables associated with self-reported use of dental services among employed adults in Sri Lanka. DESIGN: A cross sectional study using a self-administered questionnaire. SETTING: Five representative public sector institutions in the city of Kandy, Sri Lanka. PARTICIPANTS: A total of 210 employees. RESULTS: Ninety per cent of the employees had used dental services, the majority for symptomatic reasons. Of the respondents, 63% had visited a dentist in the preceding 24 months. According to the logistic regression analysis, gender and presence of pain within the past six months were significant predictors of dental utilisation. CONCLUSIONS: Andersen and Newman's behavioural model of health services utilisation was of limited value in explaining dental utilisation of this sample of employed adults as only one predisposing factor and a single factor related to need were significant predictors of the use of dental services. PMID- 12090266 TI - Comparison of United States and Korean dental hygiene students using the Hiroshima university-dental behavioural inventory(HU-DBI). AB - OBJECTIVE: To compare cross-national differences of dental health behaviour among dental hygiene students. DESIGN: Cross-cultural differences. SETTING: United States (USA) and Korea. SUBJECTS AND METHODS: One hundred and twenty-six dental hygiene students in USA and 246 in Korea were surveyed using the Hiroshima University-Dental Behavioural Inventory (HU-DBI) (in English and Korean versions respectively). RESULTS: There were considerable differences in oral health behaviour between USA and Korean dental hygiene students. Of great significance were findings that while only a small proportion of the USA students (1%) reported gum bleeding when they brushed their teeth, 37% of the Korean students did so (P<0.001). Furthermore, only 19% of the Korean students had been told by their dentist that they were performing a high level of plaque control, contrasted with 76% of the USA students. Logistic regression model showed that it was possible to distinguish USA students from Korean peers with a probability of more than 90% by using the HU-DBI. CONCLUSIONS: There were considerable differences in dental health attitudes/behaviour among dental hygiene students in the two countries. The variation in the favourable attitudes/behaviour toward oral health appeared to reflect the students' clinical training experience in Korea. PMID- 12090267 TI - Oral aspects of Crohn's disease. AB - Crohn's disease is a chronic inflammatory disease, which may involve any part of the gastrointestinal tract, including the oral cavity. This review gives an overview of the oral findings observed in patients with Crohn's disease and the potential implications of the disease for dental management are discussed. PMID- 12090268 TI - Effects of light on gravitaxis and velocity in Chlamydomonas reinhardtii. AB - The effects of light on gravitaxis and velocity in the bi-flagellated green alga Chlamydomonas reinhardtii were investigated using a real time automatic tracking system. Three distinct light effects on gravitaxis and velocity with parallel kinetics were found. Photosynthetically active continuous red light reversibly enhances the swimming velocity and increases or decreases the precision of gravitaxis, depending on its initial level. Blue light flashes induce fast transient increases in velocity immediately after the photophobic response, and transiently decrease or even reverse negative gravitaxis. The calcium dependence of this response, its fluence-response curve and its spectral characteristics strongly suggest the participation of chlamy-rhodopsin in this effect. The third response, a prolonged activation of velocity and gravitaxis, is also induced by blue light flashes, which can be observed even in calcium-free medium. PMID- 12090269 TI - Repetition of the classical Boysen-Jensen and Nielsen's experiment on phototropism of oat coleoptiles. AB - The classical experiment of phototropic response as reported by Boysen-Jensen and Nielsen (1926), which supports the Cholodny-Went theory, was repeated in detail. In the original experiment, etiolated oat (Avena sativa L. cv. Victory) coleoptiles with mica inserted into their tip only showed a positive response when the mica was placed parallel toward the light source and not if it was inserted perpendicularly. On the contrary, we found a positive response irrespective of whether the mica was inserted parallel or perpendicularly to the light source. Damage owing to rude splitting severely reduced the response upon perpendicular insertion. These results invalidate the Boysen-Jensen and Nielsen's experiment as a support of the Cholodny-Went theory and lend support to the Bruinsma-Hasegawa theory ascribing phototropism to the local light-induced accumulation of growth inhibitors against a background of even auxin distribution, the diffusion of auxin being unaffected. PMID- 12090270 TI - Virtual presence: one step beyond reality. PMID- 12090271 TI - Yucca Mountain: should we delay? PMID- 12090273 TI - Yucca Mountain: should we delay? PMID- 12090272 TI - Yucca Mountain: should we delay? PMID- 12090274 TI - HIV-1 diversity and vaccine development. PMID- 12090275 TI - Getting smaller and smaller. PMID- 12090276 TI - Removal dynamics of airborne road dust in a ventilated airspace. AB - We derive a simple linear dynamic equation to describe the removal mechanisms of airborne road dust from a ventilated airspace. The dynamic equation is sufficiently to take into account the simultaneous removal effects of turbulent coagulation, turbulent diffusive deposition, gravitational sedimentation, and airflow pattern within a ventilated airspace. Three dimensionless parameters TC, TD, and GS that characterize the relative effects of turbulent coagulation, turbulent diffusive deposition and gravitational settling, respectively, in a ventilated airspace were introduced to generalize the removal dynamics of airborne road dust. An environmental chamber test was carried out not only to determine the particle size distributions but also to verify the removal dynamics of airborne road dust in a ventilated airspace. Our results demonstrate that there is no significant variation for particle size distributions of road dust obtained from urban and suburban areas in north Taiwan region and both followed a lognormal distribution with average geometric mean diameter of 1.08 +/- 0.02 microm and geometric standard deviation of 2.59+/-0.03. Measured values match the simulated values with an r2 value of 0.93, whereas the overall RMSE value of 2.36 +/- 1.05 mg m(-3) is low, indicating that the ability to predict the removal dynamics of airborne road dust within a ventilated airspace using an average particle size based linear equation. Effects of TC, TD, GS, and various ventilation systems on the time-dependent road dust concentrations are also justified. PMID- 12090277 TI - An evaluation of residues at an oil refinery site following fires. AB - Soil pollution at the oil refinery at Novi Sad following destruction of crude oil and its products in storage tanks during the Kosovo conflict was investigated. More than 100,000 t of crude oil and its products were destroyed, and about 90% of these were burnt off, 10% leached and 130 t recovered. The acute injection of polycyclic aromatic hydrocarbons (PAHs) to the air of the town was widespread depending on the weather conditions and ranged from 1-431,000 ng/m3. The presence of PAHs, polychlorinated biphenyls (PCBs), and elements in the surface zone and soil core samples taken from various sites were determined up to a depth of 100 cm. Concentrations of PAHs were in the range between 0.75 and 86.19 microg/g dry soil. The contaminated soil can be expected to act as a permanent pollution source, while the mobile constituents are likely to cause groundwater pollution. PMID- 12090278 TI - Water infiltration through diesel-contaminated soil. AB - The water infiltration characteristics through soil contaminated by diesel was investigated. Columns were packed with soil containing 15, 10, 5, 1, and 0 percent diesel on mass basis. Two soil types were used, namely, clay loam and sand. The results showed that the water infiltration rate through clay loam soil containing 1 percent diesel was about 50 percent of its value through diesel free soil. This relative rate dropped to less than 35 and 25 percent when soil contaminated with 5 and 10 percent diesel was used, respectively. Increasing the initial diesel concentration beyond 10 percent does not cause further significant drop in the water infiltration rate. Similar trend was observed when the sandy soil was used. However, the effect of initial diesel concentration on the water infiltration rate was less significant compared to the results obtained using the clay loam soil. The relative infiltration rates through the sandy soil ranged from about 80 percent in the sand containing 1 percent diesel to less than 35 percent in sand containing 15 percent diesel. The variation of the relative infiltration rate with degree of diesel saturation does not appear to be linear. Post experiment dissection of the columns showed that water partially displaced diesel pushing it down to lower soil layers. The percentage of diesel displaced by water in the sandy soil was higher than that in the clay loam soil. PMID- 12090279 TI - Influence of alcohol cosurfactants on surfactant-enhanced flushing of diesel contaminated soil. AB - The effects of medium-chain alcohols on the surfactant-enhanced diesel removal from contaminated sandy soil were investigated. The mixture of Tween 80, which has a double bond in its hydrophobic structure, and hexanol showed the best performance of diesel removal among the combinations of surfactants (Tween 20, Tween 60, and Tween 80) and alcohols (n-butanol, n-pentanol, and n-hexanol) used in this study. The efficiency of diesel flushing increased as the alkyl-chain length of the alcohol and the alcohol fraction in the flushing solution increased. Meanwhile, the removal rate was reduced when the surfactant concentration was too high most likely due to the surfactant aggregation or emulsion formation. A pulsed pumping of flushing solution was not beneficial for improving the removal rate under the experimental conditions described in this study. PMID- 12090280 TI - Heavy metal adsorption by crude coniferous barks: a modelling study. AB - Equilibria and mechanisms involved in the adsorption process of metal ions (Cr3+, Cu2+, Ni2+, Pb2+ and Zn2+) from aqueous solutions using coniferous barks as biosorbent substrate were investigated. Crude barks were used in this study since previous experiments showed a decreasing uptake for chemically treated barks in the considered granulometry. In our experimental conditions, the maximum binding capacity of barks followed the decreasing order Cr3+ > Cu2+ > Pb2+ > Ni2+ > Zn2+ whereas their general binding affinity decreased as: Pb2+ > Cr3+ > Ni2+ > Zn2+ > Cu2+. Adsorption isotherms at the optimal physicochemical conditions were established and the adsorption phenomenon was described by the non-competitive Langmuir adsorption model which fitted well the experimental data. An evaluation of adsorption capability was carried out using model parameters which were graphically determined. Models for removal of cations have been discussed; they represent efficient tools for predicting the behaviour of the biosorbents in metal ion adsorption systems. PMID- 12090281 TI - Influence of thermal regime of soil on the sulfur (S) and selenium (Se) concentration in potato plants. AB - Three consecutive years of field experiments were carried out to investigate the effect of different root temperatures, induced by the application of mulches on the concentration of sulfur (S) forms (organic-S, total-S and SO4(2-)) and Se in different organs of potato plants (roots, tubers, stems and leaves). Four different plastic covers were used (T1: transparent polyethylene; T2: white polyethylene; T3: white and black coextruded polyethylene, and T4: black polyethylene), using uncovered soil as control (T0). The different treatments had a significant effect on mean root temperatures (T0 = 16 degrees C, T1 = 20 degrees C, T2 = 23 degrees C, T3 = 27 degrees C and T4 = 30 degrees C) and induced a significantly different response in the S forms and Se concentration, showing the T3 treatment (27 degrees C) the greatest concentration of total S and organic S in the stems and leaflets. The Se reached higher levels in the roots and tubers in T3. With regard to possibilities in phytoremediation, it is necessary to control the thermal regime of the soil to optimize the accumulation of elements. PMID- 12090283 TI - The benefits and risks of using river sediment for Vietnamese agriculture: a case study of the Nhieu Loc canal in Ho Chi Minh City. AB - This study was conducted to assess the potential of river sediment for amendment of agricultural land with respect to heavy metal mobilization. The mobility and concentration of bioavailable heavy metal containing in the sediment were evaluated with the use of DTPA (diethylenetriamine pentaacetic acid) extraction techniques. Sum of metals in exchangeable and carbonate fractions obtained from sequential extraction scheme was used to evaluate the potential plant available. Four land blocks of size (1 m x 5.8 m) were prepared for cabbage field experiment and applied with different loading rates of sediment (approximately 28 and 42 ton/ha or equivalent to 100 kgN/ha and 150 kgN/ha). In parallel test, land blocks treated with fertilizer at similar nitrogen loading rates were also conducted for comparisons. Pure soil plot was used as a control system. Results show that both plant available and total heavy metal concentration were found higher in those plots treated with higher loading rates of sediment (42 ton/ha). The concentrations of heavy metals (Cu,Zn,Pb) in soil were decreased with time after crop harvesting and the residues remain in the soil were well below those of the UK, USA and Germany standards. Simple linear regression was used to show the correlation between the DTPA extractable metals and the plant metals fractions (root and shoot) resulting from fertilizer and sediment applications in the amended soils. In most cases, plant available metal concentrations showed positively correlated with plant metal uptake concentrations (R2 = 0.01-0.95, n = 3). Concentration of metals in the cabbage tissue detected: Zn at 131.53+/-10.43 mg kg(-1) DM, Cu at 20.84+/-1.62 mg kg(-1) DM, Pb at 0.09+/-0.01 mg kg(-1) DM, Cd at 0.21+/-0.09 mg kg(-1) DM and Ni at 0.44+/-0.05 mg kg(-1) DM. None of these metals concentration exceeded the tolerable limits of plant. One-way analysis of variance (ANOVA) was applied to find out the effect of different soil amendments on cabbage yield in term of fresh and dry weights. The sequence of crop yields in this field studies for amended soils is chemical fertilizer > sediment > control soil. PMID- 12090282 TI - Effect of agitation and aeration on bioconversion of domestic wastewater sludge in a batch fermenter. AB - Effects of agitation and aeration rate on microbial treatment of domestic wastewater sludge were investigated in a batch fermenter using mixed culture of Penicillium corylophilum and Aspergillus niger. It was found that liquid state bioconversion (LSB) of wastewater sludge was highly influenced by the effects of agitation and aeration. The maximum production of sludge cake and reduction of organic substances in treated sludge were recorded at 150-200 rpm of agitation speed and 0.5 vvm of aeration rate after 72 h of treatment. No effective results were observed at higher rate of agitation (300 rpm) and aeration (1.5 vvm) as compared to optimum values. The results showed that the minimum level of air saturation (pO2) was adequate to maintain the bioconversion process. PMID- 12090284 TI - Effects of high temperature melting on fractionation of heavy metals in sewage sludge. AB - Sewage sludge from aerobic treatment plant was found to contain high amounts of heavy metals. Research was carried out to investigate the speciation and leaching behavior of heavy metals when using high temperature melting technology for treatment. This was achieved by conducting a sequential chemical extraction procedure and EP-TOX leaching test. The thermal treatment led to increased shift of metals from organic fraction to residual fraction, indicating that the thermal treatment caused metals in sewage sludge to become stable. Furthermore, results from leaching test revealed that metals were not leached from the final product after thermal treatment and this was verified using US EPA standard limits. Results from this study indicated that melting technology could convert the sludge to product that can be either reused or landfilled without an adverse environmental impact. PMID- 12090285 TI - Biodegradation and adsorption of phenol using activated carbon immobilized with Pseudomonas putida. AB - This paper examined the removal efficiency of phenol from aqueous solution using a suspended culture of Pseudomonas putida (ATCC 3180) or the activated carbon on which the microorganism was immobilized. The kinetics of phenol degradation by immobilized and pure cells was studied. Experiments were performed at various phenol concentrations (0.1-0.4 g/L), pH, temperature (30-36 degrees C), and concentrations of glucose (0.5-0.7 g/L) and (NH4)2SO4 (0.5-0.7 g/L). The presence of activated carbon markedly enhanced the degradation efficiency, showing its ability of protecting microbes from confronting shock loads of organic pollutants. Degradation rate increased with increasing substrate concentration and decreased after reaching a maximum, indicating substrate-inhibition kinetics. In addition, the degradation rate for immobilized cells was much higher than that of free cells. The inhibition effect for phenol degradation was described by the Andrews model. The kinetic parameters were also determined. PMID- 12090286 TI - Effect of catalyst preparation conditions on the hydrodesulfurization of thiophene over Co-Mo/gamma-Al2O3. AB - The purpose of this research was to study the effects of preparation conditions on the catalytic properties of the Co-Mo/gamma-Al2O3 catalyst. The work included catalyst preparations and reactions. In the preparations, cobalt-impregnated Mo/gamma-Al2O3 (designated as IcIM) was found to have a promoting effect on the hydrodesulfurization (HDS) of thiophene. Activity and stability of IcIM was higher than that of Mo/gamma-Al2O3. Conversely, when cobalt was added onto Mo/gamma-Al2O3 by the mechanical mixing method, no promoting effect was observed. Mo/gamma-Al2O3 was also prepared using the two different methods (incipient impregnation or mechanical mixing). The differently prepared Mo/gamma-Al203 resulted in no obvious difference in activity of IcIM. It was further found that Co-Mo/gamma-Al2O3 activity initially increased appreciably with Mo content and leveled off at Mo contents above 9 wt.%. The catalyst exhibited a maximum activity at Co/Mo ratio 0.3. The order in which metal species were added had a great influence on the activity of the Co-Mo/gamma-Al2O3 catalyst. Higher activity was obtained when Co was added into Mo/gamma-Al2O3 as opposed to Mo added into Co/gamma-Al2O3. PMID- 12090287 TI - Using multiple criteria decision analysis for supporting decisions of solid waste management. AB - Design of solid-waste management systems requires consideration of multiple alternative solutions and evaluation criteria because the systems can have complex and conflicting impacts on different stakeholders. Multiple criteria decision analysis (MCDA) has been found to be a fruitful approach to solve this design problem. In this paper, the MCDA approach is applied to solve the landfill selection problem in Regina of Saskatchewan Canada. The systematic approach of MCDA helps decision makers select the most preferable decision and provides the basis of a decision support system. The techniques that are used in this study include: 1) Simple Weighted Addition method, 2) Weighted Product method, 3) TOPSIS, 4) cooperative game theory, and 5) ELECTRE. The results generated with these methods are compared and ranked so that the most preferable solution is identified. PMID- 12090288 TI - Estimation of atmospheric mixing heights over large areas using data from airport meteorological stations. AB - For this study, ground and sounding meteorological data at 16 airports were used for estimating the atmospheric mixing heights in Hebei Province of China, including Beijing and Tanjing areas. Three methods were compared for this purpose, which are the dry adiabatic method, the conventional Nozaki model, and a modified Nozaki model. The feasibility of using airport meteorological data to determine mixing heights in large area was verified through the use of measured mixing heights and observed SO2 vertical profiles. The study not only estimated the mixing heights over large areas but also compared different early morning sounding temperature profiles to identify feasibility of using 2:00 a.m. sounding profiles to estimate mixing height by the dry adiabatic method. The paper also described the mixing heights over different areas such as mountain, sea boundary and plain areas. The results have considerable significance for air quality prediction and environmental management. A novel approach to estimation of atmospheric mixing heights over large area is introduced, requiring only the input of readily available airport meteorological data. PMID- 12090289 TI - Monitoring C-reactive protein in beagle dogs experimentally inoculated with Ehrlichia canis. AB - The concentrations of C-reactive proteins (CRP) in the plasma of five beagle dogs experimentally inoculated with Ehrlichia canis increased markedly. The concentrations began to increase between 4 and 16 days and peaked between 15 and 42 days after inoculation of E. canis. The peak concentrations ranged from 217.8 to 788.8 microg/ml (452.6 +/- 228.1 SD). After the peak, the concentrations of CRP decreased rapidly. The PCR product of 16S rRNA of E. canis became detectable in the five dogs between 18 and 27 days after inoculation of E canis. Antibodies to E canis were detected in plasma from the dogs between 5 and 15 days after inoculation of E. canis. The timings of seroconversion and of the start of the increase in CRP were approximately similar and the high concentrations of CRP in the plasma of the dogs tended to become apparent when the PCR product of 16 S rRNA of E. canis became detectable. PMID- 12090290 TI - Antigenic relationships within the genus Salmonella as revealed by anti Salmonella enteritidis monoclonal antibodies. AB - A panel of 38 monoclonal antibodies (MAbs) that react with outer membrane proteins (OMPs) of Salmonella enteritidis was produced. On the basis of their binding pattern in ELISA, the MAbs were divided into three groups. The first group, consisting of 15 MAbs, was found to be Salmonella-specific as they did not cross-react with Escherichia coli or Pasteurella multocida. The second group of 15 MAbs cross-reacted with E. coli but not with P. multocida, reflecting the closer antigenic relationship of E. coli with Salmonella. The third group of 8 MAbs cross-reacted with both E. coli and P. multocida, indicating that the antigenic determinants identified by these MAbs are conserved in all the three genera. The antigenic relationship of the Salmonella serovars (S. enteritidis, S. gallinarum, S. typhimurium, S. dublin, S. agona, S. indiana and S. choleraesuis) was studied using OMPs prepared from them and the anti-S. enteritidis MAbs. Three MAbs appeared to be specific for S. enteritidis as they did not cross-react with any of the other Salmonella serovars. Twelve of the 38 MAbs cross-reacted with all the serovars tested. Six of these were specific to the Salmonella genus as they did not cross-react with any of the other Gram-negative bacteria tested. The reactivity pattern of the other MAbs indicated that S. gallinarum was antigenically close to S. enteritidis, followed in order by S. dublin, S. agona, S. typhimurium and S. indiana, whereas S. choleraesuis seemed to be antigenically quite distant from S. enteritidis. PMID- 12090291 TI - Clinicomorphological studies in chicks fed ochratoxin A while simultaneously developing coccidiosis. AB - The progression of coccidiosis and the resultant mortality were followed in chicks fed a OTA-contaminated diet. More complex and rapid progress of coccidiosis occurred in OTA-treated chicks than in chicks fed a OTA-free diet. The concentration of total protein in the serum was significantly decreased in the chicks in the OTA-treated group, whereas this was significantly increased in chicks infected with Eimeria tenella, irrespective of additional treatment with OTA. The serum glucose concentration was significantly increased in all the chicks exposed to OTA and/or suffering from coccidiosis, as was serum retention of uric acid in all groups, most notably in those consuming OTA. OTA induced degenerative changes in, and an increase in the weight of the kidneys, liver, heart and ventriculum; there was depletion of lymphoid tissue and a decrease in the lymphoid organs' weight and body weight. Coccidiosis induced only a slight growth depression and a slight increase in the relative weight of the kidneys and liver. The intensity of the clinical signs, the impairment of kidney function, macroscopic and histopathological changes, deviations in the weight of some organs and general depression in growth were greater when chicks infected with E. tenella were also given OTA. PMID- 12090293 TI - Demonstration of generalized infection with caprine herpesvirus 1 diagnosed in an aborted caprine fetus by PCR. PMID- 12090292 TI - Phylogenetic analysis of field isolates of feline calcivirus (FCV) in Japan by sequencing part of its capsid gene. AB - The molecular epidemiology of the infectious disease caused by feline calcivirus (FCV) in Japan was investigated by analysing the phylogenetic relationship among 21 Japanese field isolates, including the F4 strain, and 30 global isolates. Parts of the capsid gene (B-F) of the isolates were amplified by RT-PCR, and the amino acid sequences were compared with those from the global isolates. Thirty seven and 14 out of a total of 51 isolates were clustered into two distinct genogroups, I and II respectively, by UPGMA and NJ analysis. Seven of the 21 Japanese isolates (33%) fell into group I together with 30 global isolates, while the other 14 Japanese isolates (67%) belonged to group II. The bootstrap repetition analysis of groups I and II formed by the NJ method gave a value of 99.00%. The 14 latter Japanese isolates were clearly separated from the isolates in group I, and they were different from any previously known FCV, forming a new genogroup, which implies that this lineage has been confined to Japan. Comparing the amino acid sequences shared by groups I and II, the amino acid at position 377 in B region was asparagine (Asn or Asp (NH2)) in group I, while it was lysine (Lys) in all the strains in group II. Similarly, the amino acid at position 539 in the F region was alanine (Ala) or proline (Pro) in group I, while it was valine (Val) in group II; glycine (Gly) at position 557 in group I was serine (Ser) in Group II; and phenylalanine (Phe) or leucine (Leu) at position 566 in genogroup I was tyrosine (Tyr) in group II. PMID- 12090294 TI - Effect of immunization with plasmid DNA encoding for rinderpest virus matrix protein on systemic rinderpest virus infection in rabbits. AB - Plasmid vaccine pBK-CMVMPILC113 expressing the matrix (M) gene of rinderpest virus was assessed for its potential to protect rabbits against a lethal viral challenge. Rabbits immunized with plasmids expressing the M gene were not protected when challenged with lapinized rinderpest virus, despite the production of anti-M antibodies, while rabbits immunized with rinderpest tissue culture vaccine were completely protected from a lethal challenge with lapinized rinderpest virus. The plasmid vaccine also had no significant effect on the lymphopenia in challenged rabbits. The results indicate that rinderpest M protein does not have a protective role in rinderpest infection. PMID- 12090296 TI - An experimental study on the perineurial window. AB - Neurological symptoms of herniated nerve fibers resulting from limited perineurial injury from sharp materials such as needles have become a recent topic in clinical practice. However, the mechanism of this disorder, which is known as a perineurial window, has not been clarified. To investigate the mechanism of nerve damage in the perineurial window, we designed small (1-mm length) and large (5-mm length) perineurial windows using tibial nerves of Wistar rats. In the 1-mm group, a marked hernia of the endoneurial contents developed soon and decreased in size with time, but protrusion of nerve fibers was still observed after 12 weeks. Nerve fibers in both the herniated portion and under the edge of the window were damaged. Even after 12 weeks, regeneration of the nerve fibers and the perineurium was incomplete. In contrast, in the 5-mm group, the initial endoneurial edema was remarkable, but herniated nerve fibers were not seen after 12 weeks. Neurological impairment in the 5-mm group was marked in the early stage but rapidly recovered. The repair of the perineurium and nerve fibers in the 1-mm group was slower than in the 5-mm group. Persistent neurological symptoms in the perineurial window appeared to be more closely associated with entrapment of nerve fibers at the window edge rather than with disruption of endoneurial homeostasis. PMID- 12090295 TI - Bovine immunodeficiency virus expression in vitro is reduced in the presence of beta-chemokines, MIP-1alpha, MIP-1beta and RANTES. AB - The inhibition of HIV expression in vitro by a cocktail of the beta-chemokines MIP-1alpha, MIP-1beta and RANTES provided the initial evidence that HIV utilizes chemokine receptors as co-receptors for infection of cells. Bovine immunodeficiency virus (BIV), a lentivirus, infects a wide variety of leukocyte populations, but the cellular receptor(s) utilized by this virus for infection of cells is not known. The purpose of this study was to determine whether MIP 1alpha, MIP-1beta and RANTES affect BIV expression in vitro, as a prelude to identifying the cellular receptors utilized by this virus. Fetal bovine lung (FBL) cells were pretreated with serial dilutions of a cocktail of the chemokines, and then the cells were infected with BIV. Virus expression in these cells was determined by counting the syncytia that had developed in the cultures by five days after infection. A significant decrease in syncytium formation, corresponding to increasing concentrations of the chemokines, was the result. Reacting the chemokines with chemokine-specific neutralizing antibodies prior to treatment of the cells neutralized the effect of the chemokines on virus replication in a dose-dependent manner, restoring viral expression to a level similar to that of untreated cells. The presence of a CCR5 homologue on the surface of FBL cells was confirmed using an anti-CCR5 monoclonal antibody and FACS analysis. Collectively, these data provide preliminary evidence that BIV may utilize the CCR5 receptor for infection of cells in vitro, but additional studies are necessary to confirm this. PMID- 12090297 TI - Clinical versus quantitative vibration assessment: improving clinical performance. AB - In 3 large cohorts (total of 787 patients), clinical vibration impairment (CVI) of the great toe using a tuning fork was compared with quantitative vibration threshold (QVT). Using a stepwise multivariate analysis, we assessed demographic and anthropomorphic patient characteristics associated with the difference between CVI and QVT for the various cohorts and the chosen QVT ranges of percentile abnormality. We also compared CVI or QVT abnormality with a composite score of nerve conduction abnormality to confirm that QVT is a valid measure of severity of neuropathy. Highly significant associations between CVI and QVT were found in all 3 cohorts studied, regardless of the chosen QVT percentile level of abnormality. However, in the 2 cohorts evaluated by many different physicians, CVI overestimated QVT much more often than underestimated it. The discordance between CVI and QVT in all QVT abnormality percentile levels was associated with age, height and body surface area (BSA) in 1 cohort, with age and BSA in another cohort, and with age in the third cohort. In the third cohort, the correlation between QVT and the composite score of nerve conduction abnormality was significantly higher than the correlation between CVI and the composite score. Using a tuning fork, neuromuscular physicians overestimate vibration sensation loss more often than when QVT testing is done, which employs quantitative stimuli, a broad range of stimulus magnitudes, null stimuli, validated algorithms of testing and validated reference values. To improve assessment of vibration sensation, physicians should take into account age, height and weight (or body surface area) when judging vibration abnormalities. Applying some useful approaches to quantitative sensory testing might improve the accuracy of clinical sensory testing. PMID- 12090298 TI - Local administration of vasoactive intestinal peptide after nerve transection accelerates early myelination and growth of regenerating axons. AB - Our goal was to determine whether local injections of vasoactive intestinal peptide (VIP) promote early stages of regeneration after nerve transection. Sciatic nerves were transected bilaterally in 2 groups of 10 adult mice. In the first group, 15 microg (20 microL) of VIP were injected twice daily into the gap between transected ends of the right sciatic nerve for 7 days (4 mice) or 14 days (6 mice). The same number of mice in the second group received placebo injections (20 microL of 0.9% sterile saline) in the same site, twice daily, for the same periods. After 7 days, axon sizes, relationships with Schwann cells and degree of myelination were compared in electron micrographs of transversely sectioned distal ends of proximal stumps. Fourteen days after transection, light and electron microscopy were used to compare and measure axons and myelin sheaths in the transection gap, 2-mm distal to the ends of proximal stumps. Distal ends of VIP-treated proximal stumps contained larger axons 7 days after transection. More axons were in 1:1 relationships with Schwann cells and some of them were surrounded by thin myelin sheaths. In placebo-treated proximal stumps, axons were smaller, few were in 1:1 relationships with Schwann cells and no myelin sheaths were observed. In VIP-treated transection gaps, measurements 14 days after transection showed that larger axons were more numerous and their myelin sheaths were thicker. Our results suggest that in this nerve transection model, local administration of VIP promotes and accelerates early myelination and growth of regenerating axons. PMID- 12090299 TI - LR white post-embedding colloidal gold method to immunostain MBP, P0, NF and S100 in glutaraldehyde fixed peripheral nerve tissue. AB - A variety of immunocytochemical techniques are now widely used for the electron and light microscopic examination of biological samples. They are employed routinely for investigating the role of certain proteins in nervous tissue. Immunoelectron microscopic studies require the tissue to be fixed and embedded in a solid support, which may disrupt cellular structures and destroy crucial antigens. A technique of post-embedding with LR white resin has been developed, and it has been shown that certain antigens tolerate fixation with glutaraldehyde. In this study, we optimized a previous post-embedding method using low-water-miscible low-temperature embedding resin (LR white) to immunostain MBP, P0, NF and S100 proteins in peripheral nerves fixed with a relatively high concentration of glutaraldehyde found to be compatible with the morphology of normally compacted nerve fibers from humans and adult animals. The main difference in the procedures described here from previous ones is the elimination of vibratome sectioning, rendering this immunostaining technique more accessible to neuropathological laboratories using standard equipment for the ultrastructural study of peripheral nerves. It may prove of value for localization and quantification of these proteins in normal and pathological conditions. PMID- 12090300 TI - Mutation analysis of 12 candidate genes for distal hereditary motor neuropathy type II (distal HMN II) linked to 12q24.3. AB - Distal hereditary motor neuropathies (distal HMNs) are characterized by degeneration of anterior horn cells of the spinal cord resulting in muscle weakness and atrophy. Distal HMN type II is genetically linked to chromosome 12q24.3 and located within a 13 cM region flanked by D12S86 and D12S340. We previously excluded 5 positional and functional candidate genes for distal HMN II. Here, we report the exclusion of 12 additional candidate genes localized within the distal HMN II region; the genes include musashi (Drosophila) homolog 1 (MSI1), protein inhibitor of neuronal nitric oxide synthase (PIN), peripherin (PRPH), tubulin alpha ubiquitous (K-ALPHA-1), tubulin alpha 3 (TUBA3), tubulin alpha 6 (TUBA6), splicing factor arginine/serine-rich 9 (SFRS9), U5 snRNP 100 kd (U5- 100K), putative chemokine receptor, GTP-binding protein (HM74), MondoA, cut (Drosophila)-like homeobox 2 (CUX2) and ADP-ribosylation factor 3 (ARF3). PMID- 12090301 TI - Assessment of the "common" 4.8-kb mitochondrial DNA deletion and identification of several closely related deletions in the dorsal root ganglion of aging and streptozotocin rats. AB - The identification of several mitochondrial DNA (mtDNA) deletions and the accumulation of the "common" 4.8-kb mitochondrial DNA deletion (mtDNA(4834)) with aging and experimental streptozotocin-induced diabetes (STZ) were studied in the rat dorsal root ganglion (DRG). Twenty-one mtDNA deletions, including mtDNA(4834), were identified in rat L4-L6 DRG mtDNA of 15-month-old Spraque Dawley rats with 13 months of STZ and age-matched controls. These deletions were flanked by breakpoints that ranged from 16-bp direct repeats to no direct repeats. The sciatic nerve contained undetectable levels of mtDNA deletions. Levels of mtDNA(4834) in rat DRG mtDNA significantly accumulated with age at a rate much higher than those reported in the brain, yet were not statistically different in STZ. Southern blot analysis demonstrated no significant accumulation of the total amount of mtDNA deletions in STZ over age-matched controls. The accumulation of mtDNA(4834) has not been studied in rat peripheral nerve tissue. Our identification of several mtDNA deletions with and without direct repeats at their breakpoint support the hypothesis that deletions can occur by both the slip replication model and random recombination. Although there is a significant increase in accumulation of mtDNA(4834) associated with aging, the lack of significant accumulations of mtDNA deletions in STZ over age-matched controls indicates that this type of mtDNA damage is likely not a major alteration in STZ, although the changes could be confined to a small population of neurons that undergo apoptosis between 8 and 15 months. PMID- 12090302 TI - Crisis intervention program: an alternative to inpatient psychiatric treatment for children. AB - This study evaluated the impact of a Crisis Intervention program as an alternative to use of psychiatric treatment beds for young children. A multidisciplinary community-based intervention was utilized, including family therapy, psychiatric intervention, and school consultations. The impact of the service was evaluated in relation to the use of psychiatric treatment beds by the population of children eligible for Medicaid or uninsured. In comparison to an historical control group, the program resulted in a 23% reduction in the use of psychiatric treatment beds. A cost-minimization analysis indicated that in addition to the program reducing the use of psychiatric treatment beds, the cost of treatment was also slightly reduced. PMID- 12090303 TI - Community integration of persons with psychiatric disabilities in supportive independent housing: a conceptual model and methodological considerations. AB - Despite the consensus regarding community integration as a major goal of mental health policy and the emergence of supportive independent housing as a critical component of community mental health services, mental health services research has not examined the extent to which housing and service characteristics are associated with community integration of persons with psychiatric disabilities in supportive independent housing. The main goal of this paper is to propose a conceptual model of factors influencing community integration that takes into account the differential configuration of housing setting and support structure in supportive independent housing. The conceptual model encompasses a multidimensional conceptualization of community integration and considers an array of housing and service characteristics that are potentially relevant determinants of community integration. On the basis of the proposed model, this paper outlines the methodological considerations for future research with regard to measurement, research designs, and statistical models. PMID- 12090304 TI - Adapting to psychiatric disability and needs for home- and community-based care. AB - The objective of the study was to describe adaptation strategies and use of formal and informal support by individuals with psychiatric disabilities, to delineate remaining needs, and to determine how home- and community-based services might address those needs. Using in-depth interviews and structured questionnaires, we examined functional status, adaptation, and needs for home- and community-based care among 33 severely mentally ill members of a large health maintenance organization. Despite success in community living, participants had significant functional deficits (physical and emotional), relied heavily on only one or two key informal caregivers, and often needed significant support from mental health professionals. Limited numbers of caregivers and social isolation placed participants at risk of negative outcomes if informal support resources were to be lost. Home- and community-based care interventions that attempt to increase informal support networks and provide instrumental help (cooking, cleaning, transport) on short notice during flare-ups could augment existing (but limited) informal caregiving, help severely mentally ill individuals remain independent, and reduce the likelihood that loss of an informal caregiver would result in unwanted outcomes. PMID- 12090305 TI - Impact of services, met needs, and service empowerment on consumer outcomes. AB - The current study examined service characteristics, needs, and outcomes of consumers with severe mental disabilities served in a public mental health setting. The study utilized a longitudinal design with three yearly waves of measurement. Data were obtained from consumer interviews and case manager questionnaires. A model specifying relationships between service characteristics, needs, and outcomes was tested using structural equation modeling. This model incorporated service-related variables pertaining to service amounts, service empowerment, and needs in order to gain a better understanding of factors that mediate service effects and account for mental health outcomes. The model focused on consumers' perspectives, in order to highlight the importance of consumers' opinions about their mental health services and needs. The results suggested that consumers' perceptions that their needs are met were related to better symptomatology and quality of life outcomes. In addition, consumers' perceptions that they had some say in service-related decisions had an indirect effect on mental health outcomes by increasing the likelihood that needs were met according to consumers' perspectives. Results were mixed with regard to the effects of these variables on level of functioning outcomes. Service amount was unrelated to mental health outcomes. Furthermore, service amount was unrelated to consumers' perceptions of needs, suggesting that consumers' views regarding met and unmet needs may not be included sufficiently in decision-making surrounding the provision of services. PMID- 12090306 TI - Effect of geographic migration on SMI prevalence estimates. AB - The prevalence of serious mental illness (SMI) varies by the socioeconomic characteristics of communities. This variation is presumed to be due to the differential incidence of disorders caused by adverse social factors (social causation) and differential geographic migration (social selection and drift). The objective of this study was to measure the geographic migration patterns of adults treated for SMI. A sample of 11,725 adults with three or more psychiatric hospital admissions between July 1978 and November 1992 was drawn from inpatient records. At least one third migrated to different counties between first and last admission. Migration rates were higher for Whites than for African Americans and higher for unmarried than for married patients. There were no significant differences in migration rates by gender or mental disorder. Patient migration did not parallel shifts in the general population. Patient migration was generally toward medium-size, low-income urban counties with relatively declining general populations, and movement was away from both the most rural and the largest urban counties. Public needs assessments and resource allocation policies may understate the need in communities with net SMI out-migration and overstate endemic need where there is net in-migration. In the long term, these same policies may induce migration through resource allocation decisions. PMID- 12090307 TI - Diagnosing depression and prescribing antidepressants by primary care physicians: the impact of practice style variations. AB - This study examined variations in the diagnosis of depressive disorders and prescription of antidepressant medications in primary care and the contribution of the physicians' practice styles to these variations. The analyses were based on visits to a representative sample of the U.S. office-based primary care physicians from the 1997 and 1998 National Ambulatory Medical Care Survey. In this sample, the propensities to give a diagnosis of depressive disorder and to prescribe antidepressants were operationalized as propensity variables. The association of these variables, obtained from a randomly selected subsample of visits to each physician, with the diagnoses and treatments of the other patients seen by the same physicians was examined. The results revealed considerable variations across practices in the percentages of patients who received diagnoses of depressive disorders (0-25%) and prescriptions for antidepressant medications (0-38%). Furthermore, the physicians' propensities to diagnose depressive disorders or to prescribe antidepressants were significantly associated with the diagnosis and treatment of individual patients. The large variations in diagnosis and treatment of depressive disorders and the significant impact of practice style variables on these variations highlight the need for implementation of uniform practice guidelines for diagnosis and treatment of depressive disorders in primary care settings. PMID- 12090309 TI - Contextual factors affecting job satisfaction and organizational commitment in community mental health centers undergoing system changes in the financing of care. AB - This study examines the relationship between contextual factors and job satisfaction and organizational commitment among a sample of 148 administrators and staff in 17 community mental health centers undergoing the transition from fee-for-service (FFS) reimbursement to capitation of Medicaid-funded mental health services in Colorado. Hierarchical linear modeling was used to assess both organizational level factors as well as factors at the individual level of analysis in relation to job satisfaction and organizational commitment. Results indicated significant associations between type of financing for Medicaid-funded services and organizational commitment. Direct, nonprofit capitation was positively linked to organizational commitment, relative to traditional FFS reimbursement. Aggregate perceptions about the organization, including its culture and climate, and the formalization of policies and procedures were strongly linked to job attitudes, over and above individual perceptions about the organization. Individual perceptions of the organization were also related to job attitudes as was respondent level within the hierarchy of the organization. These findings are discussed in terms of their implications for research and intervention in mental health service settings. PMID- 12090308 TI - Dose response in child and adolescent mental health services. AB - This study examines the dose-response relationship, the correlation between the amount of mental health treatment a child receives (dose) and the outcome (response) in a community setting. Participants were 125 children treated in the Stark County Child and Adolescent Mental Health System. Study methods include multiple outcomes, multiple-dose definitions, longitudinal hierarchical analysis of repeated measures, and instrumental variable estimation to control for possible confounding between outcome and treatment dose. Results show no statistically significant dose response. The results do not support the existence of a dose response for children and adolescents consistent enough to guide clinicians, administrators, or policymakers. PMID- 12090310 TI - Treatment programs for youth with emotional and behavioral disorders: an outcome study of two alternate approaches. AB - Youth with severe emotional and behavioral disorders (EBD) were randomly assigned for 3 months of intensive treatment to a 5-day residential program (5DR Program) or a community-based alternative, family preservation program (FP Program). Programs differed not only in method of service delivery (residential unit vs. home-based), but also in treatment philosophy (solution focused brief therapy vs. cognitive behavioral). Results confirmed high rates of comorbidity in this population for externalizing and internalizing disorders. A significant Treatment x Program interaction was evident for internalizing disorders. At 1-year follow up, significantly higher percentages of youth from the FP Program revealed a reduction of clinical symptoms for ADHD, as well as, general anxiety and depression, whereas significant proportion of youth from the 5DR Program demonstrated clinical deterioration and increased symptoms of anxiety and depression. Results have implications for future treatment of youth with EBD and suggest that greater emphasis be placed on research linking treatment to specific symptom clusters, especially highly comorbid clusters in this hard to serve population. PMID- 12090311 TI - Methodological challenges in assessing children's mental health services utilization. AB - This paper describes the reliability and validity of the service assessment for children and adolescents (SACA) for use among Spanish-speaking respondents. The test-retest reliability of the instrument was assessed in a randomly selected clinical sample of 146 Puerto Rican children and adolescents aged 4-17. Both parents and children were administered the SACA twice by independent interviewers over an average 12-day follow-up period. The accuracy of parental and youth self reports was assessed by comparing these reports to information obtained from medical records. The results showed that parents and children (aged 11-17) were able to report with fair to moderate reliability any last year use of mental health services, any outpatient mental health services, and school services. Residential and hospitalization services were reported by both informants with substantial test-retest reliability. Slight or no test-retest reliability was obtained for parent and child on the use of the specific type of mental health professionals, as well as parental reports of several treatment modalities. Substantial sensitivity of the SACA was obtained when comparing medical records to parental and child reports to lifetime use of any service and outpatient mental health service. Moderate sensitivity was obtained for last year use of mental health services for both parent and child informants. PMID- 12090312 TI - Case report: testicular secondary localization of a multiple myeloma. AB - A 61-year-old man who had 40 months history of recurrent multiple myeloma presented a right testicular mass, which was diagnosed incidentally by scrotal sonography examination. Alpha-feto-protein and beta-human-chorionic-gonadotropin were normal. A right radical orchiectomy was performed. The histological examination showed an intermediate grade plasmacytoma. PMID- 12090313 TI - Leydig cell tumour--a rare testicular tumour. AB - Although Leydig cell tumour is a rare tumour which constitutes only 1-3% of all testicular tumours, still it is in the focus of interest because of the difficulties in determining its exact nature and subsequently the type of treatment and follow-up. We report a case of Leydig cell tumour with a review of the related literature. PMID- 12090314 TI - Clinical experiences with different one-staged surgical methods for primary hypospadias cases. AB - The success of various types of operations for primary hypospadias was assessed taking the location of meatus and the degree of chordee as the basis. Totally 258 patients with primary hypospadias were operated: 94 by meatal and urethral advencement, 110 by perimeatal flap and tube repair and 54 by preputial flap. The rates of success in cases with meatal advencement, perimeatal flap and preputial flap were found as 88%, 81% and 66% respectively. The success of treatment in hypospadias cases is closely related to the age of the patient, the experience of the surgeon and the choice of an appropriate method. However, the presence of chordee and the proximally located meatus are among the leading factors that influence the rate of success. PMID- 12090315 TI - Distal hypospadias repair: comparison of two different techniques. AB - The cosmetic results and the morbidity of Mathieu and Barcat techniques are presented and compared in a series of 52 children aged between five months to 19 years during January 1991 to May 2001. All patients treated with the Mathieu procedure had negligible glandular hypospadias at some level. The neomeatus was slit-like or elliptic in majority of the cases. The horizontal bucket handle deformity was eliminated which remains as a meatal problem of the original Mathieu procedure. Four patients had fistula, which was closed by surgery whereas another three had mild stricture formation. Cosmetic and functional results were excellent with the Barcat balanic groove technique. There was no fistula formation, only two had meatal stricture which responded to dilatations. The Barcat technique allows anatomically superior glans reconstruction resulting in a vertical and slit like neomeatus in distal hypospadias repair with a low complication rate. PMID- 12090316 TI - Microvascular replantation of a self amputated penis. AB - In this case report, microvascular replantation of a self amputated penis is reported. The reasons of being unsuccessful were discussed and the literature was reviewed. PMID- 12090317 TI - Increased incidence of neoplasia in chronic renal failure (20-year experience). AB - Patients with chronic renal failure (CRF) have a high incidence of malignant tumours. Uremia thus induces a remarkable suppression of immune status. In this study, we report that within the last 20 years, 188 (6.7%) various organ tumours were found in 2817 CRF patients that were hospitalised and treated. 69 (36.7%) of 188 patients with various organ tumours were on hemodialysis and the rest (63.3%) were CRF without hemodialysis. 49 (71%) of the 69 patients with hemodialysis were diagnosed with tumours in the first year of the hemodialysis therapy. In 110 (84%) of 119 CRF patients tumours were detected in less than 10 years after diagnosis of CRF. Localisation of the tumours were: 39 (19%) urologic malignancy, 30 (16%) parathyroid adenoma, 28 (15%) skin malignancy, 19 (10%) gynaecologic malignancy, 18 (9.5%) gastrointestinal tract (GIT) malignancy, 17 (9%) lung malignancy, 17 (9%) larynx malignancy, 13 (7%) thyroid malignancy, 5 (2.6%) multiple myeloma and 5 (2.6%) malignant lymphoma. No patients in the series had received a transplant kidney or were therapeutically immunosuppressed for other reasons and obstructive uropathy. CRF patients have a several times greater risk of developing malignant tumour than the general population. PMID- 12090318 TI - Experimental study on effects of deferoxamine mesilate in ameliorating cisplatin induced nephrotoxicity. AB - PURPOSE: Cisplatin (CCDP), an indispensable agent of several chemotherapy protocols, has serious dose limiting side effects, including nephrotoxicity. In this experimental study, we used deferoxamine mesilate (DFO), an iron chelating agent, to ameliorate cisplatin-induced nephrotoxicity. MATERIALS AND METHODS: Sixty adult male bulb-c mice were divided in 6 equal groups. Group 1 received distilled water, group 2 received 100 mg/kg DFO, group 3 received 0.9 mg/kg CCDP, group 4 received 100 mg/kg DFO one hour before 0.9 mg/kg CCDP, group 5 received 1.8 mg/kg CCDP, and group 6 received 200 mg/kg DFO one hour before 1.8 mg/kg CCDP transperitoneally for 10 days. The next day, blood and urine samples were obtained, and all the animals were sacrificed, the kidneys and testes were removed, and histopathologic and biochemical analyses were performed. RESULTS: Low-dose and high-dose CCDP treated mice had significantly more extensive proximal tubular degeneration (p < 0.001) when compared to control animals. Moreover, these changes were significantly less extensive in the mice taking DFO than mice taking CCDP. DFO showed no effect on cisplatin induced testicular histopathology. The cisplatin administration significantly increased the serum urea and plasma creatinin concentrations, and DFO administration prior to CCDP significantly decreased serum urea and plasma creatinin concentrations. CONCLUSION: Our findings suggest that DFO administration may be safe and useful for ameliorating cisplatin-induced nephrotoxicity. PMID- 12090319 TI - Is there a need for pelvic CT scan in cases of renal cell carcinoma? AB - OBJECTIVES: To determine the necessity of pelvic computed tomography (CT) in patients of renal cell carcinoma (RCC). MATERIALS AND METHODS: We reviewed the records of 400 patients of RCC, who underwent treatment at our institution between January 1988 and February 2001. These patients were evaluated pre operatively with ultrasonograms (USG) and contrast enhanced CT scan of the abdomen and pelvis. USG or CT scans of these cases were reviewed for presence of pathology in the pelvis, which were classified into 3 categories viz; benign and likely to be insignificant, benign and likely to be significant; and malignant. RESULTS: Of the 400 cases, 114 were stage I, 68 were stage II, 99 were stage III and 119 were stage IV. In all patients, tumour was identified in the kidney on preoperative CT scan. Fourteen patients (3.5%) had an abnormality on pelvic CT. Five (1.25%) had category 1, three (0.75%) had category 2 and six (1.5%) had category 3 abnormality on pelvic CT. However, all these abnormalities in pelvis were detected prior to CT by other investigations (USG or plain x-ray). Of the six cases with malignant findings, two had superficial bladder cancer, one had RCC in a pelvic kidney and three had bone metastases in the pelvis. CONCLUSIONS: Pelvic CT does not offer additional information in the vast majority of cases with RCC and should be performed selectively. Thus the cost of diagnostic imaging in RCC can be reduced. PMID- 12090320 TI - Acute renal failure in leptospirosis in the black-sea region in Turkey. AB - Leptospirosis is an infectious disease caused by pathogenic leptospires and is characterized by a broad spectrum of clinical manifestations, varying from inappearent infection to fulminant, fetal disease. Eighty-five to 90% of leptospirosis infections are self-limiting. However, 5-10% of infection by L. interrogans can cause renal tubular damage, microvascular injury, acute renal failure (ARF), and interstitial nephritis. We studied 36 patients with leptospirosis. Twenty-seven (65%) cases of 36 patients had ARF. Fourteen (51%) had nonoliguric ARF. In thirteen (48%) oliguria appeared on the third or fourth days of hospitalization. Serum BUN, creatinine, serum bilirubine, ALT, AST, potassium and thrombocytopenia levels were higher in oliguric than nonoliguric patients (p < 0.05). However, serum sodium, CPK levels were not different between oliguric and nonoliguric groups (p > 0.05). Thirteen patients (48%) needed in renal replacement therapy (RRT). 8 of them were treated by hemodialysis (HD) alone and 5 patients by HD in combination with hemoperfusion. Twenty-five patients (92%) recovered completely after 3-5 weeks. Two patients (7.4%) who had severe hepatorenal and hemorrhagic syndromes, died. We concluded that till now leptospirosis is actual problem for nephrologist in the developing countries because of very high percentage of renal disease, with good prognosis in patients without multiorgan failure and early treatment. PMID- 12090321 TI - Staff-assisted home hemodialysis in debilitated or terminally ill patients. AB - End stage renal disease (ESRD) patients who are diagnosed to have a terminal illness or severe debility have limited options for their continued care. This results in a frequent decision to withdraw dialysis support. Due to their tenuous condition, continued transportation to the dialysis facility further aggravates the emotional, financial and physical burden to the patient and family. We would like to present our data on 28 patients with severe debilitating and terminal illnesses. The mean age was 69 years with a (+/-) 11.8 SD and range of 44-87 years. Nine of them were males and 19 females. All of these were considered terminally ill as most of these patients had multi-organ failure. Ten had stroke, 16 had cardiac failure, 2 had severe vascular insufficiency, one resulting in bilateral leg amputation, 5 had debilitating pulmonary disease needing oxygen therapy and 8 had cancer. These patients were dialyzed at their home by a registered nurse (RN) according to a dialysis prescription provided by an attending nephrologist. Twenty-three patients died at home, one transferred to acute care facility and 3 to hospice care after a mean staff-assisted home hemodialysis (SAHD) duration of 14.1+/-2.9 weeks. ESRD patients with severe disability can continue dialysis in a more convenient and comfortable setting at home, and yet be relatively cost-effective. PMID- 12090322 TI - Acute rejection in the elderly recipient: influence of age in the outcome of kidney transplantation. AB - Since the immune response in older recipients is weaker they should be less likely to reject a transplanted organ and should need less aggressive immunosuppressive treatment. Our aim was to record the incidence and severity of episodes of acute rejection (AR), estimate the influence of these events on graft survival of elderly recipients (> or = 60) and to compare these with that in younger ones. We performed 363 kidney transplants between 1/94 and 12/98, and recorded clinical and immunological data, incidence-severity of AR and cause of graft loss. Patients were divided into two groups, according to the age at transplantation: A (<60, n = 281/77.4%) and B (> 60, n = 82/22.6%). The percentage of aging recipients and mean age of donors and recipients increased throughout the period. Although the incidence of ATN was higher in the older group (29% vs.19%, p < 0.0001) the number of graft biopsies was equal in both groups. The incidence of AR was similar, 33.4% vs. 26.8%, pNS. The number of AR episodes per patient was 0.44 and 0.41 respectively. The severity of AR was: Banff grade I: A (40.3%)/B (45.7%) pNS; grade II: A (44.1%)/B (48.57) pNS; grade III: A (15.5%)/B (5.7%) pNS. Younger recipients presented a higher level of panel reactive antibodies (PRA) (4.3% vs. 2.07%, p = 0.01). One-yearpatient survival was 96%/91% (p < 0.05) and graft survival was 81%/78% (pNS) respectively. The age of recipient does not seem to have influenced the incidence-severity of AR or the graft survival. Thus immunosuppression should be individualized for each patient and should not depend on the age at transplantation. PMID- 12090323 TI - Oliguric acute renal failure in mycosis fungoides with lymphomatous infiltrates in the kidneys. AB - OBJECTIVE: To present the clinical picture of acute renal failure in patients with mycosis fungoides (MF) and renal lymphomatous infiltrates. To analyze the pathogenesis of renal failure. METHODS: Correlation of clinical picture, urinary findings, imaging reports and autopsy findings in two patients with long-standing MF who died with renal failure. CASE SUMMARIES: Both subjects had sustained oliguria in the last 2 weeks. One patient had persistent hypotension, normal urinalysis, normal renal sonogram, and scarce interstitial lymphomatous infiltrates with preservation of renal parenchymal architecture. He was thought to have ischemic acute renal failure not directly linked to the lymphomatous infiltrates. The second patient developed hypertension one month prior to death, and had moderate proteinuria, hematuria, pyuria, grossly enlarged kidneys with hypoechoic masses, and extensive replacement of the renal parenchyma by lymphomatous infiltrates. This picture is typical of renal failure secondary to lymphomatous replacement of the kidneys. CONCLUSIONS: The development of oliguric renal failure in MF with renal lymphomatous infiltrates may have varying clinical and imaging manifestations and pathogeneses. Potentially reversible pathogenic mechanisms should be systematically investigated, particularly if the overall clinical picture is not characteristic of renal failure secondary to lymphomatous replacement of the parenchyma. PMID- 12090324 TI - Characteristics and outcome of membranous nephropathy in older patients. AB - Many patients with idiopathic membranous nephropathy are elderly, but little is known about the natural or treated history of these patients. We have studied a cohort of 155 patients with membranous nephropathy who were recruited and followed-up over a 20 year period. We have compared the clinical features and outcome of the older (>60 years) and younger age groups. There was a higher incidence of an identifiable cause for the nephropathy in older patients. At presentation with idiopathic disease, older patients were more often hypertensive and had worse renal impairment than the younger cohort, but had a similar levels of proteinuria, hypoalbuminemia and hematuria. Thrombotic complications and minor rheumatological complaints were more common in the older patients. Prognosis for life and renal survival was worse in the older onset patients. Treatment was well tolerated in selected older patients and was associated with a better outcome in those selected for treatment. PMID- 12090326 TI - Case report: percutaneous nephrolithotomy for an ectopic kidney with stone disease. PMID- 12090325 TI - Efficacy of immunosuppressive therapy in IgA nephropathy presenting with isolated hematuria. AB - The role of immunosuppressive therapy in the management of IgA nephropathy (IgAN) remains controversial. No consensus has yet emerged on the specific treatment of IgAN and this is mostly related to the lack of complete understanding of the multifactorial pathogenesis of the disease. Choice of appropriate therapeutic agents is further limited by the difficulty in identifying patients who would most likely benefit from therapy. Immunosuppressive therapy has not been recommended in patients with isolated hematuria and well preserved renal function because of their generally favourable prognosis and there are no clinical trials in this area. Considering that mild IgAN may be an early stage of the disease and can be reversed by immunosuppressive agents we have used prednisolone and azathioprine in patients with isolated hematuria in a prospective, randomized, controlled study since 1988. In this prospective study we have evaluated the effect of prednisolone with azathioprine on the clinical course of IgAN and its impact on histologic parameters and prevention of progression in patients with isolated hematuria. We studied 43 biopsy-proven IgAN patients (29 males and 14 females, aged between 13-63 years, mean age 28+/-6) with isolated hematuria and well-preserved renal function (Ccr 89.2+/-10.2 ml/min). The patients were assigned to two groups: 21 patients received prednisolone (40 mg/day) and azathioprine (100 mg/day) orally for four months (group A) and 22 patients received no specific treatment for IgAN and served as a control group (group B). In Group A prednisolone was reduced to 20 mg/day at the end of the second month, then slowly tapered over a two-month period and stopped. The median duration of follow-up was 60 months (range 12-120 months). At the end of the therapy hematuria disappeared in 17 patients. In three patients who did not respond to therapy, microscopic hematuria persisted. Of the 22 patients of group B, three had episodes of gross hematuria and proteinuria >500 mg/day. No significant changes in biochemical profile were observed in either group. Thirteen patients (eight from the treated, five from the untreated group) underwent a repeat biopsy after 12+/-6 months (range 10-25). An improvement of histopathological features was noted in Group A, while deterioration was noted in Group B. We conclude that early treatment with prednisolone and azathioprine appears to be beneficial in preventing the progression of immunologic renal injury and in improving histopathological features in IgAN patients with isolated hematuria. PMID- 12090327 TI - Significance of age in the survival of diabetic patients after kidney transplantation. AB - BACKGROUND: In recent years acceptance of diabetic patients for renal replacement therapy has increased. Renal transplantation for Type I diabetic patients is widely accepted but the appropriate treatment for Type II diabetic patients is still a matter of dispute. Our study was done to determine whether the age of Type II diabetic patients constituted an additional risk factor. METHODS: We analyzed the outcome of renal transplantation in 56 diabetic patients, 31 Type I and 25 Type II diabetics (we excluded any who had combined kidney-pancreas transplants). We compared them with 51 non-diabetic patients who were transplanted because of end-stage renal failure due to nephrosclerosis and age matched to type II diabetic patients. We assessed the one- and three-year patient and graft survival, the quality of renal function, the main complications and causes of mortality. RESULTS: The overall one- and three-year patient survival was 69% and 60% in Type II patients; 73% and 69% in Type I diabetes patients and 88% and 80% in patients with nephrosclerosis. The overall one- and three-year actuarial graft survival was 50% and 38% in patients with Type II disease and 58% and 50% in Type I diabetes, and 76% and 64% in nephrosclerosis. The main cause of graft loss in all groups was death (with functioning kidney) due to infections and cardiovascular complications. CONCLUSIONS: Diabetes itself is the most important variable in patients who have poor results after kidney transplantation. Increasing age increases slightly the risk for poor graft and patient survival. Both groups of diabetic patients have poorer results than controls but in this comparison age was an independent factor. PMID- 12090328 TI - Female nocturia. AB - Many elderly experience nocturia to such a degree that it influences their quality of life. Different studies use equivocal definitions when trying to analyze data on nocturia. The literature on etiology of female nocturia is sparse, whereas male nocturia has been studied rather extensively because of the association with lower urinary tract symptoms (LUTS). At present the few studies on female nocturia questions the fact that male and female nocturia have identical etiologies. This review will go through the present reports on prevalence and etiology of nocturia among women with reference to the male situation. PMID- 12090329 TI - Past, present and future of erythropoietin use in the elderly. AB - More than a decade has passed since the first patient with end-stage renal failure was treated with erythropoietin (EPO) and more than 85% of patients now receive this therapy. In the year 2002 more than 60% of dialysis patients will be elderly, and the treatment of anemia will be more complex due to the aditional causes: folate, iron and vitamin deficiency in this population. Correction of anemia with EPO brings about partial regression of left ventricular hypertrophy and some data suggest that such treatment reduces cardiovascular mortality in patients without advance cardiac disease. Normalization of hematocrit with EPO increases oxygen supply to the brain tissue with improvement in brain function. The improvement in the ability to recognize, discriminate and hold stimuli in memory for difficult tasks is particularly important for elderly people. No differences have been noted in the incidence of clotting of vascular access in patients treated with EPO compared with hemodialysis patients not so treated. Also no one has demostrated that treatment with EPO accelerates renal decline in patients with progressive renal insufficiency. In elderly people with anemia secondary to advanced renal failure, EPO therapy improves physical, cognitive and sexual function, and health related quality of life. PMID- 12090330 TI - Retrocaval ureter and associated abnormalities. AB - OBJECTIVE: To investigate the association of retrocaval ureter with other congenital abnormalities. METHODS AND PATIENTS: Two new cases of retrocaval ureter are discussed and a detailed electronic search of the literature was mainly focused on the concomitant abnormalities. The first of our patients had also a glandular hypospadias and a supernumerary lumbar vertebra, while the second one had syndactylia (fixed toes) in both feet. Results. The review of the literature revealed that 21% of the cases of retrocaval ureter present with concomitant abnormalities mainly from the cardiovascular system and the genitourinary tract. CONCLUSION: Diagnosis of retrocaval ureter should increase the awareness of the responsible physician on the possibility of concomitant malformations where treatment could prevent future symptoms. PMID- 12090332 TI - Comparison of lipid and lipoprotein profiles in long-term chronic ambulatory peritoneal dialysis (CAPD) in elderly patients with chronic renal failure (CRF). PMID- 12090331 TI - New trends on health related quality of life assessment in end-stage renal disease patients. AB - Health related quality of life (HRQOL) is a useful measure in the analysis of the output of medical interventions in groups of chronic patients like end-stage renal disease (ESRD) patients. From the 1980s to the present day, the interest in evaluating the HRQOL of ESRD patients has been continually growing. A review of published studies shows the existence of important differences in the applied conceptual model of HRQOL, in the assessment instruments used, in the methods of data collection recording, and in the results and conclusions. In this paper we tried to answer some of the main questions which could appear when nephrologists begin to use the HRQOL measures with renal patients: we clearly define the concept of HRQOL; we made recommendations about the instruments to evaluate HRQOL (generic vs specific instruments); and we show how to refine a HRQOL score to make it practical and comprehensible (the standardization of the scores by age and gender). PMID- 12090333 TI - Case report: primary retroperitoneal pelvic hydatid disease. PMID- 12090334 TI - Ureteral catheterization: getting the catheter through the nipple. PMID- 12090336 TI - Pediatric urolithiasis in Kuwait. AB - In this retrospective study, 31 Kuwaiti children with renal stones were reviewed between January 1996 and September 2000. Male to Female ratio was 2.1:1 with a mean age at presentation of 38 months. Family history of renal stones was reported in 58%. Stones were localized to the kidneys in 74.1%, to ureters in 6% and to the bladder in 9.6%. Bilateral stones were found in 64.5%. Clinical manifestations included: hematuria in 70.9%, passage of stones in 64.5%, abdominal pain in 41.9%, urinary tract infections in 29%, sterile pyuria in 22.9% and urine retention in 16%. Causes of stone formation included hypercalciuria in 38.7%, hyperoxaluria in 19.3%, cystinuria in 12.9%, xanthinuria in 12.9%, urinary tract infection in 3.2%, obstruction in 9.6% and idiopathic in 3.2%. In conclusion, lithogenic metabolic causes were found to be the major predisposing factors to stone formation among Kuwaiti children whereas diet and environmental factors played a trivial role. PMID- 12090335 TI - Current problems and needs in the treatment of pT1 G3 bladder carcinoma. PMID- 12090338 TI - The efficacy of neoadjuvant chemotherapy in invasive bladder cancer. AB - Radical cystectomy is the gold standard in the treatment of invasive bladder cancer. However, five-year disease-free survival is low most probably due to micrometastatic disease at the time of surgery. The neoadjuvant chemotherapy may be performed as the first line management for invasive bladder tumors in order to treat micrometastases found at the diagnosis and improve resectability of larger neoplasms. A total of 43 patients diagnosed with invasive bladder tumors and 11 patients received neoadjuvant chemotherapy. The mean age of patients was 64 (43 74) years, and mean follow-up period was 52 months (12-114). Neoadjuvant chemotherapy protocol consisted of methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC) or cisplatin, methotrexate, and cisplatin (CMV). All patients in neoadjuvant chemotherapy group underwent radical cystectomy. There was no significant difference between the groups with respect to disease-free survival time and overall survival time. In patients who received neoadjuvant chemotherapy, the respective disease-free and overall survival times were 31 months and 36 months versus 30 months and 35 months in patients who were treated with surgery only (p > 0.05). Five-year survival rates were 36% and 31% in the chemotherapy and no-chemotherapy groups, respectively. In the present study, 5 year survival rate was not affected by neoadjuvant chemotherapy in invasive bladder tumor. Complete pathological remission (stage p0) was found in 28% and pathological downstaging (stage < T2) was seen in 9% of patients in the neoadjuvant chemotherapy group. Five-year survival rates were 75% and 14.2% in patients who responded to chemotherapy, and in patients with no response, respectively (p < 0.05). The most favorable prognostic factor in this study was the response to neoadjuvant chemotherapy revealed as complete remission or pathological downstaging. The most important issue remains the prediction of patients who would respond and benefit from neoadjuvant chemotherapy. PMID- 12090337 TI - Multivariate evaluation of factors affecting recurrence, progression, and survival in patients with superficial bladder cancer treated with intravesical bacillus Calmette-Guerin (Tokyo 172 strain) therapy: significance of concomitant carcinoma in situ. AB - To evaluate the relative importance of clinicopathological factors affecting recurrence, progression, and survival in patients with superficial bladder cancer (pTa and pT1) undergoing bacillus Calmette-Guerin (BCG) therapy (Tokyo 172 strain), we reviewed data for 146 patients treated between 1985 and 1998. The median follow-up period was 64.7 months. Tumour recurrence, progression, and death were evaluated as endpoints using Cox's proportional hazards model. The 5 year recurrence-free rate was 56% for all 146 patients. Those with a past history of bladder cancer (n = 73) had significantly earlier recurrence than those without (n = 73, p = 0.017) and this tended to be the case for concomitant CIS (n = 34) although this did not reach statistical significance. The 5-year progression rate was 15% for all 146 patients and univariate analysis revealed that the presence of concomitant CIS was significantly associated with disease progression (p = 0.002). Multivariate analysis using the proportional hazards model confirmed the finding that only one factor, concomitant CIS, was significantly associated with progression. The 5-year survival rate was 84% for all 146 patients. Furthermore, univariate and multivariate analyses revealed that patient age, history of bladder cancer, and concomitant CIS were variables significantly related to patient survival. The present findings suggest that careful follow-up is mandatory after BCG instillation therapy for patients with superficial bladder cancer and concomitant CIS because of their relatively poor prognosis. PMID- 12090339 TI - Primary carcinoid tumor of the urinary bladder. AB - A polyp of about 1.0 cm diameter was incidentally found at the center of the posterior wall of the urinary bladder in a 73-year-old man on cystoscopic examination. A polyp was resected through a transurethral route without recurrence for 22 months. A polyp consisted of a tumor covered with mucosa of the bladder. Tumor cells have round or oval nuclei with fine chromatin and one or two nucleoli per nucleus, and finely granular eosinophilic cytoplasm, being arranged in a trabecular or tubular pattern. Furthermore, they showed positive staining for chromogranin-A and with Grimelius stain, indicating that the tumor is a carcinoid tumor. Since there was no evidence of carcinoid tumors in organs other than the urinary bladder, the present case is a primary carcinoid tumor (well differentiated endocrine tumor) of the urinary bladder which is extremely rare. PMID- 12090340 TI - The prognostic value of proliferating cell nuclear antigen, Ki-67 and nucleolar organizer region in transitional cell carcinoma of the bladder. AB - OBJECTIVES: To investigate the value of proliferating cell nuclear antigen (PCNA), Ki-67 antigen labelling indices and nucleolar organizer region (NOR) score in relation to histological grade, stage, recurrence and progression of the bladder tumor. MATERIALS AND METHODS: Tissue specimens from 77 bladder cancer patients (43 superficial, 34 invasive) were immunostained with PCNA and Ki-67 and stained with AgNOR. Thirteen specimens of normal bladder mucosa served as controls. RESULTS: In comparison to normal bladder mucosa the values of the three indicators were significantly greater (p < 0.001). There was a significant relationship between PCNA, Ki-67 indices, AgNOR scores and grade and stage of the tumor (p < 0.001). All indicators also correlated with each other (p < 0.001). The Kaplan-Meier curves for recurrence-progression free survival revealed that patients with a PCNA labelling index >36.22%, Ki-67 labelling index >29.68% and AgNOR score > 3.34 had a worse prognosis than those with <36.22%, <29.68% and <3.34, respectively. CONCLUSIONS: PCNA, Ki-67 indices and AgNOR scores correlated with each other and with tumor grade and stage. These proliferation markers may give objective and accurate information about the biological behavior of bladder transitional cell carcinoma. PMID- 12090341 TI - Update on the medical management of benign prostatic hyperplasia. PMID- 12090342 TI - Is there a difference in early perioperative morbidity in transurethral resection of prostate (TURP) versus TURP with cystolitholapaxy and TURP with inguinal herniorrhaphy? AB - Objective of this study is to determine the difference in early peri-operative morbidity of transurethral resection of prostate (TURP) and if it is combined with inguinal hernia repair and mechanical and/or pneumatic fragmentation of bladder calculus. All patients undergoing TURP, cystolitholapaxy (CLL), inguinal hernia repair (IHR) or any combination between January 1997 and December 1999 were identified using ICD 9CM coding and indexing system. Overall 1273 patients were identified, charts were reviewed for demographics, pre-operative parameter, intra-operative data and early peri-operative morbidity. In the three year period, 19 patients had TURP and inguinal herniorrhapy (IHR), 17 patients had TURP and cystolitholapaxy (CLL), 2 patients had TURP+IHR+CLL performed simultaneously; in the same period 346, 815, 74 patients had TURP, IHR, and CLL alone respectively. All the groups were matched for age, presentation and Co morbidities. There was significant difference in the operating time between the different groups. Complications were not significantly different in the TURP, IHR, CLL, TURP+IHR, and TURP+CLL. In conclusion, mean operative time for TURP+IHR is increased by a fraction of 168.3 for TURP+CLL by 109.8 and TURP+IHR+CLL is 202.1 over TURP alone, with no significant difference in morbidity between TURP and TURP+IHR and TURP+CLL. PMID- 12090343 TI - A comparison of two different incisional approaches in the surgical treatment of renal cell carcinomas. AB - Radical nephrectomy is the golden standard in the surgical treatment of renal cell carcinomas. In radical nephrectomy, the type of incision that is preferred is very important for the success of surgery. Within the last 6 years, we operated 76 renal cell carcinoma cases in our department. In these operations, we used either extra peritoneal flank incision (EFI) or transabdominal subcostal incision (TASI). In this study, we decided to compare the effects of these two incisional approaches on the clinical prognosis of the disease. We realized that TASI was our method of choice in tumors with high stage and large diameters. EFI on the other hand, was chosen for the cases with low stage. Both methods did not differ regarding intraoperative and postoperative complications. We concluded that in the treatment of renal cell carcinoma, the type of incision should be chosen according to the clinical picture of the disease and the experience of the surgeon in order for the operation to be successful. PMID- 12090345 TI - DNA flow-cytometric, histological and hormonal analysis of sertoli cell only syndrome (SECOS). AB - Sertoli cell only syndrome (SECOS) was identified on histology in 21 cases (16,28%) among 129 testicular biopsies performed in our department for azoospermia over the last 5 years. In these patients history, clinical features, hormonal levels, and histological findings were analyzed. In addition DNA flow cytometric analysis was performed and showed an almost complete absence of haploid cells. All patients presented with elevated serum FSH levels suggesting a Sertoli cell damage or reduced production of inhibin due to the absence of sermatogenic cells. An good correlation was found between histological findings and DNA histograms. In conclusion SECOS is a syndrome of unknown aetiology presenting in men with azoospermia. DNA flow-cytometric analysis is a reliable, rapid and easy method in the diagnosis of SECOS, and can replace histological examination. PMID- 12090344 TI - Comparison of T1c versus T2 prostate cancers in Japanese patients undergoing radical prostatectomy. AB - In order to examine the characteristics of patients with nonpalpable prostate cancer (T1c cancer) in Japan, patients treated with radical prostatectomy were compared with those with palpable (T2) cancer. Prostate-specific antigen (PSA) level in patients with T2b disease was significantly higher than those with T1c and T2a tumors. At the time of radical prostatectomy, 78%, 71% and 31% of patients with T1c, T2a, and T2b, respectively, had organ-confined disease. When insignificant cancer was defined as volume 0.5 ml or less and Gleason score less than 5, only 2 of 34 (5.9%) with clinical T1c disease were clinically insignificant. T1c cancers were clinically significant and clinicopathological features of Tlc tumors were similar to T2a tumors. PSA measurement could detect potentially curable prostate cancer. PMID- 12090347 TI - The effect of electromagnetic field on undescended testis after orchiopexy. AB - Undescended testis is a common problem leading to infertility. After orchiopexy some studies support the necessity of hormonal therapy. Electromagnetic field stimulation on living tissues increase cell proliferation, protein and DNA synthesis. Sixteen prepubertal rats was objected to the fixation of left testes to the anterior abdominal wall for 30 days, right testes were removed. Another group of sixteen rats objected only to the right orchiectomy and a manipulation simulating study group without fixation. After orchiopexy, animals were divided into two groups. Both groups had eight rats. Electromagnetic field (EMF) stimulation group had the stimulation for two hours every day for ten days, while the second group did not. The sham group also divided into two groups. The first one applied EMF and name as Group CEM, the second one was sham. Weight of removed testes were measured and fixed in 10% formaldehyde for histopathological evaluation. At the creating of undescended testis and right orchiectomies a blood sample was obtained for testosterone level of prepubertal rats. After finishing EMF stimulation the rats were mated with females for 17 days. After fertility study a blood sample was obtained for testosterone assay and body weight were measured and fixed in formaline for histopathologic evaluation. All the rats were killed with overdose ether anesthesia and number of fetuses were recorded. Histopathological evaluation was based on Johnsen criteria and seminiferous tubule diameter measurements. We conclude that EMF stimulation resulted in Leydig cell proliferation, increase in testosterone level, testis weight, but decrease in germ cell population. PMID- 12090346 TI - Pentoxifylline improves blood flow to both testes in testicular torsion. AB - OBJECTIVES: Electromagnetic and radioisotopic studies have shown that unilateral testicular torsion causes a decrease in contralateral testicular blood flow. Pentoxifylline improves microvascular blood flow in conditions of vascular insufficiency. An experimental study was designed to evaluate the effects of pentoxifylline (Ptx) on blood flow to both testes during unilateral testicular torsion and detorsion. MATERIALS AND METHODS: Thirty-six adult male albino Wistar rats were randomly divided into six groups where each consisted of six rats: group 1: sham operation, group 2: sham operation with Ptx, group 3: torsion, group 4: torsion with Ptx, group 5: detorsion, group 6: detorsion with Ptx. After intraperitoneal administration of Ptx at a dose of 50 mg/kg 15 minutes before torsion; right testes of the rats underwent 30 minutes of torsion and 30 minutes of detorsion. Blood flows of both testes were measured during torsion and detorsion simultaneously by using 133Xe clearance technique. RESULTS: Unilateral testicular torsion caused decrease in bilateral testicular blood flow. Pentoxifylline had no effect on testicular blood flow during torsion. Detorsion caused a partially increase in blood flow to ipsilateral (detorted) testis, but had no effect on contralateral (nontorted) testicular blood flow. Pentoxifylline administration during detorsion significantly increased blood flow to both testes. CONCLUSIONS: Testicular torsion is a pathological process that causes decreased blood flow to both testes. Pentoxifylline improves blood flow to both testes during detorsion in a rat model of testicular torsion. Further studies are needed to evaluate the effects of pentoxifylline on testicular torsion. PMID- 12090348 TI - The effects of female age on the outcome of testicular sperm extraction and intracytoplasmic sperm injection in infertile patients with azoospermia. AB - INTRODUCTION: Testicular sperm extraction (TESE) is well-defined procedure for surgical sperm retrieval in obstructive and non-obstructive azoospermia. This study was focused on the effectiveness of testicular sperm extraction and intracytoplasmic sperm injection (ICSI) for azoospermic men with different female age subgroups. MATERIALS AND METHODS: A total of 107 men with azoospermia underwent TESE and ICSI treatment. The women were examined in three groups 20-29, 30-34 and 35 years or older. The main outcome in this study was fertilization and pregnancy rates with TESE and ICSI. RESULTS: Spermatozoa were successfully retrieved during 97 of 107 (90.7%) TESE attempts, resulting in the fertilization of 286 of 563 (50.4%) injected metaphase II oocytes. Two hundred and fifty-five of them were transferred (89.8%). The clinical pregnancy rate and ongoing pregnancy rate per embryo transfer were 22.5% and 20.6% respectively. When comparing the fertilization and pregnancy rates, it was observed that women between the ages of 20-29 years had significantly higher pregnancy rates than women over 34 years of age (p < 0.05). CONCLUSION: The female age is a major factor in determining successful implantation in ICSI. PMID- 12090349 TI - Liposomal atp or NAD+ protects human endothelial cells from energy failure in a cell culture model of sepsis. AB - Sepsis depletes intracellular stores of ATP and NAD+, leading to cellular energy failure. Liposome encapsulation improves intracellular delivery of bulky, charged molecules and substrates susceptible to extracellular enzyme degradation. We hypothesized that treatments with liposome encapsulated ATP or NAD+ would protect human endothelial cells exposed to endotoxin (LPS) and interferon-gamma (IFN gamma) from energy failure. Liposomal ATP and NAD+ were prepared by a modification of the thin film method. Human endothelial cells were exposed to LPS 50 microg/ml and IFN-gamma 50 ng/ml for 72 hours, and liposomal ATP and NAD+ treatments were dosed at 0 and 24 hours. Energy state was determined by rate of mitochondrial respiration as measured by WST-1 assay. Mitochondrial respiration significantly decreased to 57% +/- 3 of control in LPS/IFN-gamma exposed cells after 72 hours. Liposomal ATP (200 microM) and NAD+ (100 microM) completely reversed this respiratory depression while empty liposomes, free ATP (200 microM). and free NAD+ (100 microM) did not. These results support the hypothesis that treatments with liposome encapsulated ATP or NAD+ protect human endothelial cells from energy failure in a cell culture model of sepsis and potentially may provide a novel therapy for use in clinical sepsis. PMID- 12090350 TI - The effects of nabumetone, a cyclooxygenase-2 inhibitor, on cisplatin-induced 5 hydroxytryptamine release from the isolated rat ileum. AB - In order to elucidate 5-HT release influenced by PGE2 in the background of the anticancer drug-induced emesis, the effect of nabumetone, a COX-2 inhibitor, on the release of 5-HT from the isolated rat ileum was investigated. PGE2 produced a concentration-dependent increase (10(-9) to 10 M) and decrease (10(-8) to 10(-6) M) in 5-HT release. Arachidonic acid also demonstrated a similar bell-shaped 5-HT release. The arachidonic acid-induced 5-HT release at 3 x 10(-6) M (313.04 +/- 25.90%) was significantly inhibited by the concomitant perfusion with BRL10720 (10(-6) M) (161.98 +/- 19.4%, p<0.01), an active metabolite of nabumetone, or indomethacin (3 x 10(-7) M)(190.01 +/- 16.19%, p<0.05). BRL10720 (10(-6) M)(428.57 +/- 51.72%, p<0.05) significantly inhibited the increase in 5-HT release induced by cisplatin (10(-6) M)(748.56 +/- 136.31%), suggesting that PGE2would be involved in cisplatin-induced 5-HT release. The increase in 5-HT release from the isolated ileum 72 hrs after cisplatin administration, in a delayed-emesis animal model, was significantly inhibited by the in vivo 3-day administration of nabumetone or BRL10720, but was not affected by the 3-day administration of dexamethasone. After 72 hours, however, the in vivo 3-days administration of nabumetone, BRL10720 or dexamethasone had no effect on the increase in ileal 5-HT levels induced by cisplatin. The use of COX-2 inhibitors to ameliorate delayed emesis induced by cisplatin-based anticancer chemotherapy has been proposed. On the other hand, there is a possibility that dexamethasone works through a mechanism other than 5-HT release in delayed emesis. PMID- 12090351 TI - Determination of a chemopreventive agent, Oltipraz, in rat plasma and urine by high-performance liquid chromatography. AB - A high-performance liquid chromatographic method was developed for the determination of a chemopreventive agent, Oltipraz, in rat plasma and urine. The sample preparation was simple; 2 volumes of acetonitrile were added to deproteinize the biological sample. A 50-microl aliquot of the supernatant was injected onto a C18 reversed-phase column. The mobile phase, acetonitrile : 0.5 mM ammonium acetate (55: 45, v/v for rat plasma and 45 : 55, v/v for rat urine), was run at a flow-rate of 1.5 ml/min. The column effluent was monitored using an ultraviolet detector set at 305 nm. The retention times for Oltipraz in rat plasma and urine were approximately 5.8 and 8.6 min, respectively. The detection limits of Oltipraz in rat plasma and urine were 20 and 50 ng/ml, respectively. The coefficients of variation of the assay (within-day and between-day) were generally low (below 4.65%) in concentration ranges from 0.02 (0.05) to 10 microg/ml for rat plasma and urine. No interference from endogenous substances was found. PMID- 12090352 TI - A comparative study of chemokine rantes detection using ELISA and Western blot. AB - Chemokines in biological sample are frequently found at very low level. Further, concentrations of chemokines which were measured in the same tissue detected with different methods often differ in literature. Therefore, RANTES concentrations were quantified by application of sandwich enzyme-linked immunosorbent assay (ELISA) and Western blot analyses. The sensitivity of ELISA was found to be much higher than that of Western blot. Additionally, the detection time differed considerably as well. For biological and biochemical characterization of chemokines it is essential to implement the optimal purification and detection techniques. With an understanding of the technical procedures and some pitfalls, chemokine detection can be applied more reliably. PMID- 12090353 TI - Changes of lipocalin type prostaglandin D-synthase in the seminal plasma of subfertile man. AB - It was proposed that lipocalin type prostaglandin D synthase (L-PGD-S), a bifunctional protein both synthesizing PGD2 and transporting retinoids and other lipophilic ligands, could be involved in the development and the maturation of sperm. In the present study, the seminal plasma (SP) of 59 adult males was analyzed by standard WHO methods and immunoblotting, using a monospecific polyclonal antibody directed against L-PGD-S. Briefly, aliquots of SP (2.5 microl), were fractionated by polyacrylamide electrophoresis in the presence of sodium dodecyl sulfate, the blots were stained and densitometrically analyzed. To obtain quantitative data, the aliquot of SP was selected within the linear part of the dose/band intensity curve and a proper quality control was analyzed in all blots to normalize the intensity of the bands of different experiments. A significant reduction (p<0.05) of the L-PGD-S levels was observed in severe oligozoospermic patients compared to normozoospermic subjects and a significant correlation between L-PGD-S levels and sperm concentration was found, as reported by other authors. Further studies are warranted to evaluate the possible diagnostic and pharmacological applications of these observations. PMID- 12090354 TI - Red cell and plasma antioxidant components and atherosclerosis in Japanese quail: a time-course study. AB - Our study aims to examine changes in red cell and plasma antioxidant components in relation to age and aortic lesion development in SUS Japanese quail during a 12-week period of dietary cholesterol supplementation. One hundred adult SUS males were divided into 5 treatment groups and were fed either a control or a cholesterol-supplemented (0.5% w/w) diet. Birds were sacrificed after 0, 4, 8 and 12 weeks on the diets and examined for plaque development and antioxidant components status. "Aging" was associated with increases in the activity of red cell SOD and GPx. Significant correlations among red cell GRd, GPx and SOD activities were found in old but not in younger adult birds. Plasma triglyceride levels increased, while plasma tocopherol level decreased with aging. With birds on the cholesterol diet, plaque score increased with time and regressed significantly but negatively on plasma cholesterol level at the initial stage of atherogenesis. Aortic triglycerides showed a drastic increase in the early stage of atherogenesis but returned to the pretreatment level during the late stage. Conversely, aortic cholesterol showed small increases at the early stages but large increases during the late stage. Red cell antioxidant components showed increases at the early and late stages with a leveling off at the mid stage. Plasma GRd activity decreased while plasma tocopherol level increased (after adjusting for the effect of effect of aging) with cholesterol feeding. We conclude that the increase in plasma triglyceride levels and associations among red blood cell GRd, GPx and SOD activities in "old" birds fed the control diet resembled the situation in the early stages of atherogenesis in the cholesterol fed birds. This would be consistent with the known permissive effect of aging on the course of atherogenesis. Triglycerides may play a crucial role in atherogenesis during the early phase of lesion development. Early and late phases of lesion development are biochemically distinct, indicating that the process of atherogenesis is a highly dynamic one. The patterns of antioxidant alterations associated with lesion development showed a complex time-dependence. PMID- 12090355 TI - Prior treatment with captopril attenuates carbon tetrachloride-induced liver injury in mice. AB - The present investigation focused on the possible hepatoprotective potential of captopril on carbon tetrachloride (CCl4)-induced acute liver injury in mice. Twenty-four hours after a single intraperitoneal injection of CCl4 (20 microl/Kg), hepatotoxicity was evidenced in the serum by elevated levels of aspartate transaminase (AST; EC: 2.6.1.1), alanine transaminase (ALT; EC: 2.6.1.2) and lactate dehydrogenase (LDH; EC: 1.1.1.27) and in the liver by depleted level of reduced glutathione (GSH), enhanced activity of glutathione peroxidase (GSH-Px; EC: 1I.11.1.9) and elevated level of lipid peroxides (LP). Captopril was given orally at three dose levels viz., 10, 25 and 50 mg/Kg/day for three consecutive days before subjecting the animals to the hepatotoxin. With the exception of the lowest dose namely, 10 mg/Kg/day, captopril afforded protection against CCl4-induced hepatotoxicity to different extents. Thus, the elevated activities of the enzymes AST, ALT, LDH and GSH-Px as well as the enhanced lipid peroxidation were markedly reduced below those elicited by the hepatotoxin, reaching values closer to the control, though still statistically higher. Captopril, however, did not ameliorate the depletion of GSH produced by CCl4. The data reported herein reveal a protective potential of captopril against the acute hepatotoxicity induced by CCl4 in mice. This hepatoprotection could be attributed, at least in part, to the free radical scavenging properties of the drug. PMID- 12090356 TI - Correlation of circulating interleukin-10 with thyroid hormone in acute myocardial infarction. AB - In patients with acute myocardial infarction (AMI), euthyroid sick syndrome (ESS) has been reported to be linked to increase in interleukin (IL)-6 and activation of its receptors. Recent reports have shown that IL-10, an anti-inflammatory cytokine, also plays a key role in the pathogenesis of AMI. Therefore we investigated the relationship between thyroid state and IL-10 in patients with AMI. We measured thyrotropin (TSH), free triiodothyronine (FT3), free thyroxine (FT4), IL-10, and IL-6 in plasma from 20 patients with AMI and 20 healthy controls. All 20 AMI patients showed significantly lower concentrations of FT3 than in controls accompanied by normal or subnormal levels of TSH, characterized ESS. Concentrations of IL- 10 and IL-6 were higher in patients than in controls. Both IL-10 and IL-6 significantly (p<0.05, respectively) correlated with thyroid hormone in patients with AMI. Time course of IL-10, IL-6, and FT3 seemed to be tightly linking. In conclusion, IL-10 and IL-6 appears to affect thyroid hormone homeostasis in patients with AMI. PMID- 12090357 TI - Alpha1-adrenergic receptors and their significance to chemical-induced nephrotoxicity--a brief review. AB - Stimulation of alpha-adrenergic receptors by cold stress or adrenergic agents has been shown to potentiate the toxicity of numerous toxicants. Several lines of evidence indicate that this interaction is dependent on glutathione depression and increased cytosolic Ca2+ concentrations produced by alpha1-adrenergic compounds. In this report, evidence is provided in support of the mechanism of adrenoreceptor-mediated potentiation of nephrotoxicity. Alpha1-adrenergic agonists are shown to potentiate the toxicity of nephrotoxicants that exert their toxic effects via glutathione conjugation or Ca2+ deregulation. This review summarizes the effects of the alpha1-adrenergic agonist, phenylephrine, at enhancing the toxicity of 2-bromohydroquinone, 1,2-dibromoethane, and cis diammineplatinum(II) dichloride. PMID- 12090358 TI - Hepatoprotective ability of a novel botanical formulation on mild liver injury in rats produced by acute acetaminophen and/or alcohol ingestion. AB - Medicinal herbs have been used for centuries in an attempt to overcome hepatic dysfunctions emanating from ingestion of hepatotoxic substances. However, the vast majority of information concerning their use is anecdotal. Well-performed animal studies would lend credence to the concept that some medicinal herbs may prevent or, at least ameliorate, hepatic dysfunction arising from drug-induced toxicity. The present investigation examined the potential for a combination of medicinal herbs to favorably influence the course of mild/moderate acute hepatic injury induced in rats by the oral intake of acetaminophen and/or ethyl alcohol. We performed four separate studies using elevations of liver enzymes [aspartate aminotransferase (AST) and alanine aminotransferase (ALT)] as our primary biomarkers of hepatotoxicity. In the first study, the ability of orally administered acetaminophen at different doses to produce acute hepatotoxicity was examined. In the second and third studies, the ability of a combination of medicinal herbs (a novel botanical formulation) was assessed to ameliorate the acetaminophen-induced hepatotoxicity. In the last series of studies, we used acute alcohol ingestion to cause liver perturbations and examined the ability of medicinal herbs to overcome hepatotoxicity. We also evaluated the ability of the medicinal herb combination to overcome acetaminophen-induced hepatotoxicity in rats simultaneously challenged with ethyl alcohol ingestion. Hepatotoxicity, estimated by increased levels of AST and ALT, was produced by a 2.0 mg/Kg oral dose of acetaminophen but not by lesser doses. Treatment with a combination of medicinal herbs (a novel botanical formulation) significantly ameliorated acetaminophen-induced toxic response. The combination of medicinal herbs also decreased the hepatic toxicity produced by acute ethyl alcohol ingestion. We conclude that oral ingestion of a novel botanical formulation (a combination of medicinal herbs) is effective in lessening drug-induced hepatotoxicity produced by acetaminophen and/or ethyl alcohol in an animal model. PMID- 12090359 TI - Levels of lipid peroxidation products in a chronic inflammatory disorder. AB - We have examined the plasma levels of the lipid peroxidation products 4-hydroxy nonenal and malondialdehyde in a carefully controlled study of age and sex matched subjects with rheumatoid arthritis in whom potentially confounding influences such as disease modifying anti-rheumatic drugs (DMARDs), self medication and vitamin supplements were eliminated. The plasma concentrations of the antioxidants uric acid and vitamin E were also measured. The results reveal a strong and consistent inverse correlation between the levels of lipid peroxidation products in the plasma and erythrocyte sedimentation rate (ESR). There is no indication that vitamin E or urate function as the major antioxidant agent in arthritis, as has been suggested in more seriously affected patients. It is concluded that there is no evidence that vitamin E is more important, and urate less important as an antioxidant in mild arthritis. The correlation between lipid peroxidation and ESR suggests a more complex relationship than has been assumed. PMID- 12090360 TI - Effects of polyethyleneglycol-superoxide dismutase (PEG-SOD) and pentoxifylline on small intestinal anastomoses established in the 24th hour of reperfusion: an experimental study in rats. AB - In a previous study, we had shown that anastomoses established in the 24th hour of reperfusion healed less than the ones established in earlier periods. In this study, we aimed to assess the impacts of polyethyleneglycol-superoxide dismutase (PEG-SOD), a free oxygen radical scavenger and of pentoxyfilline, a methyl xanthine derivative on anastomotic healing at 24th hour of reperfusion. 18 Wistar albino rats were divided into 3 groups (n = 6). In all groups superior mesenteric artery was occluded for 40 minutes and the intestine was reperfused for 24 hours. Rats were relaparotomized in the 24th hour and small intestinal anastomoses were established. In Sham group, saline (0.5ml); in Group PTX, pentoxyfilline (25mg/kg); and in Group PGS, PEG-SOD (1,500U/kg) was administered intravenously 20 minutes before anastomoses. In the 5th day, anastomotic healing was evaluated by bursting pressures and hydroxyproline contents. Results were analized by Mann Whitney U test, p < 0.05 was considered significant. Both of average bursting pressures and average hydroxyproline contents were highest in Group PGS (240 mmHg +/- 23.9; 7.71 +/- 0.68 micromol/g-tissue), followed by sham group (168.3 +/- 18.3 mmHg; 5.94 +/- 0.62 micromol/g-tissue) and Group PTX (83.8 +/- 9.2 mmHg, 5.94 +/- 0.62 micromol/g-tissue). Differences of these two parameters between all groups were statistically significant (p < 0.05). Best anastomotic healing in the 24th hour of reperfusion was achieved by PEG-SOD administration, whereas pentoxyfilline did not favor the healing. PMID- 12090362 TI - Behavioral medicine and clinical health psychology: introduction to the special issue, a view from the decade of behavior. AB - This collection of articles follows prior special issues on behavioral medicine published in the Journal of Consulting and Clinical Psychology in 1982 and 1992. From the vantage point of the Decade of Behavior, the field has grown in scope, depth of basic science foundations, and evidence of applied utility. Yet many challenges remain-especially in addressing a wide range of health problems across diverse populations and in a health care context characterized by rapid changes in technology and by a growing concern with costs and evidence-based practice. PMID- 12090363 TI - Sociodemographic diversity and behavioral medicine. AB - The broad array of economic and cultural diversity in the U.S. population correlates with and impacts on the study of behavioral aspects of health. The purpose of this article was to provide a selective overview of behavioral medicine research on sociodemographically diverse populations, with a focus on ethnicity, gender, and socioeconomic status. Suggestions are provided with regard to methodological refinement of research and insights into possible future directions in behavioral medicine research on ethnically and economically diverse populations. PMID- 12090361 TI - 2001 USPHS/IDSA guidelines for the prevention of opportunistic infections in persons infected with human immunodeficiency virus. PMID- 12090365 TI - Smoking cessation: progress, priorities, and prospectus. AB - The authors review developments in understanding smoking cessation interventions over the past decade. Noteworthy is the unprecedented growth of research and knowledge that has left a deeper understanding of how best to use new and existing behavioral and pharmacologic tools and strategies to help smokers quit. The status of public-health-level interventions is evaluated, questions are raised concerning their efficacy, and suggestions are offered for further refinement of these intervention strategies. Development of cessation guidelines is reviewed, and the state of knowledge concerning behavioral and pharmacologic interventions is summarized. The authors also present agendas for behavioral and pharmacologic research related to smoking cessation and discuss individual difference factors among smokers that may prove to be important in designing new and refining existing treatments. PMID- 12090364 TI - Applications of cost-effectiveness methodologies in behavioral medicine. AB - In 1996, the Panel on Cost-Effectiveness in Health and Medicine developed standards for cost-effectiveness analysis. The standards include the use of a societal perspective, that treatments be evaluated in comparison with the best available alternative (rather than with no care at all), and that health benefits be expressed in standardized units. Guidelines for cost accounting were also offered. Among 24,562 references on cost-effectiveness in Medline between 1995 and 2000, only a handful were relevant to behavioral medicine. Only 19 studies published between 1983 and 2000 met criteria for further evaluation. Among analyses that were reported, only 2 studies were found consistent with the Panel's criteria for high-quality analyses, although more recent studies were more likely to meet methodological standards. There are substantial opportunities to advance behavioral medicine by performing standardized cost-effectiveness analyses. PMID- 12090366 TI - Obesity: responding to the global epidemic. AB - Obesity has reached epidemic proportions in the United States and other developed nations. In the United States, 27% of adults are obese and an additional 34% are overweight. Research in the past decade has shown that genetic influences clearly predispose some individuals to obesity. The marked increase in prevalence, however, appears to be attributable to a toxic environment that implicitly discourages physical activity while explicitly encouraging the consumption of supersized portions of high-fat, high-sugar foods. Management of the obesity epidemic will require a two-pronged approach. First, better treatments, including behavioral, pharmacologic, and surgical interventions, are needed for individuals who are already obese. The second and potentially more promising approach is to prevent the development of obesity by tackling the toxic environment. This will require bold public policy initiatives such as regulating food advertising directed at children. The authors call not for the adoption of a specific policy initiative, but instead propose that policy research, based on viewing obesity as a public health problem, become a central focus of research. PMID- 12090367 TI - Physical activity and exercise: recent advances and current challenges. AB - This article presents an overview of major developments in physical activity and exercise for health promotion and disease prevention during the past 10 years. The importance of physical activity to physical and mental health was increasingly recognized. Assessment methods appropriate for diverse population groups were developed, and understanding of correlates of regular physical activity improved. Many studies focused on moderate activity that can be integrated into people's everyday lives. Research in community, school, and health care settings demonstrated that interventions based on cognitive behavioral theories can change physical activity. Nevertheless, physical activity remained at low levels in most population subgroups throughout the past decade. Psychologists can make important contributions to efforts to address this ongoing challenge to public health. PMID- 12090368 TI - Psychoneuroimmunology: psychological influences on immune function and health. AB - This review focuses on human psychoneuroimmunology studies published in the past decade. Issues discussed include the routes through which psychological factors influence immune function, how a stressor's duration may influence the changes observed, individual difference variables, the ability of interventions to modulate immune function, and the health consequences of psychosocially mediated immune dysregulation. The importance of negative affect and supportive personal relationships are highlighted. Recent data suggest that immune dysregulation may be one core mechanism for a spectrum of conditions associated with aging, including cardiovascular disease, osteoporosis, arthritis, Type 2 diabetes, certain cancers, and frailty and functional decline; production of proinflammatory cytokines that influence these and other conditions can be stimulated directly by negative emotions and indirectly by prolonged infection. PMID- 12090369 TI - Psychosocial influences on the development and course of coronary heart disease: current status and implications for research and practice. AB - Psychosocial characteristics predict the development and course of coronary heart disease (CHD). In this review, the authors discussed human and animal research on psychophysiological mechanisms influencing coronary artery disease and its progression to CHD. They then reviewed literature on personality and characteristics of the social environment as risk factors for CHD. Hostility confers increased risk, and a group of risk factors involving depression and anxiety may be especially important following myocardial infarction. Social isolation, interpersonal conflict, and job stress confer increased risk. Psychosocial interventions may have beneficial effects on CHD morbidity and mortality, although inconsistent results and a variety of methodological limitations preclude firm conclusions. Finally, they discussed implications for clinical care and the agenda for future research. PMID- 12090370 TI - Biobehavioral approaches to the treatment of essential hypertension. AB - Despite recent advances in the medical management of hypertension, chronically elevated blood pressure remains a major health problem in the United States, affecting almost 50 million Americans. It is widely recognized that lifestyle factors contribute to the development and maintenance of elevated blood pressure. This article critically reviews current approaches to the nonpharmacological treatment of high blood pressure and highlights outcome studies of exercise, weight loss and dietary modification, and stress management and relaxation therapies. Methodological issues in the assessment and treatment of hypertension are discussed, along with possible mechanisms by which lifestyle modification may reduce elevated blood pressure. PMID- 12090372 TI - Diabetes and behavioral medicine: the second decade. AB - Diabetes management depends almost entirely on behavioral self-regulation. Behavioral scientists have continued a collaboration with other health systems researchers to develop a holistic approach to this disease. The authors summarized the literature in 4 major areas: self-management of diabetes, psychosocial adjustment and quality of life, neuropsychological impact, and psychobehavioral intervention development. Progress made in each of these areas over the past decade is highlighted, as are important issues that have not yet received sufficient scientific attention. Emerging areas likely to become central in behavioral research, such as diabetes prevention, are introduced. The future of behavioral medicine in diabetes is also discussed, including topics such as the changing role of psychologists in diabetes care, the urgent need for more and better intervention research, the growing importance of incorporating a health system-public health perspective, and obstacles to the integration of psychobehavioral approaches into routine health care delivery. PMID- 12090373 TI - Behavioral research in HIV/AIDS primary and secondary prevention: recent advances and future directions. AB - Great advances have been made over the past decade in behavioral research on how to help persons avoid contracting HIV infections (primary prevention) and how to reduce or alleviate adverse consequences among persons who are living with HIV disease (secondary prevention). Within the primary prevention areas, research has shown the effectiveness of risk-reduction interventions undertaken with individuals, couples, small groups, communities, and at a social policy/structural level. Advances in HIV medical care have also created important new challenges and roles for behavioral scientists in the area of HIV secondary prevention. This article concludes by identifying key emerging issues in HIV behavioral research that will require attention in the years ahead. PMID- 12090371 TI - Biobehavioral outcomes following psychological interventions for cancer patients. AB - Psychological interventions for adult cancer patients have primarily focused on reducing stress and enhancing quality of life. However, there has been expanded focus on biobehavioral outcomes--health behaviors, compliance, biologic responses, and disease outcomes--consistent with the Biobehavioral Model of cancer stress and disease course. The author reviewed this expanded focus in quasiexperimental and experimental studies of psychological interventions, provided methodologic detail, summarized findings, and highlighted novel contributions. A final section discussed methodologic issues, research directions, and challenges for the coming decade. PMID- 12090374 TI - Recent advances and future directions in the biopsychosocial assessment and treatment of arthritis. AB - This article provides an overview of the emerging literature on biopsychosocial assessment and treatment for two of the most common forms of arthritis: osteoarthritis and rheumatoid arthritis. The article is divided into 3 parts. In the 1st part, the basic elements of the biopsychosocial approach to assessing and treating persons having arthritis is described. In the 2nd part, the authors evaluate studies of biopsychosocial approaches to the assessment of arthritis pain and disability. Six research areas are reviewed: learned helplessness, depression, stress, pain coping, self-efficacy, and the social context of arthritis. The 3rd part of the article reviews studies that testing the efficacy of biopsychosocial treatment approaches for persons having osteoarthritis and rheumatoid arthritis. PMID- 12090375 TI - Assessment and psychological management of recurrent headache disorders. AB - This article updates earlier reviews of recurrent headache disorders published in 1982 and 1992, selectively reviewing research published since 1990. Current issues in assessment (headache diagnosis, psychophysiology, comorbid psychopathology, quality-of-life assessment, and new assessment technologies) and psychological treatment (efficacy, therapeutic mechanisms, treatment delivery, and integration with drug therapy) are addressed. The author emphasizes the need to adapt psychological treatments to the severity of the headache disorder and to developments in drug therapy. Opportunities for the integration of biological, medical, and psychological science are highlighted. PMID- 12090376 TI - Psychological factors in chronic pain: evolution and revolution. AB - Research has demonstrated the importance of psychological factors in coping, quality of life, and disability in chronic pain. Furthermore, the contributions of psychology in the effectiveness of treatment of chronic pain patients have received empirical support. The authors describe a biopsychosocial model of chronic pain and provide an update on research implicating the importance of people's appraisals of their symptoms, their ability to self-manage pain and related problems, and their fears about pain and injury that motivate efforts to avoid exacerbation of symptoms and further injury or reinjury. They provide a selected review to illustrate treatment outcome research, methodological issues, practical, and clinical issues to identify promising directions. Although there remain obstacles, there are also opportunities for psychologists to contribute to improved understanding of pain and treatment of people who suffer from chronic pain. The authors conclude by noting that pain has received a tremendous amount of attention culminating in the passage of a law by the U.S. Congress designating the period 2001-2011 as the "The Decade of Pain Control and Research." PMID- 12090377 TI - Psychological aspects of asthma. AB - Asthma can be affected by stress, anxiety, sadness, and suggestion, as well as by environmental irritants or allergens, exercise, and infection. It also is associated with an elevated prevalence of anxiety and depressive disorders. Asthma and these psychological states and traits may mutually potentiate each other through direct psychophysiological mediation, nonadherence to medical regimen, exposure to asthma triggers, and inaccuracy of asthma symptom perception. Defensiveness is associated with inaccurate perception of airway resistance and stress-related bronchoconstriction. Asthma education programs that teach about the nature of the disease, medications, and trigger avoidance tend to reduce asthma morbidity. Other promising psychological interventions as adjuncts to medical treatment include training in symptom perception, stress management, hypnosis, yoga, and several biofeedback procedures. PMID- 12090378 TI - Psychological factors in end-stage renal disease: an emerging context for behavioral medicine research. AB - End-stage renal disease (ESRD) is a chronic, life-threatening condition afflicting over 300,000 Americans. Patient nonadherence and psychological distress are highly prevalent among ESRD patients, and both have been found to contribute to greater morbidity and earlier mortality in this population. A range of factors have been examined as potential determinants of adherence and adjustment. Evidence suggests that adherence and adjustment are maximized when a patient's preferred style of coping is consistent with the contextual features or demands of the renal intervention the patient is undergoing. Challenges for future clinical research include refining methodologies for the assessment of depression and adherence, more clearly evaluating the efficacy of psychological interventions, and clarifying the role that depression and social support play in influencing patient mortality. PMID- 12090379 TI - Psychosocial aspects of assessment and treatment of irritable bowel syndrome in adults and recurrent abdominal pain in children. AB - This article presents a selective review of psychosocial research on irritable bowel syndrome (IBS) in adults and on a possible developmental precursor, recurrent abdominal pain (RAP), in children. For IBS the authors provide a summary of epidemiology, of the psychological and psychiatric disturbances frequently found among IBS patients, and of the possible role of early abuse in IBS. A review of the psychosocial treatments for IBS finds strong evidence to support the efficacy of hypnotherapy, cognitive therapy, and brief psychodynamic psychotherapy. The research relating RAP to IBS is briefly reviewed, as is the research on its psychological treatment. Cognitive-behavioral therapy that combines operant elements and stress management has the strongest support as a treatment for RAP. PMID- 12090380 TI - Psychosocial factors in peptic ulcer and inflammatory bowel disease. AB - Over the past decade, while gastroenterologists' interest in mind-body interactions in organic disorders dwindled, stronger evidence has linked psychosocial factors with the incidence and recurrence of peptic ulcer and with the course of inflammatory bowel disease. Psychological-behavioral approaches to treatment continue to be disappointing. Psychosocial factors may affect ulcer by increasing duodenal acid load, altering local circulation or motility, intensifying Helicobacter pylori infection, stimulating corticosteroid secretion, and affecting health risk behaviors; possible mechanisms for inflammatory bowel disease include immune deregulation, gut permeability changes, and poor medication adherence. Both belong to the growing category of diseases thought to have an infectious component: for peptic ulcer the bacterium Helicobacter pylori, for inflammatory bowel disease an exaggerated immune response to gut bacteria. Peptic ulcer and inflammatory bowel disease, which present unique interactions among psychological, immunologic, endocrine, infectious, and behavioral factors, are splendid paradigms of the biopsychosocial model. PMID- 12090381 TI - Psychosocial aspects of selected issues in women's reproductive health: current status and future directions. AB - Emphasizing research published in the past decade, this article presents a summary and evaluation of psychosocial investigations of women's reproductive health, with a focus on selected aspects of menstruation, pregnancy and birth, infertility, and menopause. In some areas, studies have focused on negative physical and psychological concomitants of these health issues. However, research reveals substantial individual variability, with most women adapting well to reproductive health changes. Although methodological and conceptual shortcomings have limited firm conclusions, research has advanced our understanding of the multivariate biological, psychological, and social influences on women's reproductive health and associated outcomes. Understanding and promoting women's reproductive health across the lifespan requires biopsychosocial approaches to research. PMID- 12090382 TI - Psychological assessment and care of organ transplant patients. AB - Organ transplant has been developed in animal models over the past 100 years. The major limiting factor in transplant medicine is the shortage of donor organs. This shortage creates pressure for fair and efficient allocation of organs, with expectations that those involved in transplantation will strive to achieve optimal outcomes and ensure just access. This article reviews the major types of transplants and the illnesses and behavioral comorbidities that lead to these procedures, the psychological assessment of transplant candidates, the adaptive tasks required of the transplant recipient at various stages of the transplant process, and relevant psychological interventions. Liaison with others on the transplant team and ethical issues of concern to psychologists who work with transplant patients, including living organ donors, are also discussed. Finally, new developments in transplant and suggestions for future psychological research in organ transplant are presented. PMID- 12090383 TI - Genetic testing: psychological aspects and implications. AB - As the number of genes associated with inherited disease continues to grow, researchers and practitioners in behavioral medicine will encounter complex psychological issues faced by individuals at risk for these diseases. A review of the literature concerning prenatal, carrier, and predictive genetic testing suggests that the severity of psychological risks posed by research-based genetic testing is not great. However, subgroups of individuals with particular psychological traits may be more vulnerable to adverse effects. Available data do not provide evidence that genetic testing promotes changes in health-related behaviors. Thus, although there may be less of a role for mental health professionals in the psychological counseling of genetic testing participants, there is a need for research and practice to facilitate health protective behaviors in response to genetic risk information. PMID- 12090384 TI - Emotional disorders in primary care. AB - Individuals with emotional disorders are more likely to use primary medical care than specialty mental health services, but these disorders are likely to be undetected or inadequately treated. Recognition of the importance of primary medical care for the treatment of mental disorder has resulted in pressing new research priorities. One set of issues concerns the adequacy of existing nosological systems for conceptualizing emotional disorder in primary care and identifying need for treatment. Another concerns the difficulties translating efficacious treatment into effective strategies that can be integrated into the competing demands of primary medical care. Psychologists have played only a limited role in defining and addressing emerging questions. Irreversible changes in mental health services have created the need for the development of a psychosocial perspective for what would otherwise be defined as narrowly biomedical issues. PMID- 12090385 TI - Behavioral medicine approaches to somatoform disorders. AB - This article reviews the research evidence for the efficacy of cognitive behavioral therapy (CBT) for somatoform disorders. Randomized controlled studies support the efficacy of individual CBT for the treatment of hypochondriasis, body dysmorphic disorder (BDD), and undifferentiated somatoform disorders including medically unexplained symptoms, chronic fatigue syndrome, and noncardiac chest pain, and group CBT for the treatment of BDD and somatization disorder. On the basis of this review of the existing research and a theoretical model of the processes involved in somatoform disorders, the authors offer suggestions for future research and effective treatment. PMID- 12090386 TI - Adolescent health psychology. AB - In this article, a biopsychosocial model of adolescent development is used as an organizing framework for a review of primary, secondary, and tertiary prevention research with adolescent populations. During adolescence many critical health behaviors emerge, affecting future disease outcomes in adulthood. In addition, most of the predominant causes of morbidity and mortality in adolescence are unique to this period of development, indicating that health-focused interventions must be tailored specifically to adolescents. Moreover, it is during adolescence that lifelong patterns of self-management of and adjustment to chronic health conditions are established. Thus, an increased focus on adolescence in health psychology research is important both to improve the health of adolescents per se and to optimize health trajectories into adulthood. PMID- 12090387 TI - Behavioral medicine and aging. AB - A brief history of behavioral medicine and aging is followed by a series of perspectives that help to understand how age is used as a variable in this research, the relative importance of age to declines in cognitive functioning, and the impact of behavioral risk indicators on healthy survival. The authors discuss Alzheimer's disease and the role of age in clinical practice. Also discussed are potential problems in age-related research, ways to improve the knowledge base in behavioral medicine and aging, and thoughts about future challenges to enhance work in behavioral medicine and aging. PMID- 12090388 TI - Behavioral medicine: 2002 and beyond. AB - This article highlights future directions for research and practice in behavioral medicine. Topics addressed include social and environmental issues, the role of technology, translational research, improving and developing interventions, and professional training and retraining. PMID- 12090389 TI - Investigation of a cyclopic, human, term fetus by use of magnetic resonance imaging (MRI). AB - Using magnetic resonance imaging (MRI), the internal neural and craniofacial malformations of a cyclopic fetus are described. External facial features were characterized by a tubular proboscis situated above a single eye slit. The brain was recognized as 'pancake' type alobar holoprosencephaly (a condition where the undifferentiated telencephalon partially surrounds a monoventricle). Displacement of some bones that normally contribute to the orbit could be clearly discerned. Absence of neural structures (e.g. falx cerebri, corpus callosum) and missing components of the ethmoid bone indicated a midline deficit. This correlates with proposed theories of cyclopic embryopathy, which suggest that the prechordal plate and the neural crest cells are affected during the third week of gestation in cyclopia. PMID- 12090390 TI - Three-dimensional sonographic evaluation of the fetal lumbar spinal canal. AB - In a prospective cross-sectional ultrasound study the size of the fetal lumbar spinal canal was evaluated to determine reference values for the lumbar part of the vertebral canal. One hundred and sixty-seven pregnant women undergoing routine obstetric ultrasound were studied between 16 and 41 weeks of gestation. Exclusion criteria consisted of structural fetal anomalies or growth restriction. Area and volume of the vertebral canal at L1, L3 and L5 were calculated by three dimensional (3D) ultrasound. Length of the lumbar spine was also determined. The size of the spinal canal and spinal length correlated well with gestational age. No gestational-age-dependent differences in area and volume measurements between upper and lower lumbar spine were found. The results provide an in vivo assessment of the spinal canal by 3D ultrasound over the entire gestation period. PMID- 12090391 TI - An investigation of thoracic and lumbar cancellous vertebral architecture using power-spectral analysis of plain radiographs. AB - The internal architecture of the vertebral bodies spanning the levels T1 to L5 in seven male columns was studied using mammographic-resolution radiographs of 2.5 mm-thick planar parasagittal slices. The overlapping radiographic shadows of vertebral trabeculae combined in the image to form a series of 'elements', broadly representative of the cancellous structure. The orientations and sizes of these elements were analysed by applying the Fast Fourier transform (FFT) to the digitized radiographic images. Elements aligned in the 'vertical' orientation, along the long axis of the column, were the most prominent for all vertebral levels. The relative prominence of horizontal to vertical elements was generally constant along the column below T5. In contrast, the relative prominence of oblique to vertical elements declined in the cranio-caudal direction, particularly in individuals aged > or = 60 years. The ratio of 'large' (x > 0.3 mm) to 'small' (0.15 mm < or = x < or = 0.3 mm) elements was unchanged cranio caudally in specimens < 60 years. However, in individuals > or = 60 years, large elements increased in relative prominence in the caudal direction. These results suggest that a basic orthogonal pattern of trabeculae is found along the male human spine, regardless of differences in vertebral body size. Power-spectral analysis is shown to yield information summarizing the predominant orientations and sizes of radiographically rendered architectural elements of vertebral cancellous bone, to define the effects of ageing on architecture, and to identify broad structural differences between vertebral levels in the adult male spine. PMID- 12090393 TI - Myoarchitecture and vasculature of the heart ventricle in some freshwater teleosts. AB - The morphological characteristics of the ventricular myocardium and of coronary vascularization were studied in three freshwater teleost species, Piaractus mesopotamicus, Colossoma macropomum and Clarias gariepinus (African catfish), by correlating their ventricular shapes and swimming habits. In Piaractus mesopotamicus and Colossoma macropomum, species with highly active swimming habits, the cardiac ventricle showed a pyramidal shape and a richly vascularized myocardium consisting of an outer compact layer and inner spongy layer. In Clarias gariepinus, a less active species, we observed a saccular ventricle with a mixed myocardium and coronary arteries, in contrast to the ventricular structure of other species described in the literature. PMID- 12090392 TI - The functional anatomy of the human anterior talofibular ligament in relation to ankle sprains. AB - The anterior talofibular ligament is the most commonly injured ligament in the ankle. Despite considerable interest in the clinical outcome of treatment protocols, we do not know whether the distinctive pattern of localization of the injuries relates to regional differences in the structure and molecular composition of the ligament. To address this issue, ligaments were examined by histology and immunohistochemistry. Differences in the structure of its two attachments (i.e. entheses) were evaluated with quantitative, morphometric techniques, and regional differences in the distribution of collagens, glycosaminoglycans and proteoglycans were determined qualitatively by immunolabelling. Morphometric analyses showed that bone density was less at the fibular attachment, but that enthesis fibrocartilage was more prominent. Immunohistochemistry revealed the presence of a fibrocartilage (containing type II collagen and aggrecan) at the site where the ligament wraps around the lateral talar articular cartilage in a plantarflexed and inverted foot: the fibrocartilage is regarded as an adaptation to resisting compression. We propose that avulsion fractures are less common at the talar end of the ligament because (1) bone density is greater here than at the fibular enthesis, and (2) stress is dissipated away from the talar enthesis by the 'wrap-around' fibrocartilaginous character of the ligament near the talar articular facet. PMID- 12090394 TI - Immunohistochemical localization of BDNF-, TrkB- and TrkA-like proteins in the teleost lateral line system. AB - The lateral line system, formed of both superficial (pit organs) and canal neuromasts, is one of the major mechanosensory systems in fish. It has always been assumed that this system depends on neurotrophins and their cognate Trk receptors for development and maintenance, as has been shown in other mechanosensitive systems of vertebrates. However, until nowthis issue has not been specifically addressed. In this study we used immunohistochemistry to investigate the occurrence and localization both of neurotrophins (NGF-, BDNF- and NT-3-like) and of Trk-like proteins (TrkA-, TrkB-, TrkC-like) in alevins of Salmo salar and S. trutta. All cells in the pit organs of S. salar displayed strong immunoreactivity for TrkB-like and BDNF-like, whereas they were restricted to the hair cells in S. trutta. The hair, supporting and mantle cells of S. salar, and the mantle cells of S. trutta, also expressed TrkA-like immunoreactivity. In the canal neuromasts BDNF-, TrkA- and TrkB-like proteins were present in all cells, without differences between species. NGF-, NT-3- and TrkC-like immunoreactivity were never detected. The present results suggest that mechanoreceptive hair cells, as well as supporting cells, in the lateral line system are under the control of the BDNF-TrkB-like complex, and probably of ligands of TrkA-like receptors. PMID- 12090395 TI - Calcium-sensing receptors and parathyroid hormone-related protein in the caudal neurosecretory system of the flounder (Platichthys flesus). AB - The caudal neurosecretory system of the flounder (Platichthys flesus) has been examined by immunocytochemistry and in situ hybridization for the expression of parathyroid hormone-related protein (PTHrP) and calcium-sensing receptors (CaSR). The N-terminus nucleotide and deduced amino acid sequences of flounder PTHrP were determined and used to prepare oligonucleotide probes and homologous antiserum. The Dahlgren cells of the posterior spinal cord and their axons contained PTHrP protein which was also detected around the capillaries of the urophysis. PTHrP gene expression was abundant in the Dahlgren perikarya and axons in the spinal cord, but it was absent from nerve endings in the urophysis. Calcium-sensing receptor protein was present in the Dahlgren perikarya and axons, also with abundant gene expression, but there was neither protein nor mRNA in the urophysis. There were no apparent differences between freshwater- and seawater adapted fish in either CaSR or PTHrP expression in the caudal neurosecretory system. These observations suggest that Dahlgren cells produce PTHrP which may be released from axons abutting capillaries in the urophysis. However, the sensing of ionic calcium appears to be confined to the perikarya of the Dahlgren cells in the spinal cord neuropil, suggesting that they are responsive to calcium in the central nervous system rather than the general circulation. PMID- 12090396 TI - Composite cellular defence stratagem in the avian respiratory system: functional morphology of the free (surface) macrophages and specialized pulmonary epithelia. AB - Qualitative and quantitative attributes of the free respiratory macrophages (FRMs) of the lung--air sac systems of the domestic fowl (Gallus gallus variant domesticus) and the muscovy duck (Cairina moschata) were compared with those of the alveolar macrophages of the lung of the black rat (Rattus rattus). The birds had significantly fewer FRMs compared to the rat. In the birds, the FRMs were found both in the lungs and in the air sacs. Under similar experimental conditions, the most robust FRMs were those of the domestic fowl followed by those of the rat and the duck. Flux of macrophages onto the respiratory surface from the subepithelial compartment and probably also from the pulmonary vasculature was observed in the birds but not in the rat. In the duck and the domestic fowl, a phagocytic epithelium that constituted over 70% of the surface area of the blood-gas (tissue) barrier lines the atrial muscles, the atria and the infundibulae. The epithelial cells of the upper respiratory airways contain abundant lysosomes, suggesting a high lytic capacity. By inference, the various defence strategies in the avian lung may explain the dearth of FRMs on the respiratory surface. We counter-propose that rather than arising directly from paucity of FRMs, an aspect that has been over-stressed by most investigators, the purported high susceptibility of birds (particularly table birds) to respiratory ailments and afflictions may be explained by factors such as inadequate management and husbandry practices and severe genetic manipulation for fast growth and high productivity, manipulations that may have weakened cellular and immunological defences. PMID- 12090397 TI - Electrophysiological study of the rectosigmoid canal: evidence of a rectosigmoid sphincter. AB - Previous studies strongly suggest the presence of a sphincter at the rectosigmoid junction, an area with a mean length of 2.8 cm in adults, called the rectosigmoid canal (RSC). To find supporting evidence of a sphincteric function for the RSC, two recording electrodes were applied to each of the sigmoid colon (SC), RSC and rectum (R) in 11 subjects during operative repair of huge incisional hernias. The RSC, SC and R were individually stimulated by a further electrode and their pressures monitored by a three-channel microtip catheter. The variables of the slow waves or pacesetter potentials, recorded at rest from the RSC and R, were significantly higher than those of the SC. While the frequency and conduction velocity of pacesetter potentials of the RSC and R were similar, the potential pacesetter amplitude of the RSC was significantly higher. The increase of the electrical activity and pressure upon electrostimulation was significantly greater in the RSC than that of the SC or R. Electrostimulation led to an increase in pressure of all three areas, the RSC increase being significantly the greatest. The greater increase of the electrical activity and pressure of the rectosigmoid canal upon electrostimulation, compared to that of the SC or R, strongly supports the presence of a rectosigmoid sphincter. PMID- 12090398 TI - Bodyworlds. PMID- 12090399 TI - The function of the Periaxin gene during nerve repair in a model of CMT4F. AB - Mutations in the Periaxin (PRX) gene are known to cause autosomal recessive demyelinating Charcot-Marie-Tooth (CMT4F) and Dejerine-Sottas disease. The pathogenesis of these diseases is not fully understood. However, progress is being made by studying both the periaxin-null mouse, a mouse model of the disease, and the protein-protein interactions of periaxin. L-periaxin is a constituent of the dystroglycan-dystrophin-related protein-2 complex linking the Schwann cell cytoskeleton to the extracellular matrix. Although periaxin-null mice myelinate normally, they develop a demyelinating peripheral neuropathy later in life. This suggests that periaxin is required for the stable maintenance of a normal myelin sheath. We carried out sciatic nerve crushes in 6-week-old periaxin null mice, and, 6 weeks later, found that although the number of myelinated axons had returned to normal, the axon diameters remained smaller than in the contralateral uncrushed nerve. Not only do periaxin-null mice have more hyper myelinated axons than their wild-type counterparts but they also recapitulate this hypermyelination during regeneration. Therefore, periaxin-null mice can undergo peripheral nerve remyelination, but the regulation of peripheral myelin thickness is disrupted. PMID- 12090400 TI - Systematic reviews of treatment for inflammatory demyelinating neuropathy. AB - This review describes the progress made in preparing Cochrane systematic reviews of randomized controlled trials for Guillain-Barre syndrome (GBS), chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), multifocal motor neuropathy (MMN) and the demyelinating neuropathies associated with paraproteins. The discovery of antibodies against myelin and axolemmal glycolipids and proteins has not yet replaced the clinicopathological classification on which treatment trials have been based. Systematic reviews have endorsed the equivalence of plasma exchange (PE) and intravenous immunoglobulin (IVIg) and the lack of efficacy of steroids in GBS. Systematic reviews have also endorsed the value of steroids, PE and IVIg in CIDP but randomized controlled trials have only shown benefit from IVIg in MMN. There is a paucity of evidence concerning the efficacy of treatments in paraproteinaemic demyelinating neuropathy apartment from small trials showing short-term benefit from PE or IVIg. There is a lack of good quality controlled trials of immunosuppressive agents in any of these conditions. As the number of treatment trials increases, Cochrane systematic reviews will be an increasingly valuable resource for summarizing the evidence from randomised controlled trials on which to base clinical practice. They already demonstrate major deficiencies in the existing evidence base. PMID- 12090401 TI - Dejerine-Sottas syndrome grown to maturity: overview of genetic and morphological heterogeneity and follow-up of 25 patients. AB - Dejerine-Sottas syndrome (DSS) is an early onset demyelinating motor and sensory neuropathy with motor nerve conduction velocities below 12 m s(-1). The phenotype is genetically heterogeneous, and autosomal dominant (AD) as well as autosomal recessive (AR) inheritance is described. Nerve pathology is highly variable. It is generally presumed that clinical course is severe, leading to wheelchair dependency at an early age. In this study we documented the clinical and pathological features in 25 patients with a DSS and we evaluated the clinical course. In our series 14 patients had an AD mutation and six were probably affected by an AR disorder. In three patients inheritance mode was unknown and two patients obviously suffered from an acquired disorder. The clinical course in all patients was documented. Nine of the 25 patients showed a moderate handicap in adult life; walking distance was still at least 1 km. Age at last investigation of the ambulant patients ranged from 22 to 62 years (mean 38.6 years), and ambulant patients were found in all genetic subgroups. We conclude that DSS, although in general denoting a more serious neuropathy than CMT1, does not imply a severe disability or wheelchair dependency in adult life. PMID- 12090402 TI - Understanding Schwann cell-neurone interactions: the key to Charcot-Marie-Tooth disease? AB - Charcot-Marie-Tooth disease (CMT) comprises a heterogeneous group of disorders. The most frequent subtype is caused by increased PMP22 gene dosage or missense point mutations affecting the PMP22 gene (CMT type 1A; CMT1A). Animal models in rat and mouse with the corresponding PMP22 alterations are available and mimic many aspects of the human diseases. Detailed examinations of the animal mutants, together with complementary data from patients, point towards altered Schwann cell-neurone interactions as a major underlying mechanism of CMT1A and related hereditary neuropathies. This is evident from the finding that mutated proteins affecting either Schwann cells or neurones have a profound influence on their partner cells. Recently, a number of novel genes causing various forms of CMT have been identified which are expressed either mainly by Schwann cells and/or by the accompanying neurones. These genes can be viewed, in analogy to classic experiments routinely performed in lower vertebrates, as the result of a 'functional screen' revealing crucial players in the interactions between Schwann cells and neurones. Studying how Schwann cell and axon-encoded proteins are functionally interconnected will be an exciting task for the future. It will not only yield insights into the molecular and cellular basis of neuropathies but also provide crucial information about the interplay between Schwann cells and neurones in general. PMID- 12090403 TI - Signals that determine Schwann cell identity. AB - While the signals that direct neural crest cells to choose the glial lineage and generate Schwann cell precursors are still obscure, studies both in vivo and in vitro indicate that the survival and differentiation of these cells to form Schwann cells is regulated by at least two signals, neuregulin-1 and endothelin. We know little about the signals that cause some immature Schwann cells to choose myelin differentiation, while other cells form non-myelinating cells. Three transcription factors, Sox-10, Oct-6 and Krox-20, have been shown to play key roles in the Schwann cell lineage. The transcription factor Krox-20 has been identified as a major target of the signals that induce myelin differentiation. Gene transfer experiments in vitro show that this protein has a remarkable ability to promote a large number of phenotypic changes in immature Schwann cells that characterize the transition of these cells to myelinating cells. Furthermore, Krox-20 shows important functional interactions with neuregulin and transforming growth factor beta (TGFbeta), two factors that have been implicated in the regulation of myelination in postnatal nerves. Another signal of importance in developing peripheral nerves, Desert Hedgehog, secreted by Schwann cells directs formation of the peripheral nerve connective tissue sheaths. Ongoing gene screening experiments are likely to reveal new genes of interest in this system. PMID- 12090404 TI - Comparison of a new pmp22 transgenic mouse line with other mouse models and human patients with CMT1A. AB - Charcot-Marie-Tooth disease type 1A is a dominantly inherited demyelinating disorder of the peripheral nervous system. It is most frequently caused by overexpression of peripheral myelin protein 22 (PMP22), but is also caused by point mutations in the PMP22 gene. We describe a new transgenic mouse model (My41) carrying the mouse, rather than the human, pmp22 gene. The My41 strain has a severe phenotype consisting of unstable gait and weakness of the hind limbs that becomes obvious during the first 3 weeks of life. My41 mice have a shortened life span and breed poorly. Pathologically, My41 mice have a demyelinating peripheral neuropathy in which 75% of axons do not have a measurable amount of myelin. We compare the peripheral nerve pathology seen in My41 mice, which carry the mouse pmp22 gene, with previously described transgenic mice over-expressing the human PMP22 protein and Trembler-J (TrJ) mice which have a P16L substitution. We also look at the differences between CMT1A duplication patients, patients with the P16L mutation and their appropriate mouse models. PMID- 12090405 TI - De- and remyelination in spinal roots during normal perinatal development in the cat: a brief summary of structural observations and a conceptual hypothesis. AB - We have studied the perinatal development of large myelinated axons (adult D > 10 microm) in cat ventral and dorsal lumbosacral spinal roots using autoradiography and electron microscopy (serial section analysis). These axons acquire their first myelin sheaths 2-3 weeks before birth and show nearly mature functional properties first at a diameter of 4-5 microm, i.e. 3-4 weeks after birth. The most conspicuous event during this development takes place around birth, when a transient primary myelin sheath degeneration strikes already well myelinated although short 'aberrant' Schwann cells. The aberrant Schwann cells become completely demyelinated, then measuring about 10 microm in length, and are subsequently eliminated from their parent axons. Morphometry indicates that on average 50% of the Schwann cells originally present along a prospective large spinal root axon suffer elimination. Here it should be noted that in cat lumbo sacral spinal roots, the longitudinal growth of myelinated Schwann cells that belong to the group containing what will be the largest fibers is on average twice that of their parent axons. The elimination phenomenon is particularly striking in the dorsal roots close to the spinal cord where CNS tissue invades the root for several hundred micrometres. Our observations suggest that, once demyelinated and then eliminated, Schwann cells (i.e. aberrant Schwann cells) colonize neighbouring axons, future myelinated as well as future unmyelinated ones. In the former case the immigrant Schwann cells appear to start myelin production, possibly risking a second demyelination and elimination. We take our observations to indicate that Schwann cells in the cat, during normal development, may switch iteratively between a 'myelin-producing' and a 'non myelin-producing' phenotype. From a functional point of view the transient presence along a myelinated axon of intercalated unmyelinated segments approximately 10 microm long, due to aberrant Schwann cells, would mean a slowing down of the action potential. The rapid disappearance of aberrant Schwann cells during the two first postnatal weeks could then explain the progressing normalization of the leg-length conduction time. PMID- 12090406 TI - Role of immune cells in animal models for inherited neuropathies: facts and visions. AB - Mice heterozygously deficient in the peripheral myelin adhesion molecule P0 (P0+/ mice) are models for some forms of Charcot-Marie-Tooth (CMT) neuropathies. In addition to the characteristic hallmarks of demyelination, elevated numbers of CD8-positive T-lymphocytes and F4/80-positive macrophages are striking features in the nerves of these mice. These immune cells increase in number with age and progress of demyelination, suggesting that they might be functionally related to myelin damage. In order to investigate the pathogenetic role of lymphocytes, the myelin mutants were cross-bred with recombination activating gene 1 (RAG-1) deficient mice, which lack mature T- and B-lymphocytes. The immunodeficient myelin mutants showed a less severe myelin degeneration. The beneficial effect of lymphocyte-deficiency was reversible, since demyelination worsened in immunodeficient myelin-mutants when reconstituted with bone marrow from wild-type mice. Ultrastructural analysis revealed macrophages in close apposition to myelin and demyelinated axons. We therefore cross-bred the P0+/- mice with spontaneous osteopetrotic (op) mutants deficient in the macrophage colony-stimulating factor (M-CSF), hence displaying impaired macrophage activation. In the corresponding double mutants the numbers of macrophages were not elevated in the peripheral nerves, and the demyelinating phenotype was less severe than in the genuine P0+/- mice, demonstrating that macrophages are also functionally involved in the pathogenesis of genetically mediated demyelination. We also examined other models for inherited neuropathies for a possible involvement of immune cells. We chose mice deficient in the gap junction component connexin 32, a model for the X linked form of CMT. Similar to P0-deficient mice, T-lymphocytes and macrophages were elevated and macrophages showed a close apposition to degenerating myelin. We conclude that the involvement of T-lymphocytes and macrophages is a common pathogenetic feature in various forms of slowly progressive inherited neuropathies. PMID- 12090408 TI - Missing data: our view of the state of the art. AB - Statistical procedures for missing data have vastly improved, yet misconception and unsound practice still abound. The authors frame the missing-data problem, review methods, offer advice, and raise issues that remain unresolved. They clear up common misunderstandings regarding the missing at random (MAR) concept. They summarize the evidence against older procedures and, with few exceptions, discourage their use. They present, in both technical and practical language, 2 general approaches that come highly recommended: maximum likelihood (ML) and Bayesian multiple imputation (MI). Newer developments are discussed, including some for dealing with missing data that are not MAR. Although not yet in the mainstream, these procedures may eventually extend the ML and MI methods that currently represent the state of the art. PMID- 12090409 TI - Statistical power in randomized intervention studies with noncompliance. AB - This study examined various factors that affect statistical power in randomized intervention studies with noncompliance. On the basis of Monte Carlo simulations, this study demonstrates how statistical power changes depending on compliance rate, study design, outcome distributions, and covariate information. It also examines how these factors influence power in different methods of estimating intervention effects. Intent-to-treat analysis and complier average causal effect estimation are compared as 2 alternative ways of estimating intervention effects under noncompliance. The results of this investigation provide practical implications in designing and evaluating intervention studies taking into account noncompliance. PMID- 12090407 TI - Axons and glial interfaces: ultrastructural studies. AB - At most vertebrate nerve transitional zones (TZs) there is a glial barrier which is pierced by axons passing between the CNS and PNS. Myelinated axons traverse this in individual tunnels. The same is true of larger non-myelinated axons. This holds widely among the vertebrates, for example, the large motor axons of the sea lamprey Petromyzon (which also possess TZ specializations not found in mammals). Smaller non-myelinated axons traverse the TZ glial tunnels as fascicles and so the barriers are correspondingly less comprehensive for them. Accordingly, in nerves composed of non-myelinated axons, such as the vomeronasal or the olfactory, a TZ barrier stretching across the nerve is effectivelyabsent. The chordateAmphioxus differsfrom the vertebrates in lacking a TZ barrier throughout. Invertebrates also lack glial barriers at the TZs between ganglia and interconnecting nerve trunks. The glial barrier at the dorsal spinal root TZ (DRTZ) has considerable value for analysing protocols aimed at achieving CNS regeneration, because it provides a useful model of the gliotic reaction at sites of CNS injury. Also, it is especially amenable to morphometric analysis, and so enables objective quantification of different protocols. Being adjacent to the subarachnoid space, it is accessible for experimental intervention. The DRTZ was used to investigate the value of neurotrophin 3 (NT3) in promoting axon regeneration across the TZ barrier and into the CNS following dorsal root crush. It promoted extensive regeneration and vigorous non-myelinated axonal ensheathment. On average, around 40% of regenerating axons grew across the interface, compared with virtually none in its absence. These may have traversed the interface through loci occupied by axons prior to degeneration. Many regenerating axons became myelinated, both centrally and peripherally. PMID- 12090410 TI - Power in randomized group comparisons: the value of adding a single intermediate time point to a traditional pretest-posttest design. AB - Adding a pretest as a covariate to a randomized posttest-only design increases statistical power, as does the addition of intermediate time points to a randomized pretest-posttest design. Although typically 5 waves of data are required in this instance to produce meaningful gains in power, a 3-wave intensive design allows the evaluation of the straight-line growth model and may reduce the effect of missing data. The authors identify the statistically most powerful method of data analysis in the 3-wave intensive design. If straight-line growth is assumed, the pretest-posttest slope must assume fairly extreme values for the intermediate time point to increase power beyond the standard analysis of covariance on the posttest with the pretest as covariate, ignoring the intermediate time point. PMID- 12090411 TI - When constraints interact: a caution about reference variables, identification constraints, and scale dependencies in structural equation modeling. AB - In traditional approaches to structural equations modeling, variances of latent endogenous variables cannot be specified or constrained directly and, consequently, are not identified, unless certain precautions are taken. The usual method for achieving identification has been to fix one factor loading for each endogenous latent variable at unity. An alternative approach is to fix variances using newer constrained estimation algorithms. This article examines the philosophy behind such constraints and shows how their appropriate use is neither as straightforward nor as noncontroversial as portrayed in textbooks and computer manuals. The constraints on latent variable variances can interact with other model constraints to interfere with the testing of certain kinds of hypotheses and can yield incorrect standardized solutions with some popular software. PMID- 12090412 TI - A critique of the correlated trait-correlated method and correlated uniqueness models for multitrait-multimethod data. AB - The correlated trait-correlated method (CT-CM) and correlated uniqueness (CU) confirmatory factor analysis models for multitrait-multimethod data are critiqued. Although the CU model often returns convergent and admissible factor solutions when the CT-CM model does not, the CU model is shown to have theoretical and substantive shortcomings. On the basis of this critique, the authors recommend that the CT-CM model be regarded as the generally preferred model and that the CU model be invoked only when the CT-CM model fails. PMID- 12090413 TI - A comparison of maximum covariance and K-means cluster analysis in classifying cases into known taxon groups. AB - Maximum covariance (MAXCOV) is a method for determining whether a group of 3 or more indicators marks 1 continuous or 2 discrete latent distributions of individuals. Although the circumstances under which MAXCOV is effective in detecting latent taxa have been specified, its efficiency in classifying cases into groups has not been assessed, and few studies have compared its performance with that of cluster analysis. In the present Monte Carlo study, the classification efficiencies of MAXCOV and the k-means algorithm were compared across ranges of sample size, effect size, indicator number, taxon base rate, and within-groups covariance. When the impact of these parameters was minimized, k means classified more data points correctly than MAXCOV. However, when the effects of all parameters were increased concurrently, MAXCOV outperformed k means. PMID- 12090414 TI - Hierarchical modeling of sequential behavioral data: an empirical Bayesian approach. AB - The authors review the common methods for measuring strength of contingency between 2 behaviors in a behavioral sequence, the binomial z score and the adjusted cell residual, and point out a number of limitations of these approaches. They present a new approach using log odds ratios and empirical Bayes estimation in the context of hierarchical modeling, an approach not constrained by these limitations. A series of hierarchical models is presented to test the stationarity of behavioral sequences, the homogeneity of sequences across a sample of episodes, and whether covariates can account for variation in sequences across the sample. These models are applied to observational data taken from a study of the behavioral interactions of 254 couples to illustrate their use. PMID- 12090415 TI - Pediatric laboratory medicine: current challenges and future opportunities. AB - The practice of pediatric laboratory medicine involves unique challenges related to development, nutrition, growth, and diseases during different periods of infancy, childhood, and adolescence. This article discusses key aspects of pediatric laboratory medicine faced by clinical pathologists, clinical laboratory scientists, and clinicians, including point-of-care testing, preanalytic variables, analytic factors, age-specific reference intervals, esoteric laboratory tests, clinical impact, andfuture opportunities. Although challenging, pediatric laboratory testing offers many opportunities for improved patient care, clinical- and laboratory-based research, and education. PMID- 12090416 TI - Estimating the budgetary impact of setting the medicare clinical laboratory fee schedule at the national limitation amount. AB - The Institute of Medicine (IOM) of the National Academy of Sciences was commissioned by Congress to study the current system for the payment of diagnostic clinical laboratory services provided to Medicare beneficiaries. The current system was established in 1984 and has grown in complexity and is of diminishing contemporary relevance. The IOM recommended that a single, rational, nationalfee schedule be established and that it be initially based on the National Limitation Amount (NLA) currently mandated as the national fee cap. To estimate the potential budgetary impact of this recommendation, we merged the 1999 Part B Extract and Summary System and the 1999 Clinical Diagnostic Laboratory Fee Schedule (CLFS). By using an estimated 193 million allowed services from this data set and the current mean fee of $9.14 per test, current spending is approximately $1,768 million. The impact of raising the CLFS to the NLA will be approximately $1,792 million, or $9.26 per test. The estimated cumulative budgetary effect, factoring in the current forecast for the Consumer Price Index, is an increase of approximately $993 million over 5 years and $2,359 million over 10 years. PMID- 12090417 TI - Comparative study of marginal zone lymphoma involving bone marrow. AB - Few studies have characterized or compared the pathologic features of bone marrow involvement by extranodal (EMZL), splenic (SMZL), and nodal marginal zone lymphoma (NMZL). We evaluated 45 bone marrow biopsy specimens from 39 patients with marginal zone lymphomas. As previously reported, bone marrow involvement was frequent (100%) in patients with SMZL. We also identified lymphoma involving bone marrow in 11 (44%) of 25 patients with EMZL and 1 of 2 patients with NMZL. The patterns of infiltration were mixed in all groups; however, the extent of involvement was greater in SMZL than in EMZL. In addition, germinal centers were present in bone marrow biopsy specimens involved by lymphoma in 4 patients with SMZL. Intrasinusoidal infiltration was common (10/12 [83%]) and prominent in patients with bone marrow involvement by SMZL, but was not invariably present. Intrasinusoidal infiltration of the bone marrow also was not specific for SMZL since similar infiltrates, although subtle, also were found in patients with other small B-cell lymphoproliferative disorders, including 6 (55%) of 11 patients whose bone marrow samples were infiltrated by EMZL. PMID- 12090418 TI - Neutrophil-rich Ki-1-positive anaplastic large cell lymphoma: a study by fine needle aspiration biopsy. AB - Fine-needle aspiration biopsy (FNAB) is an accurate, cost-effective method of evaluating lymphomas. The neutrophil-rich variant of anaplastic large cell lymphoma (NR-ALCL) is a rare non-Hodgkin lymphoma. To our knowledge, we present thefirst study of NR-ALCL by FNAB cytology. Histologic confirmation was available for both patients. Both cases were positive for Ki-1 (CD-30) and were either T cell or null-cell phenotype. FNAB specimens were highly cellular with a single cell pattern composed of pleomorphic tumor cells, "hallmark" tumor cells, and a background rich in neutrophils that occasionally obscured tumor cells. Diagnosis on FNAB is difficult owing to the rarity of this tumor, its resemblance to Hodgkin lymphoma and other non-Hodgkin lymphomas that express CD30, its similarity to an infectious process, and its occasional confusion with metastatic carcinoma and melanoma. Reproducible cytologic features usually are present, and the diagnosis can be made conclusively by FNAB in conjunction with ancillary studies. PMID- 12090420 TI - Assessment of interlaboratory variation in the immunohistochemical determination of estrogen receptor status using a breast cancer tissue microarray. AB - The determination of tumor cell estrogen receptor (ER) expression status by immunohistochemical analysis has become standard practice, yet assay reproducibility has not been assessed adequately. By using a breast cancer tissue microarray, we examined interlaboratory variability in ER reporting. A 2-fold redundant tissue microarray block was created from 29 breast cancers. Unstained slides were distributed to 5 laboratories, and each laboratory immunostained and scored 1 slide for ER. Interlaboratory agreement ranged from moderate to high (overall kappa = 0.54 for 0-3+ grading; overall kappa = 0.84 for negative vs positive assessment of ER status). When 1 observer scored each of the 5 slides, interlaboratory agreement was slightly better (kappa = 0.63 for 0-3+ scoring; kappa = 0.96 for negative vs positive scoring). One laboratory, which had used an antibody and antigen retrieval method different from the others, demonstrated only fair concordance with the other 4 laboratories, but there was substantial intralaboratory interobserver agreement and excellent agreement with an outside observer reviewing the slide stained in that laboratory. The tissue microarray was an efficient and effective tool for identifying variability in ER reporting and should prove valuable in other external quality assurance programs. PMID- 12090419 TI - Acute lymphoblastic leukemia in elderly patients the Philadelphia chromosome may not be a significant adverse prognostic factor. AB - Acute lymphoblastic leukemia (ALL) in elderly patients (59 years or older) carries a poor prognosis, and this finding may be attributed to the relatively high frequency of the Philadelphia chromosome (Ph). To test this hypothesis, we reviewed the clinicopathologic features of 23 consecutive, newly diagnosed elderly patients with ALL (14 men, 9 women, aged 59-92 years) uniformly treated at our institution and compared the Ph+ and Ph- groups. Conventional cytogenetic data were available for 21 of 23 cases; 7 (33%) were Ph+. All Ph+ cases were of precursor B-cell type. The remaining 16 tumors were of precursor B-cell (10), mature B-cell (2), precursor T-cell (3), and mixed precursor T-cell/B-cell (1) type. Ph+ and Ph- groups did not differ significantly in median survival (13.4 months vs 19.0 months) or other variables studied. The Ph may not be a significant adverse prognostic factor in ALL in elderly patients. PMID- 12090421 TI - Cytokeratin immunohistochemical validation of the sentinel node hypothesis in patients with breast cancer. AB - No standard method for handling and histopathologic examination of the sentinel node (SN) exists. We hypothesized that a focused examination of all nodes with serial sectioning and cytokeratin immunohistochemical staining would confirm the SN as the node most likely to harbor metastasis. Intraoperative lymphatic mapping and sentinel lymphadenectomy using blue dye and (99m)technetium-labeled sulfur colloid were performed. All nodes were stained with H&E. All tumor-free nodes underwent additional sectioning and staining with H&E and an immunohistochemical stain. Routine H&E examination detected SN metastases in 27.6% of cases. Occult SN metastases were identified in 12.7% of cases. None of the 724 non-SNs examined contained occult metastases. The SN false-negative rate was zero. This study confirms histopathologically that the SN has biologic significance as the axillary node most likely to harbor metastatic tumor Standardization of the handling, sectioning, and staining of the SN is necessary as lymphatic mapping and sentinel lymphadenectomy become integrated into the care of patients with breast cancer PMID- 12090422 TI - Immunohistochemical localization of survivin in benign cervical mucosa, cervical dysplasia, and invasive squamous cell carcinoma. AB - Survivin is an inhibitor of apoptosis protein (IAP) that is expressed in fetal development and in cancer Survivin expression in premalignant lesions remains undefined. We obtained 73 samples of cervical squamous tissue, including 31 normal, 17 low- and 15 high-grade squamous intraepithelial lesions (LSILs, HSILs), and 10 squamous cell carcinomas (SCCs)from cone biopsy and hysterectomy specimens, and stained for survivin using an immunoperoxidase method. Nuclear staining was detected in normal mucosa, LSILs, and HSILs; staining intensity was greatest in cases with morphologic evidence of human papillomavirus (HPV) infection. In situ hybridization of serial sections demonstrated colocalization of HPV DNA and survivin. Cytoplasmic staining was observed in immature squamous metaplasia and in SCCs. Survivin expression in immature metaplastic squamous mucosa may reflect a rolefor survivin in normal squamous differentiation. However, the histologic correlation between nuclear staining and HPV infection suggests involvement of survivin in HPV-mediated disruption of normal cellular maturation. PMID- 12090423 TI - WT1, estrogen receptor, and progesterone receptor as markers for breast or ovarian primary sites in metastatic adenocarcinoma to body fluids. AB - In tissue sections, detection of the Wilms tumor susceptibility gene 1 (WTI) protein, the hormonal receptors for estrogen (ER) and progesterone (PR), and gross cystic disease fluid protein (GCDFP) are useful for diagnosing ovarian and breast adenocarcinomas. We evaluated these markers for cytology cell-block preparations from 96 effusion specimens (metastases from 29 breast, 22 ovarian, and 45 adenocarcinomas from other sites). WTI protein was reactive in 19 cases inetastatic from ovary (86%), 2 from breast (7%), and none from other sites (specificity; 97%). Of the metastatic breast carcinomas, 21(72%) were reactive for ER, 15(52%) for PR, and 13 (45%) for both (combined specificity, 84%). GCDEP was reactive in only 4 breast cancer cases (14%). Ovarian tumors also were frequently positive for ER (19 [86%]), PR (II [SO%]), or both (10 [45%]). WTI protein is an effective marker for ovarian adenocarcinoma, especially in ascites. The detection of ER and PR in metastatic adenocarcinoma from pleural or pericardial efflusions can distinguish breast from lung primary sites. Reactivity for ER and PR did not distinguish between breast and ovarian metastases; however; studies for WTI protein and GCDFP may aid in making this distinction. PMID- 12090424 TI - Quantifying the value of in-house consultation in surgical pathology. AB - In-house consultation is a well-known method to improve diagnostic accuracy and agreement, but the technique has not been well studied. We reviewed the results of in-house consultation in a large private hospital practice setting for a 1 month period and determined its effect on diagnostic accuracy using the final sign-out as the "gold standard." During this 1-month period, 352 cases were reviewed as in-house consultations. Initial complete agreement was found in 315 (89.5%) cases. Using the initial diagnosis as the test case and the final sign out as the gold standard, of the 37 discrepant cases, 4 (1.1%) were thought to represent false-negative results, (0.3%) a false-positive result, 3 (0.9%) differences in type, and 29 (8.2%) differences in diagnostic threshold. Disagreements in 10 cases were thought to be potentially clinically significant. Internal consultation was obtained on approximately 20% of all cases seen in the laboratory and disagreements were found in 2% of all cases. Internal consultation has a significant and measurable impact on the practice of surgical pathology. PMID- 12090425 TI - K-ras, p53, and DPC4 (MAD4) alterations in fine-needle aspirates of the pancreas: a molecular panel correlates with and supplements cytologic diagnosis. AB - Between January 1997 and February 2000, 101 fine-needle pancreatic aspirates were obtained. After a cytologic diagnosis was made, possible molecular alterations were determined on the 94 aspirates with adequate tissue using a molecular panel (K-ras, p53, and DPC4 [MAD4] genes). The 94 aspirates were categorized as follows: diagnostic of adenocarcinoma, 48 (51%); atypical (suggestive of but not diagnostic of adenocarcinoma), 19 (20%); negative for adenocarcinoma, 25 (2 7%); diagnostic of a neoplasm other than adenocarcinoma, 2 (2%). Clinical follow-up revealed that 3 patients (12%) with negative cytologic diagnoses and 12 patients (63%) with atypical cytologic diagnoses had adenocarcinoma. Of 63 with a final diagnosis of adenocarcinoma, 42 (67%) had an alteration in at least 1 of the genes analyzed. In contrast, only 2 (6%) of 31 patients without adenocarcinoma had an alteration in 1 gene on the panel. Overall, the molecular analyses supported the diagnosis of adenocarcinoma in 6 (32%) of 19 aspirates originally diagnosed as atypical by cytology alone. A molecular panel that includes the K ras, p53, and DPC4 (MAD4) genes correlates with and can supplement traditional cytologic diagnosis of pancreatic fine-needle aspirates. PMID- 12090426 TI - Lichenoid tissue reaction in malignant melanoma: a potential diagnostic pitfall. AB - Lichenoid tissue reactions can occur in malignant melanoma and may cause partial regression of the lesion. We studied a series of melanomas to determine how frequently lichenoid tissue reaction obscures the diagnosis of malignant melanoma. We retrospectively reviewed 342 cases of invasive malignant melanoma and melanoma in situ from the head, neck, chest, and back. Of the 342 cases, 23 (6.7%) had a lichenoid tissue reaction obscuring a portion of the lesion. In 6 cases (1.8%), the lichenoid tissue reaction replaced a major portion of the lesion. Knowledge of this phenomenon can prevent misdiagnosis. PMID- 12090427 TI - Uterine granulomas: clinical and pathologic features. AB - We retrospectively studied the clinical and pathologic features of uterine granulomas over a 10-year period. Granulomas were detected in 30 women, 22 to 81 years old, in the cervix (n = 12/1,090 cervical specimens; 1.1%) or uterine corpus (n = 18/12,000 uterine specimens; 0.15%). They were discovered during evaluation of abnormal bleeding, cytologic specimens, or other gynecologic conditions. None of the patients had constitutional symptoms. These granulomas were often focal (n = 25 [83%]), exhibiting features of foreign body-type (n = 17 [68%]); or they were diffuse (n = 5 [17%]), all with negative acid-fast bacilli or fungal stains and sometimes necrotizing (2 [40%]). Focal granulomas were highly associated with a preceding biopsy or surgery (22/25 vs 14/53 age-matched control subjects). Follow-up of 28 patients (median, 16 months) showed that 27 remained healthy; only 1 patient developed generalized lesions consistent with sarcoidosis 16 months later Uterine granulomas are rare. They are eitherfocal, related to previous biopsy or surgery, or diffuse, usually representing local reaction without an obvious cause. Association with infection or systemic granulomatous disorders is uncommon. PMID- 12090428 TI - Rasmussen encephalitis: a clinicopathologic and immunohistochemical study of seven patients. AB - We retrospectively reviewed the clinicopathologic features and immunohistochemical profiles of 7 patients with Rasmussen encephalitis (age range, 3.5-15 years at surgery). All had medically intractable seizures (6 months' to 7 years' duration); all but 1 developed unilateral hemiparesis. Histologically, all cases were characterized by leptomeningeal and parenchymal perivascular chronic inflammation consisting primarily of T lymphocytes (CD3+, CD5+, CD7+). In all but 1 case, a predominance of CD8+ T-cytotoxic/suppressor lymphoid cells over CD4+ cells was observed. All cases had rare B lymphocytes (CD79a+, CD20+). Rare CD10+ and no CD56+ cells were noted. All cases were marked by diffuse proliferation of microglial cells, highlighted on CD68 immunostaining. Focal microglial nodule formations were observed in 4 cases and focal cortical atrophy in 5 cases. Viral inclusions were not noted. There was no evidence of Epstein-Barr virus by LMP-1 antibody immunostaining. The histologic findings of Rasmussen encephalitis resemble those of viral meningoencephalitis. The pathologicfindings may be only focally present, and missed, if diagnosis is made or confirmed with biopsy alone. Most lymphoid cells have a T-cell immunophenotype, with a predominance of CD8+ cells in most cases. PMID- 12090429 TI - Telepathology for routine light microscopic and frozen section diagnosis. AB - Telepathology (TP) uses telecommunication linkages to electronically capture, store, retrieve, and transmit images to distant sites. We assessed the feasibility of a dynamic real-time TP system for light microscopic (LM) diagnosis of anatomic pathology specimens, including frozen sections. Six pathologists, in 2 separate periods, read a set of 160 retrospectively retrieved slides (80 of which were frozen sections) by TP and LM. Reading times were recorded. Diagnoses were compared with the reference diagnosis (established by a group of 5 independent pathologists) and graded on a scale of 0 to 2 (2, correct; 1, incorrect but no clinical impact; 0, incorrect with clinical impact). Overall, LM was more accurate than TP compared with the reference diagnosis (score, 1.68 vs 1.54). There was no difference in accuracy between frozen section and paraffin embedded tissue. Intraobserver agreement ranged from 82.5% to 88.2%. The average reading time was 6.0 minutes for TP and 1.4 minutes for LM. During the study, reading time decreased for TP but not for LM. These results show that despite marginally lower accuracy and longer reading times, TP isfeasible for routine light microscopic diagnosis, including frozen sections. PMID- 12090430 TI - MUC4 expression increases progressively in pancreatic intraepithelial neoplasia. AB - Pancreatic adenocarcinoma is believed to develop from histologically identifiable intraductal lesions known as pancreatic intraepithelial neoplasias (PanINs) that undergo a series of architectural, cytologic, and genetic changes, a progression model similar to the adenoma-carcinoma sequence in the colon. The apomucin MUC4 has been implicated in invasive pancreatic adenocarcinoma. MUC4 expression is not detectable at the RNA level in normal pancreas but is detectable at high levels in invasive pancreatic adenocarcinoma. We documented the pattern of expression of MUC4 in PanINs by studying a series of 71 PanIN lesions immunohistochemically using a new monoclonal antibody to MUC4. Five (17%) of 30 PanIN-1 lesions, 10 (36%) of 28 PanIN-2 lesions, 11 (85%) of 13 PanIN-3 lesions, and 25 (89%) of 28 invasive adenocarcinomas labeled with the MUC4 antibody used in the study. In addition, afew nonneoplastic lesions labeled with the MUC4 antibody, including reactive ducts in chronic pancreatitis, atrophic ducts filled with inspissated secretions, and ducts showing squamous metaplasia. Our data help establish the patterns of MUC4 expression in neoplastic precursors in the pancreas and add further support to the progression model for pancreatic adenocarcinoma. PMID- 12090431 TI - Lobular neoplasia in breast core needle biopsy specimens is not associated with an increased risk of ductal carcinoma in situ or invasive carcinoma. AB - Recent reports suggest that the finding of lobular neoplasia (atypical lobular hyperplasia [ALH] or bular carcinoma in situ [LCIS]) in breast core needle biopsy specimens may be associated with an increased risk of both ductal carcinoma in situ (DCIS) or invasive carcinoma at excision. We reviewed our breast core biopsy material to see if we could confirm this finding. from 4,297 biopsies, 71 cases of lobular neoplasia lone and 35 cases of lobular neoplasia associated with typical ductal hyperplasia were identified. Biopsy follow-up revealed DCIS or invasive carcinoma in none of 6 cases of ALH, none of 9 cases of LCIS, and DCIS in 1 of 11 cases with both atypical ductal hyperplasia and LCIS. Our results suggest that patients with lobular eoplasia in breast core biopsy specimens are not at increased risk of either DCIS or invasive carcinoma at excision, and patients with this finding and no other linical or pathologic indications for biopsy can be llowed up rather than routinely undergo excision. PMID- 12090432 TI - Diurnal variation of serum iron, iron-binding capacity, transferrin saturation, and ferritin levels. AB - Serum iron levels vary throughout the day. Morning levels are generally assumed to be higher than afternoon or evening levels. We studied whether our practice of restricting serum iron collections to the morning was necessary. Serum iron, iron binding capacity, transferrin saturation, and ferritin levels were determined on blood specimens obtained from 20 healthy adult volunteers at 8 AM, noon, and 4 PM (day 1) and 8 AM (day 2). Although statistically significant differences among mean values for the collection times were observed for iron, iron-binding capacity, and (log) ferritin, no consistent diurnal variation was seen. Morning iron levels were higher than afternoon levels for only half of the subjects. Between-day variation for all 4 analytes was similar to within-day variation. We conclude that the practice of restricting iron specimen collections to a specific time of day does not improve the reliability of the test result. PMID- 12090433 TI - Cost analysis of a neonatal point-of-care monitor. AB - A hypothetical model using a base case and sensitivity analyses compared averted and incurred costs of in-line monitoring with neonatal intensive care unit satellite laboratory testing. Data were obtained retrospectively for 1 year from 50 consecutive critically ill premature neonates weighing less than 1,000 g at birth whose blood tests were performed in-line and processed at the satellite laboratory. Averted costs included phlebotomies, satellite blood testing, and transfusions; incurred costs included in-line monitor rental, nursing time, and daily monitor validation. In-line monitoring led to cost savings of $324 per neonate and a benefit/cost ratio (BCR) of 1.23 in our base case. Sensitivity and scenario analyses addressed uncertainty and led to a BCR variation of 0.41 to 2.48. Compared with satellite laboratory testing, in-line monitoring of critically ill neonates may generate cost savings through reduced laboratory analysis expense, less phlebotomy loss, and fewer blood transfusions for hospitals with high laboratory cost structures. Because most cost savings result from offsetting indirect costs (eg, building space and hospital overhead) that are of a longer term nature, short-run cost savings are less likely to be realized. PMID- 12090434 TI - Transient stress lymphocytosis: an immunophenotypic characterization of the most common cause of newly identified adult lymphocytosis in a tertiary hospital. AB - We prospectively evaluated 52 consecutive cases of newly identified absolute lymphocytosis to determine the hematologic and immunophenotypic features of transient stress lymphocytosis (TSL). The lymphocytosis in all cases was associated with an acute stressful event and ranged from 4,000 to 10,400/microL (4.0-10.4 x 10(9)/L). Compared with healthy individuals, patients with TSL showed an increase in the total WBC, absolute lymphocyte (ALC), absolute neutrophil (ANC), and platelet counts with no difference in hemoglobin levels. Immunophenotypic analyses of 38 cases revealed increases in absolute numbers of T B, and natural killer cells. Both CD4+ and CD8+ T cells were increased, predominantly accountedfor by an increase in memory cell subsets, with no change in gamma/delta T cells. Follow-up studies showed a significant reduction in the ALC with a concurrent increase in the ANC and reduction in hemoglobin values. The reduction in lymphocytes at resolution was accompanied by reduction in all broad Iymphocyte subsets. However, naive and memory subsets showed different patterns of alteration within the CD4+ and CD8+ populations, suggesting that acute stress differentially affects the in vivo distribution of these subsets. PMID- 12090435 TI - Hipe is better than hype for Irish healthcare. PMID- 12090436 TI - Acute stroke care: are we getting the basics right? PMID- 12090437 TI - Vascular surgery of the upper limb: the first year of a new vascular service. AB - Upper limb vascular reconstruction represents a small part of the vascular surgical workload (5%). The aim of this study was to assess the incidence of upper limb vascular reconstruction in a Regional Hospital. During the first year of a new vascular surgical service in Waterford Regional Hospital, upper limb vascular problems were prospectively analysed. Upper limb vascular reconstruction comprised seven of the total 92 major vascular procedures performed. Three cases were emergencies and four elective. There was one case of penetrating injury, two injuries due to blunt trauma, three patients with thoracic outlet syndrome (TOS) and one chronic ischaemia. The male:female ratio was 4:3 and the mean age was 42 years. Six of the cases were arterial in nature only, and one was both venous and arterial. Two of the cases were associated with upper limb fractures and multiple trauma. Three patients had interposition reversed cephalic vein grafting. One patient had an embolectomy and endarterectomy. Procedures for TOS included excision of a cervical rib in two patients (one bilateral) and scalenectomy alone in one patient. Of these, one patient also had thrombolysis and thrombectomy of the axillary and brachial artery. All of the patients made a good functional recovery and all arteries remained patent but the patient with the brachial plexus injury is awaiting repair abroad. Upper limb vascular problems form a small but significant part (8%) of the workload. Many cases present as emergencies and maybe associated with multiple trauma. This emphasises the need for an emergency vascular surgery service in all trauma units. PMID- 12090438 TI - Elective total hip arthroplasty in patients aged 85 years and older. AB - Previous reports indicate that elderly patients suffer more complications than younger patients undergoing total hip arthroplasty (THR). We reviewed 60 consecutive elderly patients undergoing THR. Statistical comparisons were made with a control group of patients. The average age was 86.6 (range 85-92) years. Pre-operative D'Aubigne-Postel Score averaged 8.4 (range 1-14) points, post operative D'Aubigne-Postel Score averaged 13.1 (range 9-18) points. Compared to the control group, elderly patients had an increased intra-operative blood loss, increased blood transfusion rates, (p=0.0005) and a longer hospital stay (p = 0.0002). Comparing D'Aubigne-Postel scores, there was no significant difference between the two groups (p<0.001). We conclude that THR in elderly patients is a safe procedure and leads to functional improvements similar to those seen in younger counterparts. PMID- 12090439 TI - Severe acute alcoholic hepatitis: an audit of medical treatment. AB - Despite advances in treatment, severe alcoholic hepatitis is still associated with a high mortality rate of 30% to 40%. Nutritional support and steroids in selected patients are believed to improve prognosis. In controlled trials steroids have been beneficial in patients with a discriminant function (DF) value >32 or spontaneous hepatic encephalopathy. The aim of this study was to investigate current practice and outcomes in the treatment of acute alcoholic hepatitis. We retrospectively studied patients admitted to our unit with acute alcoholic hepatitis over a 4 year period. Forty-three patients with acute alcoholic hepatitis were admitted between 1994 and 1997. Overall mortality was 26% (11/43). Only 5 patients were treated with steroids of whom 1 died (mortality 20%). Liver biopsy was available in 19/43 of whom 12/19 (63%) had underlying cirrhosis in addition to alcoholic hepatitis. Mortality was higher in patients with a discriminant function of greater than 32 but not significantly so (32%: 8/25 vs 17%: 3/18 p = 0.31). A discriminant function of greater than 32 and contra-indications to steroid use was the best predictor of mortality (60% 6/10 P = 0.0096) compared to patients not fulfilling these criteria In this study overall mortality was comparable with published reports. Of interest was the relatively low liver biopsy rate and the fact that steroids were used in only a minority of eligible patients. We found that mortality was concentrated in a subgroup of patients with a discriminant function value >32 and contra indications to steroids. These criteria appear to identify a high-risk subgroup of patients. If confirmed, experimental treatments need to be targeted at this group to improve the overall prognosis of acute alcoholic hepatitis. PMID- 12090440 TI - Smoking: the major risk factor for SIDS in Irish infants. PMID- 12090441 TI - Dramatic response to nebulized morphine in an asthmatic patient with severe chronic cough. AB - We report a 56 year old man with a 16 year history of bronchial asthma who suffered severe coughing spasms that prevented employment, greatly reduced exercise tolerance and caused profound social embarrassment. His cough failed to respond to all standard asthma therapy and nebulized lignocaine. The patient was commenced on nebulized morphine sulphate with dramatic reduction in cough and improved quality of life. This benefit has been maintained at 2.5 years of follow up. PMID- 12090442 TI - National incidence of type 1 diabetes in childhood and adolescence. AB - The Republic of Ireland is considered a region of low type 1 diabetes incidence compared with the British Isles and the rest of Europe. To establish a baseline national incidence of type 1 diabetes, in those under 15 years, with a measure of case ascertainment. Prospective active monthly reporting of incident cases by paediatricians was undertaken, with survey of adult physicians, endocrinologists and nurse specialists nationally. A secondary source of case ascertainment was employed. The crude incidence rate of type 1 diabetes was 16.6 per 100,000 per year and the directly standardised incidence rate was 16.3 per 100,000 per year. The Republic has a high incidence of type 1 diabetes. Services should be planned and resources allocated accordingly. A register should be established to monitor changes in this important disease. Further study is required to explore differing incidence in Northern and Southern Ireland. PMID- 12090443 TI - Prevalence of obesity, lipid and glucose abnormalities in outpatients prescribed clozapine. AB - Individuals with schizophrenia have standardised mortality rates which are double that of the general population. In addition to suicide, high rates of cardiovascular and respiratory disease contribute to this raised mortality rate. Although clozapine has been reported to improve psychotic symptoms and decrease suicide rates, attention has recently focussed on its potential to increase cardiovascular risk factors including obesity, dyslipidemia and diabetes mellitus. This study aimed to ascertain the prevalence of these risk factors in a cohort of Irish outpatients treated with clozapine. PMID- 12090444 TI - A lump on the tongue: a diagnostic dilemma? AB - This paper is a case report of an oral lesion detected in a female patient in her twenties. The aetiology and pathology was not immediately clear-cut but the patient's history provided a clue to its origins and histopathological examination confirmed the diagnosis. It was a "Pregnancy Tumour" which had failed to regress after parturition. This case report will make medical and dental practitioners more aware of lumps and lesions in the mouth. It will also provide a more rational and scientific approach to the management of "Pregnancy Tumours". PMID- 12090445 TI - Neonatal seizures-management and outcome in a regional unit. PMID- 12090446 TI - Hemiballismus. PMID- 12090448 TI - Introduction to the proceedings of the Fifth International Conference on Breath Odour Research. AB - These proceedings represent a sample of the many fine papers that were presented at the Fifth International Conference on Breath Odour Research held in Tokyo on 2 3 July 2001. Space limitations precluded publication of all the papers, and the ones selected illustrate the diversity of approaches and the international nature of breath odour research. PMID- 12090449 TI - Classification and examination of halitosis. AB - In this paper, the classification of halitosis and the examination procedures used in diagnosing halitosis are outlined. Halitosis is classified into categories of genuine halitosis, pseudo-halitosis and halitophobia. Genuine halitosis is subclassified into physiologic halitosis and pathologic halitosis. Pathologic halitosis itself is subdivided into oral and extraoral halitosis. Patients diagnosed with pseudo-halitosis and halitophobia usually complain about having oral malodour that does not really exist. Pseudohalitosis can be treated by dental practitioners, but halitophobic patients must be referred to psychological specialists. Oral malodour can be measured using an organoleptic measurement or a gas chromatography analysis. The organoleptic measurement is the most practical procedure with which one can evaluate oral malodour. Gas chromatography (GC) analysis using a flame photometric detector has been shown to be the gold standard for measuring oral malodour, owing its reputation to its objectivity and reproducibility. Moreover, GC is specific for volatile sulphur compounds (VSC), which are the main causes of oral malodour. It has been demonstrated that there is a high correlation between the intensity of oral malodour and the VSC concentration as measured by GC. PMID- 12090450 TI - Treatment needs (TN) and practical remedies for halitosis. AB - Dental practitioners have traditionally neglected halitosis despite its high priority for the public, but practitioners' interest in halitosis has recently increased. Although oral pathologic or physiologic halitosis is easily reduced by a suitable treatment based on the treatment needs, systemic and psychological conditions sometimes confuse practitioners. Since a halitophobic patient never agrees with the result that his/her oral malodour has been reduced or eliminated after treatment, this may cause a dilemma for practitioners. Generally, halitosis patients, even genuine ones, have different psychological characteristics concerning their own breath than other individuals. Adverse psychological aspects of these patients are often promoted by the practitioner's mismanagement. Treatment Needs (TN) were, therefore, established to prevent practitioners' mismanagement of halitosis patients. By following these TN, patients can receive proper treatments for halitosis. However, to choose proper treatment measures, practitioners must refer to articles published in peer-reviewed journals, then use critical thinking to judge whether a product is effective in reducing oral malodour. Although it is challenging for dental practitioners to deal with patients with psychological conditions such as pseudo-halitosis or halitophobia, if appropriate treatments are administered accurately the practitioner does not risk mismanagement. PMID- 12090451 TI - Tongue brushing and mouth rinsing as basic treatment measures for halitosis. AB - Tongue brushing and mouth rinsing are basic treatment measures for halitosis, and as such are categorised as treatment needs (TN)-1. Although TN-1 is used for treatment of physiologic halitosis treatment, pseudo-, extra oral pathologic or halitophobic patients must also be managed with TN-1 as well as other treatments. Since the origin of physiological halitosis is mainly the dorso-posterior region of the tongue, tongue cleaning is more effective than mouth rinsing. However, practitioners should always instruct their patients on how to brush their tongues to prevent harmful effects. Another approach using a chlorhexidine mouthwash is most effective in reducing oral malodour. However, chlorhexidine should not be used routinely; therefore, zinc-containing mouthwashes have been recommended for use. People can also use chewing gum to reduce oral malodour. Surprisingly, however, it has been noted that sugarless chewing gum increased methyl mercaptan, one of the principal components of oral malodour. Mint did not reduce the concentration of methyl mercaptan either, although these products are widely used for their ability to mask oral malodour. There is a need for the development of a novel food or chewing gum that could considerably reduce VSC levels in mouth air to complement TN-1. PMID- 12090452 TI - Halitosis prevention campaign: a report of oral health promotion activities in Japan. AB - A local dental association, Hakodate Dental Association, has been conducting an oral health promotion campaign, 'Good-bye Bad Breath', in a local community since April 1998. The purpose of this campaign was not only to prevent oral malodour but also to raise people's awareness about oral health. Oral health education has been provided at schools and through television commercials, radio programmes and newspaper articles. Campaign posters and leaflets on bad breath have also been distributed to drug stores, medical clinics and schools, as well as dental clinics. It has been emphasised that oral malodour is a preventable condition and that the dentist can provide adequate advice and suitable treatment. Medical and pharmaceutical associations have cooperated with the Dental Association in running the campaign. Dental care was provided free of charge to the local residents. The free services provided included the assessment of oral malodour using a portable sulphide detector, instructional programmes, and consultations with dentists. This campaign offered a good opportunity to encourage people to visit the dentist for regular check-ups. The number of regular check-ups and halitosis patients has increased in 70% of the clinics belonging to the Dental Association. PMID- 12090453 TI - Halitosis in medicine: a review. AB - This review deals with the different forms of halitosis. Halitosis can be subdivided according to its original location. At present, halitosis of oral origin is quite well understood and some excellent reviews have already appeared in the literature. Special attention is given here to extra-oral halitosis. Extra oral halitosis can be subdivided into: halitosis from the upper respiratory tract including the nose; halitosis from the lower respiratory tract; blood-borne halitosis. In blood-borne halitosis, malodourant compounds in the bloodstream are carried to the lungs where they volatilise and enter the breath. Potential sources of blood-borne halitosis are some systemic diseases, metabolic disorders, medication and certain foods. The methods of analysis of halitosis are critically reviewed. Attention is also given to odour characterisation of various odourants. PMID- 12090454 TI - Gastrointestinal diseases and halitosis: association of gastric Helicobacter pylori infection. AB - The relationship between gastrointestinal conditions and halitosis is discussed. Few reports have suggested that gastrointestinal diseases may cause halitosis. H. pylori infection, which causes gastric ulcers, is considered as a possible cause for halitosis. Intensity of malodour of mouth air was found to be higher in H. pylori-positive patients than in negative patients. The levels of hydrogen sulphide and dimethyl sulphide in mouth air were also significantly higher in the positive patients than in the negative patients (P<0.05). When odour strength in exhaled breath was compared between the two groups, no significant difference was found. Hence, H. pylori infection might not cause a systemic condition producing breath odour. Although there were no significant differences in periodontal parameters or tongue coating between the positive and negative groups, H. pylori may be a frequent contributor to the production of malodour even though its role had not been suspected before. Further study would be necessary to clarify the reason for the increase of volatile sulphur compounds (VSCs) level in H. pylori infection. PMID- 12090456 TI - Methyl mercaptan production by periodontal bacteria. AB - Oral malodour is principally caused by volatile sulphur compounds (VSC) such as hydrogen sulphide, methyl mercaptan and dimethyl sulphide. Methyl mercaptan is highly toxic, and its presence within a periodontal pocket suggests involvement in the induction and/or progression of periodontal disease. Methyl mercaptan is produced from L-methionine by L-methionine- alpha -deamino- gamma mercaptomethane-lyase (METase). METase catalyses the alpha,gamma-eliminating reaction of L-methionine, which results in the release of alpha-ketobutyrate, methyl mercaptan and ammonia. Although methyl mercaptan is produced by a variety of microorganisms, Porphyromonas gingivalis is considered to be the most potent producer. METases of P. gingivalis have been characterised and the genes responsible for their production, the mg/genes, have been sequenced. To ascertain the role of METase in P. gingivalis pathogenicity, a METase-deficient mutant strain (M1217) from P. gingivalis strain W83 was engineered. Only 7.7% of the mice infected with W83 survived 4 days after subcutaneous injection, whereas 36% of the mice infected with M1217 survived over the same time period. Many papers have reported the periodontal pathogenesis of VSC. It has been argued that methyl mercaptan may play a significant role in the pathogenicity of P. gingivalis. PMID- 12090455 TI - The relationship between the presence of periodontopathogenic bacteria in saliva and halitosis. AB - OBJECTIVE: To evaluate the association between the presence of periodontal pathogenic bacteria in saliva and halitosis in mouth air. DESIGN: Cross-sectional microbiological and clinical oral examination of adult patients. SUBJECTS: 101 adult patients (25 males, 76 females) who attended the Preventive Dentistry and Breath Odour Clinic of Kyushu Dental College. Their average age was 50.0+/-13.5 years old (mean +/- SD). SETTING: The subjects were classified into three groups: halitosis subjects with a probing depth (PD) > or = 4mm (P group), halitosis subjects without PD > or = 4mm (H group), and non-halitosis subjects without PD > or = 4mm (C group). METHODS: All subjects received a periodontal examination. Volatile sulphur compounds (VSC: hydrogen sulphide and methyl mercaptan) were measured using gas chromatography. The presence of Bacteroides forsythus, Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Prevotella intermedia in the saliva was detected by polymerase chain reaction. RESULTS AND CONCLUSION: The presence of B. forsythus, P. gingivalis and P. intermedia influenced the production of VSC. Specifically, the presence of B. forsythus in subjects with periodontitis was strongly correlated to the concentration of VSC in mouth air. PMID- 12090457 TI - Cysteine challenge testing: a powerful tool for examining oral malodour processes and treatments in vivo. AB - Gram-negative oral bacteria rapidly produce the odorant hydrogen sulphide from cysteine. It provides a major part of the oral malodour bouquet while causing a corresponding decrease in the oxidation-reduction potential (Eh). A low Eh favours oral putrefaction and malodour occurrence. Challenge testing with cysteine (5ml of 6mM for 30 seconds) enabled evaluation of: the contribution of tongue and teeth bacteria to overall oral malodour; the effectiveness of tongue and tooth brushing, tooth scraping, gum chewing and mouthrinsing with different agents in reducing oral malodour. Successive cysteine challenge tests for 20 minute periods at selected times in a seven hour experiment were effective for assessing the magnitude and duration of an agent's effectiveness. Brushing the teeth reduced malodour modestly. So did tongue scraping and gum chewing. In contrast, brushing the tongue dorsum, especially the posterior half was remarkably effective, which confirmed it as a major site of oral malodour contribution. Rinses containing various actives showed wide variation in effectiveness. The experiments demonstrated that cysteine challenge testing is potentially a aluable tool for assessing the ability of the oral bacteria to produce malodour and for assessing agents designed to inhibit such production. PMID- 12090458 TI - Effect of deglycosylation of salivary glycoproteins on oral malodour production. AB - Putrefaction of saliva is commonly used as an in-vitro assay in oral malodour investigations. AIM: To exam the hypothesis that deglycosylation of salivary glycoproteins promotes oral malodour production. DESIGN: Porphyromonas gingivalis mediated putrefaction of salivary glycoproteins was tested following preincubation of saliva in the presence of beta-galactosidase with or without glycosidic inhibitor (galactosamine), and in the presence of glucose with or without a non-glycosylated protein (bovine serum albumin). METHODS: Malodour was determined by two odour judges, and volatile sulphides by using a sulphide monitor. Salivary glycoprotein degradation was measured densitometrically following electrophoresis on SDS-PAGE. RESULTS: The addition of beta galactosidase promoted salivary glycoprotein degradation and concomitant malodour production, whereas addition of a glycosidic inhibitor (D-galactosamine) inhibited this process. Glucose inhibited salivary glycoproteins putrefaction, but this inhibitory effect was mitigated when a non-glycosylated protein (BSA) was added. CONCLUSIONS: Deglycosylation of salivary glycoproteins may be an initial step in oral malodour production. This process exposes the protein core of the glycoprotein, which is then further degraded by Gram-negative microorganisms under anaerobic conditions. PMID- 12090459 TI - The significance of the source of zinc and its anti-VSC effect. AB - The anti-VSC (volatile sulphur compounds) effect of zinc is known to be associated with free zinc ions. OBJECTIVE: To examine whether zinc salts with low stability constants were more suitable as sources of zinc in zinc lozenges than zinc salts with high stability constants. The former provide free zinc ions upon dissolution in water, whereas the latter provide few such ions. DESIGN AND PARTICIPANTS: Identical lozenges were produced which contained either zinc acetate, zinc gluconate (low stability constants), zinc citrate or amino-acid chelated zinc (extremely high stability constants). All the lozenges contained 0.1 per cent of zinc. A test panel of 10 volunteers used the different lozenges randomly. VSC were measured by GC. RESULTS AND CONCLUSION: The lozenge with the highest stability constant was as effective as those with very low stability constants. The anti-VSC effect was thus not related to this constant. These findings may be explained by the possibility that alternative ligands with stronger affinity for zinc than the original ligands in the lozenges may be present in the oral cavity. An in vitro experiment indicated that the sulphide ion (S2-) may be such a ligand. PMID- 12090460 TI - Role of saliva in oral dryness, oral feel and oral malodour. AB - Patients who believe they have oral malodour often have a dry mouth condition instead. Here we have examined its relation to oral malodour, real or perceived. A direct relationship between the thickness of the film of residual saliva on mucosal surfaces throughout the mouth and perception of a dry mouth was observed. On the hard palate, the thickness of this film proved to be diagnostic for a dry mouth and corresponded to lower resting saliva flow and pH levels (P< 0.001). Intra-muscular administration of the anti-sialogogue, Robinul, accurately produced the dry mouth condition. Using a sulphide monitor, loss of volatile sulphur compounds into mouth air progressively occurred as the mouth became drier. Mouth pH and Eh on the dorsum of the tongue correspondingly fell. Mouth breathing led to tongue and palate moisture loss thus enabling escape of malodour volatiles into mouth air. Measurement of oral dryness should make it possible to differentiate genuine malodour from dry mouth related pseudo-malodour subjects, and in turn, the latter from patients that are halitophobic. This should facilitate identification of such patients and avoid error in their clinical management. PMID- 12090461 TI - The effects of methyl mercaptan on epithelial cell growth and proliferation. AB - AIM: Previous studies have demonstrated that methyl mercaptan (CH3SH), one of the main causes of oral malodour, might contribute to the initiation and progression of periodontal disease. These studies suggested that CH3SH may affect the epithelial cells of the gingival crevice, which form a barrier to the penetration of microbial substances. In this study, the effects of CH3SH on the epithelial cells and gingival fibroblasts were investigated. METHOD: Human oral epithelial carcinoma cell line (KB), human oral squamous cell carcinoma cell line (HSC-2), and human gingival fibroblasts (HGF) derived from healthy gingiva were used in this study. These cells were cultured in conditions of 5% CO2/95% air with or without CH3SH (10 ng/ml or 50 ng/ml) for 5 days. Cell numbers, proliferation and cytotoxicity were evaluated. RESULTS: CH3SH inhibited epithelial cell growth and proliferation at the concentration of 50 ng/ml, and a cytotoxic effect of CH3SH was also noted. On the other hand, HGF cells were not affected by 50 ng/ml CH3SH. CONCLUSION: High concentrations of CH3SH such as 50 ng/ml have an inhibitory effect on the growth and proliferation of epithelial cells, but not on those of fibroblasts. PMID- 12090462 TI - Chemotactic signaling pathways in neutrophils: from receptor to actin assembly. AB - In this review, we present an overview of the signaling elements between neutrophil chemotactic receptors and the actin cytoskeleton that drives cell motility. From receptor-ligand interactions, activation of heterotrimeric G proteins, their downstream effectors PLC and PI-3 kinase, the activation of small GTPases of the Rho family, and their regulation of particular cytoskeletal regulatory proteins, we describe pathways specific to the chemotaxing neutrophil and elements documented to be important for neutrophil function. PMID- 12090463 TI - Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) as modulators of both innate and adaptive immunity. AB - The structurally related neuropeptides VIP and PACAP are released within the lymphoid organs following antigenic stimulation, and modulate the function of inflammatory cells through specific receptors. In activated macrophages, VIP and PACAP inhibit the production of pro-inflammatory agents (cytokines, chemokines, and nitric oxide), and stimulate the production of the anti-inflammatory cytokine IL-10. These events are mediated through the VIP/PACAP effects on de novo expression or nuclear translocation of several transcription factors, i.e., NFkappaB, CREB, c-Jun, JunB, and IRF-1. The in vivo administration of VIP/PACAP results in a similar pattern of cytokine and chemokine modulation, which presumably mediates the protective effect of VIP/PACAP in septic shock. In addition, VIP/PACAP reduce the expression of the co-stimulatory molecules B7.1/B7.2, and the subsequent stimulatory activity of macrophages for T-helper cells. In T-cells expressing specific VIP/PACAP receptors, VIP and PACAP inhibit the expression of FasL through effects on NFkappaB, NFAT, and Egr2/3. The reduction of FasL expression has several biological consequences: inhibition of antigen-induced cell death in CD4 T-cells, inhibition of the FasL-mediated cytotoxicity of CD8 and CD4 effectors against direct and bystander targets, and promotion of long-term memory Th2 cells, through a positive effect on the survival of Th2, but not Th1, effectors. The various biological effects of VIP and PACAP are discussed within the range of a general anti-inflammatory model. PMID- 12090464 TI - Cysteine peptidases of mammals: their biological roles and potential effects in the oral cavity and other tissues in health and disease. AB - Cysteine peptidases (CPs) are phylogenetically ubiquitous enzymes that can be classified into clans of evolutionarily independent proteins based on the structural organization of the active site. In mammals, two of the major clans represented in the genome are: the CA clan, whose members share a structure and evolutionary history with papain; and the CD clan, which includes the legumains and caspases. This review focuses on the properties of these enzymes, with an emphasis on their potential roles in the oral cavity. The human genome encodes at least (but possibly no more than) 11 distinct enzymes, called cathepsins, that are members of the papain family C1A. Ten of these are present in rodents, which also carry additional genes encoding other cathepsins and cathepsin-like proteins. Human cathepsins are best known from the ubiquitously expressed lysosomal cathepsins B, H, and L, and dipeptidyl peptidase I (DPP I), which until recently were considered to mediate primarily "housekeeping" functions in the cell. However, mutations in DPP I have now been shown to underlie Papillon Lefevre syndrome and pre-pubertal periodontitis. Other cathepsins are involved in tissue-specific functions such as bone remodeling, but relatively little is known about the functions of several recently discovered enzymes. Collectively, CPs participate in multiple host systems that are active in health and in disease. They are involved in tissue remodeling and turnover of the extracellular matrix, immune system function, and modulation and alteration of cell function. Intracellularly, CPs function in diverse processes including normal protein turnover, antigen and proprotein processing, and apoptosis. Extracellularly, they can contribute directly to the degradation of foreign proteins and the extracellular matrix. However, CPs can also participate in proteolytic cascades that amplify the degradative capacity, potentially leading to pathological damage, and facilitating the penetration of tissues by cancer cells. We know relatively little regarding the role of human CPs in the oral cavity in health or disease. Most studies to date have focused on the potential use of the lysosomal enzymes as markers for periodontal disease activity. Human saliva contains high levels of cystatins, which are potent CP inhibitors. Although these proteins are presumed to serve a protective function, their in vivo targets are unknown, and it remains to be discovered whether they serve to control any human CP activity. PMID- 12090466 TI - Nutrition as a mediator in the relation between oral and systemic disease: associations between specific measures of adult oral health and nutrition outcomes. AB - Recent associations between oral health and systemic disease have led to renewed interest in the mouth and its contribution to health outcomes. Many pathways for this relationship have been postulated, among them the potential mediating role of nutrition. The link between various nutrients and systemic disease has been established, but relatively little work has been done in relating oral conditions with nutrition. We searched MEDLINE, from 1966 to July, 2001, to identify articles relating specific oral measures to nutrition outcomes. We included original articles written in English with a sample size greater than 30 that used objective oral health measures. We reviewed a total of 56 articles. Only a small proportion of these studies were methodologically sound. Although many studies were small and cross-sectional, the literature suggests that tooth loss affects dietary quality and nutrient intake in a manner that may increase the risk for several systemic diseases. The impact of tooth loss on diet may be only partially compensated for by prostheses. To date, there is little information relating periodontal disease and oral pain and nutrition. A few studies suggest poorer nutrition among individuals with xerostomia and altered taste. Further, impaired dentition may contribute to weight change, depending on age and other population characteristics. There is a paucity of well-designed studies addressing oral health and nutrition. Before we can acquire a better understanding of how nutrition and oral health interrelate, however, more studies will be required to confirm these associations-preferably longitudinal studies with larger sample sizes and better control of important confounders. PMID- 12090465 TI - Phospholipids in amelogenesis and dentinogenesis. AB - Phospholipids have been identified in enamel and dentin. Before demineralization, a group of phospholipids extracted by lipid solvents was associated with cell membranes and is therefore closely related to cell growth and intracellular regulations. After demineralization, a second group of phospholipids, associated with the extracellular matrix, was extracted; this group is probably linked to the mineralized phase. Using imidazole-osmium tetroxide fixation of rat incisors, we stained cellular unsaturated fatty acids, so that we could visualize the membrane domains, coated pits, and endocytic inclusions. Filipin, a probe for cholesterol, varied in density along the plasma membrane of secretory ameloblasts, and allowed us to visualize membrane remnants inside the forming enamel. With respect to phospholipids located in the extracellular matrix, the malachite-green-glutaraldehyde (MGA) method or iodoplatinate (IP) reaction retains and visualizes enamel and dentin phospholipids. In predentin, aggregates appearing as granules and filaments, or liposome-like structures, were located in the spaces between collagen fibrils. In dentin, organic envelopes coating the crystals, also named "crystal-ghost" structures, outlined groups of collagen fibrils. Histochemical data provided evidence that phospholipids are co distributed or interact with proteoglycans. Radioautography after IP reaction established that [3H] choline was detected in dentin as early as 30 min after the intravenous injection of the labeled precursor, before any labeling was seen in odontoblasts and predentin. This suggests that blood-serum-labeled phospholipids pass between odontoblasts, cross the distal permeable junctional complex, and diffuse in dentin prior to any cellular uptake and phospholipid synthesis. Pharmacologically and genetically induced pathology also supports the suggestion that phospholipids play an important role in the formation and mineralization of dental tissues. PMID- 12090467 TI - The role of matrix metalloproteinases in squamous cell carcinomas of the head and neck. AB - The ability of tumors to infiltrate the surrounding tissue is one of the major characteristics of a malignancy. This process is based on the tumors ability to destroy the extracellular matrix (ECM) including the basement membrane (BM). Several previous studies identified matrix metalloproteinases (MMPs), a family of zinc-dependent endopeptidases to be key players in this process. Since then multiple investigations focused on the expression and activation levels of their extracellular regulators and to a lesser extent of their transcriptional modulators. However, the exact diagnostic and prognostic values of these regulators still remain unclear. Squamous cell carcinomas of the head and neck (HNSCC) are known for their infiltrative growth and there is strong evidence that at least some members of the MMP-family play a crucial role in this process. It turned out that MMP-2, -9, -13 and to a lesser extent MMP-7 are related to the metastatic potential of HNSCC but further studies will be required to establish the exact role of MMPs in HNSCC. This Review will discuss the current literature concerning the role of MMPs in HNSCC. PMID- 12090468 TI - Dissimilar invasive and metastatic behavior of vincristine and doxorubicin resistant cell lines derived from a murine T cell lymphoid leukemia. AB - Multidrug resistance (MDR) lines from a murine T-cell lymphoid leukemia were selected in increasing vincristine (VCR) or doxorubicin (DOX) concentrations. Surface markers were determined by flow cytometry in both resistant (LBR-V 160 and LBR-D 160) and sensitive (LBR-) cell lines. Results obtained revealed similar expression of CD25, CD24, CD8, CD4, C18 and CD44, while differences in binding to hyaluronic acid (HA) were found. LBR- and LBR-D 160 bound to HA only after phorbol ester (PMA) activation, while LBR-V160 failed to bind HA even after PMA treatment. Histopathological analysis disclosed that LBR-V160 was less invasive than LBR- and LBR-D160 cell lines. In vitro growth of cell lines analyzed by sulforhodamine-B uptake showed that doubling time for the three lines was 10.24 h (LBR-), 16.75 h (LBR-V160) and 16.29 h (LBR-D160). Mortality rate was determined after i.p. injection of 10(4) cells. Mice inoculated with LBR- died at 23 2.11) days, while those inoculated with LBR-V160 or LBR-D160 died at 41 (+/- 9.53) or 41 (+/- 4.96) days, respectively. Our results demonstrated that leukemic murine T cells cultured in the long-term presence of VCR or DOX not only presented changes in the resistance phenotype but also variations in their growth and metastatic pattern. PMID- 12090469 TI - GPI-specific phospholipase D mRNA expression in tumor cells of different malignancy. AB - Glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) is a highly specific enzyme whose only known substrate is the GPI anchor of cell surface proteins. GPI-PLD measurements, however, are technically difficult since the enzyme is expressed at low levels in cells and tissues, and serum contains large amounts of inactive, latent GPI-PLD interfering with protein-based assays. We have therefore developed a semi-quantitative RT-PCR method to measure mRNA expression of all known GPI-PLD isoforms in cells and tissues. In human ovarian cancer cell lines, GPI-PLD mRNA expression correlated with GPI-PLD enzyme activity and with the shedding of the GPI-anchored tumor and prognostic markers, urokinase receptor and CA125, from the cell surface. This supports a potential role for this enzyme in the generation of circulating prognostic markers in malignant tumors. Similarly, in human epithelial cells of the skin, GPI-PLD mRNA expression increased with tumor progression. Whereas normal keratinocytes did not express significant amounts of GPI-PLD mRNA, expression was dramatically induced by serum in immortalized HaCaT keratinocytes and constitutively high and independent of serum in tumorigenic A431 epidermoid carcinoma cells. In addition, GPI-PLD expression was significantly increased in highly malignant. H-ras transfected murine bladder carcinoma cells as compared to the low malignant, non transfected parental cells. The competitive RT-PCR described here represents the first quantitative assay specific for cellular GPI-PLD isoforms, and our in vitro analyses suggest that GPI-PLD expression might be associated with tumor malignancy. PMID- 12090471 TI - Quantitative determination of tumor cell intravasation in a real-time polymerase chain reaction-based assay. AB - Tumor cells acquire the ability to enter blood vessels surrounding the primary tumor, endowing them with the capacity to disseminate and become established in distant sites, originating a metastasis. Determination of the intravasation ability of tumor cells is thus important, as it can be correlated with their potential malignancy. To analyze the intravasation phenotype of human tumor cells in vivo, we performed chick embryo chorioallantoic membrane (CAM) assays. Cells were inoculated on the CAM of 9-day-old chick embryos and the membrane at the opposite side of the egg was recovered after 48 h incubation. To measure intravasation ability, we calculated the amount of human DNA in each CAM sample by real-time PCR of Alu sequences and SYBR Green 1 fluorescence detection. This analysis showed a detection limit of 1 human cell per 10(5) total cells, and we were able to distinguish between tumor cells of distinct invasive capacity. This assay has several advantages over current methods to measure intravasation ability, including the elimination of post-PCR analysis, sensitivity and easy scale-up of sample numbers. PMID- 12090470 TI - Mitogen activated protein kinase pathway is involved in RhoC GTPase induced motility, invasion and angiogenesis in inflammatory breast cancer. AB - Inflammatory breast cancer (IBC) is the most lethal form of locally advanced breast cancer known. IBC carries a guarded prognosis primarily due to rapid onset of disease, typically within six months, and the propensity of tumor emboli to invade the dermal lymphatics and spread systemically. Although the clinical manifestations of IBC have been well documented, until recently little was known about the genetic mechanisms underlying the disease. In a comprehensive study aimed at identifying the molecular mechanisms responsible for the unique IBC phenotype, our laboratory identified overexpression of RhoC GTPase in over 90% of IBC tumors in contrast to 36% of stage-matched non-IBC tumors. We also demonstrated that overexpression of RhoC GTPase in human mammary epithelial (HME) cells nearly recapitulated the IBC phenotype with regards to invasion, motility and angiogenesis. In the current study we sought to delineate which signaling pathways were responsible for each aspect of the IBC phenotype. Using well established inhibitors to the mitogen activated protein kinase (MAPK) and phosphatidylinositol-3 kinase (PI3K) pathways. We found that activation of the MAPK pathway was responsible for motility, invasion and production of angiogenic factors. In contrast, growth under anchorage independent conditions was dependent on the PI3K pathway. PMID- 12090472 TI - Identification and validation of metastasis-associated proteins in head and neck cancer cell lines by two-dimensional electrophoresis and mass spectrometry. AB - Despite improvements in treatment of patients with head and neck squamous cell carcinoma (HNSCC) over the last two decades, the survival rate of these patients has not increased significantly. One of the major factors in the poor outcome of the disease is regional metastasis. To better understand the mechanisms of this process at the protein level, we performed two-dimensional electrophoresis (2-DE) and mass spectrometry using SELDI ProteinChip technology to identify proteins differentially expressed in two HNSCC cell lines, UMSCC10A and UMSCC10B, from the same patient. UMSCC10A was derived from the primary tumor and UMSCC10B from a metastatic lymph node. The differentially expressed proteins were excised from the gels. Following in-gel digestion by trypsin, mass profiles of the peptides were generated. Proteins were identified by submitting the peptide mass profiles to a public available NCBInr databases (www.proteometrics.com). Two membrane associated proteins, annexin I and annexin II and glycolytic protein enolase alpha were found to be upregulated, and calumenin precursor down-regulated, in metastatic cell line UMSCC10B. The identity of these proteins was confirmed by analyzing additional peptide mass fingerprints obtained by endoproteinase lysine C digestion. The results were also validated by Western blotting analysis. Our results showed that enolase-alpha, annexin-I and annexin-II might be important molecules in head and neck cancer invasion and metastasis. The results also suggest an important complementary role for proteomics in identification of molecular abnormalities important in cancer development and progression. PMID- 12090473 TI - TNFalpha and IL-8 regulate the expression and function of CD44 variant proteins in human colon carcinoma cells. AB - The mechanisms underlying the inflammatory and metastatic processes share a number of similar pathways, such as those involving adhesion, migration and extravasation. In this article, the effects of pro-inflammatory cytokines on metastatic-related activities of colon cancer cells were tested. The expression and biological activity of the proteoglycan CD44 in low (LS174T) and high metastatic (HM7) cell lines following exposure to TNFalpha and IL-8 were assessed. Treated cells expressed more CD44 splice variants (CD44v), while CD44 standard protein (CD44s) expression remained unchanged. Treatment with TNFalpha induced IL-8 secretion and IL-8 gene transcription in a time-dependent manner. Both cytokines enhanced the ability of the cells to adhere to the CD44-specific ligand hyaluronic acid, an effect that was specifically blocked by an anti-IL-8 antibody. These results suggest that the effect of TNFalpha on IL-8 is responsible for the regulation of the expression of CD44 isoforms. Additional experiments showed that neither of the cytokines tested regulate the expression of CD44 gene regulation via activation of a well-characterized specific 22-bp epidermal growth factor regulatory element present in the CD44 promoter sequence, suggesting that this is not the mechanism of activation. We conclude that immuno modulatory mediators can modify the expression of cell-to-cell or cell-to-matrix adhesion proteins, implicated in the determination of phenotypes associated with aggressiveness and metastasis of colon cancer cells. PMID- 12090475 TI - Optically imageable metastatic model of human breast cancer. AB - We report an optically imageable orthotopic metastatic nude mouse model of the human breast cancer MDA-MB-435 expressing green fluorescent protein (GFP). We demonstrate fluorescent imaging of primary and metastatic growth in live tissue and in intact animals. Fragments of tumor tissue expressing GFP were sutured into the pocket in the right second mammary gland. Tumor tissue was strongly fluorescent, enabling whole-body imaging of tumor growth by week 5. Neovascularization of the primary tumor was also visualized by whole-body imaging by contrast of the vessels to the fluorescent tumor. At autopsy, the MDA-MB-435 GFP was found to have metastasized to various organs, including the lung in 55% of the animals, the lymph nodes in 15% of the animals including axillary nodes, and the liver in 10% of the animals. These metastases could be visualized in fresh tissue by fluorescent imaging. Detailed fluorescence analysis visualized extensive metastasis in the thoracic cavity and the lymphatic system. Large metastatic nodules in the lung involved most of the pulmonary parenchyma in all lobes. Lymph node metastasis was found mainly in the axillary area. In the liver, fluorescent macroscopic metastatic nodules were found under the capsule. The metastatic pattern in the model thus reflected clinical metastatic breast cancer and provides a powerful model for drug discovery for this disease. PMID- 12090474 TI - Non-glycosylated tandem repeats of MUC1 facilitate attachment of breast tumor cells to normal human lung tissue and immobilized extracellular matrix proteins (ECM) in vitro: potential role in metastasis. AB - MUC1 is a transmembrane glycoprotein abundantly expressed on the apical surface of human ductal epithelial cells and over entire cell surface of tumors originating from those cells. It is 300 to 500 nm long and has a rigid, rod-like structure protruding from the cell surface. MUC1 expressed by normal cells has heavily O-glycosylated tandem repeat domain while MUC1 on malignant cells is aberrantly O-glycosylated. Substantially reduced (aberrant) glycosylation of the tandem repeat region of tumor MUC1 results in uncovering of the polypeptide core. This new structural feature may play an important role in the attachment of metastasizing tumor cells to tissues at distant sites. We show that MDA-MB-231 cells attaching to the immobilized extracellular matrix proteins (ECM) are higher MUC1 expressers than those non-attaching and that the attachment is inhibited by the addition of non-glycosylated, MUC1 peptide. This 100 a.a. peptide composed of 5 tandem repeats from the tandem repeat domain mimics the forms of MUC1 found in ascites fluid of cancer patients. We also show that this synthetic form of MUC1 inhibited attachment of breast tumor cells to sections of normal human lung tissue and immobilized ECM. We did not find correlation between the expression of Tn (GalNAc-Ser/Thr) epitope and the ability of tumor cells to adhere to the immobilized ECM. These results indicate that the non-glycosylated form of MUC1 plays a role in the initial attachment of carcinoma cells to tissues at distant sites, which may facilitate establishment of metastatic foci. PMID- 12090476 TI - FHIT protein expression and its relation to apoptosis, tumor histologic grade and prognosis in colorectal adenocarcinoma: an immunohistochemical and image analysis study. AB - The FHIT gene, a member of the histidine triad family has been identified as a tumor suppressor gene. Molecular genetic approaches to determine alterations in the FHIT gene in colorectal cancers have produced varying results with reported abnormalities of the FHIT gene transcripts in 13% to 50% of cases studied. FHIT has been reported to be involved in the regulation of apoptosis and cell cycle in cell culture systems. Immunohistochemical (IHC) studies of FHIT expression in human colon cancer and its correlation to apoptosis and clinical prognosis have been sparse. We studied 100 human colorectal cancers by IHC and a computerized image analysis (CIA) method to evaluate FHIT expression and the rate of apoptosis in tumors and corresponding mucosae. Follow-up data for at least five years was available for all patients. We correlated the results with tumor grade, stage and clinical prognosis. We used commercially available polyclonal anti FHIT antibody and Apoptaq kit on paraffin-embedded tumors and their corresponding mucosae and the SAMBA 4000 CIA system to evaluate the labeling index (LI), the mean optical density (MOD) of the stain and calculate a quick score (QS). The LI is the ratio of positively stained areas to the total area of the tissues examined, the MOD represents the concentration of the positive stain as measured per positive pixels and the QS a numeric product of the LI and MOD for each microscopic area examined. Image analysis of IHC staining of the tumor sections defined three main groups based on the reactivity of the anti FHIT polyclonal antibody. Group I included 23 cases where the LI was less than 55% with a mean of 36%. Eight cases in this group showed complete loss of FHIT expression. Group II included 41 cases where the LI was between 55% and 65% with a mean of 60%. Group III was composed of 36 cases where the LI was more than 65% with a mean of 69%. Our data showed that the absence or reduction of FHIT protein in the tumors, relative to morphologically normal mucosa, plays a role in the development of a few colorectal cancers (23%). Poorly differentiated carcinomas showed absent or decreased FHIT. A reduction of FHIT was positively correlated with the rate of distant metastases and worse prognosis. Over-expression of FHIT is directly proportional to the apoptotic rate in the tumors examined. CIA provides an objective and accurate assessment of the staining patterns and generates numerical data evaluating intensity better than depending on subjective light microscopy alone. PMID- 12090477 TI - Establishment of fluorescent lung carcinoma metastasis model and its real-time microscopic detection in SCID mice. AB - Lung cancer is the most prevalent malignant tumor in the world. Metastasis of the disease causes death in lung cancer patients. Recent study has shown that multiple cascades of gene defects occur in lung cancer. In this report, we established a novel H1299/EGFP tumor model to determine whether H1299 transfected with the enhanced green fluorescent protein (EGFP) gene in vitro and xenotransplanted into SCID mouse lung would permit the detection of lung cancer micrometastasis in vivo. We demonstrated that EGFP-transduced H1299 cells maintained stable high-level EGFP expressions during their growth in vivo. EGFP fluorescence clearly demarcated the primary seeding place and readily allowed for the visualization of distant micrometastasis and local invasion at the single cell level. Small metastatic and locally invasive foci, including those immediately adjacent to the tumor's leading invasive edge, were almost undetectable by routine hematoxylin and eosin staining and immunohistochemistry. The GFP tagged lung cancer model is superior for the detection and study of physiologically relevant patterns of lung cancer invasion and metastasis in vivo. PMID- 12090478 TI - Stability of attachment representations: the transition to marriage. AB - This study examined the stability of adult attachment representations across the transition to marriage. One hundred fifty-seven couples were assessed using the Adult Attachment Interview (AAI; C. George, N. Kaplan, & M. Main, 1985), the Current Relationship Interview (J. A. Crowell & G. Owens, 1996), and measures describing relationship functioning and life events 3 months prior to their weddings and 18 months into their marriages. The authors tested the hypotheses that attachment classifications are stable and that change is related to experiences in the relationship and/or life events; 78% of the sample received the same primary AAI classification (secure, preoccupied, and dismissing) at both times. Change was toward increased security and was associated with feelings and cognitions about the relationship. Only 46% of participants initially classified as unresolved retained the classification. Stability of the unresolved classification was associated with stressful life events and relationship aggression. PMID- 12090479 TI - A longitudinal analysis of friendships and substance use: bidirectional influence from adolescence to adulthood. AB - The reciprocal relation between deviant friendships and substance use was examined from early adolescence (age 13-14) to young adulthood (age 22-23). Deviance within friendships was studied using direct observations of videotaped friendship interaction and global reports of deviant interactions with friends as well as time spent with friends. Substance use was assessed through youth self report at all time points. Multivariate modeling revealed that substance use in young adulthood is a joint outcome of friendship influence and selection processes. In addition, substance use appears to influence the selection of friends in late adolescence. Findings suggest that effective preventions should target peer ecologies conducive to substance use and that treatment should address both the interpersonal underpinnings and addiction processes intrinsic to chronic use, dependence, and abuse. PMID- 12090480 TI - Contextual determinants of mothers' interventions in young children's peer interactions. AB - In this study, the role of context in mothers' interventions in their preschool children's peer relationship problems was investigated. Event theme (aggression, peer rebuff, or initiating play), the child's role in the event (actor or target), the child's age and sex, and the mother's emotional reaction were examined as predictors of the extent to which mothers (N = 71) said they would discuss peer relationships, encourage peer interaction, and use power assertion in response to a series of videotaped vignettes depicting common peer relationship problems. Mothers suggested using more discussion in aggressive situations and more encouragement in initiating play situations. Mothers said they would use more power assertion when the child was the actor (i.e., provocateur), rather than the target, in an aggressive situation. Findings are discussed in terms of (a) the importance of considering context in understanding how mothers intervene in their children's peer relationships and (b) the need to examine moderators of cross-contextual consistency in mothers' interventions. PMID- 12090481 TI - Matching and naming objects by shape or function: age and context effects in preschool children. AB - Three experiments tested preschoolers' use of abstract principles to classify and label objects by shape or function. Three- and 4-year-olds were instructed to match objects by shape or function. Four-year-olds readily adopted either rule, but 3-year-olds followed only the shape rule. Without a rule, 4-year-olds tended to match by shape unless object function was shown during matching (Experiment 2). Three-year-olds' ability to use a function rule was tested in several conditions (re-presenting functions; reminders to "use the rule"; repeating rule on every trial). None induced consistent function matching (Experiment 3). Supplemental memory and verbal tasks showed that 3-year-olds have trouble using function as an abstract basis of comparison. Naming data, however, show that preschoolers are learning that object labels are based on function. The results show preschoolers' growing flexibility in adopting abstract generalization rules and growing knowledge of conventions for extending words. PMID- 12090482 TI - Children's competence and value beliefs from childhood through adolescence: growth trajectories in two male-sex-typed domains. AB - The purpose of this study was to document gender differences in children's competence and value beliefs (N =514) from the 1st through 12th grades and to investigate the relation of these trends to initial differences in parents' perceptions of children's ability. Six separate growth models were tested: math competence, math interest, math importance, sports competence, sports interest, and sports importance. Across all 6 models, children's self-perceptions declined from 1st grade to 12th grade. Gender differences in competence and value beliefs were found. The gap between boys' and girls' competence beliefs decreased over time. In addition, parents' initial ratings of children's ability helped to explain mean level differences and variations in the rate of change in children's beliefs over time, with the effect being strongest in the sports models. PMID- 12090483 TI - Mothers' insightfulness regarding their infants' internal experience: relations with maternal sensitivity and infant attachment. AB - This study examined the associations among mothers' insightfulness into their infants' internal experience, mothers' sensitivity to their infants' signals, and infants' security of attachment to their mothers. The insightfulness of 129 mothers of 12-month-old infants was assessed by showing mothers 3 videotaped segments of observations of their infants and themselves and interviewing them regarding their infants' and their own thoughts and feelings. Interviews were classified into 1 insightful and 3 noninsightful categories. Mothers' sensitivity was assessed during play sessions at home and at the laboratory, and infant mother attachment was assessed with the Strange Situation. Mothers classified as positively insightful were rated as more sensitive and were more likely to have securely attached children than were mothers not classified as positively insightful. Insightfulness also accounted for variance in attachment beyond the variance explained by maternal sensitivity. These findings add an important dimension to research on caregiving, suggesting that mothers' seeking of explanations for the motives underlying their infants' behavior is related to both maternal sensitivity and infant attachment. PMID- 12090484 TI - The cognitive profile and multiple-deficit hypothesis in Chinese developmental dyslexia. AB - The present study was conducted to examine the cognitive profile and multiple deficit hypothesis in Chinese developmental dyslexia. Thirty Chinese dyslexic children in Hong Kong were compared with 30 average readers of the same chronological age (CA controls) and 30 average readers of the same reading level (RL controls) in a number of rapid naming, visual, phonological, and orthographic tasks. Chinese dyslexic children performed significantly worse than the CA controls but similarly to the RL controls on most of the cognitive tasks. The rapid naming deficit was found to be the most dominant type of cognitive deficit in Chinese dyslexic children. Over half of the dyslexic children exhibited deficits in 3 or more cognitive areas, and there was a significant association between the number of cognitive deficits and the degree of reading and spelling impairment. The present findings support the multiple-deficit hypothesis in Chinese developmental dyslexia. PMID- 12090485 TI - Perception of the symmetrical patterning of human gait by infants. AB - Two experiments tested 3- and 5-month-old infants' sensitivity to properties of point-light displays of human gait. In Experiment 1, infants were tested for discrimination of point-light displays of a walker and a runner, which, although they differed in many ways, were equivalent with regard to the phasing of limb movements. Results revealed that 3-month-old, but not 5-month-old, infants discriminated these displays. In Experiment 2, the symmetrical phase-patterning of the runner display was perturbed by advancing two of its limbs by 25% of the gait cycle. Both 3- and 5-month-old infants discriminated the walker display from this new phase-shifted runner display. These findings suggest that 3-month-old infants respond to the absolute and relative motions within a single limb, whereas 5-month-old infants respond primarily to the relations between limbs and, in particular, to the bilateral symmetry between the limbs. PMID- 12090486 TI - Differences in early parent-child conversations about negative versus positive emotions: implications for the development of psychological understanding. AB - The authors examined whether the quality and content of everyday parent-child conversations about negative emotions are the same or different from everyday talk about positive emotions. Extensive longitudinal speech samples of 6 children and their parents were analyzed for several critical features when the children were between 2 and 5 years of age. Results showed that children and parents talked about past emotions, the causes of emotions, and connections between emotions and other mental states at higher rates during conversations about negative emotions than during conversations about positive emotions. Discourse about negative emotions also included a larger emotion vocabulary, more open ended questions, and more talk about other people. These differences appeared before the children's 3rd birthdays and remained consistent through the preschool years. The findings strengthen and clarify current understanding of young children's articulation and knowledge about people's minds, lives, and emotions. PMID- 12090487 TI - Two-year-olds' search strategies and visual tracking in a hidden displacement task. AB - Children younger than 3 years have difficulty with search tasks that involve hidden displacement. Partial visual information was provided about a ball's path as it moved toward a hiding place. Children (2.0 and 2.5 years old) saw a ball rolling down a ramp placed behind a transparent screen with 4 opaque doors. A wall, placed on the ramp and directly behind 1 of the doors, protruded above the screen and stopped the ball. Children were asked to find the ball. The transparency of the screen permitted visual tracking of the ball between the doors, but its final resting place was obscured. Both age groups were equally proficient at tracking the ball as it rolled behind the screen, but the 2.5-year olds were more likely to reach to the correct door. Looking behavior was related to errors in the younger group in that tracking that stopped short or continued past the correct door was associated with incorrect choices. PMID- 12090488 TI - The antecedents and correlates of agreeableness in adulthood. AB - Data from a 25-year prospective study of 194 individuals indicated that teacher and peer reports of aggression, compliance, and self-control at age 8 distinguished high-agreeable from low-agreeable adults at age 33. Profile analyses revealed two behavioral types in childhood and two personality types in adulthood, with considerable continuity in the composition of these high- and low agreeable types over time. High-agreeable childhood types had fewer disobedience and concentration problems than low-agreeable childhood types, and among boys, high-agreeable childhood types had better school grades and fewer behavior problems than their low-agreeable counterparts. High-agreeable adulthood types reported less alcoholism and depression, fewer arrests, and more career stability than did low-agreeable adulthood types. PMID- 12090489 TI - Young children can extend motion verbs to point-light displays. AB - In the first study using point-light displays (lights corresponding to the joints of the human body) to examine children's understanding of verbs, 3-year-olds were tested to see if they could perceive familiar actions that corresponded to motion verbs (e.g., walking). Experiment 1 showed that children could extend familiar motion verbs (e.g., walking and dancing) to videotaped point-light actions shown in the intermodal preferential looking paradigm. Children watched the action that matched the requested verb significantly more than they watched the action that did not match the verb. In Experiment 2, the findings of Experiment 1 were validated by having children spontaneously produce verbs for these actions. The use of point-light displays may illuminate the factors that contribute to verb learning. PMID- 12090490 TI - Are parents' gender schemas related to their children's gender-related cognitions? A meta-analysis. AB - Meta-analyses were conducted of 43 articles (with 48 different samples) investigating the relationship between parents' gender schemas and their offspring's gender-related cognitions. The parents' offspring ranged in age from infancy to early adulthood. Offspring measures included gender self-concept, gender attitudes toward others, gender-related interests, and occupational attitudes. Overall, a small but meaningful effect size (r = .16) indicated a significant and positive correlation between parent gender schemas and offspring measures. Specifically, parents with more traditional gender schemas were more likely than parents with more nontraditional schemas to have offspring with gender-typed cognitions about themselves or others. In addition, the magnitudes of observed effect sizes were influenced by particular moderator variables, including type of parent gender schema (gender self-concept vs. gender attitudes toward others), type of offspring gender-related cognitions, parent gender, offspring gender, offspring age, and publication characteristics. The results are cautiously interpreted as suggesting a possible influence of parents on the development of their children's gender-related thinking. PMID- 12090491 TI - Manic-depressiveness, obsessive-compulsive tendencies, and suicidality in Kuwaiti college students. AB - In a sample of 87 Kuwaiti undergraduates (20 men, 67 women), obsessive-compulsive scores were associated with scores on measures of mania, depression, and suicidal thoughts. Multiple stepwise regression indicated that (a) obsession, suicidal attempts, and suicidal threats were predictors of depression, (b) depression, suicidal attempts, and obsession predicted suicidal thoughts, (c) age, doubting, and sex were predictors of suicidal threats, and (d) suicidal thoughts, depression, and obsession were predictors of suicidal attempts. PMID- 12090493 TI - Parental psychological control and attachment in late adolescents and young adults. AB - The aim of this study was to examine the association between parental psychological control and parental attachment. Another goal was to investigate the differential and combined effects of these two constructs on indices of psychological well-being, namely, self-esteem, positive or negative affect, and personal and interpersonal efficacy. Subjects were 319 males (n = 142) and females (n = 177) with an age range of 15 yr., 9 mo. to 22 yr., 5 mo. (M = 20.0, SD = 1.6). Analysis showed that psychological control was negatively related to attachment. Both psychological control and attachment predicted self-esteem, and there was some indication that psychological control exercised by mothers had a stronger influence on self-esteem. Attachment was also a significant predictor of positive and negative affect and perceived personal efficacy. No sex or age differences were found. PMID- 12090492 TI - Cross-sectional analysis of intimacy, passion, and commitment: testing the assumptions of the triangular theory of love. AB - Using data from two previous studies, the current study tested the assumptions in the Triangular Theory of Love about changes in intimacy, passion, and commitment over time. Using a cross-sectional design, the study examined differences in the three components among 446 romantically involved individuals who were either casually dating, exclusively dating, engaged, or married. In support for the assumptions in the Triangular Theory specifically, analyses indicated significant negative partial correlations between intimacy and relationship length as well as between passion and relationship length. The correlation between commitment and relationship length was significant and positive. One-way analysis of variance of relational stage gave similar results. Reported intimacy and passion scores were lowest for participants who were casually dating, higher for participants who were engaged, and lower for married participants. Reported commitment scores increased from casually dating participants to the married participants. PMID- 12090494 TI - Age and sex differences for anxiety in relation to family size, birth order, and religiosity among Kuwaiti adolescents. AB - Differences in rated anxiety among 2,453 boys (n= 1,229) and girls (n = 1,224), Kuwaiti secondary school students, were reported for five age groups from 14 to 18 years. For girls at all ages but 14 years, mean rated anxiety was significantly higher than the means for the boys. Mean anxiety scores increased across age groups from 14 to 18 years. Not all comparisons between age groups with the same sex, however, were significant. Analysis showed nonsignificant correlations for anxiety with both family size and number of siblings, but significant and positive correlations for anxiety with birth order were found for boys (r=.10, p<.01) and girls (r=.06, p<.05). The predictive and practical values of these very small correlations are negligible, being significant merely because N is so large. Pearson correlations between anxiety and self-rating of religiosity were -.22 and -.22 (p<.01) for boys and girls, respectively. This result was interpreted in the light of high intrinsic religious orientation among Kuwaitis. In the Islam proper, multiple practices are said to relieve anxiety. PMID- 12090495 TI - Exploratory studies of classical conditioning of the preoral cavity in harnessed carpenter ants (Camponotus pennsylvanicus). AB - An attempt was made to classically condition the mouthparts of harnessed worker ants (Camponotus pennsylvanicus) in anticipation of feeding. Experiments were designed to investigate classical conditioning with one CS, discrimination between two CSs, and pseudoconditioning. Analysis indicated a small acquisition effect that could be accounted for by pseudoconditioning. The preparation can be used to study nonassociative learning and some instrumental conditioning situations. PMID- 12090496 TI - Scholarly productivity of editors of social work and psychology journals. AB - This study explored the scholarly productivity of editors of selected social work and psychology journals. Analysis indicated that editors of psychology journals had statistically significantly greater scholarly achievement than editors of social work journals. These findings suggest that scholarly achievement as measured appears to be of less importance in appointing editors to the five selected social work journals than in appointing editors to the five selected psychology journals. PMID- 12090497 TI - An English language version of the social support list: preliminary reliability. AB - The 34-item Social Support List developed in The Netherlands combines measures of support satisfaction and support interactions in six subscales plus a total score. The present study was designed to assess the applicability of the list in a different cultural context. Data from 421 American undergraduates were consistent with Dutch findings and support the efficacy of the English language version. PMID- 12090499 TI - The 'troubles' in Northern Ireland and suicide. AB - In Northern Ireland from 1965 to 1997, suicide rates and deaths from the civil unrest were negatively associated. PMID- 12090498 TI - Pyridostigmine bromide and the long-term subjective health status of a sample of over 700 male Reserve Component Gulf War era veterans. AB - Data from a 1996-1997 survey of approximately 700 Reserve Component male veterans indicate that the consumption of pyridostigmine bromide pills, used as a pretreatment for potential exposure to the nerve agent Soman, was a significant predictor of declines in reported subjective health status after the war, even after controlling for a number of other possible factors. Reported reactions to vaccines and other medications also predicted declines in subjective health. While higher military rank generally predicted better health during and after the war, educational attainment, minority status, number of days in theater, and age generally did not predict changes in subjective health. Although servicemembers were directed to take three pills a day, veterans reported a range of compliance- less than a fourth (24%) followed the medical instructions compared to 61% who took fewer than three pills daily and 6% who took six or more pills a day. Implications for use of pyridostigmine bromide are discussed. PMID- 12090500 TI - Reasoning about interpersonal responsibility when making judgments about scenarios depicting close personal relationships. AB - This study examined the moral reasoning of 36 young adults (M age = 20.8 yr., evenly divided by sex) about situations involving close interpersonal relationships: best friends, spouses, and parents or children. Participants were presented stories in which personal needs and desires were in conflict with interpersonal concerns. When asked how the conflicts should be resolved, the large majority of participants judged that personal desires should be subordinated to interpersonal concerns, justifying their judgments with reference to concern for others, relationship maintenance, or relationship responsibilities. No sex differences in judgments or justifications were found. In addition, most judged that the act of meeting another's needs in a close relationship was an obligatory responsibility and that such responsibilities generalized to people living in other societies. Thus, participants considered interpersonal responsibilities to be moral obligations that apply across cultural boundaries and did not consider them to be matters of personal choice. PMID- 12090501 TI - Tomboys, masculine characteristics, and self-ratings of confidence in career success. AB - Previous research focusing on gender-roles and self-concept suggests a positive relationship between masculinity and self-concept. The present study explores self-perceptions of being a "tomboy," gender-role and ratings of self-confidence in later success. Higher self-ratings on the tomboy scale correlated with confidence in career success but not with confidence in passing a college course or jogging for 20 min. There was a positive correlation between masculinity scores but not femininity scores with tomboy ratings. Masculine characteristics such as independence, aggression competitiveness, leadership ability, ability to defend, stand by beliefs, mediated by compassion appear to be positively correlated with perceptions of success. PMID- 12090502 TI - Posttraumatic stress and depressive symptomatology in a sample of battered women from South Africa. AB - This study investigated the incidence of depression, posttraumatic stress symptomatology, anger, and guilt in a sample of 40 battered women in a shelter. Analysis indicated that 63% of the 40 scored moderate to severe on depression, 59% manifested high posttraumatic stress symptoms, while 38.4% and 54.5% reported problems with anger and guilt, respectively. Frequency and severity of physical abuse were not correlated with depressive or posttraumatic stress symptomatology. PMID- 12090503 TI - Shortening a quality of life questionnaire for adolescents with chronic pain and its psychometric qualities. AB - Chronic pain is a common experience in adolescence. To measure its influence on quality of life in adolescents with chronic headache, Langeveld developed the 71 item scale, Quality of Life Headache-Youth. On the basis of this questionnaire we develop a shortened list, also suitable for other pain locations to enhance compliance. For this, we tested a sample of 98 adolescents from an open population with chronic benign pain. This article presents the psychometric qualities of the shortened version, named the Quality of Life Questionnaire for Adolescents with Chronic Pain. The original version could be reduced to 44 items which showed suitable internal consistency and construct validity against COOP/WONCA charts. PMID- 12090504 TI - Relationships of personality disorders with MMPI-2 malingering, defensiveness, and inconsistent response scales among forensic examinees. AB - MMPI-2 validity scales were correlated with MCMI-II personality disorder scales to examine relationships between response styles and personality disorders in a sample of 84 criminal defendants. 14 MMPI-2 validity scales were significantly correlated with 13 personality disorders. All of the personality disorder scales were significantly correlated with at least one validity measure and 11 of 13 personality disorder scales were significantly correlated with two or more MMPI-2 validity scales. While a personality disorder diagnosis may have a general effect on validity scales, relationships theoretically consistent with a given personality disorder were also found. This means that response set appears to be a manifestation of personality, and as such, examiners should expect symptom amplification or minimization or inconsistent responses, based on an individual's personality. Subsequently, forensic examiners are encouraged to evaluate the relationships between MCMI-II personality disorders and MMPI-2 validity scales to avoid misjudging MMPI-2 profiles as invalid when they accurately reflect manifestations of personality. PMID- 12090505 TI - Profiles of learning disability subtypes in a retrospective sampling of adolescents' scores on the Woodcock-Johnson Psycho-Educational Battery-Revised. AB - This study was an effort to subtype adolescents' learning disabilities using scores on the Woodcock-Johnson Psycho-Educational Battery-Revised. A cluster analysis of 88 cases classified as learning disabled and 75 as nonlearning disabled was conducted to characterize subtypes. Two distinct subtypes emerged for the group while one for the nonlearning disabled group was identified. A multivariate analysis of variance to test for differences in scores among the identified subtypes yielded significant differences, suggesting that these students had two specific types of learning disorders. Cross-validation is essential. PMID- 12090506 TI - Commitment and exploration as mechanisms of identity formation. AB - We report a two-wave longitudinal study of 1,571 Dutch adolescents concerning the role of commitment and exploration in identity development. We used the Utrecht Groningen Identity Development Scale to measure commitment and exploration in the domains of relational and societal identity. Our results can be summarized in three points. (1) Commitment and exploration are related processes in the development of identity. Adolescents with strong commitments also frequently explore them, and adolescents with low exploration in general have weak commitments. (2) The longitudinal stability of commitment and exploration has a medium effect size. For relational identity the stability of commitment is greater than that of exploration, but this is not the case for societal identity. The explanation we give for the lack of this difference in stability between commitment and exploration in societal identity is that the formative period for societal identity comes primarily at the end of adolescence. In that connection, we conclude that for present-day Dutch adolescents the formation of relational identity probably precedes that of societal identity. (3) In neither identity domain is commitment predictive of exploration three years later, nor is the reverse the case. We conclude that no long-term developmental sequentiality of commitment and exploration was found, but the results do not rule out the possibility of short-term developmental sequentiality. PMID- 12090507 TI - Sex differences in the personal conceptions of intelligence: particularities of the Portuguese cultural context. AB - Several empirical studies have implicated helplessness as a problem in girls' lack of achievement, generally associated with static conceptions of intelligence. Yet, two studies aiming to explore sex differences in personal conceptions of intelligence, carried out with Portuguese samples of 222 and 2,106 subjects, respectively, using a 15-item rating scale to measure personal conceptions of intelligence adapted for Portuguese adolescents (11 to 17 years old), did not find sex differences, suggesting that certain specific characteristics of the Portuguese cultural context and its underlying socialization processes may have altered the expected kind of differentiation. Researchers should investigate whether success is not stereotypically attributed to male role norms and the importance of arousal of standards of achievement for women in the Portuguese culture. PMID- 12090508 TI - Development of a law enforcement stress and coping questionnaire. AB - This paper describes the construction of a self-report checklist designed to examine experiences of stressful routine occupational events and life events, coping responses and psychological distress among law enforcement officers. The checklist items were derived from existing scales and are designed to examine a cognitive phenomenological model of stress and coping and testing the goodness of fit hypothesis. PMID- 12090509 TI - Extended lowered body temperature increases the effective CS-US interval in conditioned taste aversion for adult rats. AB - Assuming body temperature correlates with metabolic activities, rate of body temperature recovery was manipulated to assess effects on long-trace conditioning in a conditioned taste-aversion paradigm. Following 10 min. access to a .1% saccharin solution and then 10 min. immersion in 0-0.5 degrees C water, two groups of 16 Wistar-derived, 81-113 day-old, male albino rats received either saline or lithium chloride injections 3 hr. later. These two groups were subdivided on basis of warming rate during the 3-hr. interval. Half of the rats recovered at room temperature (20 degrees to 21 degrees C), and half recovered in an incubator maintained at 30 degrees C. Maintaining a lowered body temperature between the conditioned stimulus and unconditioned stimulus allowed an association to be made at 3 hr., an interval that normally does not support conditioning. In contrast, lowering body temperature and then inducing a fast warming rate did not produce evidence of an aversion. It is suggested that maintaining a low body temperature over the interval between the presentation of the conditioned stimulus and unconditioned stimulus slows a metabolic clock that extends the measured interval at which associations can be made using conditioned taste-aversion procedures. PMID- 12090510 TI - Relation of career beliefs to career indecision among distance education adult college students in Taiwan. AB - This study constructed a predictive model of career indecision for adult college students participating in distance education by using a sample of 178 adult college students in Taiwan's Open University. The stepwise regression analysis of the career indecision scale, using sex, age, and 20 subscores of the Career Beliefs Checklist as predictor variables, yielded four variables (age and three major core career beliefs. Self-sufficiency, External Influences, and Self advocacy) as significantly contributing to the reported variance in the Career Indecision score of the Career Decision Scale. A Pearson product-moment correlation matrix described the relationships between the Career Indecision score and the 20 subscores of the Career Beliefs Checklist, age, and sex. Recommendations for incorporating career beliefs into career education courses are offered. PMID- 12090511 TI - Academic elite in accounting: linkages among top-ranked graduate programs. AB - The 10 top-ranked graduate programs in accounting, based on a national survey of deans and top administrators, were linked to one another by hiring in the programs one another's graduates. Almost one-half (45.9%) of the faculty members in these 10 programs (N = 172) had graduated from one of these 10 programs. It is suggested that this linkage helps these programs to maintain and enhance their prestige. PMID- 12090512 TI - Preliminary validity data on the Rasch Manic-Depressiveness Scale. AB - The 18-item Manic-Depressiveness Scale was revised via a Rasch top-down purification' procedure to provide a new 12-item scale with no significant age or sex bias. Application of this scale to previously collected data indicated that patients with manic-depression and schizophrenia receive comparable scores, which are significantly higher than those of a control group of students. Moreover, for a subgroup of patients with manic-depression, scores correlated .55 (p<.001) with number of manic-depression-relevant medications being taken. These findings lend support to the validity of the Rasch Manic-Depressiveness Scale as a general measure of psychoticism and psychiatric status. PMID- 12090513 TI - Various types of exercise and scores on the Beck Depression Inventory. AB - This study investigated scores on depression of students in sports. 311 (ns = 60, 100, 100, 40, 11) female athletic candidates were selected randomly and administered the Beck Depression Inventory. Initially, 60 persons who had gone through a body-building course for less than 1 month took the inventory and again after 4 wk. The mean score difference was -3.5. This decline in mean depression score after 4 wk. of exercise was significant. In the next step, 100 women new to body building and 100 women who had completed 2 mo. body building were compared. Their mean scores were 13.3 for newcomers and 10.2 for the 2-mo. group, a significant difference. Finally, 40 female athletes who had swum for less than 1 mo. were compared with professional swimmers from the Fars province. Their mean depression scores were 10.5 and 6.5, respectively. This difference was also significant. We concluded that exercising is associated with lower mean scores on the Beck Depression Inventory. PMID- 12090514 TI - Suggestions regarding cross-cultural environment as context for aging and human development in non-Western cultures. AB - An individual's aging and human development always occur within a articular cultural context, but given the dominance of Western culture, the environment of non-Western societies is cross-cultural--it includes not only a non-Western dimension, but also Global and Western dimensions. The Western dimension is made up of those characteristics associated with Western societies, which are now found in non-Western societies, for example, Western education, healthcare services, and indusrialization. The non-Western dimension, on the other hand, is made up of those attributes that are indigenous to the people, e.g., traditional healing. The Global dimension is characterized by standards that all societies should adhere to, e.g., human rights, equality for women, or by biological changes that are universally shared. Given this situation, six possible interaction patterns between the individual and his environment are mentioned as they have implications for different aging and human development pathways. These cross-cultural environmental dimensions also have implications for immigrants and refugees from non-Western societies in Western cultures and for ethnic minority group members in dominant cultures. This initial proposal may stimulate further detailed development. PMID- 12090515 TI - Measure of beliefs about improvements in mood associated with exercise. AB - Changes in mood following exercise are well documented; however, one particular limitation of research examining mood changes associated with exercise is that studies have not controlled for a possible expectancy effect. Given the difficulty associated with utilising a placebo control group in exercise settings and that no current questionnaires are available to measure beliefs concerning mood improvements during and after exercise, the present studies aimed to develop a suitable instrument for assessing these beliefs. In the first of these studies, 220 regular runners were respondents in developing the new scale. A logical approach to test construction, as proposed by Hase and Goldberg in 1967, produced two scales of acceptable reliability: a five-item subscale to measure beliefs concerning mood improvements during running (Cronbach alpha=.77) and a two-item subscale to measure beliefs about mood improvements following running (Cronbach alpha=.86). A second study, which involved the administration of these scales to 50 regular runners after a 60-min. treadmill run, indicated there were moderate associations between scores on these measures and mood changes during and after the run. PMID- 12090516 TI - Psychology of the scientist: LXXXIV: Raymond Bernard Cattell (1905-1998). AB - This obituary of R. B. Cattell (1905-1998) describes briefly some of the evidence underlying the judgment that Cattell was one of the half dozen greatest psychologists of the twentieth century and also portrays the gentle thoughtful man he was as a family member, friend, and colleague. PMID- 12090517 TI - Convergent validity of the Collett-Lester Fear of Death and Templer Death Anxiety Scales in Egyptian male college students. AB - The Collett-Lester Fear of Death Scale and the Templer Death Anxiety Scale were administered to 57 male Egyptian undergraduates. Pearson correlations between the total score on the Templer's scale and the four subscales of Collett-Lester's Fear of Self death, Self dying, Other's death, and Other's dying were .54, .55, .52, and .56, respectively, while the correlation between the total scores on the Collett-Lester and Templer scales was .73, denoting the convergent validity of the Collett-Lester scale against the Templer scale as criterion. PMID- 12090518 TI - A prospect theory explanation of the disposition to trade losing investments for less than market price. AB - Investors have a proven general reluctance to realize losses. The theory of "mental accounting" suggests that losses are easier to accept when mentally integrated with either preceding losses or with compensatory gains. Mental integration is made easier when a failed asset is exchanged against a new, apparently profitable, acquisition. The alternative is to sell the existing asset on the open market before re-investing the proceeds as desired. This is emotionally less appealing than "rolling over" a losing investment into a new venture by way of an asset trade. The psychological benefits of exchanging rather than selling a failed asset come at a cost. It is typical of trade-in arrangements, e.g., where one trades an old car against a new one, that the effective sale price of the existing asset is less than current market value. Acceptance of this low price adds to the investor's total monetary loss on the existing asset but is essential to an overall package deal apart from which that asset would often remain belatedly unsold. PMID- 12090519 TI - Sensitivity of the semantic fluency subtest of the repeatable battery for the assessment of neuropsychological status. AB - This study examined the concurrent validity of the Semantic Fluency subtest of the Repeatable Battery for the Assessment of Neuropsychological Status. Performance by cognitively impaired vs nonimpaired neurological patients was examined in relation to the performance of these groups on the Controlled Oral Word Association Test. Analyses indicated these tests are largely equivalent with respect to both sensitivity to neurocognitive dysfunction and discrimination of cognitively impaired from nonimpaired neurological patients. Findings support the psychometric validity of the Semantic Fluency subtest in the evaluation of brain dysfunction. PMID- 12090520 TI - Association of self-preoccupation and self-reported duration and severity of depressive episodes. AB - The present study investigated the relation of self-preoccupation, i.e., the tendency to focus more on the self than on external objects and to maintain this self-focused attention, with the duration and severity of depressive episodes. The hypotheses were that, compared with people low on self-preoccupation, people scoring higher on self-preoccupation may experience more long-lasting depressive episodes and during such episodes they may experience a greater number of depressive symptoms. The present study examined association of self-esteem with the severity or duration of depressive episodes. A total of 1,076 undergraduates (361 men, 700 women, 15 unknown) from nine universities in the Tokyo metropolitan area participated. Of the total, 246 undergraduates (55 men and 191 women) reported that they had experienced a depressive episode continuing four or more days; these data were analyzed and are reported here. Self-preoccupation, self esteem, and depressive episodes within 12 mo. were measured by a self-report questionnaire. The results tentatively supported the hypotheses. Specifically, rated self-esteem was significantly correlated with the number of depressive symptoms reported in a depressive episode but not with the duration of the episode. However, rated self-preoccupation was correlated with the severity and duration of the depressive episode, although the correlation of self preoccupation scale scores with the duration of the episode was quite weak and requires further research. PMID- 12090521 TI - Factorial validity of the Arabic Obsessive-Compulsive Scale in two cultures. AB - The Arabic Obsessive-Compulsive Scale and the Maudsley Obsessional Compulsive Inventory were administered to 87 Kuwaiti (Arabic version) and 73 American college students (English form). Pearson correlations for the total score on the Arabic scale and the four subscores of the Maudsley scale: Checking, Cleaning, Slowness, and Doubting were .72, .48, .50, and .61 for the Kuwaitis and .52, .40, .51, and .52 for the Americans. The loadings of the total score of the Arabic scale onto a factor of obsession and compulsion were .83 and .79 for Kuwaiti and American students, respectively, denoting a good factorial validity of the Arabic scale. PMID- 12090522 TI - Narcissism and anger: an exploration of underlying correlates. AB - Many have posited that narcissistic individuals are sensitive to insults and prone to react to such criticism with anger and hostility. Given that such anger is provoked by criticism from others, it was hypothesized that narcissistic traits which relate to perceived position of authority or superiority and an inflated sense of entitlement would make it easier for someone to perceive a remark as threatening and feel angry. It was hypothesized that these aspects of narcissism, as measured by the subscales of the Narcissistic Personality Inventory by Raskin and Hall would correlate differentially with anger, as measured by the Novaco Anger Scale in a sample of undergraduate men. Results partially confirmed the hypothesis and indicated that scores on Entitlement and Authority were positively associated with anger scores; however, superiority was not. The results imply that specific narcissistic traits such as a perceived position of authority and an inflated sense of entitlement, but not necessarily the global diagnosis of Narcissistic Personality Disorder, might be important to investigate as correlates of anger. PMID- 12090523 TI - Sleep, dreams, and job satisfaction. AB - Intercorrelations between the KJP Dream Inventory, Stanford Sleepiness Scale, and the Brayfield-Rothe Job Satisfaction Questionnaire from a database of 56 graduate students gave small significant correlations between job satisfaction and age (.32), repetitive traumatic dreaming, (-.30), being chased in one's dreams ( .35), and sleepiness (-.30). Associations of employment status with sleepiness, dream discontent, and dreams of being famous were also evident. Results were discussed in terms of the hypothesized relationship between contentedness or discontentedness and their association with sleep, dreaming, and job satisfaction. PMID- 12090524 TI - Reading comprehension among graduate students. AB - The present purpose was to examine graduate students' reading comprehension and reading vocabulary by comparing their scores on a standardized reading rest with scores obtained by a large normative sample of undergraduates. Participants were 59 graduate students from various disciplines, enrolled in three sections of an introductory educational research course at a southeastern university. These students were administered the Nelson-Denny Reading Test-Form G. Analysis showed these students had higher scores on the reading comprehension portion than did the normative sample of 5,000 undergraduate students from 38 institutions (Cohen d=.71). Also, the graduate students' scores on the reading vocabulary portion were higher (d=.45). PMID- 12090525 TI - Comparison of students' and faculty's perceptions of occurrence of dishonest academic behaviors. AB - Examination of estimates of prevalence of 28 dishonest academic behaviors provided by 217 students and 38 faculty members from the same Latin American institution shows faculty's perceptions differed from those held by students. Students perceived dishonest behaviors to be more widespread while faculty's estimates were on average more conservative and closer to actual self-reported rates. These results are not consistent with findings from a study conducted in the United States. The reasons for this discrepancy may reflect institutional or cultural factors, which should be explored in future research. PMID- 12090526 TI - Effects of distractor load and temporal target-distractor separation on numerical comparison performance: a stimulus-competition approach. AB - Effects of distractor load and temporal separation between target and distractor onset were studied in a selective attention task. Two experiments required 128 participants to decide as quickly and accurately as possible which of two spatially separated one-digit numerals was larger. Targets were either preceded or succeeded by distractors with intervals of 0, 33, 83, 133, 183, and 233 msec. In Exp. 1, the target digits could appear with 0, 1, 2, or 3 one-digit distractors numerals, representing four levels of distractor load. In Exp. 2, three asterisks were used as distractors. Interference effects of digit distractors varied with distractor load and temporal separation between target and distractor onsets. Moreover, the effects of distractors preceding the target differed from distractors succeeding the target. The pattern of results provided evidence that distractor-interference effects originated from stimulus competition for selection in visual short-term memory. The results were interpreted in terms of attentional engagement theory of Duncan and Humphreys. PMID- 12090527 TI - Response to moral choices as a function of self-esteem. AB - This study investigated the relationship between self-reported self-esteem and academic moral choice by 162 middle school students enrolled in Grades 6, 7, and 8 of public institutions. They were presented nine moral situations (five are school-related) and the Coopersmith Self-esteem Inventory (School Form). Analysis indicated that four of the five school-related moral situations were statistically significantly related to scores of self-esteem. Students reporting higher self-esteem were more likely to make a moral choice in a given academic situation. It is recommended that support of academic honesty can be encouraged by increasing youth self-esteem. PMID- 12090528 TI - Comparison of a multidimensional and a global measure of job insecurity: predicting job attitudes and work behaviors. AB - This paper examined the multidimensional formulation of job insecurity theory described by Greenhalgh and Rosenblatt in 1984. They held that job insecurity is multidimensional and is comprised of two principle threats, (1) threat to the job and (2) threat to job features. To date, there has been no empirical test comparing the predictive value of the separate dimensions of job insecurity on the outcome variables commitment, trust, and job search behavior. A total of 276 managers completed surveys of their job insecurity using both multidimensional and global measures of job insecurity. Regression analysis suggested that the job threat dimension better explains outcome attitudes and behaviors than the job features dimension. Further, contrary to earlier reports, the global measure of job insecurity explained more variance in two out of three outcome variables investigated here. Implications for job insecurity measurement and theory are discussed. PMID- 12090529 TI - Relationship difficulties and unemployment in attempted suicide in an Australian regional center. AB - Over a 12-mo. period, all patients (N = 130) presenting to an Australian regional hospital following attempted suicide were interviewed to investigate the self reported role of unemployment and relationship difficulties in precipitating suicide attempts. Consistent with a study of the same region in 1970-1971, attempted suicide was more prevalent in women, the lower socioeconomic groups, and usually occurred following the threat of or actual separation from a partner. However, in this study attempted suicide was most prevalent for those living in a family situation rather than alone or during separation. Although many patients had made a recent visit to their general practitioner, most acted spontaneously and perceived themselves as having no one with whom they could discuss their feelings or problems. PMID- 12090530 TI - Policy shifts in the presidential debates. AB - To assess whether hearing a debate will be followed by a change in potential voters' ratings of candidates' policy views, 45 undergraduates completed a questionnaire immediately before and after viewing the third presidential debate of 2000 between Bush and Gore. Analysis indicated that agreement with Gore's domestic policy showed a significant shift of 1.2 from pretest to posttest. PMID- 12090531 TI - A critique of Bem's "exotic becomes erotic" theory of sexual orientation development. AB - A critique of Bem's "Exotic Becomes Erotic" theory (1996) on the development of male homosexuality was offered. Limitations were outlined in light of relevant research findings. The authors proposed an alternative model to explain male homosexuality, i.e., reparation of early boyhood trauma, which better accounts for significant psychodynamic elements and available research. Finally, comparisons were made between specific elements of the two models. PMID- 12090532 TI - Reliability of estimates of return of lost letters as a measure of public attitudes to taking human life. AB - An analysis of letters distributed in cities and smaller rural communities in north and south Florida (ns=400 and 448) showed return rates among several addressees were slightly but significantly correlated (rs=.10 and .10) so very small interpoll agreement is present. PMID- 12090533 TI - An evolutionary perspective on the interaction of age and sex differences in short-term sexual strategies. AB - Buss and Schmitt's sexual strategies theory (1993) suggests that short-term mating represents a larger component of men's than women's mating strategies. Assuming this sex difference there is potential for conflict. Symons argued that, because men are more interested in copulation than women, this gives women greater power in establishing conditions (short- vs long-term) under which copulation takes place. The result is that the conflict in sexual strategies is resolved in favor of women's relatively greater interest in long-term sexual strategies. This research tested the hypothesis that across ages men would decrease in desire to employ short-term mating strategies in favor of long-term mating strategies. Specifically, in Study I, men and women in their teens, twenties, and thirties or older were given a measure of desire for a committed relationship. It was predicted that women, regardless of age, would score high on desire for a committed relationship. In contrast, teenage boys would score low on desire for a committed relationship while men in their thirties or older would score as high as the women. In Study II both sexes in their teens, twenties, and thirties or older were given measures of desire for promiscuous sex. It was predicted that the women, regardless of age, would score low on desire for promiscuous sex. In contrast, teenage boys would score high on desire for promiscuous sex while men in their thirties or older would score as low as the women. Support was found for both predictions. PMID- 12090534 TI - Multiscale nonlinearity in a time series of weekly alcohol intake. AB - Several authors have discussed the possibility that nonlinear dynamics might clarify substance abuse and chemical dependence. Nonlinear dynamical structure implies the possibility of oscillatory dynamics and high sensitivity to external perturbation. In this single-subject case study we analyzed the alcohol intake of a substance-abusing participant over a period of about five years using nonlinear time series analysis. The intake is measured in ounces per week and ounces per four weeks--approximately one month--yielding time series at two different time scales. We present statistical evidence that the participant's alcohol intake is nonlinear on both weekly and monthly time scales. We then discuss the implications of this multiscale nonlinearity for our understanding of substance abuse. PMID- 12090536 TI - Interrater agreement on employees' job performance: review and directions. AB - This paper reviews the literature on interrater agreement in 360 degree feedback systems. Multirater performance appraisals, e.g., 360 degree feedback systems, have long been advocated as important mechanisms for enhancing individual and organizational performance. Our review of the empirical literature indicates that interrater agreement is a highly desired but not often accomplished goal in 360 degree feedback systems. A revicw of the assumptions concerning interrater agreement suggests avenues for the evolution of assessment scales used for development and evaluation in organizations. PMID- 12090535 TI - An easy-to-use word processing program for creating concept cards in psychology courses: a method for teachers. AB - We describe a template to create concept cards in psychology courses using a word processing program. Students create their own individualized cards, which have the look and feel of flashcards and retain the same self-testing and monitoring features. Students report the template is easy to use, that the cards help them focus their study behavior and employ critical thinking skills in learning class material. We offer several suggestions on how to use the cards. PMID- 12090538 TI - Moral conflict as a component of ordinary worry. AB - This article contributes to the psychology of moral behavior by inquiring into the presence and extent of worrying about moral concerns in one's life relative to worrying about practical concerns. A 20-item questionnaire was developed, mixing eight moral worry questions with twelve ordinary worry items (finances, health) identified in previous research on worry. Factor analysis produced three domains of worrisome thinking: moral concerns, social desirability, and personal and family health. A single item inquiring into worry about not living up to God's expectations did not load onto any other factor and was dropped from further analysis. Internal consistency for the moral worry factor scale was .85. Mean scores for this scale (moral domain) were significantly lower than mean scores for the two practical worry domain factors. Limitations of the study and directions for further research are outlined. PMID- 12090537 TI - Assessment of attitudes about new learners' roles: factor analysis of the beliefs about Working in Groups Questionnaire. AB - This article presents an analysis of the factor structure of the Beliefs about Working in Groups Questionnaire, which is based on a model of teaching focused on the complementary roles of teachers as models and coaches and students who have to regulate their own learning and learn together with and from peers. This self report questionnaire presents statements describing salient aspects of group work to elicit beliefs students hold about two main aspects of the quality of working in groups, firstly, the belief that working in small groups has important advantages over working individually for developing deep learning; secondly, beliefs that working with peers in close interaction does or does not facilitate learning-focused dialogue. The questionnaire was administered to university sophomores. The hypothesized two-factor structure emerged. It was tested whether the two factors were related to the students' familiarity with working in small groups in high school, to the frequency with which they worked in groups, and to their perception of the value high school teachers attached to working in small groups. PMID- 12090539 TI - Prospects for hepatitis C virus therapeutics: levovirin and viramidine as improved derivatives of ribavirin. AB - Ribavirin, in combination with interferons, has proved clinically useful for the treatment of hepatitis C virus (HCV) infection despite uncertainty as to its true mechanism of action. Its immunomodulatory effects have been proposed to be crucial for its efficacy in HCV treatment. Two new derivatives of zibavirin, Levovirin and virmidine, are currently in development as HCV therapeutics. Both drugs retain ribavirin's immunomodulatory properties but appear to be less toxic than the parent drug. Clinical evaluation of these drugs may aid in understanding the relevant mechanism of action of ribavirin itself, as well as the role of immunomodulators in HCV therapy. PMID- 12090540 TI - XTL-001. XTL Pharmaceuticals. AB - XTL-001, a combination of two human high-affinity monoclonal antibodies with affinity to different regions of the HBV surface antigen (HBsAg), is under development by XTL Biopharmaceuticals as a potential passive immunotherapy for HBV infection. An IND for XTL-001 was submitted in the third quarter of 1998 and was approved by the FDA in March 1999. It has been anticipated that the use of two antibodies against different viral determinants would lower the probability of viral escape mutants. XTL-001 showed viral neutralization activity in the Trimera model for HBV infection and was several thousand-fold more active than current antibody-based therapies. XTL has anticipated that XTL-001 will be used as a combination therapy with existing drugs. In March 2002, Roberton Stephens predicted a 2006 launch for XTL-001, with sales expected to reach at least US$100 million. PMID- 12090541 TI - Interferon-alpha. Amarillo Biosciences. AB - Amarillo Biosciences is developing low-dose oral interferon-alpha (IFNalpha; Veldona) as a potential treatment for primary Sjogren's syndrome, oral mucositis in cancer patients, hepatitis B and C virus (HBV and HCV) infections, and bone marrow disorders. The company is also developing a topical formulation of IFNalpha for the potential treatment of genital warts. The product is registered in Ghana for the treatment of HBV infection. In October 2001, Amarillo licensed rights to its low-dose oral IFNalpha to Atrix Laboratories for the Orphan indications of oral papillomavirus and Behcet's disease. PMID- 12090542 TI - Thymosin alpha1. SciClone Pharmaceuticals. AB - Thymosin alpha1 (Talpha1), a synthetic 28-amino acid peptide with multiple biological activities primarily directed towards immune response enhancement, was originally developed by Alpha 1 Biomedicals for the treatment of hepatitis B virus (HBV) infection. SciClone developed and launched Talpha1, under the trade name Zadaxin, for the treatment of HBV and hepatitis C virus (HCV) infections. The drug is also being developed for the treatment of non-small cell lung cancer (NSCLC), hepatocellular carcinoma, AIDS and malignant melanoma. Talpha1 is able to potentiate the action of cytokines and also reduce the hematological toxicity of cytotoxic drug therapy (cyclophosphamide-, 5-fluorouracil-, dacarbazine- or ifosfamide-based regimens). These studies also demonstrated the mechanism of action of Talpha1 and its role as an immune system enhancer. By July 2001, it was in phase III trials in the US in combination with PEGylated interferon-alpha, and later the same month it was approved in the Philippines. SciClone received expanded approval for HBV and HCV infection in Mexico in July 2001. Talpha1 has been launched in Argentina, China, Peru, the Philippines and Singapore for the treatment of chronic HBV infection. The product subsequently received expanded approval for the treatment of both HBV and HCV infection in Argentina. Marketing approval was granted in India for HBV infection in February 2001. The company was working to expand this approval to include HCV infection. In March 2000, approval for treatment of HBV infection was granted in Thailand, Laos and Malta. Approval was also granted in Sri Lanka and Brunei in August 1999. In September 2000, SciClone announced that approval had been expanded to include the treatment of HCV infection as well as the previously approved HBV indication in both Peru and Sri Lanka. In January 1999, SciClone received approval for Talpha1 in Venezuela for the treatment of HBV and HCV infection. The company also filed a marketing application in New Zealand for Talpha1 to treat HBV infection. The drug was approved in South Korea in April 2000, as an influenza vaccine adjuvant and this was expected to be expanded to indude use for treatment of both HBV and HCV infections. In July 2001, it was approved in In September and October 2000, SciClone was granted patents in Mexico and Canada, respectively, for the use of Talpha1 for the treatment of HCV infection. In June 2000, SciClone was issued a Notice of Allowance by the US Patent and Trademark Office for use of Talpha1 in the treatment of HBV infection. The EPO granted a patent, exclusively licensed to SciClone, for the use of Talpha1 as a monotherapy or in combination with interferon, to treat for HCV infection. In April 2001, SciClone received a Notice of Allowance for a US patent covering newly described analogs of Talpha1. The patent gave the Philippines as an adjuvant to chemotherapy for the treatment of various cancers. In December 2001, Talpha1 entered a phase 1 trial program in Europe, with patient enrolment planned for 2002. SciClone exclusive composition of-matter rights to several families of Talpha1 analogs that could have proprietary therapeutic or biologic distinctions from Talpha1. The company was issued US patents covering the use of Talpha1 for the treatment of HCV infection in August 1998 and the treatment of HBV infection in September 1999. A Notice of Allowance for a second US patent covering the use of Talpha1 was issued in October 1999. In April 1999, SciClone received allowance of a patent from the EPO covering the use of Talpha1 in small cell and non-small cell lung cancer. In August 2001, SciClone received a notice of allowance for patent protection in Japan covering the use of Talpha1. The patent, which extends until 2012, also covers the use of Talpha1 in combination with interferon-alpha for the treatment of HCV infection. SciClone was previously granted a Japanese patent for the use of Talpha1 in the treatment of HBV infection. PMID- 12090543 TI - Immunomodulatory macrolactams for topical treatment of inflammatory skin diseases. AB - Tacrolimus (FK-506; Fujisawa Pharmaceutical Co Ltd) and pimecrolimus (SDZ-ASM 981; Novartis AG) are topical immunomodulators, which provide an alternative to glucocorticosteroids for the topical treatment of atopic dermatitis and other inflammatory dermatoses. Both substances form a complex with cytosolic immunophilins. The complex blocks calcineurin and inhibits the transcription of NF-AT-dependent pro-inflammatory cytokine genes. Multicenter, randomized, double blind clinical trials have shown the efficacy of both substances in atopic dermatitis. We review the physicochemical characteristics, mode of action, pharmacokinetic data, side effects, results of the clincal trials and further indications for tacrolimus and pimecrolimus. PMID- 12090544 TI - Intravenous immunoglobulin: an emerging treatment for immune-mediated skin diseases. AB - Intravenous immunoglobulins (IVIgs) exert a variety of immunomodulating activities and are therefore increasingly being used for the treatment of immune mediated as well as autoimmune diseases. Although the exact mode of action is still poorly understood, the efficacy of IVIg in several diseases, such as thrombocytopenic pupura and Kawasaki disease, has been proven in clinical trials. According to some uncontrolled clinical studies and many case reports there is evidence for a high efficacy of IVIg in the treatment of skin diseases such as autoimmune blistering diseases, lupus erythematosus, dermatomyositis, scleroderma, toxic epidermal necrolysis, atopic dermatitis, chronic autoimmune urticaria and others. In contrast to other commonly used immunomodulating therapeutic strategies, the safety profile of IVIg treatment is significantly better. This update aims to give a current view on the efficacy and safety of IVIg treatment for dermatological diseases. PMID- 12090545 TI - Pimecrolimus. Novartis. AB - Pimecrolimus, an ascomycin macrolactam derivative, is an inhibitor of T-cell and mast-cell activation, developed and launched by Novartis for the potential treatment of psoriasis and allergic, irritant and atopic dermatitis. The topical formulation had been launched in the US by February 2002 for mild-to-moderate atopic dermatitis in patients aged two years and older. At that time, an oral formulation was in development for which launch was anticipated for 2006. In March 2002, pimecrolimus was approved in Denmark, becoming the first non-steroid prescription cream approved for patients from as young as 3 months of age through to adulthood. At this time, Novartis planned to seek approvals in other European countries during 2002 under the Mutual Recognition Procedure, and elsewhere around the globe. In December 2000, Merrill Lynch predicted sales of SFr 100 million in 2002, rising to SFr 330 million in 2004, and in February 2001, the analysts predicted sales of SFr 120 million in 2002, rising to SFr 574 million in 2005. Later in August 2001, Deutsche Bank estimated sales of SFr 150 million in 2002, rising to SFr 550 million in 2005. Following FDA approval in 2002, Morgan Stanley Dean Witter cautiously predicted sales of SFr 60 million rising to SFr 860 million by 2007. PMID- 12090546 TI - IDEC-131. IDEC/Eisai. AB - IDEC, in collaboration with Eisai, is developing IDEC-131 (E6040), a humanized monoclonal antibody (mAb) against CD154, the ligand for CD40 also called CD40L or gp39, for the potential treatment of several autoimmune diseases. IDEC-131 is based on technology that IDEC licensed from Dartmouth Medical School where researchers demonstrated the biological effects of the anti-CD154 antibody in animal models of autoimmunity. In January 2001, phase II trials in psoriasis and idiopathic thrombocytopenic purpura (ITP) were initiated. By january 2002, a phase II trial in Crohn's disease was also ongoing. A pilot, multicenter, multiple-dose phase I trial in moderate-to-severe psoriasis was initiated in January 2001. This trial was ongoing in January 2002. IDEC, in collaboration with Dartmouth Medical School had also initiated a phase I trial in multiple sclerosis by March 1999. IDEC-131 was also previously being developed for systemic lupus ezythematosus (SLE), although no further development for this indication has been reported since the disclosure of disappointing phase II results in April 2000. Analysts at Morgan Stanley predicted in February 2002, that the product would be launched in 2005, with sales of US $25 million, rising to US $75 million in 2006. PMID- 12090547 TI - Genes and engineered cells as drugs for type I and type II diabetes mellitus therapy and prevention. AB - Despite the manageability of diabetes mellitus, complications associated with the disorder necessitate novel approaches to prevent immune-mediated impairment and destruction in type 1 diabetes, as well as the pancreatic insufficiency and peripheral resistance to insulin in type 2 diabetes. Islet transplantation is evolving into a clinical reality to treat type 1 diabetics and novel uses of gene engineering technology promise to result in tolerance to auto-, allo- and xenoantigens as well as microenvironment-specific immunosuppression. Through the use of a variety of gene delivery vehides, an increasing number of studies demonstrate the feasibility of shielding islet transplants and surrogate beta cells from immune rejection by the local secretion of immunosuppressive soluble molecules and anti-apoptotic factors. Although the achievements of gene and cell therapy in type 2 diabetes mellitus are less clear, seminal studies demonstrate the relevance of this approach to the treatment and perhaps prevention of the underlying causes of the disease, including obesity and insulin resistance. In this review, we attempt to illustrate pivotal studies demonstrating the suitability of genes and cells as drugs in type 1 and type 2 diabetes mellitus, and also provide some other targets that may be suitable for clinical utility. PMID- 12090548 TI - Pegvisomant. Pharmacia. AB - Pegvisomant, a polyethylene glycol (PEG) derivative of human growth hormone (GH) that acts as a highly selective GH receptor antagonist, is under development by Pharmacia (formerly Sensus) as a potential treatment for acromegaly. By February 2001, Sensus had submitted an NDA for the treatment of acromegaly, and an approvable letter indicating outstanding issues had been received by July 2001. Pegvisomant was granted Orphan Drug status by the FDA and was designated for Priority Review. Pegvisomant also received Orphan Drug designation in the EU and Japan. In March 2001, additional regulatory filings were being planned for later in 2001. In October 2001, Pharmacia was preparing an NDA in Japan for the treatment of acromegaly. By September 1998, phase 1 trials of the treatment were underway for diabetic retinopathy, and were planned for diabetic nephropathy in 1999. By September 1997, a phase II trial to test the effects of pegvisonant an insulin sensitivity and secretion in type II diabetes patients was underway. However, no development has been reported for these indications since the dates given. By 1994, Sensus had licensed technology for development of GH receptor antagonists from Genentech and Ohio University. Sensus was to pay Genentech, and Genentech was to receive equity in Sensus and royalties from the commercialization of any product resulting from the agreement. In April 2000, the company entered into a licensing agreement with Shearwater Polymers for the PEGylation of pegvisomant using Shearwater's proprietary technology, which is now used to produce the 20-kDa PEG-derivative of pegvisomant. In June 1999, Pharmacia Corp (formerly Pharmacia & Upjohn) signed an agreement to purchase 19.9% of Sensus and to potentially acquire the remainder of the company at a later date. In March 2001, Pharmacia completed its purchase of Sensus. Analysts at Merrill Lynch predicted in February 2002 that the product would be launched in 2003, with US revenues of $20 million, rising to $115 million in 2006. PMID- 12090549 TI - Exubera. Inhale therapeutic systems. AB - Inhale, in colaboration with Pfizer and Aventis Pharma (formerly Hoechst Marion Roussel; HMR), is developing an insulin formulation utilizing its pulmonary delivery technology for macromolecules for the potential treatment of type I and II diabetes. By July 2001, the phase III program had been completed and the companies had begun to assemble data for MAA and NDA filings; however, it was already clear at this time that additional data might be required for filing. By December 2001, it had been decided that the NDA should include an increased level of controlled, long-term pulmonary safety data in diabetic patients and a major study was planned to be completed in 2002, with the NDA filed thereafter (during 2002). US-05997848 was issued to Inhale Therapeutic Systems in December 1999, and corresponds to WO-09524183, filed in February 1995. Equivalent applications have appeared to date in Australia, Brazil, Canada, China, Czech Republic, Europe, Finland, Hungary, Japan, Norway, New Zealand, Poland and South Africa. This family of applications is specific to pulmonary delivery of insulin. In February 1999, Lehman Brothers gave this inhaled insulin a 60% probability of reaching market, with a possible launch date of 2001. The analysts estimated peak sales at $3 billion in 2011. In May 2000, Aventis predicted that estimated peak sales would be in excess of $1 billion. In February 2000, Merrill Lynch expected product launch in 2002 and predicted that it would be a multibillion-dollar product. Analysts Merril Lynch predicted, in September and November 2000, that the product would be launched by 2002, with sales in that year of e75 million, rising to euro 500 million in 2004. In April 2001, Merrill Lynch predicted that filing for this drug would occur in 2001. Following the report of the potential delay in regulatory filing, issued in July 2001, Deutsche Banc Alex Brown predicted a filing would take place in the fourth quarter of 2002 and launch would take place in the first quarter of 2003. In August 2001, Lehman Brothers predicted that launch would take place in the first half of 2002 and that the product would make sales of $475 million in 2003, rising to $875 million in 2004. In the same month, Deutsche Bank predicted that there would be worldwide sales of $50 million in 2003, rising to $400 million in 2005. At this time, analysts at Credit Suisse predicted a launch of the product in 2003, with sales of $70 million in that year, rising to $550 million in 2005. By October 2001, Deutsche Bank predicted sales of $50 million in 2004 and $250 million in 2005. In September 2001, Morgan Stanley predicted sales of $500 million in 2002, rising to $1250 million in 2006. PMID- 12090550 TI - Herbal medicines for neurological diseases. PMID- 12090552 TI - Neural stem cells and neurodegeneration. AB - Neurodegenerative diseases, such as Parkinson's disease, are characterized by a continuous loss of specific populations of neurons. Possible regenerative interventions include transplanting developing neural tissue or neural stem cells into the host brain, and inducing proliferation of endogenous stem cells by pharmacological manipulations. Neural stem cells (NSC), with the capacity to self renew and produce the major cell types of the brain, exist in the developing and adult central nervous system (CNS). These cells can be grown in vitro while retaining the potential to differentiate into nervous tissue. This review focuses on regenerative therapy in neurodegenerative diseases using NSC. PMID- 12090551 TI - Estrogen and Alzheimer's disease. AB - The potential of estrogen as a therapeutic intervention in the prevention or treatment of neurodegenerative diseases has been the object of much research in the past five years. While clinical research has produced highly diverse results in different studies, basic research has uncovered a number ofmolecular mechanisms which dearly label estrogen as a neuroprotective agent. This short review summarizes the various research results that indicate how estrogen can be beneficial to neurons in Alzheimer's disease. PMID- 12090553 TI - Proteopathy: the next therapeutic frontier? AB - The abnormal conformation and assembly of proteins is a probable cause of many degenerative diseases of old age. These proteopathies include such clinically disparate neurological disorders as Alzheimer's disease. Parkinson's disease and Creutzfeldt-Jakob disease, as well as a variety of non-neurological maladies. The involvement of protein pathology in these diseases is well established and we are beginning to understand the process whereby proteins self-assemble and injure tissues; however, we remain largely in the dark regarding the fundamental origins of the proteopathies. Our present knowledge suggests three broad therapeutic approaches to abrogating the proteopathic cascade: reduce the production of the offending proteins, prevent their self-assembly, or promote their removal. PMID- 12090554 TI - Nefiracetam. Daiichi Seiyaku. AB - Nefiracetam (DM-9384) is a cerebral metabolic product under development by Daiichi for the potential treatment of the after-effects of cerebrovascular disorders. By September 1999, nefiracetam was In phase II trials in the US for the treatment of mental symptoms associated with sequelae of stroke and Alzheimer's type dementia. By August 2000, clinical trials in China had been completed in preparation for an NDA filing In November 2000, the companypredicted a launch in 2002. Modified phase III trials of the compound were being repeated in Japan in preparation for re-filing of an NDA. The protocol for the initial Japanese phase III retrial in patients with sequelae of cerebral infarction was approved at the end of July 1999, with completion of the trial expected by the end of 2000. However, in February 2002 Daiichi withdrew its Japanese NDA due to insufficient efficacy in the revised trial. In December 2001, analysts at Morgan Stanley predicted that nefiracetam would be submitted for approval in Japan in 2002, with its approval/launch taking place in late 2002 or 2003. Estimated sales in Japan of yen 3 billon in 2002, rising to yen 13 billion in 2005 and peak sales of yen 20 to 30 billion are predicted. PMID- 12090555 TI - Rasagiline. Teva Pharmaceutical. AB - Rasagiline is a selective and potent irreversible MAO(B) inhibitor which is under development by Teva for the treatment of neurological diseases. Rasagiline is in phase III trials in the US, Canada and Europe for Parkinson's disease (PD) and has completed phase II trials in Israel and Hungary. Teva planned to submit a filing in 2002 and expected to launch rasagiline in 2003. Lundbeck acquired European development and commercialization rights to rasagiline in November 1999 and, in September 2001, the company reported that it planned to file an NDA in 2003. In March 2002, analysts at Morgan Stanley Dean Witter predicted that H Lundbeck would make sales of rasagiline of DKr 100 million in 2003, rising to DKr 300 million in 2008. In the same month, launch was predicted in 2004/2005 for the PD indication, and 2005/2006 for the AD indication, by analysts at Deutsche Banc Alex Brown. PMID- 12090556 TI - Memantine. Merz. AB - Memantine hydrochloride, an NMDA antagonist, was launched in Germany by Merz in 1989 for the treatment of dementia, an indication for which development was continuing in other markets. It is also under development by Merz, Lundbeck, Neurobiological Technologies Inc (NTI) Forest Laboratories and Suntory for the potential treatment of Alzheimer's disease (AD), AIDS-related dementia and pain in patients with neuropathy, and by Allergan for the potential treatment of ocular disease. By July 2001, a regulatory filing for neuropathic pain was expected in 2003. In February 2002, the CPMP recommended to the EU commission to approve memantine for the treatment of moderately severe-to-severe Alzheimer's disease. At this time, marketing authorization was expected late in the first half of 2002, and Lundbeck planned to launch memantine under the brand name Ebixa during the second half of 2002. Merz and Lundbeck, filed memantine for AD in the EU in September 2000 and an NDA was submitted in November of that year. The compound was in phase H trials in the US for the treatment of AIDS-related dementia and pain by August 1996 and phase III trials for glaucoma and neuroprotection by 1999. Analysts at Merrill Lynch predicted in October 2001 that Allergan would make regulatory filings in the US for memantine in glaucoma and ocular hypertension in 2005, and that Forest Laboratories would file for memantine in the US as a supplement to Alzheimer's disease data in early 2002, and for the treatment of neuropathic pain in 2003. Sales of $25 million in 2004, rising to $75 million in 2005, were predicted by Merrill Lynch for this product. PMID- 12090557 TI - Interactions between ionizing radiation and drugs in head and neck cancer: how can we maximize the therapeutic index? PMID- 12090558 TI - Reversers of the multidrug resistance transporter P-glycoprotein. AB - Multidrug resistance can arise from the presence of the membrane-bound pump, P glycoprotein, in a tumor. Major efforts have been made to develop inhibitors of this pump, and a number of promising blockers have reached late stages of clinical trials. The kinetics of the inhibition of P-glycoprotein is complex, with binding sites that can interact synergistically. Reversers of increased affinity and specificity could, in principle, be developed on the basis of these synergies, and offer some promise in cancer therapeutics. PMID- 12090559 TI - Biricodar. Vertex Pharmaceuticals. AB - Vertex is developing biricodar as a chemosensitizing agent designed to restore the effectiveness of chemotherapeutic agents in tumor multidrug resistance. By November 1998, phase II trials had commenced for biricodar, in combination with chemotherapy, for five common cancer indications: breast, ovarian, soft-tissue sarcomas, small cell lung cancer and prostate cancer. Phase II trials were ongoing in January 2002. By March 2000, Vertex was the sole developer of biricodar, as an agreement made in 1996 with BioChem Pharma (now Shire Pharmaceuticals), for the development and marketing of biricodar in Canada was terminated. Biricodar is the free base compound, which also has a citrate salt analog known as VX-710-3. Vertex has published three patents, WO-09615101, WO 09636630 and WO-09736869, disclosing derivatives of biricodar that are claimed for the treatment of multidrug resistant protein and P-glycoprotein-mediated multidrug resistant tumors. In January 2002, a Banc of America analyst report forecast that biricodar had a 30% chance of reaching the market with a launch date in the second half of 2005, with peak sales estimated at $250 million. PMID- 12090560 TI - Revisiting the anticardiolipin test and its standardization. AB - Although the importance of the anticardiolipin test in diagnosis of antiphospholipid syndrome (APS) is widely accepted, there remains much misunderstanding about the strengths and weaknesses of this assay. Several disorders result in formation of low levels of the antibody, hence the anticardiolipin test is not specific when results are low positive. In general, the higher the anticardiolipin level the greater the likelihood of a diagnosis of APS. Hence there have been numerous efforts to enable reproducible measurement of anticardiolipin levels. Standard calibrators were introduced to construct calibration curves from which levels of unknown samples can be derived. Those standard calibrators were made by mixing varying quantities of high positive with normal sera. More recently, calibrators derived from monoclonal anticardiolipin antibodies have been introduced. There are advantages and disadvantages with both types of calibrators. Determination of a precise and reproducible anticardiolipin level is difficult, whatever the calibrators used, because the assay is dependent on several variable components, any of which may fail on any given day. Utilization of a semi-quantitative measure (low, medium, high) may suffice in most clinical settings and would be less subject to error. Validated ELISA kits may offer greater reproducibility, since there is less variability than bench assays set up in very different laboratories. Whether using a kit or a bench assay, meticulous attention to detail offers the best opportunity for precision and reproducibility. PMID- 12090561 TI - Expansion of marginal zone B cells is not sufficient for the development of renal disease in NZBxNZW F1 mice. AB - The mechanisms which govern the production of autoantibodies and of tissue damage during systemic lupus (SLE) are still unclear. In the NZBxNZW F1 (BW) model of SLE glomerulonephritis, the activation and commitment of B cells are thought to play a major role in disease progression. Previous analysis has suggested that these mice have a substantial increase of marginal zone (MZ) B cells before the occurrence of the disease. Owing to the probable role of this B cell subset in autoantibody production, it was important to define the possible link between this abnormality and the occurrence of kidney damage. Using cytofluorometry analysis, we followed the splenic MZ B cell phenotype in different series of mice with shared autoimmune genetic background and histologically defined renal status. By comparing BW females and BW males, NZB and NZW mice, we confirm that BW mice have an increase in MZ B cells but this MZ B cells expansion is not directly linked to tissue lesions. Genetically modified BW female mice with a restricted repertoire of B and T cell antigen receptors, and which do not develop nephritis, exhibit the same increase of MZ B cells, suggesting that this increase does not depend on a specific set of antigens. Moreover, our analysis brings to light a pre-disease state in BW males, with autoantibody production and mesangial deposits. PMID- 12090562 TI - Clinical and immunogenetic characterization of Mexican patients with 'rhupus'. AB - The coexistence of systemic lupus erythematosus and rheumatoid arthritis (rhupus), is a rare clinical condition. To date, 50 cases of rhupus have been described worldwide; however, the lack of clinical criteria for this rheumatic condition has created confusion in the characterization of this disorder. Nevertheless, in this paper we describe a comprehensive clinical and serological characterization of a cohort of 22 Mexican patients with rhupus, supported by generic HLA-DR phenotyping. We found that rhupus patients have a special clinical behavior. In this setting, the signs and symptoms of rheumatoid arthritis prevail, little organic damage associated with systemic lupus erythematosus (SLE) exists and none of the cases present thrombosis or morbidity during pregnancy in spite of presenting a high frequency of anticardiolipin antibodies. We also found an increased frequency of HLA-DR1 and HLA-DR2 alleles compared to healthy ethnically matched controls, systemic lupus erythematosus and rheumatoid arthritis patients. PMID- 12090563 TI - SLE/myositis overlap: are the manifestations of SLE different in overlap disease? AB - Myositis is a rare but recognized complication of systemic lupus erythematosus (SLE). This study compares clinical and laboratory features in patients with SLE complicated by myositis with patients with SLE who do not have myositis. Thus we reviewed the notes of 10 patients with an overlap of biopsy-proven myositis and SLE and compared their clinical, serological and immunogenetic features with 290 patients with SLE without myositis. Our data suggests that patients with SLE associated with myositis are more likely to have alopecia, oral ulcers, erosive joint disease and pulmonary disease but less likely to have renal disease. Our SLE/myositis patients were likely to die at a younger age. The overall disease process seems to be influenced by the presence of anti-RNP autoantibodies. PMID- 12090565 TI - Pregnancy outcome in patients with undifferentiated connective tissue disease: a preliminary study on 25 pregnancies. AB - Undifferentiated connective tissue disease (UCTD) is a group of systemic autoimmune conditions not fulfilling the classification criteria for a definite connective tissue disease (CTD). While an average of 20% of UCTD patients develop a defined CTD during follow-up, the remaining patients maintain an undefined disease. Since pregnancy is considered to be an important factor that may alter the course of autoimmune diseases, we examined 25 pregnancies in 20 UCTD patients being followed at our unit in order to evaluate: (i) the pregnancy outcome; (ii) whether pregnancy is associated with flares of disease activity; and (iii) whether pregnancy may be a trigger for the development of a defined CTD. Twenty two pregnancies (88%) were successfully brought to term, while the remaining three (12%) ended in an abortion in the first trimester. Obstetric complications were observed in six out of the 22 successful pregnancies (27%). Six patients (24%) experienced a disease flare during pregnancy or puerperium, one of whom presented a major flare and developed systemic lupus erythematosus. In the other five patients the manifestations at flare were mild and included arthritis, fever and skin rash. The incidence of flares in a control population of non-pregnant UCTD patients over a period of 1 year was 7%. Although UCTD is a mild condition, the risk of flares during pregnancy appears increased and therefore careful monitoring is as necessary as in other CTD patients. Further prospective studies will be necessary to confirm these preliminary observations. PMID- 12090564 TI - Engagement of complement receptor 2 on the surface of B cells from patients with systemic lupus erythematosus contributes to the increased responsiveness to antigen stimulation. AB - B cells from patients with systemic lupus erythematosus (SLE) display increased responses following cross-linking of the surface antigen receptor. We explored the possibility that the increased responses are at least partially due to simultaneous cross-linking of the complement receptor 2 (CR2). To this end, we stimulated fresh B cells from SLE patients with an anti-IgD antibody conjugated to the Epstein-Barr virus gp350 protein, which binds to CR2, and recorded the free intracytoplasmic calcium response during the first 10 min. Despite the fact that SLE B cells were found to express half as many surface CR2 as normal B cells, both peak responses and the percentage of responding cells were significantly increased in the former. These observations suggest that regulatory molecules such as CR2 are involved in the increased B cell responses in SLE patients. We propose that certain immune complexes that circulate in the sera of SLE patients that have anti-surface immunoglobulin specificities and are decorated with natural ligands of CR2, such as C3d, elicit and promote B cell overactivity. PMID- 12090566 TI - Cardiovascular autonomic function in systemic lupus erythematosus. AB - AIM: To determine whether cardiovascular autonomic function is impaired in systemic lupus erythematosus (SLE). METHODS: A case-control study of 23 patients with SLE was performed. Autonomic symptoms were assessed using a standard questionnaire. Cardiovascular autonomic function was measured using 10 non invasive investigations. There were significant differences between patients and controls in three out of 24 parameters measured during the different tests (P < 0.002). These were reduction in systolic blood pressure at 5 min on head-up tilt, and heart rate responses to isometric exercise and cutaneous cold. Eleven out of 23 patients had an abnormal heart rate, blood pressure or Valsalva response (value below the age corrected 5th centile) while testing compared with six of the controls. Plasma adrenaline and noradrenaline levels were significantly lower in patients vs controls in both the supine (P < 0.05) and tilt position (P < 0.01). Twenty-one of the 23 patients had one or more symptoms that may be attributable to abnormalities in autonomic function. There was no significant association between the number of symptoms and presence of autonomic dysfunction. Cardiovascular autonomic impairment may be demonstrated in some patients with SLE. Symptoms attributable to autonomic dysfunction are common in SLE and autonomic assessment may be required. PMID- 12090567 TI - Oculomotor palsy in six patients with systemic lupus erythematosus. A possible role of antiphospholipid syndrome. AB - The objective of this study was to describe clinical, laboratory, and radiological features in systemic lupus erythematosus (SLE) patients with oculomotor palsy (OMP). Among a cohort of 750 SLE patients receiving follow-up at our unit between 1985 and 2000, all patients with OMP were studied. Clinical and laboratory data were recorded, as well as cerebral magnetic resonance imaging (MRI) findings where available. Immunological tests included tests for presence and specificity of antinuclear and antiphospholipid antibodies. Six patients had OMP, which occurred within the first 3 years in two patients and after more than 20 years in two patients. Four patients had focal neuropsychiatric SLE (NPSLE) manifestations and two had diffuse neurological involvement. The four patients with focal NPSLE were either anticardiolipin- or lupus anticoagulant-positive. Cerebral fluid was abnormal in two of four patients who had this test. In four of the six patients, cerebral MRI showed evidence of vasculopathy. The therapeutic regimens varied, although all six patients initially received high-dose corticosteroids. OMP resolved or improved significantly in all six patients. OMP in SLE occurs in a broad spectrum of clinical situations but is an infrequent manifestation of NPSLE. This case series, together with literature review data, suggests various pathophysiological mechanisms in patients with focal or diffuse neurological symptoms. In patients with focal NPSLE, a possible cause of OMP is microthrombosis associated with presence of antiphospholipid antibodies. PMID- 12090569 TI - Dysphagia in a patient with lupus and review of the literature. AB - Dysphagia is not infrequent in patients with connective tissue diseases such as scleroderma, polymyositis or systemic lupus erythematosus (SLE). It is usually the result of gastro-oesophageal reflux but dysmotility can equally be responsible. A case of dysphagia is described in a patient with SLE, who had developed a rare variety of bullous mucous disease affecting the whole length of oesophagus with spontaneous extrusion of an oesophageal cast. Histological features were suggestive of a variant of rare form of bullous disease in SLE called epidermolysis bullosa acquisita (EBA). This rare association of SLE and EBA involving the oesophagus has not been described in the literature. PMID- 12090568 TI - Dysregulation of the granulocyte-macrophage colony-stimulating factor receptor is one of the causes of defective expression of CD80 antigen in systemic lupus erythematosus. AB - CD80 and CD86, expressed on the antigen-presenting cells (APCs) provide costimulatory signals for T lymphocytes. Recently, defective expression of CD80 has been reported in systemic lupus erythematosus (SLE) although its mechanism is unclear. Here, expression of the B7 antigens induced by interferon-gamma, interleukin-4 or granulocyte-macrophage stimulating-factor (GM-CSF) along the differentiation process of APCs was investigated. In contrast to CD86, expression of CD80 on the CD14+ cells induced by GM-CSF was reduced in SLE. GM-CSF receptor (GM-CSFR) was down-regulated by GM-CSF or phorbol 12-myristate 13-acetate in both of the normal controls and SLE patients, while this change was more remarkable in the latter. In the presence of 1-(5-isoquinolinsulfonyl)-2-methylpiperazine, an inhibitor of protein kinase C, the PMA-induced down-regulation of GM-CSFR was reversed in the normal controls but not in SLE. These data suggest that dysregulation of the GM-CSFR might be associated with the defective expression of CD80, leading to dysfunction of the APCs in SLE. PMID- 12090570 TI - Nodular invasive tracheobronchitis due to Aspergillus in a patient with systemic lupus erythematosus. AB - Nodular or pseudomembranous tracheobronchitis due to infection by Aspergillus species is an uncommon presentation of invasive aspergillosis. Most cases have been described in severely immunocompromised hosts. We describe the case of a 23 year-old woman, with recently diagnosed systemic lupus erythematosus, who developed worsening respiratory function. Bronchoscopy revealed rapid development and progression of multiple nodular plaques in her trachea and bronchi. Endobronchial biopsy demonstrated invasive fungal infection with tissue necrosis and the presence of hyphal elements consistent with aspergillosis. To the best of our knowledge, this is only the second report of fulminant invasive tracheobronchitis due to Aspergillus in a patient with an autoimmune disease. PMID- 12090571 TI - SLE in three ethnic groups XIII. the 'weighted' criteria as predictors of damage. PMID- 12090572 TI - Syndrome X and Hughes syndrome. PMID- 12090573 TI - Quantitative evaluation of abdominal wall perfusion after different types of laparotomy closure using laser-fluorescence videography. AB - Laparotomy closure relies on the incisional edges as anchor for the suture material. The results of these techniques are not satisfying, with failure rates of up to 20%. To investigate the effect of different conventional closure techniques and a novel "bridging technique" on abdominal wall perfusion an animal study was performed in rabbits. Abdominal wall perfusion was measured using the method of dynamic laser-fluorescence videography in the first 72 h of incisional wound healing in 25 animals. Suture tension was controlled with a water-filled polyurethane balloon connected to a pressure detector. The effect of laparotomy closure on abdominal wall tissue perfusion depends significantly on the applied technique and suture tension. Avoiding direct sutures in the incisional edges during laparotomy closure leads to a better tissue perfusion of the incisional region than conventional suture techniques. Suture tension can be controlled and adjusted using a water-filled polyurethane balloon as a pressure sensor. PMID- 12090574 TI - Optimal teaching environment for laparoscopic ventral herniorrhaphy. AB - The introduction of laparoscopic techniques after residency training has created a new paradigm dependent on laparoscopic workshops. This study tested the benefit of an animate course and evaluated the role of proctoring in learning to perform laparoscopic ventral hernia repair (LVHR). Surgeons who had taken a 1-day LVHR course (n = 59) were polled to determine previous experience with laparoscopic procedures and experience with LVHR after the course. Forty-eight (81%) surgeons completing the course responded. Thirty-two (67%) surgeons had performed 179 LVHRS (mean 5.6) since the course. There were no statistically significant differences between the groups performing and not performing LVHR regarding academic/private practice (P=0.8) or opportunities to perform a ventral herniorrhaphy (P = 0.6). Fifteen (31%) surgeons were precepted in their hospital operating room by the lead author. Thirteen (87%) of precepted surgeons had performed a LVHR compared with 19 (58%) of the 33 surgeons taking the course without a precepted intervention (P = 0.05). Surgeons with experience performing laparoscopic inguinal hernia repair, Nissen fundoplication, and common bile duct exploration were more likely to perform LVHR (P=0.0001). Surgeons performing only laparoscopic cholecystectomy tended to be less likely to perform LVHR, nearing statistical significance (P=0.08). Surgeons with prior advanced laparoscopic surgery experience are thus more likely to perform LVHR after participating in a 1-day course. Surgeons precepted in their hospital operating room were also more likely to perform LVHR. Participation in an animate laboratory and a precepted experience can impact the future performance of advanced laparoscopic surgery. PMID- 12090575 TI - Laparoscopic versus open groin hernia repair: meta-analysis of randomised trials based on individual patient data. AB - The EU Hernia Trialists Collaboration was established to provide reliable evaluation of newer methods of groin hernia repair. It involved 70 investigators in 20 countries. The aim was to perform systematic reviews and enhance the value of individual studies by reanalysis of data from randomised controlled trials in meta-analyses. Forty-one eligible trials of laparoscopic versus open groin hernia repair were identified involving 7161 participants (with individual patient data available for 4165). Meta-analysis was performed, using individual patient data where possible. Operation times for laparoscopic repair were longer and there was a higher risk of rare serious complications. Return to usual activities was faster, and there was less persisting pain and numbness. Hernia recurrence was less common than after open non-mesh repair but not different to open mesh methods. The review showed that laparoscopic repair takes longer and has more serious complications, but recovery is quicker with less persisting pain. Reduced hernia recurrence was related to the use of mesh rather than the method of mesh placement. PMID- 12090576 TI - Day surgery for laparoscopic repair of abdominal wall hernias. Our experience in 300 patients. AB - Laparoscopic repair of abdominal wall hernias is still a controversial and nongeneralized therapeutic option. The aim of this paper is to evaluate the results of laparoscopic surgery on abdominal wall hernias at a day-surgery unit and to describe our procedure protocol. Prospective analysis of 300 patients undergoing laparoscopic surgery for abdominal wall hernias was conducted: 260 preperitoneal and 40 intraperitoneal. The patients' clinical features, hernia type, intraoperative and postoperative complications, and follow-up are studied for both types of surgery. All the patients receiving surgery with extraperitoneal laparoscopy were completed as a day-surgical procedure with a rate of conversion to open surgery of 2.3%. Twelve (30%) of the 40 patients operated on for ventral hernias using intraperitoneal laparoscopy required hospitalization: five for perioperative complications and seven for pain (16%). There was no case of infection or mesh rejection. The recurrence rates were 0.78% (two cases) for the inguinal hernias and 2.5% (one case) for the ventral hernias. In conclusion, laparoscopic repair of abdominal wall hernias in a day-surgery setting is an efficient alternative to open surgery. PMID- 12090577 TI - Do absorbable mesh sutures cause less chronic pain than nonabsorbable sutures after Lichtenstein inguinal herniorraphy? AB - Chronic inguinal pain occurs in 20-30% of patients after hernia surgery. A prospective randomized study of 162 patients was undertaken to determine whether absorbable sutures cause less pain than continuous polypropylene fixation of the mesh in the Lichtenstein operation. All patients were operated on under local anesthesia by the same senior surgeon. One-half of the meshes were fixed with continuous 2-0 Dexon II sutures and one-half with continuous 2-0 Prolene sutures. The incidence of pain and recurrence was investigated after a mean follow-up of 2 years. Patient characteristics and operative outcome were similar in the two groups and statistically nonsignificant in both. The rate of significant wound hematomas (n = 3), infections (n = 1), and recurrences (n = 2) were low and not related to the type of sutures. In both study groups 24-26% of the patients felt some pain in follow-up, but over 90% were very satisfied with the operation. The absorbable suture material does not appear to cause less neuropathic pain after Lichtenstein operation than nonabsorbable sutures. PMID- 12090578 TI - Comparison of local and general anesthesia in tension-free (Lichtenstein) hernioplasty: a prospective randomized trial. AB - To compare pulmonary effects, postoperative pain and fatigue, morbidity, patient satisfaction, and cost of different anesthetic techniques for inguinal hernia repair, 50 patients were randomized to local and general anesthesia groups (LA and GA). All patients received the same premedications and the same postoperative analgesic regimen. The standardized postoperative analgesic, intramuscular pyroxicam 20 mg, was given to all patients in the recovery room and an additional 20 mg on the same day was given as requested by each patient. Pulmonary function studies and arterial blood gas analysis were performed 1 h prior to the operation and at the postoperative 8th and 24th hours. All patients underwent Lichtenstein's tension-free hernioplasty. Postoperative pain and fatigue were registered 8 h and 24 h after the operation. A questionnaire was filled out by the patients, and they were asked to give grades for the general comfort of the anesthesia and the surgical procedure (1 = worst, 10 = best). Postoperative pulmonary function tests were significantly poorer in the GA group both on 8th- and 24th-hour measurements (P < 0.05). Patients who underwent LA had significantly lower PCO2 and higher PO2 at the postoperative 8th hour (P<0.05). Mean postoperative pain and fatigue scores revealed a significant difference in favor of local anesthesia at only the 8th hour (P<0.05). There were two complications, one in each group (a hematoma in LA and a urinary retention in GA). Patient satisfaction grades were not different in the two groups. We conclude that LA in inguinal hernia repair does not adversely affect pulmonary functions, patients feel less pain, and patient satisfaction is comparable to that with GA. PMID- 12090579 TI - Inguinal cord cysts. AB - Inguinal cord cysts are found in less than 1% of inguinal hernia dissections. They are benign masses anatomically distinct from scrotal hydroceles that frequently mimic incarcerations or may have unusual tissue components; therefore it is recommended that they be removed when encountered to prevent future symptoms. Five cases of male inguinal cord cysts treated by the same surgeon are reviewed. PMID- 12090580 TI - Handlebar hernia: ultrasonography-aided diagnosis. AB - Traumatic hernia resulting from blunt impalement of the abdominal wall, known as "handlebar hernia," is seldom addressed in the surgical literature, with only 28 previously reported cases. We describe our experience with this rare traumatic hernia diagnosed by physical examination and confirmed by ultrasonography. Published reports suggest handlebar hernia's potential for serious underlying injury and the diagnostic importance of computed tomographic scanning. The case presented here demonstrates the value of bedside ultrasonography in diagnosis confirmation and surgical planning for this condition. PMID- 12090582 TI - Laparoscopic repair of postoperative perineal hernia. AB - Perineal hernia is an uncommon complication of abdominoperineal resection and pelvic exenteration. We present an alternative not previously described for the surgical repair of this type of hernia: laparoscopic repair of postoperative perineal hernia. PMID- 12090581 TI - Handlebar hernia masquerading as an inguinal haematoma. AB - We report a child who sustained a traumatic hernia of the lower abdominal wall after being thrown forward against the handlebar of his bicycle. This is a rare injury in children, and the clinical features mimic an inguinal haematoma. Suspicion should be raised by the immediate appearance of a mass above the inguinal canal following groin injury, particularly if the swelling then disappears with the patient supine. PMID- 12090583 TI - Obturator hernia repair--a new technique. AB - Obturator hernia is a rare condition with few reports in the world literature. There appears to be no consensus on the ideal approach and repair for such a condition. We report a simple, quick technique via a lower midline incision using an autogenous peritoneal fold. It is ideal for the contaminated case and in settings where mesh is not readily available. PMID- 12090584 TI - Anatomoclinical research report. PMID- 12090585 TI - Squamous cell carcinoma of the vulva: study of ploidy, HPV, p53, and pRb. AB - Seventy-one cases of invasive squamous cell carcinoma (ISCC) of the vulva were compared with 18 cases of vulvar intraepithelial neoplasia (VIN) and 21 cases of lichen sclerosus. Ploidy was studied by image analysis, HPV-DNA by PCR, and p53 and pRb by immunohistochemistry. Univariate and multivariate statistical analyses were performed. The mean age of the patients with ISCC was 70.6 years; only 8.5% were < 60 (range, 43-89) years. For the 43 patients with follow-up, FIGO surgical stages were I in 16.2%, II in 48.8%, III in 27.9%, and IV in 6.9%. The 5-year survival was 90% for the patients with curative surgery (vulvectomy and lymphadenectomy) and 32% for those with tumors in stages III to IV. Previous history of nonneoplastic epithelial alterations was recorded in 54%. Vascular invasion was detected in 4.3% and perineural invasion in 21.4%. Inguinal lymph node metastases were present in 34.9% of the cases. Fifty-one (72%) ISCCs were aneuploid, HPV-DNA-16 was detected in 7 (12.3%) cases, overexpression of p53 was found in 40 (56%), and pRb expression was negative in 15 (21.4%). Fifteen cases (80%) of VIN were aneuploid, 5 (27.7%) contained HPV-DNA, 11 (61%) were positive for p53, and all immunoreacted for pRb. All lichen sclerosus cases were diploid, did not contain HPV-DNA, failed to stain for p53, and were positive for pRb. Our study confirmed the prognostic value of conventional pathological features: stage, lymph node metastasis, histological grade, and vascular and perineural invasion; all were statistically significant for survival in the univariate analysis. Also, ploidy was significant in patients with stages I and II tumors. The only significant variable in the multivariate analysis was stage. p53 overexpression appears as a late event in vulvar carcinogenesis, but it may occur before tumor invasion. Lack of pRb expression can occur in vulvar neoplasia, but it does not seem to play any role in the initiation or prognosis of vulvar ISCC. PMID- 12090586 TI - Superficial endometriosis of the uterine cervix: a report of 20 cases of a process that may be confused with endocervical glandular dysplasia or adenocarcinoma in situ. AB - Twenty cases of superficial endometriosis of the uterine cervix that occurred in patients from 20 to 51 (mean 37.1) years of age are described. The majority of the cases were seen in consultation and were usually referred because of diagnostic problems; endocervical glandular dysplasia, adenocarcinoma in situ, or rarely invasive adenocarcinoma were a frequent consideration of the contributor. The endometriosis was almost always an incidental microscopic finding. The endometriotic foci were usually confined to the superficial third of the cervical wall, but in one case there was also involvement of the middle third of the cervical wall. Deep cervical endometriosis was not present in any case. The endometriotic glands were typically evenly spaced and were surrounded at least focally by endometriotic stroma in all cases. The endometriotic stromal cells, however, were significantly obscured by inflammatory cells (two cases), inflammatory cells and hemorrhage (two cases), hemorrhage (four cases), and in one case by smooth muscle metaplasia causing initial failure to recognize the stromal component of the process. The presence of mitotic figures in the glandular epithelium contributed to an initial diagnosis of a premalignant or malignant glandular lesion being made or seriously entertained in 10 cases. Awareness that mitotic figures may be conspicuous in endometriosis from women of reproductive age, the usually bland cytologic features of the endometriotic epithelium, and the presence of associated endometrial stromal cells all facilitate establishing the correct diagnosis. PMID- 12090587 TI - Association between the HLA DQB1*0301 gene and human papillomavirus infection in high-grade cervical intraepithelial neoplasia. AB - This study describes the distribution of DQB1genes in Norwegian women treated for high-grade cervical intraepithelial neoplasia (CIN). Formalin-fixed, paraffin embedded tissue sections from 170 biopsy specimens with diagnoses of CIN II (n = 54) or CIN III (n = 116) were DQB1-typed using allele-specific polymerase chain reaction. The follow-up period for cases was 13 to 15 years. The control material comprised blood samples and endocervical brushes from 213 women without CIN. Both cases and controls had previously been human papillomavirus (HPV)-typed. The DQB1*0301 allele was overrepresented among cases compared with controls (odds ratio [OR] = 1.8). Presence of CIN was related to HPV infection, and HPV 16 positivity was significantly associated with the presence of DQB1*0301 (OR 1.8). The DQBI*0301 allele was significantly more prevalent in CIN III than in CIN II cases. The lesions in two women recurred in the follow-up period, one of whom was carrying the DQB1*0301 allele. Women carrying the HLA-DQB1*0301 allele have an increased risk of developing CIN when infected by HPV 16, although there was not an increased frequency of recurrent disease among women carrying this allele. PMID- 12090588 TI - Utility of MIB-1 in evaluating cauterized cervical cone biopsy margins. AB - Interpretation of cervical cone biopsy margins removed by loop electrosurgical excision procedure is often problematic as a result of cautery artifacts. In this study, MIB-1 expression in cauterized tissues was almost identical to that in the adjacent noncauterized epithelium (correlation coefficients 0.97-0.98). Cauterized dysplastic/condylomatous epithelium showed significantly greater expression of MIB-1 than cauterized normal epithelium (p < or = 0.004) at all levels of the epithelium. Cut-off points for distinguishing dysplastic/condylomatous epithelium from normal epithelium were >15%, 30%, 5%, and 1% of cells in the basal, basal one-third, middle one-third, and surface one third of epithelium, respectively. PMID- 12090589 TI - Infrequent occurrence of high-risk human papillomavirus and of p53 mutation in minimal deviation adenocarcinoma of the cervix. AB - To assess the occurrence of high-risk human papillomavirus (HPV) infection and p53 alterations in minimal deviation adenocarcinoma (MDA) of the uterine cervix, paraffin sections were used to investigate the presence of HPVs 16 and 18 and p53 expression and mutation in six cases of MDA. By polymerase chain reaction, only one case was positive for HPV 16 and none was positive for HPV 18. By in situ polymerase chain reaction in the case positive for HPV 16, HPV 16 was detected in a low-grade squamous intraepithelial lesion overlying the MDA but not in the MDA itself. All of the MDAs were negative or only focally positive for p53 by immunohistochemistry. Our polymerase chain reaction DNA sequencing study failed to detect p53 mutation in exons 5 to 8 in those cases focally positive for p53 expression. These results suggest that MDA may be associated only occasionally with the high-risk HPVs or with p53 gene alterations. PMID- 12090590 TI - Focal necrotizing endometritis: a clinicopathologic study of 15 cases. AB - From routine sign-out of endometrial biopsy specimens, a group of 15 endometria were identified that have a characteristic histologic pattern of inflammation that is not included in present classifications of endometritis. All but one of the women were premenopausal, and all presented with abnormal vaginal bleeding. The lesion is characterized by a patchy, focal inflammation, usually composed of lymphocytes with a variable number of neutrophils and rare macrophages centered around endometrial glands and extending into the glandular lumen with disruption and partial or subtotal necrosis of the endometrial glandular epithelium. These foci were widely dispersed, never confluent, and could be overlooked easily. Plasma cells were not found in any of the endometrial specimens despite methyl green pyronine staining of the samples. Based on the histologic characteristics of this process we have designated it focal necrotizing endometritis. The clinical significance, if any, of focal necrotizing endometritis is currently unknown. PMID- 12090591 TI - Menopausal changes in the myometrium: an investigation using a GnRH agonist model. AB - Thirty-four premenopausal women were randomized to receive 3.75 mg of leuprorelin acetate depot or placebo for 8 weeks before hysterectomy. Postoperatively, the myometrium was examined by two independent pathologists and the pathologic features were graded. Computer analysis was used to assess myometrial cellularity and arterial wall structure (on hematoxylin and eosin-stained sections) and vascularity (on sections immunostained for Factor VIII-related antigen). The cellularity of the gonadotrophin-releasing hormone agonist-treated myometrium was higher than the controls with less stromal edema. Focal myometrial hyalinization was present in a minority of cases, all in the gonadotrophin-releasing hormone agonist-treated cases. The arteries in the gonadotrophin-releasing hormone agonist-treated uteri underwent atrophy of the tunica media and had significantly more perivascular fibrosis. The number of vessels per 100 myocytes also was decreased. Hypoestrinism secondary to leuprorelin treatment leads to myocyte atrophy, decreased stromal edema, atrophy of the arcuate arteries, and decreased myometrial vascularity. PMID- 12090592 TI - Proliferating (LMP) mucinous tumors of the ovaries with microinvasion: morphologic assessment of 13 cases. AB - From a consecutive series of 124 proliferating (low malignant potential) mucinous ovarian tumors, 13 FIGO stage Ia tumors exhibited 1 or more microfoci of stromal invasion. The proliferating mucinous tumors with microinvasion were of intestinal (7/88), endocervical-like (4/26), or mixed epithelial differentiation (2/10) and were more likely to show high-grade (10/42) than low-grade (3/82) epithelial proliferative changes. Morphologically, foci of microinvasion were characterized by single or small groups of epithelial cells in the tumor stroma. These cells were accompanied by a sparse lymphocytic infiltrate and stromal desmoplasia in their immediate vicinity and by a greater degree of nuclear anaplasia in the invading cells and in the lining cells of the nearby glands or cysts than was present elsewhere in the tumors. Focal inflammatory and degenerative changes, commonly seen in ovarian mucinous tumors, were discriminated from microinvasive carcinoma by the nature of the inflammatory response, the presence of stromal mucin deposits, and the absence of nuclear anaplasia. Follow-up was available in 10 of 13 cases (mean 7.5 years) and did not identify any adverse clinical consequences for the patients. PMID- 12090593 TI - Allelic imbalance on chromosome 17p13 in borderline (low malignant potential) epithelial ovarian tumors. AB - Borderline epithelial ovarian tumors (BEOTs) possess clinical and pathologic features intermediate between cystadenomas and cystadenocarcinomas. Although the clinical and pathologic characteristics of BEOTs are well described, the molecular aspects are poorly understood. Three regions of loss of heterozygosity (often referred to as allelic imbalance [AI] when identified by polymerase chain reaction) on chromosome 17p13, one of which includes the p53 gene, have been implicated in the development of ovarian and breast cancers. To provide evidence that genes in these regions also may be involved in the development of BEOTs, we undertook a detailed analysis of AI at all three loci in BEOTs from 21 patients. Seventeen of the BEOTs were serous and four were mucinous. Five of 21 tumors (24%) had AI at one or more loci. Four tumors had AI using the D17S695 marker, two of which showed AI only at this locus. In addition, three tumors exhibited AI at the D17S654 locus, one of which showed AI only at this locus. These data suggest that there may be two tumor suppressor genes distal to p53 involved in the development of at least a subset of BEOTs. Peritoneal implants from a subset of serous BEOTs also were evaluated for AI and were found to be concordant with the primary tumor in all cases. Their genetic similarity is consistent with the implantation theory of peritoneal spread of serous BEOTs in these cases. PMID- 12090594 TI - Sclerosing stromal tumors, thecomas, and fibromas of the ovary: an immunohistochemical profile. AB - The immunohistochemistry of 11 sclerosing stromal tumors (SSTs), 11 fibromas, and 5 thecomas was studied to determine criteria for the assessment of 5 densely sclerotic, calcified ovarian tumors of uncertain diagnosis occurring in young women. The results indicate that the staining pattern for alpha glutathione S transferase can be used to distinguish SSTs, fibromas, and thecomas. CD34, by highlighting the vascular pattern and density, can be used to distinguish between SSTs and other tumors in the thecoma-fibroma group. Alpha-inhibin and calretinin mirrored the alpha glutathione S-transferase staining. Vimentin, smooth muscle actin, and muscle specific actin were generally positive, but desmin was negative in all but one tumor. These results suggest that at least four of the five tumors of uncertain diagnosis were SSTs that had undergone end-stage sclerosis and calcification. PMID- 12090595 TI - Progesterone receptor activity in leiomyomatosis peritonealis disseminata. AB - Leiomyomatosis peritonealis disseminata (LPD) is a rare condition that primarily affects women of reproductive age. Immunohistochemical studies were performed in four cases: LPD from a premenopausal woman on oral contraceptives (one case); LPD associated with postpartum massive ectopic decidual reaction (one case); and LPD from a perimenopausal and a postmenopausal woman. Progesterone receptor activity was present in nine of nine cases, and eight of eight cases were strongly positive for vimentin; reactivity for cytokeratin was uniformly negative. Most cases had a pattern of staining typical of smooth muscle tumors with expression of desmin, smooth muscle actin, and muscle-specific actin. Although estrogen receptor was detected in most cases, reactivity was notably absent (one case) or weak (one case) in nodules with a prominent decidual reaction. Expression of CD 34, a marker for which LPD staining characteristics have not been previously reported, varied from absent to weak. Peritoneal nodules from the postmenopausal woman lacked staining for both estrogen receptor and desmin, smooth muscle actin and muscle-specific actin were only focally expressed, whereas staining for CD 34 was focally intense. Uterine myometrium and leiomyomata were positive for progesterone and estrogen receptor, vimentin, desmin, smooth muscle actin, and muscle-specific actin. Cytokeratin expression was absent. CD 34 exhibited weak staining in leiomyomata, but was absent from myometrium. Progesterone receptor appears to be uniformly expressed in LPD nodules from premenopausal and postmenopausal women, a finding supporting the contention that hormones influence the development of LPD in all cases, regardless of menopausal status. PMID- 12090597 TI - Endocervicosis presenting as a painful vaginal mass: a case report. AB - A case of endocervicosis presenting as a painful vaginal mass in a 36-year-old woman is reported. No previous similar cases have been found in the literature. PMID- 12090596 TI - Primary malignant melanoma of the uterine cervix: case report with world literature review. AB - A 63-year-old patient with a malignant melanoma of the uterine cervix is described. Subtle epitheliotropism of the neoplastic cells within the endocervical columnar epithelium suggested melanoma in situ and the possibility of a primary uterine cervical melanoma, despite a negative anti-S-100 protein immunohistochemical stain. An exhaustive clinical workup, and ultimately, complete autopsy failed to reveal any other primary tumor site, and the diagnosis of melanoma was confirmed by histology and immunohistochemistry on the hysterectomy specimen. A world literature review revealed 54 previously reported cases of uterine cervical melanoma of which 43 had been reported as primary uterine cervical melanoma. A true intraepithelial melanocytic component was found in only 14 of those cases, however, and none of those reports illustrated this with the clarity with which it was seen in the endocervical glandular and surface columnar epithelium of the present case. Primary uterine cervical melanoma is usually discovered at an advanced stage and is no longer amenable to curative therapy. Even when this tumor is discovered early, however, the diagnosis may be unnecessarily delayed if the often subtle interaction of the neoplastic cells with the benign cervical squamous or glandular epithelium is not appreciated, or if the possibility of malignant melanoma is not entertained based on other histologic or immunohistologic characteristics of the tumor cells. PMID- 12090599 TI - History of gynecological pathology. VI. Jacob Felix Marchand. PMID- 12090598 TI - Ovarian mucinous cystadenocarcinoma with yolk sac tumor in a 71-year-old woman. AB - An ovarian mucinous cystadenocarcinoma admixed with a component of yolk sac tumor was encountered in a 71-year-old woman, the first such tumor to be reported. The preoperative serum level of alpha-fetoprotein was elevated (55.6 ng/mL). A right ovarian cystic tumor, 8 cm in maximal dimension, was removed and found to have a solid component. Microscopically, the cystic lesion was a mucinous cystadenocarcinoma, whereas the solid portion was typical yolk sac tumor with microcystic, reticular, and labyrinthine patterns, Schiller-Duval bodies, scattered giant cells with bizarre nuclei, and intracellular and extracellular hyaline globules. There was no evidence of other germ cell tumor components. Yolk sac tumor showed positive reactions for alpha-fetoprotein, carcinoembryonic antigen, and alpha-1-antitrypsin but was negative for CA125, CA19-9, and human chorionic gonadotrophin. The patient died of recurrent tumor 6 months postoperatively without response to combination chemotherapy. PMID- 12090600 TI - Work time, work interference with family, and psychological distress. AB - Despite public concern about time pressures experienced by working parents, few scholars have explicitly examined the effects of work time on work-family conflict. The authors developed and tested a model of the predictors of work time and the relationships between time, work interference with family (WIF). and psychological distress. Survey data came from 513 employees in a Fortune 500 company. As predicted, several work and family characteristics were significantly related to work time. In addition, work time was significantly, positively related to WIF, which in turn was significantly, negatively related to distress. The results suggest that work time fully or partially mediates the effects of many work and family characteristics on WIF. PMID- 12090601 TI - Hindsight bias in economic expectations: I knew all along what I want to hear. AB - Hindsight bias in economic expectations was investigated with particular focus on the moderating effects of attitudes. Stronger hindsight bias was expected for subjectively favorable economic developments. Six months before and after the introduction of the euro as the official book currency of the European Monetary Union, participants rated the probabilities of several economic developments. Results show that hindsight bias occurs with economic expectations and that it is moderated by attitudes. Euro supporters showed stronger hindsight bias for positive economic developments than for negative ones, whereas euro opponents showed the opposite pattern. The results support the notion that hindsight bias is a reconstruction bias in which self-serving tendencies can influence the reconstruction selectively for favorable and unfavorable outcomes. PMID- 12090602 TI - Predictors and outcomes of workplace violence and aggression. AB - The authors developed and assessed the psychometric properties of an instrument measuring risk for workplace violence and expanded a model linking (a) risk and experience of violence and aggression from the public and (b) experience of aggression from coworkers to emotional well-being, psychosomatic well-being, affective commitment, and turnover intentions. Using data from 254 employees representing 71 different occupations, the measure demonstrated acceptable within occupation and 1-month test-retest reliability. The data supported the model and showed that public-initiated violence and aggression and coworker-initiated aggression were differentially associated with personal and organizational outcomes. PMID- 12090603 TI - Neutralizing substitutes for leadership theory: leadership effects and common source bias. AB - The purpose of this research was to examine alternative models of substitutes for leadership theory given the general lack of empirical support for the moderating effects postulated by the theory. On this basis, the research posited that the effects of substitutes also could be conceptualized as mediated relations. The research examined moderated and mediated relations for several sets of leader behaviors and substitutes that have been examined in the literature. The research design sampled 49 organizations, with 940 subordinates rating 156 leaders. Results, although generally not supportive of the moderator or mediator hypotheses, essentially demonstrated that leadership matters. The findings also suggest that prior significant effects in substitutes literature may be merely a statistical artifact, resulting from common-source bias. PMID- 12090604 TI - An examination of the antecedents and moderator influences of climate strength. AB - The authors tested 3 hypotheses regarding the antecedents and moderator influences of climate strength (CS; the degree of within-unit agreement of climate perceptions). The sample consisted of 197 work units. Social interaction among unit members showed positive, statistically significant correlations with CS in goals orientation and innovation climate. Work-unit leaders' informing behavior was positively correlated with CS in the 3 climate facets measured (i.e., support, goals orientation, and innovation). CS in innovation moderated the impact of work units' innovation climate on average satisfaction and commitment. CS in goals orientation moderated the influence of work units' goal orientation on average commitment. The moderator influences showed the expected direction: CS fostered the influence of work units' climate on the criterion variables. PMID- 12090605 TI - Commitment to organizational change: extension of a three-component model. AB - Three studies were conducted to test the application of a three-component model of workplace commitment (J. P. Meyer & N. J. Allen, 1991: J. P. Meyer & L. Herscovitch, 2001) in the context of employee commitment to organizational change. Study 1, conducted with 224 university students, provided preliminary evidence for the validity of newly developed Affective, Continuance, and Normative Commitment to Change Scales. Studies 2 and 3, conducted with hospital nurses (N = 157 and 108, respectively), provided further support for the validity of the three Commitment to Change Scales, and demonstrated that (a) commitment to a change is a better predictor of behavioral support for a change than is organizational commitment, (b) affective and normative commitment to a change are associated with higher levels of support than is continuance commitment, and (c) the components of commitment combine to predict behavior. PMID- 12090606 TI - Development and test of a model linking safety-specific transformational leadership and occupational safety. AB - The authors developed, tested, and replicated a model in which safety-specific transformational leadership predicted occupational injuries in 2 separate studies. Data from 174 restaurant workers (M age = 26.75 years, range = 15-64) were analyzed using structural equation modeling (LISREL 8; K. G. Joreskog & D. Sorbom, 1993) and provided strong support for a model whereby safety-specific transformational leadership predicted occupational injuries through the effects of perceived safety climate, safety consciousness, and safety-related events. Study 2 replicated and extended this model with data from 164 young workers from diverse jobs (M age = 19.54 years, range = 14-24). Safety-specific transformational leadership and role overload were related to occupational injuries through the effects of perceived safety climate, safety consciousness, and safety-related events. PMID- 12090607 TI - Goal orientation and ability: interactive effects on self-efficacy, performance, and knowledge. AB - This study examined the direct relationship of goal orientation--and the interaction of goal orientation and cognitive ability--with self-efficacy, performance, and knowledge in a learning context. The authors argue that whether a particular type of goal orientation is adaptive or not adaptive depends on individuals' cognitive ability. Consistent with previous research, learning orientation was positively related to self-efficacy, performance, and knowledge, whereas performance orientation was negatively related to performance only. The interactions between goal orientation and ability also supported several hypotheses. As expected, learning orientation was generally adaptive for high ability individuals but had no effect for low-ability individuals. In contrast, the effects of performance orientation were contingent on both individuals' level of cognitive ability and the outcome examined. PMID- 12090608 TI - Two studies examining the negative effect of self-efficacy on performance. AB - Although hundreds of studies have found a positive relationship between self efficacy and performance, several studies have found a negative relationship when the analysis is done across time (repeated measures) rather than across individuals. W. T. Powers (1991) predicted this negative relationship based on perceptual control theory. Here, 2 studies are presented to (a) confirm the causal role of self-efficacy and (b) substantiate the explanation. In Study 1, self-efficacy was manipulated for 43 of 87 undergraduates on an analytic game. The manipulation was negatively related to performance on the next trial. In Study 2, 104 undergraduates played the analytic game and reported self-efficacy between each game and confidence in the degree to which they had assessed previous feedback. As expected, self-efficacy led to overconfidence and hence increased the likelihood of committing logic errors during the game. PMID- 12090609 TI - Measurement equivalence: a comparison of methods based on confirmatory factor analysis and item response theory. AB - Current interest in the assessment of measurement equivalence emphasizes 2 major methods of analysis. The authors offer a comparison of a linear method (confirmatory factor analysis) and a nonlinear method (differential item and test functioning using item response theory) with an emphasis on their methodological similarities and differences. The 2 approaches test for the equality of true scores (or expected raw scores) across 2 populations when the latent (or factor) score is held constant. Both approaches can provide information about when measurement nonequivalence exists and the extent to which it is a problem. An empirical example is used to illustrate the 2 approaches. PMID- 12090610 TI - Five-factor model of personality and job satisfaction: a meta-analysis. AB - This study reports results of a meta-analysis linking traits from the 5-factor model of personality to overall job satisfaction. Using the model as an organizing framework, 334 correlations from 163 independent samples were classified according to the model. The estimated true score correlations with job satisfaction were -.29 for Neuroticism, .25 for Extraversion, .02 for Openness to Experience, .17 for Agreeableness, and .26 for Conscientiousness. Results further indicated that only the relations of Neuroticism and Extraversion with job satisfaction generalized across studies. As a set, the Big Five traits had a multiple correlation of .41 with job satisfaction, indicating support for the validity of the dispositional source of job satisfaction when traits are organized according to the 5-factor model. PMID- 12090611 TI - Coworker exchange: relationships between coworkers, leader-member exchange, and work attitudes. AB - The study of leadership exchanges is extended by studying both leader-member exchanges (LMXs) and coworker exchanges (CWXs). Data from 110 coworker dyads were used to examine relationships between LMXs and CWXs and between exchange relationships and work attitudes. As predicted, the interaction between 2 coworkers' LMX scores predicted CWX quality for the coworker dyad. Also, after controlling for LMX, greater diversity in a worker's CWX relationships was negatively related to his or her organizational commitment but not job satisfaction. The quality of a worker's CWX relationships, however, did not moderate the relationship between CWX diversity and work attitudes. PMID- 12090612 TI - The role of different levels of leadership in predicting self- and collective efficacy: evidence for discontinuity. AB - This study identified potential discontinuities in the antecedents of efficacy beliefs across levels of analysis, with a particular focus on the role of leadership climate at different organizational levels. Random coefficient modeling analyses conducted on data collected from 2,585 soldiers in 86 combat units confirmed that soldiers' experience, role clarity, and psychological strain predicted self-efficacy to a greater extent than did leadership climate. Also, leadership climate at a higher organizational level related to self-efficacy through role clarity, whereas leadership climate at a lower organizational level related to self-efficacy through psychological strain. Group-level analyses identified leadership climate at a higher organizational level as the strongest predictor of collective efficacy. Theoretical and practical implications and directions for future research are discussed. PMID- 12090614 TI - Perceived supervisor support: contributions to perceived organizational support and employee retention. AB - Three studies investigated the relationships among employees' perception of supervisor support (PSS), perceived organizational support (POS), and employee turnover. Study 1 found, with 314 employees drawn from a variety of organizations, that PSS was positively related to temporal change in POS, suggesting that PSS leads to POS. Study 2 established, with 300 retail sales employees, that the PSS-POS relationship increased with perceived supervisor status in the organization. Study 3 found, with 493 retail sales employees, evidence consistent with the view that POS completely mediated a negative relationship between PSS and employee turnover. These studies suggest that supervisors, to the extent that they are identified with the organization, contribute to POS and, ultimately, to job retention. PMID- 12090615 TI - One role or two? The function of psychological separation in role conflict. AB - Student athletes vary in how much they view their academic and athletic role identities as separate from and interfering with each other. The authors investigated the relation of these perceptions to psychological well-being in 200 intercollegiate athletes. Measures included role separation, interference, identity, and well-being. Correlations indicated that interference related negatively to well-being, whereas viewing the roles as distinct related positively to well-being. Regression analyses of demographic and role identity variables also showed a positive association between role separation and well being, and a significant Separation x Interference interaction. Specifically, role interference was negatively related to well-being for those who viewed the two roles as distinct but unrelated for those who did not. The buffering effects of role separation are discussed. PMID- 12090613 TI - Violence and sex impair memory for television ads. AB - Participants watched a violent, sexually explicit, or neutral TV program that contained 9 ads. Participants recalled the advertised brands. They also identified the advertised brands from slides of supermarket shelves. The next day, participants were telephoned and asked to recall again the advertised brands. Results showed better memory for people who saw the ads during a neutral program than for people who saw the ads during a violent or sexual program both immediately after exposure and 24 hr later. Violence and sex impaired memory for males and females of all ages, regardless of whether they liked programs containing violence and sex. These results suggest that sponsoring violent and sexually explicit TV programs might not be a profitable venture for advertisers. PMID- 12090616 TI - On the bias of Huffcutt and Arthur's (1995) procedure for identifying outliers in the meta-analysis of correlations. AB - This study documents how the use of A. I. Huffcutt & W. A. Arthur's (1995) sample adjusted meta-analytic deviancy (SAMD) statistic for identifying outliers in correlational meta-analyses results in inaccuracies in mean r. Monte Carlo simulations found that use of the SAMD resulted in the overidentification of small relative to large correlations as outliers. Furthermore, this tendency to overidentify small correlations was found to increase as the magnitude of the population correlation increased and resulted in mean rs that overestimated the population correlation. The implications for meta-analysts are discussed, and 2 possible solutions are offered. PMID- 12090617 TI - The role of fair treatment and rewards in perceptions of organizational support and leader-member exchange. AB - This study examined a model of the antecedents and consequences of perceived organizational support (POS) and leader-member exchange (LMX). It was predicted that organizational justice (procedural and distributive justice) and organizational practices that provide recognition to the employee (feelings of inclusion and recognition from upper management) would influence POS. For LMX, it was predicted that leader reward (distributive justice and contingent rewards) and punishment behavior would be important antecedents. Results based on a sample of 211 employee-supervisor dyads indicated that organizational justice, inclusion, and recognition were related to POS and contingent rewards were related to LMX. In terms of consequences, POS was related to employee commitment and organizational citizenship behavior, whereas LMX predicted performance ratings. PMID- 12090618 TI - Structural contingency theory and individual differences: examination of external and internal person-team fit. AB - This article develops and tests a structurally based, integrated theory of person team fit. The theory developed is an extension of structural contingency theory and considers issues of external fit simultaneously with its examination of internal fit at the team level. Results from 80 teams working on an interdependent team task indicate that divisional structures demand high levels of cognitive ability on the part of teammembers. However, the advantages of high cognitive ability in divisional structures are neutralized when there is poor external fit between the structure and the environment. Instead, emotional stability becomes a critical factor among teammembers when a divisional structure is out of alignment with its environment. Individual differences seem to play little or no role in functional structures, regardless of the degree of external fit. PMID- 12090619 TI - Reactive oxygen intermediates and glutathione regulate the expression of cytosolic ascorbate peroxidase during iron-mediated oxidative stress in bean. AB - Excess of free iron is thought to harm plant cells by enhancing the intracellular production of reactive oxygen intermediates (ROI). Cytosolic ascorbate peroxidase (cAPX) is an iron-containing, ROI-detoxifying enzyme induced in response to iron overload or oxidative stress. We studied the expression of cAPX in leaves of de rooted bean plants in response to iron overload. cAPX expression, i.e., mRNA and protein, was rapidly induced in response to iron overload. This induction correlated with the increase in iron content in leaves and occurred in the light as well as in the dark. Reduced glutathione (GSH), which plays an important role in activating the ROI signal transduction pathway as well as in ROI detoxification, was found to enhance the induction of APX mRNA by iron. To determine whether cAPX induction during iron overload was due to an increase in the amount of free iron, which serves as a co-factor for cAPX synthesis, or due to iron-mediated increase in ROI production, we tested the expression of APX in leaves under low oxygen pressure. This treatment, which suppresses the formation of ROI, completely abolished the induction of cAPX mRNA during iron overload, without affecting the rate of iron uptake by plants. Taken together, our results suggest that high intracellular levels of free iron in plants lead to the enhanced production of ROI, which in turn induces the expression of cAPX, possibly using GSH as an intermediate signal. We further show, using cAPX antisense transgenic plants, that cAPX expression is essential to prevent iron mediated tissue damage in tobacco. PMID- 12090620 TI - Specificity of auxin regulation of gibberellin 20-oxidase gene expression in pea pericarp. AB - During early pea fruit growth, the physiological roles of 4-chloroindole-3-acetic acid (4-Cl-IAA) and IAA, natural pea auxins, in regulating gibberellin (GA) 20 oxidase gene expression (PsGA20ox1) were tested with 4-position, ring-substituted auxins that have a range of biological activities (fruit growth). The effect of seeds, and natural and synthetic auxins (4-Cl-IAA, and IAA; 4-Me-IAA, 4-Et-IAA and 4-F-IAA, respectively), and auxin concentration (4-Cl-IAA) on PsGA20ox1 mRNA levels in pea pericarp were investigated over a 24 h treatment period. The ability of the 4-substituted auxins to increase PsGA20ox1 mRNA levels in deseeded pericarp was correlated with their ability to stimulate pericarp growth. The greatest increase in pericarp PsGA20ox1 mRNA levels and growth was observed when deseeded pericarps were treated with the naturally occurring pea auxin, 4-Cl-IAA; however, IAA was not effective. Silver thiosulfate, an ethylene action antagonist, did not reverse IAA's lack of stimulation of PsGA20ox1 over the control treatment. 4-Me-IAA was the second most active auxin in stimulating PsGA20ox1 and was the second most biologically active auxin. Application of the 4 substituted IAA analogs, 4-Et-IAA and 4-F-IAA, to deseeded pericarps resulted in minimal or no increase in PsGA20ox1 transcript levels or pericarp growth. Pericarp PsGA20ox1 mRNA levels increased with increasing 4-Cl-IAA concentration and showed transitory increases at low 4-Cl-IAA treatments (30 to 300 pmol). These results support a unique physiological role for the auxin 4-Cl-IAA in the regulation of GA metabolism by effecting PsGA20ox1 expression during early pea fruit growth. PMID- 12090621 TI - Regulation of carbonic anhydrase gene expression in cotyledons of cotton (Gossypium hirsutum L.) seedlings during post-germinative growth. AB - Recently, plastidial carbonic anhydrase (CA) cDNA clones encoding functional carbonic anhydrase enzymes were isolated from a 48 h dark-grown cotton seedling (cotyledons) cDNA library (Hoang et al., Plant Cell Physiol. 40: 1999). Here we examined the levels of relative transcript abundance and enzyme activities in cotyledons at different developmental stages and under different environmental conditions (i.e. altering CO2 and light conditions), during post-germinative seedling growth. Relative CA transcript levels and total CA enzyme activity in cotyledons of cotton seedlings increased from 18 h to 72 h of post-germinative growth in the dark, although somewhat later than the glyoxylate cycle enzyme, MS. When 24 h old seedlings were exposed to light for an additional 24 h, CA activity in greening cotyledons increased about 2-fold (compared with controls kept in the dark), whereas relative CA transcript levels were essentially the same. Removal of seed coats from cotyledons of 24 h old seedlings dramatically increased relative CA transcript abundance (measured 24 h later) in the dark, but did not influence CA enzyme activity. Manipulation of external CO2 environments (zero, ambient, or high) modulated coordinately the relative transcript abundance of CA (and rbcS) in cotyledons, but did not affect enzyme activity. On the other hand, regardless of the external CO2 conditions, cotyledons of seedlings exposed to light exhibited increased CA activity, concomitant with increased Rubisco activity and increased chlorophyll content. Collectively, our data suggest that steady-state levels of CA and rbcS transcripts are increased in response to environmental CO2 conditions, while CA (and Rubisco) enzyme activities are likely modulated at the post-transcriptional level following exposure of seedlings to light, and in parallel with development of functional chloroplasts. PMID- 12090622 TI - Regulation and characterization of four CBF transcription factors from Brassica napus. AB - Four orthologues of the Arabidopsis CBF/Dreb transcriptional activator genes were isolated from the winter Brassica napus, cv. Jet neuf. All four BNCBF clones encode a putative DRE/CRT (LTRE)-binding protein with an AP2 DNA-binding domain, a putative nuclear localization signal and a possible acidic activation domain. Deduced amino acid sequences suggested that BNCBFs 5, 7and 16 are very similar to the Arabidopsis CBFI whereas BNCBF17 is different in that it contains two extra regions of 16 and 21 amino acids in the acidic domain. Transcripts hybridizing specifically to BNCBF17 and to one or more of the other BNCBFs accumulated in leaves within 30 min of cold exposure of the Brassica seedlings and preceded transcript accumulation of the cold-inducible BN28 gene, a Brassica orthologue of the cor6.6 or KIN gene from Arabidopsis. Cold-induced accumulation of BNCBF17 mRNA was rapid but was short-lived compared to transcripts hybridizing to BNCBF5/7/16. Transcripts hybridizing to one or more of BNCBF5/7/16 accumulated at low levels after the plants were subjected to prolonged exposure to salt stress. BNCBF17 was not responsive to salt stress. BNCBF transcript accumulation was similar in both spring and winter Brassica but the persistence of the transcripts in the cold were generally shorter in the spring than in the winter type. BNCBF5 and 17 proteins bind in vitro to the LTRE domains of the cold-inducible BN115 (cor15a orthologue) or BN28 promoters. Differential binding preferences, however, to LTREs between BNI 15 and BN28 were observed. Mutation of the core CCGAC sequence of the LTRE indicated that BNCBF17 had a lower sequence binding specificity than BNCBF5. Furthermore, experiments indicated that the LTREs were able to drive BNCBF5 and 17 trans-activation of the Lac-Z reporter gene in yeast. We conclude that the BNCBFs reported here could function as trans-acting factors in low-temperature responses in Brassica, controlling the expression of cold induced genes through an ABA-independent pathway. PMID- 12090623 TI - Wounding activates immediate early transcription of genes for ERFs in tobacco plants. AB - We have previously demonstrated that cutting induces the rapid response of genes for ethylene-responsive transcription factors (ERFs) in leaf strips of tobacco, and that the induction was not interfered but enhanced in the presence of the protein synthesis inhibitor cycloheximide (CHX). In this study, we analyzed the expression of genes for ERFs in tobacco plants by injuring leaf tissues with a hemostat. The results verified that mechanical damage is a trigger for rapid and concurrent induction of both the local and the systemic expression of genes for ERFs in tobacco plants. Further studies on systemic response of ERF genes in response to different severity and position of the wound on a leaf suggested that a threshold value might exist for the magnitude of wound signal to induce systemic activation of these genes. Then, we examined expression of genes for ERFs by analysis in transgenic tobacco plants that harbored reporter genes in which the promoter of the gene for NsERF2, NsERF3 or NsERF4 was fused to a gene for beta-glucuronidase. The results suggested that the local and systemic accumulation of ERF mRNAs after wounding was primarily mediated by the rapid activation of transcription of the respective genes. In addition, we found that cycloheximide triggered rapid activation of genes for ERFs which might be mediated via activation of transcription of the genes for ERFs. PMID- 12090624 TI - Regulation of the high-affinity ammonium transporter (BnAMT1;2) in the leaves of Brassica napus by nitrogen status. AB - Substantial concentrations of NH4+ are found in the apoplast of the leaves of Brassica napus. Physiological studies on isolated mesophyll protoplasts with 15NH4+ revealed the presence of a high-affinity ammonium transporter that shared physiological similarity to the high-affinity NH4+ transporters in Arabidopsis thaliana (AtAMT1;3). PCR techniques were used to isolate a full-length clone of a B. napus homologue of AMT1 from shoot mRNA which showed 97% similarity to AtAMT1;3. The full-length cDNA when cloned into the yeast expression vector pFL61 was able to complement a yeast mutant unable to grow on media with NH4+ as the sole nitrogen source. Regulatory studies with detached leaves revealed a stimulation of both NH4+ uptake and expression of mRNA when the leaves were supplied with increasing concentrations of NH4+. Withdrawal of NH4+ supply for up to 96 h had little effect on mRNA expression or NH4+ uptake; however, plants grown continuously at high NH4+ levels exhibited decreased mRNA expression. BnAMT1;2 mRNA expression was highest when NH4+ was supplied directly to the leaf and lowest when either glutamine or glutamate was supplied to the leaves, which directly paralleled chloroplastic glutamine synthetase (GS2) activity in the same leaves. These results provide tentative evidence that BnAMT1;2 may be regulated by similar mechanisms to GS2 in leaves. PMID- 12090625 TI - Potato hexokinase 2 complements transgenic Arabidopsis plants deficient in hexokinase 1 but does not play a key role in tuber carbohydrate metabolism. AB - Potato plants (Solanum tuberosum L. cv. Desiree) transformed with sense and antisense constructs of a cDNA encoding the potato hexokinase 2 exhibited altered enzyme activities and expression of hexokinase 2 mRNA. Measurements of the maximum catalytic activity of hexokinase revealed an 11-fold variation in leaf (from 48% of the wild-type activity in antisense transformants to 446% activity in sense transformants) and an 8-fold variation in developing tubers (from 35% of the wild-type activity in antisense transformants to 212% activity in sense transformants). Despite the wide range of hexokinase activities, no substantial change was found in the fresh weight yield, starch, sugar and metabolite levels of transgenic tubers. However, both potato hexokinases 1 and 2 were able to complement the hyposensitivity of antisense hexokinase 1 Arabidopsis transgenic plants to glucose. In an in vitro bioassay of seed germination in a medium with high glucose levels, double transformants showed the same sensitivity to glucose as that of the wild-type ecotype, displaying a stunted phenotype in hypocotyls, cotyledons and roots. PMID- 12090626 TI - KAP-2, a protein that binds to the H-box in a bean chalcone synthase promoter, is a novel plant transcription factor with sequence identity to the large subunit of human Ku autoantigen. AB - The KAP-2 protein that binds to the H-box (CCTACC) element in the bean CHS15 chalcone synthase promoter was purified, and internal peptide sequence used to design primers leading to the cloning of KAP-2 from bean (Phaseolus vulgaris) and barrel medic (Medicago truncatula). KAP-2 shares sequence similarity to the large subunit of mammalian Ku autoantigen, a protein proposed to be involved in control of DNA recombination and transcription. KAP-2 sequences were present in genomic DNA from a range of legumes, and a related protein is found in Arabidopsis thaliana. Recombinant KAP-2 expressed in insect cells showed the same binding specificity for the CHS15 H-box as the protein purified from bean cell extracts. In vitro transcription assays confirmed that KAP-2 stimulates transcription from a promoter harboring the H-box cis element. PMID- 12090630 TI - Normal values and standard deviations for pupil diameter and interpupillary distance in subjects aged 1 month to 19 years. AB - Normal values of pupil diameters and interpupillary distances (PDs) were measured in a population of 1311 subjects (in 4294 visits) ranging from 1 month of age to slightly over 19 years of age. Subjects in this study were recruited from birth announcements in a local newspaper for a developmental vision project. Pupil sizes were measured photographically when the corneas were illuminated by 15.9 +/ 0.5 lux ambient illumination (i.e. under mesopic conditions). Interpupillary distance was measured with an interocular distance rule while the subject fixated an object at 0.66 m distance. These PD measurements were corrected for systematic measurement errors and to an infinite viewing distance using radii of ocular rotation based on age-dependent axial lengths. Means and S.D. were calculated for age, pupil diameter and PD for each 1-year group of male and female subjects. The second order regression equation for average pupil size as a function of age was determined: [males pupil diameter (in mm) = 5.83 +/- 0.181*age in years - 0053*age in years2, r2 = 0.897; female pupil diameter = 5.40 + 0.285*age in years - 0.0109*age in years2, r2 = 0.945]. The dierence between male and female pupil sizes (mean male - female = 0.13 mm) was marginally not significant (p < 0.054). The average corrected PDs as a function of age were found to approximate another second-order regression equation: (males PD = 43.36 + 1.663*age in years - 0.034*age in years2, r2 = 0.986; females PD = 41.76 + 1.891 *age in years - 0.052*age in years2, r2 = 0.986). Male PD was wider than female PD by an average of 1.58 mm (p < 0.0003). As expected, the results of this study were similar to a preliminary investigation conducted by Thunyalukul et al. [Invest. Ophthalmol. Vis. Sci. 37 (1996) S731] on a portion of the present data set, and also very similar to data from another study of comparable racial composition using a different measurement method [Pryor, Pediatrics 44 (1969) 973]. It was concluded that pupil diameter and PD increase more gradually than axial length of the eye in the first few years of life. The normal values and S.D. for both pupil size and PD determined in this study have important clinical implications as well as applications in the optical industry. PMID- 12090631 TI - Optometrists' examination and referral practices for patients presenting with flashes and floaters. AB - INTRODUCTION: Patients experiencing flashes and floaters commonly present to their optometrist. Some of these patients may have significant pathology, yet there is a great deal of variability with regard to examination technique and referral practice. METHODS: A questionnaire survey was undertaken to determine the current management of patients presenting to their optometrist with flashes and floaters. All practising community optometrists within Southern Derbyshire received a questionnaire and 74 (56.9%) completed replies were received. RESULTS: Optometrists estimated that an average of 14 patients per month per optometrist presented with symptoms of flashes and/or floaters. Mydriasis was utilised routinely for examination in approximately half of the patients. Mean relative confidence was 2.0 at identifying a vitreous haemorrhage and 6.5 for vitreous pigment (complete confidence = 0, complete lack of confidence = 10). Eight percent of responders were unfamiliar with the clinical sign of vitreous pigment, and 17% identifying this sign did not refer all such patients to the hospital services. CONCLUSIONS: Patients presenting to their optometrists with flashes and/or floaters make up a sizeable part of the community optometrist's workload and the management of these patients is highly variable. A large proportion of these patients are examined without mydriasis, even in the presence of various risk factors for retinal detachment. There is a relative lack of confidence amongst optometrists with regards the detection of vitreous pigment and the prognostic implications of this finding. Educational measures such as study days may help the level of understanding and heighten the appreciation of the implications of flashes and floaters and the various clinical signs encountered. PMID- 12090629 TI - Cloning of the promoter for a novel barley gene, Lem1, and its organ-specific promotion of Gfp expression in lemma and palea. AB - The differential display method was used to identify a novel barley gene, Lem1, expressed primarily in the outer organs (lemma and palea) that enclose developing florets and seeds. The promoter was isolated from a BAC genomic clone and used in a translational fusion with a green fluorescent protein gene (Gfp) to produce a transient expression vector. After particle bombardment, Gfp was expressed only in lemmas, paleas and awns of developing spikelets. Lem1 did not promote Gfp expression in vegetative leaves or in mature spikes, although expression of co bombarded uidA (GUS) occurred under the regulation of a ubiquitin promoter. This reproduced the developmentally regulated pattern of mRNA accumulation. Deletion studies showed that the promoter activity is confined to a cis element within 80 bp of the transcription start site. Upstream from this, the promoter contains putative auxin-, ethylene- and gibberellin-responsive elements or homologues. Lem1 was found to be a single intronless gene encoding an acidic 102 amino acid protein, possibly associated with membranes. In a two-rowed barley, Lem1 mRNA was absent in the lateral spikelets, which fail to develop, and present only in the developing median spikelets. This suggests that Lem1 may play a role in organ development. PMID- 12090628 TI - Regulation of CDPKs, including identification of PAL kinase, in biotically stressed cells of French bean. AB - Changes in protein kinase activity have been investigated during the early response of suspension cultured cells of French bean to fungal elicitor. One of the kinases activated has a known target, phenylalanine ammonia-lyase (PAL), which has an important role in plant defence responses, and was purified. Kinase acivity during purification was monitored for both the PAL-derived peptide and syntide-2, which it also phosphorylated. The kinase had an Mr of 55,000 on the basis of gel migration, 45Ca2+ binding, autophosphorylation and phosphorylation of various substrates using in-gel assays. The kinase has been characterised with respect to kinetics and other properties in vitro and appears to be a CDPK. In gel assays were also used to show that this kinase and a number of other CDPKs of similar Mr showed complex changes in elicitor-treated suspension-cultured cells of French bean. An activation was observed within 10 min and was maintained for up to 4 h. The time course of activation was different from MAP kinase and casein kinase assayed in the same extracts. However, at 5 min after addition of elicitor there is a transient inactivation of the CDPKs before activation. This inactivation can be mimicked by adding forskolin to the cells 30 min before elicitation, which brings about changes in the cellular pH. Forskolin potentiates the oxidative burst when elicitor is subsequently added while the CDPK cannot be activated by elicitor upon forskolin treatment. In contrast, intracellular acidification brought about by forskolin brings about slight activation of MAPkinase. PMID- 12090627 TI - Probing the diversity of the Arabidopsis glutathione S-transferase gene family. AB - Glutathione S-transferases (GSTs) appear to be ubiquitous in plants and have defined roles in herbicide detoxification. In contrast, little is known about their roles in normal plant physiology and during responses to biotic and abiotic stress. Forty-seven members of the GST super-family were identified in the Arabidopsis genome, grouped into four classes, with amino acid sequence identity between classes being below 25%. The two small zeta (GSTZ) and theta (GSTT) classes have related GSTs in animals while the large phi (GSTF) and tau (GSTU) classes are plant specific. As a first step to functionally characterize this diverse super-family, 10 cDNAs representing all GST classes were cloned by RT-PCR and used to study AtGST expression in response to treatment with phytohormones, herbicides, oxidative stress and inoculation with virulent and avirulent strains of the downy mildew pathogen Peronospora parasitica. The abundance of transcripts encoding AtGSTF9, AtGSTF10, AtGSTU5, AtGSTU13 and AtGSTT1 were unaffected by any of the treatments. In contrast, AtGSTF6 was upregulated by all treatments while AtGSTF2, AtGSTF8, AtGSTU19 and AtGSTZ1 each showed a selective spectrum of inducibility to the different stresses indicating that regulation of gene expression in this super-family is controlled by multiple mechanisms. The respective cDNAs were over expressed in E. coli. All GSTs except AtGSTF10 formed soluble proteins which catalysed a specific range of glutathione conjugation or glutathione peroxidase activities. Our results give further insights into the complex regulation and enzymic functions of this plant gene super-family. PMID- 12090632 TI - Crossed and uncrossed stereoacuity at distance and the effect from heterophoria. AB - BACKGROUND: Previous studies have found that crossed disparities give better stereoacuity than uncrossed. When near phoria is considered, exophores had a better crossed stereoacuity and esophores had a better uncrossed stereoacuity. The current study investigated the effect of heterophoria on distant crossed and uncrossed stereoacuity. METHODS: Seventy-two subjects were recruited and their distant heterophoria measured. Heterophoria within two prism dioptres was considered as orthophoric. Esophoric group had esophoria greater than two prism dioptres. Exophoric group had exophoria greater than two prism dioptres. There were 40 orthophores, 23 exophores and nine esophores. Their stereoacuity was measured with a three-rod apparatus at 6 m. RESULTS: The mean crossed stereoacuity was 4.8" and uncrossed stereoacuity was 7.2" (t = -3.03, p < 0.01). The mean stereoacuity for orthophoric subjects was 5.31", 6.02" for exophores and 8.91" for esophores. The distant crossed stereoacuity is better than uncrossed stereoacuity in all three groups but the difference is only significant for the exophores. Exophoric subjects demonstrated a significant difference between crossed (4.10") and uncrossed (7.95") stereoacuity. CONCLUSIONS: Orthophores have the best stereoacuity, followed by exophores and esophores. Exophoric subjects have a better crossed than uncrossed stereoacuity. More esophoric subjects should be recruited to confirm the difference between crossed and uncrossed stereoacuity. PMID- 12090634 TI - Model of the optical system of the human eye during accommodation. AB - A model of the human eye with a gradient index crystalline lens is presented. The crystalline lens shape at different accommodation levels is described by a single function which is a combination of hyperbolic cosine functions and hyperbolic tangent functions. Using the experimental data published in the literature a model of the variations of the external lens shape was created. Formulae for the lens shape parameters and gradient index distribution for different accommodation levels are given. PMID- 12090633 TI - An assessment of the orthogonal astigmatism test for the subjective measurement of astigmatism. AB - The orthogonal astigmatism test (OAT) is a newly developed method for assessing ocular astigmatism. This study compares the OAT with the Jackson crossed-cylinder (JCC), Raubitschek arrow (RA) and the fan and block (FB) (fan chart). Fifty emmetropes or corrected spherical ametropes (<0.25 DC of astigmatism) were recruited, with a visual acuity of at least 6/5 in both eyes. Pseudo-astigmatism was induced by placing a +0.75 DC lens in front of the right eye at a random axis. With the left eye occluded, each subjective test was performed in a random order and the correcting lens power and axis noted. No significant difference was found in the ability of the OAT to locate the power and axis of the induced astigmatism over the JCC, RA and FB. In conclusion, the OAT offers the clinician an alternative method for locating and correcting ocular astigmatism. It has a similar but not superior accuracy to the other three subjective tests assessed. PMID- 12090635 TI - Diffraction haloes resulting from corneal oedema and epithelial cell size. AB - Corneal oedema was induced by exposing the corneas of nine subjects, covering an age range from 20 to 64 years, to distilled water for approximately 25 min. Measurement of the diameters of the associated diffraction haloes observed around a small 633 nm source allowed the size of the diffracting elements within each cornea to be estimated as about 10 microm, in good agreement with direct measurements by in vivo confocal microscopy of the subject's basal epithelial cells. PMID- 12090636 TI - MEM and Nott dynamic retinoscopy in patients with disorders of vergence and accommodation. AB - The accommodative response in 34 patients with accommodative and binocular disorders was assessed with two different techniques of dynamic retinoscopy used in clinical practice: monocular estimate method (MEM) and Nott retinoscopy. The data obtained by both techniques were compared, evaluating the correlation and agreement between them. Results showed that there were statistically significant differences between the techniques with MEM values being more plus than Nott ones. There was a high correlation between the two techniques (0.90) and the regression analysis indicated that a linear relationship existed between MEM and Nott dynamic retinoscopy, so that accommodative lag value for Nott dynamic retinoscopy would be calculated by dividing the MEM result by 2. However, although both retinoscopies were related, there was a lack of agreement between them (+/-0.53 D) indicating that the two methods are not interchangeable for clinical purposes. PMID- 12090637 TI - Image quality in wearers of a centre distance concentric design bifocal contact lens. AB - The optical performance of eyes wearing bifocal concentric contact lens was studied using the double-pass technique. Retinal image quality was measured for four subjects wearing CIBA Bisoft contact lenses presenting the central zone for correcting distance vision. Lenses with two different central optic zone diameter (COZD), 3.2 and 3.8 mm, were studied and the influence of pupil diameter and viewing distance were analysed. Results show that the best optical performance is obtained for far vision conditions when no lens is worn even if the pupil coverage by the COZD is complete. For near vision conditions, the optical performance when the lens is worn is, in general, better than when no lens is worn. When the lens is worn the best optical performance corresponds to a pupil diameter of 3 mm and far vision conditions. For this pupil diameter, variations in the situations analysed can be explained by changes in the percentage of pupil coverage corresponding to the far or the near vision zone of the lens. For a pupil diameter of 5 mm, the retinal image quality is more similar in all situations studied and pupil coverage alone cannot explain the results obtained and the influence of other parameters related to the design or contact lens fitting characteristics must be considered. PMID- 12090639 TI - The multifocal electroretinogram in central serous chorioretinopathy. AB - PURPOSE: To compare multifocal electroretinograms (ERGs) in control subjects and patients with central serous chorioretinopathy (CSC). METHOD: Eighteen eyes of control subjects and 15 eyes affected by CSC were tested with a VERIS Sciences 4.0 system. The average response densities and latencies of six annular retinal regions in control subjects were compared with those with CSC. RESULTS: The N1, P1 average response densities of the first and second annuli were decreased and the N1, P1 latencies of first to third rings were delayed in CSC. CONCLUSION: The multifocal ERG can be used to evaluate retinal dysfunction in CSC. PMID- 12090638 TI - Optimal green (red-free) digital imaging of conjunctival vasculature. AB - AIMS/BACKGROUND: Green illumination is commonly used to image vessels of the retina and conjunctiva. The purpose was to derive the best optical set-up for imaging vessels of the conjunctiva. METHODS: The concept of exposure density was used to predict a digital camera response to imaging vessels on a scleral background. Practical verification was performed to verify vessel contrast because of the difficulties in measuring the spectral components of the imaging system, such as the spectral reflectivity of vessels and sclera. Images of the same conjunctiva were repetitively taken through different coloured filters, using the Nikon FS-2 photo slit-lamp and recorded on different coloured channels of the Kodak DCS 100 digital camera. Gaussian blurred tubular models were fitted to densitometric profiles across three vessels from each image, allowing vessel contrast and width to be objectively measured. These measures were compared using different optical set-ups. RESULTS: Optimal exposure density calculations and vessel contrast was obtained with the xenon light source filtered with Wratten 99 (green) and Wratten 96 (neutral density, 0.2 log units) gelatine absorption filters using the green channel of the digital camera. This image set-up was associated with a 46% (99% CI 43-51%) to 64% (99% CI 58-72%) increase in contrast compared with vessels imaged without filtration, using the combined colour channel of the digital camera. Although differences in vessel widths resulted, absolute differences were marginal. CONCLUSION: With the increased use of digital imaging, and the need for image processing of vascular networks, image optimisation is beneficial. This study verified the optimal set-up for non invasively imaging vessels of the bulbar conjunctiva. PMID- 12090640 TI - The application of analysis of variance (ANOVA) to different experimental designs in optometry. AB - Analysis of variance (ANOVA) is the most efficient method available for the analysis of experimental data. Analysis of variance is a method of considerable complexity and subtlety, with many different variations, each of which applies in a particular experimental context. Hence, it is possible to apply the wrong type of ANOVA to data and, therefore, to draw an erroneous conclusion from an experiment. This article reviews the types of ANOVA most likely to arise in clinical experiments in optometry including the one-way ANOVA ('fixed' and 'random effect' models), two-way ANOVA in randomised blocks, three-way ANOVA, and factorial experimental designs (including the varieties known as 'split-plot' and 'repeated measures'). For each ANOVA, the appropriate experimental design is described, a statistical model is formulated, and the advantages and limitations of each type of design discussed. In addition, the problems of non-conformity to the statistical model and determination of the number of replications are considered. PMID- 12090641 TI - The stenopaeic slit: an analytical expression to quantify its optical effect in front of an astigmatic eye. PMID- 12090642 TI - Allergic rhinitis remains an important disease. PMID- 12090643 TI - Scope and impact of allergic rhinitis. AB - Allergic rhinitis is estimated to affect as many as 40 million people in the United States on a regular basis, and even more individuals who have occasional symptoms. The disease is associated with a considerable burden on the healthcare system, accounting for a total of $7.9 billion in direct and indirect costs in 1997, and with significant adverse effects on patients' quality of life, including disturbed sleep and impaired function at work and school. The pathophysiology of allergic rhinitis is complex, involving inflammatory mediators and immune cells that produce allergy symptoms via multiple mechanisms. The first principle of clinical management of patients with allergic rhinitis is avoidance of exposure to allergens, but this measure can be very difficult, and most patients require pharmacotherapy. Allergy vaccine therapy may be an appropriate and necessary option in selected patients with allergies refractory to other treatment modalities. PMID- 12090644 TI - Improved strategies and new treatment options for allergic rhinitis. AB - Primary principles relevant to the clinical management of allergic rhinitis include (1) avoidance of allergens and triggering factors, (2) use of appropriate pharmacotherapy, (3) evaluation regarding need for and appropriate use of immunotherapy, and (4) patient education and follow-up. Currently available pharmacotherapeutic options include oral and topical (intranasal) decongestants and corticosteroids, mast cell stabilizers, intranasal anticholinergics, and antihistamines. Future therapeutic options include leukotriene modifiers and anti IgE antibodies. PMID- 12090645 TI - DO questions need for proposed new tenets of osteopathic medicine. PMID- 12090648 TI - Biomechanics of growth and development in the healthy human infant: a pilot study. AB - Detailed measurements were acquired from 168 healthy subjects who were brought to the West Virginia School of Osteopathic Medicine's Robert C. Byrd Clinic in Lewisburg, WVa, during 1998 and 1999 by their parents for routine well-baby visits. Measurements of body-segment length, diameter, circumference, and skinfold thickness were taken at several segment locations. The center of volume was calculated for each body segment and was used as an approximation of center of mass. Findings from this study are strikingly similar to those found in the preceding long-term study by Turnquist and Wells of body-segment morphology, locomotion, and posture in the rhesus monkey (Macaca mulata). In human growth and development, the center of mass of each limb body segment migrates proximally, and more so in the more proximal segments. This trend is more pronounced in the lower extremities. Limb length and circumference increase with age in a linear pattern. These ontogenic changes are the consequence of the development of a propulsive hindlimb. This report is part of an ongoing study that will conclude in 2005 and that is intended to provide clinicians with detailed ontogenic information about changes in limb proportions in normal healthy children. PMID- 12090649 TI - Osteopathic manipulative treatment in conjunction with medication relieves pain associated with fibromyalgia syndrome: results of a randomized clinical pilot project. AB - Osteopathic physicians caring for patients with fibromyalgia syndrome (FM) often use osteopathic manipulative treatment (OMT) in conjunction with other forms of standard medical care. Despite a growing body of evidence on the efficacy of manual therapy for the treatment of selected acute musculoskeletal conditions, the role of OMT in treating patients with chronic conditions such as FM remains largely unknown. Twenty-four female patients meeting American College of Rheumatology criteria for FM were randomly assigned to one of four treatment groups: (1) manipulation group, (2) manipulation and teaching group, (3) moist heat group, and (4) control group, which received no additional treatment other than current medication. Participants' pain perceptions were assessed by use of pain thresholds measured at each of 10 bilateral tender points using a 9-kg dolorimeter, the Chronic Pain Experience Inventory, and the Present Pain Intensity Rating Scale. Patients' affective response to treatment was assessed using the Self-Evaluation Questionnaire. Activities of daily living were assessed using the Stanford Arthritis Center Disability and Discomfort Scales: Health Assessment Questionnaire. Depression was assessed using the Center for Epidemiological Studies Depression Scale. Significant findings between the four treatment groups on measures of pain threshold, perceived pain, attitude toward treatment, activities of daily living, and perceived functional ability were found. All of these findings favored use of OMT. This study found OMT combined with standard medical care was more efficacious in treating FM than standard care alone. These findings need to be replicated to determine if cost savings are incurred when treatments for FM incorporate nonpharmacologic approaches such as OMT. PMID- 12090651 TI - Prescription ergonomics: adding prevention to the diagnosis and treatment of work related musculoskeletal disorders. AB - The physician treating patients who have work-related musculoskeletal disorders (WMSDs) must educate the patient and employer about the nature of the problem and what to do about it. To do this effectively, diagnostic findings must be linked to prevention efforts in a way that is understandable for the physician, the patient, and the company's safety professional. Three skills are required. First, the physician must be able to recognize how the injury presentation reflects acute or chronic biomechanical stress patterns. Second, the physician must be able to recognize the work relationships and other elements that influence biomechanical stress. Third, the physician must have access to a mnemonic that enables him or her to analyze the work method and explain significant findings to the patient and employer. PMID- 12090650 TI - Osteoporosis prevention project: a model multidisciplinary educational intervention. AB - Osteoporosis is a systemic metabolic disease resulting in low bone mass, which increases the risk for fracture. Evidence suggests that lifestyle changes to prevent or delay development of osteoporosis should be implemented throughout the life span. The purpose of this study was to evaluate the effectiveness of a multidisciplinary primary osteoporosis prevention program for community-dwelling women aged 25 to 75 years to determine if osteoporosis prevention program participants (treatment group) increased their knowledge of osteoporosis, calcium intake, and exercise compared with a control group. Other outcomes included participants' willingness to adopt lifestyle changes and ability to view themselves as able to make behavioral changes. Subjects in the treatment group versus control subjects increased their knowledge of osteoporosis over time. At posttest, subjects in the treatment group were more likely to be planning to change calcium intake, and at follow-up, they were more likely to be changing their calcium intake. No other group differences were found between the two groups. These findings suggest that a multidisciplinary education program may have an impact on knowledge and behaviors that may help to delay the development of osteoporosis. PMID- 12090652 TI - Recommendations for the treatment of nicotine dependency. AB - The US Public Health Service (PHS) Clinical Practice Guideline Treating Tobacco Use and Dependence, published in June 2000, is a clinician-targeted resource that encourages physicians to apply evidence-based interventions when treating tobacco dependent patients. Issued by the PHS Surgeon General, the guideline recommends a new standard of care in treating tobacco dependency, the chief cause of preventable death and disease in the United States. It calls on clinicians to more consistently identify and evaluate tobacco-dependent patients and to provide treatment, motivation to quit, and referrals to smoking cessation specialists. The guideline also serves as a mandate for healthcare insurers to offer pharmacologic and behavioral treatment modalities as benefits to insured persons and to provide education and reimbursement for clinicians. PMID- 12090653 TI - Micro total analysis systems. 1. Introduction, theory, and technology. PMID- 12090654 TI - Micro total analysis systems. 2. Analytical standard operations and applications. PMID- 12090655 TI - Planar chromatography. PMID- 12090656 TI - Fiber-optic chemical sensors and biosensors. PMID- 12090657 TI - Current application of mass spectrometry to combinatorial chemistry. PMID- 12090658 TI - Atomic spectroscopy. PMID- 12090659 TI - Solid-state nuclear magnetic resonance. PMID- 12090660 TI - Environmental mass spectrometry: emerging contaminants and current issues. PMID- 12090661 TI - Mass spectrometry of chemical polymers. PMID- 12090662 TI - Thermal analysis. PMID- 12090663 TI - Chemometrics. PMID- 12090664 TI - Gas chromatography. PMID- 12090665 TI - Electrochemical sensors. PMID- 12090666 TI - Supercritical fluid and unified chromatography. PMID- 12090667 TI - Column liquid chromatography: equipment and instrumentation. PMID- 12090668 TI - Capillary electrophoresis for the analysis of biopolymers. PMID- 12090669 TI - Scanning probe microscopy. PMID- 12090670 TI - Chiral separations. PMID- 12090671 TI - Inductively coupled plasma mass spectrometry. PMID- 12090672 TI - X-ray spectrometry. PMID- 12090673 TI - Patient personality and mortality: a 4-year prospective examination of chronic renal insufficiency. AB - The present study examined the role of personality as a predictor of mortality among patients with chronic renal insufficiency. A prospective evaluation of the influence of personality on patient survival was conducted over an average 49 month period. Cox regression was used to evaluate the effects of 5 dimensions of personality in a sample of 174 patients (100 male and 74 female). At follow-up, 49 patients had died. Significant demographic and clinical predictors of survival included age, diabetic status, and hemoglobin level. After these predictors were controlled for, 2 personality traits, conscientiousness and neuroticism, predicted patient mortality. Patients with high neuroticism scores had a 37.5% higher estimated mortality rate. Patients with low conscientiousness scores had a 36.4% increased mortality rate. PMID- 12090674 TI - Stress-related immune changes in middle-aged and older women: 1-year consistency of individual differences. AB - This study reviews prior research and reports longer-term consistency of stress related immune variables in middle-aged and older women who performed mental math and speech tasks 2 times 1 year apart. Leukocyte subsets, mitogen-induced lymphocyte proliferation, and natural killer cell activity were measured at baseline, after tasks, and after 30-min recovery. Epstein-Barr virus (EBV) antibody titers were assessed at baseline. Pearson coefficients and standardized maximum-likelihood estimates of year-to-year covariances for leukocyte subsets and EBV titers showed moderately high to high baseline and posttask consistency and lower recovery consistency; consistency for other functional immune assays and reactivity scores for all variables was moderate to low. Results support longitudinal study of psychosocial context effects on tonic immune function and posttask scores. PMID- 12090675 TI - Acute nicotine reinforcement, but not chronic tolerance, predicts withdrawal and relapse after quitting smoking. AB - Little research has examined the association of tobacco dependence with nicotine tolerance or reinforcement in a clinical sample. Smokers preparing to quit smoking participated in laboratory sessions to assess nicotine tolerance on subjective, cardiovascular, and performance measures and to assess nicotine reinforcement using a choice procedure. Participants were then provided with individual counseling (but no medication), made a quit attempt, and were followed for 1 year to determine clinical outcome, as determined by postquit withdrawal and days to relapse. Nicotine tolerance was unrelated to either withdrawal or relapse. However, acute nicotine reinforcement was significantly related to both greater withdrawal and faster relapse. Results challenge the common assumption that nicotine tolerance is closely related to dependence but suggest that nicotine reinforcement may have theoretical and clinical significance for dependence. PMID- 12090676 TI - Health stresses and depressive symptomatology in the elderly: the importance of health engagement control strategies. AB - The study presents cross-sectional (N = 127) and longitudinal (n = 111) analyses examining relations between health engagement control strategies (HECSs), depressive symptoms, and health stresses in elderly individuals. HECS was measured as people's behavioral and cognitive investments toward attaining health goals. HECS was related to low levels of depressive symptoms, particularly among people experiencing acute physical symptoms. Moreover, HECS predicted reduction of depressive symptoms over time, and depressive symptomatology predicted negative change in HECS. The findings show that active investments of HECSs significantly moderate the negative affective consequences of health threats. Individuals who are characterized by low levels of HECS and high levels of depressive symptoms may be at increased risk of accelerated decline in their physical and mental health. PMID- 12090677 TI - The influence of peers on young adult substance use. AB - Data collected from 294 young adults, ages 19 to 25, and both a same- and an opposite-gender best friend or mate across 3 annual assessments were analyzed to examine the similarity to and influence of the peer on the young adult's substance use. The authors found similarity across time between both peers and the young adult in cigarette use, alcohol use, binge drinking, and, in most cases, marijuana use. In prospective analyses, peer use predicted young adult cigarette use, binge drinking, and problem use by the young adults. Results were generally consistent across gender and for both same- and opposite-gender peers. Findings emphasize peer influence contribution to young adult substance use and suggest the design of interventions that involve both young adults and their peers. PMID- 12090678 TI - Menstrual cycle, beta-endorphins, and pain sensitivity in premenstrual dysphoric disorder. AB - This study examined pain sensitivity and pain modularity mechanisms (e.g., beta endorphin levels, blood pressure) in women with premenstrual dysphoric disorder (PMDD; n = 27) and healthy controls (n = 27) during the follicular and luteal phases of the menstrual cycle. Physiological measures were taken during rest and ischemic pain testing. In both cycle phases, PMDD women (a) displayed lower resting cortisol and beta-endorphin levels and (b) exhibited shorter pain threshold and tolerance times and greater pain unpleasantness ratings during pain. PMDD women also reported greater pain unpleasantness and intensity and had lower beta-endorphin levels in their luteal phase and tended to display higher blood pressure levels at rest and during pain testing. Results suggest that endogenous opioids may be pathophysiologically relevant to PMDD and that the hypothalamic-pituitary-gonadal axis may modulate pain sensitivity in PMDD. PMID- 12090679 TI - Effects of gender in social control of smoking cessation. AB - This study of 93 men and 117 women smokers during an ongoing quit attempt examined the roles of gender and social network influences on quitting. For men, social influences appeared to positively affect their ability to reduce their smoking but were less effective for women. Specifically, increased reports of a spouse or partner's influence, and family and friends' influence, were associated with greater reductions in men's smoking 2 days and 4 months post quit date, respectively. In contrast, for women, greater reports of spouse or partner influence and of family and friends' influence were associated with smaller reductions in smoking. Sex differences in social control strategies and perceived autonomy supportiveness of those strategies are discussed as possible explanations for these results. PMID- 12090680 TI - Older adults' recovery from surgery for osteoarthritis of the knee: psychosocial resources and constraints as predictors of outcomes. AB - The authors investigated psychosocial resources (positive support, active coping) and psychosocial constraints (negative support, avoidant coping) as predictors of improvement in the health of 63 older adults undergoing surgery for osteoarthritis of the knee. Following surgery, patients and social partners are motivated both to strive for the appetitive goal of recovery (approach), and to protect themselves from pain and impairment (avoidance). The authors assessed resources and constraints 6 weeks after surgery as predictors of outcomes (improvement in knee pain, knee functioning, and psychological well-being) 6 months after surgery. The constraints patients encountered early in recovery were strong predictors of poor recovery. Although resources were associated with some improvement, these effects were largely accounted for by constraints. PMID- 12090681 TI - Psychosocial antecedents of tobacco chipping. AB - The authors examined adolescent antecedents and adult correlates of tobacco "chipping" compared with heavy smoking, experimental smoking, and nonsmoking in a representative community sample. As adolescents, future "chippers" had some smoking risk factors (attitudes, health beliefs, smoking intentions, and tolerance for deviance) and several protective factors (high values for academic success, internal locus of control, supportive relationships, and little smoking among peers and parents). As adults, risk factors included lessened belief in nicotine's addictiveness, lower conscientiousness, higher extraversion, and lower positive affect and life satisfaction; protective factors included high levels of educational attainment and employment and low levels of negative affect and stress (men only). Thus, chippers experience a complex mixture of risk and protective factors for smoking. PMID- 12090682 TI - Effect of a behavioral self-regulation intervention on patient adherence in hemodialysis. AB - The present study examined the efficacy of a behavioral intervention designed to increase adherence to fluid-intake restrictions among hemodialysis patients. Twenty intervention-group patients were compared with 20 matched control patients on an indicator of fluid-intake adherence at 3 time points. The Group x Time interaction was significant, indicating that patients in the 2 groups exhibited a differential pattern of change in fluid-intake adherence across the follow-up period. The intervention and control groups did not differ significantly in terms of adherence at the initial postintervention period but did differ at the 8-week follow-up. The observed group differences were, in part, due to a trend toward increasingly better adherence in the intervention group and poorer adherence in the control group across the follow-up period. PMID- 12090683 TI - Internet use and social support in women with breast cancer. AB - Many Web sites offer information to breast cancer patients, who are increasingly using these sites. The authors investigated the potential psychological benefits of Internet use for medical information by breast cancer patients. Of the 251 women approached, 188 were successfully interviewed (74.9%). Forty-two percent used the Internet for medical information related to breast health issues and did so for an average of 0.80 hr per week. The Interpersonal Support Evaluation List and the UCLA Loneliness Scale, with results controlled for covariates, showed that Internet use for breast health issues was associated with greater social support and less loneliness than Internet use for other purposes or nonuse. Breast cancer patients may obtain these psychological benefits with only a minimal weekly time commitment. PMID- 12090684 TI - Caregiver models of self and others, coping, and depression: predictors of depression in children with chronic pain. AB - In a sample of 59 chronically ill pediatric patients and their maternal caregivers, both child-reported pain and caregiver-reported depression predicted child-reported depression. Results further suggested that the association between pain and depression in children is ameliorated by caregiver coping strategies and that how caregivers cope is a function of their attachment-related representations of the self and others. Caregivers with a negative model of the self were more depressed. and those with a negative model of others were more prone to use avoidant coping strategies, and, in turn, to be more depressed. However, the extent to which caregivers with negative models of self used more avoidant and less approach coping appeared to depend on whether they perceived that others were likely to respond to their needs. PMID- 12090685 TI - Crossing the quality chasm: building a safe nutrition system. AB - Recent reports to Congress and the public from the Institute of Medicine underscore concern about the quality of healthcare in America. The nutrition community has focused most of its attention on the report titled The Role of Nutrition in Maintaining Health in the Nation's Elderly, which evaluated nutrition services coverage for the Medicare population. Of equal importance was the recent publication of two reports from the Committee on Quality of Health Care in America: To Err is Human--Building a Safer Health System and Crossing the Quality Chasm--A New Health System for the 21st Century. IV nutrition support was a breakthrough in medical care that has become a standard tool in treating patients who cannot eat for prolonged periods of time. It is also a medical treatment that can result in harm to patients. As problems with patient safety associated with the use of IV nutrition were documented, safer methods to deliver this life-saving form of treatment were developed and evaluated. Although an interdisciplinary team approach has been shown to be the safest way to administer IV nutrition, this system is costly and not universally used. Alternatives to the interdisciplinary team approach should be evaluated to assure that patients receive optimum nutrition care. The tools that can be used to improve patient safety include self-assessment of practitioners who routinely use nutrition support in their practice, curricular-based continuing education programs, board certification in nutrition support practice, and the use of clinical guidelines to assist in making clinical decisions. By developing and promoting these tools, A.S.P.E.N. is committed to building a safe nutrition system so every patient receives optimal nutrition care. PMID- 12090687 TI - Early enteral nutrition-the unanswered Ws. PMID- 12090686 TI - Specific intraluminal nutrients alter mucosal blood flow during gut ischemia/reperfusion. AB - BACKGROUND: We have previously demonstrated in a rodent model the differential effects that specific intraluminal nutrients exert on gut ischemia/reperfusion (I/R) injury. Alanine was shown to amplify, whereas glucose protected against, gut I/R injury and associated gut dysfunction. The objective of this study was to determine whether these specific nutrients are associated with alterations in mucosal perfusion during gut I/R. METHODS: Sprague-Dawley rats had either a laser doppler probe or a tonometer inserted into a jejunal sac filled with either 10 mmol/L alanine, glucose, or mannitol (osmotic control) followed by 60 minutes of superior mesenteric artery occlusion and 60 minutes of reperfusion. Laser doppler mucosal blood flow and regional PCO2 (PrCO2) measurements were obtained. RESULTS: Mucosal blood flow was significantly increased during both ischemia and reperfusion when intraluminal glucose was present compared with intraluminal alanine. Blood flow changes were reflected by lower jejunal PrCO2 measurements with intraluminal glucose compared with intraluminal alanine. CONCLUSIONS: Intraluminal glucose can augment mucosal blood flow during gut I/R and may explain the protective effect we previously observed. PMID- 12090688 TI - Evaluation of stress factors and body weight adjustments currently used to estimate energy expenditure in hospitalized patients. AB - BACKGROUND: The purpose of this study was to critically examine current approaches for estimating energy expenditure and requirements of hospitalized patients requiring nutrition support. METHODS: All indirect calorimetry tests performed by our adult nutrition support service from 1991 to 2000 were reviewed. Stress factors were calculated as the measured energy expenditure divided by the predicted energy expenditure using the Harris-Benedict equation. Various methods for adjusting the body weights of obese subjects for use in the Harris-Benedict equation were evaluated. RESULTS: The average stress factor for these hospitalized patients was 1.25, and there were no differences in the stress factors between men and women. For obese subjects, using an adjusted body weight equal to ideal body weight plus 50% of the difference between ideal and actual body weight in the Harris-Benedict equation gave stress factors comparable with normal weight subjects. For underweight subjects, using the actual rather than ideal body weight gave stress factors that were most similar to normal weight patients. Disease-specific stress factors were calculated and compared with literature values. Mechanical ventilation, recent feeding, fever, and restlessness during the indirect calorimetry measurement increased the measured energy expenditure. CONCLUSIONS: Our findings provide nutrition support services with updated information on disease-specific stress factors that can be used for estimating energy expenditure in hospitalized patients. An adjusted body weight equal to ideal body weight plus 50% of the excess body weight should be used for estimating the energy requirements of obese patients requiring nutrition support. PMID- 12090690 TI - Enteral versus parenteral nutrition: the patient's preference. AB - BACKGROUND: Given that, in most clinical circumstances, enteral nutrition (nasal gastric tube feeding) may not necessarily be superior to parenteral nutrition (IV feeding), consideration of patient preference should be included in decisions on the method of feeding. OBJECTIVES: To evaluate patient preference for nasal gastric versus IV feeding. METHODS: We administered a written questionnaire to 101 hospitalized oncology patients and 98 outpatients without gastrointestinal illness (controls). Statistical analyses included comparisons of group means with Student t tests, comparisons of proportions with chi2 tests, and logistic regression analysis. RESULTS: Demographic variables of the 2 groups were comparable. In both groups, most individuals preferred IV to tube feeding. Preference for IV or tube feeding was related to patient perception of the comfort of these interventions. In logistic regression analyses, the strongest influences on preference were age and perceived comfort of IV feeding. Older individuals preferred IV to tube feeding. Gender, education level, physician's recommendation, and cost did not influence patient preference. CONCLUSIONS: Most patients prefer IV to nasal gastric feeding. Awareness of this patient preference is helpful in making decisions regarding the method of nutrient delivery. PMID- 12090689 TI - Evaluation of board certification in nutrition support pharmacy. AB - BACKGROUND: Credentialing of healthcare professionals is a topic that has received increasing attention. The Board of Pharmaceutical Specialties (BPS) has recognized nutrition support pharmacy as a specialty for more than 10 years. There has recently been concern about the decline in the number of pharmacists seeking board certification or recertification as specialists in nutrition support and changes in the job responsibilities of nutrition support pharmacists. These factors have resulted in a need to evaluate the current system of certification of nutrition support pharmacists. METHODS: A national survey was developed and sent to pharmacist members of the American Society for Parenteral and Enteral Nutrition (A.S.P.E.N.) and members of the Home Care Section of the American Society of Health-System Pharmacists (ASHP). The objectives of this study were to describe the activities of nutrition support pharmacists, determine the benefits and barriers to board certification, and assess the current system of certification. RESULTS: Two hundred and fifty-eight of 486 surveys were returned for an overall response rate of 53%. There has been a decrease in the amount of professional time devoted to nutrition support activities. The most highly ranked benefit to certification was peer recognition. The most highly ranked barrier to certification was the examination cost. CONCLUSIONS: There is satisfaction with the current system; however, alternative methods were supported if the current method cannot sustain itself There is a need to evaluate the cost of the examination and increase marketing efforts to candidates for whom the examination is intended. PMID- 12090691 TI - Understanding and augmenting human intestinal adaptation: a call for more clinical research. AB - Since its description in the early 20th century, the phenomenon of intestinal adaptation has been the focus of considerable interest. However, there is a paucity of evidence documenting the significance of this phenomenon in man. Indeed, our understanding of intestinal adaptation is based almost exclusively on extrapolations from studies of laboratory rodents. Given the possibility of pharmacologically modifying select components of adaptation as a therapeutic strategy for patients with intestinal insufficiency, a clear understanding of human intestinal adaptation is of paramount importance. PMID- 12090692 TI - Is gold always present in a "gold standard"? PMID- 12090693 TI - Vitamins and trace elements in TPN. PMID- 12090694 TI - Unusual aspects of GH re-testing. PMID- 12090695 TI - Bone status in children and adolescents with growth hormone deficiency: effect of growth hormone treatment. PMID- 12090697 TI - Is the impact of GH deficiency on the heart dependent on time of onset? PMID- 12090696 TI - The consequences of discontinuing GH after linear growth is completed. PMID- 12090698 TI - Is final height the only target of paediatric human growth hormone treatment? PMID- 12090700 TI - Childhood-onset growth hormone deficiency: reassessment of GH status after completion of growth. PMID- 12090699 TI - Effects of GH replacement in young patients with childhood onset GH deficiency. PMID- 12090701 TI - New-generation pulsed carbon dioxide laser: comparative effects on vocal fold wound healing. AB - We investigated wound healing of vocal fold mucosal excisions using a canine model, comparing cold steel, the continuous wave (CW) carbon dioxide (CO2) laser, and a new, microsecond-pulsed CO2 laser. The thermal injury to the surrounding normal tissues produced by the laser increases the risk for scar formation and poor functional voice outcome. This injury may be reduced with a pulsed CO2 laser. Acute, 2-week, and 6-week studies of 15 dogs were made with blinded observers and nonparametric statistical analyses. Histologic comparison showed less thermal injury and scar formation in the vocal folds treated with the pulsed CO2 laser than with the CW CO2 laser. Functional outcomes as studied with laryngeal videostroboscopy revealed better vibratory characteristics with the pulsed CO2 laser as compared with the CW CO2 laser. No differences were seen on these measures between the pulsed laser and cold techniques. Pulsed CO2 lasers may offer the ease of laser laryngology and the superior wound healing of cold steel. PMID- 12090702 TI - Cricothyroid approximation to elevate vocal pitch in male-to-female transsexuals: results of surgery. AB - Functional changes in voice production, including pitch elevation, may help male to-female transsexuals assume a new gender identity. To date, there has been a paucity of objective data on the effectiveness of pitch-raising methods. Acoustic data were gathered with regard to preoperative and postoperative changes in pitch, pitch range, and perturbation in 20 patients after cricothyroid approximation. Subjective data were gathered by means of a mailed questionnaire. The mean follow-up time was 22 months. The speaking fundamental frequency was raised by half an octave without any significant changes in perturbation. The lower and upper limits of pitch range both increased by an average of 4 semitones. There was some decline over time of the lower pitch range toward preoperative levels, but the upper pitch range remained elevated. The majority of the patients were satisfied with the results of surgery and felt their voices to be more feminine. PMID- 12090703 TI - 585-nm pulsed dye laser treatment of glottal papillomatosis. AB - Treatment of recurrent respiratory papillomatosis of the glottis is often challenging. The surgeon and patient must cooperatively balance decisions regarding airway safety, effects of multiple general anesthesias, employment disturbance, and vocal dysfunction. A pilot study was done in 41 adult cases (23 patients; 78 vocal folds) without complication to evaluate the effectiveness of a 585-nm pulsed dye laser (PDL; 450-micros pulse width; fluence of 38 to 255 J/cm2; 1- to 2-mm spot size) in the treatment of this disorder. Thirty-seven of the 41 cases (90%) were bilateral disease. Twenty-six of the 41 cases (63%; including 20 cases with involvement of the anterior commissure) were treated by bilateral photocoagulation of the lesions' microcirculation without microflap resection of tissue. Clinical observation revealed that irradiated but unresected disease involuted without development of an anterior commissure web. In the initial 13 of the 41 cases (32%), PDL treatment was followed by cold instrument microflap resection. The PDL enhanced the epithelial excision by improving hemostasis and by creating an optimal dissection plane between the basement membrane and the underlying superficial lamina propria. The PDL at 585 nm was less effective in the management of exophytic lesions because of its limited depth of penetration (approximately 2 mm). In this initial trial, the PDL was a relatively safe and efficacious treatment for glottal recurrent respiratory papillomatosis. Since the lesions involute without complete resection of the diseased epithelium, the anterior commissure can be treated to minimize the number of procedures. To study patterns of recurrence will require longer follow-up. PMID- 12090704 TI - Surgical voice rehabilitation after laser surgery for glottic carcinoma. AB - Transoral laser surgery today is the mainstay of treatment for T1 and T2 glottic carcinoma. The vocal ability remains sufficient in the majority of patients. However, in some cases, a significant glottic gap may persist, leading to poor voice quality. We report a special technique of medialization thyroplasty using autologous cartilage specifically adapted for vocal rehabilitation after laser resection. Six patients with a significant glottic gap following laser surgery were treated. For vocal rehabilitation, a special medialization technique was performed. The superior rim of the thyroid cartilage of the resected side was exposed. A 1 x 2-cm piece of cartilage was harvested and reimplanted into a subperichondrial pouch created on the inner side of the thyroid cartilage. When phonation was optimal, this cartilaginous strut was sutured and/or glued in place. In all 6 cases, the vocal function improved significantly. The dysphonia index (0 = normal, 3 = aphonia), which includes objective parameters as well as expert voice ratings and the patient's perception, increased by 1.1 on average (range, 0.4 to 1.6). The results have been lasting. The established medialization techniques are of limited value in a larynx scarred by laser surgery. Injection augmentation is often futile because the tight scar tissue does not lend itself to augmentation. Implantation of nonorganic material may cause problems if revision surgery for tumor recurrence should become necessary or if the implant protrudes into the scarred endolarynx. The technique reported avoids these pitfalls and leads to voice quality improvement comparable to that of established medialization procedures. PMID- 12090705 TI - Electrically stimulated glottal opening combined with adductor muscle botox blockade restores both ventilation and voice in a patient with bilateral laryngeal paralysis. AB - The purpose of this study was to determine whether paced electrical stimulation of the posterior cricoarytenoid muscle with an implantable device could restore ventilation in a patient with bilateral vocal fold paralysis without disturbing voice. In the first US case of a multi-institutional study, this patient was implanted with an Itrel II stimulator (Medtronic, Inc). In monthly postoperative sessions over an 18-month period, an effective stimulus paradigm was derived, the magnitude of stimulated vocal fold abduction and ventilation was measured, and perceptual judgments of voice quality were made. After identification of optimum parameters, posterior cricoarytenoid muscle stimulation produced a moderately large vocal fold abduction of 4 mm, but only marginal improvement in mouth ventilation, with no change in voice quality. After adductor muscle blockade with botulinum toxin, the patient's voice improved with increased phonatory airflow, but ventilation through the passive airway was still inadequate. However, by combining these two therapeutic strategies, dynamic abduction increased to 7 mm, ventilation through the mouth surpassed that through the tracheotomy (allowing decannulation), and voice quality was restored to normal. PMID- 12090706 TI - Immortalization of normal adult human middle ear epithelial cells using a retrovirus containing the E6/E7 genes of human papillomavirus type 16. AB - A human middle ear epithelial cell line (HMEEC-1) was established using human papillomavirus E6/E7 genes. HMEEC-1 has remained morphologically and phenotypically stable, even after 50 passages. The cells are anchorage-dependent and nontumorigenic when injected into nude mice. This cell line thus provides a new tool for the study of normal cell biology and the pathological processes associated with the epithelial cells of the middle ear in otitis media. HMEEC-1 will also be useful in the search for new drugs and biological agents for the treatment of otitis media. PMID- 12090707 TI - Electrically stimulated olfactory evoked potential in olfactory disturbance. AB - Olfactory evoked potential is considered a useful method of electrophysiological olfactometry for the diagnosis of olfactory disturbance. However, electrophysiological olfactometry is not as widely used as electrophysiological audiometry, such as the auditory brain stem response, because odor stimulation is difficult to perform. In contrast, electrical pulse stimulation is easy to perform, and its evoked potential is also easily recorded by the averaging method. We recorded olfactory evoked potentials from the scalp produced by electrical stimulation on the olfactory mucosa and investigated the relationship between this electrical olfactory evoked potential (EOEP) and the results of Toyoda and Takagi's perfumist's strip method (T&T) olfactometry, which is a standard Japanese means of psychophysical olfactometry. In EOEP-detectable cases, the detection and cognitive thresholds of T&T olfactometry were 1.32+/-1.99 (mean +/- SD; n = 10) and 3.02+/-1.64 (n = 10), respectively. But in the undetectable cases, the thresholds were 4.67+/-2.03 (n = 8; 5.8 means off the scale) and 5.80+/-0.00 (n = 8), respectively. The differences between the T&T thresholds of EOEP-detectable and -undetectable cases were significant. We concluded that EOEP is suitable for electrophysiological olfactometry. PMID- 12090708 TI - Results of fascial augmentation in unilateral vocal fold paralysis. AB - The objective of this study was to evaluate the effects on voice quality of augmentation by injection of minced fascia in patients with unilateral vocal fold paralysis. Preoperative and postoperative voice samples from 14 patients (6 men and 8 women; mean age, 59 years) were analyzed by computerized acoustic analysis and blinded perceptual evaluation. Statistically significant improvements were seen in perturbation measurements (jitter and shimmer), noise-to-harmonics ratio, and maximum phonation time. A panel of evaluators rated 10 of the 14 postoperative voices as normal or near-normal. Injection laryngoplasty with minced fascia offers a new, effective, well-tolerated, and inexpensive method to medialize a paralyzed vocal fold. The graft seems to survive well, as indicated by good vocal results with a follow-up ranging from 5 to 32 months. PMID- 12090709 TI - Functional anatomy of levator veli palatini muscle and tensor veli palatini muscle in association with eustachian tube cartilage. AB - The anatomic relationships among the levator veli palatini muscle (LVPM), the tensor veli palatini muscle (TVPM), and the eustachian tube (ET) cartilage were investigated by computer-aided 3-dimensional reconstruction and measurement methods. The study used 13 normal temporal bone-ET specimens obtained from 13 individuals (range of age at death, 3 months to 88 years). This study revealed several anatomic features of the anterior cartilaginous portion of the ET. First, the LVPM is always located inferolateral to the inferior margin of the medial lamina (ML) of the ET cartilage. Second, the LVPM has a large cross-sectional area throughout the extent of the anterior cartilaginous portion of the ET. Third, although the LVPM lies close to the ML of the ET cartilage (0.44+/-0.16 mm in children and 1.02+/-0.58 mm in adults), there is no region of attachment. Finally, the TVPM is not attached to the lateral lamina (LL) of the ET cartilage of the anterior quarter of the cartilaginous portion. Accordingly, it could be assumed that the most anterior cartilaginous portion of the ET is opened primarily by the contraction of the LVPM, which causes a superior-medial rotation of the ML. Furthermore, since the contraction time of the LVPM is reported to be longer than that of the TVPM, the anterior cartilaginous portions of the ET may remain open, even after the middle to posterior cartilaginous portions are closed after relaxation of the TVPM. This process would produce a pumping action of the ET in the direction from the middle ear to the pharyngeal side. The pumping function may be beneficial to clearance of the middle ear. PMID- 12090710 TI - Videostrobolaryngoscopy of mucus layer during vocal fold vibration in patients with laryngeal tension-fatigue syndrome. AB - The mucus layer on the vocal folds was examined by videostrobolaryngoscopy in patients with laryngeal tension-fatigue syndrome, a chronic functional dysphonia due to vocal abuse and misuse. Besides the findings in previous reports (such as abnormal glottal closure, phase or amplitude asymmetry, and the irregular mucosal wave), the vocal folds during vibration had an uneven mucus surface. The occurrence of an uneven mucus layer on vocal folds was significantly greater in subjects with this voice disorder (83% or 250 of 301 patients in this series) than in those without voice disorders (18.5% or 5 of 27). The increase of mucus viscosity, mucus aggregation, and the formation of rough surfaces on the vocal folds alter the mechanical properties that contribute to vibration of the cover of the vocal folds, and thereby worsen the symptoms of dysphonia in patients with laryngeal tension-fatigue syndrome. PMID- 12090711 TI - Electronic analysis of intrinsic laryngeal muscles in canine sound production. AB - This study explores the relationship between voice production and intrinsic laryngeal muscle (ILM) activities as expressed by orderly recruitment of their specific motor units. In 5 dogs, both the recurrent laryngeal nerve (RLN) and the vagus nerve (cranial nerve X) were stimulated via tripolar electrodes with stimulating frequencies (Fs) of 10 to 60 Hz and 0 to 7 mA during application of symmetric 600 Hz, 7 to 0 mA blocking currents. The fundamental frequency (Fo) and the intensity (I) of sounds generated by tracheal insufflation of humidified air were recorded while electromyograms of the cricothyroideus (CT), thyroarytenoideus (TA), and posterior cricoarytenoideus (PCA) were obtained via surface electrodes. Contractions of the CT were concurrently induced by stimulating the superior laryngeal nerve (SLN). The recruitment rates were highly specific and were affected by which nerve was stimulated. For the RLN, PCA ramping was lowest for Fs of < or =50 Hz. For Fs of 10 to 30 Hz, the recruitment rate of the TA was significantly steeper than that for the other ILMs, and the CT had the highest rate for Fs of 40 to 50 Hz. Conversely, for the vagus nerve, PCA recruitment was highest for Fs of > or =30 Hz. The average Fo was significantly higher with the RLN than with the vagus nerve. When the TA recruited faster than the CT (ie, via the RLN, but not the vagus nerve), the Fo was higher. While only CT ramping was significantly related to changes in sound intensity, there was a trend toward a decrease when PCA ramping was higher than CT ramping, as occurred when only the vagus nerve was stimulated. Stimulation of the SLN always increased Fo and loudness. We conclude that changes in Fo occur mainly through RLN-mediated CT and TA contraction. Loudness is controlled by the CT. The PCA exerts reciprocal coupling on both functions via the vagus nerve, and they are boosted across the board by the SLN. These findings may allow artificial manipulation of voice. PMID- 12090713 TI - Sensorineural hearing loss with intracranial venous malformations in Klippel Trenaunay syndrome. PMID- 12090712 TI - Therapeutic methods used for otolaryngological problems during the Byzantine period. AB - Evidence of herbal, animal, and chemical substances from the natural world used in medicines for otolaryngological problems, including opium, hyoscyamus, barley, honey, dried beans and peas, olives, fruits, Agaricus, castoreum, cassia, and afronitron, was traced in the Byzantine medical treatises, mainly from the 4th century AD to the 15th century AD. The texts of Antyllus, Orivasios of Pergamos, Aetios of Amida, Alexander of Tralles, Paul Aeginitis, Leon Iatrosophistis, Theophanis Nonnos, Nickolaos Myrepsos, Michael Psellos, and others strongly suggest the influence of ancient Greek and Roman medicine, but at the same time stress original medical thought. The main otolaryngological problems encountered in that period were loss of hearing, purulent otitis, rupture of the tympanic membrane, pharyngitis, laryngitis, rhinitis, acute tonsillitis, seasickness, vertigo, fracture of the nose, and cancers of the ear, larynx, nose, and oral cavity. The tradition stating that remedies were the final products of substance combinations, started in the classical period (5th and 4th centuries BC), is presented clearly and in detail in Byzantine prescriptions related to otolaryngology. PMID- 12090714 TI - Lack of evoked laryngeal electromyography response in patients with a clinical diagnosis of vocal cord paralysis. PMID- 12090715 TI - Involvement of level I neck lymph nodes in advanced squamous carcinoma of the larynx. PMID- 12090716 TI - Postoperative cognitive dysfunction versus complaints: a discrepancy in long-term findings. AB - This review describes the discrepancy in findings between postoperative cognitive performance and postoperative cognitive complaints long time after an operation under general anesthesia. Shortly (from 6 hr to 1 week) after an operation a decline in cognitive performance is reported in most studies. However, long time (from 3 weeks to 1-2 years) after an operation this is rarely found although some patients are still reporting cognitive complaints. In general this kind of research is suffering from severe methodological problems (use of insensitive tests, lack of control groups, lack of parallel tests, different definitions of cognitive decline). However, these problems cannot totally explain the discrepancy in findings in the long term. Thus, there are patients who have persistent cognitive complaints long time after an operation, that cannot be measured with cognitive tests. More psychological factors such as fixation on short-term cognitive dysfunction, mood, coping style, and personality are possible explanations for these cognitive complaints in the long term. As a consequence, these factors should be a topic in future research elucidating the persistence of these cognitive complaints long time after an operation under general anesthesia. PMID- 12090717 TI - Neurocognitive functioning in posttraumatic stress disorder. AB - This paper reviews the literature on performance on standard neuropsychological tests among individuals with posttraumatic stress disorder (PTSD). Of 19 studies, 16 reported impairment of attention or immediate memory (or both); however, most of these studies included PTSD patients with significant psychiatric comorbidity, so that the extent to which the observed deficits are specifically attributable to PTSD remains unclear. Other potential confounds, including medical illness, substance abuse, and motivational factors, further preclude definitive conclusions at present. Results of structural and functional neuroimaging studies of PTSD are also summarized. Two studies have reported correlations between hippocampal volume and cognitive findings in PTSD patients; functional studies have indicated specific findings in limbic regions, although the relationship of these results to neuropsychological performance remains to be explored. PMID- 12090719 TI - Antibiotics for the treatment of onchocerciasis and other filarial infections. AB - More effective drugs are needed for the treatment of human filarial diseases and the elimination of these infections as a public health problem. The drugs must either kill or sterilize adult worms. The relevant filariae, Onchocerca volvulus, Wuchereria bancofti and Brugia species, harbor rickettsial endoboacteria of the genus Wolbachia as symbionts. Animal experiments have shown that the elimination of these endobacteria causes inhibition of embryogenesis, and with Onchocerca ochengi a macrofilaricidal effect. Trials with human onchocerciasis patients using doxycydine demonstrated a long-term sterilizing activity, opening up a new strategy for the control of filarial infections. Indications of antiwolbachial therapy against onchocerciasis are discussed. PMID- 12090720 TI - Topical treatment for cutaneous leishmaniasis. AB - Parenteral pentavalent antimonials remain the standard therapy for cutaneous leishmaniasis. More effective and patient-compliant topical treatments are an important alternative treatment for the localized self-limiting forms of this disease. Two paromomycin ointments are commercially available but their use is limited by either toxicity or lack of efficacy. Other topical formulations have been in clinical trials, but many results have been equivocal and no major breakthroughs have been achieved. The focus of this review is on recent developments in the field of topical treatment for cutaneous leishmaniasis and rational approaches to enhance topical drug absorption. PMID- 12090721 TI - Artemotil Artecef. AB - The ethyl ether derivative of artemesinin, artemotil, has been developed by Artecef for the potential treatment of malaria. Adult and pediatric intramuscular formulations of the product were filed for approval in The Netherlands in January 1997 [236187]; registration for the two formulations followed in May 2000 [401672]. PMID- 12090718 TI - A review of diagnostic techniques in the differential diagnosis of epileptic and nonepileptic seizures. AB - The diagnosis of psychogenic nonepileptic seizures (PNES) is complex. Long-term electroencephalogram monitoring with video recording (video EEG) is the most common method of differential diagnosis of epilepsy and PNES. However, video EEG is complex, costly, and unavailable in some areas. Thus, alternative diagnostic techniques have been studied in the search for a diagnostic method that is as accurate as video EEG, but more cost effective, convenient, and readily available. This paper reviews the literature regarding possible diagnostic alternatives and organizes findings into 7 areas of study: demographic and medical history variables, seizure semiology, provocative testing, prolactin levels, single photon emission computed tomography, psychological testing, and neuropsychological testing. For each area, the literature is summarized, and conclusions about the accuracy of the technique as a diagnostic tool are drawn. Overall, it appears unlikely that any of the reviewed alternative techniques will replace video EEG monitoring; rather they may be more successful as complementary diagnostic tools. An important focus for further investigations involves combinations of diagnostic techniques for the differential diagnosis of epilepsy and PNES. PMID- 12090722 TI - Miltefosine Zentaris. AB - Zentaris (formerly Asta Medica) has developed and launched miltefosine, a phospholipid derivative, in various European countries [173324], [254947] and in Brazil in 1997 [332428], [366795] as a treatment for breast cancer. As of January 2002, phase III trials for treatment of leishmaniaisis were ongoing [435767]. In February 1999, Lehman Brothers predicted miltefosine would be launched for treatment of leishmaniasis in 2002, with peak sales expected in 2010 [319225]. PMID- 12090724 TI - Overview: interleukin-16 and its role in inflammatory bowel disease. PMID- 12090723 TI - Ravuconazole Eisai/Bristol-Myers Squibb. AB - Eisai and Bristol-Myers Squibb (BMS) are developing the triazole, ravuconazole, as a potential treatment for fungal infection [187888]. Eisai selected the compound for further development on the basis of its good safety profile and well balanced antifungal activity [187888]. Ravuconazole has a broader antifungal spectrum than fluconazole and itraconazole, particularly against strains of Candida krusei and Cryptococcus neoformans [271854], [342757], [370312]. By June 1999, the compound was undergoing phase II trials [327113]. In November 2001, it was reported that BMS was seeking a co-development partner for the compound [430011]. In October 2001, analysts at ABN Amro predicted sales of US $50 million in 2003 [444020]. PMID- 12090725 TI - Role of neurotrophins in inflammation of the gut. AB - Until now neurotrophins like nerve growth factor (NGF), brain-derived neurotrophic factor (BDNA), neurotrophin (NT)-3 and neurotrophic factors like glial-derived neurotrophic factor (GDNF) have been almost exclusively investigated concerning their role in differentiation, growth and survival of specific neurons in the peripheral and central nervous system. However, in the last decade several non-neuronal functions of neurotrophins and neurotrophic factors have been characterized. In the gastrointestinal tract, neurotrophins and neurotrophic factors regulate neuropeptide expression, interact with immunoregulatory cells and epithelial cells and regulate motility during inflammation. This highlights this new and complex regulatory system as important and may lead to new options in the treatment of acute and chronic inflammation of the gut. PMID- 12090726 TI - Angiotensin II as a pro-inflammatory mediator. AB - Angiotensin II (Ang II), the most important component of the renin-angiotensin system, is usually associated with hypertension and renal failure. Through its pro-inflammatory actions, it also plays an important role in each step of the development of atherosclerotic plaques and plaque rupture. Ang II stimulates the expression of nuclear factor-kappaB (NFkappaB), a transcription factor which regulates gene expression of inflammatory cytokines such as interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1). Ang II type 1 receptors (AT1) and angiotensin converting enzyme (ACE) are dramatically increased in atherosclerotic plaques, particularly in monocytes at the fibrous cap. Thus, in multiple ways, Ang II is a critical factor in atherosclerotic plaque formation, inflammation and plaque stability. ACE inhibitors and AT1R inhibitors could therefore be appropriate therapeutic agents in the treatment of atherosclerosis. PMID- 12090727 TI - Sampatrilat Shire. AB - Sampatrilat is a dual inhibitor of neutral endopeptidase (NEP) and angiotensin converting enzyme (ACE), which is under development by Pfizer and Shire (previously Roberts Pharmaceutical) for the potential treatment of hypertension. As of 1995, Pfizer had taken the compound into phase II clinical trials [179233]. In April 1997, Roberts Pharmaceutical, through its wholly owned subsidiary Roberts Laboratories Inc, received an exclusive license from Pfizer to develop and market sampatrilat for the treatment of essential hypertension and congestive heart failure. The agreement provided for transfer of data and the awarding of patent rights to Roberts [240809]. Although in June 2000, Shire confirmed it had discontinued all development of this drug [372652], by December 2000 it had recommenced studies with a reformulation of sampatrilat that achieved a 4-fold increase in oral bioavailability. A clinical trial with the new formulation was to be initiated in 2001, with results expected during the second quarter of 2001. The company also revealed that it was seeking a licensing partner at this time [394238]. In November 2001, Shire confirmed that while the project was undergoing further development with a view to outlicensing, it was unlikely to take this project into phase II alone [429562]. PMID- 12090728 TI - Treprostinil sodium Pharmacia. AB - United Therapeutics Corp (UTC) is developing treprostinil sodium (Remodulin, UT 15), a stable structural analog of prostacyclin, for the potential treatment of primary pulmonary (arterial) hypertension (PAH), peripheral vascular disease (PVD) and other cardiovascular conditions [327593], including critical limb ischemia (CLI) [412483]. In August 2000, UTC submitted the initial, non-clinical sections of an NDA for the treatment of pulmonary hypertension [378906]. Treprostinil, which had previously been designated as an Orphan Drug, was also awarded Priority Review status by the US FDA in October 2000 [385864], [386271]. In December 2000, UTC agreed with the FDA that the NDA for treprostinil did not need to be presented to the Cardiovascular and Renal Drugs Advisory Committee, which was expected to allow UTC and the FDA to work towards the 6-month Priority Review timeline [393888]. On August 9, 2001, the advisory committee recommended approval of treprostinil and UTC refiled the NDA on the same day [418682]. In February 2002, the FDA issued an approvable letter for treprostinil injection for the treatment of PAH. The FDA proposed drug labeling for PAH consistent with the treatment of both primary and secondary pulmonary hypertension in patients with New York Heart Association (NYHA) Class II-IV symptoms. The approvable letter also stated that the FDA intended to approve treprostinil with a requirement that UTC subsequently conduct a post-marketing controlled clinical trial to verify and further describe the drug's clinical benefit [439278]. In February 2001, UTC submitted a marketing authorization application (MAA) in France for approval of treprostinil for the treatment of PAH. Upon approval of the MAA, UTC planned to file for Mutual Recognition in other European countries and was also preparing similar submissions to non-European countries [391986], [397958]. By early 2001, phase II trials of treprostinil for the treatment of CLI were underway [412483]. In March 2001, the company was planning a phase III pivotal study in late-stage PVD by the end of 2001 [424180]. In April 2000, UTC was issued US-06054486 for the method of treating PVD with treprostinil [364130]. In February 2000, UTC entered into an agreement with Paladin Labs for the exclusive Canadian distribution of treprostinil for the remainder of clinical trials and after regulatory approvals [357302]. In November 2000, UTC and Antigen Pharmaceuticals entered into a strategic alliance for the distribution of treprostinil in the UK and Ireland [390157]. In November 2000, Deutsche Banc Alex Brown predicted a sales potential of US $250 million to US $350 million [418736]. In August 2001, Merril Lynch predicted sales of US $10 million to $20 million in 2002 [420652]. PMID- 12090729 TI - Overview. Cholecystokinin and eating. AB - This review focuses on the satiating effect of cholecystokinin (CCK) in humans. Current evidence supports a role for the peptide as a physiological satiety factor. The therapeutic potential of CCK analogs cannot be estimated until further studies are performed that demonstrate the efficacy of analogs for decreasing body weight, and the safety of CCK when administered chronically. Because the brain-gut axis is so important in the normal control of food intake, a great deal of caution is needed, particularly with agents that will presumably be taken for prolonged periods. Even so, at present it seems likely that we will see the introduction of CCK analogs or CCK antagonists as a new class of appetite modulating agents in the not-too-distant future. PMID- 12090730 TI - Tachykinin receptor antagonists in irritable bowel syndrome. AB - Tachykinins (TKs) are abundantly expressed in the gastrointestinal (GI) tract in intrinsic excitatory motor neurons, interneurons, sensory neurons and extrinsic sensory neurons. Their role in the regulation of enteric secretomotor functions is well established, especially following pathophysiological stimuli. Recent evidence emphasizes the role of TKs in the sensitization of peripheral branches of visceral afferent neurons, implying a role in determining visceral hypersensitivity. Furthermore, the involvement of both CNS and peripheral TK receptors in autonomic reactions to stress, render these receptors an appealing target for the development of drugs aimed at the treatment of irritable bowel syndrome (IBS), a functional GI disorder. The available preclinical evidence indicates that TK receptor antagonists could normalize motor disturbance (diarrhea and constipation) and reduce the painful symptoms that characterize IBS. PMID- 12090731 TI - Topiglan MacroChem. AB - MacroChem is developing Topiglan, a topical gel containing prostaglandin E1 (PGE1 or alprostadil) and its patented through-the-skin absorption enhancer excipient (SEPA), for the potential treatment of erectile dysfunction. By September 2000, it was in phase III clinical trials [382682]. By October 2000, MacroChem expected to file an NDA for Topiglan in late 2002 [387433]. In January 2002, at the JP Morgan Hambrecht & Quist 20th Annual Healthcare Conference in San Francisco, CA, MacroChem stated that further phase III studies were planned, including a reformulation that could reduce initation and increase dosage, as well as providing new packaging that improves dose consistency These studies should be underway by the third quarter of 2002, with a potential launch in late 2003 or early 2004 [436390]. The company's patented SEPA technology used in Topiglan is an absorption enhancer for transdermal delivery that has the potential to effectively increase the passage of therapeutic agents through the skin. The application of SEPA alone to the skin of carcinogenic-sensitive rats did not cause the development of tumors attributable to the SEPA [357621]. The company received a US notice of allowance of all patent claims covering Topiglan in February 1999. The patent (US-05942545) was issued in August 1999 [316028], [337720]. Further patent applications are pending in Canada, Europe and Japan [337720]. In February 2000, MacroChem filed further patent applications in 13 countries in Europe, Asia and Latin America and with the EPO [355947]. PMID- 12090732 TI - Vardenafil Bayer Yakuhin. AB - Bayer is developing vardenafil, an orally active phosphodiesterase (PDE) 5 inhibitor for the potential treatment of erectile dysfunction (ED) [314382]. NDAs were filed in September 2001 in the US and Mexico [423096], and vardenafil was submitted for Canadian approval in October 2001. As of November 2001, Bayer was expecting a response from the FDA in the second half of 2002 [429499]; the EMEA accepted a filing in January 2002, following a December 2001 submission [438163]. By October 2000, phase III trials were underway in Japan [384751] and by December 2001, a Japanese NDA had been filed; at the same time an application was filed in South Africa [426526], [433060]. At this time Japanese launch was expected in 2003 [434758]. By February 2001, Bayer was also investigating a nasal formulation of vardenafil for the potential treatment of erectile dysfunction [397608]. In November 2001, Bayer and GlaxoSmithKline signed a worldwide copromotion agreement for vardenafil, under which Bayer was to be responsible for all regulatory work required to obtain approval [429499]. In February 1999, Lehman Brothers predicted a 10% probability that vardenafil would reach the market, with launch in 2002. Peak Japanese sales of US$600 million were predicted for 2014 [319225]. In May 2000, Bayer predicted peak sales of Euro900 million [397137]. In July 2001, Lehman Brothers predicted a 75% chance that vardenafil would reach the market, and forecast peak sales of US $0.85 billion worldwide; the analyst also speculated that Bayer would seek a comarketing partner [414766]. PMID- 12090733 TI - Methylnaltrexone Progenics. AB - Progenics is developing methylnaltrexone (MNTX), an opioid antagonist licensed from UR Labs and the University of Chicago, for the potential treatment of the side effects of opioid pain therapy, such as constipation and post-operative bowel dysfunction. MNTX was discovered at the University of Chicago and subsequently licensed by UR Labs. In October 2001, Progenics entered into an agreement with UR Labs to obtain exclusive worldwide rights to the drug [423791]. MNTX is in phase II trials, with phase III studies expected to begin in 2002 [423791]. In October 2001, a double-blind, randomized, phase II study evaluating sc MNTX in cancer patients for the treatment of opioid-induced constipation was initiated at the University of Chicago Medical Center [423791]. In December 2001, the company stated that it was preparing to initiate phase IIb trials of the compound in opioid-induced constipation and post-operative bowel dysfunction [423791], [432507]. These trials were initiated in February 2002 [440995]. PMID- 12090734 TI - Dexloxiglumide Rotta Research Lab. AB - Dexloxiglumide, the (R)-isomer of loxiglumide, is a selective and highly potent CCK1 receptor antagonist. It is twice as potent as the racemic compound. because the anti-CCK activity is specific to the (R)-form, whereas the (S)-isomer is almost ineffective. It has been developed by Rotta Research Lab SpA for the treatment of diseases in which CCK1 receptor activity is potentially involved, including gastrointestinal motility, food intake and pancreatic disorders [218696]. Its receptor-mediated actions have been described in multiple in vitro and in vivo pharmacological systems. Results from both preclinical and clinical studies indicate that it is an effective inhibitor of gallbladder contraction, improves lower esophegal sphincter (LES) function, accelerates gastric emptying, accelerates colonic transit and significantly decreases symptoms in IBS and functional dyspepsia patients, and therefore has potential as an effective treatment for constipation-predominant IBS. functional dyspesia, constipation, LES function, gastric emptying disorders and biliary colics. Forest Laboratories has entered into an agreement with Rotta for the development and marketing of dexloxiglumide for the treatment of constipation-predominant IBS and phase III studies are currently ongoing in the US. In August 2000, Merrill Lynch expected that dexloxiglumide would not be launched until 2004 [379892], and in June 2001, predicted a US filing date in 2003 [413928]. PMID- 12090735 TI - Review of the clinical experience with 5-azacytidine and 5-aza-2'-deoxycytidine in solid tumors. AB - In recent years the importance of epigenetic changes in carcinogenesis has been unfolding. It is now clear that the fifth base of the genome, methylcytosine, plays a critical role in the control of gene expression during normal development and carcinogenesis. Efforts to decrease methylation in neoplasias as a therapeutic strategy have been productive in hematologic malignancies but disappointing in solid tumors. The following is a review of the clinical experience with the agents 5-azacytidine and 5-aza-2'-deoxycytidine in solid malignancies and a discussion of the difficulties encountered. PMID- 12090736 TI - Chemosensitization of solid tumors by modulation of resistance mechanisms. AB - The number of drugs available for chemotherapy is growing exponentially, and this trend looks set to continue. Chemosensitization strategies use the administration of one drug or agent to render cancer cells more susceptible to a second agent. Modulation of resistance mechanisms due to xenobiotic membrane pumps such as the multidrug resistant proteins, MDR1/P-glycoprotein or MRP is feasible and a number of new agents have been produced to inhibit drug efflux resulting from expression of these molecules. However, tumor cells may express or upregulate more than one such molecule at one time, and this approach is unlikely to benefit every patient. Detoxification mechanisms mediated by glutathione conjugation or metallothionein are also responsible for resistance--the former has been linked to MRP-mediated resistance. Again, modulation is possible but may increase the toxicity of drugs to normal tissues and an increased therapeutic index is not guaranteed. Tumors exposed to DNA damaging agents often upregulate DNA repair mechanisms and this contributes to resistance. Different pathways perform the repair of different forms of DNA damage, and it is difficult to inhibit all of these. Nevertheless, inhibition of DNA repair can re-sensitize tumors to chemotherapy and is increasingly exploited. One of the most successful and widely used approaches is to combine gemcitabine with an alkylating or platinating agent. While gemcitabine may inhibit DNA polymerases directly, this cytidine analog is also likely to be incorporated by DNA repair leading to activity against non-cycling cells, which form the majority of the neoplastic cell population in most solid tumors. Oncologists should take account of potential resistance mechanisms when treating patients: it is often feasible to design combinations with old or new drugs which exploit these apparent weaknesses to the patient's advantage. PMID- 12090737 TI - Chemosensitization of solid tumor cells by alteration of their susceptibility to apoptosis. AB - Chemosensitization strategies use the administration of one drug or agent to render cancer cells more susceptible to a second agent. Usually this involves enhanced drug metabolism, improvement of drug uptake or blockage of resistance mechanisms. Alteration of the susceptibility of cancer cells to apoptosis, the process of individual cell death by which many chemotherapeutic drugs act, shows particular promise for therapy in the future, and is the focus of this review. The dependence of cancer cells on non-neoplastic cells to form solid tumors allows anti-angiogenic therapy to be used in conjunction with chemotherapy to increase the therapeutic index. Chemosensitization strategies are set to become increasingly important in cancer therapy, allowing rational design of synergistic drug combinations at an earlier stage in drug development. PMID- 12090738 TI - MitoExtra SuperGen. AB - SuperGen Inc is developing a cyclodextrin-based reformulation of mitomycin C (MitoExtra) for the potential treatment of gastic, pancreatic, breast lung and colorectal cancers. In April 2002, the NDA was pending approval from the US FDA after a March 2002 filing of a final response including data from a phase II trial [447618]. MitoExtra is a proprietary reformulation of the approved anticancer drug mitomycin C. SuperGen's reformulation is based on technology designed to improve the handling characteristics and safety profile of mitomycin C and other anticancer drugs by enhancing the drug's stability in solution form and 'shielding' it at the injection site [205482]. In a phase II study in patients with advanced solid tumors, MitoExtra showed no evidence of diminished efficacy or unexpected toxicity [447618]. In April 2000, SuperGen' was issued US 06048845, protecting the company's proprietary Extra technology platform [363494]. PMID- 12090739 TI - Rates of progression to hypertension among non-hypertensive subjects: implications for blood pressure screening. PMID- 12090740 TI - The Gly389Arg beta-1 adrenoceptor polymorphism and cardiovascular disease: time for a rethink in the funding of genetic studies? PMID- 12090741 TI - Acute coronary syndrome: the struggle for the best in risk stratification and therapy. PMID- 12090742 TI - Reperfusion damage or no-reflow damage in primary coronary interventions in acute myocardial infarction? PMID- 12090743 TI - Sympathetic activation and malignant ventricular arrhythmias: a molecular link? PMID- 12090744 TI - Exercise-related syncope: are athletes different from sedentary subjects? PMID- 12090745 TI - Is genotype or phenotype the better tool for investigating the role of ACE in human cardiovascular disease? PMID- 12090746 TI - An evaluation of the beta-1 adrenergic receptor Arg389Gly polymorphism in individuals at risk of coronary events. A WOSCOPS substudy. AB - AIMS: The Glycine389 variant of the beta-1 adrenergic receptor generates markedly less cAMP when stimulated in vitro than the more prevalent Arginine389 variant and may confer protection against coronary events similar to that observed with beta-blockers. The aim of this study was to ascertain whether this Glycine389 variant protects against coronary events. METHODS AND RESULTS: We identified the genotype at position 389 of the beta1AR in 1554 individuals taken from men enrolled in the West of Scotland Coronary Prevention Study. Men with a coronary event (event group) were each matched for age and smoking status with two control subjects from the same cohort who had not had a coronary event (control group). We compared the distribution of genotypes in the event and control groups. Conditional logistic regression was used to calculate odds ratios for each of the genotypes. The prevalence of the three genotypes in the entire cohort was ArgArg 53.5%, ArgGly 39.6%, GlyGly 6.9%. The Arg389Gly beta-1 adrenergic receptor polymorphism was not associated with coronary events. Using the ArgArg genotype as the reference, the odds ratio for the ArgGly genotype was 1.1 (95% CI, 0.88 1.38) and for the GlyGly genotype it was 1.05 (95% CI, 0.68-1.62). CONCLUSION: Our longitudinal case-control study demonstrates that the Glycine389 variant of the beta-1 adrenergic receptor does not protect against coronary events. PMID- 12090747 TI - Continuous multilead ST-monitoring identifies patients with unstable coronary artery disease who benefit from extended antithrombotic treatment. AB - AIMS: Prolongation of anticoagulant treatment might reduce subsequent cardiac events in patients with unstable coronary artery disease. Multilead ST-segment monitoring identifies patients with a high risk of adverse outcome. The aim was to assess the value of multilead ST-monitoring in prospectively identifying patients who respond to extended anticoagulant treatment with low-molecular weight heparin when treated by a primarily non-invasive strategy. METHODS AND RESULTS: In this substudy of the FRISC II trial, ST-monitoring with a continuous 12-lead ECG or vectorcardiography was performed for 24 h in 629 patients with unstable coronary artery disease randomized to receive either the low-molecular weight heparin dalteparin, or placebo for 3 months after at least 5 days' dalteparin treatment in all patients. Ischaemic episodes were detected in 34% during ST-monitoring. In the group with ischaemic episodes, the extended dalteparin treatment was associated with a lower rate of death, myocardial infarction, or revascularization (35.2% vs 53.4%, relative risk reduction: 34%, P=0.01). In patients without ischaemic episodes, long-term dalteparin treatment had no effect. CONCLUSIONS: In patients with unstable coronary artery disease treated primarily with a non-invasive strategy, ischaemic episodes revealed while on multilead ST-monitoring identifies patients who benefit most from extended treatment with anticoagulants. PMID- 12090748 TI - Does eptifibatide confer a greater benefit to patients with unstable angina than with non-ST segment elevation myocardial infarction? Insights from the PURSUIT Trial. AB - AIMS: To evaluate the differential effects of eptifibatide therapy on unstable angina vs non-ST elevation myocardial infarction at enrollment, since the separate impact on these two major diagnostic subsets of acute coronary syndrome patients has not been fully investigated. METHODS AND RESULTS: We examined the 9461 patients in the PURSUIT trial (conducted between 1995 and 1997) to compare the effects of eptifibatide on unstable angina and myocardial infarction. The study showed greater and more consistent effects of eptifibatide therapy on unstable angina than non-ST elevation myocardial infarction in reducing 30-day death/(re)infarction (from the unadjusted rate of 13.0% to 11.2%, P=0.059 for unstable angina; and 18.9% to 17.9%, P=0.387 for myocardial infarction), especially among patients who underwent early percutaneous coronary intervention (odds ratios=0.49 and 0.86, 95% confidence intervals=0.30-0.80 and 0.53-1.42, respectively, for unstable angina and myocardial infarction). The only subgroup for whom the benefit of eptifibatide was not evident was female myocardial infarction patients who did not undergo early percutaneous coronary intervention. CONCLUSIONS: These data suggest that eptifibatide benefited unstable angina patients more than myocardial infarction patients, especially among those who underwent early percutaneous coronary intervention, and support its use as concomitant therapy with early percutaneous coronary intervention especially in female myocardial infarction patients. PMID- 12090749 TI - Incidence and clinical significance of distal embolization during primary angioplasty for acute myocardial infarction. AB - AIMS: Although recognized as an important feature of atherosclerotic coronary disease, little is known about the frequency and prognostic importance of distal embolization during primary angioplasty for acute myocardial infarction. METHODS AND RESULTS: As part of a randomized trial of thrombolysis vs primary angioplasty, 178 patients with acute myocardial infarction were treated with primary angioplasty. In these patients the occurrence of distal embolization after angioplasty was assessed. Embolization was defined as a distal filling defect with an abrupt 'cutoff' in one of the peripheral coronary artery branches of the infarct-related vessel, distal to the site of angioplasty. We analysed myocardial blush grade, ST-T segment elevation resolution, enzymatic infarct size and left ventricular ejection fraction in patients with and without distal embolization. Clinical information was collected for a mean of 5 years. Distal embolization was present in 27 patients (15.2%). Mean age and gender were not different from patients without distal embolization. Angiographic success (thrombolyis in myocardial infarction flow grade 3 and residual stenosis <50%) after primary angioplasty was less frequently observed in patients with distal embolization (70% vs 90%, P<0.01). Myocardial blush and ST-T segment elevation resolution after angioplasty were reduced when distal embolization was present. Patients with distal embolization had a larger enzymatic infarct size (mean cumulative lactate dehydrogenase measured over 72 h, 1612 vs 847, P<0.05) and a lower left ventricle ejection fraction at discharge (42% vs 51%, P<0.01). Long term mortality was higher in patients with distal embolization (44% vs 9%, P<0.001). CONCLUSION: Distal embolization in patients treated with primary angioplasty is visible on the coronary angiogram in 15.2% of patients. It is related to reduced myocardial reperfusion, more extensive myocardial damage and a poor prognosis. Additional pharmacological interventions and/ or mechanical devices should be studied to prevent and/or treat distal embolization. PMID- 12090750 TI - Importance of sympathetic activation for the expression of Na+-Ca2+ exchanger in end-stage failing human myocardium. AB - AIMS: In end-stage heart failure, an alteration in the expression of the Na+-Ca2+ exchanger has been reported. Regulation of its expression is largely unknown. We sought to find out whether Na+-Ca2+ exchanger in human heart failure is regulated by sympathetic activation. In addition, since Na+-Ca2+-exchange is electrogenic, we conjectured whether increased expression of Na+-Ca2+ exchanger is associated with an increased incidence of cardiac arrhythmias. METHODS AND RESULTS: Twenty three patients suffering from end-stage cardiac failure were examined in the hours preceding cardiac transplantation. Plasma levels of norepinephrine, epinephrine, atrial natriuretic peptide, renin activity, aldosterone, tumour necrosis factor (TNF)-alpha, and TNF-receptors were measured. All parameters were elevated relative to 21 healthy control subjects. As determined by immunoblots, protein levels of the Na+-Ca2+ exchanger were increased by 56% and protein levels of sarcoplasmic reticulum (SR) Ca2+-ATPase were decreased by 20% in left ventricles of the explanted failing hearts. A significant correlation between protein and neurohumoral levels was exclusively found for the Na+-Ca2+ exchanger with norepinephrine (r=0.64; P=0.01). Recent Holter ECGs revealed that patients with sustained or non-sustained ventricular tachycardia (more than three consecutive beats) had significantly higher Na+-Ca2+ exchanger protein and plasma norepinephrine levels than patients with a maximum of two consecutive beats (Na+ Ca2+ exchanger: 109+/-10 vs 83+/-7, n=11 each, P<0.05; norepinephrine: 1359+/-159 vs 656+/-88 pg. ml(-1), n=9 each, P<0.001). CONCLUSIONS: Sympathetic activation may enhance the expression of Na+-Ca2+ exchanger in end-stage heart failure. The data support the hypothesis that increased Na+-Ca2+-exchange could favour malignant ventricular arrhythmias. PMID- 12090751 TI - Exercise-related syncope in young competitive athletes without evidence of structural heart disease. Clinical presentation and long-term outcome. AB - AIMS: Exercise-related syncopal spells in athletes receive great attention and are a source of anxiety in the sporting world. The aim of the present study is to describe the clinical presentation, the yield of the initial diagnostic work-up and the long-term outcome of a series of consecutive competitive athletes with recurrent exercise-related syncopal spells. METHODS AND RESULTS: The study cohort included 33 athletes (20 females, mean age 21.4+/-3.2 years) referred for recurrent unexplained episodes of exercise-related syncope (mean number of spells before evaluation 4.66+/-1.97). All athletes underwent an extensive evaluation, including echocardiography, 24-h electrocardiographic monitoring, exercise testing, cardiac electrophysiological study and head-up tilt testing. The echocardiographic examination revealed the presence of a mitral valve prolapse in two cases (6.0%). During maximal exercise testing, four athletes (12.1%) developed hypotension associated with pre-syncope. Twenty-two subjects (66.6%) showed a positive response to head-up tilt testing. During follow-up (33.5+/-17.2 months) 11/33 athletes (33.3%) showed at least one recurrence of exercise-related syncope (mean time to first recurrence 20.4+/-14.5 months). No other adverse event of any kind was noted during follow-up. The Kaplan-Meier estimates of first recurrence of exercise-related syncope after 12, 36 and 60 months were 9.1%, 24.4% and 42.9%. The number and frequency of exercise-related syncopal spells before evaluation were found to be univariate predictors of syncope recurrence (P<0.001). However, in the multivariate analysis, the number of exercise-related syncopal spells before evaluation was found to be the only independent predictor of syncope recurrence (P<0.05). CONCLUSIONS: These findings support the idea that recurrent exercise related-syncope is not associated with an adverse outcome in athletes without cardiac disease. PMID- 12090752 TI - Ventricular mapping during atrial and ventricular pacing. Relationship of multipotential electrograms to ventricular tachycardia reentry circuits after myocardial infarction. AB - AIMS: Conduction through separated myocyte bundles causes multipotential electrograms and reentrant ventricular tachycardia. We hypothesized that without initiating tachycardia, the reentry region could be detected by analysing the change in multipotential electrograms during two different activation sequences. METHODS AND RESULTS: During catheter mapping and ablation in 16 patients with ventricular tachycardia late after infarction ventricular electrograms were recorded from 1072 sites during atrial and right ventricular paced ventricular activation. Multipotential electrograms were present during both activation sequences at 285 (27%) sites, during atrial pacing only at 159 (15%) sites and during right ventricular pacing only at 152 (14%) sites. Sites with multipotential electrograms during both activation sequences were more often related to a ventricular tachycardia circuit isthmus (43%) as compared to sites where such electrograms were present during one activation sequence (20%). Multipotential electrograms with >2 low amplitude deflections and a >100 ms difference in duration between the two activation sequences were infrequent but highly predictive of the reentry circuit. CONCLUSION: Regions with fixed multipotentials consistent with conduction block might be useful guides for ablation approaches that target large regions of the infarct, but are not sufficiently specific to be the sole guide for focal ablation approaches. PMID- 12090755 TI - Mobilization of dendritic cells from patients with breast cancer into peripheral blood stem cell leukapheresis samples using Flt-3-Ligand and G-CSF or GM-CSF. AB - Treatment with myeloablative chemotherapy and autologous peripheral blood stem cell (PBSC) transplantation followed by vaccination with autologous dendritic cells (DCs) treated with tumor antigens is a promising therapeutic strategy for several types of cancer. Obtaining sufficient numbers of both PBSCs and DCs is central to this approach. Previously, it has been shown that administration of Flt-3-Ligand (FL) combined with either G-CSF or GM-CSF mobilizes large numbers of PBSCs in patients with cancer. In the current study, we sought to determine whether these same cytokines could simultaneously mobilize DCs into the PBSC leukapheresis collection. DCs were analysed in PBSC leukapheresis samples obtained from five patients with high-risk breast cancer who received G-CSF alone as priming prior to leukapheresis, four patients who received FL+G-CSF and five patients who received FL+GM-CSF. DCs were defined as cells with a lin(dim/-) HLA DR+ CD11c+ phenotype. The proportions of DCs in the FL+G-CSF and FL+GM-CSF samples were significantly higher than in pre-mobilization peripheral blood and G CSF leukapheresis samples. The mean yield of DCs/kg in the FL+GM-CSF samples was also significantly higher than the mean yield of DCs in the G-CSF samples. The FL+G-CSF and FL+GM-CSF mobilized DCs were immature by morphologic and phenotypic criteria but stimulated allogeneic T-cells at levels similar to DCs generated in culture from PBMCs. Overnight culture?of the immature DCs obtained from patients receiving either FL+G-CSF or FL+GM-CSF in TNF-alpha?resulted in the generation of mature DCs. In summary, administration of FL in combination with GM-CSF and G-CSF to patients with breast cancer can mobilize large numbers of immature DCs into PBSC leukapheresis collections. PMID- 12090754 TI - Cardiotrophin-like cytokine induces astrocyte differentiation of fetal neuroepithelial cells via activation of STAT3. AB - Cardiotrophin-like cytokine (CLC), also known as novel neurotrophin-1/B cell stimulating factor-3 (NNT-1/BSF-3), is a recently identified member of the interleukin (IL)-6 family of cytokines that share gp130 as a signal-transducing receptor component. In this study, we demonstrate that CLC is expressed in fetal mouse neuroepithelial cells and has a potential to induce their astrocyte differentiation in a synergistic manner with bone-morphogenetic protein (BMP)-2, which is also expressed in the fetal mouse brain. CLC-stimulation led to promoter activation of the gene for an astrocyte marker, glial fibrillary acidic protein (GFAP), which was clearly inhibited by expression of a dominant negative form of a transcription factor, STAT3, or by introduction of a mutation in a single STAT3 binding site in the promoter, suggesting a critical role of STAT3 in the CLC induced GFAP transcription. These results suggest that CLC plays a role in astrocyte differentiation via STAT3 activation within the developing brain. PMID- 12090756 TI - DNA microarray analysis of transforming growth factor-beta and related transcripts in nasal biopsies from patients with allergic rhinitis. AB - Decreased activity of anti-inflammatory cytokines like transforming growth factor (TGF)-beta may contribute to allergic inflammation. In vivo effects of TGF-beta effects are difficult to infer from local concentrations, since TGF-beta-effects depend on a complex system of regulatory proteins and receptors. Instead the effects of TGF-beta might be inferred by examining TGF-beta-inducible transcripts. In this study DNA microarrays were used to examine local expression of TGF-beta, TGF-beta-regulatory and -inducible transcripts in nasal biopsies from patients with symptomatic allergic rhinitis and healthy controls. In addition, nasal fluids were analysed with cytological and immunological methods. Nasal fluid eosinophils, albumin, eosinophil granulae proteins and IgE, but not TGF-beta, were higher in patients than in controls. DNA microarray analysis of nasal mucosa showed expression of transcripts encoding TGF-beta, TGF-beta regulatory proteins and -receptors at variable levels in patients and controls. By comparison, analysis of 28 TGF-beta-inducible transcripts indicated that 23 of these had lower measurement values in patients than in controls, while one was higher, and the remaining four were absent in both patients and controls. In summary, TGF-beta and a complex system of regulatory genes and receptors are expressed in the nasal mucosa. Low expression of TGF-beta-inducible transcripts may indicate decreased TGF-beta activity in allergic rhinitis. DNA microarray analysis may be a way to study cytokine effects in vivo. PMID- 12090757 TI - Oncostatin M induces tissue-type plasminogen activator and plasminogen activator inhibitor-1 in Calu-1 lung carcinoma cells. AB - Oncostatin M (OSM) is a glycoprotein cytokine that is produced by activated T lymphocytes, monocytes, and macrophages. In a DNA synthesis assay, OSM reduced tritiated thymidine incorporation by 53% in Calu-1 lung carcinoma cells. Radiolabeled cDNAs from untreated Calu-1 cells and 30-h OSM-treated cells were used to probe duplicate nylon membrane cDNA expression arrays. This study revealed OSM-mediated expression of mRNAs encoding tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1). Northern blot analysis showed that the steady-state level of tPA mRNA is nearly undetectable in Calu-1 cells. Exposure of these cells to OSM for 30 h increased tPA mRNA expression by 20-fold and PAI-1 mRNA expression by 5-fold. Exposure of these cells to other gp130 receptor family cytokines, including leukemia inhibitory factor (LIF), interleukin-6 (IL-6), and IL-11, do not significantly affect DNA synthesis or induction of tPA/PAI-1. Western blot studies demonstrated that OSM mediates a marked increase in secretion of the tPA protein. Secreted tPA was present in the conditioned medium almost exclusively as tPA/PAI-1 complexes. Inhibitor studies demonstrated that OSM-mediated induction of tPA and PAI-1 mRNAs is largely dependent upon activation of the MEK1/2 pathway. The JAK3/STAT3 pathway potentially serves a secondary role in these regulatory events. PMID- 12090758 TI - Vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide inhibit LPS-stimulated MIP-1alpha production and mRNA expression. AB - Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) are neuropeptides with immunomodulatory properties, including the regulation of several proinflammatory mediators. Such mediators, for example chemokines, influence trafficking of inflammatory cells and contribute to shaping the immune response. In the present work, we studied the effect of VIP and PACAP on the CC chemokine macrophage inflammatory protein-1 alpha (MIP-1alpha) production in LPS-stimulated RAW 264.7 macrophage cell line. VIP and PACAP inhibited the production of MIP-1alpha in a dose-dependent manner and over a broad spectrum of LPS concentrations. The use of selective agonists and antagonists of VIP/PACAP receptors showed that type 1 VIP receptor (VPAC1) is the major receptor involved, but the type 2 VIP receptor (VPAC2) may be also implicated. By using selective PKA and PKC inhibitors and cAMP mimicked agents, we demonstrated a cAMP-dependent signalling pathway for the inhibitory effect of VIP/PACAP on MIP-1alpha production, although a minor non-mediated cAMP pathway was also involved. mRNA expression studies showed a down-regulation of MIP-1alpha gene expression by VIP and PACAP. Taken together, the present work strongly supports an anti-inflammatory role of VIP and PACAP by a new mechanism associated with impairment of a key component of the chemokine network. PMID- 12090759 TI - Endocrine expression of the active form of TGF-beta1 in the TGF-beta1 null mice fails to ameliorate lethal phenotype. AB - TGF-beta1 null mice die by 3 to 4 weeks of age due to a severe autoimmune mediated multifocal inflammation resulting in multi-organ failure. To assess the therapeutic potential of circulating levels of active TGF-beta1, we generated mice with endocrine expression of active TGF-beta1 on a TGF-beta1 null background (TGF-beta1 (-/-/TG)) by crossing TGF-beta1(+/-) mice with transgenic mice (TG) that express recombinant TGF-beta1 specifically in the liver and secrete it in the blood. The TGF-beta1 (-/-/TG) mice exhibit a survival profile similar to the TGF-beta1 (-/-) mice indicating a failure to rescue the lethal phenotype. However, serum TGF-beta1 levels in the TGF-beta1 (-/-/TG) mice were restored to near normal levels with expression in all the tissues, notably in the kidney and spleen. Histopathology showed reduced inflammation in the target tissues, especially in the heart. Interestingly, unlike TGF-beta1 (-/-) mice, the TGF beta1 (-/-/TG) mice have glomerulonephritis in their kidneys similar to the TG mice. Thus, the phenotype of TGF-beta1 (-/-/TG) animal model indicates the potential role of circulating active-TGF-beta1 in reducing inflammation, but its failure to rescue lethality in TGF-beta1 null mice indicates a critical autocrine role of TGF-beta1. PMID- 12090760 TI - Antagonism between interleukin 3 and erythropoietin in mice with azidothymidine induced anemia and in bone marrow endothelial cells. AB - Azidothymidine (AZT)-induced anemia in mice can be reversed by the administration of IGF-IL-3 (fusion protein of insulin-like growth factor II (IGF II) and interleukin 3). Although interleukin 3 (IL-3) and erythropoietin (EPO) are known to act synergistically on hematopoietic cell proliferation in vitro, injection of IGF-IL-3 and EPO in AZT-treated mice resulted in a reduction of red cells and an increase of plasma EPO levels as compared to animals treated with IGF-IL-3 or EPO alone. We tested the hypothesis that the antagonistic effect of IL-3 and EPO on erythroid cells may be mediated by endothelial cells. Bovine liver erythroid cells were cultured on monolayers of human bone marrow endothelial cells previously treated with EPO and IGF-IL-3. There was a significant reduction of thymidine incorporation into both erythroid and endothelial cells in cultures pre treated with IGF-IL-3 and EPO. Endothelial cell culture supernatants separated by ultrafiltration and ultracentrifugation from cells treated with EPO and IL-3 significantly reduced thymidine incorporation into erythroid cells as compared to identical fractions obtained from the media of cells cultured with EPO alone. These results suggest that endothelial cells treated simultaneously with EPO and IL-3 have a negative effect on erythroid cell production. PMID- 12090761 TI - Body fat: is it a handicap to racehorses? PMID- 12090763 TI - Rendering unto Caesar: welfare problems in Belgian Blue cattle. PMID- 12090762 TI - Bovine virus diarrhoea virus--which way to turn? PMID- 12090764 TI - Animal models for orthopaedic disease--who benefits? PMID- 12090765 TI - The sheep as a model for osteoporosis in humans. AB - There is great interest in large animal models for studying different aspects of osteoporosis. Several laboratories around the world have used ovariectomized sheep as a model because of their ease of housing and handling, low expense compared to other large animals, availability and acceptance in society as a research animal. They have been used to study the response to new therapies for post-menopausal osteoporosis, low-magnitude mechanical stimulation, orthopedic implants in osteoporotic bone and bioactive ceramics to strengthen vertebral bodies. To produce severely osteopenic bone comparable to that seen in humans, a combination of estrogen deficiency following ovariectomy plus a calcium-wasting diet is currently being investigated. PMID- 12090766 TI - Biomechanical analysis of the canine hind limb: calculation of forces during three-legged stance. AB - This paper presents a three-dimensional biomechanical model of the canine hind limb, and describes the process of determining the muscle forces and joint reaction forces and moments occurring in the hind limb during three-legged stance. The model was based on anatomical and morphometric data presented in a previous paper. Equations of equilibrium were formulated for the different components of the hind limb. Since the number of unknowns exceeded the number of equations, the problem was statically indeterminate. Two optimization techniques were applied to solve this statically indeterminate problem. The resultant hip joint reaction force (acting on the acetabulum) predicted by these optimization methods ranged between 0.73 and 1.04 times body weight, and was directed dorsally, medially and caudally. The resultant knee-joint reaction force (acting on the femur) ranged between 1.05 and 1.08 times body weight, and was directed dorsally, laterally and cranially. The largest muscle forces predicted by the minimization of maximal muscle stress (MMMS) criterion were in the biceps femoris (0.24 times body weight), rectus femoris (0.15 times body weight), medial gluteal (0.18 times body weight), semi-membranosus (0.09 times body weight), the lateral and intermediate vastus (0.18 times body weight) and the medial vastus (0.17 times body weight). The largest muscle forces predicted by the minimization of the sum of muscle forces (MSMF) criterion were in the biceps femoris (0.29 times body weight), lateral and intermediate vastus (0.45 times body weight)), and the deep gluteal (0.16 times body weight). The magnitudes and directions of the forces in the joints of the canine hind limb, as well as in the muscles that surround these joints, provide a database needed for future biomechanical analyses of the physiology and pathophysiology of the canine hind limb. PMID- 12090767 TI - Failure to spread bovine virus diarrhoea virus infection from primarily infected calves despite concurrent infection with bovine coronavirus. AB - Previous reports on the spread of bovine virus diarrhoea virus (BVDV) from animals primarily infected with the agent are contradictory. In this study, the possibility of transmission of BVDV from calves simultaneously subjected to acute BVDV and bovine coronavirus (BCV) infection was investigated. Ten calves were inoculated intranasally with BVDV Type 1. Each of the 10 calves was then randomly allocated to one of two groups. In each group there were four additional calves, resulting in five infected and four susceptible calves per group. Virulent BCV was actively introduced in one of the groups by means of a transmitter calf. Two calves, susceptible to both BVDV and BCV, were kept in a separate group, as controls. All ten calves actively inoculated with BVDV became infected as shown by seroconversions, and six of them also shed the virus in nasal secretions. However, none of the other eight calves in the two groups (four in each) seroconverted to this agent. In contrast, it proved impossible to prevent the spread of BCV infection between the experimental groups and consequently all 20 study calves became infected with the virus. Following infection, BCV was detected in nasal secretions and in faeces of the calves and, after three weeks in the study, all had seroconverted to this virus. All calves, including the controls, showed at least one of the following clinical signs during days 3-15 after the trial started: fever (> or =40 degrees C), depressed general condition, diarrhoea, and cough. The study showed that BVDV primarily infected cattle, even when co-infected with an enteric and respiratory pathogen, are inefficient transmitters of BVDV. This finding supports the principle of the Scandinavian BVDV control programmes that elimination of BVDV infection from cattle populations can be achieved by identifying and removing persistently infected (PI) animals, i.e. that long-term circulation of the virus without the presence of PI animals is highly unlikely. PMID- 12090768 TI - Fat-free mass is related to one-mile race performance in elite standardbred horses. AB - This study examined whether body composition was predictive of competitive success in elite standardbreds (STB). Rump fat and muscle thickness (MTH) (vastus lateralis/intermedius [VL], extensor carpi radialis [ECR]) were measured in vivo in male n=6; female n=8 by B-mode ultrasound. Percentage body fat (%fat) was calculated from rump fat. There were no gender differences for age, body mass (males 432+/-11 kg; females 443+/-13 kg), fat-free mass (FFM) (males 400+/-12 kg; females 400+/-11 kg), ECR MTH (males 61+/-2 cm; females 60+/-2 cm) or race time (RT) (males 113+/-3 s; females 114+/-2 s). Males had less (P<0.05) fat mass (males 32+4 kg; females 44+/-3 kg) and %fat (males 7.4+/-0.9%; females 9.9+/ 0.5%) and larger (P<0.05) VL MTH (males 88+/-7 cm; females 81+/-3 cm). RT was correlated to %fat and fat mass in males (r=0.89; r=0.82, P<0.05) not females (r=0.51; r=0.14). FFM tended to relate to RT in males (r=-0.76, P=0.07) and females (r=-0.59, P=0.12). Combined %fat and FFM data were correlated to RT (%fat r=0.70, P<0.01; FFM r=-0.65, P<0.01). RT was not correlated to MTH (VL r=-0.28; ECR r=-0.31). In conclusion, FFM was related to RT in elite STB with %fat negatively related to RT in males. PMID- 12090769 TI - Optimal timing of elective caesarean section in Belgian White and Blue breed of cattle: the calf's point of view. AB - The purpose of this study was to evaluate whether the timing of elective caesarean section (ECS) during parturition affects pulmonary and metabolic adaptation to extra-uterine life in healthy Belgian White and Blue (BWB) calves delivered at term. Vaginal palpation was performed and deliveries divided into six categories of timing for ECS: cervix closed (TECS 1); passive and active cervical dilatation (TECS 2 and TECS 3); full cervical dilatation (TECS 4); spontaneous rupture of allantoic (TECS 5) and amniotic (TECS 6) membranes. One hundred and eighteen BWB calves were examined at birth, 5, 15, 30, 45 and 60 min and 2, 3, 6, 12 and 24 h after birth using the following measurements: physical examination (time between birth and sternal recumbency [T-SR]); heart rate (mHR); arterial blood gas analyses (arterial partial pressure in oxygen [PaO(2)], in carbon dioxide [PaCO(2)], arterial haemoglobin oxygen saturation [SaO(2)], alveolo-arterial difference in oxygen [AaDO(2)]); pulmonary function tests using the oesophageal balloon catheter technique (respiratory rate [RR], total pulmonary resistance [R(L)], dynamic lung compliance [C(Ldyn)], tidal volume [V(T)] and minute volume [V(E)]); arterial and venous blood acid-base balance analyses (arterial and venous pH [pHa and pHv], bicarbonate concentration [HCO(3)a and HCO(3)v], base excess [BEa and BEv]); rectal temperature (RT); jugular venous blood sampling for determination of metabolic variables (blood glucose [G], plasma lactate [L], serum cortisol [C], plasma noradrenaline [NA] and adrenaline [A] concentrations); haematological variables (red blood cell count [RBC], total haemoglobin concentration [Hb], Packed Cell Volume [PCV]) and passive immune transfer variables (total serum protein [TP] and beta(2)gammaglobulin [beta(2)gamma] concentrations).TECS significantly (P or =4 calves, TECS< or =3 calves showed lower PaO(2), SaO(2), V(T), C(Ldyn), RT, G, NA, A, RBC, Hb and TP and higher AaDO(2), RR, V(E). TECS differences progressively decreased and disappeared between 6 and 12 h after birth in TECS 2 and 3 calves but remained up to 24 h in TECS 1 calves. Improved postnatal respiratory and metabolic adaptation in TECS> or =4 calves were mainly related to differences in exposure to labour and subsequent hormonal surge: catecholamines, particularly A, enabled more effective removal of lung liquid and/or release of surfactant which contribute to better gas exchanges and induced greater energy mobilization to maintain adequate body temperature. It was concluded waiting for full cervical dilatation before performing CS should be encouraged because it promotes postnatal respiratory and metabolic adaptation in full-term BWB calves. PMID- 12090770 TI - Plasma trypsin level in horses suffering from acute intestinal obstruction. AB - Gastrointestinal disorders in horses leading to endotoxic shock could have further consequences on other splanchnic organs such as the pancreas, as can be seen in humans suffering from septic shock. In this study, the range of enzymatically active trypsin (EAT) in healthy horses was established and is similar to the range observed in healthy humans. EAT values were determined in horses with acute abdominal crises on admission as well as during anaesthesia and in the postoperative phase. A significant increase in plasma EAT was found in 59% of the horses with surgical colic when compared to our established reference range. Significantly higher values were found in severe shock cases. When separated in groups according to the duration of colic before referral, significantly higher EAT values were observed in the non-survivor group compared to the survivor group of colics of short duration. EAT plasma values increased significantly during the postoperative phase, and were significantly higher in small intestine obstructions than in large bowel disorders. In human medicine, hypovolaemic or septic shock patients show an increase in pancreatic proteases. Splanchnic hypoperfusion during shock could lead to pancreatic damage resulting in trypsin liberation into the peritoneal space and an increase in plasma levels. Trypsin is able to activate inflammatory cascades and leucocytes and could play a role in multiple organ failure. Further studies are needed to evaluate the implications of changes in plasma trypsin in the disease process of equine acute abdomen and to demonstrate possible pancreatic damage. PMID- 12090771 TI - Testicular damage from anabolic treatments with the beta(2)-adrenergic agonist clenbuterol in pigs: a light and electron microscope study. AB - The morphological consequences of anabolic clenbuterol treatment on the testicular parenchyma were investigated in 30 pigs at morphological and ultrastructural levels. Clenbuterol was given with food (1 ppm). In the first group (n=10), treatment was maintained until slaughter (experimental period 3 months). In the second group (n=10), clenbuterol was withdrawn 2 weeks before slaughter (experimental period 2.5 months). A third group (n=10) of pigs not fed with clenbuterol served as controls. Animals were slaughtered at 9 months of age and samples of testicular parenchyma were collected for light and electron microscope studies. In the clenbuterol-treated groups, the interstitial cells showed a considerable increase in the organelles involved in testosterone production, with an increased development of the mitochondria, smooth endoplasmic reticulum, Golgi apparatus and lipid droplets compared to the control group. The seminal epithelium displayed many lipid vacuoles and evident signs of tubular involution, such as degenerating and multinucleate germ cells. Sertoli cells gave evidence of metabolic alterations such as large lipid deposits and cytolysosomes. PMID- 12090772 TI - Detection of Brucella species in the milk of infected cattle, sheep, goats and camels by PCR. AB - One hundred and three milk samples were collected from 52 cows, 21 ewes, 18 goats and 12 camels. The animals tested positive to at least one of the following: (1) standard tube agglutination test (SAT); (2) Rose Bengal plate test (RBPT); (3) milk ring test (MRT). All milk samples were examined by culture and single-step polymerase chain reaction (PCR) techniques for detection of Brucella species. The PCR assay amplified Brucella-DNA from 29 bovine milk samples, 10 from sheep, 13 from goats and one from a camel. The direct culture method detected Brucella organisms from 24 samples of cows' milk, 12 from sheep, 10 from goats and failed to detect any Brucella organisms from camels' milk. PCR detected up to 100 colony forming units (CFU) of B. abortus per millilitre of milk in 100% of diluted milk samples, and 1000 CFU of B. melitensis from 70% of milk samples. Although the overall sensitivity of the PCR was higher than the culture method, it should be possible to increase the sensitivity to detect lower numbers of Brucella organisms in field samples. The speed and sensitivity of the PCR assay suggest that this technique could be useful for detection of Brucella organisms in bovine milk, as well as in sheep, goat, and camels milk. PMID- 12090773 TI - The control of Varroa destructor using oxalic acid. AB - Twenty-four honeybee (Apis mellifera) colonies were used to monitor the efficacy of a solution of 2.9% oxalic acid (OA) and 31.9% sugar against the mite Varroa destructor. Mite mortality was established prior to and after OA treatments, which were conducted in August and September. The treatments resulted in 37% mite mortality as opposed to 1.11% in the controls. OA treatment conducted in September on previously untreated colonies resulted in 25% mite mortality. OA treatments in October and November resulted in approximately 97% mite mortality. These results suggest that OA is effective during the broodless period and less effective when applied to colonies with capped broods. The possible use of OA against the Varroa mite in honeybee colonies as an alternative to routine chemical treatments is discussed. PMID- 12090774 TI - Further contributions to the genetic aspect of congenital sensorineural deafness in Dalmatians. AB - Sensorineural deafness is a common congenital disorder in Dalmatians and is genetically transmitted. Different modes of inheritance have been proposed and the objective of this study was to study these by segregation analyses using maximum likelihood procedures. Data from 33 complete Dalmatian families were collected and data from 56 single Dalmatians added. This resulted in a total of 575 dogs with 357 known phenotypes. All dogs were clinically evaluated and electrophysiologically tested with brainstem auditory evoked responses. The prevalence of deafness was 16.5% (9.4% unilaterally deaf, 7.1% bilaterally deaf). Females were 4.4% more affected than males but this difference was not significant. Within the same litter, different phenotypic expressions of deafness occurred, which suggested different expressions of the disease. In addition, two data sets were analysed: the first included normal, uni- and bilaterally deaf dogs, the second had normal and deaf Dalmatians. We found that a recessive allele at a single biallelic major locus fitted our data best, although an incomplete penetrance of the recessive homozygotes was observed. PMID- 12090775 TI - The use of cardboard bedding material as part of an environmental control regime for heaves-affected horses: in vitro assessment of airborne dust and aeroallergen concentration and in vivo effects on lung function. AB - This study aimed to test whether shredded cardboard is an appropriate minimum dust bedding material for heaves-affected horses. Results of standardized in vitro measurement of airborne dust and aeroallergen concentrations of cardboard bedding were significantly lower than those of common bedding materials. Six heaves-affected horses in clinical remission after pasturing were stabled for two months on cardboard bedding and fed grass silage. Pulmonary function tests (PFT: ventilatory mechanics, arterial blood gases, airway inflammation scoring, bronchoalveolar cytology) were performed before, during and after this period and after stabling the horses in poor hygienic conditions. PFT values measured during and after the stabling period on cardboard bedding were not significantly different from those recorded after the period at pasture or from those of healthy horses, but were significantly different from those recorded in poor hygienic conditions. On basis of the in vitro and in vivo results it can be concluded that cardboard bedding, used in conjunction with low-dust forage, may be appropriate in the provision of minimum-dust management of heaves-affected horses. PMID- 12090776 TI - N-ras mutation in a feline lymphoma. Low frequency of N-ras mutations in a series of feline, canine and bovine lymphomas. PMID- 12090777 TI - Investigation of pre-natal Chlamydophila abortus (Chlamydia psittaci) exposure of female lambs and the outcome of their first pregnancy. PMID- 12090778 TI - Lung carcinosarcoma in a dog: gross and microscopic examination. PMID- 12090779 TI - Comparison of epidural anaesthesia with lignocaine or xylazine in cats. PMID- 12090780 TI - Mediators of asthma: nitric oxide. AB - Endogenous nitric oxide is an ubiquitous gaseous molecule that regulates many aspects of human airway biology including the modulation of airway and vascular smooth muscle tone. It is generated from the three different enzymes nitric oxide synthases (NOS) -1, -2 and -3 which are all expressed in pulmonary cells. NOS-1 is localised primarily to neuronal structures, where NO is a mediator of the inhibitory Non-Adrenergic Non-Cholinergic System and NOS-3 is present in endothelial cells. While these enzymes are constitutively expressed, NOS-2 is an inducible enzyme independent of calcium and highly induced in inflammatory diseases such as allergic asthma, where NO may act beneficial or deleterious depending on the site of and amount of generation. The use of NO-donor compounds or classical unselective NOS inhibitors did not lead to significant therapeutical effects in asthmatic patients. Insights on the precise role of NO in asthma can only be achieved by targeting NO generation selectively. More potent and selective NOS-2 inhibitors have to clarify a role of NOS-modification based therapy in clinical routine. NO can also be detected in the exhaled air. Increased levels of exhaled NO in asthmatic patients may be useful for a non invasive determination of airway inflammation. PMID- 12090781 TI - Pharmacology of acute lung injury. AB - The acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) is a clinical syndrome that affects both medical and surgical patients. To date, despite improved understanding of the pathogenesis of ALI/ARDS, pharmacological modalities have been unsuccessful in decreasing mortality. However, several pharmacological agents for ARDS are in development and have shown great promise. In addition to the anti-inflammatory category including late corticosteroids, inhaled nitric oxide, alveolar surfactant, and vasodilators are being evaluated. Replacements of anticoagulation mediators have also suggested beneficial effects on the patient outcome. This article provides an overview of pharmacological treatments of ALI/ARDS. PMID- 12090782 TI - Onset of action following formoterol Turbuhaler and salbutamol pMDI in reversible chronic airway obstruction. AB - Short-acting beta(2)-agonists are currently recommended for symptom relief in asthma and the treatment of mild, acute exacerbations in COPD. However, formoterol has as fast an onset of action as salbutamol with the additional benefit of longer-lasting bronchodilation (approximately 12 h). Furthermore, systemic side effects observed with formoterol are of a similar duration but less pronounced than with short-acting beta(2)-agonists. In this double-blind, randomized, cross-over study, 20 adult patients with reversible chronic airway obstruction (intrinsic asthma or COPD) inhaled single doses of formoterol 9 microg or salbutamol 100 microg (group A) or formoterol 18 microg or salbutamol 200 microg (group B). FEV(1) was measured prior to and 5, 10, 15, 20, 25 and 30 min following inhalation of study drug. No significant differences in FEV(1) values were observed between group A (P=0.704) or group B (P=0.270) at baseline, or at 5 (Group A: P=0.340; Group B: P=0.559) and 15 min (Group A: P=0.526; Group B: P=0.818) post dose. No adverse events were reported during the study. Formoterol Turbuhaler has as rapid an onset of action as salbutamol pMDI when given at the recommended doses. PMID- 12090783 TI - The selective CXCR2 antagonist SB272844 blocks interleukin-8 and growth-related oncogene-alpha-mediated inhibition of spontaneous neutrophil apoptosis. AB - The aims of this study were to investigate the effects of Interleukin-8 (IL-8) and Growth related oncogene-alpha (Gro-alpha) on neutrophil apoptosis and determine the potential for a selective CXCR2 antagonist to inhibit these responses. IL-8 and Gro-alpha both produced dose dependent inhibition of spontaneous human neutrophil apoptosis after 16 hours incubation (59+/-3.5% and 27.5+/-3% respectively; EC50s 2.2+/-1.8 nM, and 0.5+/-0.2 nM respectively). The pro-survival effect of a fixed concentration of agonist (IL-8 or Gro-alpha) on cultured neutrophils was abrogated by a selective CXCR2 antagonist SB272844 (K(D)s 253 nM and 49.9 nM in the presence of IL-8 or Gro-alpha respectively). Our data suggests that the anti-apoptotic effect of Gro-alpha is mediated through CXCR2 as selective CXCR2 blockade with SB272844 can potently abrogate this response. The inhibitory effect of IL-8 may in addition partly be mediated through CXCR1 as SB272844 was less potent in its ability to abrogate the anti apoptotic effects of IL-8 when this agent was used as an agonist. CXCR2 antagonists may have a therapeutic role in controlling neutrophil-driven inflammation by reducing neutrophil recruitment and restoring neutrophils to the tissue clearance pathway of apoptosis. PMID- 12090784 TI - Ozone exposure may enhance airway smooth muscle contraction by increasing Ca(2+) refilling of sarcoplasmic reticulum in guinea pig. AB - Ozone induces airway hyperresponsiveness, but there is controversy about effects of ozone on smooth muscle per se. We therefore investigated effects of in vivo ozone exposure on intracellular calcium mobilization in relation to tracheal smooth muscle contractility in the guinea pig in vitro. Guinea pigs underwent ozone exposure or sham exposure (3 ppm, 2 h). Then, a tracheal smooth muscle strip was mounted in an organ bath to record isometric tension. Effects of ozone exposure on acetylcholine-induced contraction of smooth muscle were as follows. Contraction was not altered in normal Krebs solution, but was increased in Ca(2+) free solution in ozone-exposed animals. Decline of tension on repetitive application of acetylcholine in Ca(2+)-free solution was reduced, while the tension decline rate while acetylcholine was washed out with Ca(2+)-free solution was facilitated in ozone-exposed animals. Tension decline during the continuous administration of acetylcholine in Ca(2+)-free solution was slowed. Contraction occurred more quickly in Ca(2+)-free solution in ozone-exposed animals. Results suggest that ozone has a direct action on airway smooth muscle by changing Ca(2+) mobilization; Ca(2+) refilling via a Ca(2+) pump and Ca(2+) release via Ca(2+) channels in the sarcoplasmic reticulum were increased, while Ca(2+) extrusion via the plasma membrane Ca(2+) pump was unchanged. PMID- 12090785 TI - New evidence for transmitter role of VIP in the airways: impaired relaxation by a catalytic antibody. AB - The identity of the transmitter(s) of nonadrenergic, noncholinergic airway smooth muscle relaxation has long been investigated. Recently, nitric oxide (NO) has been proposed as the main, if not the only transmitter. We earlier suggested vasoactive intestinal peptide (VIP) as a candidate transmitter and target for pathogenic catalytic autoantibodies (VIPases) found in certain humans. To re examine the role of VIP, we studied the airway transport and effects of a model monoclonal antibody (Ab) capable of binding and cleaving VIP. In vitro receptor binding assays indicated the catalytic light chain subunit of the VIPase Ab to inhibit the saturable binding of (Tyr(10-125)I) VIP by guinea pig lung membranes, whereas a catalytically deficient mutant of the Ab light chain was without significant inhibitory activity. Systemically administered IgG preparations of the VIPase Ab accumulated in the airway lavage fluid of guinea pigs at levels close to those in blood, suggesting that the Ab reaches the airways freely. Electrical field stimulation (EFS)-induced relaxations of tracheal strips were weaker and shorter in VIPase-treated animals than in control nonimmune IgG treated animals. The inhibitory effect of the VIPase was dose-dependent. VIPase mediated inhibition of EFS-induced relaxation was evident both in the absence and presence of blockade of beta-adrenergic and cholinergic receptors. Thus, circulating VIP binding and cleaving antibodies can reach the airways and attenuate the neurogenic relaxation of guinea pig tracheal smooth muscle, probably by neutralizing endogenously released VIP. The findings support a role for VIP as a major mediator of neurogenic relaxation of guinea pig tracheal smooth muscle. Lack of complete abrogation of relaxation is consistent with a co transmitter role for NO. PMID- 12090786 TI - Intranasal fluticasone propionate inhibits allergen induced bone marrow eosinophilia in mice. AB - Local corticosteroids are currently the most efficient safe anti-allergic treatment, which attenuate eosinophilic tissue inflammation through several mechanisms. We evaluated the effect of local airways corticosteroid on repeated allergen exposure-induced bone marrow activation and airway eosinophilia using the number of eosinophils in bone marrow, bronchoalveolar lavage fluid (BALf) and airways tissue as study end-points. Male BALB/c mice were sensitized by intraperitoneal injections of aluminum-precipitated ovalbumin (OVA) on two different days (5 days apart). Eight days after the second sensitization, the animals were challenged intranasally with OVA or phosphate-buffered saline (PBS) on 5 consecutive days. Concomitantly with challenges mice were treated with fluticasone propionate or respective vehicle. OVA exposures induced a significant increase in eosinophil numbers in bone marrow, BALf and airways tissue (P<0.005). Treatment with fluticasone propionate significantly reduced the increase of absolute number of mature bone marrow eosinophils (P=0.014) and showed a tendency towards decrease in the immature bone marrow eosinophil number (P=0.057) compared to controls. However, fluticasone propionate had no significant effect on BALf and airways tissue eosinophils (P=0.28 and 0.07, respectively). In this murine allergy model intranasal corticosteroid reduced number of bone marrow mature eosinophils, but did not significantly affect airways cell populations. PMID- 12090787 TI - Biological actions of formoterol isomers. AB - Racemic beta(2) agonists, composed of equal amounts of (R)- and (S)-isomers, can display anomalous actions that compromise their effectiveness as asthma therapies. Loss of efficacy during regular use is characteristic of isoprenaline, albuterol and terbutaline and has in part been attributed to the biological effects of the (S)-isomer. This hypothesis was applied to the (R,R)- and (S,S) isomers of formoterol. (R,R)-formoterol had 1000-times greater affinity (2.9 nm) to the human beta(2) adrenoceptor than (S,S)-formoterol (3100 nm), with receptor binding modulating intracellular cAMP levels. The minimum lethal intravenous (IV) dose was determined to be 100 mg/kg for (R,R)- and 50 mg/kg for (S,S)-formoterol, suggesting that the toxicity of (S,S)-formoterol may not be related to the binding of beta(2) adrenoceptors. In tissues pretreated with (S,S)-formoterol but not with (R,R)- or racemic formoterol contractions to high concentrations of carbachol were exaggerated. In vivo experiments with sensitized guinea pigs demonstrated that (R,R)-formoterol inhibited both histamine and antigen-induced bronchoconstriction with greater potency than (R,R/S,S)-formoterol while (S,S) formoterol was ineffective. Metabolic radiolabeling experiments of (R,R)-, (S,S)- or (R,R/S,S)-formoterol with crude human liver phenolsulfotransferase (PST) determined the V(max)/K(m) values to be (0.151), (0.74) and (0.143), respectively. The reciprocal plot illustrates a 2-fold reduction in sulfation rate when (R,R)-formoterol is present as a single isomer. The data presented here suggest that (R,R)-formoterol binds to the beta(2) adrenoceptor and inhibits the contraction of bronchial tissues by spasmogens. However, (S,S)-formoterol exhibits properties inconsistent as an asthma therapeutic and may antagonize the actions of (R,R)-formoterol. PMID- 12090789 TI - Analysis of the stability of stored adenosine 5'-monophosphate used for bronchoprovocation. AB - Adenosine 5'-monophosphate (AMP) bronchial challenge has been shown to be very useful tool in the diagnosis of asthma. Freshly test solutions are prepared just prior to each test in most of the studies. The objective of this study was to assess the stability of AMP solutions at different temperatures using a reversed phase high-performance liquid chromatography assay. Sodium salt AMP solutions in concentrations of 0.03 mg/ml and 400 mg/ml were analyzed. One aliquot was kept at room temperature (20-25 degrees C) and the others were refrigerated at 4 degrees C. Room temperature stored samples were analyzed daily. Refrigerated stored samples were analyzed daily for first 15 days and then weekly. The duration of the study was 25 weeks. Samples were injected into the chromatograph column in quadruplicate and quantification was based on the arithmetic mean and standard deviation (+/-SD) of four measurements. Room temperature stored samples at concentrations of 0.03 mg/ml showed a mean percent variation greater than 10% at day 9 and greater than 75% at day 14. Samples at concentrations of 400 mg/ml maintained almost the initial concentration during the first 10 days, but decomposition occurred thereafter. In contrast, there was no significant degradation of refrigerated stored samples throughout the study period. We conclude the exposure to room temperature of AMP solutions results in a substantial loss of the initial concentration, but the shelf life of adequately prepared stock AMP solutions stored at 4 degrees C is at least 25 weeks. PMID- 12090790 TI - Comparison of effects of anti-IL-3, IL-5 and GM-CSF treatments on eosinophilopoiesis and airway eosinophilia induced by allergen. AB - Allergic inflammation is dominated by eosinophils. IL-3, IL-5, and GM-CSF are involved in production and activation of eosinophils. IL-5 has been reported to be crucial for the induction of airway eosinophilia. However, the contribution of IL-3 and GM-CSF to allergic airway inflammation remains to be determined. To address this issue, ovalbumin-sensitized Balb/c mice were repeatedly exposed to allergen via airway route. Animals were pretreated intraperitoneally with neutralising anti-IL-3, anti-IL-5 and/or anti-GM-CSF antibodies. Newly produced inflammatory cells were pulse-labelled with the thymidine analogue 5-bromo-2' deoxyuridine (BrdU), which is incorporated into DNA during the cell mitosis. BAL and bone marrow cells were collected 24 h after the last allergen exposure, and differential cell counts and immunocytochemical detection of BrdU-labelled cells were performed. Anti-IL-5 strongly reduced both BAL and bone marrow eosinophilia, as well as the number of BrdU-positive BAL-granulocytes. In contrast, anti-IL-3 and anti-GM-CSF alone had little and no inhibitory effect on these responses, respectively. Even the combined treatment with anti-IL-3 and anti-GM-CSF showed only a non-significant tendency to attenuate these responses. These data suggest that the efficacy of treatments with anti-IL-3 and anti-GM-CSF is much weaker than that with anti-IL-5. IL-5 may be the preferred target to block eosinophilia in allergic diseases. PMID- 12090788 TI - Adenosine-mediated bronchoconstriction and lung inflammation in an allergic mouse model. AB - In this study, we studied the role of adenosine on airway responsiveness and airway inflammation using an allergic mouse model. Mice were sensitized by two i.p. injections of ragweed and three consecutive ragweed aerosol challenges. It was found that inhalation of adenosine causes a dose-related bronchoconstriction in this model. Ragweed sensitized and challenged mice showed increased sensitivity to airway challenge to adenosine compared to control animals. Theophylline, a non-selective adenosine receptor antagonist, blocked adenosine induced bronchoconstriction, but was unable to inhibit bronchoconstrictor response to methacholine. Mice systemically sensitized and airway challenged with allergen showed a marked airway inflammation manifesting increases in eosinophils, lymphocytes and neutrophils, and decrease in macrophages. Twenty four hours after airway challenge with allergen, aerosolization of adenosine further potentiated the allergen-induced airway inflammation. Cells in bronchoalveolar lavage fluid after adenosine aerosolization increased by 3.07 fold as compared to control mice, and by 1.8-fold compared to ragweed sensitized and challenged mice. The increases in eosinophils, lymphocytes, and neutrophils caused by allergen were potentiated after adenosine challenge. Unexpectedly, macrophages significantly decreased after adenosine challenge. Theophylline attenuated adenosine-enhanced airway inflammation, but could not reverse allergen induced airway inflammation. These findings suggested that specific adenosine receptors contribute to airway responsiveness and airway inflammation associated with this model of allergic asthma. PMID- 12090791 TI - Role of cholinergic reflexes on the bronchoconstrictor reactivity to neurokinin a in allergic dogs. AB - Neurokinin A (NKA) potentiates airway cholinergic neurotransmission in several species. In this study, the role of cholinergic reflexes on the bronchoconstrictor response to NKA was evaluated in non-sensitized dogs and in allergic dogs neonatally sensitized to ragweed in which heightened bronchoconstrictor reactivity to NKA has previously been observed. Cardiopulmonary functions, including pulmonary resistance (R(L)) were measured in anesthetized, spontaneously breathing dogs before and after increasing concentrations of aerosolized NKA. The provocative concentrations of NKA increasing R(L) by 25% above the baseline (PC(25)) was measured before and after ( approximately 10 min) aerosolized saline or ipratropium bromide (0.01%). This concentration of ipratropium produced a 250-fold shift in the methacholine dose response curve. In sensitized dogs, NKA bronchoconstrictor reactivity (PC(25)=0.050+/-0.011%) was 2.5 times more potent than that of non-sensitized controls (PC(25)=0.177+/-0.031%). Ipratropium bromide inhibited the bronchoconstrictor response to NKA in both sensitized and non-sensitized dogs and after ipratropium, NKA reactivity was 5.2-fold less in allergic dogs (PC(25)=0.246+/-0.048%) as compared to 3.5 fold less in non-sensitized controls (PC(25)=0.622+/-0.106%). In conclusion, cholinergic reflexes are important components of the bronchoconstrictor response to NKA in dogs particularly in those sensitized neonatally to ragweed. It is speculated that heightened activity of cholinergic reflexes contributes to the bronchial hyperresponsiveness seen in allergic dogs. PMID- 12090792 TI - Formoterol Turbuhaler 4.5 microg (delivered dose) has a rapid onset and 12-h duration of bronchodilation. AB - Clinical trials show that formoterol (Oxis) Turbuhaler 4.5 microg delivered dose (6 microg metered dose) has a rapid onset of bronchodilation similar to that of salbutamol and a 12-h duration of action. Maximum increase in FEV(1) and duration of bronchodilation are dose-dependent, the 4.5 microg dose being the lowest dose tested giving both effects. Clinical studies investigating onset of bronchodilation show a significant increase in specific airway conductance occurring within 1 min after inhalation of formoterol Turbuhaler 4.5 microg. When measured from 3-20 min after inhalation, formoterol Turbuhaler 4.5 microg showed similar increases in FEV(1) to salbutamol administered via pMDI. No difference in onset of bronchodilation was observed between the formoterol Turbuhaler 4.5 and 9 microg doses.Single-dose studies and studies of 1-12 weeks' duration show that formoterol Turbuhaler 4.5 microg produces a significant and clinically important mean bronchodilating effect for > or =12 h after inhalation. In the cited studies no significant differences in duration of bronchodilation were observed between the formoterol Turbuhaler 4.5 and 9 microg doses. CONCLUSION: clinical data show that formoterol Turbuhaler 4.5 microg is an effective dose in patients with asthma, with a rapid onset of bronchodilation and a duration of at least 12 h. PMID- 12090793 TI - Environmental controls in operating theatres. AB - Surgical-site infection is the leading complication of surgery. Normal skin flora of patients or healthcare workers causes more than half all infections following clean surgery, but the importance of airborne bacteria in this setting remains controversial. Modern operating theatres have conventional plenum ventilation with filtered air where particles >/=5 microm are removed. For orthopaedic and other implant surgery, laminar-flow systems are used with high-efficiency particulate air (HEPA) filters where particles >/=0.3 microm are removed. The use of ultra-clean air has been shown to reduce infection rates significantly in orthopaedic implant surgery. Few countries have set bacterial threshold limits for conventionally ventilated operating rooms, although most recommend 20 air changes per hour to obtain 50-150 colony forming units/m(3) of air. There are no standardized methods for bacterial air sampling or its frequency. With the use of HEPA filters in operating theatre ventilation, there is a tendency to apply cleanroom technology standards used in industry for hospitals. These are based on measuring the presence of particles of varying sizes and numbers, and are better suited than bacterial sampling. Environmental bacterial sampling in operating theatres should be limited to investigation of epidemics, validation of protocols, or changes made in materials which could influence the microbial content. PMID- 12090794 TI - The control of hospital infection in Portugal. PMID- 12090795 TI - Outbreak of a susceptible strain of Acinetobacter species 13 (sensu Tjernberg and Ursing) in an adult neurosurgical intensive care unit. AB - Between December 1999 and June 2000, an outbreak caused by Acinetobacter emerged on the neurosurgical intensive care unit of our hospital. It was shown using automated ribotyping using Eco RI and pulsed-field gel electrophoresis that the outbreak was caused by spread of a single strain, which was identified by ribotyping and amplified ribosomal DNA restriction analysis as Acinetobacter DNA group 13TU (sensu Tjernberg and Ursing). The outbreak strain, which showed no antibiotic resistance, was identified in 23 patients, five of whom developed an infection. The organism was also isolated from various environmental sites. Cross transmission among patients continued despite contact isolation of colonized patients and reinforcement of basic disinfection procedures. Eventually, after implementation of additional stringent measures such as cohorting of positive patients and daily disinfection of the floor, the outbreak was brought under control. This study demonstrates that apart from Acinetobacter baumanii, Acinetobacter 13TU strains, even when they are fully susceptible, may cause outbreaks that are difficult to control. Correct identification to the species level of Acinetobacter by genotypic methods is necessary to get insight in the importance of the different Acinetobacter genomic species in hospital epidemiology. PMID- 12090796 TI - A rapid and easy PCR-RFLP method for genotyping Serratia marcescens strains isolated in different hospital outbreaks and patient environments in the Lyon area, France. AB - A new genotyping method for Serratia marcescens is described. This method uses the flagellin gene as target for polymerase chain reaction amplification and Alu I restriction fragment length polymorphism. The strains tested belonged to 13 different hospital clusters of S. marcescens isolated between 1983 and 1988, concerning outbreaks and/or patient environments in different hospital units in Lyon and the Rhone-Alpes region of France. Initially, the classification had been performed by marcescinotyping. These strains were then tested by ribotyping and genotyping of the flagellin gene. Genotyping showed similar classification to ribotyping. The genotyping method is the easiest technique, as reproducible as ribotyping, and with almost the same ability to discriminate different strains. It does not need expensive equipment, is more rapid, and is less labor intensive than ribotyping. With this method, all strains of S. marcescens including sporadic isolates could be amplified and typed. Antibiotic sensitivity determination was found to be a useful complementary and confirmation test for all these typing methods. PMID- 12090797 TI - Antibiotic resistance as an indicator of bacterial chlorhexidine susceptibility. AB - The antibiotic and chlorhexidine (CHX) susceptibility of 70 distinct clinical isolates: Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Staphylococcus aureus (not MRSA), Streptococcus pyogenes and Enterococcus faecalis (10 of each) were tested using minimal bactericidal (MBC) and/or minimal inhibitory (MIC) concentrations. Non-fermentative bacteria tolerated CHX at high concentrations; Gram-positive cocci, especially S. pyogenes, were the most susceptible. We found a good correlation between CHX and antibiotic susceptibility in both MIC and MBC among Gram-negative bacteria, and mainly in MBC among Gram-positive bacteria. Resistance to ciprofloxacin, imipenem, cefotaxime, ceftazidime, gentamicin and aztreonam appeared to indicate increased CHX resistance among Gram-negative bacteria. This finding gives clinicians the ability to predict CHX susceptibility according to routine antibiotic resistance testing. PMID- 12090798 TI - Clinical double-blind trial on the dermal tolerance and user acceptability of six alcohol-based hand disinfectants for hygienic hand disinfection. AB - Six commercially available alcohol-based hand rubs [AHD 2000, Desderman, Mucasept A, Manorapid (Poly-Alkohol, Spitacid, and Sterillium] were investigated in a clinical double-blind trial involving 10 participants who had no previous experience of using hand rubs (Group 1), and seven who had substantial professional experience of using hand rubs (Group 2; virology laboratory staff). Group 1 was studied for one week with 20 applications on day 1 and then five applications per day for six days. Transepidermal water loss, dermal water content and superficial sebum content of the skin were measured before and after the seven-day application of the products, as well as user acceptability (self assessment of smell, speed of drying, emolliant effect, skin dryness). Group 2 used each preparation twice for two weeks in a random sequence, and carried out self assessment at the end of each fortnight. Transepidermal water loss (mean baseline: 18.7 g/m(2)h), dermal water content (mean baseline dorsum: 75.6) and superficial sebum content (mean baseline dorsum: 4.8 microg/cm(2)) did not change significantly. In both groups assessments of the smell and the speed of drying did not reveal any significant differences between the six products. Sterillium had the best emollient effect of all products (P<0.05; Wilcoxon test and Mann Whitney-U test) and was significantly better than Desderman, AHD 2000, and Mucasept A, causing less skin dryness after seven days use in Group 1 (P<0.05; Mann-Whitney-U test). Manorapid caused significantly less dryness than Spitacid, AHD 2000, and Mucasept A in Group 2 after the first use, but no significant difference was observed after the second use. Thus alcohol-based hand rubs that contain emolliants, irrespective of the type of alcohol (n-propanol, iso-propanol or ethanol), are well tolerated and do not dry out or irritate the skin. Personal assessments showed significant differences for the emolliant effect and the extent of dryness. Both factors are very important, as user acceptability has an impact on compliance. Sterillium is the only hand disinfectant containing mecetronium etilsulphate which has been shown to have an emolliant effect. Future research should focus on user acceptability in order to improve compliance. PMID- 12090799 TI - The virucidal spectrum of a high concentration alcohol mixture. AB - The virucidal spectrum of a high concentration alcohol mixture (80% ethanol and 5% isopropanol) was determined for a broad series of lipid-enveloped (LE) and non lipid-enveloped (NLE) viruses covering all relevant blood-borne viruses. LE viruses were represented by human immunodeficiency virus (HIV), bovine viral diarrhoea virus (BVDV), a specific model virus for hepatitis C virus (HCV), pseudorabies virus (PRV), and vaccinia virus. For the NLE viruses hepatitis A virus, canine parvovirus (a model for human parvovirus B19), and reovirus type 3 (Reo-3) were used. PRV, vaccinia, and Reo-3 served as general model viruses. The alcohol mixture was spiked with 5% (v/v) virus, mixed and tested for residual virus after 5 min treatment. Complete clearance (reduction by a factor of >10(6)) was observed for LE viruses, whereas incomplete to insignificant clearance (ranging from no reduction up to a maximum factor of 10(4)) was found for NLE viruses. In a second series of spiking experiments using the LE viruses BVDV, HIV, and PRV, complete clearance (reduction by a factor of >10(6)) was found after 20 s treatment. These data strongly suggest that treatment with a high concentration alcohol mixture has a high virucidal potential in particular for the blood-borne LE-viruses HIV, hepatitis B virus, and HCV. Such mixtures are well suited for rapid and frequent disinfection in dental practice being non hazardous and non-toxic. PMID- 12090800 TI - Cost-effectiveness of active surveillance cultures and contact/droplet precautions for control of methicillin-resistant Staphylococcus aureus. AB - Some have reported that adopting Centers for Disease Control and Prevention guidelines requiring contact precautions for methicillin-resistant Staphylococcus aureus (MRSA) had no impact on rates of nosocomial spread or infection, and may therefore waste money. The objective of the present study was to evaluate the cost-effectiveness of active surveillance cultures and barrier precautions for controlling MRSA. Estimated costs of surveillance cultures and isolation measures used during an MRSA outbreak at this hospital were compared with the estimated attributable excess costs of methicillin resistance (i.e., the difference between MRSA and methicillin-sensitive S. aureus costs) for bacteraemias occurring during an MRSA outbreak not promptly controlled at another hospital. The study was set in the neonatal intensive care units of two tertiary care hospitals. Estimated costs of controlling the 10.5-month outbreak in this neonatal intensive care unit that resulted in 18 colonized and four infected infants ranged from $48 617 to $68 637. The estimated attributable excess cost of 75 MRSA bacteraemias in a second neonatal intensive care unit outbreak that resulted in 14 deaths and lasted 51 months was $1 306 600. Weekly active surveillance cultures and isolation of patients with MRSA halted an outbreak at this hospital, and cost 19- to 27-fold less than the attributable costs of MRSA bacteraemias in another outbreak that was not promptly controlled. The costs of infections at other body sites and the human cost of deaths from infection were not estimated but would further help to justify the cost of identifying colonized patients and implementing effective preventive measures. PMID- 12090801 TI - Susceptibility of healthcare workers to measles, mumps rubella and varicella. AB - Susceptible healthcare workers are at risk of acquiring and transmitting measles, mumps, rubella and varicella to or from patients. We tested 333 employees at Padova University Hospital, for specific IgG. Of the workers tested, 98.2% were positive for measles, 85.9% for mumps, 97.6% for rubella and 97.9% for varicella. Eventhough participants born before 1957 were not all positive, they had a higher prevalence of immunity. We suggest serological screening in high-risk areas (paediatrics, oncology, radiotherapy, infectious diseases, maternity and the laboratory) and vaccination of susceptible employees. PMID- 12090802 TI - Hospital-acquired listeriosis. AB - We report four cases of listeriosis that occurred over a two-month period in north east England. Due to the apparent nosocomial acquisition of infection and the clustering of cases in time and place, extended epidemiological investigation was performed and the outbreak was traced to a caterer who was providing sandwiches for hospital shops. We discuss the difficulties in preventing food borne listeriosis in the hospital setting. PMID- 12090803 TI - Contamination of room door handles by methicillin-sensitive/methicillin-resistant Staphylococcus aureus. AB - We investigated the contamination of room door handles by Staphylococcus aureus in wards of a university hospital. Door handles in 53 (27.0%) of 196 rooms were contaminated by methicillin-sensitive Staphylococcus aureus (MSSA) and/or methicillin-resistant Staphylococcus aureus (MRSA); MSSA was detected on door handles of 41 rooms (20.9%), MRSA on door handles of 17 rooms (8.7%), and MSSA and MRSA on the same door handles of five rooms (2.6%). The density of MSSA contamination was 1-2.6x10(4) colony forming units (cfu)/door handle, and that of MRSA was 1-6.0x10(3) cfu/door handle. The MRSA contamination rate on door handles of rooms with patients with MRSA was 19.0% (4/21 rooms) while that on door handles of rooms with patients without MRSA was 7.4% (13/175); the difference was not significant. These results suggest extensive contamination of MSSA and MRSA in the hospital environment. PMID- 12090804 TI - Comparing microbial performance of impactor air samplers: an uneasy task. PMID- 12090805 TI - Biocides and antibiotics with apparently similar actions on bacteria: is there the potential for cross-resistance? PMID- 12090806 TI - Postoperative endophthalmitis: the application of hazard analysis critical control points to an infection control problem. PMID- 12090807 TI - Is brushing of central venous catheters safe clinical practice? PMID- 12090808 TI - Quality of final rinse water used in washer-disinfectors for endoscopes. PMID- 12090809 TI - Infection control hazards of intraoperative forced air warming. PMID- 12090810 TI - Failure to heal should replace infection rate when monitoring surgical wounds. PMID- 12090811 TI - Do lithium and anticonvulsants target the brain arachidonic acid cascade in bipolar disorder? AB - BACKGROUND: Lithium and certain anticonvulsants, including carbamazepine and valproic acid, are effective antimanic drugs for treating bipolar disorder, but their mechanisms of action remain uncertain. EXPERIMENTAL OBSERVATIONS: Feeding rats lithium chloride for 6 weeks, to produce a brain lithium concentration of 0.7mM, reduced arachidonic acid turnover within brain phospholipids by 75%. The effect was highly specific, as turnover rates of docosahexaenoic acid and palmitic acid were unaffected. Arachidonate turnover in rat brain also was reduced by long-term valproic acid administration. Lithium's reduction of arachidonate turnover corresponded to its down-regulating gene expression and enzyme activity of cytosolic phospholipase A(2), an enzyme that selectively liberates arachidonic but not docosahexaenoic acid from phospholipids. Lithium also reduced the brain protein level and activity of cyclooxygenase 2, as well as the brain concentration of prostaglandin E(2), an arachidonate metabolite produced via cyclooxygenase 2. CONCLUSIONS: These results give rise to the hypothesis that lithium and antimanic anticonvulsants act by targeting parts of the "arachidonic acid cascade," which may be functionally hyperactive in mania. Thus, drugs that target enzymes in the cascade, such as cyclooxygenase 2 inhibitors, might be candidate treatments for mania. Also, in view of competition between arachidonic and docosahexaenoic acids in a number of functional processes, docosahexaenoic acid or its precursors would be expected to be therapeutic. Neither of these predictions is evident from other current hypotheses for the antimanic action of lithium and anticonvulsant drugs. PMID- 12090812 TI - The neural basis of mood-congruent processing biases in depression. AB - BACKGROUND: Mood-congruent processing biases are among the most robust research findings in neuropsychological studies of depression. Depressed patients show preferential processing of negatively toned stimuli across a range of cognitive tasks. The present study aimed to determine whether these behavioral abnormalities are associated with specific neural substrates. METHODS: Ten depressed patients and 11 healthy control subjects underwent scanning during performance of an emotional go/no-go task using functional magnetic resonance imaging. The task allowed comparison among neural response to happy, sad, and neutral words, in the context of these words as targets (ie, stimuli to which subjects were required to make a motor response) or distractors (ie, stimuli to which the motor response was withheld). RESULTS: Depressed patients showed attenuated neural responses to emotional relative to neutral targets in ventral cingulate and posterior orbitofrontal cortices. However, patients showed elevated responses specific to sad targets in rostral anterior cingulate extending to anterior medial prefrontal cortex. Unlike controls, patients showed differential neural response to emotional, particularly sad, distractors in the lateral orbitofrontal cortex. CONCLUSIONS: These findings suggest a distinct neural substrate for mood-congruent processing biases in performance. The medial and orbital prefrontal regions may play a key role in mediating the interaction between mood and cognition in affective disorder. PMID- 12090813 TI - The natural history of late-life depression: a 6-year prospective study in the community. AB - BACKGROUND: Accurate assessment of the natural history of late-life depression requires frequent observation over time. In later life, depressive disorders fulfilling rigorous diagnostic criteria are relatively rare, while subthreshold disorders are common. The primary aim was to study the natural history of late life depression, systematically comparing those who did with those who did not fulfill rigorous diagnostic criteria. METHODS: Within the Longitudinal Aging Study Amsterdam, a large cohort of depressed elderly persons (n = 277) was identified and followed up for 6 years, using 14 observations. Depression was measured using self-reports (the Center for Epidemiological Studies Depression Scale) and diagnostic interviews (the Diagnostic Interview Schedule). The natural history was assessed for symptom severity (Center for Epidemiological Studies Depression Scale score), symptom duration, clinical course type, and stability of diagnoses. RESULTS: The average symptom severity remained above the 85th percentile of the population average for 6 years. Symptoms were short-lived in only 14%. There were remissions in 23%, an unfavorable but fluctuating course in 44%, and a severe chronic course in 32% (percentages do not total 100 because of rounding). Comparing the outcome, there was a clear gradient in which those with subthreshold disorders had the best outcome, followed by those with major depressive disorder, dysthymic disorder, and double depression. However, the prognosis of subthreshold disorders was unfavorable in most cases, while this group was at high risk of developing DSM affective disorders. CONCLUSIONS: The natural history of late-life depression in the community is poor. DSM affective disorders are relatively rare among elderly persons, but do identify those with the worst prognosis. However, subthreshold depression is serious and chronic in many cases. PMID- 12090814 TI - Association between serotonin transporter gene promoter polymorphism (5HTTLPR) and behavioral responses to tryptophan depletion in healthy women with and without family history of depression. AB - BACKGROUND: Evidence suggests that serotonin transporter gene promoter polymorphism (5HTTLPR)-dependent low transcriptional activity of the human serotonin transporter gene may be a genetic susceptibility factor for depression. We studied the behavioral responses to tryptophan depletion (TD) in healthy women with and without a first-degree family history of depression and examined the relationship to 5HTTLPR alleles. METHODS: Twenty-four healthy women with a negative family history of depression and 21 women with a positive family history of depression were genotyped for the polymorphism of the 5HTTLPR and then entered a double-blind, placebo-controlled, randomized crossover TD study. The effects of these interventions were assessed with measures of depression and plasma tryptophan levels. RESULTS: The TD induced a robust decrease of plasma tryptophan levels in all women irrespective of family history of depression or 5HTTLPR genotypes. The s/s genotype of the 5HTTLPR was associated with an increased risk of developing depressive symptoms during TD irrespective of family history. In contrast, individuals with the l/l genotype did not develop depressive symptoms, irrespective of family history. Finally, s/l subjects without family history showed a mood response that was intermediate between the s/s and l/l subjects, while s/l subjects with a family history of depression showed the same depressiogenic effect of TD as seen in the s/s subjects. CONCLUSIONS: The results of the present study suggest that the s-allele of the 5HTTLPR and a positive family history of depression are additive risk factors for the development of depression during TD. PMID- 12090815 TI - Blunted prefrontal cortical 18fluorodeoxyglucose positron emission tomography response to meta-chlorophenylpiperazine in impulsive aggression. AB - BACKGROUND: Impulsive aggression is a prevalent problem and yet little is known about its neurobiology. Preclinical and human studies suggest that the orbital frontal cortex and anterior cingulate cortex play an inhibitory role in the regulation of aggression. METHODS: Using positron emission tomography, regional metabolic activity in response to a serotonergic stimulus, meta chlorophenylpiperazine (m-CPP), was examined in 13 subjects with impulsive aggression and 13 normal controls. The anterior cingulate and medial orbitofrontal regions were hypothesized to respond differentially to m-CPP in patients and controls. In the frontal cortex, regional metabolic glucose response to m-CPP was entered into a group (impulsive aggressive, control) x slice (dorsal, middle, orbital) x position (medial, lateral) x location (anterior, posterior) x hemisphere (right, left) mixed-factorial analysis of variance design. A separate group (impulsive aggressive, controls) x anteroposterior location (Brodmann areas 25, 24, 31, 29) x hemisphere (right, left) analysis of variance was used to examine regional glucose metabolism in the cingulate gyrus. RESULTS: Unlike normal subjects, patients with impulsive aggression did not show activation specifically in the left anteromedial orbital cortex in response to m CPP. The anterior cingulate, normally activated by m-CPP, was deactivated in patients; in contrast, the posterior cingulate gyrus was activated in patients and deactivated in controls. CONCLUSIONS: The decreased activation of inhibitory regions in patients with impulsive aggression in response to a serotonergic stimulus may contribute to their difficulty in modulating aggressive impulses. PMID- 12090817 TI - N400 and automatic semantic processing abnormalities in patients with schizophrenia. AB - BACKGROUND: One factor hypothesized to underlie thinking disturbance in patients with schizophrenia is abnormal or disinhibited automatic spreading activation of semantic networks, which can be assessed using the N400 event-related potential. N400 is a negative-going component elicited at about 400 milliseconds following semantic stimuli that are not primed by the preceding context. Semantic priming refers to facilitated semantic processing gained through preexposure to semantic context, which can happen automatically or strategically. Using N400, inferences can be drawn regarding the extent to which a given context primes a word. METHODS: During a picture-word matching task, N400s to primed (exact match) and unprimed (remotely related) words were recorded from 18 healthy control subjects and 18 patients with schizophrenia performing a picture-word matching task. A short interval (325 milliseconds) between picture and word onset (stimulus-onset asynchrony) was used to optimize the role of automatic spreading semantic activation and to minimize the role of attention, expectancy, preparation, and working memory. RESULTS: Despite behavioral evidence of normal semantic priming, patients generated an abnormally small N400 (ie, less negative) to unprimed words. The N400 to primed words was neither larger nor smaller in patients than in controls, suggesting normal use of context. CONCLUSIONS: A reduced N400 to unprimed words in patients with schizophrenia suggests that there was inappropriate priming of words by remotely related semantic contexts. This is consistent with an overly broad automatic spread of activation through semantic networks in patients with schizophrenia. PMID- 12090816 TI - Gene expression profile for schizophrenia: discrete neuron transcription patterns in the entorhinal cortex. AB - BACKGROUND: Several lines of evidence indicate the altered function of the temporal lobe, including the hippocampus and entorhinal cortex (EC), is associated with schizophrenia. We used single-cell gene expression technologies to assess coordinate changes in the expression of multiple genes, including neuronal signaling and synaptic-related markers in EC layer II stellate neurons. METHODS: We used a single-neuron microdissection technique coupled with linear antisense RNA amplification and high density/candidate gene arrays to assess coordinate changes in gene expression. The expression and relative abundance of more than 18,000 messenger RNAs were assessed from EC layer II stellate neurons from postmortem samples of schizophrenic and age-matched control brains. Results of this initial screen were used to perform a more specific secondary messenger RNA screen for each subject. RESULTS: Data disclosed marked differences in expression of various G-protein-coupled receptor-signaling transcripts, glutamate receptor subunits, synaptic proteins, and other transcripts. Results of secondary screening showed significant decreases in levels of G-protein subunit i(alpha)1, glutamate receptor 3, N-methyl-D-aspartate receptor 1, synaptophysin, and sensory nerve action potentials 23 and 25 in the stellate neurons of schizophrenic patients. We observed down-regulation of phospholemman (a phosphoprotein associated with anion channel formation) messenger RNA and protein levels in layer II/III stellate neurons in the population with schizophrenia. CONCLUSIONS: These results provide a preliminary expression profile of schizophrenia in defined neuronal populations. Understanding the coordinated involvement of multiple genes in human disease provides insight into the molecular basis of the disease and offers new targets for pharmacotherapeutic intervention. PMID- 12090819 TI - Recruiting and retaining future generations of physician scientists in mental health. AB - The authors discuss 6 challenges facing the recruitment and retention of physician scientists as career mental health researchers. These challenges include (1) early identification and recruitment at the undergraduate and medical student level; (2) recruitment of a more diverse group of trainees; (3) safety nets for reducing attrition; (4) strategies to promote successful competition for K awards; (5) definition of appropriate roles and career development opportunities in multisite clinical trials; and (6) strategies for the mentoring "cost." A coalition of stakeholders--federal, academic, foundational, and in the pharmaceutical industry--is needed to meet these challenges. PMID- 12090818 TI - A double-blind comparison of desipramine and placebo in children and adolescents with chronic tic disorder and comorbid attention-deficit/hyperactivity disorder. AB - BACKGROUND: Currently, there is no consensus on the best therapeutic approach to chronic tic disorders and comorbid attention-deficit/hyperactivity disorder (ADHD). To address this issue, we evaluated the tolerability and efficacy of the noradrenergic tricyclic antidepressant desipramine hydrochloride in the treatment of children and adolescents with chronic tic disorders and comorbid ADHD. METHODS: Forty-one children and adolescents with chronic tic disorders, including Tourette disorder and comorbid ADHD, were studied in a 6-week, double-blind, placebo-controlled, parallel trial. Desipramine was titrated weekly up to 3.5 mg/kg per day. We rated ADHD and tic symptoms weekly and monitored adverse effects, laboratory findings, and cardiovascular parameters. RESULTS: Treatment with desipramine (mean total daily dose, 3.4 mg/kg per day) was well tolerated without meaningful adverse effects. Desipramine significantly reduced core symptoms of ADHD (ADHD Rating Scale; 42% decrease from baseline relative to placebo, P<.001), with equal response in inattentive symptoms and hyperactive/impulsive symptoms (P<.001 for both). The ADHD response rate was robust (71% vs 0%; desipramine vs placebo, P<.001). Likewise, desipramine significantly reduced tic symptoms (Yale Global Tic Severity Scale; 30% decrease from baseline relative to placebo, P<.001), with equal response in motor and phonic tic symptoms (P<.01 for both). The tic response rate was substantial (58% vs 5%; desipramine vs placebo, P<.001). There were small but statistically significant differences between desipramine and placebo in heart rate and blood pressure. CONCLUSIONS: Treatment with desipramine was well tolerated and was associated with robust clinically significant reductions in tic and ADHD symptoms in children and adolescents with chronic tic disorders and ADHD diagnoses. PMID- 12090820 TI - Mirtazapine in the treatment of panic disorder. PMID- 12090821 TI - Catechol-O-methyltransferase Val-108/158-Met gene variants associated with performance on the Wisconsin Card Sorting Test. PMID- 12090822 TI - Effects of NMDA receptor antagonists: implications for the pathophysiology of schizophrenia. PMID- 12090823 TI - Can schizophrenia be diagnosed in the initial prodromal phase? PMID- 12090824 TI - Substance use and the development of antisocial personality in depressed adolescents. PMID- 12090826 TI - Dolor aqui? Fiebre?: a little knowledge requires caution. PMID- 12090827 TI - Research and human subjects. PMID- 12090828 TI - Grief-love: contradictions in the lives of fathers of children with disabilities. PMID- 12090829 TI - Very early exposure to erythromycin and infantile hypertrophic pyloric stenosis. AB - OBJECTIVE: To assess the link between very early erythromycin exposure and pyloric stenosis in young infants. DESIGN: Retrospective cohort study. PARTICIPANTS AND METHODS: Medicaid or TennCare (Tennessee's program for Medicaid enrollees and uninsured individuals) births in Tennessee from 1985 to 1997. Cases of infants with a hospital discharge diagnosis of pyloric stenosis and an associated surgical procedure code were used. Erythromycin exposure and other antibiotic exposure between 3 and 90 days of life were identified from prescription files. MAIN OUTCOME MEASURES: Hospital discharge diagnosis of pyloric stenosis, and an associated surgical procedure code. RESULTS: Of 933 239 births in Tennessee during the study period, 314 029 were enrolled in Medicaid. Among these infants, 804 (2.6/1000 infants) met the criteria for pyloric stenosis. Very early exposure to erythromycin (between 3 and 13 days of life) was associated with a nearly 8-fold increased risk of pyloric stenosis (adjusted incident rate ratio, 7.88; 95% confidence interval, 1.97-31.57). No increased risk of pyloric stenosis was seen in infants exposed to erythromycin after 13 days of life or in infants exposed to antibiotics other than erythromycin. CONCLUSIONS: The significant increase in pyloric stenosis in children with very early exposure to erythromycin is consistent with reports of other investigators. The risks and benefits of erythromycin should be weighed carefully prior to initiating such therapy in young infants. PMID- 12090830 TI - Type 2 diabetes mellitus in children: prenatal and early infancy risk factors among native canadians. AB - BACKGROUND: Type 2 diabetes mellitus is increasingly being observed among children and youth, including the Native population of Canada. Only one study has investigated prenatal and early infancy risk factors for the disease. METHODS: A case-control study was conducted; 46 patients younger than 18 years were recruited from the only clinical center for the treatment of diabetes serving the province of Manitoba, and 92 age- and sex-matched controls were recruited from a pediatric ambulatory clinic serving a large Native population in Winnipeg, Manitoba. Information on exposure to prenatal and early infancy risk factors was obtained through questionnaires administered by a Native nurse-interviewer. RESULTS: Multiple logistic regression modeling identified preexisting diabetes (odds ratio [OR], 14.4; 95% confidence interval [CI], 2.86-72.5), gestational diabetes (OR, 4.40; 95% CI, 1.38-14.1), and breastfeeding longer than 12 months (OR, 0.24; 95% CI, 0.13-0.99) as significant independent predictors of diabetic status. Other factors, such as low (<2500 g) and high (>4000 g) birth weight and maternal obesity, were also associated with diabetes in our population, but the elevated risks were not statistically significant. CONCLUSION: Breastfeeding reduces the risk of type 2 diabetes among Native Canadian children and should be promoted as a potential intervention to control the disease. PMID- 12090831 TI - Helmet use: a survey of 4 common childhood leisure activities. AB - OBJECTIVE: To compare the distribution and observation of helmet use patterns among 4 common childhood leisure activities-bicycle riding, in-line skating, skateboarding, and scooter riding-in a setting with no helmet use legislation. DESIGN: A cross-sectional, unobtrusive, direct observation of children engaged in 1 of the 4 activities was conducted during an 8-week period. SETTING: Eight communities of at least 1000 persons each in a 2-county Central Texas area. MAIN OUTCOME MEASURES: Prevalence of activity, helmet use, and correct helmet use. RESULTS: Of 841 children observed, most rode bicycles (74.8%); only 4.2% rode skateboards. They included 127 children estimated to be younger than 6 years (15.1%) and 495 estimated to be aged between 6 and 12 years (58.9%). Most were male (67.3%) and observed in urban communities (61.7%). The locations of highest concentration of the 4 activities were minor streets for bicycle riding, playgrounds for in-line skating, and sidewalks for skateboarding and scooter riding. The rate of overall helmet use was 13.6% and did not vary significantly by activity: in-line skating, 18.2%; skateboarding, 14.3%; bicycle riding, 13.5%; and scooter riding, 11.5%. However, the observed correct helmet use prevalence of 72.6% differed significantly by activity. Most children riding scooters wore their helmets incorrectly. Helmet use was positively associated with helmet use by accompanying children and/or adults. CONCLUSIONS: While bicycle riding was the most popular activity observed, helmet use was most prevalent among in-line skaters and least prevalent among scooter riders, many of whom also wore them incorrectly. Observed helmet use was influenced mostly by adult and peer helmet use, indicating that public education and interventions should target children and their parents as well as scooter riders. PMID- 12090833 TI - Rebound in serum bilirubin level following intensive phototherapy. AB - OBJECTIVES: To document the need for repeated phototherapy (as an index of significant rebound in serum bilirubin levels) following the discontinuation of intensive phototherapy and to compare the use of repeated phototherapy in infants who first received phototherapy during their birth hospitalization with the use of first-time phototherapy on readmission after infants were discharged from their birth hospitalization. DESIGN: A retrospective review of the medical records of 303 term and near-term newborns treated between January 1996 and December 1998, who received phototherapy in our well-baby nursery during their birth hospitalization (group 1, n = 158) or who had been discharged from the nursery and were readmitted for phototherapy (group 2, n = 144). All infants received intensive phototherapy but were managed by individual attending pediatricians. Rebound measurements were included if a bilirubin level was obtained between 4 and 48 hours after discontinuing phototherapy. SETTING: Newborn nursery and pediatric ward of a large community hospital. MAIN OUTCOME MEASURES: The number of infants who received repeated phototherapy and the magnitude of the bilirubin-level rebound. RESULTS: Thirteen (8.2%) of 158 (95% confidence interval [CI], 3.9-12.4) infants treated with phototherapy before discharge from the nursery (group 1) and only 1 (0.7%) of 144 (95% CI, 0-2.0) infants who first received phototherapy on readmission (group 2) received repeated phototherapy (P =.002). Phototherapy was discontinued when mean +/- SD total serum bilirubin levels were, 10.4 +/- 1.8 mg/dL (178 +/- 31 micromol/L) in group 1 and 12.3 +/- 1.3 mg/dL (210 +/- 22 micromol/L) in group 2. The mean +/- SD increase in the total serum bilirubin levels following rebound was 1.3 +/- 2.0 mg/dL (22 +/- 34 micromol/L) in group 1 and 0.27 +/- 1.46 mg/dL (4.6 +/- 25 micromol/L) in group 2 (P<.001). CONCLUSIONS: It is not necessary to keep infants in the hospital to check for rebound. However, for infants who require phototherapy during their birth hospitalization and for those with significant hemolytic disease, we recommend obtaining a follow-up bilirubin level 24 hours after discharge. This is probably not necessary in those who are readmitted for phototherapy but, because rare instances of significant rebound have occurred in these infants, additional clinical follow-up is appropriate, particularly if phototherapy is discontinued at higher total serum bilirubin levels than used in this study. PMID- 12090832 TI - The challenge of preventing and treating obesity in low-income, preschool children: perceptions of WIC health care professionals. AB - BACKGROUND: Obesity has become a common nutritional concern among low-income, preschool children, a primary target population of the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC). Nutrition counseling efforts in WIC target childhood obesity, but new approaches are needed that address the different perceptions about obesity that are held by clients and health care professionals. OBJECTIVE: To develop these new approaches, we examined WIC health care professionals' perceptions about the challenges that exist in preventing and managing childhood obesity. DESIGN: A qualitative study using data transcribed from audiotapes of focus groups and individual interviews. We independently read each transcript and coded themes; then, the common themes were selected through group meetings of the authors. SETTING: Kentucky WIC. PARTICIPANTS: Of the 19 health care professionals participating, all had provided nutrition counseling in WIC and all but one were white women. RESULTS: Twelve major themes clustered into 3 domains. The first domain centered on how WIC health care professionals perceived the life experiences, attitudes, and behaviors of the mothers they counseled. They perceived that mothers (1) were focused on surviving their daily, life stresses; (2) used food to cope with these stresses and as a tool in parenting; (3) had difficulty setting limits with their children around food; (4) lacked knowledge about normal child development and eating behavior; (5) were not committed to sustained behavioral change; and (6) did not believe their overweight children were overweight. The second domain described WIC health care professionals' perceptions of counseling interactions. They felt that (7) they might offend mothers when talking about weight, (8) counseling was driven by protocols, and (9) their nutritional advice often conflicted with the advice from the mothers' relatives, friends, or primary care physicians. The last domain described programmatic suggestions WIC health care professionals offered to address childhood obesity: These included (10) promoting a more client-centered approach to counseling, (11) establishing behavioral change goals that were small and endorsed by the mother, and (12) working with primary care physicians to create a more uniform approach to counseling on obesity. CONCLUSIONS: To become more responsive to the problem of childhood obesity, WIC should consider the following: (1) providing staff training in counseling skills that educate parents on child development and child-rearing and that elicit the client's social context and personal goals, (2) shifting time allocation and programmatic emphasis in the WIC visits away from nutritional risk assessment and toward counseling, and (3) developing collaborations with primary health care providers and community agencies that impact childhood obesity. PMID- 12090834 TI - If we prescribe it, will it come? Access to asthma equipment for Medicaid-insured children and adults in the Bronx, NY. AB - CONTEXT: Asthma is a major cause of morbidity in the United States. Self management of asthma requires access to appropriate equipment. Clinical experience in an inner-city practice suggests that families encounter difficulties in filling prescriptions for spacers/holding chambers, peak flow (PF) meters, and nebulizer machines. OBJECTIVES: To determine whether Bronx, NY, pharmacies (1) carry spacers/holding chambers, PF meters, and nebulizer machines; (2) accept Medicaid insurance for them; and (3) perceive barriers to reimbursement by Medicaid for this equipment. DESIGN AND SETTING: Structured telephone survey of 100 Bronx pharmacies randomly selected from the 1999 telephone directory. PARTICIPANTS: Ninety-eight pharmacists and 2 pharmacy technicians in 100 different pharmacies. MAIN OUTCOME MEASURES: Pharmacists' reports of equipment availability, Medicaid acceptance, and reasons for not carrying equipment or accepting Medicaid. RESULTS: Overall equipment availability was as follows: spacers (68%), spacers with masks (57%), adult PF meters (40%), child-range PF meters (24%), and nebulizer machines (56%). For Medicaid recipients, equipment was less available: spacers (45%), spacers with masks (35%), adult PF meters (27%), child-range PF meters (17%), and nebulizer machines (33%). Surveyed pharmacists reported misconceptions about requirements for Medicaid reimbursement, which included the following: that Durable Medical Equipment permits are required (64% spacers and 33% PF meters), that special forms are needed (17% PF meters), or that this equipment is not covered by Medicaid (14% spacers and 8% PF meters). Of the 100 surveyed pharmacists, 32 reported difficulties with Medicaid reimbursement and 41 had never tried to receive reimbursement. CONCLUSIONS: These results suggest that (1) access to spacers/holding chambers, PF meters, and nebulizers for Medicaid-insured families is severely limited in Bronx pharmacies; (2) misunderstandings regarding Medicaid reimbursement policies are common; and (3) interventions to increase the proportion of pharmacies that dispense equipment are needed. PMID- 12090835 TI - Welfare reform and the health of young children: a sentinel survey in 6 US cities. AB - CONTEXT: Welfare reform under the 1996 Personal Responsibility and Work Opportunity Reconciliation Act replaced entitlement to cash assistance for low income families with Temporary Assistance to Needy Families, thereby terminating or decreasing cash support for many participants. Proponents anticipated that continued receipt of food stamps would offset the effects of cash benefit losses, although access to food stamps was also restricted. OBJECTIVE: To examine associations of loss or reduction of welfare with food security and health outcomes among children aged 36 months or younger at 6 urban hospitals and clinics. DESIGN AND SETTING: A multisite retrospective cohort study with cross sectional surveys at urban medical centers in 5 states and Washington, DC, from August 1998 through December 2000. PARTICIPANTS: The caregivers of 2718 children aged 36 months or younger whose households received welfare or had lost welfare through sanctions were interviewed at hospital clinics and emergency departments. MAIN OUTCOME MEASURES: Household food security status, history of hospitalization, and, for a subsample interviewed in emergency departments, whether the child was admitted to the hospital the day of the visit. RESULTS: After controlling for potential confounding factors, children in families whose welfare was terminated or reduced by sanctions had greater odds of being food insecure (adjusted odds ratio [AOR], 1.5; 95% confidence interval [CI], 1.1-1.9), of having been hospitalized since birth (AOR, 1.3; 95% CI, 1.0-1.7) and, for the emergency department subsample, of being admitted the day of an emergency department visit (AOR, 1.9; 95% CI, 1.2-3.0) compared with those without decreased benefits. Children in families whose welfare benefits were decreased administratively because of changes in income or expenses had greater odds of being food insecure (AOR, 1.5; 95% CI, 1.1-2.2) and of being admitted the day of an emergency department visit (AOR, 2.8; 95% CI, 1.4-5.6). Receiving food stamps does not mitigate the effects of the loss or reduction of welfare benefits on food security or hospitalizations. CONCLUSION: Terminating or reducing welfare benefits by sanctions, or decreasing benefits because of changes in income or expenses, is associated with greater odds that young children will experience food insecurity and hospitalizations. PMID- 12090836 TI - Mechanisms behind the failure of residents' longitudinal primary care to promote disclosure and discussion of psychosocial issues. AB - CONTEXT: Longitudinality (care by a single physician over time) and continuity (receipt of most care from a single physician) are believed to enhance patient physician relationships and facilitate disclosure of emotional distress, but some studies suggest this potential goes unrealized. OBJECTIVES: To determine whether care in a pediatric residents' continuity clinic promotes, over time, increased discussion, disclosure, and detection of parents' social and emotional distress and to understand physicians' communication behaviors underlying changes with time. DESIGN: Longitudinal, observational study of parent-physician interaction over the course of 1 year. PARTICIPANTS: One hundred ninety parents (90% African American) and their infants' primary care physicians (31 [4 Asians and 27 whites] first- and second-year pediatric residents). MAIN OUTCOME MEASURES: Frequency with which parents and physicians raised topics related to parental mood and family or social functioning; proportion of distressed parents discussing mood or functioning; and physicians' detection of parent distress. RESULTS: Physician initiation of psychosocial topics fell in the course of longitudinal relationships (odds of initiation in visits > or =6 vs odds of initiation in visits 1-5 = 0.46 [95% confidence limits, 0.31%, 0.67%]); parent initiation did not change over time nor was it increased by greater levels of continuity. Length of relationship was not associated with increased physician detection of parental distress or with increased rates of disclosure by distressed parents. Physicians' positively framed leading questions, and their avoidant responses to prior parental disclosures were significantly associated with decreased odds of problem disclosure. In contrast, visits in which parents or physicians raised psychosocial topics were characterized, on average, by 40% higher levels of physicians' "patient-centeredness" (increases of about 100 utterances per visit [95% confidence limits, 65.7%, 133.9%]). CONCLUSIONS: Longitudinal relationships between residents and patients may not be sufficient to promote the discussion, disclosure, and detection of psychosocial issues. Training in communication skills may help residents achieve the potential and goals of longitudinal care. PMID- 12090838 TI - Decreased growth during therapy with selective serotonin reuptake inhibitors. AB - BACKGROUND: There is no information on the effects of selective serotonin reuptake inhibitors (SSRIs) on growth and puberty in children. We examined growth and growth hormone secretion in 4 children treated with SSRIs for various psychiatric disorders. DESIGN: Case study. PARTICIPANTS: Four children (3 boys) aged 11.6 to 13.7 years with obsessive-compulsive disorder or Tourette syndrome. MAIN OUTCOME MEASURES: Growth, pubertal progression, and hypothalamic pituitary function. METHODS: The patients were treated with SSRIs for 6 months to 5 years (dosage, 20-100 mg/d). All were regularly examined for changes in height and bone age and for pubertal progression. They also underwent evaluation of somatotrophic axis and hypothalamic-pituitary axis function. RESULTS: All 4 patients had growth attenuation. Three of them exhibited growth retardation at a pubertal stage when a growth spurt was anticipated. Three had a decreased growth hormone response to clonidine hydrochloride stimulation and 2 to both clonidine and glucagon stimulation, and 1 had decreased 24-hour secretion of growth hormone that normalized when therapy was stopped. The rest of the endocrine evaluations were within reference ranges in all patients. At follow-up, 2 patients were being treated with somatropin while continuing SSRI therapy, and the other 2 resumed normal growth after discontinuation of therapy. CONCLUSIONS: A decrease in growth rate, possibly secondary to suppression of growth hormone secretion, may occur during SSRI therapy. As the use of this group of drugs is expected to increase in the young age groups, larger studies are warranted to investigate their effect on growth and growth hormone secretion. PMID- 12090837 TI - Teaching Spanish to pediatric emergency physicians: effects on patient satisfaction. AB - BACKGROUND: Language barriers are known to negatively affect patient satisfaction. OBJECTIVE: To determine whether a course of instruction in medical Spanish for pediatric emergency department (ED) physicians is associated with an increase in satisfaction for Spanish-speaking-only families. DESIGN, SETTING, PARTICIPANTS, AND INTERVENTION: Nine pediatric ED physicians completed a 10-week medical Spanish course. Mock clinical scenarios and testing were used to establish an improvement in each physician's ability to communicate with Spanish speaking-only families. Before (preintervention period) and after (postintervention period) the course, Spanish-speaking-only families cared for by these physicians completed satisfaction questionnaires. Professional interpreters were equally available during both the preintervention and postintervention periods. MAIN OUTCOME MEASURES: Responses to patient family satisfaction questionnaires. RESULTS: A total of 143 Spanish-speaking-only families completed satisfaction questionnaires. Preintervention (n = 85) and postintervention (n = 58) cohorts did not differ significantly in age, vital signs, length of ED visit, discharge diagnosis, or self-reported English proficiency. Physicians used a professional interpreter less often in the postintervention period (odds ratio [OR], 0.34; 95% confidence interval [CI], 0.16-0.71). Postintervention families were significantly more likely to strongly agree that "the physician was concerned about my child" (OR, 2.1; 95% CI, 1.0-4.2), "made me feel comfortable" (OR, 2.6; 95% CI, 1.1-4.4), "was respectful" (OR, 3.0; 95% CI, 1.4-6.5), and "listened to what I said" (OR, 2.9; 95% CI, 1.4-5.9). CONCLUSIONS: A 10-week medical Spanish course for pediatric ED physicians was associated with decreased interpreter use and increased family satisfaction. PMID- 12090839 TI - Trends in pediatric hospitalizations of children in washington state by insurance and chronic condition status, 1991-1998. AB - OBJECTIVE: To examine the possible impact of changes in the organization and management of the Medicaid program on hospitalization patterns for children with chronic and nonchronic conditions between January 1, 1991, and December 31, 1998. DESIGN: Longitudinal retrospective study of hospitalization patterns of children in 4 strata: Medicaid, non-Medicaid, chronic conditions, and nonchronic conditions. SETTING: Washington State. PATIENTS: Hospital discharge abstract records for all children aged 0 to 17 years profiled into those with and without a chronic condition, Medicaid, and non-Medicaid using a diagnosis-based classification system. MAIN OUTCOME MEASURES: Hospitalization and multiple hospitalization rates and length of hospital stay. RESULTS: In 1991, hospitalization and multiple hospitalization rates were higher for all Medicaid vs non-Medicaid children. From 1991 to 1998, there was a decrease in the hospitalization and multiple hospitalization rates for Medicaid children only. By 1998, rates for Medicaid children approximated those for non-Medicaid children. This decrease was greater for nonchronically ill children than for chronically ill children. Total hospitalizations in Medicaid children decreased by 4.5%. The mean length of stay in 1991 for all Medicaid hospitalized children was higher than that for non-Medicaid children (6.1 vs 5.1 days). By 1998, the length of stay decreased for both groups (5.7 vs 4.9 days). CONCLUSION: The declines in hospitalization and multiple hospitalization rates observed in Washington State Medicaid children from 1991 to 1998 may be the result of many statewide efforts to increase access and improve management for this population. PMID- 12090840 TI - Relationship between early primary care and emergency department use in early infancy by the medicaid population. AB - OBJECTIVE: To examine the relationship between the use and type of primary care and visits to the emergency department (ED) in early infancy by healthy infants who are Medicaid recipients. DESIGN: A population-based cohort study using a database linking birth certificate data to Medicaid claims. PARTICIPANTS: A total of 151 464 full-term infants born in Ohio to mothers receiving Medicaid from July 1, 1991, through June 30, 1998. MAIN OUTCOME MEASURES: The primary outcome of interest was the occurrence of an ED visit within 91 days of the neonate's birth. Bivariate and multivariate analyses were performed to determine the effect of early linkage with primary care (within 21 days of birth) on ED use in early infancy. RESULTS: Only 53% of the infants had a documented primary care visit within 21 days of birth. Twenty-eight percent of infants had at least 1 ED visit within 91 days of birth and 9% had more than 1 visit. The mean age of the neonate at the first ED visit was 39.7 days. Fifteen percent of primary care visits within 21 days of birth occurred at a hospital-based primary care clinic. After adjusting for maternal, infant, and residency characteristics and temporal differences, early primary care linkage was associated with a 16% increase in the likelihood of ED use. When the primary care visit occurred in a hospital-based primary care clinic, it was associated with a 27% increase in the likelihood of ED use. CONCLUSION: Contrary to our expectations, early primary care linkage did not result in a decreased risk of ED use. PMID- 12090841 TI - Changing nursery practice gets inner-city infants in the supine position for sleep. AB - OBJECTIVE: To determine whether an educational intervention to change nursery practice would result in more inner-city parents placing their infants in the supine position for sleep. DESIGN: We conducted semistructured interviews at the 2-week health supervision visit with 1 convenience sample of parents before and a different convenience sample of parents after an educational intervention was conducted to change nursery practice in positioning infants for sleep. SETTING: University hospital clinic located in an urban setting. PARTICIPANTS: Parents of 2-week-old infants at their first health supervision visit in an urban, university-affiliated clinic. All parents who were approached agreed to participate. INTERVENTION: Nurses were instructed to place infants exclusively in the supine position in the nursery and to instruct parents to exclusively place infants in the supine sleeping position at home. MAIN OUTCOME MEASURE: The usual sleeping position in which parents reported placing their 2-week old infants. RESULTS: Before the intervention, 41% of parents reported that a clinician had told them to place their infants to sleep in the supine position compared with 81% after the intervention (odds ratio [OR], 6.1; 95% confidence interval [CI], 3.1-12.3). Before the intervention, 37% of parents reported that the nursery staff placed their infants to sleep in the supine position, compared with 88% after the intervention (OR, 12.5; 95% CI, 5.7-27.7). Before the intervention, 42% of parents reported that they usually placed their infants to sleep in the supine position at home compared with 75% after the intervention (OR, 4.2; 95% CI, 2.1 7.9). CONCLUSION: After an educational intervention to change practice in a well newborn nursery, many more parents reported placing their infants in the supine position for sleep, which suggests that such an intervention may have an impact on the position in which parents place their children to sleep. PMID- 12090842 TI - Mental health service use in a community head start population. AB - BACKGROUND: Evaluating access to and delivery of mental health services for young children was a primary objective of the national research demonstration program Starting Early Starting Smart (SESS). OBJECTIVE: To present preliminary findings on family mental health and use of services in a community Head Start population at time of entry into a longitudinal study as part of the SESS program. SUBJECTS: Children enrolled for Head Start entry in 1998 and 1999 (N = 290; mean age, 4.3 years). Of these children, 52% were boys. METHODS: Data on demographic factors, child and parent mental health, and service use were collected from the sample at baseline. Information was gathered from primary caregivers and teachers using standardized questionnaires and structured interviews. RESULTS: There was low concordance between parent and teacher ratings of child behavior. Factors predicting behavior problems in young children varied according to whether the parent or teacher rated the child as having behavior problems. Sex (male) and home environment were associated with teachers rating the child as having a behavior problem. Parent mental health problems and problems in the parent-child relationship were associated with parent ratings. Only home environment was associated with child-focused service utilization (services that help parents manage children's behavior). CONCLUSIONS: Demographic risk factors were not associated with child behavior problems or use of mental health services in this group of Head Start children. Findings suggest that children with behavioral problems have unmet mental health service needs. Interventions designed to address both parent mental health needs and sensitivity to the developmental needs of children may increase child-focused mental health service utilization. PMID- 12090843 TI - The need for sickle cell screening among pediatric latino immigrants. PMID- 12090845 TI - Limitations in measuring asthma disparities. PMID- 12090848 TI - A piece of my mind: skydiving. PMID- 12090850 TI - Walking in beauty at Sage Memorial Hospital. PMID- 12090851 TI - TB picture brightening, but dark spots remain. PMID- 12090852 TI - From the Food and Drug Administration. PMID- 12090856 TI - Raloxifene and risk of cardiovascular events. PMID- 12090858 TI - Diagnosis and treatment of cutaneous anthrax. PMID- 12090859 TI - Diagnosis and treatment of cutaneous anthrax. PMID- 12090861 TI - Camphor intoxication after cao gio (coin rubbing). PMID- 12090862 TI - Cardiovascular disease outcomes during 6.8 years of hormone therapy: Heart and Estrogen/progestin Replacement Study follow-up (HERS II). AB - CONTEXT: The Heart and Estrogen/progestin Replacement Study (HERS) found no overall reduction in risk of coronary heart disease (CHD) events among postmenopausal women with CHD. However, in the hormone group, findings did suggest a higher risk of CHD events during the first year, and a decreased risk during years 3 to 5. OBJECTIVE: To determine if the risk reduction observed in the later years of HERS persisted and resulted in an overall reduced risk of CHD events with additional years of follow-up. DESIGN AND SETTING: Randomized, blinded, placebo-controlled trial of 4.1 years' duration (HERS) and subsequent unblinded follow-up for 2.7 years (HERS II) conducted at outpatient and community settings at 20 US clinical centers. PARTICIPANTS: A total of 2763 postmenopausal women with CHD and average age of 67 years at enrollment in HERS; 2321 women (93% of those surviving) consented to follow-up in HERS II. INTERVENTION: Participants were randomly assigned to receive 0.625 mg/d of conjugated estrogens and 2.5 mg of medroxyprogesterone acetate (n = 1380), or placebo (n = 1383) during HERS; open-label hormone therapy was prescribed at personal physicians' discretion during HERS II. The proportions with at least 80% adherence to hormones declined from 81% (year 1) to 45% (year 6) in the hormone group, and increased from 0% (year 1) to 8% (year 6) in the placebo group. MAIN OUTCOME MEASURES: The primary outcome was nonfatal myocardial infarction and CHD death. Secondary cardiovascular events were coronary revascularization, hospitalization for unstable angina or congestive heart failure, nonfatal ventricular arrhythmia, sudden death, stroke or transient ischemic attack, and peripheral arterial disease. RESULTS: There were no significant decreases in rates of primary CHD events or secondary cardiovascular events among women assigned to the hormone group compared with the placebo group in HERS, HERS II, or overall. The unadjusted relative hazard (RH) for CHD events in HERS was 0.99 (95% confidence interval [CI], 0.81-1.22); HERS II, 1.00 (95% CI, 0.77-1.29); and overall, 0.99 (0.84-1.17). The overall RHs were similar after adjustment for potential confounders and differential use of statins between treatment groups (RH, 0.97; 95% CI, 0.82-1.14), and in analyses restricted to women who were adherent to randomized treatment assignment (RH, 0.96; 95% CI, 0.77-1.19). CONCLUSIONS: Lower rates of CHD events among women in the hormone group in the final years of HERS did not persist during additional years of follow-up. After 6.8 years, hormone therapy did not reduce risk of cardiovascular events in women with CHD. Postmenopausal hormone therapy should not be used to reduce risk for CHD events in women with CHD. PMID- 12090863 TI - Noncardiovascular disease outcomes during 6.8 years of hormone therapy: Heart and Estrogen/progestin Replacement Study follow-up (HERS II). AB - CONTEXT: The Heart and Estrogen/progestin Replacement Study (HERS) was a randomized trial of estrogen plus progestin therapy after menopause. OBJECTIVE: To examine the effect of long-term postmenopausal hormone therapy on common noncardiovascular disease outcomes. DESIGN AND SETTING: Randomized, blinded, placebo-controlled trial of 4.1 years' duration (HERS) and subsequent open-label observational follow-up for 2.7 years (HERS II), carried out between 1993 and 2000 in outpatient and community settings at 20 US clinical centers. PARTICIPANTS: A total of 2763 postmenopausal women with coronary disease and average age of 67 years at enrollment in HERS; 2321 women (93% of those surviving) consented to follow-up in HERS II. INTERVENTION: Participants were randomly assigned to receive 0.625 mg/d of conjugated estrogens plus 2.5 mg of medroxyprogesterone acetate (n = 1380) or placebo (n = 1383) during HERS; open label hormone therapy was prescribed at personal physicians' discretion during HERS II. The proportions with at least 80% adherence to hormones declined from 81% (year 1) to 45% (year 6) in the hormone group and increased from 0% (year 1) to 8% (year 6) in the placebo group. MAIN OUTCOME MEASURES: Thromboembolic events, biliary tract surgery, cancer, fracture, and total mortality. RESULTS: Comparing women assigned to hormone therapy with those assigned to placebo, the unadjusted intention-to-treat relative hazard (RH) for venous thromboembolism declined from 2.66 (95% confidence interval [CI], 1.41-5.04) during HERS to 1.40 (95% CI, 0.64-3.05) during HERS II (P for time trend =.08); it was 2.08 overall for the 6.8 years (95% CI, 1.28-3.40), and 3 of the 73 women with thromboembolism died within 30 days due to pulmonary embolism. The overall RH for biliary tract surgery was 1.48 (95% CI, 1.12-1.95); for any cancer, 1.19 (95% CI, 0.95-1.50); and for any fracture, 1.04 (95% CI, 0.87-1.25). There were 261 deaths among those assigned to hormone therapy and 239 among those assigned to placebo (RH, 1.10; 95% CI, 0.92-1.31). Adjusted and as-treated analyses did not alter our conclusions. CONCLUSIONS: Treatment for 6.8 years with estrogen plus progestin in older women with coronary disease increased the rates of venous thromboembolism and biliary tract surgery. Trends in other disease outcomes were not favorable and should be assessed in larger trials and in broader populations. PMID- 12090864 TI - Cerebral white matter lesions, retinopathy, and incident clinical stroke. AB - CONTEXT: White matter lesions (WMLs) detected on cerebral imaging scans have been hypothesized to have a microvascular etiology and to precede the development of clinical stroke. However, few clinical data are available to support these hypotheses. OBJECTIVE: To examine the relationship of WMLs, retinal microvascular abnormalities, and incident clinical stroke in healthy, middle-aged men and women. DESIGN AND SETTING: The Atherosclerosis Risk in Communities Study (ARIC), a prospective, population-based cohort study conducted in 4 US communities and initiated in 1987-1989. PARTICIPANTS: A total of 1684 persons aged 51 to 72 years who had cerebral magnetic resonance imaging (MRI) and retinal photography at the third examination (1993-1995). MAIN OUTCOME MEASURES: Odds of WMLs, defined by standardized methods from MRI, by presence or absence of specific retinal microvascular abnormality (eg, microaneurysm, retinal hemorrhage) on retinal photograph; incident clinical stroke, ascertained after a median follow-up of 4.7 years, according to presence or absence of WMLs and retinopathy. RESULTS: Persons with retinopathy were more likely to have WMLs than those without retinopathy (22.9% vs 9.9%; odds ratio, 2.5; 95% confidence interval [CI], 1.5-4.0, adjusted for age, sex, race, and vascular risk factors). The 5-year cumulative incidence of clinical stroke was higher in persons with vs without WMLs (6.8% vs 1.4%; adjusted relative risk [RR], 3.4; 95% CI, 1.5-7.7) and in persons with vs without retinopathy (8.0% vs 1.4%; adjusted RR, 4.9; 95% CI, 2.0-11.9). Persons with both WMLs and retinopathy had a significantly higher 5-year cumulative incidence of stroke than those without either WMLs or retinopathy (20.0% vs 1.4%; adjusted RR, 18.1; 95% CI, 5.9-55.4). CONCLUSIONS: In this cohort, middle-aged persons with cerebral WMLs detected on MRI were more likely to have retinal microvascular abnormalities and to have an increased risk of clinical stroke than people without WMLs. The risk of stroke was higher when retinopathy was simultaneously present in persons with WMLs. PMID- 12090865 TI - Changing profile of infective endocarditis: results of a 1-year survey in France. AB - CONTEXT: Since the first modern clinical description of infective endocarditis (IE) at the end of the 19th century, the profile of the disease has evolved continuously, as highlighted in epidemiological studies including a French survey performed in 1991. OBJECTIVE: To update information gained from the 1991 study on the epidemiology of IE in France. DESIGN AND SETTING: Population-based survey conducted from January through December 1999 in all hospitals in 6 French regions representing 26% of the population (16 million inhabitants). PATIENTS: Three hundred ninety adult inpatients diagnosed with IE according to Duke criteria. MAIN OUTCOME MEASURES: Incidence of IE; proportion of patients with underlying heart disease; clinical characteristics; causative microorganisms; surgical and mortality outcomes. RESULTS: The annual age- and sex-standardized incidence was 31 (95% confidence interval [CI], 28-35) cases per million, not including the region of New Caledonia, which had 161 (95% CI, 117-216) cases per million. There was no previously known heart disease in 47% of the cases. The proportion of prosthetic-valve IE was 16%. Causative microorganisms were: streptococci, 48% (group D streptococci, 25%; oral streptococci, 17%, pyogenic streptococci, 6%); enterococci, 8%; Abiotrophia species, 2%; staphylococci, 29%; and other or multiple pathogens, 8%. Blood cultures were negative in 9% and no microorganism was identified in 5% of the cases. Early valve surgery was performed in 49% of the patients. In-hospital mortality was 16%. Compared with 1991, this study showed a decreased incidence of IE in patients with previously known underlying heart disease (20.6 cases per million vs 15.1 cases per million; P<.001); a smaller incidence of oral streptococcal IE (7.8 cases per million vs 5.1 cases per million; P<.001), compensated by a larger proportion of IE due to group D streptococci (5.3 cases per million vs 6.2 cases per million; P =.67) and staphylococci (4.9 cases per million vs 5.7 cases per million; P =.97); an increased rate of early valve surgery (31.2% vs 49.7%; P<.001); and a decreased in-hospital mortality rate (21.6% vs 16.6%; P =.08). CONCLUSION: Although the incidence of IE has not changed, important changes in disease characteristics, treatment, and outcomes were noted. PMID- 12090867 TI - Responding to requests for physician-assisted suicide: "These are uncharted waters for both of us...". AB - Studies of dying patients have shown that about half would like the option of physician-assisted suicide (PAS) to be available for possible future use. Those percentages decrease significantly with each step patients take toward action. Studies show that although about 10% of patients seriously consider PAS, only 1% of dying patients specifically request it, and 1 in 10 of those patients actually receive and take a lethal prescription. However, most patients' desires for PAS diminish as their underlying concerns are identified and addressed directly. To help identify concerns motivating a patient's request for PAS, physicians should talk with patients about their expectations and fears, options for end-of-life care, goals, family concerns and burdens, suffering or physical symptoms, sense of meaning and quality of life, and symptoms of depression. A patient with advanced amyotrophic lateral sclerosis (ALS) who requested PAS illustrates how a hasty response may adversely affect patient care and the health care team. Although physicians should remain mindful of their personal, moral, and legal concerns, these concerns should not override their willingness to explore what motivates a patient to make this request. When this approach is taken, suffering can be optimally alleviated and, in almost all cases, the patient's wishes can be met without PAS. PMID- 12090866 TI - The health of Latino children: urgent priorities, unanswered questions, and a research agenda. AB - Latinos recently became the largest racial/ethnic minority group of US children. The Latino Consortium of the American Academy of Pediatrics Center for Child Health Research, consisting of 13 expert panelists, identified the most important urgent priorities and unanswered questions in Latino child health. Conclusions were drawn when consensus was reached among members, with refinement through multiple iterations. A consensus statement with supporting references was drafted and revised. This article summarizes the key issues, including lack of validated research instruments, frequent unjustified exclusion from studies, and failure to analyze data by pertinent subgroups. Latino children are at high risk for behavioral and developmental disorders, and there are many unanswered questions about their mental health needs and use of services. The prevalence of dental caries is disproportionately higher for Latino children, but the reasons for this disparity are unclear. Culture and language can profoundly affect Latino children's health, but not enough cultural competency training of health care professionals and provision of linguistically appropriate care occur. Latinos are underrepresented at every level of the health care professions. Latino children are at high risk for school dropout, environmental hazards, obesity, diabetes mellitus, asthma, lack of health insurance, nonfinancial barriers to health care access, and impaired quality of care, but many key questions in these areas remain unanswered. This article suggests areas in which more research is needed and ways to improve research and care of Latino children. PMID- 12090868 TI - Hormone replacement therapy for prevention: more evidence, more pessimism. PMID- 12090874 TI - Research on stored biological samples is still research. PMID- 12090875 TI - Controversies in internal medicine: a new feature in the ARCHIVES. PMID- 12090876 TI - Lack of evidence-based research for idiopathic low back pain: the importance of a specific diagnosis. PMID- 12090877 TI - Diagnostic evaluation of LBP: reaching a specific diagnosis is often impossible. PMID- 12090880 TI - Preoperative or postoperative start of prophylaxis for venous thromboembolism with low-molecular-weight heparin in elective hip surgery? AB - BACKGROUND: Prophylaxis of venous thromboembolism with low-molecular-weight heparins in patients undergoing major orthopedic surgery is currently initiated according to at least 3 different regimens. In Europe, traditionally, prophylaxis is started 12 hours before surgery, whereas in North America it is initiated 12 to 48 hours postoperatively. The third regimen (perioperative) begins prophylaxis either earlier than 12 hours before or 12 hours after surgery. Unfortunately, the optimal regimen is uncertain because direct comparisons among these regimens with sufficiently large sample sizes are not available. OBJECTIVE: To assess, in a systematic review, the relative efficacy and safety of the 3 low-molecular-weight heparin regimens used to prevent thrombosis after total hip replacement. The incidence of postoperative thrombosis detected by contrast venography was used as the measure of efficacy and the rate of major bleeding was used as the measure of safety. METHODS: We pooled the results of all published studies, which met the following criteria: (1) inclusion of in at least 1 arm of the study of a dose of low-molecular-weight heparin that is approved for both preoperative and postoperative initiation of prophylaxis; (2) the use of mandatory bilateral contrast venography, performed between days 6 and 15 postoperatively; (3) thromboprophylaxis continued until venography; (4) independent reading of venograms; and (5) assessment of clinically overt major bleeding by predefined criteria. Articles were excluded if no separate data could be obtained for patients undergoing elective hip surgery (in case of patient mix), or if they were reported more than once. RESULTS: In the 1926 patients who used a preoperative regimen, the incidence of postoperative deep vein thrombosis was 19.2% (95% confidence interval [CI], 17%-21%). In the cohort of 925 patients who received a perioperative regimen, the rate of deep vein thrombosis was 12.4% (95% CI, 10%-14%), whereas in the group of 694 patients who received a postoperative regimen, it was 14.4% (95% CI, 12%-17%). The rate of major bleeding was 1.4% (95% CI, 1%-2%) in the preoperative group, 6.3% (95% CI, 5%-7%) in the perioperative group, and 2.5% (95% CI, 1%-3%) in the postoperative group. CONCLUSIONS: We find no convincing evidence that starting prophylaxis preoperatively is associated with a lower incidence of venous thromboembolism than starting postoperatively. Perioperative regimens may lower the risk of postoperative thrombosis, but if so, this positive effect is offset by an increase in postoperative major bleeding. PMID- 12090881 TI - The debate over research on stored biological samples: what do sources think? AB - BACKGROUND: The debate over informed consent for research on stored biological samples has enormous scientific implications. Unfortunately, there are no data on individuals' attitudes regarding when their consent should be obtained for such research. METHODS: Data were gathered using a telephone survey of 504 individuals living in the United States. Two cohorts were studied: (1) individuals who had participated in clinical research and contributed biological samples and (2) randomly selected Medicare recipients. RESULTS: Of the respondents, 65.8% would require their consent for research on clinically derived, personally identified samples; 27.3% would require it for research on clinically derived samples that are "anonymized." For research-derived samples, 29.0% of the respondents would require their consent if the samples retain personal identifiers; 12.1% would require it if the samples are anonymized before the research is conducted. Also, 88.8% would want to be informed of results of uncertain clinical significance, and 91.9% would not impose greater safeguards on future research on a different disease. CONCLUSIONS: Current practice and policy recommendations regarding research using stored biological samples may be inconsistent with sources' preferences in several respects. In particular, it appears that most sources want to control whether their samples are used for research purposes, are not concerned with the particular disease that will be studied, and want to receive results of uncertain clinical significance. Follow-up research will be needed to assess the generalizability of the current data. PMID- 12090882 TI - Short-duration prophylaxis against venous thromboembolism after total hip or knee replacement: a meta-analysis of prospective studies investigating symptomatic outcomes. AB - BACKGROUND: The prevalence of asymptomatic deep vein thrombosis diagnosed by venography after hip or knee replacement remains high despite 7 to 10 days of anticoagulant prophylaxis. However, the risk of symptomatic events in such patients is unclear. We performed a meta-analysis to provide reliable estimates of the risk of symptomatic venous thromboembolism occurring within 3 months of hip or knee replacement in patients who received short-duration (7-10 days) anticoagulant prophylaxis. METHODS: The MEDLINE, EMBASE, and Cochrane databases were searched from January 1993 to March 2001, supplemented by a manual search of bibliographies and conference abstracts, to identify prospective studies of patients undergoing hip or knee replacement who received short-duration prophylaxis (ie, 7-10 days of fixed-dose low-molecular-weight heparin or adjusted dose warfarin, with a target international normalized ratio of 2.0-3.0). Studies were classified as clinical outcome studies if the outcome was symptomatic venous thromboembolism or as venographic outcome studies if the outcome was asymptomatic deep vein thrombosis diagnosed after bilateral venography. RESULTS: There were 4 clinical outcome studies with 6089 patients who had 3 months of follow-up, and 13 venographic outcome studies with 7080 patients who had venography 7 to 10 days after surgery. In clinical outcome studies, the 3-month incidence of nonfatal venous thromboembolism was 3.2% (95% confidence interval [CI], 2.0%-4.4%), and the 3-month incidence of fatal pulmonary embolism was 0.10% (95% CI, 0.02% 0.20%). The postprophylaxis incidence of nonfatal venous thromboembolism was 2.2% (95% CI, 1.4%-3.0%), and the incidence of fatal pulmonary embolism was 0.05% (95% CI, 0%-0.12%). The postprophylaxis incidence of symptomatic venous thromboembolism was higher after hip than after knee replacement (2.5% vs 1.4%; P=.02). In venographic outcome studies, the prevalence of deep vein thrombosis (total and proximal) was higher after knee than after hip replacement (total: 38.8% vs 16.4%; P<.001; proximal: 7.6% vs 3.8%; P<.001). CONCLUSIONS: In patients who undergo hip or knee replacement and receive short-duration anticoagulant prophylaxis, symptomatic nonfatal venous thromboembolism will occur in about 1 of 32 patients and fatal pulmonary embolism will occur in about 1 of 1000 patients within 3 months of the surgery. Although the prevalence of asymptomatic deep vein thrombosis is more than 2-fold higher after knee replacement than after hip replacement 7 to 10 days after surgery, in the subsequent 3 months, symptomatic venous thromboembolism is more likely to occur after hip replacement. PMID- 12090883 TI - Vitamin supplement use in a low-risk population of US male physicians and subsequent cardiovascular mortality. AB - BACKGROUND: Although basic research suggests that vitamins may have an important role in the prevention of cardiovascular diseases (CVD), the data from cohort studies and clinical trials are inconclusive. METHODS: This prospective cohort study was conducted among 83 639 male physicians residing in the United States who had no history of CVD or cancer. At baseline, data on use of vitamin E, ascorbic acid (vitamin C), and multivitamin supplements were provided by a self administered questionnaire. Mortality from CVD and coronary heart disease (CHD) was assessed by death certificate review. RESULTS: Use of supplements was reported by 29% of the participants. During a mean follow-up of 5.5 years, 1037 CVD deaths occurred, including 608 CHD deaths. After adjustment for several cardiovascular risk factors, supplement use was not significantly associated with total CVD or CHD mortality. For vitamin E use, the relative risks (RRs) were 0.92 (95% confidence interval [CI], 0.70-1.21) for total CVD mortality and 0.88 (95% CI, 0.61-1.27) for CHD mortality; for use of vitamin C, the RRs were 0.88 (95% CI, 0.70-1.12) for total CVD mortality and 0.86 (95% CI, 0.63-1.18) for CHD mortality; and for use of multivitamin supplements, the RRs were 1.07 (95% CI, 0.91-1.25) for total CVD mortality and 1.02 (95% CI, 0.83-1.25) for CHD mortality. CONCLUSIONS: In this large cohort of apparently healthy US male physicians, self-selected supplementation with vitamin E, vitamin C, or multivitamins was not associated with a significant decrease in total CVD or CHD mortality. Data from ongoing large randomized trials will be necessary to definitely establish small potential benefits of vitamin supplements on subsequent cardiovascular risk. PMID- 12090884 TI - Obesity is a risk factor for dyspnea but not for airflow obstruction. AB - BACKGROUND: Previous research suggests that obesity is an important risk factor for asthma. However, since obesity can cause dyspnea through mechanisms other than airflow obstruction, diagnostic misclassification of asthma could partially account for this association. OBJECTIVE: To determine whether there is a relationship between obesity and airflow obstruction. METHODS: A total of 16 171 participants (17 years or older) from the Third National Health and Nutrition Examination Survey (NHANES III) were divided into 5 quintiles based on their body mass index (BMI) to determine the association between BMI quintile and risk of self-reported asthma, bronchodilator use, exercise performance, and airflow obstruction. Significant airflow obstruction was defined as a ratio less than 80% the predicted value of forced expiratory volume in 1 second to forced vital capacity adjusted for age, sex, and race. RESULTS: The highest BMI quintile (ie, the most obese participants) had the greatest risk of self-reported asthma (odds ratio [OR], 1.50; 95% confidence interval [CI], 1.24-1.81), bronchodilator use (OR, 1.94; 95% CI, 1.38-2.72), and dyspnea with exertion (OR, 2.66; 95% CI, 2.35 3.00). Paradoxically, the highest BMI quintile had the lowest risk for significant airflow obstruction (P =.001). CONCLUSIONS: This study demonstrates that while obesity is a risk factor for self-reported asthma, obese participants are at a lower risk for (objective) airflow obstruction. Many more obese than nonobese participants were using bronchodilators despite a lack of objective evidence for airflow obstruction. These data suggest that mechanisms other than airflow obstruction are responsible for dyspnea genesis in obesity and that asthma might be overdiagnosed in the obese population. PMID- 12090885 TI - The hospital water supply as a source of nosocomial infections: a plea for action. AB - BACKGROUND: Microbiologically contaminated drinking water is a cause of community acquired infection, and guidelines for prevention of such infections have been established. Microbes in hospital water can also cause nosocomial infection, yet guidelines for preventing such infections do not exist. The purpose of this review is to assess the magnitude of the problem caused by waterborne nosocomial infections and to plea for immediate action for their prevention. METHODS: We conducted a MEDLINE search of the literature published between January 1, 1966, and December 31, 2001. STUDY SELECTION AND DATA EXTRACTION: Investigations in which microorganisms (other than Legionella species) caused waterborne nosocomial infections and public health agency recommendations for drinking water. RESULTS: Forty-three outbreaks of waterborne nosocomial infections have been reported, and an estimated 1400 deaths occur each year in the United States as a result of waterborne nosocomial pneumonias caused by Pseudomonas aeruginosa alone. Despite the availability of effective control measures, no clear guidelines exist for the prevention of these infections. By contrast, guidelines for the prevention of community-acquired waterborne infections are now routinely used. Hospitals caring for patients at high risk for infection do not enforce the standards of water quality recommended by US and United Kingdom public health agencies for the patients' community counterparts. CONCLUSION: Because of the seriousness of these nosocomial waterborne infections and the availability, low cost, and proven effectiveness of sterile water, we recommend that hospitalized patients at high risk for infection avoid exposure to hospital water and use sterile water instead. PMID- 12090886 TI - Risk factors for urological symptoms in a cohort of users of the HIV protease inhibitor indinavir sulfate: the ATHENA cohort. AB - BACKGROUND: Nephrolithiasis is a well-known complication of indinavir treatment and may result in urological symptoms ranging from renal colic to renal insufficiency. OBJECTIVE: To obtain further knowledge regarding the incidence and risk factors of urological symptoms associated with indinavir sulfate use. METHODS: This study was performed in the ATHENA (AIDS Therapy Evaluation National AIDS Therapy Evaluation Centre) cohort of patients infected with human immunodeficiency virus (HIV) receiving antiretroviral therapy in the Netherlands. The incidence rate of urological symptoms was assessed in a subcohort of 1219 patients starting HIV protease inhibitor treatment after 1996. Urological symptoms were defined as an initial report of nephrolithiasis, renal colic, flank pain, hematuria, renal insufficiency, or nephropathy. Using multivariate Cox regression analysis, risk factors for urological symptoms during indinavir treatment were subsequently studied among the subset of 644 patients who started indinavir treatment after 1996. RESULTS: The incidence of urological symptoms was 8.3 per 100 treatment-years for indinavir vs 0.8 per 100 treatment-years for other HIV protease inhibitors. Risk factors for urological symptoms during indinavir treatment were low weight (relative risk [RR], 2.1; 95% confidence interval [CI], 1.1-3.9), low lean body mass (RR, 1.7; 95% CI, 1.0-2.9), undetectable HIV-1 RNA when starting indinavir treatment (RR, 3.2; 95% CI, 1.5 6.0), prior treatment change because of intolerance (RR, 2.4; 95% CI, 1.2-5.1), indinavir regimens of 1000 mg or more twice daily (RR, 3.1; 95% CI, 1.3-8.2), and warm environmental temperatures (RR, 3.9; 95% CI, 1.7-8.8). Risk estimates were highest among patients with a low lean body mass. CONCLUSION: Increased alertness for urological symptoms is warranted for patients starting indinavir treatment, particularly among those with a low lean body mass, during indinavir regimens of 1000 mg or more twice daily, and in warm weather environments. PMID- 12090887 TI - Effect of fracture on the health care use of nursing home residents. AB - BACKGROUND: Osteoporotic fractures result in increased health care use. Care following fracture has been characterized for community dwellers but not for nursing home residents, whose fracture rates are as much as 11 times higher than those of age-matched community dwellers. Knowing the amount of care following fracture may help determine the effects of fracture prevention on use and costs in this population. METHODS: A prospective cohort study was conducted, with 18 months of follow-up, of 1427 randomly selected white, female nursing home residents 65 years and older from 47 randomly selected nursing homes in Maryland. RESULTS: After controlling for age, comorbidities, and mobility, nursing home residents who experienced a fracture were hospitalized more than 15 times as often as those who did not in the month following the fracture (relative rate, 15.35; 95% confidence interval, 12.27-19.21) and at a higher rate from 3 through 12 months postfracture. Rates in the first month were higher for persons with a hip fracture (relative rate, 31.01; 95% confidence interval, 26.52-36.24). Rates of emergency department use and contacts with physicians and therapists were increased, the latter two for 12 months following fracture. Also, before the fracture, patients who experienced a fracture visited the emergency department and had more physician contacts; for those with a hip fracture, there were fewer prefracture hospitalizations. CONCLUSIONS: Health care use remained elevated through 1 year postfracture. Comparisons with community patients suggest that this care may be less than what would be provided in other settings. For patients who fractured a hip, higher use decreased after 6 months, similar to community cohorts. Nursing home residents who visit the emergency department may warrant special screening for a fracture. PMID- 12090888 TI - Using antineutrophil cytoplasmic antibody testing to diagnose vasculitis: can test-ordering guidelines improve diagnostic accuracy? AB - BACKGROUND: Antineutrophil cytoplasmic antibodies (ANCAs) are strongly associated with Wegener granulomatosis, Churg-Strauss angiitis, microscopic polyangiitis, and pauci-immune glomerulonephritis, referred to collectively as ANCA-associated vasculitis (AAVs). It is unclear how accurate ANCA measurement is for diagnosing AAV in diverse populations or whether proposed ANCA test-ordering guidelines improve test performance. METHODS: We assembled a retrospective case series of hospitalized and ambulatory patients from 2 academic medical centers to assess the diagnostic accuracy of ANCA measurement by enzyme-linked immunosorbent assay in identifying cases of AAV. In addition, we assessed the effect of applying proposed ANCA test-ordering guidelines on test performance. RESULTS: For ANCA testing, sensitivity was 81%; specificity, 98%; positive predictive value, 54%; and negative predictive value, 99%. There were no significant changes in operating characteristics after applying the guideline criteria. Using guidelines would have decreased ANCA test ordering by 23% and would have decreased the false positive rate by 27%. No cases of AAV would have been missed if only patients fulfilling the guidelines were ANCA tested. CONCLUSION: A positive result on an enzyme-linked immunosorbent assay ANCA test, as it is currently ordered, is not a definitive diagnostic indicator of AAV. Compliance with guidelines for ANCA testing would decrease the number of false-positive results and has the potential to reduce total test expenditures. PMID- 12090889 TI - Citywide clonal outbreak of multiresistant Acinetobacter baumannii and Pseudomonas aeruginosa in Brooklyn, NY: the preantibiotic era has returned. AB - BACKGROUND: Carbapenems are important agents for treating nosocomial gram negative infections. Carbapenem-resistant bacteria have become increasingly problematic in certain regions. This study determined the citywide prevalence and molecular epidemiological features of carbapenem-resistant Acinetobacter baumannii and Pseudomonas aeruginosa in Brooklyn, NY. METHODS: All unique patient isolates of A baumannii and P aeruginosa were collected from 15 Brooklyn hospitals from July 1, 1999, through September 30, 1999. Antibiotic susceptibilities, the genetic relatedness of resistant isolates, and the relationship between antibiotic use and resistance rates were determined. RESULTS: A total of 419 isolates of A baumannii and 823 isolates of P aeruginosa were collected. For A baumannii, 53% were resistant to meropenem and/or imipenem, and 12% were resistant to all standard antibiotics. Ribotyping revealed that a single clone accounted for 62% of the samples and was isolated from patients at all 15 hospitals. The rate of carbapenem resistance was associated with cephalosporin use at each hospital (P =.004). For P aeruginosa, 24% were resistant to imipenem, 5% to amikacin, and 15% to 29% to other antipseudomonal agents. Ribotyping revealed that 3 clones accounted for nearly half of the isolates and were shared by most hospitals. CONCLUSIONS: Approximately 400 patients were infected or colonized with carbapenem-resistant A baumannii and P aeruginosa during a 3-month period in 1999. A few strains have spread widely throughout hospitals in this region. The prevalence of resistant A baumannii seems to be correlated with cephalosporin use. Multiresistant hospital-acquired bacteria should be viewed as a serious public health issue rather than an individual hospital's problem. An intensive coordinated effort will be needed to effectively address this problem. PMID- 12090890 TI - Systemic vasculitis with lymphocytic temporal arteritis and Toxocara canis infection. AB - Vasculitis associated with helminthic infection is rare. We report a case of systemic negative antineutrophil cytoplasmic antibodies vasculitis associated with Toxocara canis infection documented by enzyme-linked immunosorbent assay and Western blot analysis. This case report is unusual because of lymphocytic temporal arteritis and renal involvement. The spontaneous remission of systemic vasculitis without use of steroids or immunosuppressive agents favors a close relationship between vasculitis and the parasite that is more than a coincidental association. To our knowledge, this is the first observation of temporal vasculitis associated with helminthic infection, extending the clinical spectrum of visceral larva migrans. PMID- 12090891 TI - Isolated office hypertension: ambulatory BP monitoring is the only answer. PMID- 12090893 TI - ABPM is valuable for the management of hypertension. PMID- 12090892 TI - Treatment of IOH and risk of visual complications. PMID- 12090896 TI - Better hypertension control? Not without broad interventions by strong public health institutions. PMID- 12090898 TI - Endogenous salicylates, aspirin, and inflammation. PMID- 12090899 TI - Understanding torsade de pointes: a bridge to tailored drug therapy? PMID- 12090900 TI - Genetic defects of cardiac ion channels. The hidden substrate for torsades de pointes. AB - The novel insights provided by the molecular genetic applied to the study of cardiac arrhythmias have just started to scratch the surface of the complex relationships between the genetic abnormalities of ion channel structure and the susceptibility to life threatening ventricular arrhythmias. These crucial pieces of information that are being gathered may have broader implications than those concerning the relatively restricted field of inherited arrhythmogenic diseases. An area of possible crucial application of the molecular genetic of cardiac ion channels, is the so-called drug-induced Torsade de Pointes (TdP) and acquired Long QT Syndrome. This condition is defined as an abnormal response to the administration of a variety of drugs which, in susceptible subjects, may induce an excessive QT interval prolongation and possibly lead to the onset of life threatening ventricular tachyarrhythmias (drug-induced TdP). The "proof of concept" that sub-clinical variants of the inherited long QT syndrome may play a causative role has been recently brought to light. However, large population based studies are still needed in order to quantify the epidemiological relevance of such findings. The future developments in this area of research will lead to the availability of pre-prescription genotyping for the identification of the susceptible subjects and to the development of safer drugs. PMID- 12090901 TI - Dispersion of repolarisation and the autonomic system-can we predict torsade de pointes? AB - Prediction of the onset of Torsade de Pointes (TdP) is a challenge for clinicians, because the list of drugs affecting myocardial repolarisation is continuously increasing. Alterations in the activity of autonomic nervous system and abnormalities in ventricular repolarisation are key features both as triggers and as markers for vulnerability to TdP. Recent molecular genetic studies have shown that autonomic nervous system has channel and gene specific influences on vulnerability to TdP. New analysis techniques in quantifying the dispersion of repolarisation have also been developed. QT interval dispersion, defined as a difference between the maximum and minimum QT interval measured from the standard 12-lead electrocardiogram (ECG), is one such method. In preliminary studies, QT dispersion has provided more accurate information on the risk for TdP than the measurement of the length of QT interval from a single ECG lead. Unfortunately, QT dispersion is entailed with some conceptual and methodological problems, which impairs its widespread clinical utility in risk stratification. Despite advances in the understanding of the role of autonomic nervous system as a trigger of TdP in specific gene mutations and improved clinical methods in detecting repolarisation abnormalities, accurate and reliable prediction of the onset of TdP still remains an unresolved clinical problem in individual cases. PMID- 12090902 TI - Physician-induced torsade de pointes--therapeutic implications. AB - "Torsade de pointes" (TdP) is a clinico-electrocardiographic syndrome characterized by an abnormally prolonged QT interval and the occurrence of potentially life-threatening ventricular tachyarrhythmias. Two mayor causes can be distinguished: congenital and acquired long QT syndrome. Whereas the former has recently been identified as an ion channelopathy, the mechanisms underlying acquired long QT syndrome are far from being understood. It has been suggested that patients with the acquired form of the disease may suffer from a clinically hidden form of the congenital variant. However, recent studies have yielded only a small number of individual cases in whom genetic analyses revealed the presence of an ion channel gene mutation. Since acquired long QT syndrome is most often induced by drugs prolonging myocardial repolarization, it is largely an iatrogenic disease. In order to prevent unwitting exposure to risk, physicians prescribing agents that may prolong repolarization need to be aware of the typical clinico-electrocardiographic characteristics of drug-induced TdP, and its diagnosis and management. A clearer delineation of the risk factors predisposing to abnormal prolongation of repolarization, and a more precise quantification of the torsadogenic potency of individual drugs appear mandatory in order to prevent or at least minimize the occurrence of this potentially fatal adverse effect of certain drugs. PMID- 12090903 TI - Effects of adrenaline on electrophysiological parameters during short exposure to global ischemia. A ventricular fibrillation study in isolated heart. AB - BACKGROUND: The mechanisms by which adrenaline may enhance defibrillation success rate, is poorly understood. OBJECTIVES: To study electrophysiological effects of adrenaline during short exposure to global ischemia. METHODS: In isolated perfused feline hearts, coronary perfusion was eliminated repeatedly for 1 min with 10 min reperfusion intervals. Treatment included: (1) continuous perfusion alone-control, (2) global ischemia alone, (3) adrenaline (10(-7) M) during perfusion, (4) adrenaline (10(-7) M) during global ischemia (n = 10), in separate hearts, (5) control and higher adrenaline concentration (1 x 10(-6) M), (6) during perfusion, (7) during global ischemia (n = 9). Measurements during pacing included: (1) diastolic threshold of excitability; (2) refractoriness; (3) epicardial conduction time; and (4) all tissue resistivity to indirectly detect changes in passive properties of conduction. Measurements during 1 min (or 90 sec) of electrically induced ventricular fibrillation included-all tissue resistivity and, based on maximal entropy spectral analysis and normalized entropy, rate of arrhythmia and degree of arrhythmia organization. RESULTS: Adrenaline (10(-7) M) during global ischemia vs control caused spontaneous arrhythmia termination, increased threshold significantly but reduced conduction time. Higher adrenaline concentration (1 x 10(-6) M) during global ischemia improved the passive properties of conduction and arrhythmia organization and reduced arrhythmia rate. Global ischemia alone increased conduction time but had a deleterious effect on passive properties. Adrenaline (10(-7) M) during perfusion improved conduction, but did so less than during global ischemia. Higher adrenaline concentration during perfusion (10(-6) M) improved arrhythmia organization and caused spontaneous arrhythmia termination but again less than during global ischemia. CONCLUSIONS: During short periods of global ischemia adrenaline improved the passive properties of conduction and arrhythmia organization, reduced arrhythmia rate and increased its spontaneous termination. Such changes may be operative in improving defibrillation success. PMID- 12090904 TI - HMG-CoA reductase inhibitors reduce interleukin-6 synthesis in human vascular smooth muscle cells. AB - Interleukin-6 (IL-6) is a key molecule in chronic inflammation and has been implicated in the progression of atherosclerosis. HMG-CoA reductase inhibitors (statins) may reduce the cardiovascular risk and vulnerability of atherosclerotic plaque through nonlipid as well as lipid-lowering mechanisms, but their anti inflammatory effects on the vascular tissue have not been fully elucidated. We investigated the effects of fluvastatin on IL-6 synthesis in human vascular smooth muscle cells (VSMCs). Addition of fluvastatin decreased IL-6 synthesis in VSMCs in a time (0-24 hours)- and dose (10(-8)-10(-5) mol/L)-dependent manner. Fluvastatin also decreased IL-6 mRNA expression in VSMCs. The effects of fluvastatin on IL-6 expression were completely reversed in the presence of mevalonate or geranylgeranyl-pyrophosphate, but not squalene. Inhibition of Rho by C3 exoenzyme or Rho kinase by Y-27632 significantly decreased IL-6 expression in VSMCs. In conclusion, fluvastatin decreases IL-6 synthesis in human VSMCs through inhibition of Rho pathway. These findings suggested that reduction of IL 6 expression by statins may partially explain their therapeutic effects in patients with coronary artery disease. PMID- 12090905 TI - High density-lipoprotein subfractions of patients using cardio-selective beta blockers. AB - Treatment of hypertension with beta-adrenergic blockers (BB) slightly increases plasma triglycerides and decreases high density lipoprotein (HDL) cholesterol levels. However, only little is known about BB-related lipid changes in patients with coronary artery disease (CAD), who usually a priori have decreased HDL cholesterol levels; and even less data exist on HDL subfraction cholesterol in these patients. We therefore quantified levels of lipids, lipoprotein lipids including HDL2 and HDL3 cholesterol, and apolipoproteins in 107 consecutive men undergoing elective coronary angiography. Of the 107 patients, 84 had angiographically established coronary atherosclerosis (>or=1 lesion with >or=50% narrowing, CAD+), and 23 had no major lesion (CAD-); 67 were taking ss1-selective BB (metoprolol or atenolol) for treatment of angina and/or hypertension and 40 were not. Patients using BB had significantly higher cholesterol levels than patients not using BB (5.99 +/- 0.93 vs. 5.63 +/- 1.07 mmol/l, mean +/- SD, p = 0.029). Their HDL cholesterol and HDL2 cholesterol levels were significantly lower (1.19 +/- 0.27 vs. 1.28 +/- 0.33 mmol/l, p = 0.048, and 0.22 +/- 0.12 vs. 0.27 +/- 0.18 mmol/l, p = 0.038, respectively). Accordingly, the total cholesterol/HDL cholesterol ratio was significantly higher in patients taking BB than in those not taking BB (5.23 +/- 1.27 vs. 4.68 +/- 1.63, p = 0.010). Considering CAD+ and CAD- patients separately, there was a trend towards lower HDL cholesterol and its subfractions with significantly lower HDL2 cholesterol in patients with BB in the CAD- group, suggesting a stronger dyslipidemic effect of BB in these patients with a priori normal or near normal baseline lipid levels. PMID- 12090906 TI - Exercise does not affect plasma concentrations of (R)- and (S)-carvedilol. AB - PURPOSE: In vitro studies have shown that beta-blockers are taken up into and released from adrenergic cells. As a consequence, plasma concentrations of beta blockers increase during exercise together with those of epinephrine and norepinephrine. However, effects of exercise on plasma concentrations of (R)- and (S)-carvedilol are unknown. METHODS: Twelve healthy males received oral single doses of 12.5 mg (R)-carvedilol, 12.5 mg (S)-carvedilol and 25 mg (R,S) carvedilol in a cross-over fashion; 11 patients with essential arterial hypertension were given 25 mg (R,S)-carvedilol. Exercise was performed 3 hours following drug intake, and blood samples were taken at rest, at the end of exercise, and after 15 min of recovery. Plasma concentrations of (R)- and (S) carvedilol were determined by HPLC. RESULTS: Plasma concentrations of (R) carvedilol were 2- to 3-fold higher than those of (S)-carvedilol (p < 0.05 in all cases). Plasma concentrations of both (R)- and (S)-carvedilol remained unaffected during exercise and recovery. CONCLUSIONS: Contrary to all other beta-blockers so far investigated, exercise had no effect on plasma concentrations of (R)- and (S) carvedilol. We conclude that neither (R)- nor (S)-carvedilol is released from adrenergic cells during exercise, a feature that clearly distinguishes carvedilol from other beta-blockers. Thus, the human organism appears to handle (R)- and (S) carvedilol differently than other beta-adrenoceptor antagonists. This finding deserves further investigation on a molecular and cellular level in order to clarify these differences between the pharmacokinetics of carvedilol and other beta-blockers. PMID- 12090907 TI - Fosinopril improves left ventricular diastolic function in young mildly hypertensive patients without hypertrophy. AB - To clarify whether fosinopril monotherapy can improve left ventricular diastolic function (LVDF) in young mildly hypertensives without hypertrophy, we studied 66 patients (pts) with diastolic blood pressure 90-100 mmHg, aged <45 years, with normal 2-dimensional echocardiography (2-D echo), and impaired DF. Impaired DF was defined as a Doppler transmitral early (E) to atrial (A) filling velocity ratio (E/A ratio) <1. Thirty-eight pts were selected for fosinopril monotherapy. Mean age was 36 years. Duration of documented hypertension was 5.4 years. Mean daily dose of fosinopril was 20 mg. Twenty-eight controls were treated with hydrochlorothiazide and hydralazine combination. Sixty-six age- and sex-matched healthy subjects served to establish normal reference values of 2-D and Doppler echo measurements. All hypertensives were treated for 30 months and re-examined 4 weeks after cessation of treatment. The fosinopril-treated group showed improvements in transmitral E (52 +/- 8 cm/s, vs. 61 +/- 9 cm/s, p < 0.01), A (56 +/- 9 cm/s, vs. 47 +/- 6 cm/s, p < 0.05), and E/A ratio (0.93 +/- 0.16, vs. 1.29 +/- 0.18, p < 0.01). Moreover, the early to atrial velocity-time integral ratio (1.31 +/- 0.10, vs. 2.24 +/- 0.10, p < 0.001) improved. The pulmonary venous flow pattern normalized after fosinopril therapy. LV mass index, relative wall thickness, LV dimension, left atrial dimension, fractional shortening, heart rate, and body mass index did not change. The hydrochlorothiazide-hydralazine combination-treated group did not show an improved diastolic function. It is concluded that long-term fosinopril monotherapy leads to an improvement of impaired LVDF in young mildly hypertensives without hypertrophy. PMID- 12090909 TI - Acute delirium induced by metoprolol. AB - OBJECTIVE: To describe a case of delirium associated with use of metoprolol and to analyse 24 such cases including 22 cases reported to Australian Adverse Drug Reaction Advisory Committee and one case previously published (S. Ahmad, Am Fam Physician, 1991;44:1142, 1144). CASE SUMMARY: An 89 year old caucasian man with an acute coronary syndrome who had no psychiatric history and no infections, brain injury, stroke, metabolic nor neoplastic disease developed delirium after two small doses of metoprolol (25 mg). The delirium disappeared within 20 hours after metoprolol was ceased, despite continuing all other medications. THE COMBINED SERIES: Of 24 patients (12 women, mean age 71.8 years), 83% were older than 60 years. The duration of therapy before onset of delirium in 14 (58%) subjects was within one week; 23 of 24 patients were receiving therapeutic amounts of the drug (25-200 mg/day). Clinical features included confusion/disorientation in all subjects, agitation in 13, aggression in 6, visual hallucinations in 7, auditory hallucinations in 1, paranoid delusions in 3, vivid dreams in 2 and language disturbances in 3 persons. Bradycardia was reported in 4 cases, hypotension in 2, fatigue/tiredness in 3, Raynaud's phenomenon in 1 and skin rash in 1 patient. DISCUSSION: The mechanism of metoprolol-induced delirium is unclear. It could be due to impairment of hepatic metabolism (especially in the ageing liver) and complex neurotransmitter-related effects on brain beta-adrenoceptors and serotonin (5-HT) receptors. CONCLUSIONS: Physicians should be aware that metoprolol, a widely used beta-blocker, may rarely cause delirium, especially in the elderly population. PMID- 12090908 TI - Pathophysiologic and therapeutic importance of tissue ACE: a consensus report. AB - Angiotensin-converting enzyme (ACE) activation and the de novo production of angiotensin II contribute to cardiovascular disease through direct pathological tissue effects, including vascular remodeling and inflammation, as well as indirect action on nitric oxide bioavailability and its consequences. The endothelium plays a pivotal role in both vascular function and structure; thus, the predominant localization of ACE to the endothelium has implications for the pathobiology of vascular disease, such as coronary artery disease. Numerous experimental studies and clinical trials support the emerging realization that tissue ACE is a vital therapeutic target, and that its inhibition may restore endothelial function or prevent endothelial dysfunction. These effects exceed those attributable to blood pressure reduction alone; hence, ACE inhibitors may exert an important part of their effects through direct tissue action. Pharmacologic studies show that while ACE inhibitors may differ according to their binding affinity for tissue ACE the clinical significance remains to be determined. PMID- 12090910 TI - Eradication of Chlamydia pneumoniae from coronary artery endothelium. PMID- 12090913 TI - Prostaglandins and mechanisms of preterm birth. AB - Increased uterine contractility at term and preterm results first from activation and then stimulation of the myometrium. Activation can be provoked by mechanical stretch of the uterus, and by an endocrine pathway resulting from increased activity of the fetal hypothalamic-pituitary-adrenal axis. In sheep fetuses, increased cortisol output during pregnancy regulates expression of prostaglandin synthase type 2 (PGHS-2) in the placenta in an oestrogen-independent manner, resulting in increased concentrations of prostaglandin E2 (PGE2) in the fetal circulation. Later increases in maternal uterine expression of PGHS-2 require increases in oestrogen and lead to increased concentrations of PGF(2alpha) in the maternal circulation. Thus, regulation of PGHS-2 at term is differentially controlled in fetal (trophoblast) and maternal (uterine epithelium) tissue. This difference may reflect expression of glucocorticoid receptor but not oestrogen receptor (ER) in placental trophoblast cells. In women, cortisol also contributes to increased prostaglandin production in fetal tissues through upregulation of PGHS-2 (amnion and chorion) and downregulation of 15-OH prostaglandin dehydrogenase (PGDH; chorion trophoblasts). The effect of cortisol on expression of PGDH in the chorion reverses a tonic stimulatory effect of progesterone, potentially through a paracrine or autocrine action. In membranes, cortisol may be derived from cortisone through activity of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) type 1, in addition to secretion from the maternal or fetal adrenal glands. In placenta, 11beta-HSD-2 oxidase activity predominates and expression of this enzyme is reduced with hypoxaemia and in placentae from pre-eclamptic pregnancies. In these circumstances, increased concentrations of maternal cortisol may cross into the fetal compartment, contributing to growth restriction and programming later life disease. PMID- 12090914 TI - Caspase-mediated apoptosis in the vertebrate ovary. AB - In the vertebrate ovary, apoptosis is the process by which excess or non-viable germ and granulosa cells are eliminated early in ontogeny (often beginning before birth), and thereafter continuously throughout reproductive life. Accordingly, an excessively high rate or abnormal triggering of such cell death (and, by implication, follicle atresia) can negatively affect fertility. Programmed cell death involves the integration of many pathways and intracellular proteins, and central among these at almost every stage are members of the caspase family. Relatively little attention has been focused upon the ovary with regards to elucidating initiator and effector members of the caspase family, and pathways by which they are activated and inactivated. The present review briefly describes vertebrate caspases and the regulation of their function in non-ovarian tissues. Subsequently, the status of caspase expression and function in orchestrating apoptotic cell death in ovarian germ and follicle somatic cells is considered. The most compelling results implicating specific caspases in ovarian function have been derived from mouse single and double knockout model systems. The final outcome of continued studies, in addition to providing information regarding understanding and management of infertility, will influence the development of strategies to treat ovarian cancers and ameliorate the adverse effects of their therapy (for example, chemotherapy). PMID- 12090915 TI - Could multiple low-affinity bonds mediate primary sperm-zona pellucida binding? AB - The current model for primary sperm binding to the zona pellucida is that a cell surface sperm protein binds with high affinity in an order-specific manner to one of the zona pellucida proteins, ZP3. However, the molecular details of primary sperm-zona pellucida binding remain elusive. A possible revised model is that multiple low-affinity bonds between spermatozoa and the zona pellucida may be sufficient for primary binding. The avidity of several low-affinity bonds can exceed that of a single high-affinity bond, which is sufficient to tether a motile spermatozoon. A mechanism involving multiple low-affinity bonds could account for some of the difficulties in elucidating primary sperm-zona pellucida interactions. PMID- 12090916 TI - Effect of maternal undernutrition on fetal testicular steroidogenesis during the CNS androgen-responsive period in male sheep fetuses. AB - The aim of this study was to determine the effects of maternal undernutrition, applied during physiologically relevant stages of development of the reproductive system, on reproductive development in male sheep fetuses. Groups of ewes (n = 11 19) were fed rations providing either 100% (high; H) or 50% (low; L) of metabolizable energy requirements for live weight maintenance during selected 'windows', bounded by days 0, 30, 50, 65 and 110 after mating. Ewes of control groups (HH (Expts 1 and 2) and HHH (Expt 3)) were fed the H ration from mating until they were killed at day 50 (Expt 1), day 65 (Expt 2) or day 110 (Expt 3) of gestation, whereas ewes of other groups were fed the L ration for the periods days 0-30 of gestation (LH and LHH), days 31-50 or days 31-65 of gestation (HL and HLH), days 65-110 of gestation (HHL), or day 0 to day 50, day 65 or day 110 of gestation (LL and LLL) when the animals were killed. At day 50 of gestation, there was no effect of nutritional treatment on mean fetal mass or fetal testicular mass, but there was increased expression of mRNA for steroidogenic acute regulatory protein (StAR) in the testes of LL animals (P < 0.05) compared with HH controls. Compared with HH animals, the mean plasma testosterone concentrations of LL fetuses tended to be higher, but this result did not reach significance. At day 65 of gestation there were no significant differences between treatments in mean fetal masses, testicular masses, mean plasma testosterone concentrations or StAR mRNA content. At day 110 of gestation, fetal masses in the LLL group were lower (P < 0.01) than those of control fetuses, although no differences in testicular size or fetal plasma testosterone concentrations were recorded. It is concluded that the effects of undernutrition on reproductive development of male sheep fetuses are dependent on the timing of the period of undernutrition. PMID- 12090917 TI - Effect of enucleation procedures and maturation conditions on the development of nuclear-transferred rabbit oocytes receiving male fibroblast cells. AB - Enucleated oocytes matured in vitro, from which chromosomes were removed by treatment with ionomycin and demecolcine, were used as recipient oocytes for nuclear transfer of fibroblast cells from a mature male rabbit. The enucleated oocytes with donor nuclei were electrically activated 2 h after fusion. The potential of nuclear-transferred oocytes matured in vitro and ovulated oocytes to develop into blastocysts was high (33-55%), except for oocytes cultured for 8.0 (19%) and 8.5 h (25%) in vitro. After transfer of nuclear-transferred oocytes to recipients, ten of 62 (16%) and one of eight (13%) recipients that received in vitro-matured and ovulated oocytes, respectively, had 19 (1%) and one (0.6%) implantation sites at the time of laparotomy on days 8-17 after transfer. Four fetuses, including two with beating hearts, were obtained on day 15 of gestation after transfer of nuclear-transferred oocytes matured in vitro. The reason for the low efficiency of fetus production was not clear. One possibility is chromosomal abnormalities of nuclear-transferred oocytes, as most (21 of 22) of the oocytes had chromosomes dispersed along the spindle fibre at the first cell cycle. This is the first report of successful production of fetuses after nuclear transfer of rabbit somatic cells. PMID- 12090918 TI - Investigation of the role of nitric oxide synthase 2 in pregnancy using mutant mice. AB - Nitric oxide (NO) has been implicated as a signalling molecule in many cellular processes. As nitric oxide synthase 2 (NOS-2) is the main isoform expressed in mouse decidua and metrial gland, mice with a targeted disruption of the gene encoding NOS-2 were used to determine the potential roles of this enzyme during pregnancy. Reproductive success and the morphology of implantation sites throughout pregnancy were compared in NOS-2 deficient (NOS-2-/-) and wild-type (WT) mice. Although there were no significant differences in the duration of gestation or birth weight, NOS-2-/- mice had significantly fewer viable embryos at mid-gestation and delivered smaller litters than did WT mice. Histological sections of uteroplacental units from WT and NOS-2-/- mice were compared to establish the mechanisms underlying the loss of fetuses. No morphological differences were observed on day 6 or day 8 of gestation, indicating that implantation and early development of implantation sites were unaffected by the absence of NOS-2. However, by mid-gestation, decidua of NOS-2-/- mice had reduced cellularity and their decidual arteries had abnormally thickened walls. These observations were quantified by morphometric measurements, which showed a significant reduction in decidual cellular area and a significant increase in the blood vessel wall:lumen ratio in NOS-2-/- mice. The increase in the thickness of the blood vessel walls was not due to abnormal cellular infiltration or to altered expression of alpha-actin in vascular smooth muscle. These results indicate that NOS-2 has a functional role in the maintenance of decidual cellular integrity and development of appropriate uterine vasculature, and may play a supportive role in promoting embryo survival. PMID- 12090919 TI - Partial cloning and polymorphism of the melatonin1a (Mel1a) receptor gene in two breeds of goat with different reproductive seasonality. AB - The melatonin1a (Mel1a) receptor gene was cloned in two breeds of goat, one with marked seasonal ovarian activity (Alpine breed) and the other with low seasonal variations in ovulatory activity (Creole breed), to determine whether reproductive seasonality is related to the structure of the Mel1a gene. The main part of exon II was amplified by PCR using sheep sense and antisense primers in 17 Alpine and 13 Creole goats, and cDNAs were subcloned and sequenced in both directions. The results indicate the presence of an identical sequence in 12 of the 30 animals, that is, six Alpine and six Creole goats. The greatest similarity in the 784 nucleotides of exon II (primers excluded) that was obtained was found with ovine Mel1a receptor sequence (98.4%) and the differences consisted of 12 nucleotide and four amino acid changes. The presence of seven mutations compared with the previous reference sequence was observed and their combinations indicated the presence of at least five other alleles; one mutation resulted in a change in one amino acid in three Alpine goats. No difference in allelic distribution was observed between the two breeds. The results indicate that no relationship could be established between the Mel1a receptor gene structure and the expression of seasonality of reproduction in goats. PMID- 12090920 TI - Alleviation of the two-cell block of ICR mouse embryos by polyaminocarboxylate metal chelators. AB - The present study was undertaken to examine the effects of various transition metal ion chelators, both polyaminocarboxylates (including nitrilotriacetate (NTA), ethylenediaminediacetate (EDDA), ethyleneglycolbistetraacetate (EGTA), ethylenediaminetetraacetate (EDTA) and diethylenetriaminepentaacetate (DTPA)) and non-polyaminocarboxylates (dipicolinic acid and deferoxamine), on the development in vitro of one-cell ICR strain mouse embryos to the four-cell and blastocyst stages. The order of stability constants of polyaminocarboxylates for transition metal ions such as zinc, copper and iron is as follows: NTA < or = EDDA < EGTA < EDTA < DTPA. Addition of 10 or 100 micromol polyaminocarboxylates x l(-1) to the medium significantly enhanced the development of most one-cell embryos (66-88%) beyond the two-cell stage compared with that (< 25%) in medium without polyaminocarboxylates. Although EDDA, EDTA and DTPA at 10 micromol x l(-1) induced the development of most one-cell embryos to the four-cell stage and beyond, a higher concentration (100 micromol x l(-1)) of NTA and EGTA was required to obtain a similar result. Therefore, the ability of polyaminocarboxylates to overcome the two-cell block is not correlated with their potency to chelate transition metal ions. In contrast, the non polyaminocarboxylates dipicolinic acid and deferoxamine, at 10 and 100 micromol x l(-1), did not have the same effect. Taken together, the results indicate that the ability of polyaminocarboxylates to overcome the two-cell block in embryo development is due to some common feature or features other than the ability to chelate transition metal ions. PMID- 12090921 TI - Effects of oestrogen treatment on testicular descent, inguinal closure and prostatic development in a male marsupial, Macropus eugenii. AB - This study reports the effect of oestrogen treatment on the development of the genital ducts, prostate gland, testicular descent and inguinal canal closure in male tammar wallaby young treated with oestrogen over four time spans during the first 25 days of pouch life (days 0-10, 10-15, 15-25 and 0-25) and sampled at day 50. In control males, the Mullerian ducts had regressed and the Wolffian ducts had developed into the vas deferens and epididymis. The prostate gland had formed epithelial buds extending from the ventral, lateral and posterior walls of the urethra. The testes were in the neck of the scrotum and the gubernaculum and processus vaginalis were present at the base of the scrotum. In most males treated with oestradiol from day 0 to day 25, the testes had failed to descend by day 50. The gubernaculae were long and thin. The retained Mullerian ducts formed a lateral vaginal expansion like that of normal day 50 females. The Wolffian ducts of the males treated on days 0-25 were regressed, but were present in males in the other three treatment groups. The prostate glands were hyperplastic and epithelial budding was highly invasive. Some treated males from the day 10-25 and 0-25 groups had inguinal hernias. These results demonstrate that oestrogen treatment has profound effects on the development of the internal genitalia of a male marsupial, preventing inguinal closure and interfering with testicular descent. Therefore, the tammar wallaby may provide a useful experimental model animal in which to investigate the hormonal control of testicular migration and closure of the inguinal canal. PMID- 12090922 TI - Isolation and purification of type A spermatogonia from the bovine testis. AB - The aim of this study was to isolate and purify bovine type A spermatogonia. Testes from 5-7-month-old calves were used to isolate germ cells using a two-step enzymatic digestion. During the isolation and purification steps, the viability of cells was determined using live/dead staining. The identity of type A spermatogonia during isolation and purification was determined under a light microscope equipped with a Nomarski lens. Isolated cells were characterized further by using specific markers for type A spermatogonia, including Dolichos biflorus agglutinin (DBA) and c-kit. The cell suspension was transplanted into immunodeficient recipient mouse testes and the colonization was assessed 1-3 months after transplantation, to assess the stem cell population among the isolated cells. After isolation, a cell suspension was obtained containing about 25% type A spermatogonia, which was enriched further by differential plating and separation on a discontinuous Percoll gradient. Finally, fractions containing 65 87% pure type A spermatogonia were obtained. Large and small type A spermatogonia with different numbers and sizes of nucleoli were found. DBA stained both large and small type A spermatogonia and its application in fluorescence-activated cell sorting (FACS) resulted in comparable percentages of type A spermatogonia to those determined by morphological examination under a light microscope equipped with a Nomarski lens. Nearly all of the large type A spermatogonia showed strong c-kit immunoreactivity, indicating that these cells had undergone at least an initial differentiation step. In contrast, approximately half of the small type A spermatogonia were negative for c-kit, indicating the presence of the spermatogonial stem cells in this population. At 3 months after transplantation, groups of bovine type A spermatogonia were found in most tubule cross-sections of the recipient mouse testes, showing the presence of spermatogonial stem cells among the isolated cells. PMID- 12090923 TI - A quantitative (stereological) study of the effects of experimental unilateral cryptorchidism and subsequent orchiopexy on spermatogenesis in adult rabbit testis. AB - The aim of this study was to examine the controversial effects of experimental unilateral cryptorchidism and subsequent orchiopexy on the number of germ cells and other morphometric characteristics of testicular and epididymal structures in adult rabbits. Unilateral cryptorchidism was induced in 11 mature male New Zealand white rabbits by returning one testis, together with the ipsilateral epididymis, to the abdominal cavity via a surgical procedure. After 3 months, testes and epididymides were removed from six animals (and from six age-matched control animals that did not undergo the surgery). Orchiopexy was performed on the five remaining animals and the testes and epididymides of these animals (and an additional six age-matched control animals) were removed 7 weeks later. A contemporary, unbiased and efficient stereological tool, the optical disector, was used to estimate the number of nuclei in the testis and epididymis using methacrylate-embedded sections of 25 micron in thickness. Cryptorchidism resulted in severe testicular atrophy and spermatogenic arrest: type A spermatogonia and Sertoli cells only were seen in the seminiferous epithelium, and the number of type A spermatogonia per testis was reduced by 84%. After orchiopexy, the testis remained atrophied and the number of type A spermatogonia returned to the near normal range in four of five animals, but spermatogenesis was recovered only partially at the stage of early primary spermatocytes (one animal), late primary spermatocytes (two animals) or spermatids (one animal). In conclusion, cryptorchidism caused severe spermatogenic arrest that was potentially recoverable (in view of the restoration of the number of type A spermatogonia), but orchiopexy failed to induce full recovery of spermatogenesis. PMID- 12090924 TI - Induction of sperm maturation in vitro in epididymal cell cultures of the tammar wallaby (Macropus eugenii): disruption of motility initiation and sperm morphogenesis by inhibition of actin polymerization. AB - A sperm-epididymal cell co-culture was shown to be capable of inducing the in vitro maturation of spermatozoa from a marsupial species, the tammar wallaby (Macropus eugenii). This system was able to maintain wallaby epididymal epithelial cells in vitro for more than 2 months. The system also enabled immature wallaby spermatozoa to differentiate from a T-shaped to a streamlined form, accompanied by the development of progressive motility after co-culture with epididymal cell monolayers that had been cultured for 7 days. The addition of inhibitors of actin polymerization (latrunculin A or B) to the co-culture system showed that wallaby sperm maturation was impaired by the interruption of actin organization within the immature spermatozoa. These results indicate that actin filaments play a significant role in sperm transformation during post testicular maturation in marsupials. These observations also indicate that the marsupial co-culture system has the potential to greatly increase understanding of sperm-epididymal cell interactions and the mechanism of sperm maturation in these species. PMID- 12090926 TI - Effect of local heating of rat testes after suppression of spermatogenesis by pretreatment with a GnRH agonist and an anti-androgen. AB - The effects of local heating of rat testes, in which spermatogenesis had been suppressed with injections of a GnRH agonist and an anti-androgen, were examined. Although the detrimental effects of heating were not as marked as those found in the testes of non-injected rats, the testes in which spermatogenesis was suppressed also showed a significant reduction in mass, the number of spermatozoa, tubular diameter and the percentage of normal tubular cross-sections at day 35 after heating. The results indicate that heating has an effect on cells in the testis other than those shown to be most susceptible to heat, namely pachytene spermatocytes and early spermatids, which were absent or markedly reduced in number when spermatogenesis was suppressed. The long-term effects of heating on the above parameters, as reported in a previous study, were also confirmed. However, in testes in which spermatogenesis was suppressed at the time of heating, there appeared to be no or a reduced long-term impairment of spermatogenesis, as determined by testis mass, the percentage of qualitatively normal tubules and epididymal sperm counts. PMID- 12090925 TI - Effects of dietary polyunsaturated fatty acids on ovarian and uterine function in lactating dairy cows. AB - Alteration of the polyunsaturated fatty acid (PUFA) composition of milk by dietary supplementation of cows may be beneficial to human health. However, dietary PUFAs may influence synthesis of both prostaglandins and steroid hormones. This study examined the effects of dietary PUFAs on reproductive parameters in lactating cows. Cows were fed an isoenergetic control ration (n = 8) or a diet supplemented with LinPreme (n = 7) or SoyPreme (n = 8). These proprietary feeds are derived from linseed or soybeans and contain high concentrations of linolenic acid (LNA, n-3) or linoleic acid (LA, n-6) protected PUFA, respectively. Both PUFA-supplemented diets reduced plasma progesterone, particularly in the early luteal phase, and increased the number of medium-sized (5-10 mm in diameter) follicles. The diameter of the first dominant follicle, insulin-like growth factor I (IGF-I) concentrations at oestrus and cholesterol concentrations were all higher in cows fed a diet supplemented with LA (n-6) than in cows that did not receive this supplement. In cows fed a diet supplemented with LNA (n-3), there was an increase in oestradiol during the follicular phase. Diet had no effect on non-esterified fatty acid or insulin concentrations, or on the duration of the oestrous cycle. The plasma concentration of 13,14,dihydro-15 keto PGF(2alpha) after administration of 50 iu oxytocin was unaffected by diet on day 15 and day 16 of the oestrous cycle, but showed a greater response on day 17 in the LA (n-6) supplemented group. Therefore, the PUFA content of the diet can influence both ovarian and uterine function in cows. However, further studies using larger numbers of cows are required to test whether fertility is also affected by such diets. PMID- 12090927 TI - Effects of high concentrations of hyaluronan in culture medium on development and survival rates of fresh and frozen-thawed bovine embryos produced in vitro. AB - Hyaluronic acid (HA) is the main glycosaminoglycan present in follicular, oviductal and uterine fluids. The main functions of HA include dynamic processes that are mediated through interaction with extracellular matrix components, regulation of gene expression, cell proliferation and cell differentiation. HA increases the viscosity of solutions and also has several physiological functions, including regulation of water distribution and water-binding capacity. The addition of 6 mg HA ml(-1) to synthetic oviduct fluid (SOF; SOF-HA) culture medium on day 5 (IVF = day 0) significantly (P < 0.001) increased the viscosity of the medium in comparison with SOF culture medium containing BSA (SOF-BSA). On day 8, rate of blastocyst development in SOF-HA culture medium was significantly (P < 0.05) higher than in SOF-BSA culture medium (38.2 versus 29.3%). The number of trophectoderm cells and the total number of cells of expanded blastocysts cultured in the presence of HA were significantly (P < 0.01) higher in comparison with expanded blastocysts cultured in the presence of BSA (88.9 +/- 7.3 versus 67.6 +/- 3.0 and 130.1 +/- 10.9 versus 104.8 +/- 2.5, respectively). After freezing and thawing, the percentage of day 8 embryos that re-expanded and hatched when cultured with SOF-HA was greater than that of embryos cultured with SOF-BSA (11.3 and 10.5% versus 75.5 and 36.8%, respectively). After thawing, the ATP contents of in vivo-derived, SOF-HA and SOF-BSA expanded blastocysts were similar. The embryos cultured with HA showed less ultrastructural deviation and de-differentiation after freezing and thawing than the embryos cultured with BSA. This study demonstrates that HA improves the developmental capacity of bovine embryos under in vitro conditions and is warranted as a supplement for in vitro production of bovine embryos, particularly if they are to be cryopreserved. PMID- 12090928 TI - Development, amino acid utilization and cell allocation in bovine embryos after in vitro production in contrasting culture systems. AB - The effects of protein-supplemented and protein-free media on amino acid uptake, protein synthesis and cell differentiation in bovine blastocysts were investigated. Four formulations of synthetic oviduct fluid were used. Each formulation was identified by the principal supplement: bovine serum albumin (0.4%, w/v); polyvinyl alcohol (0.3%, w/v); or either of two steer sera (10%, v/v). After zygote culture, blastocyst yields (day 7.5) were lowest in protein free medium and highest in albumin-supplemented medium. Subsequent 12 h incubation in the presence of both essential and non-essential amino acids was used for the measurement of amino acid flux. All blastocysts released alanine but consumed aspartate (P < 0.001) and the extent was influenced by prior culture conditions. Aspartate uptake was lower in blastocysts produced in protein-free conditions (P < 0.05) than in blastocysts produced in albumin-supplemented conditions. Consumption indices for 16 other amino acids were not influenced by blastocyst source. Cell counts and hatching incidences were highest for albumin supplemented blastocysts, but were similar among blastocysts from the protein free and serum-dependent treatments. Crucially, the use of protein-free medium for zygote culture did not compromise resultant blastocysts in terms of either de novo protein synthesis ([3H]phenylalanine incorporation) or trophectoderm function (phenotype based on interferon-tau detection). Thus, although blastocyst yields were compromised after zygote culture in a protein-free (vis-a-vis albumin supplemented) medium, amino acid flux was qualitatively conserved, and only quantitatively modified in the case of alanine and aspartate. Moreover, vital properties of blastocysts that were produced, including de novo protein synthesis and trophectodermal cell function, apparently were not adversely affected by protein deprivation. PMID- 12090930 TI - Evaluation of progesterone receptor expression in eosinophils using real-time quantitative PCR. AB - Progesterone has been shown in many instances to have immune-suppressant activities. Most of these activities have been investigated in the light of general immune suppression or with a focus on lymphocytes. However, many clinical and in vitro studies have shown that progesterone also has a suppressive effect on eosinophilia. This effect so far has not been thoroughly investigated. The purpose of this study was to evaluate whether the effect is mediated via the classical progesterone receptor (PR). We developed a new real-time quantitative PCR (RQ-PCR) for the analysis and quantification of expression of the classical PR. The test was first validated both on breast cancer cell lines and on breast cancer biopsies. Subsequently, when using eosinophils isolated from peripheral blood of healthy volunteers, we could not find evidence for the expression of PR. These data suggest that the effects of progesterone on eosinophils are not mediated by the classical PR. PMID- 12090931 TI - Evidence for a role of hyaluronan in the spacing of fibrils within collagen bundles in rabbit synovium. AB - Synovial hydraulic resistance is vital for the retention of intra-articular fluid, and originates within the matrix of biopolymers in the intercellular gaps. Specific digestion of hyaluronan resulted in a increase in synovial hydraulic permeability from 0.478+/-0.24 microl min(-1) cm H(2)O(-1) in control tissue to 4.561+/-0.40 microl min(-1) cm H(2)O(-1) (mean+/-S.D., n=6 rabbits, P<0.001 t test). To investigate whether hyaluronidase also altered the interstitial ultrastructure, morphometry of hyaluronidase treated synovium was carried out. The most striking novel finding was that hyaluronidase treatment reduced extrafibrillar volume fraction within the synovial collagen bundles from 50.5+/ 11.1% to 36.8+/-15.5% (mean+/-S.D., n=6 rabbits, P<0.001, two-way anova). This was accompanied by a reduction in interfibrillar centre to centre spacing from 101+/-11 (control) to 84+/-6 nm (mean+/-S.D.; n=6 rabbits, P<0.001) in enzyme treated bundles. Individual fibrils showed a small but highly significant reduction in cross-sectional diameter from 76.9+/-6.3 to 72.5+/-6.3 nm (mean+/ S.E.; P<0.001) after hyaluronidase treatment. The findings indicate that hyaluronan chains have a major organisational role within the collagen bundle itself. The trans-synovial pathway comprises bundles and substantial areas of intervening, bundle-free matrix, and it is possible that bundle collapse contributes to a rise in overall permeability by increasing the inter-bundle space. PMID- 12090932 TI - Application of a normalised plot to the study of uni-uni enzyme-inhibitor systems. AB - Normalisation of kinetic data is a useful tool in the study of complex enzyme systems. In the present paper, we have applied the premises of the normalised plot to the description of uni-uni enzyme inhibition. Guidelines to the design of the experiments and to data managing using the freeware program SIMFIT (http:??www.simfit.man.ac.uk) are offered. The treatment has a lessened demand in experimental data while ensuring biological consistence of the results. Moreover, the results are obtained without resorting to secondary plots, and the election between rival mechanisms is statistically granted. Hyperbolic mixed-type inhibition is studied as a general model for enzyme-inhibitor/activator interaction, and equations describing classical cases of linear inhibition are also considered. PMID- 12090933 TI - IMP-1 metallo-beta-lactamase: effect of chelators and assessment of metal requirement by electrospray mass spectrometry. AB - Metallo-beta-lactamases have attracted considerable attention due to their role in microbial resistance to beta-lactam antibiotics. IMP-1, the binuclear Zn dependent beta-lactamase produced by Pseudomonas aeruginosa and other microorganisms, is of particular interest in view of its increasing prevalence. An examination of the susceptibility of IMP-1 to inactivation by six different divalent metal ion chelators has revealed that all except Zincon cause inhibition by forming a complex with the holoenzyme. Exposure of the enzyme to dipicolinic acid (DPA), the most potent inhibitor, results in the production of the mononuclear Zn form of the protein as determined by electrospray ionization mass spectrometry (ESI-MS) under nondenaturing conditions. This mononuclear Zn species was found to be catalytically competent. Studies with the chromophoric chelator 4 (2-pyridylazo)resorcinol (PAR) show that the two zinc centers in IMP-1 differ in their accessibility, a feature that could be overcome in the presence of guanidine hydrochloride (GdnHCl, 1.5 M). PMID- 12090934 TI - Flavones from Scutellaria baicalensis Georgi attenuate apoptosis and protein oxidation in neuronal cell lines. AB - The oxidative modification of proteins plays a major role in a number of human diseases including Alzheimer's disease (AD). Flavones in extracts of Scutellaria baicalensis (SbE) have been reported to have exceptional antioxidant properties. We examined the effects of SbE on neuronal cells exposed to oxidative stress. Neuronal HT-22 cells were exposed to low levels of H(2)O(2) generated from glucose oxidase (GO) under conditions that caused cell death in 24 h. The mechanism of cell death was shown to occur via apoptosis. Flavone extracts (50 microg/ml) protected cells and increased viability to 85+/-5% (P<0.001). The flavones also increased the content of Bcl-2 in the cell, resulted in its phosphorylation, and in contrast decreased the Bax levels. Furthermore, the oxidative-stress-induced protein carbonyl formation was reduced nearly two-fold when cells were pretreated with the flavone extract. Two-dimensional electrophoresis (2-DE) showed that less than 15% of the total visible proteins were oxidized and that the oxidation was specific for certain oxidation-sensitive proteins. These data support the idea that flavones in SbE can attenuate oxidant stress and protect cells from lethal oxidant damage. PMID- 12090935 TI - Mechanisms underlying the cytotoxic effects of Tachpyr--a novel metal chelator. AB - Tachpyr (N,N'N"-tris(2-pyridylmethyl)-cis,cis-1,3,5-triaminocyclohexane), a novel metal chelator, was previously shown to deplete intracellular iron and exert a cytotoxic effect on cultured bladder cancer cells. Tachpyr binds Fe(II) and readily reduces Fe(III). The iron(II)-Tachpyr chelate undergoes intramolecular oxidative dehydrogenation resulting in mono- and diimino Fe(II) complexes. The present study investigates the redox-activity of the Tachpyr-iron complex to better define the mechanism of Tachpyr's cytotoxicity. Tachpyr's mechanism of cytotoxicity was studied using cell-free solutions, isolated DNA, and cultured mammalian cells by employing UV-VIS spectrophotometry, oximetry, spin-trapping technique, and electron paramagnetic resonance (EPR) spectrometry. The results show that: (1) Tachpyr by itself after 24 h of incubation had a cytotoxic effect on cultured cells; (2) fully oxidized Tachpyr had no cytotoxic effects on cultured cells even after 24 h of incubation; (3) Tachpyr protected isolated DNA against H(2)O(2)-induced damage, but not against HX/XO-induced damage; and (4) Tachpyr-Fe(II) chelate slows down but does not block oxidation of Fe(II), allows O*(-)(2)-induced or Tachpyr-induced reduction of Fe(III), and consequently promotes production of *OH through the Haber-Weiss reaction cycle. The results indicate that Tachpyr can protect cells against short-term, metal-mediated damage. However, upon prolonged incubation, Tachpyr exerts cytotoxic effects. Therefore, in addition to iron depletion, low-level oxidative stress, which in part occurs because of redox cycling of the coordinated iron ion, may contribute to the cytotoxic effects of Tachpyr. PMID- 12090936 TI - Chondroitin sulfate B exerts its inhibitory effect on secondary lymphoid tissue chemokine (SLC) by binding to the C-terminus of SLC. AB - We previously reported that certain glycosaminoglycans (GAGs) bind secondary lymphoid tissue chemokine (SLC, CCL21) and that the SLC-binding GAGs, including chondroitin sulfate B (CS B), negatively modulate the function of SLC, although the mechanism remains unknown [J. Biol. Chem. 276 (2001) 5228]. To gain insight into the mechanism of inhibition, we used a C-terminally truncated SLC (SLC-T) that lacked clusters of basic amino acid residues that have been implicated in GAG binding. While SLC-T failed to bind any GAGs, it induced prominent intracellular Ca(2+) mobilization in CC chemokine receptor (CCR) 7-expressing cells, as did wild-type SLC. However, the SLC-T-induced Ca(2+) influx was not inhibited by CS B, unlike the SLC-induced Ca(2+) influx. These results demonstrate the requirement of the C-terminus of SLC for the inhibition of chemokine responses by CS B; that is, CS B exerts its inhibitory effect by binding to the C-terminus of SLC, thus defining the mode of action of CS B on certain chemokines. PMID- 12090938 TI - Effect of ionic strength on the binding of ascorbate to albumin. AB - A fluorophore-nitroxide free radical dual-functional probe (FN) was utilized to study the kinetics of ascorbate (AH(-)) binding to Bovine Serum Albumin (BSA). Since the free radical fragment in the FN probe intramolecularly quenches fluorescence, ascorbate reduction of the nitroxide function is accompanied by a concomitant fluorescence intensity increase from the fluorophore. Thus, both fluorescence and the EPR techniques could be utilized to measure the reaction rate. In the presence of BSA protein, the observed rate of the overall process is the sum of that from at least two reactions: the reaction between free ascorbate and free probe, and the reaction between bound ascorbate and bound probe. Our findings show that the observed rate is strongly dependent on the ionic strength of the medium. A corollary of this observation is the indication of a purely electrostatic interaction between ascorbate and the BSA protein. This conclusion was further corroborated by 1H NMR measurement of the transverse relaxation time, T(2), of ascorbate protons in BSA solutions. Ascorbate ion was released from the ascorbate/BSA ensemble in the presence of increasing concentrations of NaCl. Binding constants of AH(-) to BSA were calculated at different ionic strengths at pH 7.4. Furthermore, an increase in ionic strength did not affect the ability of albumin to protect ascorbate against autoxidation. This suggests that the protein's protective antioxidant effect may be attributed to BSA binding of trace quantities of transition-metal cations (rather than ascorbate binding to BSA). This conclusion is supported by ascorbate UV-absorption measurements in the presence of albumin and Cu(2+) ions as a function of ionic strength. PMID- 12090939 TI - Simple quantification of Cu,Zn-superoxide dismutase activity after separation by non-denaturing isoelectric focusing. AB - The Cu,Zn-superoxide dismutase (SOD) activity in bovine retina cytosol was separated from retinal pigment using short-length non-denaturing isoelectric focusing (IEF) (15-mm long x 1.3-mm i.d. column) and detected using non denaturing two-dimensional electrophoresis (2-DE). After the SOD and pigment in the retina cytosol are separated, SOD activity can be quantified by water-soluble tetrazolium salt. We also found that SOD separated by this IEF retained its native function. PMID- 12090941 TI - Serial and simultaneous feature positive discriminations in a human Skinner box. AB - In the present experiment participants were presented either a serial (F-->T+/T-) or a simultaneous (FT+/T-) feature positive discrimination using a Skinner box for humans. After the participants mastered the discrimination, the associative properties of F were examined using a transfer test and a counterconditioning manipulation. F affected responding to a transfer target less in the Serial than in the Simultaneous condition. However, counterconditioning of F did not affect initial discrimination performance in either condition. These results were discussed in the framework of occasion setting, elemental learning, configural learning, and a neural network model. PMID- 12090937 TI - Evidence for binding of the ectodomain of amyloid precursor protein 695 and activated high molecular weight kininogen. AB - To identify ligands that bind to the N-terminal portion of human amyloid precursor protein (APP), we sought binding partners for a fragment of the ectodomain of human APP695 (sAPP(695)T). The probe bound to fragments of high molecular weight kininogen (HK) in rat cortical membrane preparations in vitro. Laser confocal microscopy indicated that APP and HK colocalize near cerebral blood vessels, in the neuropil, and in many neurons of rat brain. sAPP(695)T bound to human activated kininogen (HKa) (K(d)=0.3+/-0.1 nM), but not to inactivated or low molecular weight kininogen. Binding was specific for the light chain sequence of HKa. Biotinylated human HKa also bound to sAPP(695) (K(d)=0.3+/ 0.5 nM). sAPP(695) and HKa form tight complexes in solution that can be coimmunoprecipitated. These results support the hypothesis that forms of APP and kininogen can interact in brain tissue. Considering the implications of APP in neurite outgrowth, the APP-HKa interaction could modulate neurogenesis. PMID- 12090942 TI - Autoshaping in the rat: conditioned licking response to a stimulus that signals sucrose reinforcement. AB - The present experiments were designed to determine if repeated presentations of an empty sipper tube (the conditioned stimulus or CS) with the response independent delivery of a sucrose solution (the unconditioned stimulus or US) from a second spout results in the development of Pavlovian conditioned responding. In Experiment 1, rats in the experimental condition received paired CS-US presentations whereas subjects in the control condition were exposed to random presentations of CS and US. In Experiment 2, an additional control condition (CS alone) was included and, to encourage generalized responding between the US and CS, the CS tube was filled with water for all groups. The results of both experiments indicate that the CS-directed responding in the paired CS-US condition was Pavlovian in nature. Thus, the present procedure serves as an autoshaping task in which conditioned licking is generated. PMID- 12090943 TI - Self-control and impulsiveness with asynchronous presentation of reinforcement schedules. AB - IN DISCRETE TRIALS, PIGEONS WERE PRESENTED WITH TWO ALTERNATIVES: to wait for a larger reinforcer, or to respond and obtain a smaller reinforcer immediately. The choice of the former was defined as self-control, and the choice of the latter as impulsiveness. The stimulus that set the opportunity for an impulsive choice was presented after a set interval from the onset of the stimulus that signaled the waiting period. That interval increased or decreased from session to session so that the opportunity for an impulsive choice became available either more removed from or closer in time to the presentation of the larger reinforcer. In three separate conditions, the larger reinforcer was delivered according to either a fixed interval (FI) schedule, a fixed time (FT) schedule, or a differential reinforcement of other behavior (DRO) schedule. The results showed that impulsive choices increased as the opportunity for such a choice was more distant in time from presentation of the larger reinforcer. Although the schedule of the larger reinforcer affected the rate of response in the waiting period, the responses themselves had no effect on choice unless the responses postponed presentation of the larger reinforcer. PMID- 12090944 TI - The effect of study-test modalities on the remembrance of subjective duration from long-term memory. AB - It was examined whether stimulus modality (auditory vs. visual) affects the retrieval of subjective duration from memory. In two experiments the temporal generalization paradigm was used. Participants had to decide whether the previously learned standard duration (400 ms) occurred in the context of comparison stimuli. Two major results were found. (1) Discrimination was more accurate if the training and testing stimuli were of the same modality than if they were of opposite modalities. (2) If both modality of learning and modality of testing were different, subjects systematically underestimated the test durations, i.e. temporal generalization gradients (the proportion of identifications of a stimulus as the standard, plotted against stimulus duration) shifted to the right. The observed shift is interpreted as a result of a delayed timing process. PMID- 12090945 TI - Retroactive interference after discrimination reversal decreases following temporal and physical context changes in human subjects. AB - One experiment was conducted to test the additive effects of physical context changes and the passage of time on a retroactive interference task in human subjects. Participants learned a discrimination in a symbolic matching to sample situation within a specific context. The discrimination was subsequently reversed. The context in which the reversal occurred was combined factorially with the passage of time before the test. All testing was conducted in the context in which the original discrimination was acquired. Participants had received the discrimination reversal in either a context different from that in which the original discrimination was acquired, or in the same context. Half of each of the groups mentioned above received testing immediately after reversal training and the other half received testing 48 h later. Both manipulations, changing the context after the reversal and the passage of time following the reversal, led to a recovery of the original discrimination performance. Participants that received both a context change and retention interval showed the largest recovery. PMID- 12090946 TI - Post-traumatic orbital floor reconstruction with nasoseptal cartilage in children. AB - OBJECTIVE: Repair of orbital floor fractures may require the placement of a graft or implant. Both autogenous and alloplastic materials have been used for this purpose. This article reports the use of nasal septal cartilage for the repair of orbital floor defect secondary to blunt facial trauma in children. METHODS: Three children with disruption of the orbital floor after facial trauma were included in this prospective review. All children underwent open reduction with rigid fixation of the facial fractures and reconstruction of the orbital floor with nasoseptal cartilage. RESULTS: All of the cases were successfully treated by restoration of the orbital floor continuity. On follow-up clinical examination, one patient had persistent mild enophthalmus. CONCLUSIONS: Nasal septal cartilage is a readily accessible autogenous material with minimal donor site morbidity, and should be considered when an autogenous orbital implant is needed for the repair of a traumatic orbital floor defect. PMID- 12090947 TI - Effects of parental style of interaction on language development in very young severe and profound deaf children. AB - AIMS: (1) To study the influence of different styles of parent-child interaction in the language development of very young deaf children. (2) To find out if there are differences in parent-child interaction between two groups of very young deaf children following an Aural/Oral or a Bilingual approach to education. METHODS: Subjects were selected from all deaf children in the County of Avon who were under 3 years of age at the time of first assessment, had severe or profound, bilateral, sensorineural hearing loss and no associated medical problems. There were 16 children and families at the start of the project but only 13 completed all the assessments. The Bristol Language Developmental Scales (BLADES) was used to assess both sign and spoken language development. Interaction was studied through analysis of contingency and book-reading applied to selected periods of 3 min from four videorecorded sessions, taken at 3 months intervals for a period of 1 year. RESULTS: From the 13 children studied, only seven presented with some degree of expressive language measurable by the BLADES. Analysis of contingency showed that parents present with higher percentage of both Direct Related Acts and ON then their children Acts (On Acts: where both individuals are involved in the same task). Regarding bookreading, it was observed that parents often attend to child initiatives and acknowledge most of them but they make little effort to expand or use the child's message as topic for further conversation. In the reduced sample of seven children with expressive language, those with better language development had parents with: (a) higher percentage of DR acts; (b) higher percentage of ON acts; (c) higher percentage of appropriate responses to child communicative initiatives. CONCLUSIONS: In this small group language development seems to be facilitated by encouraging child participation and using a more contingent and child centred interaction. No significant differences were found between oral and bilingual families in terms of quality of interaction. PMID- 12090949 TI - Distortion evoked otoacoustic emission using GSI 70 analyzer for neonatal screening. AB - OBJECTIVE: Following the recommendation of the United State National Institute of Health Consensus Conference in 1993, otoacoustic emissions (OAE) are now used internationally for hearing screening. The GSI 70 OAE screener provides the means for carrying out OAE recordings within a short period of time and includes an automatic evaluation of results. The aim of this study was to determine the reliability of recordings in comparison with available standards in brainstem audiometry. METHOD: OAE recorded in 29 hearing-impaired suspected infants and young children (aged 1 months-7 years old) in order to compare the result of the GSI 70 screener with the result of ABR. This study was conducted in the outpatient clinic of the Tokyo University Hospital. RESULT: Our study showed that the GSI 70 screener has higher false negative rate compared with ABR results (P<0.01). Consequently, an OAE measuring method, is also provides high sensitivity and easy to use. However, there was no trend toward increased refer rates with increased age. CONCLUSION: Our findings show that the GSI 70 screener can meet the demands of systematic hearing screening in infants and young children, although there is a tendency to miss cochlear impaired cases. PMID- 12090948 TI - Anomaly of the eustachian tube and its associated structures in patients with multiple congenital malformation: a histopathological and morphometric study. AB - Two temporal bone-eustachian tube (ET) specimens; one from a 1-day-old female newborn with Townes Syndrome (Case 1) and the other from a 15-year-old female with oral-facial-digital syndrome (Case 2) were studied histopathologically and morphometrically. Both specimens had anomalies of the ET and its associated structures as compared with eight age-matched control cases without anomaly (six cases for Case 1 and two cases for Case 2, respectively). Case 1 had a weak attachment of the tensor veli palatini muscle (TVPM) to a poorly developed lateral lamina (LL) of the ET cartilage, a large voluminous medial lamina (ML) of the ET cartilage, a small voluminous ET lumen with a few ridges of mucosal folds, poorly developed ET glands and a poorly developed levator veli palatini muscle (LVPM). Case 2 had a short and longitudinally elongated ET lumen that was insufficiently covered with a poorly developed ML and LL and an aberrant course of the LVPM. Both cases were accompanied by a mild inner ear anomaly (slightly shortened cochlea). We discuss the implications of the observed anomalies with regard to functional and clinical issues. In particular, we speculate that these ET anomalies may closely be related to potential ET dysfunction with high susceptibility to otitis media. PMID- 12090950 TI - Esophageal foreign bodies: a Jordanian experience. AB - OBJECTIVES: To study the pattern of esophageal foreign bodies (FB) in Jordan and to compare it with other countries. METHODS: This is a retrospective study conducted on all patients (527) who were admitted with esophageal FB during a period of 9 years from January 1992 to January 2000. Two major referral hospitals were involved: the Princess Basma and Al-Bashir hospitals. A data sheet was constructed in which we included: name, age, gender, presenting symptoms, type and site of FB, technique of removal, and complications if present. RESULTS: Of the 527 patients 53% were male and 47% were female.77% were children under the age of 10 years. In children coins were by far the most common FB to be found in the esophagus (68%), while bones were the principal FB to be found in adults (8%). In 89%, the FB was found at the level of cricopharyngeal muscle. Drooling of saliva (72%), dysphagia (71%), and vomiting (24%) were the most commonly presenting symptoms. Both rigid esophagoscopy and Magill forceps techniques were used to remove the FB from the esophagus. The complication rate was 2% which included: esophageal perforation and mediastinitis, esophageal stenosis and esophageal erosions. CONCLUSION: The pattern of esophageal FB in Jordan is not different from other countries. PMID- 12090951 TI - Bronchial foreign body presenting as an accidental radiological finding. AB - The aspiration of a foreign body in an airway is usually associated with respiratory distress, wheeze and persistent cough. The highest incidence of foreign body inhalation occurs between the age of 1 and 3 years [Ann. Otol. Rhinol. Laryngol. 89 (1980) 434: Med. J. Aust. 2 (1983) 322]. Asymptomatic and long standing foreign bodies may lead to complications such as recurrent pneumonia, bronchiectasis, atelectasis and even death. We present here a case of a metallic bronchial foreign body, which was discovered only as an accidental radiological finding on a chest X-ray, which was done for a mild lower respiratory tract infection, presumably 4 months after such an aspiration. PMID- 12090952 TI - Endoscopic removal of esthesioneuroblastoma. AB - Esthesioneuroblastoma is a rare tumor of neural crest origin that arises in the nasal cavity. There is still no consensus on the optimal treatment for this neoplasm, and the literature contains very few accounts of endoscopic excision in these cases. We described a case report of 12-year-old girl with esthesioneuroblastoma that was confined to the nasal cavity and paranasal sinuses, with no orbital or intracranial extension. The tumor was removed via intranasal endoscopic approach and radiotherapy was administered postoperatively. The patient is currently being followed, and there has been no recurrence in 24 months after surgery. PMID- 12090953 TI - Sublabial approach to sinonasal juvenile ossifying fibroma. AB - Juvenile ossifying fibroma is a rare, unusual fibro-osseous benign tumor of the craniofacial region. It is a progressively growing tumor and has to be removed completely to prevent recurrence. A radical surgery is not advisable in a pediatric patient. Here we describe the sublabial approach for excision of this tumor that allows the complete removal of the tumor without compromising with the aesthetic looks of the patient. PMID- 12090954 TI - Management of sharp esophageal foreign bodies in young children: a cause for worry. AB - The most common foreign bodies in children are coins, particularly in the age group of 2-5 years. Sharp esophageal foreign bodies in children are not commonly encountered. We present a report of two sharp foreign bodies in young children. Both of them required cervical esophagotomy for removal of the foreign body after an unsuccessful endoscopic attempt. The principles of evaluation and management of sharp and penetrating foreign bodies of the esophagus are described. PMID- 12090955 TI - Separation of church and "state". PMID- 12090956 TI - Pregnant adolescents: who agrees to HIV screening, who doesn't. PMID- 12090957 TI - Mexican adolescents and sexually transmitted diseases. a privacy problem. PMID- 12090958 TI - Adolescent girls' perception and experiences with the reality female condom. PMID- 12090959 TI - Internet safety: adolescents' self-report. AB - We examined the association between adolescents' unsafe experience online, types of Internet activity, and safety practices using a questionnaire returned by 213 private school students (seventh through tenth grades) in spring 1999. One-fourth of respondents reported unsafe experiences. Types of unsafe experience varied with gender, Internet activity, and identity sharing. PMID- 12090960 TI - Young adults on the Internet: risk behaviors for sexually transmitted diseases and HIV(1). AB - PURPOSE: To examine the sexual behaviors and related risk factors for sexually transmitted diseases and HIV among young adults who seek sex partners on the Internet. METHODS: Study staff recruited participants in online chat rooms, bulletin boards, and other online venues. A total of 4507 participants responded to a 68-item, self-administered, online survey of Internet sex-seeking practices. The survey solicited information on sexual behavior with partners found on the Internet; in addition, a parallel set of questions addressed sexual behaviors with partners found off the Internet. Of the respondents, 1234 (27.4%) were 18-24 years old. Of the young adults, 61% were male and 75% were white. Responses from young adults were compared to those received from older adults. In addition, responses from young adults who seek sex partners online were compared to responses from young adults who do not seek sex partners online. Analyses, including logistic regression, Chi-square tests, Student's t-tests, and analyses of variance, focused on the difference between young and older adults, as well as the differences in sexual behavior with partners located online and offline. RESULTS: Young adults who seek sex on the Internet report substantially different sexual behavior patterns than young adults who do not seek sex on the Internet. Young adults with online partners reported sexual behaviors similar to older respondents who used the Internet to find sex partners; however, older respondents were more likely than young adults to have been tested for sexually transmitted diseases and HIV. CONCLUSIONS: Young adults who seek sex partners online may be at significantly greater risk for sexually transmitted diseases than their peers who do not seek sex partners online. These data point to an urgent need for online sexual health promotion. PMID- 12090961 TI - Relative power between sexual partners and condom use among adolescents. AB - PURPOSE: To examine how the relative power of adolescent sexual partners in the domain of emotional intimacy is related to condom use. METHODS: Interviewed 228 adolescents who visited an STD clinic in San Francisco. Adolescents were aged 14 19 years, 69% were female, and they were ethnically diverse. We developed a measure of relative power in the domain of emotional intimacy, by adapting five items from existing measures and developing three items ourselves. The partner who had less desire for emotional intimacy was considered to have more power in that domain. We also measured relative decision-making power. Data were analyzed using logistic regression and Student's t-tests. RESULTS: Adolescents who had more power than their partners in the domain of emotional intimacy were more likely to get their way about condom use than adolescents who had less power in this domain. Decision-making power was not related to whether adolescents got their way about condom use. Young men reported greater emotional intimacy power and greater decision-making power than young women. However, gender was not related to getting one's way about condom use. CONCLUSIONS: The results suggest the importance of assessing relative power in the sexual relationships of adolescents when predicting condom use. PMID- 12090962 TI - A randomized comparison of A-CASI and phone interviews to assess STD/HIV-related risk behaviors in teens. AB - PURPOSE: To compare response bias associated with a telephone survey of sexually transmitted disease/human immunodeficiency virus (STD/HIV)-related risk behaviors and an in-home self-administered audio computer assisted self interview (A-CASI). METHODS: We randomly assigned an urban household sample of 223 African-American adolescents to a telephone interview or an A-CASI in their home. The sample was previously recruited by telephone for an earlier study regarding STDs and sexual behavior. We queried participants about their STD/HIV-related risk behaviors. We also assessed their perceived comfort, honesty, and accuracy in answering questions in the different modes through a telephone computer-assisted self interview (T-CASI). RESULTS: There were no significant differences by mode in percentages of participants reporting STD/HIV-related risk behaviors, except more A-CASI participants reported having engaged in sexual intercourse in past 3 months (43.8% vs. 33.3%). There were no differences in perceived comfort, honesty, and accuracy in answering questions in the different modes. These results also did not change after we adjusted for age, household structure, and current school enrollment. CONCLUSIONS: Telephone interviews, a more economical mode, can be employed without much risk of increasing the response bias in the data assessing crude measures of risk. PMID- 12090963 TI - Tapping youth as agents for change: evaluation of a peer leadership HIV/AIDS intervention. AB - PURPOSE: To evaluate the impact of a community-based HIV/AIDS peer leadership prevention program on newly enrolled peer leaders and youth enrolled as peer educators for one or more years (repeat peer leaders). METHODS: Quasi experimental nonrandomized design with two intervention groups (newly enrolled and repeat peer leaders) and one comparison group. The sample consisted of 235 adolescents, 164 peer leaders, and 71 comparison youth, drawn from nine communities in Massachusetts. The intervention consisted of a short course and ongoing group work with an adult advisor to plan and implement HIV/AIDS outreach activities for youth. A confidential questionnaire administered at baseline and postintervention measured change in (a) HIV/AIDS knowledge, (b) planning and presenting skills, (c) self-efficacy, (d) perception of one's self as a change agent in the community, and (e) sexual risk-taking behaviors. Information was collected from both groups of peer leaders on specific activities resulting from the program and perceived benefits. Data were analyzed by both descriptive and multivariate statistics. RESULTS: Over a 9-month period newly enrolled peer leaders had significantly higher mean scores for HIV/AIDS knowledge and perception of one's self as a change agent in the community than comparison youth. On all baseline outcome measures except risk-taking behaviors, repeat peer leaders reported higher scores than newly enrolled peer leaders. Post intervention, HIV/AIDS knowledge continued to increase significantly more among repeat peer leaders compared with those newly enrolled in the program. Repeat peer leaders also reported more benefits from peer leadership training. CONCLUSIONS: A peer education program was found to have benefits to adolescent peer leaders. Benefits gained from the program were sustained and enhanced over time as evidenced by repeat peer leaders included in the study. PMID- 12090964 TI - Mass media as a population-level intervention tool for Chlamydia trachomatis screening: report of a pilot study. AB - PURPOSE: To determine the feasibility and affect of mass media use in a population-level intervention for chlamydia screening promotion. METHODS: A population-level chlamydia intervention protocol was field tested. The intervention, targeting 15-25-year-old individuals, was designed to: (a) increase awareness of personal risk for chlamydial infection; (b) facilitate dissemination of chlamydia knowledge by use of a telephone hot line; and (c) promote care seeking behavior (report for a chlamydia screening program). The intervention activities included: (a) mail outreach, (b) a television and radio campaign, (c) a prerecorded Check-It-Out chlamydia hot line, (d) a staffed chlamydia Options information line, and (e) a free confidential urine ligase chain reaction (LCR) test for chlamydia. Mass mailings were scheduled at intervals, starting two-weeks before the beginning of the television advertisement. The 30-second television advertisement was aired on local television stations 130 times in a 6-week period. The outcome measures were quantity and characteristics of incoming calls to the automated hot line and staffed chlamydia information phone line in response to the chlamydia campaign, and response to the urine screening program. Descriptive and bivariate analyses were used to evaluate the outcomes. RESULTS: The hot line was called 642 times during the monitoring period (November 1, 1999 to March 8, 2000), the majority (92%) during the 6 weeks of television advertisement, with an average of 99 calls per week, compared with an average of 9 calls per week after the commercial ended. Each bulk mailing was accompanied by a boost in the incoming hot line calls. The research staff triaged 133 calls to the "Options" phone line, 81% in the 6 weeks of the TV ad. The mean age of the 133 callers was 23.9 +/- 7.7 years (range 14-49 years). A majority called for screening information; 67% of callers were females and 84% of female callers were under age 26 years. Five percent of callers identified themselves as a parent of a teenager. The majority credited the TV ad as their source of the hot line number. Thirty-one individuals reported for a confidential chlamydia screening, 27 of 31 (87%) during the 6 weeks of TV advertising. No negative responses regarding the chlamydia campaign were encountered. CONCLUSION: This report describes strategies used to implement and measure the effectiveness of a mass media campaign and demonstrates evidence that mass media is effective in delivering STD intervention messages to young people. PMID- 12090965 TI - Sexual behaviors of physically disabled adolescents in the United States. AB - PURPOSE: To examine differences in some aspects of sexual behavior between physically disabled and nondisabled adolescents in the United States. METHODS: The 1994-1995 Wave I data from the National Longitudinal Study of Adolescent Health (Add Health), a stratified, multistage cluster, school-based sample of 7th to 12th graders, were used. Physical disabilities were measured in terms of functional limitation and activity restriction. A total of 1153 adolescents were identified as physically disabled, mainly with limb difficulties. We studied their exposure on school sex education, contraceptive knowledge, sex attitudes, and popularity. Their experiences in romantic attraction and sex were analyzed by multinomial logistic regression methods, controlling for age, pubertal development, and other sociodemographic variables. RESULTS: Although disabled adolescents are slower in pubertal development and more socially isolated, they are as sexually experienced as their nondisabled counterparts. Their exposure to school sex education is largely similar to that of the nondisabled, yet disabled boys are somewhat less knowledgeable about birth control. Severely disabled boys who are sexually active are popular among their same-sex peers, much like nondisabled boys. Although disabled adolescents are as sexually experienced, severely disabled boys and girls have less opposite-sex romantic attraction and are less sure about their sexual preference. Adolescents with mild disabilities have a higher tendency for same-sex attraction than do the nondisabled. Adolescent girls with physical disabilities consistently have higher odds of experiencing forced sex. CONCLUSIONS: Physically disabled adolescents' sexual development and their specific needs for sex education and guidance should not be overlooked. More attention should be paid to protect physically disabled girls from sexual abuses and exploitations. PMID- 12090966 TI - A longitudinal study of maternal messages about dating and sexuality and their influence on Latino adolescents. AB - PURPOSE: To learn more about the nature of mother-adolescent discussions about sexuality and their influence on adolescents' behaviors and attitudes in Latino families. METHODS: Fifty-five Latino mothers and their midadolescent children (35 girls, 20 boys, mean age 13 years) were observed talking about dating and sexuality. The videotaped conversations were coded for the prevalence of five maternal message types: beliefs and values, advice, cautionary messages, comments about the adolescents' everyday experiences, and maternal self-disclosure about her own dating and sexuality experiences. A structural equation model was used to examined how time spent on the various message types was related to three adolescent-related outcomes collected about 1 year later: sexual behavior, attitudes toward premarital sex, and reports of openness in the mother-child relationship. RESULTS: Latino mothers focused the conversation on their personal beliefs and values about dating and sexuality. Discussions about beliefs and values were accompanied by more parental advice and cautionary messages. More maternal self-disclosure about personal dating and sexuality experiences was related to the adolescents' reporting a better functioning relationship with their mothers and having more conservative attitudes toward premarital sex. Sexuality communication was both positively and negatively related to adolescent sexual behavior, but the messages were not the same. CONCLUSIONS: Maternal communication in Latino families influences adolescents' behaviors and attitudes toward sex and adolescents' perceptions of openness in the mother-child relationship. Observational methods are useful for studying the complexity of the communication process and for studying participants from ethnic minority populations who may be unfamiliar with or uncomfortable filling out questionnaires. PMID- 12090967 TI - Keeping middle school students abstinent: outcomes of a primary prevention intervention. AB - PURPOSE: (a) To test the longer-term (6-12 month) effect of a school-based intervention designed to delay the onset of sexual intercourse on continuation of abstinence, (b) to compare the effect of the intervention when delivered by different providers, and (c) to describe the factors that influence students' transition from abstinence to sexual activity. METHODS: This study was a nonrandomized control trial with one control and three intervention groups. The setting was health education classes in urban, predominantly ethnic minority schools. The participants were middle school students (N = 1352; mean age, 13.1 years) in five schools. Of participants, 50% were African American, 20% white, 16% Hispanic, and 14% other. Youth were assigned to one of four groups. The control group consisted of the regular school health curriculum and teacher. All three intervention groups received the Rochester AIDS Prevention Project curriculum, but implemented by different types of instructors, including ethnically diverse male-female pairs of adult professional educators; male-female pairs of extensively trained high school peer educators; and school district health teachers. A confidential questionnaire administered preintervention and at long-term follow-up (mean, 44 weeks) measured demographics, risk behaviors, and sexual intercourse history. RESULTS: At preintervention, 27% of girls and 62% of boys reported sexual intercourse experience. At follow-up, 19% and 32%, respectively, of the previously abstinent girls and boys had "transitioned" to sexual activity. Increasing age (p <.01, females; p <.001, males), lower socioeconomic status (p <.0001), and higher general risk behaviors (p <.0001) best predicted the transition. Logistic regression indicated that the intervention was effective for peer-taught males (p =.02) and regular teacher taught males (p =.001) and females (p =.05). CONCLUSIONS: Successful abstinence maintenance was only possible among those subjects who were not already sexually experienced at study enrollment. Baseline scores regarding intercourse and general life risks already evident by seventh grade suggest that urban, school based primary prevention interventions must occur before adolescence. Early adolescence interventions need to include both abstinence and safer sex messages. PMID- 12090968 TI - Coital status and perceptions about sexual abstinence refusal skills. AB - PURPOSE: To compare perceptions and use of sexual refusal skills. METHODS: A voluntary, anonymous sample of ninth and tenth grade students (N = 2256) reported their perceptions about and use of sexual refusal skills that had been taught in school. Forty-four percent of the students classified themselves as virgins, 18% reported one sexual partner, and 38% reported multiple sex partners. Coital status groups were compared on measures addressing refusal skills using Chi square tests. RESULTS: Approximately one-third of the students reported using refusal skills either a lot or a little, with virgins more likely to indicate that they used refusal skills a lot. Virgins also were least likely to say that they should have used the refusal skills or that they didn't feel like using refusal skills. Furthermore, virgins were most likely to state that they didn't need to use refusal skills. One partner and multiple partner groups were more likely to indicate that they had not been taught refusal skills. CONCLUSIONS: The majority of students did not use the refusal skills taught to them in school; however, all coital status groups used the refusal skills in some capacity. Differences between the virgin and sexually experienced groups present challenges for instructors and curriculum developers. Some of the commonly held assumptions about refusal skill curricula may not be true. PMID- 12090969 TI - Risk assessment of adolescents with same-sex relationships. AB - PURPOSE: To compare the risk status on health and behavior for those with same sex partners and those without. METHODS: Add Health data provide a sample of 20,745 adolescents in grades 7 through 12 interviewed at home. The risk statuses of respondents with no partners, same-sex-only partners, and partners of both sexes were compared to respondents with opposite-sex partners only. Respondents were evaluated on selected personal and social attributes (verbal IQ, family structure, masculinity, popularity), and risk status (substance use, depression, suicidal thoughts, anal sex, general delinquency, being physically attacked, perceived risk of being killed or getting AIDS). Data were analyzed by logistic and linear regression using STATA to adjust for clustering and sampling weights. RESULTS: Compared to boys with opposite-sex-only partners, boys with same-sex only partners were at high risk for emotional problems, but not delinquency or substance use. Boys with partners of both sexes were at high risk for delinquency and substance use, but not for emotional problems. Neither group of boys with same-sex partners is at high risk of being attacked compared to those with opposite-sex partners only. Girls with only same-sex partners are never a high risk group, while girls with partners of both sexes are the high-risk category in every case. CONCLUSIONS: Adolescents with same-sex-only partners do not resemble those with partners of both sexes in risk status. Combining the two categories obscures the unique risk profile of those with both-sex partners, and obscures the low risk on most variables but the high emotional risk of boys with only same sex partners. PMID- 12090970 TI - Long-term reductions in sexual initiation and sexual activity among urban middle schoolers in the reach for health service learning program. AB - PURPOSE: To evaluate the sustained effectiveness of a middle school service learning intervention on reducing sexual initiation and recent sex among urban African-American and Latino adolescents from 7th grade through the 10th grade. METHODS: During the fall of seventh grade and again in eighth grade, students were randomly assigned by classroom to participate either in community youth service (CYS) or not (controls). Service learning is an educational strategy that couples meaningful service in the community with classroom instruction. Students in both intervention and control conditions received classroom health lessons. Surveys were conducted at seventh grade baseline and at the end of 10th grade, approximately 2 years after intervention. Self-reported sexual behaviors of youths who had participated in CYS were compared with those of controls receiving classroom curriculum alone (n = 195). RESULTS: CYS participants were significantly less likely than controls to report sexual initiation (2 years CYS, odds ratio [OR] = 0.32; 1 year, OR = 0.49) as well as recent sex (2 years CYS, OR = 0.39; 1 year CYS, OR = 0.48). Among those who were virgins at seventh grade, 80% of males in the curriculum-only condition had initiated sex, compared with 61.5% who received 1 year of CYS, and 50% who received 2 years. Among females, the figures were 65.2%, 48.3%, and 39.6%, respectively. CONCLUSION: A service learning intervention that combines community involvement with health instruction can have a long-term benefit by reducing sexual risk taking among urban adolescents. PMID- 12090971 TI - Emergency contraception among teenagers in Switzerland: a cross-sectional survey on the sexuality of 16- to 20-year-olds. AB - PURPOSE: To describe and analyze emergency contraception (EC) awareness and use among sexually active Swiss teenagers. METHODS: Anonymous computerized questionnaires were distributed to a national representative sample of 4283 in school adolescents (aged 16 to 20 years) in high schools and professional centers. Young people who were sexually active (51.5% of the sample: 1058 girls and 1073 boys) responded to questions on EC awareness and use and on sexual perception, attitude, and behaviors. Univariate analyses and multiple regression analyses were used to describe EC awareness and use and their correlates. RESULTS: Most of the sexually active girls (89.3%) and boys (75.2%) knew of the existence of EC. Of girls, 20% reported having used EC, and the majority of them used it only once (64.1%) or twice (18.5%). EC awareness was positively associated with the father's level of education (girls: odd ratio 5.18) and the scholastic curriculum of the respondent. Gender differences in the correlates of EC awareness demonstrate that girls who had a confidant or a group of friends or boys of Swiss nationality and those who have had the opportunity to discuss the issue of contraception declare greater awareness of EC. EC use was higher among girls who lived in urban areas (odds ratio 1.91) and occasionally had unprotected intercourse. We did not find any significant difference in the profile of multiple vs. one-time users. CONCLUSION: EC awareness and use should be improved through better information and accessibility, especially among teenagers who place themselves in at-risk situations. PMID- 12090972 TI - Beyond trading insults in international humanitarian aid. PMID- 12090973 TI - Aromatase inhibitors for the endocrine adjuvant treatment of breast cancer. PMID- 12090974 TI - Glucose-tolerance testing in acute myocardial infarction. PMID- 12090975 TI - Responding to fear of childbirth. PMID- 12090976 TI - Economic implications of non-compliance in health care. PMID- 12090977 TI - Anastrozole alone or in combination with tamoxifen versus tamoxifen alone for adjuvant treatment of postmenopausal women with early breast cancer: first results of the ATAC randomised trial. AB - BACKGROUND: In the adjuvant setting, tamoxifen is the established treatment for postmenopausal women with hormone-sensitive breast cancer. However, it is associated with several side-effects including endometrial cancer and thromboembolic disorders. We aimed to compare the safety and efficacy outcomes of tamoxifen with those of anastrozole alone and the combination of anastrozole plus tamoxifen for 5 years. METHODS: Participants were postmenopausal patients with invasive operable breast cancer who had completed primary therapy and were eligible to receive adjuvant hormonal therapy. The primary endpoints were disease free survival and occurrence of adverse events. Analysis for efficacy was by intention to treat. FINDINGS: 9366 patients were recruited, of whom 3125 were randomly assigned anastrozole, 3116 tamoxifen, and 3125 combination. Median follow-up was 33.3 months. 7839 (84%) patients were known to be hormone-receptor positive. Disease-free survival at 3 years was 89.4% on anastrozole and 87.4% on tamoxifen (hazard ratio 0.83 [95% CI 0.71-0.96], p=0.013). Results with the combination were not significantly different from those with tamoxifen alone (87.2%, 1.02 [0.89-1.18], p=0.8). The improvement in disease-free survival with anastrozole was seen in the subgroup of hormone-receptor-positive patients, but not the receptor-negative patients. Incidence of contralateral breast cancer was significantly lower with anastrozole than with tamoxifen (odds ratio 0.42 [0.22 0.79], p=0.007). Anastrozole was significantly better tolerated than tamoxifen with respect to endometrial cancer (p=0.02), vaginal bleeding and discharge (p<0.0001 for both), cerebrovascular events (p=0.0006), venous thromboembolic events (p=0.0006), and hot flushes (p<0.0001). Tamoxifen was significantly better tolerated than anastrozole with respect to musculoskeletal disorders and fractures (p<0.0001 for both). INTERPRETATION: Anastrozole is an effective and well tolerated endocrine option for the treatment of postmenopausal patients with hormone-sensitive early breast cancer. Longer follow-up is required before a final benefit:risk assessment can be made. PMID- 12090978 TI - Glucose metabolism in patients with acute myocardial infarction and no previous diagnosis of diabetes mellitus: a prospective study. AB - BACKGROUND: Glycometabolic state at hospital admission is an important risk marker for long-term mortality in patients with acute myocardial infarction, whether or not they have known diabetes mellitus. Our aim was to ascertain the prevalence of impaired glucose metabolism in patients without diagnosed diabetes but with myocardial infarction, and to assess whether such abnormalities can be identified in the early course of a myocardial infarction. METHODS: We did a prospective study, in which we enrolled 181 consecutive patients admitted to the coronary care units of two hospitals in Sweden with acute myocardial infarction, no diagnosis of diabetes, and a blood glucose concentration of less than 11.1 mmol/L. We recorded glucose concentrations during the hospital stay, and did standardised oral glucose tolerance tests with 75 g of glucose at discharge and again 3 months later. FINDINGS: The mean age of our cohort was 63.5 years (SD 9) and the mean blood glucose concentration at admission was 6.5 mmol/L (1.4). The mean 2-h postload blood glucose concentration was 9.2 mmol/L (2.9) at hospital discharge, and 9.0 mmol/L (3.0) 3 months later. 58 of 164 (35%, 95% CI 28-43) and 58 of 144 (40%, 32-48) individuals had impaired glucose tolerance at discharge and after 3 months, respectively, and 51 of 164 (31%, 24-38) and 36 of 144 (25%, 18-32) had previously undiagnosed diabetes mellitus. Independent predictors of abnormal glucose tolerance at 3 months were concentrations of HbA(1c) at admission (p=0.024) and fasting blood glucose concentrations on day 4 (p=0.044). INTERPRETATION: Previously undiagnosed diabetes and impaired glucose tolerance are common in patients with an acute myocardial infarction. These abnormalities can be detected early in the postinfarction period. Our results suggest that fasting and postchallenge hyperglycaemia in the early phase of an acute myocardial infarction could be used as early markers of high-risk individuals. PMID- 12090979 TI - Safety and efficacy of transfusions of autologous cryopreserved platelets derived from recombinant human thrombopoietin to support chemotherapy-associated severe thrombocytopenia: a randomised cross-over study. AB - BACKGROUND: The increasing demand for platelet products, and concern over the transfusion-associated risks of alloimmunisation and infections, have motivated a search for improved methods aimed at keeping exposure to donor antigens to a minimum. Transfusion of thrombopoietin-derived autologous platelets might provide an alternative strategy. We aimed to compare the safety and efficacy of this strategy with that of transfusion with fresh allogeneic platelets in patients with severe chemotherapy-induced thrombocytopenia. METHODS: 20 patients with gynaecological malignancies were treated with two doses of 1.2 microg/kg recombinant human thrombopoietin. From day 12, we aimed to collect 50 units of platelets from these patients by plateletpheresis. Harvested platelets were cryopreserved in ThromboSol and 2% dimethyl sulfoxide (DMSO) for use in subsequent autologous transfusions. Patients then received carboplatin for up to six cycles. Patients were randomly assigned to group A (n=10), which received allogeneic fresh platelets at the first instance of severe thrombocytopenia (platelet count <15,000/microL) and then autologous cryopreserved platelets at the next, or to group B (n=10), which received first autologous and then allogeneic platelets. In subsequent cycles, all patients received autologous platelets while available. The primary endpoint was platelet count increment corrected for the number of platelets transfused and the patients' body-surface area. Analysis was by intention to treat. FINDINGS: Treatment with recombinant human thrombopoietin significantly increased platelet count (median 2.3-fold [range 1.5-3.3], p<0.0001) in all but one patient in group A. The median number of platelets collected per patient was 53 units (14-66) in two collections (one to three). There was no significant difference in the corrected platelet count increments (CCIs) between the 19 paired transfusions of cryopreserved autologous platelets and fresh allogeneic platelets (median 1-h CCI 15.7 vs 19.8, p=0.398; median 24-h CCI 13.0 vs 18.1, p=0.398). 14 of the 19 patients had a good response (1-h CCI >7.5) to their first transfusion of allogeneic platelets. By contrast, all patients had a good response to their first transfusion of autologous platelets (p=0.063). Moreover, no significant decrease in the CCIs (p=0.405) was seen over six cycles after autologous platelet transfusions (n=63). No transfusion reactions or any serious adverse event was recorded during autologous platelet transfusions. INTERPRETATION: Recombinant human thrombopoietin facilitated collection of multiple units of platelets, which could be cryopreserved and reinfused to counteract severe thrombocytopenia during multicycle chemotherapy. Transfusion of autologous cryopreserved platelets derived from recombinant human thrombopoietin can provide a viable strategy to minimise the risks of allogeneic platelet transfusions and provide a long-lasting supply of platelet support. PMID- 12090980 TI - Management of suspected acute viral upper respiratory tract infection in children with intranasal sodium cromoglicate: a randomised controlled trial. AB - BACKGROUND: Upper respiratory tract infection in children is one of the most frequent reasons for visiting a family doctor, and antibiotics are often prescribed inappropriately. Sodium cromoglicate inhibits the ICAM-1 molecule, which is the receptor for human rhinoviruses. We aimed to investigate whether intranasal cromoglicate shortens duration of infection of the upper respiratory tract. METHODS: We randomly assigned 290 children diagnosed with suspected acute viral upper respiratory tract infection by their family doctor (137 boys, 153 girls; mean age 5.2 years [SD 3.39]) either intranasal 4% sodium cromoglicate spray or intranasal normal saline spray. Follow-up was by daily symptom diary for 2 weeks and by telephone. Canadian Acute Respiratory Illness and Flu Scale (CARIFS) score was the primary outcome measure. FINDINGS: 195 patients returned symptom diaries, and 20 of these could not be included in the main analysis. 246 patients completed the telephone interview at week 1. There was no difference in recovery rate over the first week between the two groups, with the estimated difference in slope of log (CARIFS) being -0.01 (95% CI -0.05 to 0.03). There were no differences between the two groups in side-effects or re-consultation rates. 43 (17%) of 246 children with suspected acute viral upper respiratory tract infection went back to see their family doctor, and 220 (89%) of 246 were managed without prescription of antibiotics. INTERPRETATION: Intranasal sodium cromoglicate is not a useful additional treatment for this infection. Our results further clarify the role of prescribed drugs for children with these frequent illnesses. PMID- 12090981 TI - Pulmonary tumour embolism. PMID- 12090982 TI - Concentration in plasma of macrophage inhibitory cytokine-1 and risk of cardiovascular events in women: a nested case-control study. AB - BACKGROUND: Macrophage inhibitory cytokine-1 (MIC-1) is part of the TGF-beta superfamily. Raised concentrations of MIC-1 in serum arise in several disease states, can be detected in normal individuals, and can partly be genetically determined. We aimed to investigate whether MIC-1 has a role in atherothrombosis. METHODS: We did a prospective, nested, case-control study in 27628 initially healthy women. Of these women, we established baseline concentrations of MIC-1 in 257 who subsequently had myocardial infarction, stroke, or died from a cardiovascular event (cases) and in 257 matched for age and smoking status, who did not report cardiovascular disease during 4-year follow-up (controls). We also assessed polymorphisms in the MIC-1 gene (MIC-1 H and MIC-1 D) in all 514 women. FINDINGS: MIC-1 concentrations were higher at baseline in women who subsequently had cardiovascular events than in those who did not (618 vs 538 pg/mL, p=0.0002). Concentrations above the 90th percentile (>856 pg/mL) were associated with a 2.7 fold increase in risk (95% CI 1.6-4.9, p=0.001). This effect was independent of traditional cardiovascular risk factors and at least additive to that of C reactive protein. There was no significant association between MIC-1 polymorphism and vascular events. INTERPRETATION: MIC-1 could be a novel target for cardiovascular disease prevention. PMID- 12090983 TI - Aphasia in a farmer following viper bite. PMID- 12090984 TI - Outcome of twin pregnancies with complete hydatidiform mole and healthy co-twin. AB - We assessed 77 twin pregnancies, comprising complete hydatidiform mole (CHM) and healthy co-twin, to ascertain the risks to the mother and baby of continuing the pregnancy, versus termination. 24 women with histologically confirmed CHM and healthy co-twin pregnancies decided to have a termination. 53 women continued with their pregnancies, though two had to have terminations because of severe pre eclampsia, and 23 spontaneously aborted (<24 weeks' gestation). 28 pregnancies lasted 24 weeks or more, resulting in 20 livebirths. Chemotherapy to eliminate persistent gestational trophoblastic disease (pGTD) was required in three of 19 women (16%; 95% CI 3-39) who terminated their pregnancies in the first trimester, and in 12 of 58 (21%; 95% CI 11-33%) who continued their pregnancies. CHM and healthy co-twin pregnancies have a high risk of spontaneous abortion, but about 40% result in livebirths, without significantly increasing the risk of pGTD. PMID- 12090985 TI - Diagnosis of vascular graft infections with antibodies against staphylococcal slime antigens. AB - Late-onset infections of synthetic vascular grafts (LO-SVGIs) are generally caused by staphylococci that produce a slime polysaccharide and grow as a biofilm on the graft surface. We developed an ELISA to detect serum antibodies against staphylococcal slime polysaccharide antigens (SSPA). Patients with an ongoing staphylococcal LO-SVGI had greater titres of IgM antibodies against SSPA than did patients in other groups. Antibody titres of 0.40 ELISA units (EU) or more, or 0.35 EU or more detected 97% and 100% of staphylococcal LO-SVGIs, respectively, 0% and 2% titre/unit false-positive results. Our findings suggest that such an ELISA represents a sensitive, specific, and non-invasive diagnostic test for staphylococcal LO-SVGIs. PMID- 12090987 TI - Two new studies likely to add fire to stem-cell debate. PMID- 12090988 TI - Update on islet-cell transplantation for type 1 diabetes. PMID- 12090986 TI - Association between aplastic anaemia and mutations in telomerase RNA. AB - The main cause of aplastic anaemia remains elusive. Germline mutations in the gene encoding the RNA component of telomerase (hTR) have been seen in the autosomal dominant form of dyskeratosis congenita--an inherited syndrome characterised by aplastic anaemia. By screening the hTR gene, we identified mutations in two of 17 patients with idiopathic aplastic anaemia, three of 27 patients with constitutional aplastic anaemia, but in none of 214 normal controls (p<0.0001). Furthermore, patients with hTR mutations had significantly shorter telomeres than age-matched controls (p=0.027). These data indicate that, in a subset of patients with aplastic anaemia, the disorder might be associated with a genetic lesion in the telomere maintenance pathway. PMID- 12090993 TI - Children targeted in research-awareness campaign. PMID- 12090994 TI - Largest ever measles vaccination programme launched in Africa. PMID- 12090995 TI - Where have Canada's general practitioners gone? PMID- 12090996 TI - UK's new consultant contract finalised. PMID- 12090997 TI - HIV-1/AIDS and the control of other infectious diseases in Africa. AB - The effect of HIV-1 on other infectious diseases in Africa is an increasing public health concern. In this review, we describe the role that three major infectious diseases--malaria, sexually transmitted diseases (STDs), and tuberculosis--have had in the HIV-1 epidemic. The high prevalence of untreated STD infections has been a major factor facilitating the spread of HIV-1 in Africa; with the synergistic interaction between HIV-1 transmission and genital herpes being of special concern for control of both diseases. Increased susceptibility to tuberculosis after infection with HIV-1 has led to a rising incidence and threat of increased transmission of tuberculosis. Clinical malaria occurs with an increased frequency and severity in HIV-1-infected individuals, especially during pregnancy. As with tuberculosis, STDs, and other communicable HIV-1-associated diseases, the net effect of HIV-1 might include increased rates of malaria transmission across communities. In addition to enhancing access to HIV-1 prevention and care, public health surveillance and control programmes should be greatly intensified to cope with the new realities of infectious disease control in Africa. PMID- 12090998 TI - Drug development for neglected diseases: a deficient market and a public-health policy failure. AB - There is a lack of effective, safe, and affordable pharmaceuticals to control infectious diseases that cause high mortality and morbidity among poor people in the developing world. We analysed outcomes of pharmaceutical research and development over the past 25 years, and reviewed current public and private initiatives aimed at correcting the imbalance in research and development that leaves diseases that occur predominantly in the developing world largely unaddressed. We compiled data by searches of Medline and databases of the US Food and Drug Administration and the European Agency for the Evaluation of Medicinal Products, and reviewed current public and private initiatives through an analysis of recently published studies. We found that, of 1393 new chemical entities marketed between 1975 and 1999, only 16 were for tropical diseases and tuberculosis. There is a 13-fold greater chance of a drug being brought to market for central-nervous-system disorders or cancer than for a neglected disease. The pharmaceutical industry argues that research and development is too costly and risky to invest in low-return neglected diseases, and public and private initiatives have tried to overcome this market limitation through incentive packages and public-private partnerships. The lack of drug research and development for "non-profitable" infectious diseases will require new strategies. No sustainable solution will result for diseases that predominantly affect poor people in the South without the establishment of an international pharmaceutical policy for all neglected diseases. Private-sector research obligations should be explored, and a public-sector not-for-profit research and development capacity promoted. PMID- 12090999 TI - The tip of the iceberg? PMID- 12091000 TI - Disparity between angiographic regression and clinical event rates with hydrophobic statins. PMID- 12091001 TI - Losartan for cardiovascular disease in patients with and without diabetes in the LIFE study. PMID- 12091002 TI - Losartan for cardiovascular disease in patient's with and without diabetes in the LIFE study. PMID- 12091003 TI - Losartan for cardiovascular disease in patient's with and without diabetes in the LIFE study. PMID- 12091004 TI - Losartan for cardiovascular disease in patient's with and without diabetes in the LIFE study. PMID- 12091005 TI - Losartan for cardiovascular disease in patient's with and without diabetes in the LIFE study. PMID- 12091006 TI - Losartan for cardiovascular disease in patient's with and without diabetes in the LIFE study. PMID- 12091007 TI - Losartan for cardiovascular disease in patient's with and without diabetes in the LIFE study. PMID- 12091008 TI - Losartan for cardiovascular disease in patient's with and without diabetes in the LIFE study. PMID- 12091009 TI - Losartan for cardiovascular disease in patient's with and without diabetes in the LIFE study. PMID- 12091010 TI - Losartan for cardiovascular disease in patient's with and without diabetes in the LIFE study. PMID- 12091012 TI - Outcomes in pregnancy. PMID- 12091013 TI - Outcomes in pregnancy. PMID- 12091014 TI - Shortening inpatient stay for stroke. PMID- 12091015 TI - Twins and fetal origins hypothesis: within-pair analyses. PMID- 12091016 TI - Neurosurgery 7000 years ago in central Italy. PMID- 12091024 TI - The pathophysiology and management of bowel and mesenteric injuries due to blunt trauma. PMID- 12091025 TI - The hand-held FAST: experience with hand-held trauma sonography in a level-I urban trauma center. AB - AIMS: To describe the effectiveness of a portable hand-held ultrasound machine when used by clinicians in the early evaluation and resuscitation of trauma victims. METHODS: The study was a prospective evaluation in a level-I urban trauma center. The focussed assessment with sonography for trauma is a specifically defined examination for free fluid known as the focused assessment with sonography for trauma (FAST) exam. Seventy-one patients had a hand-held FAST (HHFAST) examination performed with a Sonosite 180, 2.4 kg ultrasound machine. Sixty-seven examinations were immediately repeated with a Toshiba SSH 140A portable floor-based machine. This repeat scan (formal FAST or FFAST) was used as a comparison standard between the devices for study purposes. Four patients had a HHFAST only, all with positive result, two being taken for immediate laparotomy, and two having a follow-up computed tomographic (CT) scan. Patient follow-up from other imaging studies, operative intervention, and clinical outcomes were also compared to the performance of each device. RESULTS: There were 58 victims of blunt, and 13 of penetrating abdominal trauma. One examination was indeterminate using both machines. The apparent HHFAST performance yielded; sensitivity, specificity, positive predictive value, negative predictable value, and accuracy (S, S, PPV, NPV, A) of 83, 100, 100, 98, 98%. Upon review, a CT scan finding and benign clinical course found the HHFAST diagnosis to be correct rather than the FFAST in one case. Considering the ultimate clinical course of the patients, yielded a (S, S, PPV, NPV, A) of 78, 100, 100, 97, and 97% for the HHFAST and 75, 98, 86, 97, and 96% for the FFAST. Statistically, there was no significant difference in the actual performance of the HHFAST compared to the FFAST in this clinical setting. DISCUSSION: Hand-held portable sonography can simplify early and accurate performance of FAST exams in victims of abdominal trauma. PMID- 12091026 TI - A multidisciplinary approach in the management of hepatic injuries. AB - We reviewed 87 patients with hepatic injuries who were admitted to King Chulalongkorn Memorial Hospital, Bangkok, Thailand, from January 1995 to December 1999; 76% of them had sustained blunt trauma and 24% penetrating trauma. Their injury severity scores (ISS) ranged from 4 to 57 (mean 20.94+/-12.8); 50% of them were in shock on arrival; 8.1, 28.7, 25.3, 19.5, and 18.4% suffered from hepatic injuries graded I, II, III, IV, and V, respectively. Seventeen patients (19.5%) were successfully managed non-operatively; three of them underwent hepatic angiography, which in two revealed leakage of contrast medium from the right hepatic artery; both were successfully treated by embolization. One patient had bile leakage and collection, which was successfully treated by ultrasound-guided percutaneous drainage. Seventy patients (80.5%) underwent exploratory laparotomy; nine of them died in the operating room. Of the remaining 61 who left the operating room alive, 21 had perihepatic packing, which was frequently used in those with injuries to segments V, VI, VII, and VIII (Couinaud's nomenclature). Eight patients who had packing and one who had not died in the postoperative period. Two patients who had packing underwent subsequent hepatic angiography with embolization before successful pack removal. The overall mortality was 20.7%. The mortality in complex hepatic injuries (grades IV and V) was 13 out of 33 (39.4%). We believe that non-operative management should be considered in haemodynamically stable patients. Angiography with embolization is invaluable in improving outcome in both non-operative and operative patients. Perihepatic packing is life-saving in complex hepatic injuries that cannot be effectively treated by simple surgical procedures. Finally, ultrasound- or CT-guided percutaneous drainage of bile leakage or collections spared a number of patients from open and complicated surgery. PMID- 12091027 TI - Critical decisions in trauma of the thoracic aorta. AB - Trauma to the thoracic aorta is a significant injury with a high mortality. This mortality is partly due to the delayed recognition of the condition. The increase of motor vehicle accidents and the use of firearms worldwide has resulted in an increase of the incidence of trauma to the thoracic aorta. Certain new developments in diagnosis and management have resulted in traditional methods being questioned. This review analyses current literature and data which we attempt to evaluate. PMID- 12091029 TI - Efficacy of pre-hospital interventions for the management of severe blunt head injury. PMID- 12091028 TI - Ketamine in war/tropical surgery (a final tribute to the racemic mixture). AB - A technique of continuous intravenous anaesthesia with ketamine was used successfully during the Somalia civil war in 1994 and in north Uganda in 1999 for 64 operations in 62 patients, aged from 6 weeks to 70 years, undergoing limb and abdominal surgery including caesarian sections and interventions in neonates. Operations lasting up to 2h could be performed in the absence of sophisticated equipment such as pulse oximeters or ventilators in patients on spontaneous ventilation breathing air/oxygen only. After premedication with diazepam, glycopyrrolate and local anaesthesia, and induction with standard doses of ketamine, a maintenance dose of 10-20 microg/kg/min of ketamine proved safe and effective. Emphasis was placed on bedside clinical monitoring, relying heavily on the heart rate. Diazepam, unless contraindicated or risky, remains the only necessary complementary drug to ketamine as it buffers its cardiovascular response and decreases the duration and intensity of operative and postoperative hallucinations. Local anaesthetic blocks were useful in decreasing the requirement for postoperative analgesia. An antisialogue was usually unnecessary in operations lasting up to 2 h, glycopyrrolate being the best choice for its lowest psychotropic and chronotropic effects, especially in a hot climate. Experience in war/tropical settings suggests this technique could be useful in civilian contexts such as outdoor life-saving emergency surgery or in mass casualties where, e.g. amputation and rapid extrication were required. PMID- 12091030 TI - The relationship between blood glucose, mean arterial pressure and outcome after severe head injury: an observational study. AB - BACKGROUND AND METHOD: Hypotension and hyperglycaemia occurring in the first 24h after severe head injury are individually associated with poor outcome but a causal effect has not been proven for either. Their combined effect is unknown and is investigated in this observational study of 338 patients with head injury, a Glasgow coma score (GCS) of 8 or less and requiring mechanical ventilation. RESULTS: Mean arterial pressure (MAP) and blood glucose are linearly related to mortality (P<0.0001). Regression analysis shows that each has an independent effect. Moreover, the relationship between blood glucose and mortality is stronger than the relationship between MAP and mortality. When patients are grouped according to lowest MAP, hyperglycaemia is associated with increasing mortality within each group (P<0.0001). CONCLUSION: Further studies on the combined effect of hyperglycaemia and hypotension on mortality after head injury are needed because this study suggests, but does not prove, an additive, causal association. PMID- 12091031 TI - Chronic subdural haematoma in young and extremely aged adults: a comparative study of two age groups. AB - To compare the clinical presentation, etiological factors and the outcome of chronic subdural haematoma (CSDH) in young and old adults, a retrospective analysis was performed by differentiating young adults, age<40 years (n=24) versus extremely aged adults, age>75 years (n=51). The clinical data, computed tomography (CT) findings and surgical outcome were recorded for analysis. We observed certain characteristics particular to young CSDH patients, higher incidence of headache (P<0.001) and vomiting (P=0.009), the shorter duration from trauma to operation (P=0.018) and more severe traumatic episodes (P=0.001). The older patients had a higher frequency of mental change (P=0.006), motor deficits (P=0.014) and larger haematomas (P<0.001). The surgical complication rate was not statistically different between the two age groups. An understanding of the varied clinical presentation of CSDH is essential to stimulate clinical suspicion and prompt evaluation, particularly for the differences between young and extremely aged patients. PMID- 12091032 TI - A retrospective review of emergency admission for head injury in the over 75s. AB - This study retrospectively reviewed all patients aged 75 years and over who were admitted to hospital following an acute head injury. All patients were cared for by general surgeons. The study period lasted for 2 years. Fifty five patients with a mean age of 83.8 years (range 75-97) were admitted. Four patients had a Glasgow Coma Score (GCS) of <13, all had intracranial injuries, only one being fit for neurosurgical intervention. Fifty one patients had a GCS greater than or equal to 13, 25 (49%) were admitted for social reasons, 10 (19.6%) for observation, 9 (17.6%) due to confusion and 7 (13.7%) due to other injuries. Patients spent a total of 366 days in surgical beds. Twenty five patients were discharged or transferred within 24h of admission and 30 patients remained in hospital for a total of 343 days, therefore, 54% of patients occupy 94% of the time spent in hospital. Of these 160 days were spent awaiting transfer to another hospital, 147 days waiting for social services, 24 days waiting for a terminal outcome and 12 days for unspecified reasons. Only 11 (38%) patients who suffered a fall of unknown cause underwent any follow-up investigations. Elderly patients with head injuries rarely require neurosurgical intervention. They often have co existing medical conditions and complex social needs leading to prolonged hospital admission. The management of these patients might be more appropriately undertaken by physicians specializing in elderly care medicine. PMID- 12091033 TI - Prophylaxis of thromboembolism in spinal injuries--survey of practice in spinal units in the British Isles. AB - AIM: Survey and discussion of the current thromboembolic prophylaxis practice in spinal injury units within the British Isles. BACKGROUND: Deep vein thrombosis and pulmonary embolism are major causes of morbidity and mortality in patients with spinal injuries. A wide range of thromboprophylactic measures have been proposed. The optimum treatment and duration for thromboprophylaxis in spinal injuries is unknown. Different spinal units within the British Isles use different measures despite similar training backgrounds. The present study was performed to review practice and make suggestions on best practice using literature review. METHODS: All the 13 regional and national spinal injury referral centres within the British Isles were contacted to find out their protocols for thromboembolic prophylaxis in patients with acute spinal injuries. RESULTS: All units replied. A wide variation in methods used was found in different spinal units ranging from no chemical prophylaxis to oral anticoagulation with warfarin and contrasting views on the use of antithromboembolic stockings. CONCLUSION: All units recognised the risk of thromboembolism after spinal injuries. A wide variety of thromboprophylactic measures are used. PMID- 12091034 TI - Long-term results after posterior fixation of thoraco-lumbar burst fractures. AB - Current concepts of treating thoraco-lumbar burst-compression injuries are based on posterior transpedicular fixation techniques which are angular stable. However, the long-term results of this approach are controversial due to inconsistent reports and due to a paucity of data on late outcome. In the present study we analyzed 50 patients retrospectively who had an unstable burst compression injury at T 11-L 2 (type A 3 according to Magerl) without a neurological deficit. All fractures were stabilized by an internal fixator either with or without transpedicular spongiosa grafting. Patients were treated between 1991 and 1997. Follow-up times ranged from 36 to 103 months. Follow-up examinations collected occupational, subjective and clinical data (activity score, Hannover spine score) and included radiographic measurements. The latter were used to calculate the sagittal index (SI) which measures deformities of the fractured vertebral body, and the sagittal plane kyphosis (SPK) which additionally describes an eventual destruction of the affected intervertebral disc. Compared with the preinjury status, the percentage of subjects who were able to do physical labor was reduced by half at follow-up, and four times as many patients had a permanent disability. Correspondingly, activity scores and Hannover spine scores declined significantly. After the initial surgical correction SI remained stable until follow-up, whereas SPK decreased again towards pre-operative values indicating a progressive deformity of the intervertebral disc space. Clinical results did not correlate with radiographic results, and neither the time until follow-up nor the type of fracture n or the use of transpedicular bone grafting affected clinical or radiographic results significantly. PMID- 12091035 TI - Zoophilia: a rare cause of traumatic injury to the rectum. PMID- 12091036 TI - Reliability of gastric tonometry after traumatic stomach herniation. PMID- 12091037 TI - Atypical traumatic sacrolisthesis following minimal trauma. PMID- 12091038 TI - Self-inflicted transcranial stab wound of the pons. PMID- 12091039 TI - Locked-in state due to vertebral artery thrombosis. PMID- 12091040 TI - Two in one: a variant of the Stener lesion. PMID- 12091043 TI - Prekallikrein deficiency: the characteristic normalization of the severely prolonged aPTT following increased preincubation time is due to autoactivation of factor XII. AB - Hereditary plasma prekallikrein (PK) deficiency was diagnosed in a 71-year-old man with an 8-year history of osteomyelofibrosis. PK deficiency was suspected in view of a severely prolonged activated partial thromboplastin time (aPTT) that nearly normalized following prolonged preincubation (10 min) of patient plasma with kaolin-inosithin reagent. Hereditary PK deficiency was demonstrated by very low PK values in the propositus (PK clotting activity 5%, PK amidolytic activity 5%, PK antigen 2% of normal plasma, respectively) and half normal PK values in his children. Normalization of a severely increased aPTT (>120 s) after prolonged preincubation with aPTT reagent occurred in plasma deficient in PK but not in plasma deficient in factor XII (FXII), high-molecular-weight kininogen (HK), factor XI (FXI), factor IX, factor VIII, Passovoy trait plasma or plasma containing lupus anticoagulant. Autoactivation of FXII in PK-deficient plasma in the presence of kaolin paralleled the normalization of aPTT. Addition of OT-2, a monoclonal antibody inhibiting activated FXII, prevented the normalization of aPTT. We conclude that the normalization of a severely prolonged aPTT upon increased preincubation time (PIT), characteristic of PK deficiency, is due to FXII autoactivation. PMID- 12091044 TI - Deficiency of von Willebrand factor-cleaving protease activity in the plasma of malignant patients. AB - von Willebrand factor (vWF) multimeric pattern and von Willebrand factor-cleaving protease activity (vWF-cp) were studied using plasmas from patients with advanced stage- and limited stage-malignant tumors. Deficiency of highly polymeric forms of vWF was observed in plasmas from 7 of 11 patients tested. vWF-cp activity was deficient in plasma samples of six patients with advanced stage-malignant tumors (ranging from 6% to 30% activity of normal plasma), whereas an essentially normal vWF-cp activity was observed in samples taken from patients with limited stage malignant tumors. Inhibitor of vWF-cp was not detected in any plasma samples tested. To further analyze the relevance of this enzymatic activity in metastatic diagnosis, a study of vWF-cp activity was conducted in 17 patients with colon cancer, and it was shown that deficiency of vWF-cp was associated with the progression of the disease. PMID- 12091045 TI - Low molecular weight heparin (Innohep) as thromboprophylaxis in outpatients with a plaster cast: a venografic controlled study. AB - INTRODUCTION: The aim of this study was to investigate the incidence of deep vein thrombosis (DVT) in patients immobilized in plaster cast and the possible efficacy of prophylaxis with low molecular weight heparin (LMWH). MATERIAL AND METHODS: The study was a randomized, assessor-blinded, open multicenter (three centers) study. All patients over 18 years of age with planned plaster cast on a lower extremity of at least 3 weeks were eligible for participation. Written informed consent was obtained from 300 patients and they were randomized to either 3.500 IU anti-Xa of tinzaparin (Innohep) subcutaneously once daily or no prophylaxis. On the day the cast was removed, ascending unilateral venography was performed. Two experienced radiologists, unaware of treatment, assessed the pictures independently. The radiologist had to obtain consensus as to whether DVT was present or not. RESULTS: 300 patients were included (148 in the treatment group and 152 in the control group). Ninety-five were subsequently withdrawn. DVT was diagnosed in 10/99 patients in the treatment group and in 18/106 patients in the control group. This difference is not significant (P=.15, chi(2) test) and the odds ratio was 0.55 (95% confidence interval=0.34-1.26). CONCLUSION: DVT in legs after plaster casting is a big problem, with an incidence of almost 20%. An effective prophylactic regime is required. Once-daily dose of 3.500 IU anti-Xa of tinzaparin was not sufficient. PMID- 12091046 TI - Retained fibrinolytic response and no coagulation activation after acute physical exercise in middle-aged women with previous myocardial infarction. AB - Sudden physical exertion is associated with an increased risk of acute myocardial infarction (MI) and sudden cardiac death. In addition, activation of the coagulation cascade and/or reduced fibrinolytic capacity after physical exercise has been reported in patients with cardiovascular disease. We investigated the haemostatic responses to an acute submaximal physical exercise in middle-aged women with a history of MI compared with healthy, age-matched controls. Resting plasma von Willebrand factor antigen (vWF Ag) and tissue plasminogen activator (tPA) antigen concentrations and plasminogen activator inhibitor-1 (PAI-1) activity were higher in the patients compared with control subjects. After 30 min of submaximal exercise on a bicycle ergometer, small, but still significant, increases in fibrinogen and vWF Ag concentrations were found in both groups. However, exercise did not induce thrombin generation and fibrin formation, as assessed by thrombin-antithrombin complex and fibrin D-dimer, in either group. Both tPA antigen concentration and activity increased and PAI-1 activity decreased significantly with exercise in both groups. Interestingly, the magnitude of changes in these latter variables did not differ between the groups (P=.99, P=.88 and P=.24, respectively). The present study demonstrates that some middle-aged women with previous MI have no signs of coagulation activation and retained fibrinolytic response after submaximal exercise. The clinical implication of these results might be that women with stable coronary heart disease can participate in rehabilitative exercise training without exhibiting a procoagulative state. PMID- 12091048 TI - Effects of coagulation Factor VII polymorphisms on the coronary artery disease in Japanese: Factor VII polymorphism and coronary disease. AB - We investigated the relationships among Factor VII coagulant activity (FVIIc), genetic polymorphisms of Factor VII (FVII) and coronary artery disease (CAD) in 380 unrelated Japanese individuals (mean 64 years) who underwent coronary angiography and whose cholesterol levels were within normal range. CAD subjects were defined as those in whom one of the three major coronary arteries showed >50% narrowing after nitroglycerin administration. FVIIc was measured and the following polymorphisms of FVII were determined: R353Q polymorphism (M1, M2 alleles), -323 0/10 bp polymorphism (0, 10 alleles), hypervariable region 4 of intron 7 (HVR4; H5, H6, H7 alleles). FVIIc was slightly lower in M1M2/M2M2 than M1M1 (89.5+/-8.9%, 93.4+/-17.8%). Those with M2 and/or 10 allele have less chance of developing CAD (M2: OR 0.36, 95% CI 0.18-0.69, 10: OR 0.50, 95% CI 0.26-0.97). However, both alleles did not associate with myocardial infarction (MI). HVR4 was unrelated with CAD, nor with MI. In conclusion, M2 and/or 10 allele has protective effects on the developing CAD in individuals with a normal cholesterol level. PMID- 12091047 TI - Thrombolytic therapy reduces red blood cell aggregation in plasma without affecting intrinsic aggregability. AB - Red blood cell (RBC) aggregation may contribute to occlusion of the coronary microcirculation during myocardial infarction. We studied the effect of thrombolytic therapy on RBC aggregation in patients with acute myocardial infarction (AMI). Compared with patients with myocardial infarction who did not receive thrombolytic therapy, those treated with systemic thrombolysis exhibited significantly reduced RBC aggregation, reduced plasma fibrinogen levels and increased plasma D-dimer levels. Using measurement of RBC aggregation in a standardized dextran-500 solution, reduction in RBC aggregation after thrombolysis was shown to be plasma dependent. Thrombolytic therapy had no direct effect on intrinsic RBC aggregability in patients with AMI. We conclude that thrombolytic therapy has rheologic consequences that may contribute to its overall efficacy. Inhibition of RBC aggregation by thrombolytic therapy may result from the degradation of fibrinogen, a key factor in the formation of RBC aggregates, and from the generation of fibrinogen degradation products capable of disaggregating RBCs. PMID- 12091049 TI - Influence of freeze-drying on the clotting properties of fibrinogen in plasma. AB - Freeze-dried plasma standards are often used to calibrate fibrinogen assays. Little is known, however, about the effect of freeze-drying on the clotting properties of fibrinogen. If these properties are altered, the use of freeze dried calibration standards might influence the results obtained when applying clotting assays to determine fibrinogen concentrations. In order to disclose any discrepancies in fibrinogen concentrations before and after freeze-drying, we determined the fibrinogen concentrations in citrated plasma samples using a total clottable protein method and a clotting-rate assay before and after freeze drying. When using the clotting-rate assay, significantly higher fibrinogen concentrations were found in fresh-frozen plasma samples compared to freeze-dried samples (P<.001). In freeze-dried plasma samples, the fibrinogen concentrations were significantly higher using the total clottable protein assay than the clotting-rate assay (P<.001). When measuring the fibrinogen concentrations in plasma samples with a wide range of fibrinogen concentrations using the routinely employed clotting-rate assay, significantly higher fibrinogen concentrations were found using the freeze-dried calibration plasma, than the fresh-frozen calibration plasma (P=.02). We conclude that the clotting rate of fibrinogen in citrated plasma is reduced following freeze-drying. When using freeze-dried calibration plasma in a clotting-rate assay, higher fibrinogen concentrations are obtained than by using fresh-frozen plasma. Denaturation of fibrinogen during the freeze-drying process, affecting its polymerization properties, may constitute the main contributor to the reduced clotting rate of freeze-dried plasma. PMID- 12091050 TI - Thrombin formation in platelet-rich plasma after oral methionine loading: preliminary report. PMID- 12091051 TI - There is no valid evidence presented as to an impaired endothelial NO system in the stroke-prone spontaneously hypertensive rats. AB - Platelet reactivity in stroke-prone spontaneously hypertensive rats (SHRSP) and normotensive Wistar-Kyoto rats (WKY) were compared. In vivo platelet reactivity was tested by the He-Ne laser-induced thrombosis model. The number of laser pulses needed to reach thrombotic occlusion of the targeted vessel was used as an index of thrombogenicity. SHRSP rats needed significantly less number of irradiation to reach occlusion than WKY rats (SHRSP vs. WKY, 5.1+/-0.3 vs. 8.1+/ 0.6), indicating enhanced thrombotic response in SHRSP rats. Further, acetylcholine administration significantly increased the number of laser pulses until occlusion in WKY but not in SHRSP rats. This suggests an impaired thrombotic reaction in acetylcholine-treated WKY but not in SHRSP rats. Platelet reactivity in vitro was measured in native blood by a shear-induced haemostasis test (haemostatometry). Indexes of this test (H1/H2), which inversely correlated with platelet reactivity, were significantly greater in SHRSP than in WKY rats (SHRSP vs. WKY, H1: 1815+/-192 vs. 763+/-75; H2: 7547+/-723 vs. 3536+/-264). This suggests reduced platelet reactivity in SHRSP compared with WKY rats. Thus, the present findings show increased thrombotic tendency in SHRSP rats in vivo despite reduced platelet reactivity in vitro. To explain this contradiction, we suggest that an increased in vivo thrombotic tendency may be due to impaired nitric oxide (NO) release from endothelial cells in SHRSP rats, and that a reduced platelet reactivity in vitro may be due to an adaptation of SHRSP rats to survive at extremely high blood pressure. PMID- 12091052 TI - Combined Ala601-Thr-type dysplasminogenaemia and antiphospholipid antibody syndrome in a patient with recurrent thrombosis. AB - A 44-year-old man was admitted to our hospital because of recurrent thrombosis. Laboratory findings showed that he had antiphospholipid antibody syndrome (APS) associated with systemic lupus erythematosus. Plasminogen activity was reduced (21%), but its antigenic level was rather elevated (16.4 mg/dl). The discrepancy of plasminogen activity and antigen levels was also found in his three daughters. Gene analysis revealed a homozygous missense mutation (Ala601-->Thr) at exon 15 of the plasminogen gene in the patient and a heterozygous mutation in his three daughters, suggesting that the patient has dysplasminogenaemia, which was reported as "plasminogen Tochigi." Although it still remains controversial whether both dysplasminogenaemia and plasminogen deficiency are relevant independent thrombotic risk factors, the combination of this deficiency and APS should be considered as a strong predisposition to thrombosis in this patient. PMID- 12091053 TI - Analysis of the -1185A/G von Willebrand factor (VWF) gene polymorphism in two Brazilian ethnic groups and its effect on the plasma VWF levels. AB - Both genetic and environmental factors influence the variation in von Willebrand factor (VWF) levels between individuals, the main genetic variable known to be involved in differences in VWF levels being the ABO blood group. The -1185A/G polymorphism in the 5'-regulatory region of VWF gene has been associated with plasma VWF levels in a normal population. The objective of our study was to examine the relationship between the -1185A/G polymorphism and plasma VWF levels in a total of 420 individuals from two Brazilian ethnic groups. The -1185A/G genotypes were identified using polymerase chain reaction (PCR) amplification of the 864-bp VWF promoter region followed by AccII restriction digestion. Allele and genotype frequencies were significantly different between Afro-Brazilians and Euro-Brazilians. The -1185A allele frequency was 62% in Afro-Brazilians and 41% in Euro-Brazilians. Although Afro-Brazilians had higher plasma VWF levels than Euro-Brazilians, the ethnic differences were not significant. No association was observed between -1185A/G genotypes and VWF plasma levels in either ethnic groups. The present data indicate that there are no ethnic differences in VWF levels. The -1185A/G polymorphism showed significantly different frequencies between Afro-Brazilians and Euro-Brazilians, but in our study, it did not appear to play a role in the determination of VWF levels. PMID- 12091054 TI - Reductions in platelet contractile force correlate with duration of cardiopulmonary bypass and blood loss in patients undergoing cardiac surgery. AB - Blood loss secondary to platelet dysfunction is known to be increased when the duration of cardiopulmonary bypass (CPB) is prolonged. The ability to correlate alterations in platelet function with the duration of bypass and early postoperative blood loss, however, has remained elusive. Platelet contractile force, a novel measure of platelet-mediated clot retraction, is known to be reduced following cardiac surgery and blockade of platelet adhesion receptors. The aim of this study was to determine if alterations in platelet contractile force (measured using whole blood) correlated with the duration of CPB and early postoperative blood loss. Thirty patients were entered into a study designed to measure platelet function before, during, and after CPB. Platelet aggregometry and surface expression of CD42b and CD61 were also measured (using whole blood) in a subset of subjects (n=10) to further characterize the intrinsic structural and functional defects induced by CPB. Reductions in platelet contractile force had a significant correlation with duration of CPB (r=0.564; P=0.002) and early blood loss (r=0.545; P=0.003). Although decreases in platelet contractile force and aggregation both correlated with CPB time in the smaller subset of patients tested, only platelet contractile force correlated with decreases in CD42b, CD61 and blood loss. The results of this study suggest that prolongation of CPB is related to increasing degrees of platelet dysfunction and that reductions in platelet contractile force are related to decreases in platelet adhesion receptors and early postoperative blood loss. PMID- 12091056 TI - Antithrombin and first complement protein recognize the same active heparin fraction. AB - Antithrombin (AT) high affinity of unfractionated heparin (UFH) resides in a specific pentasaccharide sequence. Heparin also regulates complement activity on the classical and the alternative pathways. Most experimental pieces of evidence accumulated show that these important activities reside in different segments of the heparin molecule. We demonstrated in previous papers that a low ionic strength and the presence of calcium ions are essential to detect specific interactions between glycosaminoglycans and proteins. Then these very strict conditions were used, and we demonstrated that the first protein complex of the human complement cascade recognizes in the UFH a fraction with very high anticoagulant activity. After isolation from the precipitate of the interaction, this fraction of heparin also contained the pentasaccharide sequence responsible for the great affinity with AT: in fact, it was strongly bound to a resin of AT agarose, and to detach it, an ionic strength of 0.6 M sodium chloride was required. In this way, the heparin regions responsible for the anticoagulant activity and also for the effects over the complement system were identified on the same short segment of the heparin molecule, which includes the active fraction of the glycosaminoglycan. The differences with early results could be explained by our experimental conditions of low ionic strength and the presence of calcium ions used for the interaction of the protein and the glycosaminoglycan. PMID- 12091055 TI - L-158,809 and (D-Ala(7))-angiotensin I/II (1-7) decrease PAI-1 release from human umbilical vein endothelial cells. AB - The endothelium is a major source of plasminogen activator inhibitor-1 (PAI-1), which plays a critical role in the regulation of fibrinolysis. There are many reports on the increase in the expression of PAI-1 by angiotensin II (Ang II). In the present study, we investigated the effects of angiotensin-related substances on the release of PAI-1 from human umbilical vein endothelial cells (HUVECs). Ang II increased PAI-1 and tissue plasminogen activator (t-PA) release, while its metabolite angiotensin-(1-7) (Ang-(1-7)) amino acid fragment decreased them. Angiotensin Type 1 (AT1) receptor antagonist, L-158,809 (L-1), and Ang-(1-7) receptor antagonist, (D-Ala(7))-angiotensin I/II (1-7) (D-Ala), decreased PAI-1 and t-PA release; angiotensin Type 2 (AT2) antagonist, PD123,319 (PD), however, did not have any effects on the release of PAI-1 and t-PA. The addition of the equal concentration or 10-times-higher concentration of L-1 to Ang II did not change PAI-1 release compared to that by Ang II. Although Ang-(1-7) and L-1 decreased PAI-1 release, there were no additional effects on the decrease of the amounts of PAI-1 by the mixture of Ang-(1-7) and the equal concentration or 10 times-higher concentration of L-1 compared to those by Ang-(1-7). The equal concentration of D-Ala to Ang II did not change the amounts of PAI-1, but the addition of the 10-times-higher concentration of D-Ala to Ang II resulted in significant decrease of the amounts of PAI-1 compared to those by Ang II. The addition of equal concentration or 10-times-higher concentration of D-Ala to Ang (1-7) showed the significant decrease of the amounts of PAI-1 compared to those by Ang-(1-7). In conclusion, L-158,809 and (D-Ala(7))-angiotensin I/III (1-7) may be used as profibrinolytic drugs. PMID- 12091057 TI - Prothrombin time quality assessment scheme in the Basque Country. AB - The Basque Country Quality Assessment Scheme on Oral Anticoagulant Treatment (PEEC-CAV) was first launched in 1984. This project combined prothrombin time (PT) quality controls and a new in-house standard reagent called Thromboplastin Bilbao (TBi). Originally, this reagent was of human origin, but in 1994, it was replaced by one derived from rabbit brain following WHO recommendations. Nine hospitals in the area collaborated in carrying out quality controls (a) to assess different systems of PT test performance in the Basque Country and (b) to evaluate TBi response in these controls. Evaluation over the period 1994-1999 yielded an INR coefficient of variation (CV) for the plasmas used in these controls of 8.29+/-4.18% using the laboratories' routine system (reagent and coagulometer), and 9.7+/-2.70% using TBi and the manual technique. These results were similar to those obtained in the CV of PT ratio with human brain thromboplastin from 1984 to 1988 (9.26+/-2.84%). PMID- 12091058 TI - Editorial comment on "In vitro toxicity of ET-743 and Aplidine, two marine derived antineoplastics, on human bone marrow haematopoietic progenitors: comparison with the clinical results" by Albella and colleagues. PMID- 12091059 TI - Diagnostic advances and new trends for the treatment of primary central nervous system lymphoma. AB - Primary central nervous system lymphoma (PCNSL) is a rare non-Hodgkin's lymphoma arising in the brain. Recent increase in its incidence has been noted both in immunocompetent individuals and patients with immunodeficiency. This review will focus on the epidemiology, pathogenesis, diagnosis and treatment of this aggressive extranodal lymphoma in immunocompetent patients. Stereotactic biopsy is usually required for diagnosis, while molecular biology and/or cytofluorimetric analysis may confirm the presence of clonal proliferation in the cerebrospinal fluid (CSF). Methotrexate-based chemotherapy plus whole-brain radiotherapy are the standard treatment for PCNSL and achieve a high rate of complete remissions (CR), but long-term neurotoxicity may heavily compromise the patient's quality of life. The metabolic rate of controversial gadolinium enhancing lesions on magnetic resonance (MR) scans may be assessed with positron emission tomography (PET), which discriminates radiation necrosis from true recurrence. Withholding radiotherapy in patients achieving CR after first-line chemotherapy is a new and interesting treatment option, while the role of high dose chemotherapy with stem cell rescue is still uncertain. PMID- 12091060 TI - Mind and cancer. do psychological factors cause cancer? AB - We have reviewed the evidence for an association between major life events, depression and personality factors and the risk for cancer. We identified and included only those prospective or retrospective studies in which the psychological variable was collected independently of the outcome. The evidence failed to support the hypothesis that major life events are a risk factor for cancer. The evidence was inconsistent for both depression and personality factors. Chance, bias or confounding may explain this result, as many of the studies had methodological weaknesses. The generally weak associations found, the inconsistency of the results, the unresolved underlying biological mechanism and equivocal findings of dose-response relationships prevent a conclusion that psychological factors are established risk factors. However, certain intriguing findings warrant further studies, which must, however, be well conducted and large and include detailed information on confounders. PMID- 12091061 TI - Alveolar soft part sarcoma. a report of 15 cases. AB - The aim of this study was to evaluate the presentation, course and treatment outcome of 15 patients with this rare type of sarcoma. The files of the patients were retrospectively analysed. Overall survival was calculated according to the Kaplan-Meier method. There were 15 patients, 8 male and 7 female. The mean age at diagnosis was 29 years for men and 24 years for women. The median survival was 48 months with an overall 5-year survival of 38%. 5 patients had haematogenic metastases at the time of diagnosis. For the remaining 10 patients with localised disease, the median survival was 48 months and the 5-year survival 48%. The median disease-free survival for these patients was 12 months with a 5-year disease free survival of 40%. After the occurrence of haematogenic metastases, patients survived a median period of 8 months (range 0-45 months). 5 patients are still free of disease after a median period of 234 months (12-295 months). Alveolar soft part sarcoma is found especially in young adults. When diagnosed, it is often metastasised with a poor prognosis. However, when radically resected, long-term survival is possible. PMID- 12091062 TI - Factors influencing the prognostic role of oestrogen and progesterone receptor levels in breast cancer--results of the analysis of 670 patients with 11 years of follow-up. AB - In the last two decades, the prognostic role of the steroid hormone receptors has been the subject of a myriad of publications. Nevertheless, its relevance after long-term follow-up is still not clear. The confusion about the prognostic value is mainly due to the difficulty in comparing analyses. Despite different study designs and statistical approaches, oestrogen (ER) and progesterone (PR) receptors are widely accepted as prognostic factors. Data from 670 breast cancer patients with a median follow-up of 11.4 years were analysed retrospectively. ER and PR were measured by the dextran-coated charcoal (DCC) assay. To investigate the time dependence of the prognostic relevance of ER and PR, separate analyses were done for follow-up shorter and longer than 5 years. Special focus was directed at patients < or =50 and >50 years, node-negative women, in particular those without adjuvant therapy. Univariate and multivariate analyses were performed. In univariate analysis, ER and PR were associated with a significantly longer overall survival at the cut-off levels 10, 20 or 100 fmol/mg protein. The significant survival benefit occurred in the first 5 years of follow-up and remained unchanged in the following period. In the multivariate analyses, only the PR was of significant prognostic value (for PR> or =20 fmol/mg P=0.036, for PR> or =100 P=0.01, Cox analysis). In patients younger than 51 years, only PR was an independent prognosticator at the cut-off level of 100 fmol/mg protein, while in patients >50 years both hormone receptors were not significant. In N0 patients, only the PR reached long-term prognostic independence at a cut-off point of > or =100 fmol/mg (P=0.018). In addition, in the group of node-negative women < or =50 years without adjuvant therapy the PR level reached prognostic significance. The hormone receptor status was a prognostic factor only during the first 5 years of follow-up. Our data suggest that age, lymph node status, length of follow-up and probably the ER/PR assay are important for the evaluation of ER and PR as prognostic variables. In most analyses, PR appeared to be superior to ER in predicting the prognosis of primary breast cancer patients. PMID- 12091063 TI - ErbB-4 expression in limb soft-tissue sarcoma: correlation with the results of neoadjuvant chemotherapy. AB - ErbB-4 is a recently described growth factor receptor. Relatively little is known about its expression in human tumours. In this study, we assessed the possible role of erbB-4 as a tissue marker for soft-tissue sarcomas (STS) and its correlation with the response to chemotherapy. The histological specimen of 29 patients with STS of a limb who had received preoperative doxorubicin (ADR)-based chemotherapy were studied for the degree of necrosis and the expression of erbB-4 (by an avidin-biotin-peroxidase technique). ErbB-4 expression in the preoperative tissue samples was compared with the expression in the postchemotherapy resected tumour. The true objective response rate to preoperative chemotherapy was 34%. Wide resection of the tumour was done in 12 patients, marginal in 14, amputation in 2 and no surgery in 1. The tumour necrosis was above 90% in 9 patients, 60-90% in 12, and less than 60% in 7 patients. An increase in erbB-4 expression was more common in cases with no response to chemotherapy, while no change or a decrease in erbB-4 was more common in responsive tumours (P=0.004). No correlation could be found between the degree of necrosis or the chemotherapeutic regimen and the change in expression of erbB-4. The median disease-free survival (DFS) was longer for patients with a decrease or no change in expression of erbB-4 than for patients with increased expression. It is believed that postchemotherapy new expression or no downregulation of the erbB-4 molecule represents tumour aggressiveness and increased capability of growth and spread. PMID- 12091064 TI - High expression of DNA topoisomerase IIalpha and Ki-67 antigen is associated with prolonged survival in glioblastoma patients. AB - Assessment of tumour cell proliferation in glioblastoma (GB) has been a topic of considerable research interest over the past decade. However, the correlation of tumour proliferation and patient outcome has yielded controversial results. In this study, we examined immunohistochemically, using paraffin-embedded tissue, the expression of the proliferation-related markers DNA topoisomerase IIalpha (TIIalpha) and Ki-67 antigen in a cohort of 114 GB patients treated consecutively with surgery and radiochemotherapy, and correlated the expression with patient outcome. The TIIalpha labelling index (LI) ranged between 5.2 and 87.2% (median: 25.6%). Survival analysis disclosed an association between high TIIalpha expression levels and prolonged survival (P=0.040, log-rank test). TIIalpha expression correlates closely with Ki-67 labelling index (R=0.927, P<0.001), which itself is predictive of patient survival (P=0.044). However, in multivariate analysis, only the Karnofsky performance status remained predictive of patient survival. We conclude that high expression of TIIalpha and Ki-67 appears to be associated with a prolonged survival in our cohort of GB patients. PMID- 12091065 TI - Multicentre phase II and pharmacokinetic study of RFS2000 (9-nitro-camptothecin) administered orally 5 days a week in patients with glioblastoma multiforme. AB - A phase II trial was instigated to investigate the antitumour activity, the safety and the pharmacokinetic parameters of RFS2000, a recently identified oral topoisomerase I inhibitor, given once daily (1.5 mg/m(2)/day) as first-line chemotherapy treatment for patients with advanced glioblastoma multiforme (GBM). Between 9 March and 15 September 2000, 17 patients were entered onto the trial. 15 patients were considered eligible. A total of 49 cycles (range 1-8) were administered. Grade 3-4 toxicity was observed in 5 patients. Neutropenia and thrombocytopenia were common toxicities. Pharmacokinetic analysis showed that 9 nitro camptothecin (9-NC) could be detected in the plasma and is progressively converted into 9-amino-camptothecin (9-AC). The response rate was poor, with 5 patients experiencing tumour stabilisation and 10 progressing. Thus, the results do not support the further evaluation of RFS2000 as a single agent in patients with recurrent GBM. PMID- 12091066 TI - Baseline quality of life predicts survival in patients with advanced colorectal cancer. AB - The aim of this study was to investigate the influence of baseline quality of life (QoL) on survival in patients with advanced colorectal cancer. From 1992 to 1998, four randomised clinical trials in advanced colorectal cancer were conducted at this institution. The European Organization for Research and Treatment of Cancer-Quality of Life Core 30 (EORTC-QLQ-C30) questionnaire was completed prior to the commencement of chemotherapy. Analyses were performed on median-dichotomised baseline Quality of Life (QoL) and clinical prognostic factors. Baseline QoL questionnaires were completed by 501 patients. One-year survival was 38.3 and 72.5% (P<0.0001) for patients with global QoL scores below and above the median (67), respectively. Other than cognitive functioning, fatigue, appetite, constipation, diarrhoea and financial domains, all QoL scales were significant independent predictors of survival (P<0.035). In the final model, the global QoL score remained highly significant as an independent predictor of survival (P<0.0001). Baseline QoL is a strong independent predictor of survival in patients with advanced colorectal cancer. Measurements should be routinely recorded in clinical trials to stratify cohorts and aid in trial comparison. PMID- 12091067 TI - A randomised controlled study on the use of anti-inflammatory drugs in patients with cancer pain on morphine therapy: effects on dose-escalation and a pharmacoeconomic analysis. AB - The role of non-steroidal anti-inflammatory drugs (NSAIDs) in cancer pain has been well established in the treatment of mild pain and in association with opioids in the treatment of moderate to severe pain. The aim of this study was to verify the effects of NSAIDs on morphine escalation in advanced cancer patients with pain followed-up at home and to assess the pharmacoeconomic implications. A prospective randomised controlled study was carried out in 156 consecutive advanced cancer patients with pain followed-up at home in the period December 1999-December 2000. In this group of patients, 47 were selected with pain progression after 1 week of opioid stabilisation. Patients were randomly assigned to one of two groups: group 'O' patients were treated with continuing opioid escalation according to their clinical needs; group 'OK' received ketorolac 60 mg/daily orally (p.o.) in three doses and then continued opioid escalation according to their clinical situation. Performance status, doses of morphine before and after starting treatment, mean weekly pain intensity (assessed by means of a numerical scale from 0 to 10), mean weekly symptoms intensity, adverse effects and pain mechanisms were recorded. Moreover, drug costs per day in both groups were calculated. Patients who received ketorolac in addition to morphine showed a better analgesia after a week in comparison to the group treated with morphine only (P=0.005). Thereafter, morphine escalation was slower and the maximum morphine dose was lower in the group treated with ketorolac. The incidence and the severity of gastric discomfort was more evident in patients treated with ketorolac, while constipation was significantly increased in patients who received morphine only. Drug costs per day were similar in both groups; statistical differences were observed in patients who started on lower morphine doses (<100 mg/daily) in the two groups (4.3 in the ketorolac-morphine group versus 3.4 in the morphine group; P=0.012). The use of NSAIDs reduces the need for an opioid dose escalation or allows the use of lower doses. Their use is associated with a more intense gastric discomfort, but results in less opioid related constipation. The eventual additive cost for NSAIDs therapy is negligible, especially in patients taking high doses of morphine. PMID- 12091068 TI - Emergencies and their management. PMID- 12091070 TI - An evaluation of chest X-ray screening for lung cancer in gunma prefecture, Japan: a population-based case-control study. AB - In order to evaluate the efficacy of annual chest X-ray screening for lung cancer, a case-control study was conducted in Gunma Prefecture, Japan. Population based annual lung cancer screening programmes have been conducted by the local government in Gunma Prefecture since the mid-1970s. A total of 121 case subjects, including 91 high-risk males and 30 non-high-risk females between the ages of 40 and 79 years who died of lung cancer from 1992 to 1997 were evaluated. A total of 536 controls (3-5 controls for each case) were matched to case subjects by gender, year of birth, address and smoking habits. Controls were selected from screening programme lists provided by the local governments. All case subjects were also included on these lists. The smoking-adjusted odds ratio (OR) of lung cancer death for those subjects screened within 12 months prior to diagnosis versus those not screened was 0.68 (95% confidence interval (CI): 0.44-1.05; P=0.084). When the analysis was conducted without matching case and control subjects by smoking habits, the OR was 0.79 (95% CI: 0.53-1.18). When stratified by histological type, the OR was 0.62 (95% CI: 0.31-1.24) for adenocarcinoma, and 1.01 (95% CI: 0.44-2.31) for squamous cell carcinoma. The results of this study suggest 20-30% of deaths attributable to lung cancer, especially adenocarcinoma, might be prevented by annual chest X-rays. PMID- 12091071 TI - Survival probabilities and hazard functions of malignant melanoma in Germany 1972 1996, an analysis of 10433 patients. Evolution of gender differences and malignancy. AB - The evaluation of the impact of prevention activities on the course of survival in conjunction with the individual hazard rate of dying is described using data from a follow-up study of 10433 melanoma patients during three observation periods (1972-1980, 1981-1988, 1989-1996). Kaplan-Meier survival curves combined with hazard functions were calculated. At all observation periods, survival of men was lower compared with women and their maximum dying risk was earlier (70 versus 100 months after removal of the primary tumour). In 1989-1996, differences in the survival rates were approximately halved compared with those for 1972-1980 or 1981-1988, respectively. This improvement was predominantly seen in young men. There was a lower survival rate of men compared with women with identical thickness categories. The maximum dying risk for those men with tumours >4 mm peaked at approximately 60 months, the other thickness categories showing a lower and later maximum; in women, the maximum dying risk for tumours >4 mm was also seen at approximately 60 months, but less pronounced. Over time, the influence of Breslow thickness on the survival rates remained constant in women; in men, with the exception of thick tumours, there was a trend towards a better survival. Melanoma awareness campaigns conducted in Germany since the late 1980s have resulted in a trend towards a remarkable increase of thin tumours in recent years, whereas the number of new cases with thick tumours has remained constant. PMID- 12091072 TI - In vitro toxicity of ET-743 and aplidine, two marine-derived antineoplastics, on human bone marrow haematopoietic progenitors. comparison with the clinical results. AB - Ecteinascidine-743 (ET-743) and aplidine are two marine-derived antineoplastics currently in phase II development. With the aim of evaluating whether in vitro haematopoietic studies can predict the toxicity of these two drugs in patients, human bone marrow (BM) samples were incubated with these drugs under conditions which mimicked the administration exposures used in the clinics. As it was observed in different cancer cell lines, ET-743 was more toxic on an equimolar basis in human hematopoietic progenitors (inhibitory concentration reducing the viability to 50% after 24 h exposures; IC50(24h): 10-50 nM) compared with doxorubicin (IC50(24h) values: 280-460 nM), used as a control anticancer drug. In contrast to the high haematotoxic effects observed for ET-743, similar IC values were obtained for aplidine (IC50(24h): 150-530 nM) compared with doxorubicin. For both ET-743 and aplidine, the megakaryocytic progenitor was the most sensitive, compared with the other haematopoietic progenitors (IC50 values were 3- to 5-fold lower in the CFU-Megs compared with the CFU-GMs). The observation that the Cmax observed in patients treated with the aplidine maximum tolerated dose (MTD) (7.1 nM) was 21-75 fold lower than the IC50(24h) value observed for the different haematopoietic progenitors is highly consistent with the lack of haematotoxicity observed in patients treated with this drug. In the case of ET-743, differences between the Cmax value corresponding to the MTD (2.6 nM) and the in vitro IC50 values corresponding to the different progenitors were much lower (4-19-fold), also consistent with the haematotoxicity that was observed in patients treated at recommended doses (RDs) and MTDs. Although CFU-Megs were more sensitive than CFU GM progenitors to ET-743 in vitro, clinical data showed that neutropenic events were more frequent than thrombocytopenic episodes. Aiming to further improve the predictive value of in vitro IC values corresponding to the different haematopoietic progenitors, additional refinement parameters derived from pharmacokinetic and animal studies are proposed. PMID- 12091073 TI - The cellular pharmacology of oxaliplatin resistance. AB - Oxaliplatin is a third generation platinum compound that differs from cisplatin and carboplatin in having a broader spectrum of antitumour activity. Molecular studies suggest that oxaliplatin adducts are recognised and processed differently than those produced by the earlier generation Pt-containing drugs. We report here studies on the kinetics of the development of oxaliplatin resistance, and the changes in the cellular pharmacology of oxaliplatin that accompany the emergence of the resistant phenotype in five parental human tumour cell lines and their sub lines selected for acquired oxaliplatin resistance in vitro. During selection, resistance did not substantially increase until after at least six cycles of oxaliplatin treatment. Oxaliplatin demonstrated schedule-dependency with a 1-h exposure being substantially less cytotoxic than a continuous exposure. Whole cell uptake was linear with concentration, but uptake in the resistant cells averaged only 27+/-10 S.D.% of that in the sensitive cells. Pt accumulation in DNA was markedly reduced in four of the five resistant cell lines, but this did not correlate with either IC(50) or total cellular accumulation. Four of the five resistant sub-lines also demonstrated increased tolerance to adducts in DNA that ranged from 3.1 to 7.6-fold. We conclude that development of acquired resistance to oxaliplatin is accompanied by independent defects in both whole cell uptake and in adduct formation. PMID- 12091075 TI - Introduction to the virological diagnosis of the most frequent infections of the central nervous system. PMID- 12091074 TI - Vascular endothelial growth factor-C expression predicts lymph node metastasis of human gastric carcinomas invading the submucosa. AB - We examined the relationship between vascular endothelial growth factor (VEGF)-C expression and lymph node metastases in gastric carcinomas invading the submucosa. Of the six human gastric carcinoma cell lines, two constitutively expressed VEGF-C mRNA. In three of 12 gastric biopsy specimens (25%), VEGF-C mRNA was detected in tumour tissues, but not in corresponding normal mucosa by reverse transcriptase-polymerase chain reaction (RT-PCR). Of the 139 resected gastric carcinomas, 44 (32%) showed intense cytoplasmic VEGF-C immunoreactivity in many cancer cells at the invading edge. VEGF-C immunoreactivity was associated with greater depth of tumour invasion, lymphatic invasion and lymph node metastases. In addition, vessel count was also significantly higher in the VEGF-C immunoreactive tumours than in other tumours. These results suggest that VEGF-C may be involved in the progression of human gastric carcinoma, particularly via lymphangiogenesis. VEGF-C expression at the invading edge of a gastric carcinoma may be a sensitive marker for metastasis to the lymph nodes. PMID- 12091076 TI - Use of PCR for the diagnosis of herpesvirus infections of the central nervous system. AB - Polymerase chain reaction (PCR) analysis of cerebrospinal fluid (CSF) has revolutionized the diagnosis of nervous system viral infections, particularly those caused by human herpesviruses (HHV). The PCR technique allows the detection of minute quantities of DNA or RNA in body fluids and tissues. Both fresh-frozen and formalin-fixed tissues may be utilized for PCR assays, with the latter making archival studies possible. CSF PCR has now replaced brain biopsy as the gold standard for the diagnosis of herpes simplex virus (HSV) encephalitis. PCR analysis of both CSF and nervous system tissues has also broadened our understanding of the spectrum of disease caused by HSV-1 and -2, cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella zoster virus (VZV) and HHV-6. PCR results obtained from tissue specimens must be interpreted cautiously, since this highly sensitive technique may detect portions of viral genomic material that may be present even in the absence of active viral infection. Tissue PCR results in particular must be corroborated with clinical and neuropathologic evidence of central nervous system (CNS) infection. In several neurological diseases, negative PCR results have provided evidence against a role for herpesviruses as the causative agents. This review summarizes the role of CSF PCR in the diagnosis and therapeutic management of herpesvirus infections of the nervous system, particularly those caused by HSV and VZV. PMID- 12091077 TI - Comparison of mixed cell culture containing genetically engineered BGMK and CaCo 2 cells (Super E-Mix) with RT-PCR and conventional cell culture for the diagnosis of enterovirus meningitis. AB - BACKGROUND: Enteroviral meningitis has traditionally been diagnosed by cell culture. More recently, molecular techniques and shell vials with a mixture of human colon carcinoma and genetically engineered buffalo green monkey kidney cells (BGMK cells) (Super E-Mix) have been described. OBJECTIVE: We compared the results of this new cell culture technique with two reverse transcriptase polymerase chain reaction (RT-PCR) techniques and conventional cell culture to assess the accuracy of these various methods. STUDY DESIGN: 2 ml of cerebrospinal fluid (CSF) was obtained from 72 patients with suspected viral meningitis. 600 microl was used for conventional cell culture and 200 microl was inoculated to each of two Super E-Mix vials. One vial was incubated for 24 h, then stained with pan-enterovirus antibody, and the other was examined for CPE daily for 5 days and stained when positive or at the end of incubation. The final aliquot (500 microl) was centrifuged at 25,000g, and the pellet in 140 microl of supernatant was used for ribonucleic acid (RNA) extraction and tested by commercial single-step (Chemicon, Temecula, CA) and two-step (Argene, Varilhes, France) RT-PCR methods. RESULTS: Conventional culture was 51% sensitive and 100% specific. The Super E Mix was slightly higher (sensitivity 76%, specificity 100%). The Chemicon and Argene PCR methods had a sensitivity of 93% and 88% and specificity of 90% and 97%, respectively. CONCLUSION: The Super E-Mix procedure had greater sensitivity than conventional cell culture, but RT-PCR was the most sensitive technique with this type of specimen. The RT-PCR methods performed equally. PMID- 12091078 TI - The impact of an enteroviral RT-PCR assay on the diagnosis of aseptic meningitis and patient management. AB - BACKGROUND: Enterovirus (EV) is a major cause of aseptic meningitis and non specific febrile illness in children. Since the majority of patients are hospitalized for possible bacterial infection, a rapid test for the diagnosis of enteroviral meningitis (EVM) may reduce hospitalizations and unnecessary treatments. OBJECTIVE: To review the impact of an EV reverse transcriptase polymerase chain reaction (RT-PCR) assay for the diagnosis of EVM on patient management. STUDY DESIGN: CSF from 1056 patients admitted to the hospital between 1998 and 2001 was tested using EV RT-PCR. The results were correlated with CSF counts, diagnosis, test turnaround time (TAT) and length of hospital stay (LOS). RESULTS: EV RT-PCR was positive for 113 patients (11%). Of these cases, 92% occurred during the summer months and 77% were in children <19 years of age. Children <3 years old with EVM frequently had non-specific clinical findings and lacked pleocytosis. There was a significant correlation between decreasing LOS and TAT (r(2)=0.97, P<0.001). CONCLUSION: RT-PCR testing for EVM is an important tool to aid in the diagnosis of children with non-specific febrile illness. This test impacted patient management as measured by shortened patient stays, which should translate into significant health care savings. PMID- 12091079 TI - Application of polymerase chain reaction and subsequent phylogenetic analysis to the diagnosis of enteroviral infection in the central nervous system. AB - BACKGROUND: Enteroviral infections of the central nervous system (CNS) are often difficult to diagnose, even if consistent conventional laboratory methodologies are used. OBJECTIVES: To clarify the efficiency of two polymerase chain reaction (PCR) methods for the sensitive detection of enteroviruses and for the identification of enteroviral genotypes based on phylogenetic analysis of the amplified genome sequences, and to facilitate the diagnosis of enteroviral infection in CNS. STUDY DESIGN: Cerebrospinal fluid (CSF), throat swab, rectal swab, and/or serum samples were collected from 171 patients with aseptic meningitis and 67 patients with febrile seizures. The samples were tested for the presence of enteroviruses by cell culture and PCR methods for the detection and identification of enteroviruses. RESULTS: In 111 (64.9%) of 171 patients with aseptic meningitis, enteroviruses were isolated by cell cultures from any site. In 143 (83.6%) patients, including 110 of 111 patients with aseptic meningitis, the enteroviral genome was detected in CSF by PCR. No enterovirus was isolated from any site for the 67 patients with febrile seizures. PCR detected the enteroviral genome in CSF samples from 13 (61.9%) of 21 patients who developed febrile seizures in the summer (June-August). Phylogenetic analysis of amplified genome sequences showed that the major pathogens of febrile seizures in summer were group A coxsackieviruses, which are usually difficult to isolate by cell culture. CONCLUSION: PCR methods for the detection and identification of enteroviruses were useful for the diagnosis of enteroviral infection in CNS. PMID- 12091080 TI - Evaluation of a rapid real-time RT-PCR assay for detection of enterovirus RNA in cerebrospinal fluid specimens. AB - BACKGROUND: We previously described the characteristics of a single-tube real time enterovirus reverse transcriptase polymerase chain reaction (RT-PCR) assay based on a fluorogenic probe and primers directed to highly conserved sequences in the 5'-untranslated region (UTR) of the enterovirus genome. OBJECTIVES: To evaluate the performance of the assay on a larger number of cerebrospinal fluid (CSF) specimens from patients suspected of having viral meningitis. STUDY DESIGN: Real-time enterovirus RT-PCR and viral culture were performed on CSF specimens received from March 2000 to November 2001. Patient records were retrospectively reviewed for final clinical diagnosis. RESULTS: From the 186 CSF specimens tested, culture was positive for enterovirus in 31 cases, whereas real-time RT PCR detected enterovirus RNA in 45 CSF specimens. The sensitivity of real-time RT PCR in relation to the clinical diagnosis of viral meningitis was 72.6%, whereas the sensitivity of viral culture reached only 57.4%. Enterovirus RNA was also found in a number of specimens with low leukocyte counts. CONCLUSIONS: We confirm that the real-time enterovirus RT-PCR assay for CSF specimens is significantly more sensitive than viral culture. PMID- 12091081 TI - Laboratory diagnosis of central nervous system infections caused by herpesviruses. AB - BACKGROUND: Herpesviruses may be associated with various types of central nervous system (CNS) infections. Herpes simplex encephalitis (HSE) has to be considered one of the most severe diseases. As effective antiviral drugs are available, rapid and reliable diagnosis has become important. OBJECTIVES: To describe polymerase chain reaction (PCR) and serological methods for the detection of herpesvirus-induced CNS infections by the example of HSE. STUDY DESIGN: 620 cerebrospinal fluid (CSF) and 2400 serum samples from 2700 selected hospitalized patients with clinical suspicion of encephalitis were tested for herpes simplex virus (HSV) as well as varicella-zoster virus (VZV) DNA and HSV-specific antibodies, respectively. RESULTS: HSV-1 DNA could be detected in eight and HSV-2 in three patients with focal encephalitis. In addition, HSV-2 DNA was found in two newborns with encephalitis and two adults suffered from transverse lumbar myelitis. One VZV DNA-positive patient had developed herpes zoster accompanied by meningoencephalitis, and in the other an encephalitis without cutaneous rash was diagnosed. Intrathecal antibody synthesis could be measured when CSF was cleared from viral DNA. CONCLUSIONS: The detection of viral DNA by PCR technique has become the "gold standard" method for laboratory diagnosis of herpesvirus infections of CNS. Serodiagnosis may be useful to confirm the diagnosis retrospectively. PMID- 12091082 TI - Detection of herpesvirus infection of the CNS: the experience of hospital Geral de Santo Antonio. AB - BACKGROUND: PCR detection of CSF Herpes virus DNA is an important tool in the diagnosis of CNS infections. Use of this test has been shown to have an impact on patient management as measured by shortened patient stays, specific therapeutic intervention, reduction of empirical expensive therapy administration, all of which should translate into significant health care savings. OBJECTIVE: The present study aimed at implementing, and evaluating both clinically and analytically the performance of several commercially available PCR based assays for the detection of Herpes virus infections of the CNS. STUDY DESIGN: A total of 314 patients with suspected CNS Herpesvirus infection were investigated, between 1999 and 2001, by Stair primers PCR. Starting on January 2002, two commercially available real-time-PCR systems were implemented and tested using the Stair primers PCR assay as golden standard and three external control proficiency panels along with serial dilutions of positive clinical samples. RESULTS: Sensitivity of the assay was determined to be <200 copies per ml for HSV and <1250 copies per ml for CMV. Positive results were obtained for 17 patients (6 HSV-1, 1 HSV-2, 1 EBV, 1 CMV, 6 VZV and 2 HHV-6) whose clinical and analytical findings were consistent with the PCR results. A real-time-PCR procedure was introduced in 2002 with similar sensitivity, but a more rapid response. CONCLUSION: Conventional end-point PCR proved useful to the diagnosis of CNS herpes virus infection, with an impact on the clinical intervention. However, the use of Real-Time-PCR has greatly enhanced these advantages, making results available at a much earlier time, thus significantly reducing the need for empirical treatment. PMID- 12091083 TI - Detection of herpesvirus genomes by polymerase chain reaction in cerebrospinal fluid and clinical findings. AB - BACKGROUND: The viruses of the Herpesviridae family, in particular herpes simplex virus types 1 (HSV-1) and 2 (HSV-2), cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella zoster virus (VZV), and human herpesvirus 6 (HHV-6), are responsible for numerous infections of the central nervous system (CNS). These infections manifest as diverse clinical signs, many of which are not specific. The diagnosis of these infections is necessary to make it possible to adapt treatment appropriately, as treatment is specific for the particular virus concerned. OBJECTIVES: To apply a polymerase chain reaction (PCR) technique for the diagnosis in a single reaction of the six herpesviruses most frequently detected in the cerebrospinal fluid (CSF) and to analyse clinical events in patients presenting positive results in PCR for herpesviruses. STUDY DESIGN: We studied 141 patients, from whom 180 CSF samples were collected. The clinical files of the patients were consulted retrospectively, and a list of clinical signs was recorded. After testing by targeted PCR, at the clinician's demand, we tested these samples by herpes consensus PCR, which detects six herpesviruses (HSV-1, HSV-2, CMV, EBV, VZV, HHV-6), in a single PCR. RESULTS: Targeted PCR tests identified 25 CSF samples (13.9%), corresponding to 18 patients (12%), as positive. The herpes consensus PCR test detected 49 samples (27.2%) as positive, resulting in the identification of 54 individual viruses (four samples displayed co-infection) from 39 patients (27%). 130 CSF samples, from 101 patients, tested negative by both techniques. 23 HIV-positive patients (30.6%), three HIV-negative immunocompromised patients (27%), and 14 immunocompetent patients (25%) were CSF PCR-positive. In HIV-positive patients, CMV was the virus most frequently identified (13%), followed by EBV (10.6%), VZV (5.3%) and finally HSV-1 and HSV-2 (both 1.3%). We did not detect HHV-6 in any of these samples. We detected only HSV-2, EBV and VZV in the 11 HIV-negative immunocompromised patients. CSF samples of immunocompetent patients contained mostly VZV (9%) and HSV-1 (7.3%). CONCLUSIONS: The herpes consensus PCR for a given virus was more sensitive than the standard, targeted PCR used in our laboratory. The clinical signs presented by patients infected with HSV-1, HSV-2 and CMV were similar to those reported in previous studies. For VZV, we report the possibility of mild, transient cerebral viral reactivation. Our data on the detection of EBV by PCR suggest that the PCR test is of predictive value for cerebral lymphoma in immunocompromised patients. The possible role of HHV-6 in a subacute neurological disorder merits further investigation. PMID- 12091085 TI - Diagnosis of herpesvirus infections of the central nervous system. AB - BACKGROUND: Human herpesviruses may cause infections of the central nervous system (CNS). The early diagnosis of herpesvirus-associated neurological diseases is of high importance. OBJECTIVES: The objective of this paper is to summarize the experience gained with the diagnosis of herpesvirus infections of the CNS at our institute by polymerase chain reaction (PCR)-based assays within the past few years. STUDY DESIGN: A retrospective analysis of herpesvirus desoxy ribonucleic acid (DNA)-positive cerebrospinal fluid (CSF) samples was performed, with particular emphasis on data obtained by quantification of virus DNA in CSF by newly established real-time quantitative PCR assays. RESULTS: Herpesviruses were found in 26.6% of all virus-positive CSF samples detected at our institute between 1995 and 2001. The overall broad testing for different herpesviruses from CSF has led to an increase in the detection rate, especially in relation to varicella zoster virus (VZV)-associated CNS disease. The herpesvirus DNA load in CSF was investigated by TaqMan real-time PCR assays that were established for the individual herpesviruses. The amount of virus varied among the individual diseases, associated with herpes simplex virus type 1, herpes simplex virus type 2, VZV and cytomegalovirus, while for Epstein-Barr virus and human herpesvirus type 6 only low levels of virus were detectable in CSF. CONCLUSIONS: A generally broad testing for different herpesviruses in CSF samples is highly recommended. In addition, determination of the virus DNA level in CSF by quantitative assays seems to be of high importance for elucidating aspects concerning the prognosis of disease, the prediction of distinct CNS manifestations, and possibly the differentiation between specific virus-associated disease and unspecific presence of virus in CSF, especially in immunocompromised patients. PMID- 12091084 TI - Herpes Consensus PCR test: a useful diagnostic approach to the screening of viral diseases of the central nervous system. AB - BACKGROUND: Infections of the central nervous system (CNS) are a difficult diagnostic problem for both clinicians and microbiologists. Various clinical signs, such as encephalitis, myelitis, meningitis, may be associated with herpesviruses. The use of multiplex 'Herpes Consensus' polymerase chain reaction (HC-PCR) in association with nested PCR (nPCR), in addition to classical techniques, made it possible to optimise the management of cerebrospinal fluid (CSF) and serum samples from patients affected by these viral diseases of the CNS. OBJECTIVES: To test by HC-PCR by nPCR and cell culture the CSF and sera from patients with viral infections of the CNS. STUDY DESIGN: We analysed 320 CFS, 154 serum samples and 11 various samples from 286 patients with clinically suspected encephalitis, meningitis or other diseases of the CNS by HC-PCR, nPCR and traditional investigations (cell culture and serological tests). RESULTS: On molecular analysis with the HC-PCR test, 51 CFS samples (15.9%) were positive for at least one of the six target Herpes viruses: fourteen for Herpes simplex 1 (HSV 1), seven for HSV-2, 12 for Cytomegalovirus (CMV; one of which was from an HIV positive patient), five for Epstein-Barr virus (EBV; four of which were from HIV positive patients), three for Varicella-Zoster virus (VZV), five for Human Herpes virus type 6 (HHV-6), three for HSV-1 with HHV-6 co-infection (two cases) and HSV 2 co-infection (one case), and two for HHV-6 with CMV or EBV co-infection (both from patients with immune deficiency). A further 12 samples were positive in nPCR for HHV-7 (8), ADV (1), Enterovirus (1), HSV-1 (1), EBV (1). Of the 154 serum samples, 17 (11.0%) tested positive by HC-PCR for HSV-1 (4), HSV-2 (1), CMV(1), EBV(1), VZV(3) or HHV-6(6), 1 with co-HSV-2/VZV infection. A further five samples tested positive for HHV-7 in nPCR. Culture and tests for antibodies did not supply sufficiently sensitive and specific data. CONCLUSIONS: Our laboratory experience shows that herpesviruses play a central aetiological role in viral infections of the CNS. PCR analysis, especially the HC-PCR test, have revolutionised the diagnostic approach to such infections, making possible rapid, specific and highly sensitive baseline screening. In this way, microbiological investigations can lead to prompt diagnosis, which was limited in the past to a very small number of cases. PMID- 12091086 TI - Herpesviruses and enteroviruses in infections of the central nervous system: a study using time-resolved fluorometry PCR. AB - BACKGROUND: Polymerase chain reaction (PCR) is becoming the new standard for virological diagnosis of the infections of the central nervous system (CNS). Varicella-zoster virus (VZV) has been considered as the leading cause of viral meningitis or encephalitis in Finland, herpes simplex viruses (HSV) and enteroviruses being the next common causative agents. OBJECTIVES: To elucidate the roles of viruses in infections of the CNS by use of novel, sensitive time resolved fluorescence (TRF) PCR assays. STUDY DESIGN: We have utilized TRF PCR assays for diagnostics of HSV, VZV, human cytomegalovirus (CMV) and enteroviruses in infections of the CNS. When relevant, we have also applied virus culture and CSF IgM antibody determinations to elucidate the involvement of other viruses in the CNS infections. The material consisted of CSF samples from hospitals in Western Finland, submitted for diagnostic testing for CNS viral infections during the years 2000-2001. A total of 922 CSF samples were tested by PCR. RESULTS: The PCR assays yielded the virological diagnosis in 72 cases whereas only 24 samples were positive by virus culture or CSF IgM antibody assays. Enteroviruses were found in 6.8%, VZV in 5.8% and HSV in 4.6% of the studied CSF samples. The virus culture and CSF antibody tests yielded 0.5-3.2% positive findings. CONCLUSIONS: The PCR assays of CSF specimens are most effective for the virological diagnosis of CNS infections. In our study, HSV was the most common causative agent, whereas the diagnostic power of TRF-PCR test was highest for enteroviruses. PMID- 12091087 TI - The guide to community preventive services: oral health. PMID- 12091088 TI - Community preventive services and oral health: wishes for the future. PMID- 12091089 TI - Partnering to unlock the mysteries of oral diseases and injuries. PMID- 12091090 TI - Community-based oral health prevention. Issues and opportunities. PMID- 12091091 TI - The Robert Wood Johnson Foundation's response to improving the nation's oral health. PMID- 12091092 TI - Recommendations on selected interventions to prevent dental caries, oral and pharyngeal cancers, and sports-related craniofacial injuries. PMID- 12091093 TI - Reviews of evidence on interventions to prevent dental caries, oral and pharyngeal cancers, and sports-related craniofacial injuries. AB - This report presents the results of systematic reviews of effectiveness, applicability, other positive and negative effects, economic evaluations, and barriers to use of selected population-based interventions intended to prevent or control dental caries, oral and pharyngeal cancers, and sports-related craniofacial injuries. The related systematic reviews are linked by a common conceptual approach. These reviews form the basis of recommendations by the Task Force on Community Preventive Services (the Task Force) about the use of these selected interventions. The Task Force recommendations are presented in this supplement. PMID- 12091094 TI - A comparison of selected evidence reviews and recommendations on interventions to prevent dental caries, oral and pharyngeal cancers, and sports-related craniofacial injuries. PMID- 12091095 TI - Reviews and recommendations to prevent dental caries, oral and pharyngeal cancers, and sports-related craniofacial injuries. PMID- 12091096 TI - The acyl composition of mammalian phospholipids: an allometric analysis. AB - Data concerning the acyl composition of tissue phospholipids from mammal species, ranging in size from the shrew (7 g) to cattle (370 kg), has been collated from the literature and analysed allometrically. Phospholipids from heart, skeletal muscle, liver and kidney exhibited similar allometric trends whereby phospholipids had a significant decrease in unsaturation index (number of double bonds per 100 acyl chains) as species body size increased whilst there was no change in the percent of unsaturated acyl chains. Whilst total polyunsaturate content did not change with body mass, both heart and skeletal muscle phospholipids showed a significant allometric decrease in the omega-3 polyunsaturate content. The content of the highly polyunsaturated docosahexaenoic acid (22:6 n-3) in phospholipids showed significant and substantial allometric decline with increasing body mass in all four tissues (exponents ranged from 0.19 in liver to -0.40 in skeletal muscle). Brain phospholipids showed no allometric trends in acyl composition and were highly polyunsaturated in all species. These trends are discussed in light of the hypothesis that the relative content of polyunsaturated acyl chains in membranes, and especially docosahexaenoate (22:6 n-3), can act as a membrane pacemaker for metabolic activity. PMID- 12091097 TI - Actions of two serine proteases from Trimeresurus jerdonii venom on chromogenic substrates and fibrinogen. AB - Jerdonobin and jerdofibrase are two serine proteases purified from the venom of Trimeresurus jerdonii. The Michaelis constant K(m) and the catalytic rate constant K(cat) of jerdonobin or jerdofibrase on three chromogenic substrates, H D-Pro-Phe-Arg-pNA (S2302), H-D-Phe-pipecolyl-Arg-pNA (S2238), and H-D-Val-Leu-Lys pNA (S2251) were obtained from lineweaver-Burk plots. Jerdofibrase could hydrolyze all three substrates, but jerdonobin had no detectable activity on S2251, suggesting a relatively broader substrate specificity for jerdofibrase than jerdonobin. By SDS-PAGE, jerdofibrase preferentially degraded Bbeta-chain of fibrinogen. It also degraded Aalpha-chain of fibrinogen with relatively slow activity, but did not act on the gamma-chain. In contrast, jerdonobin did not degrade fibrinogen within 12 h. Fibrinopeptides liberation test, identified by HPLC, showed jerdonobin released fibrinopeptide A and a small amount of fibrinopeptide B. Unlike jerdonobin, jerdofibrase mainly released fibrinopeptide B. These results indicate that the two enzymes differ in their ability to hydrolyze chromogenic substrates and in their actions on fibrinogen. PMID- 12091098 TI - Occurrence of a novel NADP(+)-linked alcohol dehydrogenase in Euglena gracilis. AB - An NADP(+)-dependent alcohol dehydrogenase was found in Euglena gracilis Z grown on 1-hexanol, while it was detected at low activity in cells grown on ethanol or glucose as a carbon source, indicating that the enzyme is induced by the addition of 1-hexanol into the medium as a carbon source. This enzyme was extremely unstable, even at 4 degrees C, unless 20% ethylene glycol was added. The optimal pH was 8.8-9.0 for oxidation reaction. The apparent K(m) values for 1-hexanol and NADP(+) were found to be 6.79 mM and 46.7 microM for this enzyme, respectively. The substrate specificity of this enzyme was very different from that of already purified NAD(+)-specific ethanol dehydrogenase by showing the highest activity with 1-hexanol as a substrate, followed by 1-pentanol and 1-butanol, and there was very little activity with ethanol and 1-propanol. This enzyme was active towards the primary alcohols but not secondary alcohols. Accordingly, since the NADP(+)-specific enzyme was separated on DEAE cellulose column, Euglena was confirmed to contain a novel enzyme to be active towards middle and long-chain length of fatty alcohols. PMID- 12091099 TI - Galactose-containing polysaccharides from Dictyostelium mucoroides as possible acceptor molecules for cell-type specific galactosyl transferase. AB - Dictyostelium discoideum has polysaccharides that accept galactose residues by the action of cell-type-specific galactosyl transferase. This paper describes partial purification of the major galactose-accepting polysaccharide that was isolated from the related strain, Dictyostelium mucoroides and proposes its plausible carbohydrate sequences. The most potent acceptor activity was observed in the neutral and Ricinus communis agglutinin (RCA-60) bound galactosaminoglycan, consisting of galactose (Gal) and galactosamine (GalN). However, the acceptor polysaccharides are highly heterogeneous in the reactivity with RCA-120 and in molecular mass. The peak fraction was analyzed by (1)H- and (13)C-NMR studies, methylation analysis, beta-D-galactosidase digestion, controlled Smith degradation and reducing terminal analysis. Based on these results, we tentatively propose the following novel type of the common carbohydrate sequences as the acceptor polysaccharides. [abstract - see text]. PMID- 12091100 TI - Purification and characterization of a 28-kDa major protein from ginseng root. AB - A major protein was isolated from ginseng root (Panax ginseng C.A. Meyer) using a combination of ammonium sulfate fractionation, gel filtration chromatography, ion exchange FPLC, and fast performance liquid chromatofocusing. Electrophoretic and gel permeation chromatographic studies revealed that the major protein, GMP, is composed of two subunits of approximately 28 kDa. During purification, it was found that the elution profiles of GMP from gel filtration chromatography were significantly different, depending on the ionic strength of buffers used. GMP in a buffer of low ionic strength was isolated as a complex with carbohydrate, which could be only dissociated at high ionic strength. Carbohydrate composition in GMP detected by gas chromatography varied, depending on the isolation method of the protein from ginseng roots. These results suggest that carbohydrates are bound non-covalently to GMP whose amino acid composition analysis showed high amounts of acidic amino acids. PMID- 12091101 TI - The esterification and modification of n-3 and n-6 polyunsaturated fatty acids by hepatocytes and liver microsomes of turbot (Scophthalmus maximus). AB - The present study was undertaken to establish whether the formation of 22:6n-3 from 18:3n-3 and/or 20:5n-3 can occur in turbot liver and if this conversion is consistent with the operation of a Delta4 desaturase-independent pathway. At the same, time the effects of feeding a diet devoid of long chain polyunsaturated fatty acids (PUFA) on the patterns of esterification and modification of 18:3n-3, 20:5n-3 and 18:2n-6 by turbot hepatocytes and liver microsomes were examined. For this purpose, two groups of fish (25-30 g) were employed: one was fed a commercial diet containing fish oil (FO) and thus rich in long chain n-3 PUFA and the other was fed an experimental diet based on olive oil (OO). After 5 months of feeding, hepatocytes and liver microsomes isolated from individuals in the two groups of fish were incubated with [1-(14)C]-PUFA [either 18:3n-3, 20:5n-3 or 18:2n-6]. After 3 h of incubation, most radioactivity from all three radiolabelled substrates incorporated into lipids by hepatocytes and microsomes was recovered in the original substrate. The formation of desaturation products of n-3 radiolabelled substrates was higher in hepatocytes isolated from OO-fed than FO-fed fish. Small amounts of radiolabelled 22:6n-3 were formed from [1 (14)C]18:3n-3 and [1-(14)C]20:5n-3, but only by hepatocytes from fish fed OO, which also synthesised a small amount of radiolabelled 24:6n-3 from 14C-20:5n-3. Elongation products predominated over desaturation products in hepatic microsomes from both groups of fish studied, particularly in microsomes from fish fed FO. The results confirm that regardless of the long chain PUFA content of the diet, the production of 22:6n-3 in turbot liver from 18:3n-3 and/or 20:5n-3, and of 20:4n-6 from 18:2n-6, is very limited. The presence of radiolabelled 24:6n-3 in microsomes coupled with the absence of radiolabelled 22:6n-3 suggests that the formation of 22:6n-3 that does occur in turbot liver cells, may involve C24 intermediates and peroxisomal beta-oxidation. PMID- 12091102 TI - Cloning of a novel phospholipase A2 from the cnidarian Adamsia carciniopados. AB - PLA2 catalytic activity was detected in homogenised tissues, including tentacles and acontia (structures for preying and defence, respectively), of the sea anemone Adamsia carciniopados. Nested reverse transcription polymerase chain reaction (RT PCR) with degenerate primers and rapid amplification of cDNA ends (RACE) were used to clone a novel phospholipase A2 from Adamsia carciniopados (AcPLA2). AcPLA2 contains a putative prepropeptide of 37 residues, ending with a basic doublet followed by a mature protein of 119 amino acids, including 12 cysteines. AcPLA2 displays only 30-42% similarity with other known secretory PLA2s (sPLA2). C-terminal extension, typical of groups II and X PLA2s, is absent. Predicted molecular weight and pI of the mature protein are 13.5 kDa and 9.1, respectively. Structural features and phylogenetic analysis set AcPLA2 apart from the known sPLA2s and define this molecule in the ancient metazoan phylum Cnidaria as a member of a new class of sPLA2s. PMID- 12091103 TI - Immunological study of muscle parvalbumin isotypes in three African catfish during development. AB - Eleven parvalbumin isotypes expressed during the development of clariids Heterobranchus longifilis and Clarias gariepinus and claroteid Chrysichthys auratus were purified and electrophoresed on sodium-dodecyl-sulfate polyacrylamide gels. Immunochemical cross-reactions among these proteins were investigated by immunoblotting, using purified antibodies raised against three isotypes chosen at different stages of fish development. Antibodies raised against H. longifilis PA I (larval-juvenile isotype) and against C. gariepinus PA IIIa (juvenile-adult isotype) cross-reacted to a rather similar extent despite a weaker cross-reaction of anti-PA IIIa with larval-juvenile isotypes. On the other hand, antibodies raised against H. longifilis PA IV (an exclusively adult isotype) recognized markedly only H. longifilis PA IV and C. gariepinus PA IIIb. These two adult isotypes most likely belong to the alpha lineage, and all the others to the beta lineage. These results show that parvalbumin isotypes synthesized at different stages of fish growth differ structurally, and that the most marked difference is between larval-juvenile and adult clariid isotypes. PMID- 12091104 TI - Molecular characterization of the bifunctional VHDL-CP from the hemolymph of white shrimp Penaeus vannamei. AB - A very high-density lipoprotein (VHDL) purified from the hemolymph of the white shrimp Penaeus vannamei is shown to be identical to the clotting protein (CP) previously reported from the same organism based on size, subunits and N-terminal amino acid sequence. The approximately 440-kDa protein, a homodimer of approximately 200-kDa subunits, was present in KBr gradient fractions ranging in density from 1.155 to 1.212 g/ml. Samples of VHDL after purification by strong cation exchange chromatography were subjected to electrophoresis on native polyacrylamide gels. Lipids associated with the VHDL were detected by Sudan Black and Oil Red O staining and comprise 9-15% of the purified protein. Circular dichroism of VHDL-CP indicates that the alpha-helix content of the VHDL-CP is 32%, while beta-sheets correspond to 33%, closely resembling the secondary structure of CP from the shrimp Penaeus monodon and, remarkably, the secondary structure of very high-density lipophorin E (VHDLpE) from the tobacco hornworm, Manduca sexta. PMID- 12091105 TI - Digestive proteinases of Artemesia longinaris (Decapoda, Penaeidae) and relationship with molting. AB - Digestive proteinase activities of Artemesia longinaris were assayed at different stages of the molting cycle. Total proteolytic activity in the hepatopancreas was highest during postmolt. Trypsin and chymotrypsin activities were highest during intermolt. Specific inhibitors and zymograms of A. longinaris hepatopancreas extracts showed four trypsins (14.79, 15.49, 16.60, 17.38 kDa, respectively) and three chymotrypsins (21.38, 22.91, 27.54 kDa, respectively). Our results suggest that proteolytic activity in the hepatopancreas of A. longinaris is influenced by the molting cycle. Types and activity of prawn digestive enzymes constitute background information to further study the digestive abilities of these organisms and will lead to understanding their nutritional needs and feeding ecology. PMID- 12091106 TI - Purification of serum precursor proteins to vitelline envelope (choriogenins) in masu salmon, Oncorhynchus masou. AB - Three vitelline envelope-related proteins (VERPs), very-high-molecular-weight VERP (vhVERP), high-molecular-weight VERP (hVERP) and low-molecular-weight VERP (lVERP) were purified from female masu salmon serum. The apparent molecular weights of vhVERP, hVERP and lVERP, in their native state, were 520, 88 and 54 kDa, respectively, by gel-filtration chromatography. Very-high-molecular-weight VERP comprises two subunits, corresponding to 175 and 126 kDa. On SDS-PAGE, hVERP and lVERP migrate at 53 and 47 kDa, respectively. Amino acid analysis of vhVERP and hVERP showed that they share a high content of glutamic acid and proline. By contrast, lVERP is rich in glutamic acid and asparatic acid. These features are in good agreement with the amino acid composition of the vitelline envelope. Immuno-biochemical analysis suggested that vhVERP is derived from hVERP by polymerization and/or aggregation. Antibodies against hVERP and lVERP specifically immunostained the vitelline envelope and liver of female masu salmon. In addition, both hVERP and lVERP were induced in the serum of estrogen treated male fish. Taken together, it is suggested that hVERP and lVERP are homologous molecules with choriogenin H and choriogenin L in medaka, respectively. These results indicate that hVERP and lVERP are precursor proteins to the vitelline envelope (choriogenins) in masu salmon. PMID- 12091107 TI - Chemical characterization of the oligosaccharides in beluga (Delphinapterus leucas) and Minke whale (Balaenoptera acutorostrata) milk. AB - Carbohydrates were extracted from the milk of a beluga, Delphinopterus leucas (family Odontoceti), and two Minke whales, Balaenoptera acutorostrata (Family Mysticeti), sampled late in their respective lactation periods. Free oligosaccharides were separated by gel filtration and then neutral oligosaccharides were purified by preparative thin layer chromatography and gel filtration, while acidic oligosaccharides were purified by ion-exchange chromatography, gel filtration and high performance liquid chromatography (HPLC). Their structures were determined by 1H-NMR. In one of the Minke whale milk samples, lactose was a dominant saccharide, with Fuc(alpha1-2)Gal(beta1-4)Glc(2' fucosyllactose), Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Glc(lacto-N-neotetraose), GalNAc(alpha1-3)[Fuc(alpha1-2)]Gal(beta1-4)Glc(A-tetrasaccharide), Gal(beta1 4)GlcNAc(beta1-3)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Glc (para lacto-N neohexaose), Neu5Ac(alpha2-3)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Glc (sialyl lacto-N-neotetraose), Neu5Ac(alpha2-6)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)Glc (LST c) and Neu5Ac(alpha2-3)Gal(beta1-4)GlcNAc(beta1-3)Gal(beta1-4)GlcNAc(beta1 3)Gal(beta1-4)Glc (sialyl para lacto-N-neohexaose) also being found in the milk. The second Minke whale sample contained similar amounts of lactose, 2' fucosyllactose and A-tetrasaccharide, but no free sialyl oligosaccharides. Sialyl lacto-N-neotetraose and sialyl para lacto-N-neohexaose are novel oligosaccharides which have not been previously reported from any mammalian milk or colostrum. These and other oligosaccharides of Minke whale milk may have biological significance as anti-infection factors, protecting the suckling young against bacteria and viruses. The lactose of Minke whale milk could be a source of energy for them. The beluga whale milk contained trace amounts of Neu5Ac(alpha2 3)Gal(beta1-4)Glc(3'-N-acetylneuraminyllactose), but the question of whether it contained free lactose could not be clarified. Therefore, lactose may not be a source of energy for suckling beluga whales. PMID- 12091109 TI - Putative osmolytes in the kidney of the Australian brush-tailed possum, Trichosurus vulpecula. AB - The Australian brush-tailed possum, Trichosurus vulpecula, is capable of producing a moderately concentrated urine, at least up to 1300 mOsm l(-1). Kidneys of adult animals fed in captivity on a normal diet with ready access to water were analysed. The inner medullary regions were found to have moderately high concentrations of sodium (outer medulla, 367+/-37; inner medulla 975+/-93 mmol kg(-1) dry wt.), chloride (outer medulla 240+/-21; inner medulla 701+/-23 mmol kg(-1) dry wt.) and urea (outer medulla, 252+/-62; inner medulla, 714+/-69 mmol kg(-1) protein). When the animals were fed on a 'wet diet', amounts of these substances in the outer medulla and cortex were reduced, although with the exception of urea these changes were not significant. There were highly significant changes in amounts of Na(+), Cl(-) and urea in the inner medulla (Na(+), 566+/-7; Cl(-), 422+/-9 mmol kg(-1) dry wt.; urea 393+/-84 mmol kg(-1) protein). Likewise, the inner medulla of animals fed a 'dry diet' with limited access to water showed highly significant increases in the same substances (Na(+), 1213+/-167; Cl(-), 974+/-137 mmol kg(-1) dry wt.; urea, 1672+/-98 mmol kg(-1) protein). Inositol was found in the outer medulla (224+/-90 mmol kg(-1) protein) and inner medulla (282 mmol kg(-1) protein) as was sorbitol (outer medulla, 62+/-20; inner medulla, 274+/-72 mmol kg(-1) protein). Both these polyols were reduced in amount in renal tissue from 'wet diet' animals, and increased in 'dry diet' animals, but the changes were not statistically significant. The methylamines, betaine and glycerophosphorylcholine (GPC), showed a similar pattern, but both were significantly elevated in the inner medulla of 'dry diet' animals (betaine 154+/-57 to 315+/-29 mmol kg(-1) protein; GPC 35+/-7 to 47+/-10 mmol kg(-1) protein). It was concluded that in this marsupial the concentrating mechanism probably functions in a similar way to that in higher mammals, and that the mechanism of osmoprotection of the medulla of the kidney involves the same osmolytes. However, the high ratio of betaine to GPC in the inner medulla resembles the situation in the avian kidney. PMID- 12091108 TI - Comparison of lipids in selected tissues of the Florida manatee (Order Sirenia) and bottlenose dolphin (Order Cetacea; Suborder Odontoceti). AB - The position, porosity and oil-filled nature of the zygomatic process of the squamosal bone (ZPSB) of the Florida manatee, Trichechus manatus latirostris, suggest that it may have a similar sound conduction function to that of the intramandibular fat body (IMFB) of the bottlenose dolphin, Tursiops truncatus, and other odontocetes. To examine this possibility we determined the lipid composition of the ZPSB and adipose tissue from the dorsal part of the head region of the Florida manatee, and compared it to that of the dolphin IMFB and melon (another fatty area implicated in sound conduction in odontocetes). Lipids from manatee ZPSB and from adipose tissue were composed almost entirely of triacylglycerols. The most abundant fatty acids of the ZPSB were 18:1, 16:0, 14:0 and 16:1. The major fatty acids of the adipose tissue in the head were the four mentioned above, along with 12:0 and 18:0. Manatee samples did not contain isovaleric acid (iso-5:0), which was found in the bottlenose dolphin IMFB and melon, and has been related to sound conduction in dolphins and some other odontocetes. Thus, if manatee tissues are capable of sound conduction, and this process does occur through the ZPSB, a somewhat different suite of lipid components must support this function. PMID- 12091110 TI - The effects of hypoosmotic infusion on the composition of renal tissue of the Australian brush-tailed possum Trichosurus vulpecula. AB - Although the occurrence of organic osmolytes in the inner medulla of the marsupial kidney has been recently reported [Comp. Biochem. Physiol. (2002) 132B 635-644], changes in these substances, in response to water loading in vivo, has not been studied. Adult Trichosurus vulpecula, the Australian brush-tailed possum, were subjected to water deprivation for 48 h. Following anaesthesia and unilateral nephrectomy, the animals were perfused with hypo-osmotic saline (80 mmol l(-1); 1.5 ml min(-1)) for 60 min. This resulted in a rapid increase in urine volume and a corresponding fall in urine osmolality. At the end of the infusion the animals were killed and the second kidney removed. Analysis of the renal tissue revealed that water content of cortical, outer and inner medullary regions of the kidney increased slightly following infusion, while sodium, and chloride contents of all three regions fell. Potassium contents, on the other hand, were barely changed. Of the organic osmolytes determined, very significant decreases in the inner medulla, following infusion, were found for sorbitol (from 397+/-79 to 266+/-49 mmol kg(-1) protein), inositol (247+/-23 to 190+/-25 mmol kg(-1) protein), and betaine (464+/-70 to 356+/-21 mmol kg(-1) protein), while only inositol was significantly decreased in the outer medulla (197+/-22 to 150+/ 16 mmol kg(-1) protein). Glycerophosphorylcholine levels were low throughout the kidney and were not significantly affected by the infusion. It was concluded that inositol and sorbitol play a significant role as compatible organic osmolytes in the possum kidney, while betaine functions as the principal counteracting osmolyte. Amino acid levels in the cortex and outer medulla showed no overall change in amount following infusion, although there were highly significant changes in individual amino acids. In the inner medulla there was a highly significant reduction in total amino acids with infusion, largely due to a fall in amounts of taurine (104+/-4 to 75+/-17 mmol kg(-1) protein), and glycine (97+/ 15 to 71+/-18 mmol kg(-1) protein). A fall in free amino acid levels in the inner medulla appears to significantly contribute to the process of intracellular osmotic adjustment during an induced diuresis. PMID- 12091111 TI - Color changes in Halyomorpha brevis (Heteroptera: Pentatomidae) correlated with distribution of pteridines: regulation by environmental and physiological factors. AB - Halyomorpha brevis (Heteroptera: Pentatomidae) produces adults with different color patterns, most noticeable in the sternum. The color, ranging from ivory to red, depends on the extent of the accumulation of red pigment. The present work investigated the effects of photoperiod, temperature and aging on the pigmentation. The red pigment was identified as erythropterin by comparing the Rf with standard pteridines in paper chromatography in three solvent systems. Erythropterin was found in all organs, red or light red. Uric acid was detected prominently in ivory or light red sternum. A negative correlation was found between the extent of red pigmentation and that of uric acid content. The relative proportion of males and females with a red sternum progressively increased as they aged from day 0 to day 20 after adult emergence, particularly in males reared under a long day (non-diapausing). In males, an age-dependent decrease in red pigmentation was observed in the gastric ceca and tracheae. The sternum was lighter in short-day adults (diapausing) than that in non-diapausing adults at the same age, and the latter never achieved the intense red color. The possible functions of pteridines are discussed. PMID- 12091112 TI - Molecular cloning and expression of a cDNA encoding the luciferase from the firefly, Hotaria unmunsana. AB - We have cloned and characterized a cDNA encoding the luciferase from the firefly, Hotaria unmunsana. The cDNA encoding the luciferase of H. unmunsana was isolated by RT-PCR with a gene-specific primer set. The cDNA encoding the luciferase of H. unmunsana is 1644 base pairs long with an open reading frame of 548 amino acid residues. The deduced amino acid sequence of the luciferase gene of H. unmunsana showed 98.0 and 96.8% identity to H. parvula and Luciola mingrelica, respectively. Phylogenetic analysis further confirmed that the deduced amino acid sequences of the H. unmunsana luciferase gene belonged to the same subfamily, Luciolinae. Southern blot analysis of genomic DNA suggested the presence of the H. unmunsana luciferase gene as a single copy and Northern blot analysis confirmed light organ-specific expression at the transcriptional level. The cDNA encoding the luciferase of H. unmunsana was expressed as a 61-kDa band in the baculovirus-infected insect cells and the extracts of the recombinant baculovirus infected cells emitted luminescence in the luciferase activity assay. PMID- 12091113 TI - Localization of group IB phospholipase A(2) isoform in the gills of the red sea bream, Pagrus (Chrysophrys) major. AB - We previously reported that PLA(2) activity in the gills is higher than that in other tissues in red sea bream and purified PLA(2) from the gills belongs to the group IB PLA(2) as well as other red sea bream PLA(2)s. In this study, we reconfirmed that the level of PLA(2) activity is extremely high in the gills compared with other tissues, and gill PLA(2) was detected only in the gills by immunoblotting and inhibition test using anti-gill PLA(2) monoclonal antibody. The level of PLA(2) activity and protein expression in the gills are well correlated. Fish can be roughly divided into high and low groups based on the level of PLA(2) activity. Gill PLA(2) was detected in the gills of the high group, but not the low group by immunoblotting. In the gills of the high group, gill PLA(2) was detected in the mucous cells and pavement cells located on the surface of gill epithelia by immunohistochemistry. On the other hand, positive signals were observed only in the mucous cells by in situ hybridization. We also isolated inactive proPLA(2), having AR propeptide, preceding the mature enzyme from the gill extract. These results suggest that gill PLA(2) is synthesized as an inactive proPLA(2) in the mucous cells and is secreted to the surface of gill epithelia. PMID- 12091114 TI - Thromboprophylaxis in acute medical patients: need for an implementation strategy. PMID- 12091115 TI - Is there any role left for p210-derived peptide vaccines in chronic myeloid leukemia? PMID- 12091116 TI - A milestone in the study of the vascular system: Wilhelm Roux's doctoral thesis on the bifurcation of blood vessels. PMID- 12091117 TI - FK506 in the maturation of dendritic cells. AB - BACKGROUND AND OBJECTIVES: FK506 (tacrolimus) is a potent immunosuppressive agent that inhibits interleukin-2 (IL-2) and interferon-g production by CD4+ cells. The effect of this agent on dendritic cells (DCs), the highly professional antigen presenting cells for T-cells, has not been completely defined. We investigated the effect of FK506 on DC differentiation from monocytes, and on the shift from immature to mature immunophenotypes. DESIGN AND METHODS: DCs were generated in vitro from monocytes of healthy donors. Cells were exposed to lipopolysaccharide (LPS) and two doses of FK506, with variations in time of exposure and sequence of FK506 and LPS addition. Immunophenotype analysis in immature and mature DCs under FK506 treatment was performed by flow cytometry at the end of cell culture. The Student's t-test was used for statistical analyses. RESULTS: FK506 did not affect dendritic cell generation or viability. There were no changes in cell surface markers with addition of FK506 at physiologic concentrations (10 ng/mL). We found a decrease in CD1a median fluorescence intensity (MFI) and an increase in percentage of CD86-positive cells with lengthy exposure (6 days) to FK506 at 5000 ng/mL. In the sequential study, 5000 ng/mL FK506 before LPS addition resulted in a significant decrease in CD1a MFI and in the percentage of cells co-expressing CD83 and CD86. INTERPRETATION AND CONCLUSIONS: Our results indicate that lengthy exposure to 5000 ng/mL FK506 modified the expression of some DC-cell surface markers, maintaining DCs in a low maturity stage. PMID- 12091118 TI - Novel types of bcr-abl transcript with breakpoints in BCR exon 8 found in Philadelphia positive patients with typical chronic myeloid leukemia retain the sequence encoding for the DBL- and CDC24 homology domains but not the pleckstrin homology one. AB - BACKGROUND AND OBJECTIVES: We previously described a novel type of the chimeric bcr-abl mRNA transcript in a patient with a Philadelphia chromosome positive chronic myeloid leukemia. A similar bcr-abl transcript has also been described by others. DESIGN AND METHODS: Sequence analysis of the fusion region showed a join between part of exon e8 of the bcr gene and an intronic sequence of abl intron 1b spliced on exon a2 of the abl gene, giving rise to an in-frame e8-int-a2 bcr-abl transcript, translated into a 197.5 kDa BCR-ABL protein of 1804 amino acid residues, which we named p200 BCR-ABL. RESULTS: In this work, employing protein comparison analysis (pFAM) we show that these novel bcr-abl transcripts retain the DBL homology (DH) domain and the recently recognized CDC24 homology domain, but not the pleckstrin homology (PH) domain of the bcr gene. INTERPRETATION AND CONCLUSIONS: This observation, along with the myeloid immunophenotype of the tumor and, at least in one case, the patient's correspondingly good response to alpha-interferon therapy, suggests that p200 BCR-ABL is more similar to p210 BCR ABL, in which the DH, CDC24 and PH domains are all maintained, than to p185, in which these domains are all lost. PMID- 12091119 TI - Safety and efficacy of subcutaneous Campath-1H for treating residual disease in patients with chronic lymphocytic leukemia responding to fludarabine. AB - BACKGROUND AND OBJECTIVES: Recent observations suggested that targeted monoclonal antibodies might be best employed in lymphoid malignancies under conditions of minimal residual disease. This prompted us to investigate the role of Campath-1H as treatment for patients with chronic lymphocytic leukemia (CLL) in whom fludarabine (FAMP) had produced a marked disease debulking with persistence of bone marrow (BM) infiltration or a complete remission (CR) without the disappearance of the molecular aberration (IgH monoclonal expression). As intravenous Campath-1H is almost invariably associated with reactions, sometimes of WHO grade 3-4, we adopted the subcutaneous route of administration, which proved to induce rare and mild adverse reactions but had comparable efficacy. DESIGN AND METHODS. Nine patients (7 males, 2 females) with a median age of 55 years (range 41-61) who responded to FAMP (1 had a CR, 5 a nodular partial remission [PRN], and 3 a partial remission [PR]), according to NCI Working Group Criteria, received subcutaneous Campath-1H, three times a week for 6 weeks in escalating doses up to 10 mg. Monoclonal rearrangement of IgH was present in all patients before immunotherapy. Patients received acyclovir and cotrimoxazole as infection prophylaxis. Granulocyte colony-stimulating factor (G-CSF), at the dosage of 5-10 microg/kg/die, or intermediate-dose Ara-C (800 mg/m(2)/q 12h x 6 doses), was administered to obtain peripheral blood stem cell (PBSC) mobilization. RESULTS: All patients were evaluable for response. Five patients, 2 in PR and 3 in PRN after FAMP treatment, reached a CR. Three patients, one in PR, one in PRN and one in CR, converted to a molecular remission. In four out of seven patients PBSC harvesting was successful; more than 2.5 x 10(6) cells/kg were collected from all these patients. Collection was polyclonal for IgH gene rearrangement in three cases. One patient has been transplanted after cyclophosphamide and total body irradiation as conditioning regimen, without complications and with rapid hemopoietic engraftment. All patients were evaluable for toxicity. A WHO grade 1-2 skin reaction was observed in 5 patients at the site of injection. No infectious episodes were recorded. Two out of three patients presenting cytomegalovirus reactivation, without pneumonia, were successfully treated with oral gancyclovir. INTERPRETATION AND CONCLUSIONS: Subcutaneous Campath-1H administered to CLL patients with residual BM disease after FAMP proved to be safe and effective. Of nine patients, three obtained a molecular CR and five converted into a morphologic and immunophenotypic CR. In four of seven patients submitted to PBSC mobilization, this treatment also allowed a harvest uncontaminated by CD5/CD19 double-positive CLL cells, which was polyclonal for IgH gene rearrangement in three cases. PMID- 12091120 TI - The prognostic significance of beta(2)-microglobulin in patients with Hodgkin's lymphoma. AB - BACKGROUND AND OBJECTIVES: Serum beta(2)-microglobulin (s beta(2)m) is an established prognostic factor for multiple myeloma and non-Hodgkin's lymphoma, but only limited data suggest an adverse prognostic significance for Hodgkin's lymphoma (HL). This study was undertaken to examine the impact of s beta(2)m on the prognosis of patients with HL. DESIGN AND METHODS: s beta(2)m was measured by a radioimmunoassay (upper normal limit 2.4 mg/L), in pretreatment serum samples of 232 patients with HL, who were then treated with ABVD or equivalent regimens with or without radiotherapy. Multivariate survival analysis was based on Cox's proportional hazards model. RESULTS: Main patients' characteristics: median age 30.5 years (14-78); 58% males; 68% nodular sclerosis, 20% mixed cellularity and 12% lymphocyte predominance; 34% B-symptoms; 24% Ann Arbor stage I, 49% II, 18% III and 9% IV. Elevated s beta(2)m levels were detected in 65/232 patients (28%) and correlated with older age (p<0.001), mixed cellularity (p=0.03), B-symptoms (p=0.002), advanced stage (p=0.02), > or = 5 involved sites (p=0.02), inguinal/iliac involvement (p=0.009), lymphocytopenia (p=0.002) and elevated lactate dehydrogenase (p=0.01). The 7-year failure free survival (FFS) was 75% vs. 72% for patients with normal vs. elevated s beta(2)m (p=0.15). The corresponding 7-year overall survival (OS) rates were 86% vs. 52% (p=0.003). In multivariate analysis, elevated s beta(2)m was not predictive of FFS, but was independently associated with inferior OS (p=0.01), along with the number of involved sites (p<0.001). INTERPRETATION AND CONCLUSIONS: s beta(2)m is not a potent prognostic factor for FFS in optimally treated patients with HL. However s beta(2)m may be predictive of OS, probably due to its effect on the timing of treatment failure. PMID- 12091121 TI - Polymorphisms within glutathione S-transferase genes in pediatric non-Hodgkin's lymphoma. AB - BACKGROUND AND OBJECTIVES: Glutathione S-transferases (GSTs) are involved in the metabolism of a number of cancer chemotherapeutic agents. Certain members within the GST superfamily exhibit phenotypically relevant genetic polymorphisms which have been associated with outcome in hematologic malignant disease. DESIGN AND METHODS: In the present study we genotyped a cohort of 169 pediatric non Hodgkin's lymphoma (NHL) patients with available specimens from the NHL-BFM trials 86 and 90 conducted by the Berlin-Frankfurt-Munster (BFM) study group to assess a potential association of phenotypically relevant glutathione S transferase polymorphisms (GSTM1, GSTT1, GSTP1 codon 105) with treatment outcome in this patient group. RESULTS: Treatment failure in patients with mature B-cell NHL was significantly less likely to occur in patients carrying at least one GSTM1 allele in comparison to those with a homozygous deletion of GSTM1. This protective effect mediated by the presence of GSTM1 was even more pronounced within the subset of therapy group B patients at highest clinical risk of treatment failure (B-ALL, disease stage IV, disease stage III with unresected abdominal tumor, and LDH activity > or = 500 U/L). Of all events in therapy group B, 87.5% occurred in this high risk group. Within this subset, the multivariate relative risk reached 4.98 (95% CI = 1.27-19.52; p= 0.021). INTERPRETATION AND CONCLUSIONS: Our results suggest that genetic variation at the GSTM1 locus may be of clinical importance in pediatric NHL and may be a potential candidate for indicating future treatment stratification strategies. PMID- 12091122 TI - High prevalence of hepatitis G virus infection in Hodgkin's disease and B-cell lymphoproliferative disorders: absence of correlation with hepatitis C virus infection. AB - BACKGROUND AND OBJECTIVES: During the last decade an epidemiological association between hepatitis C virus (HCV) and B-cell lymphoproliferative disorders (B-LPD) has been reported; the same association has not been observed for Hodgkin's disease (HD). Hepatitis G virus (HGV) shares genetic and biological features with HCV, thus it might also be involved in lymphomagenesis. DESIGN AND METHODS: The aim of this study was to compare the prevalence of HCV and HGV infection in patients at diagnosis of B-LPD or HD. RESULTS: We tested 227 consecutive untransfused patients (127 with B-LPD and 100 with HD) and 110 healthy controls. The prevalence of HCV infection was significantly higher in B-LPD patients than in controls (17.3% vs. 1.8%, p<0.002 ), whereas it was the same in HD patients as in controls. In contrast, the prevalence of HGV was significantly higher in patients, both those with B-LPD (7.8% vs. 0.9%, p<0.03) and those with HD (13% vs. 0.9%, p<0.002), than in controls. Among the various B-LPD tested, HGV infection was more frequent in B-NHL (11.5%). INTERPRETATION AND CONCLUSIONS: Our data support the hypothesis that HGV infection may play a role in lymphomagenesis and that this role is different and separate from that of HCV. PMID- 12091123 TI - The cost of treating relapsed indolent non-Hodgkin's lymphoma in an international setting: retrospective analysis of resource use. AB - BACKGROUND AND OBJECTIVES: Few economic data exist on the treatment of indolent non-Hodgkin's lymphoma (NHL) and there are none in the published literature concerning relapsed disease. This international analysis (Canada, Germany, Italy) was established to estimate the overall direct cost of treating patients with relapsed indolent NHL and determine the main cost components of treatment. DESIGN AND METHODS: Telephone interviews were used to identify the most commonly used treatment regimens in each country. CHOP, CVP and fludarabine were chosen for economic analysis, which was based on retrospective data from 424 patients. RESULTS: Overall treatment costs for a course of six cycles varied more than 5 fold, from 3,445 to 17,940 Euros between regimens and countries. The treatment setting had a major impact on costs, with in-patient costs being up to three times greater than the equivalent out-patient values. Drug administration costs comprised 46-60% of the overall treatment costs in the in-patient setting. Adverse event management was the major cost component for out-patient CHOP and CVP therapy (52-75%), and a significant proportion (24-40%) of in-patient costs for these regimens. Drug acquisition accounted for less than half of treatment costs for most of the regimens analyzed. INTERPRETATION AND CONCLUSIONS: This study shows that not simply drug acquisition costs, but the costs of drug administration, particularly in the in-patient setting, and adverse event management are major contributors to the overall treatment costs for relapsed indolent NHL. PMID- 12091124 TI - Synergistic activation of human platelets by lysophosphatidic acid and adrenaline. AB - BACKGROUND AND OBJECTIVES: Platelet reactivity is regulated by various important bioactive and physiologic substances. The objective of this study was to characterize lysophosphatidic acid (LPA)-triggered responses in human platelets. In addition, the effect of LPA was compared with that of other activators and possible synergistic interactions were evaluated. DESIGN AND METHODS: LPA triggered cytosolic Ca(2+) responses were measured using fura-2-loaded platelets in a spectrofluorometer. Furthermore, platelet aggregation and secretion were analyzed in a lumi-aggregometer and protein tyrosine phosphorylation was detected with the Western blot technique. RESULTS: LPA dose-dependently increased cytosolic Ca(2+) concentration ([Ca(2+)](i)) in platelets. This response involved both influx of extracellular Ca(2+) and release of Ca(2+) from intracellular stores. However, in comparison with other platelet agonists, i.e. thrombin and adenosine 5'-diphosphate (ADP), LPA was a very weak Ca(2+)-elevating agent. Furthermore, we observed that the LPA-induced rise in [Ca(2+)](i) was markedly suppressed by cyclic nucleotide-elevating agents. In functional studies, LPA failed to stimulate platelet aggregation and secretion. However, in combination with adrenaline, another weak platelet agonist, LPA could induce an irreversible and complete aggregatory response. There was an individual variation in aggregatory response and tyrosine phosphorylation when LPA and adrenaline were combined. These agents induced a powerful response on platelets from some individuals, but had a weak or no effect on others. INTERPRETATION AND CONCLUSIONS: The present study shows, for the first time, that isolated platelets from some healthy blood donors respond synergistically to a combination of LPA and adrenaline. Platelet activation is a key step in distinguishing normal hemostasis from pathologic hemostasis. Increased knowledge about this mechanism might help to predict individual responses and provide new insights into molecular mechanisms responsible for pathologic thrombosis. PMID- 12091125 TI - Inflammation, fibrinogen and thrombin generation in patients with previous myocardial infarction. AB - BACKGROUND AND OBJECTIVES: It has been reported that the influence of plasma fibrinogen on the incidence of myocardial infarction is related to inflammatory processes. The aim of this study was to investigate the relationship between inflammatory activity, fibrinogen and thrombin generation in patients 5 years after the acute phase of myocardial infarction. DESIGN AND METHODS: Sixty-seven patients 5 years after a myocardial infarction and 67 control subjects were studied. Plasma fibrinogen protein (Fg-protein) and function (Fg-function), prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complex (TAT), total sialic acid (TSA) and C-reactive protein (CRP) were measured. RESULTS: The levels of Fg-protein, Fg-function, F1+2, TAT, TSA and CRP were significantly higher in patients than in the control subjects. Plasma TSA correlated with CRP (r=0.31, p<0.05). There was a significant correlation between TSA and Fg-protein or Fg function (r=0.48, p<0.01). CRP correlated with Fg-function (r=0.32, p<0.05) while there was no correlation between CRP and Fg-protein. CRP also correlated with F1+2 and TAT (r= 0.4, p<0.01). INTERPRETATION AND CONCLUSIONS: Five years after myocardial infarction there was clear evidence of low-grade inflammation that was accompanied by an increase in thrombin formation. The increase of the plasma fibrinogen level is mainly related to TSA and the increase of CRP, which is associated with thrombin generation. PMID- 12091126 TI - Thrombosis prophylaxis in medical patients: a retrospective review of clinical practice patterns. AB - BACKGROUND AND OBJECTIVES: The risk of venous thromboembolism in medical patients is comparable to the risk in general surgical patients. Thromboprophylaxis is recommended for specific medical patients, but its use in clinical practice is unknown. DESIGN AND METHODS: We conducted a retrospective review of the charts of consecutive patients discharged from 2 departments of Internal Medicine, one in the teaching hospital of Varese and one in the non-teaching hospital of Angera, Italy, from October to December 2000. We selected the charts of patients with clinical conditions at increased risk of venous thromboembolism requiring thromboprophylaxis according to consensus statements. The use of antithrombotic drugs and contraindications to prophylaxis were documented. RESULTS: We screened a total of 516 charts, 265 in Varese and 251 in Angera and we identified 165 patients (103 and 62, respectively) at risk of venous thromboembolism because of malignancy (53), heart failure (34), stroke (33), acute infections (23), acute respiratory failure (18), acute rheumatic disorders (3), and inflammatory bowel disease (1). Forty-two patients had contraindications to antithrombotic drugs and 11 were already on long-term oral anticoagulant treatment. Among the 112 remaining patients, prophylaxis was prescribed to 52 patients (46.4%), 35 of 60 in Varese (58.3%) and 17 of 52 in Angera (32.7%, p=0.0067). Patients with stroke and heart failure were significantly more likely to receive thromboprophylaxis than other groups of patients. INTERPRETATION AND CONCLUSIONS: Prophylaxis of venous thromboembolism is underused in medical patients and the proportion of patients receiving antithrombotic drugs varies with the medical condition which precipitated hospital admission. The low rate of usage of prophylaxis suggests that preventable cases of thromboembolism are occurring and that better education of physicians is required to increase the usage of thromboprophylaxis. PMID- 12091127 TI - Methionine synthase polymorphism A2756G is associated with susceptibility for thromboembolic events and altered B vitamin/thiol metabolism. AB - BACKGROUND AND OBJECTIVES: Vitamin B12 dependent methionine synthase (MS) regulates de novo production of methionine from homocysteine (Hcy). Since moderate elevations in Hcy are considered vasculotoxic, we examined a common variant (A2756G-MS) of the gene coding for this enzyme as a risk for thromboembolism. DESIGN AND METHODS: We investigated A2756G-MS and folate/thiol status in 51 individuals who had experienced a thromboembolic event (TE) and 95 subjects being treated for non-thromboembolic (NTE) vascular problems. RESULTS: The prevalence of the mutant G allele was lower in TE subjects than in controls, indicating a protective role for this base substitution (OR 0.39; 95%CI 0.20 0.78; p=0.010). Consistent with an advantage conferred by this allele, heterozygotes had generally lower levels of Hcy and glutathione (GSH), and higher levels of B-vitamins than wildtypes. The OR for the wildtype having an increased risk for TE was 2.32 (95%CI 1.06-5.08). Additionally, as might be predicted, TE wildtypes had elevated GSH levels compared to corresponding NTE-wildtypes (p=0.004) - a likely response to oxidative stress. NTE subjects showed a dramatic reduction in Hcy between wildtype and heterozygote (p=0.017), and again between recessive and heterozygote genotypes (p=0.002). The same pattern, although not significant, occurred in TE subjects. The similarity in Hcy between clinical groups for each genotype raises questions on the etiological role of Hcy in TE. The functional relationship between enzyme variant and its B12-cofactor may be of more interest, since the polymorphic site occurs near the B12-binding domain, and our results indicate wildtype-TE subjects have a much lower level of vitamin B12 than heterozygote-TE subjects (p=0.0019). This effect is attenuated in NTE subjects. INTERPRETATION AND CONCLUSIONS: . A2756G-MS may protect against a thromboembolic event. The role of Hcy remains unclear. PMID- 12091128 TI - Perspectives on antithrombotic agents: from unfractionated heparin to new antithrombotics. AB - BACKGROUND AND OBJECTIVES: Unfractionated heparin (UFH) has been the antithrombotic agent of choice for the prevention and treatment of venous thromboembolism (VTE) for a long time. UFH is also widely used for the treatment of patients with acute coronary syndromes. However, UFH has some limitations such as the need for parenteral administration and close monitoring of its anticoagulant effect. UFH is also associated with bleeding, heparin-induced thrombocytopenia and osteoporosis. EVIDENCE AND INFORMATION SOURCES: Low molecular weight heparins (LMWHs) are produced by the depolymerization of UFH. LMWHs have pharmacologic advantages over UFH: a better bioavailability after subcutaneous administration, a longer plasma half-life and a more predictable anticoagulant effect. These improved features allow once or twice daily subcutaneous injection of weight-adjusted doses of LMWHs without requiring laboratory monitoring in patients with VTE or unstable angina. PERSPECTIVES: A number of new antithrombotic agents are currently under development. These include direct antithrombins and factor Xa inhibitors. The results of the main clinical trials with LMWHs as well as those of the studies with the new antithrombotic agents will be reviewed in this article. PMID- 12091129 TI - Avascular necrosis of the femoral head among children and adolescents with sickle cell disease in Greece. AB - Hemoglobinopathies are very common in Greece, the incidence of beta-thalassemia trait being 8% and that of sickle cell trait ranging from 1 to 32% in various districts. In Greek populations, sickle cell disease (SCD) is mainly represented by S-beta thalassemia. PMID- 12091130 TI - HLA-DRB1*15 and pediatric aplastic anemia. AB - We report a positive association between HLA-DRB1*15 (p= 0.0002) in Turkish patients with pediatric severe aplastic anemia (SAA) and a paradoxically favorable influence of the susceptibility marker on the clinical response to immunosuppressive therapy. These findings point to an immune mechanism mediated by DRB1*15 in SAA which confers responsiveness to treatment. PMID- 12091132 TI - T(15;17) in acute promyelocytic leukemia is not associated with submicroscopic deletions on der(17). AB - We report a fluorescent in situ hybridization (FISH) study on 34 patients with acute promyelocytic leukemia. The study was designed to detect microdeletions in the derivative chromosome 17 which is the result of a reciprocal translocation t(15;17). No deletion was found. PMID- 12091131 TI - Elevated serum vascular endothelial growth factor levels in patients with polycythemia vera and thrombotic complications. AB - Vascular endothelial growth factor (VEGF) induces platelet activation in a thrombin-dependent manner. We tested the serum VEGF levels in patients with polycythemia vera (PV) and found a significant correlation between increased VEGF and thrombosis. These findings suggest that high VEGF levels might contribute to the occurrence of thrombosis in this hematologic malignancy. PMID- 12091133 TI - A new translocation t(11;13)(q13;q14) in a mature B-cell neoplasm. AB - We present the case of a man affected by an unclassified mature B-cell neoplasm with a bone marrow culture stimulated with TPA showing a 46,XY, t(11;13)(q13;q14)[14]/46,XY [6] karyotype. Fluorescent in situ hybridization demonstrated that the BCL1 oncogene is translocated (not rearranged) to chromosome band 13q14 and that a copy of D13S319 locus is deleted. To our knowledge, this is the first reported case with this novel cytogenetic aberration. PMID- 12091134 TI - Early chemosensitivity to VAD regimen predicts a favorable outcome after autologous stem cell transplantation in multiple myeloma. AB - We report that early chemosensitivity, defined by a greater than 50% reduction of M-component and plasma-cell marrow infiltration, after 2 cycles of VAD was correlated with a favorable outcome following autologous stem cell transplantation in 46 patients with newly diagnosed multiple myeloma submitted to high-dose therapy. PMID- 12091135 TI - Mixed chimerism after bone marrow transplantation for thalassemia major. AB - Thirty-four thalassemia patients were studied for chimerism by fluorescent in situ hybridization or variable number tandem repeats after bone marrow transplantation. Mixed chimerism was detected in 9 patients with host cells ranging from 4 to 56%. One had graft rejection and the others were transfusion independent. Mixed chimerism was common but mostly without deleterious effect. PMID- 12091136 TI - Increasing risk of relapse after allogeneic stem cell transplant for adult acute lymphoblastic leukemia in > or = 2nd complete remission induced by highly intensive chemotherapy. AB - From this retrospective single center analysis adults with acute lymphoblastic leukemia transplanted in > or 2nd CR from an HLA-identical sibling later than 1993 had a worse outcome. As the transplanted-related mortality improved by time,this result was essentially due to the increased relapse rate. The intensity of the pre-transplant salvage chemotherapy was identified as the main factor influencing the post-transplant relapse-risk. PMID- 12091137 TI - A severe adverse event in an elderly patient undergoing preoperative autologous blood donation. PMID- 12091138 TI - Systemic aspergillosis in a patient with non-Hodgkin's lymphoma developing acute graft-versus-host disease after autologous peripheral blood stem cell transplantation. AB - Graft-versus-host disease (GVHD) is rare in the autologous setting. We describe a non-Hodgkin's lymphoma case developing acute GVHD after autologous peripheral blood stem cell transplantation following several lines of chemotherapy inclusive of fludarabine. At day +33, he complained of fever, diffused erythematous papulosis with ulceration of skin lesions. A punch biopsy indicated a grade III GVHD. A dose escalation of corticosteroids, cyclosporin-A and photoapheresis induced a transient response. He developed positivity to CMV and systemic aspergillosis. He died at day +185 in haematological complete remission, despite infection-oriented treatment. In spite of the use of prophylactic immunosuppressive drugs, between 50% and 70% of patients given HLA-identical marrow graft develop acute graft-versus-host disease (GVHD) that, in turn, significantly increases the risk of transplant-related mortality. Autologous BMT has been shown to be an effective procedure in several malignancies, persistently becoming a first-line choice in treating patients affected with lymphoproliferative disorders, specially non-Hodgkin's lymphoma (NHL). Although GVHD is a very rare event in the autologous setting (AuGVHD), a consistent number of reports dealing with GVHD-like phenomena has emerged, especially in breast cancer patients. More often, AuGVHD has been induced by the use of immunosuppressive agents, such as cyclosporin-A (CSA), in attempt to evoke a graft-versus-tumor (GVT) effect. However, AuGVHD is mild and self-limited phenomenon. We report the case of a NHL patient who developed unresponsive GVHD after autologous peripheral blood stem cell transplantation (PBSCT). Because of the immunosuppressive therapies, he developed systemic aspergillosis. He died in haematological complete remission despite infection-oriented treatment. PMID- 12091139 TI - Successful treatment of Dieulafoy's disease in a von Willebrand patient. PMID- 12091142 TI - Thymoma and pancytopenia: a very rare association. PMID- 12091141 TI - Progressive multifocal leukoencephalopathy shortly after the diagnosis of follicular lymphoma in a patient treated with fludarabine. PMID- 12091140 TI - Expansion of cells with trisomy 12 associated with progressive lymphocytosis and a dramatic change in chimera status in a patient with chronic lymphocytic leukemia. PMID- 12091143 TI - An unusual clinical course following deep venous thrombosis. PMID- 12091144 TI - An unresolved hematoma. PMID- 12091145 TI - Transcription factor GATA-4 is not involved in Diamond-Blackfan anaemia. PMID- 12091146 TI - Addendum to strategies to deter blood doping in sports. PMID- 12091148 TI - Inefficacy of rituximab in a case of low grade non-Hodgkin's lymphoma with cryoglobulinemia. PMID- 12091149 TI - Changes in whole-tree water relations during ontogeny of Pinus flexilis and Pinus ponderosa in a high-elevation meadow. AB - We measured sap flux in Pinus ponderosa Laws. and Pinus flexilis James trees in a high-elevation meadow in northern Arizona that has been invaded by conifers over the last 150 years. Sap flux and environmental data were collected from July 1 to September 1, 2000, and used to estimate leaf specific transpiration rate (El), canopy conductance (Gc) and whole-plant hydraulic conductance (Kh). Leaf area to sapwood area ratio (LA/SA) increased with increasing tree size in P. flexilis, but decreased with increasing tree size in P. ponderosa. Both Gc and Kh decreased with increasing tree size in P. flexilis, and showed no clear trends with tree size in P. ponderosa. For both species, Gc was lower in the summer dry season than in the summer rainy season, but El did not change between wet and dry summer seasons. Midday water potential (Psi(mid)) did not change across seasons for either species, whereas predawn water potential (Psi(pre)) tracked variation in soil water content across seasons. Pinus flexilis showed greater stomatal response to vapor pressure deficit (VPD) and maintained higher Psi(mid) than P. ponderosa. Both species showed greater sensitivity to VPD at high photosynthetically active radiation (PAR; > 2500 micromol m-2 s-1) than at low PAR (< 2500 micromol m-2 s-1). We conclude that the direction of change in Gc and Kh with increasing tree size differed between co-occurring Pinus species, and was influenced by changes in LA/SA. Whole-tree water use and El were similar between wet and dry summer seasons, possibly because of tight stomatal control over water loss. PMID- 12091150 TI - Photosynthetic responses to water deficit in six Mediterranean sclerophyll species: possible factors explaining the declining distribution of Rhamnus ludovici-salvatoris, an endemic Balearic species. AB - We sought to explain the declining distribution in the Balearic Islands of the endemic shrub Rhamnus ludovici-salvatoris R. Chodat, by comparing its photosynthetic response to drought with that of several widely distributed, competing Mediterranean species (R. alaternus L., Quercus ilex L., Pistacia lentiscus L., Q. humilis Mill. and P. terebinthus L.). All of the study species, except for the two Rhamnus species, avoided desiccation by rapidly adjusting their stomatal conductance at the onset of drought, and maintaining constant leaf relative water content. The two Rhamnus species showed desiccation-tolerant behavior; i.e., as drought progressed, their predawn leaf relative water content decreased simultaneously with stomatal closure. All four desiccation-avoiding species showed a significant positive correlation between leaf thermal dissipation (estimated by the fluorescence parameter NPQ (non-photochemical quenching)) and the de-epoxidation state of the xanthophyll cycle (DPS). The two Rhamnus species exhibited maximum DPS regardless of treatment, but only R. alaternus increased NPQ in response to drought. Rhamnus ludovici-salvatoris had a high ratio of photorespiration to photosynthesis and a low intrinsic water-use efficiency; traits that are likely to be unfavorable for plant productivity under arid conditions. It also had the lowest DPS and thermal dissipation among the six species. We conclude that the photosynthetic traits of R. ludovici-salvatoris account for its limited ability to compete with other species in the Mediterranean region. PMID- 12091151 TI - Gas exchange, biomass, whole-plant water-use efficiency and water uptake of peach (Prunus persica) seedlings in response to elevated carbon dioxide concentration and water availability. AB - We examined the interactive effects of elevated CO2 concentration ([CO2]) and water stress on growth and physiology of 1-year-old peach (Prunus persica L.) seedlings grown in 10-dm3 pots in open-top chambers with ambient (350 micromol mol-1) or elevated (700 micromol mol-1) [CO2]. Seedlings were supplied weekly with a non-limiting nutrient solution. Water was withheld from half of the plants in each treatment for a 4-week drying cycle, to simulate a sudden and severe water stress during the phase of rapid plant growth. Throughout the growing season, seedlings in elevated [CO2] had higher assimilation rates, measured at the growth [CO2], than seedlings in ambient [CO2], and this caused an increase in total dry mass of about 33%. Stomatal conductance, total water uptake, leaf area and leaf number were unaffected by elevated [CO2]. Because seedlings in the two CO2 treatments had similar transpiration despite large differences in total dry mass, water-use efficiency (WUE) of well-watered and water-stressed seedlings grown in elevated [CO2] was an average of 51 and 63% higher, respectively, than WUE of comparable seedlings grown in ambient [CO2]. Elevated [CO2] enhanced total biomass of water-stressed seedlings by 31%, and thus ameliorated the effects of water limitation. However, the percentage increases in total dry mass between well-watered and water-stressed seedlings were similar in ambient (53%) and elevated (58%) [CO2], demonstrating that there was no interaction between elevated [CO2] and water stress. This finding should be considered when predicting responses of trees to global climate change in hot and dry environments, where predicted temperature increases will raise evaporative demands and exacerbate the effects of drought on tree growth. PMID- 12091153 TI - Partitioning of nitrate assimilation among leaves, stems and roots of poplar. AB - Plants differ in tissue localization of nitrate reduction and assimilation. Some species reduce nitrate primarily in the leaves, whereas other species localize nitrate reduction and assimilation in the roots. We determined how nitrate assimilation is partitioned among leaves, stems and roots of poplar (Populus tremula L. x P. alba L.) by comparing tissue differences in in vivo nitrate reductase activity (NRA), nitrate reductase abundance and tissue nitrate concentration. Compared with stems or roots, NRA was greater in leaves, and the highest leaf NRA was found in young leaves. Leaf and root NRA increased with increasing nitrate supply, whereas stem NRA remained constant. Leaf NRA was at least 10-fold greater than root NRA at all external nitrate concentrations. Nitrate reductase abundance increased in all tissues with increasing nitrate availability, and nitrate reductase abundance was at least 10-fold greater in leaves than in stems or roots at all nitrate availabilities. Tissue nitrate concentration increased with increasing nitrate supply and was greater in roots than in stems and leaves. Photoperiod influenced NRA, with leaf NRA declining in nitrate-fertilized plants with short daily photoperiods (8-h). We conclude that different tissues of poplar vary in nitrate assimilation with little nitrate assimilation occurring in roots and the most nitrate assimilation taking place in leaves. PMID- 12091154 TI - Coarse and fine root respiration in aspen (Populus tremuloides). AB - Coarse and fine root respiration rates of aspen (Populus tremuloides Michx.) were measured at 5, 15 and 25 degrees C. Coarse roots ranged from 0.65 to 4.45 cm in diameter, whereas fine roots were less than 5 mm in diameter. To discriminate between maintenance and growth respiration, root respiration rates were measured during aboveground growing periods and dormant periods. An additional measurement of coarse root respiration was made during spring leaf flush, to evaluate the effect of mobilization of resources for leaf expansion on root respiration. Fine roots respired at much higher rates than coarse roots, with a mean rate at 15 degrees C of 1290 micromol CO2 m-3 s-1 during the growing period, and 660 micromol CO2 m-3 s-1 during the dormant period. The temperature response of fine root respiration rate was nonlinear: mean Q10 was 3.90 for measurements made at 5 15 degrees C and 2.19 for measurements made at 15-25 degrees C. Coarse root respiration rates measured at 15 degrees C in late fall (dormant season) were higher (370 micromol CO2 m-3 s-1) than rates from roots collected at leaf flush and early summer (200 micromol CO2 m-3 s-1). The higher respiration rates in late fall, which were accompanied by decreased total nonstructural carbohydrate (TNC) concentrations, suggest that respiration rates in late fall included growth expenditures, reflecting recent radial growth. Neither bud flush nor shoot growth of the trees caused an increase in coarse root respiration or a decrease in TNC concentrations, suggesting a limited role of coarse roots as reserve storage organs for spring shoot growth, and a lack of synchronization between above- and belowground growth. Pooling the data from the coarse and fine roots showed a positive correlation between nitrogen concentration and respiration rate. PMID- 12091152 TI - Needle life span, photosynthetic rate and nutrient concentration of Picea glehnii, P. jezoensis and P. abies planted on serpentine soil in northern Japan. AB - We investigated the adaptation of three spruce species (Picea glehnii Masters, P. jezoensis Carr. and P. abies Karst.) to growth in northern Japan on serpentine soils (characterized by high concentrations of heavy metals and Mg, a low Ca/Mg ratio and low fertility) and fertile brown forest soils. Among species, seedling survival on serpentine soil was highest in P. glehnii. Shoot growth of P. glehnii was similar whether grown on serpentine or brown forest soil, whereas shoot growth of the other species was significantly less on serpentine soil than on brown forest soil. On serpentine soil, needle life span of P. glehnii was at least 3 years longer than that of the other two species. Needle area per shoot of P. glehnii was significantly higher on serpentine soil than on brown forest soil up to a shoot age of 8 years. In all three species, light-saturated photosynthetic rate (Pmax) decreased with needle age independently of soil type. However, on serpentine soil, Pmax in P. glehnii was higher, particularly in older needles, than in the other species. Furthermore, on serpentine soil, needle concentrations of nitrogen and phosphorus were higher in P. glehnii than in the other species. We conclude that P. glehnii is better adapted to serpentine soil than P. jezoensis and P. abies at least in part because of its greater needle life span and higher needle nutrient concentrations. PMID- 12091155 TI - Effects of phenology, water availability and seed source on loblolly pine biomass partitioning and transpiration. AB - One-year-old loblolly pine (Pinus taeda L.) seedlings from four seed sources (Arkansas, Georgia, Texas and Virginia) grown in 1-m-deep sand-filled pits in two water regimes (well-watered and drought) were studied, to gain insight into the process of seedling establishment. Whole-plant transpiration was measured biweekly from July to December. Whole-plant harvests were conducted at 6-week intervals from April to December. Whole-plant transpiration and transpiration per unit leaf and root area were affected by treatment, seedlot and phenology. Seedlings of the Arkansas seedlot maintained significantly higher transpiration rates per unit leaf and root area during drought than seedlings of the Virginia, Georgia or Texas seedlots, but did not accumulate greater biomass. The high transpiration rates of the Arkansas seedlings were attributed to their deep root systems. Allometric relationships indicated that, relative to the whole plant, biomass allocation to needles of drought-treated seedlings was enhanced during the summer (allometric ratio 1.09), whereas allocation to roots was enhanced in the spring and fall (allometric ratios of 1.13 and 1.09, respectively). Relative to the whole plant, biomass allocation to needles of well-watered seedlings was enhanced throughout the experiment (allometric ratio of 1.16 declining to 1.05), whereas the allometric ratio of root to total biomass was 0.89 or less throughout. Allometric relationships also indicated variation in biomass partitioning to roots in three soil layers (0-30, 30-60 and 60-100 cm), which differed among harvests in each soil layer. Root growth in both well-watered and drought-treated seedlings was concentrated in the top soil layer in the spring, shifted to the middle and bottom soil layers in the summer, and then increased in the top soil layer in the fall. Compared with well-watered seedlings, drought treated seedlings had higher rates of root growth in the bottom soil layer in the fall, a characteristic that would confer tolerance to future periods of limited soil water availability. PMID- 12091156 TI - Carbohydrates in individual poplar fine roots: effects of root age and defoliation. AB - Late-summer starch accumulation in fine roots of poplars (Populus x canadensis Moench.) defoliated by gypsy moth (Lymantria dispar L.) lagged behind that in fine roots of undefoliated trees. If starch concentration declines with age, defoliation-induced changes in root system age structure could be partly responsible for this difference. To test this hypothesis, we measured fine-root starch and soluble sugar concentrations in roots of known age from trees in defoliated and undefoliated plots. There was a significant interaction between the effects of defoliation and root type (white, brown, or dead) on fine root soluble sugar concentration because of the high concentration of soluble sugars in white roots from trees in undefoliated plots. Both root starch and soluble sugar concentrations were variable among individuals of each age class. The population frequency distributions for starch and soluble sugar concentrations were both right-skewed, and fit by exponential functions. These data are most consistent with direct defoliation effects on a labile and dynamic pool of carbohydrates in poplar fine roots, rather than indirect defoliation effects on root system age structure. PMID- 12091157 TI - The microcirculation unveiled. PMID- 12091158 TI - When ventilator and patient's end of inspiration don't coincide: what's the matter? PMID- 12091160 TI - Pro: mice are a good model of human airway disease. PMID- 12091159 TI - Studies of the early bactericidal activity of new drugs for tuberculosis: a help or a hindrance to antituberculosis drug development? PMID- 12091161 TI - Con: mice are not a good model of human airway disease. PMID- 12091164 TI - Update in clinical toxicology. PMID- 12091165 TI - Adenosine deaminase inhibition attenuates microvascular dysfunction and improves survival in sepsis. AB - The ability of increased endogenous adenosine to mitigate microvascular derangements in sepsis was studied. Pentostatin (2'-deoxycoformycin), an inhibitor of adenosine deaminase, was administered to mice immediately after induction of sepsis by cecal ligation and puncture. Intravital video microscopy of cremasteric postcapillary venules was performed. Leukocyte rolling and adhesion were significantly increased in septic mice compared with control mice. Treatment of septic mice with pentostatin significantly decreased leukocyte rolling and adhesion (6.02 +/- 0.09 versus 1.72 +/- 0.12 rolling cells/min, 2.07 +/- 0.04 versus 0.62 +/- 0.05 adherent cells/100 microm per minute; p < 0.001). Albumin leakage (ratio) was significantly attenuated in septic animals treated with pentostatin (0.42 +/- 0.05 versus 0.21 +/- 0.04; p < 0.01). Circulating levels of interleukin-6, tumor necrosis factor-alpha, and soluble tumor necrosis factor type II receptor were decreased in septic mice treated with pentostatin. Survival was significantly improved at 48 hours in mice treated with pentostatin. These results suggest an important role for adenosine in modulating both leukocyte-dependent and -independent mechanisms of endothelial injury in sepsis. Exploiting the advantageous action of endogenous adenosine represents a potentially useful and novel therapeutic approach for the treatment of sepsis. PMID- 12091166 TI - Response of ventilator-dependent patients to delayed opening of exhalation valve. AB - In conventional mechanical ventilation, the inflation cycle often extends into neural expiration (TEN), potentially exacerbating dynamic hyperinflation (DH). We wished to determine the extent to which patients defend against DH when this happens. Such defense may include prolongation of TEN (timing response) and/or expiratory muscle recruitment (neuromuscular response). Fifty patients were ventilated in the Proportional Assist mode, allowing us to infer these responses noninvasively. At random intervals, exhalation of single breaths was delayed by briefly delaying the opening of exhalation valve (occlusion) (T(occ) = 0.78 +/- 0.34 seconds). Timing response was assessed from the change in TEN. Neuromuscular response was assessed from the difference between volume exhaled after release of occlusion and volume exhaled in unoccluded breaths over a similar expiratory flow duration (DeltaV(iso-time)). There was no evidence of an acute neuromuscular response; DeltaV(iso-time) averaged 0.005 +/- 0.023 L (NS). Forty-five of 50 patients significantly lengthened TEN. However, the timing response offset only 36 +/- 20% of the delay in expiration. Because of absent neuromuscular responses and weak timing responses, DH increased in most patients in postocclusion breaths (DeltaDH = 0.10 +/- 0.08 L, p = 2E-10). We conclude that acute compensatory responses to delays in opening of exhalation value are weak in ventilator dependent patients. As a result, such nonsynchrony tends to exacerbate DH. PMID- 12091167 TI - A reiterative method for calculating the early bactericidal activity of antituberculosis drugs. AB - Studies of early bactericidal activity (EBA) are important in the rapid evaluation of new antituberculosis drugs. Historically, these have concentrated on the log fall in the viable count in sputum during the first 48 hours of therapy. In this paper, we provide a mathematical model that suggests that the viable count in sputum follows an exponential decay curve with the equation V = S + Me(-kt) (where V is the viable count, M the population of bacteria susceptible to the test drug, S the population susceptible only to sterilizing agents, t the day of sputum collection as related to start of therapy, k the rate constant for the bacteria killed each day, and e the Napierian constant). We demonstrate that data from clinical trials fits the exponential decay model. We propose that future EBA studies should be performed by measuring daily quantitative counts for at least 5 days. We also propose that the comparison of the early bactericidal activity of antituberculosis drugs should be evaluated using the time taken to reduce the viable count by 50% (vt(50)). A further reiterative refinement following a rule set based on statistically the best fit to the exponential decay model is described that will allow investigators to identify anomalous results and thus enhance the accuracy in measuring early bactericidal activity. PMID- 12091168 TI - Transmission dynamics of tuberculosis in Tarrant county, Texas. AB - To understand the transmission dynamics of tuberculosis in Tarrant County, Texas, we performed a population-based study of 159 patients with culture-proven tuberculosis, combining restriction fragment length polymorphism (RFLP) analysis of Mycobacterium tuberculosis isolates with prospective interviewing to identify epidemiologic links between patients. Patients whose isolates had identical or closely related RFLP patterns were considered a cluster. Seventy-six (48%) of 159 patients were in 19 clusters, suggesting that recent transmission accounted for 36% of tuberculosis morbidity. Unconditional logistic regression showed that birth in the United States, continuous residence in Tarrant County, a history of homelessness, and a history of visiting or working in bars were independent predictors of clustering. Four homeless shelters and five bars were associated with specific clusters, suggesting that they were sites of tuberculosis transmission. Patients in some clusters recognized more photographs of patients in their cluster than did patients outside their cluster. We conclude that (1) homeless shelters and bars are important sites of tuberculosis transmission in Tarrant County, and (2) the use of photograph recognition of patients with tuberculosis, in combination with RFLP analysis, has the potential to enhance tuberculosis control by facilitating identification of epidemiologic links between patients. PMID- 12091169 TI - Association between TNFA polymorphism and the development of asthma in the Japanese population. AB - Tumor necrosis factor (TNF) is a proinflammatory cytokine that participates in the inflammatory reaction in patients with asthma. The TNFA and TNFB genes, which encode TNF-alpha and TNF-beta, respectively, are located within the region encoding the human major histocompatibility complex on chromosome 6p21.3, which showed linkage to atopic asthma in our genome-wide search. To determine whether polymorphisms in the 5' flanking region of the TNFA gene (-1031C/T, -863C/A, and 857C/T) and an NcoI polymorphism in the TNFB gene (LTA NcoI) are associated with the development of asthma, we performed transmission disequilibrium tests of families identified through children with atopic asthma. Genotypes of families were determined by polymerase chain reaction-based restriction fragment length polymorphism or SNaPshot analysis. Transmission disequilibrium tests of 144 asthmatic families revealed that transmission of the -857C allele and the -1031T 863C-857C haplotype in the TNFA gene to asthma-affected offspring occurred more frequently than expected (-857C allele, p = 0.0055; -1031T-863C-857C haplotype, p = 0.0002). Our results suggest that TNFA or nearby genes, including those in the major histocompatibility complex region, may contribute to the development of asthma in the Japanese population. PMID- 12091170 TI - P3-specific amplitude reductions to respiratory and auditory stimuli in subjects with asthma. AB - The respiratory-related evoked potential (RREP) is the electroencephalographic response to brief inspiratory occlusion. The P3 component of the response reflects the active cognitive processing of stimulus information. This study investigated the RREP in 16 subjects with asthma and 16 control subjects. In addition, auditory-evoked potentials were recorded using an oddball paradigm in both groups and were compared with the RREP results. Perceptual sensitivity was assessed using a load magnitude estimation task, whereby subjects estimated the magnitude of four added resistive loads using a modified Borg scale. For RREP assessment, inspiratory occlusions were presented for 200 milliseconds. For auditory-evoked potential assessment, 1,000- and 2,000-Hz pure tones were presented at 60 dB for 100 milliseconds, with the 2,000-Hz tone presented with 20% probability and identified as a target stimulus. Scalp electroencephalographic activity was measured from 29 channels. Resistive load intensity was linearly related to magnitude estimation in both subjects with asthma (r2 = 0.96) and control (r2 = 0.99) subjects. Results showed that both the respiratory and auditory P3 components were markedly reduced in the group with asthma compared with the control group. Other components were similar between the groups. These results suggest an intrinsic reduction in P3 amplitude for patients with asthma, which may relate to differences in processing perceptual information. PMID- 12091171 TI - Very late antigen-4 in CD18-independent neutrophil emigration during acute bacterial pneumonia in mice. AB - This study tested the hypothesis that very late antigen (VLA)-4 mediates CD18 independent neutrophil emigration into the airspaces induced by either Streptococcus pneumoniae, a stimulus that induces primarily CD18-independent neutrophil emigration, or Escherichia coli, toward which only 20-30% of the total number of neutrophils emigrate through CD18-independent pathways. In wild-type (WT) mice, VLA-4 expression was less on neutrophils that emigrated into the airspaces than on circulating neutrophils. Vascular cell adhesion molecule-1 (VCAM-1) mRNA, the major endothelial cell ligand for VLA-4, increased more in E. coli than in S. pneumoniae pneumonia. VCAM-1 protein expression was not detected in capillaries, the major site of neutrophil emigration. Neutrophil emigration during E. coli or S. pneumoniae pneumonia was similar in mice given antibodies against both CD18 and VLA-4 compared with mice given the anti-CD18 antibody and a control antibody. However, in hematopoietically reconstituted mice with both WT and CD18-deficient neutrophils in their blood, the migration of CD18-deficient neutrophils in response to S. pneumoniae was slightly but significantly less in animals pretreated with the anti-VLA-4 antibody than in those receiving a control antibody. These data suggest that VLA-4 plays a small role in CD18-independent neutrophil emigration, but the majority of CD18-independent neutrophil emigration induced by bacteria in the lungs occurs through VLA-4-independent mechanisms. PMID- 12091172 TI - Changes in pulmonary mechanics with increasing disease severity in children and young adults with cystic fibrosis. AB - As forced expiratory volume in 1 second (FEV(1)) is a major predictor of outcome in patients with cystic fibrosis (CF), we investigated the effect of FEV(1) on pulmonary mechanics in children and young adults with CF. We measured respiratory rate; tidal volume; minute ventilation; arterial blood gases; sniff esophageal pressure; dynamic lung compliance; total pulmonary resistance; intrinsic positive end expiratory pressure; and total, elastic, and resistive work of breathing in 32 patients (FEV(1) range: 12-49% predicted). We observed correlations between FEV(1) and Pa(O(2)) (r = 0.76, p < 0.0001) and Pa(CO(2)) (r = -0.70, p < 0.0001), FEV(1) and respiratory rate/tidal volume (r = -0.41, p = 0.02), FEV(1) and dynamic lung compliance (r = 0.64, p < 0.0001), and FEV(1) and total work of breathing (r = -0.52, p = 0.002) and elastic work of breathing (r = -0.60. p = 0.0003). No correlations were observed between FEV(1) and sniff esophageal pressure (p = 0.5), minute ventilation (p = 0.9), total pulmonary resistance (p = 0.3), intrinsic positive end expiratory pressure (p = 0.3), or resistive work of breathing (p = 0.1). As FEV(1) declines in children and young adults with CF, there is an increase in the elastic load and work of breathing, resulting in a rapid shallow breathing pattern, that is associated with further impairment of gas exchange. PMID- 12091173 TI - Spirometry in 3- to 6-year-old children with cystic fibrosis. AB - Spirometry is routinely used to assess pulmonary function of older children and adults with cystic fibrosis (CF); however, few data exist concerning the preschool age group. We have reported normative spirometric data for 3- to 6-year old children. The current study was designed to assess a similarly aged group of clinically stable patients with CF. Thirty-three of 38 children with CF were able to perform 2 or 3 technically acceptable maneuvers. These patients had significantly decreased FVC, FEV(1), FEV(1)/FVC, and FEF(25-75) when expressed as z scores (number of SD from predicted): -0.75 +/- 1.63, -1.23 +/- 1.97, -0.87 +/- 1.33, and -0.74 +/- 1.63, respectively. There were significant positive correlations of the Brasfield radiological score with FVC and FEV(1) z scores (r(2) = 0.26, p < 0.01 and r(2) = 0.24, p < 0.01). In addition, homozygous patients for the DeltaF508 mutation had lower z scores for FVC (-1.21 versus 0.47, p < 0.01) and FEV(1) (-1.38 versus 0.21, p < 0.05) than heterozygous patients. Of the 14 patients who had full flow-volume spirometric measurements during infancy, 10 had FEF(25-75) z scores greater than -2 at both evaluations. Our findings suggest that spirometry can successfully be used to assess lung function in preschool children with CF and has the potential for longitudinal assessment from infancy through adulthood. PMID- 12091174 TI - Lack of association between antibiotic use in the first year of life and asthma, allergic rhinitis, or eczema at age 5 years. AB - Five retrospective studies have reported an association between antibiotic use in early life and asthma in childhood. We studied the relationship between the use of oral antibiotics in the first year of life and asthma, allergic rhinitis, and eczema at age 5 years among 448 children with a parental history of atopy monitored from birth. After adjustment for potential confounders, we found no significant association between antibiotic use in the first year of life and asthma (odds ratio [OR] for one versus no courses of antibiotics, 0.5; 95% confidence interval [CI] for OR, 0.2 to 1.5; OR for two or more versus no courses of antibiotics, 1.0; 95% CI for OR, 0.5 to 2.2), recurrent wheezing, allergic rhinitis, or eczema at age 5 years. There was no significant association between antibiotic use in the first year of life and having at least one of three atopic diseases (asthma, allergic rhinitis, or eczema) at age 5 years (OR for one versus no courses of antibiotics, 0.7; 95% CI, 0.4 to 1.4; OR for two or more versus no courses of antibiotics, 0.9; 95% CI, 0.5 to 1.4). Our findings do not support the hypothesis that antibiotic use in early life is associated with the subsequent development of asthma and atopy in childhood. PMID- 12091175 TI - Association between air pollution and lung function growth in southern California children: results from a second cohort. AB - A cohort of 1,678 Southern California children, enrolled as fourth graders in 1996, was followed for 4 years to determine whether the growth in lung function of the children was associated with their exposure to ambient air pollutants. These subjects comprised the second cohort of fourth grade children participating in the Children's Health Study. Significant deficits in lung function growth rate were associated with exposure to acid vapor, NO(2), particles with aerodynamic diameter less than 2.5 microm (PM(2.5)), and elemental carbon. For example, the average annual growth rates of maximal midexpiratory flow and forced expiratory volume in 1 second were reduced by approximately 11% (p = 0.005) and 5% (p = 0.03), respectively, across the observed range of acid exposure. Exposure to acid vapor was also associated with reductions in the ratio of maximal midexpiratory flow to forced vital capacity (p = 0.02), whereas exposure to ozone was correlated with reduced growth in peak flow rate (p = 0.006). Larger deficits in lung function growth rate were observed in children who reported spending more time outdoors. These findings provide important replication of our previous findings of an effect of air pollution on lung function growth that were based on the first fourth-grade cohort from the Children's Health Study (Am J Respir Crit Care Med 2000;162:1383-1390). PMID- 12091176 TI - Laryngeal muscle activities with cerebral hypoxia-ischemia in newborn lambs. AB - This study tested the hypotheses that (1) acute cerebral hypoxia-ischemia changes laryngeal adductor, laryngeal abductor, and diaphragmatic activities, resulting in central apnea with laryngeal closure; and (2) these laryngeal muscle activities act to maintain absolute lung volume. The respiratory pattern was determined in three asphyxiated, awake preterm lambs after cesarean section birth and in 12 awake, term lambs, with normal lung function, after induction of acute cerebral hypoxia-ischemia by occlusion of the brachiocephalic artery. Electrocorticogram activity, flow, volume, electromyograms of laryngeal abductor and adductor muscles and diaphragm, and, in the term lambs, trans-upper airway pressure and carotid blood flow were recorded. With either preterm birth asphyxia or induced acute cerebral hypoxia-ischemia, minute ventilation initially increased, and then hypopnea occurred. During the hypopnea, laryngeal adductor activity was prominent, accompanied by an increased upper airway pressure and a maintained/raised absolute lung volume. Thus, when acute hypoxia-ischemia limited to the upper body is induced in lambs with normal lung function, expiratory laryngeal adduction with closure of the upper airway occurs and likely functions to aid autoresuscitation. PMID- 12091177 TI - Altered breathing pattern after prenatal nicotine exposure in the young lamb. AB - Maternal smoking during pregnancy is a risk factor for sudden fetal and infant death as well as obstructive airway disease in childhood. Fetal nicotine exposure affects organ development. The aim of the present study was to investigate effects of fetal nicotine exposure on lung function in young lambs. Nine unanesthetized, awake, prenatally nicotine-exposed lambs (N) (approximate maternal dose: 0.5 mg/kg) and 12 nonexposed control lambs (C) were studied repeatedly for 5 weeks after birth using a pneumotachograph and a computerized method for breath-by-breath determinations. N and C lambs had similar minute ventilation but a markedly different breathing pattern. At both 5 and 21 days, average age, N lambs had significantly lower tidal volumes and higher respiratory rates than C lambs. Inspiratory drive (P(0.1)) and effective impedance were significantly higher in N lambs compared with C lambs only at 5 days. Prenatal nicotine exposure appears to have long-term effects on the postnatal breathing pattern, suggesting altered lung function, e.g., increased airway resistance, decreased lung compliance, or both. The increased inspiratory drive is most likely secondary to increased impedance of the respiratory system. These changes are most marked close to birth but persist during the initial postnatal period. PMID- 12091178 TI - Microvascular blood flow is altered in patients with sepsis. AB - Microvascular blood flow alterations are frequent in animal models of sepsis and may impair tissue oxygenation. We hypothesized that alterations of the microcirculation are present in patients with sepsis. We used an orthogonal polarization spectral imaging technique to investigate the sublingual microcirculation in 10 healthy volunteers, 16 patients before cardiac surgery, 10 acutely ill patients without sepsis (intensive care unit control subjects), and 50 patients with severe sepsis. The effects of topical application of acetylcholine (10(-2) M) were tested in 11 patients with sepsis. In each subject, five to seven sublingual areas were recorded and analyzed semiquantitatively. Data were analyzed with nonparametric tests and are presented as medians (25th 75th percentiles). No significant difference in microvascular blood flow was observed between healthy volunteers and patients before cardiac surgery or intensive care unit control subjects. The density of all vessels was significantly reduced in patients with severe sepsis (4.5 [4.2-5.2] versus 5.4 [5.4-6.3]/mm in volunteers, p < 0.01). The proportion of perfused small (< 20 microm) vessels was reduced in patients with sepsis (48 [33-61] versus 90 [89 92]% in volunteers, p < 0.001). These alterations were more severe in nonsurvivors. The topical application of acetylcholine totally reversed these alterations. In conclusion, microvascular blood flow alterations are frequent in patients with sepsis and are more severe in patients with a worse outcome. PMID- 12091179 TI - Airway inflammation in severe chronic obstructive pulmonary disease: relationship with lung function and radiologic emphysema. AB - The lung pathology of severe chronic obstructive pulmonary disease (COPD) has been poorly investigated. We examined surgical specimens obtained from patients with severe (forced expiratory volume in 1 second [FEV(1)] = 29 +/- 3% predicted, n = 9) or mild/no airflow limitation (FEV(1) = 86 +/- 5% predicted, n = 9) and similar smoking history. With histochemical and immunohistochemical methods we quantified the structural changes and the inflammatory cells in small airways and in muscular pulmonary arteries. As compared with smokers with mild/no COPD, smokers with severe COPD had an increased number of leukocytes in the small airways, which showed a positive correlation with the radiologic score of emphysema and with the value of residual volume, and a negative correlation with the values of FEV(1) and carbon monoxide diffusing capacity. The inflammatory process was characterized by an increase in CD8(+) and CD4(+) T-lymphocytes in the airway wall and by an increase in macrophages in the airway epithelium. When all smokers were considered together, the smoking history was correlated with both the airway wall and smooth muscle thickness, suggesting that smoking itself may play a role in the development of structural changes. No structural and cellular differences were observed in pulmonary arteries between smokers with severe COPD and smokers with mild/no COPD. In conclusion, in the small airways of smokers with severe COPD, there is an increased number of leukocytes, which is correlated with reduced expiratory flow, lung hyperinflation, carbon monoxide diffusion impairment, and radiologic emphysema, suggesting a role for this inflammatory response in the clinical progression of the disease. PMID- 12091180 TI - ATS statement: guidelines for the six-minute walk test. PMID- 12091181 TI - Mitochondrial disorders and exertional intolerance: controversy continues. PMID- 12091183 TI - Obstetric complications and schizophrenia: historical and meta-analytic review. AB - OBJECTIVE: This paper reviews the literature on obstetric complications as a risk factor for schizophrenia. The authors trace the evolution of this literature through different methods and carry out a quantitative review of the results from prospective, population-based studies. METHOD: Relevant papers were identified by a MEDLINE search, by examination of reference lists of published papers, and through personal contact with researchers in the field. Studies were grouped in chronological order according to common themes or methods. Meta-analytic techniques were used to summarize the findings of prospective population-based studies. RESULTS: The meta-analytic synthesis of the prospective population-based studies revealed that three groups of complications were significantly associated with schizophrenia: 1) complications of pregnancy (bleeding, diabetes, rhesus incompatibility, preeclampsia); 2) abnormal fetal growth and development: (low birthweight, congenital malformations, reduced head circumference), and 3) complications of delivery (uterine atony, asphyxia, emergency Cesarean section). Pooled estimates of effect sizes were generally less than 2. CONCLUSIONS: Current methods of investigating the relationship between obstetric complications and schizophrenia are reaching the limit of their usefulness. Lack of statistical power to measure small and interactive effects and lack of detailed information about the prenatal period are major problems with current approaches. A combination of disciplines and approaches will be needed to elucidate the mechanisms underlying these small but important associations. PMID- 12091184 TI - Slow transcranial magnetic stimulation, long-term depotentiation, and brain hyperexcitability disorders. AB - OBJECTIVE: Many clinical syndromes in neuropsychiatry suggest focal brain activation. Repetitive transcranial magnetic stimulation (rTMS) has been proposed as a method for selectively altering neural activity. METHOD: Studies assessing effects of "slow" rTMS, administered up to once per second, in normal people and in those with pathological conditions are reviewed. The findings are compared with those of animal studies examining long-term depression and long-term depotentiation through direct electrical stimulation of cortical tissue. RESULTS: Data suggest that slow rTMS reduces cortical excitability, both locally and in functionally linked cortical regions. Preliminary studies of patients with focal dystonia, epileptic seizures, and auditory hallucinations indicate symptom reductions following slow rTMS. Long-term depotentiation exhibits many features congruent with those of slow rTMS, including frequency dependence, spread to functionally linked cortical regions, additive efficacy, and extended duration of effects. CONCLUSIONS: Slow rTMS offers a new method for probing and possibly treating brain hyperexcitability syndromes. Further studies linking slow rTMS to animal models of neuroplasticity are indicated. PMID- 12091185 TI - Diseases of the mind and brain: Rett's syndrome. PMID- 12091186 TI - A case of "pfropfschizophrenia": Kraepelin's bridge between neurodegenerative and neurodevelopmental conceptions of schizophrenia. PMID- 12091187 TI - Ernst Kretschmer (1888-1964). PMID- 12091188 TI - Hippocampal changes in patients with a first episode of major depression. AB - OBJECTIVE: Previous work suggests that patients with unipolar depression may have structural as well as functional abnormalities in limbic-thalamic-cortical networks, which are hypothesized to modulate human mood states. A core area in these networks is the hippocampus. In the present study, differences in volumes of hippocampal gray and white matter between patients with a first episode of major depression and healthy comparison subjects were examined. METHOD: Thirty patients with a first episode of major depression and 30 healthy comparison subjects who were matched for age, gender, handedness, and education were examined with high-resolution magnetic resonance imaging. RESULTS: Male patients with a first episode of major depression had significantly smaller hippocampal total and gray matter volumes than healthy male comparison subjects. Both male and female patients showed significant alterations of left-right asymmetry and significant reductions of left and right hippocampal white matter fibers in relation to healthy comparison subjects. Hippocampal measurements were not significantly correlated with clinical variables, such as age at onset of illness, illness duration, or severity of depression. CONCLUSIONS: These results are consistent with findings of structural abnormalities of the hippocampal formation in patients with major depression that were more pronounced in male patients. The authors' findings support the hypothesis that the hippocampus and its connections within limbic-cortical networks may play a crucial role in the pathogenesis of major depression. PMID- 12091189 TI - Executive dysfunction in geriatric depression. AB - OBJECTIVE: The purpose of this study was to characterize the neuropsychological presentation of geriatric depression and to determine whether depression-related executive dysfunction is more pronounced during advanced age. METHOD: The attention and executive functioning of 40 adults with major depression were compared with those of 40 healthy comparison subjects; 20 subjects were 20-60 years old, and 20 were > or =61 years. It was hypothesized that depressed subjects, regardless of age, would perform more poorly than comparison subjects on both attention and executive tasks but that the older depressed adults would evidence significantly greater impairment on executive measures. RESULTS: A significant interaction between age and depressive status was noted for tasks of executive functioning, while no age-depression interaction was found for tasks of selective or sustained attention. Older depressed adults demonstrated the slowest psychomotor speed and the poorest performance on tasks requiring set shifting, problem solving, and initiation of novel responses. CONCLUSIONS: Patients with late-life depression have significant impairment in executive functioning. These findings can guide the development of stimulated functional neuroimaging paradigms that may clarify the pathophysiology of geriatric depression. Timely identification of attentional and executive processes fundamental to the daily functioning of depressed older adults may lead to compensatory strategies that will improve the outcomes of late-life depression. PMID- 12091190 TI - Are depressive symptoms a risk factor for mortality in elderly Japanese American men?: the Honolulu-Asia Aging Study. AB - OBJECTIVE: This study determined the influence of depressive symptoms on subsequent mortality of all causes. METHOD: The Honolulu Heart Program, established in 1965, is a prospective, community-based cohort of Japanese American men living in Hawaii. The analysis was based on 3,196 Japanese American men aged 71-93 at the time of the fourth examination in 1991-1993. Depressive symptoms were measured by use of an 11-question version of the Centers for Epidemiologic Studies Depression Scale questionnaire. All-cause mortality data were available for 6 years of follow up. Data were analyzed on the basis of presence or absence of chronic diseases. RESULTS: The overall prevalence of frequent depressive symptoms was 9.9%. Age-adjusted mortality rates at 3 years were 48.0 and 30.3 per 1,000 person-years for the depressed and nondepressed groups, respectively. At 6 years, the rates were 54.1 (depressed) and 41.5 (nondepressed) per 1,000 person-years. After adjustment for age, marital status, and antidepressant use, the relative risk for all-cause mortality associated with depressive symptoms was 1.53 for 3-year and 1.27 for 6-year mortality. Among participants who were healthy (without cognitive impairment, coronary heart disease, stroke, diabetes, or cancer), the association between depressive symptoms and mortality was greater (relative risk of 2.30 and 1.57 for 3- and 6 year mortality, respectively). Among participants with chronic disease, there were no significant associations between depressive symptoms and mortality. CONCLUSIONS: Depressive symptoms are a risk factor for mortality in elderly people, particularly in physically healthy individuals. PMID- 12091191 TI - Toward a comprehensive developmental model for major depression in women. AB - OBJECTIVE: Major depression is a multifactorial disorder with many etiologic variables that are interrelated through developmental pathways. The authors used structural equation modeling to generate a developmental model for the etiology of major depression in women. METHOD: Data from 1,942 adult female twins, interviewed up to four times over a 9-year period, were used to construct a developmental model to predict depressive episodes in the year before the most recent interview. Eighteen risk factors in five developmental tiers were considered: 1) childhood (genetic risk, disturbed family environment, childhood sexual abuse, and childhood parental loss), 2) early adolescence (neuroticism, self-esteem, and early-onset anxiety and conduct disorder), 3) late adolescence (educational attainment, lifetime traumas, social support, and substance misuse), 4) adulthood (history of divorce and past history of major depression), and 5) the last year (marital problems, difficulties, and stressful life events). RESULTS: The best fitting model included six correlations and 64 paths, provided an excellent fit to the data, and explained 52% of the variance in liability to episodes of major depression. The findings suggest that the development of risk for major depression in women results from three broad pathways reflecting internalizing symptoms, externalizing symptoms, and psychosocial adversity. CONCLUSIONS: Major depression is an etiologically complex disorder, the full understanding of which will require consideration of a broad array of risk factors from multiple domains. These results, while plausible, should be treated with caution because of problems with causal inference, retrospective recall bias, and the limitations of a purely additive statistical model. PMID- 12091192 TI - Combination of a mood stabilizer with risperidone or haloperidol for treatment of acute mania: a double-blind, placebo-controlled comparison of efficacy and safety. AB - OBJECTIVE: The study assessed the efficacy and safety of risperidone as an adjunctive agent to mood stabilizers in the treatment of acute mania. METHOD: This 3-week randomized, double-blind, placebo-controlled study included 156 bipolar disorder patients with a current manic or mixed episode who received a mood stabilizer (lithium or divalproex) and placebo, risperidone, or haloperidol. The primary efficacy measure was the Young Mania Rating Scale. Other assessments used the Brief Psychiatric Rating Scale, the Clinical Global Impression scale, and safety measures. RESULTS: The trial was discontinued by 25 (49%) of the 51 placebo group patients, 18 (35%) of the 52 risperidone group patients, and 28 (53%) of the 53 haloperidol group patients. Mean modal doses were 3.8 mg/day (SD=1.8) of risperidone and 6.2 mg/day (SD=2.9) of haloperidol. Significantly greater reductions in Young Mania Rating Scale scores at endpoint and over time were seen in the risperidone group and in the haloperidol group, compared with the placebo group. Young Mania Rating Scale total scores improved with risperidone and with haloperidol both in patients with psychotic features and in those without psychotic features at baseline. Extrapyramidal Symptom Rating Scale total scores at endpoint were significantly higher in the haloperidol patients than in the placebo patients. Antiparkinsonian medications were received by 8%, 17%, and 38% of patients in the placebo, risperidone, and haloperidol groups, respectively. CONCLUSIONS: Risperidone plus a mood stabilizer was more efficacious than a mood stabilizer alone, and as efficacious as haloperidol plus a mood stabilizer, for the rapid control of manic symptoms and was well tolerated. PMID- 12091193 TI - Effect of abrupt change from standard to low serum levels of lithium: a reanalysis of double-blind lithium maintenance data. AB - OBJECTIVE: Growing evidence suggests that abrupt lithium discontinuation increases the risk of recurrence for patients with bipolar disorder. To assess the effect of abrupt change in lithium dose, the authors reanalyzed data from a previously reported, randomized, double-blind trial of standard- versus low-dose lithium for maintenance therapy in bipolar disorder. METHOD: In the original study, serum lithium levels were obtained during a 2-month open stabilization period for 94 patients with bipolar disorder who were then randomly assigned to be maintained on a low (serum level=0.4-0.6 meq/liter) or a standard (0.8-1.0 meq/liter) level of lithium therapy. Patients were then followed for up to 182 weeks. This reanalysis examined the potential confounding influence of prerandomization lithium level and change in lithium level on the outcome of subjects assigned to a standard or low maintenance dose of lithium. RESULTS: In a Cox proportional hazards model incorporating pre- and postrandomization lithium levels and the interaction of these factors, only the interaction term remained significantly associated with time to recurrence. CONCLUSIONS: The findings indicate that change in serum lithium level may be a more powerful predictor of recurrence of bipolar disorder than the absolute assignment to a low or a standard dose of lithium and suggest that an abrupt decrease in lithium level should be avoided. This reanalysis did not directly address optimal maintenance lithium levels but raises questions about the original study's finding of superiority for lithium levels > or =0.8 meq/liter. The results underscore the importance of accounting for the possible confounding effects of changes in the intensity of pharmacotherapy in studies of maintenance therapies for bipolar disorder. PMID- 12091195 TI - Implications for altered glutamate and GABA metabolism in the dorsolateral prefrontal cortex of aged schizophrenic patients. AB - OBJECTIVE: Pharmacological, clinical, and postmortem studies suggest altered gamma-aminobutyric acid (GABA)-ergic and glutamatergic function in patients with schizophrenia. The dorsolateral prefrontal cortex is one key locus of abnormality. The precise neurochemical mechanisms underlying neurotransmitter alterations, such as hypoglutamatergia or GABA dysfunction, are not well understood. This study investigated key biochemical elements of GABA and glutamate metabolism in brain specimens from schizophrenic patients. The activities of nine principal GABA and glutamate-associated metabolic enzymes were measured concurrently in the dorsolateral prefrontal cortex of antemortem assessed and neuropathologically characterized schizophrenic and comparison subjects. METHOD: Postmortem dorsolateral prefrontal cortex specimens from schizophrenia, Alzheimer's disease, and normal nonpsychiatric comparison subjects were assayed to determine activities of the principal glutamate and GABA metabolizing enzymes glutamine synthetase, glutamate dehydrogenase, alpha ketoglutarate dehydrogenase, phosphate-activated glutaminase, alanine aminotransferase, aspartate aminotransferase, glutamic acid decarboxylase, GABA transaminase, and succinic semialdehyde dehydrogenase. RESULTS: Glutamic acid decarboxylase activities were twofold greater and phosphate-activated glutaminase activities were fourfold greater in the schizophrenic group than in the comparison group. Differences in postmortem interval, tissue pH, inhibition of phosphate-activated glutaminase, and medication effects could not account for the differences. Differences in phosphate-activated glutaminase and glutamic acid decarboxylase activities in equivalent specimens from Alzheimer's patients were not observed. The activities of the remaining enzymes were unchanged. CONCLUSIONS: Greater phosphate-activated glutaminase and glutamic acid decarboxylase activities, specific to schizophrenia patients, provide additional biochemical evidence that dorsolateral prefrontal cortex glutamate and GABA metabolism is altered in schizophrenic subjects. These greater activities are consistent with models of a dysregulated glutamatergic/GABA-ergic state in schizophrenia. PMID- 12091194 TI - Suicide attempts in patients with bipolar I disorder during acute and maintenance phases of intensive treatment with pharmacotherapy and adjunctive psychotherapy. AB - OBJECTIVE: Lifetime rates of suicide attempts among patients with bipolar I disorder were compared to rates during a 2-year period of intensive treatment with pharmacotherapy and with one of two adjunctive psychosocial interventions. METHOD: Subjects entered the study during an acute mood episode. Subjects were treated with primarily lithium pharmacotherapy and with either psychotherapy specific to bipolar disorder, which included help in regularizing daily routines, or nonspecific, intensive clinical management involving regular visits with empathic clinicians. Data on prior suicide attempts were obtained retrospectively from interviews with the NIMH-Life-Chart method. Data on suicide attempts during the clinical trial were collected systematically throughout the protocol. RESULTS: The rate of suicide attempts was 1.05 per 100 person-months before patients entered the trial. Patients experienced a threefold reduction in the rate of suicide attempts during the acute treatment phase (until the patient achieved stabilization, defined by completion of 4 weeks during which the patient had a mean score of < or =7 on the 17-item Hamilton Depression Rating Scale and < or =7 on the Bech-Rafaelsen Mania Scale) and a 17.5-fold reduction during maintenance treatment. Poisson loglinear regression analysis modeling the relationship between the observed rates and the three protocol stages (pretreatment, acute, and maintenance) showed that the reductions were significant in the acute and maintenance phases, compared with the pretreatment phase. No patient with one or more suicide attempts before entering the trial attempted suicide during the protocol. CONCLUSIONS: A treatment program in a maximally supportive clinical environment can significantly reduce suicidal behavior in high-risk patients with bipolar I disorder. PMID- 12091196 TI - Expression of Oct-6, a POU III domain transcription factor, in schizophrenia. AB - OBJECTIVE: Oct-6, a POU-III domain transcription factor, is expressed in embryonic stem cells and Schwann cells and in neuronal subpopulations during early mammalian development, but its relevance to disorders of cerebral development in humans is untested. This study evaluated the expression of Oct-6 in schizophrenia, a disorder that has been linked with neurodevelopmental abnormalities. METHOD: Immunohistochemistry was used to examine Oct-6 expression in the temporal lobe in postmortem tissue from 10 subjects with schizophrenia and 10 matched comparison subjects. Western blot analysis was used to study Oct-6 expression in the frontal and temporal cortex in tissue from an additional three schizophrenic and three matched comparison subjects and in the frontal lobe only in tissue from an additional 10 schizophrenic and 10 matched comparison subjects. RESULTS: Extensive Oct-6 immunoreactivity was present in the temporal lobe in all 10 schizophrenic subjects, while very little or no expression was found in the comparison subjects. In schizophrenic subjects, Oct-6 immunoreactivity was found in a subset of cells in the pyramidal cell layer of the hippocampus and in the granule cell layer of the dentate gyrus. Oct-6 staining was predominantly localized in the cytoplasm. Western blot analysis confirmed the presence of Oct-6 in the frontal and temporal cortex in schizophrenic subjects but not in comparison subjects. CONCLUSIONS: The presence of Oct-6 expression in the schizophrenic subjects but not in the comparison subjects suggests that Oct-6 may provide a marker for the neuropathology associated with schizophrenia. Further, it may provide a clue to the neurodevelopmental basis of the disease and could be a reliable means to examine the developmental brain abnormalities described in this disorder. PMID- 12091197 TI - Longitudinal assessment of premorbid cognitive functioning in patients with schizophrenia through examination of standardized scholastic test performance. AB - OBJECTIVE: Studies suggest the presence of premorbid cognitive impairment in patients with schizophrenia, yet the onset and course of these deficits remain unclear. The purpose of this study was to investigate the longitudinal course of premorbid cognitive functioning in individuals with schizophrenia by using prospective data obtained from scholastic test results. METHOD: Scores from grades 4, 8, and 11 on the Iowa Tests of Basic Skills and the Iowa Tests of Educational Development were obtained for 70 subjects who later developed schizophrenia. The mean percentile rank of the subjects' scores in the areas of vocabulary, reading comprehension, language, mathematics, sources of information, and an overall composite category were compared with state norms. RESULTS: The subjects scored below the 50th percentile for each category in all three grades, but only language, reading, sources of information, and composite scores from grade 11 were significantly below state norms. With regard to longitudinal course, there was a significant linear decrease in language scores over time. Scores from grade 11 were positively correlated with WAIS-R IQ, verbal fluency, and Rey Auditory Verbal Learning scores at first illness episode. Scores from grade 11 were not significantly correlated with age at illness onset or ratings of disorganized, psychotic, or negative symptoms. CONCLUSIONS: Scholastic test scores at grades 4 and 8 were nonsignificantly below average in this group of children who later developed schizophrenia. However, test scores dropped significantly between grades 8 and 11. This corresponds to ages 13-16 years, or the onset of puberty. Poor or declining scholastic performance may be a precursor to the cognitive impairment seen during the first episode of illness. PMID- 12091198 TI - MRI study of caudate nucleus volume and its cognitive correlates in neuroleptic naive patients with schizotypal personality disorder. AB - OBJECTIVE: "Cognitive" circuits anatomically link the frontal lobe to subcortical structures; therefore, pathology in any of the core components of these circuits, such as in the caudate nucleus, may result in neurobehavioral syndromes similar to those of the frontal lobe. Neuroleptic medication, however, affects the size of the caudate nucleus. For this reason, individuals diagnosed with schizotypal personality disorder offer an ideal group for the measurement of the caudate nucleus because they may be genetically related to individuals with schizophrenia but do not require neuroleptic treatment because of their less severe symptoms. METHOD: Magnetic resonance imagining (MRI) scans obtained on a 1.5-T magnet with 1.5-mm contiguous slices were used to measure the caudate nucleus and lateral ventricles in 15 right-handed male subjects with schizotypal personality disorder who had no previous neuroleptic exposure and in 14 normal comparison subjects. Subjects were group matched for parental socioeconomic status, handedness, and gender. RESULTS: First, the authors found significantly lower left and right absolute (13.1%, 13.2%) and relative (9.1%, 9.2%) caudate nucleus volumes in never-medicated subjects with schizotypal personality disorder than in normal subjects. Second, they found significant, inverse correlations between caudate nucleus volume and the severity of perseveration in two distinct working memory tasks in these neuroleptic-naive subjects with schizotypal personality disorder. CONCLUSIONS: These data are consistent with the findings of reduced caudate nucleus volume reported in studies of neuroleptic-naive patients experiencing their first episode of schizophrenia and support the association of intrinsic pathology in the caudate nucleus with abnormalities in working memory in the schizophrenia spectrum. PMID- 12091199 TI - Does the definition of borders of the planum temporale influence the results in schizophrenia? AB - OBJECTIVE: The planum temporale, a highly asymmetric neocortical area of the temporal lobe, has a possible role in schizophrenia. The authors used three different anatomical definitions of the planum temporale to examine the anterior, posterior, and total planum temporale gray matter volumes simultaneously. METHOD: Magnetic resonance imaging was used to examine 30 male schizophrenic patients and 30 healthy male comparison subjects. The total planum temporale was identical in all three anatomical definitions applied to determine the border between the anterior and posterior planum temporale regions. RESULTS: No significant differences between men with and without schizophrenia were detected with regard to planum temporale volumes and asymmetry coefficients for any of the three definitions. CONCLUSIONS: The authors could not prove the hypothesis that the definition of planum temporale borders influences the results concerning possible disturbances of planum temporale asymmetry in patients with schizophrenia. PMID- 12091200 TI - Capacity to provide informed consent for participation in schizophrenia and HIV research. AB - OBJECTIVE: The degree to which people with psychiatric symptoms and cognitive dysfunction can provide informed consent to participate in research is a controversial issue. This study was designed to examine the capacity of subjects with schizophrenia and subjects with HIV to provide informed consent for research participation and to determine the relationships among cognitive dysfunction, psychiatric symptoms, and decisional capacity. METHOD: Twenty-five men and women with a DSM-IV diagnosis of schizophrenia and 25 men and women with HIV were recruited. The groups were compared in terms of neuropsychological functioning, psychiatric symptoms, and ability to provide informed consent to a hypothetical drug trial. RESULTS: Eighty percent of the subjects with schizophrenia and 96% of the HIV-positive subjects demonstrated adequate capacity to consent to the hypothetical drug trial, but subjects in the schizophrenia group had significantly lower scores on two of the four aspects of decisional capacity. For the subjects with schizophrenia, neuropsychological functioning and psychiatric symptoms (e.g., apathy and avolition), but not psychotic symptoms (e.g., hallucinations and delusions), were significantly associated with decisional capacity. CONCLUSIONS: The majority of subjects who are recruited and willing to participate in schizophrenia or HIV research will have adequate capacity to provide consent. Cognitive dysfunction and the symptoms shown to be associated with impaired decisional capacity are not unique to schizophrenia and may occur with many other forms of illness. These findings underscore the importance of considering how decisional capacity will be assessed in all types of research, regardless of the specific condition being studied. PMID- 12091201 TI - Cigarette smoking and panic: the role of neuroticism. AB - OBJECTIVE: The authors sought to determine the relationship of panic attacks, cigarette smoking, and neuroticism. METHOD: Data were drawn from the Midlife Development in the United States Survey, a representative household survey of the adult population of the United States (N=3,032). Multivariate logistic regression analyses were used to determine the association between panic attacks and cigarette smoking and to determine whether neuroticism was an independent predictor of the co-occurrence of cigarette smoking and panic attacks. RESULTS: The majority of individuals with panic attacks had been regular smokers during their lifetime (81.1%), a significantly higher rate than seen among subjects without panic attacks (69.4%). Regular cigarette smoking was associated with a significantly greater risk of current panic attacks. This significant association persisted after analyses controlled for demographic characteristics, depression, and alcohol/substance use disorders but was no longer evident after analyses adjusted for neuroticism. Neuroticism independently predicted the co-occurrence of regular cigarette smoking and panic attacks but did not predict either panic attacks or cigarette smoking in the absence of the other. CONCLUSIONS: Consistent with previous findings, these data suggest that panic attacks are associated with greater risk of cigarette smoking but provide new evidence that neuroticism may play an essential role in this relationship. These data are preliminary but, if replicated, suggest that neuroticism may reflect a shared vulnerability for the co-occurrence of cigarette smoking and panic attacks. Future studies are needed to further explore the mechanism of this association. PMID- 12091202 TI - Psychiatric comorbidity, family dysfunction, and social impairment in referred youth with oppositional defiant disorder. AB - OBJECTIVE: The authors sought to achieve an improved understanding of the diagnosis of oppositional defiant disorder independent of its association with conduct disorder. METHOD: Family interactions, social functioning, and psychiatric comorbidity were compared in clinically referred male and female subjects with oppositional defiant disorder alone (N=643) or with comorbid conduct disorder (N=262) and a psychiatric comparison group with neither oppositional defiant disorder nor conduct disorder (N=695). RESULTS: Oppositional defiant disorder youth with or without conduct disorder were found to have significantly higher rates of comorbid psychiatric disorders and significantly greater family and social dysfunction relative to psychiatric comparison subjects. Differences between subjects with oppositional defiant disorder alone and those with comorbid conduct disorder were seen primarily in rates of mood disorders and social impairment. Oppositional defiant disorder was a significant correlate of adverse family and social outcomes when comorbid disorders (including conduct disorder) were controlled. CONCLUSIONS: These results support the validity of the oppositional defiant disorder diagnosis as a meaningful clinical entity independent of conduct disorder and highlight the extremely detrimental effects of oppositional defiant disorder on multiple domains of functioning in children and adolescents. PMID- 12091203 TI - Estrogen therapy and noncognitive psychiatric signs and symptoms in elderly patients with dementia. AB - OBJECTIVE: This study investigated the efficacy and safety of short-term estrogen therapy in decreasing noncognitive signs and symptoms of dementia in demented elderly patients. METHOD: Sixteen moderately to severely demented elderly patients with aggressive behavioral disturbances were randomly assigned to receive conjugated equine estrogens or placebo in a 4-week clinical trial. Frequency and severity of noncognitive signs and symptoms of dementia, as assessed with the Dementia Signs and Symptoms Scale, were compared between estrogen and placebo groups. Data were analyzed with intent-to-treat and regression modeling methods. RESULTS: Estrogen therapy was associated with a significantly greater improvement on the Dementia Signs and Symptoms Scale total score than placebo. All five Dementia Signs and Symptoms Scale subscale comparisons favored estrogen therapy. No adverse effects were observed. CONCLUSIONS: These preliminary data suggest that short-term estrogen therapy may safely decrease the frequency and severity of noncognitive signs and symptoms of dementia in elderly patients. PMID- 12091204 TI - Serum nerve growth factor concentration and its role in the preclinical stage of dementia. AB - OBJECTIVE: Nerve growth factor is important for the development and function of the cholinergic basal forebrain. The authors examined the hypothesis that the concentration of nerve growth factor is lower than normal in the preclinical phase of neurodegenerative dementia, especially Alzheimer's disease. METHOD: The serum nerve growth factor concentration of subjects from the Berlin Aging Study and the Berlin Memory Clinic who later developed Alzheimer's disease were compared with those of subjects who were free of dementia and subjects who were already suffering from Alzheimer's disease. RESULTS: There were 17 subjects in each group, matched for age and sex. The three groups differed in log-10 transformed mean nerve growth factor concentrations: 1.62 (SD=0.59) for the healthy comparison subjects, 0.92 (SD=0.30) for the subjects with preclinical dementia, and 1.44 (SD=0.61) for the subjects with Alzheimer's disease. CONCLUSIONS: These results support the hypothesis of disturbed nerve growth factor regulation in the serum of patients with preclinical Alzheimer's disease. Mechanisms by which these disturbances appear are unclear, but they may reflect the situation in the preclinical Alzheimer's disease brain. PMID- 12091205 TI - Antipsychotic drug treatment in the prodromal phase of schizophrenia. AB - OBJECTIVE: The safety and tolerability of short-term treatment with a low dose of risperidone was evaluated in adolescents with prodromal symptoms and a family history of schizophrenia. METHOD: Four prodromal high-risk adolescents and six first-episode patients with schizophrenia were treated with average doses of 1.0 and 1.8 mg/day of risperidone, respectively, in an 8- to 12-week open-label trial. RESULTS: No significant treatment-related adverse events were noted. Severity of thought and behavior disturbance ratings declined by about 30%; performance on a test of verbal learning improved by about 100% during treatment in both prodromal and first-episode patients, changes that achieved statistical significance despite the small group sizes. CONCLUSIONS: These findings are preliminary and should not be used to guide health care decisions at this time. Randomized controlled trials are needed to determine whether antipsychotic drug treatment of prodromal patients can delay or prevent onset or attenuate the clinical course of schizophrenia. PMID- 12091206 TI - Temporal lobe abnormalities in first-episode psychosis. AB - OBJECTIVE: The nature and time course of temporal lobe abnormalities in psychotic illness remain controversial. Confounds include disease chronicity, gender, and handedness. The present study investigated temporal substructures in right-handed male patients experiencing their first episode of psychotic illness. METHOD: Magnetic resonance imaging scans were obtained for 25 minimally treated patients experiencing their first psychotic episode and 16 healthy comparison subjects. Group differences in volumes of the hippocampus, amygdala, planum temporale, and Heschl's gyrus were tested. RESULTS: The patients had smaller bilateral hippocampal and left planum temporale volumes than the comparison subjects. Paranoid and nonparanoid patients differed in left amygdala volume. CONCLUSIONS: The authors conclude that bilateral hippocampal and left planum temporale abnormalities are present near the onset of psychosis. PMID- 12091207 TI - Depressive symptoms in adolescence as predictors of early adulthood depressive disorders and maladjustment. AB - OBJECTIVE: The authors examined the association between self-reported depressive symptoms in adolescence and mental well-being in early adulthood. METHOD: A questionnaire assessing psychosocial well-being was given to a group of subjects (N=651) in their last 3 years of high school (mean age=16.8 years) and again when these subjects reached early adulthood (mean age=21.8 years). Diagnostic interview data were obtained from a subgroup of the young adults (N=245). Adolescents' depressive symptoms were analyzed in relation to their early adulthood mental health outcome data. RESULTS: Depressive symptoms in adolescence predicted early adulthood depressive disorders (major depression and dysthymia), comorbidity, psychosocial impairment, and problem drinking. CONCLUSIONS: Depressive symptoms in adolescence deserve attention as a potential risk for early adulthood mental disorders. PMID- 12091208 TI - Elevation of the cortisol-dehydroepiandrosterone ratio in drug-free depressed patients. AB - OBJECTIVE: Elevated basal cortisol levels are a feature of depressive illness and cause deficits in learning and memory. The adrenal steroid dehydroepiandrosterone (DHEA) has antiglucocorticoid properties that may offer protection against the deleterious effects of cortisol. The authors examined the ratio of cortisol to DHEA in drug-free depressed patients and a matched comparison group. METHOD: Cortisol and DHEA were measured in saliva samples from 39 patients with unipolar depression who had been medication free for at least 6 weeks and 41 healthy comparison subjects. RESULTS: The molar cortisol-DHEA ratio was significantly higher in the depressed patients than in the healthy comparison subjects. Cortisol-DHEA ratios from saliva samples taken at 8:00 p.m. correlated positively with length of current depressive episode. CONCLUSIONS: Elevated cortisol-DHEA ratios may be a state marker of depressive illness and may contribute to the associated deficits in learning and memory. Administration of DHEA or other antiglucocorticoid treatments may reduce neurocognitive deficits in major depression. PMID- 12091209 TI - Brain-to-serum lithium ratio and age: an in vivo magnetic resonance spectroscopy study. AB - OBJECTIVE: The authors' goal was to determine if there is an association between brain-to-serum lithium ratios and age. METHOD: Lithium-7 magnetic resonance spectroscopy was used to measure in vivo brain lithium levels in nine children and adolescents (mean age=13.4 years, SD=3.6) and 18 adults (mean age=37.3, SD=9.1) with bipolar disorder. RESULTS: Serum and brain lithium concentrations were positively correlated. Younger subjects had lower brain-to-serum concentration ratios than adults: 0.58 (SD=0.24) versus 0.92 (SD=0.36). The brain to-serum concentration ratio correlated positively with age. CONCLUSIONS: These observations suggest that children and adolescents may need higher maintenance serum lithium concentrations than adults to ensure that brain lithium concentrations reach therapeutic levels. PMID- 12091210 TI - Modafinil-associated clozapine toxicity. PMID- 12091211 TI - Safety of vagus nerve stimulation with ECT. PMID- 12091213 TI - Pedophilia treated with carbamazepine and clonazepam. PMID- 12091212 TI - Galantamine for treatment-resistant schizophrenia. PMID- 12091214 TI - Narcolepsy presenting as schizophrenia. PMID- 12091215 TI - Topiramate for bulimia nervosa with bipolar II disorder. PMID- 12091216 TI - Metyrapone for Cushing's syndrome. PMID- 12091217 TI - Solar eclipse and suicide. PMID- 12091218 TI - Gabapentin-induced anorgasmia in women. PMID- 12091219 TI - Role of antidepressants in murder and suicide. PMID- 12091220 TI - The beef with atypical antipsychotics. PMID- 12091221 TI - Pregnancy in women with eating disorders. PMID- 12091222 TI - Change in criterion for paraphilias in DSM-IV-TR. PMID- 12091239 TI - Nomination of a candidate susceptibility gene in sarcoidosis: the complement receptor 1 gene. PMID- 12091240 TI - Heme oxygenase-1: the "emerging molecule" has arrived. AB - Organisms on our planet have evolved in an oxidizing environment that is intrinsically inimical to life, and cells have been forced to devise means of protecting themselves. One of the defenses used most widely in nature is the enzyme heme oxygenase-1 (HO-1). This enzyme performs the seemingly lackluster function of catabolizing heme to generate bilirubin, carbon monoxide, and free iron. Remarkably, however, the activity of this enzyme results in profound changes in cells' abilities to protect themselves against oxidative injury. HO-1 has been shown to have anti-inflammatory, antiapoptotic, and antiproliferative effects, and it is now known to have salutary effects in diseases as diverse as atherosclerosis and sepsis. The mechanism by which HO-1 confers its protective effect is as yet poorly understood, but this area of invetsigation is active and rapidly evolving. This review highlights current information on the function of HO-1 and its relevance to specific pulmonary and cardiovascular diseases. PMID- 12091241 TI - Complement receptor 1 gene polymorphisms in sarcoidosis. AB - Sarcoidosis is likely to result from exposure of genetically susceptible hosts to environmental agents. Erythrocyte (E) complement receptor 1 (CR1) is a membrane protein mediating the transport of immune complexes (ICs) to phagocytes, and at least three polymorphisms on the CR1 gene are related to erythrocyte surface density of CR1 molecules, in turn related to the rate of IC clearance from circulation. We hypothesized that sarcoidosis could be associated with increased frequency of the CR1 gene alleles coding for reduced CR1/E ratio. We studied 91 sarcoid patients and two control groups: 94 healthy volunteers and 71 patients with chronic obstructive pulmonary disease (COPD). Three polymorphic sites of CR1 gene, His1208Arg, intron 27 HindIII/RFLP, and Pro1827Arg, were analyzed. The three polymorphisms were in linkage disequilibrium. The GG genotype for the Pro1827Arg (C(5507)G) polymorphism was significantly associated with sarcoidosis in comparison to both control groups (odds ratio [OR] = 3.13; 95% confidence interval [CI] 1.49-6.69 versus healthy control subjects, and OR= 2.82, 95% CI 1.27-6.39 versus COPD control subjects). The same genotype was particularly associated to disease in females (OR = 7.05; 95% CI 3.10-16.61 versus healthy control subjects). These findings agree with speculations on the role of CR1 gene as a possible susceptibility factor. PMID- 12091242 TI - Lymphocyte activation in the lungs of SP-D null mice. AB - Surfactant protein D (SP-D) appears to play an important role in regulating local pulmonary inflammatory responses to pathogens. There is also in vitro evidence that SP-D may suppress local T cell responses. However, the role of SP-D in regulating T cell responses directly in the lung has not been previously evaluated in vivo. SP-D(-)(/-) mice demonstrate peribronchial and perivascular accumulations of lymphocytes. Therefore, we investigated the functional status and abundance of intrapulmonary lymphocytes in SP-D(-)(/-) mice. By morphometric analysis, SP-D(-)(/-) mice demonstrated increased numbers of airway- and vessel associated lymphocytes without increases in interstitial lymphocytes. There was increased proliferative activity of lymphocytes isolated by enzymatic disassociation of minced lung. Flow cytometry was used to determine the number and functional activation status of intrapulmonary CD4(+) and CD8(+) T cells, as well as B cells and NK cells. Cytokine expression patterns in lung tissues were evaluated using RNase protection assays, reverse transcriptase/polymerase chain reaction, and enzyme-linked immunosorbent assay. There was marked T cell activation in the lungs of SP-D(-)(/-) mice, as reflected by an increased percentage of both CD4(+) and CD8(+) T cells expressing CD69 and CD25. BAL CD4 lymphocytes were increased and the fraction expressing CD69 was also increased. Although there were increases in BAL CD8 lymphocytes, apparent increases in CD69 positive CD8 lymphocytes did not reach statistical significance. In contrast, splenic T cells were not activated in SPD(-)(/-) mice. Of the proinflammatory cytokines evaluated, only interleukin (IL)-12 and IL-6 expression were consistently upregulated in the lungs of SPD(-)(/-) mice. Increased IL-2 expression was apparent but did not reach statistical significance. We conclude that the lack of local pulmonary production of SP-D leads to a state of persistent T cell activation, possibly in response to exogenous antigens. This study therefore provides further evidence of the important local immunoregulatory role of SP-D in vivo. PMID- 12091243 TI - Interaction of alveolar macrophages and airway epithelial cells following exposure to particulate matter produces mediators that stimulate the bone marrow. AB - Exposure to ambient air pollution particles with a diameter of < 10 microm (PM(10)) has been associated with increased cardiopulmonary morbidity and mortality. We postulate that these adverse health effects are related to proinflammatory mediators produced in the lung and released into the circulation where they initiate a systemic inflammatory response. The present study was designed to determine if alveolar macrophages (AMs) and primary human bronchial epithelial cells (HBECs) interact to amplify the production of certain cytokines when exposed to ambient PM(10) (EHC-93). Candidate cytokines were measured at the mRNA level using a RNase protection assay and at the protein level by enzyme linked immunosorbent assay (ELISA). When AM/HBEC cocultures were exposed to 100 microg/ml of PM(10), levels of tumor necrosis factor (TNF)-alpha, granulocyte macrophage colony stimulating factor (GM-CSF), interleukin (IL)-1beta, IL-6, leukemia inhibitory factor (LIF), oncostatin M (OSM), and IL-8 mRNA increased within 2 h (P < 0.05) and 8 h following exposure compared with control cells. GM CSF mRNA expression was more rapidly induced in cocultured cells compared with HBECs or AMs alone. The concentrations of TNF-alpha, GM-CSF, IL-1beta, IL-6, and IL-8 in the cocultured supernatants collected after 24 h PM(10) exposure increased significantly compared with control cells. There was a significant synergistic effect between AMs and HBECs in the production of GM-CSF and of IL-6 (P < 0.05). Instillation of supernatants from HBECs cultured with PM(10) into lungs of rabbits failed to increase circulating band cell counts or stimulate the bone marrow. However, those from AM/HBEC cocultures exposed to PM(10) increased circulating band cell counts (P < 0.05) and shortened the transit time of polymorphonuclear leukocytes (PMNs) through the bone marrow compared with control co-cultures (P < 0.01). These results suggest that the interaction between AMs and HBECs during PM(10) exposure contributes to the production of mediators that induce a systemic inflammatory response. PMID- 12091244 TI - Altered guanylyl-cyclase activity in vitro of pulmonary arteries from fetal lambs with congenital diaphragmatic hernia. AB - Nitric oxide (NO) plays a major role in the modulation of perinatal pulmonary vascular tone. Congenital diaphragmatic hernia (CDH), a major cause of severe persistent pulmonary hypertension of the newborn (PPHN), is often refractory to inhaled NO. Alterations in NO/cyclic guanosine 3',5' monophosphate (cGMP) mediated pulmonary vasodilatation may contribute to PPHN in CDH. We assessed NO/cGMP-mediated pulmonary vasorelaxation in vitro in 140-d gestational lamb fetuses with surgically created left CDH (term = 147 d) to age-matched controls. Relaxation of fourth generation intralobar pulmonary artery rings in response to the endothelium-dependent vasodilator, acetylcholine (ACh), and to the specific inhibitor of cGMP-phosphodiesterase (PDE), zaprinast, did not differ between the two groups. By contrast, relaxation in response to the calcium ionophore A23187 was impaired in CDH as compared with control animals. Relaxation in response to the NO donor sodium nitroprusside (SNP) (a direct activator of soluble guanylyl cyclase [sGC]) was also impaired in CDH animals as compared with controls. Repeating the challenge increased vasorelaxation in response to SNP in CDH as compared with control animals. Immunohistochemistry revealed the presence of endothelial NO-synthase in the endothelium of pulmonary arteries from both control and CDH animals. We conclude that endothelium-dependent vasodilatation in response to ACh and A23187 was differently affected in the fetal surgical CDH lamb model. Furthermore, activity of sGC but not that of PDE was impaired in CDH animals. PPHN and decreased inhaled NO responsiveness in CDH may involve decreased sGC activity. PMID- 12091245 TI - Identification of a novel IL-6 isoform binding to the endogenous IL-6 receptor. AB - Interleukin (IL)-6 is a multifunctional cytokine showing a wide variety of biologic functions on various tissues. Extracellular IL-6 signals through heterohexameric complex formation with IL-6 receptor-alpha (IL-6Ralpha) and IL-6 receptor-beta (IL-6Rbeta). In analogy to cytokines IL-2 and IL-4, we investigated the expression of IL-6 splice variants in lung tissue and cultivated fibroblasts. In human lung specimens, four different IL-6 transcripts were characterized as follows: native IL-6; IL-6 missing either exon 2 (IL-6Delta2), exon 4 (IL 6Delta4), or missing both; and exons 2 and 4 (IL-6Delta2,4). Only native IL-6 and IL-6Delta4 encoded for proteins of ~ 26 and 17 kD, respectively. Although the overall structure and most functional sites of the IL-6Delta4 protein were predicted to be maintained, IL-6Delta4 was found to lack two amino acids necessary for IL-6/IL-6 homodimerization as well as two of the six amino acids required for interaction with IL-6Rbeta. Receptor mobility shift assays confirmed that the new isoform formed a stable complex with IL-6Ralpha; however, no interaction with IL-6Rbeta was observed. Thus, IL-6Delta4 is likely to compete with native IL-6 for IL-6Ralpha binding but fails to transmit IL-6Rbeta-mediated signaling. PMID- 12091246 TI - HDL-holoparticle uptake by alveolar type II cells: effect of vitamin E status. AB - Alveolar type II cells accumulate vitamin E preferentially from high-density lipoproteins (HDL) and express at least three receptors that are specific for HDL. The expression of these receptors increases in response to vitamin E deficiency. Beside receptors for specific lipid transfer from HDL, cubilin and megalin, several other receptors that mediate HDL-particle uptake were found in the lung. We hypothesize that alveolar type II cells also exhibit the HDL particle uptake and that this process can be regulated by the vitamin E status. By confocal laser microscopy and flow cytometry we showed that type II cells accumulate protein-labeled HDL-particle. Vitamin E depletion in rats increased HDL-particle uptake in alveolar type II cells and the expression of megalin. The expression of cubilin did not change. Refeeding with vitamin E reversed HDL particle uptake and megalin expression. Long-time incubation of type II cells with phorbol myristyl acetate (PMA) reduced HDL-holoparticle uptake and megalin expression. We assume that alveolar type II cells exhibit HDL-holoparticle uptake mediated by megalin and cubilin. Megalin represents the regulated element of the megalin/cubilin receptor-cooperation and can be modulated by protein kinase C. PMID- 12091248 TI - Identification of leptin receptors in lung and isolated fetal type II cells. AB - Leptin is a cytokine involved in regulation of the satiety response. Receptors for this protein have been identified in brain as well as many other peripheral tissues. Some of the highest levels of receptor concentration occur in the lung. Considering the cellular diversity of lung, neither the localization nor the function of leptin in pulmonary tissues has been delineated. The purpose of the present study was to determine if fetal and adult rabbit lung displayed specific binding for leptin, to identify the binding sites, and to explore a potential functional role for leptin in lung surfactant production. Frozen sections of adult and fetal rabbit (24th gestational day) lung were prepared and incubated with increasing concentrations of [125I]leptin in the presence or absence of 1 microM-unlabeled leptin. Sections were removed and radioactivity measured. Concurrently, sections were coated with nuclear Trac emulsion and incubated in the dark at -30 degrees C. Lung showed specific binding for leptin. Microscopically, [125I]leptin was localized to acinar-lining epithelium of developing fetal lung. Larger cells within the epithelial layer appeared to bind leptin more avidly than adjacent cells. Antibodies to the leptin receptor were used to identify binding sites in adult lung and isolated fetal lung type II cells. In adult lung, both the K20 (against the extracellular amino-terminal) and the M18 antibody (against the intracellular carboxy-terminal) displayed several binding sites. In contrast, the isolated fetal type II cells showed only a single binding site for both antibodies. The apparent molecular mass of the receptor using the K20 antibody appeared to be approximately 125 kD. A protein of similar mass bound the M18 antibody suggesting that functional receptor is present in lung and expressed by fetal type II cells. Incubation of isolated fetal type II cells with leptin (0.01-10 microg/ml) stimulated [3H]choline incorporation in disaturated phosphatidylcholine. These results show that fetal and adult lung bind leptin specifically, and fetal type II cells in particular, may be responsive to leptin stimulation of phospholipid production. Leptin may therefore be important in regulating maturation of cells of the fetal lung. PMID- 12091247 TI - BMK1 (ERK5) regulates squamous differentiation marker SPRR1B transcription in Clara-like H441 cells. AB - Various toxicants and carcinogens upregulate the expression of small proline-rich protein 1B (SPRR1B), a squamous differentiation marker, in bronchial epithelial cells both in vivo and in vitro. We have recently shown that phorbol 13-myristate 12-acetate (PMA)-stimulated SPRR1B transcription in Clara-like H441 cells is mainly mediated by activator protein-1 (AP-1) and c-Jun N-terminal kinase-1 (JNK1). Though mitogen-activated protein kinase (MAPK) kinase (MEK)-1/2 pathway inhibitors strongly suppressed both basal and PMA-inducible SPRR1B transcription, overexpression of dominant negative (dn) forms of extracellular signal-regulated kinase (ERK)-1 and/or -2 did not have any significant effect indicating the involvement of another ERK-like MAPK in this pathway. Here, we report for the first time the involvement of ERK5 in PMA-inducible SPRR1B transcription in H441 cells. PMA significantly induced ERK5 activation in H441 cells. Overexpression of dn-ERK5 strongly suppressed both basal and PMA-inducible SPRR1B transcription, whereas wild-type ERK5 upregulated it. Consistent with this, a mutant form of MEK 5, an upstream activator of ERK5, strongly suppressed PMA-inducible promoter activity. However, coexpression of c-Jun restored promoter activation suppressed by dn-ERK5. Thus, in addition to JNK1, the activation of MEK5-ERK5 MAPK pathway probably plays a pivotal role in transcriptional regulation of AP-1-mediated SPRR1B expression in the distal bronchiolar region. PMID- 12091249 TI - Apoptosis underlies immunopathogenic mechanisms in acute silicosis. AB - We investigated immunopathogenic roles for apoptosis in acute murine silicosis. Intratracheal silica instillation induced pulmonary inflammation and enlarged thoracic lymph nodes. Lymphocytes from silica-exposed lymph nodes showed reduced mitogenic responses to T cell receptor (TCR) stimulation, and markedly increased activation-induced cell death, compared with control lymphocytes from saline exposed lymph nodes. CD4(+) T cell death was mediated by Fas ligand, because CD4(+) T cells from Fas ligand-deficient gld mice did not undergo activation induced apoptosis. Silica deposition also resulted in increased apoptosis associated with inflammatory infiltrates in lung parenchyma. In vivo treatment with caspase inhibitors reduced neutrophil accumulation, and alleviated inflammation in the lungs of silica-treated mice. These results suggest that silica-induced apoptosis plays an inflammatory role in the lung parenchyma, and creates immunologic abnormalities in regional lymph nodes, with pathogenic implications for the host. PMID- 12091250 TI - Autocrine ligands for the epidermal growth factor receptor mediate interleukin-8 release from bronchial epithelial cells in response to cigarette smoke. AB - Airway neutrophilia is a prominent feature of chronic obstructive pulmonary disease. As cigarette smoke (CS) and epidermal growth factor (EGF) both cause release of interleukin-8 (IL-8) from epithelial cells in vitro, we investigated whether autocrine ligands for the EGF receptor (EGFR) are involved in this proinflammatory response to CS. NCI-H292 or primary bronchial epithelial cells were cultured with or without cigarette smoke extract (CSE) or EGF for 6-48 h. We then tested culture supernatants for lactate dehydrogenase activity to assess cell viability, and for IL-8 and EGFR ligands by ELISA; quantitative RT-PCR was used to measure IL-8 and EGFR ligand mRNA. EGF and low concentrations of CSE both promoted cell survival and caused enhanced transcription and release of IL-8. Similarly, levels of mRNA encoding transforming growth factor alpha (TGF-alpha), heparin-binding EGF-like growth factor, and amphiregulin (AR) were increased, as was shedding of TGF-alpha and AR protein into the culture medium. With the exception of AR gene transcription, the CS-induced responses were blocked by the EGFR-selective kinase inhibitor AG1478. Furthermore, ~ 45% of CS-induced IL-8 release was inhibited by a neutralising anti-EGFR. Our data indicate that secretion of IL-8 in response to CSE is dependent on EGFR activation and that autocrine production of TGF-alpha makes a substantial contribution to this response. PMID- 12091251 TI - Silica-induced apoptosis in murine macrophage: involvement of tumor necrosis factor-alpha and nuclear factor-kappaB activation. AB - Alveolar macrophages play a critical role in silica-induced lung fibrosis. Silica exposure induces tumor necrosis factor (TNF)-alpha release and nuclear factor (NF)-kappaB activation, and apoptotic mechanisms have been implicated in silica induced pathogenesis. To characterize potential relationships between these signaling events, we studied their induction in two murine macrophage cell lines. The RAW 264.7 macrophage cell line was more sensitive, and the IC-21 macrophage cell line more tolerant to silica exposure (0.2 or 1 mg/ml for 6 h) as evidenced by significantly higher apoptotic responses in RAW 264.7 (P < 0.05). RAW 264.7 macrophages exhibited enhanced TNF-alpha production and NF-kappaB activation in response to silica, whereas IC-21 macrophages did not produce TNF-alpha in response to silica and did not induce NF-kappaB nuclear binding. Inhibition of NF kappaB in RAW 264.7 cells with BAY11-7082 significantly increased apoptosis while inhibiting TNF-alpha release. In addition, TNF-alpha and NF-kappaB activation, but not apoptosis, were induced by lipopolysaccharide (LPS) in both cell lines, and NF-kappaB inhibition reduced LPS-induced TNF-alpha release. These data suggest that TNF-alpha induction is dependent on NF-kappaB activation in both cell lines. However, silica can induce apoptosis in murine macrophages, independently of TNF-alpha stimulation, as in IC-21 macrophages. Furthermore, NF kappaB activation in macrophages may play dual roles, both pro- and antiapoptotic during silica injury. PMID- 12091252 TI - Inhibition of glutamine synthetase in a549 cells during hyperoxia. AB - High oxygen concentrations are used in the treatment of acute respiratory distress syndrome and hyaline membrane disease. Hyperoxia, however, can damage alveolar epithelial cells through the release of free oxygen radicals. Supplemental glutamine (Gln) has recently been shown to increase survival of A549 cells, a distal epithelial cell line, during hyperoxia (). We found that supplemental Gln (Gln+) is essential for cell growth in A549 cells. In room air, cells without supplemental Gln (Gln-) survived with BCL-2 levels similar to those of Gln+ cells, but cell growth was minimal. We also evaluated the role of glutamine synthetase (GS) in A549 cells during hyperoxia. L-methionine sulfoximine (MSO), an irreversible inhibitor of GS, was added to Gln+ and Gln- cells. In hyperoxia, Gln- cells had greater survival then Gln- cells treated with MSO. Supplemental Gln could rescue cells in hyperoxia from the effect of MSO, suggesting that GS, through the endogenous synthesis of Gln, could attenuate hyperoxic cell injury. In hyperoxia, cells treated with 10-mM concentrations of Gln had increased survival compared with cells receiving 2-mM concentrations. The higher concentration of Gln, however, did not decrease the percentage of cells undergoing necrosis. PMID- 12091253 TI - Role of MAP kinase activation in interleukin-8 production by human BEAS-2B bronchial epithelial cells submitted to cyclic stretch. AB - Overstretching the airways during positive pressure mechanical ventilation or attacks of acute severe asthma is associated with important biologic responses. Interleukin (IL)-8-dependent neutrophil recruitment seems to play a critical role in the process of mechanical stress-induced airway inflammation. Herein, we show that human bronchial epithelial BEAS-2B cells submitted to cyclic stretch in vitro produce IL-8, at both the mRNA and protein levels. This cellular stress "turns on" activator protein (AP)-1 and cyclic AMP (cAMP)-responding elements. The mitogen-activated protein (MAP) kinases (MAPK) p44/42, SAPK/JNK, and p38 were all rapidly activated (phosphorylated) after the initiation of the cyclic strain (5-10 min). The blockade of p38 with the pharmacologic inhibitor SB203580 abrogated IL-8 production by cell stretching, and an inhibitor of the p44/42 pathway, PD98059, partially inhibited the IL-8 response. A nonspecific tyrosine kinase inhibitor, genistein, also blocked the stretch-induced IL-8 production. This suggests that MAPK, and p38 in particular, are proximal and key intracellular signaling molecules mediating cell activation in response to cyclic stretch, a mechanical strain similar to that applied to lung epithelial cells during mechanical ventilation. Pharmacologic inhibition of the p38 pathway holds promise as a new therapeutic avenue in ventilated patients. PMID- 12091254 TI - Outcome measurement in mental health--welcome to the revolution. PMID- 12091255 TI - Noble experiment II: selective prohibition. PMID- 12091256 TI - Explanatory models for mental distress: implications for clinical practice and research. PMID- 12091257 TI - Outcomes research in mental health. Systematic review. AB - BACKGROUND: Outcomes research involves the secondary analysis of data collected routinely by clinical services, in order to judge the effectiveness of interventions and policy initiatives. It permits the study of large databases of patients who are representative of 'real world' practice. However, there are potential problems with this observational design. AIMS: To establish the strengths and limitations of outcomes research when applied in mental health. METHOD: A systematic review was made of the application of outcomes research in mental health services research. RESULTS: Nine examples of outcomes research in mental health services were found. Those that used insurance claims data have information on large numbers of patients but use surrogate outcomes that are of questionable value to clinicians and patients. Problems arise when attempting to adjust for important confounding variables using routinely collected claims data, making results difficult to interpret. CONCLUSIONS: Outcomes research is unlikely to be a quick or cheap means of establishing evidence for the effectiveness of mental health practice and policy. PMID- 12091258 TI - Assessing the evidence on case management. AB - BACKGROUND: Evidence on the impact of case management is contradictory. AIMS: To discuss two different systematic reviews (one conducted by the authors and one conducted through the Cochrane collaboration) that came to contradictory conclusions about the impact of case management in mental health services. METHOD: We summarised the findings of the two reviews with respect to case management effectiveness, examined key methodological differences between the two approaches and discuss the impact of these on the validity of the results. RESULTS: The differences in conclusions between the two reviews result from the differences in inclusion criteria, namely non-randomised trials, data from unpublished scales and data from variables with skewed distributions. The theoretical and empirical effects of these are discussed. CONCLUSIONS: Systematic reviewers may face a trade-off between the application of strict criteria for the inclusion of studies and the amount of data available for analysis and hence statistical power. The available research suggests that case management is generally effective. PMID- 12091259 TI - Influence of supplementary vitamins, minerals and essential fatty acids on the antisocial behaviour of young adult prisoners. Randomised, placebo-controlled trial. AB - BACKGROUND: There is evidence that offenders consume diets lacking in essential nutrients and this could adversely affect their behaviour. AIMS: To test empirically if physiologically adequate intakes of vitamins, minerals and essential fatty acids cause a reduction in antisocial behaviour. METHOD: Experimental, double-blind, placebo-controlled, randomised trial of nutritional supplements on 231 young adult prisoners, comparing disciplinary offences before and during supplementation. RESULTS: Compared with placebos, those receiving the active capsules committed an average of 26.3% (95% CI 8.3-44.33%) fewer offences (P=0.03, two-tailed). Compared to baseline, the effect on those taking active supplements for a minimum of 2 weeks (n=172) was an average 35.1% (95% CI 16.3 53.9%) reduction of offences (P<0.001, two-tailed), whereas placebos remained within standard error. CONCLUSIONS: Antisocial behaviour in prisons, including violence, are reduced by vitamins, minerals and essential fatty acids with similar implications for those eating poor diets in the community. PMID- 12091260 TI - Efficacy of citalopram in the prevention of recurrent depression in elderly patients: placebo-controlled study of maintenance therapy. AB - BACKGROUND: The highly recurrent nature of major depression in the young and the elderly warrants long-term antidepressant treatment. AIMS: To compare the prophylactic efficacy of citalopram and placebo in elderly patients; to evaluate long-term tolerability of citalopram. METHOD: Out-patients, > or =65 years, with unipolar major depression (DSM-IV: 296.2 x or 296.3 x) and Montgomery-Asberg Depression Rating Scale score > or =22 were treated with citalopram 20-40 mg for 8 weeks. Responders continued on their final fixed dose of citalopram for 16 weeks before randomisation to double-blind treatment with citalopram or placebo for at least 48 weeks. RESULTS: Nineteen of the 60 patients using citalopram v. 41 of the 61 patients using placebo had recurrence. Time to recurrence was significantly different between citalopram- and placebo-patients, in favour of citalopram (log-rank test, P<0.0001). Long-term treatment was well tolerated. CONCLUSIONS: Long-term treatment with citalopram is effective in preventing recurrence of depression in the elderly and is well tolerated. PMID- 12091261 TI - Estimating the relationship between disease progression and cost of care in dementia. AB - BACKGROUND: Previous studies have shown a positive relationship between disease severity and cost. AIMS: To explore the factors affecting time to institutionalisation and estimate the relationship between the costs of care and disease progression. METHOD: Retrospective analysis of a longitudinal data-set for a cohort of 100 patients diagnosed with Alzheimer's disease or vascular dementia. RESULTS: Changes in both Mini-Mental State Examination (MMSE) and Barthel scores have independent and significant marginal effects on costs. Each one-point decline in the MMSE score is associated with a pound sterling 56 increase in the four-monthly costs, whereas each one-point fall in the Barthel index is associated with a pound sterling 586 increase in costs. CONCLUSIONS: It may be inappropriate for economic models of disease progression in dementia to be based solely on measures of cognitive change. MMSE and the Barthel index are independent significant predictors of time to institutionalisation and cost of care, but changes in the Barthel index are particularly important in predicting costs outside institutional care. PMID- 12091262 TI - Pathways to care in children at risk of attention-deficit hyperactivity disorder. AB - BACKGROUND: There is underdiagnosis of and low use of specialist services for attention-deficit hyperactivity disorder (ADHD). AIMS: To quantify the filters in the help-seeking pathway through primary care and to investigate factors influencing progress for children at risk of ADHD. METHOD: A total of 127 children (5-11 years old) with pervasive hyperactivity who passed each filter (primary care attendance and general practitioner (GP) recognition of disorder) were compared with those who had not. RESULTS: Primary care attendance was only associated with parental perception of the behaviour as problematic (OR 2.11; 95% CI 1.11-4.03). However, GP recognition was related to both parent and child factors - parental request for referral (OR 20.83; 95% CI 3.05-142.08) and conduct problems (OR 1.48; 95% CI 1.04-2.12). GP non-recognition was the main barrier in the pathway to care; following recognition, most children were referred. CONCLUSIONS: Parents can be regarded as the main gatekeepers for access to specialist services. PMID- 12091263 TI - Sexual dysfunction in patients taking conventional antipsychotic medication. AB - BACKGROUND: Antipsychotic drugs are associated with sexual dysfunction but the mechanisms are poorly understood. AIMS: To ascertain the frequency of sexual dysfunction in patients taking conventional antipsychotics and to determine the possible underlying mechanisms. METHOD: Sexual dysfunction was assessed in 101 patients receiving conventional antipsychotic medication, 57 normal controls and 55 controls attending a sexual dysfunction clinic. RESULTS: Sexual dysfunction occurred in 45% of patients taking antipsychotic medication, 17% of normal controls and 61% of controls attending a sexual dysfunction clinic. Sexual dysfunction was associated with autonomic side-effects in normoprolactinaemic males, but the presence of hyperprolactinaemia overrode other causes of sexual dysfunction. For women, hyperprolactinaemia was the main cause of sexual dysfunction. CONCLUSIONS: Conventional anti-psychotic medications cause significant levels of sexual dysfunction. Clinicians should routinely enquire about sexual symptoms prior to the prescription of antipsychotics and on follow up. PMID- 12091264 TI - Neurasthenia: prevalence, disability and health care characteristics in the Australian community. AB - BACKGROUND: Neurasthenia imposes a high burden on primary medical health care systems in all societies. AIMS: To determine the prevalence of ICD-10 neurasthenia and associated comorbidity, disability and health care utilisation. METHOD: Utilisation of a national sample of Australian households previously surveyed using the Composite International Diagnostic Interview and other measures. RESULTS: Prolonged and excessive fatigue was reported by 1465 people (13.29% of the sample). Of these, one in nine people meet current ICD-10 criteria for neurasthenia. Comorbidity was associated with affective, anxiety and physical disorders. People with neurasthenia alone (<0.5% of the population) were less disabled and used less services than those with comorbid disorders. CONCLUSIONS: Fatigue is frequent in the Australian community and is common in people attending general practice. Neurasthenia is disabling and demanding of services largely because of its comorbidity with other mental and physical disorders. Until a remedy for persistent fatigue is provided, doctors should take an active psychological approach to treatment. PMID- 12091265 TI - Economic impact of personality disorders in UK primary care attenders. AB - BACKGROUND: The economic impact of personality disorders on UK health services is unknown. AIMS: To test the hypothesis that people with personality disorders have higher mean health and non-health costs compared with those without personality disorders. METHOD: Prospective cohort study design. A total of 303 general practice attenders were followed-up I year after they had been assessed for the presence of personality disorders. Costs were estimated in pound sterling at 1999 price levels. RESULTS: The mean total cost for patients with personality disorders was pound sterling 3094 (s.d.=5324) compared with pound sterling 1633 (s.d.=3779) for those without personality disorders. Personality disorders were not independently associated with increased costs. Multivariate analyses identified the presence of a significant interaction between personality disorders and common mental disorders and increased total costs (coefficient=499, 95% CI 180.1-626.2, P=0.002). CONCLUSIONS: Personality disorders are not independently associated with increased costs. An interaction between personality disorders and common mental disorders significantly predicts increased total costs. PMID- 12091266 TI - Behavioural symptoms among people with severe and profound intellectual disabilities: a 26-year follow-up study. AB - BACKGROUND: Very little is known about the natural history of challenging behaviour and psychiatric disorder in people with severe and profound degrees of intellectual disability. AIMS: To clarify the natural history of challenging behaviour and psychiatric disorder in this population through a longterm prospective cohort study over a 26-year period. METHOD: One hundred individuals with severe or profound intellectual disability were randomly selected in 1975. Their behaviour was recorded through carer and psychiatrist ratings using the Modified Manifest Abnormality Scale of the Clinical Interview Schedule. The presence and severity of psychiatric disorder were also recorded. The study was repeated in 1981/82 and 1992/93. We repeated the study again in 2001, supplementing the original observational data with the Checklist of Challenging Behaviour. RESULTS: Behavioural symptomatology is remarkably persistent, particularly stereotypy, emotional abnormalities, eye avoidance and overactivity, although the severity of overall psychiatric disorder does show some abatement through time. CONCLUSIONS: These findings influence the prospects of success in relocating adults with severe and profound degrees of intellectual disability back into the community. PMID- 12091267 TI - Sudden death and suicide: a comparison of brain weight. AB - BACKGROUND: Recent evidence suggests that the brain weight of individuals over the age of 60 who commit suicide is significantly higher than in those who die of natural causes. AIMS: To ascertain whether brain weight is different in people of a younger age who commit suicide than in those who die accidentally. METHOD: A retrospective review of post-mortem reports collecting height, weight and brain weight in 100 suicide victims (87 males, mean age 38.5 years) and 100 age/gender matched controls who died accidentally or of natural causes (87 males, mean age 38.7 years). Comparison by t-test was made of brain weight in isolation as well as brain weight corrected for height, weight and body mass index. RESULTS: These results reveal no significant difference in brain weight in suicide cases compared to the general population (P > 0.05). The brain weight of those who died by hanging was significantly higher than of those who died by overdose. CONCLUSIONS: Whatever the significant neuropsychiatric elements are that influence suicidal behaviour, they do not consistently affect brain weight in the population studied. PMID- 12091268 TI - Personality disorder. PMID- 12091269 TI - Personality disorder. PMID- 12091270 TI - Personality disorder. PMID- 12091271 TI - Patient adherence with antidepressant treatment. PMID- 12091272 TI - Patient adherence with antidepressant treatment. PMID- 12091273 TI - Recruitment of psychiatrists. PMID- 12091274 TI - Cost of somatisation. PMID- 12091278 TI - Management issues in CLD of prematurity. AB - CLD is one of the major problems in neonatal medicine. Clinicians struggle to provide optimal care because of limited evidence. Many questions still need to be answered; meanwhile many aspects of management will remain best practice rather than evidence based. PMID- 12091279 TI - Update on modalities of mechanical ventilators. AB - Recent advances in ventilator technology have often not been confirmed by randomised trials and instead serious shortcomings have been highlighted. Ventilation modes should only be introduced into routine clinical practice when proved efficacious in appropriately designed studies and no adverse outcomes identified by long term follow up. PMID- 12091280 TI - Role of lung function testing in the management of mechanically ventilated infants. AB - The mechanical characteristics of the ventilated lung can only be interpreted when the volume of the lung, the elastic properties, and the degree of airway obstruction have been accurately quantified by pulmonary function testing. More gentle ventilation strategies (permissive hypercapnia) are used, and the efficacy of mechanical ventilation can be verified in the intensive care unit. Pulmonary function testing brings new insights, awareness, and applications, but its limitations need to be taken into account when interpreting the acquired data. PMID- 12091281 TI - Oxygen therapy for infants with chronic lung disease. AB - Supplemental oxygen is a safe and effective treatment for infants with established chronic lung disease who are not at risk of further progression of retinopathy of prematurity (ROP). Oxygen saturations of < 92% should be avoided and a target range of at least 94-96% aimed for. The saturation target range for very preterm infants at risk of developing ROP is more controversial, but the therapeutic index is probably considerably narrower. PMID- 12091282 TI - Monitoring cardiovascular function in infants with chronic lung disease of prematurity. AB - In addition to persistent airways disease, survivors of premature birth with chronic lung disease are at risk of cardiovascular sequelae, including pulmonary hypertension, systemic hypertension, left ventricular hypertrophy, and exercise intolerance. The major treatment of pulmonary hypertension is supplemental oxygen, but drugs such as calcium channel blockers may also be required. The use of inhaled nitric oxide for its long term management is being investigated PMID- 12091283 TI - Prenatal diagnosis of structural heart disease: does it make a difference to survival? PMID- 12091284 TI - Diagnosis, prevention, and management of catheter related bloodstream infection during long term parenteral nutrition. PMID- 12091285 TI - Blood cultures in newborns and children: optimising an everyday test. AB - Effective use of blood cultures is a key component of the management of septic newborns and children. The technical and practical aspects of paediatric practice and the heightened susceptibility of children to infection because of immunological immaturity make automatic extrapolation of adult data difficult and potentially unfounded. PMID- 12091286 TI - Neonatal shaken baby syndrome: an aetiological view from Down Under. PMID- 12091287 TI - Poor outcome of very low birthweight babies with serious congenital heart disease. AB - OBJECTIVE: To evaluate incidence and mortality of congenital heart disease in very low birthweight babies. METHOD: Retrospective analysis of a 12 year period. RESULTS: Forty seven babies were diagnosed with severe congenital heart disease. The most common lesions were ventricular septal defect and coarctation of the aorta. Mortality attributed to congenital heart disease was 32%. Coarctation of the aorta, the second most common lesion, was fatal in 62% of cases. Closure of a patent ductus arteriosus with indomethacin proved to be detrimental in babies with undiagnosed coarctation, causing rapid deterioration in some. CONCLUSION: Very low birthweight neonates with severe congenital heart disease have a higher mortality than babies with higher birth weight. A contributing factor is closure of a patent ductus arteriosus if an underlying lesion has not been recognised. This could be of significance if the use of prophylactic treatment with indomethacin becomes more common. PMID- 12091288 TI - Procollagen I C-propeptide in the cerebrospinal fluid of neonates with posthaemorrhagic hydrocephalus. AB - BACKGROUND: The pathogenesis of posthaemorrhagic hydrocephalus (PHHC) following intraventricular haemorrhage (IVH) in premature infants includes a fibroproliferative reaction leading to arachnoidal fibrosis, ultimately causing malresorption of cerebrospinal fluid (CSF) at the arachnoid villi. AIMS: To determine whether an increased concentration of the carboxyterminal propeptide of type I procollagen (PICP) in the CSF of neonates after IVH reflects the activation of collagen synthesis preceding the manifestation of PHHC. METHODS: From 20 neonates with PHHC (median birth weight 740 g, median gestational age 25+1 weeks), 52 CSF samples were collected. CSF samples of four neonates (median birth weight 2170 g, median gestational age 32+4 weeks) with congenital non haemorrhagic hydrocephalus served as controls. PICP was measured by radioimmunoassay. RESULTS: PICP in CSF taken at the start of external CSF drainage (median day 21, range 17-25 days postnatal age) was significantly increased (median 851.5, range 153.5-1944 microg/l) compared with controls (median 136.1, range 33.8-169.5 microg/l). CSF concentrations of PICP declined until permanent shunt placement (median day 70, range days 41-113). CONCLUSION: In neonates who develop PHHC, significant elevation of PICP concentration in the CSF is present 3-4 weeks after IVH. It reflects the increase of local type I collagen turnover, thereby correlating with manifestation of PHHC. PMID- 12091289 TI - Posthaemorrhagic ventricular dilatation in the premature infant: natural history and predictors of outcome. AB - OBJECTIVE: To investigate the natural history and predictors of outcome of posthaemorrhagic ventriculomegaly in the very low birthweight (VLBW) infant. METHODS: All VLBW infants admitted between September 1994 and September 1997 to the neonatal intensive care units of Brigham and Women's Hospital (Boston), Children's Hospital (Boston), and Christchurch Women's Hospital (New Zealand) with germinal matrix intraventricular haemorrhage (IVH) were identified. All charts and ultrasound scans were reviewed to define the natural history and perinatal and/or postnatal factors of value in prediction of the course of posthaemorrhagic ventriculomegaly. Progressive ventricular dilatation (PVD) was defined from the results of serial cranial ultrasound scans. RESULTS: A total of 248 VLBW infants had evidence of IVH (22% of all VLBW infants, mean (SD) gestational age 26.8 (2.6) weeks). A quarter of the infants exhibited PVD. Spontaneous arrest of PVD occurred without treatment in 38% of infants with PVD. Of the remaining 62% with persistent PVD, 48% received non-surgical treatment only (pharmacological and/or drainage of cerebrospinal fluid by serial lumbar punctures), 34% received surgical treatment with insertion of a ventriculoperitoneal reservoir and/or shunt, and 18% died. The development of PVD after IVH and adverse short term outcome, such as the requirement for surgery, were predicted most strongly by the severity of IVH. CONCLUSIONS: These data reflect the natural history of PVD in the 1990s and show that, despite a slight reduction in its overall incidence, there appears to be a more aggressive course, with appreciable mortality and morbidity in the extremely premature infant. The major predictor of adverse short term outcome, defined as death or need for surgical intervention, was the severity of IVH. These findings may be valuable for the management of very small premature infants. PMID- 12091290 TI - In vitro comparison of nasal continuous positive airway pressure devices for neonates. AB - OBJECTIVE: To compare the resistance in vitro of different devices used for the delivery of nasal continuous positive airway pressure (NCPAP) in neonates. DESIGN: Flows of 4-8 litres/min were passed through a selection of neonatal NCPAP devices (single prong, Duotube, Argyle prong, Hudson prong, Infant Flow Driver), and the resultant fall in pressure measured using a calibrated pressure transducer. RESULTS: The decrease in pressure (cm H(2)O) for each device (size in parentheses) at a constant flow of 6 litres/min was: Duotube: (2.5), 21; (3.0), 6.2; (3.5), 2.3; single prong: (2.5), 4.4; (3.0), 2.1; (3.5), 1.2; Argyle prong: (XS), 3.6; (S), 1.9; (L), 1.5; Hudson prong: (0), 3.1; (1), 1.8; (2), 0.6; (3), 0.4; (4), 0.3; Infant Flow Driver: (small), 0.3; (medium), -0.3; (large), -0.5. CONCLUSIONS: A large variation in the potential fall in pressure may occur in the clinical setting. Devices with short double prongs had the lowest resistance to flow. These results have implications in the selection of the optimal device/s for clinical application and for future comparisons in randomised trials of NCPAP in neonates. PMID- 12091291 TI - Plasma glucose levels in term infants who are appropriate size for gestation and exclusively breast fed. AB - AIM: To evaluate the plasma glucose levels in normal, term infants who were appropriate size for gestational age (AGA) and exclusively breast fed, and to assess the influence of parity of the mother, mode of delivery, and time of feed on the glucose levels. METHOD: A total of 200 healthy, term, AGA infants were longitudinally evaluated at 3, 6, 24, and 72 hours of life. Plasma glucose was estimated from heel prick capillary samples. The influence of mode of delivery, parity, and interval between feeds on plasma glucose was analysed. RESULTS: There was no significant difference between the plasma glucose levels of the cohorts at any of the sampling time points. Parity, mode of delivery, and time since the last feed did not affect plasma glucose. Satisfactory glucose levels were maintained even when infants remained unfed up to 6 hours of age. Infants with plasma glucose concentrations less than 2.2 mmol/l at 3 hours of age were more likely to have low sugar concentration (< 2.5 mmol/l) at 72 hours (RR = 6.55, 95% confidence interval 3.93 to 10.92). CONCLUSIONS: A term, breast fed infant may have its own distinct plasma glucose levels, showing no significant variation between 3 and 72 hours of age. Plasma glucose levels are satisfactorily maintained in normal term infants without resort to prelacteal feeds. Mode of delivery, parity of the mother, and interval between feeds did not influence plasma glucose. Biochemical thresholds for hypoglycaemia do not seem to be of practical importance in asymptomatic, normal, term, breast fed infants. PMID- 12091292 TI - Zinc protoporphyrin/haem ratio and plasma ferritin in preterm infants. AB - OBJECTIVE: To study the utility of the zinc protoporphyrin/haem (ZPP/H) ratio as a measure of iron status in healthy, growing, preterm infants. METHOD: ZPP/H was measured in 109 well, preterm infants from the time of hospital discharge until 1 year of age (637 determinations). RESULTS: ZPP/H was initially high, but steadily declined. This was opposite to what was expected from the known changes in iron stores during the first year of life and the observed changes in plasma ferritin. Subjects with higher ZPP/H ratios tended to have lower ferritins, but changes in ZPP/H in a given subject were poorly reflected by changes in plasma ferritin. Between 6 and 9 months of age, ZPP/H correlated with other measures of iron status, but serum ferritin concentration did not. CONCLUSION: Use of the ZPP/H ratio as a measure of iron status during the first year of life appears to be confounded by the developmental changes in ZPP/H, but in the later half of this period it may be a better measure of iron status than serum ferritin. PMID- 12091293 TI - Multidrug resistant neonatal sepsis in Peshawar, Pakistan. AB - OBJECTIVE: To investigate the spectrum of organisms causing neonatal sepsis in Peshawar, Pakistan and to assess their sensitivity to various groups of drugs. METHODS: Blood taken from newborn babies admitted to the special care baby unit at the Khyber Teaching Hospital with a clinical diagnosis of neonatal sepsis was cultured. The data obtained from October 1997 to December 2000 were analysed and the results tabulated. RESULTS: A total of 1598 blood cultures were taken; 1003 were positive (positivity rate 62.8%). Escherichia coli was the most common organism found (36.6%), followed by Staphylococcus aureus (29.5%), Pseudomonas (22.4%), Klebsiella (7.6%), and Proteus (3.8%). No group B streptococcus was grown. Listeria monocytogenes was found in one cerebrospinal fluid culture. E coli and Pseudomonas showed a high degree of resistance to commonly used antibiotics (ampicillin, augmentin, and gentamicin), a moderate degree of resistance to cephalosporin (cefotaxime, ceftzidime, and ceftrioxone), and low resistance to drugs not used for newborn babies (ofloxacin, ciprofloxacin, and enoxabid). S aureus showed a low resistance to all three groups of antibiotics. CONCLUSION: Neonatal sepsis remains one of the leading causes of neonatal admission, morbidity, and mortality in developing countries. Gram negative organisms are the major cause of neonatal sepsis in Peshawar. Such organisms have developed multidrug resistance, and management of patients infected with them is becoming a problem in developing countries. PMID- 12091294 TI - A three year follow up of preterm infants after moderately early treatment with dexamethasone. AB - OBJECTIVE: To assess the effect of moderately early postnatal dexamethasone treatment on growth and neurodevelopmental outcome in preterm infants. METHODS: Thirty preterm infants enrolled in a randomised clinical trial to investigate the effectiveness of moderately early dexamethasone administration in the treatment of chronic lung disease were routinely followed up. Fifteen babies received a total dose of 4.75 mg/kg over 14 days from the 10th day of life, and 15 babies were untreated. Five infants in each group received open label steroids to facilitate extubation later in their clinical course. Growth and neurodevelopmental outcome are reported. RESULTS: The mean body weight, height, and head circumference as well as the number of babies with anthropometric measurements within normal range were similar in treated and untreated babies. There was no significant difference between treated and control groups with respect to incidence of cerebral palsy, major neurosensory impairment, mean intelligence quotient scores, and behavioural abnormalities. CONCLUSIONS: Postnatal dexamethasone treatment with the schedule used in this study did not impair growth and neurodevelopmental outcome in preterm infants. Data from larger trials have raised major concern that postnatal steroid treatment may increase neurodevelopmental impairment. The full extent of the risk will only be known when more trials have reported follow up data. PMID- 12091295 TI - Growth effects of systemic versus inhaled steroids in chronic lung disease. AB - AIM: To compare the effects of inhaled and systemic steroids on growth in very low birthweight (VLBW) infants with chronic lung disease (CLD). METHODS: Sixteen babies with CLD randomly received inhaled budesonide (100 microg four times daily for 10 days via Aerochamber) or systemic steroids (dexamethasone 0.5 mg/kg/day, reducing over nine days). Linear growth (lower leg length, LLL) was measured by knemometry twice weekly. RESULTS: The gestational age, birth weight, postnatal age, and LLL velocity (LLLvel) were similar between the two groups at the start of treatment. At the end of the treatment period, LLLvel was reduced in the dexamethasone group (mean -0.01 mm/day) but had increased in the budesonide group (mean 0.48 mm/day). Mean weight gain was non-significantly lower in the dexamethasone group (5.8 g/kg/day) compared to the budesonide group (mean 12.7 g/kg/day). CONCLUSION: Inhaled budesonide has less short term effects on growth than systemically administered dexamethasone. PMID- 12091296 TI - Randomised double blind placebo controlled trial of inhaled fluticasone propionate in infants with chronic lung disease. AB - In a double blind randomised controlled trial, 30 infants with chronic lung disease received fluticasone propionate or placebo for one year. There were no significant differences between treatment groups in the incidence of any day or night time symptoms or any other outcome measures. PMID- 12091297 TI - Percutaneous endovascular catheter aspiration thrombectomy of severe superior vena cava syndrome. AB - We report a combined percutaneous endovascular approach, including thrombus aspiration and catheter directed local thrombolysis, followed by systemic thrombolytic therapy to treat severe superior vena cava syndrome in a 2 and 1/2 week old infant. This procedure was performed on the fifth postoperative day after major surgery. No treatment complications were observed. The only predisposing condition for thrombosis was a central venous line. No other acquired or genetic risk factors for thrombosis could be found. PMID- 12091298 TI - Dr William Farr of Shropshire (1807-1883): obstetric mortality and training. AB - William Farr, chief statistician to the General Register Office for more than 40 years, was the most significant medical epidemiologist and statistician of the Victorian era. Often working behind the scenes, he helped to bring about many advances in hygiene and public health as well as developing a modern approach to the classification of disease and the collection and analysis of medical information data. PMID- 12091299 TI - Familial spontaneous pneumothorax. PMID- 12091300 TI - parachute/n-cadherin is required for morphogenesis and maintained integrity of the zebrafish neural tube. AB - N-cadherin (Ncad) is a classical cadherin that is implicated in several aspects of vertebrate embryonic development, including somitogenesis, heart morphogenesis, neural tube formation and establishment of left-right asymmetry. However, genetic in vivo analyses of its role during neural development have been rather limited. We report the isolation and characterization of the zebrafish parachute (pac) mutations. By mapping and candidate gene analysis, we demonstrate that pac corresponds to a zebrafish n-cadherin (ncad) homolog. Three mutant alleles were sequenced and each is likely to encode a non-functional Ncad protein. All result in a similar neural tube phenotype that is most prominent in the midbrain, hindbrain and the posterior spinal cord. Neuroectodermal cell adhesion is altered, and convergent cell movements during neurulation are severely compromised. In addition, many neurons become progressively displaced along the dorsoventral and the anteroposterior axes. At the cellular level, loss of Ncad affects beta-catenin stabilization/localization and causes mispositioned and increased mitoses in the dorsal midbrain and hindbrain, a phenotype later correlated with enhanced apoptosis and the appearance of ectopic neurons in these areas. Our results thus highlight novel and crucial in vivo roles for Ncad in the control of cell convergence, maintenance of neuronal positioning and dorsal cell proliferation during vertebrate neural tube development. PMID- 12091301 TI - Gliogenesis in Drosophila: genome-wide analysis of downstream genes of glial cells missing in the embryonic nervous system. AB - In Drosophila, the glial cells missing (gcm) gene encodes a transcription factor that controls the determination of glial versus neuronal fate. In gcm mutants, presumptive glial cells are transformed into neurons and, conversely, when gcm is ectopically misexpressed, presumptive neurons become glia. Although gcm is thought to initiate glial cell development through its action on downstream genes that execute the glial differentiation program, little is known about the identity of these genes. To identify gcm downstream genes in a comprehensive manner, we used genome-wide oligonucleotide arrays to analyze differential gene expression in wild-type embryos versus embryos in which gcm is misexpressed throughout the neuroectoderm. Transcripts were analyzed at two defined temporal windows during embryogenesis. During the first period of initial gcm action on determination of glial cell precursors, over 400 genes were differentially regulated. Among these are numerous genes that encode other transcription factors, which underscores the master regulatory role of gcm in gliogenesis. During a second later period, when glial cells had already differentiated, over 1200 genes were differentially regulated. Most of these genes, including many genes for chromatin remodeling factors and cell cycle regulators, were not differentially expressed at the early stage, indicating that the genetic control of glial fate determination is largely different from that involved in maintenance of differentiated cells. At both stages, glial-specific genes were upregulated and neuron-specific genes were downregulated, supporting a model whereby gcm promotes glial development by activating glial genes, while simultaneously repressing neuronal genes. In addition, at both stages, numerous genes that were not previously known to be involved in glial development were differentially regulated and, thus, identified as potential new downstream targets of gcm. For a subset of the differentially regulated genes, tissue specific in vivo expression data were obtained that confirmed the transcript profiling results. This first genome-wide analysis of gene expression events downstream of a key developmental transcription factor presents a novel level of insight into the repertoire of genes that initiate and maintain cell fate choices in CNS development. PMID- 12091302 TI - The zebrafish T-box genes no tail and spadetail are required for development of trunk and tail mesoderm and medial floor plate. AB - T-box genes encode transcriptional regulators that control many aspects of embryonic development. Here, we demonstrate that the mesodermally expressed zebrafish spadetail (spt)/VegT and no tail (ntl)/Brachyury T-box genes are semi redundantly and cell-autonomously required for formation of all trunk and tail mesoderm. Despite the lack of posterior mesoderm in spt(-);ntl(-) embryos, dorsal ventral neural tube patterning is relatively normal, with the notable exception that posterior medial floor plate is completely absent. This contrasts sharply with observations in single mutants, as mutations singly in ntl or spt enhance posterior medial floor plate development. We find that ntl function is required to repress medial floor plate and promote notochord fate in cells of the wild type notochord domain and that spt and ntl together are required non cell autonomously for medial floor plate formation, suggesting that an inducing signal present in wild-type mesoderm is lacking in spt(-);ntl(-) embryos. PMID- 12091303 TI - A common translational control mechanism functions in axial patterning and neuroendocrine signaling in Drosophila. AB - Translational repression of maternal nanos (nos) mRNA by a cis-acting Translational Control Element (TCE) in the nos 3'UTR is critical for anterior posterior patterning of the Drosophila embryo. We show, through ectopic expression experiments, that the nos TCE is capable of repressing gene expression at later stages of development in neuronal cells that regulate the molting cycle. Our results predict additional targets of TCE-mediated repression within the nervous system. They also suggest that mechanisms that regulate maternal mRNAs, like TCE-mediated repression, may function more widely during development to spatially or temporally control gene expression. PMID- 12091304 TI - The zinc finger protein REF-2 functions with the Hox genes to inhibit cell fusion in the ventral epidermis of C. elegans. AB - During larval development in C. elegans, some of the cells of the ventral epidermis, the Pn.p cells, fuse with the growing epidermal syncytium hyp7. The pattern of these cell fusions is regulated in a complex, sexually dimorphic manner. It is essential that some Pn.p cells remain unfused in order for some sex specific mating structures to be generated. The pattern of Pn.p cell fusion is regulated combinatorially by two genes of the C. elegans Hox gene cluster: lin-39 and mab-5. Some of the complexity in the Pn.p cell fusion pattern arises because these two Hox proteins can regulate each other's activities. We describe a zinc finger transcription factor, REF-2, that is required for the Pn.p cells to be generated and to remain unfused. REF-2 functions with the Hox proteins to prevent Pn.p cell fusion. ref-2 may also be a transcriptional target of the Hox proteins. PMID- 12091305 TI - Neural crest progenitors of the melanocyte lineage: coat colour patterns revisited. AB - Neural crest-derived melanoblasts are the progenitors of melanocytes, the pigment cells of the skin, hair and choroid. Previous studies of adult chimaeric mice carrying different coat colour markers have suggested that the total melanocyte population is derived from a small number of melanoblast progenitors, each of which generates a discrete unilateral transverse band of colour. This work also suggested minimal mixing of cells between clones. We have used two complementary approaches to assess the behaviour of migrating clones of melanoblasts directly in the developing embryo. First, we made aggregation chimaeras between transgenic Dct-lacZ and non-transgenic embryos, in which lacZ is a marker for melanoblasts. Second, we generated transgenic mice carrying a modified lacZ reporter construct containing a 289 base pair duplication (laacZ) under the control of the Dct promoter. The laacZ transgene is normally inactive, but reverts to wild-type lacZ at low frequency, labelling a cell and all of its progeny at random. Mosaic embryos containing labelled melanoblast clones were generated. In contrast to previous data, chimaeric and mosaic embryonic melanoblast patterns suggest that: (1) there is a large number of melanoblast progenitors; (2) there is a pool of melanoblasts in the cervical region; (3) different cell dispersion mechanisms may operate in the head and trunk regions; and (4) there is extensive axial mixing between clones. PMID- 12091306 TI - Transmeiotic differentiation of zebrafish germ cells into functional sperm in culture. AB - Because cell culture systems are easily accessible for experimental genetic manipulation, male germ cell culture is of great usefulness in creating sperm vectors. This report describes that cultured male germ cells of zebrafish (Danio rerio) underwent mitosis and transmeiotic differentiation, including the entire process of meiosis, to develop into functional sperm. Enzymatically dissociated testicular cells containing germ cells were co-cultured on feeder cells derived from tumor-like testis, which exhibited features characteristic of Sertoli cells such as phagocytic activity and transcription of the Wilms' tumor suppressor wt1 and sox9a genes. Germ cells formed a clump, divided by mitosis, and differentiated into flagellated sperm on the feeders. Expression of the germ cell marker gene vas was prolonged in co-culture with the feeders, compared with culture of dissociated testicular cells alone, indicating that the feeder cells stimulate proliferation of spermatogonia. When cultured germ cells/sperm with the feeders were used for in vitro fertilization, normal embryos were obtained. Addition of the thymidine analogue 5-bromo-2'-deoxyuridine (BrdU) into culture medium resulted in BrdU-positive sperm and four-cell stage embryos after in vitro fertilization. This culture system should prove useful not only in producing transfected functional sperm, but also in analyzing the regulatory function of testicular somatic cells on the mitosis and meiosis of male germ cells in vertebrates. PMID- 12091307 TI - unc-53 controls longitudinal migration in C. elegans. AB - Cell migration and outgrowth are thought to be based on analogous mechanisms that require repeated cycles of process extension, reading and integration of multiple directional signals, followed by stabilisation in a preferred direction, and renewed extension. We have characterised a C. elegans gene, unc-53, that appears to act cell autonomously in the migration and outgrowth of muscles, axons and excretory canals. Abrogation of unc-53 function disrupts anteroposterior outgrowth in those cells that normally express the gene. Conversely, overexpression of unc-53 in bodywall muscles leads to exaggerated outgrowth. UNC 53 is a novel protein conserved in vertebrates that contains putative SH3- and actin-binding sites. unc-53 interacts genetically with sem-5 and we demonstrated a direct interaction in vitro between UNC-53 and the SH2-SH3 adaptor protein SEM 5/GRB2. Thus, unc-53 is involved in longitudinal navigation and might act by linking extracellular guidance cues to the intracellular cytoskeleton. PMID- 12091308 TI - Integrins regulate DLG/FAS2 via a CaM kinase II-dependent pathway to mediate synapse elaboration and stabilization during postembryonic development. AB - Calcium/calmodulin dependent kinase II (CaMKII), PDZ-domain scaffolding protein Discs-large (DLG), immunoglobin superfamily cell adhesion molecule Fasciclin 2 (FAS2) and the position specific (PS) integrin receptors, including betaPS and its alpha partners (alphaPS1, alphaPS2, alphaPS3/alphaVolado), are all known to regulate the postembryonic development of synaptic terminal arborization at the Drosophila neuromuscular junction (NMJ). Recent work has shown that DLG and FAS2 function together to modulate activity-dependent synaptic development and that this role is regulated by activation of CaMKII. We show that PS integrins function upstream of CaMKII in the development of synaptic architecture at the NMJ. betaPS integrin physically associates with the synaptic complex anchored by the DLG scaffolding protein, which contains CaMKII and FAS2. We demonstrate an alteration of the FAS2 molecular cascade in integrin regulatory mutants, as a result of CaMKII/integrin interactions. Regulatory betaPS integrin mutations increase the expression and synaptic localization of FAS2. Synaptic structural defects in betaPS integrin mutants are rescued by transgenic overexpression of CaMKII (proximal in pathway) or genetic reduction of FAS2 (distal in pathway). These studies demonstrate that betaPS integrins act through CaMKII activation to control the localization of synaptic proteins involved in the development of NMJ synaptic morphology. PMID- 12091310 TI - The induced sector Arabidopsis apical embryonic fate map. AB - Creation of an embryonic fate map may provide insight into the patterns of cell division and specification contributing to the apical region of the early Arabidopsis embryo. A fate map has been constructed by inducing genetic chimerism during the two-apical-cell stage of embryogenesis to determine if the orientation of the first anticlinal cell division correlates with later developmental axes. Chimeras were also used to map the relative locations of precursors of the cotyledon and leaf primordia. Genetic chimeras were induced in embryos doubly heterozygous for a heat shock regulated Cre recombinase and a constitutively expressed beta-glucuronidase (GUS) gene flanked by the loxP binding sites for Cre. Individual cells in the two-apical-cell stage embryo responding to heat shock produce GUS-negative daughter cells. Mature plants grown from seed derived from treated embryos were scored for GUS-negative sector extent in the cotyledons and leaves. The GUS-negative daughters of apical cells had a strong tendency to contribute primarily to one cotyledon or the other and to physically adjacent true leaf margins. This result indicated that patterns of early cell division correlate with later axes of symmetry in the embryo and that these patterns partially limit the fates available for adoption by daughter cells. However, GUS negative sectors were shared between all regions of the mature plant, suggesting that there is no strict fate restriction imposed on the daughters of the first apical cells. PMID- 12091309 TI - Wingless blocks bristle formation and morphogenetic furrow progression in the eye through repression of Daughterless. AB - In the developing eye, wingless activity represses proneural gene expression (and thus interommatidial bristle formation) and positions the morphogenetic furrow by blocking its initiation in the dorsal and ventral regions of the presumptive eye. We provide evidence that wingless mediates both effects, at least in part, through repression of the basic helix-loop-helix protein Daughterless. daughterless is required for high proneural gene expression and furrow progression. Ectopic expression of wingless blocks Daughterless expression in the proneural clusters. This repression, and that of furrow progression, can be mimicked by an activated form of armadillo and blocked by a dominant negative form of pangolin/TCF. Placing daughterless under the control of a heterologous promoter blocks the ability of ectopic wingless to inhibit bristle formation and furrow progression. hedgehog and decapentapleigic could not rescue the wingless furrow progression block, indicating that wingless acts downstream of these genes. In contrast, Atonal and Scute, which are thought to heterodimerize with Daughterless to promote furrow progression and bristle formation, respectively, can block ectopic wingless action. These results are summarized in a model where daughterless is a major, but probably not the only, target of wingless action in the eye. PMID- 12091311 TI - Linear signaling in the Toll-Dorsal pathway of Drosophila: activated Pelle kinase specifies all threshold outputs of gene expression while the bHLH protein Twist specifies a subset. AB - Differential activation of the Toll receptor leads to the formation of a broad Dorsal nuclear gradient that specifies at least three patterning thresholds of gene activity along the dorsoventral axis of precellular embryos. We investigate the activities of the Pelle kinase and Twist basic helix-loop-helix (bHLH) transcription factor in transducing Toll signaling. Pelle functions downstream of Toll to release Dorsal from the Cactus inhibitor. Twist is an immediate-early gene that is activated upon entry of Dorsal into nuclei. Transgenes misexpressing Pelle and Twist were introduced into different mutant backgrounds and the patterning activities were visualized using various target genes that respond to different thresholds of Toll-Dorsal signaling. These studies suggest that an anteroposterior gradient of Pelle kinase activity is sufficient to generate all known Toll-Dorsal patterning thresholds and that Twist can function as a gradient morphogen to establish at least two distinct dorsoventral patterning thresholds. We discuss how the Dorsal gradient system can be modified during metazoan evolution and conclude that Dorsal-Twist interactions are distinct from the interplay between Bicoid and Hunchback, which pattern the anteroposterior axis. PMID- 12091312 TI - BMP2 is a positive regulator of Nodal signaling during left-right axis formation in the chicken embryo. AB - A model of left-right axis formation in the chick involves inhibition of bone morphogenetic proteins by the antagonist Car as a mechanism of upregulating Nodal in the left lateral plate mesoderm. By contrast, expression of CFC, a competence factor, which is absolutely required for Nodal signaling in the lateral plate mesoderm is dependent on a functional BMP signaling pathway. We have therefore investigated the relationship between BMP and Nodal in further detail. We implanted BMP2 and Noggin-expressing cells into the left lateral plate and paraxial mesoderm and observed a strong upregulation of Nodal and its target genes Pitx2 and Nkx3.2. In addition Cfc, the Nodal type II receptor ActrIIa and Snr were found to depend on BMP signaling for their expression. Comparison of the expression domains of Nodal, Bmp2, Car and Cfc revealed co-expression of Nodal, Cfc and Bmp2, while Car and Nodal only partially overlapped. Ectopic application of BMP2, Nodal, and Car as well as combinations of this signaling molecules to the right lateral plate mesoderm revealed that BMP2 and Car need to synergize in order to specify left identity. We propose a novel model of left-right axis formation, which involves BMP as a positive regulator of Nodal signaling in the chick embryo. PMID- 12091313 TI - BMP signaling positively regulates Nodal expression during left right specification in the chick embryo. AB - Exogenous application of BMP to the lateral plate mesoderm (LPM) of chick embryos at the early somite stage had a positive effect on Nodal expression. BMP applications into the right LPM were followed by a rapid activation of Nodal, while applications into the left LPM resulted in expansion of the normal domain of Nodal expression. Conversely, blocking of BMP signaling by Noggin in the left LPM interfered with the activation of Nodal expression. These results support a positive role for endogenous BMP on Nodal expression in the LPM. We also report that BMP positively regulates the expression of Caronte, Snail and Cfc in both the left and right LPM. BMP-treated embryos had molecular impairment of the midline with downregulation of Lefty1, Brachyury and Shh but we also show that the midline defect was not sufficient to induce ectopic Nodal expression. We discuss our findings in the context of the known molecular control of the specification of left-right asymmetry. PMID- 12091314 TI - An essential role of a FoxD gene in notochord induction in Ciona embryos. AB - A key issue for understanding the early development of the chordate body plan is how the endoderm induces notochord formation. In the ascidian Ciona, nuclear accumulation of beta-catenin is the first step in the process of endoderm specification. We show that nuclear accumulation of beta-catenin directly activates the gene (Cs-FoxD) for a winged helix/forkhead transcription factor and that this gene is expressed transiently at the 16- and 32-cell stages in endodermal cells. The function of Cs-FoxD, however, is not associated with differentiation of the endoderm itself but is essential for notochord differentiation or induction. In addition, it is likely that the inductive signal that appears to act downstream of Cs-FoxD does not act over a long range. It has been suggested that FGF or Notch signal transduction pathway mediates ascidian notochord induction. Our previous study suggests that Cs-FGF4/6/9 is partially involved in the notochord induction. The present experimental results suggest that the expression and function of Cs-FGF4/6/9 and Cs-FoxD are not interdependent, and that the Notch pathway is involved in B-line notochord induction downstream of Cs-FoxD. PMID- 12091315 TI - The Foxh1-dependent autoregulatory enhancer controls the level of Nodal signals in the mouse embryo. AB - The TGFbeta-related growth factor Nodal governs anteroposterior (AP) and left right (LR) axis formation in the vertebrate embryo. A conserved intronic enhancer (ASE), containing binding sites for the fork head transcription factor Foxh1, modulates dynamic patterns of Nodal expression during early mouse development. This enhancer is responsible for early activation of Nodal expression in the epiblast and visceral endoderm, and at later stages governs asymmetric expression during LR axis formation. We demonstrate ASE activity is strictly Foxh1 dependent. Loss of this autoregulatory enhancer eliminates transcription in the visceral endoderm and decreases Nodal expression in the epiblast, but causes surprisingly discrete developmental abnormalities. Thus lowering the level of Nodal signaling in the epiblast disrupts both orientation of the AP axis and specification of the definitive endoderm. Targeted removal of the ASE also dramatically reduces left-sided Nodal expression, but the early events controlling LR axis specification are correctly initiated. However loss of the ASE disrupts Lefty2 (Leftb) expression and causes delayed Pitx2 expression leading to late onset, relatively minor LR patterning defects. The feedback loop is thus essential for maintenance of Nodal signals that selectively regulate target gene expression in a temporally and spatially controlled fashion in the mouse embryo. PMID- 12091316 TI - poirot, a new regulatory gene of Drosophila oskar acts at the level of the short Oskar protein isoform. AB - Embryonic germ cell formation and abdomen development in Drosophila requires localisation and site specific translation of oskar mRNA in the posterior part of the oocyte. Targeting of oskar function to the posterior pole of the oocyte needs a large set of proteins and RNAs, encoded by posterior group genes. Consequently, mutations in the posterior group genes can result in embryos without abdomens and/or germ cells. During a systematic hobo-mediated mutant isolation screen, we identified poirot, a novel posterior group gene, owing to its germ cell-less phenotype. We show that the lack of poirot activity dramatically decreases OSK protein levels, without affecting the oskar mRNA distribution. In poirot mutant oocytes, delocalised OSK protein is observed, indicating that wild-type poirot has a role in the anchoring process of the OSK protein at the posterior pole. Furthermore, we demonstrate that poirot acts in an isoform-specific manner, only the short OSK isoform is affected, while the long OSK isoform remains at wild type levels in poirot mutants. PMID- 12091317 TI - Absence of Cajal-Retzius cells and subplate neurons associated with defects of tangential cell migration from ganglionic eminence in Emx1/2 double mutant cerebral cortex. AB - Emx1 and Emx2, mouse orthologs of the Drosophila head gap gene, ems, are expressed during corticogenesis. Emx2 null mutants exhibit mild defects in cortical lamination. Segregation of differentiating neurons from proliferative cells is normal for the most part, however, reelin-positive Cajal-Retzius cells are lost by the late embryonic period. Additionally, late-born cortical plate neurons display abnormal position. These types of lamination defects are subtle in the Emx1 mutant cortex. In the present study we show that Emx1 and Emx2 double mutant neocortex is much more severely affected. Thickness of the cerebral wall was diminished with the decrease in cell number. Bromodeoxyuridine uptake in the germinal zone was nearly normal; moreover, no apparent increase in cell death or tetraploid cell number was observed. However, tangential migration of cells from the ganglionic eminence into the neocortex was greatly inhibited. The wild-type ganglionic eminence cells transplanted into Emx1/2-double mutant telencephalon did not move to the cortex. MAP2-positive neuronal bodies and RC2-positive radial glial cells emerged normally, but the laminar structure subsequently formed was completely abnormal. Furthermore, both corticofugal and corticopetal fibers were predominantly absent in the cortex. Most importantly, neither Cajal-Retzius cells nor subplate neurons were found throughout E11.5-E18.5. Thus, this investigation suggests that laminar organization in the cortex or the production of Cajal Retzius cells and subplate neurons is interrelated to the tangential movement of cells from the ganglionic eminence under the control of Emx1 and Emx2. PMID- 12091318 TI - Planar cell polarization requires Widerborst, a B' regulatory subunit of protein phosphatase 2A. AB - We have identified widerborst (wdb), a B' regulatory subunit of PP2A, as a conserved component of planar cell polarization mechanisms in both Drosophila and in zebrafish. In Drosophila, wdb acts at two steps during planar polarization of wing epithelial cells. It is required to organize tissue polarity proteins into proximal and distal cortical domains, thus determining wing hair orientation. It is also needed to generate the polarized membrane outgrowth that becomes the wing hair. Widerborst activates the catalytic subunit of PP2A and localizes to the distal side of a planar microtubule web that lies at the level of apical cell junctions. This suggests that polarized PP2A activation along the planar microtubule web is important for planar polarization. In zebrafish, two wdb homologs are required for convergent extension during gastrulation, supporting the conjecture that Drosophila planar cell polarization and vertebrate gastrulation movements are regulated by similar mechanisms. PMID- 12091319 TI - Removal of dystroglycan causes severe muscular dystrophy in zebrafish embryos. AB - Muscular dystrophy is frequently caused by disruption of the dystrophin glycoprotein complex (DGC), which links muscle cells to the extracellular matrix. Dystroglycan, a central component of the DGC, serves as a laminin receptor via its extracellular alpha subunit, and interacts with dystrophin (and thus the actin cytoskeleton) through its integral membrane beta subunit. We have removed the function of dystroglycan in zebrafish embryos. In contrast to mouse, where dystroglycan mutations lead to peri-implantation lethality, dystroglycan is dispensable for basement membrane formation during early zebrafish development. At later stages, however, loss of dystroglycan leads to a disruption of the DGC, concurrent with loss of muscle integrity and necrosis. In addition, we find that loss of the DGC leads to loss of sarcomere and sarcoplasmic reticulum organisation. The DGC is required for long-term survival of muscle cells in zebrafish, but is dispensable for muscle formation. Dystroglycan or the DGC is also required for normal sarcomere and sarcoplasmic reticulum organisation. Because zebrafish embryos lacking dystroglycan share several characteristics with human muscular dystrophy, they should serve as a useful model for the disease. In addition, knowing the dystroglycan null phenotype in zebrafish will facilitate the isolation of other molecules involved in muscular dystrophy pathogenesis. PMID- 12091320 TI - PTEN is essential for cell migration but not for fate determination and tumourigenesis in the cerebellum. AB - PTEN is a tumour suppressor gene involved in cell cycle control, apoptosis and mediation of adhesion and migration signalling. Germline mutations of PTEN in humans are associated with familial tumour syndromes, among them Cowden disease. Glioblastomas, highly malignant glial tumours of the central nervous system frequently show loss of PTEN. Recent reports have outlined some aspects of PTEN function in central nervous system development. Using a conditional gene disruption approach, we inactivated Pten in mice early during embryogenesis locally in a region specific fashion and later during postnatal development in a cell-specific manner, to study the role of PTEN in differentiation, migration and neoplastic transformation. We show that PTEN is required for the realisation of normal cerebellar architecture, for regulation of cell and organ size, and for proper neuronal and glial migration. However, PTEN is not required for cell differentiation and lack of PTEN is not sufficient to induce neoplastic transformation of neuronal or glial cells PMID- 12091321 TI - Hearing loss caused by progressive degeneration of cochlear hair cells in mice deficient for the Barhl1 homeobox gene. AB - The cochlea of the mammalian inner ear contains three rows of outer hair cells and a single row of inner hair cells. These hair cell receptors reside in the organ of Corti and function to transduce mechanical stimuli into electrical signals that mediate hearing. To date, the molecular mechanisms underlying the maintenance of these delicate sensory hair cells are unknown. We report that targeted disruption of Barhl1, a mouse homolog of the Drosophila BarH homeobox genes, results in severe to profound hearing loss, providing a unique model for the study of age-related human deafness disorders. Barhl1 is expressed in all sensory hair cells during inner ear development, 2 days after the onset of hair cell generation. Loss of Barhl1 function in mice results in age-related progressive degeneration of both outer and inner hair cells in the organ of Corti, following two reciprocal longitudinal gradients. Our data together indicate an essential role for Barhl1 in the long-term maintenance of cochlear hair cells, but not in the determination or differentiation of these cells. PMID- 12091325 TI - P-glycoprotein targeting: a unique strategy to selectively eliminate immunoreactive T cells. AB - T lymphocytes have been found to harbor P-glycoprotein (Pgp) and to demonstrate modulation of its ion channel transporter function according to the state of activation of T lymphocytes. We hypothesized that cytotoxic chemicals that are extruded by Pgp could be used to specifically eliminate immunoreactive T-cell populations. In this study, we evaluated the capacity of 4,5-dibromorhodamine methyl ester (TH9402), a photosensitizer structurally similar to rhodamine, a dye transported by Pgp, and which becomes highly cytotoxic on activation with visible light to selectively deplete alloreactive T lymphocytes. Stimulation of T cells with mitogens or allogeneic major histocompatibility complex-mismatched cells resulted in the preferential retention of the TH9402 rhodamine-derivative in activated T cells, both CD4+ and CD8+. Photodynamic cell therapy of TH9402 exposed T cells led to the selective elimination of immunoreactive T-cell populations. In addition, this treatment preserved resting T cells and their capacity to respond to third-party cells. Inhibition of Pgp enhanced cellular trapping of the dye in nonactivated T cells and resulted in their depletion after exposure to light. Targeting of Pgp-deficient cells may therefore represent an appealing strategy for the prevention and treatment of graft-versus-host disease and other alloimmune or autoimmune disorders. PMID- 12091326 TI - Characterization of a new subpopulation of mouse CD8alpha+ B220+ dendritic cells endowed with type 1 interferon production capacity and tolerogenic potential. AB - We describe a new B220+ subpopulation of immaturelike dendritic cells (B220+ DCs) with low levels of expression of major histocompatibility complex (MHC) and costimulatory molecules and markedly reduced T-cell stimulatory potential, located in the thymus, bone marrow, spleen, and lymph nodes. B220+ DCs display ultrastructural characteristics resembling those of human plasmacytoid cells and accordingly produce interferon-alpha after virus stimulation. B220+ DCs acquired a strong antigen-presenting cell capacity on incubation with CpG oligodeoxynucleotides, concomitant with a remarkable up-regulation of MHC and costimulatory molecules and the production of interleukin-12 (IL-12) and IL-10. Importantly, our data suggest that nonstimulated B220+ DCs represent a subset of physiological tolerogenic DCs endowed with the capacity to induce a nonanergic state of T-cell unresponsiveness, involving the differentiation of T regulatory cells capable of suppressing antigen-specific T-cell proliferation. In conclusion, our data support the hypothesis that B220+ DCs represent a lymphoid organ subset of immature DCs with a dual role in the immune system-exerting a tolerogenic function in steady state but differentiating on microbial stimulation into potent antigen-presenting cells with type 1 interferon production capacity. PMID- 12091327 TI - Lamivudine therapy for prevention of immunosuppressive-induced hepatitis B virus reactivation in hepatitis B surface antigen carriers. AB - Viral reactivation in hepatitis B surface antigen (HBsAg) carriers undergoing immunosuppressive therapy is well documented. To evaluate the role of lamivudine prophylaxis in Hepatitis B virus (HBV) carriers treated with immunosuppression for nonhepatic disorders, we reviewed our experience between 1997 and 2000 at Hadassah University Hospital (Jerusalem, Israel). Controls were patients who were HBV carriers and who, between 1990 and 1995, were treated for hematological malignancies but were not treated with lamivudine. Eighteen HBsAg-positive patients were treated with immunosuppression. Fourteen were males, with a mean age of 48 years. Eleven patients had lymphoma; 2 had colonic adenocarcinoma; and 5 had cryoglobulinemia, enophthalmitis, vasculitis, malignant histocytosis, or ulcerative colitis. Fourteen patients were treated with chemotherapy, and 4 with prolonged high-dose corticosteroids. All patients were HBsAg-positive; 4 had hepatitis B e antigen, and 10 had HBV DNA by polymerase chain reaction. Lamivudine was administered to 13 patients in the treatment group 1 to 60 days (mean, 15 days) before immunosuppressive treatment and continued 0.5 to 24 months (mean, 7 months) following initiation of immunosuppression. Mean follow-up after lamivudine administration was 21 months. Three patients died of lymphoma complications and 10 (77%) survived. None of the patients had clinical or serological evidence of HBV reactivation during or after lamivudine prophylaxis. Of 6 patients who presented with liver function test disturbances, 5 improved during combined lamivudine and immunosuppression treatment. At the end of follow up, HBV DNA became undetectable in 2 of 10 patients. In 2 patients, seroconversion from HBsAg to anti-HBs was observed. In contrast, 2 of 5 control patients had HBV reactivation. Lamivudine prophylaxis in HBsAg carriers receiving immunosuppressive therapy may prevent HBV reactivation and hepatic failure. PMID- 12091328 TI - Donor lymphocyte infusion for relapsed chronic myelogenous leukemia: prognostic relevance of the initial cell dose. AB - Donor lymphocyte infusion (DLI) can produce durable remissions in patients with chronic myelogenous leukemia (CML) who have a relapse after an allogeneic stem cell transplantation. However, the best modality to administer DLI is still unclear. The effect of the initial cell dose (ICD; ie, mononuclear cells x 10(8)/kg received in the first instance) on outcome was retrospectively analyzed in 298 of 344 patients treated with DLI at 51 centers. Patients were classified into 3 groups according to the ICD: 98 in group A ( 2.0). Additional infusions were given to 62%, 20%, and 5% of patients in groups A, B, and C, respectively. A lower ICD was associated with less graft-versus-host disease (GVHD; A, 26%; B, 53%; C, 62%; P <.001), less myelosuppression (A, 10%; B, 23%; C, 24%; P =.01), and similar response rate (A, 78%; B, 73%; C, 70%; P =.48). Nonadjusted estimates of 3-year survival, failure-free survival, and DLI-related mortality were 84%, 66%, and 5% respectively, in group A; 63%, 57%, and 20% in group B; and 58%, 45%, and 22% in group C. Outcome analysis was adjusted for patient age, donor type, sex of donor, sex mismatch, disease phase at transplantation, T-cell depletion, interval from transplantation to DLI, GVHD prior to relapse, relapse type, and date of DLI. After adjustment, lower ICD was associated with less GVHD, less myelosuppression, same response rate, better survival, better failure-free survival, and less DLI related mortality. Our results suggest that the first DLI dose should not exceed 0.2 x 10(8) mononuclear cells/kg. PMID- 12091329 TI - Severity of chronic graft-versus-host disease: association with treatment-related mortality and relapse. AB - Chronic graft-versus-host disease (cGVHD) is the leading cause of late treatment related deaths among recipients of allogeneic bone marrow and blood transplants. However, cGVHD is also associated with fewer relapses. We sought to determine whether severity of cGVHD predicts the magnitude of these effects. One impediment to such an analysis is the current limited/extensive grading system for cGVHD because this classification was designed to identify patients likely to benefit from systemic immune suppression and does not capture the severity of multiorgan involvement. We, therefore, first developed a grading system predictive for survival by using data from 1827 HLA-matched sibling allotransplant recipients reported to the International Bone Marrow Transplant Registry (IBMTR). We found Karnofsky performance score, diarrhea, weight loss, and cutaneous and oral involvement to be independent prognostic variables, from which we generated a grading scheme. We tested this scheme, the limited/extensive classification system, and a classification based on clinical impression of overall cGVHD severity (mild/moderate/severe) in parallel analyses of 1092 HLA-matched sibling transplant recipients from the IBMTR and 553 recipients of unrelated donor marrow from the National Marrow Donor Program. Presence of cGVHD was associated with fewer relapses (relative risk [RR], 0.5-0.6) but more treatment-related mortality (RR, 1.8-2.8) in the 3 analyses. No grading scheme correlated cGVHD severity with relapse rates, but all schemes predicted treatment-related mortality. Survival and disease-free survival of the most favorable cGVHD group in each scheme were similar, or better, than those of patients without cGVHD; these patients may not need aggressive or extended immune suppression. PMID- 12091330 TI - Comparison of chronic graft-versus-host disease after transplantation of peripheral blood stem cells versus bone marrow in allogeneic recipients: long term follow-up of a randomized trial. AB - In a previous multicenter phase III trial comparing peripheral blood stem cell transplantation (PBSCT) to bone marrow transplantation (BMT) from HLA-matched related donors, we found no statistically significant difference in the cumulative incidence of clinical extensive chronic graft-versus-host disease (GVHD) in the 2 groups. We have analyzed the results in more detail to determine whether the clinical characteristics of chronic GVHD after PBSCT might be distinct from those that occur after BMT. Clinical extensive chronic GVHD developed in 39 of 63 recipients of PBSCs and in 32 of 63 BM recipients who were alive and free of malignancy at day 100 after the transplantation. No significant differences were found in the time and type of onset of clinical extensive chronic GVHD or in the frequency of complications associated with severe morbidity. Involvement of skin and female genital tract was more frequent in PBSC recipients than in BM recipients. The cumulative incidence of chronic GVHD at 3 years was similar in the 2 groups, but the number of successive treatments needed to control chronic GVHD was higher after PBSCT than after BMT (P =.03), and the duration of glucocorticoid treatment was longer after PBSCT compared to BMT (P =.03). These results suggest that chronic GVHD after PBSCT may be more protracted and less responsive to current treatment than chronic GVHD after BMT. Assessment of the overall benefits of PBSCT compared to BMT will require continued long-term follow up of morbidity associated with chronic GVHD. PMID- 12091331 TI - Improved outcome in high-risk childhood acute lymphoblastic leukemia defined by prednisone-poor response treated with double Berlin-Frankfurt-Muenster protocol II. AB - One hundred ninety-eight children and adolescents were entered in the Associazione Italiana di Ematologia ed Oncologia Pediatrica (AIEOP)-ALL95 study for high-risk acute lymphoblastic leukemia (ALL). Inclusion criteria were poor response to initial prednisone/intrathecal methotrexate (prednisone-poor response [PPR]), resistance to induction therapy, translocation t(9;22), infants with the t(4;11), or CD10(-) ALL. The event-free survival (EFS) rate at 4 years was 56.5% (SE, 3.9%) for the entire group. The overall EFS rate in the current study was significantly better (P =.002) than that obtained in a comparable group of patients treated in the early 1990s in the AIEOP-ALL91 study. In particular, patients with PPR had a 4-year EFS of 61.1% (SE, 4.4%) versus 42.8% (SE, 5.4%) in the ALL 91 study (P =.008). Among PPR patients, those who were PPR-only (60.1%) that is, they achieved CR and were negative for t(9;22) and t(4;11) translocations-had the best outcomes with this intensive treatment, even when additional adverse features (hyperleukocytosis, T phenotype) were present (4-year EFS, 70.1%; SE, 4.7%). We attribute this improvement to the replacement of 6 alternating blocks of non-cross-resistant drugs with an 8-drug reinduction regimen (Berlin-Frankfurt-Muenster [BFM] protocol II), repeated twice, in the context of a standard BFM-type intensive chemotherapy for high-risk ALL. This modified therapy may lead to high cure rates for patients defined as at high risk for intrinsic resistance to corticosteroids only. PMID- 12091332 TI - Acute myeloid leukemia and myelodysplastic syndrome in children treated for cancer: comparison with primary presentation. AB - There has not been a reported series of children with therapy-induced myelodysplastic syndrome/acute myeloid leukemia (tMDS/tAML) who were treated systematically. This paper describes 24 children with tMDS/tAML who were assigned randomly to standard- or intensive-timing induction on protocol CCG 2891. Presenting features and outcomes of those children were compared with those of 960 patients with de novo MDS (62 patients) or AML (898 patients). Children with tMDS/tAML were older at presentation (P =.015), had lower white blood cell counts (P =.01), and were more likely to have MDS (21% vs 7%) (P =.02) and trisomy 8 (P =.06). Fewer had hepatomegaly (P =.02), splenomegaly (P =.03), hepatosplenomegaly (P =.02), or classic AML translocations [t(8;21), t(15;17), 16q22; P =.02]. They had a poorer induction rate (50% vs 72%, P =.016), overall survival (26% vs 47% at 3 years, P =.007), and event-free survival (21% vs 39% at 3 years, P =.023). Disease-free survival after achieving remission was similar (45% vs 53%, P =.868). Children with tMDS/tAML who received intensive-timing induction had better outcomes than those who received standard-timing induction (overall survival 32% vs 0%, P =.54). In this study, the latency period to development of tMDS/tAML was the same for presumed alkylator-induced as for topoisomerase induced myeloid leukemia. The findings of this study confirm that most children with tMDS/tAML have disease resistant to current therapies. Standard-timing induction appears less effective for this population. PMID- 12091333 TI - Hematopathologic and cytogenetic findings in imatinib mesylate-treated chronic myelogenous leukemia patients: 14 months' experience. AB - Imatinib mesylate, an Abl kinase inhibitor, produces sustained complete hematologic responses (CHRs) in chronic myelogenous leukemia (CML) patients, but the sequence and timing of morphologic and cytogenetic changes in CML patients during prolonged imatinib mesylate treatment has not been described. In this report, we document sequential hematologic and bone marrow findings in 19 interferon-refractory/interferon-intolerant chronic phase CML patients on imatinib mesylate for at least 14 months. Patients treated at an effective oral dose (300 to 600 mg per day) were followed with peripheral blood (PB) counts, marrow examination, and cytogenetic studies at 0, 2, 5, 8, 11, and 14 months. By 2 months, 17 of 19 patients achieved CHR; 1 reached CHR by 5 months, and 1 at 11 months. Five of 19 patients developed cytopenias requiring treatment interruption and/or dose reduction, but all were able to continue in CHR on study. In contrast to interferon-alfa treatment, imatinib mesylate-treated CML patients achieved not only CHR but complete morphologic marrow response. Normalization of marrow lagged behind PB response; however, by 8 months, all marrows showed normal or reduced cellularity without morphologic evidence of CML. Eighteen of 19 patients continued in CHR and morphologic marrow remission at 14 months; 1 patient relapsed with chronic phase CML. Although hematologic and marrow responses were uniform, cytogenetic responses were variable. Complete cytogenetic responses occurred in 6 patients, with 4 also in remission by fluorescent in situ hybridization and/or reverse-transcription-polymerase chain reaction. Six of 19 had partial and 7 of 19 no cytogenetic response. Several patients acquired additional clonal cytogenetic abnormalities during therapy, a finding with significant implications for prognosis and laboratory monitoring in imatinib mesylate-treated CML patients. PMID- 12091334 TI - Low-dose total body irradiation followed by allogeneic lymphocyte infusion may induce remission in patients with refractory hematologic malignancy. AB - Allogeneic stem cell transplantation is curative for certain cancers, but the high doses of chemotherapy/radiotherapy lead to toxicity. Here, we treat patients with refractory cancer with 100 cGy total body irradiation (TBI) followed by infusion of nonmobilized pheresed allogeneic peripheral blood cells. Twenty-five patients, with a median age of 47 years, with refractory cancers were enrolled. Eighteen patients received sibling and 7 received unrelated cord blood cells. Donor chimerism was assessed at weeks 1, 2, 3, 4, and 8 after transplantation. Seven patients with solid tumors received a sibling transplant and 6 received a cord blood transplant; none achieved donor chimerism, but 1 treated at the higher dose level of 1 x 10(8) CD3+ cells/kg had a transient nodal response. Twelve patients with hematologic malignancies were treated; 1 received a cord blood transplant and 11 received sibling donor cells. Nine of these 11 patients achieved donor chimerism, ranging from 5% to 100%. Four patients had sustained complete remission of their cancers, including one patient with transient 5% donor chimerism. The development of chimerism correlated with hematologic malignancy (P <.001), total previous myelotoxic chemotherapy (P <.001), T-cell dose (P =.03), and graft-versus-host disease (P =.01). Tumor response correlated with donor chimerism (P =.01). Engraftment was achieved in patients with hematologic malignancies who had been heavily pretreated, suggesting the degree of immunosuppression may be a determinant of engraftment. Low-dose TBI and allogeneic lymphocyte infusion may induce remission in patients with refractory hematologic malignancy. PMID- 12091335 TI - Magnetic resonance imaging of fetal bone marrow for quantitative definition of the human fetal stem cell compartment. AB - Magnetic resonance imaging (MRI) can be used to distinguish bone marrow (BM) from cartilage and may therefore be used to measure BM volume in intact bones. We used MRI to measure the total human fetal BM volume in intact fetuses during the second trimester of pregnancy and determined the contribution of the individual bones to the total compartment. The total BM volume ranged from 934 microL at 17 to 18 weeks to 4563 microL at 22 to 23 weeks of gestation. The largest contributor to the total BM volume was the spine, constituting 26.4% +/- 2.7% of the total volume. By analyzing leukocyte content and percentages of CD34+ cells, lymphocytes, granulocytes, and monocytes of determined volumes, absolute numbers of these cell populations in BM could be measured. The cellular composition of the BM compartment did not significantly change throughout the second trimester of gestation. Absolute white blood cell counts per fetus increased from 111 x 10(6) at 16 to 17 weeks to 1229 x 10(6) at 21 to 22 weeks. The absolute numbers of CD34+ cells increased from 25 x 10(6) at 16 to 17 weeks to 256 x 10(6) at 21 to 22 weeks. Similar analysis of liver and spleen revealed comparable absolute numbers of CD34+ cells in BM and liver throughout the second trimester of gestation. In fetal liver, CD34+ cells differentiate into red cells, myeloid cells, and platelets, while lymphopoiesis mainly occurs in BM or spleen. Combining MRI and cell counts provides a method to quantify specific cell populations in fetal compartments. This study may enable better evaluation of fetal diagnostics and therapies. PMID- 12091336 TI - Runx1 is essential for hematopoietic commitment at the hemangioblast stage of development in vitro. AB - In this report we demonstrate a role for Runx1 (AML1) at the hemangioblast stage of hematopoietic and endothelial development in embryonic stem (ES) cell-derived embryoid bodies (EBs). Runx1 is expressed in EBs during the appearance of precursors with hemangioblast properties, the blast colony-forming cells (BL CFCs). Cell sorting studies revealed that all BL-CFCs within EBs express Runx1. Runx1-deficient EBs consistently generate 10- to 20-fold fewer blast colonies than wild-type controls and display a complete block in definitive hematopoiesis. Despite this defect, Runx1-/- EBs and yolk sacs from mutant embryos generate normal numbers of primitive erythroid precursors. These observations clearly demonstrate that Runx1 functions early in hematopoietic development, and they support the interpretation that the primitive erythroid lineage is established early by a subset of BL-CFCs that develop in a Runx1-independent fashion. PMID- 12091337 TI - Prostaglandin-E2 enhances EPO-mediated STAT5 transcriptional activity by serine phosphorylation of CREB. AB - Erythroid colony formation in response to erythropoietin (EPO) stimulation is enhanced by costimulating the cells with prostaglandin-E2 (PGE2). The present study further analyzed the underlying mechanisms and demonstrated that EPO mediated STAT5 transactivation in the erythroid AS-E2 cell line was enhanced 6 fold by PGE2 (10 microM), without affecting the STAT5 tyrosine phosphorylation or STAT5-DNA binding. Moreover, the PGE2-enhancing effect was independent of STAT5 serine phosphorylation. In AS-E2 cells STAT5 is constitutively phosphorylated on Ser780 (STAT5A) and EPO-dependently phosphorylated on Ser726/731 (STAT5A/STAT5B), but overexpression of STAT5 serine mutants did not affect STAT5 transactivation. In addition, PGE2 did not affect STAT5 serine phosphorylation. Instead, the stimulatory effect of PGE2 on STAT5 signaling could be mimicked by dibutyryl cyclic adenosine monophosphate (cAMP) and the phosphodiesterase inhibitor IBMX, suggesting that the effect was mediated by cAMP. Activation of the cAMP pathway resulted in cAMP-response element binding protein (CREB) phosphorylation, which was sustained in the presence of EPO plus PGE2 and transient on EPO stimulation alone. The costimulatory effect of PGE2 on EPO-mediated STAT5 transactivation was inhibited by overexpression of serine-dead CREB or protein kinase A (PKA) inhibitor (PKI), in contrast to EPO-mediated transactivation, which was PKA independent. Furthermore, CREB-binding protein (CBP)/p300 was shown to be involved in EPO-mediated STAT5 transactivation, and a CBP mutant with increased affinity for CREB resulted in an additional enhancement of the PGE2 effect. Finally, we demonstrated that the STAT5 target genes Bcl-X, SOCS2, and SOCS3 were up-regulated by costimulation with PGE2. In summary, these studies demonstrate that PGE2 enhancement of EPO-induced STAT5 transactivation is mediated by the cAMP/PKA/CREB pathway. PMID- 12091338 TI - Anemia of chronic disease in rheumatoid arthritis is associated with increased apoptosis of bone marrow erythroid cells: improvement following anti-tumor necrosis factor-alpha antibody therapy. AB - Circumstantial evidence has implicated tumor necrosis factor alpha (TNF-alpha) in the pathogenesis of anemia of chronic disease (ACD) in rheumatoid arthritis (RA). We investigated the role of TNF-alpha in erythropoiesis of patients with active RA (n = 40) and the effect of anti-TNF-alpha antibody administration (cA2). Patients with RA had lower numbers of CD34+/CD71+ and CD36-/glycophorin A+ (glycoA+) bone marrow (BM) cells and increased proportions of apoptotic cells within the CD34+/CD71+ and CD36+/glycoA+ cell compartments, compared to healthy controls (n = 24). Erythroid burst-forming units (BFU-Es) obtained by BM mononuclear or purified CD34+ cells were significantly lower in RA patients compared to controls. These abnormalities were more pronounced among patients with ACD. Increased TNF-alpha levels in patient long-term BM culture supernatants inversely correlated with BFU-Es and hemoglobin levels and positively with the percentage of apoptotic CD34+/CD71+ and CD36+/glycoA+ cells. Following cA2 therapy, a normalization was documented in the number of CD34+/CD71+ and CD36 /glycoA+ cells, the number of BFU-Es, and the proportion of apoptotic CD34+/CD71+ and CD36+/glycoA+ cells, which was associated with a significant increase in hemoglobin levels compared to baseline. Recovery from anemia was more prominent in patients with ACD. The exogenous addition of an anti-TNF-alpha antibody in the cultures increased BFU-E number in patients prior to cA2 treatment but not after treatment, further substantiating the inhibitory role of TNF-alpha on patients' erythropoiesis. We conclude that TNF-alpha-mediated apoptotic depletion of BM erythroid cells may account for ACD in RA and that cA2 administration may ameliorate ACD in these patients by down-regulating the apoptotic mechanisms involved in erythropoiesis. PMID- 12091339 TI - Granulocyte inducer C/EBPalpha inactivates the myeloid master regulator PU.1: possible role in lineage commitment decisions. AB - Several transcription factors have been implicated as playing a role in myelopoiesis. PU.1, an ets-family transcription factor, is required for the development of myeloid and lymphoid lineages, whereas the transcription factor CCAAT-enhancer binding protein family member C/EBPalpha is essential for granulocyte development. We present here the first evidence that C/EBPalpha blocks the function of PU.1. PU.1 and C/EBPalpha interact physically and colocalize in myeloid cells. As a consequence of this interaction, C/EBPalpha can inhibit the function of PU.1 to activate a minimal promoter containing only PU.1 DNA-binding sites. We further demonstrate that the leucine zipper in the DNA binding domain of C/EBPalpha interacts with the beta3/beta4 region in the DNA binding domain of PU.1 and as a result displaces the PU.1 coactivator c-Jun. Finally, C/EBPalpha blocks PU.1-induced dendritic cell development from CD34+ human cord blood cells. The functional blocking of PU.1 by C/EBPalpha could be the mechanism by which C/EBPalpha inhibits cell fates specified by PU.1 and directs cell development to the granulocyte lineage. PMID- 12091340 TI - Ectopic expression of TAL-1 protein in Ly-6E.1-htal-1 transgenic mice induces defects in B- and T-lymphoid differentiation. AB - The tal-1 gene encodes a basic helix-loop-helix (bHLH) transcription factor required for primitive and definitive hematopoiesis. Additionally, ectopic activation of the tal-1 gene during T lymphopoiesis occurs in numerous cases of human T-cell acute lymphoblastic leukemia. With the use of transgenic mice, we show that, in adult hematopoiesis, constitutive expression of TAL-1 protein causes disorders in the hematopoietic lineages that normally switch off tal-1 gene expression during their differentiation process. Myelopoiesis was characterized by a moderate increase of myeloid precursors and by Sca-1 antigen persistence. Although no lymphoid leukemia was observed, T lymphopoiesis and B lymphopoiesis were severely impaired. Transgenic mice showed reduced thymic cellularity together with a decrease in double-positive cells and a concurrent increase in the single-positive population. B cells exhibited a differentiation defect characterized by a reduction of the B-cell compartment most likely because of a differentiation block upstream of the intermediate pro-B progenitor. B cells escaping this defect developed normally, but transgenic splenocytes presented a defect in immunoglobulin class switch recombination. Altogether, these results enlighten the fine-tuning of TAL-1 expression during adult hematopoiesis and indicate why TAL-1 expression has to be switched off in the lymphoid lineages. PMID- 12091341 TI - Mutations associated with hemophilia A in the 558-565 loop of the factor VIIIa A2 subunit alter the catalytic activity of the factor Xase complex. AB - The 558-565 loop region in the A2 subunit of factor (F) VIIIa forms a direct interface with FIXa. We have expressed and purified B-domainless FVIII (FVIII(WT)) and B-domainless FVIII containing the hemophilia A-associated mutations Ser558Phe, Val559Ala, Asp560Ala, Gln565Arg, and the activated protein C cleavage site mutant Arg562Ala. Titration of FVIIIa in FXa generation assays showed that the mutant and wild-type proteins had similar functional affinities for FIXa (dissociation constant [K(d)] values approximately 5 nM-20 nM and approximately 100 nM-250 nM in the presence and absence of phospholipid, respectively). The catalytic activities of the factor Xase complex composed of the hemophilia A-associated FVIII species were markedly reduced both in the presence and absence of phospholipid. FVIII(WT) and FVIII(Arg562Ala) showed catalytic rate constant (k(cat)) values of approximately 60 minute(-1) in the presence of phospholipid, whereas the hemophilia A-associated mutants showed k(cat) values ranging from 3.3 minute(-1) to 7.5 minute(-1). In the absence of phospholipid, all k(cat) values were reduced but FVIII(WT) and FVIII(Arg562Ala) retained higher activities as compared with the hemophilic mutant FVIII forms. Fluorescence anisotropy experiments using fluorescein-modified FIXa confirmed that all FVIII forms interacted with FIXa. However, the presence of factor X yielded minimal increases in anisotropy observed with the mutant factor VIII forms, consistent with their reduced activity. These results show that residues within the 558-565 loop are critical in modulating FIXa enzymatic activity but do not contribute significantly to the affinity of FVIIIa for FIXa. PMID- 12091342 TI - A role for glycoprotein Ib in Streptococcus sanguis-induced platelet aggregation. AB - Numerous studies have implicated bacteria in cardiovascular disease, but there is a paucity of information on the mechanism involved. In this study we show how the common oral bacterium Streptococcus sanguis can directly interact with platelets, resulting in activation and aggregate formation. Platelet aggregation was dependent on glycoprotein IIb/IIIa (GPIIb/IIIa) and thromboxane. Platelets could also directly bind to S sanguis, but this interaction was not inhibited by GPIIb/IIIa antagonists. Antibodies to GPIb could inhibit both platelet aggregation and platelet adhesion to bacteria. This suggested a direct interaction between GPIb and S sanguis; however, this interaction did not require von Willebrand factor, the normal ligand for GPIb. By use of a range of monoclonal antibodies to GPIb and the enzyme mocharagin, which cleaves GPIb at amino acid 282, the interaction was localized to a region within the N-terminal 1 225 portion of GPIbalpha. Furthermore S sanguis failed to induce aggregation of platelets from a patient with Bernard-Soulier disease, the organism bound to Chinese hamster ovary cells transfected with the GPIbalpha gene but did not bind to mock-transfected cells and biotin-labeled S sanguis cells bound to purified GPIb in ligand blots. It is suggested that the interaction between S sanguis and GPIb is important in the pathogenesis of infective endocarditis and may also play a contributory role in some cases of myocardial infarction. PMID- 12091343 TI - C5a stimulates production of plasminogen activator inhibitor-1 in human mast cells and basophils. AB - We have recently shown that resting human mast cells (MCs) produce tissue-type plasminogen activator (t-PA) without simultaneously expressing plasminogen activator inhibitor 1 (PAI-1). In the present study we have identified the anaphylatoxin rhC5a as a potent inducer of PAI-1 expression in human MCs and basophils. In primary human skin MCs and primary blood basophils, exposure to rhC5a was followed by an increase from undetectable to significant levels of PAI 1. In addition, rhC5a induced a concentration- and time-dependent increase in PAI 1 antigen in the MC line HMC-1 and the basophil cell line KU-812 and increased the expression of PAI-1 mRNA in HMC-1. In conditioned media of HMC-1 treated with rhC5a, active PAI-1 could be detected. A simultaneous loss of t-PA activity in conditioned media from the same cells indicated that rhC5a-induced PAI-1 was capable of inhibiting the enzymatic activity of coproduced t-PA. Correspondingly, the levels of t-PA-PAI-1 complexes increased in rhC5a-treated cells. When HMC-1 cells were incubated with pertussis toxin or anti-C5a receptor antibodies, the effect of rhC5a on PAI-1 production was completely abolished. Treatment of C5a with plasmin resulted in loss of its ability to induce PAI-1 production in MCs. Considering the suggested role for MCs and components of the complement system in the development of cardiovascular diseases, we hypothesize that MCs, by producing t-PA in a resting state and by expressing PAI-1 when activated by C5a, might participate in the modulation of the balance between proteases and protease inhibitors regulating tissue injury and repair in such disease processes. PMID- 12091344 TI - Functional characterization of recombinant FV Hong Kong and FV Cambridge. AB - In factor V (FV) Cambridge (Arg306Thr) and Hong Kong (Arg306Gly), a cleavage site for anticoagulant activated protein C (APC), which is crucial for the inactivation of FVa, is lost. Although patients carrying FV Hong Kong have a normal APC response, those with FV Cambridge were reported to be APC resistant. To elucidate the molecular characteristics of the 2 FV mutants, we recreated them in a recombinant system and evaluated their functional properties. The 2 FV variants yielded identical APC resistance patterns, with APC responses being intermediate to those of wild-type FV and FV Leiden (Arg506Gln), which is known to be associated with the APC resistance phenotype. In the absence of protein S, APC mediated FVa inactivation curves obtained with the 2 variants were identical, resulting in partial FVa inactivation. In the presence of protein S, both FVa variants were almost completely inactivated because of protein S stimulation of the cleavage at Arg679. In a FVIIIa degradation system, both FV variants demonstrated slightly impaired APC cofactor activity. The ability of APC to cleave at Arg506 and at Arg679 in FVa Cambridge and Hong Kong and the slight decrease in APC cofactor activity of the 2 FV variants may explain the low thrombotic risk associated with these Arg306 mutations. In conclusion, we demonstrate that recombinant FV Cambridge and Hong Kong behave identically in in vitro assays and provide a mechanism for the low thrombotic risk associated with these FV mutations. PMID- 12091345 TI - The N-terminal peptide of PSGL-1 can mediate adhesion to trauma-activated endothelium via P-selectin in vivo. AB - P-selectin glycoprotein ligand-1 (PSGL-1) is present on leukocytes and is the major ligand for endothelial expressed P-selectin. A variety of studies strongly suggests that the N-terminal region of PSGL-1 contains the binding site for P selectin. We hypothesized that this relatively small N-terminal peptide of PSGL-1 is sufficient to support adhesion to P-selectin in vivo. To test this hypothesis, we coated 2 microm-diameter microspheres with a recombinant PSGL-1 construct, termed 19.ek.Fc. The 19.ek.Fc construct consists of the first 19 N-terminal amino acids of mature PSGL-1 linked to an enterokinase cleavage site that, in turn, is linked to human immunoglobulin G Fc. The 19.ek.Fc-coated microspheres were injected into the jugular vein of mice. Intravital microscopy of postcapillary venules within the cremaster muscle of mice revealed that a significantly greater number of 19.ek.Fc microspheres rolled compared with control microspheres. The number of rolling 19.ek.Fc microspheres was significantly diminished by pretreatment of the mice with a monoclonal antibody to P-selectin or by pretreatment of the 19.ek.Fc microspheres with a monoclonal antibody to PSGL-1. Combined, the results indicate that the N-terminal peptide of PSGL-1 can mediate adhesion to trauma-activated microvascular endothelium via P-selectin in vivo. PMID- 12091346 TI - Activated protein C cleaves factor Va more efficiently on endothelium than on platelet surfaces. AB - The protein C/protein S system is known to regulate thrombin generation in vivo by cleaving factors Va and VIIIa. We have examined the activity of activated protein C in several tissue factor-initiated models of coagulation. We used 4 models: monocytes as the tissue factor source with platelets as the thrombin generating surface; endothelial cells as the tissue factor source with platelets as the thrombin-generating surface; endothelial cells as both the tissue factor source and the thrombin-generating surface; and relipidated tissue factor with lipid vesicles providing the surface for thrombin generation. With the lipid surface, activated protein C dose-dependently reduced thrombin generation. Similarly, when endothelial cells provided the only surface for thrombin generation, activated protein C dose-dependently decreased thrombin generation significantly. By contrast, whenever platelets were present, activated protein C only minimally affected the amount of thrombin generated. When endothelial cells were the tissue factor source with platelets providing the surface for thrombin generation, activated protein C did increase the time until the burst of thrombin generation but had minimal effects on the total amount of thrombin generated. Activated protein C had essentially no effect on thrombin generation when monocytes were the tissue factor source with platelets providing the surface for thrombin generation. From the studies reported here, we conclude that in vivo, despite the important role of the protein C system in regulating thrombosis, activated protein C does not serve as a primary regulator of platelet-dependent thrombin generation. PMID- 12091347 TI - Generation of minor histocompatibility antigen HA-1-specific cytotoxic T cells restricted by nonself HLA molecules: a potential strategy to treat relapsed leukemia after HLA-mismatched stem cell transplantation. AB - Successful stem cell transplantation (SCT) across HLA barriers can be performed with cord blood, megadoses of stem cells, or with nonmyeloablative conditioning strategies. Because the HLA-mismatched transplants are often T-cell depleted, leukemia relapse rates are high. Treatment of relapsed leukemia after HLA mismatched SCT is difficult. A novel potential strategy to treat relapsed leukemia after HLA-mismatched SCT is the use of patients' mismatched HLA molecules as antigen-presenting molecules to generate hematopoietic system specific cytotoxic T cells (CTLs) from the stem cell donor. Adoptive transfer of these hematopoietic system-specific CTLs that are restricted by nonself HLA molecules may eliminate leukemia without affecting the patient's nonhematopoietic cells or donor hematopoietic cells. We investigated the feasibility of this strategy using the hematopoietic system-specific minor histocompatibility antigen HA-1, which is known to induce HLA-A2-restricted CTLs. HLA-A2(-) peripheral blood mononuclear cells were stimulated with HLA-A2(+) T2 cells pulsed with synthetic HA-1 peptide or with dendritic cells transduced with the HA-1 cDNA. Tetrameric HLA-A2/HA-1 peptide complexes were used to monitor and enrich HA-1-specific CTLs. In the alloreactive cultures, HA-1-specific CTLs were enriched up to 7% by 3 rounds of antigen-specific stimulations and up to 87% by fluorescence-activated cell sorting of tetramer-positive T cells. The HA-1-specific CTLs showed specific lysis of the relevant target cells, including leukemic cells. Because the polyclonal CTL cultures also contained natural killer cells and allo-HLA-A2 specific CTLs, CTL clones were generated that showed the expected HA-1 specificity only. Thus, HA-1-specific CTLs restricted by nonself HLA-A2 molecules can be generated in an HLA-A2-mismatched setting. PMID- 12091348 TI - Presence of chromogranin-derived antimicrobial peptides in plasma during coronary artery bypass surgery and evidence of an immune origin of these peptides. AB - Chromogranin A (CGA) and chromogranin B (CGB) are acidic proteins stored in secretory organelles of endocrine cells and neurons. In addition to their roles as helper proteins in the packaging of peptides, they may serve as prohormones to generate biologically active peptides such as vasostatin-1 and secretolytin. These molecules derived from CGA and CGB, respectively, possess antimicrobial properties. The present study demonstrates that plasmatic levels of both vasostatin-1 and secretolytin increase during surgery in patients undergoing cardiopulmonary bypass (CPB). Vasostatin-1 and secretolytin, initially present in plasma at low levels, are released just after skin incision. Consequently, they can be added to enkelytin, an antibacterial peptide derived from proenkephalin A, for the panoply of components acting as a first protective barrier against hypothetical invasion of pathogens, which may occur during surgery. CGA and CGB, more commonly viewed as markers for endocrine and neuronal cells, were also found to have an immune origin. RNA messengers coding for CGB were amplified by reverse transcription-polymerase chain reaction in human monocytes, and immunocytochemical analysis by confocal microscopy revealed the presence of CGA or CGB or both in monocytes and neutrophils. A combination of techniques including confocal microscopic analysis, mass spectrometry measurement, and antibacterial tests allowed for the identification of the positive role of interleukin 6 (IL-6) in the secretolytin release from monocytes in vitro. Because IL-6 release is known to be strongly enhanced during CPB, we suggest a possible relationship between IL-6 and the increased level of secretolytin in patients undergoing CPB. PMID- 12091349 TI - Induced disruption of the transforming growth factor beta type II receptor gene in mice causes a lethal inflammatory disorder that is transplantable. AB - Recent studies in mouse models deficient in transforming growth factor beta (TGF beta) signaling have documented TGF-beta as one of the major regulators of immune function. TGF-beta1-null animals demonstrated massive autoimmune inflammation affecting multiple organs, but attempts to transfer the phenotype to normal animals by bone marrow transplantation only resulted in minor inflammatory lesions. We wanted to ask whether a lethal inflammatory phenotype would develop following transplantation of bone marrow deficient for the TGF-beta type II receptor (TbetaRII) gene to normal recipient animals. The TbetaRII-null mutation would generate a cell autonomous phenotype that cannot be reverted by the influence of endocrine or paracrine TGF-beta derived from the recipient animal. We have generated conditional knockout mice in which the TbetaRII gene is disrupted upon induction with interferon-alphabeta or polyI:polyC. We show that induction of TbetaRII gene disruption in these mice by polyI:polyC results in a lethal inflammatory disease. Importantly, bone marrow from conditional knockout mice transferred to normal recipent mice caused a similar lethal inflammation, regardless of whether induction of TGF-beta receptor deficiency occurred in donor animals before, or in recipient animals after transplantation. These results show that TGF-beta signaling deficiency within cells of hematopoietic origin is sufficient to cause a lethal inflammatory disorder in mice. This animal model provides an important tool to further clarify the pathogenic mechanisms in animals deficient for TGF-beta signaling and the importance of TGF-beta to regulate immune functions. PMID- 12091350 TI - Identification of CD8alpha+CD11c- lineage phenotype-negative cells in the spleen as committed precursor of CD8alpha+ dendritic cells. AB - CD8alpha+ dendritic cells (DCs) represent a functionally distinct DC subset in vivo, which plays a critical role in initiating various cellular immune responses. However, the committed precursor of CD8alpha+ DCs remains to be identified. We reported here that murine splenic CD8alpha+CD11c- lineage phenotype (Lin)- cells could differentiate into CD8alpha+ DCs in vivo after intravenous transplantation. Immunohistochemistry staining showed that donor derived DCs mainly located in T-cell areas of the spleen. Functionally, these CD8alpha+CD11c-Lin- cell-derived DCs were capable of stimulating allogenic T-cell response, as well as secreting bioactive interleukin 12 p70 and interferon gamma. Freshly isolated CD8alpha+CD11c-Lin- cells expressed CC chemokine receptor (CCR)2, CCR5, and CCR7 messenger RNA, whereas CD8alpha+ DCs derived from CD8alpha+CD11c-Lin- cells further obtained the expression of CCR6 and macrophage derived chemokine. Flow cytometry analysis showed that CD8alpha+CD11c-Lin- cells were identified in bone marrow and lymph nodes. Moreover, transplanted splenic CD8alpha+CD11c-Lin- cells could also home to thymus and lymph nodes and were capable of developing into CD8alpha+ DCs in these locations. However, CD8alpha+CD11c-Li- cells failed to differentiate into CD8alpha- DCs, T cells, natural killer cells, or other myeloid lineage cells in irradiated chimeras. Taken together, all these findings suggest that CD8alpha+CD11c-Lin- cells are a committed precursor of CD8alpha+ DCs. PMID- 12091351 TI - Single-cell analysis of CD30+ cells in lymphomatoid papulosis demonstrates a common clonal T-cell origin. AB - Lymphomatoid papulosis (LyP) represents an intriguing cutaneous T-cell lymphoproliferative disorder with a histologic appearance resembling malignant lymphoma. This finding strongly contrasts with the benign clinical course of the disease. However, in 10% to 20% of cases, LyP can precede, coexist with, or follow malignant lymphoma. In these cases, the same T-cell population has been shown to be present in the LyP as well as in the associated lymphoma. In most LyP cases, there is-despite the sometimes extremely long course of the disease-no evolution of a secondary lymphoma. The investigation of these uncomplicated LyP cases for the presence of clonal T-cell receptor rearrangements has produced heterogeneous results. This might be explained by biologic or technical reasons arising from analyzing whole tissue DNA extracts. To definitively clarify whether the large atypical CD30(+) cells in LyP without associated lymphoma all belong to the same clone or represent individually rearranged T cells, we analyzed the T cell receptor-gamma rearrangements of single CD30+ as well as of single CD30- cells isolated from 14 LyP lesions of 11 patients. By using this approach we could demonstrate that the CD30+ cells represent members of a single T-cell clone in all LyP cases. Moreover, in 3 patients the same CD30+ cell clone was found in anatomically and temporally separate lesions. In contrast, with only a few exceptions, the CD30- cells were polyclonal in all instances and unrelated to the CD30+ cell clone. Our results demonstrate that LyP unequivocally represents a monoclonal T-cell disorder of CD30+ cells in all instances. PMID- 12091352 TI - Inhibition of constitutively active forms of mutant kit by multitargeted indolinone tyrosine kinase inhibitors. AB - Mutations in the proto-oncogene c-kit, including point mutations, deletions, or duplications in the negative regulatory juxtamembrane (JM) domain or point mutations in the catalytic domain, have been observed in human and canine cancers and often result in constitutive activation of Kit in the absence of ligand binding. To identify a receptor tyrosine kinase (RTK) inhibitor capable of blocking the function of mutant Kit, we evaluated 3 indolinones (SU11652, SU11654, and SU11655) that act as competitive inhibitors of adenosine triphosphate binding to several members of the split kinase family of RTKs, including VEGFR, FGFR, PDGFR, and Kit. Mast cell lines expressing either wild type (WT) Kit, a point mutation in the JM domain, a tandem duplication in the JM domain, or a point mutation in the catalytic domain were used for these studies. All 3 indolinones inhibited phosphorylation of WT Kit in the presence of stem cell factor at concentrations as low as 0.01 microM. Autophosphorylation of both JM mutants was inhibited at 0.01 to 0.1 microM, resulting in cell cycle arrest within 24 hours, whereas autophosphorylation of the catalytic domain mutant was inhibited at 0.25 to 0.5 microM, resulting in cell death within 24 hours. poly(ADP-ribose) polymerase (PARP) cleavage was noted in all Kit mutant lines after indolinone treatment. In summary, SU11652, SU11654, and SU11655 are effective RTK inhibitors capable of disrupting the function of all forms of mutant Kit. Because the concentrations of drug necessary for receptor inhibition are readily achievable and nontoxic in vivo, these compounds may be useful in the treatment of spontaneous cancers expressing Kit mutations. PMID- 12091353 TI - Strategy for the treatment of acute myelogenous leukemia based on folate receptor beta-targeted liposomal doxorubicin combined with receptor induction using all trans retinoic acid. AB - Up-regulation of folate receptor (FR) type-beta in acute myelogenous leukemia (AML) by all-trans retinoic acid (ATRA) and its restricted normal tissue distribution makes it a potential target for therapeutic intervention. The FR beta in peripheral blood granulocytes was unable to bind folate and appeared to have a variant GPI membrane anchor, evident from its insensitivity to phosphatidylinositol-specific phospholipase C but not nitrous acid. Granulocyte FR-beta lacked mutations, and neither deglycosylation nor detergent solubilization restored folate binding. The posttranslational modification causing its nonfunctionality was evidently absent in FR-beta from AML cells from patient marrow, which bound folate. From flow cytometric analysis of 78 AML bone marrow specimens of different subtypes, 68% expressed FR-beta, most of which were also CD34+. In model cell lines that are FR - (KG-1a, L1210, and Chinese hamster ovary [CHO]) or FR + (KG-1, L1210 JF, and recombinant CHO-FR-beta), selective FR mediated binding and cytotoxicity was obtained using folate-coated liposomes encapsulating fluorescent calcein (f-L-calcein) and doxorubicin (f-L-DOX), respectively, which could be blocked by 1 mM free folic acid. In the FR-beta expressing KG-1 human AML cells, treatment with ATRA further increased this specificity. In mouse ascites leukemia models generated using L1210JF or KG-1 cells, increased median survival times were obtained with f-L-DOX treatment compared to nontargeted L-DOX. In the KG-1 model, ATRA treatment increased the cure rate with f-L-DOX from 10% to 60%. The above combined data from our 2 laboratories further support the feasibility and potential usefulness of selective ATRA-facilitated liposomal drug delivery in FR-beta + AMLs. PMID- 12091354 TI - ATM mutations are rare in familial chronic lymphocytic leukemia. AB - It is now recognized that a subset of B-cell chronic lymphocytic leukemia (CLL) is familial. The genetic basis of familial CLL is poorly understood, but recently germ line mutations in the Ataxia Telangiectasia (ATM) gene have been proposed to confer susceptibility to CLL. The evidence for this notion is, however, not unequivocal. To examine this proposition further we have screened the ATM gene for mutations in CLLs from 61 individuals in 29 families. Truncating ATM mutations, including a known ATM mutation, were detected in 2 affected individuals, but the mutations did not cosegregate with CLL in the families. In addition, 3 novel ATM missense mutations were detected. Common ATM missense mutations were not overrepresented. The data support previous observations that ATM mutation is associated with B-CLL. However, ATM mutations do not account for familial clustering of the disease. PMID- 12091355 TI - Soluble syndecan-1 promotes growth of myeloma tumors in vivo. AB - Syndecan-1 (CD138) is a transmembrane heparan sulfate-bearing proteoglycan expressed by most myeloma plasma cells that regulates adhesion, migration, and growth factor activity. In patients with myeloma, shed syndecan-1 accumulates in the bone marrow, and high levels of syndecan-1 in the serum are an indicator of poor prognosis. To test the effect of soluble syndecan-1 on tumor cell growth and dissemination, ARH-77 B-lymphoid cells were engineered to produce a soluble form of syndecan-1. Controls included vector only (neo)-transfected cells and cells transfected with full-length syndecan-1 complementary DNA that codes for the cell surface form of syndecan-1. Assays reveal that all 3 transfectants have similar growth rates in vitro, but cells expressing soluble syndecan-1 are hyperinvasive in collagen gels relative to controls. When injected into the marrow of human bones that were implanted in severe combined immunodeficient mice, tumors formed by cells expressing soluble syndecan-1 grow faster than tumors formed by neo transfected cells or by cells expressing cell surface syndecan-1. In addition, cells bearing cell surface syndecan-1 exhibit a diminished capacity to establish tumors within the mice as compared with both neo- and soluble syndecan-1 transfected cells. Tumor cell dissemination to a contralateral human bone is detected significantly more often in the tumors producing soluble syndecan-1 than in controls. Thus, high levels of soluble syndecan-1 present in patients with myeloma may contribute directly to the growth and dissemination of the malignant cells and thus to poor prognosis. PMID- 12091356 TI - Cytogenetic profile of childhood and adult megakaryoblastic leukemia (M7): a study of the Groupe Francais de Cytogenetique Hematologique (GFCH). AB - To draw the cytogenetic profile of childhood and adult acute megakaryoblastic leukemia (M7), the Groupe Francais de Cytogenetique Hematologique collected 53 cases of M7 (30 children and 23 adults). Compared to other acute myeloid leukemias, M7 is characterized by a higher incidence of abnormalities, a higher complexity of karyotypes, and a different distribution of abnormalities among children and adults. Nine cytogenetic groups were identified: normal karyotypes (group 1), patients with Down syndrome (group 2), numerical abnormalities only (group 3), t(1;22)(p13;q13) or OTT-MAL transcript (group 4), t(9;22)(q34;q11) (group 5), 3q21q26 (group 6), -5/del(5q) or -7/del(7q) or both (group 7), i(12)(p10) (group 8), and other structural changes (group 9). Groups 1, 2, 3, and 4 were exclusively composed of children (except one adult in group 3), whereas groups 5, 6, 7, and 8 were mainly made up of adults. The main clinical and hematologic features of these groups were described. No new recurrent abnormality was identified, but mapping of all breakpoints allowed us to specify several possible hot spots of rearrangement: 17q22-23, 11q14-21, 21q21-22, and 16q21-22 23. Although 90.5% of cases had no documented antecedent hematologic disorder or exposure to chemotherapy or radiotherapy, the morphologic and the cytogenetic findings indicated that M7 might be a secondary leukemia more often than suggested by preceding history, particularly among adults. The concurrent analyses of morphologic and cytogenetic data also led us to assume that the initial precursor involved might be more immature in adult than in childhood M7. PMID- 12091357 TI - Targeting CB2 cannabinoid receptors as a novel therapy to treat malignant lymphoblastic disease. AB - In the current study, we examined whether ligation of CB2 receptors would lead to induction of apoptosis in tumors of immune origin and whether CB2 agonist could be used to treat such cancers. Exposure of murine tumors EL-4, LSA, and P815 to delta-9-tetrahydrocannabinol (THC) in vitro led to a significant reduction in cell viability and an increase in apoptosis. Exposure of EL-4 tumor cells to the synthetic cannabinoid HU-210 and the endogenous cannabinoid anandamide led to significant induction of apoptosis, whereas exposure to WIN55212 was not effective. Treatment of EL-4 tumor-bearing mice with THC in vivo led to a significant reduction in tumor load, increase in tumor-cell apoptosis, and increase in survival of tumor-bearing mice. Examination of a number of human leukemia and lymphoma cell lines, including Jurkat, Molt-4, and Sup-T1, revealed that they expressed CB2 receptors but not CB1. These human tumor cells were also susceptible to apoptosis induced by THC, HU-210, anandamide, and the CB2 selective agonist JWH-015. This effect was mediated at least in part through the CB2 receptors because pretreatment with the CB2 antagonist SR144528 partially reversed the THC-induced apoptosis. Culture of primary acute lymphoblastic leukemia cells with THC in vitro reduced cell viability and induced apoptosis. Together, the current data demonstrate that CB2 cannabinoid receptors expressed on malignancies of the immune system may serve as potential targets for the induction of apoptosis. Also, because CB2 agonists lack psychotropic effects, they may serve as novel anticancer agents to selectively target and kill tumors of immune origin. PMID- 12091358 TI - Monoclonal B lymphocytes with the characteristics of "indolent" chronic lymphocytic leukemia are present in 3.5% of adults with normal blood counts. AB - Molecular and cellular markers associated with malignant disease are frequently identified in healthy individuals. The relationship between these markers and clinical disease is not clear, except where a neoplastic cell population can be identified as in myeloma/monoclonal gammopathies of undetermined significance (MGUS). We have used the distinctive phenotype of chronic lymphocytic leukemia (CLL) cells to determine whether low levels of these cells can be identified in individuals with normal complete blood counts. CLL cells were identified by 4 color flow cytometric analysis of CD19/CD5/CD79b/CD20 expression in 910 outpatients over 40 years old. These outpatients were age- and sex-matched to the general population with normal hematologic parameters and no evident history of malignant disease. CLL phenotype cells were detectable in 3.5% of individuals at low level (median, 0.013; range, 0.002- 1.458 x 10(9) cells/L), and represented a minority of B lymphocytes (median, 11%; range, 3%-95%). Monoclonality was demonstrated by immunoglobulin light-chain restriction in all cases with CLL phenotype cells present and confirmed in a subset of cases by consensus-primer IgH-polymerase chain reaction. As in clinical disease, CLL phenotype cells were detected with a higher frequency in men (male-to-female ratio, 1.9:1) and elderly individuals (2.1% of 40- to 59-year-olds versus 5.0% of 60- to 89-year-olds, P =.01). The neoplastic cells were identical to good-prognosis CLL, being CD5+23+20(wk)79b(wk)11a(-)22(wk)sIg(wk)CD38-, and where assessed had a high degree (4.8%-6.6%) of IgH somatic hypermutation. The monoclonal CLL phenotype cells present in otherwise healthy individuals may represent a very early stage of indolent CLL and should be useful in elucidating the mechanisms of leukemogenesis. PMID- 12091359 TI - Immature CD34+CD19- progenitor/stem cells in TEL/AML1-positive acute lymphoblastic leukemia are genetically and functionally normal. AB - One important question in stem cell biology of childhood acute lymphoblastic leukemia (ALL) is whether immature CD34+CD19- cells are part of the leukemic cell clone. CD34+CD19- cells from the bone marrow of 9 children with TEL/AML1-positive ALL were purified by flow sorting and subjected to reverse transcriptase polymerase chain reaction (RT-PCR), fluorescence in situ hybridization, and methylcellulose cultures. In 3 of 8 patients analyzed by RT-PCR, no TEL/AML1 positive cells could be detected in the CD34+CD19- cell fraction. Altogether, the percentage of TEL/AML1-positive cells was low: 1.6% (n = 8; SD 2.2%) by nested real-time RT-PCR and 2.5% (n = 5; SD 2.6%) by fluorescence in situ hybridization. This correlated with the percentage of contaminating CD19+ leukemic cells in the CD34+CD19- cell fraction in 6 control sorts (mean 4.6%, SD 3.6%), indicating that the low levels of leukemic cells detected in the CD34+CD19- cell fraction could be attributed to sorter errors. Methylcellulose cultures in 3 patients provided further evidence that CD34+CD19- cells represent a candidate normal cell population. The clonogenicity of the CD34+CD19- cell fraction was similar to normal progenitors, including growth of primitive granulocyte, erythroid, macrophage, megakaryocyte colony-forming units. Each of 92 colonies from cultures with CD34+CD19- cells tested negative for TEL/AML1. In conclusion, our data support the hypothesis that the leukemia in TEL/AML1-positive childhood ALL originates in a CD19+ lymphoid progenitor. This has many therapeutic implications, eg, for purging of autologous stem cell products, flow cytometric monitoring of minimal residual disease, and targeting immunotherapy against the leukemic cell clone. PMID- 12091360 TI - Response of hairy cells to IFN-alpha involves induction of apoptosis through autocrine TNF-alpha and protection by adhesion. AB - Although hairy cell leukemia is uniquely sensitive to interferon-alpha (IFN alpha), the biologic basis for this phenomenon remains unclear. Here we examine the effects of IFN-alpha on cultured hairy cells (HCs), taking into account the possible modifying influence of cell adhesion. We make the novel observation that therapeutic concentrations of IFN-alpha kill nonadherent HCs by inducing apoptosis. In keeping with the persistence of HCs in tissues during therapy, such killing was inhibited by integrin-mediated adhesion to vitronectin or fibronectin. Exposure of HCs to IFN-alpha resulted in a marked increase in tumor necrosis factor-alpha (TNF-alpha) secretion. Furthermore, blocking antibodies to TNF-RI or TNF-RII protected HCs from IFN-alpha-induced apoptosis, demonstrating that such killing was mediated by TNF-alpha. In the absence of IFN-alpha, exogenous TNF-alpha did not induce HC apoptosis, showing that IFN-alpha sensitized HCs to the proapoptotic effect of autocrine TNF-alpha. This sensitization to TNF-alpha-induced killing was attributable to suppression of IAP (inhibitors of apoptosis) production known to be regulated by the cytoprotective nuclear factor-kappaB-dependent arm of TNF-alpha signaling. Moreover, engagement of the receptors for fibronectin or vitronectin prevented this IFN-alpha-induced down-regulation of IAPs. Understanding of the signals involved in the combined effects of IFN-alpha and TNF-alpha and abrogation of those induced by integrin engagement offers the possibility of sensitizing other malignant cells to IFN alpha-induced killing and thereby extending the therapeutic use of this cytokine. PMID- 12091361 TI - An animal model for human cellular immunotherapy: specific eradication of human acute lymphoblastic leukemia by cytotoxic T lymphocytes in NOD/scid mice. AB - Adoptive immunotherapy using in vitro-generated donor-derived cytotoxic T lymphocytes (CTLs) can be effective in the treatment of relapsed leukemia after allogeneic transplantation. To determine effector cell characteristics that result in optimal in vivo antileukemic efficacy, we developed an animal model for human CTL therapy. Nonobese diabetic/severe combined immunodeficiency (NOD/scid) mice were inoculated with either of 2 primary human acute lymphoblastic leukemia (ALL), denoted as SK and OF. Anti-SK and anti-OF CTLs were generated in vitro by repeated stimulation of donor peripheral blood mononuclear cells with either SK or OF cells. Both CTL lines displayed HLA-restricted reactivity against the original targets and non-major histocompatibility class (MHC)-restricted cross reactivity in vitro. The CTLs were administered intravenously weekly for 3 consecutive weeks to mice engrafted with either SK or OF leukemia. In 3 of 8 SK engrafted and anti-SK-treated mice, complete remissions were achieved in blood, spleen, and bone marrow. In the remaining 5 animals partial remissions were observed. In 4 of 4 OF-engrafted anti-OF-treated mice partial remissions were observed. The antileukemic effect of specific CTLs was exerted immediately after administration and correlated with the degree of HLA disparity of the donor patient combination. In cross-combination-treated animals, no effect on leukemic progression was observed indicating that in vivo antileukemic reactivity is mediated by MHC-restricted effector cells. The CTLs, however, displayed an impaired in vivo proliferative capacity. Ex vivo analysis showed decreased reactivity as compared to the moment of infusion. We therefore conclude that the model can be used to explore the requirements for optimal in vivo efficacy of in vitro- generated CTLs. PMID- 12091362 TI - Evidence for the involvement of a hematopoietic progenitor cell in systemic mastocytosis from single-cell analysis of mutations in the c-kit gene. AB - Mast cells are derived from multipotential hematopoietic progenitors and are clonally increased in systemic mastocytosis, a disease associated with point mutations of codon 816 (most commonly Asp816Val) of c-kit. To study the lineage relationship and the extent of expansion of cells derived from the mutated clone, we examined the occurrence of the Asp816Val c-kit mutation in genomic DNA of individual sorted peripheral blood T cells, B cells, and monocytes in patients with indolent systemic mastocytosis. The mutation was detected in varying frequencies in the genomic DNA of individual B cells and monocytes and bone marrow mast cells in patients with extensive disease. In B cells, the immunoglobulin repertoire was polyclonal, indicating that the mutation occurred before V(H)/(D)/J(H) recombination. These results show that mastocytosis is a disorder of a pluripotential hematopoietic progenitor cell that gives rise to B cells and monocytes in addition to mast cells and that the affected clone shows variable expansion in these lineages in the peripheral blood of patients with systemic mastocytosis. PMID- 12091363 TI - Novel "hybrid" iron chelators derived from aroylhydrazones and thiosemicarbazones demonstrate selective antiproliferative activity against tumor cells. AB - We previously demonstrated that 2-hydroxy-1-naphthylaldehyde isonicotinoyl hydrazone (311) and other aroylhydrazone chelators possess potent antineoplastic activity because of their ability to bind iron (Fe). From these studies, we identified structural components of the hydrazones that provide antineoplastic activity, namely the salicylaldehyde and 2-hydroxy-1-naphthylaldehyde moieties. A related group of chelators known as the thiosemicarbazones also show pronounced antitumor activity because of their ability to inhibit ribonucleotide reductase. Considering this, we designed a new series of "hybrid ligands" by condensation of the aldehydes described above with a range of thiosemicarbazides. The parent compound of these ligands is 2-hydroxy-1-naphthylaldehyde thiosemicarbazone (NT). Of 8 NT analogues, 3 chelators, namely NT, N4mT (2-hydroxy-1-naphthylaldehyde-4 methyl-3-thiosemicarbazone), and N44mT (2-hydroxy-1-naphthylaldehyde-4,4-dimethyl 3-thiosemicarbazone), showed high antiproliferative activity against SK-N-MC neuroepithelioma cells (50% inhibitory concentration [IC(50)] = 0.5-1.5 microM). Indeed, their activity was significantly (P <.0001) greater than that of desferrioxamine (DFO) (IC(50) = 22 microM). We demonstrate that 311, a 311 analogue (311m), and several NT-series chelators have significantly (P <.001) greater antiproliferative activity against tumor cells than against a range of normal cell types. For example, the IC(50) values of NT and N4mT in SK-N-MC neuroepithelioma cells were 0.5 microM, whereas for fibroblasts the IC(50) values were greater than 25 microM. Further, the effect of one of the most potent chelators (311m) on preventing the growth of bone marrow stem cell cultures was far less than that of doxorubicin and similar to that of cisplatin. These studies support the further development of these chelators as antiproliferative agents. PMID- 12091364 TI - Persistence of HTLV-I in blood components after leukocyte depletion. AB - The human T-cell leukemia virus HTLV-I is a transfusion-transmissible retrovirus targeting T lymphocytes for which screening is not currently undertaken in United Kingdom blood donors. The introduction of universal leukocyte depletion (LD) of the United Kingdom blood supply raises the question as to the degree of protection afforded by this procedure against HTLV-I transmission by blood components. HTLV-I viral DNA removal by leukocyte-depleting filters was assessed in units of whole blood and platelets by real-time quantitative polymerase chain reaction (PCR) and by nested PCR for HTLV-I Tax DNA. We examined HTLV-I removal by LD filters using a model system of blood units containing exogenous spiked HTLV-I-positive MT-2 cells at a relevant concentration and whole blood donations from asymptomatic HTLV-I carriers. T-lymphocyte removal was assessed in parallel by measurement of endogenous subset-specific CD3 mRNA. In the MT-2 model system we observed 3.5 log(10) to 4 log(10) removal of HTLV-I Tax DNA by filtration of whole blood and 2 log(10) to 3 log(10) removal across platelet filters with 13 of 16 whole blood and 8 of 8 platelet units still positive after filtration. Despite 3 log(10) to 4 log(10) viral removal, HTLV-I Tax DNA could be detected after whole blood filtration in asymptomatic carriers with viral loads above 10(8) proviral DNA copies/L. T-lymphocyte removal was also between 3.5 log(10) and 4.5 log(10). HTLV-I provirus removal was incomplete in the model system and in asymptomatic carriers with viral loads greater than 10(8) copies/L. These results suggest that LD alone may not provide complete protection from HTLV-I transmission by transfusion. PMID- 12091365 TI - Keratinocyte growth factor preserves normal thymopoiesis and thymic microenvironment during experimental graft-versus-host disease. AB - Thymus-dependent reconstitution of the peripheral T-cell compartment is critical for the successful outcome of bone marrow transplantation. However, graft-versus host disease (GVHD) affects thymic stromal function and thus prevents normal T cell maturation and selection. To determine whether cytoprotection of thymic epithelial cells (TECs) by keratinocyte growth factor (KGF) averts GVHD-related injury to the thymus, a nonirradiated murine parent-->F(1) transplantation model was investigated. Administration of KGF between days -3 and +3 of GVHD induction preserved normal thymic size, cellularity, and thymocyte phenotype when measured 2 weeks after transplantation and compared with saline-treated parent-->F(1) mice that received allogeneic transplants. Moreover, the characteristic GVHD-induced impairment in cell cycle progression of pro- and pre-T cells was prevented by KGF. However, the normal phenotypic and functional status of the thymus did not correlate with the higher number of GVHD-inducing mature donor T cells in thymi of KGF-treated mice. Importantly, extensive analysis of the different TEC populations within the thymic cortex and medulla revealed an almost normal stromal architecture and composition in GVHD mice treated with KGF. These observations are likely to reflect an indirect effect of KGF on thymopoiesis as KGF-receptor expression was demonstrated to be restricted to TECs. Thus, pharmacologic doses of KGF appear to exert a potent effect on TEC function, which in turn allows for normal T lymphopoiesis to occur during acute GVHD. PMID- 12091366 TI - Novel mutation in ferroportin1 is associated with autosomal dominant hemochromatosis. AB - Hemochromatosis is a common disorder characterized by excess iron absorption and accumulation of iron in tissues. Usually hemochromatosis is inherited in an autosomal recessive pattern and is caused by mutations in the HFE gene. Less common non-HFE-related forms of hemochromatosis have been reported and are caused by mutations in the transferrin receptor 2 gene and in a gene localized to chromosome 1q. Autosomal dominant forms of hemochromatosis have also been described. Recently, 2 mutations in the ferroportin1 gene, which encodes the iron transport protein ferroportin1, have been implicated in families with autosomal dominant hemochromatosis from the Netherlands and Italy. We report the finding of a novel mutation (V162del) in ferroportin1 in an Australian family with autosomal dominant hemochromatosis. We propose that this mutation disrupts the function of the ferroportin1 protein, leading to impaired iron homeostasis and iron overload. PMID- 12091367 TI - Autosomal dominant reticuloendothelial iron overload associated with a 3-base pair deletion in the ferroportin 1 gene (SLC11A3). AB - We describe a family with autosomal dominant inheritance of increased body iron stores characterized by raised serum ferritin concentration and normal transferrin saturation. Liver biopsy showed iron deposition in Kupffer cells without fibrosis. The clinical features of HFE-related hemochromatosis were absent, as were the Cys282Tyr and His63Asp mutations. Venesection therapy was poorly tolerated, suggesting a defect in iron release from reticuloendothelial stores. A 3-base pair deletion in exon 5 of the ferroportin 1 gene (SLC11A3) predicting Val162 deletion was found in affected members, but not in unaffected individuals or in 100 control subjects. Consensus structural predictions of the transmembrane helices showed that the deletion is in the extracellular loop between the third and fourth predicted transmembrane helices and lies within a spatial cluster of other known ferroportin 1 mutations. These results indicate that this extracellular cluster is functionally important for iron transport, and its disruption leads to iron overload. PMID- 12091368 TI - Absence of biologically important Kaposi sarcoma-associated herpesvirus gene products and virus-specific cellular immune responses in multiple myeloma. AB - Kaposi sarcoma-associated herpesvirus (KSHV) has been associated with several diseases, but the association between KSHV and multiple myeloma (MM) remains controversial. To address this issue, we studied patients with MM for the presence of viral RNA transcripts as well as KSHV-specific cellular immune responses. Highly sensitive reverse transcription-polymerase chain reaction assays for detection of viral transcripts of KSHV open reading frame (ORF) 26, ORF72, and ORF74 did not detect viral gene transcripts in long-term cultures of bone marrow stromal cells from 23 patients with MM. Moreover, sensitive assays for KSHV ORF65-specific and ORF73-specific cytotoxic T-lymphocyte (CTL) activity that readily and routinely detect CTLs specific for ORF65 and ORF73 in patients positive for human immunodeficiency virus and KSHV did not show any specific responses in 16 patients with MM, despite the presence of positive Epstein-Barr virus-specific CTLs in all cases. These data therefore do not show a biologically important association between ongoing KSHV infection and MM. PMID- 12091369 TI - MUTZ-3, a human cell line model for the cytokine-induced differentiation of dendritic cells from CD34+ precursors. AB - Many human myeloid leukemia-derived cell lines possess the ability to acquire a dendritic cell (DC) phenotype. However, cytokine responsiveness is generally poor, requiring direct manipulation of intracellular signaling mechanisms for differentiation. In contrast, the CD34+ human acute myeloid leukemia cell line MUTZ-3 responds to granulocyte macrophage- colony-stimulating factor (GM-CSF), interleukin 4 (IL-4), and tumor necrosis factor alpha (TNFalpha), cytokines known to be pivotal both in vivo and in vitro for DC generation from monocytes and CD34+ stem cells. In all respects, MUTZ-3 cells behave as the immortalized equivalent of CD34+ DC precursors. Upon stimulation with specific cytokine cocktails, they acquire a phenotype consistent with either interstitial- or Langerhans-like DCs and upon maturation (mDC), express CD83. MUTZ-3 DC display the full range of functional antigen processing and presentation pathways. These findings demonstrate the unique suitability of MUTZ-3 cells as an unlimited source of CD34+ DC progenitors for the study of cytokine-induced DC differentiation. PMID- 12091370 TI - High-dose cyclophosphamide for refractory autoimmune hemolytic anemia. AB - High-dose cyclophosphamide, without stem cell rescue, has been used successfully to treat aplastic anemia and other autoimmune disorders. To determine the safety and efficacy of high-dose cyclophosphamide among patients with severe refractory autoimmune hemolytic anemia, we treated 9 patients with cyclophosphamide (50 mg. kg(-1). d(-1) for 4 days) who had failed a median of 3 (range, 1-7) other treatments. The median hemoglobin before treatment was 6.7 g/dL (range, 5-10 g/dL). The median time to reach an absolute neutrophil count of 500/microL or greater was 16 days (range, 12-18 days). Six patients achieved complete remission (normal untransfused hemoglobin for age and sex), and none have relapsed after a median follow-up of 15 months (range, 4-29 months). Three patients achieved and continue in partial remission (hemoglobin at least 10 g/dL without transfusion support). High-dose cyclophosphamide was well tolerated and induced durable remissions in patients with severe refractory autoimmune hemolytic anemia. PMID- 12091371 TI - Paternal mosaicism proves the pathogenic nature of mutations in neutrophil elastase in severe congenital neutropenia. AB - Heterozygous mutations in neutrophil elastase have been detected in many sporadic cases of congenital neutropenia. However, a convincing pathogenetic mechanism has not been established, and it is unclear whether the effects of the mutant enzyme occur within the cell of production or are paracrine in nature. The healthy father of a patient was demonstrated to be mosaic for his daughter's Cys42Arg elastase mutation. Using semiquantitative polymerase chain reaction, approximately half of his T cells were shown to carry the mutation in contrast to less than 10% of neutrophils. Individual hematopoietic colonies grown from peripheral blood were heterozygous for the mutation or were homozygous wild type. These results demonstrate that precursors containing the mutation are selectively lost during myelopoiesis or fail to develop into neutrophils. This is the first in vivo confirmation of the pathogenic nature of elastase mutations in humans. The normal neutrophil count in the father suggests that the mutant elastase does not have paracrine effects. PMID- 12091372 TI - Von Willebrand factor-cleaving protease (ADAMTS13) in thrombocytopenic disorders: a severely deficient activity is specific for thrombotic thrombocytopenic purpura. AB - A severe deficiency in von Willebrand factor-cleaving protease (ADAMTS13) activity (< 5% that in normal plasma) has been observed in most patients with a diagnosis of thrombotic thrombocytopenic purpura (TTP) but not in those with a diagnosis of hemolytic uremic syndrome. However, ADAMTS13 deficiency has been claimed not to be specific for TTP, since it was observed in various thrombocytopenic and other conditions. We studied 68 patients with thrombocytopenia due to severe sepsis or septic shock (n = 17), heparin-induced thrombocytopenia (n = 16), idiopathic thrombocytopenic purpura (n = 10), or other hematologic (n = 15) or miscellaneous conditions (n = 10). Twelve of the 68 patients had subnormal levels of ADAMTS13 activity ( 80%). In contrast, c-mpl expression was heterogeneous in 60 patients with ET, 18 of whom (30%) presented with thrombosis at diagnosis, a significant difference from patients with a uniform c-mpl pattern (2 of 28; 7%; P =.026). In particular, the overrepresentation of thrombotic complications in patients with a heterogeneous c-mpl expression pattern was found mainly among patients with a significant percentage (10% to 40%) of weakly stained or c-mpl-negative megakaryocytes (heterogeneous-weak pattern; 13 of 30; 43%; P =.002). Accordingly, this pattern was associated with a 6.1-fold increased risk of thrombosis compared with that of patients with a uniform c-mpl pattern. In conclusion, the presence of a heterogeneous pattern of c-mpl distribution in bone marrow megakaryocytes could be a useful diagnostic criterion in the differential diagnosis of thrombocytosis. Furthermore, detection of a significant percentage of weakly stained or c-mpl-negative megakaryocytes can identify patients with a higher risk of thrombosis. PMID- 12091374 TI - Transcobalamin II 775G>C polymorphism and indices of vitamin B12 status in healthy older adults. AB - A common polymorphism (775G>C) in the vitamin B12 transport protein, transcobalamin II (TCII), has been identified in which proline replaces arginine at codon 259. We determined the influence of TCII genotype on indices of B12 status, including total serum B12, the amount of B12 bound to TCII (holoTCII), methylmalonic acid, and homocysteine, in 128 healthy older adults (ages 40-88 years). Mean total B12 and homocysteine concentrations were not significantly different among the 3 genotypes. Mean holoTCII concentration was significantly higher in those subjects homozygous for the proline form of TCII (PP) compared with those homozygous for the arginine form (RR) and heterozygotes (PR) (P 42 years) (P =.007). This study shows, for the first time in T cell-depleted transplantations, a positive correlation between the number of CD34+ cells and aGVHD and, also, that the number of CD3+ cells necessary to initiate aGVHD is lower than previously reported. PMID- 12091377 TI - Effects of pegylated recombinant human megakaryocyte growth and development factor in patients with idiopathic thrombocytopenic purpura. AB - We conducted a phase 1-2 clinical trial to evaluate the effect of pegylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF) in patients with chronic idiopathic thrombocytopenic purpura (ITP) refractory to standard therapy who had platelet counts below 30 x 10(9)/L. Four patients received PEG-rHuMGDF (0.5 microg/kg of body weight per day) by daily intravenous injection for up to 7 days. Administration of PEG-rHuMGDF increased platelet counts in 3 patients. A striking thrombocytosis occurred in 2 patients, whose platelet counts were elevated to more than 700 x 10(9)/L a week after the last administration of PEG-rHuMGDF and returned to baseline levels within 4 to 6 weeks. Before the platelet peak, the percentage of reticulated platelets increased transiently in 3 patients tested, including one patient who had no response. Bleeding episodes decreased after the start of PEG-rHuMGDF therapy. These results suggest that PEG-rHuMGDF might have a clinical benefit in ameliorating thrombocytopenia associated with ITP. PMID- 12091378 TI - Endogenous methylarginines regulate neuronal nitric-oxide synthase and prevent excitotoxic injury. AB - Nitric oxide (NO) has a critical role in neuronal function; however, high levels lead to cellular injury. While guanidino-methylated arginines (MA) including asymmetric dimethylarginine (ADMA) and N(G)-methyl-l-arginine (NMA) are potent competitive inhibitors of nitric oxide synthase (NOS) and are released upon protein degradation, it is unknown whether their intracellular concentrations are sufficient to critically regulate neuronal NO production and secondary cellular function or injury. Therefore, we determine the intrinsic neuronal MA concentrations and their effects on neuronal NOS function and excitotoxic injury. Kinetic studies demonstrated that the K(m) for l-arginine is 2.38 microm with a V(max) of 0.229 micromol mg(-1) min(-1), while K(i) values of 0.67 microm and 0.50 microm were determined for ADMA and NMA, respectively. Normal neuronal concentrations of all NOS-inhibiting MA were determined to be approximately 15 microm, while l-arginine concentration is approximately 90 microm. These MA levels result in >50% inhibition of NO generation from neuronal NOS. Down modulation or up-modulation of these neuronal MA levels, respectively, dramatically enhanced or suppressed NO-mediated excitotoxic injury. Thus, neuronal MA profoundly modulate NOS function and suppress NO mediated injury. Pharmacological modulation of the levels of these intrinsic NOS inhibitors offers a novel approach to modulate neuronal function and injury. PMID- 12091379 TI - Hypoxia and nitric oxide treatment confer tolerance to glucose starvation in a 5' AMP-activated protein kinase-dependent manner. AB - Hypoxia is a critical event for higher organisms, and cells and tissues react by increasing the oxygen supply by vasodilatation, angiogenesis, and erythropoiesis and maintaining cellular energy by increasing glycolysis and inhibiting anabolic pathways. Stimulation of glycolysis has been regarded as the main response that increases energy production during hypoxia; however, there is an obvious conflict during ischemia, because both the oxygen and glucose supply are insufficient. In this study, we found that exposure of HepG2 cells and normal fibroblasts to hypoxia induces cellular tolerance to glucose starvation. The tolerance induced by hypoxia is dependent on several amino acids, indicating a switch from glucose to amino acids as the energy source. When antisense RNA expression vector for 5' AMP-activated protein kinase or protein kinase B/Akt was transfected into HepG2 cells, the induction of tolerance to glucose was greatly inhibited, indicating that the tolerance was dependent on 5'-AMP-activated protein kinase and protein kinase B/Akt. Similar tolerance was induced by nitric oxide exposure. The tolerance induced was observed in various cells and may represent a previously unknown physiological response related to hypoxia-preconditioning and tumor progression:austerity. PMID- 12091380 TI - The role of ADAM 15 in glomerular mesangial cell migration. AB - Mesangial cells (MC) occupy the core of the renal glomerulus and are surrounded by a mesangial matrix. In certain diseases, MC migrate through this matrix into the pericapillary space. The mechanisms involved, however, are poorly understood. Members of the ADAM (A Disintegrin And Metalloproteinase) family of membrane proteins have the potential to be key modulators of cell-matrix interactions through the activities of their constituent domains. We have studied the possible role of ADAM 15 in human (H) MC migration in vitro. HMC ADAM 15 was expressed at low levels in serum-free medium but was increased during migration. Antibodies to the individual domains of ADAM 15 and the incorporation of antisense ADAM 15, (but not control oligonucleotide) inhibited this migration. Furthermore, inhibition of migration by the broad spectrum metalloproteinase inhibitor BB3103, demonstrated that metalloproteinase activity was essential for migration. ADAM 15, extracted from HMC membranes, was an active metalloproteinase, which degraded both type IV collagen and gelatin prepared from fibrillar collagen. Activity was inhibited by EDTA but not by phenylmethylsulfonyl fluoride. This is the first report of the potential of ADAM 15 for involvement in the restructuring of the mesangial matrix and in the migration of MC in disease. PMID- 12091382 TI - Nitrosative stress-induced apoptosis through inhibition of NF-kappa B. AB - Nitrosative stress produced by cytokines predisposes to apoptotic cell death. However, the molecular mechanism by which this occurs is not well understood. We have shown previously that nitric oxide (NO) regulates the activity of the anti apoptotic transcription factor NF-kappaB. Here we demonstrate that the inhibition of NF-kappaB by NO sensitizes A549 and Jurkat T cells to tumor necrosis factor alpha (TNFalpha)-induced apoptosis. The molecular basis of NF-kappaB inhibition is different in the two cell types. In A549 cells, NO functions at the nuclear level to inhibit NF-kappaB by S-nitrosylation. In Jurkat cells, NO inhibits the NF-kappaB activating pathway in the cytoplasm at a step proximal to the degradation of IkappaBalpha. The inhibition of NF-kappaB is reflected in the level of intracellular S-nitrosothiols, which are constitutively metabolized. These data suggest that NO can influence cell death by modulating NF-kappaB activity with the sites of inhibition being cell type-specific. The data also show that NO bioactivity regulates tumor necrosis factor-alpha signaling. PMID- 12091381 TI - Characterization of new cell permeable C3-like proteins that inactivate Rho and stimulate neurite outgrowth on inhibitory substrates. AB - The activation state of Rho is an important determinant of axon growth and regeneration in neurons. Axons can extend neurites on growth inhibitory substrates when Rho is inactivated by C3-ADP-ribosyltransferase (C3). We found by Rho-GTP pull-down assay that inhibitory substrates activate Rho. To inactivate Rho, scrape-loading of C3 was necessary because it does not freely enter cells. To overcome the poor permeability of C3, we made and characterized five new recombinant C3-like chimeric proteins designed to cross the cell membrane by receptor-independent mechanisms. These proteins were constructed by the addition of short transport peptides to the carboxyl-terminal of C3 and tested using a bioassay measuring neurite outgrowth of PC-12 cells plated on growth inhibitory substrates. All five constructs stimulated neurite outgrowth but with different dose-response profiles. Biochemical properties of the chimeric proteins were examined using C3-05, the most effective construct tested. Gel shift assays showed that C3-05 retained the ability to ADP-ribosylate Rho. Western blots and immunocytochemistry were used to verify the presence of C3 inside treated cells. C3-05 was also effective at promoting neurite outgrowth in primary neuronal cultures, as well as causing the disassembly of actin stress fibers and focal adhesions complexes in fibroblasts. These studies demonstrate that the new C3 like proteins are effective in delivering biologically active C3 into different cell types, thereby, inactivating Rho. PMID- 12091383 TI - Pore formation and function of phosphoporin PhoE of Escherichia coli are determined by the core sugar moiety of lipopolysaccharide. AB - The lipid matrix of the outer membrane of Gram-negative bacteria is an asymmetric bilayer composed of a phospholipid inner leaflet and a lipopolysaccharide outer leaflet. Incorporated into this lipid matrix are, among other macromolecules, the porins, which have a sieve-like function for the transport or exclusion of hydrophilic substances. It is known that a reduced amount of porins is found in the outer membrane of rough mutants as compared with wild-type bacteria. This observation was discussed to be caused by a reduced number of insertion sites in the former. We performed electrical measurements on reconstituted planar bilayers composed of lipopolysaccharide on one side and a phospholipid mixture on the other side using lipopolysaccharide from various rough mutant strains of Salmonella enterica serovar Minnesota. We found that pore formation by PhoE trimers that were added to the phospholipid side of the bilayers increased with the increasing length of the lipopolysaccharide core sugar moiety. These results allow us to conclude that the length of the sugar moiety of lipopolysaccharide is the parameter governing pore formation and that no particular insertion sites are required. Furthermore, we found that the voltage gating of the porin channels is strongly dependent on the composition of the lipid matrix. PMID- 12091384 TI - Protein kinase C-associated kinase (PKK) mediates Bcl10-independent NF-kappa B activation induced by phorbol ester. AB - Protein kinase C-associated kinase (PKK) is a recently described kinase of unknown function that was identified on the basis of its specific interaction with PKC beta. PKK contains N-terminal kinase and C-terminal ankyrin repeats domains linked to an intermediate region. Here we report that the kinase domain of PKK is highly homologous to that of two mediators of nuclear factor-kappa B (NF-kappa B) activation, RICK and RIP, but these related kinases have different C terminal domains for binding to upstream factors. We find that expression of PKK, like RICK and RIP, induces NF-kappa B activation. Mutational analysis revealed that the kinase domain of PKK is essential for NF-kappa B activation, whereas replacement of serine residues in the putative activation loop did not affect the ability of PKK to activate NF-kappa B. A catalytic inactive PKK mutant inhibited NF-kappa B activation induced by phorbol ester and Ca(2+)-ionophore, but it did not block that mediated by tumor necrosis factor alpha, interleukin-1 beta, or Nod1. Inhibition of NF-kappa B activation by dominant negative PKK was reverted by co-expression of PKC beta I, suggesting a functional association between PKK and PKC beta I. PKK-mediated NF-kappa B activation required IKK alpha and IKK beta but not IKK gamma, the regulatory subunit of the IKK complex. Moreover, NF kappa B activation induced by PKK was not inhibited by dominant negative Bimp1 and proceeded in the absence of Bcl10, two components of a recently described PKC signaling pathway. These results suggest that PKK is a member of the RICK/RIP family of kinases, which is involved in a PKC-activated NF-kappa B signaling pathway that is independent of Bcl10 and IKK gamma. PMID- 12091385 TI - Hyposialylation of integrins stimulates the activity of myeloid fibronectin receptors. AB - Despite numerous reports suggesting that beta(1) integrin receptors undergo differential glycosylation, the potential role of N-linked carbohydrates in modulating integrin function has been largely ignored. In the present study, we find that beta(1) integrins are differentially glycosylated during phorbol ester (PMA)-stimulated differentiation of myeloid cells along the monocyte/macrophage lineage. PMA treatment of two myeloid cell lines, U937 and THP-1, induces a down regulation in expression of the ST6Gal I sialyltransferase. Correspondingly, the beta(1) integrin subunit becomes hyposialylated, suggesting that the beta(1) integrin is a substrate for this enzyme. The expression of hyposialylated beta(1) integrin isoforms is temporally correlated with enhanced binding of myeloid cells to fibronectin, and, importantly, fibronectin binding is inhibited when the Golgi disrupter, brefeldin A, is used to block the expression of the hyposialylated form. Consistent with the observation that cells with hyposialylated integrins are more adhesive to fibronectin, we demonstrate that the enzymatic removal of sialic acid residues from purified alpha(5)beta(1) integrins stimulates fibronectin binding by these integrins. These data support the hypothesis that unsialylated beta(1) integrins are more adhesive to fibronectin, although desialylation of alpha(5) subunits could also contribute to increased fibronectin binding. Collectively our results suggest a novel mechanism for regulation of the beta(1) integrin family of cell adhesion receptors. PMID- 12091386 TI - p38 kinase regulates nitric oxide-induced apoptosis of articular chondrocytes by accumulating p53 via NFkappa B-dependent transcription and stabilization by serine 15 phosphorylation. AB - Nitric oxide (NO) during primary culture of articular chondrocytes causes apoptosis via p38 mitogen-activated protein kinase in association with elevation of p53 protein level, caspase-3 activation, and differentiation status. In this study, we characterized the molecular mechanism by which p38 kinase induces apoptosis through activation of p53. We report here that NO-induced activation of p38 kinase leads to activation of NFkappaB, which in turn induces transcription of the p53 gene. Activated p38 kinase also physically associates and phosphorylates the serine 15 residue of p53, which results in accumulation of p53 protein during NO-induced apoptosis. Ectopic expression of wild-type p53 enhanced NO-induced apoptosis, whereas expression of a dominant negative p53 blocked it, indicating that p53 plays an essential role in NO-induced apoptosis of chondrocytes. The increased accumulation of p53 caused expression of Bax, a pro apoptotic member of the Bcl-2 family that is known to cause apoptosis via release of cytochrome c and caspase activation. These results suggest that NO-activated p38 kinase activates p53 function in two different ways, transcriptional activation by NFkappaB and direct phosphorylation of p53 protein, leading to apoptosis of articular chondrocytes. PMID- 12091387 TI - The long and short isoforms of Ret function as independent signaling complexes. AB - Ret, the receptor tyrosine kinase for the glial cell line-derived neurotrophic factor family ligands (GFLs), is alternatively spliced to yield at least two isoforms, Ret9 and Ret51, which differ only in their C termini. To identify tyrosines in Ret that are autophosphorylation sites in neurons, we generated antibodies specific to phosphorylated Y905Ret, Y1015Ret, Y1062Ret, and Y1096Ret, all of which are autophosphorylated in cell lines. All four of these tyrosines in Ret became phosphorylated rapidly upon activation by GFLs in sympathetic neurons. These tyrosines remained phosphorylated in sympathetic neurons in the continued presence of GFLs, albeit at a lower level than immediately after GFL treatment. Comparison of GFL activation of Ret9 and Ret51 revealed that phosphorylation of Tyr(905) and Tyr(1062) was greater and more sustained in Ret9 as compared with Ret51. In contrast, Tyr(1015) was more highly phosphorylated over time in Ret51 than in Ret9. Surprisingly, Ret9 and Ret51 did not associate with each other in sympathetic neurons after glial cell line-derived neurotrophic factor stimulation, even though they share identical extracellular domains. Furthermore, the signaling complex associated with Ret9 was markedly different from the Ret51 associated signaling complex. Taken together, these data provide a biochemical basis for the dramatic functional differences between Ret9 and Ret 51 in vivo. PMID- 12091388 TI - Interaction of the c-Jun/JNK pathway and cyclin-dependent kinases in death of embryonic cortical neurons evoked by DNA damage. AB - DNA damage, an important initiator of neuronal death, has been implicated in numerous neurodegenerative conditions. We previously delineated several pathways that control embryonic cortical neuronal death evoked by the DNA-damaging agent, camptothecin. In this model, the tumor suppressor p53 and cyclin-dependent kinases (CDKs) are activated independently and cooperate to mediate the conserved death pathway. To further our understanding, we presently examined whether the c Jun/JNK pathway modulates death and whether this pathway is regulated by CDKs, p53, and Bax. We show that c-Jun/JNK is activated following DNA damage. Moreover, the c-Jun pathway is one mediator of death, because expression of dominant negative c-Jun and cdc42, and JNK pathway inhibitors are neuroprotective. Although previous evidences indicate that JNK3 is required for neuronal death under certain conditions, we show that JNK3 deficiency only partially mediates c Jun phosphorylation and its deficiency does not protect neurons from death. Interestingly, we provide evidence that CDK activity regulates c-Jun but does not affect upstream pathways that lead to JNK phosphorylation. Finally, c-Jun activation is independent of p53 and Bax. Accordingly, we propose that c-Jun is regulated by the JNK and CDK pathways and that both must be activated for efficient c-Jun activation to occur. PMID- 12091389 TI - Src-induced phosphorylation of caveolin-2 on tyrosine 19. Phospho-caveolin-2 (Tyr(P)19) is localized near focal adhesions, remains associated with lipid rafts/caveolae, but no longer forms a high molecular mass hetero-oligomer with caveolin-1. AB - Caveolin-2 is the least well studied member of the caveolin gene family. It is believed that caveolin-2 is an "accessory protein" that functions in conjunction with caveolin-1. At the level of the ER, caveolin-2 interacts with caveolin-1 to form a high molecular mass hetero-oligomeric complex that is targeted to lipid rafts and drives the formation of caveolae. However, caveolin-2 is not required for caveolae formation, implying that it may fulfill some unknown regulatory role. Here, we present the first evidence that caveolin-2 is a phosphoprotein. We show that caveolin-2 undergoes Src-induced phosphorylation on tyrosine 19. To study this phosphorylation event in vivo, we generated a novel phospho-specific antibody probe that only recognizes phosphocaveolin-2 (Tyr(P)(19)). We then used NIH-3T3 cells stably overexpressing c-Src to examine the localization and biochemical properties of phosphocaveolin-2 (Tyr(P)(19)). Our results indicate that phosphocaveolin-2 (Tyr(P)(19)) is localized near focal adhesions, remains associated with lipid rafts/caveolae, but no longer forms a high molecular mass hetero-oligomer with caveolin-1. Instead, phosphocaveolin-2 (Tyr(P)(19)) behaves as a monomer/dimer in velocity gradients. Thus, we conclude that the tyrosine phosphorylation of caveolin-2 (Tyr(P)(19)) may function as a signal that is recognized by the cellular machinery to induce the dissociation of caveolin-2 from caveolin-1 oligomers. We also demonstrate that (i) insulin-stimulation of adipocytes and (ii) integrin ligation of endothelial cells can both induce the tyrosine phosphorylation of caveolin-2 (Tyr(P)(19)). During integrin ligation, phosphocaveolin-2 (Tyr(P)(19)) co-localizes with activated FAK at focal adhesions. Thus, phosphocaveolin-2 (Tyr(P)(19)) may function as a docking site for Src homology domain-2 (SH2) domain containing proteins during signal transduction. In support of this notion, we identify several SH2 domain containing proteins, namely c-Src, NCK, and Ras-GAP, that interact with caveolin 2 in a phosphorylation-dependent manner. Furthermore, our co-immunoprecipitation experiments show that caveolin-2 and Ras-GAP are constitutively associated in c Src expressing NIH-3T3 cells, but not in untransfected NIH-3T3 cells. PMID- 12091390 TI - Silencing of transcription of the human luteinizing hormone receptor gene by histone deacetylase-mSin3A complex. AB - Modification of chromatin structure by histone acetylases and deacetylases is an important mechanism in modulation of eukaryotic gene transcription. The present study investigated regulation of the human luteinizing hormone receptor (hLHR) gene by histone deacetylases. Inhibition of histone deacetylases (HDACs) by trichostatin A (TSA) increased hLHR promoter activity by 40-fold in JAR cells and markedly elevated endogenous hLHR mRNA levels. Acetylated histones H3 and H4 accumulated in TSA-treated cells and associated predominantly with the hLHR promoter. Furthermore, TSA significantly enhanced the recruitment of RNA polymerase II to the promoter. One of the two Sp1 sites essential for basal promoter activity was identified as critical for the TSA effect, but the binding of Sp1/Sp3 to this site remained unchanged in the absence or presence of TSA. A multiprotein complex was recruited to the hLHR promoter via interaction with Sp1 and Sp3, in which HDAC1 and HDAC2 were docked directly to Sp1-bound DNA and indirectly to Sp3-bound DNA through RbAp48, while mSin3A interacted with both HDACs. HDAC1 and HDAC2 were shown to potently repress the hLHR gene transcription, and mSin3A potentiated the inhibition mediated by HDAC1. Our studies have demonstrated that the HDAC-mSin3A complex has an important role in the regulation of hLHR gene transcription by interaction with Sp1/Sp3 and by region-specific changes in histone acetylation and polymerase II recruitment within the hLHR promoter. PMID- 12091391 TI - A novel myc target gene, mina53, that is involved in cell proliferation. AB - Myc is a ubiquitous mediator of cell proliferation and can transactivate the expression of various genes through E-box sites. Here we report a novel gene, mina53 (Myc-induced nuclear antigen with a molecular mass of 53 kDa). The mina53 gene encodes a protein with a molecular weight of 53 kDa, which is localized in the nucleus and with part of the protein concentrated in the nucleolus. When serum-starved cells were activated by serum, the level of c-myc mRNA was elevated, and an increase in mina53 mRNA followed the elevation of c-myc mRNA. When expression of c-myc was reduced in human promyelocytic leukemia HL60 cells by phorbol 12-myristate 13-acetate, the expression of mina53 mRNA and protein was reduced. The expression of mina53 mRNA and Mina53 protein was induced by ectopic introduction of wild type c-Myc but not by a mutant c-Myc lacking the transactivation domain. When c-Myc in the c-MycER chimeric protein was activated, mina53 mRNA was increased, even in the presence of an inhibitor for protein synthesis. E-box sites are present in a region proximal to the transcription initiation sites of the mina53 gene. The gene expression from the mina53 promoter was elevated by c-Myc through E-box sites. c-Myc protein bound to the mina53 promoter region in vivo in HL60 cells in the proliferating phase but not after treatment of cells with phorbol 12-myristate 13-acetate. Specific inhibition of mina53 expression by an RNA interference method severely suppressed cell proliferation. Taken together, these results indicate that mina53 is a direct target gene of Myc, suggesting that mina53 is involved in mammalian cell proliferation. PMID- 12091392 TI - Bisindolylmaleimide IX facilitates tumor necrosis factor receptor family-mediated cell death and acts as an inhibitor of transcription. AB - Bisindolylmaleimides (Bis) were originally described as protein kinase C inhibitors. Several studies have shown that Bis potentiate tumor necrosis factor (TNF) receptor family-mediated apoptosis in lymphoid and dendritic cells, but the inhibition of protein kinase C cannot account for these effects (Zhou, T., Song, L., Yang, P., Wang, Z., Lui, D., and Jope, R. S. (1999) Nat. Med. 5, 42-48). We investigated the effect of four Bis derivatives (I, II, VIII, and IX) in human prostatic carcinoma cell lines and found that Bis IX was the most potent inducer of apoptosis under simultaneous treatment with TNF-alpha, agonistic anti-Fas monoclonal antibody, and TNF-related apoptosis-inducing ligand (TRAIL). Bis IX synergistically induced caspase activity in combination with apoptosis-inducing ligands and converted the phenotype of cell lines from apoptosis-resistant to sensitive. Bis IX induced p53 accumulation in LNCaP (lymph node carcinoma of prostate), which expresses wild-type p53 that was not accompanied by the induction of p53-responsive genes, p21/WAF1, and Mdm2. Moreover, the induction of p21/WAF1 and Mdm2 by doxorubicin was abrogated by simultaneous treatment with Bis IX. These effects apparently result from general inhibition of transcription by Bis IX. We have shown by Northern blot analysis that the transcription activity of the hygromycin gene after transient transfection of pcDNA3.1-Hygro plasmid in 293 and HeLa cells was inhibited by Bis IX in a dose-dependent manner. Moreover, DNA binding activity of Bis IX was prevented by actinomycin D, suggesting that actinomycin D and Bis IX have similar mechanisms of interaction with DNA. In addition, we found that actinomycin D and Bis IX induced caspase activity to the same extent during TRAIL-mediated apoptosis. In summary, these results suggest that Bis IX potentiates TNF receptor family-mediated cell death in part as an inhibitor of transcription. PMID- 12091393 TI - A rhodopsin mutant linked to autosomal dominant retinitis pigmentosa is prone to aggregate and interacts with the ubiquitin proteasome system. AB - The inherited retinal degenerations are typified by retinitis pigmentosa (RP), a heterogeneous group of inherited disorders that causes the destruction of photoreceptor cells, the retinal pigmented epithelium, and choroid. This group of blinding conditions affects over 1.5 million persons worldwide. Approximately 30 40% of human autosomal dominant (AD) RP is caused by dominantly inherited missense mutations in the rhodopsin gene. Here we show that P23H, the most frequent RP mutation in American patients, renders rhodopsin extremely prone to form high molecular weight oligomeric species in the cytoplasm of transfected cells. Aggregated P23H accumulates in aggresomes, which are pericentriolar inclusion bodies that require an intact microtubule cytoskeleton to form. Using fluorescence resonance energy transfer (FRET), we observe that P23H aggregates in the cytoplasm even at extremely low expression levels. Our data show that the P23H mutation destabilizes the protein and targets it for degradation by the ubiquitin proteasome system. P23H is stabilized by proteasome inhibitors and by co-expression of a dominant negative form of ubiquitin. We show that expression of P23H, but not wild-type rhodopsin, results in a generalized impairment of the ubiquitin proteasome system, suggesting a mechanism for photoreceptor degeneration that links RP to a broad class of neurodegenerative diseases. PMID- 12091394 TI - Apoptotic changes in the aged brain are triggered by interleukin-1beta-induced activation of p38 and reversed by treatment with eicosapentaenoic acid. AB - Among the several changes that occur in the aged brain is an increase in the concentration of the proinflammatory cytokine interleukin-1beta that is coupled with a deterioration in cell function. This study investigated the possibility that treatment with the polyunsaturated fatty acid eicosapentaenoic acid might prevent interleukin-1beta-induced deterioration in neuronal function. Assessment of four markers of apoptotic cell death, cytochrome c translocation, caspase-3 activation, poly(ADP-ribose) polymerase cleavage, and terminal dUTP nick-end staining, revealed an age-related increase in each of these measures, and the evidence presented indicates that treatment of aged rats with eicosapentaenoate reversed these changes as well as the accompanying increases in interleukin-1beta concentration and p38 activation. The data are consistent with the idea that activation of p38 plays a significant role in inducing the changes described since interleukin-1beta-induced activation of cytochrome c translocation and caspase-3 activation in cortical tissue in vitro were reversed by the p38 inhibitor SB203580. The age-related increases in interleukin-1beta concentration and p38 activation in cortex were mirrored by similar changes in hippocampus. These changes were coupled with an age-related deficit in long term potentiation in perforant path-granule cell synapses, while eicosapentaenoate treatment was associated with reversal of age-related changes in interleukin-1beta and p38 and with restoration of long term potentiation. PMID- 12091395 TI - Structural basis for the NAD-dependent deacetylase mechanism of Sir2. AB - The NAD-dependent histone/protein deacetylase activity of Sir2 (silent information regulator 2) accounts for its diverse biological roles including gene silencing, DNA damage repair, cell cycle regulation, and life span extension. We provide crystallographic evidence that 2'-O-acetyl ADP-ribose is the reaction product that is formed at the active site of Sir2 from the 2.6-A co-crystal structure of 2'-O-acetyl-ADP-ribose and Sir2 from Archaeoglobus fulgidus. In addition, we show that His-116 and Phe-159 play critical roles in the catalysis and substrate recognition. The conserved Ser-24 and Asp-101 contribute to the stability for NAD binding rather than being directly involved in the catalysis. The crystal structures of wild type and mutant derivatives of Sir2, in conjunction with biochemical analyses of the mutants, provide novel insights into the reaction mechanism of Sir2-mediated deacetylation. PMID- 12091396 TI - The GTPase Rap1 regulates phorbol 12-myristate 13-acetate-stimulated but not ligand-induced beta 1 integrin-dependent leukocyte adhesion. AB - Leukocyte migration from bloodstream to tissue requires rapid, coordinated regulation of integrin-dependent adhesion and de-adhesion. In a previous study we demonstrated that inhibition of protein geranylgeranylation inhibited phorbol ester-stimulated avidity modulation of beta(1) integrin in several leukocyte cell lines. Both RhoA and Rap1 require post-translational modification by geranylgeranylation for full function. In this report we identify Rap1, not RhoA, as a critical geranylgeranylated protein mediating phorbol ester-stimulated beta(1) and beta(2) integrin-dependent adhesion of Jurkat cells. Overexpression of the Rap1-specific GTPase-activating protein, SPA-1, or inactivated form of Rap1 (N17Rap1) blocked phorbol ester-stimulated adhesion of Jurkat cells to fibronectin (alpha(4)beta(1)) and ICAM-1 (alpha(L)beta(2)). With high concentrations of fibronectin as ligand, Jurkat cells adhered spontaneously without phorbol ester stimulation. Unlike the phorbol ester-stimulated adhesion, adhesion induced by high density ligand was not dependent upon Rap1 activation or actin cytoskeleton reorganization. Thus, the "inside-out" adhesion signal induced by phorbol ester and the "outside-in" signal induced by high density ligand involve different pathways. PMID- 12091397 TI - Nerve fiber layer splaying at vascular crossings. AB - PURPOSE: To examine the anatomic relationships of retinal blood vessels to the nerve fiber layer. METHODS: Retinal flat preparations from monkey and cat were stained with methylene blue dye and examined by light microscopy. RESULTS: A focal transition in nerve fiber bundle patterns occurs just above major retinal blood vessels. Bundles abandon a compact form and splay open just as they reach a major blood vessel. Maximum separation of axon fascicles occurs immediately above the blood vessel and the bundles reform after the crossing of the blood vessel is complete. In so doing, nerve fiber bundles temporarily thin and broaden, assuming the shape of an inverted hull. In contrast, at the retinal periphery, minor blood vessels and axon fascicles have no special relationship to each other. CONCLUSIONS: At major blood vessel crossings, a focal alteration in nerve fiber bundle anatomy takes place under which local deformability may be enhanced. This adaptation probably lessens the risk of injury to ganglion cell axons from vascular compression. In addition, deflection of the nerve fiber pathway obviates the need for major blood vessels to bend or kink to achieve a crossing, thus avoiding turbulent flow. PMID- 12091399 TI - Characterization of the macrophages associated with the tunica vasculosa lentis of the rat eye. AB - PURPOSE: The tunica vasculosa lentis (TVL) is a transient vascular network surrounding the developing lens that regresses prenatally in humans and postnatally in rodents. Macrophage-like cells, sometimes referred to as hyalocytes, have been postulated to play a role in the regression of this vascular system; however, the precise identity of these cells is still unclear. The aim of the present investigation was to provide phenotypic data on these cells in combination with their three-dimensional distribution on the lens surface during the period when regression of the TVL is taking place. To this end the authors have used a novel combination of silver-enhanced immunogold labeling and environmental scanning electron microscopy (ESEM). METHODS: The eyes of Wistar rats at various pre- and postnatal ages (E20, postnatal days [D]0, 2, 5, 7, and 10) were studied by either conventional scanning electron microscopy (SEM) or ESEM. The latter was used to study specimens that had been incubated with various antileukocyte monoclonal antibodies (ED1, ED2, Ox6, Ox42), followed by immunolabeling with gold-conjugated secondary antibody visualized by silver enhancement. RESULTS: Conventional SEM of the developing lens revealed a pattern of radial and interconnecting vessels in the TVL similar to previous studies. In addition, large numbers of cells with the morphologic characteristics of macrophages were noted on the lens surface closely associated with the vessels. The gradual attenuation and regression of vessels was noted over the course of the time period investigated. Immunolabeled specimens examined by ESEM revealed that most of the macrophage-like cells were indeed ED1(+) and ED2(+) (both pan macrophage markers) and MHC class II(-) (Ox6) and CD11b/18(-) (Ox42): a phenotype characteristic of macrophages. This phenotype altered little between E20 and D10. CONCLUSIONS: The cells surrounding the developing lens that are postulated to play a role in regression of the TVL have the morphologic and immunophenotypic characteristics of resident tissue macrophages similar to those previously identified in the adult rodent uveal tract and the vitreous (hyalocytes). This phenotype differs from that of dendritic cells and microglia; however, it is postulated that lens-associated macrophages are ideally located to act as a source of retinal microglia after completion of their role in TVL regression. PMID- 12091400 TI - A novel locus for autosomal dominant nuclear cataract mapped to chromosome 2p12 in a Pakistani family. AB - PURPOSE: To map the disease locus in a four-generation, consanguineous Pakistani family affected by autosomal dominant congenital nuclear cataract (adNCat). All affected individuals had early onset of bilateral nuclear cataract. METHODS: Genomic DNA from family members was typed for alleles at more than 300 known polymorphic genetic markers by polymerase chain reaction. The lod scores were calculated by using two-point linkage analysis of the genotyping data. RESULTS: The maximum lod score, 4.05, was obtained for the marker D2S2333. Proximal and distal crossovers were observed with markers D2S286 and D2S1790, respectively. These crossovers define the critical disease locus to an interval of approximately 9 centimorgans (cM). CONCLUSIONS: Linkage analysis identified a novel locus for adNCat on chromosome 2p12 in a Pakistani family. A genome database analysis of the target interval is being undertaken to identify candidate gene(s) for the disease. PMID- 12091398 TI - The time course of changes in mRNA levels in tree shrew sclera during induced myopia and recovery. AB - PURPOSE: In tree shrews, visual form deprivation produces increased axial elongation of the deprived eye and a myopic shift in refractive state. A change in scleral extensibility (creep rate) is closely associated with the change in axial elongation rate. These effects may be due to scleral tissue remodeling produced by a change in scleral gene expression. In this study, the authors investigated the time course of changes in scleral mRNA levels for selected proteins during the development of form deprivation myopia and during recovery, to determine which, if any, are temporally associated with changes in scleral extensibility and axial elongation rate. METHODS: Competitive RT-PCR was used to measure the levels of mRNA for structural proteins (collagen [alpha1(I) chain], decorin core protein), degradative enzymes (MMP-2 [gelatinase-A], MMP-3 [stromelysin-1]), and a tissue inhibitor of metalloproteinase (TIMP-1) in the scleras of tree shrews that had been subjected to 1, 2, 4, or 11 days of monocular form deprivation (MD) or 11 days of MD followed by 2 or 4 days of recovery produced by removal of the MD. Four groups of normal animals provided age-matched normal data. RESULTS: Compared with the control eyes, deprived-eye MMP-2 mRNA levels were higher and MMP-3 levels were lower after 4 days of MD. Deprived-eye collagen mRNA levels were lower than control eye levels after 11 days of MD. The differential effects produced by MD were absent after 2 days of recovery and generally were reversed after 4 days. Decorin mRNA levels in the deprived and control eyes were not significantly different during either MD or recovery. During MD, mRNA levels for collagen, MMP-3, and TIMP-1 decreased in both the deprived and control eyes, compared with age-matched normal eyes. The binocular changes in collagen and TIMP-1 mRNA levels and the differential changes in MMP-2 and MMP-3 levels were detected at least as early as axial, refractive, and creep rate changes. CONCLUSIONS: The up- and downregulation of the specific mRNAs studied, on a time course similar to that for physical changes in the sclera, suggests that modulation of gene expression by the visual environment may produce scleral remodeling and changes in scleral creep rate during the development of form deprivation myopia and recovery. PMID- 12091401 TI - Photoreactivity of aged human RPE melanosomes: a comparison with lipofuscin. AB - PURPOSE: To determine whether aging is accompanied by changes in aerobic photoreactivity of retinal pigment epithelial (RPE) melanosomes isolated from human donors of different ages, and to compare the photoreactivity of aged melanosomes with that of RPE lipofuscin. METHODS: Human RPE pigment granules were isolated from RPE cells pooled into groups according to the age of the donors. Photoreactivity was determined by blue-light-induced oxygen uptake and photogeneration of reactive oxygen species. Short-lived radical intermediates were detected by spin-trapping, hydrogen peroxide by an oxidase electrode, singlet oxygen by cholesterol assay, and lipid hydroperoxides by iodometric assay. RESULTS: Blue-light photoexcitation of melanosomes resulted in age-related increases in both oxygen uptake and the accumulation of superoxide anion spin adducts. The efficiencies of these processes, however, were still significantly lower than that induced by photoexcited lipofuscin. During irradiation of melanosomes, a substantial amount of oxygen was converted into hydrogen peroxide, whereas for lipofuscin, hydrogen peroxide accounted for not more than 3% of oxygen consumed. In contrast to lipofuscin, photoexcited melanosomes did not substantially increase the rate of oxidative reactions in the presence of polyunsaturated lipids or albumin. However, oxygen uptake was significantly elevated in the presence of ascorbate. Thus, the rate of photo-induced oxygen uptake in samples containing both ascorbate and melanosomes approached that observed in lipofuscin samples. CONCLUSIONS: Blue-light-induced photoreactivity of melanosomes increases with age, perhaps providing a source of reactive oxygen species and leading to depletion of vital cellular reductants, which, together with lipofuscin, may contribute to cellular dysfunction. PMID- 12091402 TI - Retinal voltage-dependent anion channel: characterization and cellular localization. AB - PURPOSE: To characterize and localize retinal voltage-dependent anion channel (VDAC) and to understand its possible contribution to mitochondrial function and dysfunction. METHODS: VDAC was characterized by a method involving purification from isolated mitochondria and reconstitution into a planar lipid bilayer (PLB). The permeability transition pore (PTP) was monitored by Ca(2+) accumulation in isolated mitochondria and swelling of mitochondria. Localization was studied by immunocytochemistry and in situ hybridization. RESULTS: Retinal VDACs exhibited the electrophysiological fingerprint of the VDAC superfamily. It had a maximal chord conductance of 3.7 +/- 0.1 nanosiemens (nS) in 1 M NaCl, and a voltage dependent conductance that was highest at transmembrane potential close to zero. It was modulated by glutamate, which decreased the channel's open probability, and by La(3+) and ruthenium amine binuclear complex (Ru360), which closed the channel. Energized and freshly prepared retinal mitochondria accumulated Ca(2+) that is inhibited by La(3+) ruthenium red and Ru360. Subsequent to Ca(2+) accumulation, mitochondria released the accumulated Ca(2+), probably through activation of the PTP. Ru360 inhibited Ca(2+) release and mitochondrial swelling. VDAC was present in mitochondria of all retinal cell types: photoreceptor, bipolar, horizontal, amacrine, and ganglion cells. Most cells primarily expressed VDAC-1, but they also expressed VDAC-2 and -3. CONCLUSIONS: These results suggest that VDAC is involved in PTP activity and/or regulation and thus is an important player in retinal degeneration associated with PTP-mediated mitochondrial dysfunction. PMID- 12091403 TI - Associations of human crystalline lens retrodots and waterclefts with visual impairment: an observational study. AB - PURPOSE: To investigate the relationships between visual acuity, contrast sensitivity, and 11 clinicopathologic classes of opacity in the human crystalline lens. METHODS: The Somerset and Avon Eye Study is an observational population study of age-related sight-threatening eye disease, based in Bristol, UK. After excluding eyes with other visually relevant disease, data from 902 individuals aged 55 years or older were analyzed. The associations of lens features (posterior subcapsular cataract, nuclear color, nuclear white scatter, cortical spokes, anterior subcapsular cataract, vacuoles, waterclefts, coronary flakes, focal dots, retrodots, fiber folds), with refracted log minimum angle of resolution (MAR) distance acuity and Pelli-Robson contrast sensitivity, were investigated. Multivariable linear regression models using data from both eyes and taking account of the intraclass correlation between eyes were used for analysis, with the lens features and age included as potential explanatory variables. RESULTS: As anticipated from earlier studies, posterior subcapsular, nuclear, and cortical cataracts were associated with visual impairment. In addition, retrodots were strongly and independently associated in the multivariable models with both impaired visual acuity (P < 0.001) and contrast sensitivity (P < 0.001). Waterclefts were strongly associated with impaired visual acuity (P < 0.001). CONCLUSIONS: Retrodots and waterclefts are associated with visual impairment. A causal relationship between these lens features and retinal image degradation is plausible. PMID- 12091404 TI - A retrospective study of myopia progression in adult contact lens wearers. AB - PURPOSE: To study retrospectively the frequency of myopia progression and risk factors for progression in a sample of adult contact lens wearers. METHODS: From a database of 815 soft contact lens wearers, patients were identified whose age was between 20 and 40 years, who had at least -0.50 D spherical equivalent of myopia in both eyes, three or more refractions, and > or =5 years of follow-up. Only data from the right eye were used. Progression was defined as an increase of at least -1.00 D over 5 years. Subjects were also asked to complete a questionnaire regarding their ocular history, demographics, family history, and the amount of time spent performing different tasks at home and at work. RESULTS: Two hundred ninety-one subjects met the eligibility criteria with a mean baseline refractive error of -3.29 +/- 1.92 D and a mean age of 28.5 +/- 5.0 years. Of these, 21.3% progressed by at least -1.00 D over the 5-year period. The 5-year rate of progression decreased with increasing age (chi(2) = 12.44, P = 0.006). One hundred ninety-seven subjects (67.6%) completed and returned questionnaires. "Progressors" (N = 41) did not differ from "nonprogressors" (N = 156) in terms of hours per day spent reading and writing, computer use, education level, family history of myopia, age of onset of myopia, and contact lens wear. CONCLUSIONS: In this database of soft contact lens wearers, myopia progression was common for subjects in their twenties and less common for those in their thirties. PMID- 12091405 TI - An evaluation of cultivated corneal limbal epithelial cells, using cell suspension culture. AB - PURPOSE: A previous report has described an ocular surface reconstruction method involving the use of cultivated corneal epithelium derived from limbal explants. In the current study, a new culture system was developed involving the in vitro propagation on amniotic membrane (AM) of epithelial cells from enzymatically dissociated limbal epithelium. The purpose of this new method is to produce a cultivated epithelial cell layer that contains stem cells and that is superior to explanted cultivated epithelium. The new cell-suspension technique was compared with the existing explant method. METHODS: Limbal epithelial cells were dissociated from donor eyes by dispase and seeded on the denuded AM. Small pieces of limbal epithelium were also cultured on denuded AM as explant cultures. The cultivated epithelium was examined by electron microscopy and immunohistochemistry for cornea-specific keratins (K3 and K12). RESULTS: Both cell-suspension and explant culture methods produced a healthy epithelial cell layer. The cell-suspension culture had significantly (P < 0.001) more desmosomal junctions between the explant-cultured basal cells. In addition, the intercellular spaces between the cell-suspension's basal cells were significantly (P < 0.001) smaller than those between the explant-cultured basal cells. Both types of cultivated epithelium showed positive expression of K3 and K12 keratins. In the cell-suspension culture, expression of K3 and K12 keratins was more prominent in the superficial cells. CONCLUSIONS: Corneal epithelial cells were successfully regenerated in vitro by a cell-suspension culture system. The suspension-cultured epithelium must include some stem cells and morphologically is significantly superior to explant-cultured epithelium. Thus, this new technique is potentially more suitable for cultivated corneal limbal epithelial transplantation. PMID- 12091406 TI - Mitogenic and antiapoptotic effects of various growth factors on human corneal fibroblasts. AB - PURPOSE: To investigate both the effects of various growth factors on the proliferation of human corneal fibroblasts and the abilities of these factors to protect the cells from apoptosis. METHODS: Cultured human corneal fibroblasts were incubated separately with 11 different growth factors whose receptors are expressed by these cells. Cell proliferation was evaluated by measurement of [(3)H]thymidine incorporation. The activation of the protein kinase Akt, which plays an important role in antiapoptotic signaling, was assessed by immunoblot analysis with antibodies specific for a phosphorylated form of the enzyme. Apoptosis was quantitated by the TdT-mediated dUTP-biotin nick-end labeling (TUNEL) assay. RESULTS: Of the 11 growth factors examined, platelet-derived growth factor, insulin, insulin-like growth factors-1 and -2, and epidermal growth factor, each stimulated the proliferation of corneal fibroblasts, induced the activation of Akt in these cells, and protected them from apoptosis induced by sodium nitroprusside (SNP). Basic fibroblast growth factor, keratinocyte growth factor, nerve growth factor, and hepatocyte growth factor stimulated cell proliferation but did not induce Akt activation or protect the cells from SNP induced apoptosis. Transforming growth factor-beta1 and -beta2 did not stimulate proliferation and had no effect on Akt activity or on SNP-induced apoptosis. CONCLUSIONS: In terms of their modulatory effects on the proliferation and apoptosis of human corneal fibroblasts, the 11 growth factors examined can be classified into three groups. These growth factors may both contribute to maintenance of the cornea and coordinate the proliferative and apoptotic responses of corneal fibroblasts during corneal wound healing. PMID- 12091407 TI - Lymphatic vessels in vascularized human corneas: immunohistochemical investigation using LYVE-1 and podoplanin. AB - PURPOSE: To determine whether lymphatic vessels exist in vascularized human corneas, by using immunohistochemistry with novel markers for lymphatic endothelium. METHODS: Human corneas exhibiting neovascularization secondary to keratitis, transplant rejection, trauma, and limbal insufficiency (n = 21) were assessed for lymphatic vessel content by conventional transmission electron microscopy and by immunostaining and immunoelectron microscopy with antibodies specific for the lymphatic endothelial markers, lymphatic vessel endothelial hyaluronan receptor (LYVE-1) and the 38-kDa integral membrane glycoprotein podoplanin. In addition, corneas were stained for the lymphangiogenic growth factor VEGF-C, and its receptor VEGFR3 by immunohistochemistry and in situ RNA hybridization, respectively. RESULTS: Thin-walled, erythrocyte-free vessels staining with lymphatic markers (LYVE-1 and podoplanin) were found to constitute 8% of all vessels, to be more common in the early course of neovascularization, to be always associated with blood vessels and stromal inflammatory cells, and to correlate significantly with the degree of corneal hemangiogenesis (r = 0.6; P = 0.005). VEGF-C, VEGFR3, podoplanin, and LYVE-1 colocalized on the endothelial lining of lymphatic vessels. With immunogold labeling, LYVE-1 and podoplanin antigen were found on endothelial cells lining vessels with ultrastructural features of lymph vessels. CONCLUSIONS: Immunohistochemistry with novel lymph endothelium markers and ultrastructural analyses indicate the existence of lymphatic vessels in vascularized human corneas. Human corneal lymphangiogenesis appears to be correlated with the degree of corneal hemangiogenesis and may at least partially be mediated by VEGF-C and its receptor VEGFR3. PMID- 12091408 TI - The effect of chronic corneal epithelial debridement on epithelial and stromal morphology in dogs. AB - PURPOSE: To determine the effect of chronic corneal epithelial debridement on epithelial and stromal morphology and extracellular matrix components, and to compare those changes to those in spontaneous chronic corneal epithelial defects (SCCED) in dogs. METHODS: Axial corneal epithelial wounds, 10 mm in diameter, were created weekly for 8 weeks in five normal adult laboratory beagles. Slit lamp biomicroscopy and corneal pachymetry were performed weekly before wounding. Three days after the last debridement the dogs were killed humanely, and corneas were processed for light and electron microscopy and immunohistochemistry for collagen IV, collagen VII, fibronectin, and laminin. RESULTS: No significant changes in corneal thickness were found. All samples demonstrated epithelial dysmaturation adjacent to the wound edge, and, in four of five, a narrow zone of nonadherent epithelium formed adjacent to the exposed stroma. All samples had a stromal acellular zone in the area of the defect and continuing for a short distance under the adjacent attached epithelium. Experimentally wounded dogs did not form the superficial hyaline acellular lamina found in 92% of dogs with SCCED. Laminin, collagen IV, and fibronectin were present on the stromal surface in all samples, and collagen VII was present in four of five samples. Transmission electron microscopy (TEM) demonstrated the presence of basement membrane on the surface of the exposed stroma. CONCLUSIONS: Epithelial changes are similar between experimentally wounded dogs and dogs with SCCED. The stromal acellular zone that forms in experimentally wounded dogs is distinct from the hyaline lamina observed in dogs with SCCED. The difference in the acellular stromal layers between chronically wounded dogs and dogs with SCCED may be of relevance to our understanding of the pathophysiology of persistent epithelial defects. PMID- 12091409 TI - Microarray analysis of corneal fibroblast gene expression after interleukin-1 treatment. AB - PURPOSE: To identify changes in gene expression in human corneal fibroblasts after exposure to interleukin-1alpha. METHODS: RNA was isolated from cultured human corneal fibroblasts after treatment with interleukin-1alpha and subjected to DNA microarray analysis. Changes in gene expression were determined by comparison with untreated cells in three independent experiments after a Bayesian statistical analysis of variance. RESULTS: Changes in gene expression were reproducibly observed in 165 genes representing previously identified and novel chemokines, matrix molecules, membrane receptors, angiogenic mediators, and transcription factors that correlated with pathophysiological responses to inflammation. Dramatic increases in gene expression were observed with exodus-1 (CCL20), MMP-12, and RhoA. CONCLUSIONS: DNA microarray analysis of the corneal fibroblast response to interleukin-1alpha provides important insight into modeling changes in gene expression and suggests novel therapeutic targets for the control of corneal inflammation. PMID- 12091410 TI - Mechanisms of mitotic inhibition in corneal endothelium: contact inhibition and TGF-beta2. AB - PURPOSE: Contact inhibition has been implicated as an important antiproliferative mechanism in developing and mature corneal endothelium. Although exogenous TGF beta2 and TGF-beta2 in aqueous humor suppress S-phase entry in cultured rat corneal endothelial cells, it is not known whether TGF-beta2 contributes to the mitotic inhibition that occurs during in vivo endothelial development. TGF-beta receptors I, II, and III must be coexpressed for a TGF-beta2-induced intracellular signal to be transmitted. The current study was conducted to determine whether TGF-beta2 contributes to mitotic inhibition during endothelial development, by investigating when these receptors become coexpressed in the endothelium of neonatal rats. Cyclin-dependent kinase inhibitors (CKIs), such as p27kip1 and p15INK4b, help mediate mitotic inhibition in other cell types. The role of CKIs in inhibiting proliferation in corneal endothelium was examined by first determining the kinetics of p27kip1 expression in neonatal rat corneal endothelium. Studies were then extended to cultured cells to more directly compare the effects of TGF-beta2 and cell-cell contact on the relative protein and mRNA expression of the CKIs, p27kip1, and p15INK4b. METHODS: Immunocytochemistry (ICC) detected TGF-beta receptors I, II, and III (RI, RII, RIII, respectively) in the endothelium of rat corneas on postnatal days 1, 10, and 21, and in adult (3-month-old) rats. ICC for p27kip1 was conducted on postnatal days 1, 7, 14, and 21. Samples were taken for p27kip1 RT-PCR on postnatal days 7, 14, and 21 and from adult rats. The effect of TGF-beta2 on p27kip1 and p15INK4b expression was determined in G(0)-phase synchronized subconfluent rat corneal endothelial cells incubated for 24 hours in 10% serum +/ 5 ng/mL TGF-beta2. CKI expression was also examined in fully confluent cultures. RT-PCR and Western blot analysis detected p27kip1 and p15INK4b mRNA and protein expression, respectively. The effect of releasing cells from cell-cell contact on proliferation and p27kip1 protein expression was studied in confluent cultures treated for 1 hour with and without 2.0 mg/mL di-sodium EDTA and then maintained for 24 hours in 10% serum. Cultures were then either fixed for ICC of Ki67, a marker of actively cycling cells, or extracted for Western blot determination of p27kip1 protein. RESULTS: Positive staining for TGF-beta RIII was detected on postnatal day 10, and staining for RI and RII was detected on postnatal day 21. The endothelium stained positively for p27kip1 on postnatal day 1 and thereafter, and p27kip1 PCR product was detectable at the earliest time point tested (postnatal day 7). In cultured cells, TGF-beta2 and cell-cell contact had relatively little effect on expression of p27kip1 or p15INK4b mRNA. TGF-beta2 lowered the levels of both proteins, but p27kip1 remained at a higher level than p15INK4b. In confluent cultures, p15INK4b protein was reduced; however, p27kip1 protein levels increased 20-fold. Positive staining for Ki67 was detected, and p27kip1 protein levels substantially decreased in EDTA-treated confluent cells compared with the untreated control. CONCLUSIONS: Previous studies from this laboratory showed that corneal endothelial cell proliferation ceases in neonatal rat by postnatal day 13. This timing correlated with the formation of stable cell cell contacts, implicating contact inhibition as an important mechanism of growth arrest during endothelial development. The current studies showed that coexpression of TGF-beta RI, RII, and RIII occurred too late for TGF-beta2 to have a significant role in inhibiting proliferation during endothelial development. Studies in cultured cells suggest that p27kip1 mediates inhibition of proliferation induced by TGF-beta2, although the response to this cytokine was relatively weak. ICC and RT-PCR of p27kip1 in neonatal endothelium and RT-PCR and Western blot studies in cultured cells indicate that contact inhibition is mediated, in large part, through the activity of p27kip1. These results, together with previous data from this laboratory, strongly suggest that contact inhibition is an important mechanism responsible for inducing cell cycle arrest during corneal endothelial development and for maintaining the mature monolayer in a nonproliferative state. In both cases, contact-induced inhibition is mediated, at least in part, by p27kip1. TGF-beta2 appears not to induce mitotic arrest in the developing endothelium, but may function to maintain the mature endothelium in a nonreplicative state should cell-cell contact be lost in the monolayer. PMID- 12091411 TI - Recovery of endothelial function after vitrification of cornea at -110 degrees C. AB - PURPOSE: To determine whether endothelial function is retained after ice-free cryopreservation of cornea by vitrification at -110 degrees C. METHODS: Rabbit corneas, mounted on support rings, were exposed to a solution containing 6.8 M propane-1,2-diol (PROH) and cooled at approximately 7 degrees C/min to -110 degrees C, which was below the glass transition temperature (T(g)) of the solution. After rewarming at approximately 12 degrees C/min and removal of the PROH, endothelial function was assessed by monitoring corneal thickness during perfusion at 34 degrees C. RESULTS: Addition and removal of 6.8 M PROH without cooling to -110 degrees C did not markedly impair endothelial function, although corneas were thicker than control samples. There was no visible crystallization of ice during cooling to -110 degrees C; but a few small, discrete sites of crystallization remote from the endothelium, were observed during warming. After removal of the PROH, corneas approximately doubled in thickness during the first 3 hours of perfusion, but they then started to thin, which suggested active control of stromal hydration by the endothelium. This was confirmed in a further set of experiments by removal of bicarbonate ions from the perfusate at this point, which resulted in further swelling at +58 +/- 2 microm/hour (SD; n = 4). Restoring bicarbonate to the perfusate halted this swelling, and the corneas then thinned at -13 +/- 2 microm/hour (n = 4). Morphologically, staining with trypan blue and alizarin red S showed an apparently intact endothelial monolayer. CONCLUSIONS: Rabbit corneal endothelium tolerated exposure to 6.8 M PROH, and endothelial function was evident after vitrification at -110 degrees C. Preliminary morphologic results with vitrified human cornea also showed retention of endothelium. PMID- 12091412 TI - UV absorbance of the human cornea in the 240- to 400-nm range. AB - PURPOSE: To determine the UV absorbance of the corneal layers (epithelium, Bowman layer, stroma) in the 240- to 400-nm range. METHODS: Consecutive slices (100 microm) of human cadaveric corneas were cut, and the UV absorbance of each sample was determined in a scanning spectrophotometer. In some cases the epithelium was scraped off and its absorbance measured separately. RESULTS: The investigation of the UV-B absorption of consecutive corneal slices revealed evidence that UV-B absorption is 1.8 times higher in the anterior 100 microm of the human cornea than in the posterior layers. The UV absorbance of the posterior layers was uniform, showing no further structural dependence. The epithelium and Bowman layer are both effective absorbers of UV-B radiation. CONCLUSIONS: These results suggest that the anterior corneal layers are particularly important in preventing damage by UV-B radiation. PMID- 12091413 TI - Incomitant strabismus associated with instability of rectus pulleys. AB - PURPOSE: Connective tissue pulleys serve as functional mechanical origins of the extraocular muscles (EOMs) and are normally stable relative to the orbit during gaze shifts. This study evaluated pulley stability in incomitant strabismus. METHODS: Contiguous 2- or 3-mm thick magnetic resonance images (MRIs) perpendicular to the orbital axis spanned the anteroposterior extents of 12 orbits of six patients with incomitant strabismus. Imaging was performed in central gaze, supraduction, infraduction, abduction, and adduction. Rectus EOM paths were defined by their area centroids and plotted in a normalized, oculocentric coordinate system. Paths of EOMs ran toward the pulleys. Sharp EOM path inflections in secondary gaze indicated pulley locations in three dimensions. RESULTS: MRI revealed substantial inferior shift of the lateral rectus (LR) pulley of up to 1 mm during vertical gaze shifts in patients with axial high myopia and a posterior shift from abduction to adduction in simulated Brown syndrome. There was substantial LR pulley shift opposite the direction of vertical gaze in a subject with X-pattern exotropia who had undergone repeated LR surgery. The medial rectus (MR) pulley shifted inferiorly with gaze elevation in Marfan syndrome. Pulley instability was associated with significantly increased globe translation during gaze shifts. CONCLUSIONS: Pulley instability, resulting in EOM sideslip during ductions, occurs in some cases of incomitant strabismus. Resultant patterns of strabismus may depend on static pulley positions, pulley instability, and coexisting globe translation that alters pulley locations relative to the globe. Translational instability of pulleys and the globe could produce abnormalities in actions of otherwise normal EOMs, and connective tissue disorders causing these instabilities should be considered as potential causes of strabismus. PMID- 12091415 TI - Adaptive modification of disparity vergence components: an independent component analysis study. AB - PURPOSE: Although a disparity vergence stimulus produces a smooth exponential like response, considerable experimental evidence indicates that it is the product of at least two motor components: a pulselike transient component and a steplike sustained component. Recently, a new application of independent component analysis (ICA) has been used to decompose the vergence step response into these underlying components. Other recent experiments have shown that the vergence system is capable of rapidly modifying its dynamic characteristics (short-term adaptation) when exposed to specially designed "adapting" stimuli. Adapted responses were characterized by faster dynamics, often featuring large overshoots. In this study, ICA was used to examine changes in the underlying components produced by dynamic adaptation. METHODS: Disparity vergence eye movements in response to identical step stimuli were obtained from four subjects both in normal (baseline) conditions and after modification by adaptive training stimuli. ICA-based component decomposition was applied to vergence step-response data sets in both normal and adapted conditions to estimate, and compare activation patterns of the two underlying components. RESULTS: An eigenvector analysis indicated that both normal and adapted vergence responses contained two major components. ICA analysis showed that the enhanced dynamics seen in adapted responses was due to an increase in pulse component amplitude. In addition, the step component of adapted responses often showed double-step behavior in the later portion of the response. Finally, the magnitude of adaptation appeared to be related to the unadapted response dynamics. CONCLUSIONS: The adaptive process does not evoke additional components, but modifies the two components that are present under normal conditions. Double steps seen in the step component were attributed to an interaction between pulse and step neural mechanisms. The generation of an enhanced pulse component interfered with the production of the step component. Under this scenario, the reduced initial-step component was then compensated by the generation of a second-step component, probably mediated by an internal feedback mechanism. PMID- 12091414 TI - Active pulleys: magnetic resonance imaging of rectus muscle paths in tertiary gazes. AB - PURPOSE: The orbital layer of each rectus extraocular muscle (EOM) inserts on connective tissue, and the global layer inserts on the eyeball. The active-pulley hypothesis (APH) proposes that a condensation of this connective tissue constitutes a pulley serving as the functional origin of the rectus EOM, and that this pulley makes coordinated, gaze-related translations along the EOM axis to implement a linear ocular motor plant. This study was designed to measure gaze related shifts in EOM pulley locations. METHODS: Magnetic resonance imaging (MRI) was performed in eight normal volunteers in 2-mm thickness coronal planes perpendicular to the orbital axis for nine cardinal gaze directions. Intravenous gadodiamide contrast was administered to define EOM tendons anterior to the globe equator. Paths of EOMs, defined by their area centroids, were transformed into an oculocentric coordinate system. Sharp inflections in EOM paths in secondary and tertiary gaze positions defined pulley locations which were then correlated with gaze direction and compared with theoretical predictions. RESULTS: Rectus pulley positions were consistent with a central primary position. In tertiary gaze positions, each of the four rectus pulleys translated posteriorly with EOM contraction and anteriorly with EOM relaxation by a significant (P < 0.02) amount predicted by the APH, but more than 100 times greater than the translation predicted by a passive pulley model. CONCLUSIONS: The APH prediction of coordinated anteroposterior shifting of EOM pulleys with gaze is quantitatively supported by changes in EOM path inflections among tertiary-gaze positions. Human rectus pulleys move to shift the ocular rotational axis to attain commutative behavior of the ocular motor plant. PMID- 12091416 TI - Effect of hyaluronic acid on intraocular pressure in rats. AB - PURPOSE: To study the effect of acute or chronic intracameral injection of hyaluronic acid on intraocular pressure (IOP) in rats. METHODS: Acute or chronic injections of hyaluronic acid were performed unilaterally in the rat eye's anterior chamber, whereas the contralateral eye was injected with saline solution. IOP was assessed daily or weekly by a tonometer in conscious rats. IOP was also assessed in both experimental groups at several intervals during the light-dark cycle. RESULTS: A single injection of hyaluronic acid induced an increase of IOP that lasted for 8 days (P < 0.01), whereas its chronic administration during 9 weeks induced a significant and sustained increase in IOP, compared with the eye injected with vehicle (P < 0.01). This hyaluronic acid induced hypertension was significantly decreased by the application of 1 drop of brimonidine (0.2%), latanoprost (0.005%), or timolol (0.5%). Significant daily variations of IOP were observed in both control and hyaluronic acid-injected eyes, peaking during the dark phase (P < 0.001, ANOVA). CONCLUSIONS: These results suggest that the intracameral administration of hyaluronic acid could be a model of ocular hypertension in rats. PMID- 12091417 TI - Identification of the mouse uveoscleral outflow pathway using fluorescent dextran. AB - PURPOSE: This study was undertaken to assess directly whether there is uveoscleral outflow in the mouse eye by monitoring the movement of intracamerally injected fluorescent dextran. METHODS: After anesthesia, NIH Swiss mice received intracameral injection of 1.5 microL of 0.2 pg/microL 70-kDa dextran conjugated to tetramethyl-rhodamine and to lysine. After survival times of 10, 20, 60, and 120 minutes, the experiments were terminated by transcardial perfusion with 2% paraformaldehyde. The eyes were enucleated and embedded in paraffin, and sections were prepared. These sections were then analyzed by fluorescence microscopy. RESULTS: Fluorescent tracer in the eyes of animals that survived for 10 minutes was prominent in the iris root and ciliary processes and was of moderate intensity in the adjacent sclera. Moderate intensity fluorescence also was observed in the trabecular meshwork and adjacent cornea. At 20 minutes, intense fluorescence was observed in the ciliary processes and the ciliary muscle. This fluorescence in the ciliary muscle extended from the posterior edge of the ciliary muscle's tail into the anterior choroid. At 60 minutes, the fluorescence in the choroid extended to the equator and adjacent sclera. The intensity of the fluorescence within the ciliary processes of these eyes was substantially reduced when compared with the 20-minute-survival eyes. At 120 minutes, label was observed only within trabecular meshwork and Schlemm's canal. CONCLUSIONS: These results indicate that at least a portion of aqueous outflow in the mouse eye is through the uveoscleral outflow pathway. PMID- 12091418 TI - Morphology of the murine optic nerve. AB - PURPOSE: To study the morphology of the murine optic nerve (ON). METHODS: Eyes of C57/Bl6 and BalbC mice were studied by light and electron microscopy. Microvascular castings of the ON region were prepared by transcardial injection of liquid plastic and studied with a scanning electron microscope. Immunohistochemistry was performed using antibodies against glial fibrillary acidic protein (GFAP), connexin 43, carbonic anhydrase II, and collagen types I and III. RESULTS: The transition between nonmyelinated and myelinated portion of the ON started approximately 0.6 mm behind the globe. A lamina cribrosa was completely absent. Instead, ON axons passed through a scleral hole that was surrounded by a ring of type III and type I collagen fibers. Instead of connective tissue beams within the nerve, layers of elongated astrocytes traversed the ON. All astrocytes stained for GFAP, but not for carbonic anhydrase II. The arterial supply of the nonmyelinated ON derived from branches of the central retinal artery. None of the capillaries derived from choroidal vessels. CONCLUSIONS: The mouse ON head differs from that of other species, because it lacks a lamina cribrosa and a choroidal vascular supply. Studies in glaucomatous mice might help to identify the importance of the lamina cribrosa and the choroidal vascular supply for optic nerve damage in glaucoma. PMID- 12091419 TI - Relationship between electrophysiological, psychophysical, and anatomical measurements in glaucoma. AB - PURPOSE: To evaluate the relationship between electrophysiological, psychophysical, and structural measurements in normal and glaucomatous eyes and to test the hypothesis that there is a continuous structure-function relationship between ganglion cell numbers and visual field sensitivity. METHODS: Thirty-four normal subjects and 40 patients with glaucoma were examined with the pattern electroretinogram (PERG), perimetry and retinal tomography. Transient and steady state (SS) PERGs were recorded, and peak (P)-to-trough (N) amplitude was measured. The unit of differential light sensitivity (DLS) in perimetry is the decibel. The decibel is 10. log(1/Lambert), where the Lambert is the unit of test spot intensity. PERG amplitudes were correlated with decibel and 1/Lambert DLS for the central 18 degrees of the visual field and with neuroretinal rim area in the temporal part of the optic disc. Age-related changes in the structural and functional measurements were sought. The correlation between variables was investigated by linear and quadratic regression analysis. A quadratic (y = ax + bx(2) + c) fit was taken to be significantly better than a linear fit, if the coefficient (b) for the x(2) term was significant at P < 0.05. RESULTS: A quadratic fit between decibel DLS and PERG amplitude (transient PERG: R(2) = 0.40, P = 0.0000; SS PERG: R(2) = 0.32, P = 0.0000) was significantly better than a linear fit. There was a linear correlation between 1/Lambert DLS and PERG amplitude (transient PERG: R(2) = 0.44, P = 0.0000; SS PERG: R(2) = 0.35, P = 0.0000). There was a linear correlation between temporal neuroretinal rim area and PERG amplitude (transient PERG: R(2) = 0.17, P = 0.0003; SS PERG: R(2) = 0.20, P = 0.0001). A quadratic fit between decibel DLS and temporal neuroretinal rim area (R(2) = 0.38, P = 0.0000) was significantly better than a linear fit. There was a linear correlation between 1/Lambert DLS and temporal neuroretinal rim area (R(2) = 0.30, P = 0.0000). Both DLS and PERG amplitude declined with age in the normal subjects. The rate of decline was -0.17%, -0.74%, -0.75%, and 0.78% per year for decibel DLS, 1/Lambert DLS, transient PERG, and SS PERG, respectively. CONCLUSIONS: There is a curvilinear relationship between decibel DLS and both PERG amplitude and neuroretinal rim area, and a linear relationship between 1/Lambert DLS and PERG amplitude and neuroretinal rim area. These findings support the hypothesis that there is no ganglion cell functional reserve but a continuous structure-function relationship, and that the impression of a functional reserve results from the logarithmic (decibel) scaling of the visual field. PMID- 12091420 TI - Individualized compensation of anterior segment birefringence during scanning laser polarimetry. AB - PURPOSE: To describe a method for assessment and individualized compensation of anterior segment birefringence with scanning laser polarimetry. METHODS: A scanning laser polarimeter (GDx Nerve Fiber Analyzer; Laser Diagnostic Technologies, Inc., San Diego, CA) was modified to accommodate a variable compensator. The magnitude and axis of anterior segment birefringence of normal eyes were determined from a polarimetry image of the Henle fiber layer. The variable compensator was then adjusted to minimize anterior segment birefringence. Retinal nerve fiber layer (RNFL) and macular measurements were then obtained. Macular images with individualized compensation served to verify the effectiveness of the compensation. To demonstrate individualized compensation, two sets of three images each were obtained from four eyes of four normal subjects. One set was obtained with individualized compensation and another with fixed compensation, as used in the commercial polarimetry system. RESULTS: In the tested eyes, the magnitude of anterior segment birefringence ranged from 21.7 to 86.3 nm, and the slow axis ranged from 5.7 degrees nasally upward to 54.3 degrees nasally downward. The maximum residual retardation resulting from compensation was 70 nm for fixed compensation and 11.5 nm for individualized compensation. The compensation residual directly affected the assessment of the RNFL by scanning laser polarimetry. RNFL images obtained with individualized compensation were more consistent with the expected anatomy of the eye. In the eyes measured, the range of RNFL thicknesses appeared to be narrower with the variable corneal and lens compensator (VCC) compared with the fixed corneal compensator (FCC). CONCLUSIONS: In eyes with a normal macula, the magnitude and axis of anterior segment birefringence can be determined from a polarimetry image of the Henle fiber layer. Individualized anterior segment compensation can be achieved with the described method so that the measured birefringence largely reflects the RNFL birefringence. Whether and how macular diseases affect this method remain to be investigated. PMID- 12091421 TI - Peripapillary scleral thickness in perfusion-fixed normal monkey eyes. AB - PURPOSE: To characterize the thickness of the peripapillary sclera in perfusion fixed normal monkey eyes so as to build accurate computational models of intraocular pressure (IOP)-related stress and strain within these tissues. METHODS: Nine rhesus monkeys were perfusion fixed, each with one normal eye set to an IOP of 10 mm Hg by manometer. A 6-mm-diameter specimen containing the optic nerve head and peripapillary sclera was trephined from each scleral shell and cut into 4-microm serial sagittal sections across the scleral canal opening, either horizontally (four eyes) or vertically (five eyes). The thickness of the peripapillary sclera was measured on every 24th section at 100-microm intervals from the posterior scleral canal opening (PSCO) to the peripheral edge of the specimen. The data were pooled by quadrant (superior, inferior, nasal, and temporal), regions within each quadrant, and distance from the PSCO, overall and for individual eyes, and subjected to analysis of variance. RESULTS: In terms of distance from the PSCO, the peripapillary sclera was thinnest nearest the PSCO (201 microm, nasal; 201 microm, temporal; 240 microm, inferior; 249 microm, superior), thickened progressively to a maximum in the midperiphery approximately 600 to 1000 microm from the PSCO (326 microm, nasal; 415 microm, superior; 420 microm, temporal; 422 microm, inferior), and thinned again peripherally in all quadrants. The peripapillary sclera was thinner in the nasal quadrant when compared with the other quadrants superiorly, inferiorly, and temporally (central region means of 291 microm, nasal; 369 microm, superior; 372 microm, inferior; and 369 microm, temporal; P < 0.0001). CONCLUSIONS: In the normal monkey eye, peripapillary scleral thickness varies significantly with distance from the posterior scleral canal opening and is thinner in the nasal quadrant than in the other quadrants. These differences are substantial and are likely to affect the magnitude of IOP-related stress and strain within these tissues for a given level of IOP. PMID- 12091422 TI - Retinal glutamate transporter changes in experimental glaucoma and after optic nerve transection in the rat. AB - PURPOSE: High levels of glutamate can be toxic to retinal ganglion cells. Effective buffering of extracellular glutamate by retinal glutamate transporters is therefore important. This study was conducted to investigate whether glutamate transporter changes occur with two models of optic nerve injury in the rat. METHODS: Glaucoma was induced in one eye of 35 adult Wistar rats by translimbal diode laser treatment to the trabecular meshwork. Twenty-five more rats underwent unilateral optic nerve transection. Two glutamate transporters, GLAST (EAAT-1) and GLT-1 (EAAT-2), were studied by immunohistochemistry and quantitative Western blot analysis. Treated and control eyes were compared 3 days and 1, 4, and 6 weeks after injury. Optic nerve damage was assessed semiquantitatively in epoxy embedded optic nerve cross sections. RESULTS: Trabecular laser treatment resulted in moderate intraocular pressure (IOP) elevation in all animals. After 1 to 6 weeks of experimental glaucoma, all treated eyes had significant optic nerve damage. Glutamate transporter changes were not detected by immunohistochemistry. Western blot analysis demonstrated significantly reduced GLT-1 in glaucomatous eyes compared with control eyes at 3 days (29.3% +/- 6.7%, P = 0.01), 1 week (55.5% +/- 13.6%, P = 0.02), 4 weeks (27.2% +/- 10.1%, P = 0.05), and 6 weeks (38.1% +/- 7.9%, P = 0.01; mean reduction +/- SEM, paired t-tests, n = 5 animals per group, four duplicate Western blot analyses per eye). The magnitude of the reduction in GLT-1 correlated significantly with mean IOP in the glaucomatous eye (r(2) = 0.31, P = 0.01, linear regression). GLAST was significantly reduced (33.8% +/- 8.1%, mean +/- SEM) after 4 weeks of elevated IOP (P = 0.01, paired t test, n = 5 animals per group). In contrast to glaucoma, optic nerve transection resulted in an increase in GLT-1 compared with the control eye (P = 0.01, paired t-test, n = 15 animals). There was no significant change in GLAST after transection. CONCLUSIONS: GLT-1 and GLAST were significantly reduced in an experimental rat glaucoma model, a response that was not found after optic nerve transection. Reductions in GLT-1 and GLAST may increase the potential for glutamate-induced injury to RGC in glaucoma. PMID- 12091423 TI - Detection of herpes simplex virus type 1 in human ciliary ganglia. AB - PURPOSE: To determine whether herpes simplex virus type 1 (HSV-1) DNA is present in the ciliary ganglion (CG). METHODS: Fifty CG and 47 trigeminal ganglia (TG) were resected from 63 formalin-fixed cadavers between 56 and 98 years of age that had been embalmed within 12 hours of death. The donors had no known active HSV infection at the time of death. DNA was extracted from all ganglia by proteinase K digestion (TG) or digestion by a mild lysis buffer (CG). DNA was amplified by polymerase chain reaction for sequences from human chromosome 18, D18S1259 (positive control), and from the HSV-1 DNA polymerase gene, U(L)30. The amplified DNA was separated by agarose gel electrophoresis, transferred to nylon membranes, and hybridized with the appropriate digoxigenin-labeled probe that was detected by alkaline phosphatase-conjugated monoclonal antibody. RESULTS: The D18S1259 sequence was amplified from 47 TG and 30 CG samples. Of these samples, 32 (68.0%) of the 47 TG samples and 20 (66.6%) of the 30 CG samples were positive for the UL(30) HSV-1 sequence. CONCLUSIONS: Using amplification of HSV-1 DNA as a surrogate marker of latency, the finding that the frequency of HSV-1 in the CG was approximately the same as that of the TG suggests that the CG may be an additional site of HSV-1 latency in humans. Active infection in or reactivation of HSV-1 from non-TG sites may explain why this virus is able to infect sites, such as the retina, that have no direct connections to the trigeminal nerve. PMID- 12091424 TI - Retinal microenvironment controls resident and infiltrating macrophage function during uveoretinitis. AB - PURPOSE: Macrophages infiltrating an inflamed or injured tissue undergo development of coordinated sets of properties that contribute to tissue damage, repair, and remodeling. The purpose of this study was to determine whether macrophages isolated from normal or inflamed retina are programmed to a distinct set of properties and to examine whether the development of experimental autoimmune uveoretinitis (EAU) affects macrophage function. METHODS: EAU was induced in Lewis rats, and a retina-derived macrophage-enriched population was generated by density centrifugation during the prepeak, peak, and resolution phases of the disease. Cell surface phenotype was assessed by two- and three color flow cytometry, and function was determined in vitro by nitric oxide (NO) production, with or without further cytokine stimulation or by immunohistochemistry to determine expression of beta-glucuronidase, nitric oxide synthase (NOS)-2, and nitrotyrosine. RESULTS: Myeloid-derived cells from normal retina were programmed similar to TGF-beta-stimulated uncommitted bone-marrow derived macrophages (BMDMs). Contrary to BMDM behavior, retina-isolated macrophages displayed distinct properties and phenotype at different phases of the disease course and remained resistant throughout, to further cytokine challenge in vitro. During peak disease, retina-isolated macrophages had characteristics of IFN-gamma/TNF-alpha primed cells (nitrotyrosine positive and NO producing). Despite equivalent numbers of macrophages during resolution, their function reverted to characteristics of TGF-beta primed cells (beta-glucuronidase positive). CONCLUSIONS: Resident retinal myeloid-derived cells are primed and are resistant to further cytokine stimulation, and, similar to macrophages derived during EAU recovery, behave operationally as though TGF-beta primed. During peak inflammation, infiltrating macrophages adapt to concurrent hierarchical Th1 T cell response (IFN-gamma/TNF-alpha), generating NO. The results provide evidence of in vivo programming of macrophages within the retina. PMID- 12091425 TI - RPE65 is highly uveitogenic in rats. AB - PURPOSE: To examine the hypothesis that RPE65, a protein specific to the retinal pigment epithelium, is uveitogenic in rats. METHODS: Rats of four inbred strains (Lewis, Brown Norway, Fischer, and SHR) were immunized with native or recombinant bovine RPE65, or with S-antigen (S-Ag), emulsified with complete Freund adjuvant, and treated simultaneously with killed Bordetella pertussis bacteria, as indicated. Development of ocular changes was examined and scored both clinically and histologically. RESULTS: Lewis rats immunized with RPE65 showed development of acute and severe inflammatory eye disease that affected most ocular tissues. The minimum uveitogenic dose of RPE65 was similar to that of S-Ag (1 microg per rat), but the changes induced by RPE65 at higher dose ranges were less severe than those induced by S-Ag. Concurrent treatment of the RPE65-immunized rats with B. pertussis bacteria was not critical for disease induction, but enhanced dramatically the pathogenic reaction. Unlike the results with several other retinal proteins, no pinealitis was detected in rats immunized with RPE65. Fischer (F344) rats resembled Lewis rats in being similarly affected by RPE65 or S-Ag. In contrast, Brown Norway (BN) rats developed severe disease when immunized with RPE65, but showed minimal changes in response to S-Ag. SHR rats responded poorly to disease induced by RPE65, and S-Ag-induced disease failed to develop. CONCLUSIONS: RPE65 is highly uveitogenic in rats, thus suggesting that this molecule could be involved in pathogenic autoimmunity in the human eye. PMID- 12091427 TI - Lipoteichoic acid selectively induces the ERK signaling pathway in the cornea. AB - PURPOSE: To identify signal transduction pathways and gene expression induced by the bacterial cell wall component lipoteichoic acid (LTA) in human corneal keratocytes. METHODS: Human corneal keratocytes were cultured in the presence of 6.25 to 50 microg/ml LTA from Staphylococcus aureus. Induced DNA-binding of NF kappaB was determined by electrophoretic mobility shift assays (EMSAs). Activation of MAP-kinase signaling pathways (p38, JNK-1/2, ERK-1/2, Elk 1, MEK 1/2, c-Raf) was evaluated by Western blotting using phospho-specific antibodies. To investigate the effect of LTA signaling on gene expression, keratocytes were transfected with a luciferase reporter gene under the control of serum response elements (SREs). LTA-induced gene expression was quantified using luciferase assays. RESULTS: Exposure of various concentrations of LTA up to 24 hours did not lead to activation of NF-kappaB, whereas TNF-alpha potently induced this transcription factor. A systematic analysis of LTA-activated MAPK pathways revealed no significant effects on JNK and p38, but a dose- and time-dependent phosphorylation of members of the ERK pathway. Analysis of the ERK-activating signaling cascade showed LTA-induced phosphorylation of ERK-1, MEK1/2, and c-Raf. ERK activity resulted in an enhanced transcription of an SRE-controlled reporter gene. CONCLUSIONS: LTA induces SRE-enhanced gene transcription in corneal keratocytes that is selectively mediated by the ERK pathway. Therefore, it seems possible that components of the bacterial cell wall such as LTA can alter the transcriptional program within the corneal stroma and thereby trigger an inflammatory response. PMID- 12091428 TI - Regulation of endotoxin-induced keratitis by PECAM-1, MIP-2, and toll-like receptor 4. AB - PURPOSE: Bacterial lipopolysaccharide (LPS, endotoxin) is a potent stimulator of inflammatory responses and is likely to contribute to microbial keratitis and to the pathogenesis of sterile corneal ulcers. The purpose of the present study was to identify specific mediators of endotoxin-induced keratitis. METHODS: The corneal epithelium of BALB/c, C3H/HeJ, and C3H/HeN mice was abraded, and Pseudomonas aeruginosa endotoxin (10 microg in 1 microL) was added. Stromal thickness and haze were measured by in vivo scanning confocal microscopy, and neutrophil recruitment determined by immunohistochemistry. RESULTS: Pseudomonas endotoxin induced a significant increase in stromal thickness and haze compared with untreated control corneas at each time point examined, and the severity coincided with neutrophil infiltration into the corneal stroma. Furthermore, systemic depletion of neutrophils completely abrogated endotoxin-induced increases in stromal thickness and haze, indicating an essential role for these cells. Expression of platelet endothelial cell adhesion molecule (PECAM)-1 on vascular endothelium and production of macrophage inflammatory protein (MIP)-2 in the corneal stroma were also significantly elevated after exposure to endotoxin, and antibody blockade inhibited neutrophil recruitment to the cornea and abrogated endotoxin-induced increases in stromal thickness and haze. In LPS hyporesponsive C3H/HeJ mice, PECAM-1 and MIP-2 were not upregulated after exposure to endotoxin, and endotoxin-induced keratitis did not develop in these mice. CONCLUSIONS: These findings demonstrate that endotoxin-induced keratitis is regulated by toll-like receptor-4 (TLR4)-dependent expression of PECAM-1 and MIP 2, which are essential for recruitment of neutrophils to this site and for development of endotoxin-induced stromal disease. PMID- 12091426 TI - Identification of a novel macrophage population in the normal mouse corneal stroma. AB - PURPOSE: To examine the normal murine corneal stroma for the presence of bone marrow-derived leukocytes. METHODS: Wholemounts of paraformaldehyde-fixed corneal stroma from normal mice at 5 to 16 weeks of age were examined in single- and double-color immunomorphologic studies performed with confocal microscopy. The phenotype, morphology, distribution, and density of immunopositive cells were determined. RESULTS: Numerous CD45(+) cells with pleomorphic and dendriform morphology were found within the pericentral and central region of the corneal stroma (200-300 cells/mm(2)). Dual-color immunostaining demonstrated that 100% of the CD45(+) cells coexpressed CD11b and 50% coexpressed F4/80. Approximately 30% of the total cells and 50% of the F4/80(+) cells coexpressed major histocompatibility complex (MHC) class II antigens. Very small to negligible numbers of cells expressed markers of dendritic cells (CD11c) or granulocytes (Ly6G). Markers for T-cells and NK cells were absent from the corneal stroma, indicating that all the cells identified in the stroma were of the myeloid lineage. CONCLUSIONS: The normal murine corneal stroma contains a significant number of CD45(+) leukocytes. Most these cells express the CD11b marker, but not other dendrite, granulocyte, T-cell, or NK markers, placing them in the monocyte/macrophage lineage. PMID- 12091429 TI - Inhibition of fiber cell globulization and hyperglycemia-induced lens opacification by aminopeptidase inhibitor bestatin. AB - PURPOSE: To examine the role of calcium-dependent and -independent proteolytic activity in the globulization of isolated fiber cells and glucose-induced lens opacification. METHODS: Fiber cells from rat lens cortex were isolated, and the [Ca(2+)](i) and protease activity in the isolated fibers were determined by using a calcium binding dye and the protease substrate t-butoxycarbonyl-Leu-Met-7-amino 4-chloromethylcoumarin (BOC-Leu-Met-CMAC). The activity of calpain in the lens cortex homogenate was determined with fluorescein-casein in the presence of Ca(2+) and that of fiber cell globulizing aminopeptidase (FCGAP) with BOC-Leu-Met CMAC and reduced glutathione (GSH) in the absence of Ca(2+). The lens proteases calpain and the novel aminopeptidase FCGAP were partially purified by diethylaminoethyl (DEAE) gel column chromatography. Single fiber cells were isolated from rat lens, plated on coverslips, and placed in a temperature controlled chamber. Their globulization time was determined by the appearance of light-scattering globules in the absence and the presence of protease inhibitors including the aminopeptidase inhibitor bestatin. To investigate the effect of the protease inhibitors E-64 and bestatin on the prevention of hyperglycemic cataract, the rat lenses were cultured in medium 199 in the presence of 5.5 and 50 mM glucose and in the absence and the presence of protease inhibitors. Changes in light transmission by the lenses were determined by digital image analysis. RESULTS: Normal levels of lens fiber cell [Ca(2+)](i), determined by using a cell permeable dye were approximately 100 nM, and the protease activity determined with BOC-Leu-Met-CMAC was maximum at [Ca(2+)](i) of approximately 500 nM. A large fraction of the FCGAP that cleaves BOC-Leu-Met-CMAC was separated from calpain, which cleaves fluorescein-casein, by diethylaminoethyl (DEAE) gel column chromatography. The FCGAP did not bind to the column, whereas calpain bound to the column and was eluted by approximately 180 mM NaCl. Unlike calpain, the FCGAP did not require calcium for activation and did not cleave fluorescein-casein. However, the Ca(2+)-dependent calpain activated FCGAP, indicating that the latter may exist in pro-protease form. The FCGAP was selectively inhibited by the specific aminopeptidase inhibitor bestatin, indicating that FCGAP could be an aminopeptidase. However, the FCGAP was found to be immunologically distinct from leucine aminopeptidase and calpain. Perfusion of the isolated rat lens fiber cells with Ringer's solution led to their globulization in 30 +/- 3 minutes. Addition of 0.5 mM of the protease inhibitors E-64 and leupeptin increased the globulization time to 60 and 100 minutes, respectively, whereas no globulization of the fiber cells was observed for 4 hours in the presence of 0.05 mM bestatin. In rat lens cultured in medium containing 50 mM glucose, both E-64 and bestatin (0.05 mM each) significantly reduced the extent of opacification, indicating that an aminopeptidase, downstream to a Ca(2+)-dependent protease, may be involved in mediating cataractogenic changes. CONCLUSIONS: In addition to calpain, a Ca(2+) independent novel protease, FCGAP, a novel aminopeptidase, represents a significant fraction of the total proteolytic activity in the lens. Inhibition of FCGAP by bestatin attenuates Ca(2+)-induced globulization of the isolated fiber cells in vitro and hyperglycemia-induced opacification of cultured rat lens. PMID- 12091430 TI - The role of Src family kinases in cortical cataract formation. AB - PURPOSE: The goal of this study was to determine the role of Src family kinases (SFKs) in the development of lens cataract. This question was particularly significant, because these tyrosine kinases mediate the stress pathways known to lead to cataract formation. The experiments were focused on whether the inhibition of SFK activity suppresses the formation of lens opacities. METHODS: A whole-lens culture system was developed, in which cortical opacities formed within 5 days, in embryonic day (E)10 lenses grown in medium containing 10% fetal bovine serum. SFK activity was blocked in the cultured lenses by growth in the presence of the SFK-specific inhibitor PP1. Control cultures were grown in medium without inhibitor or in the presence of PP3, the inactive analogue of PP1. Lenses were cultured for 10 days, observed, and photographed daily. Opacification was quantified with image-analysis software. Tissue architecture was determined after hematoxylin and eosin staining and cellular organization by fluorescent localization of filamentous actin with fluorescein-conjugated phalloidin. RESULTS: Almost all lenses in the control cultures developed cortical opacities covering approximately 50% of the lens area by day 10. Similar to control cultures, PP1-treated lenses showed mild posterior opacities during the first 5 days in culture, but then became strikingly transparent. Only 7% of the PP1 treated lenses showed development of cortical cataract, and the average area of opacity was just 0.5% by culture day 10. In all cultured lenses, even in the presence of the PP1 inhibitor, the bow region of the lens extended to the posterior pole, and distribution of nuclei from the posterior pole toward the anterior aspects of the lens suggested that newly added fiber cells were misdirected. However, neither this feature, nor the presence of vacuoles appeared to correlate with the development of opacity in the cultured lenses. Instead, the lens opacities appeared to result from gross abnormalities in the shape and organization of cells in the equatorial and cortical fiber zones, as observed by F-actin staining. Culturing the lenses in the presence of the SFK inhibitor prevented these lens cell aberrations as well as the development of lens opacity. CONCLUSIONS: The formation of cataract can involve activation of SFK-mediated pathway(s) leading to disorganization of developing lens fiber cells, and inhibiting these tyrosine kinases blocks cataract progression. PMID- 12091432 TI - Flicker-evoked changes in human optic nerve blood flow: relationship with retinal neural activity. AB - PURPOSE: Visual flicker induces a response in human optic nerve blood flow (F(onh)) and inner retinal activity, as assessed by laser Doppler flowmetry and electroretinogram (ERG), respectively. In this study the relationship was examined between the flicker-evoked changes in F(onh) (RF(onh)) and ERG when various parameters of the stimulus were varied. METHODS: In five normal observers (mean age, 41; range, 25-62 years) F(onh) and ERG were recorded simultaneously in response to pure red (R) or pure green (G) flicker, as well as heterochromatic R G flicker (30 degrees field at the posterior pole). RF(onh) and the changes in the first (1F) and second (2F) harmonic amplitudes of the ERG were documented as a function of the frequency of pure luminance and equiluminant R-G flicker, the mean illuminance of 10-Hz luminance flicker, and the color ratio r = R/(R + G) of a 15-Hz heterochromatic R-G flicker. RESULTS: Frequency-dependent changes in RF(onh) were similar to those in both 1F and 2F amplitudes for equiluminant R-G flicker. RF(onh) and 2F amplitude increased and then saturated with increasing mean illuminance of luminance flicker. They both decreased similarly as the R-G flicker approached the r value corresponding to equiluminance. RF(onh) was positively correlated with both 1F and 2F amplitudes (r = 0.55 and 0.31, respectively, P < 0.05) when these quantities were recorded as a function of frequency of R-G equiluminant flicker. RF(onh) was positively correlated with 2F amplitudes when both quantities were recorded as a function of mean illuminance of luminance flicker and r of heterochromatic R-G flicker (r = 0.52 and 0.48, respectively, P < or = 0.01). CONCLUSIONS: Under specific experimental conditions, changes in human RF(onh) are similar to and correlated with those of the flicker ERG 1F and 2F amplitudes. These findings support a relationship between vaso- and neural activity changes in the neural tissue of the human eye. PMID- 12091431 TI - TGF-beta2-induced matrix modification and cell transdifferentiation in the human lens capsular bag. AB - PURPOSE: To study the role of TGF-beta2 in posterior capsule opacification (PCO) and to determine whether CAT-152 (lerdelimumab), a fully human monoclonal antibody that neutralizes the effect of TGF-beta2, can also provide therapeutic benefit for PCO. METHODS: In vitro capsular bags were prepared from human donor eyes and maintained in a 5% CO(2) atmosphere at 35 degrees C. To investigate expression of active TGF-beta2, capsular bags were incubated in serum-free EMEM for 2, 28, or more than 100 days and analyzed by ELISA (n > or = 4 at each time point). To study underlying mechanisms, match-pair experiments were also performed, so that the medium was supplemented with 0, 1 or 10 ng/mL TGF-beta2 with or without 10 microg/mL CAT-152 (n = 4 in all cases). On-going observations were by phase-contrast microscopy. In addition, donor material from patients who had undergone cataract surgery was analyzed. Cellular architecture was examined by fluorescence cytochemistry. Expression of matrix metalloproteinase (MMP)-2 and -9 was assessed by gelatin zymography. RESULTS: Analysis of capsular bags from donor eyes that had received an intraocular lens (IOL) revealed the presence of endogenous active TGF-beta2, matrix wrinkling, and expression of transdifferentiation markers alphaSMA and fibronectin. When cultured in vitro, donor bags also showed sustained release of MMP-2 and -9. Culture of capsular bags prepared in vitro from whole lenses showed that TGF-beta2 (1-10 ng/mL) stimulated transdifferentiation and contraction of the capsular bag, resulting in light scatter. TGF-beta2 also induced sustained release of MMP-2 and -9. Active TGF-beta2 was detected in these cultures. The human monoclonal anti-TGF-beta2 antibody CAT-152 (10 microg/mL) effectively inhibited all TGF-beta2-induced effects. CONCLUSIONS: Addition of TGF-beta2 accelerates transdifferentiation and contraction of the capsular bag, resulting in light scatter. CAT-152 inhibited all the effects of TGF-beta2 that were examined and therefore has the potential to suppress development of PCO and provide potential therapeutic benefit to cataract patients. PMID- 12091433 TI - P2 purinergic receptor-coupled signaling in the rabbit ciliary body epithelium. AB - PURPOSE: To identify and characterize P2 purinergic receptors and their signaling pathways in the epithelial cells of the rabbit ciliary body. METHODS: Real-time fluorescence ratio imaging of the intact fura-2-loaded nonpigmented ciliary body epithelial (NPE) cells of rabbit were used to record changes in the intracellular free calcium concentration ([Ca(2+)](i)), in response to a number of purinergic agonists and antagonists. The effects of some of these drugs on the inositol phosphate (IP) levels in ciliary processes were also examined. RESULTS: Adenosine diphosphate (ADP), adenosine triphosphate (ATP), and uridine triphosphate (UTP) dose dependently increased the [Ca(2+)](i) and IP levels. The [Ca(2+)](i) increases induced by ADP and UTP were distinguishable, both kinetically and pharmacologically. The effect of ADP on [Ca(2+)](i) was mimicked by a number of P2Y(1)-selective agonists, and was blocked by three P2Y(1)-receptor-specific antagonists. The [Ca(2+)](i) increases elicited by ADP (or its analogs) and UTP were additive. CONCLUSIONS: Rabbit ciliary body epithelium possesses both P2Y(1) and P2Y(2) metabotropic purinergic receptor subtypes, which differentially use the IP(3)/Ca(2+) second-messenger pathway. PMID- 12091434 TI - Interaction of GABA receptor/channel rho(1) and gamma(2) subunit. AB - PURPOSE: To determine whether protein-protein and functional interactions can occur between gamma-aminobutyric acid (GABA)(A) receptor/channels gamma(2) subunit and the retina-specific GABA(C) rho(1) subunit. METHODS: Protein-protein interaction was characterized by immunocoprecipitation of these subunits in brain and spinal cord with anti-gamma(2) subunit antibody and by Western blot analysis with anti-rho(1) subunit antibody. The rho(1) and gamma(2) subunits were detected in the adult rat brain and spinal cord lysates that had been previously precipitated with the specific antibodies against the rho(1) and gamma(2) subunits, respectively. A two-microelectrode voltage clamp was used to measure GABA-induced currents in oocytes. In addition, a yeast two-hybrid system was used to detect the interactions of these subunits in vivo. RESULTS: Based on yeast transformed with the N-terminal fragment of the gamma(2) subunit (gamma(2)-N'), the N-terminal fragment of the rho(1) subunit (rho(1)-N'), and the full-length rho(1) subunit, the protein-protein interaction of the GABA(A) gamma(2) subunit and the GABA(C) rho(1) subunit was found in yeast grown in triple-dropout medium (deficient in Leu, Trp, and His) and expressing the LacZ reporter gene. Interaction of the rho(1) and gamma(2) subunits was investigated by functional studies in which gamma(2) (gamma(2)-N' with 837 bp) and rho(1) cRNAs were coinjected in Xenopus oocytes. In studies of the functional interaction, after injection of the gamma(2) subunit mutant cRNA containing a N-terminal fragment, GABA-induced rho(1) originated currents declined to 16% of the control level of homooligomeric rho(1) current. This inhibitory effect of coexpressing gamma(2) subunit mutants with rho(1) subunit on the rho(1)-originated current in oocytes was dose dependent. In addition, coexpression of the GABA rho(1) and gamma(2) subunits in oocytes altered pharmacologic properties of the homooligomeric receptor/channel formed by rho(1) or gamma(2) subunits. Further evidence was provided by results obtained with specific antibodies showing that the rho(1) subunit was coimmunoprecipitated with the gamma(2) subunit from the retina, brain, and spinal cord. CONCLUSIONS: The results indicate that protein-protein and functional interactions can occur between the GABA(A) gamma(2) subunit and the GABA(C) rho(1) subunit. Therefore, the functional role of GABA receptor/channels in the brain, retina, and spinal cord is more diversified because of the possible assembly between the GABA(A) gamma(2) subunit and GABA(C) rho(1) subunit. PMID- 12091435 TI - Intraocular production and release of nerve growth factor after iridectomy. AB - PURPOSE: To determine the presence of nerve growth factor (NGF), NGF mRNA, and NGF receptor (TrkA) in rabbit ocular tissues, and whether changes occur in NGF and NGF mRNA levels after experimental iridectomy. METHODS: Immunohistochemistry for NGF and TrkA and in situ hybridization for NGF mRNA were performed on rabbit cornea, iris, ciliary body, and lens in the basal state. Quantification of NGF mRNA and NGF protein levels in these tissues was performed by RT-PCR and immunoenzymatic assay, respectively. A time course of NGF concentration in the aqueous humor and the expression of NGF mRNA in iris and ciliary body were performed after the iridectomy and were compared with levels in a sham-treated group (paracentesis). RESULTS: Cornea, iris, ciliary body, and lens expressed NGF mRNA, NGF protein, and TrkA in the basal state. The highest levels of NGF were detected in the iris (8938.0 +/- 3968.1 pg/g), and the lowest were in the aqueous humor (22.8 +/- 9.7 pg/mL). Experimental iridectomy induced a transient increase of NGF concentration in the aqueous humor that reached its peak 4 hours after the experimental injury (464.4 +/- 29.9 pg/mL versus the control group 101.6 +/- 18.8 pg/mL; P < 0.001) and returned to baseline value after 7 days. A significant increase of NGF mRNA was also observed 1 hour and 4 hours after the iridectomy in the iris (1 hour, 788 +/- 85 OD; 4 hours, 760 +/- 81 OD versus baseline, 246 +/- 32 OD; P < 0.0001) and ciliary body (1 hour, 330 +/- 19 OD; 4 hours, 453 +/- 52 OD versus baseline, 219 +/- 37 OD; P < 0.05), but not in the cornea, lens, or any tissues from the control group. CONCLUSIONS: NGF is present and produced in the anterior segment of the eye and is released in the aqueous humor in the basal state. Experimental iridectomy induces increased production of NGF in the iris and in the ciliary body and an increased concentration of NGF in the aqueous humor. PMID- 12091436 TI - Lysosomal Ca(2+) stores in bovine corneal endothelium. AB - PURPOSE: Acidic organelles, including Golgi bodies and lysosomes, are known to operate as Ca(2+) storage sites in many cell types. This study demonstrates the presence of Ca(2+) stores in lysosomes of bovine corneal endothelial cells (BCECs) and examines their interaction with Ins(1,4,5)P(3)-sensitive Ca(2+) stores. METHODS: Glycyl-L-phenylalanine-beta-naphthylamide (GPN) was used to release Ca(2+) from lysosomes by inducing their selective osmotic swelling. Ca(2+) released into the cytoplasm was measured with fura-2 or fura-PE3 fluorescent dyes. Fluorescence of acridine orange (AO), which selectively sequesters into acidic organelles, was used to establish swelling of lysosomes in response to GPN. RESULTS: Exposure to GPN (100-200 microM) in cultured BCECs produced an increase in free cytosolic Ca(2+) ([Ca(2+)](i)) equivalent to approximately 79% of the peak response to uridine triphosphate (UTP), a P2Y agonist (n = 19). The endothelium of the freshly isolated cornea also produced [Ca(2+)](i) transients similar to those in cultured BCECs; however, the peak [Ca(2+)](i) increase was smaller ( approximately 43% of the peak response to UTP; n = 13). In cultured BCECs, the response to UTP was unaffected by pretreatment with GPN with extracellular calcium ([Ca(2+)](o)) at 0 and 1.2 mM (n = 10). Neither pretreatment with thapsigargin (5 microM) nor with U73122 (a phospholipase C inhibitor; 10 microM) blocked the peak GPN response (n = 6). Exposure to 20 microM monensin produced a [Ca(2+)](i) increase with [Ca(2+)](o) at 0 and 1.2 mM and also reduced the subsequent peak response to GPN (n = 6). CONCLUSIONS: GPN-sensitive lysosomal Ca(2+) stores, distinct from Ins(1,4,5)P(3) sensitive Ca(2+) stores, are found in both cultured cells and fresh tissue. These stores are susceptible to depletion by the loss of the pH gradient across lysosomes and P2 agonists. The latter occurs through mechanisms independent of phospholipase C (PLC) activation or Ins(1,4,5)P(3). The GPN stores also induce [Ca(2+)](o) influx in response to their depletion. PMID- 12091437 TI - Twenty-four-hour pattern of intraocular pressure in young adults with moderate to severe myopia. AB - PURPOSE: To characterize the 24-hour change of intraocular pressure (IOP) in young adults with moderate to severe myopia. METHODS: Nineteen young adults, ages 18 to 25 years, with moderate to severe myopia (myopia group) and 17 age-matched volunteers with emmetropia or mild myopia (control group) were housed for 1 day in a sleep laboratory. An 8-hour accustomed sleep period was assigned to each volunteer. Twelve measurements of IOP, axial length, blood pressure, and heart rate were taken at 2-hour intervals. In the wake period, blood pressure and heart rate were measured after a 5-minute bed rest. Axial length and IOP were measured in supine volunteers. Volunteers then sat for 5 minutes, after which IOP was measured. In the sleep period, measurements were taken in supine volunteers in bed. RESULTS: In both the myopia and control groups, the average supine IOP in the sleep period was higher than the average sitting IOP in the wake period. However, the magnitude of this IOP elevation at night was significantly less in the myopia group. In the sleep period, IOP was less in the myopia group than in the control group. When only the 24-hour supine IOP data were considered, the trough occurred at 1:30 AM, and the peak occurred around noon in the myopia group. In the control group, the trough was at 9:30 PM, and the peak at 5:30 AM. Least-square cosine fits showed 24-hour rhythms of supine IOP in both groups, but their phase timings were different. Axial length remained unchanged throughout the day and night in both groups. There was no difference in the 24-hour rhythms of mean blood pressure and heart rate between the two groups. CONCLUSIONS: Considering habitual body positions, IOP increases at night in young adults with moderate to severe myopia, but the magnitude of the increase is significantly less than that in the age-matched control subjects. There is a 24-hour rhythm of supine IOP in the myopic group, but the phase timing is different from that in the control subjects. These variations of IOP in young adults with moderate to severe myopia are not related to changes in cardiovascular parameters. PMID- 12091438 TI - Glucose-induced activation of glucose uptake in cells from the inner and outer blood-retinal barrier. AB - PURPOSE: The purpose of this study was to elucidate in vitro the effect of elevated glucose on glucose uptake in the cells comprising the inner and outer blood-retinal barriers: human retinal pigment epithelial (hRPE) and human retinal vascular endothelial (hRVE) cells. METHODS: Primary cultures of hRPE and hRVE cells grown in 5.5 or 22 mM glucose or in 22 mM mannitol were used to measure the rates of glucose uptake with [(3)H]-3-O-methyl glucose as a tracer. GLUT1 expression was measured by Northern and western blot analyses. Cellular fractionation was performed by differential centrifugation. GLUT1 overexpression was accomplished by adenoviral transduction. RESULTS: Increasing media glucose from 5.5 to 22 mM for 30 minutes caused a 1.9-fold increase in the V(max) of glucose uptake in hRPE cells and a 2.5-fold increase in hRVE cells. These increases were nonosmotic and glucose specific, in that the exposure to 22 mM mannitol did not affect the V(max) of glucose uptake. mRNA, total protein expression, and translocation of GLUT1, the glucose transporter predominantly expressed in hRPE and hRVE cells, were not affected by 22 mM glucose for up to 48 hours. High-glucose effects on V(max) were abolished with 10 microg/mL of the microtubule assembly inhibitor nocodazole. hRPE cells transduced to overexpress GLUT1 showed a 1.5-fold increase in the V(max) for glucose uptake versus control transduced cells. However, the magnitude of glucose-induced increase in glucose uptake was the same in GLUT1- and control-transduced cells. CONCLUSIONS: High glucose induced 1.9- and 2.5-fold increases in the V(max) of glucose uptake in hRPE and hRVE cells, respectively. These increases were not due to an increase in GLUT1 expression. The increases were dependent on microtubule integrity, but not on GLUT1 translocation. The mechanism of the increases is unknown. GLUT1 regulating protein(s) and/or novel glucose transporter(s) may be involved in the regulation of glucose uptake by glucose in the cells that comprise the blood retinal barrier. PMID- 12091439 TI - Local cone and rod system function in progressive cone dystrophy. AB - PURPOSE: To compare the patterns of local cone and rod system impairment in patients with progressive cone dystrophy (CD) using psychophysical and electrophysiological techniques. METHODS: Local cone system function was assessed by measuring cone system thresholds (visual fields) and cone-mediated multifocal electroretinograms (mfERGs). Rod system function was assessed by measuring rod system thresholds (visual fields) and rod-mediated mfERGs. The results in a group of eight patients with CD were compared with those in an age-similar control group. RESULTS: All the patients had abnormal cone system visual field thresholds and cone-mediated mfERGs. Cone system psychophysical thresholds were elevated for targets presented within the central 10 degrees, but were within normal limits for targets at peripheral locations. Cone-mediated mfERG measures of amplitude scale and time scale were abnormal for most of the hexagons tested. Most of the rod-mediated psychophysical thresholds and mfERGs were within normal limits. Rod system losses tended to be patchy and scattered throughout the area tested. CONCLUSIONS: There was poor correspondence among local measures of cone and rod system losses in these patients with CD. The results suggest that the spatial pattern of cone system losses in this disease differs from the spatial pattern of rod system losses. PMID- 12091440 TI - The retina of Spalax ehrenbergi: novel histologic features supportive of a modified photosensory role. AB - PURPOSE: The retina of the blind mole rat Spalax ehrenbergi was compared with other vertebrate photosensitive organs in an attempt to correlate its histologic organization with a presumptive nonvisual photoreceptor role. METHODS: The eyes of eight adult animals were analyzed by light and electron microscopy, using conventional staining and immunolabeling with antibodies against phototransduction proteins and calretinin. RESULTS: Rods accounted for most of the photoreceptor cells in the Spalax retina, although their morphology is dissimilar to that of sighted mammals, in that they contained only rudimentary outer segments. The latter showed strong rod-opsin and transducin immunoreactions. The phagosomes in the retinal pigmentary epithelium were also rod-opsin positive. Synapses were evident at the photoreceptor cells pedicles. Occasionally, several synaptic active sites were present, suggesting cone cell origin; however, cone-opsin was not immunodetected in the study samples. Synaptic ribbon fields, sometimes distant to the active sites, resembled those found in the vertebrate pineal. The other retinal layers were somewhat less organized than in sighted mammals. Some cells were displaced and the calretinin-positive inner plexiform layer had no sublayers. Calretinin immunolabeling was found in horizontal, amacrine, and ganglion cells. Folding of the retina produced rosette like images similar to those reported before in the retina of nocturnal mammals and in the avian pineal gland. CONCLUSIONS: These data suggest that the retina of the mole rat has undergone evolutionary restructuring to a photoreceptive pineal like organization. This supports the thesis that the photoreceptor cells of this unique organ have been reprogrammed during the subterranean adaptation of Spalax, from their original visual function to mediating photoperiodic regulation. PMID- 12091441 TI - Verteporfin photodynamic therapy in the rat model of choroidal neovascularization: angiographic and histologic characterization. AB - PURPOSE: To develop a model of verteporfin photodynamic therapy (PDT) for experimental choroidal neovascularization CNV in the rat. METHODS: A laser injury model was used to induce experimental CNV in rats. The transit and accumulation of the photosensitizer verteporfin was assessed angiographically in CNV lesions, to determine the optimal time for delivery of light energy. The CNV lesions were then treated with verteporfin PDT, with two doses of verteporfin (3.0 and 6.0 mg/m(2)) and four activating doses of light energy (10, 25, 50, and 100 J/cm(2)). Closure of the CNV was assessed both angiographically and histologically. Verteporfin PDT was also performed on areas of normal choroid and retina at the two verteporfin doses and four light energy doses. The effect of these treatments on these structures was also assessed angiographically and histologically. RESULTS: Peak verteporfin intensities in the CNV were detected at 15 to 20 minutes after intravenous injection. Rates of closure of the CNV varied as a function of the dose of verteporfin and of the activating light energy. Angiographic closure of the CNV correlated with damage to the neovascular complex, as seen with light and electron microscopy. Damage to areas of normal choroid and retina treated with verteporfin PDT also varied as a function of the verteporfin and light energy doses. CONCLUSIONS: Verteporfin PDT for experimental CNV in the rat is a feasible, effective, and reproducible model that can be used for testing the efficacy of adjunctive therapy to verteporfin PDT. PMID- 12091442 TI - Change of redox status and modulation by thiol replenishment in retinal photooxidative damage. AB - PURPOSE: Cellular or tissue reduction-oxidation (redox) is crucial in various diseases. The present study was conducted to analyze how tissue redox status is affected by photooxidative stress and whether the exogenous thiol antioxidant N acetylcysteine (NAC) affects photooxidative stress-induced retinal damage. METHODS: Mice were intraperitoneally injected with either NAC (250 mg/kg) or phosphate-buffered saline (PBS) and exposed to white fluorescent light (8000 lux) for 2 hours. Levels of thioredoxin (TRX), glutaredoxin (GRX), and glutathione (GSH), endogenous regulators of redox; 4-hydroxy-2-nonenal (HNE)-modified protein, a marker of lipid peroxidation; and nuclear factor (NF)-kappaB, a redox sensitive transcription factor in retinal samples, was measured by immunohistochemistry and Western blot or enzymatic recycling assay. Light-induced retinal damage estimated by electroretinography and quantitative immunohistochemistry for 8-hydroxy-2-deoxyguanosine (8OHdG index), a marker of oxidative stress-induced DNA damage, was compared in NAC- and PBS-treated mice. RESULTS: Upregulation of TRX and HNE-modified protein, decrease of GSH, and nuclear translocation of NF-kappaB were noted after light exposure in PBS-treated mice. These changes were suppressed in NAC-treated mice compared with PBS-treated mice. GRX was not upregulated after light exposure in any mice. The a- and b-wave amplitudes were significantly higher, and the 8OHdG index was significantly lower after light exposure in NAC-treated mice than in PBS-treated mice. CONCLUSIONS: Retinal redox status is altered by intense light and is normalized partially by the effect of NAC on TRX and GSH tissue levels. Manipulation of the tissue redox state by exogenous thiol replenishment may be a useful strategy to prevent retinal photooxidative damage. PMID- 12091443 TI - Rescue of axotomized retinal ganglion cells by BDNF gene electroporation in adult rats. AB - PURPOSE: To determine whether the brain-derived neurotrophic factor (BDNF) gene can be transfected into retinal ganglion cells (RGCs) by electroporation and whether axotomized RGCs can be rescued after transfection by BDNF in adult rats. METHODS: Mouse BDNF cDNA was injected intravitreally followed by in vivo electroporation in adult rats. The expression of BDNF in RGCs was confirmed by Western immunoblot analysis and immunohistochemistry. After introduction of BDNF cDNA, the survival of axotomized RGCs was estimated by the TdT-dUTP terminal nick end labeling (TUNEL) method and measured by counting the number of RGCs that were labeled retrogradely by 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyamine percholorate (diI) applied to the superior colliculus (SC). RESULTS: Eyes with injection of the BDNF gene followed by in vivo electroporation showed a significantly higher level of expression of BDNF in the RGC layer, a higher rescue ratio, and a lower number of TUNEL-positive cells than the control samples. CONCLUSIONS: These findings demonstrate that electroporation is an effective method for the direct delivery of genes into RGCs, and that the BDNF gene transferred into RGCs by in vivo electroporation can protect axotomized RGCs against apoptosis. PMID- 12091444 TI - An in vivo gene therapy approach for experimental proliferative vitreoretinopathy using the truncated platelet-derived growth factor alpha receptor. AB - PURPOSE: Proliferative vitreoretinopathy (PVR) is a serious problem in vitreoretinal surgeries. A report of a previous study has indicated that platelet derived growth factor alpha receptor (alphaPDGFR) plays an important role in a rabbit model of this disease and that a dominant negative alphaPDGFR potently suppresses PVR in an ex vivo setting. Herein, the effect of in vivo gene delivery of a dominant negative alphaPDGFR on PVR was tested in a rabbit model of the disease. METHODS: The dominant negative alphaPDGFR (TalphaR) is a truncated version of the receptor, which does not have the intracellular domain. It was expressed by using a retrovirus. In vitro characterization of TalphaR was performed in primary cultures of rabbit conjunctival fibroblasts (RCFs). Western blot analysis was used to check the expression of TalphaR protein. A type I collagen gel contraction assay was performed to test the efficacy of TalphaR on PDGF-dependent cellular responses in vitro. The in vivo efficacy and specificity of the retrovirus was determined by injecting a green fluorescent protein (GFP) retrovirus into rabbits that had been preinjected with RCFs. The impact of the TalphaR retrovirus on PVR was tested by using the rabbit model in which PVR was induced by the injection of RCFs and platelet-rich plasma (PRP). RESULTS: TalphaR was expressed at more that 50 times the level of endogenous alphaPDGFR in RCFs and severely reduced PDGF-dependent contraction of collagen gels. Intravitreal injection of the GFP retrovirus resulted in expression of GFP primarily in the injected RCFs. Whereas injection of RCFs induced complete retinal detachment in 100% of the animals, co-injection of the TalphaR retrovirus substantially reduced the severity and incidence of retinal detachments. CONCLUSIONS: Gene therapy with a retrovirus used to express a dominant negative alphaPDGFR attenuates PVR in a rabbit model of the disease. This strategy may be a new approach to preventing PVR in humans. PMID- 12091445 TI - The ability of rapid retinal reattachment to stop or reverse the cellular and molecular events initiated by detachment. AB - PURPOSE: To determine the effects of reattachment on the molecular and cellular events initiated by a retinal detachment lasting 1 hour or 1 day. METHODS: Experimental retinal detachments were created in the right eyes of nine cats. Reattachments were performed 1 hour (n = 3) or 1 day (n = 3) after the detachment, and the animals were killed 3 days after detachment. Three-day detached (n = 3) and normal (n = 3) retinas were used for comparisons. Agarose embedded sections were double labeled with a panel of antibodies. Some sections were also probed with the TUNEL technique to detect apoptotic cells. Wax-embedded sections were labeled with the MIB-1 antibody to the Ki67 protein to detect proliferating cells. RESULTS: The 1-hour and 1-day detachments followed by reattachment showed a very similar and consistent reduction in photoreceptor deconstruction and the Muller cell gliotic response when compared with 3-day retinal detachments without reattachment. Light microscopy and immunolabeling with opsin antibodies showed a significant reduction in both rod and cone outer segment (OS) degeneration, even though OS length was shorter than normal. The reattachments also showed a reduction in opsin redistribution, retraction of rod terminals, TUNEL-labeled photoreceptors, loss of cytochrome oxidase staining in photoreceptors, neurite outgrowth from second-order neurons, the number of proliferating cells, and the increase in intermediate filaments and loss of soluble proteins from Muller cells. The apparent re-ensheathing of the OS by the apical processes of the retinal pigment epithelium had begun but was not completely normal. CONCLUSIONS: These data indicate that, even though the length of the OS is less than normal, retinal reattachment within 1 day of detachment can either greatly retard or reverse many of the molecular and cellular changes initiated by detachment. PMID- 12091446 TI - Molecular steps involved in light-induced oxidative damage to retinal rods. AB - PURPOSE: To define the molecular mechanism underlying light-induced oxidative damage to retinal photoreceptors. METHODS: Oxidative stress was induced in isolated rod photoreceptors by bright 470- to 490-nm light and monitored by measuring the conversion of dihydrorhodamine 123 to rhodamine, with fluorescence microscopy. The effect of the wavelength on oxidant generation was investigated by applying prebleaching stimuli of either 485- or 520-nm light before the bright 470- to 490-nm light. The role of internal messengers in photooxidative stress and membrane damage by bright 470- to 490-nm light was investigated by patch clamp recording. RESULTS: Constant illumination with bright 470- to 490-nm light caused a rapid increase in generation of oxidants, which peaked after approximately 60 seconds, and a decrease in membrane resistance, eventually producing irreversible membrane damage. The time course and extent of oxidant generation were not affected by the absence of intracellular guanosine triphosphate (GTP) or adenosine triphosphate (ATP), suggesting that oxidative stress and membrane damage induced by 470- to 490-nm light do not require coupling to a GTP-binding protein. Prebleaching exposure to 520-nm light suppressed oxidative stress and membrane damage by subsequent application of bright 470- to 490-nm light, and the extent of suppression increased with prebleaching duration. CONCLUSIONS: Oxidative stress and damage induced in rods in response to 470- to 490-nm light require rhodopsin activation, but not visual transduction steps downstream of active rhodopsin. Prebleaching with 485- or 520 nm light has a different effect on the level of a transient rhodopsin intermediate required for lipid peroxidation by 470- to 490-nm light. PMID- 12091447 TI - Regression of ocular neovascularization in response to increased expression of pigment epithelium-derived factor. AB - PURPOSE: Several pharmacologic treatments have been shown to reduce ocular neovascularization when administered before the onset of angiogenic stimuli, but none have been shown to cause regression of already established ocular neovascularization. In this study, the authors tested the effect of adenoviral vectored pigment epithelium-derived factor (PEDF) gene transfer on established neovascularization in transgenic mice with expression of vascular endothelial growth factor (VEGF) in photoreceptors (rho/VEGF mice) and in a model of choroidal neovascularization. METHODS: Two weeks after the onset of VEGF transgene expression in rho/VEGF mice or 2 weeks after laser-induced rupture of Bruch's membrane in wild-type mice, subgroups of mice were killed, and the baseline amount of neovascularization was measured by image analysis. The remainder of the mice received an intravitreous or subretinal injection of adenoviral vector containing a PEDF expression construct (AdPEDF.11) or control vector (AdNull.11). RESULTS: Seven days after injection in rho/VEGF mice or 10 days after injection in the choroidal neovascularization model, the amount of neovascularization in AdPEDF.11-injected eyes was significantly less than the baseline level, indicating that regression of neovascularization had occurred. There was TUNEL staining within choroidal neovascular lesions in eyes injected with AdPEDF.11. Eyes given a subretinal injection of AdNull.11 had TUNEL-positive cells in the retina, but none within areas of choroidal neovascularization. CONCLUSIONS: These data indicate that increased expression of PEDF causes regression of ocular neovascularization by promoting apoptosis of cells within neovascular lesions and possibly represents a new treatment paradigm for patients with established ocular neovascularization. PMID- 12091448 TI - Spectral profiling of autofluorescence associated with lipofuscin, Bruch's Membrane, and sub-RPE deposits in normal and AMD eyes. AB - PURPOSE: To compare the autofluorescence spectra of retinal pigment epithelium (RPE)-associated lipofuscin, Bruch's membrane, and sub-RPE deposits (drusen and basal laminar-linear deposits) in eyes of donors with age-related macular degeneration (AMD) against eyes of age-matched control donors. METHODS: Cryosections were cut from the maculae of unfixed human donor eyes with AMD or from age-matched control eyes. Tissues were excited at wavelengths of 364, 488, 568, and 633 nm. Emission spectra were collected with a confocal microscope equipped with a spectrophotometric detector at 10-nm wavelength intervals between 400 and 800 nm. RESULTS: RPE lipofuscin had strong autofluorescent emissions that were excited at all wavelengths. Bruch's membrane exhibited strong autofluorescence with an emission peak of 485 +/- 5 nm when excited with 364-nm light. At 488-, 568-, and 633-nm excitations, Bruch's membrane and sub-RPE deposits in normal eyes exhibited minimal autofluorescence. In AMD eyes, however, both the 364- and 488-nm excitation wavelengths stimulated substantial blue-green emissions from sub-RPE deposits and Bruch's membrane, with average pixel intensities substantially exceeding that elicited in the yellow-orange range by RPE lipofuscin. CONCLUSIONS: These data suggest that an increase in blue-green autofluorescence of Bruch's membrane relative to the yellow-orange autofluorescence of RPE-associated lipofuscin is associated with AMD. Knowledge of these spectra will be useful in evaluating animal models of macular degenerative disease and in diagnosis of AMD, and will provide a novel signature for further analysis of the molecular entities emitting these fluorescent signatures. PMID- 12091449 TI - The role of c-fos in cell death and regeneration of retinal ganglion cells. AB - PURPOSE: To investigate the effect of c-fos on apoptotic cell death and regeneration of damaged retinal ganglion cells (RGCs) in tissue culture of retinal explants. METHODS: Retinas from transgenic mice carrying the exogenous c fos gene under the control of the interferon (IFN)-alpha/beta inducible Mx promoter (Mx-c-fos), c-fos-deficient mice, and littermate control mice were dissected and cultured in a three-dimensional collagen gel culture system, followed by an analysis of TdT-dUTP terminal nick-end labeling (TUNEL) staining and measurement of neurites that emerged from explants. RESULTS: Compared with littermate control mice, Mx-c-fos transgenic animals showed a higher ratio of TUNEL positivity in the RGC layer from early in the culture period that correlated with the small number of regenerating neurites. In contrast, the c-fos null mutated mice showed a still-lower ratio of TUNEL-positive cells. Nevertheless, the number of regenerating neurites was significantly lower in the initial phase, although the drastic increase in density of neurite regeneration was observed in the late period of culture. CONCLUSIONS: These findings suggest that c-fos is involved in both apoptotic cell death and regeneration of damaged RGCs. Elucidation of the precise c-fos-mediated cascade involved in RGC apoptosis and regeneration is significant in realizing neuronal survival and regeneration. PMID- 12091450 TI - Lysophosphatidic acid receptor signaling in mammalian retinal pigment epithelial cells. AB - PURPOSE: Lysophosphatidic acid (LPA) is a phospholipid growth factor that stimulates proliferation, chemotaxis, cation currents, and K(+) currents in retinal pigment epithelial (RPE) cells. LPA receptor transduction was analyzed in human and rat RPE cells. METHODS: Cells were cultured with standard methods, and signaling pathways were analyzed with a variety of approaches, including whole cell recording, calcium imaging, and second-messenger assays. RESULTS: LPA activated nonselective cation currents in rat RPE were blocked by the protein tyrosine kinase (PTK) inhibitor genistein, by the MAP kinase kinase (MEK) inhibitor PD98059, and by loading cells with antibodies to G(alpha(i)/o/t/z). LPA activated the MAP kinase and extracellular signal-related kinase (ERK)-1, and produced a dose-dependent inhibition of cAMP production. LPA stimulated a dose dependent increase in [Ca(2+)](i) that persisted in Ca(2+)-free medium and was reduced by pretreatment with thapsigargin, suggesting it involves release from intracellular stores. The [Ca(2+)](i) increase was not blocked by ryanodine or the phospholipase C inhibitor U73122. LPA did not stimulate inositol phosphate production. Similar to the cation current, LPA-evoked [Ca(2+)](i) increases were blocked by PD98059 and by loading cells with antibodies to G(alpha(i)/o/t/z). RT PCR experiments showed the presence of RNA for three LPA receptor subtypes (Edg2, -4, and -7); RNase protection assays showed the strongest expression for Edg2 receptor RNA. CONCLUSIONS: LPA receptors in RPE cells activate pertussis toxin (PTx)-sensitive G proteins that inhibit cAMP accumulation; stimulate MAP kinase which activates a cation current and probably contributes to mitogenesis; and stimulate release of Ca(2+) from intracellular stores that appears independent of IP(3) and ryanodine receptor activation. PMID- 12091451 TI - Quantitative assessment of the integrity of the blood-retinal barrier in mice. AB - PURPOSE: The purpose of this study was to develop and characterize a quantitative assay of blood-retinal barrier (BRB) function in mice and to determine the effect of several purported vasopermeability factors on the BRB. METHODS: Adult C57BL/6J mice were treated with three regimens of increasingly extensive retinal cryopexy and subsequently were given an intraperitoneal injection of 1 microCi/g body weight of [(3)H]mannitol. At several time points, the amount of radioactivity per milligram tissue was compared in retina, lung, and kidney. Time points that maximize signal-to-background differential in the retina were identified, and the ratio of counts per minute (CPM) per milligram retina to CPM per milligram lung (retina-to-lung leakage ratio, RLLR) or kidney (retina-to-renal leakage ratio, RRLR) were calculated. This technique was then used to compare the amount of BRB breakdown that occurs after intravitreous injection of vascular endothelial growth factor (VEGF), insulin-like growth factor (IGF)-1, prostaglandin (PG) E(1), PGE(2), interleukin (IL)-1beta, or tumor necrosis factor (TNF)-alpha. RESULTS: Twenty-four hours after retinal cryopexy, there was a higher level of radioactivity in treated than in control retinas, and the signal-to-background difference was optimal when measurements were obtained 1 hour after injection of [(3)H]mannitol. In untreated mice, the RLLR was 0.30 +/- 0.02 and the RRLR was 0.22 +/- 0.01. Twenty-four hours after one 5-second application of retinal cryopexy, the RLLR was 0.73 +/- 0.20 and the RRLR was 0.71 +/- 0.23. With increasing amounts of cryopexy, there was an increase in the RLLR and RRLR, so that after two 10-second applications, the RLLR was 1.66 +/- 0.31 and the RRLR was 1.47 +/- 0.20. Intravitreous injection of VEGF, IGF-1, PGE(1), PGE(2), IL 1beta, or TNF-alpha each caused significant increases in the RLLR and RRLR, but there were some differences in potency and time course. VEGF caused prominent BRB breakdown at 6 hours that returned to near normal by 24 hours. IL-1beta also caused relatively rapid breakdown of the BRB, but its effect was more prolonged than that caused by VEGF. There was delayed, but substantial breakdown of the BRB after injection of TNF-alpha. IGF-1, PGE(2), and PGE(1) caused less severe, relatively delayed, and more prolonged BRB breakdown. CONCLUSIONS: Measurement of the RLLR or RRLR after intraperitoneal injection of [(3)H]mannitol in mice provides a quantitative assessment of BRB function that is normalized and can therefore be compared from assay to assay. Comparison of the extent and duration of BRB breakdown after intravitreous injection of vasoactive substances shows that agents can be grouped by resultant extent and time course of leakage. Additional studies are needed to determine whether this grouping has its basis in shared mechanisms of BRB disruption. PMID- 12091452 TI - Visual pigment coexpression in all cones of two rodents, the Siberian hamster, and the pouched mouse. AB - PURPOSE: To decide whether the identical topography of short- and middle wavelength cone photoreceptors in two species of rodents reflects the presence of both opsins in all cone cells. METHODS: Double-label immunocytochemistry using antibodies directed against short-wavelength (S)-and middle- to long-wavelength (M/L)-sensitive opsin were used to determine the presence of visual pigments in cones of two species of rodents, the Siberian hamster (Phodopus sungorus) and the pouched mouse (Saccostomus campestris) from South Africa. Topographical distribution was determined from retinal whole-mounts, and the colocalization of visual pigments was examined using confocal laser scanning microscopy. Opsin colocalization was also confirmed in consecutive semithin tangential sections. RESULTS: The immunocytochemical results demonstrate that in both the Siberian hamster and the pouched mouse all retinal cones contain two visual pigments. No dorsoventral gradient in the differential expression of the two opsins is observed. CONCLUSIONS: The retina of the Siberian hamster and the pouched mouse is the first example to show a uniform coexpression of M and S cone opsins in all cones, without any topographical gradient in opsin expression. This finding makes these two species good models for the study of molecular control mechanisms in opsin coexpression in rodents, and renders them suitable as sources of dual cones for future investigations on the role and neural connections of this cone type. PMID- 12091453 TI - Vitreous treatment of cultured human RPE cells results in differential expression of 10 new genes. AB - PURPOSE: To analyze the differential gene expression in cultured human retinal pigment epithelial (RPE) cells after treatment with vitreous. METHODS: Cultured human RPE cells were incubated for 48 hours with 25% human vitreous from donor eyes. Total RNA from treated and untreated cells was extracted. The gene expression was analyzed by differential expression analysis (DEmRNA-PCR). The differentially expressed genes were identified by gene bank searches. Differential expression was verified by a quantitative real-time RT-PCR fluorescent nucleic acid staining system. The in vivo mRNA expression of these genes in RPE cells was shown by gene-specific RT-PCR. RESULTS: Vitreous treatment of human RPE cells resulted in the reduced expression of NFIB2, KE03 (NY-REN 25ag), PIG-B, DKFZp564BC462, LKHA, G3BP, PAM, UEV-1, and MAP1B calibrated to the expression of GAPDH when compared with their expression in untreated cells. The reduced expression after vitreous treatment was quantified by gene-specific quantitative real-time RT-PCR and varied from 0.69 to 0.17 compared with untreated cells. The mRNA expression of UDP-GalNac mRNA remained constant. The mRNA expression of eight of these genes was demonstrated in this study for the first time in human RPE cells. CONCLUSIONS: Vitreous treatment of cultured RPE cells induces the differential expression of a variety of genes with functions in transcription, mediation of signal transduction and inflammation, glycosylation, ubiquitination and protein-protein interaction. Further examination of these genes may locate additional targets for treatment of diseases caused by contact of RPE cells with vitreous, typical in proliferative vitreoretinopathy. PMID- 12091454 TI - Evidence for a brief period of enhanced oxygen susceptibility in the rat model of oxygen-induced retinopathy. AB - PURPOSE: Findings in a previous study have shown that the retina of newborn rats exposed to hyperoxia during the first days of life sustain permanent functional (as determined with the rod ERG) and structural (as determined with histology) damage that appears to be determined by the level of retinal maturity reached at the time of oxygen exposure-the retinas of rat pups being more susceptible to hyperoxic shock during the second week of life than during the first week. Given that the cone ERG has been shown to mature later than the rod ERG, the purpose of the present study was to examine whether cone responses also demonstrates a similar maturational susceptibility to postnatal hyperoxia. Also examined was whether the oscillatory potentials (OPs) were affected by postnatal hyperoxia. METHODS: Newborn rats were exposed to hyperoxia during selected postnatal day intervals either initiated at birth (early-onset exposure) or at a later postnatal age (late-onset exposure). Photopic and scotopic (mixed cone-rod) electroretinograms were recorded at 30 days. RESULTS: Data analysis reveals that photopic and scotopic responses (b-wave and OPs) demonstrated a similar maturational susceptibility to postnatal hyperoxia, in which exposure regimens initiated during the second week of life were most detrimental to retinal function. The results also revealed a temporal window of enhanced oxygen susceptibility at approximately postnatal day 10. The duration of this window was longer when estimated with the scotopic responses, but the extent of the functional damage was more pronounced when estimated with the photopic signals. Finally, compared with the b-wave, the OPs, especially the short-latency OPs, were proportionally more affected. CONCLUSIONS: The results suggest that cone function is significantly more susceptible to postnatal hyperoxia than rod function, and the OPs appear to be the most susceptible ERG components, thus suggesting a differential susceptibility to oxygen toxicity of the different retinal components. However, despite a clear demonstration of its existence, the exact nature of the temporal window of enhanced oxygen susceptibility as well as a possible equivalence in other animal models of oxygen induced retinopathy, including the human form (retinopathy of prematurity), remains to be determined. PMID- 12091455 TI - Correlating retinal function with melatonin secretion in subjects with an early or late circadian phase. AB - PURPOSE: Evaluate the diurnal variation of retinal function, as measured with the electroretinogram (ERG), in subjects with an early (morning type: M-type) or a late (evening type: E-type) circadian phase. METHODS: Subjects (n = 24) were recruited according to their scores on a Morningness-Eveningness Questionnaire assessing preferences in, e.g., bedtime, waketime, and timing of performance. ERG testing was performed twice on each subject, at 22:30 and at 08:00. Luminance response functions were obtained in scotopic (blue flashes) and in photopic conditions (white, blue, green, and red flashes). Salivary melatonin samples were taken every half-hour from 20:30 to 00:00 and from 06:30 to 09:30. RESULTS: In scotopic conditions, both groups had lower ERG amplitudes and retinal sensitivity at 08:00. In photopic conditions, the two groups showed an opposite pattern of diurnal variations. The E-types demonstrated a significant reduction in ERG amplitudes at 08:00, whereas the M-types showed an increase in amplitude at the same time. In addition, negative correlations were found between both the cone ERG and mixed rod-cone ERG and the concentration of salivary melatonin, indicating that the ERG amplitude is lowest when melatonin concentration is highest. CONCLUSIONS: The reduction in scotopic ERG responses at 08:00 seen in both groups might be due to the peak of rod disc shedding that takes place, in some mammals, at around light onset. The strong correlation between the cone ERG and salivary melatonin could be attributable to a direct effect of retinal melatonin on the physiology of cones or of the circadian phase of the subjects. PMID- 12091456 TI - Post-photoreceptoral activity dominates primate photopic 32-Hz ERG for sine-, square-, and pulsed stimuli. AB - PURPOSE: To evaluate the relative contributions of photoreceptors and postphotoreceptoral activity to the primate 32-Hz flicker electroretinogram (ERG) elicited by sine-wave, square-wave, and pulse stimuli. METHODS: Flicker 32-Hz ERGs were evoked from four adult rhesus (Macaca mulatta) monkeys using sine-wave, square-wave, and 4-ms pulse trains and xenon photostrobe flicker stimuli. All stimuli had time-averaged luminance of 2.11 log cd/m(2) and were presented on a 1.63-log cd/m(2) white background. Intravitreal injections of DL-2-amino-4 phosphonobutyric acid (APB) and cis-2,3-piperidinedicarboxylic acid (PDA) were given to block activity of ON- and OFF-bipolar cells, respectively. RESULTS: Flicker harmonic analysis showed that the fundamental frequency component provided nearly 75% of the sine-wave and square-wave ERGs versus 63% for 4-ms pulse stimuli and only 49% for strobe flicker. Strobe-flicker responses contained the greatest contribution from higher harmonic components. Removing the ON component with APB increased the fundamental component's amplitudes by more than 30% with sine-wave and square-wave ERGs but had a lesser effect on responses to 4 ms pulses and strobe flicker. When cone responses were isolated by synaptic blockade with APB+PDA, the fundamental component's amplitude was reduced to less than 20% of control for all four stimuli. Postsynaptic ON and OFF components were characterized by amplitude and phase vectors, and sine-wave and square-wave stimuli gave a large phase difference (138 degrees ) between ON and OFF components, which resulted in greater response self-cancellation than with the 4 ms pulse train (121 degrees phase difference) or for strobe flicker (118 degrees ). CONCLUSIONS: The major decrease in flicker responses after photoreceptor synaptic blockade implicates a major contribution from postphotoreceptoral activity to the fundamental flicker component, regardless of the stimulus type. Sine-wave and square-wave stimuli produced larger phase differences between ON- and OFF-pathway components, thereby causing more complete self-cancellation of postphotoreceptoral contributions and revealing slightly greater relative contribution directly from cone photoreceptors with these stimuli than with pulsed stimuli. The direct cone contribution was always small, however, and the clinical point is that 32-Hz flicker ERG amplitudes do not provide an unambiguous assessment of direct cone photoreceptor contribution with any of these stimuli. PMID- 12091459 TI - The binding of prion proteins to serum components is affected by detergent extraction conditions. AB - As many GPI anchored proteins, PrP(C) and its abnormal conformer PrP(Sc), are inserted into membrane microdomains known as rafts. Upon raft disruption, PrP(C) becomes soluble, while PrP(Sc) aggregates into insoluble structures. It was recently published that, as opposed to PrP(C), PrP(Sc), as well as its protease resistant core PrP27-30, can bind specifically to plasminogen and other serum components. These findings were suggested to have important physiological implications in transmissible spongiform encephalopathies (TSE) diagnosis and pathogenesis. In this work, we show that the binding of PrP(Sc) or PrP 27-30 to serum proteins occurs only at specific detergent combinations, in which disease associated PrPs are present in aggregated structures. At detergent conditions in which rafts are intact, it is actually PrP(C.) that binds to blood proteins, albeit not directly, but through neighboring rafts components. Our results therefore indicate that the binding of PrP(Sc) to blood components has no physiological relevance. PMID- 12091460 TI - One melanocortin 4 and two melanocortin 5 receptors from zebrafish show remarkable conservation in structure and pharmacology. AB - We report the cloning, genome mapping, functional expression, pharmacology and anatomical distribution of three melanocortin (MC) receptors from zebrafish (z). Phylogenetic analysis showed with high bootstrap support that these genes represent one MC4 receptor and two MC5 receptors. Chromosomal mapping showed conserved synteny between regions containing zMC4 and human (h) MC4 receptors, whereas the two zMC5 receptor genes map on chromosome segments in which the zebrafish has several genes with two orthologues of a single mammalian gene. It is likely that the two copies of zMC5 receptors arose through a separate duplication in the teleost lineage. The zMC4, zMC5a, and zMC5b receptors share 70 71% overall amino acid identity with the respective human orthologues and over 90% in three TM regions believed to be most important for ligand binding. All three zebrafish receptors also show pharmacological properties remarkably similar to their human orthologues, with similar affinities and the same potency order, when expressed and characterized in radioligand binding assay for the natural MSH) peptides alpha-, beta-, and gamma-MSH. Stimulation of transfected mammalian cells with alpha-MSH caused a dose-dependent increase in intracellular cAMP levels for all three zebrafish receptors. All three genes were expressed in the brain, eye, ovaries and gastrointestinal tract, whereas the zMC5b receptor was also found in the heart, as determined by RT-PCR. Our studies, which represent the first characterization of MC receptors in a nonamniote species, indicate that the MC receptor subtypes arose very early in vertebrate evolution. Important pharmacological and functional properties, as well as gene structure and syntenic relationships have been highly conserved over a period of more than 400 million years implying that these receptors participate in vital physiological functions. PMID- 12091457 TI - Pupil location under mesopic, photopic, and pharmacologically dilated conditions. AB - PURPOSE: To determine whether there are systematic changes in pupil location with changes in the state of pupil size and with other ocular variables. METHODS: High resolution images of the pupil of the eyes of 70 subjects were taken using an infrared-sensitive camera. Images were obtained under mesopic, photopic, and pharmacologically dilated conditions. From the images, the center and diameter of the corneal limbus and the pupil were computed. In addition, the location of the first Purkinje image was calculated. RESULTS: The pupil center shifted consistently temporally as the pupil dilated. The total motion was relatively small, with a mean distance of 0.133 mm motion between the mesopic and photopic conditions, with the pupil diameter changing from 6.3 to 4.1 mm. Ninety percent of the subjects had a motion of less than 0.3 mm. One patient showed a motion of almost 0.6 mm. The change in location of the pupil center was not significantly related to refractive error, age, or the change of pupil diameter. CONCLUSIONS: Changes in the location of the pupil center with changes in the dilation of the pupil are typically slight, but can be significant in a few subjects, especially in pharmacologically dilated pupils. PMID- 12091461 TI - Poly(ADP-ribose) polymerase inhibition prevents both apoptotic-like delayed neuronal death and necrosis after H(2)O(2) injury. AB - Toxic reactive oxygen species (ROS) such as hydrogen peroxide, nitric oxide, superoxide, and the hydroxyl radical are generated in a variety of neuropathological conditions and cause significant DNA damage. We determined the effects of 3-aminobenzamide (AB), an inhibitor of the DNA repair enzyme poly(ADP ribose) polymerase (PARP), on cell death in differentiated PC12 cells, a model of sympathetic neurons, after H(2) O(2) injury. Exposure to 0.5 mm H(2) O(2) resulted in a significant decrease in intracellular NAD(H), NADP(H), and ATP levels. This injury resulted in the death of 90% of the cells with significant necrosis early (2 h) after injury and increased apoptosis (12-24 h after injury), as measured by PS exposure and the presence of cytoplasmic oligonucleosomal fragments. Treatment with 2.5 mm AB restored pyridine nucleotide and ATP levels and ameliorated cell death (65% versus 90%) by decreasing the extent of both necrosis and apoptosis. Interestingly, we observed that H(2) O(2) -induced injury caused a delayed cell death exhibiting features of apoptosis but in which caspase 3 like activity was absent. Moreover, pretreatment with AB restored caspase-3 like activity. Our results suggest that apoptosis and necrosis are both triggered by PARP overactivation, and that maintenance of cellular energy levels after injury by inhibiting PARP shifts cell death from necrosis to apoptosis. PMID- 12091462 TI - 1-Methyl-4-phenylpyridinium ion, a toxin that can cause parkinsonism, alters branched structures of DNA. AB - During replication, human mitochondrial DNA (mtDNA) takes on a triple-stranded structure known as a D-loop, which is implicated in replication and transcription. 1-Methyl-4-phenylpyridinium ion (MPP+), a toxin inducing parkinsonism, inhibits mtDNA replication, possibly by resolving the D-loops. For initiation of mtDNA replication, mitochondria are thought to have another triple stranded structure, an R-loop. The R-loop, which is resolved by a bacterial junction-specific helicase, RecG, is also resolved by MPP+. Because mitochondrial D-loops are likewise resolved by RecG, the D- and R-loops may share a similar branched structure. MPP+ resolves cruciform DNA in supercoiled DNA. MPP+ converts a stacked conformation to an extended conformation in a synthetic Holliday junction. This conversion is reversed by 1 mM Mg(2+), as is the resolution of the D-loops or cruciform DNA. These observations suggest that the junction structure of mitochondrial D- and R-loops is affected by MPP+. PMID- 12091463 TI - Effect of ethanol on protein kinase Czeta and p70S6 kinase activation by carbachol: a possible mechanism for ethanol-induced inhibition of glial cell proliferation. AB - The signal transduction pathways that mediate the mitogenic response of muscarinic acetylcholine receptors in astroglial cells have not been fully elucidated. In this study we investigated the activation of p70S6 kinase (p70S6K) by carbachol in 1321 N1 astroctyoma cells. Carbachol induced a dose- and time dependent activation of p70S6K, as evidenced by increased phosphorylation at Thr 389, Thr-421 and Ser-424, by increased p70S6K activity, and by a shift in its molecular weight. Activation of p70S6K was mediated by M3 muscarinic acetylcholine receptors (mAChRs) and was inhibited by two phosphatidylinositol-3 kinase (PI3-K) inhibitors, by a pseudosubstrate to protein kinase C (PKC) zeta, and by the p70S6K inhibitor rapamycin. Carbachol-induced DNA synthesis was strongly inhibited by rapamycin, suggesting that p70S6K activation plays an important role in carbachol-induced cell proliferation. Ethanol (25-100 mm) has been shown to inhibit carbachol-induced proliferation of astroglial cells. In the same range of concentrations, ethanol also inhibits carbachol-induced activation of PKCzeta and of p70S6K. On the other hand, inhibition of PI3-kinase was only observed at higher ethanol concentrations. These results indicate that activation of the PKCzeta--> p70S6K pathway by M3 mAChRs may play a role in the increased DNA synthesis and may represent a target for ethanol-induced inhibition of astroglial cell proliferation. PMID- 12091464 TI - Time-dependence of the contribution of pyruvate carboxylase versus pyruvate dehydrogenase to rat brain glutamine labelling from [1-(13) C]glucose metabolism. AB - [1-(13) C]glucose metabolism in the rat brain was investigated after intravenous infusion of the labelled substrate. Incorporation of the label into metabolites was analysed by NMR spectroscopy as a function of the infusion time: 10, 20, 30 or 60 min. Specific enrichments in purified mono- and dicarboxylic amino acids were determined from (1) H-observed/(13) C-edited and (13) C-NMR spectroscopy. The relative contribution of pyruvate carboxylase versus pyruvate dehydrogenase (PC/PDH) to amino acid labelling was evaluated from the enrichment difference between either C2 and C3 for Glu and Gln, or C4 and C3 for GABA, respectively. No contribution of pyruvate carboxylase to aspartate, glutamate or GABA labelling was evidenced. The pyruvate carboxylase contribution to glutamine labelling varied with time. PC/PDH decreased from around 80% after 10 min to less than 30% between 20 and 60 min. This was interpreted as reflecting different labelling kinetics of the two glutamine precursor glutamate pools: the astrocytic glutamate and the neuronal glutamate taken up by astrocytes through the glutamate-glutamine cycle. The results are discussed in the light of the possible occurrence of neuronal pyruvate carboxylation. The methods previously used to determine PC/PDH in brain were re-evaluated as regards their capacity to discriminate between astrocytic (via pyruvate carboxylase) and neuronal (via malic enzyme) pyruvate carboxylation. PMID- 12091465 TI - Glycine transporter isoforms show differential subcellular localization in PC12 cells. AB - The subcellular localization of glycine transporters one (GLYT1) and two (GLYT2) stably expressed in PC12 cells has been studied. To facilitate visualization, enhanced green fluorescent protein (GFP) was fused to the amino terminus of both glycine transporters. Functional analysis of the GFP-GLYT1 and GFP-GLYT2 stable cell lines demonstrated that they exhibited high affinity for glycine and the characteristic properties of both glycine transporter subtypes. The GFP-coupled transporters were differently distributed throughout the cell. GFP-GLYT1 was mainly localized on the plasma membrane, whereas most of GFP-GLYT2 was present on large dense-core vesicles and endosomes. Both transporters were absent from the synaptic vesicle population in PC12 cells. PMID- 12091466 TI - Inhibition of brain mitochondrial respiration by dopamine: involvement of H(2)O(2) and hydroxyl radicals but not glutathione-protein-mixed disulfides. AB - Examination of the downstream mediators responsible for inhibition of mitochondrial respiration by dopamine (DA) was investigated. Consistent with findings reported by others, exposure of rat brain mitochondria to 0.5 mm DA for 15 min at 30 degrees C inhibited pyruvate/glutamate/malate-supported state-3 respiration by 20%. Inhibition was prevented in the presence of pargyline and clorgyline demonstrating that mitochondrial inhibition arose from products formed following MAO metabolism and could include hydrogen peroxide (H(2) O(2) ), hydroxyl radical, oxidized glutathione (GSSG) or glutathione-protein mixed disulfides (PrSSG). As with DA, direct incubation of intact mitochondria with H(2) O(2) (100 microm) significantly inhibited state-3 respiration. In contrast, incubation with GSSG (1 mm) had no effect on O(2) consumption. Exposure of mitochondria to 1 mm GSSG resulted in a 3.3-fold increase in PrSSG formation compared with 1.4- and 1.5-fold increases in the presence of 100 microm H(2) O(2) or 0.5 mm DA, respectively, suggesting a dissociation between PrSSG formation and effects on respiration. The lack of inhibition of respiration by GSSG could not be accounted for by inadequate delivery of GSSG into mitochondria as increases in PrSSG levels in both membrane-bound (2-fold) and intramatrix (3.5-fold) protein compartments were observed. Furthermore, GSSG was without effect on electron transport chain activities in freeze-thawed brain mitochondria or in pig heart electron transport particles (ETP). In contrast, H(2) O(2) showed differential effects on inhibition of respiration supported by different substrates with a sensitivity of succinate > pyruvate/malate > glutamate/malate. NADH oxidase and succinate oxidase activities in freeze-thawed mitochondria were inhibited with IC(50) approximately 2-3-fold higher than in intact mitochondria. ETPs, however, were relatively insensitive to H(2) O(2). Co-administration of desferrioxamine with H(2) O(2) had no effect on complex I-associated inhibition in intact mitochondria, but attenuated inhibition of rotenone-sensitive NADH oxidase activity by 70% in freeze-thawed mitochondria. The results show that DA associated inhibition of respiration is dependent on MAO and that H(2) O(2) and its downstream hydroxyl radical rather than increased GSSG and subsequent PrSSG formation mediate the effects. PMID- 12091467 TI - BDNF-mediated signal transduction is modulated by GSK3beta and mood stabilizing agents. AB - Brain-derived neurotrophic factor (BDNF) is a major neurotrophin in the brain and abnormal regulation of BDNF may contribute to the pathophysiology of mood disorders. In the present study, we examined if alterations in the activity of glycogen synthase kinase-3-beta (GSK3beta) or treatment with mood stabilizers modulated BDNF-mediated signal transduction pathways in differentiated human neuroblastoma SH-SY5Y cells. BDNF increased the phosphorylation of the forkhead transcription factor FKHRL1 through activation of the phosphatidylinositol 3 kinase (PI3K)/Akt signaling pathway, and the phosphorylation of the cyclic AMP response element binding protein (CREB) through activation of extracellular signal-regulated kinase1/2 (ERK1/2). BDNF also increased serine(9) phosphorylation of GSK3beta, which inhibits GSK3beta activity. Overexpression of GSK3beta did not affect BDNF-induced phosphorylation of Akt, ERK1/2, or FKHRL1, but abolished CREB phosphorylation induced by BDNF. This inhibition of BDNF induced CREB phosphorylation in GSK3beta-overexpressing SH-SY5Y cells was blocked by treatment with lithium. In contrast to lithium, sodium valproate and lamotrigine did not affect BDNF-mediated signaling, whereas carbamazepine induced a rapid and prolonged phosphorylation of ERK1/2 and CREB in the absence or the presence of BDNF. Therefore, increased GSK3beta selectively attenuates BDNF induced CREB phosphorylation, and lithium and carbamazepine can facilitate activation of CREB. PMID- 12091469 TI - Stimulation of 5-hydroxytryptamine (5-HT(2C) ) receptors in the ventrotegmental area inhibits stress-induced but not basal dopamine release in the rat prefrontal cortex. AB - The present study investigated whether 5-HT(2C) receptors in the ventrotegmental area and prefrontal cortex regulate basal and stimulus-evoked dopamine release in the prefrontal cortex. Using the in vivo microdialysis technique in conscious rats, we studied the effect of a selective 5-HT(2C) receptor agonist, Ro60-0175, on basal and immobilization stress-induced dopamine release in the prefrontal cortex. Ro60-0175 intraperitoneally (2.5 mg/kg) and into the ventrotegmental area (10 microg/0.5 microL) completely antagonized the effect of stress on extracellular dopamine without altering basal levels. Infusion of 10 microm Ro60 0175 through the cortical probe had no significant effect on basal and stress induced dopamine release. SB242084 (10 mg/kg), a selective antagonist of 5-HT(2C) receptors, significantly increased basal extracellular dopamine and completely prevented the effect of intraperitoneal and intraventrotegmental Ro60-0175 on the stress-induced rise of extracellular dopamine, but had no effect itself in stressed rats. The results show that Ro60-0175 suppresses cortical dopamine release induced by immobilization stress through the stimulation of 5-HT(2C) receptors in the ventrotegmental area. While confirming that endogenous 5-HT acting on 5-HT(2C) receptors tonically inhibit basal dopamine release in the prefrontal cortex, the present findings suggest that the stimulation of 5-HT(2C) receptors with an exogenous agonist preferentially inhibit stimulated release. PMID- 12091468 TI - Comparison of huntingtin proteolytic fragments in human lymphoblast cell lines and human brain. AB - Proteolytic fragments of huntingtin (htt) in human lymphoblast cell lines from HD and control cases were compared to those in human HD striatal and cortical brain regions, by western blots with epitope-specific antibodies. HD lymphoblast cell lines were heterozygous and homozygous for the expanded CAG triplet repeat mutations, which represented adult onset and juvenile HD. Lymphoblasts contained NH(2)- and COOH-terminal htt fragments of 20-100 kDa, with many similar htt fragments in HD compared to control lymphoblast cell lines. Detection of htt fragments in a homozygous HD lymphoblast cell line demonstrated proteolysis of mutant htt. It was of interest that adult HD lymphoblasts showed a 63-64 kDa htt fragment detected by the NH(2)-domain antibody, which was not found in controls. In addition, control and HD heterozygous cells showed a common 60-61 kDa band (detected by the NH(2)-domain antibody), which was absent in homozygous HD lymphoblast cells. These results suggest that the 63-64 kDa and 60-61 kDa NH(2) domain htt fragments may be associated with mutant and normal htt, respectively. In juvenile HD lymphoblasts, the presence of a 66-kDa, instead of the 63-64 kDa N domain htt fragment, may be consistent with the larger polyglutamine expansion of mutant htt in the juvenile case of HD. Lymphoblasts and striatal or cortical regions from HD brains showed similarities and differences in NH(2)- and COOH terminal htt fragments. HD striatum showed elevated levels of 50 and 45 kDa NH(2) terminal htt fragments [detected with anti(1-17) serum] compared to controls. Cortex from HD and control brains showed similar NH(2)-terminal htt fragments of 50, 43, 40, and 20 kDa; lymphoblasts also showed NH(2)-terminal htt fragments of 50, 43, 40, and 20 kDa. In addition, a 48-kDa COOH-terminal htt band was elevated in HD striatum, which was also detected in lymphoblasts. Overall, results demonstrate that mutant and normal htt undergo extensive proteolysis in lymphoblast cell lines, with similarities and differences compared to htt fragments observed in HD striatal and cortical brain regions. These data for in vivo proteolysis of htt are consistent with the observed neurotoxicity of recombinant NH(2)-terminal mutant htt fragments expressed in transgenic mice and in transfected cell lines that may be related to the pathogenesis of HD. PMID- 12091470 TI - Neuronal transfer of the human Cu/Zn superoxide dismutase gene increases the resistance of dopaminergic neurons to 6-hydroxydopamine. AB - Several mechanisms are thought to be involved in the progressive decline in neurons of the substantia nigra pars compacta (SNpc) that leads to Parkinson's disease (PD). Neurotoxin 6-hydroxydopamine (6-OHDA), which induces parkinsonian symptoms in experimental animals, is thought to be formed endogenously in patients with PD through dopamine (DA) oxidation and may cause dopaminergic cell death via a free radical mechanism. We therefore investigated protection against 6-OHDA by inhibiting oxidative stress using a gene transfer strategy. We overexpressed the antioxidative Cu/Zn-superoxide dismutase (SOD1) enzyme in primary culture dopaminergic cells by infection with an adenovirus carrying the human SOD1 gene (Ad-hSOD1). Survival of the dopaminergic cells exposed to 6-OHDA was 50% higher among the SOD1-producing cells than the cells infected with control adenoviruses. In contrast, no significant increased survival of (6-OHDA) treated dopaminergic cells was observed when they were infected with an adenovirus expressing the H(2) O(2) -scavenging glutathione peroxidase (GPx) enzyme. These results underline the major contribution of superoxide in the dopaminergic cell death process induced by 6-OHDA in primary cultures. Overall, this study demonstrates that the survival of the dopaminergic neurons can be highly increased by the adenoviral gene transfer of SOD1. An antioxidant gene transfer strategy using viral vectors expressing SOD1 is therefore potentially beneficial for protecting dopaminergic neurons in PD. PMID- 12091471 TI - Association of protein kinase C with GABA(A) receptors containing alpha1 and alpha4 subunits in the cerebral cortex: selective effects of chronic ethanol consumption. AB - Previous studies have suggested that protein kinase C (PKC) isoforms differentially influence the sensitivity of gamma-aminobutyric acid(A) (GABA(A) ) receptor responses in brain. Both PKCgamma and PKCepsilon knock-out mice exhibit altered ethanol potentiation of GABA(A) receptor mediated Cl(-) flux. Furthermore, chronic ethanol consumption alters GABA(A) receptor function and receptor subunit peptide expression by mechanisms that are not yet understood. The present study explored the possibility that PKC isoforms are directly associated with GABA(A) receptors, and this association is influenced by chronic ethanol exposure. GABA(A) receptors containing alpha1 or alpha4 subunits were immunoprecipitated from solubilized protein derived from the membrane fraction of rat cerebral cortex using selective antibodies. Immunoprecipitated receptors were screened by western blot analysis for the presence of PKCdelta, gamma and epsilon isoforms. We found pronounced labeling of PKCgamma but not PKCdelta or PKCepsilon in the alpha1 and alpha4 subunit immunoprecipitates. Immunoprecipitation with PKCgamma, but not with IgG antibody also yielded GABA(A) receptor alpha1 and alpha4 subunits in the immunoprecipitate. The association of PKCgamma with alpha1 containing receptors was decreased 44 +/- 11% after chronic ethanol consumption. In contrast, PKCgamma associated with alpha4-containing receptors was increased 32 +/- 7% after chronic ethanol consumption. These results suggest that PKCgamma may be involved in GABA(A) receptor adaptations following chronic ethanol consumption. PMID- 12091472 TI - Disruptive effects of glucocorticoids on glutathione peroxidase biochemistry in hippocampal cultures. AB - Glucocorticoids (GCs), the adrenal steroids secreted during stress, compromise the ability of hippocampal neurons to survive various necrotic insults. We have previously observed that GCs enhance the hippocampal neurotoxicity of reactive oxygen species and, as a potential contributor to this, decrease the activity of the antioxidant enzyme, glutathione peroxidase (GSPx). In this report, we have studied the possible mechanisms underlying this GC effect upon GSPx in primary hippocampal cultures and have observed several results. (i) Corticosterone (the GC of rats) decreased glutathione levels; this was predominately a result of a decrease in levels of reduced glutathione (GSH), the form of glutathione which facilitates GSPx activity. (ii) Corticosterone also decreased levels of NADPH; this may help explain the effect on GSH as NADPH is required for regeneration of GSH from oxidized glutathione. (iii) However, the corticosterone effect on total glutathione levels could not just be caused by the NADPH effect, as there were also reduced levels of oxidized glutathione. (iv) Corticosterone caused a small but significant decrease in GSPx activity over a range of glucose concentrations; this occurred under circumstances of an excess of glutathione as a substrate, suggesting a direct effect of corticosterone on GSPx activity. (v) This corticosterone effect was likely to have functional implications, in that enhancement of GSPx activity (to the same magnitude as activity was inhibited by corticosterone) by GSPx overexpression protected against an excitotoxin. Thus, GCs have various effects, both energetic and non-energetic in nature, upon steps in GSPx biochemistry that, collectively, may impair hippocampal antioxidant capacity. PMID- 12091473 TI - Alterations in metabolism and gene expression in brain regions during cuprizone induced demyelination and remyelination. AB - Exposure of mice to the copper chelator, cuprizone, results in CNS demyelination. There is remyelination after removal of the metabolic insult. We present brain regional studies identifying corpus callosum as particularly severely affected; 65% of cerebroside is lost after 6 weeks of exposure. We examined recovery of cerebroside and ability to synthesize cerebroside and cholesterol following removal of the toxicant. The temporal pattern for concentration of myelin basic protein resembled that of cerebroside. We applied Affymetrix GeneChip technology to corpus callosum to identify temporal changes in levels of mRNAs during demyelination and remyelination. Genes coding for myelin structural components were greatly down-regulated during demyelination and up-regulated during remyelination. Genes related to microglia/macrophages appeared in a time-course (peaking at 6 weeks) correlating with phagocytosis of myelin and repair of lesions. mRNAs coding for many cytokines had peak expression at 4 weeks, compatible with intercellular signaling roles. Of interest were other genes with temporal patterns correlating with one of the three above patterns, but of function not obviously related to demyelination/remyelination. The ability to correlate gene expression with known pathophysiological events should help in elucidating further function of such genes as related to demyelination/remyelination. PMID- 12091474 TI - Lithium inhibits aluminum-induced apoptosis in rabbit hippocampus, by preventing cytochrome c translocation, Bcl-2 decrease, Bax elevation and caspase-3 activation. AB - A variety of studies on neuronal death models suggest that lithium has neuroprotective properties. In the present investigation, we have examined the effect of chronic lithium treatment on hippocampus, as monitored by changes at the subcellular level of apoptosis-regulatory proteins which have been induced by the neurotoxin, aluminum maltolate. Intracisternal administration of aluminum into rabbit brain induces cytochrome c release, decreases levels of the anti apoptotic proteins Bcl-2 and Bcl-X(L), increases levels of the pro-apoptotic Bax, activates caspase-3, and causes DNA fragmentation as measured by the TUNEL assay. Pretreatment for 14 days with 7 mm of lithium carbonate in drinking water prevents aluminum-induced translocation of cytochrome c, and up-regulates Bcl-2 and Bcl-X(L,) down-regulates Bax, abolishes caspase-3 activity and reduces DNA damage. The regulatory effect of lithium on the apoptosis-controlling proteins occurs in both the mitochondria and endoplasmic reticulum. We propose that the neuroprotective effect of lithium involves the modulation of apoptosis-regulatory proteins present in the subcellular organelles of rabbit brain. PMID- 12091475 TI - Effects of desipramine treatment on norepinephrine transporter gene expression in the cultured SK-N-BE(2)M17 cells and rat brain tissue. AB - The antidepressant desipramine (DMI) is a selective inhibitor of norepinephrine (NE) transport that down-regulates the norepinephrine transporter (NET) protein in a concentration- and time-dependent manner in vitro. In this study, possible regulatory effects of DMI on NET mRNA and protein levels were investigated with the NET-expressing SK-N-BE(2)M17 cell line and rat brain tissue. Northern blot analysis showed that incubation of the cultured cells with DMI (5-500 nm) for 3 days reduced levels of NET mRNA in both its 5.8-kb (by up to 58%) and 3.6-kb forms (to 68%), whereas incubation for 14 days increased both levels (to 40% and 100%) in a concentration-dependent manner. In contrast, NET protein levels decreased after 3-14 days of exposure of the cells to DMI, as determined by western blotting. The in vitro findings were supported by in vivo treatment of rats with DMI. Thus, in situ hybridization demonstrated initially decreased, and later increased, NET mRNA levels in locus coeruleus (LC) tissue of rats treated with DMI; whereas NET protein levels in the LC were reduced after 14 days, but unchanged after three daily DMI treatments. Thus, DMI had similar effects on NET expression in vitro and in vivo, with opposite changes in NET mRNA and protein levels, suggesting that the regulatory mechanisms involved are complex and non congruent. PMID- 12091476 TI - Blockade of cannabinoid CB(1) receptor function protects against in vivo disseminating brain damage following NMDA-induced excitotoxicity. AB - The ability of cannabinoid CB(1) receptors to influence glutamatergic excitatory neurotransmission has fueled interest in how these receptors and their endogenous ligands may interact in conditions of excitotoxic insults. The present study characterized the impact of stimulated and inhibited CB(1) receptor function on NMDA-induced excitotoxicity. Neonatal (6-day-old) rat pups received a systemic injection of a mixed CB(1) /CB(2) receptor agonist (WIN55,212-2) or their respective antagonists (SR141716A for CB(1) and SR144528 for CB(2) ) prior to an unilateral intrastriatal microinjection of NMDA. The NMDA-induced excitotoxic damage in the ipsilateral forebrain was not influenced by agonist-stimulated CB(1) receptor function. In contrast, blockade of CB(1), but not CB(2), receptor activity evoked a robust neuroprotective response by reducing the infarct area and the number of cortical degenerating neurons. These results suggest a critical involvement of CB(1) receptor tonus on neuronal survival following NMDA receptor induced excitotoxicity in vivo. PMID- 12091477 TI - Repression of myelin proteolipid protein gene expression is mediated through both general and cell type-specific negative regulatory elements in nonexpressing cells. AB - The myelin proteolipid protein gene (Plp ) is expressed primarily in oligodendrocytes. Yet how the gene remains repressed in nonexpressing cells has not been defined, and potentially could cause adverse effects in an organism if the mechanism for repression was impaired. Previous studies suggest that the first intron contains element(s), which suppress expression in nonexpressing cells, although the identity of these elements within the 8 kb intron was not characterized. Here we report the localization of multiple negative regulatory elements that repress Plp gene expression in nonexpressing cells (+/+ Li). Two of these elements (regions) correspond to those used by Plp expressing cells (N20.1), whilst another acts in a cell type-specific manner (i.e. operational in +/+ Li liver cells, but not N20.1 cells). By gel-shift and DNase I footprinting analyses, the factor(s) that bind to the cell type-specific negative regulatory region appear to be far more abundant in +/+ Li cells than in N20.1 cells. Thus, Plp gene repression is mediated through the combinatorial action of both "general" and cell type-specific negative regulatory elements. Additionally, repression in +/+ Li cells cannot be overcome via an antisilencer/enhancer element, which previously has been shown to function in N20.1 cells. PMID- 12091478 TI - Convergent evidence from microdialysis and presynaptic immunolabeling for the regulation of gamma-aminobutyric acid release in the globus pallidus following acute clozapine or haloperidol administration in rats. AB - Antipsychotic drugs (APDs) have been primarily characterized for their effects on dopaminergic terminal regions in the brain, especially within the corpus striatum. Efferent GABA pathways are the primary outflow of striatal processing via their projections to the substantia nigra and the globus pallidus (GP). In the current study, we analyzed changes in pallidal GABA function following acute APD administration by means of in vivo microdialysis, followed by immunolabeling of presynaptic GABA terminal density in the contralateral hemisphere of the same animals. Acute administration of the atypical APD, clozapine (10 or 30 mg/kg, s.c.), produced a dose-dependent decrease in extracellular GABA. A corresponding dose-dependent increase in the density of presynaptic terminal GABA immunolabeling in the GP was found. In contrast, the typical APD, haloperidol (1 or 3 mg/kg, s.c.), had no significant effects on either measure, although a non significant increase in extracellular GABA and decrease in the density of GABA terminal immunolabeling was noted. Paw retraction tests conducted during the time of microdialysis showed that haloperidol produced a typical pattern of highly pronounced motor impairment, while clozapine showed an atypical profile of minimal catalepsy. These complementary results obtained from in vivo neurochemistry and presynaptic neurotransmitter labeling suggest that systemic clozapine suppresses neuronal GABA release within the GP. This decrease in released pallidal GABA may play a role in the low motor side-effect liability of atypical APDs. PMID- 12091479 TI - Intranuclear localization of apoptosis-inducing factor (AIF) and large scale DNA fragmentation after traumatic brain injury in rats and in neuronal cultures exposed to peroxynitrite. AB - Programmed cell death occurs after ischemic, excitotoxic, and traumatic brain injury (TBI). Recently, a caspase-independent pathway involving intranuclear translocation of mitochondrial apoptosis-inducing factor (AIF) has been reported in vitro; but whether this occurs after acute brain injury was unknown. To address this question adult rats were sacrificed at various times after TBI. Western blot analysis on subcellular protein fractions demonstrated intranuclear localization of AIF in ipsilateral cortex and hippocampus at 2-72 h. Immunocytochemical analysis showed AIF labeling in neuronal nuclei with DNA fragmentation in the ipsilateral cortex and hippocampus. Immunoelectronmicroscopy verified intranuclear localization of AIF in hippocampal neurons after TBI, primarily in regions of euchromatin. Large-scale DNA fragmentation ( approximately 50 kbp), a signature event in AIF-mediated cell death, was detected in ipsilateral cortex and hippocampi by 6 h. Neuron-enriched cultures exposed to peroxynitrite also demonstrated intranuclear AIF and large-scale DNA fragmentation concurrent with impaired mitochondrial respiration and cell death, events that are inhibited by treatment with a peroxynitrite decomposition catalyst. Intranuclear localization of AIF and large-scale DNA fragmentation occurs after TBI and in neurons under conditions of oxidative/nitrosative stress, providing the first evidence of this alternative mechanism by which programmed cell death may proceed in neurons after brain injury. PMID- 12091482 TI - The lipase gene family. AB - Development of the lipase gene family spans the change in science that witnessed the birth of contemporary techniques of molecular biology. Amino acid sequencing of enzymes gave way to cDNA cloning and gene organization, augmented by in vitro expression systems and crystallization. This review traces the origins and highlights the functional significance of the lipase gene family, overlaid on the background of this technical revolution. The gene family initially consisted of three mammalian lipases [pancreatic lipase (PL), lipoprotein lipase, and hepatic lipase] based on amino acid sequence similarity and gene organization. Family size increased when several proteins were subsequently added based on amino acid homology, including PL-related proteins 1 and 2, phosphatidylserine phospholipase A1, and endothelial lipase. The physiological function of each of the members is discussed as well as the region responsible for lipase properties such as enzymatic activity, substrate binding, heparin binding, and cofactor interaction. Crystallization of several lipase gene family members established that the family belongs to a superfamily of enzymes, which includes esterases and thioesterases. This superfamily is related by tertiary structure, rather than amino acid sequence, and represents one of the most populous families found in nature. PMID- 12091481 TI - The mutation of two amino acid residues in the N-terminus of tyrosine hydroxylase (TH) dramatically enhances the catalytic activity in neuroendocrine AtT-20 cells. AB - The sequence Arg37-Arg38 of tyrosine hydroxylase (TH) is known to play a significant role in the feedback inhibition by the end product DA. To clarify how deeply the sequence Arg37-Arg38 and the phosphorylated Ser40 of human TH type 1 (hTH1) are involved in the regulation of this feedback inhibition in mammalian cells, we generated the following mutants: (i) RR-GG, Arg37-Arg38 replaced by Gly37-Gly38; (ii) RR-EE, Arg37-Arg38 replaced by Glu37-Glu38; (iii) S40D, Ser40 replaced by Asp40; and (iv) S40A, Ser40 replaced by Ala40. In a cell-free system, the level of the DA inhibition of the RR-EE mutant enzyme was to the same or smaller degree than that of the phosphorylation-mimicking S40D. Next, AtT-20 neuroendocrine cells were transfected with wild-type and mutated TH genes because these cells were earlier shown to be capable of fully converting L-3,4 dihydroxyphenylalanine into DA, whereby the catalytic activity of TH would be expected to be inhibited by the end product DA accumulating in the cells. The level of DA accumulation in AtT-20 cells expressing the TH gene was in the order: RR-EE > S40D > S40A = RR-GG > wild-type, which was in accordance with the observations for the cell-free system. These results suggest that the sequence Arg37-Arg38 of hTH1 is a more potent determinant of the efficient production of DA in mammalian cells than is the phosphorylated Ser40-hTH1. PMID- 12091480 TI - The role of muscarinic acetylcholine receptor-mediated activation of extracellular signal-regulated kinase 1/2 in pilocarpine-induced seizures. AB - Pilocarpine-induced seizures are mediated by the M(1) subtype of muscarinic acetylcholine receptor (mAChR), but little is known about the signaling mechanisms linking the receptor to seizures. The extracellular signal-regulated kinase (ERK) signaling cascade is activated by M(1) mAChR and is elevated during status epilepticus. Yet, the role of ERK activation prior to seizure has not been evaluated. Here, we examine the role of pilocarpine-induced ERK activation in the induction of seizures in mice by pharmacological and behavioral approaches. We show that pilocarpine induces ERK activation prior to the induction of seizures by both western blot and immunocytochemistry with an antibody to phosphorylated ERK. In addition, we show that the ERK pathway inhibitor SL327 effectively blocks the pilocarpine-induced ERK activation. However, SL327 pretreatment has no effect on the initiation of seizures. In fact, animals treated with SL327 had higher seizure-related mortality than vehicle-treated animals, suggesting activated ERK may serve a protective role during seizures. In addition, ERK inhibition had no effect on the development of the long-term sequelae of status epilepticus (SE), including mossy fiber sprouting, neuronal death and spontaneous recurrent seizures. PMID- 12091483 TI - Interactions of very long-chain saturated fatty acids with serum albumin. AB - The remarkable binding properties of serum albumin have been investigated extensively, but little is known about an important class of fatty acids, the very long-chain saturated fatty acids (VLCFA; >18 carbons). Although VLCFA are metabolized efficiently in normal individuals, they are markers for and possibly causative agents of several peroxisomal disorders. We studied the binding of [(13)C]carboxyl-enriched arachidic (C20:0), behenic (C22:0), lignoceric (C24:0), and hexacosanoic (C26:0) acids to bovine serum albumin (BSA) by (13)C-NMR spectroscopy. For each VLCFA, the NMR spectra showed multiple signals at chemical shifts previously identified for long-chain fatty acids (12-18 carbons), suggesting stabilization of binding by similar, if not identical, interactions of the fatty acid carboxyl anion with basic amino acid residues. The maximal binding (mol of VLCFA/mol of BSA) and the number of observed binding sites decreased with increasing chain length, from 4-5 for C20:0, 3-4 for C22:0, and 2 for C24:0; we validated our previous conclusion that BSA has only one site for C26:0 (Ho, J. K., H. Moser, Y. Kishimoto, and J. A. Hamilton. 1995. J. Clin. Invest. 96: 1455 1463). Analysis of chemical shifts suggested that the highest affinity sites for VLCFA are low affinity sites for long-chain fatty acids. In competition experiments with (13)C-labeled C22:0 (3 mol/mol of BSA) and unlabeled oleic acid, C22:0 bound to BSA in the presence of up to 4 mol of oleic acid/mol of BSA, but 1 mol was shifted into a different site. Our studies suggest that albumin has adequate binding capacity for the low plasma levels of VLCFA with 20 to 26 carbons, but the protein may not be able to bind longer chain VLCFA. PMID- 12091484 TI - Two novel missense mutations in the CETP gene in Japanese hyperalphalipoproteinemic subjects: high-throughput assay by Invader assay. AB - Cholesteryl ester transfer protein (CETP) deficiency is one of the most important and common causes of hyperalphalipoproteinemia (HALP) in the Japanese. CETP deficiency is thought to be a state of impaired reverse cholesterol transport, which may possibly lead to the development of atherosclerotic cardiovascular disease despite high HDL-cholesterol (HDL-C) levels. Thus, it is important to investigate whether HALP is caused by CETP deficiency. In the present study, we identified two novel missense mutations in the CETP gene among 196 subjects with a marked HALP (HDL-C > or = 2.59 mmol/l = 100 mg/dl). The two missense mutations, L151P (CTC-->CCC in exon 5) and R282C (CGC-->TGC in exon 9), were found in compound heterozygous subjects with D442G mutation, whose plasma CETP levels were significantly lower when compared with those in D442G heterozygous subjects. In COS-7 cells expressing the wild type and mutant CETP, these two mutant CETP showed a marked reduction in the secretion of CETP protein into media (0% and 39% of wild type for L151P and R282C, respectively). These results suggested that two novel missense mutations cause the decreased secretion of CETP protein into circulation leading to HALP. By using the Invader assay for seven mutations, including two novel mutations of the CETP gene, we investigated their frequency among 466 unrelated subjects with HALP (HDL-C > or = 2.07 mmol/l = 80 mg/dl). Two novel mutations were rare, but L151P mutation was found in unrelated subjects with a marked HALP. Furthermore, we demonstrated that CETP deficiency contributes to 61.7% and 31.4% of marked HALP and moderate HALP in the Japanese, respectively. PMID- 12091486 TI - Sources and properties of triglyceride-rich lipoproteins containing apoB-48 and apoB-100 in postprandial blood plasma of patients with primary combined hyperlipidemia. AB - Although editing of apolipoprotein (apo)B in the small intestine, yielding apoB 48, is thought to be nearly complete in adult humans, small amounts of intestinal apoB-100 may also be produced. We have evaluated the fraction of unedited apoB secreted from the intestine postprandially in subjects with primary combined hyperlipidemia, a disorder in which secretion of apoB-100 into the blood is increased. Three hours after these subjects and healthy controls were fed a fat rich meal containing retinol, the distribution of retinyl esters (RE) between plasma triglyceride-rich lipoprotein (TRL) fractions containing apoB-100 and apoB 48 was measured under conditions minimizing transfer of RE between lipoprotein particles. The estimated maximal percentage of unedited intestinal apoB-100 (approximately 3%) was not increased in subjects with primary combined hyperlipidemia, suggesting that reduced editing of intestinal mRNA does not contribute to the pathogenesis of this disorder. Postprandially, the triglyceride content of TRL containing apoB-48 more than doubled, leading to a 20% increase in mean diameter, yet the surface concentration of phospholipids and soluble apolipoproteins (apoE and total apoC) was unchanged. Furthermore, the surface concentrations of these components did not differ among TRL containing apoB-48 and two smaller fractions of apoB-100 TRL with distinct immunoreactivities. These findings suggest that available surface area is a major determinant of the particle content of each of these surface components of TRL species of differing size and origin. PMID- 12091485 TI - Association of GM4 ganglioside with the membrane surrounding lipid droplets in shark liver. AB - By TLC, GM4 was found to be the major ganglioside in the liver of six shark species examined: Odontaspis taurus, Negaprion brevirostris, Sphyrna lewini, Mustelus griseus, Mustelus manazo, and Prionace glauca. A detailed analysis of the glycosphingolipids (GSLs) in the liver of O. taurus (sand tiger shark) showed that it contained approximately 110 nmol of lipid-bound sialic acid per gram of wet tissue, of which 80% was GM4. By extracting the liver of O. taurus with chloroform/methanol, followed by chromatographic separation of GSLs using DEAE Sephadex A-25 and Iatrobeads columns, we have isolated GM4 in pure form with a yield of approximately 5 mg per 100 g of wet tissue. The structures of both the sugar chain and the ceramide moiety of this GM4 were analyzed by chemical analysis, mass spectrometry, and NMR spectroscopy. Similar to GM4 isolated from other sources, 92% of fatty acids in the ceramide of this GM4 were 2 hydroxylated. However, unlike the long-chain bases found in other GSLs, the total long-chain bases in this GM4 were found to contain 43% octadecasphingenine and 50% nonadecasphingenine. Immunohistochemical analysis using a monoclonal antibody against GM4 revealed that the hepatocytes of both M. griseus (spotless smooth hound) and M. manazo (smooth hound) were filled with lipid droplets and GM4 was primarily associated with the membrane structure surrounding lipid droplets. PMID- 12091487 TI - Inhibition of phosphatidylcholine synthesis via the phosphatidylethanolamine methylation pathway impairs incorporation of bulk lipids into VLDL in cultured rat hepatocytes. AB - Inhibition of phosphatidylcholine (PC) synthesis via the phosphatidylethanolamine (PE) methylation pathway was shown to decrease the secretion of VLDL from primary rat hepatocytes (Nishimaki-Mogami et al. 1996. BIOCHIM: Biophys. Acta. 1304: 21 31). To understand further the role of PE methylation, we determined the effect of bezafibrate, an inhibitor of PE methylation, on VLDL assembly within the microsomal lumen. Bezafibrate was shown to decrease VLDL (triacylglycerol) secretion only when cellular PE methylation was active in the presence of methionine. Pulse-chase experiments showed that bezafibrate treatment did not impair the movement of [(35)S]apolipoprotein (apo)B-48 from microsomal membranes into the lumen. However, bezafibrate treatment resulted in reduced VLDL [(35)S]apoB-48 and increased [(35)S]apoB-48-containing particles in the HDL density range (HDL-[(35)S]apoB-48) within the lumen. Inhibition of PE methylation by bezafibrate or 3-deazaadenosine after the completion of HDL-[(35)S]apoB-48 assembly effectively decreased VLDL-[(35)S]apoB-48 secretion with a concomitant increase in HDL-[(35)S]apoB-48 secretion. These findings suggest that inhibition of PC synthesis via the PE methylation pathway impairs the stage of bulk triacylglycerol incorporation during the assembly of VLDL. PMID- 12091488 TI - Incorporation of cholesterol in sphingomyelin- phosphatidylcholine vesicles has profound effects on detergent-induced phase transitions. AB - Vesicle <--> micelle transitions are important phenomena during bile formation and intestinal lipid processing. The hepatocyte canalicular membrane outer leaflet contains appreciable amounts of phosphatidylcholine (PC) and sphingomyelin (SM), and both phospholipids are found in the human diet. Dietary SM enrichment inhibits intestinal cholesterol absorption. We therefore studied detergent-induced vesicle --> micelle transitions in SM-PC vesicles. Phase transitions were evaluated by spectrophotometry and cryotransmission electron microscopy (cryo-TEM) after addition of taurocholate (3-7 mM) to SM-PC vesicles (4 mM phospholipid, SM/PC 40%/60%, without or with 1.6 mM cholesterol). After addition of excess (5-7 mM) taurocholate, SM-PC vesicles were more sensitive to micellization than PC vesicles. As shown by sequential cryo-TEM, addition of equimolar (4 mM) taurocholate to SM-PC vesicles induced formation of open vesicles, then (at the absorbance peak) fusion of bilayer fragments into large open structures (around 200 nm diameter) coexisting with some multilamellar or fused vesicles and thread-like micelles and, finally, transformation into an uniform picture with long thread-like micelles. Incorporation of cholesterol in the SM/PC bilayer changed initial vesicular shape from spherical into ellipsoid and profoundly increased detergent resistance. Disk-like micelles and multilamellar vesicles, and then extremely large vesicular structures, were observed by sequential cryo-TEM under these circumstances, with persistently increased absorbance values by spectrophotometry. These findings may be relevant for bile formation and intestinal lipid processing. Inhibition of intestinal cholesterol absorption by dietary SM enrichment may relate to high resistance against bile salt-induced micellization of intestinal lipids in presence of the sphingolipid. PMID- 12091489 TI - LXR/RXR activation enhances basolateral efflux of cholesterol in CaCo-2 cells. AB - Regulation of gene expression of ATP-binding cassette transporter (ABC)A1 and ABCG1 by liver X receptor/retinoid X receptor (LXR/RXR) ligands was investigated in the human intestinal cell line CaCo-2. Neither the RXR ligand, 9-cis retinoic acid, nor the natural LXR ligand 22-hydroxycholesterol alone altered ABCA1 mRNA levels. When added together, ABCA1 and ABCG1 mRNA levels were increased 3- and 7 fold, respectively. T0901317, a synthetic non-sterol LXR agonist, increased ABCA1 and ABCG1 gene expression 11- and 6-fold, respectively. ABCA1 mass was increased by LXR/RXR activation. T0901317 or 9-cis retinoic acid and 22-hydroxycholesterol increased cholesterol efflux from basolateral but not apical membranes. Cholesterol efflux was increased by the LXR/RXR ligands to apolipoprotein (apo)A I or HDL but not to taurocholate/phosphatidylcholine micelles. Actinomycin D prevented the increase in ABCA1 and ABCG1 mRNA levels and the increase in cholesterol efflux induced by the ligands. Glyburide, an inhibitor of ABCA1 activity, attenuated the increase in basolateral cholesterol efflux induced by T0901317. LXR/RXR activation decreased the esterification and secretion of cholesterol esters derived from plasma membranes. Thus, in CaCo-2 cells, LXR/RXR activation increases gene expression of ABCA1 and ABCG1 and the basolateral efflux of cholesterol, suggesting that ABCA1 plays an important role in intestinal HDL production and cholesterol absorption. PMID- 12091490 TI - Native LDL potentiate TNF alpha and IL-8 production by human mononuclear cells. AB - Native LDL (nLDL) increases expression of adhesion molecules on endothelial cells through induction of Ca(2+) mobilization. Ca(2+) mobilization is also involved in the induction of proinflammatory cytokines, important mediators involved in atherogenesis. The aim of the study was to evaluate the capacity of nLDL to affect spontaneous and lipopolysaccharide (LPS)-stimulated cytokine production. Preincubation of human peripheral blood mononuclear cells (PBMC) with nLDL for 24 h did not influence spontaneous production of tumor necrosis factor alpha (TNF alpha) or interleukin-8 (IL-8), but significantly potentiated LPS-induced production of these cytokines. nLDL preincubation of PBMC did not increase the expression of the LPS receptors Toll-like receptor-4, CD14, or CD11c/CD18. Potentiation of cytokine production by nLDL was mediated through induction of Ca(2+) mobilization, because: a) nLDL induced a sustained pattern of repetitive Ca(2+) transients in human PBMC; b) the Ca(2+) chelator fura 2-acetoxymethyl ester, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, an intracellular Ca(2+) chelator, inhibited the potentiating effect of nLDL on LPS-induced cytokine synthesis; c) induction of Ca(2+) mobilization by thapsigargin potentiated LPS-induced cytokine production. nLDL are able to potentiate LPS induced production of cytokines by human PBMC, and this effect is probably mediated through induction of Ca(2+) mobilization. This may represent an important pathogenetic mechanism in atherogenesis induced by hyperlipoproteinemia. PMID- 12091491 TI - Serum plant sterols and biliary cholesterol secretion in humans: studies with ursodeoxycholic acid. AB - Ratios of cholestanol, campesterol, and sitosterol to cholesterol in serum are known to reflect cholesterol absorption efficiency. Here, a possible link between these ratios and biliary secretion rates of cholesterol was investigated. Biliary lipid secretion rates and serum sterols were determined in 13 patients with gallstones. Seven were treated with ursodeoxycholic acid (UDCA) (1,000 mg/d). Serum cholesterol and non-cholesterol sterols were also measured in a cross over study in 20 healthy volunteers, who received either placebo or UDCA (750 mg/d). Biliary cholesterol secretion was significantly lower, whereas the non cholesterol sterols and their ratio to cholesterol were higher in patients with gallstones treated with UDCA. A highly significant negative linear correlation between the ratios of non-cholesterol sterols to cholesterol and biliary cholesterol secretion was observed. In volunteers, administration of UDCA for 4 weeks was followed by a significant increase in non-cholesterol sterols and their ratios. Even 4 weeks after discontinuing UDCA administration, campesterol and sitosterol were still significantly higher than pretreatment levels, which was also true for the campesterol-cholesterol ratio after 8 weeks. The results suggest that the ratios of cholestanol, campesterol, and sitosterol to cholesterol can be used as indicators of changes in biliary cholesterol secretion rates. PMID- 12091492 TI - Profile of cholesterol-related sterols in aged amyloid precursor protein transgenic mouse brain. AB - Cholesterol is implicated to play a role in Alzheimer disease pathology. Therefore, the concentrations of cholesterol, its precursors, and its degradation products in brain homogenates of aging wild-type and beta-amyloid precursor protein transgenic mice carrying the Swedish mutation (APP23) were analyzed. Among the sterols measured, lanosterol is the first common intermediate of two different pathways, which use either desmosterol or lathosterol as the predominant precursors for de novo synthesis of brain cholesterol. In young mice, cholesterol is mainly synthesized via the desmosterol pathway, while in aged mice, lathosterol is the major precursor. 24S-hydroxycholesterol (cerebrosterol), which plays a key role in the removal of cholesterol from the brain, modestly increased during aging. No differences in the levels of cholesterol, its precursors, or its metabolites were found between wild-type and APP23 transgenic mice. Moreover, the levels of the exogenous plant sterols campesterol and sitosterol were significantly elevated in the brains of APP23 animals at age 12 and 18 months. This time point coincides with abundant plaque formation. PMID- 12091493 TI - Carotenoid uptake and secretion by CaCo-2 cells: beta-carotene isomer selectivity and carotenoid interactions. AB - In presence of oleate and taurocholate, differentiated CaCo-2 cell monolayers on membranes were able to assemble and secrete chylomicrons. Under these conditions, both cellular uptake and secretion into chylomicrons of beta-carotene (beta-C) were curvilinear, time-dependent (2-16 h), saturable, and concentration-dependent (apparent K(m) of 7-10 microM) processes. Under linear concentration conditions at 16 h incubation, the extent of absorption of all-trans beta-C was 11% (80% in chylomicrons), while those of 9-cis- and 13-cis-beta-C were significantly lower (2-3%). The preferential uptake of the all-trans isomer was also shown in hepatic stellate HSC-T6 cells and in a cell-free system from rat liver (microsomes), but not in endothelial EAHY cells or U937 monocyte-macrophages. Moreover, extents of absorption of alpha-carotene (alpha-C), lutein (LUT), and lycopene (LYC) in CaCo 2 cells were 10%, 7%, and 2.5%, respectively. Marked carotenoid interactions were observed between LYC/beta-C and beta-C/alpha-C. The present results indicate that beta-C conformation plays a major role in its intestinal absorption and that cis isomer discrimination is at the levels of cellular uptake and incorporation into chylomicrons. Moreover, the kinetics of cellular uptake and secretion of beta-C, the inhibition of the intestinal absorption of one carotenoid by another, and the cellular specificity of isomer discrimination all suggest that carotenoid uptake by intestinal cells is a facilitated process. PMID- 12091494 TI - Defects in degradation of blood group A and B glycosphingolipids in Schindler and Fabry diseases. AB - Skin fibroblast cultures from patients with inherited lysosomal enzymopathies, alpha-N-acetylgalactosaminidase (alpha-NAGA) and alpha-galactosidase A deficiencies (Schindler and Fabry disease, respectively), and from normal controls were used to study in situ degradation of blood group A and B glycosphingolipids. Glycosphingolipids A-6-2 (GalNAc (alpha 1-->3)[Fuc alpha 1- >2]Gal(beta1-->4)GlcNAc(beta 1-->3)Gal(beta 1--> 4)Glc (beta 1-->1')Cer, IV(2) alpha-fucosyl-IV(3)-alpha-N-acetylgalactosaminylneolactotetraosylceramide), B-6-2 (Gal(alpha 1-->3)[Fuc alpha 1--> 2] Gal (beta 1-->4)GlcNAc(beta 1-->3)Gal(beta 1- >4)Glc(beta 1-->1')Cer, IV(2)- alpha-fucosyl-IV(3)-alpha galactosylneolactotetraosylceramide), and globoside (GalNAc(beta 1-->3)Gal(alpha 1-->4)Gal(beta 1-->4)Glc(beta 1-->1') Cer, globotetraosylceramide) were tritium labeled in their ceramide moiety and used as natural substrates. The degradation rate of glycolipid A-6-2 was very low in fibroblasts of all the alpha-NAGA deficient patients (less than 7% of controls), despite very heterogeneous clinical pictures, ruling out different residual enzyme activities as an explanation for the clinical heterogeneity. Strongly elevated urinary excretion of blood group A glycolipids was detected in one patient with blood group A, secretor status (five times higher than upper limit of controls), in support of the notion that blood group A-active glycolipids may contribute as storage compounds in blood group A patients. When glycolipid B-6-2 was fed to alpha galactosidase A-deficient cells, the degradation rate was surprisingly high (50% of controls), while that of globotriaosylceramide was reduced to less than 15% of control average, presumably reflecting differences in the lysosomal enzymology of polar glycolipids versus less-polar ones. Relatively high-degree degradation of substrates with alpha-D-Galactosyl moieties hints at a possible contribution of other enzymes. PMID- 12091495 TI - Cholesterol gallstone formation in overweight mice establishes that obesity per se is not linked directly to cholelithiasis risk. AB - The relationship between obesity and cholesterol cholelithiasis is not well understood at physiologic or genetic levels. To clarify whether obesity per se leads to increased prevalence of cholelithiasis, we examined cholesterol gallstone susceptibility in three polygenic (KK/H1J, NON/LtJ, NOD/LtJ) and five monogenic [carboxypeptidase E (Cpe (fat)), agouti yellow (A(y)), tubby (tub), leptin (Lep(ob)), leptin receptor (Lepr (db))] murine models of obesity during ingestion of a lithogenic diet containing dairy fat, cholesterol, and cholic acid. At 8 weeks on the diet, one strain of polygenic obese mice was resistant whereas the others revealed low or intermediate prevalence rates of cholelithiasis. Monogenic obese mice showed distinct patterns with either high or low gallstone prevalence rates depending upon the mutation. Dysfunction of the leptin axis, as evidenced by the Lep(ob) and the Lepr (db) mutations, markedly reduced gallstone formation in a genetically susceptible background strain, indicating that in mice with this genetic background, physiologic leptin homeostasis is a requisite for cholesterol cholelithogenesis. In contrast, the Cpe (fat) mutation enhanced the prevalence of cholelithiasis markedly when compared with the background strain. Since CPE converts many prohormones to hormones, a deficiency of biologically active cholecystokinin is a likely contributor to enhanced susceptibility to cholelithiasis through compromising gallbladder contractility and small intestinal motility. Because some murine models of obesity increased, whereas others decreased cholesterol gallstone susceptibility, we establish that cholesterol cholelithiasis in mice is not simply a secondary consequence of obesity per se. Rather, specific genes and distinct pathophysiological pathways are responsible for the shared susceptibility to both of these common diseases. PMID- 12091496 TI - Comprehensive and definitive structural identities of Pneumocystis carinii sterols. AB - Pneumocystis causes a type of pneumonia in immunodeficient mammals, such as AIDS patients. Mammals cannot alkylate the C-24 position of the sterol side chain, nor can they desaturate C-22. Thus, the reactions leading to these sterol modifications are particularly attractive targets for the development of drugs against fungal and protozoan pathogens that make them. In the present study, the definitive structures of 43 sterol molecular species in rat-derived Pneumocystis carinii were elucidated by nuclear magnetic resonance spectroscopy. Ergosterol, Delta(5,7) sterols, trienes, and tetraenes were not among them. Most (32 of the 43) were 24-alkylsterols, products of S-adenosyl-L-methionine:C-24 sterol methyl transferase (SAM:SMT) enzyme activity. Their abundance is consistent with the suggestion that SAM:SMT is highly active in this organism and that the enzyme is an excellent anti-Pneumocystis drug target. In contrast, the comprehensive analysis strongly suggest that P. carinii does not form Delta(22) sterols, thus C 22 desaturation does not appear to be a drug target in this pathogen. The lanosterol derivatives, 24-methylenelanost-8-en-3 beta-ol and (Z)-24 ethylidenelanost-8-en-3 beta-ol (pneumocysterol), previously identified in human derived Pneumocystis jiroveci, were also detected among the sterols of the rat derived P. carinii organisms. PMID- 12091497 TI - Modulation by nutrients and drugs of liver acyl-CoAs analyzed by mass spectrometry. AB - The profile of liver acyl-CoAs induced by dietary fats of variable compositions or by xenobiotic hypolipidemic amphipathic carboxylates was evaluated in vivo using a novel electrospray ionization tandem mass spectrometry methodology of high resolution, sensitivity, and reliability. The composition of liver fatty acyl-CoAs was found to reflect the composition of dietary fat. Treatment with hypolipidemic carboxylates resulted in liver dominant abundance of their respective acyl-CoAs accompanied by an increase in liver fatty acyl-CoAs. Cellular effects exerted by dietary fatty acids and/or xenobiotic carboxylic drugs may be transduced in vivo by their respective acyl-CoAs. PMID- 12091498 TI - HPLC-MS/MS analysis of the products generated from all-trans-retinoic acid using recombinant human CYP26A. AB - Two mammalian hCYP26A expression systems have been used to analyze the metabolic products of CYP26A. Through the use of extensive HPLC, UV spectroscopy, and liquid chromatography/tandem mass spectrometry (LC-MS/MS) methodology, we have conclusively demonstrated that the complex mixture of products comprises 4-OH-all trans-retinoic acid, 4-oxo-all-trans-retinoic acid, and 18-OH-all-trans-retinoic acid, and more polar products, partially identified as dihydroxy and mono-oxo, mono-hydroxy derivatives. These more polar products are presumed to result from multiple hydroxylations on the beta-ionone ring. The inter-relationship of initial and polar metabolites was inferred from both gene-dose and time-course experiments. Both initial and secondary metabolic steps after 4-oxo-all-trans retinoic acid are ketoconazole-sensitive, suggesting that steps in the production of water-soluble metabolites are cytochrome P450-dependent. PMID- 12091499 TI - Retinoid quantification by HPLC/MS(n). AB - Retinoic acid (RA) mediates most of the biological effects of vitamin A that are essential for vertebrate survival. It acts through binding to receptors that belong to the nuclear receptor transcription factor superfamily (Mangelsdorf et al. 1994). It is also a highly potent vertebrate teratogen. To determine the function and effects of endogenous and exogenous RA, it is important to have a highly specific, sensitive, accurate, and precise analytical procedure. Current analyses of RA and other retinoids are labor intensive, of poor sensitivity, have limited specificity, or require compatibility with RA reporter cell lines (Chen et al. 1995. BIOCHEM: Pharmacol. 50: 1257-1264; Creech Kraft et al. 1994. BIOCHEM: J. 301: 111-119; Lanvers et al. 1996. J. Chromatogr. B Biomed. Appl. 685: 233-240; Maden et al. 1998. DEVELOPMENT: 125: 4133-4144; Wagner et al. 1992. DEVELOPMENT: 116: 55-66). This paper describes an HPLC/mass spectrometry/mass spectrometry product ion scan (HPLC/MS(n)) procedure for the analysis of retinoids that employs atmospheric pressure chemical ionization MS. The retinoids are separated by normal-phase column chromatography with a linear hexane isopropanol-dioxane gradient. Each retinoid is detected by a unique series of MS(n) functions set at optimal collision-induced dissociation energy (30% to 32%) for all MS(n) steps. The scan events are divided into three segments, based on HPLC elution order, to maximize the mass spectrometer duty cycle. The all-trans, 9-cis, and 13-cis RA isomers are separated, if desired, by an isocratic hexane dioxane-isopropanol mobile phase. This paper describes an HPLC/MS(n) procedure possessing high sensitivity and specificity for retinoids. PMID- 12091500 TI - Nonenzymatic synthesis of glycerolipids catalyzed by imidazole. AB - Imidazole catalyzed acylations of lysolipids by acyl-CoAs in water at room temperature and at a pH close to neutrality. In the presence of oleoyl-CoA and either lysophosphatidylcholine, 1-palmitoyl-sn-glycero-3-phosphocholine (LPC); lysophosphatidylglycerol, monoacyl-sn-glycero-3-phosphoglycerol; lysophosphatidyl acid, 1-oleoyl-sn-glycero-3-phosphate; lysophosphatidylserine, monoacyl-sn glycero-3-phosphoserin; or lysophosphatidylethanolamine, monoacyl-sn-glycero-3 phosphoethanolamine, the corresponding phospholipids were synthesized. Similarly, the use of lyso-platelet activating factor, an ether analog of LPC, yielded the formation of 1-O-alkyl-2-oleoyl-sn-glycero-3-phosphocholine. In the presence of LPC, an imidazole-catalyzed synthesis of phosphatidylcholine (PC) occurred when medium, long, and very long chain acyl-CoAs were added. With hydroxyacyl-CoA, a similar PC synthesis was obtained. The process described in the present paper appears to offer several potential applications of interest for the synthesis of glycerophospholipids and triglycerides with labeled and/or an unusual or fragile fatty acid, or when suitable acyltransferases have not yet been described in the literature and/or are not commercially available. The method described is very safe and simple since lipids can be synthesized in tubes containing 0.7% imidazole in water, and left for a few hours at room temperature on the bench. PMID- 12091501 TI - Analysis and quantitation of biotinylated apoB-containing lipoproteins with streptavidin-Cy3. AB - Non denaturing gradient gel electrophoresis (GGE) is commonly used to analyze the size distribution of lipoprotein particles. Its relatively low sensitivity and linear dynamic range limit use of GGE to quantify protein content of lipoproteins. We demonstrate a new high sensitivity method for analysis and quantitation of biotinylated apolipoprotein B (apoB)-containing lipoproteins using a fluorescent streptavidin-Cy3 conjugate and non covalent preelectrophoretic binding. Forty-four lipoprotein subfractions spanning the VLDL and LDL particle spectrum subfractions (11 each from four human subjects) were prepared by density gradient ultracentrifugation. An aliquot of each sample was biotinylated and GGE was performed. Gels also were stained for lipid with Oil Red O (32 samples) and for protein with Coomassie Brilliant Blue (30 samples). There was a significant relationship between the Cy3 fluorescent label area under the curve and the mass of apoB (P < 0.02-0.004) and total cholesterol (P < 0.03 0.004). Particle diameters of each absorbence/fluorescent peak were comparable between Oil-Red O and streptavidin-Cy3 treated biotinylated lipoproteins (+/-3.54 A, P = 0.3). Biotinylation and prestaining of lipoprotein particle with streptavidin-Cy3 provides a new fluorescence-based method for detection and quantitative analysis of lipoprotein subspecies by gradient gel electrophoresis. PMID- 12091502 TI - The determination of gases in blood and other solutions by vacuum extraction and manometric measurement. I. PMID- 12091503 TI - Management of papillary and follicular thyroid cancer. PMID- 12091504 TI - Drug-facilitated sexual assault, 'ladettes' and alcohol. PMID- 12091505 TI - The autopsy: lessons from the National Confidential Enquiry into Perioperative Deaths. PMID- 12091506 TI - Specialist nurses in gastroenterology. PMID- 12091507 TI - Antiphospholipid syndrome. PMID- 12091508 TI - Effect of breaking bad news on patients' perceptions of doctors. AB - The breaking of bad news is a routine but difficult task for many health professionals. There are numerous anecdotes of insensitive practice but the subject has attracted little systematic research. We therefore interviewed 106 patients with advanced cancer (from an original sample of 195) to assess their perceptions of the doctors involved in their care. Aspects of the 'breaking bad news' event were recorded during discussion of the illness history and were subsequently rated. Participants were also asked to nominate doctors under the headings 'most helpful' and 'less helpful', and completed standardized psychological screening questionnaires. In 94 of the 106 cases the bad news had been given by a doctor, usually a surgeon. Of the 13 doctors categorized as 'most helpful' when breaking bad news, 8 were general practitioners; of the 7 categorized as 'less helpful' all were surgeons. 69% of patients were neutral or positive about the bad-news consultation, but 20% were negative and 6% very negative. Doctors in surgical specialties were significantly more likely to be rated poorly than non-surgical specialists or general practitioners. Surgeons were the group of doctors most likely to break bad news, but non-surgical doctors were rated more positively in performance of the task. This finding has implications for training. PMID- 12091509 TI - Wet combing for head lice: feasibility in mass screening, treatment preference and outcome. AB - There is no scientific consensus on the best way to control head louse infestation in schoolchildren. A study was conducted to test the feasibility and acceptability of a screening campaign by wet combing and a community approach to head-louse control with home visits, and to explore parents' treatment preferences and treatment outcomes. A non-controlled intervention (advice on treatment options offered to all positive children) was nested within an epidemiological prevalence study. All children in three primary schools in Ghent, Belgium, were invited to take part in screening by wet combing (n=677, 3-11 years). Positive children were offered structural treatment advice, a home visit on day 7, and a check by wet combing on day 14. 83% of the children were screened. The prevalence of active infestation (living moving lice) was 13.0% in school 1 and 19.5% in school 3. In school 2, prevalence of signs of active and past infestation was 40.7%. A home visit was made to 58% of the positive children. 85% of the positive children were screened again on day 14. Wet combing was the most widely used treatment, followed by chemical treatment and a combination of the two. In school 1 and 3 51% were cured, and in school 2 24% became nit-free. A wet combing screening campaign and a community-oriented approach to head-louse control is feasible though resource-intensive. The prevalence of head lice was high and the cure rate was low, with either topical treatments or wet combing. PMID- 12091510 TI - Pacemaker explosions in crematoria: problems and possible solutions. AB - The number of artificial cardiac pacemakers is increasing, as is the number of bodies being cremated. Because of the explosive potential of pacemakers when heated, a statutory question on the cremation form asks whether the deceased has a pacemaker and if so whether it has been removed. We sent a questionnaire to all the crematoria in the UK enquiring about the frequency, consequences and prevention of pacemaker explosions. We found that about half of all crematoria in the UK experience pacemaker explosions, that pacemaker explosions may cause structural damage and injury and that most crematoria staff are unaware of the explosive potential of implantable cardiac defibrillators. Crematoria staff rely on the accurate completion of cremation forms, and doctors who sign cremation forms have a legal obligation to provide such information. PMID- 12091512 TI - Sudden visual loss in asthma. PMID- 12091511 TI - Delay in diagnosis of slipped upper femoral epiphysis. AB - Treatment of slipped upper femoral epiphysis (SUFE) is directed at preventing progressive slippage, minimizing deformity and avoiding avascular necrosis and chondrolysis. Delay in treatment adversely affects long-term outcomes. In a retrospective study we assessed delays between symptom onset and evaluation of the patient in an orthopaedic department. 27 patients aged 10-16 years were grouped by source of referral (general practitioner or accident and emergency department), and hips were classified as stable or unstable according to ability to bear weight. The 27 children had 37 affected hips, 31 stable and 6 unstable. In the 20 patients referred by general practitioners, mean delay from symptom onset to orthopaedic evaluation was 119 days (range 2-504); in the 7 referred from accident and emergency departments it was 95 days (1-482). In the latter group the slips were more likely to be acute and unstable. 9 (45%) of the patients in the general-practitioner group had hip radiography before referral, all correctly diagnosed though not all the examinations included the recommended frog-lateral views. Long delays between onset and diagnosis of SUFE are most likely in patients with mild symptoms, able to bear weight on the hip. Any adolescent with undiagnosed hip or knee pain that has lasted more than a week should undergo radiological investigation of the hip, with frog-lateral as well as anteroposterior views. PMID- 12091513 TI - Rapid enlargement of a bone cyst after biopsy. PMID- 12091514 TI - Recurrent megajejunum in an adult. PMID- 12091515 TI - Neuromuscular blocking drugs: discovery and development. PMID- 12091516 TI - An unlikely aphasiologist: D J Larrey (1766-1842). PMID- 12091517 TI - Research on complementary medicine. PMID- 12091518 TI - Decline in rehabilitation services. PMID- 12091519 TI - Research on complementary medicine. PMID- 12091520 TI - Missed outpatient appointments. PMID- 12091521 TI - The death of Claudius. PMID- 12091523 TI - Public health in the NHS. PMID- 12091522 TI - Waiting in the NHS. PMID- 12091524 TI - Revalidation of the retired. PMID- 12091527 TI - Foundations for the future. PMID- 12091528 TI - Synaptic activity in chronically injured, epileptogenic sensory-motor neocortex. AB - We recorded spontaneous and evoked synaptic currents in pyramidal neurons of layer V in chronically injured, epileptogenic neocortex to assess changes in the efficacy of excitatory and inhibitory neurotransmission that might promote cortical hyperexcitability. Partial sensory-motor neocortical isolations with intact blood supply ("undercuts") were made in 20 rats on postnatal day 21-25 and examined 2-6 wk later in standard brain slice preparations using whole cell patch clamp techniques. Age-matched, uninjured naive rats (n = 20) were used as controls. Spontaneous and miniature excitatory and inhibitory postsynaptic currents (s- and mEPSCs; s- and mIPSCs) were recorded using patch-clamp techniques. The average frequency of s- and mEPSCs was significantly higher, while that of s- and mIPSCs was significantly lower in neurons of undercuts versus controls. The increased frequency of excitatory events was due to an increase in both s- and mEPSC frequency, suggesting an increased number of excitatory contacts and/or increased release probability at excitatory terminals. No significant difference was observed in 10-90% rise time of these events. The input-output slopes of fast, short-latency, alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid/kainate (AMPA/KA) receptor-mediated components of evoked EPSCs were steeper in undercuts than in controls. The peak amplitude of the AMPA/KA component of EPSCs evoked by supra-threshold stimuli was significantly greater in the partially isolated neocortex. In contrast, the N methyl-D-aspartate receptor-mediated component of evoked EPSCs was not significantly different in neurons of injured versus control cortex, suggesting that the increased AMPA/KA component was due to postsynaptic alterations. Results support the conclusion that layer V pyramidal neurons receive increased AMPA/KA receptor-mediated excitatory synaptic drive and decreased GABA(A) receptor mediated inhibition in this chronically injured, epileptogenic cortex. This shift in the balance of excitatory and inhibitory synaptic activation of layer V pyramidal cells toward excitation might be maladaptive and play a critical role in epileptogenesis. PMID- 12091529 TI - Vestibuloocular reflex dynamics during high-frequency and high-acceleration rotations of the head on body in rhesus monkey. AB - For frequencies >10 Hz, the vestibuloocular reflex (VOR) has been primarily investigated during passive rotations of the head on the body in humans. These prior studies suggest that eye movements lag head movements, as predicted by a 7 ms delay in the VOR reflex pathways. However, Minor and colleagues recently applied whole-body rotations of frequencies < or =15 Hz in monkeys and found that eye movements were nearly in phase with head motion across all frequencies. The goal of the present study was to determine whether VOR response dynamics actually differ significantly for whole-body versus head-on-body rotations. To address this question, we evaluated the gain and phase of the VOR induced by high frequency oscillations of the head on the body in monkeys by directly measuring both head and eye movements using the magnetic search coil technique. A torque motor was used to rotate the heads of three Rhesus monkeys over the frequency range 5-25 Hz. Peak head velocity was held constant, first at +/-50 degrees /s and then +/-100 degrees /s. The VOR was found to be essentially compensatory across all frequencies; gains were near unity (1.1 at 5 Hz vs. 1.2 at 25 Hz), and phase lag increased only slightly with frequency (from 2 degrees at 5 Hz to 11 degrees at 25 Hz, a marked contrast to the 63 degrees lag at 25 Hz predicted by a 7-ms VOR latency). Furthermore, VOR response dynamics were comparable in darkness and when viewing a target and did not vary with peak velocity. Although monkeys offered less resistance to the initial cycles of applied head motion, the gain and phase of the VOR did not vary for early versus late cycles, suggesting that an efference copy of the motor command to the neck musculature did not alter VOR response dynamics. In addition, VOR dynamics were also probed by applying transient head perturbations with much greater accelerations (peak acceleration >15,000 degrees /s(2)) than have been previously employed. The VOR latency was between 5 and 6 ms, and mean gain was close to unity for two of the three animals tested. A simple linear model well described the VOR responses elicited by sinusoidal and transient head on body rotations. We conclude that the VOR is compensatory over a wide frequency range in monkeys and has similar response dynamics during passive rotation of the head on body as during passive rotation of the whole body in space. PMID- 12091530 TI - AMPA receptor-mediated miniature synaptic calcium transients in GluR2 null mice. AB - AMPA-type glutamate receptors are normally Ca(2+) impermeable due to the expression of the GluR2 receptor subunit. By using GluR2 null mice we were able to detect miniature synaptic Ca(2+) transients (MSCTs) associated with AMPA-type receptor-mediated miniature synaptic currents at single synapses in primary cortical cultures. MSCTs and associated Ca(2+) transients were monitored under conditions that isolated responses mediated by AMPA or N-methyl-D-aspartate (NMDA) receptors. As expected, addition of the antagonist 6-cyano-7 nitroquinoxalene-2,3-dione (CNQX, 3 microM) blocked the AMPA receptor-mediated MSCTs. Voltage-gated Ca(2+) channels did not contribute to AMPA MSCTs because CdCl(2) (0.1-0.2 mM) did not significantly alter the frequency or the amplitude of the MSCTs. The amplitude of AMPA MSCTs appeared to be regulated independently from event frequency since the two measures were not correlated (R = 0.023). Synapses were identified that only expressed MSCTs attributed to either NMDA or AMPA receptors. At synapses with only NMDA responses, MSCT amplitude was significantly lower (by 40%) than synapses expressing both NMDA and AMPA responses. At synapses that showed MSCTs mediated by both AMPA and NMDA receptors, the amplitude of the transients in each condition was positively correlated (R = 0.94). Our results suggest that when AMPA and NMDA receptors are co-expressed at synapses, mechanisms exist to ensure proportional scaling of each receptor type that are distinct from the presynaptic factors controlling the frequency of miniature release. PMID- 12091531 TI - Functional characterization of kainate receptors in the rat nucleus accumbens core region. AB - The nucleus accumbens, a brain region involved in motivation, attention, and reward, receives substantial glutamatergic innervation from many limbic structures. This excitatory glutamatergic input plays an integral role in both normal and pathophysiological states. Despite the importance of glutamatergic transmission in the nucleus accumbens, the specific receptor subtypes that mediate glutamatergic signaling in this brain region have not been fully characterized. The current study sought to examine the possible role of the kainate subclass of glutamate receptor in the nucleus accumbens. Kainate receptors are relatively poorly understood members of the ionotropic glutamate receptor family and are highly expressed in the nucleus accumbens. Recent studies have highlighted a number of novel pre- and postsynaptic functions of kainate receptors in several other brain regions. Using the whole cell patch-clamp technique, we report the first demonstration of functional kainate receptors on neurons within the core region of the nucleus accumbens. In addition, we present evidence that activation of kainate receptors in this brain region inhibits excitatory synaptic transmission via a presynaptic mechanism. PMID- 12091532 TI - Glutamate mediates an excitatory influence of the paraventricular hypothalamic nucleus on the dorsal motor nucleus of the vagus. AB - Data have shown that the paraventricular nucleus of the hypothalamus (PVN) and the dorsal motor nucleus of the vagus (DMNV) play important roles in the regulation of gastrointestinal function and eating behavior. Anatomical studies have demonstrated direct projections from the PVN to the DMNV and physiological studies showed that the DMNV mediates many of the effects of PVN stimulation and electrical current stimulation of the PVN excites a subset of DMNV neurons. The aim of this study was to characterize the role of glutamate receptors in the excitatory influence of the PVN on gut-related DMNV neurons. Using single-cell recording techniques, we determined the effects of kynurenic acid, 6-cyano-7 nitroquinoxalene-2,3-dione (CNQX), and DL-2-amino-5-phosphonopentanoic acid (DL AP5) on the increase in firing rate due to electrical current stimulation of the PVN. In initial experiments, we studied 24 DMNV neurons excited by electrical current stimulation of the PVN. Kynurenic acid, a broad-spectrum glutamate receptor antagonist, prevented the PVN effect in 22 neurons and significantly attenuated the effect in the other cells. Nine of these neurons demonstrated an inhibition in firing rate with PVN stimulation after pretreatment with kynurenic acid. In a separate group of 12 neurons, we determined the effects of CNQX (1.2 nmol) injected into the DMNV. This AMPA receptor antagonist completely blocked the excitatory response to PVN stimulation of six DMNV neurons and significantly attenuated the response of the other six DMNV neurons. The addition of 1.2 nmol DL-AP5, a N-methyl-D-aspartate (NMDA) receptor antagonist, further attenuated the response to PVN stimulation in four of the five DMNV neurons that were still excited after CNQX treatment. The fifth neuron demonstrated PVN- induced inhibition of firing rate after treatment with CNQX and DL-AP5. In a separate group of 11 DMNV neurons excited by electrical stimulation of the PVN, DL-AP5 partially attenuated the excitatory responses of only four DMNV neurons and did not block the excitation of any cells. The mean latency (14 neurons tested) from the PVN to the DMNV was 37.71 +/- 2.40 (SE) ms. Monosynaptic action potentials and excitatory postsynaptic potentials were demonstrated in three DMNV neurons by intracellular recording. Our results indicate that glutamate released from PVN neurons projecting to the DMNV excite the gut-related vagal motor neurons by acting predominantly on the AMPA receptor. The NMDA receptor plays only a minor role in the excitatory effect. PMID- 12091533 TI - Inhibition of backpropagating action potentials in mitral cell secondary dendrites. AB - The mammalian olfactory bulb is a geometrically organized signal-processing array that utilizes lateral inhibitory circuits to transform spatially patterned inputs. A major part of the lateral circuitry consists of extensively radiating secondary dendrites of mitral cells. These dendrites are bidirectional cables: they convey granule cell inhibitory input to the mitral soma, and they conduct backpropagating action potentials that trigger glutamate release at dendrodendritic synapses. This study examined how mitral cell firing is affected by inhibitory inputs at different distances along the secondary dendrite and what happens to backpropagating action potentials when they encounter inhibition. These are key questions for understanding the range and spatial dependence of lateral signaling between mitral cells. Backpropagating action potentials were monitored in vitro by simultaneous somatic and dendritic whole cell recording from individual mitral cells in rat olfactory bulb slices, and inhibition was applied focally to dendrites by laser flash photolysis of caged GABA (2.5-microm spot). Photolysis was calibrated to activate conductances similar in magnitude to GABA(A)-mediated inhibition from granule cell spines. Under somatic voltage-clamp with CsCl dialysis, uncaging GABA onto the soma, axon initial segment, primary and secondary dendrites evoked bicuculline-sensitive currents (up to -1.4 nA at 60 mV; reversal at approximatety 0 mV). The currents exhibited a patchy distribution along the axon and dendrites. In current-clamp recordings, repetitive firing driven by somatic current injection was blocked by uncaging GABA on the secondary dendrite approximately 140 microm from the soma, and the blocking distance decreased with increasing current. In the secondary dendrites, backpropagated action potentials were measured 93-152 microm from the soma, where they were attenuated by a factor of 0.75 +/- 0.07 (mean +/- SD) and slightly broadened (1.19 +/- 0.10), independent of activity (35-107 Hz). Uncaging GABA on the distal dendrite had little effect on somatic spikes but attenuated backpropagating action potentials by a factor of 0.68 +/- 0.15 (0.45-0.60 microJ flash with 1-mM caged GABA); attenuation was localized to a zone of width 16.3 +/ 4.2 microm around the point of GABA release. These results reveal the contrasting actions of inhibition at different locations along the dendrite: proximal inhibition blocks firing by shunting somatic current, whereas distal inhibition can impose spatial patterns of dendrodendritic transmission by locally attenuating backpropagating action potentials. The secondary dendrites are designed with a high safety factor for backpropagation, to facilitate reliable transmission of the outgoing spike-coded data stream, in parallel with the integration of inhibitory inputs. PMID- 12091534 TI - Effects of excitatory modulation on intrinsic properties of turtle motoneurons. AB - The purpose of this study was to quantify the effects of excitatory modulation on the intrinsic properties of motoneurons (MNs) in slices of spinal cord taken from the adult turtle. Responses were noted following application of an excitatory modulator: serotonin (5-HT), muscarine, trans-1-amino-1,3-cyclopentane dicarboxylic acid (tACPD), or all three combined. A sample of 44 MNs was divided into 2 groups, on the basis of whether MNs did (28/44) or did not (16/44) demonstrate a nifedipine-sensitive acceleration of discharge during a 2-s, intracellularly injected stimulus pulse. Such acceleration indicates the development of a plateau potential (PP). Excitatory modulation lowered the MNs' resting potential, increased input resistance, decreased rheobase, reduced several afterhyperpolarization values, and shifted the conventional, one-phase stimulus current-spike frequency (I-f) relation to the left. For both MN groups, the relative efficacy of excitatory modulation on both non-PP and PP MNs was generally in the following order: combined application > 5-HT approximately muscarine > tACPD. In many instances, the effects of modulation differed significantly for non-PP versus PP MNs, the most pronounced being in their I-f relation. To describe this difference, it was necessary to measure a two-phase relation. In PP MNs, excitatory modulation considerably increased the slope of the first (initial) phase and flattened the second (later) phase of this relation. The latter result bore similarities to that obtained in a previous study, which addressed MN firing behavior during fictive locomotion of the high decerebrate cat. PMID- 12091535 TI - Roles of ATP in depletion and replenishment of the releasable pool of synaptic vesicles. AB - Synaptic terminals of retinal bipolar neurons contain a pool of readily releasable synaptic vesicles that undergo rapid calcium-dependent release. ATP hydrolysis is required for the functional refilling of this vesicle pool. However, it was unclear which steps required ATP hydrolysis: delivery of vesicles to their anatomical release sites or preparation of synaptic vesicles and/or the secretory apparatus for fusion. To address this, we dialyzed single synaptic terminals with ATP or the poorly hydrolyzable analogue ATP-gammaS and examined the size of the releasable pool, refilling of the releasable pool, and the number of vesicles at anatomical active zones. After minutes of dialysis with ATP gammaS, vesicles already in the releasable pool could still be discharged. This pool was not functionally refilled despite the fact that its anatomical correlate, the number of synaptic vesicles tethered to active zone synaptic ribbons, was completely normal. We conclude 1) because the existing releasable pool is stable during prolonged inhibition of ATP hydrolysis, whereas entry into the functional pool is blocked, a vesicle on entering the pool will tend to remain there until it fuses; 2) because the anatomical pool is unaffected by inhibition of ATP hydrolysis, failure to refill the functional pool is not caused by failure of vesicle movement; 3) local vesicle movements important for pool refilling and fusion are independent of conventional ATP-dependent motor proteins; and 4) ATP hydrolysis is required for the biochemical transition of vesicles and/or release sites to fusion-competent status. PMID- 12091536 TI - Group I mGluRs increase excitability of hippocampal CA1 pyramidal neurons by a PLC-independent mechanism. AB - Previous studies have implicated phospholipase C (PLC)-linked Group I metabotropic glutamate receptors (mGluRs) in regulating the excitability of hippocampal CA1 pyramidal neurons. We used intracellular recordings from rat hippocampal slices and specific antagonists to examine in more detail the mGluR receptor subtypes and signal transduction mechanisms underlying this effect. Application of the Group I mGluR agonist (RS)-3,5-dihydroxyphenylglycine (DHPG) suppressed slow- and medium-duration afterhyperpolarizations (s- and mAHP) and caused a consequent increase in cell excitability as well as a depolarization of the membrane and an increase in input resistance. Interestingly, with the exception of the suppression of the mAHP, these effects were persistent, and in the case of the sAHP lasting for more than 1 h of drug washout. Preincubation with the specific mGluR5 antagonist, 2-methyl-6-(phenylethynyl)-pyridine (MPEP), reduced but did not completely prevent the effects of DHPG. However, preincubation with both MPEP and the mGluR1 antagonist LY367385 completely prevented the DHPG-induced changes. These results demonstrate that the DHPG induced changes are mediated partly by mGluR5 and partly by mGluR1. Because Group I mGluRs are linked to PLC via G-protein activation, we also investigated pathways downstream of PLC activation, using chelerythrine and cyclopiazonic acid to block protein kinase C (PKC) and inositol 1,4,5-trisphosphate-(IP(3)) activated Ca(2+) stores, respectively. Neither inhibitor affected the DHPG induced suppression of the sAHP or the increase in excitability nor did an inhibitor of PLC itself, U-73122. Taken together, these results argue that in CA1 pyramidal cells in the adult rat, DHPG activates mGluRs of both the mGluR5 and mGluR1 subtypes, causing a long-lasting suppression of the sAHP and a consequent persistent increase in excitability via a PLC-, PKC-, and IP(3)-independent transduction pathway. PMID- 12091537 TI - Posthatching locomotor experience alters locomotor development in chicks. AB - We have previously demonstrated that, even though chicks are very precocial and can locomote within hours of hatching, they require a period of time to develop a mature stable walk. As an example, 1- to 2-day-old animals move with disproportionately small stride lengths compared with 10- to 14-day-old animals. The purpose of this study was to determine whether the maturation of walking, including the development of a mature stride length, depends on locomotor experience. We also investigated the development and experience-dependence nature of head bobbing, an optokinetic behavior that occurs during walking in birds. Chicks were randomly assigned to one of three groups receiving either increased locomotor experience (i.e., treadmill exercise), decreased locomotor experience (i.e., decreased housing space), or no alteration in locomotor experience. To assess the dependence of locomotor maturation on N-methyl-D-aspartate (NMDA)-type glutamate receptors, animals in each group were either given an NMDA antagonist (MK-801, 1 mg/kg intramuscularly daily) or saline control. Locomotor characteristics (stride length, leg support durations, horizontal head excursions) were quantified from videotaped recordings of chicks walking overground unrestrained on posthatching days 1, 2, 4, 6, 8, and 10. Animals subject to exercise restriction for at least 6 days moved with shortened stride lengths compared with age-matched treadmill-exercised or control animals, a change that was maintained for the duration of the study. NMDA antagonism also resulted in shortened stride lengths. Head bobbing behavior matured during the same posthatching time period. The rate of this maturation was also decreased by exercise restriction. Thus locomotor experience is required for normal development of locomotor behavior, even in very precocial animals. These results are discussed in terms of the possible neuroanatomical and neurophysiological mechanisms underlying experience- and activity-dependent changes during motor development. PMID- 12091538 TI - Responses of ankle extensor and flexor motoneurons to transcranial magnetic stimulation. AB - Transcranial magnetic stimulation (TMS) of the motor cortex excites limb muscles of the contralateral side of the body. Reports of poorly defined, or a complete lack of systematic excitatory responses of soleus motoneurons compared with those of tibialis anterior (TA) motoneurons has led to the proposal that while all ankle flexor motoneurons receive strong corticomotoneuronal connections, very few soleus motoneurons do. In addition, the connections to these few motoneurons are weak. The nature of corticomotoneuronal connections onto these two motoneuron pools was re-evaluated in the following experiments. The leg area of the left motor cortex was stimulated with a large double-cone coil using Magstim 200, while surface electromyographic (EMG) and single motor unit (SMU) responses were recorded from soleus and TA muscles of healthy adult subjects. Under resting conditions, the onset (25-30 ms) and duration of concomitantly recorded short latency motor evoked potentials (MEPs) in surface EMG from both muscles were similar. The input-output relationships of the simultaneously recorded soleus and TA EMG responses showed much greater increases in TA MEPs compared with soleus MEPs with identical increases in stimulus intensity. Under resting and nonisometric conditions, a later peak with onset latency of approximately 100 ms was observed in soleus. During isometric conditions or with vibration of the TA tendon, the second soleus peak was abolished indicating reflex origin of this peak. Recordings from 42 soleus and 39 TA motor units showed clear response peaks in the peristimulus time histograms (PSTHs) of every unit. Two statistical tests were done to determine the onset and duration of response peaks in the PSTHs. With chi(2) test, the duration was 6.9 +/- 4.2 ms (mean +/- SD) for soleus and 5.1 +/- 2.1 ms for TA. Using the criterion of discerning a peak by bin counts being three SDs above background, the duration was 10.0 +/- 4.4 ms for soleus and 7.8 +/- 2.6 ms for TA. Results of these experiments do not suggest a lack of systematic corticomotoneuronal connections on soleus motoneurons when compared with those on TA, though some differences in the strengths of corticomotoneuronal connections onto the two pools do exist. PMID- 12091540 TI - Differential modulation of nucleus accumbens synapses. AB - The nucleus accumbens (NAcc) is a brain region involved in functions ranging from motivation and reward to feeding and drug addiction. The NAcc is typically divided into two major subdivisions, the shell and the core. The primary output neurons of both of these areas are medium spiny neurons (MSNs), which are quiescent at rest and depend on the relative input of excitatory and inhibitory synapses to determine when they fire action potentials. These synaptic inputs are, in turn, regulated by a number of neurochemical signaling agents that can ultimately influence information processing in the NAcc. The present study characterized the ability of three major signaling pathways to modulate synaptic transmission in NAcc MSNs and compared this modulation across different synapses within the NAcc. The opioid [Met](5)enkephalin (ME) inhibited excitatory postsynaptic currents (EPSCs) in shell MSNs, an effect mediated primarily by micro-opioid receptors. Forskolin, an activator of adenylyl cyclase, potentiated shell EPSCs. An analysis of miniature EPSCs indicated a primarily presynaptic site of action, although a smaller postsynaptic effect may have also contributed to the potentiation. Adenosine and an adenosine A(1)-receptor agonist inhibited shell EPSCs, although no significant tonic inhibition by endogenous adenosine was detected. The effects of these signaling agents were then compared across four different synapses in the NAcc: glutamatergic EPSCs and GABAergic inhibitory postsynaptic currents (IPSCs) in both the core and shell subregions. ME inhibited all four of these synapses but produced a significantly greater inhibition of shell IPSCs than the other synapses. Forskolin produced an increase in transmission at each of the synapses tested. However, analysis of miniature IPSCs in the shell showed no sign of a postsynaptic contribution to this potentiation, in contrast to the shell miniature EPSCs. Tonic inhibition of synaptic currents by endogenous adenosine, which was not observed in shell EPSCs, was clearly present at the other three synapses tested. These results indicate that neuromodulation can vary between the different subregions of the NAcc and between the different synapses within each subregion. This may reflect differences in neuronal interconnections and functional roles between subregions and may contribute to the effects of drugs acting on these systems. PMID- 12091539 TI - Acid-activated cation currents in rat vallate taste receptor cells. AB - Sour taste is mediated by acids with the degree of sourness a function of proton concentration. Recently, several members of the acid-sensing ion channel subfamily (ASICs) were cloned from taste cells and proposed to mediate sour taste. However, it is not known whether sour responses in taste cells resemble the responses mediated by ASICs. Using the whole cell patch-clamp technique and Na(+) imaging, we have characterized responses to acid stimuli in isolated rat vallate taste cells. Citric acid (pH 5) induced a large, rapidly activating inward current in most taste cells tested. The response showed various degrees of desensitization with prolonged stimulation. Current amplitudes were pH dependent, and adapting with acidic bath solutions reduced subsequent responses to acid stimulation. Amiloride (100-500 microM) partially and reversibly suppressed the acid-induced current. The current-voltage relationship showed reversal potential near the Na(+) equilibrium potential, suggesting that the current is carried predominantly by Na(+). These data were consistent with Na(+) imaging experiments showing that acid stimulation resulted in increases in intracellular Na(+). Taken together, these data indicate that acid-induced currents in vallate taste cells share general properties with ASICs expressed in heterologous cells and sensory neurons that express ASIC subunits. The large amplitude of the current and its existence in a high percentage of taste cells imply that ASICs or ASIC-like channels may play a prominent role in sour-taste transduction. PMID- 12091541 TI - Intrinsic neuronal properties of the chick nucleus angularis. AB - In vitro whole cell recording revealed intrinsic firing properties and single cell morphology in the cochlear nucleus angularis (NA) of the chick. We classified three major classes of neurons: one-spike, damped, and tonic. A delayed inward rectifying current was observed in all classes during hyperpolarization injections. One-spike neurons responded with a single spike to depolarizing current injection and had small (stubby) radiate dendritic trees. Damped neurons responded with only a few spikes at the onset of positive current injection. More positive current inputs led to a damped response. Damped cell dendrites had a planar orientation parallel to the isofrequency axis in NA. Tonic cells produced trains of action potentials in response to a depolarizing current injection. Three variations of the tonic type had multipolar morphology, with dendrites oriented either radially (I and III) or perpendicular to the tonotopic axis (II; vertical). Tonics I and III differed in the shape of their action potential undershoot. Thus NA is both physiologically and morphologically heterogeneous. PMID- 12091542 TI - Sensory integration in presynaptic inhibitory pathways during fictive locomotion in the cat. AB - The aim of this study is to understand how sensory inputs of different modalities are integrated into spinal cord pathways controlling presynaptic inhibition during locomotion. Primary afferent depolarization (PAD), an estimate of presynaptic inhibition, was recorded intra-axonally in group I afferents (n = 31) from seven hindlimb muscles in L(6)-S(1) segments during fictive locomotion in the decerebrate cat. PADs were evoked by stimulating alternatively low-threshold afferents from a flexor nerve, a cutaneous nerve and a combination of both. The fictive step cycle was divided in five bins and PADs were averaged in each bin and their amplitude compared. PADs evoked by muscle stimuli alone showed a significant phase-dependent modulation in 20/31 group I afferents. In 12/20 afferents, the cutaneous stimuli alone evoked a phase-dependent modulation of primary afferent hyperpolarization (PAH, n = 9) or of PADs (n = 3). Combining the two sensory modalities showed that cutaneous volleys could significantly modify the amplitude of PADs evoked by muscle stimuli in at least one part (bin) of the step cycle in 17/31 (55%) of group I afferents. The most common effect (13/17) was a decrease in the PAD amplitude by 35% on average, whereas it was increased by 17% on average in the others (4/17). Moreover, in 8/13 afferents, the PAD reduction was obtained in 4/5 bins i.e., for most of the duration of the step cycle. These effects were seen in group I afferents from all seven muscles. On the other hand, we found that different cutaneous nerves had quite different efficacy; the superficial peroneal (SP) being the most efficient (85% of trials) followed by Saphenous (60%) and caudal sural (44%) nerves. The results indicate that cutaneous interneurons may act, in part, by modulating the transmission in PAD pathways activated by group I muscle afferents. We conclude that cutaneous input, especially from the skin area on the dorsum of the paw (SP), could subtract presynaptic inhibition in some group I afferents during perturbations of stepping (e.g., hitting an obstacle) and could thus adjust the influence of proprioceptive feedback onto motoneuronal excitability. PMID- 12091543 TI - Directional selectivity for FM sweeps in the suprageniculate nucleus of the mustached bat medial geniculate body. AB - Mustached bats emit echolocation and communication calls containing both constant frequency (CF) and frequency-modulated (FM) components. Previously we found that 86% of neurons in the ventral division of the external nucleus of the inferior colliculus (ICXv) were directionally selective for linear FM sweeps and that selectivity was dependent on sweep rate. The ICXv projects to the suprageniculate nucleus (Sg) of the medial geniculate body. In this study, we isolated 37 single units in the Sg and measured their responses to best excitatory frequency (BEF) tones and linear 12-kHz upward and downward FM sweeps centered on the BEF. Sweeps were presented at durations of 30, 12, and 4 ms, yielding modulation rates of 400, 1,000, and 3,000 kHz/s. Spike count versus level functions were obtained at each modulation rate and compared with BEF controls. Sg units responded well to both tones and FM sweeps. BEFs clustered at 58 kHz, corresponding to the dominant CF component of the sonar signal. Spike count functions for both tones and sweeps were predominantly non-monotonic. FM directional selectivity was significant in 53-78% of the units, depending on modulation rate and level. Units were classified as up-selective (52%), down-selective (24%), or bi-directional (non selective, 16%); a few units (8%) showed preferences that were either rate- or level-dependent. Most units showed consistent directional preferences at all SPLs and modulation rates tested, but typically showed stronger selectivity at lower sweep rates. Directional preferences were attributable to suppression of activity by sweeps in the non-preferred direction (~80% of units) and/or facilitation by sweeps in the preferred direction (~20-30%). Latencies for BEF tones ranged from 4.9 to 25.7 ms. Latencies for FM sweeps typically varied linearly with sweep duration. Most FM latency-duration functions had slopes ranging from 0.4 to 0.6, suggesting that the responses were triggered by the BEF. Latencies for BEF tones and FM sweeps were significantly correlated in most Sg units, i.e., the response to FM was temporally related to the occurrence of the BEF in the FM sweep. FM latency declined relative to BEF latency as modulation rate increased, suggesting that at higher rates response is triggered by frequencies in the sweep preceding the BEF. We conclude that Sg and ICXv units have similar, though not identical, response properties. Sg units are predominantly upsweep selective and could respond to either or both the CF and FM components in biosonar signals in a number of echolocation scenarios, as well as to a variety of communication sounds. PMID- 12091544 TI - Dissociated retinal neurons form periodically active synaptic circuits. AB - Throughout the developing nervous system, immature circuits generate rhythmic activity patterns that influence the formation of adult networks. The cellular mechanisms underlying this spontaneous, correlated activity can be studied in dissociated neuronal cultures. Using calcium imaging and whole cell recording, we showed that cultured dissociated mammalian retinal neurons form networks that produce spontaneous, correlated, highly periodic activity. As the culture matures, the spatial correlations of the periodic calcium transients evolve from being highly synchronized across neighboring cells to propagating across the culture in a wavelike manner reminiscent of retinal waves recorded in vivo. Spontaneous calcium transients and synaptic currents were blocked either by cadmium, tetrodotoxin, or the glutamate receptor antagonist 6,7 dinitroquinoxaline, indicating that the periodic activity was driven primarily by synaptic transmission between retinal ganglion cells. Evoked responses between pairs of ganglion cells exhibited paired-pulse synaptic depression, and the time constant of recovery from this depression was similar to the interval between periodic events. These results suggest that synaptic depression may regulate the frequency of network activity. Together, these findings provide insight into how networks containing primarily excitatory connections generate highly correlated activity. PMID- 12091545 TI - T-Type calcium channel alpha1G and alpha1H subunits in human retinoblastoma cells and their loss after differentiation. AB - Human retinoblastoma cells are multipotent retinal precursor cells capable of differentiating into photoreceptors, neurons, and glia. The current-voltage relation of the undifferentiated cells is dominated by a transient inward current that disappears shortly after differentiation. In 20 mM Ba(2+)-containing bath solutions, the current has an activation midpoint near -25 mV and appears to be fully inactivated at -20 mV. Sr(2+) and Ca(2+) are preferred charge carriers relative to Ba(2+), and the current vanishes in the absence of these divalent cations. Cd(2+) blocks the current with an IC(50) of 160 microM, and Ni(2+) blocks in a biphasic manner with IC(50)s of 22 and 352 microM. The current is unaffected when sodium is replaced with other monovalent cations, and it is insensitive to nifedipine, omega-conotoxin GVIA, omega-agatoxin IVA, and omega conotoxin MVIIC. RT-PCR revealed the presence of alpha 1G and alpha 1H mRNA in undifferentiated cells, but following differentiation, a striking reduction of both alpha 1G and alpha 1H mRNA was found, and this was paralleled by the loss of T-type Ca channel currents. alpha 1I subunit mRNA levels were low in undifferentiated and differentiated cells. These results suggest that T-type Ca channels could play a role in undifferentiated retinoblastoma cell physiology since alpha 1G and alpha 1H Ca channel subunit expression is reduced in cells that have differentiated and exited the cell cycle. PMID- 12091546 TI - Long-term deprivation of substance P in PPT-A mutant mice alters the anoxic response of the isolated respiratory network. AB - The aim of this study was to elucidate the role of the neuromodulator substance P and its related tachykinin neurokinin A (NKA) in the homeostasis of respiratory activity. Respiratory activities, in form of fictive eupneic and sigh activities, were recorded extracellularly from the preBotzinger complex (PBC) in normoxic and anoxic conditions using medullary slice preparations. The effect of a blockade of endogenous substance P was assessed by an acute pharmacological blockade of the receptors with spantide in wild-type animals and by the use of preprotachykinin-A (PPT-A) mutants. These mutants lack from birth the PPT-A gene, which codes for the precursor of substance P and NKA. Spantide treatment reduced frequency (-37%, n = 9) and regularity (twofold) of eupneic-like respiratory activity under normoxic conditions, whereas in PPT-A mutants, eupneic-like activity was under normoxic conditions not significantly different from the wild-type mice (WT). The response to short anoxic episodes (5 min) was characterized in the WT by an increase in respiratory frequencies at the onset of anoxia (ratio anoxic/control frequency = 1.9 +/- 0.2, n = 18). This anoxic ratio was unaltered in the presence of spantide (ratio = 2.3 +/- 0.4, n = 8) but increased in the mutant (ratio = 4.1, n = 15). We conclude that endogenously released substance P is important for the maintenance of regular respiratory activity. Short-term blockade of substance P receptors decreases the frequency and regularity of rhythmic activity. Long term deficiency in substance P leads to compensatory mechanisms that result in an apparently normal respiratory activity under normoxic conditions but a significantly altered response of the respiratory network during anoxia. PMID- 12091547 TI - Effects of protein kinase a activation on the responses of primate spinothalamic tract neurons to mechanical stimuli. AB - Behavioral and anatomical studies by our group have suggested that the protein kinase A (PKA) signal transduction cascade contributes to long-term changes in nociceptive processing at the spinal cord level. In this study, we have examined the effects of activation of the PKA cascade on the responses of spinothalamic tract (STT) neurons to peripheral mechanical stimuli in anesthetized and paralyzed monkeys. PKA in the spinal cord was activated by intra-spinal infusion of forskolin, an activator of adenylate cyclase, by microdialysis. There was a consistent increase in responses to mechanical pressure and pinch stimuli in all STT cells tested when forskolin was administered. Enhanced responses remained at relatively high levels when forskolin had been washed out for 30 min. However, in most STT cells tested (65%), the responses to brushing stimuli were not obviously changed when forskolin was given. Background activity was slightly increased when forskolin was administered. An inactive isomer of forskolin, D-forskolin, did not produce significant effects on cellular activity. The sensitization of STT cells to noxious mechanical stimuli produced by forskolin could be blocked by pretreatment of the spinal cord with the PKA inhibitor, N-[2-((p bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamine (H89). The same dose of H89 did not affect the enhanced responses to mechanical stimuli produced by activation of protein kinase G by intra-spinal infusion of 8-bromo-cGMP, indicating that the effect of forskolin was selective. The present data suggest that activation of PKA can preferentially enhance the responses of STT cells to noxious mechanical stimuli without producing an increase in responses to innocuous brushing stimuli. We speculate that the PKA signal transduction cascade may contribute more to secondary mechanical hyperalgesia than to secondary mechanical allodynia. PMID- 12091548 TI - Manipulating objects with internal degrees of freedom: evidence for model-based control. AB - There is substantial evidence that humans possess an accurate and adaptable internal model of the dynamics of their arm that is utilized by the nervous system for controlling arm movements. However, it is not known if such model based strategies are used for controlling dynamical systems outside the body. The need to predict events in the external world is not restricted to the execution of reaching movements or to the handling of rigid tools. Model-based control may also be critical for performing functional tasks with non-rigid objects such as stabilizing a cup of coffee. The present study investigated the strategies used by humans to control simple mass-spring objects. Subjects made straight line reaching movements to a target while interacting with a robotic manipulandum that simulated the dynamics of a one-dimensional mass on a spring. After learning, neither hand nor object kinematics returned to those of free reaching, suggesting that this task was not learned as a perturbation of free reaching. Although there are control strategies (such as slowing the movement of the hand) that would require little or no knowledge of object dynamics, subjects did not adopt these strategies. Instead, they tailored their motor commands to the particular object being manipulated. When object parameters were unexpectedly altered in a way that required no changes in kinematics to successfully complete the task, subjects nonetheless exhibited substantial kinematic deviations. These deviations were consistent with those predicted by a model of the arm-plus-object system driven by a low-impedance controller that incorporated an explicit inverse model of arm plus-object dynamics. The observed behavior could not be reproduced by a controller that relied on modulating hand impedance alone with no inverse model. These results were therefore consistent with the hypothesis that subjects learn to control the kinematics of manipulated objects by forming an internal model that specified the forces to be exerted by the hand on the object to induce the desired motion of that object. PMID- 12091549 TI - Protective effect of high glucose against ischemia-induced synaptic transmission damage in rat hippocampal slices. AB - Cerebral ischemic damage is an important cause of morbidity and mortality. However, there is conflicting evidence regarding the effect of the extracellular glucose concentration in focal and global ischemic injury. This study was designed to investigate this effect in ischemia-induced synaptic transmission damage in rat hippocampal slices. Slices were superfused with artificial cerebrospinal fluid (ACSF) containing various concentrations of glucose before and after ischemia. The evoked somatic postsynaptic population spike (PS) and dendritic field excitatory postsynaptic potential (fEPSP) were extracellularly recorded in the CA1 stratum pyramidal cell layer and s. radiatum after stimulation of the Schaeffer collaterals, respectively. The glucose concentration in ACSF before and after ischemia determined the duration of ischemia tolerated by synaptic transmission as demonstrated by complete recovery of the somatic PS and dendritic fEPSP. Specifically, the somatic PS and dendritic fEPSP completely recovered following 3, 4, and 5 min of ischemia only when slices were superfused with ACSF containing 4, 10, and 20 mM glucose before and after ischemia, respectively. The latencies of the somatic and dendritic ischemic depolarization (ID) occurrence in the CA1 s. pyramidal cell layer and s. radiatum were significantly longer with 10 than 4 mM glucose in ACSF before ischemia and significantly longer with 20 than 10 mM glucose in ACSF before ischemia. Regardless of the glucose concentration in ACSF before and after ischemia, the somatic PS and dendritic fEPSP only partially recovered when ischemia was terminated at the occurrence of ID. These results indicate that high glucose in ACSF during the period before and after ischemia significantly protects CA1 synaptic transmission against in vitro ischemia-induced damage through postponing the occurrence of ID. PMID- 12091550 TI - Different calcium sources are narrowly tuned to the induction of different forms of LTP. AB - The essential role of calcium in the induction of long-term potentiation (LTP) has been well established. In particular, calcium influx via the N-methyl-D aspartate (NMDA) receptor (NMDAR) is important for LTP induction in many pathways. However, the specific roles of other calcium sources in hippocampal LTP are less clear. The aim of the present study was to determine the appropriate conditions and extent to which non-NMDAR Ca(2+) sources contribute to the induction of different forms of LTP in area CA1 of hippocampal slices. Increasing numbers of theta-burst trains (1, 4, and 8 TBS) induced LTP of increasing magnitude and persistence. Inhibition of ryanodine receptors caused inhibition of weak LTP induced by 1 TBS, but had no effect on more robust forms of LTP. Inhibition of IP3 receptors inhibited moderate LTP induced by 4 TBS, but had no effect when 1 TBS or 8 TBS were used. Inhibition of L-type voltage-dependent Ca(2+) channels inhibited strong LTP induced by 8 TBS, but had no effect on weaker forms of LTP. These results show that different Ca(2+) sources have different thresholds for activation by TBS trains. Furthermore, each Ca(2+) source appears to be tuned to the induction of a different form of LTP. Such tuning could reflect an important link between different LTP induction and maintenance mechanisms. PMID- 12091551 TI - Central sensitization of nociceptive neurons in trigeminal subnucleus oralis depends on integrity of subnucleus caudalis. AB - Our recent studies have shown that application to the tooth pulp of the inflammatory irritant mustard oil (MO) produces a prolonged (>40 min) "central sensitization" reflected in neuroplastic changes in the mechanoreceptive field (RF) and response properties of nociceptive brain stem neurons in subnuclei oralis (Vo) and caudalis (Vc) of the trigeminal spinal tract nucleus. In view of the previously demonstrated ascending modulatory influence of Vc on Vo, our aim was to determine whether the Vo neuroplastic changes induced by MO application to the tooth pulp depend on an ascending influence from Vc. In chloralose/urethan anesthetized rats, MO application to the pulp produced significant increases in Vo nociceptive neuronal orofacial RF size and responses to mechanical noxious stimuli that lasted as long as 40-60 min. These changes were not affected by vehicle (saline) microinjected into Vc at 20 min after MO application, but 0.3 microl of a 5 mM CoCl(2) solution microinjected into the ipsilateral Vc produced a reversible blockade of the MO-induced Vo neuroplastic changes. A similar volume and concentration of CoCl(2) solution injected into subnucleus interpolaris of the trigeminal spinal tract nucleus did not affect the MO-induced neuroplastic changes in Vo. These findings indicate that inflammatory pulp-induced central sensitization in Vo is dependent on the functional integrity of Vc. PMID- 12091552 TI - Motor-unit synchronization alters spike-triggered average force in simulated contractions. AB - The purpose of the study was to quantify the effect of motor-unit synchronization on the spike-triggered average forces of a population of motor units. Muscle force was simulated by defining mechanical and activation characteristics of the motor units, specifying motor neuron discharge times, and imposing various levels of motor-unit synchronization. The model comprised 120 motor units. Simulations were performed for motor units 5-120 to compare the spike-triggered average responses in the presence and absence of motor-unit synchronization with the motor-unit twitch characteristics defined in the model. To synchronize motor-unit activity, selected motor-unit discharge times were adjusted; this kept the number of action potentials constant across the three levels of synchrony for each motor unit. Because there was some overlap of motor-unit twitches even at minimal discharge rates, the simulations indicated that spike-triggered averaging underestimates the twitch force of all motor units and the contraction time of motor units with contraction times longer than 49 ms. Although motor-unit synchronization increased the estimated twitch force and decreased the estimated contraction time of all motor units, spike-triggered average force changed systematically with the level of synchrony in motor units 59-120 (upper 90% of the range of twitch forces). However, the reduction in contraction time was similar for moderate and high synchrony. In conclusion, spike-triggered averaging appears to provide a biased estimate of the distribution of twitch properties for a population of motor units because twitch fusion causes an underestimation of twitch force for slow units and motor-unit synchronization causes an overestimation of force for fast motor units. PMID- 12091554 TI - Role of apamin-sensitive k(ca) channels for reticulospinal synaptic transmission to motoneuron and for the afterhyperpolarization. AB - Single motoneurons and pairs of a presynaptic reticulospinal axon and a postsynaptic motoneuron were recorded in the isolated lamprey spinal cord, to investigate the role of calcium-dependent K(+) channels (K(Ca)) during the afterhyperpolarization following the action potential (AHP), and glutamatergic synaptic transmission on the dendritic level. The AHP consists of a fast phase due to transient K(+) channels (fAHP) and a slower phase lasting 100-200 ms (sAHP), being the main determinant of spike frequency regulation. We now present evidence that the sAHP has two components. The larger part, around 80%, is abolished by superfusion of Cd(2+) (blocker of voltage-dependent Ca(2+) channels), by intracellular injection of 1,2-bis-(2-aminophenoxy)-ethane N,N,N',N'-tetraacetic acid (BAPTA; fast Ca(2+) chelator), and by apamin (selective toxin for K(Ca) channels of the SK subtype). While 80% of the sAHP is thus due to K(Ca) channels, the remaining 20% is not mediated by Ca(2+), either entering through voltage-dependent Ca(2+) channels or released from intracellular Ca(2+) stores. This Ca(2+)-independent sAHP component has a similar time course as the K(Ca) portion and is not due to a Cl(-) conductance. It may be caused by Na(+)-activated K(+) channels. Glutamatergic excitatory postsynaptic potentials (EPSPs) evoked by single reticulospinal axons give rise to a local Ca(2+) increase in the postsynaptic dendrite, mediated in part by N-methyl-D-aspartate (NMDA) receptors. The Ca(2+) levels remain elevated for several hundred milliseconds and could be expected to activate K(Ca) channels. If so, this activation should cause a local conductance increase in the dendrite that would shunt EPSPs following the first EPSP in a spike train. We have tested this in reticulospinal/motoneuronal pairs, by stimulating the presynaptic axon with spike trains at different frequencies. We compared the first EPSP and the following EPSPs in the control and after blockade with apamin. No difference was observed in EPSP amplitude or shape before and after apamin, either in normal Ringer or in Mg(2+)-free Ringer removing the voltage-dependent block of NMDA receptors. In conclusion, the local Ca(2+) entry during reticulospinal EPSPs does not cause an activation of K(Ca) channels sufficient to affect the efficacy of synaptic transmission. Thus the integration of synaptic signals at the dendritic level in motoneurons appears simpler than would otherwise have been the case. PMID- 12091553 TI - ATP-inhibition of M current in frog sympathetic neurons involves phospholipase C but not Ins P(3), Ca(2+), PKC, or Ras. AB - Suppression of the voltage-activated, noninactivating K(+) conductance (M conductance; g(M)) by muscarinic agonists, P(2Y) agonists or bradykinin increases neuronal excitability. All agonist effects are mediated, at least in part, via the Gq/(11) class of G protein. We found, using whole cell or perforated patch recording from bullfrog sympathetic B neurons that ATP-induced suppression of g(M) was attenuated by the phospholipase C (PLC) inhibitor, U73122 (IC(50) approximately 0.14 microM) but not by the inactive isomer, U73343. The ability of extracellularly applied U73122 to inhibit PLC was confirmed by its antagonism of ATP-induced elevation of intracellular Ca(2+) as measured by fura-2 photometry. ATP-induced g(M) suppression was not antagonized by the protein kinase C (PKC) inhibitor, chelerythrine (5 microM extracellular +10 microM intracellular), by the Ca(2+)-ATPase inhibitor, thapsigargin (5 microM), or by inositol trisphosphate (InsP(3)) receptor antagonists, heparin (approximaterly 300 microM) or xestospongin C (1.8 microM). The effect of ATP on g(M) was thus dependent on PLC yet independent of PKC and of InsP(3)-induced release of intracellular Ca(2+). We therefore tested the involvement of a PKC-independent action of diacylglycerol (DAG) that could occur via activation of Ras. This low-molecular weight G protein is activated following DAG binding to Ras-GRP, a neuronal Ras GTP exchange factor. However, impairment of Ras function by culturing neurons with isoprenylation inhibitors (perillic acid, 0.1 mM, or alpha-hydroxyfarnesyl phosphonic acid, 10 microM) failed to affect ATP-induced g(M) suppression. Inhibition of MEK (mitogen-activated protein kinase), a downstream target of Ras, by using PD 98059 (10 microM) was also ineffective. The transduction mechanism used by ATP to suppress g(M) in frog sympathetic neurons therefore differs from the PLC-independent mechanism used by muscarine and from the PLC and Ca(2+) dependent mechanism used by bradykinin and UTP in mammalian ganglia. The possibility remains that "lipid-signaling" mechanisms, perhaps involving PLC induced depletion of phosphatidylinositol bisphosphate, are involved in PLC mediated inhibition of g(M) by ATP in amphibian sympathetic neurons. PMID- 12091555 TI - Cortical representation of venous nociception in humans. AB - Painful sensations can be evoked by application of thermal, mechanical, and chemical stimuli to the blood vessels. The cortical substrates of these sensations are unknown. We therefore used whole-head magnetoencephalography to record cortical responses to painful laser stimuli applied cutaneously and intravenously to the dorsum of the hand in healthy human subjects. Similar to the cutaneous stimuli, venous stimulation nearly simultaneously activated the contralateral primary and the bilateral secondary somatosensory cortices. In the venous stimulation condition, all activation peaks were about 50 ms earlier than in the cutaneous stimulation condition. Locations of responses to both stimuli did not differ. These results show that the afferent volley from the veins reaches the cerebral cortex significantly earlier than that from the skin. This might be due to differences in peripheral conduction velocity. Apart from this, these findings demonstrate that venous nociception shares the cortical representation of cutaneous nociception in human somatosensory cortices. Thus the cortical representation of nociceptive processing from tissues of mesodermal and ectodermal origin appears to be similar. PMID- 12091556 TI - Interaction between visual and vestibular signals for the control of rapid eye movements. AB - To investigate interactions between voluntary and reflexive eye movements, five subjects were asked to make pro- or anti-saccades to various oblique locations cued by a head-fixed flash while being rotated sinusoidally in yaw (0.17 Hz; 73 degrees /s peak velocity) in complete darkness. Eye movements were recorded with the coil technique. In the pro-saccade task, targeting responses showed clear compensation for the intervening nystagmus, but there was a marked increase in horizontal scatter. Most quick phases directed into the hemifield opposite to the flash (away trials) were suppressed from ~100 ms onward. By contrast, quick phases directed into the hemifield of the flash (toward trials) continued virtually unabated until visually triggered saccades began to appear. From 80 ms onward, these vestibularly triggered movements showed signs of metrical modification by the visual signal. In the anti-saccade experiments, suppression of quick phases away from the flash was just as strong as in the pro-saccade experiments, and error rates in these trials were almost as low as in stationary control conditions. Suppression of quick phases directed toward the flash was a new phenomenon that emerged only in anti-saccade experiments. Since this inhibition had a late onset and was only partial, error rates in anti-saccade toward trials were very high. At short latencies, both components of most rapid eye movements were wrongly directed toward the flash. This was followed by a stage with frequent incongruent responses in which unsuppressed quick phases provoked an incorrect horizontal movement, whereas the vertical component showed a correct anti-saccade response. At still longer latencies, most responses were correct in both components. The visual modification of short-latency responses in both tasks showed that rapid eye movements could not simply be classified as either voluntary or reflexive, but suggested that signals underlying each class could merge into a compromise response. That vestibular rotation during the anti saccade task may cause a wrongly directed horizontal component resembling a quick phase, combined with a vertical component expressing a correct anti-saccade signal, reveals a remarkable independence at the component level. These observations suggest that voluntary and involuntary movements can be programmed in parallel. This behavior is explained most parsimoniously by assuming that the two signals converge at a component-coding stage of the system, rather than at a vectorial coding stage. PMID- 12091557 TI - AMRP peptides modulate a novel K(+) current in pleural sensory neurons of Aplysia. AB - Modulation of Aplysia mechanosensory neurons is thought to underlie plasticity of defensive behaviors that are mediated by these neurons. In the past, identification of modulators that act on the sensory neurons and characterization of their actions has been instrumental in providing insight into the functional role of the sensory neurons in the defensive behaviors. Motivated by this precedent and a recent report of the presence of Aplysia Mytilus inhibitory peptide-related (AMRP) neuropeptides in the neuropile and neurons of the pleural ganglia, we sought to determine whether and how pleural sensory neurons respond to the AMRPs. In cultured pleural sensory neurons under voltage clamp, AMRPs elicited a relatively rapidly developing, then partially desensitizing, outward current. The current exhibited outward rectification; in normal 10 mM K(+), it was outward at membrane potentials more positive than -80 mV but disappeared without reversing at more negative potentials. When external K(+) was elevated to 100 mM, the AMRP-elicited current reversed around -25 mV; the shift in reversal potential was as expected for a current carried primarily by K(+). In the high K(+) solution, the reversed current began to decrease at potentials more negative than -60 mV, creating a region of negative slope resistance in the I-V relationship. The AMRP-elicited K(+) current was blocked by extremely low concentrations of 4-aminopyridine (4-AP; IC(50) = 1.7 x 10(-7) M) but was not very sensitive to TEA. In cell-attached patches, AMRPs applied outside the patch thus presumably through a diffusible messenger-increased the activity of a K(+) channel that very likely underlies the macroscopic current. The single-channel current exhibited outward rectification, and the open probability of the channel decreased with hyperpolarization; together, these two factors accounted for the outward rectification of the macroscopic current. Submicromolar 4-AP included in the patch pipette blocked the channel by reducing its open probability without altering the single-channel current. Based on the characteristics of the AMRP modulated K(+) current, we conclude that it is a novel current that has not been previously described in Aplysia mechanosensory neurons. In addition to this current, two other AMRP-elicited currents, a slow, 4-AP-resistant outward current and a Na(+)-dependent inward current, were occasionally observed in the cultured sensory neurons. Responses consistent with all three currents were observed in sensory neurons in situ in intact pleural ganglia. PMID- 12091558 TI - Effect of volitional inhibition on cortical inhibitory mechanisms. AB - To investigate the effect of volitional inhibition on cortical inhibitory mechanisms, we performed transcranial magnetic stimulation (TMS) studies with a Go/NoGo reaction task in seven healthy subjects. Subjects were asked to extend their right index finger only after Go, but to remain relaxed after NoGo. Single- and paired-pulse TMS were triggered at the average reaction time for the Go response in each subject after Go or NoGo cues. Motor evoked potentials were recorded in the extensor indicis proprius (EIP) and abductor digiti minimi (ADM) muscles of right hand. Paired-pulse TMS with subthreshold conditioning stimuli at interstimulus intervals (ISIs) of 2 ms [short intracortical inhibition (SICI)] and 15 ms [intracortical facilitation (ICF)] and that with suprathreshold conditioning stimuli at ISI of 80 ms [long intracortical inhibition (LICI)] were performed in both Go/NoGo and control conditions. Inhibition of SICI was enhanced in both EIP and ADM after NoGo and was reduced only in EIP after Go. Inhibition of LICI was reduced in both muscles during both conditions, while ICF was not altered. The present results demonstrate that volitional inhibition enhances SICI but reduces LICI nonselectively. These results suggest that these two inhibitory mechanisms act differently during execution and suppression of voluntary movements. PMID- 12091559 TI - Strategies for dynamic stability during locomotion on a slippery surface: effects of prior experience and knowledge. AB - Falls due to slips are prevalent in everyday life. The purpose of this study was to determine the reactive recovery responses used to maintain dynamic stability during an unexpected slip, establish the time course of response adaptation to repeated slip perturbations, and distinguish the proactive strategies for negotiating a slippery surface. Twelve young adults participated in the study in which a slip was generated following foot contact on a set of steel free-wheeling rollers. Surface electromyographic (EMG) data were collected from rectus femoris, biceps femoris, tibialis anterior, and the medial head of gastrocnemius on the perturbed limb. Whole body kinematics were recorded using an optical imaging system: from this the center of mass, foot angle, and medial-lateral stability margins were determined. In addition, braking/loading and accelerating/unloading impulses while in contact with the rollers and the rate of loading the rollers were determined from ground reaction forces. Results demonstrate that the reactive recovery response to the first slip consisted of a rapid onset of a flexor synergy (146-199 ms), a large arm elevation strategy, and a modified swing limb trajectory. With repeated exposure to the slip perturbation, the CNS rapidly adapts within one slip trial through global changes. These changes include the attenuation of muscle response magnitude, reduced braking impulse, landing more flat-footed, and elevating the center of mass. Individuals implement a "surfing strategy" while on the rollers when knowledge of the surface condition was available before hand. Furthermore, knowledge of a slip results in a reduced braking impulse and rate of loading, a shift in medial-lateral center of mass closer to the support limb at foot contact on the rollers and a more flat foot landing. In conclusion, prior experience with the perturbations allows subsequent modification and knowledge of the surface condition results in proactive adjustments to safely traverse the slippery surface. PMID- 12091560 TI - Constraints on the source of short-term motion adaptation in macaque area MT. I. the role of input and intrinsic mechanisms. AB - Neurons in area MT, a motion-sensitive area of extrastriate cortex, respond to a step of target velocity with a transient-sustained firing pattern. The transition from a high initial firing rate to a lower sustained rate occurs over a time course of 20-80 ms and is considered a form of short-term adaptation. The present paper asks whether adaptation is due to input-specific mechanisms such as short term synaptic depression or if it results from intrinsic cellular mechanisms such as spike-rate adaptation. We assessed the contribution of input-specific mechanisms by using a condition/test paradigm to measure the spatial scale of adaptation. Conditioning and test stimuli were placed within MT receptive fields but were spatially segregated so that the two stimuli would activate different populations of inputs from the primary visual cortex (V1). Conditioning motion at one visual location caused a reduction of the transient firing to subsequent test motion at a second location. The adaptation field, estimated as the region of visual space where conditioning motion caused adaptation, was always larger than the MT receptive field. Use of the same stimulus configuration while recording from direction-selective neurons in V1 failed to demonstrate either adaptation or the transient-sustained response pattern that is the signature of short-term adaptation in MT. We conclude that the shift from transient to sustained firing in MT cells does not result from an input-specific mechanism applied to inputs from V1 because it operates over a wider range of the visual field than is covered by receptive fields of V1 neurons. We used a direct analysis of MT neuron spike trains for many repetitions of the same motion stimulus to assess the contribution to adaptation of intrinsic cellular mechanisms related to spiking. On a trial-by-trial basis, there was no correlation between number of spikes in the transient interval and the interval immediately after the transient period. This is opposite the prediction that there should be a correlation if spikes cause adaptation directly. Further, the transient was suppressed or extinguished, not delayed, in trials in which the neuron emitted zero spikes during the interval that showed a transient in average firing rate. We conclude that the transition from transient to sustained firing in neurons in area MT is caused by mechanisms that are neither input-specific nor controlled by the spiking of the adapting neuron. We propose that the short-term adaptation observed in area MT emerges from the intracortical circuit within MT. PMID- 12091561 TI - Constraints on the source of short-term motion adaptation in macaque area MT. II. tuning of neural circuit mechanisms. AB - Neurons in area MT, a motion-sensitive area of extrastriate cortex, respond to a step of target velocity with a transient-sustained firing pattern. The transition from a high initial firing rate to a lower sustained rate occurs over a time course of 20-80 ms and is considered a form of short-term adaptation. In the present paper, we compared the tuning of the adaptation to the neuron's tuning to direction and speed. The tuning of adaptation was measured with a condition/test paradigm in which a testing motion of the preferred direction and speed of the neuron under study was preceded by a conditioning motion: the direction and speed of the conditioning motion were varied systematically. The response to the test motion depended strongly on the direction of the conditioning motion. It was suppressed in almost all neurons by conditioning motion in the same direction and could be either suppressed or enhanced by conditioning motion in the opposite direction. Even in neurons that showed suppression for target motion in the nonpreferred direction, the adaptation and response direction tuning were the same. The speed tuning of adaptation was linked much less tightly to the speed tuning of the response of the neuron under study. For just more than 50% of neurons, the preferred speed of adaptation was more than 1 log unit different from the preferred response speed. Many neurons responded best when slow motions were followed by faster motions (acceleration) or vice versa (deceleration), suggesting that MT neurons may encode information about the change of target velocity over time. Finally, adaptation by conditioning motions of different directions, but not different speeds, altered the latency of the response to the test motion. The adaptation of latency recovered with shorter intervals between the conditioning and test motions than did the adaptation of response size, suggesting that latency and amplitude adaptation are mediated by separate mechanisms. Taken together with the companion paper, our data suggest that short term motion adaptation in MT is a consequence of the neural circuit in MT and is not mediated by either input-specific mechanisms or intrinsic mechanisms related to the spiking of individual neurons. The circuit responsible for adaptation is tuned for both speed and direction and has the same direction tuning as the circuit responsible for the initial response of MT neurons. PMID- 12091562 TI - Optical approaches to functional organization of glossopharyngeal and vagal motor nuclei in the embryonic chick hindbrain. AB - We investigated the functional organization of the glossopharyngeal and vagal motor nuclei during embryogenesis using multiple-site optical recording with a fast voltage-sensitive dye. Intact brain stem preparations with glossopharyngeal and vagus nerves were dissected from 4- to 8-day-old chick embryos. Electrical responses evoked by glossopharyngeal/vagus nerve stimulation were optically recorded from many loci of the stained preparations. In 4- to 6-day-old preparations, action potential-related fast spikelike signals were detected from the nucleus of the glossopharyngeal nerve and the dorsal motor nucleus of the vagus nerve. Contour line maps of the signal amplitude showed multiple-peak patterns, suggesting that the neurons and/or their activity were not uniformly distributed within the nuclei at early developmental stages. As development proceeded from 4 to 6 days, the peaks fused with each other and the number of peaks decreased gradually. In most 7- and 8-day-old preparations, only a single peak was identified in the nuclei, and the distribution of the signal amplitude formed a layered pattern surrounding the peak-signal area. These results suggest that functional organization of the motor nuclei in the embryonic hindbrain changes dynamically with development, resulting in a rearrangement of functional nuclear cores from multiple-peaks to a single peak. PMID- 12091564 TI - NRSF causes cAMP-sensitive suppression of sodium current in cultured hippocampal neurons. AB - The neuron restrictive silencer factor (NRSF/REST) has been shown to bind to the promoters of many neuron-specific genes and is able to suppress transcription of Na(+) channels in PC12 cells, although its functional effect in terminally differentiated neurons is unknown. We constructed lentiviral vectors to express NRSF as a bicistronic message with green fluorescent protein (GFP) and followed infected hippocampal neurons in culture over a period of 1-2 wk. NRSF-expressing neurons showed a time-dependent suppression of Na(+) channel function as measured by whole cell electrophysiology. Suppression was reversed or prevented by the addition of membrane-permeable cAMP analogues and enhanced by cAMP antagonists but not affected by increasing protein expression with a viral enhancer. Secondary effects, including altered sensitivity to glutamate and GABA and reduced outward K(+) currents, were duplicated by culturing GFP-infected control neurons in TTX. The striking similarity of the phenotypes makes NRSF potentially useful as a genetic "silencer" and also suggests avenues of further exploration that may elucidate the transcription factor's in vivo role in neuronal plasticity. PMID- 12091563 TI - Differential subcellular localization of the two alternatively spliced isoforms of the Kv3.1 potassium channel subunit in brain. AB - Voltage-gated K(+) channels containing pore-forming subunits of the Kv3 subfamily have specific roles in the fast repolarization of action potentials and enable neurons to fire repetitively at high frequencies. Each of the four known Kv3 genes encode multiple products by alternative splicing of 3' ends resulting in the expression of K(+) channel subunits differing only in their C-terminal sequence. The alternative splicing does not affect the electrophysiological properties of the channels, and its physiological role is unknown. It has been proposed that one of the functions of the alternative splicing of Kv3 genes is to produce subunit isoforms with differential subcellular membrane localizations in neurons and differential modulation by signaling pathways. We investigated the role of the alternative splicing of Kv3 subunits in subcellular localization by examining the brain distribution of the two alternatively spliced versions of the Kv3.1 gene (Kv3.1a and Kv3.1b) with antibodies specific for the alternative spliced C-termini. Kv3.1b proteins were prominently expressed in the somatic and proximal dendritic membrane of specific neuronal populations in the mouse brain. The axons of most of these neurons also expressed Kv3.1b protein. In contrast, Kv3.1a proteins were prominently expressed in the axons of some of the same neuronal populations, but there was little to no Kv3.1a protein expression in somatodendritic membrane. Exceptions to this pattern were seen in two neuronal populations with unusual targeting of axonal proteins, mitral cells of the olfactory bulb, and mesencephalic trigeminal neurons, which expressed Kv3.1a protein in dendritic and somatic membrane, respectively. The results support the hypothesis that the alternative spliced C-termini of Kv3 subunits regulate their subcellular targeting in neurons. PMID- 12091565 TI - Temporally staggered forelimb stimulation modulates barrel cortex optical intrinsic signal responses to whisker stimulation. AB - Characterization of neurovascular relationships is critical to accurate interpretation of functional neuroimaging data. We have previously observed spatial uncoupling of optical intrinsic signal imaging (OIS) and evoked potential (EP) responses in rodent barrel cortex following simultaneous whisker and forelimb stimulation, leading to changes in OIS response magnitude. To further test the hypothesis that this uncoupling may have resulted from "passive" overspill of perfusion-related responses between functional regions, we conducted the present study using temporally staggered rather than simultaneous whisker and forelimb stimulation. This paradigm minimized overlap of neural responses in barrel cortex and forelimb primary somatosensory cortex (SI), while maintaining overlap of vascular response time courses between regions. When contrasted with responses to 1.5-s lone-whisker stimulation, staggered whisker and forelimb stimulation resulted in broadening of barrel cortex OIS response time course in the temporal direction of forelimb stimulation. OIS response peaks were also temporally shifted toward the forelimb stimulation period; time-to-peak was shorter (relative to whisker stimulus onset) when forelimb stimulation preceded whisker stimulation and longer when forelimb stimulation followed whisker stimulation. In contrast with OIS and EP magnitude decreases previously observed during simultaneous whisker/forelimb stimulation, barrel cortex OIS response magnitude increased during staggered stimulation and no detectable changes in underlying EP activity were observed. Spatial extent of barrel cortex OIS responses also increased during staggered stimulation. These findings provide further evidence for spatial uncoupling of OIS and EP responses, and emphasize the importance of temporal stimulus properties on the effects of this uncoupling. It is hypothesized that spatial uncoupling is a result of passive overspill of perfusion-related responses into regions distinct from those which are functionally active. It will be important to consider potential influences of this uncoupling when designing and interpreting functional imaging studies that use hemodynamic responses to infer underlying neural activity. PMID- 12091566 TI - Auditory-visual interactions subserving goal-directed saccades in a complex scene. AB - This study addresses the integration of auditory and visual stimuli subserving the generation of saccades in a complex scene. Previous studies have shown that saccadic reaction times (SRTs) to combined auditory-visual stimuli are reduced when compared with SRTs to either stimulus alone. However, these results have been typically obtained with high-intensity stimuli distributed over a limited number of positions in the horizontal plane. It is less clear how auditory-visual interactions influence saccades under more complex but arguably more natural conditions, when low-intensity stimuli are embedded in complex backgrounds and distributed throughout two-dimensional (2-D) space. To study this problem, human subjects made saccades to visual-only (V-saccades), auditory-only (A-saccades), or spatially coincident auditory-visual (AV-saccades) targets. In each trial, the low-intensity target was embedded within a complex auditory-visual background, and subjects were allowed over 3 s to search for and foveate the target at 1 of 24 possible locations within the 2-D oculomotor range. We varied systematically the onset times of the targets and the intensity of the auditory target relative to background [i.e., the signal-to-noise (S/N) ratio] to examine their effects on both SRT and saccadic accuracy. Subjects were often able to localize the target within one or two saccades, but in about 15% of the trials they generated scanning patterns that consisted of many saccades. The present study reports only the SRT and accuracy of the first saccade in each trial. In all subjects, A saccades had shorter SRTs than V-saccades, but were more inaccurate than V saccades when generated to auditory targets presented at low S/N ratios. AV saccades were at least as accurate as V-saccades but were generated at SRTs typical of A-saccades. The properties of AV-saccades depended systematically on both stimulus timing and S/N ratio of the auditory target. Compared with unimodal A- and V-saccades, the improvements in SRT and accuracy of AV-saccades were greatest when the visual target was synchronous with or leading the auditory target, and when the S/N ratio of the auditory target was lowest. Further, the improvements in saccade accuracy were greater in elevation than in azimuth. A control experiment demonstrated that a portion of the improvements in SRT could be attributable to a warning-cue mechanism, but that the improvements in saccade accuracy depended on the spatial register of the stimuli. These results agree well with earlier electrophysiological results obtained from the midbrain superior colliculus (SC) of anesthetized preparations, and we argue that they demonstrate multisensory integration of auditory and visual signals in a complex, quasi-natural environment. A conceptual model incorporating the SC is presented to explain the observed data. PMID- 12091567 TI - Spatial structure and symmetry of simple-cell receptive fields in macaque primary visual cortex. AB - I present measurements of the spatial structure of simple-cell receptive fields in macaque primary visual cortex (area V1). Similar to previous findings in cat area 17, the spatial profile of simple-cell receptive fields in the macaque is well described by two-dimensional Gabor functions. A population analysis reveals that the distribution of spatial profiles in primary visual cortex lies approximately on a one-parameter family of filter shapes. Surprisingly, the receptive fields cluster into even- and odd-symmetry classes with a tendency for neurons that are well tuned in orientation and spatial frequency to have odd symmetric receptive fields. The filter shapes predicted by two recent theories of simple-cell receptive field function, independent component analysis and sparse coding, are compared with the data. Both theories predict receptive fields with a larger number of subfields than observed in the experimental data. In addition, these theories do not generate receptive fields that are broadly tuned in orientation and low-pass in spatial frequency, which are commonly seen in monkey V1. The implications of these results for our understanding of image coding and representation in primary visual cortex are discussed. PMID- 12091568 TI - Differentiating noxious- and innocuous-related activation of human somatosensory cortices using temporal analysis of fMRI. AB - The role of the somatosensory cortices (SI and SII) in pain perception has long been in dispute. Human imaging studies demonstrate activation of SI and SII associated with painful stimuli, but results have been variable, and the functional relevance of any such activation is uncertain. The present study addresses this issue by testing whether the time course of somatosensory activation, evoked by painful heat and nonpainful tactile stimuli, is sufficient to discriminate temporal differences that characterize the perception of these stimulus modalities. Four normal subjects each participated in three functional magnetic resonance imaging (fMRI) sessions, in which painful (noxious heat 45-46 degrees C) and nonpainful test stimuli (brushing at 2 Hz) were applied repeatedly (9-s stimulus duration) to the left leg in separate experiments. Activation maps were generated comparing painful to neutral heat (35 degrees C) and nonpainful brushing to rest. Directed searches were performed in SI and SII for sites reliably activated by noxious heat and brush stimuli, and stimulus-dependent regions of interest (ROI) were then constructed for each subject. The time course, per stimulus cycle, was extracted from these ROIs and compared across subjects, stimulus modalities, and cortical regions. Both innocuous brushing and noxious heat produced significant activation within contralateral SI and SII. The time course of brush-evoked responses revealed a consistent single peak of activity, approximately 10 s after the onset of the stimulus, which rapidly diminished upon stimulus withdrawal. In contrast, the response to heat pain in both SI and SII was characterized by a double-peaked time course in which the maximum response (the 2nd peak) was consistently observed approximately 17 s after the onset of the stimulus (8 s following termination of the stimulus). This prolonged period of activation paralleled the perception of increasing pain intensity that persists even after stimulus offset. On the other hand, the temporal profile of the initial minor peak in pain-related activation closely matched that of the brush-evoked activity, suggesting a possible relationship to tactile components of the thermal stimulation procedure. These data indicate that both SI and SII cortices are involved in the processing of nociceptive information and are consistent with a role for these structures in the perception of temporal aspects of pain intensity. PMID- 12091570 TI - Nigrostriatal lesion and dopamine agonists affect firing patterns of rodent entopeduncular nucleus neurons. AB - Altered activity of the entopeduncular nucleus, the rodent homologue of the globus pallidus internal segment in primates, is thought to mediate behavioral consequences of midbrain dopamine depletion in rodents. Few studies, however, have examined dopaminergic modulation of spiking activity in this nucleus. This study characterizes changes in entopeduncular neuronal activity after nigrostriatal dopaminergic lesion and the effects of systemic treatment with selective D(1) (SKF 38393) and D(2) (quinpirole) agonists in lesioned rats. Extracellular single-unit recordings were performed in awake immobilized rats, either in neurologically intact animals (n = 42) or in animals that had received unilateral 6-hydroxydopamine infusion into the medial forebrain bundle several weeks previously (n = 35). Nigrostriatal lesion altered baseline activity of entopeduncular neurons in several ways. Interspike interval distributions had significantly decreased modes and significantly increased coefficient of variation, skewness and kurtosis; yet interspike interval mean (the inverse of firing rate) was not affected. Also, spectral analysis of autocorrelograms indicated that lesion significantly reduced the incidence of regular-spiking neurons and increased the incidence of neurons with 4-18 Hz oscillations. Dopamine agonist treatment reversed some lesion-induced effects: quinpirole reversed changes in interspike interval distribution mode and coefficient of variation, while combined quinpirole and SKF 38393 blocked the appearance of 4-18 Hz oscillations. However, no agonist treatment normalized all aspects of entopeduncular activity. Additionally, inhibition of firing rates by D(1) or combined D(1)/D(2) receptor activation indicated that dopamine agonists affected the overall level of entopeduncular activity in a manner similar to that found in the substantia nigra pars reticulata and globus pallidus internal segment after dopamine neuron lesion. These data demonstrate that lesion of the nigrostriatal tract leads to modifications of several aspects of firing pattern in the rodent entopeduncular nucleus and so expand on similar findings in the rodent substantia nigra pars reticulata and in the globus pallidus internal segment in humans and nonhuman primates. The results support the view that dysfunction in the basal ganglia after midbrain dopamine neuron loss relates more consistently to abnormal activity patterns than to net changes in firing rate in the basal ganglia output nuclei, while overall decreases in firing rate in these structures may play a more important role in adverse motor reactions to dopamine agonist treatments. PMID- 12091569 TI - Chronic interleukin-6 exposure alters electrophysiological properties and calcium signaling in developing cerebellar purkinje neurons in culture. AB - The cytokine interleukin-6 (IL-6) is chronically expressed at elevated levels within the CNS in many neurological disorders and may contribute to the histopathological, pathophysiological, and cognitive deficits associated with such disorders. However, the effects of chronic IL-6 exposure on neuronal function in the CNS are largely unknown. Therefore using intracellular recording and calcium imaging techniques, we investigated the effects of chronic IL-6 exposure on the physiological properties of cerebellar Purkinje neurons in primary culture. Two weeks of exposure to 1,000 units/ml (U/ml) IL-6 resulted in altered electrophysiological properties of Purkinje neurons, including a significant reduction in action potential generation, an increase in input resistance, and an enhanced electrical response to the ionotropic glutamate receptor agonist, alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) compared with untreated neurons. Lower concentrations of IL-6 (100 and 500 U/ml) had no effects on these electrophysiological parameters. However, neurons exposed to 500 U/ml chronic IL-6 resulted in significantly elevated resting levels of intracellular calcium as well as an increase in the intracellular calcium signal of Purkinje neurons in response to AMPA, effects not observed in neurons exposed to 1,000 U/ml chronic IL-6. Morphometric analysis revealed a lack of gross structural changes following chronic IL-6 treatment, such as in the number, size, and extent of dendritic arborization of Purkinje neurons in culture. Using immunohistochemistry, we found that cultured Purkinje neurons express both the IL 6 receptor and its intracellular signaling subunit, gp130, indicating that IL-6 may act directly on Purkinje neurons to alter their physiological properties. The present data show that chronic exposure to elevated levels of IL-6, such as occurs in various neurological diseases, produces alterations in several important physiological properties of Purkinje neurons and that these changes occur in the absence of neuronal toxicity, damage, or death. The results support the hypothesis that chronic IL-6 exposure can disrupt normal CNS function and thereby contribute to the pathophysiology associated with many neurological diseases. PMID- 12091571 TI - Nitric oxide as an anterograde neurotransmitter in the trigeminal motor pool. AB - We demonstrate the presence of nitric oxide synthase containing fibers within the guinea pig trigeminal motor nucleus and describe the effects of nitric oxide (NO) on trigeminal motoneurons. Using immunohistochemical techniques, we observed nitrergic fibers displaying varicosities and giving rise to bouton-like structures in apposition to retrogradely labeled motoneuron processes, most of which were dendrites. NO-donors evoked a membrane depolarization (mean 7.5 mV) and a decrease in rheobase (mean 38%). These substances also evoked an apparent increase in an hyperpolarization-activated cationic current (I(H)). These changes were not accompanied by any modification of the motoneurons' input resistance or time constant. The effects were suppressed by blocking the cytosolic guanlyate cyclase. A membrane-permeant cyclic guanosine 3,5'-monophosphate (cGMP) analogue mimicked the effects of NO. There was a considerable increase in synaptic activity following NO-donors or db-cGMP application. Tetrodotoxin supressed the increase in synaptic activity evoked by NO-donors. The histological and electrophysiological evidence, taken together, indicates the existence of a nitrergic system able to modulate trigeminal motoneurons under yet unknown physiological conditions. PMID- 12091573 TI - Neural activity in human primary motor cortex areas 4a and 4p is modulated differentially by attention to action. AB - The mechanisms underlying attention to action are poorly understood. Although distracted by something else, we often maintain the accuracy of a movement, which suggests that differential neural mechanisms for the control of attended and nonattended action exist. Using functional magnetic resonance imaging (fMRI) in normal volunteers and probabilistic cytoarchitectonic maps, we observed that neural activity in subarea 4p (posterior) within the primary motor cortex was modulated by attention to action, while neural activity in subarea 4a (anterior) was not. The data provide the direct evidence for differential neural mechanisms during attended and unattended action in human primary motor cortex. PMID- 12091572 TI - A model of spike-timing dependent plasticity: one or two coincidence detectors? AB - In spike-timing dependent plasticity (STDP), synapses exhibit LTD or LTP depending on the order of activity in the presynaptic and postsynaptic cells. LTP occurs when a single presynaptic spike precedes a postsynaptic one (a positive interspike interval, or ISI), while the reverse order of activity (a negative ISI) produces LTD. A fundamental question is whether the "standard model" of plasticity in which moderate increases in Ca(2+) influx through the N-methyl-D aspartate (NMDA) channels induce LTD and large increases induce LTP, can account for the order and interval sensitivity of STDP. To examine this issue we developed a model that captures postsynaptic Ca(2+) influx dynamics and the associativity of the NMDA receptors. While this model can generate both LTD and LTP, it predicts that LTD will be observed at both negative and positive ISIs. This is because longer and longer positive ISIs induce monotonically decreasing levels of Ca(2+), which eventually fall into the same range that produced LTD at negative ISIs. A second model that incorporated a second coincidence detector in addition to the NMDA receptor generated LTP at positive intervals and LTD only at negative ones. Our findings suggest that a single coincidence detector model based on the standard model of plasticity cannot account for order-specific STDP, and we predict that STDP requires two coincidence detectors. PMID- 12091574 TI - Presynaptic suppression of dorsal horn inhibitory transmission by mu-opioid receptors. AB - Opioids modify sensory experience at many levels in the CNS. The mechanisms of this action, including the ways opioid receptors affect synaptic transmission, are not yet fully understood. Here we show that the selective activation of mu opioid receptors suppressed inhibitory transmission between spinal cord dorsal horn neurons in vitro. mu-Opioid receptor activation reduced evoked inhibitory postsynaptic current (eIPSC) amplitude by acting presynaptically, because it altered the paired-pulse ratio, did not affect GABA-evoked currents, and decreased miniature IPSC (mIPSC) frequency. The mechanism of this effect was independent both of presynaptic Ca(2+) entry and of the pathway linking presynaptic kainate (KA) receptors to suppression of inhibitory transmission in the same cells. These data identify mu-opioid receptors as important presynaptic modulators of dorsal horn inhibitory transmission. PMID- 12091575 TI - Paradoxical anti-epileptic effects of a GluR5 agonist of kainate receptors. AB - Kainate generates in adult hippocampal neurons a seizure but also a massive excitation of interneurons and a dramatic increase of the inhibitory drive that impinges on principal cells. This "overinhibition" is largely mediated by GluR5 containing kainate receptors that are enriched on interneurons. Here, using the neonatal intact hippocampus in vitro and the triple chamber, we first show that this mechanism is fully operative in neonatal neurons. We then report that application to one hippocampus of (RS)-2-amino-3-(5-tert-butyl-3-hydroxy-4 isoxazolyl)propionic acid-a relatively selective agonist of GluR5 containing kainate receptors-depresses the propagation of seizure generated in the opposite hippocampus by a convulsive agent. We conclude that the selective excitation of interneurons by GluR5-containing kainate receptor agonists opens a new therapeutic approach for the epilepsies. PMID- 12091576 TI - Short- and long-term plasticity of the perforant path synapse in hippocampal area CA3. AB - The direct perforant path (PP) projection to CA3 is a major source of cortical input to the hippocampal region, yet relatively little is known about the basic properties of physiology and plasticity in this pathway. We tested whether PP long-term potentiation (LTP) in CA3 possesses the Hebbian property of associativity; i.e., whether the firing of fibers of different orders can induce PP LTP. We stimulated PP with weak trains of high-frequency stimulation (HFS), which by itself was below the threshold for LTP induction. The identical HFS was effective in inducing LTP when the mossy fiber pathway (MF) was activated simultaneously, thus demonstrating associative plasticity between the two pathways. We also demonstrated associative LTP between PP and recurrent collateral fibers (RC). PP LTP was blocked by the N-methyl-D-aspartate receptor (NMDAR) antagonist 2-amino-5-phosphonovaleric acid in both the associative and homosynaptic induction conditions. Neither MF nor RC fiber HFS alone resulted in permanent changes in PP field excitatory postsynaptic potential (fEPSP) amplitude. However, HFS delivered to either MF or RC alone led to transient heterosynaptic depression of the PP fEPSP. Our results support the conceptual framework that regards CA3 as an autoassociative memory network in which efficient retrieval of previously stored activity patterns is mediated by associative plasticity of the PP synapse. PMID- 12091577 TI - Cannabinoids depress inhibitory synaptic inputs received by layer 2/3 pyramidal neurons of the neocortex. AB - Using whole cell voltage-clamp recordings we investigated the effects of a synthetic cannabinoid (WIN55,212-2) on inhibitory inputs received by layer 2/3 pyramidal neurons in slices of the mouse auditory cortex. Activation of the type 1 cannabinoid receptor (CB1R) with WIN55,212-2 reliably reduced the amplitude of GABAergic inhibitory postsynaptic currents evoked by extracellular stimulation within layer 2/3. The suppression of this inhibition was blocked and reversed by the highly selective CB1R antagonist AM251, confirming a CB1R-mediated inhibition. Pairing evoked inhibitory postsynaptic currents (IPSCs) at short interstimulus intervals while applying WIN55,212-2 resulted in an increase in paired-pulse facilitation suggesting that the probability of GABA release was reduced. A presynaptic site of cannabinoid action was verified by an observed decrease in the frequency with no change in the amplitude or kinetics of action potential-independent postsynaptic currents (mIPSCs). When Cd(2+) was added or Ca(2+) was omitted from the recording solution, the remaining fraction of Ca(2+) independent mIPSCs did not respond to WIN55,212-2. These data suggest that cannabinoids are capable of suppressing the inhibition of neocortical pyramidal neurons by depressing Ca(2+)-dependent GABA release from local interneurons. PMID- 12091578 TI - Auditory-somatosensory multisensory processing in auditory association cortex: an fMRI study. AB - Using high-field (3 Tesla) functional magnetic resonance imaging (fMRI), we demonstrate that auditory and somatosensory inputs converge in a subregion of human auditory cortex along the superior temporal gyrus. Further, simultaneous stimulation in both sensory modalities resulted in activity exceeding that predicted by summing the responses to the unisensory inputs, thereby showing multisensory integration in this convergence region. Recently, intracranial recordings in macaque monkeys have shown similar auditory-somatosensory convergence in a subregion of auditory cortex directly caudomedial to primary auditory cortex (area CM). The multisensory region identified in the present investigation may be the human homologue of CM. Our finding of auditory somatosensory convergence in early auditory cortices contributes to mounting evidence for multisensory integration early in the cortical processing hierarchy, in brain regions that were previously assumed to be unisensory. PMID- 12091579 TI - Bradykinin lowers the threshold temperature for heat activation of vanilloid receptor 1. AB - Bradykinin (BK) is an inflammatory mediator that plays a pivotal role in pain and hyperalgesia to heat in inflamed tissues by exciting nociceptors and sensitizing them to heat through activation of protein kinase C (PKC). It has been suggested that the capsaicin receptor (VR1), a nociceptor-specific cation channel sensitive to noxious heat, protons, and capsaicin, is a channel that is modified by BK in these effects. In this study, we examined how BK modulates the activity of VR1. We measured VR1 currents using the patch-clamp technique in human embryonic kidney-derived (HEK293) cells expressing VR1 and B2 BK receptor. We found that BK lowered the threshold temperature for activation of VR1 currents in HEK cells down to well below the physiological body temperature in a concentration dependent manner through PKC activation. We also demonstrated that in capsaicin sensitive dorsal root ganglion (DRG) neurons the activation threshold of heat induced current, which is considered to be VR-1 mediated, was lowered by BK and that this effect was also mediated by PKC. These data further support the supposition that modulation of VR1 is a mechanism for the BK-induced excitation of nociceptors and their sensitization to heat. PMID- 12091582 TI - Test-retest reliability of the Work Ability Index questionnaire. AB - The goal of the study was to assess the test-retest reliability of the Work Ability Index (WAI) questionnaire. Reliability was tested using a test-retest design with a 4 week interval between measurements. Valid data were collected among 97 elderly construction workers aged 40 years and older. We analysed the test-retest reliability of the WAI score itself (range 7-49 points) and classification in one of the four WAI categories based on this score: poor (7-27 points); moderate (28-36 points); good (37-43 points); and excellent work ability (44-49 points). Exactly the same WAI score on both measurements was reported by 25% of the subjects and 95% of the individual differences between measurements were found to be <6.86 points (two times standard deviation). Despite the individual changes between measurements, no significant difference was reported in the mean WAI score at group level between test and retest measurements (40.4 versus 39.9). The percentage of observed agreement for the classification of subjects in one of the four WAI categories on both measurements equalled 66%. The results of this study provided evidence of an acceptable test-retest reliability of the classification of subject's work ability by means of the WAI questionnaire. At group level, the mean WAI score and classification into WAI categories were found to be a stable measure over a 4 week interval. These results give additional support for the applicability of the questionnaire in occupational health research and the daily practice of occupational health care. PMID- 12091583 TI - Impairment of colour vision in patients with n-hexane exposure-dependent toxic polyneuropathy. AB - The aim of this study was to investigate the effects of n-hexane on visual function and to determine the duration of any symptoms related to workplace exposure. The study involved 26 workers diagnosed as having polyneuropathy following n-hexane exposure. The FM-100 Hue test was used to determine colour discrimination in study volunteers. Their results were compared with a control group of 50 people who had not been exposed to n-hexane. The mean total error score for the exposed group was 168.3 (SD = 70.5) for the right eye and 181.5 (SD = 103.0) for the left eye. The mean total error scores for the control group for the right and left eyes were 36.0 (SD = 19.8) and 35.6 (SD = 18.2), respectively. Differences between total and partial error scores for exposed and control groups were statistically significant (P < 0.001). These results may indicate a relationship between n-hexane exposure and development of defects in colour vision, and would support a recommendation for periodic assessment of workers exposed to n-hexane and chemically related solvents. PMID- 12091584 TI - Chronic fatigue among bank workers in Brazil. AB - The present study examines the prevalence of chronic fatigue (CF) among bank workers in Brazil and possible associations with gender and working conditions. The study sample included all 735 workers from the department of data processing of a state bank. CF was assessed using the Chalder Fatigue Scale. Working conditions and socio-demographic, socio-economic and psychosocial factors at work were analysed. Psychiatric symptoms were measured with the SRQ-20. The overall estimate of the prevalence of CF was 8.7% [95% confidence intervals (95% CI) = 6.4-10.9%]: 7.8% (95% CI = 5.5-10.7%) among men and 11.0% (95% CI = 6.7-16.9%) among women. The male-female difference was not statistically significant, even after adjusting for minor psychiatric disorders. The overall prevalence of CF without minor psychiatric disorders was 4.5% (95% CI = 2.7-6.3%): 3.9% (95% CI = 1.9-5.9%) among men and 6.4% (95% CI = 2.0-10.1%) among women. In the final model, risk factors for CF were fast work speed [odds ratio (OR) = 3.5], dissatisfaction at work (OR = 3.1), minor psychiatric disorders (OR = 6.8), and medium (OR = 1.8) and heavy domestic workload (OR = 12.0). CF is common among these bank workers and is associated with psychosocial factors at work. Particularly among women, domestic workload, marital status and the presence of young children were associated with CF in the stratified analysis. Domestic workload may add physical and mental stress, putting employees at risk for CF from overload, or CF may cause workers to perceive domestic work as heavy. PMID- 12091585 TI - An analysis of the causes of mortality among seafarers in the British merchant fleet (1986-1995) and recommendations for their reduction. AB - This study was designed to establish the manner and causes of all fatalities among seafarers who were serving in British merchant shipping between 1 January 1986 and 31 December 1995. Other main aims were to compare mortality rates with workers in other industries and with seafarers in other merchant fleets, and to discuss recommendations for the prevention of occupational mortality. This study was based upon official mortality files with a population of 253 919 seafarer years at risk. There was a total of 252 fatalities in British merchant shipping during the study period. One hundred and seven of the 252 deaths were due to natural causes, of which 93 (87%) were caused by cardiovascular disease. The relative risk of mortality due to accidents at work (96 fatalities) was 23.9 times higher than for all workers in Great Britain during the same time period (95% confidence interval = 14.0-40.7). The risk of a fatal accident in merchant shipping was also much higher than in other British industrial sectors, such as agriculture, forestry and fishing (relative risk = 4.5), construction (5.5), and manufacturing (22.0). Seafaring was identified as a dangerous occupation compared with shore-based employment. Many of the fatal accidents at work were caused by hazardous working practices and were often preventable, as were the many off-duty accidents and drowning that were caused by seafarers falling into docks when encountering hazardous access to their ships from the shore. Seafarers are also at special risk from acute illnesses since they lack direct access to specialist medical care. PMID- 12091586 TI - Respiratory health effects of long-term exposure to different chromium species in stainless steel production. AB - The aim of this study was to determine whether occupational exposure to chromite, trivalent chromium (Cr(3+)) or hexavalent chromium (Cr(6+)) causes respiratory diseases, an excess of respiratory symptoms, a decrease in pulmonary function or signs of pneumoconiosis among workers in stainless steel production. Altogether, 203 exposed workers and 81 referents with an average employment of 23 years were investigated for indicators of respiratory health on two occasions, in 1993 and in 1998. Data collection with a self-administered questionnaire, flow volume spirometry, measurement of diffusing capacity, chest radiography and laboratory tests were carried out by a mobile research unit. Exposure to different chromium species and other metals was monitored regularly and studied separately. No adverse respiratory health effects were observed in the group exposed to Cr(6+), either in comparison with the control group in the first cross-sectional study or during the additional 5 year follow-up. Among the Cr (3+) exposed people, the production of phlegm, shortness of breath and breathlessness on exertion were significantly more frequent than in the control group, but the frequency of the symptoms did not increase during the follow-up; no differences were observed in the lung function tests and the radiographic findings did not progress. In the chromite group, the prevalence of breathlessness on exertion was higher than in the control group. However, in the follow-up, the occurrence of symptoms did not differ from 1993 to 1998. In the first study, most parameters of lung function were lower among the smokers in the chromite group than among the smoking controls, but in 1998 the difference was less marked. An average exposure time of 23 years in modern ferrochromium and stainless steel production and low exposure to dusts and fumes containing Cr(6+), Cr(3+), nickel and molybdenum do not lead to respiratory changes detectable by lung function tests or radiography. The workers exposed to Cr(3+) had more respiratory symptoms than those in the control group. The workers in the chromite mine had lower lung function test results than the control group due to earlier exposure to higher dust concentrations. PMID- 12091587 TI - White blood cell count and clustered features of metabolic syndrome in Japanese male office workers. AB - We assessed the association of white blood cell (WBC) count with different components of metabolic syndrome (MS)-obesity, hypertension, hypercholesterolemia, low high-density lipoprotein cholesterol levels, hypertriglyceridemia, high fasting plasma glucose levels and hyperuricemia-in 5275 Japanese male office workers aged 23-59 years. After controlling for age, smoking and alcohol intake, the relative risks for the presence of 1, 2, 3, 4 and > or =5 features of MS compared with the lowest quintile of WBC count increased in a dose-dependent manner as WBC count increased (P for trend < 0.001 for all) and the increased relative risks for clustered features of MS were more pronounced as the number of features of MS increased. The WBC count increments in subjects with 1, 2, 3, 4 and > or =5 features of MS were 0.28, 0.45, 0.68, 0.76 and 1.40 x10(9) cells/l, respectively, compared with the subjects without features of MS (P for trend < 0.001). These findings indicate a strong association between WBC count and clustered features of MS in middle-aged Japanese men. PMID- 12091589 TI - Photoretinitis: an underestimated occupational injury? AB - Non-ionizing radiation, which is produced in large amounts by welding arcs, may induce photophthalmia, keratoconjunctivitis and cataracts. Retinal injuries resulting from exposure to electric welding arcs have been reported, but such injuries are not commonly seen and may be misdiagnosed. A case is described of bilateral maculopathy in a millwright exposed to metal arc inert gas-shielded welding and oxygen lance light. Insurance adjudicators denied his claim, as they did not acknowledge the cause-and-effect relationship between welding and retinopathy. Welding emits a wide spectrum of radiation, ranging from IR to UV and beyond. UV and far-IR radiation are adsorbed by the cornea and the lens, whereas visible light and near-IR radiation penetrate to the retina. According to the intensity and time of exposure, they may cause thermal or photochemical retinal damage, which may be permanent and sight-threatening. Workers covered by compulsory collective insurance should be eligible for compensation in every case of welding light-induced retinal damage. PMID- 12091588 TI - Superior orbital 'petrositis' and late ischaemic monocular blindness induced by intense UV radiation exposure. AB - Many physical agents cause neuropathies. The most common are chronic pressure, vibration and temperature. In general, these lesions occur at work, as a result of accidents or through chronic exposure to the physical agent. Radiation leading to peripheral neuropathy is also related to radiotherapy in cancer treatment, as an undesirable side-effect. We present here a case report of short, intense UV radiation exposure at work, leading to delayed-onset ocular neuropathy. A clear cause-effect relationship is shown, demonstrated using magnetic resonance imaging scans. We suggest that the mechanism was thermal and ischaemic. PMID- 12091590 TI - Preventable exposure to trimethyl tin chloride: a case report. AB - Trimethyl tin chloride (TMTC) is a highly toxic organotin compound that affects four main target organs: the brain, liver, immune system and skin. Exposure can occur by inhalation, ingestion or direct skin absorption. Trimethyl tin is but one of many hazardous substances with potentially serious health consequences to which individuals working in research laboratories may be exposed. We report a preventable case of TMTC exposure. Better understanding of the Canadian Workplace Hazardous Materials Information System (WHMIS) legislation and its applicability to the research laboratory situation would prevent such unnecessary exposure to hazardous substances. PMID- 12091594 TI - A rationale for an individualized haemoglobin target. AB - Despite the use of recombinant human erythropoietin (rh-EPO, epoetin) for more than a decade in treating renal anaemia, there is still considerable debate over optimal target haemoglobin (Hb) levels. Current European and North American guidelines that are based on decade-old trials aim for partial anaemia correction, with a subnormal target Hb concentration. More recent randomized clinical trials examining the effect of normalizing Hb levels have produced conflicting results. A study in the USA, in patients with existing congestive heart failure or ischaemic heart disease, showed an unexpected rise in cardiac mortality and haemodialysis access failure with higher Hb levels. In contrast, three other studies (in Australia, Spain and Canada) that normalized Hb levels in healthier dialysis patients observed improvements in quality of life and exercise capacity and a slower progression of left ventricular dilatation, without an unacceptable increase in the incidence of adverse effects. These studies indicate that, while higher Hb levels may be detrimental to patients with pre-existing cardiac disease, healthier patients benefit from normalized Hb levels. Thus, there is no clear scientific rationale for setting a single Hb target for all patients, and individualized treatment targets would appear to be a more logical and patient-centred approach. PMID- 12091595 TI - A rationale for an individualized administration frequency of epoetin beta: a clinical perspective. AB - The efficacy and safety of intravenous (i.v.) and subcutaneous recombinant human erythropoietin (rh-EPO, epoetin) for the treatment of renal anaemia is well established. Subcutaneous administration of epoetin is recommended by current guidelines because it is more flexible and convenient than i.v. dosing, enables dose reduction of epoetin and results in cost benefits. Subcutaneous administration of epoetin beta permits flexible dosing regimens that can be tailored to patients' individual needs. This was demonstrated in a recent, large, randomized, controlled trial in which once-weekly dosing of epoetin beta was as effective as the same total dose administered two or three times weekly. New developments in delivery devices for self-administration, which are more convenient and acceptable to patients, also contribute to the ability to individualize treatment. Furthermore, educating patients in the management of renal anaemia encourages collaboration between patients and health care providers, enabling more tailored treatment regimens to be developed. In conclusion, the use of flexible dosing frequencies, easy-to-use delivery devices and patient education may increase patient compliance and encourage self administration of epoetin beta. The ability to individualize epoetin beta therapy increases the number of options available to nephrologists and their patients, leading to greater opportunities to tailor treatment according to specific needs. PMID- 12091596 TI - A rationale for an individualized administration frequency of epoetin beta: a pharmacological perspective. AB - Several studies have compared the efficacy of once-weekly subcutaneous (s.c.) epoetin treatment with two or three times weekly treatment in renal anaemia. Epoetin administration frequency has attracted a high level of attention in recent years, and numerous small-scale studies have shown comparable efficacy and tolerability of once-weekly vs more frequent administration. The results of two large-scale, randomized, controlled trials of once-weekly administration of epoetin beta became available recently. One of these studies, by Locatelli et al., was the first to be designed specifically to demonstrate therapeutic equivalence between once-weekly and three times weekly epoetin beta treatment, using rigorous statistical methods. This was a large, multicentre, randomized, parallel group, 24-week study in 173 chronic renal failure patients. Treatment regimens were considered equivalent if: (i) the 90% confidence interval (CI) of the difference between treatment groups was within +/-2% for the time-adjusted area under the haematocrit (Hct) curve (AUC); and (ii) for mean weekly epoetin beta dose, the 90% CI of the ratio of the groups was between 0.8 and 1.25. As recommended by current guidelines for statistical analysis of clinical trial data, multiple analysis populations were examined in order to demonstrate robustness of the results with regard to the population chosen for analysis. Findings from the primary analysis, the per-protocol population, were confirmed by both the intent-to-treat analysis and an exploratory analysis that examined the influence of five patients who received dose increases above the mean. In all three analyses, the 90% CIs were within the pre-specified equivalence ranges for both the difference between treatment groups for Hct AUC and the ratio of mean weekly epoetin beta dose. In conclusion, once-weekly and three times weekly s.c. epoetin beta treatment regimens are statistically equivalent in terms of maintaining stable Hct levels and dose requirements in haemodialysis patients. The agreement of the three analysis populations provides a convincing demonstration of the robustness of the results. These results confirm that a once weekly epoetin beta regimen is an effective option for management of renal anaemia that may improve patient convenience and compliance. PMID- 12091597 TI - A need for an individualized approach to end-stage renal disease patients. AB - Evidence suggests that an individualized and flexible approach may be beneficial to end-stage renal disease (ESRD) patients. This article discusses this approach in relation to three issues: target haemoglobin (Hb) level, epoetin dosing frequency/administration and patient management/education programmes. Trial data indicate that each patient's condition should be taken into account when assigning target Hb values. Normalization of Hb is unlikely to be protective in patients with well-established cardiac disease. However, in patients without severe cardiac conditions, normalization is associated with benefits, such as reduction of cardiovascular risk factors and improved quality of life. Data are awaited from trials examining the impact of anaemia correction in patients not yet on renal replacement therapy (RRT). Two large, randomized controlled trials of haemodialysis patients have demonstrated that once-weekly epoetin beta is as effective and as well tolerated as administration two or three times weekly. Additionally, one of these trials showed that once-weekly and three times weekly administrations were equivalent therapeutically in terms of maintaining both stable haematocrit levels and epoetin beta dose requirements. These results suggest that the epoetin beta route and frequency of administration can be individualized according to patient/physician preference. Renal management programmes, which incorporate a multidisciplinary team approach, strategies for early referral of patients and patient education, have an impact on patient outcomes and on RRT modality choice. An individualized programme will help to optimize the use of treatments aimed at delaying the progression of renal failure and its co-morbidities. In conclusion, evidence suggests that an individualized and flexible approach to target Hb values, epoetin beta route and frequency of administration, and patient education/management programmes may be beneficial to patients with ESRD. As early intervention has an impact on patient outcome and the progression of risk factors, this approach may also be appropriate for patients who are not yet receiving RRT. PMID- 12091598 TI - Masterclass in nephrology. Variability of the Extent of Anaemia and Dose Requirements of Erythropoietic Agents in Patients with Renal Failure: Possible Reasons and Therapeutic Approaches. 25-26 January 2002, Berlin, Germany. Proceedings. PMID- 12091599 TI - Is it time for a paradigm shift? Is erythropoietin deficiency still the main cause of renal anaemia? AB - An increasing number of reports documenting resistance to human recombinant erythropoietin (rHuEPO) therapy are challenging the concept that erythropoietin deficiency is the main cause of the anaemia of chronic kidney disease (CKD). In an attempt to establish whether other factors play a more predominant role in the anaemia of CKD, 988 patients receiving dialysis were assessed for a wide range of variables. Data were collected on haematocrit (Hct) levels, rHuEPO dose, dry weight, serum ferritin, transferrin saturation, serum albumin, serum aluminium, serum parathyroid hormone intact, eKt/V for urea, gender, dose of i.v. iron administered, time in hospital, and use of i.v. vancomycin. Hyporesponsiveness to rHuEPO was defined as patients requiring >500 IU/kg/week or failing to achieve Hct levels of >30%. Ninety-two (9.2%) of the 988 patients met the above criteria for hyporesponsiveness to rHuEPO. In 21 of these patients, Hct concentrations remained <30% at 6-month follow-up. There were known haematological causes of refractoriness to rHuEPO in nine of these patients. During extended follow-up, probable causes of hyporesponsiveness were discovered in all but two of the remaining 13 patients. Of 62 dialysis patients who received rHuEPO at doses >500 IU/kg/week, 45 (73%) had Hct concentrations of 33-42%. These patients were responding to the higher doses of rHuEPO with no obvious adverse effects. Lower values of serum ferritin, transferrin saturation, and eKt/V, or higher levels of parathyroid hormone or serum aluminium were not associated with higher rHuEPO dose requirements. These results suggest that erythropoietin deficiency is still the main cause of the anaemia of CKD. Erythropoietin replacement therapy can correct the anaemia in almost all iron replete patients providing enough hormone is given, functional iron deficiency is avoided, aluminium levels and parathyroid toxicities are controlled and that no de novo haematological condition that affects erythropoiesis or red blood cell survival develops. Consideration should be given to modifying the definition of rHuEPO hyporesponsiveness. The US Hct target of 33-36% for haemodialysis patients is narrow and the European target of Hct >33% may be significantly more practical and physiologically relevant. PMID- 12091600 TI - The relationship of haemoglobin level and survival: direct or indirect effects? AB - The relationship between haemoglobin (Hb) level and survival in patients with chronic kidney disease (CKD) is complex. This paper explores the physiological basis for the hypothesis that Hb level and survival are causally related in this patient group, and assesses the current state of knowledge from clinical studies. Issues related to the methodology and analysis of clinical studies limit the certainty with which conclusions regarding the direct relationship between Hb level and survival can be drawn. The data support the concepts that Hb level is associated with survival in patients both with and without CKD, that changes in Hb level are associated with cardiovascular disease (CVD), and that CVD is prevalent in patients with CKD. Hb level is affected by nutritional status, inflammation, and the availability and effectiveness of human recombinant erythropoietin (rHuEPO) therapy, as well as by the degree of kidney function. Thus, the complexity of the relationships between Hb level, CVD and survival in patients with CKD requires further study from both the mechanistic and the clinical perspective. Properly designed clinical trials with survival as an endpoint, as well as data from prospectively measured modifiers of Hb levels and other markers of CVD, are needed to determine the physiological and statistical interaction of these factors in clinical practice. PMID- 12091601 TI - The role of iron in erythropoiesis in the absence and presence of erythropoietin therapy. AB - Preoperative autologous blood donation has served as a model for blood loss anaemia. Studies in these patients, along with clinical trials of i.v. iron and recombinant human erythropoietin (rHuEPO) therapy, have furthered our understanding of the relationship between erythropoietin, iron, and erythropoiesis. With supplemental oral iron, the endogenous erythropoietic response to routine autologous blood donation and to the anaemia of chronic illness has been shown to be modest, but predictable. In more aggressive donation and more severe anaemia, the endogenous erythropoietic response is more substantial, but still predictable. Studies in patients undergoing aggressive phlebotomy whilst receiving rHuEPO demonstrate a wide variation in response to rHuEPO dose. This variability is not related to age or gender and suggests factors such as iron-restricted erythropoiesis may be responsible. Supporting evidence arises from the superior erythropoietic response observed in patients with haemochromatosis. These patients maintain very high serum iron and transferrin saturation levels. In response to serial phlebotomy these patients can mount an endogenous erythropoietin response up to five-times greater than healthy individuals. When treated with rHuEPO, patients with haemochromatosis respond with much greater RBC expansion volumes than patients receiving rHuEPO and iron supplementation. Studies show no difference in the degree of endogenously stimulated erythropoiesis between patients with measurable iron stores and those without. However, when treated with rHuEPO, increased erythropoiesis has been observed in patients with measurable iron stores compared with those without. This suggests that, while oral iron supplementation may be sufficient to keep pace with endogenously stimulated erythropoiesis, it may not be adequate to prevent iron-restricted erythropoiesis during rHuEPO therapy. Some studies have suggested that i.v iron may prevent iron-restricted erythropoiesis during rHuEPO therapy although further research is needed. The availability of better tolerated i.v. iron preparations provides an ideal opportunity to study the value of iron therapy in patients with acute blood loss, particularly those undergoing rHuEPO therapy. PMID- 12091602 TI - Iron and erythropoietin in renal disease. AB - Our knowledge of erythropoiesis and iron in renal disease is limited. The accepted view of the control of erythropoiesis was founded on observations made in a variety of disorders, but the control mechanism in healthy individuals may not be quite the same. Evidence suggests that mechanisms other than erythropoietic stimulation may play a role in increased red blood cell production. Measuring erythropoiesis is complex. The quantitative reticulocyte count is probably the closest practical assessment of erythropoietic activity we can achieve, yet there is very little correlation between circulating erythropoietin level and reticulocyte count in normal and near normal subjects. Oxygen transport in humans depends entirely upon iron. In renal disease, the failure of the erythropoietin positive feedback mechanism can be readily and directly remedied; recombinant human erythropoietin therapy can replace the missing erythropoietin, but this will be negated if iron supply to the erythroid marrow falls short of demand. Measurement of iron stores is also complex. The use of serum ferritin concentration as a direct quantitative estimate of iron in the stores is not advisable, and in practice we have not found the transferrin receptor assay to be useful in identifying patients who require iron therapy. Use of percentage hypochromia as a measure of iron deficiency is complicated by the fact that hypochromic cells are not exclusively a consequence of functional iron deficiency. There are clearly lessons still to be learned in this field and there is much that we do not yet understand about the control of erythropoiesis and iron metabolism in humans. PMID- 12091603 TI - Folate metabolism in renal failure. AB - In most patients with chronic kidney disease, provision of sufficient human recombinant erythropoietin (rHuEPO) and iron replacement therapy will effectively correct renal anaemia. Folate deficiency has been implicated as a contributory factor in renal anaemia and hyporesponsiveness to rHuEPO treatment. As such, the necessity of regular folate supplementation has been debated over the last decade. Although folate loss through dialysis is greater than by urinary excretion, these losses are easily balanced by a normal mixed diet containing 60 g protein/day. Thus, unless patients show significant folate depletion, additional supplementation of folic acid does not appear to have a beneficial effect on erythropoiesis or on responsiveness to rHuEPO therapy. However, a diagnosis of folate deficiency should be considered in patients with chronic renal insufficiency and significant elevation in mean cell volume or hypersegmented polymorphonuclear leucocytes; in patients with malnutrition or a history of alcohol abuse, or in patients hyporesponsive to rHuEPO treatment, especially when accompanied by macrocytosis. Measurements of serum folate are not necessarily indicative of tissue folate stores and red blood cell (RBC) folate measures provide a more accurate picture. Low RBC folate concentrations in these patients indicate the need for folate supplementation. Folate supplementation can also reduce elevated levels of homocysteine in dialysis patients, which may contribute to the high cardiovascular morbidity prevalent in these individuals. High-dose folate therapy (5-15 mg/day) has been shown to reduce plasma homocysteine levels by 25-30% and appears to be well tolerated provided the patient has adequate vitamin B(12) stores. Although long-term benefits of this intervention for cardiovascular protection and patient survival have yet to be established, folic acid is considered a relatively non-toxic and well-tolerated vitamin. PMID- 12091604 TI - Role of secondary hyperparathyroidism in erythropoietin resistance of chronic renal failure patients. AB - Hyperparathyroidism is usually listed among the possible reasons for impaired response to recombinant human erythropoietin (rHuEPO) in patients with renal disease. However, its relevance in the context of other causes of renal anaemia, and the mechanisms by which it may worsen anaemia, are not entirely clear. Possible pathogenic links between anaemia and parathyroid hormone (PTH) include reduced erythropoiesis due to calcitrol deficiency, and direct or indirect effects of PTH on erythropoietin release, red blood cell (RBC) production, survival, and loss. Studies of these mechanisms have produced disparate results, possibly because secondary hyperparathyroidism may have only a relatively minor role in anaemia that may be masked by the confounding effects of other factors with greater impact. Variations in medical treatment or study methodology may also have affected study results. Severe parathyroid overfunction may contribute to the severity of anaemia in uraemic patients and diminish rHuEPO responsiveness in a minority of patients. However, overall, the importance of hyperparathyroidism appears to be minor compared with other factors such as iron deficiency or inflammation. PMID- 12091605 TI - Anaemia, rHuEPO resistance, and cardiovascular disease in end-stage renal failure; links to inflammation and oxidative stress. AB - End-stage renal disease (ESRD) is characterized by a high mortality rate, derived largely from cardiovascular disease (CVD). In patients with ESRD, high levels of pro-inflammatory cytokines and increased oxidative stress are common features that may contribute to malnutrition, anaemia, recombinant human erythropoietin (rHuEPO) resistance, and atherosclerosis. Inflammation predicts poor outcome in ESRD. It is multifactorial in cause and, while it may reflect the underlying CVD, the acute-phase response may also contribute to both oxidative stress and progressive vascular injury. In patients with ESRD, the acute-phase response may be influenced by a number of factors unrelated to dialysis and perhaps by the dialysis procedure itself. Inflammation and the acute-phase response interact with the haematopoietic system at several levels resulting in reduced erythropoiesis, accelerated destruction of erythrocytes, and blunting of the reactive increase in erythropoietin in response to reduced haemoglobin levels. In patients with ESRD, rHuEPO resistance has been linked with inflammation, the latter of which is often associated with a state of functional iron deficiency. Patients with ESRD are thought to have a reduced capacity to handle oxidative stress. There is recent evidence that a relationship may exist between inflammation and oxidative stress and treatment of anaemia with rHuEPO. However, iron may also generate oxidative stress. Controlled trials are needed before evidence-based recommendations for the management of inflammation-induced anaemia and resistance to rHuEPO can be defined. PMID- 12091606 TI - Patient characteristics determining rHuEPO dose requirements. AB - Suboptimal response to recombinant human erythropoietin (rHuEPO) is common in a substantial percentage of patients with chronic kidney disease. Consequently, a higher dose of rHuEPO is needed to attain target haematocrit (Hct) in such patients. Variables that affect rHuEPO dose requirements can be broadly divided into modifiable and immutable characteristics. To date, most of the scientific studies on rHuEPO hyporesponsiveness have focused on modifiable variables that affect rHuEPO response, such as iron status and dialysis adequacy, while exploration of immutable variables has received less attention. This review addresses the key immutable variables that have been suggested as potential determinants of rHuEPO dose requirement. Several investigators, on the basis of analysis of large patient databases, have suggested that diabetic individuals, women, and Black patients on haemodialysis (HD) require a higher dose of rHuEPO than their respective counterparts to attain target Hct. It is unclear whether the observed differences in achieved Hct are due to inherent biological differences in responsiveness to rHuEPO or failure of the investigators to account for modifiable variables that affect rHuEPO dose requirement. Protocol studies with specific a priori hypotheses have failed to confirm some of the findings from these large database analyses. Factors, such as nutritional status, pregnancy, duration of end-stage renal disease, and type of HD vascular access can all potentially modulate response to rHuEPO and should also be considered. PMID- 12091607 TI - Antibodies against rHuEPO: native and recombinant. AB - Recombinant human erythropoietin (rHuEPO) has been used successfully to correct the anaemia of chronic renal failure for more than 12 years. During this time, neutralizing anti-erythropoietin antibodies have been reported in only three patients. However, during the last 2 years, 21 rHuEPO-treated patients have been referred to our laboratory because of sudden resistance to the recombinant hormone and a pure red cell aplasia requiring red blood cell transfusion. The clinical pattern presented by these patients suggested that they could have developed neutralizing anti-erythropoietin antibodies. In all cases, there was evidence of the presence of strong neutralizing antibodies. The ability of the patient's sera to neutralize erythropoietin and to inhibit erythroid colony formation from normal bone marrow was tested first. The presence of anti erythropoietin antibodies was then demonstrated directly by immunoprecipitation of radiolabelled erythropoietin using native, deglycosylated, and denaturated erythropoietin. The reasons for anti-erythropoietin antibody production in these patients is unclear, although it is clearly related to the treatment with rHuEPO. A plausible explanation would be a slight modification in the production process leading to some antigenicity of the manufactured hormone. However, this hypothesis remains unproven. Prompt detection of such antibodies appears necessary to limit antibody titre and to speed recovery. Immunosuppressive treatment was followed by disappearance of the antibodies in 16 of the cases. PMID- 12091608 TI - Anaemia of critical illness-- implications for understanding and treating rHuEPO resistance. AB - The prospect of a shortage of blood for transfusions, increasing awareness of the adverse effects of transfusions, and the availability of human recombinant erythropoietin (rHuEPO) have stimulated interest in the pathogenesis of the anaemia of intensive care unit (ICU) patients. As in the anaemia of chronic illness or chronic renal failure (CRF), the anaemia of ICU patients is a multifactorial process. Blood loss, inappropriately low erythropoietin production, reduced red cell lifespan, reduced iron availability, and inhibition of erythropoiesis by cytokines all contribute to the anaemia of critical illness, although the contributions of the various elements differ depending on the disease aetiology. Evidence is accumulating that use of rHuEPO can induce stimulation of erythropoiesis in critical illness, but at doses that are usually several-fold higher than those used to define resistance to rHuEPO in the current guidelines for the management of anaemia in CRF. Available data suggest that these high doses are well tolerated, at least in the short term. These observations, as well as demonstrating the potential benefits of rHuEPO therapy in critically ill patients, have practical implications for non-ICU patients with CRF who do not respond sufficiently to the usual doses of rHuEPO. Although the risk-benefit ratio relationship for very high doses of rHuEPO needs further consideration, demonstration of rHuEPO efficacy in critical illness should result in a re-evaluation of the 'dose-response relationship' for rHuEPO in patients with less acute and severe illness, including CRF patients hyporesponsive to current dosing regimens. PMID- 12091609 TI - Adjunctive therapy in anaemia management. AB - Iron supplementation is essential for adequate response to recombinant human erythropoietin (rHuEPO) or darbepoetin alfa. Oral iron therapy is often ineffective as the quantity of iron absorbed after oral intake may be insufficient to keep pace with the demands of rHuEPO-stimulated erythropoiesis in patients with end-stage renal disease (ESRD). Currently available i.v. iron preparations include dextran, iron gluconate, and iron sucrose. As rare, but serious, adverse reactions to i.v. iron dextran have been reported, alternative preparations may be preferred. Careful monitoring of iron parameters is required to avoid the effects of over-treatment. Renal anaemia and iron therapy are associated with oxidative stress, leading to a shortening of the lifespan of red blood cells (RBC) and resistance to rHuEPO. rHuEPO therapy may also enhance oxidative stress on RBC. Oxidative stress can be attenuated or prevented by supplementation with vitamin E or melatonin. Vitamin E therapy has also been shown to have a rHuEPO-sparing effect. Disturbances of carnitine metabolism may contribute to the development of renal anaemia in ESRD patients. Oral or i.v. L carnitine therapy results in an increase in haematocrit and a significant decrease in rHuEPO requirement in HD patients. As yet, there is no general recommendation for L-carnitine supplementation for ESRD patients with renal anaemia. PMID- 12091610 TI - Optimizing dose and mode of renal replacement therapy in anaemia management. AB - Iron supplementation is probably the most important factor affecting response to treatment with recombinant human erythropoietin (rHuEPO) in patients with renal anaemia. However, the adequacy of dialysis is also significant. Many factors affect the process of dialysis and its effects. The purity of water used to make up the dialysate from concentrate is important. Inhibitors of erythropoiesis including ions and disinfectants may often be present in treated mains water. In addition, microbiological and pyrogenic contamination of the dialysate frequently occurs, sometimes leading to development or aggravation of anaemia in haemodialysis (HD) patients and also causing an immune response via cytokine activation. Inhibitors of erythropoiesis are also present in endogenous blood in patients with impaired renal function. Adequate dialysis is responsible for removing these mainly small, and possibly medium and large inhibitor molecules, thereby improving anaemia and enhancing response to rHuEPO. The biocompatibility and flux of the membrane used in HD may also have an effect. The removal of medium or large inhibitors of erythropoiesis is inefficient with cellulose membranes, but can potentially be achieved by using more permeable, high-flux membranes. However, in patients with adequate dialysis and sufficient iron and vitamin supplementation, the beneficial effects of a switch from standard cellulose to high-flux membranes have yet to be proven conclusively. Another area in which positive results on correction of anaemia have been seen in small studies is in the use of on-line haemodiafiltration, haemofiltration, or sterile dialysate. However, further large, controlled studies are needed to confirm these effects. PMID- 12091614 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.1. Organization of follow-up of transplant patients after the first year. AB - GUIDELINES: A. All renal transplant recipients should undergo regular laboratory check-ups (at least every 2 or 3 months) and regular medical visits as out patients (at least every 4-6 months) after the first year post-transplant. B. All renal transplant recipients should be seen at least once a year in the transplant centre where the transplantation has been performed or referred to a closer transplant centre for a complete annual evaluation. PMID- 12091611 TI - Optimizing the use of erythropoietic agents-- pharmacokinetic and pharmacodynamic considerations. AB - Erythropoietic agents have transformed the management of renal anaemia. Two forms of recombinant human erythropoietin (rHuEPO) have been available since the early 1990s, and more recently, a second-generation erythropoietic agent, darbepoetin alfa, has been granted a licence for treating this condition. The endogenous erythropoietin molecule may contain anything from 4 to 14 sialic acid residues. The various isoforms have different biological potencies in vivo owing to their different metabolic clearance rates. However, there is a continuing debate about those organs potentially involved in the metabolism of the erythropoietin molecule; the kidneys, liver, and bone marrow have all been suggested as possible participants. Darbepoetin alfa contains two extra N-linked glycosylation consensus sequences increasing the potential maximum number of sialic acid residues from 14 up to 22. In vitro, the affinity of darbepoetin alfa for the erythropoietin receptor is less than for the natural ligand, but this is more than compensated for by the increased potency in vivo. One of the most important factors determining the biological activity of erythropoiesis-stimulating agents is the length of time that the serum concentration of the protein remains above the threshold necessary for erythropoiesis. The pharmacokinetic profile of darbepoetin alfa is distinct from that of rHuEPO, the major difference being the much longer elimination half-life and slower clearance in vivo of darbepoetin alfa leading to prolonged erythropoietic activity. Stimulation of erythropoiesis depends both on an ambient circulating level of erythropoietin and on the mechanisms governing the interaction of the hormone with its receptor. Our knowledge regarding the latter is limited and it is difficult to predict the optimum frequency of administration for an erythropoietic agent. In general, rHuEPO is given two or three times weekly. A small number of studies have supported the once-weekly use of rHuEPO. All clinical trials so far conducted on darbepoetin alfa have demonstrated success with once-weekly and once every other week dosing. PMID- 12091617 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.2.1 Differential diagnosis of chronic graft dysfunction. AB - GUIDELINES: A. Any significant deterioration in graft function should be investigated using the appropriate diagnostic tools and, if possible, therapeutic interventions should be initiated. The usual causes of a decline in glomerular filtration rate after the first year include transplant-specific causes such as chronic allograft nephropathy, acute rejection episodes, chronic calcineurin inhibitor nephrotoxicity, transplant renal artery stenosis and ureteric obstruction, as well as immunodeficiency-related causes and non-transplant related causes, such as recurrent or de novo renal diseases and bacterial infections. B. Any new onset and persistent proteinuria of >0.5 g/24 h should be investigated and therapeutic interventions should be initiated. The usual causes include chronic allograft nephropathy and transplant glomerulopathy, and recurrent or de novo glomerulonephritis. PMID- 12091618 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.2.2 Chronic graft dysfunction. Immunological factors (alloimmunity). AB - GUIDELINE: All recipients of an allogeneic kidney graft should take life-long maintenance immunosuppressive medication. Whereas there is no immunological test to diagnose chronic allograft dysfunction, circumstantial evidence suggests that immunological factors play an important role in its pathogenesis. This evidence is based on experimental data, the beneficial effect of sharing HLA antigens between donor and recipient and post-transplantation immunological monitoring studies. PMID- 12091619 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.2.3 Chronic graft dysfunction. Non-alloimmune factors. AB - GUIDELINES. A. Whereas immunological mechanisms dominate in the initiation and propagation of the injury that leads to chronic allograft dysfunction and nephropathy, there is circumstantial evidence that non-immunological factors, such as advanced donor age, hyperfiltration, overweight, delayed graft function, heavy proteinuria, smoking, arterial hypertension, hypercholesterolaemia and hypertriglyceridaemia, play a role as aggravating or progression factors. It is recommended to prevent or, if possible, treat all these factors. B. As arterial hypertension is very frequent among renal transplant patients and associated with increased graft (and patient) loss, it is recommended to aim at a blood pressure less than 130/85 mmHg in renal transplant patients and <125/75 mmHg in recipients with proteinuria >1 g/day. PMID- 12091620 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.2.4. Chronic graft dysfunction. De novo renal disease after transplantation. AB - GUIDELINES: A. Acute pyelonephritis is relatively frequent in the transplanted kidney and carries a risk of septicaemia. The condition should be recognized and the patient should be treated promptly in the hospital. B. After initiation of any drugs known to induce the development of interstitial nephritis in the transplant patient, it is recommended to monitor renal function and abnormalities in order to detect any side effects rapidly. If interstitial nephritis is observed, it is recommended to stop the offending drug, and to initiate appropriate treatment. C. De novo membranous nephropathy should be considered in cases of proteinuria and nephrotic syndrome after transplantation. Viral infection, such as HCV, should be excluded. D. In the case of the development of graft dysfunction in a transplant patient with Alport's syndrome, one should consider additionally the possibility of de novo anti-glomerular basement membrane (anti-GBM) glomerulonephritis. PMID- 12091622 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.2.6. Chronic graft dysfunction. Late recurrence of other diseases. AB - GUIDELINES: A. In the rare case of recurrent lupus nephritis, no particular treatment is recommended. Only in the few patients with clinically evident flare up is a reinforcement of immunosuppression recommended. B. Recurrence of Henoch Schonlein purpura may occur even in the absence of clinical signs and symptoms. The prognosis for the graft may be severe, particularly in adults. C. In the case of recurrent ANCA-associated renal or systemic vasculitis, it is recommended to reinforce the immunosuppression with appropriate agents. D. Since diabetic nephropathy recurs almost invariably after transplantation, strict control of diabetes and hypertension, and the use of ACE inhibitors and/or angiotensin II receptor antagonists are recommended in order to prevent or slow the risk of recurrence. PMID- 12091624 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.3.1 Long-term immunosuppression. Late steroid or cyclosporine withdrawal. AB - GUIDELINES: A. In order to reduce or avoid long-term serious adverse effects of corticosteroids, such as bone fractures, diabetes mellitus, arterial hypertension, osteoporosis and eye complications, steroid withdrawal should be considered. B. Steroid withdrawal is safe only in a proportion of graft recipients and is recommended only in low-risk patients. The efficacy of the remaining immunosuppression should be considered. C. After steroid withdrawal, graft function has to be monitored very carefully because of the risk of a delayed but continuous loss of function due to chronic graft dysfunction. In the case of functional deterioration or dysfunction, steroids should be re administered. D. Cyclosporine withdrawal might be considered in order to ameliorate nephrotoxicity, arterial hypertension, lipid disorders and hypertrichosis. This can be carried out with no significant long-term risk of progressive graft loss. The efficacy of the remaining immunosuppression should be considered. After cyclosporine withdrawal, careful monitoring for acute rejection is recommended. PMID- 12091621 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.2.5. Chronic graft dysfunction. Late recurrence of primary glomerulonephritides. AB - GUIDELINES: A. In the case of recurrent focal and segmental glomerulosclerosis (FSGS), aggressive treatment with high-dose cyclosporine in children, ACE inhibitors and/or Angiotensin II antagonists, plasma exchange or immunoadsorption may result in remission in some patients. B. In the case of recurrent membranous nephropathy (MN), there is no specific treatment. However, control of risk factors, such as hypertension, heavy proteinuria and hyperlipidaemia, and prevention of thrombotic complications are recommended. C. In the case of recurrent membranoproliferative glomerulonephritis (MPGN), there is no specific treatment. However, control of risk factors, such as hypertension, heavy proteinuria and hyperlipidaemia, and prevention of thrombotic complications are recommended. D. In the case of recurrent IgA glomerulonephritis, use of additional steroids is not yet a validated treatment. The control of risk factors, such as hypertension, heavy proteinuria and hyperlipidaemia, is recommended. E. In the rare case of recurrent anti-glomerular basement membrane (anti-GBM) glomerulonephritis with reappearance of anti-GBM antibodies, it is recommended to initiate plasma exchange and to treat with appropriate immunosuppressive agents (e.g. cyclophosphamide). PMID- 12091625 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.3.2. Long-term immunosuppression. Therapy conversion. AB - GUIDELINE: Conversion of immunosuppressive drug therapy is recommended to avoid or reduce drug-specific adverse effects, and is generally safe for long-term graft outcome. PMID- 12091626 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.3.3. Long-term immunosuppression. Toxicity of immunosuppression. AB - GUIDELINES: A. Careful long-term monitoring of graft recipients is mandatory to discover signs of immunosuppressive drug toxicity, in particular nephrotoxicity. B. In the case of a discrepancy between the drug dose and signs of toxicity, then a thorough pharmacokinetic analysis should be performed. C. Cardiovascular, renal and metabolic risks and the risk of de novo malignancy must be considered in a long-term monitoring programme. PMID- 12091628 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.5.1. Cardiovascular risks. Cardiovascular disease after renal transplantation. AB - GUIDELINES: A. Post-transplant cardiovascular disease is very common, an important cause of morbidity and the first cause of mortality in renal transplant recipients. Therefore, detection and early treatment of post-transplant cardiovascular disease are mandatory. B. Specific risk factors for developing post-transplant cardiovascular disease include pre-transplant cardiovascular disease, arterial hypertension, uraemia (graft dysfunction), hyperlipidaemia, diabetes mellitus, smoking and immunosuppressive treatment. These factors should be targeted for intervention. C. Pre-transplant cardiovascular disease is a major risk factor for post-transplant cardiovascular disease. Therefore, prior to transplantation, it is mandatory to detect and treat symptomatic coronary artery disease, heart failure due to valvular failure or cardiomyopathy, and pericardial constriction. This policy should also be followed in asymptomatic diabetic patients. PMID- 12091627 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.3.4. Long-term immunosuppression. Non-compliance. AB - GUIDELINES: A. The detection of non-compliers should be a permanent concern of the transplant team (doctors, nurses and others). B. Because non-compliance is associated with late graft dysfunction and graft loss, it is important to reduce the proportion of non-compliers by implementing specific educational programmes addressing this problem and the importance of immunosuppressive medications. C. Non-compliance starts during the first year and may increase thereafter. Therefore, the specific educational programme should be repeated and adapted to the need of the transplant recipient, with delivery of few but clear messages. PMID- 12091630 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.5.2. Cardiovascular risks. Arterial hypertension. AB - GUIDELINES: A. Arterial hypertension is often present after renal transplantation and is of multifactorial origin. Pre-transplant arterial hypertension, chronic allograft nephropathy and immunosuppressive therapy are the most frequent causes of post-transplant arterial hypertension. Careful monitoring and treatment of high blood pressure are recommended following transplantation. B. Post-transplant arterial hypertension is associated with an increased incidence of cardiovascular disease in renal transplant patients and is an independent risk factor for graft failure. Therefore, blood pressure control (<130/85 mmHg for renal transplant recipients without proteinuria, and <125/75 mmHg for proteinuric patients) is mandatory in these patients. General measures and pharmacological intervention are necessary in many cases. In proteinuric patients, anti-hypertensive and anti proteinuric agents could be used, and stricter blood pressure control is recommended. C. In patients with uncontrolled arterial hypertension and/or renal function deterioration, underlying causes should be excluded, especially transplant renal artery stenosis. PMID- 12091632 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.5.4. Cardiovascular risks. Post transplant diabetes mellitus. AB - GUIDELINES: A. Post-transplant diabetes mellitus (PTDM) should be identified by regular (every 3 months) fasting blood glucose and/or glycated haemoglobin (HbA1c) measurements. PTDM should be treated as appropriate to achieve normoglycaemia. B. Immunosuppressive therapy should be adjusted to reverse or ameliorate PTDM. PMID- 12091631 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.5.3. Cardiovascular risks. Hyperlipidaemia. AB - GUIDELINES: A. Hyperlipidaemia risk profiles should be identified by regular screening (at least once a year) for cholesterol, HDL-cholesterol, LDL cholesterol and triglyceride blood levels in renal transplant patients. B. In renal transplant patients, hyperlipidaemia must be treated in order to keep the cholesterol/lipid levels within recommended limits according to the number of risk factors. C. Management of hyperlipidaemia after renal transplantation should be the same as for the dialysis population, with, in addition, modification of the immunosuppressive protocol when appropriate. D. Patients should be carefully monitored for adverse effects of lipid-lowering agents or interactions with immunosuppressive drugs. PMID- 12091633 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.5.5. Cardiovascular risks. Hyperhomocysteinaemia. AB - GUIDELINE: Based on the present data, it is not recommended to measure homocysteine levels. PMID- 12091634 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.5.6. Cardiovascular risks. Smoking. AB - GUIDELINE: Cigarette smoking is associated with a high frequency of post transplant cardiovascular disease and may adversely influence patient and graft survival. Active measures against tobacco smoking are recommended. PMID- 12091635 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.5.7. Cardiovascular risks. Obesity and weight gain. AB - GUIDELINE: Obesity (BMI >30 kg/m2) and weight gain are associated with increased prevalence of cardiovascular disease after transplantation. Appropriate dietary and lifestyle measures should be recommended to these patients. PMID- 12091636 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.5.8. Cardiovascular risks. Immunosuppressive therapy. AB - GUIDELINE: Immunosuppressive therapies, especially corticosteroids and anticalcineurin inhibitors; contribute to the prevalence of cardiovascular risk factors, such as arterial hypertension, hyperlipidaemia and hyperglycaemia, and this effect is dose dependent. Reduction of the dose, withdrawal and/or switching to another drug could be useful to control these risk factors. PMID- 12091638 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.6.1. Cancer risk after renal transplantation. Post-transplant lymphoproliferative disease (PTLD): prevention and treatment. AB - GUIDELINES: A. In the first year after organ transplantation, recipients are at the greatest risk of developing lymphoproliferative diseases (PTLDs), which are induced most often by Epstein-Barr virus (EBV) infection, and patients should therefore be screened prior to or at the time of transplantation for EBV antibodies. B. In the rare cases (<5%) where the recipient is EBV seronegative, he or she has a 95% likelihood of receiving an organ from an EBV-seropositive donor, which translates into a high risk of primary EBV infection with seroconversion soon after transplantation. In such cases, the recipient should receive a prophylactic antiviral treatment with acyclovir, valacyclovir or ganciclovir, starting at the time of transplant and lasting for at least 3 months. The specific recommendations given for CMV prophylaxis could be applicable in this situation. C. The treatment of PTLD should be based on accurate pathology with extensive cell markers and phenotyping. The treatment modalities are as follows. Reduction of basal immunosuppression in all cases (either maintain only steroids, or decrease by at least 50% the anti-calcineurin drugs and stop other immunosuppressive drugs). In the case of EBV-positive B-cell lymphoma, antiviral treatment with acyclovir, valacyclovir or ganciclovir may be initiated for at least 1 month or according to the blood level of EBV replication when available. In the case of rare lymphomas from the mucosal-associated lymphoid tissue (MALT) with positive Helicobacter pylori, full eradication of H. pylori should be carried out with a validated protocol. Subsequent H. pylori prophylaxis should be implemented to avoid relapse. In the case of CD20-positive lymphomas, treatment with rituximab, a chimeric monoclonal antibody directed against CD20, should be carried out with one i.v. injection per week for 4 weeks. In the case of diffuse lymphomas or improper response to previous treatment, CHOP chemotherapy should be used alone or in combination with rituximab. The CHOP regimen is cyclophosphamide, doxorubicine, vincristine and prednisone. Complete cessation of immunosuppression with or without graft nephrectomy should also be considered. PMID- 12091640 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.6.2. Cancer risk after renal transplantation. Skin cancers: prevention and treatment. AB - GUIDELINES: D. Due to the high prevalence of skin cancers after organ transplantation, it is highly recommended to inform patients about self awareness. E. Primary prevention should include the avoidance of sun exposure, use of protective clothing and use of an effective sunscreen (protection factor >15) for unclothed body parts (head, neck, hands and arms) in order to prevent the occurrence of squamous-cell carcinoma. This is the most frequent skin tumour in transplant recipients, and its preferential location is the head. F. Recipients with pre-malignant skin lesions (warts, epidermodysplasia verruciformis or actinic keratoses) should be referred early to a dermatologist for active treatment and close follow-up. G. All skin cancers should be completely removed by a dermatologist with appropriate techniques, such as electro-desiccation with curettage, cryotherapy or surgical excision. H. Secondary prevention for recipients should include close follow-up by a dermatologist (at least every 6 months), the use of topical retinoids to control actinic keratoses and to diminish squamous-cell carcinoma recurrence, and reduction of immunosuppression whenever possible. I. In recipients with multiple and/or recurrent skin cancers, the use of systemic retinoids, such as low-dose acitretin, could be recommended for months/years, if well tolerated, in addition to further reduction in immunosuppression whenever possible. PMID- 12091641 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.6.3. Cancer risk after renal transplantation. Solid organ cancers: prevention and treatment. AB - GUIDELINES: J. All renal transplant recipients should have regular ultrasonography of their native kidneys (when applicable) for screening of renal cell carcinomas, which are observed at much higher incidence in both dialysed and transplant patients. K. Guidelines published for screening and prevention of solid organ cancers in the general population should be strictly applied to transplant recipients, who are in general at higher cancer risk, but would benefit equally or even greater. L. All male renal transplant recipients aged 50 and over should have a yearly prostate specific antigen (PSA) test prior to a regular digital rectal examination. M. All female renal transplant recipients should have a yearly cervical (PAP) smear together with regular pelvic examination and regular mammography, according to national recommendations where available. N. All renal transplant recipients should undergo a faecal occult blood testing as a screening for colorectal cancer and other (pre-malignant) lesions, according to national recommendations where available. O. In all these conditions, it is recommended to reduce immunosuppression whenever possible. PMID- 12091642 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.7.1 Late infections. Pneumocystis carinii pneumonia. AB - GUIDELINES: A. Approximately 5% of patients develop Pneumocystis carinii pneumonia (PCP) after renal transplantation if they do not receive prophylaxis. PCP is a severe disease, with a very high fatality rate. Therefore, all renal transplant recipients should receive PCP prophylaxis. The treatment of choice is trimethoprim-sulfamethoxazole (TMP-SMX), at a dose of 80/400 mg/day or 160/800 mg every other day, for at least 4 months. Patients who are treated for rejection should receive TMP-SMX prophylaxis for 3-4 months. B. In the case of TMP-SMX intolerance, aerosolized pentamidine (300 mg once or twice per month) is an alternative for prophylaxis. C. The first-line treatment of PCP is high-dose TMP SMX. Patients with a PaO2 of <70 mmHg initially should be treated parenterally, and the administration of additional steroids should be considered. PMID- 12091644 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.7.2. Late infections. Tuberculosis. AB - GUIDELINES: A. Tuberculosis (TB) is not rare after renal transplantation, and can be life-threatening. Treatment of active TB in renal transplant recipients should be the same as in the general population, i.e. 2 months of quadruple therapy combining rifampin, isoniazid, ethambutol and pyrazinamide, followed by a 4 months double therapy with isoniazid and rifampin. The drug ethambutol should not be used initially if the rate of resistance to isoniazid is less than 4% in the community. B. As rifampin will reduce the plasma concentration of calcineurin antagonists and rapamycin, the blood levels of these agents must be monitored closely. Rifabutin may be used as an alternative to rifampin, as this drug is a less potent inducer of the microsomal P450 enzymes. C. Renal transplant candidates and renal transplant recipients should be screened for latent TB infection. Patients considered to have latent TB infection are defined as: (i) those who display a 5 mm (renal transplant recipients) or a 10 mm (dialysis patients) induration after tuberculin skin testing; (ii) those with chest X-ray images suggestive of past TB infection; (iii) those with a history of past TB infection that was not treated adequately; and (iv) those who have been in close contact with infectious patients. The preferred treatment of latent TB infection is isoniazid 300 mg/day for 9 months. PMID- 12091647 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.9.1. Haematological complications. Anaemia. AB - GUIDELINES: A. Because anaemia is relatively common after kidney transplantation, regular screening and careful evaluation of its causes are recommended. In many cases, post-transplant anaemia is caused by allograft dysfunction. The use of purine synthesis inhibitors (azathioprine and MMF), ACE inhibitors and angiotensin II receptor antagonists may frequently cause post-transplant anaemia. Anaemia is reversible after withdrawing the offending agent. Haemolytic anaemia may develop in transplant recipients. B. Treatment of anaemia should follow the European best practice guidelines for treatment of anaemia in chronic renal failure. PMID- 12091645 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.8. Bone disease. AB - GUIDELINES: A. All kidney-transplanted patients should undergo a systematic evaluation of their skeletal status, including pre-transplant history of renal osteodystrophy, history of fractures and plasma concentrations of calciotropic hormones and other parameters, and if possible measurement of bone mineral density (BMD). B. Glucocorticoid therapy should be given at the lowest possible dosage. As long as patients are receiving steroids, vitamin D treatment (ergocalciferol or 1,25-dihydroxyvitamin D) is highly recommended. C. Optimal prevention of bone disease by vitamin D treatment, sufficient calcium intake, sex hormone substitution and appropriate use of thiazide diuretics should be considered in all transplant patients. D. In established osteopenia, bisphosphonate treatment should be considered despite limited information in transplant recipients. E. Persistent tertiary hyperparathyroidism should be observed for 1 year after transplantation whenever possible to allow for a spontaneous involution. F. In patients with GFR <50 ml/min after transplantation, uraemic osteodystrophy should be prevented. PMID- 12091648 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.9.2. Haematological complications. Leukopenia. AB - GUIDELINE: Because leukopenia is relatively common after kidney transplantation, regular screening and careful evaluation of its causes are recommended. Azathioprine and mycophenolate mofetil may lead to leukopenia. The combination of allopurinol and azathioprine should be avoided. Leukopenia is often associated with viral infections. PMID- 12091649 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.9.3. Haematological complications. Erythrocytosis. AB - GUIDELINE: In the case of erythrocytosis, the first-line treatment should be administration of ACE inhibitors or angiotensin II receptor antagonists. PMID- 12091650 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.10. Pregnancy in renal transplant recipients. AB - GUIDELINES: A. Renal transplantation restores fertility, and successful pregnancies have been reported in renal transplant women. In women with normal graft function, pregnancy usually has no adverse effect on graft function and survival. Therefore, women of childbearing age who consider pregnancy should receive complete information and support from the transplant team. B. Pregnancy could be considered safe about 2 years after transplantation in women with good renal function, without proteinuria, without arterial hypertension, with no evidence of ongoing rejection and with normal allograft ultrasound. C. Pregnancy after transplantation should be considered a high-risk pregnancy and should be monitored by both an obstetrician and the transplant physician. Pregnancy should be diagnosed as early as possible. The principal risks are infection, proteinuria, anaemia, arterial hypertension and acute rejection for the mother, and prematurity and low birth weight for the foetus. D. Pregnant women and transplanted patients are at increased risk of infections, especially bacterial urinary tract infections and acute pyelonephritis of the graft. Urine cultures should be performed monthly and all asymptomatic infections should be treated. Monitoring of viral infections is also recommended. (Evidence level B) E. Acute rejection episodes are uncommon but may occur after delivery. Therefore, immunosuppression should be re-adjusted immediately after delivery. F. Because pre-eclampsia develops in 30% of pregnant patients, especially those with prior arterial transplant hypertension, blood pressure, renal function, proteinuria and weight should be monitored every 2-4 weeks, with more attention during the third trimester. Anti-hypertensive agents should be changed to those tolerated during pregnancy. ACE inhibitors and angiotensin II receptor antagonists are absolutely contra-indicated. G. Immunosuppressive therapy based on cyclosporine or tacrolimus with or without steroids and azathioprine may be continued in renal transplant women during pregnancy. Other drugs, such as mycophenolate mofetil and sirolimus, are not recommended based on current information available. Because of drug transfer into maternal milk, breastfeeding is not recommended. H. Vaginal delivery is recommended, but caesarean section is required in at least 50% of cases. Delivery should occur in a specialized centre. In the puerperium, renal function, proteinuria, blood pressure, cyclosporine/tacrolimus blood levels and fluid balance should be closely monitored. PMID- 12091651 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.11 Paediatrics (specific problems). AB - GUIDELINES: A. Kidney transplantation should be the treatment of choice for end stage renal disease (ESRD) in children (up to 16 years of age). Because the incidence rate of ESRD is very low, approximately 1-2 children per million general population or 4-6 children per million childhood population, kidney transplantation in children should be performed in specialized paediatric centres with multidisciplinary experts, i.e. transplant surgeons, anaesthetists and paediatric nephrologists, and optimally should be supported by psychologists, paediatric nurses and social workers. B. Due to the urgent need for transplantation, children should have priority in the allocation systems. In addition, pre-emptive transplantation from either live or cadaveric donors should be offered to all paediatric transplant candidates whenever possible. These protocols will reduce the time on dialysis, thus limiting the retardation of growth and development. C. Absolute contra-indications to renal transplantation in children are extremely rare but should be respected: uncontrollable malignancy, ABO incompatibility, the presence of a current positive cross-match or multi-organ failure. There are few relative or transient contra-indications: history of cancer (Wilms tumour), viral infection (HIV, HBV, EBV), very young age (<6 months), severe mental retardation and/or additional disabilities. D. In contrast to adult patients, primary renal diseases responsible for ESRD in children are mostly congenital and hereditary disorders (60%). Children with massive vesico-ureteric reflux or permanent urinary infection should undergo nephroureterectomy to avoid the development of sepsis. In children with ESRD not due to any urinary tract malformation, pre-transplant bilateral nephrectomy of the native kidney should be considered in the case of severe arterial hypertension, heavy proteinuria or risk of renal cancer. E. Psychosocial evaluation of future transplant recipients and their parents is necessary in assessing compliance with management of dialysis and after transplantation. Poor compliance worsens the outcome of paediatric renal transplantation. F. Routine childhood vaccination should be completed whenever possible prior to transplantation, in addition to vaccination against hepatitis B and varicella. G. The pharmacokinetics of immunosuppressive drugs often differ between adult and paediatric recipients. Therefore, drug monitoring is mandatory in order to find the correct drug dosage. H. Today the actuarial probability of graft survival at 1 year should exceed 90% in unselected renal transplant children, and the acute rejection rate should be lower than 30%. I. Special attention should be paid to specific risk factors in paediatric transplantation, such as thrombotic complications, EBV and CMV infections, post-transplant lymphoproliferative disease (PTLD) and recurrence of original renal disease, mainly in patients with focal segmental glomerulosclerosis (FSGS) or atypical haemolytic-uraemic syndrome (HUS). PMID- 12091652 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.12. Elderly (specific problems). AB - GUIDELINES: A. Because renal transplantation can extend the duration and quality of life in elderly patients (age 60-70 years) with end-stage renal disease, transplantation should be considered in all patients, particularly if special programmes and preparations are applied. B. In elderly kidney transplant recipients, immunosuppression has to be adapted to avoid both rejections and adverse effects. C. Accurate diagnosis and aggressive treatment of cardiovascular disease in elderly recipients are recommended because of the high number of deaths with functioning grafts. D. The high risk of concomitant diseases, such as diabetes mellitus, bone disease and malignancies, needs special consideration. PMID- 12091653 TI - European best practice guidelines for renal transplantation. Section IV: Long term management of the transplant recipient. IV.13 Analysis of patient and graft survival. AB - GUIDELINES: A. It is important for a transplant unit to follow-up on the results of their transplant activities. In order to achieve correct reports on graft and patient outcome in all patients, it is necessary to have sufficient resources, such as a computerized database, and continuous updates of patient information. All data collected should be subjected to validation procedures to ensure completeness and accuracy. B. Improved outcomes following implementation of new protocols, based on evaluation of clinical multi-centre trials, should be verified at local transplant centres since centres often include a range of patients different from those selected for the trial. C. The most widely accepted descriptor of outcome is the Kaplan-Meier probability estimate of patient and graft survival. Survival estimates should be calculated at intervals of time after transplantation and should always be expressed with their 95% confidence intervals. D. Kaplan-Meier survival estimates may be calculated in three ways. (i) 'Patient survival' should be calculated from the date of transplantation to the date of death or the date of the last follow-up. (ii) 'Graft survival' (non censored for death) should be calculated from the date of transplantation to the date of irreversible graft failure signified by return to long-term dialysis (or retransplantation) or the date of the last follow-up during the period when the transplant was still functioning or to the date of death. Here, death with graft function is treated as graft failure. (iii) 'Graft survival censored for death with a functioning graft' (death-censored graft survival) should be calculated from the date of transplantation to the date of irreversible graft failure signified by return to long-term dialysis (or retransplantation) or the date of last follow-up during the period when the transplant was still functioning. In the event of death with a functioning graft, the follow-up period is censored at the date of death. E. The outcome of transplants carried out at a centre should be compared with those achieved across a range of data from centres collated by national and international multi-centre registries. Interpretation of a centre's performance should take into account the number of transplants performed and the prevalence of major risk factors. F. Major risk factors that influence transplant outcome are identifiable by applying multivariate analytical methods to large multi-centre follow-up databases. Although these major risk factors may not be identifiable in individual centre data, they should nonetheless be taken into account in patient management. G. When designing a clinical trial or evaluating data from a recent trial, the expected improvement in graft survival resulting from a reduction in acute rejection may be estimated from a knowledge of the rejection and graft survival rates that existed prior to the introduction of the new therapeutic regimen. H. When designing or evaluating a clinical trial, it is important to analyse the power of the study to verify statistically the difference (in graft survival) that might be expected and its statistical significance. A study resulting in absence of statistically significant differences between two treatment groups with insufficient statistical power to verify a difference at the expected level should not be taken as evidence of absence of a true difference. PMID- 12091654 TI - Academic radiology: sustaining the mission. PMID- 12091655 TI - Automatic structuring of radiology reports: harbinger of a second information revolution in radiology. PMID- 12091657 TI - Acquired gastrointestinal fistulas: classification, etiologies, and imaging evaluation. AB - Fistulas are abnormal communications between two epithelial-lined surfaces. Gastrointestinal fistulas encompass all such connections that involve the alimentary tract, and they can be congenital or acquired in nature. This review focuses on acquired gastrointestinal fistulas. Development of an acquired gastrointestinal fistula can greatly affect patient outcome, yet the clinical manifestations are often protean in nature and the etiology, elusive. Imaging plays an important role in the detection and management of acquired gastrointestinal fistulas. The more routine use of cross-sectional imaging (especially computed tomography and magnetic resonance imaging) has altered the standard sequence of radiologic evaluation for possible fistulas, but fluoroscopic studies remain a valuable complement, especially for confirming and defining the anomalous communications. In this review, a classification scheme for gastrointestinal fistulas is provided, major causes are discussed, and individual fistula types are elaborated with an emphasis on contemporary imaging approaches. PMID- 12091658 TI - CT colonography at different radiation dose levels: feasibility of dose reduction. AB - PURPOSE: To investigate the sensitivity and specificity of polyp detection and the image quality of computed tomographic (CT) colonography at different radiation dose levels and to study effective doses reported in literature on CT colonography. MATERIALS AND METHODS: CT colonography and colonoscopy were performed with 100 mAs in 50 consecutive patients at high risk for colorectal cancer; 50- and 30-mAs CT colonographic examinations were simulated with controlled addition of noise to raw transmission measurements. One radiologist randomly evaluated all original and simulated images for the presence of polyps and scored image quality. Differences in image quality were assessed with the Wilcoxon rank test. Scan protocols from the literature and recent (unpublished) updates were collected. RESULTS: In nine of 10 patients with polyps 5 mm in diameter or larger (sensitivity, 90%) and in seven of 17 patients with polyps smaller than 5 mm, polyps were correctly identified with CT colonography at all dose levels. Specificity for patients without polyps 5 mm or larger was 53%-60% at all dose levels and for patients without any polyps was 26% (at 100 and 50 mAs) and 48% (at 30 mAs). Image quality decreased significantly as the dose level decreased. The median effective doses (supine and prone positions) calculated from protocols reported in the literature and updates were 7.8 and 8.8 mSv, respectively. CONCLUSION: Although image quality decreases significantly at 30 mAs (3.6 mSv), polyp detection remains unimpaired. The median dose for CT colonography at institutions that perform CT colonographic research is currently 8.8 mSv. PMID- 12091659 TI - Prospective evaluation of pancreatic tumors: accuracy of MR imaging with MR cholangiopancreatography and MR angiography. AB - PURPOSE: To prospectively assess accuracy of magnetic resonance (MR) imaging, MR cholangiopancreatography (MRCP), and MR angiography in patients suspected of having pancreatic tumors. MATERIALS AND METHODS: Sixty-six patients suspected of having pancreatic tumors underwent MR imaging (unenhanced and contrast material enhanced MR, MRCP, and contrast-enhanced MR angiography). Two blinded readers prospectively analyzed the images by consensus, and results were correlated with surgery, biopsy, or follow-up findings. Results were tabulated in two-by-two tables. RESULTS: MR assessment of pancreatic lesion status (differentiation of benign vs malignant) resulted in 60 correct diagnoses (accuracy, 91%), and six (10%) false diagnoses. Among histologically proved malignant tumors, MR imaging yielded correct diagnoses in 42 of 44 patients (sensitivity, 95%; 95% CI: 85%, 99%), whereas 18 of 22 patients with benign findings were classified correctly. At MR imaging, findings in four patients with chronic pancreatitis were wrongly categorized as malignant tumors (specificity, 82%; 95% CI: 60%, 95%), and in one patient, a distal common bile duct carcinoma was not detected. In no patient with pancreatic adenocarcinoma was this tumor misdiagnosed as benign. In patients with malignant tumors who underwent resection, local-regional tumor growth and vascular infiltration were accurately classified in 89% and 94%, respectively. MR imaging depicted histologically proved synchronous hepatic metastases in 82%. The positive and negative predictive values for cancer nonresectability were 90% and 83%, respectively, and the accuracy, sensitivity, and specificity were 85%, 69%, and 95%, respectively. CONCLUSION: Unenhanced and contrast-enhanced MR imaging with MRCP and MR angiography offers potential as a noninvasive tool for assessment of patients suspected of having pancreatic tumors. PMID- 12091660 TI - Evaluation of an emergency radiology quality assurance program at a level I trauma center: abdominal and pelvic CT studies. AB - PURPOSE: To evaluate the use of a redundant system in improving quality of care in the trauma setting by examining a subset of our quality assurance program. MATERIALS AND METHODS: Five hundred thirty-one consecutive abdominal and pelvic CT reports obtained in patients with trauma at a level I trauma center from August 22, 1999, to August 21, 2000, were retrospectively reviewed. Each case was initially interpreted by a board-certified or board-eligible radiologist during evaluation in the emergency department and was subsequently reviewed by a subspecialty abdominal imaging radiologist as part of a quality assurance program. Nineteen cases were excluded because available information was incomplete, resulting in 512 cases in the current study. Cases with discordant interpretations were followed up to discern care change. RESULTS: Of the 512 trauma cases, 153 (29.9%) showed discordant readings. Review of patient records demonstrated changes in patient care in 12 (7.8%) cases. Three (2.0%) cases were reviewed from the morbidity and mortality records of the Department of Trauma Surgery as a direct result of misinterpretations. Six (4%) cases involved additional diagnostic imaging for reevaluation; in four of these six cases the quality assurance reader's interpretation was confirmed, while in the other two, the initial interpretations were favored. CONCLUSION: Findings suggest that discordant radiologic interpretations most often do not result in a change in patient care and outcome. The quality assurance program did, however, identify and lead to changes in care in a number of cases by providing clinically important additional findings. PMID- 12091661 TI - Transarterial chemoembolization for unresectable hepatocellular carcinoma: meta analysis of randomized controlled trials. AB - PURPOSE: To review the available evidence of chemoembolization for unresectable hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Computerized bibliographic searches with MEDLINE and CANCERLIT databases from 1980 through 2000 were supplemented with manual searches, with the keywords "hepatocellular carcinoma," "liver cell carcinoma," "randomized controlled trial [RCT]," and "chemoembolization." Studies were included if patients with unresectable HCC were enrolled and if they were RCTs in which chemoembolization was compared with nonactive treatment (five RCTs) or if different transarterial modalities of therapy (13 RCTs) were compared. Data were extracted from each RCT according to the intention-to-treat method. Five of the RCTs with a nonactive treatment arm were combined by using the random-effects model, whereas all 18 RCTs were pooled from meta-regression analysis. RESULTS: Chemoembolization significantly reduced the overall 2-year mortality rate (odds ratio, 0.54; 95% CI: 0.33, 0.89; P =.015) compared with nonactive treatment. Analysis of comparative RCTs helped to predict that overall mortality was significantly lower in patients treated with transarterial embolization (TAE) than in those treated with transarterial chemotherapy (odds ratio, 0.72; 95% CI: 0.53, 0.98; P =.039) and that there is no evidence that transarterial chemoembolization is more effective than TAE (odds ratio, 1.007; 95% CI: 0.79, 1.27; P =.95), which suggests that the addition of an anticancer drug did not improve the therapeutic benefit. CONCLUSION: In patients with unresectable HCC, chemoembolization significantly improved the overall 2 year survival compared with nonactive treatment, but the magnitude of the benefit is relatively small. PMID- 12091662 TI - Contrast material travel times in patients undergoing peripheral MR angiography. AB - PURPOSE: To retrospectively review time-resolved two-dimensional projection magnetic resonance (MR) angiographic data to characterize the passage of gadolinium-based contrast material down the leg arteries in patients undergoing peripheral MR angiography. MATERIALS AND METHODS: Data of 87 consecutive standardized peripheral MR angiographic examinations were retrospectively reviewed to determine contrast material arrival times to the common femoral artery (CFA), the popliteal and tibial arteries, and the corresponding veins. Travel times were correlated with clinical data obtained with retrospective chart review and predictive multiparameter models developed with stepwise linear regression. RESULTS: The mean travel time of contrast material to the CFA was 24 seconds +/- 6 (SD), with additional 5 seconds +/- 2 to reach the popliteal artery and 7 seconds +/- 4 to reach the ankle artery. The mean time window of arterial enhancement was 49 seconds +/- 10 in the pelvis, 45 seconds +/- 10 in the thigh, and 35 seconds +/- 14 in the calf. The travel time to CFA was correlated with aortic aneurysm (r = 0.41; P <.001), increasing age (r = 0.31; P =.003), male sex (r = 0.3; P =.005), myocardial infarction (r = 0.26; P =.016), and type 2 diabetes mellitus (r = -0.22; P =.041). Predicted travel time to CFA was 10.6 seconds, plus 0.143 seconds times patient age, plus 4.8 seconds if aneurysm was present, plus 3.8 seconds if male sex, plus 2.8 seconds if history of myocardial infarction. CONCLUSION: Contrast material injected intravenously traveled rapidly down the peripheral arteries at approximately 6 seconds per station, but a long arterial phase time window allowed bolus-chase peripheral MR angiography to function with slower table stepping. Patient-to-patient variations in contrast material kinetics may be anticipated on the basis of age, sex, and clinical parameters. PMID- 12091663 TI - Peripheral vascular disease: combined 3D bolus chase and dynamic 2D MR angiography compared with x-ray angiography for treatment planning. AB - PURPOSE: To compare combined three-dimensional (3D) and two-dimensional (2D) contrast material-enhanced magnetic resonance (MR) angiography with x-ray angiography for planning treatment of peripheral vascular disease. MATERIALS AND METHODS: Three radiologists retrospectively reviewed the pretreatment x-ray angiographic and MR angiographic studies obtained in 30 consecutive patients: 15 patients (15 limbs) evaluated for limb salvage and 15 patients (20 limbs) evaluated because of claudication. MR angiography included acquisition of 2D contrast-enhanced MR digital subtraction angiograms of the area from the adductor canal to the feet and 3D spoiled gradient-recalled-echo bolus chase MR angiograms obtained in three stations from the aorta to the middle portion of the calf. Each reader reviewed the x-ray and MR angiograms to determine the inflow and outflow segments for a hypothetical bypass graft placement. RESULTS: The three readers selected identical segments for inflow at MR angiography and x-ray angiography in 32, 32, and 35 of the 35 limbs evaluated (mean percentages of agreement [95% CI ]: 91% [77%, 98%], 91% [77%, 98%], and 95% [90%, 100%], respectively). The readers selected identical segments for outflow in 32, 32, and 34 of the 35 limbs evaluated (mean percentages of agreement [95% CI]: 91% [77%, 98%], 91% [77%, 98%], and 97% [85%, 100%], respectively). CONCLUSION: Preliminary data support the combining of 2D MR digital subtraction angiography with 3D bolus chase MR angiography to extend the utility of 3D MR angiography in treatment planning to include patients being evaluated for limb salvage, as well as those being evaluated for claudication. PMID- 12091664 TI - Head and neck lymphadenopathy: evaluation with US-guided cutting-needle biopsy. AB - PURPOSE: To evaluate ultrasonography (US)-guided core biopsy in the assessment of 247 patients with cervicofacial lymphadenopathy. MATERIALS AND METHODS: Two hundred sixty US-guided core biopsies were performed in 247 patients with cervicofacial lymphadenopathy. The age of the patients ranged from 1 to 91 years (mean, 50 years). Seventy-four (30%) had a history of malignancy. Biopsies were performed as outpatient procedures with direct US guidance and non-advancing 16 18-gauge core needles. Hospital records were reviewed 6 months to 5 years after biopsy. Final diagnoses were rendered based on results of histologic examination of excised specimens, clinical course, or results of other laboratory studies. RESULTS: Two hundred thirty-eight (92%) core biopsies yielded adequate material. In 28 (11%) patients, the histologic diagnosis was considered highly probable. In the 210 patients in whom adequate material was obtained and an unequivocal histologic diagnosis was given, the sensitivity, specificity, and accuracy of US guided core needle biopsy in differentiating benign from malignant lymphadenopathy were 98.1%, 100%, and 98.7%, respectively. Seventy biopsies were performed in 66 patients with lymphoma. Sensitivity, specificity, and accuracy in differentiating lymphoma from reactive lymphadenopathy were 98.5%, 100%, and 98.7%, respectively. In 53 patients (80%) with lymphoma as a final diagnosis, histologic subclassification was sufficient to guide treatment without the need for surgical biopsy. There were no major complications and only three minor post biopsy hematomas. CONCLUSION: US-guided core biopsy in patients with head and neck lymphadenopathy is a safe outpatient procedure that has a high diagnostic yield and accuracy and frequently obviates surgery. PMID- 12091665 TI - Osteoid osteoma: clinical results with thermocoagulation. AB - PURPOSE: To determine the clinical results in an unselected group of consecutive patients with osteoid osteoma treated with thermocoagulation. MATERIALS AND METHODS: In 97 consecutive patients with clinical and/or radiologic evidence of osteoid osteoma at any location, the clinical symptoms were assessed before and after thermocoagulation with computed tomographic guidance. A good response was defined as disappearance of symptoms that were manifested at presentation and attributed to osteoid osteoma. Clinical assessment was performed prior to discharge; within 2 weeks after the procedure; and at 3, 6, 12, and 24 months follow-up. After 24 months, a postal questionnaire was used for assessment. RESULTS: The mean clinical follow-up after the only or the last thermocoagulation session was 41 months (range, 5-81 months). Response was good after one session of thermocoagulation in 74 (76%) of 97 patients, and the 95% CI was 68% to 85%. Patients with persistent symptoms did well after repeated thermocoagulation (good response in 10 of 12 patients), but results of repeated thermocoagulation were poor in patients with recurrent symptoms (good response in five of 10). The overall success rate after one or two thermocoagulation procedures combined was 92% (89 of 97 patients), and the 95% CI was 86% to 97%. Complications were observed in two patients. CONCLUSION: Percutaneous thermocoagulation is a safe and effective method for treatment of osteoid osteoma at any location. Repeated thermocoagulation is successful in patients with persistent symptoms. PMID- 12091666 TI - Painful metastases involving bone: feasibility of percutaneous CT- and US-guided radio-frequency ablation. AB - PURPOSE: To determine the safety and efficacy of radio-frequency (RF) ablation for pain reduction, quality of life improvement, and analgesics use reduction in patients with skeletal metastases. MATERIALS AND METHODS: Over 10 months, 12 adult patients with a single painful osteolytic metastasis in whom radiation therapy or chemotherapy had failed and who reported severe pain (pain score > or = 4 [scale of 0-10]) over a 24-hour period were treated with percutaneous imaging guided RF ablation with a multi-tined electrode while under general anesthesia. Patient pain was measured with a Brief Pain Inventory 1 day after the procedure, every week for 1 month, and thereafter every other week (total follow-up, 6 months). Patient analgesics use was also recorded at these follow-up intervals. Follow-up contrast material-enhanced computed tomography was performed 1 week after the procedure. Complications were monitored. Analysis of the primary end point was undertaken with paired comparison procedures. RESULTS: Lesion size was 1-11 cm. Before RF ablation, mean worst pain score in a 24-hour period in 12 patients was 8.0 (range, 6-10). At 4 weeks after treatment, mean worst pain decreased to 3.1 (P =.001). Mean pain before treatment was 6.5 and decreased to 1.8 (P <.001) 4 weeks after treatment. Mean pain interference in general activity decreased from 6.6 to 2.7 (P =.002) 4 weeks after treatment. Eight of 10 patients using analgesics reported reduced use at some time after RF ablation. No serious complications were observed. CONCLUSION: RF ablation of painful osteolytic metastases is safe, and the relief of pain is substantial. PMID- 12091667 TI - Imaging of fatty tumors: distinction of lipoma and well-differentiated liposarcoma. AB - PURPOSE: To review the reliability of computed tomographic (CT) and magnetic resonance (MR) imaging features in distinguishing lipoma and well-differentiated liposarcoma. MATERIALS AND METHODS: CT (n= 29) and MR (n = 40) images and radiographs (n = 28) of 60 patients with histologically verified fatty tumors (35 lipomas and 25 well-differentiated liposarcomas) were retrospectively reviewed in 31 females and 29 males (mean age, 56 years; age range, 1-88 years). Images were assessed for adipose tissue content, and non-fatty component was classified (thin and/or thick septa and nodular and/or globular components) as absent, mild, moderate, or pronounced. Also assessed were signal intensity and tissue attenuation of the fatty components and non-adipose elements. RESULTS: Statistically significant imaging features favoring a diagnosis of liposarcoma included lesion larger than 10 cm (P <.001), presence of thick septa (P =.001), presence of globular and/or nodular non-adipose areas (P =.003) or masses (P =.001), and lesion less than 75% fat (P <.001). The most statistically significant radiologic predictors of malignancy were male sex, presence of thick septa, and associated non-adipose masses, which increased the likelihood of malignancy by 13-, nine-, and 32-fold, respectively. Both lipoma and liposarcoma demonstrated thin septa and regions of increased signal intensity on fluid sensitive MR images. CONCLUSION: A significant number of lipomas will have prominent non-adipose areas and will demonstrate an imaging appearance traditionally ascribed to well-differentiated liposarcoma. Features that suggest malignancy include increased patient age, large lesion size, presence of thick septa, presence of nodular and/or globular or non-adipose mass-like areas, and decreased percentage of fat composition. PMID- 12091668 TI - MR arthrography of shoulders after suture-anchor Bankart repair. AB - PURPOSE: To determine the ability of magnetic resonance (MR) arthrography to depict the anatomic reestablishment of the capsulolabral complex after suture anchor Bankart repair. MATERIALS AND METHODS: Thirty patients (25 men, five women; mean age, 28 years) who had undergone suture-anchor Bankart repair of one shoulder underwent MR arthrography before second-look arthroscopy. Ninety-eight anchors were used for the sutures. MR arthrographic diagnosis of anatomic reestablishment of the capsulolabral complex was correlated with arthroscopic findings. Contingency table analysis was performed to determine the relationship between MR arthrographic findings and arthroscopic findings. RESULTS: MR findings of reattachment of the capsulolabral complex were in agreement with arthroscopic findings in 93 anchor points (accuracy, 93 of 98 anchor points; 95%). In 28 shoulders, oblique transverse images obtained with the shoulder in the abduction and external rotation position showed that the anterior band of the inferior glenohumeral ligament (AIGHL) abutted the humeral head and that reattachment of the AIGHL to the glenoid rim was seamless. Arthroscopy revealed satisfactory reestablishment of the capsulolabral complex in these shoulders. In the remaining two shoulders, a pool of contrast material was seen between the AIGHL and humeral head and a "divot" was detected at the point of reattachment of the AIGHL to the glenoid rim. Arthroscopy revealed unsatisfactory reestablishment of the capsulolabral complex. MR arthrographic findings of reattachment of the AIGHL were significantly associated with arthroscopic findings of reestablishment of the capsulolabral complex (P <.01). CONCLUSION: MR arthrography can be reliably used for the postoperative assessment of suture-anchor Bankart repair. PMID- 12091669 TI - Tennis leg: clinical US study of 141 patients and anatomic investigation of four cadavers with MR imaging and US. AB - PURPOSE: To evaluate the ultrasonographic (US) findings in patients with a referring diagnosis of tennis leg and to explore the relative importance of the plantaris tendon and gastrocnemius muscle in the pathogenesis of this condition. MATERIALS AND METHODS: A cadaveric study was performed to outline the normal anatomy of the posterosuperficial compartment of the calf. Magnetic resonance (MR) imaging and US were performed, followed by gross anatomic correlation. US findings in 141 patients referred with a clinical diagnosis of tennis leg were retrospectively reviewed by means of consensus of two radiologists. Images were analyzed with respect to the integrity of the lower-leg musculotendinous units, presence of fluid collection, and deep venous thrombosis. RESULTS: MR imaging and US enabled distinction of the musculotendinous unit of the plantaris from the remaining muscles of the lower extremity in cadaveric specimens. US findings in the 141 patients included rupture of the medial head of the gastrocnemius muscle in 94 patients (66.7%), fluid collection between the aponeuroses of the medial gastrocnemius and soleus muscles without muscle rupture in 30 patients (21.3%), rupture of the plantaris tendon in two patients (1.4%), and partial rupture of the soleus muscle in one patient (0.7%). Deep venous thrombosis was seen in isolation in 14 patients (9.9%). CONCLUSION: In patients with clinical findings of tennis leg who undergo US, abnormalities of the medial gastrocnemius muscle appear to be more common than those of the plantaris tendon. PMID- 12091670 TI - "Missing" sternal ossification center: potential mimicker of disease in young children. AB - PURPOSE: To evaluate the frequency of "missing" sternal ossification center (asynchronous non-ossification) in young children. MATERIALS AND METHODS: Lateral chest radiographs obtained in 229 children (mean age, 3.7 years) were retrospectively evaluated for sternal ossification. Four superior sternal segments were considered normal if they were ossified to a similar degree. A segment was considered asynchronous if decreased ossification, as compared with the remaining sternal segments, was demonstrated or if ossification was absent. Asynchronous ossification of inferior sternal segment 5 was recorded separately. Logistic regression analysis was applied to determine if there was a statistically significant relationship (P <.05) between age or sex and pattern of sternal ossification (normal vs asynchronous). RESULTS: Of the 916 superior four sternal segments (four segments in each of 229 patients) evaluated, 32 (3.5%) showed asynchronously decreased or absent ossification. Locations of these 32 segments follow: segment 1, two (0.2%) instances; segment 2, 14 (1.5%) instances; segment 3, two (0.2%) instances; and segment 4, 14 (1.5%) instances. Inferior segment 5 was not ossified in 73 (31.9%) patients. There was a statistically significant relationship between decreased age and increased likelihood of occurrence of asynchronous ossification of one of the sternal ossification centers 1-4 (P >.003) and of occurrence of asynchronous ossification at sternal segment 2 (P <.018). CONCLUSION: Missing sternal ossification centers occur most commonly at segments 2 and 4. Such asynchronous non-ossifications become less common in older children. PMID- 12091671 TI - Bronchial artery embolization for hemoptysis in young patients with cystic fibrosis. AB - PURPOSE: To review the authors' 15-year experience with bronchial artery embolization (BAE) for treatment of hemoptysis in young patients with cystic fibrosis. MATERIALS AND METHODS: By searching the 1985-1999 radiology database, the authors identified 23 young patients who had been referred to the radiology department for angiography. Twenty of these patients underwent BAE. The 23 medical records were retrospectively reviewed with regard to embolization agents used, embolization success rates, number of repeat embolizations, survival times, and causes of death. RESULTS: BAE was performed on 38 occasions in 20 patients. The mean age of patients at first BAE was 15 years (age range, 7-19 years). The majority (n = 34 [89%]) of BAEs were performed by using polyvinyl alcohol. The immediate success rate after BAE (ie, no recurrent bleeding within 24 hours) was 95% (36 of 38 BAEs). Eleven (55%) patients required more than one BAE, and the median time between first and second embolizations was 4 months (range, 5 days to 61 months). Three patients died as a consequence of severe hemoptysis during induction of anesthesia with intermittent positive pressure ventilation in preparation for BAE. The median survival duration after the first BAE (Kaplan Meier estimate) was 84 months (average follow-up, 61 months; range, 5 days to 169 months). CONCLUSION: BAE had a high success rate for short-term control of bleeding; however, more than half the patients required repeat embolization during the long-term follow-up. PMID- 12091672 TI - Bronchogenic carcinoma after lung transplantation: frequency, clinical characteristics, and imaging findings. AB - PURPOSE: To determine the frequency, clinical characteristics, and radiologic findings of bronchogenic carcinoma in patients surviving more than 1 month after lung transplantation. MATERIALS AND METHODS: The study population was composed of 2,168 consecutive patients at seven lung transplantation centers who survived longer than 1 month after lung transplantation. Medical records, chest radiographs, and computed tomographic (CT) scans obtained at the time of diagnosis and prior images when available were reviewed for various items of information and imaging features. RESULTS: Twenty-four (1%) of the 2,168 patients, all with single-lung transplants, developed cancer in the native lung. Eighteen patients had emphysema, and six had pulmonary fibrosis. The frequencies of cancer in patients with emphysema and fibrosis were 2% (18 of 859 patients) and 4% (six of 147 patients), respectively. Twelve (50%) of their 24 cancers were detected at chest radiography. Fourteen (58%) patients had clinical symptoms. Twenty-one (88%) of the 24 patients had one (n = 11) or more (n = 10) nodules, and nine (38%) had one (n = 8) or more (n = 1) masses visible on CT scans. Nodules and masses were visible on 12 (50%) and seven (29%) of 24 chest radiographs, respectively. Eleven (48%) of 23 cancers for which prior chest radiographs were available were seen retrospectively on prior chest radiographs. CONCLUSION: Bronchogenic carcinoma develops in the native lung of transplant recipients with emphysema and pulmonary fibrosis with frequencies of 2% and 4%, respectively. The carcinomas most commonly manifest as a pulmonary nodule or mass on chest radiographs, with more nodules seen on CT scans. PMID- 12091673 TI - Detection of lung cancer on chest radiographs: analysis on the basis of size and extent of ground-glass opacity at thin-section CT. AB - PURPOSE: To evaluate the detection of small peripheral lung tumors on chest radiographs on the basis of the size of the tumor and its extent of ground-glass opacity (GGO) at thin-section computed tomography (CT). MATERIALS AND METHODS: Chest radiographs of 75 patients with peripheral carcinomas 20 mm in diameter or smaller (26 localized bronchioloalveolar carcinomas [BACs], 49 other carcinomas) and 60 normal chest radiographs were retrospectively reviewed individually by 10 radiologists. The extent of GGO within the lesions at thin-section CT was reviewed retrospectively. The detection rates for localized BAC and other carcinomas on chest radiographs were calculated and were correlated with tumor size and extent of GGO. RESULTS: The mean sensitivity for detection of small peripheral carcinomas was 58.5% +/- 8.8 (standard error) for localized BAC and was 78.6% +/- 5.1 for other carcinomas (P =.024). Lesions that were smaller than 15 mm in diameter and had an extent of GGO of 70% or greater at thin-section CT were more difficult to detect than tumors that had larger diameters or less extensive GGO (chi(2) = 8.13, df = 1, P =.004). CONCLUSION: The detection of small peripheral carcinomas on chest radiographs is influenced by tumor size and extent of GGO as seen at thin-section CT. PMID- 12091674 TI - Improved detection of lung nodules by using a temporal subtraction technique. AB - PURPOSE: To evaluate the effect of a temporal subtraction technique for digital chest radiography with regard to the accuracy of detection of lung nodules. MATERIALS AND METHODS: Twenty solitary lung nodules smaller than 30 mm in diameter, including 10 lung cancers and 10 benign nodules, were used. The nodules were grouped subjectively according to their subtlety. For non-nodular cases, 20 nodules without perceptible interval changes were selected. All chest radiographs were obtained by using a computed radiographic system, and temporal subtraction images were produced by using a program developed at the University of Chicago. The effect of the temporal subtraction image was evaluated by using an observer performance study, with use of receiver operating characteristic analysis. RESULTS: Observer performance with temporal subtraction images was substantially improved (A(z) = 0.980 and 0.958), as compared with that without temporal subtraction images (A(z) = 0.920 and 0.825) for the certified radiologists and radiology residents, respectively. The temporal subtraction technique clearly improved diagnostic accuracy for detecting lung nodules, especially subtle cases. CONCLUSION: The temporal subtraction technique is useful for improving detection accuracy for peripheral lung nodules on digital chest radiographs. PMID- 12091675 TI - Recurrent laryngeal nerve palsy in patients with lung cancer: detection with PET CT image fusion -- report of six cases. AB - Positron emission tomography (PET) with fluorodeoxyglucose (FDG) was performed for preoperative staging of lung cancer. In six of 184 patients, there was an intense FDG accumulation in the lower anterior neck. Fusion of PET and computed tomographic images revealed that the focal FDG uptake was localized in the internal laryngeal muscles. This finding was a result of compensatory laryngeal muscle activation caused by contralateral recurrent laryngeal nerve palsy due to direct nerve invasion by lung cancer of the left mediastinum or lung apices. The knowledge of this pitfall is important to avoid false-positive PET results. PMID- 12091676 TI - Use of natural language processing to translate clinical information from a database of 889,921 chest radiographic reports. AB - PURPOSE: To evaluate translation of chest radiographic reports by using natural language processing and to compare the findings with those in the literature. MATERIALS AND METHODS: A natural language processor coded 10 years of narrative chest radiographic reports from an urban academic medical center. Coding for 150 reports was compared with manual coding. Frequencies and co-occurrences of 24 clinical conditions (diseases, abnormalities, and clinical states) were estimated. The ratio of right to left lung mass, association of pleural effusion with other conditions, and frequency of bullet and stab wounds were compared with independent observations. The sensitivity and specificity of the system's pneumothorax coding were compared with those of manual financial coding. RESULTS: The system coded 889,921 reports on 251,186 patients. On the basis of manual coding of 150 reports, the processor's sensitivity (0.81) and specificity (0.99) were comparable to those previously reported for natural language processing and for expert coders. The frequencies of the selected conditions ranged from 0.22 for pleural effusion to 0.0004 for tension pneumothorax. The database confirmed earlier observations that lung cancer occurs in a 3:2 right-to-left ratio. The association of pleural effusion with other conditions mirrored that in the literature. Bullet and stab wounds decreased during 10 years at a rate consistent with crime statistics. A review of pneumothorax cases showed that the database (sensitivity, 1.00; specificity, 0.996) was more accurate than financial discharge coding (sensitivity, 0.17; P =.002; specificity, 0.996; not significant). CONCLUSION: Internal and external validation in this study confirmed the accuracy of natural language processing for translating chest radiographic narrative reports into a large database of information. PMID- 12091678 TI - Case 48: osteogenesis imperfecta of the temporal bone. PMID- 12091679 TI - Single-bolus technique for spiral CT of laryngopharyngeal squamous cell carcinoma: comparison of different contrast material volumes, flow rates, and start delays. AB - PURPOSE: To evaluate different contrast material volumes, flow rates, and start delays for contrast material enhancement of neck structures and squamous cell carcinoma to determine the most effective examination protocol. MATERIALS AND METHODS: Seventy patients with squamous cell carcinoma were prospectively randomized into four groups for examination with different protocols (125 mL of contrast material administered at a flow rate of 2.5 mL/sec, 100 mL at 2.0 mL/sec, 90 mL at 1.5 mL/sec, or 70 mL at 1.0 mL/sec). Dynamic series were performed on the tumors and relevant anatomic structures to obtain time attenuation curves. The protocols were compared (analysis of variance and Tukey Kramer tests) with regard to time and level of maximum tumor enhancement and carotid arterial enhancement of more than 150 HU. One selected protocol was tested in 30 additional routine examinations with start delays of 40 seconds (for laryngeal and/or hypopharyngeal tumors, 3-mm collimation) and 45 seconds (for oropharyngeal tumors, 5-mm collimation). RESULTS: Except for the 70-mL bolus administered at 1.0 mL/sec, the other protocols performed similarly well, yielding comparable maximum tumor enhancement at 52 seconds and later. In spite of a smaller volume of 90 mL, due to the prolonged flow time at 1.5 mL/sec, carotid arterial enhancement of more than 150 HU was prolonged (when compared with that in 100- or 125-mL protocols). As a result of these circumstances, injection of 90 mL at 1.5 mL/sec was considered more effective, providing no significant differences in tumor (P =.39) or carotid arterial (P =.52) enhancement between routine examinations and dynamic series. CONCLUSION: A single bolus of 90 mL administered at 1.5 mL/sec appears to be the most desirable protocol for contrast enhancement. PMID- 12091680 TI - Lymphomas and high-grade astrocytomas: comparison of water diffusibility and histologic characteristics. AB - PURPOSE: To determine if water diffusivity within lymphomas and high-grade astrocytomas correlates with cellularity. MATERIALS AND METHODS: Echo-planar diffusion-weighted magnetic resonance (MR) images obtained in 11 patients with brain lymphomas (19 lesions) and in 17 patients with astrocytomas (19 lesions) were retrospectively reviewed. Regions of interest were drawn on apparent diffusion coefficient (ADC) maps in enhancing tumor. ADC values were normalized by dividing ADC values of tumors by those of normal-appearing regions and expressing the quotient as a ratio. Histologic samples from 11 patients with astrocytomas (11 lesions) and seven patients with lymphoma (seven lesions) were reviewed. Cellularity was measured by calculating the percentage of nuclear area and the percentage of cytoplasmic area and expressing the results as the nuclear to-cytoplasmic (N/C) ratio. The ADC and N/C ratios of both tumor types were compared by using a two-tailed t test. RESULTS: Mean ADC ratio of lymphomas was 1.15 (SD, 0.33; standard error of the mean [SEM], 0.10), and that of high-grade astrocytomas was 1.68 (SD, 0.48; SEM, 0.11) (P <.01). Mean N/C ratio of lymphoma was 1.45 (SD, 0.94; SEM, 0.36), and that of high-grade astrocytomas was 0.24 (SD, 0.18; SEM, 0.05) (P <.01). CONCLUSION: Measurements of water diffusivity and cellularity suggest that higher cellularity contributes to more restricted diffusion. PMID- 12091681 TI - Multiple sclerosis: low-frequency temporal blood oxygen level-dependent fluctuations indicate reduced functional connectivity initial results. AB - PURPOSE: To study the correlation of low-frequency blood oxygenation level dependent (BOLD) fluctuations on magnetic resonance (MR) images obtained of the left- and right-hemisphere primary motor regions in healthy control subjects and patients with multiple sclerosis (MS). MATERIALS AND METHODS: Sixteen healthy volunteers and 20 patients with MS underwent MR imaging with a 1.5-T imager by using a protocol designed to monitor low-frequency BOLD fluctuations. Data for low-frequency BOLD fluctuations were acquired with subjects at rest and during continuous performance of a bilateral finger-tapping task. These data were low pass filtered (<0.08 Hz), and cross correlations of all acquired pixels to a region of interest in the left precentral gyrus were calculated. Confidence levels were calculated from the cross correlations. The fraction of pixels in the right precentral gyrus above a confidence level of 95% for correlation with the left precentral gyrus was calculated for each subject. RESULTS: A plot of the fraction of the right precentral gyrus with high correlation with the left precentral gyrus for the finger-tapping state versus the resting state showed a clear discrimination between patients with MS and control subjects. Compared with control subjects, patients with MS generally had a smaller fraction of the pixels in the right precentral gyrus above the confidence level. This finding indicates that our method results in greater than 60% sensitivity and 100% specificity for discriminating patients with MS from control subjects. No significant correlation was found between clinical measures of MS disease and correlations of low frequency BOLD fluctuations between left and right precentral gyri. CONCLUSION: On the basis of the connectivity measure of low-frequency BOLD fluctuations, patients with MS exhibited lower functional connectivity between right- and left hemisphere primary motor cortices when compared with that in control subjects. PMID- 12091682 TI - Data from a professional society placement service as a measure of the employment market for physicians. AB - PURPOSE: To determine whether data from a professional society placement service- the Professional Bureau of the American College of Radiology--are a valid measure of the employment market. MATERIALS AND METHODS: For the United States from 1990 to 1998, the authors compared three placement service measures-the annual number of job listings, job seekers, and listings per seeker-with two presumably valid measures of the employment market-annual total jobs available (which was ascertained from surveys of hiring) and radiologist median income relative to the all-physician median. For the comparisons, both graphic displays of the data and correlation were used. RESULTS: In graphs, patterns of change were similar. The correlation of job listings, which measure demand, with total jobs, which also measure demand, was 0.84 (P =.04). The correlation of (a) job seekers, a measure of supply, and (b) listings per seeker, which involve both supply and demand, with total jobs was substantial but lower: 0.58 (P =.23) and 0.76 (P =.08), respectively. Correlation of the three placement service measures with relative income, which presumably depends on both supply and demand, was 0.80-0.88 (P <.05 for each measure). CONCLUSION: The statistical significance levels of the correlations and the pattern of findings--namely, stronger correlations among measures of the same aspect of the employment market--indicate that these placement service data are valid and reasonably accurate measures of the employment market. PMID- 12091683 TI - The shading sign. PMID- 12091684 TI - Extraprostatic spread of clinically localized prostate cancer: factors predictive of pT3 tumor and of positive endorectal MR imaging examination results. AB - PURPOSE: To identify the factor(s) most predictive of pT3 tumor and those most predictive of a positive endorectal magnetic resonance (MR) imaging result in patients with clinically localized prostate cancer. MATERIALS AND METHODS: At multivariate analysis, five preoperative clinical parameters-prostate-specific antigen (PSA) level, digital rectal examination (DRE) result, Gleason score and number of involved sextants at transrectal US-guided biopsy, and endorectal MR imaging result-were used to predict pT3 tumor in 336 patients who underwent radical prostatectomy. On the basis of results of the first four examinations, multivariate analysis was performed also to determine predictors of a positive MR imaging study. RESULTS: Significant predictors of pT3 tumor were positive MR imaging result (P < 2 x 10(-8)), more than one sextant involved at biopsy (P < 5 x 10(-5)), and PSA level greater than 10 ng/mL (P < 7 x 10(-3)). Significant predictors of a positive MR imaging result were three or more sextants involved at biopsy (P < 10(-5)), positive DRE result (P < 5 x 10(-3)), and PSA level greater than 10 ng/mL (P < 16 x 10(-3)). In the subgroup of 175 patients who had at least three positive biopsy specimens, the sensitivity of MR imaging was 50% for detection of occult pT3 tumor and 69% for detection of extensive pT3 tumor. The overall specificity of MR imaging was 95%. CONCLUSION: Endorectal MR imaging seems to be indicated in carefully selected patients-specifically, those with three or more positive biopsy specimens, a palpable tumor, and/or a PSA level greater than 10 ng/mL. PMID- 12091685 TI - Mammographic screening of TRAM flap breast reconstructions for detection of nonpalpable recurrent cancer. AB - PURPOSE: To evaluate findings from routine mammographic screenings in patients with transverse rectus abdominis musculocutaneous (TRAM) flap reconstructions. MATERIALS AND METHODS: During a 25-month study period, 214 consecutive screening mammograms in 113 asymptomatic women (mean age, 51 years) with TRAM flap reconstructions were obtained. Mastectomies were performed for cancer in 106 (94%) of the 113 women and for prophylaxis in seven (6%). Prospectively, a Breast Imaging Reporting and Data System (BI-RADS) assessment category 1-5 was assigned to each mammogram. Surgical, medical, pathologic, and radiographic records were retrospectively reviewed. CIs were determined by the normal approximation to the binomial distribution. RESULTS: Seven (3%) of 214 examinations were BI-RADS category 4 or 5. Six (86%) of seven patients underwent biopsy. Two (33%) of these six biopsies demonstrated invasive ductal carcinoma. Cancer detection rate for mammography was 1.9% (two of 106) (95% CI: 0.33%, 7.32%) for women with reconstruction for breast cancer during the 2-year period. One (6%) of 16 BI-RADS category 3 examinations later proved to be invasive ductal carcinoma at follow up. No interval cancer was discovered in 171 cases of BI-RADS category 1 or 2 examinations with 1-year follow-up. No cancers occurred in women who underwent prophylactic mastectomy. A biopsy positive predictive value of 33% (95% CI: 6%, 76%) was observed. CONCLUSION: Screening mammography of TRAM flap-reconstructed breasts enables detection of nonpalpable cancer before clinical examination. PMID- 12091686 TI - Breast cancer detection: evaluation of a mass-detection algorithm for computer aided diagnosis -- experience in 263 patients. AB - PURPOSE: To evaluate the performance of a computer-aided diagnosis (CAD) mass detection algorithm in marking preoperative masses. MATERIALS AND METHODS: Digitized mammograms were processed with an adaptive enhancement filter followed by a local border refinement stage. Features were then extracted from each detected structure and used to identify potential masses. The performance of the algorithm was evaluated in independent cases obtained from 263 patients from two institutions. Each case contained one or more pathologically proved breast masses. Contralateral mammograms obtained in the same patients that did not contain a visible lesion were used to estimate the CAD marker rate for the algorithm. The tradeoff between detection sensitivity and the number of CAD marks was analyzed in this study. RESULTS: Malignant masses were detected with the computer in 87% (135 of 156), 83% (130 of 156), and 77% (120 of 156) of the malignant cases at CAD marker rates of 1.5, 1.0, and 0.5 marks per mammogram, respectively. The difference between malignant mass-detection performance in subsets of cases collected at each institution was found to be less than 1%. The detection accuracy for benign masses was lower than that for malignant masses. CONCLUSION: This mass-detection algorithm had a high sensitivity for detection of malignant masses. It may be useful as a second opinion in mammographic interpretation. PMID- 12091687 TI - Transmitted cardiac pulsations as an indicator of transjugular intrahepatic portosystemic shunt function: initial observations. AB - PURPOSE: To determine if cardiac pulsations are visible and quantifiable on spectral waveforms during Doppler ultrasonographic (US) evaluation of transjugular intrahepatic portosystemic shunts (TIPS), and if so, whether their magnitude declines with shunt dysfunction. MATERIALS AND METHODS: Baseline and pre-revision US images obtained in 15 patients with venographically confirmed TIPS malfunction were retrospectively examined for spectral waveform pulsation. Cardiac pulsatility was quantified by using the venous pulsatility index (VPI), the venous equivalent of resistive index. VPIs were obtained at four locations from the main portal vein to the stent-hepatic venous junction. Baseline and follow-up examination results in 11 patients with functional TIPS acted as controls and were evaluated similarly. Baseline and follow-up mean VPIs at all four locations were compared for both sets of patients by using the Newman-Keuls pairwise multiple sample comparison test. The chi(2) test was used to determine if a VPI threshold that would result in an acceptable sensitivity and specificity for shunt dysfunction existed. RESULTS: One hundred twenty mean VPIs were obtained in the study group, and 88 mean VPIs were obtained in the control group. Pre-revision VPIs at each location were significantly lower (P <.01) than all baseline values and than the follow-up values in the control group. A VPI less than 0.16 was 94% sensitive and 87% specific for shunt dysfunction. CONCLUSION: The VPI, a quantitative measure of cardiac pulsation obtained with Doppler US, may be a useful parameter for assessing TIPS function. PMID- 12091688 TI - Right ventricular MR abnormalities in myotonic dystrophy and relationship with intracardiac electrophysiologic test findings: initial results. AB - PURPOSE: To prospectively determine whether a relationship exists between magnetic resonance (MR) imaging abnormalities of the right ventricle (RV) and intracardiac electrophysiologic (EP) test results in patients with myotonic dystrophy. MATERIALS AND METHODS: Conventional T1-weighted single-shot black blood fast spin-echo and gradient-echo MR imaging of the heart was prospectively performed in 32 patients with myotonic dystrophy who required EP testing. Patients were divided into two groups according to EP test results: (a) inducible (n = 15), indicating inducible ventricular tachyarrhythmias, and (b) noninducible (n = 17). Morphologic and functional MR data were analyzed by two independent investigators. Nonparametric statistical methods and kappa statistics were used. RESULTS: No morphologic or functional abnormalities of the RV wall were observed in noninducible patients. Increased signal intensity of the RV wall, indicative of fatty replacement, was identified in 13 inducible patients. Myocardial thinning of the RV was observed in six inducible patients. An overlap of morphologically abnormal areas and areas of hypo- or dyskinesis were present in 11 inducible patients. RV outflow tract diameter was larger and RV ejection fraction was smaller in inducible patients than in noninducible patients, although differences were not significant. Interobserver agreement for MR findings was good (increased signal intensity: kappa = 0.87, P >.30 [pairwise Wilcoxon signed rank test]; myocardial thinning: kappa = 0.87, P >.30; hypo- or dyskinesis: kappa = 1.00, P >.99). There was a strong relationship between MR abnormalities and inducibility during EP testing (increased signal intensity, P <.001; myocardial thinning, P <.01; hypo- or dyskinesis, P <.01). CONCLUSION: The relationship between MR morphologic and functional RV abnormalities and EP testing suggests potential for the use of MR imaging as a noninvasive method to estimate the individual risk of arrhythmia in patients with myotonic dystrophy. PMID- 12091689 TI - Calcium begets calcium: progression of coronary artery calcification in asymptomatic subjects. AB - PURPOSE: To test the hypothesis that the rate of coronary artery calcium progression is sex specific, namely, that it is greater in men than in women, and that it is age related, particularly in women. MATERIALS AND METHODS: This was a retrospective study of the progression of coronary artery calcium in 217 consecutive asymptomatic subjects who underwent at least two electron-beam computed tomographic studies of the heart. Calcium in the distribution of the epicardial arteries was quantified by using both the conventional coronary artery calcium score (CCS) and the calcium volume score (CVS). Linear regression models were used to judge the joint influence of various risk factors, including sex and age, on rates of coronary artery calcium progression. RESULTS: This study included 103 women and 114 men. The mean interval between the subjects' first and last studies was 25 months +/- 11 (SD). Regression analyses clearly demonstrated that the amount of coronary artery calcium present at the initial study was the most important determinant of calcium progression. This was true when coronary artery calcium was quantified by using the conventional CCS (P <.001) or CVS (P <.001). Neither sex nor age was a significant predictor of coronary artery calcium progression. Among traditional risk factors, only hypertension (P =.02) and diabetes (P =.01) were significant independent factors for calcium progression. CONCLUSION: In asymptomatic subjects, the initial CCS and CVS were the most important factors that affected rate of coronary artery calcium progression. Neither age nor sex was as important as these factors in determination of coronary artery calcium progression. PMID- 12091690 TI - Detection of simulated chest lesions by using soft-copy reading: comparison of an amorphous silicon flat-panel-detector system and a storage-phosphor system. AB - PURPOSE: To compare observer performance by using soft-copy images produced by an amorphous silicon flat-panel-detector system and a storage-phosphor system for the detection of simulated chest lesions. MATERIALS AND METHODS: To test the diagnostic performance of these two systems, four types of simulated lesions (nodules, micronodules, lines, and reticular opacities) were superimposed over an anthropomorphic chest phantom. Digital chest radiographs were acquired with amorphous silicon flat-panel-detector (3-K [K = 1,000] matrix, 12 bits) and storage-phosphor radiography (4-K matrix, 10 bits). Six board-certified radiologists evaluated soft-copy images on a high-resolution video monitor (2,048 x 2,560 x 8 bits). A total of 14,400 observations were analyzed in terms of receiver operating characteristics. RESULTS: Average performance in terms of nodule detection was significantly better (P <.05) with the flat-panel-detector system than with the storage-phosphor system. For micronodules, lines, and reticular opacities, no significant detection differences in averaged performance were found between the two detector systems. CONCLUSION: In the evaluation of soft-copy images, the amorphous silicon detector system appears to be superior to the storage-phosphor system for the detection of pulmonary nodules. PMID- 12091691 TI - Bone mineral density measurement with dental quantitative CT prior to dental implant placement in cadaver mandibles: pilot study. AB - PURPOSE: To correlate torque forces during insertion of screw-type dental implants with bone mineral density (BMD) values determined preoperatively. MATERIALS AND METHODS: Dental quantitative computed tomography (CT) was performed with simultaneous imaging of five postmortem mandibles and a calibration standard containing defined concentrations of calcium hydroxyapatite. CT numbers were converted to local BMD values by assuming a linear relationship (BMD = a x HU + b), where a and b are calibration coefficients. The a, b, P, and t values, correlation coefficients, and standard errors were calculated. Dental implants (n = 25) were set, and insertion torques were recorded. BMD was determined at the implantation site and correlated with torque forces recorded during implant insertion. Calibration coefficients derived for specimens were compared with those derived for actual patients. RESULTS: Calibration coefficients (at 120 kV) for the postmortem specimens were a = 0.760 +/- 0.03 (mean +/- SD) and b = 2.8 +/ 3.7 and for the patients were a = 0.804 +/- 0.06 and b = 5.2 +/- 4.2. Calibrated BMD values at the location of dental implants exhibit a significant correlation (R(2) = 0.83, P <.001) with insertion torques on the basis of a second-order model, which yields torque = (0.0055 x BMD + 0.73)(2) for the implants used and the surgical technique applied. CONCLUSION: Correlation exists between BMD measured with dental quantitative CT and the insertion torque of dental implants. PMID- 12091692 TI - Mechanism of parenchymal enhancement of the liver with a microbubble-based US contrast medium: an intravital microscopy study in rats. AB - PURPOSE: To investigate the mechanism of prolonged contrast material enhancement of the liver observed with the lipid-shell ultrasonographic (US) contrast agent AF0150, with use of intravital microscopy. MATERIALS AND METHODS: Eight Sprague Dawley rats were used. Six received fluorescent microspheres to label the Kupffer cells; two were used as controls. The edge of the middle lobe of the liver was transilluminated with white light. Fluorescent microspheres were observed under fluorescence light. After injection of AF0150, behavior of microbubbles was observed for 6 minutes while viewing a single high-power field. Multiple other fields were then assessed for stationary bubbles and their relation to Kupffer cells. The number of bubbles in motion, aggregated, stationary, and associated with labeled cells were counted. RESULTS: Of 590 bubbles, 34 (5.8%) became stationary and 556 (94.2%) kept moving. Of the 34 stationary microbubbles, 21 dislodged within 30 seconds. Microbubbles were homogeneously distributed throughout the lobule, in contrast to the dominant periportal distribution of the labeled Kupffer cells. Among 83 stationary bubbles observed from all fields of view, only 14 (17%) were associated with fluorescent-labeled cells. CONCLUSION: The late parenchymal liver enhancement effect of AF0150 is likely not related to Kupffer-cell uptake, but rather to a mechanical slowdown within the sinusoids. PMID- 12091693 TI - Abdomen: diffusion-weighted MR imaging with pulse-triggered single-shot sequences. AB - Magnetic resonance (MR) diffusion measurements of the abdomen were performed in 12 healthy volunteers by using a diffusion-weighted single-shot sequence both without and with pulse triggering for different trigger delays. Pulse triggering to the diastolic heart phase led to reduced motion artifacts on the diffusion weighted MR images and to significantly improved accuracy and reproducibility of measurements of the apparent diffusion coefficients, or ADCs, of abdominal organs. PMID- 12091694 TI - Microcalcifications in breast tissue phantoms visualized with acoustic resonance coupled with power Doppler US: initial observations. AB - Calcium carbonate particles embedded in gelatin and turkey breast tissues were visualized with acoustic resonance imaging and power Doppler ultrasonography. Sonography revealed that the region of color level detection corresponded to the location of the calcium carbonate particles. Correlation between color level detection and the location of the particles was confirmed on radiographs of the specimens obtained at core needle biopsy performed through the region of color level detection. PMID- 12091695 TI - Whole-body three-dimensional MR angiography with a rolling table platform: initial clinical experience. AB - A technique for performing whole-body magnetic resonance (MR) angiography with multi-station three-dimensional MR angiography by using a self-developed rolling table platform that integrates the surface coil was evaluated in three volunteers and 10 patients. Use of the surface coil resulted in high signal-to-noise and contrast-to-noise ratios, which translated into sensitivity and specificity of 95.3% and 95.2%, respectively, for detection of significant stenoses (luminal narrowing, >50%) in lower extremity peripheral vascular disease. PMID- 12091696 TI - Dementing disorders: volumetric measurement of cerebrospinal fluid to distinguish normal from pathologic findings -- feasibility study. AB - The authors describe a magnetic resonance (MR) imaging technique to quantify the severity and distribution of cerebral atrophy by using automated volumetric analysis of the distribution of cerebrospinal fluid. The MR imaging technique demonstrated high diagnostic sensitivity and specificity in a group of healthy subjects and patients with dementing diseases. The authors conclude that this approach provides valuable clinical information that is complementary to information acquired with standard diagnostic practices. PMID- 12091697 TI - Three-dimensional CT with a modified C-arm image intensifier: feasibility. AB - A portable C arm was modified for cone-beam computed tomography (CT). This three dimensional (3D) CT imaging system facilitated the acquisition of fluoroscopic images during a 190 degrees rotation and computed a 3D data cube (matrix, 256 x 256 x 256; scanning time, 100 seconds) with multiplanar image reformation. The high-contrast resolution, 0.9 line pairs per millimeter, was comparable; the low contrast resolution, minimal; and the radiation dose, 60%-80% lower, as compared with these parameters at spiral CT. The normal anatomy of small joints could be depicted, and the osteosynthesis screws in the talus were correctly identified. PMID- 12091698 TI - Diagnosis and grading of vesicoureteral reflux. PMID- 12091699 TI - Monitoring of biologic effects of focused ultrasound beams on the brain. PMID- 12091700 TI - Core biopsy in men with peritoneal carcinomatosis. PMID- 12091701 TI - Stent placement in femoropopliteal arteries. PMID- 12091702 TI - A look in the wrong place. PMID- 12091708 TI - Polyproline II structure in a sequence of seven alanine residues. AB - A sequence of seven alanine residues-too short to form an alpha-helix and whose side chains do not interact with each other-is a particularly simple model for testing the common description of denatured proteins as structureless random coils. The (3)J(HN alpha) coupling constants of individual alanine residues have been measured from 2 to 56 degrees C by using isotopically labeled samples. The results display a thermal transition between different backbone conformations, which is confirmed by CD spectra. The NMR results suggest that polyproline II is the dominant conformation at 2 degrees C and the content of beta strand is increased by approximately 10% at 55 degrees C relative to that at 2 degrees C. The polyproline II conformation is consistent with recent studies of short alanine peptides, including structure prediction by ab initio quantum mechanics and solution structures for both a blocked alanine dipeptide and an alanine tripeptide. CD and other optical spectroscopies have found structure in longer "random coil" peptides and have implicated polyproline II, which is a major backbone conformation in residues within loop regions of protein structures. Our result suggests that the backbone conformational entropy in alanine peptides is considerably smaller than estimated by the random coil model. New thermodynamic data confirm this suggestion: the entropy loss on alanine helix formation is only 2.2 entropy units per residue. PMID- 12091709 TI - Gene targeting of Gemin2 in mice reveals a correlation between defects in the biogenesis of U snRNPs and motoneuron cell death. AB - Neuronal degeneration in spinal muscular atrophy is caused by reduced expression of the survival motor neuron (SMN) protein. SMN and the tightly interacting Gemin2 form part of a macromolecular complex (SMN complex) that mediates assembly of spliceosomal small nuclear ribonucleoproteins (U snRNPs). We used mouse genetics to investigate the function of this complex in motoneuron maintenance. Reduced Smn/Gemin2 protein levels lead to disturbed U snRNP assembly as indicated by reduced nuclear accumulation of Sm proteins. This finding correlates with enhanced motoneuron degeneration in Gemin2(+/-)/Smn(+/-) mice. Our data provide in vivo evidence that impaired production of U snRNPs contributes to motoneuron degeneration. PMID- 12091710 TI - Defective T cell development and function in calcineurin A beta -deficient mice. AB - The calcium-dependent phosphatase calcineurin and its downstream transcriptional effector nuclear factor of activated T cells (NFAT) are important regulators of inducible gene expression in multiple cell types. In T cells, calcineurin-NFAT signaling represents a critical event for mediating cellular activation and the immune response. The widely used immunosuppressant agents cyclosporin and FK506 are thought to antagonize the immune response by directly inhibiting calcineurin NFAT signal transduction in lymphocytes. To unequivocally establish the importance of calcineurin signaling as a mediator of the immune response, we deleted the gene encoding the predominant calcineurin isoform expressed in lymphocytes, calcineurin A beta (CnA beta). CnA beta(-/-) mice were viable as adults, but displayed defective T cell development characterized by fewer total CD3 cells and reduced CD4 and CD8 single positive cells. Total peripheral T cell numbers were significantly reduced in CnA beta(-/-) mice and were defective in proliferative capacity and IL-2 production in response to PMA/ionomycin and T cell receptor cross-linking. CnA beta(-/-) mice also were permissive to allogeneic tumor-cell transplantation in vivo, similar to cyclosporin-treated wild-type mice. A mechanism for the compromised immune response is suggested by the observation that CnA beta(-/-) T cells are defective in stimulation-induced NFATc1, NFATc2, and NFATc3 activation. These results establish a critical role for CnA beta signaling in regulating T cell development and activation in vivo. PMID- 12091711 TI - Unique fluorescence properties of a cyanobacterium Gloeobacter violaceus PCC 7421: reasons for absence of the long-wavelength PSI Chl a fluorescence at -196 degrees C. AB - We investigated the reason for the absence of the long-wavelength PSI Chl a fluorescence at -196 degrees C in the cyanobacterium Gloeobacter violaceus using two methods: p-nitrothiophenol (p-NTP) treatment and time-resolved fluorescence spectra. The p-NTP treatment showed that PSII Chl a fluorescence was specifically affected in a manner similar to that for Synechocystis sp. PCC 6803 and spinach chloroplasts, although there were no components modified by the p-NTP treatment, indicating an absence of the long-wavelength PSI Chl a fluorescence. The time resolved fluorescence spectra with a time resolution of 1.3 ps and spectral resolution of 1.0 nm gave no indication of the presence of the long-wavelength PSI fluorescence in the wavelength region between 700 nm and 760 nm, indicating that a very fast energy transfer among Chl a molecules could not account for the absence of the long-wavelength PSI fluorescence. From these data, it seems that the absence of the long-wavelength PSI fluorescence is due to a lack of the formation of a component responsible for the fluorescence at -196 degrees C, which may originate from a difference in the amino acid sequence. We discuss the significance of this phenomenon and interpret our findings in terms of the evolution of cyanobacteria. PMID- 12091712 TI - TMBP200, a microtubule bundling polypeptide isolated from telophase tobacco BY-2 cells is a MOR1 homologue. AB - Bundles of microtubules and cross-bridges between microtubules in the bundles have been observed in phragmoplasts, but proteins responsible for forming the cross-bridges have not been identified. We isolated TMBP200, a novel microtubule bundling polypeptide with an estimated relative molecular mass of about 200,000 from telophase tobacco BY-2 cells. Ultrastructural observation of microtubules bundled by purified TMBP200 in vitro revealed that TMBP200 forms cross-bridges between microtubules. The structure of the bundles and lengths of the cross bridges were quite similar to those observed in phragmoplasts, suggesting that TMBP200 participates in the formation of microtubule bundles in phragmoplasts. The cDNA encoding TMBP200 was cloned and the deduced amino acid sequence showed homology to a class of microtubule-associated proteins including Xenopus XMAP215, human TOGp and Arabidopsis MOR1. PMID- 12091714 TI - Physiological significance of the structure and components of the apoplast canal system for water absorption in plants. AB - The non-linear differential equation that describes the coupling between water transport and solute transport in the apoplast canal system in plants was proposed by Katou and Furumoto in 1986. In the present paper, we analytically solved the equation in order to find the law describing the canal system. In the canal system, water transport is regulated linearly by solute transport under physiological conditions. The approximate solution of the differential equations defines the conditions of the structure and components of the apoplast canal for optimal water absorption. Water absorption during cell elongation in plants requires that the apoplast canal be composed of a cell wall with an appropriate diffusion coefficient for solute. PMID- 12091713 TI - Cell-cycle dependent dynamic change of 26S proteasome distribution in tobacco BY 2 cells. AB - The 26S proteasome is known to play pivotal roles in cell-cycle progression in various eukaryotic cells; however, little is known about its role in higher plants. Here we report that the subcellular distribution of the 26S proteasome is dynamically changed in a cell-cycle dependent manner in tobacco BY-2 cells as determined by immunostaining with anti-Rpn10 (a regulatory PA700 subunit) and anti-20S catalytic proteasome antibodies. The 26S proteasome was found to localize not only in nuclear envelopes and mitotic spindles but also in preprophase bands (PPBs) and phragmoplasts appearing in G(2) and M phases, respectively. MG132, a proteasome inhibitor, exclusively caused cell-cycle arrest not only at the metaphase but also the early stage of PPB formation at the G(2) phase and the collapse of the phragmoplast, which seems to be closely related to proteasome distribution in the cells. PMID- 12091715 TI - Suppression of S-adenosylmethionine decarboxylase activity is a major cause for high-temperature inhibition of pollen germination and tube growth in tomato (Lycopersicon esculentum Mill.). AB - Possible involvement of impaired polyamine biosynthesis in the poor performance of tomato pollen (Lycopersicon esculentum Mill.) at high temperatures was investigated. Incubation of pollen at 38 degrees C suppressed the increase of S adenosylmethionine decarboxylase (SAMDC) activity in germinating pollen with little influence on arginine decarboxylase activity. Consequently, spermidine and spermine content in the pollen did not increase at 38 degrees C, while putrescine content increased at both 25 degrees C and 38 degrees C. High-temperature inhibition of pollen germination was alleviated by the addition of spermidine or spermine but not of putrescine to the germination medium. Cycloheximide inhibited SAMDC activity in parallel with pollen germination at 25 degrees C, whereas actinomycin D had no effect on either of them, indicating that enhanced SAMDC activity is associated with de novo protein synthesis. Incubation of crude enzyme extracts at 40 degrees C for 1 h did not affect SAMDC. In addition, high temperatures did not enhance protease activity in germinating pollen. These results indicate that low activity of SAMDC, probably due to impaired protein synthesis or functional enzyme formation, is a major cause for the poor performance of tomato pollen at high temperatures. PMID- 12091716 TI - OSTF1: a HD-GL2 family homeobox gene is developmentally regulated during early embryogenesis in rice. AB - In many eukaryotic organisms, homeobox genes are important regulators that specify the cell fate and body plan in early embryogenesis. In this study, a gene designated OSTF1 (Oryza sativa transcription factor 1) encoding a homeodomain protein in rice was isolated and characterized. The encoded OSTF1, although sharing only approximately 51% sequence identity with other HD-GL2 members, contains four characteristic motifs (an N-terminal acidic region, a homeodomain, a truncated leucine zipper, and a START domain). OSTF1 was detected as a single copy gene in rice. The transcripts were absent in young panicle or mature spikelet before anthesis, but appeared very early in the pollinated grain with a transient profile. In vegetative tissues examined, expression was only detectable in root. In situ hybridization analysis on developing grains revealed that OSTF1 was strongly and uniformly expressed in the embryo at the globular stage and preferentially localized to the protoderm at 3-6 d after pollination. Expression was also detectable in the integument and throughout the endosperm. Although OSTF1 is not closely related to the remaining HD-GL2 members in sequences, this gene exhibits an analogous epidermis-preferential expression pattern. PMID- 12091717 TI - Phytochrome and blue light-mediated stomatal opening in the orchid, paphiopedilum. AB - Guard cells of the orchid genus, Paphiopedilum have been reported to lack developed chloroplasts and detectable chlorophyll a autofluorescence. Paphiopedilum stomata lack a photosynthesis-dependent opening response but have a blue light-specific opening. The present study found that low fluence rate green and red light elicited stomatal opening in Paphiopedilum and this opening was reversed by far red light, indicating the presence of a phytochrome-mediated opening response. Phytochrome-dependent, red light-stimulated opening was largest under low fluence rates and decreased to near zero as fluence rate increased. A recently discovered green light reversibility of blue light-specific stomatal opening was used to probe the properties of the blue light response in Paphiopedilum stomata. Blue light-stimulated opening was completely reversed by green light in the presence of far red light. Red light enhanced the blue light response of Paphiopedilum guard cells when given as a pretreatment or together with blue light. Analysis of guard cell pigments showed that guard cells have small amounts of chlorophyll a and b, zeaxanthin, violaxanthin, antheraxanthin and lutein. Zeaxanthin content increased in response to blue light or ascorbate and declined in the dark or under illumination in the presence of dithiothreitol, indicating the presence of an active xanthophyll cycle. Thus Paphiopedilum stomata possess both a blue light-mediated opening response with characteristics similar to species with normal chloroplast development and a novel phytochrome mediated opening response. PMID- 12091718 TI - Effects of mechanical vibration on seed germination of Arabidopsis thaliana (L.) Heynh. AB - The effects of sinusoidal vibration (40-120 Hz, amplitude equal to or smaller than 0.42 mm) on seed germination of Arabidopsis thaliana were examined. When the amplitude of vibration was fixed at 0.42 mm, vibration with frequencies higher than 70 Hz increased the rate of seed germination. When the frequency of vibration was fixed at 100 Hz, vibration with amplitudes larger than 0.33 mm also increased the rate of germination. The increase in the rate of germination appeared dependent on acceleration calculated from the frequency and amplitude of vibration. Vibration with a maximum acceleration of 70 m s(-2) increased the rate of germination, but the promotive effects leveled off at higher accelerations. Vibration had little effect on seed germination in a starch-deficient mutant, pgm. Thus, the amyloplasts appeared to act as a susceptor that senses mechanical vibrations. No vibration-induced promotion of germination was seen in an ethylene insensitive mutant, etr1, or in the wild type in the presence of aminoethoxyvinylglycine, an inhibitor of ethylene synthesis, suggesting that vibration increased the rate of seed germination through the action of ethylene. PMID- 12091719 TI - Response of rice to Al stress and identification of quantitative trait Loci for Al tolerance. AB - Rice (Oryza sativa L.) shows the highest tolerance to Al toxicity among small grain cereal crops, however, the mechanisms and genetics responsible for its high Al tolerance are not yet well understood. We investigated the response of rice to Al stress using the japonica variety Koshihikari in comparison to the indica variety Kasalath. Koshihikari showed higher tolerance at various Al concentrations than Kasalath. The Al content in root apexes was less in Koshihikari than in Kasalath, suggesting that exclusion mechanisms rather than internal detoxification are acting in Koshihikari. Al-induced secretion of citrate was observed in both Koshihikari and Kasalath, however, it is unlikely to be the mechanism for Al tolerance because there was no significant difference in the amount of citrate secreted between Koshihikari and Kasalath. Quantitative trait loci (QTLs) for Al tolerance were mapped in a population of 183 backcross inbred lines (BILs) derived from a cross of Koshihikari and Kasalath. Three putative QTLs controlling Al tolerance were detected on chromosomes 1, 2 and 6. Kasalath QTL alleles on chromosome 1 and 2 reduced Al tolerance but increased tolerance on chromosome 6. The three QTLs explained about 27% of the phenotypic variation in Al tolerance. The existence of QTLs for Al tolerance was confirmed in substitution lines for corresponding chromosomal segments. PMID- 12091720 TI - CO(2) response element and corresponding trans-acting factor of the promoter for ribulose-1,5-bisphosphate carboxylase/oxygenase genes in Synechococcus sp. PCC7002 found by an improved electrophoretic mobility shift assay. AB - We analyzed the promoter of the genes encoding the ribulose-1,5-bisphosphate carboxylase/oxygenase (rbc) in the cyanobacterium Synechococcus sp. PCC7002 and localized the CO(2)-regulatory element. Cyanobacterial transformants were constructed with several DNA segments of the rbc promoter fused to the chloramphenicol acetyltransferase (CAT) gene, and their acetyltransferase activities were analyzed under 0.03% and 1% CO(2) conditions. We found that the AT-rich element localized from -262 to -291 relative to the rbc translation starting site was required for CO(2)-dependent repression. Fluorescent-labeled oligonucleotide probes of identical sequence to the AT-rich element were reacted with protein extracts from cells cultured under conditions of low and high CO(2) atmospheric content. We detected a gel retardation complex of a strong signal intensity in extracts from cells cultured under 15% CO(2), but only a weak signal from cells cultured under 1% CO(2). Moreover, a DNA affinity precipitation assay identified a 16-kDa protein that bound to nucleotide sequences within the AT-rich element. The partial amino acid sequence of the protein was similar to the deduced protein sequences of ORF129 and ORF155 from Synechocystis 6803. Our findings suggest that the AT-rich element plays a role as a negative CO(2) regulatory element and its trans-acting factor possibly regulates the rbc transcription in response to CO(2) levels. PMID- 12091721 TI - Rpp16 and Rpp17, from a common origin, have different protein characteristics but both genes are predominantly expressed in rice phloem tissues. AB - The genes for two types of rice phloem protein (RPP16 and RPP17) were isolated and characterized. Conservation of five exon sizes as well as splicing positions between the two genes suggest that either RPP16 or RPP17 is a resultant of gene duplication. By protein blot analysis, RPP16 and RPP17 proteins were specifically detected in soluble and insoluble fractions of a crude extract of rice plants, respectively, suggesting that these proteins play different roles in individual cells. The expression of Rpp16 and Rpp17 was monitored by the beta-glucuronidase (gusA) reporter-gene method. Rpp16-gusA and Rpp17-gusA were expressed preferentially in the phloem tissues from different parts of the plant, but almost no GUS staining was observed in the rest of the tissues. In roots of both constructs, interestingly, stronger GUS-accumulation was detected in younger vascular tissues than in aged vascular tissues. In situ hybridization also showed that Rpp17 was more strongly expressed in vascular tissues of tiller buds. These results suggest that transcript of these genes was more abundant in young tissues. The presence of two copies of the gene in higher plants, from a common origin, which have different protein characteristics, indicates that evolutionary diversification might have occurred in the gene function. PMID- 12091722 TI - The APRR3 component of the clock-associated APRR1/TOC1 quintet is phosphorylated by a novel protein kinase belonging to the WNK family, the gene for which is also transcribed rhythmically in Arabidopsis thaliana. AB - In higher plants, clock-controlled circadian rhythms are a longstanding issue in physiology, and a newly emerging paradigm of molecular biology. In the model higher plant Arabidopsis thaliana, several genes have been proposed to encode potential clock-associated components, including a member (APRR1/TOC1) of the pseudo-response regulator family. We previously showed that transcripts of the APRR1/TOC1 family start accumulating after dawn rhythmically and sequentially at approximately 2 h intervals in the order of APRR9-->APRR7-->APRR5-->APRR3- >APRR1/ TOC1. This and other results led us to propose that this APRR1/TOC1 quintet might play coordinate roles in the mechanism underlying circadian rhythms in higher plants. To gain further insight as to such an idea, we here attempt to identify proteins that interact with one of the quintet members, APRR3. The identified component is a novel protein kinase, named WNK1, which is considerably similar to, but clearly distinct from, mitogen-activated protein kinases (MAPKs). It was found that APRR3 is a substrate of this novel protein kinase, the gene for which also shows a rhythmic transcription profile that is well coincident with the APRR3 rhythm. These findings give new insight into the mechanisms underlying the circadian rhythm in A. thaliana, providing a molecular link between the putative clock component, APRR3, and WNK1, a novel protein kinase that might be implicated as a signal transducer. PMID- 12091723 TI - C : N ratio increases in the phloem sap during floral transition of the long-day plants Sinapis alba and Arabidopsis thaliana. AB - In plants of Sinapis alba and Arabidopsis thaliana, leaf exudate (phloem sap) was analysed during and after a single long day inducing flowering and in control short days. The amounts of carbohydrates and amino acids were measured to estimate the organic C : N ratio. In both species, the C : N ratio of the phloem sap increased markedly and early during the inductive treatment, suggesting that an inequality in organic C and N supply to the apical meristem may be important at floral transition. PMID- 12091724 TI - Center for Synchrotron Biosciences' U2B beamline: an international resource for biological infrared spectroscopy. AB - A synchrotron infrared (IR) beamline, U2B, dedicated to the biomedical and biological sciences has been constructed and is in operation at the National Synchrotron Light Source (NSLS) of Brookhaven National Laboratory. The facility is operated by the Center for Synchrotron Biosciences of the Albert Einstein College of Medicine in cooperation with the NSLS. Owing to the broadband nature of the synchrotron beam with brightness 1000 times that of conventional sources, Fourier transform IR spectroscopy experiments are feasible on diffraction-limited sample areas at high signal-to-noise ratios and with relatively short data acquisition times. A number of synchrotron IR microscopy experiments that have been performed in the mid-IR spectral range (500-5000 cm(-1)) are summarized, including time-resolved protein-folding studies in the microsecond time regime, IR imaging of neurons, bone and other biological tissues, as well as imaging of samples of interest in the chemical and environmental sciences. Owing to the high flux output of this beamline in the far-IR region (50-500 cm(-1)), investigations of hydrogen bonding and dynamic molecular motions of biomolecules have been carried out from 10 to 300 K using a custom-made cryostat and an evacuated box. This facility is intended as an international resource for biological IR spectroscopy fully available to outside users based on competitive proposal. PMID- 12091725 TI - Radiation damage of protein crystals at cryogenic temperatures between 40 K and 150 K. AB - X-ray radiation damage of lysozyme single crystals by an intense monochromatic beam from the Advanced Photon Source is studied at cryogenic temperatures between 40 K and 150 K. The results confirm that primary radiation damage is both linearly dependent on the X-ray dose and independent of temperature. The upper limit for the primary radiation damage observed in our previous study [Teng & Moffat (2000), J. Synchrotron Rad. 7, 313-317] holds over the wider temperature range of this study. The X-ray diffraction quality of the data acquired at 40 K is superior to those at 100 K, apparently due to temperature dependence of secondary and tertiary radiation damage and to reduced thermal motion. PMID- 12091726 TI - Structural hierarchy of several proteins observed by wide-angle solution scattering. AB - In the present study using a high-intensity X-ray beam from a third-generation synchrotron radiation source, it is demonstrated that a wide-angle X-ray scattering (WAXS) profile from several globular proteins in solution can reflect not only the overall structures (approximately 300 A distance resolution) but also intramolecular structures ranging to secondary structures (approximately 2.5 A distance resolution). The proteins treated in the present experiments are classified as different types of structure categories, namely, as all-alpha, all beta and alpha + beta proteins. Here the full-range experimental scattering curves are compared with the theoretical curves, suggesting a further availability of the SR-WAXS method for studies of structure hierarchy and the function of proteins in solutions. PMID- 12091727 TI - Temperature- and time-resolved X-ray scattering at thin organic films. AB - Multilayers of an Fe(II)-polyelectrolyte-amphiphile complex (Fe-PAC) were investigated simultaneously by energy-dispersive X-ray reflectivity and in-plane diffraction at the bending-magnet beamline at BESSY II. By recording spectra between room temperature and about 348 K with a time resolution of about 60 s, two phase transitions were identified and the respective activation energies were determined. Owing to a fixed geometrical set-up, an energy-dispersive experiment is suitable for measuring X-ray reflectivity and in-plane X-ray diffraction of thin organic films simultaneously. Installed at a bending magnet at BESSY II and using an energy range of 4 < E < 25 keV, both reflectivity and diffraction spectra can be collected within about 60 s. The performance is demonstrated by temperature- and time-resolved measurements of the phase transition behaviour of multilayers of an Fe-PAC deposited on a silicon support by means of the Langmuir Blodgett (LB) technique. By measuring the X-ray reflectivity while increasing the temperature, it is shown that the original LB phase is not stable and transforms irreversibly into a liquid-crystalline (LC)-like phase at about 318 K. At the same temperature the in-plane diffraction signal vanishes reversibly, reflecting rotational disorder of the hexagonal arrangement of amphiphilic chains. Its activation energy is determined to be about 1.3 eV. At about 338 K a second irreversible phase transition occurs to another LC phase with smaller vertical layer spacing. This transition is reversible between 329 K and 338 K. PMID- 12091728 TI - Diffusion in solids studied by nuclear resonant X-ray and neutron scattering. AB - Nuclear resonant scattering of synchrotron radiation and quasielastic neutron scattering allow the measurement of frequencies, directions and lengths of jumps of diffusing atoms. Both methods have been successfully applied to diffusion in solids. Synergies and respective advantages of these two techniques as well as recent developments are discussed on the basis of an example: diffusion in intermetallic alloys. PMID- 12091729 TI - Identification of copper-based green pigments in Jaume Huguet's Gothic altarpieces by Fourier transform infrared microspectroscopy and synchrotron radiation X-ray diffraction. AB - The scientific investigation of ancient paintings gives a unique insight into ancient painting techniques and their evolution through time and geographic location. This study deals with the identification of the green pigments used by one of the most important Catalan masters in Gothic times, Jaume Huguet. Other pigments and materials have also been characterized by means of conventional techniques such as optical microscopy, scanning electron microscopy and Fourier transform infrared spectroscopy. Synchrotron radiation X-ray diffraction has been used to produce maps of phases at a spatial resolution of 100 microm across chromatic layers. PMID- 12091730 TI - Deterministic retrieval of surface waviness by means of topography with coherent X-rays. AB - A surface profile retrieval technique from multiple X-ray total reflection images taken at various distances with full coherent illumination is demonstrated. An experiment was performed using the 1 km-long BL29XU beamline at the SPring-8 facility, Japan. Obtained results are compared with results from the optical metrology technique (Fizeau's interferometer). Good agreement between X-ray and optical methods proves the validity of the current approach. Meanwhile, the sensitivity of the X-ray technique is several times higher than that of the standard one. This technique is well suited to the needs of characterizing grazing optics for new-generation X-ray sources. PMID- 12091731 TI - Electronic structure of YbXCu(4) (X = In, Cd, Mg) investigated by high-resolution photoemission spectroscopy. AB - The valence-band electronic structure of YbXCu(4) (X = In, Cd, Mg) has been investigated by means of temperature-dependent high-energy-resolution photoemission spectroscopy using a He I resonance line (hnu = 21.22 eV) and synchrotron radiation (hnu = 800 eV). Intensities of the structure due to the Yb(2+) 4f(7/2) states in the He I spectra of YbInCu(4) and YbCdCu(4) gradually increase with decreasing temperature from 300 to 10 K, and Yb(2+) 4f(7/2) structures are clearly observed as peaks near the Fermi level (E(F)) at 10 K. The enhancement of the Yb(2+) 4f(7/2) peak from 50 to 10 K is much greater for YbInCu(4) than for YbCdCu(4). On the other hand, the Yb(2+) 4f(7/2) states of YbMgCu(4) are observed as a broad structure near E(F). In the synchrotron radiation photoemission spectra of YbInCu(4) and YbCdCu(4), the structures due to the Yb(2+) and Yb(3+) 4f states are recognized at all temperatures. The intensity ratio Yb(2+)/Yb(3+) gradually increases with decreasing temperature. The energy separations between the Yb(2+) and Yb(3+) 4f structures of YbInCu(4) increase from 50 to 20 K. For YbMgCu(4), on the other hand, almost only the Yb(2+) structures are observed and little temperature dependence has been detected. PMID- 12091732 TI - High-resolution photoelectron spectroscopy of Heusler-type Fe(2)VAl alloy. AB - The electronic structure of Heusler-type Fe(2)VAl has been studied by high resolution photoelectron spectroscopy with the excitation photon energy hnu ranging from 21.2 eV (the He I laboratory light source) to 904 eV (the soft X-ray synchrotron light source) for clean surfaces prepared by scraping or fracturing polycrystalline and single crystalline specimens. Photoelectron spectra recorded for the fractured surfaces show a 10 eV-wide valence band with fine structures and a clear decrease in the intensity towards the Fermi level E(F), while a high intensity at E(F) and no fine structures are observed for the scraped surface. Comparison with the theoretical density of states (DOS) indicates that the vacuum ultraviolet photoelectron spectra emphasize the transition-metal 3d bands but the soft X-ray photoelectron spectra agree remarkably well with the DOS including the fine structures and the pseudogap at E(F). The present results suggest that the electronic structure of Fe(2)VAl is highly sensitive to possible strain and defects induced by scraping. Bulk electronic structures of Fe(2)VAl are discussed in relation to the reported fascinating transport properties. PMID- 12091733 TI - High-resolution photoemission spectroscopy for the layered antiferromagnetic (La(1-z)Nd(z))(0.46)Sr(0.54)MnO(3). AB - High-resolution HeI photoemission spectroscopy (UPS), Mn 2p-3d resonant photoemission spectroscopy (RPES) and Mn 2p X-ray absorption spectroscopy (XAS) have been performed to investigate the electronic structure and its effect on the electrical resistivity in (La(1-z)Nd(z))(0.46)Sr(0.54)MnO(3) (z = 0, 0, 2, 0.6 and 1.0). It was found that in the UPS and RPES spectra the Fermi edge persisted over the temperature range 15 < or = T < or = 340 K regardless of the magnetic structure or the composition of the samples. The density of states at the Fermi level [N(E(F))] in the samples where 0 < or = z < or = 0.6 was increased drastically at the Curie temperature (T(C)) with decreasing temperature, but essentially kept unchanged across the Neel temperature (T(N)). A fairly large reduction at T(C) and a small increase at T(N) in the electrical resistivity with decreasing temperature are found to be well accounted for in terms of the temperature dependence of N(E(F)). The presence of a finite N(E(F)) in the insulating Nd(0.46)Sr(0.54)MnO(3) was also found. Thus the origin of the insulating behavior in this sample can be regarded as the Anderson localization associated with the small density of states and the chemical disorder between Nd and Sr. PMID- 12091734 TI - Truly bulk-sensitive spectroscopic measurements of valence in heavy fermion materials. AB - Intermediate valence is one of the typical phenomena of systems with strong electronic correlation. The Anderson impurity model predicts a scaling of the valence with the reduced temperature T/T(K), which is difficult to observe by traditional surface-sensitive electronic spectroscopies. This paper presents results obtained by resonant inelastic X-ray scattering (RIXS), a bulk-sensitive configuration- and chemical-specific technique. The temperature dependence of the valence of YbInCu(4) and YbAgCu(4) was measured by tuning the incident energy to the resonance of the Yb(2+) spectral component. In the case of YbInCu(4) a sharp valence transition, as known from thermodynamical measurements, has been found. The valence of YbAgCu(4) reveals a smooth dependence consistent with a Kondo temperature T(K) = 70 K. These findings establish RIXS as a powerful tool for measuring bulk electronic properties of solids. PMID- 12091735 TI - Silicon drift detectors as a tool for time-resolved fluorescence XAFS on low concentrated samples in catalysis. AB - A silicon drift detector (SDD) was used for ex situ and time-resolved in situ fluorescence X-ray absorption fine structure (XAFS) on low-concentrated catalyst samples. For a single-element and a seven-element SDD the energy resolution and the peak-to-background ratio were verified at high count rates, sufficient for fluorescence XAFS. An experimental set-up including the seven-element SDD without any cooling and an in situ cell with gas supply and on-line gas analysis was developed. With this set-up the reduction and oxidation of a zeolite supported catalyst containing 0.3 wt% platinum was followed by fluorescence near-edge scans with a time resolution of 10 min each. From ex situ experiments on low concentrated platinum- and gold-based catalysts fluorescence XAFS scans could be obtained with sufficient statistical quality for a quantitative analysis. Structural information on the gold and platinum particles could be extracted by both the Fourier transforms and the near-edge region of the XAFS spectra. Moreover, it was found that with the seven-element SDD concentrations of the element of interest as low as 100 ppm can be examined by fluorescence XAFS. PMID- 12091736 TI - A new bend-magnet beamline for scanning transmission X-ray microscopy at the Advanced Light Source. AB - The high brightness of the bend magnets at the Advanced Light Source has been exploited to illuminate a scanning transmission X-ray microscope (STXM). This is the first diffraction-limited scanning X-ray microscope to operate with a useful count rate on a synchrotron bend-magnet source. A simple dedicated beamline has been built covering the range of photon energy from 250 eV to 600 eV. The beamline is always available and needs little adjustment. Use of this facility is much easier than that of installations that share undulator beams. This facility provides radiation for C 1s, N 1s and O 1s near-edge X-ray absorption spectromicroscopy with STXM count rates in excess of 1 MHz and with spectral resolution typically 1:2000, limited to about 1:5000. PMID- 12091737 TI - X-ray diffraction from rectangular slits. AB - It is shown that for micrometre-sized beams the X-ray diffraction from slits is a source of strong parasitic background, even for slits of high quality. In order to illustrate this effect, the coherent diffraction from rectangular slits has been studied in detail. A large number of interference fringes with strong visibility have been observed using a single set of slits made of polished cylinders. For very small apertures, asymmetrical slits generate asymmetrical patterns. This pattern is calculated from the theory of electromagnetic field propagation and compared with experiment in the far-field regime. The use of guard slits to remove Fraunhofer diffraction from the beam-defining slits is treated theoretically. Numerical simulations yield the optimum aperture of the guard slits with respect to the distance to the primary slits. Diffraction theory is shown to be essential to understand how to reduce the background-to-signal ratio in high-resolution experiments. PMID- 12091738 TI - Simple scheme for harmonic suppression by undulator segmentation. AB - In order to suppress harmonic intensity of undulator radiation, a scheme similar to the detuning technique used in the double-crystal monochromator is investigated and found to be effective only when the number of periods of the undulator is small, once the finite emittance of the electron beam and angular acceptance of the beamline are taken into account. Instead, a simple scheme is proposed for undulators with many periods: the undulator is divided into several segments and the optical phase in between is adjusted to shift the fundamental energy without significantly affecting other harmonics. PMID- 12091739 TI - Variable linear polarization from an X-ray undulator. AB - A new X-ray undulator has been designed and constructed which produces linearly polarized X-rays in which the plane of polarization can be oriented to a user selectable angle, from horizontal to vertical. Based on the Apple-II elliptically polarizing undulator (EPU), the undulator rotates the angle of the linear polarization by a simple longitudinal motion of the undulator magnets. Combined with the circular and elliptical polarization capabilities of the EPU operating in the standard mode, this new undulator produces soft X-ray radiation with versatile polarization control. This paper describes the magnetic structure of the device and presents an analysis of the magnetic field with varying undulator parameters. The variable linear polarization capability is then exhibited by measuring the X-ray absorption spectrum of an oriented polytetrafluoroethylene thin film. This experiment, which measures the linear dichroism of the sample at two peaks near the C 1s absorption edge, demonstrates the continuous polarization rotation capabilities of the undulator. PMID- 12091743 TI - The use of vocal signals in the social play of barbary macaques. AB - Field studies in various species of Macaca (Cercopithecidae) provided evidence for specific visual displays that typically accompany playful interactions. The aim of our study was to examine whether and when playing individuals would use auditory displays, i.e. vocalizations that often occur during social play as well. The study was conducted on a population of semi-free Barbary macaques (Macaca sylvanus) with a special focus on the composition and dynamics of playful wrestling (synonymous term: 'rough-and-tumble play'). Analyses of dyadic encounters between subadult males allowed us to distinguish five types of playful behaviours and three types of vocalizations. The latter were clearly linked to encounters where effects of visual signals were impaired, e.g. during close body contact. During wrestling, vocalizations tended to increase in the beginning of an encounter, whereas the last seconds of wrestling often showed a decline in vocalization rate. Our results allowed us to conclude that these vocalizations may supplement or in many cases even substitute interactional effects of visual signals, e.g. the 'play face.' PMID- 12091744 TI - Measuring fruit patch size for three sympatric indonesian primate species. AB - Food availability is one of the basic factors affecting primate density and socioecology, but food availability is difficult to assess. Two different ways to obtain accurate estimates of food availability have been proposed: using phenology data or using the behaviour of animals. Phenology data can be refined by only including trees that are large enough to be used; including (potential) tree species in which by the concerned primate species forage; or including (fruiting) trees of these species that actually produce fruit. Alternatively, the sizes of the actually visited trees (foraging trees) give an estimate of fruit availability. These measures are compared for three sympatric primate species at the Ketambe Research Station, Sumatra, Indonesia: the Thomas langur, the long tailed macaque and the orangutan. The sizes of fruiting trees and the foraging trees are larger than the potential trees. The sizes of the potential trees and of the fruiting trees are similar for the three primate species. This, however, is not reflected in the use of trees: the langurs forage on average in trees of similar size to those producing fruit, whereas the macaques and orangutans forage in trees larger than those producing fruit. The use of trees does not necessitate a different cut off point of included dbhs for the three compared primate species. The use of trees of different sizes, however, may be regulated by food competition. This indicates that sympatric primates make different foraging decisions and that behavioural measures of food availability will be less reliable. PMID- 12091745 TI - Fecal testosterone immunoreactivity as a non-invasive index of functional testosterone dynamics in male Japanese macaques (Macaca fuscata). AB - Validation of a simple method for the extraction and quantification of testosterone (T) from the excreta of male Japanese macaques (Macaca fuscata) is presented. Radioimmunoassay of paired fecal and serum samples collected from four intact sexually mature males during the breeding season provided profiles that were significantly correlated when samples were offset by approximately 48 hr. Additionally, no significant differences were observed in the pattern of temporal variation of T levels in serum and feces. Two castrated males were injected with radioinert T, and the patterns of excretion were observed by analysis of serial fecal and urine samples. Approximately 48 hr after the steroid was administered, a significant peak in the average fecal T levels was apparent. The injection event was also registered in the urine of both males, although qualitative differences were observed. These data suggest that measures of fecal T provide a reliable and non-invasive means of assessing gonadal function in this species. As the analysis of hormone levels in feces allows for frequent, stress-free sampling with minimal disruption, this method should be preferred in long-term or in situ applications requiring endocrine monitoring. PMID- 12091746 TI - Grooming site preferences determined by lice infection among Japanese macaques in Arashiyama. AB - I investigated the effect of the density of louse eggs (Pedicinus obtusus and P. eurygaster) on grooming site preferences in Japanese macaques (Macaca fuscata). Louse eggs were more often found on the outer side of the body (upper back, lower back, outer arms, and outer legs) than on the inner side of the body (chest, belly, inner arms, and inner legs). Japanese macaques were more likely to be groomed on the outer side than the inner side of the body by allogrooming and autogrooming. Such grooming site preferences correlated with the distribution of louse eggs but not with the areas of body parts. Thus, the ecology of lice might affect grooming behavior of Japanese macaques. Five hundred and fifty louse eggs were estimated to parasitize an adult female Japanese macaque. Considering the intrinsic rate of natural increase of lice, monkeys need to be groomed almost every day. This suggests that Japanese macaques need grooming partners and form social bonds with others for everyday grooming. PMID- 12091747 TI - Survey of the black howler monkey, alouatta pigra, population at the Mayan site of Palenque, Chiapas, Mexico. AB - A survey of the population of the black howler monkey (Alouatta pigra) present at the Mayan site of Palenque was conducted during 2000. A total of 911 man/hours, spread over 112 days were spent surveying the 600 ha area of pristine forest at the site for howler troops. We detected the presence of 136 individuals of which 131 were members of 20 troops, the rest were 3 solitary adult males and 2 adult males travelling as a pair. Ecological density was estimated at 23 individuals/km(2). Mean troop size was 7.0 individuals and it ranged from 2 - 12 individuals; 60% of the troops were multimale. All sighting of howler monkeys were in evergreen rain forest and 75% were in trees > or = 20 m in height. The reported densities and mean troop size are higher than those reported for the species in Guatemala and in central Quintana Roo, Mexico. The vegetation of the forest contains tree species reported to be used by species of Alouatta in the Moraceae, Sapotaceae, Leguminosae, and Lauraceae plant families. Protection of a large perimeter area (ca 1700 ha) around the archeological site by the Mexican government ensures the conservation of the forest and of the black howler monkey population present at the site. PMID- 12091748 TI - Assessing habitat utilization by neotropical primates: a new approach. AB - This study aims to ascertain habitat utilization, in relation to forest structural variation, by a multi-species group of primates in tropical lowland rainforest in Southeast Peru during dry season. A new approach to assessing habitat utilization was used. Habitat variation was described by structural and indicator variables collected in quadrats along transects through a study area within Terra Firme and Floodplain forest. Variables were grouped into 'factors' accounting for most of the variation by means of a Principal Components Analysis (PCA). Presence or absence of the primates within the quadrats, assessed by repeat transect surveys, was taken to indicate habitat preferences. Discrimination between the habitat and forest structure in areas of primate presence as opposed to absence was carried out by means of Discriminant Function Analysis (DFA). This highlighted patterns in most utilized habitat. Vertical utilization of the forest was also assessed along with presence in bamboo and general activity on encounter. Suggestions of habitat preference and utilization are made for each of the six sympatric species studied, based on significantly discriminating habitat factors, vertical stratification on encounter and relationships with bamboo. Saguinus fuscicollis and Cebus moloch appeared as habitat generalists. Cebus apella, Saimiri sciureus, and Aotus spp., exhibited varying degrees of preference for habitat factors suggesting disturbed forest, Cebus albifrons was more generalistic but with a possible association with primary, naturally disturbed forest. C. apella was encountered in Terra Firme forest significantly more than in Floodplain. For S. sciureus, C. moloch, and C. apella, upper understory was the most utilized forest layer, for C. albifrons, middle canopy and for S. sciureus and Aotus spp., lower understory. Both positive and negative relationships with bamboo were highlighted. Significant positive relationships between Aotus spp., and bamboo suggest dense bamboo stands provide important daytime resting sites for this nocturnal species. C. moloch was also shown to utilize bamboo whereas S. fuscicollis appeared to actively avoid it. High levels of conformity with past studies in terms of habitat utilization patterns described for the species studied supports the conclusion that the methods used in this study provide an effective means of assessing primate habitat utilization within complex habitat. PMID- 12091750 TI - Leaf-grooming by a wild chimpanzee in Mahale. AB - During the course of systematic observations of the leaf-grooming behavior by the chimpanzees (Pan troglodytes) of the Mahale Mountains National Park, Tanzania, I recovered a louse from a leaf "groomed" by an adult male chimpanzee after a typical leaf-grooming session. During the leaf-grooming session I observed a small object on his lower lip. He picked up a leaf, transferred the small object from his lip to the leaf, folded the leaf and crushed the folded side of the leaf with his thumb. I present this observation as further evidence of the "squashing ectoparasites" hypothesis for leaf-grooming. PMID- 12091749 TI - The calming effect of stimuli presentation on infant Japanese Macaques (Macaca fuscata) under stress situation: a preliminary study. AB - In human newborns, presentation of sounds and odors under stress situations had a calming effect on behavioral and cortisol responses (Kawakami et al., 1996, 1997). These calming effects were examined with 10-day- and 15-day-old Japanese macaques. In Study 1, white noise presentation (Experimental conditions) at the blood sampling was compared with no sound presentation (Control conditions) at the blood sampling. White noise presentation has a calming effect on coded behavioral responses. In Study 2, lavender scent presentation (Experimental conditions) at the blood sampling was compared with no odor presentation (Control conditions) at the blood sampling. Lavender presentation has a calming effect on cortisol responses in the marginal level. From the data of these studies, the salivary cortisol levels were related to the cortisol levels in plasma. Our results may suggest the different effects of sound and odor on infant Japanese macaques. PMID- 12091751 TI - Sutton's law and coronary risk factor reduction. PMID- 12091752 TI - Attempts at changing dietary and exercise habits to reduce risk of cardiovascular disease: who's doing what in the community? AB - The objective of this study was to characterize adults in Olmsted County, MN who were attempting to change both their dietary and physical activity habits. A random digit-dial telephone survey was taken of 1232 adults, with questions about lifestyle, medical conditions, demographics, and receipt of previous lifestyle advice from a health care professional. Respondents were grouped in four categories: 1) changing neither diet nor exercise habits (22%); 2) changing diet habits only (20%); 3) changing exercise habits only (9%); and 4) changing both diet and exercise habits (49%). Along with several demographic and behavioral factors, receipt of physician advice to change exercise and dietary habits was a strong predictor of attempts to change both lifestyle habits. Only a minority of the population (23%), however, reported having received such advice. These results support the positive impact of health professional advice on dietary and exercise change in the population. Public health campaigns should be aimed at increasing the provision of such advice. PMID- 12091753 TI - The effect of high-, moderate-, and low-fat diets on weight loss and cardiovascular disease risk factors. AB - Over 60% of Americans are overweight and a number of popular diets have been advocated, often without evidence, to alleviate this public health hazard. This study was designed to investigate the effects of several diets on weight loss, serum lipids, and other cardiovascular disease risk factors. One hundred men and women followed one of four dietary programs for 1 year: a moderate-fat (MF) program without calorie restriction (28 patients); a low-fat (LF) diet (phase I) (16) ; a MF, calorie-controlled (phase II) diet (38 patients); and a high-fat (HF) diet (18 subjects) [corrected]. Weight, total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglycerides (TG), homocysteine (Ho), and lipoprotein(a) [Lp(a)], were measured every 4th month. The TC/HDL-C ratio was calculated and fibrinogen levels were measured at baseline and after one year. The MF diet resulted in a 2.6% (NS) decrease in weight compared with 18.4% (p=0.045) decrease in patients on phase I, 12.6% (p=0.0085) decrease in patients on phase II, and 13.7% (p=0.025) decrease in those on the HF diet. TC was reduced by 5% (NS) in the MF group, 39.1% (p=0.0005) in the phase I group, and 30.4% (p=0.0001) in the phase II group. HF group had a 4.3% (NS) increase in TC. LDL-C was reduced by 6.1% (NS) on MF, 52.0% (p=0.0001) on phase I, and 38.8% (p=0.0001) on phase II. Patients on HF had a 6.0% (NS) increase in LDL-C. There were nonsignificant reductions in HDL-C in those on MF (-1.5%) and HF (-5.8%). Patients on phase I showed an increase in HDL C of 9.0% (NS), while those on phase II diet had a 3.6% increase (NS) in HDL-C. TC/HDL-C increased (9.8%) only in patients following the high-fat diets (NS). Patients on MF had a 5.3% (NS) reduction in TC/HDL-C, while those on LF had significant reductions on the phase I ( -45.8%; p=0.0001) diet and phase II diet (-34.7%; p=0.0001). TG levels increased on both the MF (1.0%) and HF (5.5%) diets, although neither was statistically significant. People following the phase I and II diets showed reductions of 37.3% and 36.9%, respectively. Ho levels increased by 9.7% when people followed the MF diet and by 12.4% when they followed the HF diet. Patients following the phase I and phase II diets showed reductions of 13.6% and 14.6%, respectively. Only those following phase II diets showed a tendency toward significant improvement (p=0.061). Lp(a) levels increased by 4.7% following the MF (NS) diet and by 31.0% (NS) on the HF diet. Patients following phase I showed a 7.4% (NS) reduction and a 10.8% reduction (NS) following phase II. Fibrinogen levels increased only in individuals following HF diets (11.9%), while patients following MF (-0.6%), phase I ( 11.0%), and phase II (-6.3%) diets showed nonsignificant reductions in fibrinogen. Patients on MF demonstrated nonsignificant reductions in weight, LDL C, TC, HDL-C, TC/HDL-C ratios, and fibrinogen and nonsignificant increases in TGs, Lp(a), and homocysteine. There was significant weight loss in patients on phase I and II and HF diets after 1 year. Reductions in TC, LDL-C, TGs, and TC/HDL ratios were significant only in patients either following a LF diet or a MF, calorically reduced diet. Only patients following HF diets showed a worsening of each cardiovascular disease risk factor (LDL-C, TG, TC, HDL-C, TC/HDL ratio, Ho, Lp(a), and fibrinogen), despite achieving statistically significant weight loss. PMID- 12091754 TI - Peripheral arterial disease: medical care and prevention of complications. AB - Peripheral arterial disease (PAD) is a common but under-recognized problem affecting older patients. Intermittent claudication is the most frequent symptom of PAD, although the diagnosis of PAD is often overlooked until the patient presents with limb-threatening ischemia. Importantly, PAD is a marker for generalized atherosclerosis and is closely associated with coronary and cerebrovascular disease. The severity of PAD has been correlated with an increased risk of myocardial infarction, stroke, and cardiovascular death. The recognition and diagnosis of PAD, combined with its appropriate medical management, may well reduce the overall risk of cardiovascular morbidity. When diagnosed early, both exercise and pharmacotherapy can ameliorate symptoms of claudication, augment functional performance, and improve quality of life. PMID- 12091755 TI - The importance of lipid evaluation and management in the prevention and treatment of acute myocardial infarction. AB - There is an obvious need to measure low-density lipoprotein cholesterol in all patients with acute myocardial infarction and coronary artery disease. The recent guidelines of the National Cholesterol Education Program have established the desired level for low-density lipoprotein cholesterol for such patients at <100 mg/dL. However, several studies have demonstrated a lack of low-density lipoprotein cholesterol measurement and lipid-lowering therapy with statins in patients with acute myocardial infarction and coronary artery disease. These findings point to a need for quality of care improvement efforts to foster both lipid measurement and statin use in such patients. Many studies have demonstrated the numerous beneficial effects of statin use. In addition to lipid lowering, these include plaque stability and ant platelet, ant macrophage, and antiatherothrombotic activities as well as enhanced endothelial activity. As a class of drugs, the statins have been shown to offer significant benefits with little in the way of associated risks. PMID- 12091756 TI - Genetics and blood pressure response to exercise, and its interactions with adiposity. AB - Regular aerobic exercise has the potential to induce several beneficial health effects, including a decrease in blood pressure level, especially in hypertensive patients and in subjects with high-normal blood pressure. However, it is also well documented that some people show more pronounced blood pressure responses to endurance training than others, despite identical training programs and similar initial blood pressure levels. This kind of variation is an example of normal biologic diversity and most likely originates from interactions with genetic factors. Data from genetic epidemiologic studies indicate that there is a genetic component that affects both resting blood pressure and blood pressure responses to acute exercise. Evidence from molecular genetic studies is scarce, but the first reports suggest that DNA sequence variation in the hypertension candidate genes, such as angiotensinogen, also modify blood pressure responses to endurance training. The current knowledge regarding the role of genetic factors in the modification of blood pressure responses to endurance training will be summarized and discussed. PMID- 12091757 TI - Intraindividual variability of children's blood lipid and lipoprotein concentrations: a review. AB - Of the numerous risk factors that have been associated with atherosclerosis and subsequent coronary heart disease, none have caught the public's attention more than plasma cholesterol. An increasing number of studies are focusing on intervention strategies aimed at "improving" the plasma lipid-lipoprotein profile of children. However, the efficacy of these strategies cannot be ascertained unless the biologic and analytic variability of these metabolites has been determined. From the limited available literature, it would appear that the biologic variability of total cholesterol, high- and low-density lipoprotein cholesterol, and triacylglycerol in children is similar to that reported for adults. Yet studies that have directly focused on this important issue with children are scarce, especially those that have included a representative sample and measurements of lipoprotein subfractions. Further research is warranted with children to better establish the extent of intraindividual variation associated with the lipid-lipoprotein profile. PMID- 12091758 TI - Perspectives: The NCEP ATP III guidelines-friend or foe? National Cholesterol Education Program. PMID- 12091760 TI - Atrial fibrillation: the newest frontier in arrhythmia management. AB - At least 2.3 million people in the United States have atrial fibrillation. Since the risk for developing atrial fibrillation increases with age, the number of people with atrial fibrillation is expected to rise sharply. Atrial fibrillation is a complex condition that adversely influences mortality, morbidity, quality of life, and use of health care resources. Knowledge generated from extensive research has led to innovative management strategies. As the number of individuals with atrial fibrillation increases and treatment options expand, nurses in a variety of settings will be challenged to respond to the multifaceted needs of this population. This review discusses the significance of atrial fibrillation and summarizes research findings influencing current management strategies. Pharmacologic therapies are reviewed and new technologies for atrial fibrillation treatment are introduced. Nursing assessment and treatment of patients' response to atrial fibrillation are discussed. Recommendations for patient education are offered. A plan describing specific nursing diagnoses, outcomes, interventions, and activities for care of patients with atrial fibrillation is presented. PMID- 12091759 TI - Perspectives: The significance of measuring non-HDL-cholesterol. AB - The third Adult Treatment Panel of the National Cholesterol Education Program has recently issued revised guidelines for the treatment of cholesterol in adults. Increased attention to the metabolic syndrome and diabetes, including the inaccuracy of the low-density lipoprotein cholesterol (LDL-C) measurement in these patients because of elevated triglycerides is highlighted. To overcome the inaccuracy of the Friedewald equation in calculating LDL-C when the triglycerides are elevated, measuring non-high-density lipoprotein (non-HDL-C) may provide a better means to follow these patients toward their treatment goals. Recently, non HDL-C was shown to be a better predictor of cardiovascular death than LDL-C, even in patients with triglyceride levels below 200 mg/dL. The authors review the basis for using non-HDL-C as a treatment target for cholesterol, in comparison with other lipoproteins. PMID- 12091762 TI - Comparison of recovery patterns for patients undergoing coronary artery bypass grafting and minimally invasive direct coronary artery bypass in the early discharge period. AB - The purpose of this pilot study was to compare the postoperative problems, evaluation and response of symptoms, and functional status (physiologic and psychosocial functioning) during the early recovery period (2, 4, and 6 weeks after surgery) in 35 individuals who underwent coronary artery bypass grafting (n=24) or minimally invasive direct coronary artery bypass (n=11). The most frequent postoperative problem reported by the coronary artery bypass grafting group was an incisional infection (either sternal or leg); 26% reported infection at 2 and 4 weeks, and 21% at 6 weeks after surgery. Respiratory problems (pleural effusion, pneumonia) were the second most frequently reported problem, reported by 10% of the subjects at 2 and 4 weeks and by 16% at 6 weeks. Other, less frequent problems were severe nervousness, rhythm problems, and pericarditis. Minimally invasive direct coronary artery bypass patients reported fewer postoperative or cardiac-related problems, as only 5% indicated a problem with heart failure at both 2 and 4 weeks, and 36% reported being very nervous or having emotional problems at 4 weeks. Unlike postoperative problems, there were numerous similarities in postprocedural symptoms between these two groups. Fatigue, shortness of breath, and pain were the major symptoms reported postdischarge by both groups in this study. In addition, sleeping problems were also fairly prevalent in the coronary artery bypass grafting group, which is understandable, considering the fatigue ratings. Physiologic and psychosocial functioning varied minimally between the two procedures. While there were many similarities in the recovery patterns of both groups, the occurrence of postprocedural problems and symptoms of these two patient groups should be considered by clinicians to further tailor patient education. PMID- 12091761 TI - Factors influencing health-related quality of life in cardiac rehabilitation patients. AB - Associations of age, gender, cardiac procedure (coronary artery bypass grafting or percutaneous transluminal coronary angioplasty), risk stratification, and number of comorbidities with health-related quality of life (HRQL) were examined among 217 men and 84 women (mean age, 63+/-11 years) consecutively enrolled in a phase II cardiac rehabilitation program. Female gender and high-risk stratification were independently associated with impaired treatment gains in most areas of HRQL. Older age and having a greater number of comorbidities were also associated with impaired gains in specific areas. HRQL was generally lower at baseline but not post-treatment in coronary artery bypass grafting vs. percutaneous transluminal coronary angioplasty patients. Cardiac rehabilitation patients also had lower HRQL scores than an age-matched, normative sample at baseline (most areas) and post-treatment (mainly on role physical). Findings reinforce the need for cardiac rehabilitation programs to tailor assessments and risk factor modification strategies to patient characteristics associated with low levels of HRQL, especially in women and those with higher risk stratification. PMID- 12091763 TI - Therapeutic considerations in the management of patients with heparin-induced thrombocytopenia. PMID- 12091764 TI - Cardiovascular nursing research initiatives in Canada. PMID- 12091765 TI - Metabolic syndrome: a constellation of risk factors for vascular disease and diabetes with deadly consequences. PMID- 12091766 TI - When do we monitor for the effects of a "therapeutic" myocardial infarction? PMID- 12091767 TI - Clinical practice guidelines. PMID- 12091768 TI - Drug treatment of cardiovascular problems at elderly age. PMID- 12091769 TI - Pharmacologic therapy for the older patient with cardiovascular disease: new information and continued challenges. PMID- 12091770 TI - Issues of hormone replacement therapy and cardiovascular disease for elderly women. AB - Based on epidemiologic studies demonstrating mainly positive biologic effects of estrogen on cardiovascular risk factors and outcomes, earlier recommendations decreed that most, if not all women should be treated with long-term postmenopausal hormone replacement. A review of recent controlled clinical trials demonstrates that previously held dictums might not prove accurate. For elderly women, the decision to begin hormone replacement therapy should not be based on an assumption of protection from cardiovascular diseases. A careful assessment of the risks and benefits, as well as acknowledgement of the outcomes for which hormone replacement therapy has unknown impact, is needed for any decision to begin, continue, or stop hormone replacement therapy. PMID- 12091771 TI - High blood pressure in the geriatric population: treatment considerations. AB - Increases in blood pressure (BP), particularly systolic BP, have traditionally been considered to be a normal or "physiologic" component of the aging process. However, it is now clear that elevated BP, particularly systolic BP, represents a pathophysiologic manifestation of altered cardiovascular physiology and structure, ultimately manifesting as increased cardiovascular morbidity and mortality (myocardial infarction, stroke, and total cardiovascular death rates). More than one half of the population aged 65 or older have hypertension, defined as BP > or = 140/90 mm Hg. Framingham data indicate that the risk of coronary heart disease increases with lower diastolic BP at any level of systolic BP > or = 120 mm Hg, thus further stressing the importance of pressure-induced arterial vascular compliance changes and introducing pulse pressure as an important predictor of cardiovascular risk. Geriatric hypertension is generally of a salt sensitive nature and often associated with impaired baroreflex function. Reduction in sodium intake is important and effective in older patients, and should be initiated before or together with drug therapy. Encouraging data from clinical trials now strongly support the aggressive anti-hypertensive treatment of elderly patients. A recent meta-analysis of eight outcome trials evaluating the risks of treated and untreated isolated systolic hypertension has demonstrated a 30% reduction in combined fatal and nonfatal stroke, a 26% reduction in fatal and nonfatal cardiovascular events, and a 13% reduction in total mortality. Those drugs effective in younger patients also appear effective in the elderly; low-dose thiazides (alone or in combination with potassium sparing agents), beta blockers, long-acting dihydropyridine calcium antagonists, angiotensin-converting enzyme inhibitors, and angiotensin receptor blockers all have demonstrated efficacy. In selecting an agent, it is important to consider comorbid disease states, and to recognize the potential of all nonsteroidal anti inflammatory drugs, whether conventional or cyclooxygenase-2 specific, to increase BP or interfere with other antihypertensive agents. In general, the elderly should be treated to target BP levels identical to those suggested for younger patients, although a more gradual reduction to target, perhaps with an intermediate BP goal of < 160 mm Hg, may be advisable. PMID- 12091772 TI - Long-term pharmacologic management of atrial fibrillation in the elderly. AB - The first decision to be made in treating atrial fibrillation in the elderly is to determine whether to pursue a treatment strategy of rate control or rhythm control. Both strategies require use of anticoagulation therapy with warfarin (target international normalized ratio, 2.5; range 2-3). If a decision is made for rhythm control, the critical therapy is almost always with an antiarrhythmic drug. Before selecting an antiarrhythmic drug for use, it is first necessary to determine the presence or absence of underlying structural heart disease, as that will affect the available options for antiarrhythmic drug use. If there is no underlying structural heart disease, any of the available antiarrhythmic drugs may be used, although a clinically reasoned approach is suggested. If there is underlying structural heart disease, not all antiarrhythmic drugs are appropriate for use. A clinically reasoned approach is suggested in the presence of coronary artery disease, left ventricular dysfunction /congestive heart failure, or hypertension based largely on the risk/benefit profile of the drugs. PMID- 12091773 TI - Pharmacologic therapy of lipid disorders in the elderly. AB - Older men and women with coronary artery disease, prior stroke, peripheral arterial disease, and extracranial carotid arterial disease with a serum low density lipoprotein (LDL) cholesterol > 125 mg/dL despite diet should be treated with lipid-lowering drug therapy, preferably with statins, to reduce the serum LDL cholesterol to < 100 mg/dL. If statin drug therapy does not lower the serum LDL cholesterol to < 100 mg/dL in older persons with coronary artery disease, a bile acid binding resin, such as cholestyramine, should be added, since this drug does not increase the incidence of myositis in persons taking statins. The physician should use statins to treat older persons without atherosclerotic cardiovascular disease with a serum LDL cholesterol > or = 160 mg/dL plus one major risk factor, or a serum LDL cholesterol greater than or equal to 130 mg/dL plus a serum high-density lipoprotein (HDL) cholesterol < 50 mg/dL. Gemfibrozil may be useful in reducing the incidence of coronary events in persons with coronary artery disease whose primary lipid abnormality is a low serum HDL cholesterol level. There are no good data supporting treatment of hypertriglyceridemia unassociated with increased LDL cholesterol or decreased HDL cholesterol for prevention of cardiovascular disease. PMID- 12091774 TI - Peripheral arterial disease: epidemiology and drug therapy. AB - Lower-extremity peripheral arterial disease (PAD) is common among older men and women, affecting 20%-30% of older men and women in general medicine practices. History and physical examination are insensitive measures of PAD. However, PAD can be noninvasively and reliably diagnosed in the office with the ankle-brachial index, a ratio of Doppler-recorded systolic pressures in the lower and upper extremities. An ankle-brachial index less than 0.90, consistent with PAD, is associated with increased risk of cardiovascular morbidity and mortality in addition to functional impairment. Drug therapy in PAD is directed at reducing the increased risk of cardiovascular events and improving walking impairment. Intensive atherosclerotic risk factor intervention and angiotensin-converting enzyme inhibitors are recommended for reducing cardiovascular event rates in persons with PAD. Components of an effective exercise intervention and drug therapy to reduce claudication-related walking impairment in PAD are also reviewed. PMID- 12091775 TI - Ethical issues in the management of geriatric cardiac patients. PMID- 12091776 TI - Three kinds of block in a single ECG. PMID- 12091777 TI - Hour-glass left ventricle:Midventricular hypertrophy and apical aneurysm in elderly hypertrophic cardiomyopathy. PMID- 12091778 TI - [Psychiatric disorders in intensive care units]. AB - The diagnosis and treatment of psychiatric disorders in intensive care patients have been for a long time neglected. They are nowadays better recognized and managed. These disorders are mainly: delirium; anxiety disorders, from simple anxiety to panic disorder with agitation; adaptation disorders with depressive mood; brief psychotic disorders with persecution ideas. The manifestations of psychiatric disorders occur not only during the stay in intensive care unit (ICU) but also after transfer from ICU and several months after discharge from hospital. Part of psychiatric disorders is caused by organic or toxic causes (metabolic disturbances, electrolyte imbalance, withdrawal syndromes, infection, vascular disorders and head trauma). Nevertheless some authors estimate that they are due to the particular environment of ICU. The particularities of these units are: a high sound level (noise level average between 50 and 60 dBA), the absence of normal day-night cycle, a sleep deprivation, a sensory deprivation, the inability for intubated patients to talk, the pain provoked by some medical procedures, the possibility to witness other patients' death. Although most patients feel secure in ICU, some of them perceive ICU's environment as threatening. Simple environmental modifications could prevent the apparition of some psychiatric manifestations: efforts should be made to decrease noise generated by equipment and staff conversations, to provide external windows, visible clocks and calendar, to ensure adequate sleep with normal day-night cycle and to encourage more human contact. Psychotropic drugs are useful but a warm and empathetic attitude can be very helpful. Some authors described specific psychotherapeutic interventions in ICU (hypnosis, coping strategies.). To face anxiety, many patients have defense attitudes as psychological regression and denial. Patient's family is suffering too. Relative's hospitalization causes a crisis in family. Unpredicted illnesses often force family members to reorganize in order to regain their equilibrium. Every family should be proposed a psychological support. Caregivers can be distressed as well. This stress is due to their high responsibility and the fact that they face disease and death. Simple measures can lessen stress'effect and prevent the burn-out syndrome . In conclusion, the importance of a liaison psychiatrist-intensive care physician collaboration must be emphasized in order that patients and their family have a better psychological support. Psychological management should be proposed during the hospitalization and after discharge from hospital. PMID- 12091779 TI - [Chloral hydrate: a hypnotic best forgotten?]. AB - Synthesised by Justin Liebig in 1832 chloral hydrate is one of the oldest synthetic agents. Since 1869 it has been in use for hypnotic or sedative purposes. Chloral hydrate was used a lot from the end of the 19th century to the middle of the 20th century. Since then chloral hydrate has been less frequently in use as a hypnotic. In the 1990's, the principal use of chloral hydrate in pediatrics was the sedation of children for minor surgery during dental or diagnostic procedures. In general practice, it is an analgesia found in topical preparations. It was known as safe and easy to use. Now it is shown to be potentially dangerous (risk of death in case of intoxication) and there is doubt about genotoxicity and carcinogenecity. The pharmacological property was known in 1948 when Butler discovered the principal active metabolite, trichloroethanol. The gastro-intestinal tract rapidly absorbs chloral hydrate after oral or rectal use. The sedative and hypnotic effects appear in 20 to 60 minutes. The main metabolites [trichloroethanol (TCE) and trichloroacetic acid (TCA)] are formed by hepatocytes and erythrocytes. The half-life of chloral hydrate is short (a few minutes), the half lives of the metabolics are longer, 8 to 12 hours for TCE and 67 hours for TCA. The affinity for lipids is high. It is eliminated principally by the kidneys. Its mechanism of action is unknown. It is a depressor of the SNC, and the sedation is attributed to chloral hydrate and the hypnotic effect to TCE. The interactions appear with: alcohol, anticoagulants, amitriptyline and furosemide. The use of flumazenil (a gaba antagonist), in case of intoxication, indicates a possible action of GABA. The posology is usually between 0.5 to 2 g per day. Chloral hydrate is taken during meals to prevent gastric irritation. The main side effects are digestive, cardiologic (risk of rhythm disorder), dermatologic, neuropsychiatric (withdrawn, delusions, hallucination, dependence) and ophthalmologic. Death occurs after absorption of doses of around 10 g of hydrate chloral, some cases were reported with 5 g. The use of hydrate chloral is contra-indicated in cases of gastric ulcers, hepatic insufficiency, porphyry, respiratory insufficiency, association with anticoagulants and hyper sensibility. Nowadays should we be using chloral hydrate in cases of insomnia in adult and older people? A recent preclinical working group of the French Agency for evaluation of medicinal products reassessed the benefit/risk ratio of chloral hydrate. Many references are found about genotoxicity and carcinogenicity in recent literature. In France, since the end of 2000, chloral hydrate has been withdrawn from many medications for external use in dermatology and in stomatology. Chloral hydrate can be used as a pediatric sedative only once in a lifetime. The psychiatric indication for insomnia is no longer justified and especially in older people. PMID- 12091780 TI - [Anxiety and physical limitation: a complex relation]. AB - Psychological problems affecting physically handicapped individuals are understudied. However, some studies suggest a higher risk of developing an elevated level of anxiety among these individuals. A previous study reported that people with multiple sclerosis show more anxiety than patients suffering from another type of physical limitations. These results raise questions about the specificity of the link between anxiety and some medical conditions. The aim of the present study is to increase our understanding of manifestations of anxiety associated with degenerative illnesses. Three groups of patients with different physical limitations were compared on their level of anxiety and on various cognitive process. Groups were composed as: 1) 20 patients suffering from multiple sclerosis, 2) 18 patients diagnosed with another degenerative illness (either arthritis, muscular dystrophy or ataxia), and 3) 20 participants presenting a non-degenerative handicap. The 3 groups were not significantly different on age, sex and duration of the physical limitations. Symptoms of anxiety were measured with 3 questionnaires:1) the Hospital Anxiety and Depression Scale, 2) the Penn State Worry Questionnaire, used to assess the tendency to worry, and 3) the Worry Domains Questionnaire, assessing diversity and intensivity of worry themes. In turn, cognitive processes were evaluated with 4 questionnaires: 1) the Acceptance of Disability scale, used to assess the person's level of acceptance regarding a disability, 2) the Interpretation of Disturbing Thoughts questionnaire, an idiographical measure about the interpretation of thoughts associated with a physical handicap, 3) the Intolerance of Uncertainty scale, presenting beliefs about uncertainty and its consequences, and 4) the Cognitive Avoidance questionnaire, to evaluate the tendency of avoiding disturbing thoughts and images. Participants completed all questionnaires alone and the experimenter was available to answer any questions. MANOVAs were used to compare the 3 groups on the studied variables. Statistical analysis revealed no significant differences among groups for symptoms of anxiety and depression, tendency to worry and worry themes. Similarly, no significant differences were obtained on cognitive processes. The multiple sclerosis group and the other degenerative illness group were combined and compared to the non degenerative handicap group. MANOVAs conducted on symptoms and cognitive processes did not yield any significant differences. Results of the present study seem to indicate that the type of physical limitations is not an indicator of the presence of specific anxiety symptoms or cognitive processes. On an exploratory basis, all participants were compared according to their level of acceptance of disability. Results indicated that a lower level of acceptance was significantly associated with more anxious and depressive symptoms, excessive worries, and a greater intolerance of uncertainty. It seems that acceptance of disability plays a key role in maintaining psychological distress among people with a physical handicap. The influence of intolerance of uncertainty on acceptance of disability needs to be further explored. PMID- 12091781 TI - [The aging of Down's Syndrome subjects]. AB - The normal ageing of Down's Syndrome subjects is comparable to that observed in individuals who have an equivalent cognitive deficit. However it is earlier and is related to the former intellectual level and life story of the person. Before 50 years, there is no significant reduction of memory. After this age short-term memory, the speed of information processing and selective attention weaken. These changes are similar to those in older non-Down's Syndrome defective adults, giving the impression of early ageing in Down's Syndrome subjects. In terms of autonomy in everyday life, it is possible to establish an average evolutionary profile. From 60 years old, deterioration is estimated at 45% of the score obtained at 40 years, affecting in particular the skills necessary for the carrying out daily tasks (washing, dressing, feeding without assistance.). We have little knowledge of the psychiatric evolution of this people because older handicapped people are a new group in society. In the three fields of cognition, autonomy and mental health, the ageing of Down's Syndrome subjects is very sensitive to their environment. Pathological ageing of the Down's Syndrome subject is associated with the dementia syndrome that, with varying degrees, combines disorders of the cognitive functions and behavior, modifying the personality. The clinical diagnosis of dementia is difficult to establish in the Down's Syndrome subject and opinions diverge, also it is important to comply with three rules: 1) to establish an individual base line and to observe, with the help of regular evaluations, a clear change in performance; this must be confirmed by similar modifications in daily conducts; 2) the decline depends not only on the resources of the subject, but also on the demands made by environment; 3) lasting deterioration of capacities must be clearly greater than that observable in normal ageing to signify dementia. As a function of actual age, the Down's Syndrome shows early signs of ageing compared to the general population. One notes the presence of pathological anatomic lesions from 36 years old, which are similar to those observable among patients suffering Alzheimer Disease. However it seems that about 20% of Down's Syndrome subjects do not show clinical signs of dementia 20 years later. The diagnosis, which is delicate to establish, requires an ecological process consistent over time, underlining the influence of the context and the human environment on the ageing of the Down's Syndrome subject. PMID- 12091782 TI - [Methodology of a study on insomnia in the general population]. AB - The study of sleep disorders in the general population involves several methodological issues that need to be defined prior to proceeding to the epidemiological study. The rigor of the methodology is an important issue since it will determine the reliability of the data gathered. This paper describes the methodology used in an epidemiological study performed in the French general population using telephone interviews with the help of Sleep-EVAL, an expert system designed for this purpose. The study aimed to investigate the prevalence of insomnia disorders according to the DSM IV classification and the use of psychotropic medications in the general population. The methodological choices for this study were based on several considerations. First, the sample had to be representative of the French population. Second, the study had to be conducted in the shortest period of time. Third, the interviews had to be conducted with respect to a strict standardization and fourth, the realization costs had to be minimal for a maximum of data collected. The telephone interview procedure was chosen over postal and face-to-face interviews because it offered the possibility of conducting all the interviews from the same site. Supervision was easier. It also offered an absolute control in the application of the selection procedure. To draw the sample, a two-stage procedure was adopted. At the first stage, we pulled a random series of telephone numbers in each Nielsen region with respect to the size of the settlement. At the second stage, during the initial telephone contact, a household member was chosen using the Kish selection procedure. This method is based on the utilization of eight tables of selection that allows for the choice of the person to interview in a given household and keeps the representativeness of the sample. This technique is little used in telephone surveys because of its burden and its intrusive nature: the interviewer must collect the age and gender of all eligible subjects, to classify men from the oldest to youngest and then to classify women. However, it is the most rigorous selection method for epidemiological surveys. To reduce the refusal rates and to alleviate the work of interviewers, the Kish method was implanted in the computer software used for this study. For this study, the exclusion criteria were minimal. Only individuals younger than 15 years of age, those with a speech or hearing impairment and those who were too ill to perform the interview were not included. Subjects who refused to participate, those who hung up without speaking to the interviewer and those who hung up before completing at least half of the interview were tabulated as refusal. The participation rate was calculated by dividing the number of completed interviews by the number of eligible participants (completed interviews, refusals and telephone numbers where the interviewer was unable to determine if the individuals met an exclusion criterion). In this study, the participation rate was 80.8% (5 622 completed interviews/6 966 eligible households). The diagnostic tool used for this study was the Sleep-EVAL system, an expert system designed to conduct epidemiological studies in the general population. It is a level 2, non-monotonic system endowed with a causal reasoning able to provide sleep and mental disorders diagnoses according to the DSM IV classification for this study. Subsequent versions of Sleep-EVAL also included the International Classification of Sleep Disorders. System symbolic representation of the classifications was put in a compiled knowledge base. This knowledge base was read and interpreted by the inference engine at the beginning of the interview. During the interview, this interpretation changed as a function of the answers provided by the interviewee and by deductions made from the analysis of information the system already knew. All interviews began with a standard questionnaire about sociodemographic information and sleep habits. From these first answers, the Sleep-EVAL system emitted a series of diagnostic hypotheses that were confirmed or rejected with supplementary questions. The interview ended once all diagnostic possibilities were exhausted. The validity of the Sleep-EVAL system was demonstrated in different studies performed in sleep disorders clinics. There were several advantages in using such a tool to conduct epidemiological surveys. No special skills from the interviewers nor specific knowledge of sleep and mental disorders were required. All the questions were chosen and formulated by the Sleep-EVAL system. The interviewer had simply to read the questions as they appeared on the monitor screen and enter the interviewee's responses by clicking the appropriate answer or typing it on the keyboard. Missing answers were non-existent because there was no possibility of skipping a question or entering inconsistent answers. It also ensured the uniformity of the interviews. Furthermore, it allowed the exploration of infrequent diagnoses. In summary, the methodology used for this study allowed for the investigation of the sleep pathology of the French population in a short period of time: only three months were necessary to complete the 5 622 interviews. The use of a computerized tool greatly facilitated the training of the interviewers and also their work. Furthermore, it ensured a standardized administration of the interviews and the exploration of a broad range of disorders that could hardly be realized with traditional paper-pencil questionnaires. PMID- 12091783 TI - [May Medically Assisted Procreation (MAP) be relevant for homosexual women? Study among 147 gynaeco-logists involved in MAP techniques]. AB - The second part of the twentieth century has seen societal modifications as well as evolution of medical techniques allowing now thinking human procreation in terms of choices or even rights. Certain voices require sometimes Medically Assisted Procreation (MAP) for lesbians. Even though society did not allow such a possibility in France, it seemed interesting to question about it professionals actively involved in the use of MAP techniques. Through systematic internet queries, we obtained a list of one hundred private or public french medical institutions with a unit for the treatment of sterility. A telephone call to their secretary allowed us to individualize those doctors who did practice MAP. A sample of 147 medical doctors practicing MAP was then drawn. They were questioned with a clinical instrument including 20 ended-questions in order to assess their opinions on: homosexual women with a desire of a child; possibility for these clinicians to intervene with a donor insemination in such situations; developmental risk for such children. One hundred twenty five (85%) accepted to answer. Nine percent of these gynaecologists still consider homosexuality as pathological, and 10% as deviant - contrary to international classifications of mental disorders - and 5% deny good maternal abilities to homosexual women. Before the so-called french laws of bioethics in 1994, none of them had practiced a donor insemination for a lesbian couple, though 4% had realized some for single homosexual women. Two third of them do not agree opening donor insemination to homosexual women though for half of them, the anonymity of a donor is not perceived as prejudicial to the child. Eighty-seven percent of these gynaecologists think that a child brought up by homosexual parents is at risk for developmental disorder, the configuration supposed the most pathogenic being when the birth results from a donor insemination. The supposedly most important risk factors are thought to be the marginality of an homosexual family and the lack of a paternal figure at home. However, for 68% of the clinicians, this role can be taken by another male figure. These reasons make the gynaecologists reluctant to participate actively in the constitution of such a kind of family by the practice of a donor insemination. Even though demands of lesbian couples were not listed as an indication of donor insemination in the laws of bioethics, this does not seem to lessen the number of these demands in this population, and moreover if the law would allow this indication, half of these doctors would agree to practice it. The expression of the desire of a child by homosexual women and their request for its realization through medical techniques places the clinicians at the center of an ethical reflexion fed more by personal affects rather than scientific studies - however available - on the development of children brought up by an homosexual couple. Indeed, these studies indicate that these children suffer more from a societal view than parental sexual orientation, and it seems therefore appropriate to shed light on it in order to alleviate the weight of a stigmatization without any clinical argument founding it until now. PMID- 12091784 TI - [Vulnerability to depression in children and adolescents: update and perspectives]. AB - Depression in children and adolescents is associated with poor psychosocial functioning, high psychiatric comorbidity, risk of recurrent episodes or onset of bipolar disorder. These findings emphasize the importance of early identification of children and adolescents having elevated risk for future depression and further development, evaluation and greater availability of prevention strategies. Our review aims an update about depressive vulnerability in children and adolescents in the perspective of better identification of at-risk populations and targeting of prevention programs. Psychopathology, in particular anxiety and disruptive disorders are well identified risk-factors for later depression. Subclinical depressive symptomatology, also termed "demoralization", also identifies high-risk populations, likely to become incident cases of depression. It is still unclear whether this condition is prodromal depression, a specific clinical entity or the expression of biological and/or cognitive vulnerability. Familial risk for depressive disorders involves both genetic and psychosocial factors. Marital discord, poor communication and dysfunctional parenting practices are often present in families with affective disorders and can be implicated in increased depressive vulnerability in the offspring. Research on individual vulnerability in children and adolescents has focused on temperamental and cognitive characteristics. Temperament traits describe individual differences in reactivity and behavior. High emotionality, defined as the tendency to become upset easily and intensely has been associated with an increased risk for subsequent major depression. However, as the majority of high scorers will not become depressive cases, emotionality should not be the only criterion for selection of at-risk populations. Cognitive style including poor self esteem, low social competence and negative attributions are also associated with increased likelihood of depressive symptoms, but their predictive value for the onset of clinical depressive episodes needs further investigation. Familial and individual vulnerability is likely to heighten the depressogenic impact of life events and psycho-social adversity. Prevention interventions have been developed in the United States for children and adolescents at-risk for depression. In France, clinicians witness growing demands from families with affective illness concerned with risk of parent-child transmission of depressive vulnerability, prevention and early identification of symptoms. To meet this kind of emerging needs and to prevent family dysfunction, a preventive program targets offspring of depressed parents and uses clinician-based family approaches. Family and individual sessions aim a better understanding of illness experience and encourage the parents to identify and foster resilience in their children. Another type of preventive intervention focuses on children and adolescents with subclinical depressive symptoms, eventually associated with behavioral problems ou high level of parental conflict, recruited in school settings. These school based interventions combine cognitive and social problem-solving techniques. Both familial and school-based preventive interventions have proven applicable and promising in high-risk children and adolescents. Perspectives are more systematic identification of risk groups, including youngsters with past or current non affective symptoms who might benefit from depression prevention, long-term evaluation and cross-cultural applications of prevention programs. PMID- 12091785 TI - [Biological factors of PTSD: neurotransmitters and neuromodulators]. AB - Subjects with posttraumatic stress disorder exhibit abnormalities in many psychobiological systems. Studies of the effects of stress on learning and memory processes suggest that fear conditioning, behavioural sensitisation, and a failure of extinction may be important in the persistence and reexperiencing of traumatic memories and stress sensitivity. All these symptoms are supported by specific biological mechanisms including NMDA receptors, noradrenergic system, etc. Recent findings show that amino acid transmitters, glutamate and GABA, are intimately involved in the process of factual memory registration, and suggest that amine neurotransmitters, norepinephrine and serotonin, are involved in encoding emotional memory. In addition, research suggests that 5-HT neurones have directs effects on both adrenergic and HPA function. Such findings provide important information that should guide current pharmacological practice and the development of innovative biological therapeutics. PMID- 12091786 TI - [Review of psychopharmacological treatments in adolescents and adults with autistic disorders]. AB - Autism is an early developmental disorder. It leads to severe and durable disturbances. Given this problem, no treatment can be excluded a priori. Thus, many approaches are used to deal with autistic disorders. In France, pharmacological treatments are, for instance, largely and mostly used in adults. In the USA, these treatments concern 50% of persons with autism of any age. Nevertheless, they are rarely based on controlled studies. At the present, however, prescriptions and expected effects appear to be hard to localize. Furthermore, only few controlled studies validate their use. Aim - We offer a review of studies about medical treatments used in adolescents and adults with autism. They are classified in 3 categories: the first (category I) includes drugs used for their neurochemical effects focusing on autistic signs. The second (category II) covers drugs used for treatment of behavioural disorders frequently associated with autism. The third (category III) corresponds to a wide range of drugs or vitamins for wich only few case studies exist reporting irregular positive effects. The main hypothesis of this review is that autism involves a dysfunction of the neuromediation systems. This hypothesis opens new perspectives in the research of medical treatments in autism by focusing on molecules, which are supposed to have an effect on neuromediation systems. Method - Our review is based on studies, which have been published during the past twenty years. For many studies, data are limited to adolescents and adults. So we expanded our review to data available in children. The data bases that we have used are medline and psyclit. Keywords have been chosen according to: pharmacological considerations (psychotropic, psychoactive drugs, psychopharmacology) and clinical symptoms (autism, automutilations, aggressive behavior, and hyperactivity). Hypothesis of a dysfunction in the neuromediation systems in autism - Many studies exist about biochemical abnormalities in autism. As in schizophrenia and mental retardation, dysfunctions of the neuromediation systems are considered to be etiological factors. In 30% of people with autism the most regular dysfunction is the increase of serotonine. This led to the serotoninergic hypothesis in autism and to the use of active drugs in the serotonine system. However, the presence of other neurometabolic abnormalities also motivates the use of drugs, supposed to be active in other neuromediation systems. Pharmacological treatments in autism - Category I section sign 1 Active drugs in the dopamine system. Haloperidol (Dopamine antagonist): The effects of this molecule have been broadly studied in autism. Results indicate high efficiency in some symptoms of autism (lack in social behaviour, stereotypical behaviour) and in behavioural impairments that may be associated with autism (aggressive behaviour, hyperactivity). Its side effects, particulary the risk of late dyskinesy, make atypical antipsychotics preferable because of their lower risks. Risperidone (Dopamine and serotonine antagonist): Among several studies only few have been controlled. They indicate that Risperidone has positive effects on the behaviour and is quite well tolerated. section sign 2 Active drugs in the serotonine system. Clomipramine: after promising results, the medium-term efficiency has decreased and severe side effects have limited its use. Fluvoxamine, Fluoxetine, Sertraline (Specific serotonine drugs): Their efficiency has been mainly tested through open studies and their results are contrasted. In some cases, social behaviours have improved and aggressiveness and stereotyped behaviours have decreased. Fenfluramine: At present, this drug is removed from the market. Yet, some studies have suggested that it improves behavioural disturbances as well as performances in autism. section sign 3 Active drugs in the opiate system. Naltrexone: Several controlled studies have indicated an improvement in social and aggressive behaviours. Nevertheless, these studies have used small size sample and have not been replicated. Category II. This category correspond to drugs supposed to be active on neurochemical disturbances found in autism but their target symptoms are not autism specific signs as defined by the ICD 10. Buspirone: This serotonine agonist may have a good impact on emotional disorders and sleeping confusions. Methylphenidate: Most of the current studies about this noradrenergic drug concern children. The results are variable. Paradoxical effects may exist in children with severe mental retardation. Propanolol: Some isolated studies habe reported its efficiency on behavioural disturbances. Clonidine: This adrenergic drug treats efficiently some cases of aggressive behaviour and hyperactivity. Category III. This category contains a wide range of drugs, vitamins or method used in autism after sporadic observations of their positive effects. Secretine: An important improvement has been reported in isolated cases. However, controlled studies in children do not confirm these results. Vitamines B6, B12 and Magnesium: An improvement in socialization and in behavioural disorders have been reported in some cases, but these results are not yet confirmed. Lithium, Carbamazepine, Valproate: Results of some case studies have found it to be efficient in cyclic disorders. Gluten and casein free diet: An improvement of social behaviour have been reported by some parents after these diets. No controlled study has validated this observation. Conclusion - There is no consensus on the use of psychopharmacological treatments in autism. Although there exist many clinical observations, only few controlled studies have validated the efficiency and safety of these treatments. At the present time and until having sufficient studies, drugs are generally limited to severe disorders, for which usual psycho educational approaches are insufficient. PMID- 12091788 TI - [What is the interest of Klinefelter's syndrome for (child) psychiatrists?]. AB - Klinefelter's syndrome (KS) concerns men and is usually characterized by tallness, underdeveloped testes and sterility. It is generally due to the 47,XXY genotype, ie one extra X chromosome in each cell. Its estimated frequency among newborn boys is 1/500 to 1/700. It seems that 64% KS would be undiagnosed. Abnormally low levels of testosterone blood values are very common in this syndrome. In this case, replacement androgen therapy should be initiated (ideally at the age of 11-15) which prevents osteoporosis and enhances secondary sexual features. Case report - Since early childhood, Mr X has been shy, passive with few friends. When he was 13 years old, the school physician noted a delay of puberty and referred him to an endocrinologist who diagnosed KS. Androgen therapy was introduced but rapidly stopped, because the boy and his parents thought it was useless. Mr X consulted a psychiatrist at the age of 21. He presented a schizo-affective disorder with influence syndrome, auditory and visual hallucinations, labile mood with disinhibited and depressive periods. He was admitted in a psychiatry ward of a general hospital. An endocrinologist confirmed the diagnosis of KS and found very low blood testosterone levels. Besides lithium and risperidone which had already been introduced before the hospitalization, androgens (testosterone undecanoate) were very progressively given to Mr X with a daily psychiatric evaluation. One month after discharge, a major depressive episode led to the adjunction of citalopram. After one year of follow-up, Mr X shows increased social adjustment and enhanced interest; the influence syndrome has partially regressed and his mood is more stable. Discussion - In the years '60 and '70, systematic screenings in psychiatric hospitals have detected 1.3% KS among hospitalized boys, ie 10 times more than in the general population, and 0.6 to 1% KS among hospitalized men. A large variety of psychiatric disorders have been described. Boys presenting KS are usually described as shy, with little energy and initiative, and few friends. They cry more often than compares. Neuropsychological studies demonstrate significantly lower verbal IQ than controls, while performance IQ is generally normal and global IQ is in the normal range with large individual variations. Language acquisition is always delayed. However, agressiveness is not increased. In his follow-up study of 20 years, Nielsen at al found more psychiatric disorders among KS patients, compared to a group of hypogonadal patients at first examination (mean age=27 years). After 20 years follow-up, however, no significant difference remained between the two cohorts concerning the frequency of psychiatric hospitalizations or mental diseases. Several hypotheses have been proposed to explain psychological aspects of KS such as low levels of androgens during foetal and child development, personality disorder related to hypogonadism, delay of mitosis of cells with an extra X chromosome, but none of them is able to explain the specificity of psychological problems associated with KS. Concerning therapeutic aspects, specialists prone substitutive androgen therapy in case of too low testosterone blood levels, from the time of increase in FSH (around the age of 11-15). It prevents osteoporosis, backache and excessive tiredness often found in males with KS; testosterone also improves social drive, mood, concentration and ability at work. If KS diagnosis is made at adult age, androgen therapy has also shown some efficacy, though less than if started earlier. Due to the oral and written language problems of KS boys between 5 and 12 years of age, Graham et al. recommend anticipatory guidance for these boys. In addition, they insist on the importance of the information of the parents, language therapy, the reduction of the length of the instructions given by schoolmasters and specially stimulating and stable childhood conditions. Though it is generally thought that androgens increase agressiveness, we found no consistent data in litterature proving that the restoration of physiological androgen blood levels increases crimes nor aggressiveness. In the contrary, Miller and Sulkes described four cases of KS men presenting chronic fire-setting behaviors. Testosterone was introduced. For three of them, follow-up was available: their behavior seemed improved and none of them recurred. However, the initiation of androgen therapy for patients with severe psychiatric illness should be done very carefully. Conclusion - The Klinefelter's syndrome is frequent and, if not diagnosed (which seems to be the most common case), these men have higher risks to develop psychiatric disorders. Therefore, child psychiatrists and psychiatrists should evoke that diagnosis when they examine boys or men who present typical physical traits of KS (tallness, underdevelopped testes) associated to school problems and/or psychiatric disorders. Indeed, if the diagnosis is confirmed by an endocrinologist and a genetic testing, psychological follow-up and testosterone undecanoate treatment (in case of abnormal testosterone blood levels) should be initiated. This therapy generally improves physical well-being and improves mood, concentration, capacity at work. There is no consistent data in the litterature proving that restoring physiological testosterone blood levels would be dangerous for KS men presenting severe psychiatric troubles. However, this should be discussed in each situation with caution, and androgens should be introduced very progressively. PMID- 12091787 TI - [Treatment adherence in the recurrent depressive disorders]. AB - It is difficult to achieve treatment compliance in recurrent depressive disorders. This disease involves a combination of psychiatric and depressive disorders and is a chronic condition: all of the characteristic features of the disease constitute an obstacle to compliance. Compliance in depressive patients may be improved through various approaches. Information provided to patients, which is widely encouraged today both by doctors and by the regulatory authorities, must be as complete as possible; it must include discussion of factors of that favor depression, as well as the logical basis of the management approach proposed. Furthermore, the best guarantee of compliance is probably maintenance of euthymia by means of suitable treatment. Long-term therapy with antidepressants and the institution of a mood regulator (valproate in particular) have both been shown to be efficacious and significantly superior to placebo. Published studies show that continued treatment with an antidepressant can reduce the risk of depressive relapse noted during long-term follow-up (between 18 months and 5 years) by at least 50%. This has been demonstrated for both tricyclic antidepressants and specific serotonin reuptake inhibitors. The need for continued long-term administration of the dosages with proven efficacy during the acute episode has been demonstrated for imipramine but also appears to have been verified for SSRIs. Even when depressive relapse occurs, it is often less severe if the antidepressant treatment has been continued. Although first-line preventive therapy generally comprises long-term continued administration of the antidepressant, use of mood regulators may occasionally be proposed: valproic acid is generally preferred over lithium assaults on the ground of safety. However, there are few studies demonstrating the preventive efficacy of structured psychotherapy against recurrence of depression. Good treatment compliance entails major benefits: the prescribed treatment is able to reduce the risk of recurrence of depression, and in all probability also reduces subsequent vulnerability to depression, even after treatment discontinuation, as emphasized by the kindling theory of R. Post. PMID- 12091789 TI - [Compliance in schizophrenia: predictive factors, therapeutical considerations and research implications]. AB - Compliance has been defined as the extent to which a person's behavior coincides with the medical advice given. Medication compliance is one of the foremost problems affecting neuroleptic efficacy in psychiatric patients. Since chlorpromazine introduction in 1952, antipsychotics are the principal element of schizophrenia treatment. Actually progress links to the use of new antipsychotics are conditioned by quality of compliance. The problem of nonadherence to medication could concern 50% of prescription. The reported incidence of non compliance with antipsychotic medication ranges from 11 to 80%. In a two thirds of case rehospitalization is the result of complete or partial noncompliance. After one year of first hospitalisation, 40% of relapse results from non adherence to medication. Medication adherence problems increase hospitalisation, morbidity and mortality. Social consequences, professional problems and family troubles linked to hospitalisations lead to low quality of life for patients and high cost for society. There are three main methods of measuring compliance. These include patient and clinical self-report, pill counts, and biological measures. Self-report methods are generally the most cost-effective and time efficient way of obtaining an indication of compliance. In psychiatric research, the most commonly used self-report measure of compliance is the Drug Attitude Inventory (DAI) originally devised by Hogan et al. On the basis of criticism concerning DAI reliability, a new questionnaire of medication compliance was proposed: the Medication Adherence Rating scale (MARS). The main goal of compliance evaluation is to quantify this phenomenon with accuracy and to find predictive factors of medication nonadherence. Three types of factors influencing compliance are identified: factors due to medications, factors linked to patients and factors depending on the therapeutic relation with the clinician. Tolerance is considered as the principal reason explaining a bad compliance. Neurologic, endocrine and anticholinergic side-effects are the first fact of treatment stop. Medication prescription complexity is although important to take under consideration. Some psychotic's symptoms, comorbid addictive behavior, poor insight are mentioned in the case of noncompliance. Some effective actions to improve compliance are described. Information and communication with the patient, simplification of therapeutic plan, consultation planning and account of side effect are simple and effective actions. Social support is very important for improvement of compliance. The communication attitude of the clinician, therapeutic relation and prescription use are main points of compliance. Compared to a conventional care, psychoeducational programmes of compliance show their superiority. More research on compliance evaluation is needed. Information and tools must be proposed to practitioners. PMID- 12091790 TI - Hormones and homocysteine. AB - Hyperhomocysteinemia is a well-established risk factor for cardiovascular disease. Various factors, both modifiable and non-modifiable, interact with the homocysteine metabolism and determine the plasma homocysteine concentrations. These include genetic abnormalities, age, sex and various nutritional and hormonal determinants, all of which play a role in atherosclerosis and accelerated peripheral and cardio-vascular disease (CVD). Several hormones modulate homocysteine metabolism and hence may play a role in the pathogenesis of CVD. The mechanisms involved are unclear. The association of hyperhomocysteinemia with diabetes mellitus is complex and may explain some of the risk of CVD in diabetics not explained by traditional risk factors. Much conflicting data exists in the literature on the role of insulin on homocysteine metabolism, although insulin affects the enzymes regulating the homocysteine metabolism. Treatment of hyperhomocysteinemia with vitamins lowers plasma homocysteine concentrations. Little data is available on the effect of this intervention on cardiovascular outcomes. This review briefly outlines the homocysteine metabolism, summarizes its hormonal determinants, and discusses the role of hyperhomocysteinemia in diabetes, hyperlipidemia and other endocrine disorders. PMID- 12091791 TI - Kallikreins, steroid hormones and ovarian cancer: is there a link? AB - Kallikreins are a subgroup of serine proteases with diverse physiological functions. The human kallikrein gene family has now been fully characterized and includes 15 members tandemly located on chromosome 19q13.4. Strong experimental evidence supports a link between kallikreins and endocrine malignancies and especially, ovarian cancer. Three new kallikreins have been shown to be potential diagnostic and prognostic markers for ovarian cancer. Many other kallikreins are also differentially expressed in ovarian cancer, and preliminary reports underline their possible prognostic value. The mechanism by which kallikreins could be involved in ovarian cancer pathology is not known. A likely link could be their regulation through the steroid hormone receptor pathway. Most kallikreins are under sex steroid hormonal regulation in cancer cell lines. Given the co-expression of many kallikreins in ovarian cancer, it is reasonable to postulate that kallikreins are involved in a cascade enzymatic pathway that plays a role in cancer progression. A multiparametric kallikrein expression profile may be a useful tool for ovarian cancer diagnosis/prognosis when used either alone or in conjunction with existing markers. PMID- 12091792 TI - Advances in the treatment of diabetes-insulin analogues. AB - Insulin analogues are molecules derived by modifying the structure of the human insulin molecule, resulting in altered physico-chemical, biological and pharmacodynamic properties. The pharmacokinetic characteristics of the previously available rapid-, intermediate-, and long-acting preparations of human insulin make it almost impossible to achieve sustained normoglycemia. All currently available analogues have been shown to have a more physiological time-action profile with either a shorter onset and shorter duration of action (insulin lispro and insulin aspart) or a more constant effect lasting at least 24 hours (insulin glargine). These advantages in the time-action profiles have been shown to improve various surrogate parameters (e.g., postprandial blood glucose concentrations) in a number of randomized controlled trials. Insulin analogues also represent a unique tool to unravel structure-function relationships in insulin biochemistry and insulin action. Data on the currently available, currently tested and currently being developed analogs are reviewed. PMID- 12091794 TI - [Alteration of spermatogenesis and Y chromosome microdelations. Analysis of the DAZ gene family]. AB - The Y chromosome has a fundamental role in sex determination and regulation of spermatogenesis. Three regions (designated as AZFa, b, and c) on the long arm of this chromosome exist, deletions of which result in severe damage to spermatogenesis with azoospermia or severe oligozoospermia. Recent progresses in molecular biology and extraordinary development of assisted reproduction techniques contributed to the research on this chromosome and the genes involved in spermatogenesis. About 10-15% of subjects affected by azoospermia or severe oligozoospermia carry a deletion in one or more AZF regions, 60% of which involves AZFc. The genes responsible for the testicular phenotype observed in these subjects are DBY and USP9Y for AZFa, RBMY1 for AZFb, and DAZ for AZFc. In this article, the current knowledge on biology and genetics of the Y chromosome are reported with particular interest to deletions found in infertile subjects. Furthermore, the more recent advances on DAZ gene and its role in spermatogenesis and male infertility are discussed. PMID- 12091793 TI - [PAI-1, the primary plasmatic inhibitor of fibrinolysis. Physiopathologic role and molecular mechanisms]. AB - Plasminogen activator inhibitor 1 (PAI-1) is the primary physiological inhibitor of plasminogen activation in blood. PAI-1 is known to contribute to thrombus formation and to the development and the clinical course of acute and chronic cardiovascular diseases. Plasma levels of PAI-1 are regulated on a genetic basis but, more important, is the dependence on a series of other atherosclerotic risk factors like hypertriglyceridemia, diabetes and insulin resistance. The insulin resistance syndrome, which is characterized partly by obesity with visceral fat accumulation, is considered as a major regulator of PAI-1 expression. At least in vitro, insulin is a potent inducer of PAI-1 synthesis by human hepatic cells, and, we have recently disentangled the molecular mechanisms responsible for enhanced PAI-1 gene expression by insulin. However, clinical data fail to support a direct acute contribution of insulin in regulating circulating PAI-1 levels. Recently, it has been proposed that adipose tissue could be responsible for the elevated plasma PAI-1 level observed in insulin resistance. It now seems reasonable to view PAI-1 as one of the factors contributing to the complex gene environment interactions involved in the formation and dissolution of thrombi. PMID- 12091795 TI - [Obesity and low-grade systemic inflammation]. AB - The aim of this review is to deal with the significance of obesity as a promotor of a chronic low-grade inflammatory reaction favouring the development of atherosclerosis and cardiovascular diseases. Adipose tissue synthetizes and releases inflammatory cytokines involved in various atherothrombotic mechanisms and in glucose and lipid metabolism. A local renin-angiotensin system may partially support the obesity related hypertension. Most obese subjects had elevated plasma levels of inflammatory markers which correlate with the degree of obesity and insulin resistance and decrease after weight reduction and exercise. Some evidences suggest that long-chain polyunsaturated fatty acids and thiazolidinediones may be useful in preventing atherosclerosis. Obesity, by itself, has been considered an independent risk factor for cardiovascular diseases. The hypothesis that it is linked to the associated low-grade chronic inflammation is supported by the existence of altered indexes of chronic inflammation also in obese children who are free of other pathological conditions. Further research will be required to determine the pathophysiological meaning of the chronic inflammation associated to obesity. PMID- 12091796 TI - [Post-thyroidectomy hypocalcemia. Personal experience]. AB - BACKGROUND: The study evaluates the incidence of both early and protracted post thyroidectomy hypocalcemia over a 6-year period (1995-2000) in a series of 330 consecutive thryoid operations. The aim was to identify the profile of patients with the greatest risk of permanent parathyroid functional insufficiency and a suitable treatment protocol. METHODS: Having performed a statistical ana-lysis of all the variables, we divided the entire population of 330 patients into 4 groups based on the extent of demolition, the method of organotherapeutic replacement with L-Thyroxine and the (eco-radioguided) procedure used to identify the parathyroids: 59 patients undergoing subtotal thyroidectomy; 155 patients undergoing total thyroidectomy who commenced replacement organotherapy from postoperative day 15-30; 94 patients undergoing total thyroidectomy who commenced replacement organotherapy from postoperative day 1; 22 patients undergoing total thyroidectomy who underwent routine intraoperative parathyroid assay. RESULTS: The results show that the impact of more radical surgery in determining hypocalcemia, in particular protracted hypocalcemia, represents the most striking characteristic of patients with the greatest probability of permanent parathyroid insufficiency. We emphasise that early correction of postoperative hypothyroidism can drastically reduce the incidence of both early and protracted hypocalcemia. CONCLUSIONS: In conclusion, hypoparathyroidism is an exceptional event, whereas hypocalcemia is relatively common after total thyroidectomy. The effects of the early correction of postoperative hypothyroidism are undoubtedly beneficial, as is the use of eco- and radioguided methods for the intraoperative identification of the parathyroid glands, although they are not free from limits and criticism. PMID- 12091797 TI - One-day vs standard thyroidectomy. A perspective study of feasibility. AB - BACKGROUND: The aim of this randomized controlled trial is to evaluate the feasibility of one-day thyroidectomy, comparing the results of this method vs standard thyroidectomy. METHODS: From June 2000 to June 2001, 110 patients underwent total thyroidectomy under general anesthesia for thyroid disease. The patients were randomized into 2 groups: in group A (40 patients) we used the one day thyroidectomy; in group B (70 patients) we employed standard thyroidectomy. In both groups postoperative mobilization was immediate and the mean postoperative hospitalization stay was 21 hours (range: 18-24) in group A and 60 hours (range: 21-120) in group B. The mean follow-up was 10 months (range: 6-18 months). RESULTS: The patients of group A showed hypoparathyroidism with temporary hypocalcaemia in 3 cases (7.5%) vs 5 (7.1%) of group B; this finding was not statistically significant. No cases of definitive hypoparathyroidism, nor lesions of RLN, of the external branches of the superior laryngeal nerve, nor postoperative hemorrhage were observed in either group. CONCLUSIONS: The one-day thyroidectomy was found to be a safe, feasible and cost effective procedure, it is convenient for both the patient and the surgeon, and offers the same immediate and long-term results as the standard thyroidectomy in selected patients. PMID- 12091798 TI - The canine Phosducin gene: characterization of the exon-intron structure and exclusion as a candidate gene for generalized progressive retinal atrophy in 11 dog breeds. AB - PURPOSE: The exon-intron structure of the canine Phosducin (PDC) gene was identified and the gene evaluated as a candidate for generalized progressive retinal atrophy (gPRA) in 20 dog breeds. METHODS: Intronic sequences of the PDC gene were analyzed after amplification using polymerase chain reaction (PCR) and following sequencing from clones isolated from a canine genomic library. Mutation screening was performed by PCR with single strand conformation polymorphism (SSCP) analysis. Conspicuous banding patterns were analyzed via sequence analyses to detect the underlying nucleotide variations. RESULTS: No polymorphisms were identified after PCR-SSCP analysis within the entire coding region of the PDC gene. A 3 bp deletion in intron intervening sequence (IVS) 3 (position -16 to 18) was observed in 9 breeds, a T->A transversion (position IVS3 -63) in 10 breeds and an A->T transversion (position IVS3 -64) in 2 dog breeds. CONCLUSIONS PDC was excluded as a candidate gene for autosomal recessively (ar) transmitted gPRA in 11 of the 20 dog breeds investigated. PMID- 12091799 TI - Value, viability, and valves. PMID- 12091801 TI - Heroes and evidence. PMID- 12091802 TI - How do you study blue? PMID- 12091803 TI - The reoxygenation injury: is it clinically important? PMID- 12091804 TI - Cardiopulmonary bypass: Evidence or experience based? AB - OBJECTIVE: Evidence-based medicine is emerging as a new paradigm for medical practice. The purpose of this study was to evaluate the amount and quality of scientific evidence supporting principles that are currently applied for cardiopulmonary bypass performance. METHODS: A survey of all German departments of cardiac surgery regarding cardiopulmonary bypass performance disclosed major differences. Consequently, for 48 major principles of cardiopulmonary bypass performance, relevant Medical Subject Headings were identified, and a literature search of the Medline database was performed. Two sequentially applied sets of inclusion-exclusion criteria were selected to assess the best available evidence. RESULTS: Thirty-three thousand articles relating to the subject were identified. Among these, 1500 fulfilled the first set of inclusion criteria: meta-analysis of (randomized) controlled clinical trials and in vitro and animal studies. Rigorous methodological criteria were then applied to further select remaining publications. Ultimately, 225 articles referring to major cardiopulmonary bypass principles were identified as providing the best available evidence. These were graded according to their methodological rigor (susceptibility to bias). The scientific evidence on the investigated cardiopulmonary bypass principles did not prove to be of a high enough level to allow general recommendations to be made. CONCLUSIONS: The scientific data concerning the effectiveness and safety of key principles of cardiopulmonary bypass are insufficient in both amount and quality of scientific evidence to serve as a basis for practical, evidence-based guidelines. PMID- 12091805 TI - Long-term effects of surgical angiogenic therapy with fibroblast growth factor 2 protein. AB - OBJECTIVE: The long-term effects of surgical fibroblast growth factor 2 therapy are examined. METHODS: In a randomized, double-blind study, fibroblast growth factor 2 (10 microg or 100 microg) or placebo (n = 8 each) was delivered in the ungraftable myocardial territory of patients concomitantly undergoing coronary artery bypass grafting. Patients were followed up to 32.2 +/- 6.8 months postoperatively with clinical assessment and nuclear perfusion imaging. RESULTS: Baseline patient characteristics were similar between the 3 groups. There were 2 late deaths, one of pancreatic cancer and one of undetermined cause (both in the 100-microg fibroblast growth factor 2 group). Two patients (both in the control group) underwent a total of 6 repeat cardiac catheterizations for recurrent coronary events. Mean Canadian Cardiovascular Society angina class improved at late follow-up from baseline in all groups (P < or = .02); however, patients treated with either dose of fibroblast growth factor 2 had significantly more freedom from angina recurrence than those treated with placebo (P =.03). Late nuclear perfusion scans revealed a persistent reversible or a new, fixed perfusion defect in the ungraftable territory of 4 of 5 patients who received placebo versus only 1 of 9 patients treated with fibroblast growth factor 2 (P =.02). The overall sum of left ventricular stress perfusion defect scores was also lower in fibroblast growth factor 2-treated patients than in control subjects (1.3 +/- 1.4 vs 3.9 +/- 2.1, respectively; P =.04). A trend toward a higher late left ventricular ejection fraction was noted in fibroblast growth factor 2-treated patients (55.1% +/- 14.6% vs 44.3% +/- 6.5%, fibroblast growth factor 2-treated patients versus control subjects; P =.12). CONCLUSIONS: These data suggest that surgical angiogenic therapy with sustained-release fibroblast growth factor 2 may result in a prolonged myocardial revascularization effect that could translate into clinical benefit. PMID- 12091806 TI - Polymerized bovine hemoglobin solution as a replacement for allogeneic red blood cell transfusion after cardiac surgery: results of a randomized, double-blind trial. AB - BACKGROUND: Blood loss leading to reduced oxygen-carrying capacity is usually treated with red blood cell transfusions. This study examined the hypothesis that a hemoglobin-based oxygen-carrying solution can serve as an initial alternative to red blood cell transfusion. METHODS: In a randomized, double-blind efficacy trial of HBOC-201, a total of 98 patients undergoing cardiac surgery and requiring transfusion were randomly assigned to receive either red blood cell units or HBOC-201 (Hemopure; Biopure Corporation, Cambridge, Mass) for the first three postoperative transfusions. Patients were monitored before and after transfusion, at discharge, and at 3 to 4 weeks after the operation for subsequent red blood cell use, hemodynamics, and clinical laboratory parameters. RESULTS: The use of HBOC-201 eliminated the need for red blood cell transfusions in 34% of cases (95% confidence interval 21%-49%). Patients in the HBOC group received a mean of 1.72 subsequent units of red blood cells; those who received red blood cells only received a mean of 2.19 subsequent units (P =.05). Hematocrit values were transiently lower in the HBOC group but were similar in the two groups at discharge and follow-up. Oxygen extraction was greater in the HBOC group (P =.05). Mean increases in blood pressure were greater in the HBOC group, but not significantly so. CONCLUSION: HBOC-201 may be an initial alternative to red blood cell transfusions for patients with moderate anemia after cardiac surgery. In a third of cases, HBOC-201 eliminated the need for red blood cell transfusion, although substantial doses were needed to produce this modest degree of blood conservation. PMID- 12091807 TI - Alterations in left ventricular torsion in tachycardia-induced dilated cardiomyopathy. AB - OBJECTIVE: Left ventricular torsion reduces transmural systolic gradients of fiber strain, and torsional recoil in early diastole is thought to enhance left ventricular filling. Left ventricular remodeling in dilated cardiomyopathy may result in changes in torsion dynamics, but these effects are not yet characterized. Tachycardia-induced cardiomyopathy is accompanied by systolic and diastolic heart failure and left ventricular remodeling. We hypothesized that cardiomyopathy would alter systolic and diastolic left ventricular torsion mechanics, and this hypothesis was tested by studying sheep before and after the development of tachycardia-induced cardiomyopathy. METHODS: Implanted miniature radiopaque markers were used in 8 sheep to measure left ventricular geometry and function, maximal torsional deformation, and early diastolic recoil before and after rapid ventricular pacing was used to create tachycardia-induced cardiomyopathy. RESULTS: All animals had significant heart failure with ventricular dilatation and remodeling. With tachycardia-induced cardiomyopathy, maximum torsion relative to control conditions decreased (1.69 degrees +/- 0.61 degrees vs 4.25 degrees +/- 2.33 degrees ), and early diastolic recoil was completely abolished (0.53 degrees +/- 1.19 degrees vs -1.17 degrees +/- 0.94 degrees ). CONCLUSIONS: Cardiomyopathy is accompanied by decreased and delayed systolic left ventricular torsional deformation and loss of early diastolic recoil, which may contribute to left ventricular dysfunction by increasing systolic transmural strain gradients and impairing diastolic filling. Analysis of left ventricular torsion with radiofrequency-tagging magnetic resonance imaging should be explored to elucidate the role of torsion in patients with cardiomyopathy. PMID- 12091808 TI - Prevascularization with gelatin microspheres containing basic fibroblast growth factor enhances the benefits of cardiomyocyte transplantation. AB - OBJECTIVE: The effects of cell transplantation on the ischemic failing heart have already been documented. However, the area in and around infarct regions is not a good environment for cells to survive in because they are exposed to poor conditions in which certain requirements cannot be adequately supplied. We therefore designed a study to investigate the efficacy of prevascularization in ischemic regions before cell transplantation. METHODS: Rats with myocardial infarction were randomized into 4 groups: 11 rats received a culture medium injection to the left ventricular wall (control group), 11 received fetal cardiomyocyte transplantation (TX group), 11 received gelatin hydrogel microspheres incorporating basic fibroblast growth factor (FGF group), and 11 received basic fibroblast growth factor pretreatment sequentially, followed by cardiomyocyte transplantation (FGF-TX group). Four weeks later, left ventricular function was assessed by means of echocardiography and cardiac catheterization. RESULTS: In the FGF and FGF-TX groups neovascularization was found in the scar tissue 1 week later. The TX, FGF, and FGF-TX groups showed better fractional shortening than the control group (TX, FGF, FGF-TX, and control: 28% +/- 4.4%, 24% +/- 8.6%, 27% +/- 7.3%, and 17% +/- 4.6%, respectively; P <.01). Left ventricular maximum time-varying elastance was higher in the FGF-TX group than in the TX and FGF groups (FGF-TX, TX, and FGF: 0.52 +/- 0.23, 0.30 +/- 0.08, and 0.27 +/- 0.20 mm Hg/microL, respectively; P <.01). Histologically, more transplanted cells survived in the FGF-TX group than in the TX group. CONCLUSIONS: Prevascularization with basic fibroblast growth factor-incorporated microspheres enhances the benefits of cardiomyocyte transplantation. We expect that this system will contribute to regeneration medicine through its extensive application to other growth factors. PMID- 12091809 TI - Gadolinium attenuates regional stunning in the canine heart in vivo. AB - OBJECTIVE: Gadolinium, a lanthanide cation, ameliorates pathophysiologic features of both heart failure and cardiac arrhythmias. We have shown, in an in vitro model, that gadolinium blocks stretch-induced contractile dysfunction in both normal and stunned myocardium. The present study tested the hypothesis that gadolinium would also attenuate regional myocardial stunning in an in vivo model. METHODS: Mongrel dogs (n = 13) were subjected to regional myocardial ischemia (occlusion of the left anterior descending coronary artery) for 15 minutes, followed by reperfusion for 180 minutes. Intravenous gadolinium (500 micromol) was given to 7 dogs before ischemia; no gadolinium was given to control animals. Regional contractile function was assessed serially by means of both systolic shortening (percentage) and regional preload recruitable stroke work. RESULTS: Administration of gadolinium before ischemia had no effect on heart rate, arterial blood pressure, stroke volume, or regional contractile function. Ischemia resulted in paradoxical systolic bulging in both groups. After 180 minutes of reperfusion, systolic shortening was enhanced in gadolinium-treated animals compared with that in control animals (10.9% +/- 1.5% vs 2.4% +/- 1.7%, P =.003). Both the slope and x-axis intercept of regional preload recruitable stroke work returned to preischemic values in treated animals but remained abnormal in control animals. CONCLUSIONS: These data confirm that gadolinium attenuates regional myocardial stunning in vivo. Gadolinium may cause peripheral vasodilatation but does not appear to exert positive inotropic effects on the normal canine heart. The mechanism underlying gadolinium-mediated effects on stunned myocardium remains undefined, but this study suggests that use of gadolinium may represent a novel adjunct to current cardioprotective strategies. PMID- 12091810 TI - In vitro engineering of heart muscle: artificial myocardial tissue. AB - INTRODUCTION: Myocardial infarction followed by heart failure represents one of the major causes of morbidity and mortality, particularly in industrialized countries. Engineering and subsequent transplantation of contractile artificial myocardial tissue and, consequently, the replacement of ischemic and infarcted areas of the heart provides a potential therapeutic alternative to whole organ transplantation. METHODS: Artificial myocardial tissue samples were engineered by seeding neonatal rat cardiomyocytes with a commercially available 3-dimensional collagen matrix. The cellular engraftment within the artificial myocardial tissues was examined microscopically. Force development was analyzed in spontaneously beating artificial myocardial tissues, after stretching, and after pharmacologic stimulation. Moreover, electrocardiograms were recorded. RESULTS: Artificial myocardial tissues showed continuous, rhythmic, and synchronized contractions for up to 13 weeks. Embedded cardiomyocytes were distributed equally within the 3-dimensional matrix. Application of Ca(2+) and epinephrine, as well as electrical stimulation or stretching, resulted in enhanced force development. Electrocardiographic recording was possible on spontaneously beating artificial myocardial tissue samples and revealed physiologic patterns. CONCLUSIONS: Using a clinically well-established collagen matrix, contractile myocardial tissue can be engineered in vitro successfully. Mechanical and biologic properties of artificial myocardial tissue resemble native cardiac tissue. Use of artificial myocardial tissues might be a promising approach to reconstitute degenerated or failing cardiac tissue in many disease states and therefore provide a reasonable alternative to whole organ transplantation. PMID- 12091811 TI - Early and long-term results of the surgical treatment of tetralogy of Fallot with pulmonary atresia, with or without major aortopulmonary collateral arteries. AB - OBJECTIVE: We sought to determine the results of surgical treatment of patients with tetralogy of Fallot and pulmonary atresia with or without major aortopulmonary collateral arteries, to clarify variables affecting early and late mortality, and to expose late, nonfatal events affecting surgical patients. METHODS: The records of 495 patients operated on from 1977 to 1999 were reviewed. Patients were separated into those who did not undergo complete repair (group A) and those who did (group B). RESULTS: Group A consisted of 160 patients. Eighty one (51%) had palliative procedures, 45 (28%) had preliminary surgical stages (unifocalization and right ventricular outflow tract reconstruction) as initial operations, and 34 (21%) had all surgical stages but were rejected for complete repair. Early and late mortality were 16.3% (n = 26) and 23.1% (n = 31), respectively. Mean follow-up was 72.3 months. The presence of major aortopulmonary collateral arteries was a risk factor for late mortality (P =.0182). Group B consisted of 335 patients. Mean age at complete repair was 11.3 years (SD, 9.2). One hundred three (30%) patients had single-stage complete repair, whereas 232 (69%) had staged reconstruction. Twenty-two (6.6%) patients underwent reopening of the ventricular septal defect for high right ventricular pressure. Early and late mortality were 4.5% (n = 15). Risk factors were a peak right ventricular/left ventricular pressure ratio of greater than 0.7 and reopening of the ventricular septal defect (P < or = .05). Late mortality was 16% (n = 51). Mean follow-up was 11.4 years (SD, 7.5). Risk factors included male sex, nonconfluent central pulmonary arteries, reopening of the ventricular septal defect, and postrepair conduit exchange (n = 137). Ten- and 20-year results were an actuarial survival of 86% and 75% and freedom from reoperation of 55% and 29%, respectively. CONCLUSIONS: Surgical repair of patients with simple or complex forms of tetralogy of Fallot with pulmonary atresia can be achieved with low early mortality. Late mortality and need for reoperation, especially conduit replacement, continue to affect the long-term well-being of these patients. PMID- 12091812 TI - Percutaneous stent implantation to stenotic bioprosthetic valves in the pulmonary position. AB - BACKGROUND: We evaluated stent implantation across stenotic bioprosthetic pulmonary valves in 9 patients. METHODS: Nine patients (6 male patients) underwent stent implantation across stenotic bioprosthetic pulmonary valves between July 1996 and July 1999 at the Hospital for Sick Children, Toronto. Catheter intervention was indicated if echocardiography revealed Doppler estimates of right ventricular pressure of more than two thirds of systemic arterial pressure (or systolic septal flattening with an estimated gradient of >60 mm Hg across the valve prosthesis). Catheterization was performed during general anesthesia at an age (mean +/- SD) of 9.3 +/- 3.5 years and a weight of 32.0 +/- 17.1 kg 5.9 +/- 1.8 years after surgical insertion of a bioprosthetic valve in the pulmonary position: 7 patients with tetralogy of Fallot, 1 patient with congenital pulmonary stenosis-insufficiency, and 1 patient after a Rastelli operation. All had systolic septal flattening and right ventricular dilatation with moderate-to-severe pulmonary insufficiency before intervention. Fluoroscopy times were 33.1 +/- 9.5 minutes. Seven patients received a single P4014 stent, and 2 received single P308 stents (Palmaz; Johnson & Johnson Interventional Systems, Warren, NJ) without significant complications. RESULTS: The right ventricular systemic pressure decreased acutely from 83% +/- 16% to 41% +/- 10% (P <.001, n = 9), and the transvalvular gradient decreased from 49.7 +/- 8.5 to 11.0 +/- 5.9 mm Hg (P <.001, n = 8). During the follow-up period (10.9 +/- 8.1 months, n = 8), 1 patient had an unsuccessful attempt at redilation of the stent (right ventricular pressure, 60% systemic) and underwent uneventful surgical pulmonary valve replacement. None of the remaining patients had echocardiographic evidence of systolic septal flattening, and right ventricular dimensions did not change significantly. CONCLUSION: Stent implantation is a safe and effective means of providing palliative relief of obstructed bioprosthetic valves in the pulmonary position and can safely delay the requirement for pulmonary valve replacement. PMID- 12091813 TI - Homograft conduit failure in infants is not due to somatic outgrowth. AB - OBJECTIVE: It has been assumed that the need for homograft replacement is due to somatic outgrowth, but this has not been adequately studied. Our objective was to identify reasons for homograft conduit failure. METHODS: The records and imaging studies of 40 patients undergoing homograft conduit replacement of the right ventricular outflow tract from 1996 to 2000 were retrospectively reviewed. RESULTS: The majority of patients had a diagnosis of tetralogy of Fallot (n = 20) and truncus arteriosus (n = 13). The median age at the initial operation was 8 months (0.25-108 months). The initial homograft sizes ranged from 9 to 22 mm, and 28 conduits were of pulmonary origin. When comparing size of the initial homograft with patients' expected pulmonary valve diameter (z = 0), oversizing was noted to be +3 (range, 0.83-5.4). Median interval to conduit failure was 5.3 years (0.83-11.3 years). At homograft replacement, only 12 patients had an existing conduit that was 1 SD below the homograft conduit size needed (z < or = 1). Most conduits had important regurgitation, but this was rarely a primary reason for reintervention (n = 1). Reoperation was usually required for stenosis, with a median gradient of 53 mm Hg (20-140 mm Hg). Stenosis was further categorized angiographically as follows: homograft valvular stenosis (shrinkage; 21/40 [53%]), distal anastomotic stenosis (4/40 [10%]), conduit kinking (3/40 [8%]), sternal compression (3/40 [8%]), posterior shelf impingement (2/40 [5%]), and somatic outgrowth (3/40 [8%]). Replacement in 2 patients was for proximal hood aneurysm. Several patients (7/40 [18%]) had stenosis at multiple levels. The average decrease in conduit diameter was 47% (28%-73%). CONCLUSIONS: Somatic outgrowth is seldom a primary reason for homograft conduit replacement of the right ventricular outflow tract. The most common cause for failure is conduit obstruction with thickening and shrinkage at the annular area. Conduit stenosis was responsible for failure in 53% of patients, technical issues were responsible for 30%, and only 8% failed as a result of somatic outgrowth. Placement of a smaller homograft (z = 0) at the initial operation may decrease the incidence of conduit kinking, sternal compression, and posterior shelf impingement. PMID- 12091814 TI - Center-specific differences in mortality: preliminary analyses using the Risk Adjustment in Congenital Heart Surgery (RACHS-1) method. AB - OBJECTIVE: We sought to explore the usefulness of the Risk Adjustment in Congenital Heart Surgery method (designated RACHS-1) of adjusting for case-mix differences when comparing institutional mortality after surgery for congenital heart disease. METHODS: By using 1996 hospital discharge data from 6 states, centers performing at least 100 operations for congenital heart disease (patient age <18 years) were identified. Using the RACHS-1 method, procedures were grouped into 6 risk categories, and institutions were ranked in order of increasing mortality rate. A graphic display of ranks by risk category identified patterns of performance. Incorporating age, prematurity, and presence of a major noncardiac structural anomaly into multivariate models allowed computation of an overall risk-adjusted rank for each institution on the basis of its standardized mortality ratio. RESULTS: Among 109 centers performing 7177 operations for congenital heart disease, 22 performed at least 100 cases (72.3% of total operations). Unadjusted mortality rates ranged from 2.5% to 11.4%. A total of 4318 cases could be placed into 1 of the 6 risk categories. Few deaths occurred in risk category 1, and few institutions performed procedures in risk categories 5 and 6, making institutional comparisons in these categories uninformative. Considering mortality rates in categories 2 through 4, institutions displayed either relatively consistent ranks, a threshold increase in mortality as higher risk procedures were performed, or a threshold decrease in mortality. Standardized mortality ratios indicated which institutions performed better or worse than expected on the basis of their case mix. CONCLUSIONS: The RACHS-1 method can be used to judge relative institutional performance, either by evaluating within-risk-category differences or by comparisons of observed and expected mortality rates. PMID- 12091815 TI - Chronic hypoxia: a model for cyanotic congenital heart defects. AB - OBJECTIVE: The postoperative course of cyanotic children is generally more complicated than that of acyanotic children. A possible reason is reoxygenation injury at the beginning of cardiopulmonary bypass. In this study we tested the hypothesis that reoxygenation of chronically hypoxic hearts is worse than that of normoxic hearts. METHODS: Two groups of rats (n = 9 each) were exposed to either room air (fraction of inspired oxygen, 0.21%) or chronic hypoxia (fraction of inspired oxygen, 0.10%) for 2 weeks. Hearts were then isolated and perfused for 30 minutes with hypoxic buffer (oxygen saturation, 10%), followed by 30 minutes of reoxygenation (oxygen saturation, 100%). RESULTS: In hypoxic rats hematocrit values, hemoglobin concentrations, and red cells were higher (69% +/- 6% vs 40% +/- 6%, 219 +/- 14 vs 124 +/- 12 g/L, and 10.30 +/- 0.6 vs 6.32 +/- 0.5/microL/1000, respectively; P <.0001); the amount of ingested food was less (22.3 +/- 4.8 vs 30.7 +/- 3.9 g/d, P <.001), as was the amount of ingested water (21.0 +/- 3.1 vs 50.4 +/- 14.6 mL/d, P <.0001); and body weight was lower (182 +/ 14.2 vs 351 +/- 40.1 g, P <.0001), as was heart weight (1107 +/- 119 vs 1312 +/- 128 mg, P <.005). The heart weight/body weight ratio was higher (6.10 +/- 0.8 vs 3.74 +/- 0.1 mg/g, P <.0001). Systolic and diastolic functions, not different during the hypoxic baseline period, were more impaired in hypoxic than in normoxic hearts after the reoxygenation, whereas coronary resistance remained lower. During the hypoxic perfusion, the venous partial pressure of oxygen remained low in both groups, whereas during reoxygenation, partial pressure of oxygen was higher in hypoxic hearts, with a lower (P <.01) oxygen uptake. During hypoxic baseline adenosine triphosphate turnover, lactate production and lactate turnover were lower in hypoxic hearts (P <.005, P <.0001, and P <.0001, respectively). CONCLUSIONS: Body and blood values are severely affected by chronic hypoxia, and the cardiac effects of uncontrolled reoxygenation after chronic hypoxia are more severe than after acute hypoxia. PMID- 12091816 TI - Stage III non-small cell lung cancer and metachronous brain metastases. AB - OBJECTIVES: This study was undertaken to identify management strategies that maximize survival of patients with stage III non-small cell lung cancer and metachronous brain metastases and to determine whether any apparent improved survival was due to treatment or simply to patient selection. METHODS: Treatment evaluations of both primary non-small cell lung cancer and brain metastases were performed in 91 patients. Optimal treatment was identified by multivariable analysis. Propensity scoring and multivariable analysis were used to separate treatment benefit from patient selection. RESULTS: Risk-unadjusted median, 12-, and 24-month survivals were 5.2 months, 22%, and 10%, respectively. Younger age (P =.006), good performance status (P =.003), stage IIIA (P =.001), lung resection (P =.02), no other systemic metastases at time of diagnosis of brain metastases (P =.02), and either metastasectomy (P <.001) or stereotactic radiosurgery (P <.001) predicted best survival. However, metastasectomy or stereotactic radiosurgery was more common after lung resection (P =.02) and in patients with good performance status (P =.006), no other systemic metastases at time of diagnosis of brain metastases (P =.01), and fewer brain metastases (P <.001), suggesting that the patients with the best risk profile were selected for aggressive therapy of both lung primary and brain metastases. Despite this selection, analysis of propensity-matched patients demonstrated the benefit of lung resection and metastasectomy or stereotactic radiosurgery (P <.001). CONCLUSIONS: Younger patients with resected stage IIIA non-small cell lung cancer who have isolated metachronous brain metastases and good performance status do best when treated with metastasectomy or stereotactic radiosurgery. This survival benefit is a brain treatment effect, not the result of selecting the best patients for aggressive therapy. PMID- 12091817 TI - Replication-competent herpes virus NV1020 as direct treatment of pleural cancer in a rat model. AB - OBJECTIVE: Innovative treatments are needed for metastatic disease involving the pleura. NV1020 is a novel, multimutated, replication-restricted herpes simplex virus under investigation for its ability to selectively kill tumors by means of direct cell lysis. This study examines NV1020 in a rat model of pleura-based lung cancer. METHODS: Cytotoxicity and viral proliferation were evaluated in vitro by exposure of the human non-small cell lung cancer cell line A549 to virus. NV1020 was also tested in an in vivo pleura-based cancer model established by injecting 1 x 10(7) A549 cells into the thoracic cavity of nude rats. Intrapleural treatments (1 x 10(7) viral particles) were given 3 hours or 3 days after tumor injection to model treatment of microscopic or macroscopic disease (n = 8 9/group). Tumor burden was assessed at 5 weeks. NV1020 infection and dissemination within the thoracic cavity was determined by means of immunohistochemistry. RESULTS: In vitro, at multiplicities of infection (viral particles per tumor cell) of 0.01, 0.1, and 1.0, cell killing of A549 by NV1020 was 66%, 90%, and 97%, respectively, at 7 days after infection. Viral burst occurred by day 2. Intrapleural treatment was effective for both the microscopic (P <.001) and macroscopic (P <.05) in vivo tumor models. Virus was detectable by means of immunohistochemistry in tumors but not in adjacent normal intrathoracic tissues. CONCLUSIONS: NV1020 is not only highly cytotoxic to the human lung cancer line A549 in vitro but can be delivered in a clinically relevant fashion to safely and effectively treat pleura-based tumor in vivo in a rat model. PMID- 12091818 TI - Minimally invasive coronary artery bypass grafting versus percutaneous transluminal coronary angioplasty with stenting in isolated high-grade stenosis of the proximal left anterior descending coronary artery: six months' angiographic and clinical follow-up of a prospective randomized study. AB - OBJECTIVE: We sought to compare minimally invasive coronary artery bypass grafting (surgical intervention) with percutaneous transluminal coronary angioplasty with primary stenting (stenting) in patients having an isolated high grade stenosis (American College of Cardiology/American Heart Association classification type B2 or C) of the proximal left anterior descending coronary artery. At 6 months, both procedures were compared on the basis of quantitative angiography and clinical outcome. METHODS: Both treatments were compared in a single-center, prospective, randomized study. The primary end point of this study was quantitative angiographic outcome at 6 months. The secondary end point was 6 month clinical outcome. Statistical analysis was performed in accordance with the intention-to-treat principle. RESULTS: From March 1997 to September 1999, patients with angina pectoris caused by an isolated high-grade stenosis of the proximal left anterior descending coronary artery were randomly assigned to surgical intervention (n = 51) or stenting (n = 51). At 6 months, quantitative coronary angiography showed an anastomotic stenosis rate of 4% after surgical intervention and a restenosis rate of 29% after stenting (P <.001). Periprocedural events did not significantly differ between surgical intervention and stenting. After surgical intervention, 2 patients died; no patients died after stenting. After 6 months, no significant difference was found for major adverse cardiac or cerebral events and need for repeat target vessel revascularization. After 6 months, return of angina pectoris, physical work capacity, and use of antianginal drugs did not significantly differ between treatments. CONCLUSIONS: After 6 months, surgical intervention had a significantly better angiographic outcome than stenting in patients with an isolated high-grade stenosis of the proximal left anterior descending coronary artery. Clinical outcome did not significantly differ between treatments. PMID- 12091819 TI - The risk factors for deep and superficial chest surgical-site infections after coronary artery bypass graft surgery are different. AB - OBJECTIVE: We sought to determine risk factors for deep and superficial chest wound infections after coronary artery bypass graft surgery to develop predictive models. METHODS: We retrospectively analyzed data collected on 1980 consecutive patients undergoing coronary artery bypass surgery at our institution between January 1, 1996, and June 30, 1999, by using the Society of Thoracic Surgery database. Independent risk factors for surgical-site infection were identified with multivariate logistic regression. RESULTS: There were 37 (1.9%) deep chest and 46 (2.3%) superficial chest surgical-site infections. Obese diabetic patients had a 7.7-fold increased risk of deep chest infections after controlling for intra-aortic balloon pump use (odds ratio, 3.1) and postoperative transfusion (odds ratio, 2.3). Independent risk factors for superficial surgical-site infections included obesity (odds ratio, 3.1), diabetes in persons 65 years of age or older (odds ratio, 2.7), and current smoking (odds ratio, 2.5). Use of antiplatelet drugs was associated with a lower risk of superficial infections (odds ratio, 0.4). Predicted operative mortality as a marker of severity of illness was not clearly predictive of deep or superficial surgical-site infection. Mortality in the year after the operation was increased in patients with deep chest infections compared with that seen in uninfected control subjects (8/37 [21.6%] vs 114/1612 [7.1%], P =.004) but not in patients with superficial chest infections (7/47 [15.2%] vs 114/1612 [7.1%], P =.075). CONCLUSIONS: Risk factors for deep and superficial chest surgical-site infections after coronary artery bypass surgery differ, suggesting different mechanisms of pathogenesis. Appropriate risk stratification models specific to these important outcomes must be developed. PMID- 12091820 TI - Late hemodynamic and clinical outcomes of aortic valve replacement with the Carpentier-Edwards Perimount pericardial bioprosthesis. AB - OBJECTIVES: The aim of this study was to investigate the long-term clinical and hemodynamic outcomes after aortic valve replacement with the Carpentier-Edwards Perimount bioprosthesis (Edwards Lifesciences, Irvine, Calif), which has been used in our institution since 1984. METHODS: From January 1984 to December 1995, the Carpentier-Edwards pericardial bioprosthesis was used for aortic valve replacement in 254 patients (male/female ratio 117:137) with a mean age of 71 years (range 25-87 years). Before the operation, 216 patients (85%) were in New York Heart Association functional class III or IV. The predominant diagnosis was aortic stenosis (n = 219, 86%). Associated surgical procedures included coronary artery bypass grafting in 130 cases (51%), mitral valve replacement in 11 cases (4%), and tricuspid or mitral valve repair in 12 cases (5%). Previous cardiac operations had been performed in 36 cases (14%). Follow-up was 100% complete at a mean of 60 +/- 31 months. Univariate estimates of time-related cumulative probabilities were calculated by the Kaplan-Meier method. Multivariable adjustment was performed by Cox proportional hazards regression. Echocardiography was performed in 61% of long-term survivors. RESULTS: There were 11 early deaths (4%) and 58 late deaths. Actuarial survivals at 5, 10, and 12 years were 80% +/- 3%, 50% +/- 8%, and 36% +/- 9%, respectively. At 12 years the freedom from cardiac death was 73% +/- 7%, the freedom from valve-related death was 84% +/- 11%, the freedom from valve reoperation was 83% +/- 9%, the freedom from primary tissue failure was 86% +/- 9%, the freedom from thromboembolism was 67% +/- 13%, and the freedom from endocarditis was 98% +/- 1%. Echocardiography was performed on long-term survivors (mean follow-up 67 +/- 25 months) and showed that transvalvular peak and mean pressure differences measured with Doppler echocardiography were 23.2 +/- 9.6 and 12.3 +/- 4.8 mm Hg, respectively. Aortic regurgitation was found by Doppler echocardiography to be none or trivial, mild, moderate, and severe in 64%, 30%, 3%, and 1% of patients, respectively. Mean left ventricular mass index was 107.2 +/- 35.3 g/m(2) (118.9 +/- 40.2 g/m(2) in men and 98.8 +/- 28.8 g/m(2) in women) at late follow-up. One third of all patients, regardless of sex (n = 26/64 women and n = 14/45 men), had evidence of left ventricular hypertrophy. However, our analyses indicate that the residual left ventricular hypertrophy was not caused by valve mismatch but was probably multifactorial. CONCLUSION: The Carpentier-Edwards Perimount bioprosthesis has provided satisfactory clinical and hemodynamic outcome. However, at long-term follow-up about one third of the patients being investigated still had left ventricular hypertrophy examined by echocardiography. PMID- 12091821 TI - Aortic coarctation in the adult: management of complications and coexisting arterial abnormalities with hypothermic cardiopulmonary bypass and circulatory arrest. AB - OBJECTIVES: Adult patients with aortic coarctation may have complications after childhood repair. Other adult patients have coarctation and aneurysms of the aorta and the left subclavian artery. The optimal management of such cases is not clearly established. We evaluated the role of hypothermic cardiopulmonary bypass and circulatory arrest. METHODS: Thirteen adult patients (mean age 38 years) with coarctation and coexisting abnormalities of the aorta and left subclavian artery were treated. Five patients had pseudoaneurysms develop after bypass grafting (n = 3) or patch angioplasty (n = 2). These were detected a mean of 21 years (range 13-44 years) after the initial operation. Four pseudoaneurysms were asymptomatic, and 1 had ruptured. One patient had recurrent coarctation from fibrous obliteration of a 10-mm bypass graft inserted 15 years previously. The remaining 7 patients had aneurysms of the left subclavian artery (n = 5), aneurysms of the ascending aorta and arch (n = 1), or stenosis of the left subclavian artery (n = 1) in combination with moderate or severe coarctation. Resection and interposition graft replacement of the aneurysmal or stenotic aortic segments were performed in all cases with an interval of hypothermic circulatory arrest that averaged 44 +/- 5 minutes (range 33-54 minutes). Seven patients had interposition graft replacement of aneurysmal or stenotic left subclavian arteries. RESULTS: There were no in-hospital or late deaths (maximal follow-up 7 years). No patient had brain injury, paralysis, myocardial, respiratory, or renal failure. No patient has evidence of recurrent coarctation or aneurysm formation. CONCLUSIONS: Cardiopulmonary bypass with hypothermic circulatory arrest can safely be used in the treatment of complex adult coarctation. It permits accurate delineation of the anatomy with minimal dissection, avoidance of aortic clamping and sacrifice of intercostal arteries, precise interposition graft repair, and adequate protection of vital organs. PMID- 12091822 TI - Preoperative stress conditioning prevents paralysis after experimental aortic surgery: increased heat shock protein content is associated with ischemic tolerance of the spinal cord. AB - BACKGROUND: All forms of surgical therapy are stressful and injurious. The problems of paralysis, renal dysfunction, and colonic ischemia associated with aortic occlusion are due to acute ischemia-reperfusion injury at the cellular level. Acute-anterior spinal cord ischemia is the most devastating outcome of these iatrogenic-ischemic events. The majority of surgical procedures are performed electively and therefore provide an opportunity to preoperatively condition the patient to minimize these ischemia-related morbidities. OBJECTIVES: We sought to determine whether acute spinal cord injury associated with aortic occlusion can be prevented by induction of the cellular stress response by means of preoperative administration of whole-body hyperthermia or stannous chloride. METHODS: The study consisted of an experimental rabbit model of infrarenal aortic occlusion for 20 minutes at normothermic body temperature. RESULTS: Control rabbits experienced an 88% (7/8) incidence of paralysis after spinal cord ischemia induced by 20 minutes of aortic occlusion, whereas animals treated preoperatively with either whole-body hyperthermia (0/9) or stannous chloride (0/4) never became paralyzed (P <.001 for control vs treated groups). Ischemic protection of the spinal cord was associated with increased content of stress proteins within tissues of pretreated animals. CONCLUSION: Prior induction of the heat shock response in the whole animal will increase the content of stress proteins within the spinal cord and other tissues and result in the prevention of hind-limb paralysis associated with aortic occlusion. We have designated the preoperative induction of the cellular stress response for the prevention of ischemic tissue injury stress conditioning. We suggest that stress-conditioning protocols represent the opportunity to practice preventative medicine at the molecular level. PMID- 12091824 TI - An arterial switch operation for a concordant crisscross heart with the complete transposition of the great arteries. PMID- 12091823 TI - Clinical-pathologic conference in general thoracic surgery: bilateral lung transplantation for sarcoidosis with aspergilloma. PMID- 12091825 TI - Severe aortic valve fusion after nearly three years of support with the Novacor left ventricular assist system. PMID- 12091826 TI - Endovascular treatment of ruptured thoracic aortic aneurysms. PMID- 12091827 TI - Acute chest pain 16 days after coronary artery bypass surgery: an unusual cause. PMID- 12091828 TI - Repair of coexistent infrarenal and thoracoabdominal aortic aneurysm: combined endovascular and open surgical procedure with visceral vessel relocation. PMID- 12091829 TI - The Achilles heel of composite arterial grafting: early occlusion of the distal right coronary limb. PMID- 12091830 TI - Minimally invasive explantation of implantable left ventricular assist devices. PMID- 12091831 TI - Undifferentiated sarcoma of the heart: a rare clinicopathologic presentation. PMID- 12091832 TI - Totally autologous Ross procedure. PMID- 12091833 TI - Primary myxofibrosarcoma of the esophagus. PMID- 12091834 TI - Mediastinal chyloma after esophageal cancer resection: an unusual complication causing left cardiac failure. PMID- 12091835 TI - Extensive epidural cufflike growth of malignant pleural mesothelioma causing spinal cord compression. PMID- 12091836 TI - Right postpneumonectomy syndrome and severe pectus excavatum in a child: surgical management. PMID- 12091837 TI - Intussusception and spontaneous amputation of the esophagus. PMID- 12091838 TI - Postoperative synthetic human atrial natriuretic peptide infusion and oral spironolactone administration for a patient with giant atria and low plasma level of atrial natriuretic peptide. PMID- 12091839 TI - Optimizing neurologic outcome in coronary bypass surgery. PMID- 12091840 TI - Impact of aerosolized prostacyclin analog for severe pulmonary hypertension in thoracic organ transplantation. PMID- 12091841 TI - Promoting clerkship directors: the importance of evidence. PMID- 12091842 TI - Erythropoietin treatment in extremely low birth weight infants: blood in versus blood out. PMID- 12091843 TI - Is failure to thrive a greater concern than obesity? PMID- 12091844 TI - Early treatment with erythropoietin beta ameliorates anemia and reduces transfusion requirements in infants with birth weights below 1000 g. AB - OBJECTIVE: To investigate whether recombinant erythropoietin (rhEPO) reduces the need for transfusion in extremely low birth weight (ELBW) infants (birth weight 500-999 g) and to determine the optimal time for treatment. METHODS: In a blinded multicenter trial, 219 ELBW infants were randomized on day 3 to one of 3 groups: early rhEPO group (rhEPO from the first week for 9 weeks, n = 74), late rhEPO group (rhEPO from the fourth week for 6 weeks, n = 74), or control group (no rhEPO, n = 71). All infants received enteral iron (3-9 mg/kg/day) from the first week. The rhEPO beta dose was 750 IU/kg/week. Success was defined as no transfusion and hematocrit levels never below 30%. RESULTS: Success rate was 13% in the early rhEPO group, 11% in the late rhEPO group, and 4% in the control group (P =.026 for early rhEPO versus control group). Median transfusion volume was 0.4 versus 0.5 versus 0.7 mL/kg/day (P =.02) and median donor exposure was 1.0 versus 1.0 versus 2.0 (P =.05) in the early rhEPO group, the late rhEPO group, and the control group, respectively. Infection risk was not increased and weight gain was not delayed with rhEPO beta. CONCLUSION: Early rhEPO beta treatment effectively reduces the need for transfusion in ELBW infants. PMID- 12091845 TI - Efficacy of albuterol inhalation in treatment of hyperkalemia in premature neonates. AB - OBJECTIVE: To evaluate the efficacy of inhaled albuterol for treatment of hyperkalemia in premature neonates by conducting a prospective, randomized placebo-controlled and double-blinded clinical trial. STUDY DESIGN: Neonates <2000 g receiving mechanical ventilation with central serum potassium > or =6.0 mmol/L (6.0 mEq/L), were randomly assigned to treatment or placebo groups. Albuterol (400 microg) or saline was given by nebulization. The dose was repeated every 2 hours until the potassium level fell below 5 mmol/L (maximum 12 doses) or there were signs of toxicity. RESULTS: Nineteen patients completed the study (8 in the albuterol and 11 in the saline group). Serum potassium levels declined rapidly in the first 4 hours in the albuterol group, from 7.06 +/- 0.23 mmol/L to 6.34 +/- 0.24 mmol/L (P =.003) versus no significant change in the saline group (6.88 +/- 0.18 mmol/L to 6.85 +/- 0.24 mmol/L; P =.87). At 8 hours, the fall continued to be greater in the albuterol group versus the saline group (5.93 +/- 0.3 mmol/L and 6.35 +/- 0.22 mmol/L, respectively; P =.04). CONCLUSION: Albuterol inhalation may be useful in rapidly lowering serum potassium levels in premature neonates. PMID- 12091846 TI - Results of systematic voiding cystourethrography in infants with antenatally diagnosed renal pelvis dilation. AB - OBJECTIVES: To assess the yield of neonatal renal ultrasonography as a predictor of abnormalities on voiding cystourethrography (VCUG). STUDY DESIGN: We prospectively followed 264 infants with antenatal renal pelvis dilation. Two successive neonatal renal ultrasound examinations were performed at day 5 and 1 month after birth. VCUG was performed in all infants. RESULTS: Neonatal ultrasound findings were abnormal in 190 infants (72% of total). Among them, 63 (33%) had an abnormal VCUG. The most common abnormalities were primary vesico ureteral reflux (VUR) in 30 infants, VUR into the lower pole of a duplex kidney in 14, refluxing megaureter in 9, posterior urethral valves in 5, and ureterocele in 5. When both neonatal ultrasound findings were normal (74 infants), the VCUG showed abnormalities in only 5 (6.7%) patients (4 low-grade primary VUR and 1 posterior urethral valve). The sensitivity, specificity, positive predictive value, and negative predictive value of 2 successive ultrasound scans in the neonatal period to predict an abnormality on VCUG were 93%, 35%, 33%, and 93%, respectively. CONCLUSIONS: A normal-appearing urinary tract on 2 successive neonatal ultrasound scans rarely coexists with abnormal findings at VCUG. Therefore, in such patients, VCUG does not seem justified. PMID- 12091847 TI - Influence of congenital heart disease on survival in children with congenital diaphragmatic hernia. AB - OBJECTIVE: We sought to assess outcome in patients with CDH and HD to determine if LHR is also predictive of outcome in this subset of patients. STUDY DESIGN: We carried out a retrospective review (April 1996-October 2000) of patients with isolated CDH (n = 143, 82.2%) and patients with HD (n = 31, 17.8%) to determine the incidence of additional anomalies, survival to term, CDH repair, cardiac repair, and survival to discharge. Survival based on LHR was analyzed in a subset of fetuses. RESULTS: The risk of death from birth to last follow-up was 2.9 times higher for patients with CDH plus HD than for patients with CDH alone (P <.0001). Of 11 patients with CDH plus HD who had CDH repair (5 of whom also had HD repair), 5 survived. All 10 patients with an LHR <1.2 died; 3 of 6 with an LHR >1.2 survived (Fisher exact test, P =.04). CONCLUSION: Heart disease remains a significant risk factor for death in infants with CDH. The LHR helps predict survival in this high-risk group of patients. PMID- 12091848 TI - Efficacy, tolerability, and kinetics of lamotrigine in infants. AB - OBJECTIVES: To investigate the efficacy, tolerability, and kinetics of lamotrigine during the first year of life. STUDY DESIGN: We studied 13 infants with intractable seizures; 7 had partial seizures and 7 had infantile spasms (1 had both). Patients received open-label lamotrigine as add-on therapy for 3 months. Seizure frequency, response ratio, and side effects score were determined and compared with the baseline period. RESULTS: The rate of partial seizures per day decreased from 8.57 +/- 2.29 to 4.00 +/- 2.15 (P =.027) and infantile spasms from 8.71 +/- 2.15 to 3.61 +/- 2.762 (P =.028). Apparent clearance increased during the first year of life, with a break point at 2 months of age (mean, 0.119 +/- 0.021, 0.217 +/- 0.094 L/h per kilogram for infants <2 months and those 2 to 12 months old, respectively,P <.001). Twenty-four-hour concentration to time plots of three 3- to 4-week-old neonates showed a half-life of 23.44 +/- 3.57 hours. Compared with a group of 17 older children, LTG had similar efficacy (response ratios, -0.68 +/- 0.12 and -0.74 +/- 0.11, P =.504), and similar adverse effects scores (0.67 +/- 0.67 and 0.23 +/- 0.166, P =.95). CONCLUSIONS: Lamotrigine is a useful and well tolerated drug for partial seizures and infantile spasms in infants <1 year of age. However, lamotrigine has age dependent kinetics that must be taken into consideration. PMID- 12091850 TI - Prediction of outcome after hypoxic-ischemic encephalopathy: a prospective clinical and electrophysiologic study. AB - OBJECTIVES: To evaluate and compare the predictive value of history, clinical examination, and biologic and electrophysiologic data regarding the prognosis of children with acute hypoxic-ischemic encephalopathy (HIE). STUDY DESIGN: Prospective cohort of 57 consecutive children who were mechanically ventilated for HIE throughout a 3-year period in a tertiary pediatric intensive care unit at a university hospital in France. RESULTS: At 24 hours after admission, 12 patients had died, 3 were awake and 42 showed impaired consciousness or were in a coma, of whom 38% had a favorable outcome. In this group, an initial cardiopulmonary resuscitation duration longer than 10 minutes and a Glasgow Coma Scale <5 at 24 hours after admission were associated with an unfavorable outcome (positive predictive value [PPV] 91%, 100%; sensitivity 50%, 54%). A discontinuous electroencephalogram (EEG), the presence of spikes or epileptiform discharges were associated with an unfavorable outcome (PPV 100% for the 2 criteria; sensitivity 27%, 54%). The bilateral absence of the N20 wave on short latency sensory evoked potentials (SEPs) had a PPV for unfavorable outcome of 100% (sensitivity 63%). CONCLUSIONS: The clinical assessment combined with EEG and SEPs allow an early prediction of the prognosis of children with HIE. PMID- 12091849 TI - Effect of combination antiretroviral therapy on cerebrospinal fluid HIV RNA, HIV resistance, and clinical manifestations of encephalopathy. AB - OBJECTIVES: This study evaluated the effect of treatment with abacavir/lamivudine/zidovudine versus lamivudine/zidovudine on cerebrospinal fluid (CSF) human immunodeficiency virus (HIV) RNA and clinical manifestations of HIV encephalopathy in children. STUDY DESIGN: HIV-infected children 7 months to 10 years of age (n = 23) were studied. CSF and plasma were obtained at baseline and weeks 8, 16, and 48. Genotype analysis of HIV was attempted at baseline and week 48. Neurologic evaluations were performed at baseline and weeks 16, 32, and 48. RESULTS: At baseline, 83% of children had >2.00 log(10) copies/mL HIV RNA in CSF, but only 10% had HIV RNA measurable at week 48. Among children in whom paired genotyping of HIV was possible, 8 of 11 had identical patterns in both CSF and plasma at baseline, whereas at week 48, only 1 of 9 children had similar patterns. Neurologic abnormalities were observed in 83% of children at baseline but only 35% of children at week 48 (P =.004), suggesting a benefit of treatment. CONCLUSIONS: Antiretroviral therapy was associated with a decline in CSF HIV RNA and an improvement in neurologic status. The development of genotypic mutations was different in CSF and plasma, suggesting discordant viral evolution. These results suggest that antiretroviral treatment in children should include agents with activity in the CNS. PMID- 12091851 TI - Predictors of developmental disabilities after open heart surgery in young children with congenital heart defects. AB - OBJECTIVES: To determine the prevalence of persistent developmental impairments in children with congenital heart defects and to identify factors that enhance risk for an adverse outcome. STUDY DESIGN: Eligible infants (n = 131) <2 years of age requiring open heart surgery were recruited prospectively. Subjects were assessed during surgery and again 12 to 18 months later with standardized developmental assessments and formal neurologic examinations. RESULTS: Mean age at follow-up testing was 19.1 +/- 6.6 months. Assessments indicated that 41% had abnormal neurologic examinations. Gross and/or fine motor delays were documented in 42%, and 23% demonstrated global developmental delay. Univariate and multiple regression models identified the following factors increasing the risk for persistent developmental deficits: preoperative and acute postoperative neurodevelopmental status and microcephaly, type of heart lesion, length of deep hypothermic circulatory arrest, age at surgery, and days in the intensive care unit (P <.05). CONCLUSIONS: Children with congenital heart defects commonly have ongoing neurologic, motor, and developmental deficits well after surgical correction. The cause is multifactorial and includes brain injury before, during, and after heart surgery. PMID- 12091852 TI - QTc interval prolongation and QTc dispersion in children and adolescents with type 1 diabetes. AB - OBJECTIVES: To evaluate whether QT interval, QT interval corrected for heart rate (QTc), and QTc dispersion changes are already present in children and adolescents with diabetes. STUDY DESIGN: QT interval, QTc, and QTc dispersion were measured on a 12-lead surface electrocardiogram in 60 children and adolescents with stable type 1 diabetes and in 63 sex- and age-matched control subjects. Differences were evaluated by using the Kolmogorov-Smirnov Z test. The number of patients with QTc > 440 ms was compared in the two groups. The possible influence of age, sex, diabetes duration, and glycosylated hemoglobin (HbA(1c)) was examined by using Spearman correlation analysis. RESULTS: Diabetic children had significantly longer QTc intervals and a significantly larger QTc dispersion. The number of individuals with a QTc >440 ms was significantly higher in the diabetic group (14/60) than in the control group (2/63). The effect of age on R-R interval and QTc dispersion in healthy children was less pronounced in children with diabetes. HbA(1C) values did not significantly correlate with any of the parameters. CONCLUSIONS: QTc prolongation and a larger QTc dispersion are already present in a significant proportion of children and adolescents with diabetes. PMID- 12091853 TI - Physiologic regulators of bone turnover in young women with anorexia nervosa. AB - OBJECTIVE: To clarify the role of physiologic regulators of bone turnover in patients with anorexia nervosa (AN). STUDY DESIGN: Adolescent girls with AN (n = 61) had anthropometric, nutrition, and exercise data acquired, and bone mineral density (BMD) and body composition measured by dual energy x-ray absorptiometry. Serum samples were obtained for hormones, proresorptive cytokines, and bone formation markers, and urine for bone resorption markers. RESULTS: In bivariate correlation analyses, significant (P <.05) predictors of lumbar BMD included height, weight, and exercise. In multiple regression models, these significant relationships held, even after controlling for the duration of amenorrhea and AN. For total body BMD, the same positive predictors were found and percentage of body fat was a negative correlate. For hip BMD, exercise and weight were found to be positive predictors. Dehydroepiandrosterone sulfate (DHEAS) was inversely correlated with N-telopeptides (NTx), and insulin-like growth factor I (IGF-I) was directly correlated with osteocalcin. Proresorptive cytokine levels were low or undetectable. CONCLUSIONS: Exercise and weight were positive predictors of BMD. These data are the first to suggest a relationship between DHEAS and increased bone resorption in AN. IGF-I was correlated with bone formation indices. Low cytokine levels suggest that these factors do not mediate the increased bone resorption of AN. PMID- 12091854 TI - Prevalence of methylphenidate use and change over a two-year period: a nationwide study of 2- to 11-year-old Canadian children. AB - OBJECTIVE: To provide age- and sex-specific estimates of methylphenidate use and to determine use changes over a 2-year period. STUDY DESIGN: We examined the first and second data collection cycles of the National Longitudinal Survey of Children and Youth, a Canadian household survey of children. Participants were children aged 2 years to 11 years at both the first and second cycles whose mothers responded; thus, 16,798 (13,059) children were assessed for the first (second) cycle. Logit modeling was used to estimate prevalence of methylphenidate use, to determine sex and age effects on prevalence, and to examine use changes from cycle 1 to 2. RESULTS: Methylphenidate prevalence ranged from 0.09% to 3.89% across 2- to 11-year old children from the first cycle. Boys were 4.6 times more likely than girls to consume methylphenidate. Use was >4 times greater among 6- to 7-year-old children compared with 4- to 5-year-old children and almost 2 times greater among 8- to 9-year-old children compared with 6- to 7-year-old children. Methylphenidate prevalence increased by 36% from cycle 1 to 2. CONCLUSIONS: Methylphenidate prevalence was relatively low. Boys and school-age children had higher rates of methylphenidate use, and use among 2- to 11-year-old children appeared to be increasing over time. PMID- 12091855 TI - Recurrent pain, emotional distress, and health service use in childhood. AB - OBJECTIVES: To determine the psychosocial correlates of recurrent pediatric pain and its relationship to health service use and medical presentations for "unexplained" symptoms in primary care. STUDY DESIGN: Children 4 to 15 years of age who complained frequently of aches and pains to parents were compared with those with infrequent or no pain on measures of demographics, psychopathology, school attendance and performance, perceived health, and service use. Univariate analysis was followed by logistic regression. RESULTS: Children who complained often of aches and pains used more health services, had more psychosocial problems, missed more school, and did worse academically. After controlling for health service use and demographics, recurrent pain was significantly associated with negative parental perceptions of child health and the presence of internalizing psychiatric symptoms. Higher levels of ambulatory health service use were associated with negative perceptions of child health, recurrent pain, visits for "unexplained" symptoms, and internalizing psychiatric symptoms. CONCLUSIONS: Pediatric recurrent pain challenges traditional service delivery models characterized by segregated systems of care for physical and mental disorders. Longitudinal and psychobiological studies of the relationship between recurrent pain, internalizing psychopathology, and health beliefs are warranted to direct future treatment efforts. PMID- 12091856 TI - How useful is fecal pancreatic elastase 1 as a marker of exocrine pancreatic disease? AB - OBJECTIVES: To evaluate the role of fecal elastase 1 (E1) as a marker of exocrine pancreatic insufficiency (PI). STUDY DESIGN: Fecal E1 was measured in patient groups with (1) failure to thrive but no pancreatic or intestinal disease (disease control patients); (2) PI; (3) pancreatic sufficiency; and (4) steatorrhea caused by a variety of intestinal diseases. RESULTS: Fecal E1 in all disease control patients exceeded 200 microg/g stool. Only 1 (2%) of 50 patients with PI exceeded the minimum reference value of 100 microg/g stool. In contrast, 3 (11%) of 28 patients with pancreatic sufficiency (with Shwachman-Diamond syndrome) had fecal E1 concentrations <100 microg/g stool, as did 5 (20%) of 25 patients with steatorrhea from intestinal causes, all of whom had diluted feces caused by short gut. CONCLUSIONS: Fecal E1 is a useful noninvasive screening test of PI in childhood. A negative test (>100 microg/g stool) had 99% predictive value for ruling out PI. However, a positive test in those with short gut or Shwachman-Diamond syndrome must be interpreted with caution. PMID- 12091857 TI - Premature pubarche in girls is associated with functional adrenal but not ovarian hyperandrogenism. AB - OBJECTIVE: We hypothesized that there would be evidence of functional ovarian hyperandrogenism in girls with premature pubarche (PP) at diagnosis. METHODS: White girls <8 years of age and black girls <6 years with PP (n = 15) were studied. Prepubertal girls (n = 13; 5.3-10.9 years) and early pubertal girls (n = 8) served as control subjects. The biochemical marker for functional ovarian hyperandrogenism was the 17-hydroxyprogesterone (17-OHP), androstenedione (AD), and estradiol (E2) response to subcutaneous leuprolide during adrenal suppression with dexamethasone. This was studied in girls with PP and in control subjects. RESULTS: ACTH stimulated 17-hydroxypregnenolone (17-OH Preg), dehydroepiandrosterone (DHEA), and AD levels, and 17-OH Preg:17-OHP and DHEA:AD ratios were significantly higher in girls with PP than in prepubertal control subjects (n = 18) (P < or =.003). The ovarian response to leuprolide stimulation was comparable in girls with PP and prepubertal control subjects, but the response in prepubertal study subjects was significantly lower than in pubertal control subjects (P =.016 for Delta17-OHP, P =.001 for DeltaAD, and P =.026 for DeltaE2). CONCLUSIONS: Contrary to the hypothesis, PP in girls was not associated with prepubertal evidence of ovarian hyperandrogenism but was associated with functional adrenal hyperandrogenism. PMID- 12091858 TI - Time course to hypothyroidism after fixed-dose radioablation therapy of Graves' disease in children. AB - OBJECTIVE: To characterize the development of hypothyroidism in pediatric patients who receive a fixed dose of radioactive iodine (RAI). STUDY DESIGN: Medical records of children treated with fixed-dose RAI for Graves'disease between 1993 and 2001 were reviewed. Multiple variables including sex, age, thyroid hormone levels, thyroid-stimulating immunoglobulin titer, antithyroid medication use, and 24-hour radioiodine uptake were investigated as possible predictive factors for the development of hypothyroidism after treatment. All patients received RAI at a dose of between 13.8 and 15.6 mCi (average, 14.7 mCi; SD, 0.5). RESULTS: Permanent hypothyroidism developed in all 40 patients, although a second dose of RAI was required in one case. The average time to hypothyroidism was 77 days (SD, 32), with a range of 28 to 194 days; 75% of the patients were diagnosed with hypothyroidism between 40 and 90 days. RAI treatment was ineffective in an additional patient, who required subtotal thyroidectomy. CONCLUSIONS: We conclude that a fixed dose of RAI is effective therapy in nearly all pediatric patients with Graves'disease. Factors predicting the time course to hypothyroidism were not identified. PMID- 12091859 TI - Low-density areas on high-resolution computed tomograms in chronic pediatric asthma. AB - OBJECTIVE: In children with chronic persistent asthma, we evaluated whether the presence of increased residual volume (RV) after anti-inflammatory treatment correlates with the detection of low-density areas on high-resolution computed tomography (HRCT), similar to those in emphysema. METHODS: Children with a confirmed diagnosis of asthma (n = 32) were enrolled in a prospective study. All patients had reduction of airflow in the peripheral airways, increased RV, and increased serum eosinophil cationic protein (ECP) values indicating airway inflammation. All the children were treated with salmeterol (50 microg twice daily) and fluticasone (250 microg twice daily) for a 3-month period. RESULTS: At the end of treatment, peripheral eosinophil counts, serum ECP, forced expiratory volume in 1 second (FEV(1)), mean forced expiratory flow during the middle half of forced vital capacity (FEF(25-75)), RV, and total lung capacity values improved in all the patients. HRCT was normal in 22 children (68.8%); in the remaining 10 subjects, low-density areas were found despite normalization of FEV(1), FEF(25-75), and significant reduction in ECP. A significant correlation was found between persistence of RV values >150% predicted and the presence of low-density areas on HRCT (r = 0.84, P <.0001). CONCLUSIONS: Structural changes similar to emphysema are also present in asthmatic children. Our findings suggest that the persistence of increased RV may be used to identify subjects with low density areas on HRCT. PMID- 12091861 TI - Pneumomediastinum and subcutaneous emphysema in an 18-month-old child. PMID- 12091860 TI - Prevalence and correlates of household exposures to tobacco smoke and pets in children with asthma. AB - OBJECTIVES: To examine the prevalence rates of household smoking and ownership of a furred or feathered pet, the intercorrelation of these home environment measures, and their association with sociodemographic, family, and child asthma variables. STUDY DESIGN: Children with asthma (n = 152, aged 7-18 years) with asthma and their primary parent were evaluated through the use of reliable and valid questionnaires focusing on exposure to household smoke and furred or feathered pets as well as sociodemographic, family, and asthma variables. RESULTS: Prevalence of household smoking and pet ownership were high and comparable to normal levels in the US population. Smoking and pet ownership were not correlated with each other or with asthma medication adherence. Sociodemographic, family, and asthma variables showed distinct patterns of correlation with smoking and pet status. Household smoking was associated with poorer family resources and greater stress; pet ownership was associated with greater resources. CONCLUSIONS: Smoke exposure and pet ownership are not related to one another in children with asthma and will require independent counseling strategies because they relate in different and opposite ways to socioeconomic status. PMID- 12091862 TI - Disease severity at time of referral for pediatric failure to thrive and obesity: time for a paradigm shift? AB - The percent of ideal body weight at first visit to pediatric referral clinics was compared by chart review between patients with failure to thrive and obese patients. Results indicated that underweight children were mildly underweight, whereas the overweight children were severely so. In primary care settings, underweight children may be referred more aggressively than overweight children. PMID- 12091863 TI - Long-term prevention of premature coronary atherosclerosis in homozygous familial hypercholesterolemia. AB - Homozygous familial hypercholesterolemia (FHH) leads to severe premature atherosclerosis. A 22-year-old woman with FHH has been treated with a combination of H.E.L.P. apheresis (heparin-mediated extracorporeal LDL precipitation) and statins for 15 years. The combined treatment maintained a plasma LDL-cholesterol reduction from baseline of 840 to 122 mg/dL (85% reduction). In addition, H.E.L.P. apheresis reduced the elevated lipoprotein(a) and fibrinogen levels by 60% to 70%. All xanthomata disappeared. There is no evidence of premature atherosclerosis studied by means of electron beam computed tomography and (13)N ammonia positron emission tomography: The entire coronary vasculature is free of calcifications. Her myocardial blood flow at rest (87 mL/100 g/min) and during stress (308 mL/100 g/min) and the coronary flow reserve (3.5) are normal after H.E.L.P. treatment. This case demonstrates the efficacy and safety of the combined treatment of H.E.L.P. apheresis and statins even in serious cases of FHH. PMID- 12091864 TI - Persistent hyperlipasemia caused by macrolipase in an adolescent. AB - Serum amylase and lipase frequently rise during bouts of acute pancreatitis, and measurement of these enzymes provides important diagnostic information. We report a pediatric patient with persistent elevations of serum lipase resulting from macrolipasemia, a complex of lipase with IgG, rather than pancreatitis. PMID- 12091865 TI - Airway nitric oxide in Duchenne muscular dystrophy. AB - The expression of muscle membrane-associated neuronal nitric oxide synthase (NOS1) is significantly impaired in Duchenne muscular dystrophy. Mean (+/- SEM) exhaled NO in 13 male patients with Duchenne muscular dystrophy was significantly lower than in 11 healthy age-matched male control subjects (7.5 +/- 1.4 vs 16.6 +/- 3.2 parts per billion, P <.02) or 17 adult male control subjects (18.5 +/- 1.8 parts per billion, P <.001). These findings provide indirect evidence that NOS1 contributes significantly to fractional exhaled nitric oxide in healthy children. PMID- 12091866 TI - Physical therapy and cognitive-behavioral treatment for complex regional pain syndromes. AB - Complex regional pain syndromes (CRPS; type 1, reflex sympathetic dystrophy, and type 2, causalgia) involve persistent pain, allodynia, and vasomotor signs. We conducted a prospective, randomized, single-blind trial of physical therapy (PT) and cognitive-behavioral treatment for children and adolescents with CRPS. Children 8 to 17 years of age (n = 28) were randomly assigned to either group A (PT once per week for 6 weeks) or group B (PT 3 times per week for 6 weeks). Both groups received 6 sessions of cognitive-behavioral treatment. Assessments of pain and function were repeated at two follow-up time periods. Outcomes were compared at the three time points through the use of parametric or nonparametric analysis of variance and post hoc tests. All five measures of pain and function improved significantly in both groups after treatment, with sustained benefit evident in the majority of patients at long-term follow-up. Recurrent episodes were reported in 50% of patients, and 10 patients eventually received sympathetic blockade. Most children with CRPS showed reduced pain and improved function with a noninvasive rehabilitative treatment approach. Long-term functional outcomes were also very good. PMID- 12091867 TI - Thrombus in the left ventricle of a newborn. PMID- 12091868 TI - Nailfold capillaroscopy in Henoch-Schonlein purpura: a follow-up study of 31 cases. PMID- 12091869 TI - VCUG timing. PMID- 12091870 TI - How much iron to supplement and when. PMID- 12091871 TI - Effectiveness of growth hormone treatment in short children with chronic renal failure. PMID- 12091872 TI - Neonatal lung injury and "new BPD": need for a new terminology. PMID- 12091873 TI - A novel fucose recognition fold involved in innate immunity. AB - Anguilla anguilla agglutinin (AAA), a fucolectin found in the serum of European eel, participates in the recognition of bacterial liposaccharides by the animal innate immunity system. Because AAA specifically recognizes fucosylated terminals of H and Lewis (a) blood groups, it has been used extensively as a reagent in blood typing and histochemistry. AAA contains a newly discovered carbohydrate recognition domain present in proteins of organisms ranging from bacteria to vertebrates. The crystal structure of the complex of AAA with alpha-L-fucose characterizes the novel fold of this entire lectin family, identifying the residues that provide the structural determinants of oligosaccharide specificity. Modification of these residues explains how the different isoforms in serum can provide a diverse pathogen-specific recognition. PMID- 12091874 TI - Structure of the immunodominant surface antigen from the Toxoplasma gondii SRS superfamily. AB - Toxoplasma gondii is a persistent protozoan parasite capable of infecting almost any warm-blooded vertebrate. The surface of Toxoplasma is coated with a family of developmentally regulated glycosylphosphatidylinositol (GPI)-linked proteins (SRSs), of which SAG1 is the prototypic member. SRS proteins mediate attachment to host cells and interface with the host immune response to regulate the virulence of the parasite. The 1.7 A structure of the immunodominant SAG1 antigen reveals a homodimeric configuration in which the dimeric interface is mediated by an extended beta-sheet that forms a deep groove lined with positively charged amino acids. This basic groove seems to be conserved among SRS proteins and potentially serves as a sulfated proteoglycan-binding site on target cell surfaces, thus rationalizing the promiscuous attachment properties of Toxoplasma to a broad range of host cell types. PMID- 12091875 TI - Early organization of pre-mRNA during spliceosome assembly. AB - Intron excision from precursor mRNAs (pre-mRNAs) in eukaryotes requires juxtaposition of reactive functionalities within the substrate at the heart of the spliceosome where the two chemical steps of splicing occur. Although a series of interactions between pre-mRNAs, pre-spliceosomal and spliceosomal factors is well established, the molecular mechanisms of splicing machinery assembly, as well as the temporal basis for organization of the substrate for splicing, remain poorly understood. Here we have used a directed hydroxyl radical probe tethered to pre-mRNA substrates to map the structure of the pre-mRNA substrate during the spliceosome assembly process. These studies indicate an early organization and proximation of conserved pre-mRNA sequences during spliceosome assembly/recruitment and suggest a mechanism for the formation of the final active site of the mature spliceosome. PMID- 12091876 TI - Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism of immune evasion. AB - B7-H1, a recently described member of the B7 family of costimulatory molecules, is thought to be involved in the regulation of cellular and humoral immune responses through the PD-1 receptor on activated T and B cells. We report here that, except for cells of the macrophage lineage, normal human tissues do not express B7-H1. In contrast, B7-H1 is abundant in human carcinomas of lung, ovary and colon and in melanomas. The pro-inflammatory cytokine interferon-gamma upregulates B7-H1 on the surface of tumor cell lines. Cancer cell-associated B7 H1 increases apoptosis of antigen-specific human T-cell clones in vitro, and the apoptotic effect of B7-H1 is mediated largely by one or more receptors other than PD-1. In addition, expression of B7-H1 on mouse P815 tumor increases apoptosis of activated tumor-reactive T cells and promotes the growth of highly immunogenic B7 1(+) tumors in vivo. These findings have implications for the design of T cell based cancer immunotherapy. PMID- 12091877 TI - Revascularization of ischemic tissues by PlGF treatment, and inhibition of tumor angiogenesis, arthritis and atherosclerosis by anti-Flt1. AB - The therapeutic potential of placental growth factor (PlGF) and its receptor Flt1 in angiogenesis is poorly understood. Here, we report that PlGF stimulated angiogenesis and collateral growth in ischemic heart and limb with at least a comparable efficiency to vascular endothelial growth factor (VEGF). An antibody against Flt1 suppressed neovascularization in tumors and ischemic retina, and angiogenesis and inflammatory joint destruction in autoimmune arthritis. Anti Flt1 also reduced atherosclerotic plaque growth and vulnerability, but the atheroprotective effect was not attributable to reduced plaque neovascularization. Inhibition of VEGF receptor Flk1 did not affect arthritis or atherosclerosis, indicating that inhibition of Flk1-driven angiogenesis alone was not sufficient to halt disease progression. The anti-inflammatory effects of anti Flt1 were attributable to reduced mobilization of bone marrow-derived myeloid progenitors into the peripheral blood; impaired infiltration of Flt1-expressing leukocytes in inflamed tissues; and defective activation of myeloid cells. Thus, PlGF and Flt1 constitute potential candidates for therapeutic modulation of angiogenesis and inflammation. PMID- 12091878 TI - Hyporesponsiveness to vaccination with Borrelia burgdorferi OspA in humans and in TLR1- and TLR2-deficient mice. AB - The Lyme disease vaccine is based on the outer-surface lipoprotein (OspA) of the pathogen Borrelia burgdorferi, and 95% of vaccine recipients develop substantial titers of antibodies against OspA. Here, we identified seven individuals with very low antibody titers after vaccination (low responders). The macrophages of low responders produced less tumor necrosis factor-alpha and interleukin-6 after OspA stimulation and had lower cell-surface expression of Toll-like receptor (TLR) 1 as compared to normal cells, but normal expression of TLR2. TLRs activate innate responses to pathogens, and TLR2 recognizes lipoproteins and peptidoglycan (PGN). After OspA immunization, mice genetically deficient in either TLR2 (TLR2( /-)) or TLR1 (TLR1(-/-)) produced low titers of antibodies against OspA. Notably, macrophages from TLR2(-/-) mice were unresponsive to OspA and PGN, whereas those from TLR1(-/-) mice responded normally to PGN but not to OspA. These data indicate that TLR1 and TLR2 are required for lipoprotein recognition and that defects in the TLR1/2 signaling pathway may account for human hyporesponsiveness to OspA vaccination. PMID- 12091879 TI - Direct effects of interleukin-13 on epithelial cells cause airway hyperreactivity and mucus overproduction in asthma. AB - Asthma is an increasingly common disease that remains poorly understood and difficult to manage. This disease is characterized by airway hyperreactivity (AHR, defined by exaggerated airflow obstruction in response to bronchoconstrictors), mucus overproduction and chronic eosinophilic inflammation. AHR and mucus overproduction are consistently linked to asthma symptoms and morbidity. Asthma is mediated by Th2 lymphocytes, which produce a limited repertoire of cytokines, including interleukin-4 (IL-4), IL-5, IL-9 and IL-13. Although each of these cytokines has been implicated in asthma, IL-13 is now thought to be especially critical. In animal models of allergic asthma, blockade of IL-13 markedly inhibits allergen-induced AHR, mucus production and eosinophilia. Furthermore, IL-13 delivery to the airway causes all of these effects. IL-13 is thus both necessary and sufficient for experimental models of asthma. However, the IL-13-responsive cells causing these effects have not been identified. Here we show that mice lacking signal transducer and activator of transcription 6 (STAT6) were protected from all pulmonary effects of IL-13. Reconstitution of STAT6 only in epithelial cells was sufficient for IL-13-induced AHR and mucus production in the absence of inflammation, fibrosis or other lung pathology. These results demonstrate the importance of direct effects of IL-13 on epithelial cells in causing two central features of asthma. PMID- 12091881 TI - Gene therapy--work in progress. PMID- 12091882 TI - Advance online publication. PMID- 12091883 TI - Elevated plasma ghrelin levels in Prader Willi syndrome. PMID- 12091880 TI - Placental growth factor reconstitutes hematopoiesis by recruiting VEGFR1(+) stem cells from bone-marrow microenvironment. AB - The mechanism by which angiogenic factors recruit bone marrow (BM)-derived quiescent endothelial and hematopoietic stem cells (HSCs) is not known. Here, we report that functional vascular endothelial growth factor receptor-1 (VEGFR1) is expressed on human CD34(+) and mouse Lin(-)Sca-1(+)c-Kit(+) BM-repopulating stem cells, conveying signals for recruitment of HSCs and reconstitution of hematopoiesis. Inhibition of VEGFR1, but not VEGFR2, blocked HSC cell cycling, differentiation and hematopoietic recovery after BM suppression, resulting in the demise of the treated mice. Placental growth factor (PlGF), which signals through VEGFR1, restored early and late phases of hematopoiesis following BM suppression. PlGF enhanced early phases of BM recovery directly through rapid chemotaxis of VEGFR1(+) BM-repopulating and progenitor cells. The late phase of hematopoietic recovery was driven by PlGF-induced upregulation of matrix metalloproteinase-9, mediating the release of soluble Kit ligand. Thus, PlGF promotes recruitment of VEGFR1(+) HSCs from a quiescent to a proliferative BM microenvironment, favoring differentiation, mobilization and reconstitution of hematopoiesis. PMID- 12091884 TI - Next steps on ART. PMID- 12091885 TI - WHO and industry combine to form ethics body. PMID- 12091886 TI - Ebola vaccine gets corporate backer. PMID- 12091889 TI - What is translational research? PMID- 12091890 TI - Chris Walsh. PMID- 12091891 TI - Antiretroviral guidelines for resource-limited settings: the WHO's public health approach. PMID- 12091892 TI - Antiretroviral therapy to treat and prevent HIV/AIDS in resource-poor settings. PMID- 12091894 TI - Hemoglobin as a chariot for NO bioactivity. PMID- 12091895 TI - RNA interference meets HIV-1: will silence be golden? PMID- 12091896 TI - Efp: a ring of independence? PMID- 12091897 TI - Extra Rabs unsnarl a lipid traffic jam. PMID- 12091898 TI - Novel notions on newborn lung disease. PMID- 12091899 TI - Subtle? Not this parasite. PMID- 12091901 TI - Heavy functions for light chains. PMID- 12091900 TI - Burning up TNF toxicity for cancer therapy. PMID- 12091902 TI - Degenerate mice. PMID- 12091904 TI - Charting HIV's remarkable voyage through the cell: Basic science as a passport to future therapy. AB - Adequate control of HIV requires impairing the infection, replication and spread of the virus, no small task given the extraordinary capacity of HIV to exploit the cell's molecular machinery in the course of infection. Understanding the dynamic interplay of host cell and virus is essential to the effort to eradicate HIV. PMID- 12091905 TI - Staphylococcus aureus extracellular adherence protein serves as anti-inflammatory factor by inhibiting the recruitment of host leukocytes. AB - Staphylococcus aureus is a human pathogen that secretes proteins that contribute to bacterial colonization. Here we describe the extracellular adherence protein (Eap) as a novel anti-inflammatory factor that inhibits host leukocyte recruitment. Due to its direct interactions with the host adhesive proteins intercellular adhesion molecule 1 (ICAM-1), fibrinogen or vitronectin, Eap disrupted beta(2)-integrin and urokinase receptor mediated leukocyte adhesion in vitro. Whereas Eap-expressing S. aureus induced a 2 3-fold lower neutrophil recruitment in bacterial peritonitis in mice as compared with an Eap-negative strain, isolated Eap prevented beta(2)-integrin-dependent neutrophil recruitment in a mouse model of acute thioglycollate-induced peritonitis. Thus, the specific interactions with ICAM-1 and extracellular matrix proteins render Eap a potent anti-inflammatory factor, which may serve as a new therapeutic substance to block leukocyte extravasation in patients with hyperinflammatory pathologies. PMID- 12091906 TI - The t(8;21) fusion protein, AML1 ETO, specifically represses the transcription of the p14(ARF) tumor suppressor in acute myeloid leukemia. AB - The t(8;21) is one of the most frequent chromosomal translocations associated with acute leukemia. This translocation creates a fusion protein consisting of the acute myeloid leukemia-1 transcription factor and the eight-twenty-one corepressor (AML1 ETO), which represses transcription through AML1 (RUNX1) DNA binding sites and immortalizes hematopoietic progenitor cells. We have identified the p14(ARF) tumor suppressor, a mediator of the p53 oncogene checkpoint, as a direct transcriptional target of AML1 ETO. AML1 ETO repressed the p14(ARF) promoter and reduced endogenous levels of p14(ARF) expression in multiple cell types. In contrast, AML1 stimulated p14(ARF) expression and induced phenotypes consistent with cellular senescence. Chromatin immunoprecipitation assays demonstrated that AML1 ETO was specifically bound to the p14(ARF) promoter. In acute myeloid leukemia samples containing the t(8;21), levels of p14(ARF) mRNA were markedly lower when compared with other acute myeloid leukemias lacking this translocation. Repression of p14(ARF) may explain why p53 is not mutated in t(8;21)-containing leukemias and suggests that p14(ARF) is an important tumor suppressor in a large number of human leukemias. PMID- 12091907 TI - Fluorescence molecular tomography resolves protease activity in vivo. AB - Systematic efforts are under way to develop novel technologies that would allow molecular sensing in intact organisms in vivo. Using near-infrared fluorescent molecular beacons and inversion techniques that take into account the diffuse nature of photon propagation in tissue, we were able to obtain three-dimensional in vivo images of a protease in orthopic gliomas. We demonstrate that enzyme activatable fluorochromes can be detected with high positional accuracy in deep tissues, that molecular specificities of different beacons towards enzymes can be resolved and that tomography of beacon activation is linearly related to enzyme concentration. The tomographic imaging method offers a range of new capabilities for studying biological function; for example, identifying molecular-expression patterns by multispectral imaging or continuously monitoring the efficacy of therapeutic drugs. PMID- 12091909 TI - Uncontrolled C3 activation causes membranoproliferative glomerulonephritis in mice deficient in complement factor H. AB - The alternative pathway of complement is activated continuously in vivo through the C3 'tick-over' pathway. This pathway is triggered by the hydrolysis of C3, resulting in the formation of C3 convertase. This, in turn, generates C3b, which mediates many of the biological functions of complement. Factor H, the main regulator of this activation, prevents formation and promotes dissociation of the C3 convertase enzyme, and, together with factor I, mediates the proteolytic inactivation of C3b. Factor H deficiency, described in 29 individuals from 12 families and in pigs, allows unhindered activation of fluid-phase C3 and severe depletion of plasma C3 (ref. 11). Membranoproliferative glomerulonephritis (MPGN) occurs in factor H-deficient humans and pigs. Although MPGN has been reported in other conditions in which uncontrolled activation of C3 occurs, the role of C3 dysregulation in the pathogenesis of MPGN is not understood. Here we show that mice deficient in factor H (Cfh(-/-) mice) develop MPGN spontaneously and are hypersensitive to developing renal injury caused by immune complexes. Introducing a second mutation in the gene encoding complement factor B, which prevents C3 turnover in vivo, obviates the phenotype of Cfh(-/-) mice. Thus, uncontrolled C3 activation in vivo is essential for the development of MPGN associated with deficiency of factor H. PMID- 12091910 TI - Urocortin-deficient mice show hearing impairment and increased anxiety-like behavior. AB - Urocortin is a member of the corticotropin-releasing hormone peptide family and is found in many discrete brain regions. The distinct expression pattern of urocortin suggests that it influences such behaviors as feeding, anxiety and auditory processing. To better define the physiological roles of urocortin, we have generated mice carrying a null mutation of the urocortin gene. Urocortin deficient mice have normal basal feeding behavior and stress responses, but show heightened anxiety-like behaviors in the elevated plus maze and open-field tests. In addition, hearing is impaired in the mutant mice at the level of the inner ear, suggesting that urocortin is involved in the normal development of cochlear sensory-cell function. These results provide the first example of a function for any peptidergic system in hearing. PMID- 12091912 TI - Complement in glomerulonephritis. PMID- 12091911 TI - Meiotic arrest and aneuploidy in MLH3-deficient mice. AB - MutL homolog 3 (Mlh3) is a member of a family of proteins conserved during evolution and having dual roles in DNA mismatch repair and meiosis. The pathway in eukaryotes consists of the DNA-binding components, which are the homologs of the bacterial MutS protein (MSH 2 6), and the MutL homologs, which bind to the MutS homologs and are essential for the repair process. Three of the six homologs of MutS that function in these processes, Msh2, Msh3 and Msh6, are involved in the mismatch repair of mutations, frameshifts and replication errors, and two others, Msh4 and Msh5, have specific roles in meiosis. Of the four MutL homologs, Mlh1, Mlh3, Pms1 and Pms2, three are involved in mismatch repair and at least two, Pms2 and Mlh1, are essential for meiotic progression in both yeast and mice. To assess the role of Mlh3 in mammalian meiosis, we have generated and characterized Mlh3(-/-) mice. Here we show that Mlh3(-/-) mice are viable but sterile. Mlh3 is required for Mlh1 binding to meiotic chromosomes and localizes to meiotic chromosomes from the mid pachynema stage of prophase I. Mlh3(-/-) spermatocytes reach metaphase before succumbing to apoptosis, but oocytes fail to complete meiosis I after fertilization. Our results show that Mlh3 has an essential and distinct role in mammalian meiosis. PMID- 12091913 TI - Neutrophil chemotaxis in linear and complex gradients of interleukin-8 formed in a microfabricated device. AB - Although a wealth of knowledge about chemotaxis has accumulated in the past 40 years, these studies have been hampered by the inability of researchers to generate simple linear gradients instantaneously and to maintain them at steady state. Here we describe a device microfabricated by soft lithography and consisting of a network of microfluidic channels that can generate spatially and temporally controlled gradients of chemotactic factors. When human neutrophils are positioned within a microchannel, their migration in simple and complex interleukin-8 (IL-8) gradients can be tested. The cells exhibit strong directional migration toward increasing concentrations of IL-8 in linear gradients. Neutrophil migration halts abruptly when cells encounter a sudden drop in the chemoattractant concentration to zero ("cliff" gradient). When neutrophils are challenged with a gradual increase and decrease in chemoattractant ("hill" gradient), however, the cells traverse the crest of maximum concentration and migrate further before reversing direction. The technique described in this paper provides a robust method to investigate migratory cells under a variety of conditions not accessible to study by earlier techniques. PMID- 12091914 TI - Proteomic profiling of mechanistically distinct enzyme classes using a common chemotype. AB - Proteomics research requires methods to characterize the expression and function of proteins in complex mixtures. Toward this end, chemical probes that incorporate known affinity labeling agents have facilitated the activity-based profiling of certain enzyme families. To accelerate the discovery of proteomics probes for enzyme classes lacking cognate affinity labels, we describe here a combinatorial strategy. Members of a probe library bearing a sulfonate ester chemotype were screened against complex proteomes for activity-dependent protein reactivity, resulting in the labeling of at least six mechanistically distinct enzyme classes. Surprisingly, none of these enzymes represented targets of previously described proteomics probes. The sulfonate library was used to identify an omega-class glutathione S-transferase whose activity was upregulated in invasive human breast cancer lines. These results indicate that activity-based probes compatible with whole-proteome analysis can be developed for numerous enzyme classes and applied to identify enzymes associated with discrete pathological states. PMID- 12091915 TI - Genome-wide internal tagging of bacterial exported proteins. AB - As a result of the explosive growth of bacterial genomic and postgenomic information, there is a pressing need for efficient, inexpensive strategies for characterizing the in vivo behavior and function of newly identified gene products. We describe here an internal tagging procedure, based on transposon technology, to facilitate the analysis of membrane-bound and secreted proteins in Gram-negative bacteria. The technique is based on a broad host range transposon (ISphoA/hah), which may be used to generate both alkaline phosphatase (AP) gene fusions and 63-codon in-frame insertions in the genome. The 63-codon insertion encodes an influenza hemagglutinin epitope and a hexahistidine sequence, permitting sensitive detection and metal affinity purification of tagged proteins. For each gene targeted, it is thus possible to monitor the disruption of phenotype (using the transposon insertion), the gene's transcription and translation (using the AP reporter activity), and the behavior of the unfused protein (using the internal tag). Studies on a sequence-defined collection of Escherichia coli strains generated using the transposon showed that the synthesis and subcellular localization of tagged proteins could be readily monitored. The use of ISphoA/hah should provide a cost-effective approach for genome-wide in vivo studies of the behavior of exported proteins in a number of bacterial species. PMID- 12091917 TI - Recombinant DNA. PMID- 12091916 TI - Harnessing microbially generated power on the seafloor. AB - In many marine environments, a voltage gradient exists across the water sediment interface resulting from sedimentary microbial activity. Here we show that a fuel cell consisting of an anode embedded in marine sediment and a cathode in overlying seawater can use this voltage gradient to generate electrical power in situ. Fuel cells of this design generated sustained power in a boat basin carved into a salt marsh near Tuckerton, New Jersey, and in the Yaquina Bay Estuary near Newport, Oregon. Retrieval and analysis of the Tuckerton fuel cell indicates that power generation results from at least two anode reactions: oxidation of sediment sulfide (a by-product of microbial oxidation of sedimentary organic carbon) and oxidation of sedimentary organic carbon catalyzed by microorganisms colonizing the anode. These results demonstrate in real marine environments a new form of power generation that uses an immense, renewable energy reservoir (sedimentary organic carbon) and has near-immediate application. PMID- 12091918 TI - Sources in Orthodox Christianity for bioethical decision-making. PMID- 12091919 TI - Consent and organ donation. PMID- 12091920 TI - The evolution of modern bioethics: the uncertain nature of progress. PMID- 12091921 TI - The constitutionality of court-ordered cesarean surgery: a threshold question. PMID- 12091923 TI - Ethical issues in organ procurement in Chinese societies. PMID- 12091922 TI - A comparative analysis of the right to die in the Netherlands and the United States after Cruzan: reassessing the right of self-determination. PMID- 12091924 TI - The plight of the genetically handicapped newborn: a comparative analysis. PMID- 12091925 TI - Is medical ethics unique? PMID- 12091926 TI - The right of privacy and restraints on abortion under the "undue burden" test: a jurisprudential comparison of Planned Parenthood v. Casey with European practice and Italian law. PMID- 12091927 TI - Malpractice: questions of consent in connection with treatment of genital or urinary organs. PMID- 12091928 TI - Validity of statutes authorizing asexualization or sterilization of criminals or mental defectives. PMID- 12091929 TI - Power of parent to have mentally defective child sterilized. PMID- 12091931 TI - Power of court or other public agency to order medical treatment over parental religious objections for child whose life is not immediately endangered. PMID- 12091930 TI - Tort liability for wrongfully causing one to be born. PMID- 12091932 TI - Physician-patient privilege as applied to physician's testimony concerning wound required to be reported to public authority. PMID- 12091933 TI - Modern status of views as to general measure of physician's duty to inform patient of risks of proposed treatment. PMID- 12091934 TI - Right to maintain action or to recover damages for death of unborn child. PMID- 12091935 TI - Jurisdiction of court to permit sterilization of mentally defective person in absence of specific statutory authority. PMID- 12091936 TI - From experience to principles: method in clinical and research ethics. PMID- 12091937 TI - Can bioethics be both rigorous and practical? PMID- 12091938 TI - The clinical teaching of medical ethics. PMID- 12091939 TI - Beyond the Hippocratic Oath: bioethics and law. PMID- 12091940 TI - New officers elected at Augusta meeting; position paper on health care and society adopted. PMID- 12091941 TI - "Hippocrates rises anew": prominent ethicians and physicians issue A.D. 1995 restatement of the Oath of Hippocrates (circa 400 B.C. PMID- 12091942 TI - What's narrative ethics? PMID- 12091943 TI - Helping the newborn via genetic counseling. PMID- 12091944 TI - Institutional animal care and use committees and the moderate position. PMID- 12091945 TI - Animal models in 'exemplary' medical research: diabetes as a case study. PMID- 12091946 TI - Moral issues associated with bioengineered species: stewardship, abuse and sustainability. PMID- 12091947 TI - On the genetic manipulation of animals. PMID- 12091948 TI - Experimenting on animals: a reconsideration. PMID- 12091949 TI - The responsible use of animals in biomedical research. PMID- 12091950 TI - Should we genetically engineer hogs? PMID- 12091951 TI - Response: of pigs and primitive notions. PMID- 12091952 TI - Test-tube babies: for or against? PMID- 12091953 TI - Gene splicing: fact and fantasy. PMID- 12091954 TI - Some comments on the Abortion and Sterilization Act 2 of 1975 after one year's operation: legal aspects. PMID- 12091955 TI - The Abortion and Sterilization Act 2 of 1975: a third opinion (Part 1). PMID- 12091956 TI - The Abortion and Sterilization Act 2 of 1975: a third opinion (part II). PMID- 12091957 TI - Acquired immune deficiency syndrome (AIDS) in the workplace: guidelines for AIDS information and education and for personnel management issues. PMID- 12091959 TI - Abortion and the double effect principle. PMID- 12091958 TI - The poet, the biographer and the shrink: psychiatrist-patient confidentiality and the Anne Sexton biography. PMID- 12091960 TI - The Nancy Cruzan case. PMID- 12091961 TI - Breaking boundaries: AIDS and social justice in Australia. PMID- 12091962 TI - The treatment of AIDS behind the walls of correctional facilities. PMID- 12091963 TI - Medicare coverage for diabetes care. PMID- 12091964 TI - Cardiac testing: what to expect. PMID- 12091965 TI - Cooking from the Diabetes Food Pyramid. Vegetables. PMID- 12091966 TI - Taking to the air with diabetes. PMID- 12091967 TI - Dining out with confidence. PMID- 12091968 TI - For parents. Sending your kid to camp. PMID- 12091969 TI - Baja light. Making the most of Mexican cuisine. PMID- 12091970 TI - Using food records for better control. PMID- 12091971 TI - Treatment options for type 2 diabetes. Finding what's best for you. PMID- 12091972 TI - Supermarket smarts. Packaged cookies. PMID- 12091973 TI - Diabetes quiz. How much do you know about sweeteners? PMID- 12091974 TI - Sports & fitness. Planning an active summer. PMID- 12091975 TI - [Cross reactions to curare agents and prediction of their safe use]. PMID- 12091976 TI - [Skin tests for diagnosis of allergies to local anesthetics and antibiotics]. PMID- 12091977 TI - [Skin tests for diagnosis of hypersensitivity to colloidal plasma substitutes]. PMID- 12091978 TI - [Risk factors for latex allergy. Diagnostic methods for aprotinin allergy]. PMID- 12091979 TI - [Are there risk factors of allergic reactions related to patient factors, to drugs, techniques of use? Predictive indications]. PMID- 12091980 TI - [Can prevention of allergic risk be assured with preanesthetic medication]. PMID- 12091981 TI - [Management of anaphylactoid reactions associated with anesthesia, and treatment of anaphylactic shock]. PMID- 12091982 TI - [The new recommendations: why comments?]. PMID- 12091983 TI - [Survey on prevention of anaphylaxis]. PMID- 12091984 TI - [Case records and jurisprudence]. PMID- 12091985 TI - [Pharmacovigilance of anaphylactic shock from anesthetics over a 6 year period (from January 1994 to December 1999]. PMID- 12091987 TI - [Methodology for reducing risk of anaphylaxis in anesthesia]. PMID- 12091986 TI - [What is the real risk of drug hypersensitivity in anesthesia? Incidence. Clinical aspects. Morbidity-mortality. Substances responsible]. PMID- 12091988 TI - [What are the physiopathological mechanisms? How can severe anaphylactoid reactions be explained]. PMID- 12091989 TI - [Comments on diagnosis of anphylactoid reactions. Diagnostic tests in vitro]. PMID- 12091990 TI - Reducing the risk of anaphylaxis during anaesthesia. Abbreviated text. PMID- 12091991 TI - [Skin tests for diagnosis of curare allergy]. PMID- 12091992 TI - Nasopharyngeal glioma in a new-born girl. AB - Nasal gliomas are uncommon tumours of neurogenic origin that occur sporadically. They are diagnosed with MRI and a preoperative biopsy, and surgery is the treatment of choice. Most of the gliomas emerge from the nasal cavity, but only a few cases of nasopharyngeal gliomas have been reported. We present one case of a nasopharyngeal glioma and two cases of nasal gliomas. PMID- 12091994 TI - Dacryocystorhinostomy. Diagnosis and treatment of nasolacrimal canal obstructions. AB - Since Toti's original description of the dacryocystorhinostomy (CDR) in 1904, most surgery for relief of lacrimal obstruction has been performed through an external incision. Although the endonasal approach was for the first time introduced by Caldwell in 1893, its use stayed limited due to difficulties in visualizing the endonasal structures during the operation. The advance of the operating microscope and later a rigid endoscope into the surgery awakened interest for the endonasal approach. Endonasal DCR surgery has the advantages of preventing a scar on the skin and preserving the pump function of the naslacrimal sac. If the surgeon is experienced in endoscopic or microscopic surgery, correct localization of the window and addressing possible other nasal pathologies is possible in the same session. The endonasal laser DCR is not the first operation to be preferred due to the high cost, longer operation time, and less satisfactory results. It may be an alternative in cases with a tendency to bleeding. Endonasal surgery can not help canalicular problems. In these cases external surgery should be preferred. In revision cases, the endonasal approach is preferred by almost all authors. Even in external surgery, endoscopy may accompany the external approach to check the site of the fistula. Silicone intubation may be used routinely for two months to have better results, but no longer than 3 months to avoid complications. Silicone intubation is especially recommended in canalicular stenosis, small scarred lacrimal sacs and in reoperations. In this review article, diagnostic tests and various surgical methods, the advantages and disadvantages, were compared. Reasons for failure after surgery are also discussed. PMID- 12091993 TI - Early depictions of the nasal turbinates in the 15th century. AB - Although nasal turbinates had already been described by Hippocrates, it was not until the 15th century that they were depicted. The inferior turbinate was shown for the first time rather true- to nature in the works of the Middle Rhenic Master circa 1450-1460 and Leonardo da Vinci in 1489. The posterior ends of the middle turbinates were depicted on a woodcut by Georg Thomas for Dryander's "Anatomiae ... pars prior" in 1536. These and a few other examples show that some artists were ahead of the medical profession in demonstrating anatomical details in the 15th and 16th century. PMID- 12091995 TI - Short-term repeatability and correlates of laboratory measures of nasal function in patients with seasonal allergic rhinitis. AB - The purpose of this study was to determine the variability of laboratory nasal function tests in 26 patients (18 female) with seasonal allergic rhinitis (SAR) (mean age 38.1 years). Their usual medication for SAR was withheld for 2 separate one week washout periods, separated by at least 2 weeks, in order to produce clinically significant nasal airflow obstruction. Measurements were made on both occasions for nasal nitric oxide (NO), nasal peak inspiratory flow (nPIF), oral PIF (oPIF), nasal forced inspiratory flow rate in 1 second (nFIV1), oral FIV1 (oFIV1). The respective nasal-oral ratios for FIV1 and PIF were also determined. The intra-individual coefficient of variation was: NO = 14%, nFIV1 = 4%, nFIV1/oFIV1 ratio = 10%, nPIF = 8% and nPIF/oPIF ratio = 12%. Linear regression analysis showed significant (p < 0.05) correlations between nPIF and nFIV1 (R2 = 0.45) and between nPIF/oPIF and nFIV1/oFIV1 (R2 = 0.20). In conclusion, there was a good correlation between the two methods of nasal inspiratory flow, although FIV1 had a lesser degree of variability. PMID- 12091996 TI - Cost analysis of regular and filgrastim treatment in patients with refractory chronic rhinosinusitis. AB - In this double blind randomized placebo controlled clinical trial of filgrastim in chronic sinusitus, we analyzed costs of a 24-week interval in which filgrastim was administered. Since we hypothesized that the scheduled preventive visits within the trial might cause savings as compared to the regular situation in which these patients have a strong tendency to visit the outpatient clinic immediately in case of complications, direct medical costs within the trial were also compared to costs of regular treatment. The difference in costs between both trial groups was driven by the filgrastim costs (Euro 5108). If filgrastim costs were left out of consideration, no significant difference in direct medical costs remained between the filgrastim and placebo groups (Euro 2904 and Euro 2765, respectively). Indirect medical costs also showed no significant differences. Within a regular situation, costs of a 24-week interval were Euro 896. As filgrastim treatment had already been shown not to improve the quality of life, its cost-effectiveness in these patients can only be favourable in case of major clinical improvements. Furthermore, scheduled preventive visits in chronic sinusitis patients appear not to cause savings as compared to the situation in which patients are only seen in case of recurrences. PMID- 12091997 TI - Epidemiological and clinical aspects of nasal polyposis in France; the ORLI group experience. AB - Nasal polyposis (NP) is a common condition in patients consulting ENT practitioners in France. A multicenter prospective study was performed to evaluate symptoms, demography, environmental factors, personal and family history and associated conditions like asthma, and food or drugs sensitivity (FDS) in patients suffering from NP. In each investigation center assessments were performed at the moment of the initial consultation by the same investigator, then updated with complementary exploration results required by the protocol. The chi 2 test and the Fisher test were used for statistical analysis. In this study 224 patients were included. Males were predominant at 63%. Asthma was found in 45% of cases without relevant sex difference. However, FDS, positive in 31% of the patients, was statistically higher in females than in males (42.9% vs. 24.4%). Severe and major symptoms were more frequently found in the female population. Environment and habitat factors did not appear to be relevant. High rates of NP (52.66%) and asthma (43.58%) were found in the family history. Hereditary factors were suggested and lead us to further study the genetic factors potentially involved in this pathology. PMID- 12091998 TI - Incidence of chronic hyperostotic rhinosinusitis in patients undergoing primary sinus surgery compared to revision surgery. AB - HYPOTHESIS: The incidence of chronic hyperostotic rhinosinusitis has been underreported due to poor recognition of the disease process. The surgical management of chronic hyperostotic rhinosinusitis with prolonged pre and post operative antibiotic therapy can alter clinical course of the disease process. STUDY DESIGN: A prospective study of 40 patients based at two teaching tertiary care institutions. MATERIAL AND METHODS: CT scans of twenty patients (ten undergoing primary sinus surgery and ten undergoing revision sinus surgery) were randomly selected from each institution and reviewed by an independent radiologist for evidence of hyperostosis and compared to original reading of the CT scans. RESULTS: The finding of sinus hyperostosis is rarely reported by the radiologist on the sinus CT scan where the focus is always on mucosal disease. The incidence of sinus hyperostosis is higher, in patients undergoing revision sinus surgery than patients undergoing primary sinus surgery. CONCLUSION: Sinus hyperostosis is not an uncommon finding in chronic refractory sinusitis which may require long term intravenous antibiotic therapy in conjunction with surgery to provide symptomatic relief. PMID- 12091999 TI - Excision of rhinophyma with a laser scanner handpiece--a modified technique. AB - Rhinophyma is a slowly progressive, benign dermatological disorder of the tip of the nose. The most widely accepted theory is that rhinophyma is the end stage result of chronic acne rosacea. The primary reason for excision of rhinophyma is cosmetic deformity. However, nasal obstruction may present, as rhinophyma may obstruct nasal vestibules. Surgery is the treatment of choice for rhinophyma. Medical therapy has not given satisfactory results. The surgical treatment modalities are divided into two main groups. The first is complete excision, with primary closure for small lesions, or skin grafting for large lesions. The second group includes incomplete excision followed by re-epithelialization from the remaining glandular epithelium. The carbon dioxide (CO2) laser has been advocated for excision of rhinophyma. Newer instrumentation, such as Swiftlase and SurgiTouch (ESC Sharplan) or Ultrapulse (Coherent) is more effective in vaporization and carbonization resulting in better hemostasis (Ries and Speyer, 1996). In this report, 7 patients with rhinophyma treated with the Swiftlase are reviewed. The surgical technique, the complications and the follow-up are discussed. PMID- 12092000 TI - Fibrin glue treatment for epistaxis. AB - Our study was made to prove the second-generation surgical fibrin sealant Quixil to be an effective substitute for nasal packing, chemical coagulation and cautery in management of patients with epistaxis. Our series includes 204 patients with anterior epistaxis (186), and with posterior epistaxis (18) as results of trauma, clotting disorders, chronic and/or atrophic rhinitis and upper respiratory infections, and hypertension. Patients were randomly divided into four groups: with fibrin glue (67) (Quixil), with electric cautery (61), with silver nitrate coagulation (60), and with foam nasal packing (16). For the fibrin glue group, hemostasis was achieved by spraying with 0.3 ml Quixil fibrin glue to each bleeding nostril. The results were excellent in all of the 62 (92.5%) patients of the fibrin glue group with complete and immediate hemostasis. We found good healing of bleeding sites, no swelling and secondary bleeding, no inflammation, no plaque or crists. Three months monitoring of atrophic changes of the nasal mucosa proved absence of atrophy of the nasal mucosa. In this group, the bleeding time averaged 2 min 30 sec since the moment of admittance. In the groups where cautery, coagulation, or nasal packing was used, we found local swelling, pain, and slow healing of the bleeding site with accidental atrophy of the nasal mucosa. The rates of these side effects were significantly higher in comparison with the fibrin glue group. The bleeding time was also longer. We found that the fibrin glue is more effective hemostatic in comparison with foam nasal packing, cautery and coagulation, and provides no complications usual for these types of treatment of epistaxis. PMID- 12092001 TI - Septal-turbinate-suture in endonasal sinus surgery. AB - The middle turbinate (MT) is an important landmark in sinus surgery with specific functions in the nasal physiology. Because of postoperative instability and corresponding mucosal defects in the middle nasal meatus scar formations between the MT and the lateral nasal wall are frequent. Lateralization and scarring to the lateral nasal wall can be avoided by means of a resorbable mattress suture (septal-turbinate-suture (STS)) through the head of the MT and the septum. PMID- 12092002 TI - Hangman's knot in securing bypass tubes in endonasal dacryocystorhinostomy. AB - We performed 15 cases of dacryocystorhinostomy endoscopically with an 86% success rate. Using a Hangman's knot, the bypass tubes were placed in a secure and lasting way. PMID- 12092003 TI - Endoscopic removal of a nasal septum chondrosarcoma. AB - We describe an alternative, endoscopical technique for the surgical removal of a rare chondrosarcoma on the nasal septum. The technique requires a resection of the whole tumour within margins in healthy osteocartilaginous nasal septum under endoscopic guidance and includes bilateral middle turbinectomy and ethmoidectomy. We present the case of a 57-year-old woman with a well-differentiated chondrosarcoma (Grade I) of the nasal septum. Endoscopical surgery resulted without any cosmetic deformity problems, and the functional result was favourable. No recurrence was evidenced after a follow-up of 3 years. The main advantages of this technique are the excellent functional and cosmetic results without any surgical reconstructive techniques. This approach may provide a minimally invasive method to remove well-differentiated, low-grade (Grade I) and size limited malignant cartilaginous tumours of the nasal septum. PMID- 12092004 TI - Propranolol as an inhibitor of some cytochrome P450-dependent monooxygenase activities in native and induced rat liver microsomes. AB - The in vitro effect of propranolol (10(-3) M and 10(-4) M), a nonselective and extensively metabolized beta-adrenergic blocking agent, on rat liver drug metabolism in native and induced (with phenobarbital and beta-naphthoflavone [beta-NF]) microsomes was studied. The type of inhibition and the inhibitory constants of some cytochrome P450-dependent microsomal enzyme reactions (hexobarbital oxidation [HBO], ethylmorphine-N-demethylation [EMND], aniline hydroxylation [AH], ethoxycoumarin-O-deethylation [ECOD], ethoxyresorufin-O dealkylation [EROD] and penthoxyresorufin-O-dealkylation [PROD]) were estimated. The results showed that propranolol competitively inhibited AH activity in native microsomes. The type of inhibition was changed from competitive to noncompetitive in all other enzyme activities studied. This inhibition was more pronounced after phenobarbital induction in PROD (Ki = 0.11 +/- 0.01 mM), ECOD (Ki = 0.40 +/- 0.09 mM) and EMND (Ki = 0.59 +/- 0.1 mM), and after beta-NF induction in AH (Ki = 0.28 +/- 0.05 mM) and in HBO (Ki = 0.35 +/- 0.1 mM) in native microsomes. It was assumed that the noncompetitive type of inhibition is due to the covalent binding of reactive metabolites derived from propranolol to hepatic microsomal proteins. The competitive type of inhibition of AH suggested a common P450 isoenzyme in the metabolism of propranolol and aniline. Thus, in this study, propranolol has been found to be not only a selective inhibitor of CYP2D6 isoenzyme-dependent reactions, but also a nonspecific inhibitor of other cytochrome P450 isoenzymes. PMID- 12092006 TI - L-arginine reduces structural remodeling in the diabetic rat myocardium. AB - Metabolism, monitored via in situ catalytic enzyme histochemistry and fine structure, was studied in the myocardium of chronic diabetic male Wistar rats administered L-arginine (12.8 mg/100 g/day) for 24 weeks. Diabetes was induced with a single i.v. injection of 55 mg/kg streptozotocin. After 6 months, the tissue of the left ventricle was processed for electron microscope examination and transmural tissue blocks were frozen for enzyme histochemistry. In diabetic myocardium, heterogeneous ischemia-like subcellular alterations of cardiomyocytes and capillaries were observed, together with interstitial fibrosis. This structural remodeling was accompanied by significantly decreased activity of endothelial nitric oxide synthase (NOS) and heterogeneously decreased activities of glycogen phosphorylase (GlPh), hydroxybutyrate dehydrogenase (HBDH) and adenosine triphophatases (ATPases) throughout the myocardium. In arginine-treated diabetic rats, there was evidence of protected structural integrity of endothelial cells and attenuated structural disturbances of cardiomyocytes. This was associated with the markedly preserved histochemical activities of all detected enzymes in comparison with nontreated diabetic rats (NOS 98.7 +/- 10.5% vs. 35.4 +/- 4.1%; ATPases 82.7 +/- 9.1% vs. 69.3 +/- 5.2%; GlPh 65.2 +/- 8.3% vs. 45.5 +/- 3.8%; HBDH 68.9 +/- 8.5% vs. 44.1 +/- 6.7% of control values). The results indicate that long-term supplementation of L-arginine may account for the reduction of diabetes-induced myocardial structural remodeling. PMID- 12092005 TI - Recombinant human endostatin is beneficial to endothelial cell growth exposed to mildly oxidized low-density lipoproteins. AB - Endostatin significantly reduced atherosclerosis in genetically susceptible mice. One of the main factors associated with atherogenesis is oxidized low-density lipoproteins (LDL), which also causes apoptosis of endothelial cells. Therefore, we proposed that the antiatherogenic effect of endostatin was partly associated with its protective effect on the endothelial injury induced by oxidized LDL. To confirm such a hypothesis, we studied the effects of recombinant human endostatin (rhEndo) on the proliferation of cultured endothelial cells exposed to mildly oxidized LDL (mox-LDL), rhEndo did not show an obvious inhibitory effect on the proliferation of rabbit aorta endothelial cells (RAEC) (p > 0.05), while mox-LDL inhibited their proliferation (p < 0.01 or p < 0.05). Interestingly, rhEndo seemed to antagonize the role of mox-LDL in inhibiting the proliferation of RAEC. rhEndo seemed, thus, to be beneficial to the proliferating endothelial cells, suggesting that it protects RAECs from the injury caused by mox-LDL. The activity of rhEndo in endothelial cells may possibly result from the interaction of different factors in cell signaling, which remains to be further elucidated. PMID- 12092008 TI - Infusion of maintenance fluids with glucose (Veen 3G) is superior to maltose infusion (Actit) in the rate of energy utilization in rabbits. AB - In the present study, we examined the rates of urinary excretion of glucose and maltose after an infusion of maintenance fluid with glucose or maltose in adult rabbits. Three maintenance fluids (sugar-free, 5% glucose [Veen 3G] and 5% maltose [Actit]), which contained different sugars but were identical in electrolyte and acetate compositions and concentrations (Na: 45, K: 17, Mg: 5, Cl: 37, H2PO4: 10 and CH3COO: 20 mEq/l), were used in this study. In addition, the optimum infusion speed for maintenance therapy (10 ml/kg/h) was used. Animals were not given food or water during the 10-day period of administration. The body weights of the animals were measured every day. The concentrations of total protein, albumin, free fatty acids and glucose in the serum were measured. Urine samples for determination of glucose and maltose concentrations were collected from the 1st to 10th administrations. After infusion with 5% maltose, urinary maltose excretion decreased time-dependently, while that of glucose increased. This suggests that maltase activity time-dependently increases after infusion with maltose. In addition, total sugar was only minimally excreted into urine in the 5% glucose group compared with the 5% maltose group. Thus, the glucose infusion was superior to the maltose infusion in the rate of energy utilization. However, neither the loss of body weight nor the increase in concentration of free fatty acids in serum differed significantly among the 3 groups. In conclusion, infusion of maintenance fluid with 5% maltose results in the excretion of maltose and glucose into urine, since enzymatic hydrolysis of maltose to glucose is limited to that by maltase. PMID- 12092007 TI - Radioprotective effects of exogenous glutathione against whole-body gamma-ray irradiation: age- and gender-related changes in malondialdehyde levels, superoxide dismutase and catalase activities in rat liver. AB - Age- and gender-related changes in malondialdehyde (MDA) levels, superoxide dismutase (SOD) and catalase (CAT) activities in rat livers exposed to different doses of whole-body gamma-ray radiation were determined. In addition, the effects of exogenous glutathione (GSH) against radiation injury in rat livers were investigated. We found that MDA levels have an age-associated increment and an increasing radiation dose-related elevation, although they decrease slightly in the 4 Gy group. The MDA levels in old rats were lower in males than in females, while those of young rats did not change. There were no observed age-related changes in SOD activities, although male rats had higher SOD activity than females. Female rats had the highest CAT activities in the 4 Gy group, while male rats had the highest CAT activities in the 6 Gy group. CAT activities in the 8 Gy group were lower than those of the 2 Gy group for each gender and age. While MDA levels were decreased and CAT activities increased by GSH, SOD activities remained unchanged. The results indicate that gamma-ray radiation affects gender- and age-dependent MDA levels, SOD and CAT activities. Administration of GSH appears to be a useful approach to reduce radiation injury by reducing MDA levels and increasing CAT activities. PMID- 12092009 TI - Gateways to clinical trials. AB - Gateways to Clinical Trials is a guide to the most recent clinical trials in current literature and congresses. The data in the following tables can be retrieved from the Clinical Studies knowledge area of Prous Science Integrity, the drug discovery and development portal, http://integrity.prous.com. This issue focuses on the following selection of drugs: Abacavir sulfate, abarelix, abciximab, acarbose, alefacept, alteplase, amisulpride, amoxicillin trihydrate, apomorphine hydrochloride, aprepitant, argatroban monohydrate, aspirin, atenolol; Betamethasone dipropionate, betamethasone valerate, bicalutamide, bleomycin sulfate; Calcium carbonate, candesartan cilexetil, celecoxib, cetirizine hydrochloride, cisplatin, clarithromycin, clavulanate potassium, clomethiazole edisilate, clopidogrel hydrogensulfate, cyclophosphamide, chorionic gonadotropin (human); Dalteparin sodium, desloratadine, dexamethasone, doxorubicin, DPC-083; Efalizumab, efavirenz, enoxaparin sodium, eprosartan mesilate, etanercept, etoposide, ezetimibe; Faropenem daloxate, fenofibrate, fluocinolone acetonide, flutamide, fluvastatin sodium, follitropin beta, fondaparinux sodium; Gabapentin, glibenclamide, goserelin, granisetron hydrochloride; Haloperidol, hydrochlorothiazide; Imiquimod, interferon beta-1a, irbesartan, iseganan hydrochloride; L-758298, lamivudine, lanoteplase, leflunomide, leuprorelin acetate, loratadine, losartan potassium; Melagatran, metformin hydrochloride, methotrexate, metronidazole, micafungin sodium, mitoxantrone hydrochloride; Nelfinavir mesilate, neutral insulin injection, nizatidine; Olopatadine hydrochloride, omeprazole, ondansetron hydrochloride; Pamidronate sodium, paracetamol, paroxetine hydrochloride, perindopril, pimecrolimus, pioglitazone hydrochloride, piroxicam, pleconaril, pralmorelin, pravastatin sodium, prednisolone, prednisone, propofol; Raloxifene hydrochloride, ranpirnase, remifentanil hydrochloride, risedronate sodium, risperidone, rofecoxib, ropinirole hydrochloride, rosuvastatin calcium; Sevoflurane, sildenafil citrate, simvastatin, somatropin; Tacrolimus, tamoxifen citrate, telmisartan, temozolomide, thiopental sodium, tinzaparin sodium, tirofiban hydrochloride, treosulfan, triamcinolone acetonide; Urokinase; Valsartan, vardenafil, vincristine; Warfarin sodium; Ximelagatran; Zidovudine. PMID- 12092010 TI - Gestational trophoblastic neoplasia and evidence-based medicine. AB - OBJECTIVE: To determine whether protocols for the management of gestational trophoblastic neoplasia conform to the principles of evidence-based medicine. STUDY DESIGN: Protocols for the management of low- and high-risk gestational trophoblastic neoplasia were examined to determine to what extent they conformed to the principles of evidence-based medicine. RESULTS: Nearly all current chemotherapy regimens for gestational trophoblastic neoplasia are based on the experience of management of various risk groups, variously defined. Some prospective, randomized studies were flawed by faulty selection criteria. Local population variations may influence the results of management. CONCLUSION: The management of trophoblastic neoplasia is based on physician experience. Nearly all prospective, randomized studies have been flawed. There is a need for carefully planned prospective studies with stringent inclusion criteria to determine the most effective and cost-effective and least toxic therapy, particularly for low-risk neoplasia. PMID- 12092011 TI - Detection of hCG in trophoblastic disease. The USA hCG reference service experience. AB - hCG is a glycoprotein hormone composed of two dissimilar subunits. This hormone is not only heterogeneous in peptide structure but also in combination of subunits and carbohydrate structure. Common hCG-related molecules include hCG, hyperglycosylated hCG, nicked hCG, hCG missing the beta-subunit C-terminal peptide, free alpha-subunit, free beta-subunit, nicked free b-subunit and urine beta-core fragment. This article discusses the structures these hCG-related molecules and their occurrences in early pregnancy, 7-week to term pregnancy, hydatidiform mole (preevacuation and postevacuation), persistent gestational trophoblastic disease, choriocarcinoma and other malignancies. Multiple serum hCG tests are evaluated, and their abilities to detect the multiple hCG-related molecules are investigated. The accuracy of different serum hCG tests in detecting hCG and hCG-related molecules in patients with gestational trophoblastic diseases is evaluated. The findings of persistent low hCG values in the absence of pregnancy or an identifiable malignancy are examined. In addition, the false positive hCG assay problem is discussed. False positive hCG tests have led to many incidences in which gestational trophoblastic disease has been erroneously diagnosed and needlessly treated. hCG tests are identified that give a disproportionate number of false positive results. Finally, guidelines are presented for selecting an hCG test. PMID- 12092012 TI - Negotiating a staging and risk factor scoring system for gestational trophoblastic neoplasia. A progress report. AB - OBJECTIVE: To discuss the current problems in establishing a unified staging and risk factor scoring system for trophoblastic disease under the auspices of the International Federation of Gynecology and Obstetrics (FIGO). STUDY DESIGN: To present a history of the classifications of trophoblastic disease and compare these with the proposals presented to FIGO by the International Society for the Study of Trophoblastic Disease, the International Gynecologic Cancer Society and the Society of Gynecologic Oncologists. RESULTS: The situation concerning the proposed 2000 FIGO trophoblast classification was resolved by FIGO's acceptance of the recommendations outlined in this paper. CONCLUSION: There is a scientific and clinical need for FIGO to adopt the recommendations of the majority of physicians with expertise in treating trophoblastic disease for a rational classification of trophoblastic disease. It is gratifying that FIGO has done so. PMID- 12092013 TI - Advances in management of high-risk gestational trophoblastic tumors. AB - Multimodality therapy with combination chemotherapy employing etoposide, high dose methotrexate, actinomycin D, cyclophosphamide and vincristine and adjuvant radiotherapy and surgery, when indicated, has resulted in cure rates of 80-90% in patients with high-risk metastatic gestational trophoblastic tumors. However, 25 30% of high-risk patients will have an incomplete response to first-line chemotherapy or will relapse from remission. Most of these patients will have a clinicopathologic diagnosis of choriocarcinoma, multiple metastases to sites other than the lung and vagina, and failed or inappropriate previous chemotherapy, resulting in very high World Health Organization scores. Salvage chemotherapy with cisplatin/etoposide, usually in conjunction with bleomycin or ifosfamide, as well as surgical resection of sites of resistant disease in selected patients, will result in a cure for most patients. Colony-stimulating factors should be used to prevent treatment delays and dose reductions. Newer anticancer agents, such as paclitaxel and gemcitibine, and high-dose chemotherapy with or without autologous bone marrow transplantation or peripheral blood stem cell support may play a role in the future management of selected patients. PMID- 12092014 TI - Twenty-five years' clinical experience with placental site trophoblastic tumors. AB - OBJECTIVE: To describe 34 cases of placental site trophoblastic tumor (PSTT) treated at Charing Cross Hospital over 25 years. STUDY DESIGN: Between 1975 and 2001, 1,685 patients with gestational trophoblastic disease (GTD) were treated; 34 of them had PSTT (2%). The computer database clinical notes and the pathology reports were accessed to obtain data on this patient group. The data were subsequently analyzed using Excel computer software. RESULTS: The mean age of the group was 33 years (95% CI 25-41). The antecedent pregnancy was a full-term, normal one in 18 cases (53%), a molar pregnancy in 7 (21%) and a missed abortion in 5 (15%). The mean interval from the last pregnancy to diagnosis was 3.4 years (95% CI 1.9-4.9). The range of serum hCG concentrations at diagnosis was 0 58,000, 79% with levels < 1,000 and 58% < 500. hCG was raised in all with active disease. The most frequent presenting complaint was vaginal bleeding, in 27 cases (79%). At diagnosis, the disease was localized to the uterus in 15 (44%); there was pelvic involvement in 8 (24%) and lung secondaries in 10 (29%). All seven deaths were disease related (21%); all had lung secondaries and presented more than four years since the last pregnancy. Excluding the seven deaths, the primary treatment was surgery alone in 10 cases (37%) (8 hysterectomies and 2 dilatation and curettages); 4 had surgery followed by adjuvant chemotherapy; 5 had neoadjuvant chemotherapy followed by surgery; 1 had chemotherapy alone, and the disease recurred and was successfully rechallenged; and 5 had surgery between chemotherapy cycles. The most common regimens consisted of EMA/CO and EP/EMA. CONCLUSION: Risk factors for death include lung metastatic involvement (50%) and an antecedent pregnancy interval of four years or more (100%). In contrast, those with no extrapelvic disease or a pregnancy interval of less than four years had 100% survival. In two-thirds of patients with disease limited to the uterus, surgery alone was curative. The WHO scoring system for GTD did not correlate with this outcome. Patients with PSTT should be managed separately from those with other types of GTD, as the disease behavior is different. PMID- 12092015 TI - Management of brain metastases in patients with high-risk gestational trophoblastic tumors. AB - OBJECTIVE: To analyze the results of current treatment of patients with brain metastases from high-risk gestational trophoblastic tumors. STUDY DESIGN: Consecutive patients treated between June 1981 and the end of 2000 with brain metastases from high-risk gestational trophoblastic tumors were selected from our computerized database. RESULTS: There were 39 patients with cerebral metastases from high-risk gestational trophoblastic tumors, and 30 (79.5%) of these patients are alive and in remission. Four patients died within 8 days of admission from disease extent. If these four patients are excluded, the survival of the remaining 35 patients is 86%. Eight patients had received prior chemotherapy, and 3 died of the disease. The antecedent pregnancy (AP) was term delivery in 23 (59%), and in 2 of those patients there was a prior history of a molar pregnancy in an AP. Six patients had a history of molar pregnancy as the AP, and in 10 the type of AP was uncertain. The presence of both liver and brain metastases was a particularly adverse prognostic combination, and only one of five patients is still alive in remission. No deaths or relapses occurred beyond 30+ months from the initiation of high-dose etoposide, methotrexate and actinomycin D with cyclophosphamide and vincristine chemotherapy. CONCLUSION: With appropriate management, the outlook for patients with brain metastases from high-risk gestational trophoblastic tumors is good, and the majority of patients achieved sustained remission and probably a cure with chemotherapy as the dominant form of treatment. When the tumor is sufficiently chemosensitive, the blood-brain barrier does not prevent disease elimination. PMID- 12092016 TI - Hypertension in pregnancy and preeclampsia. Knowledge and clinical practice among obstetrician-gynecologists. AB - OBJECTIVE: To examine the knowledge and practice patterns of obstetrician gynecologists concerning management of hypertensive disorders of pregnancy. STUDY DESIGN: Surveys were mailed to 1,116 fellows of the American College of Obstetrics and Gynecology; 416 of them constituted the Collaborative Ambulatory Research Network. Seven hundred more were chosen at random. The survey contained questions on physician and patient demography and on knowledge and practice patterns concerning management of various hypertensive disorders during pregnancy. RESULTS: A total of 401 completed surveys were analyzed. There was no difference between respondents and nonrespondents in either sex ratio (P = .410) or age (46.9 +/- 0.4 versus 48.1 +/- 0.4 years, P = .131). Most respondents (84.5%) would manage mild preeclampsia on an outpatient basis, and most (58.6%) usually managed preeclampsia independently. There was considerable variation in clinical practice. For example, about one of four respondents (27.4%) do not use seizure prophylaxis during labor in mild preeclampsia. Among physicians who do utilize magnesium sulfate for seizure prophylaxis, the mean standard loading dose was 4.5 +/- .1 g intravenously. More than half the respondents (54.9%) would employ preeclampsia prevention procedures. Most respondents (74.6%) said that there is a role for management of severe preeclampsia remote from term. Intrauterine growth restriction would be used as an indication for immediate delivery by 60.6% of respondents; female physicians were more likely to use intrauterine growth retardation as an indication for immediate delivery (chi 2 = 5.7, P = .017). PMID- 12092017 TI - Traditional care of the perineum during birth. A prospective, randomized, multicenter study of 1,076 women. AB - OBJECTIVE: To investigate the influence of the traditional hands-on versus the innovative hands-poised method on the risk of perineal trauma during vaginal delivery and on neonatal outcomes. STUDY DESIGN: In a prospective, randomized, multicenter study, 1,161 of 1,505 women giving birth at the Departments of Obstetrics and Gynecology of the University Hospital of Vienna and Semmelweis Women's Hospital, Vienna, between February and September 1999, were randomized into the trial. In the hands-on method, the left hand of the midwife puts pressure on the infant's head, and the right hand is placed against the perineum. In the hands-poised method, the midwife guides the parturient through the birth without touching the perineum, prepared to apply light pressure on the infant's head. RESULTS: One hundred eighty-seven of 574 women (32.5%) in the hands-on group and 180 of 502 women (35.8%) in the hands-poised group experienced perineal tears (P = .5). Sixteen women (2.7%) treated with the hands-on method developed third-degree perineal tears as compared with five women (0.9%) treated with the hands-poised method (P < .05). In the hands-on group, 103 women (17.9%) underwent episiotomy as compared with 51 cases (10.1%) in the hands-poised group (P < .01). No significant differences in neonatal outcomes were observed between the two groups. CONCLUSION: Our data suggest that a policy of hands-poised care is more suitable for preserving the perineum during birth and is a safe and effective birthing alternative for women. PMID- 12092018 TI - Failed methotrexate treatment of cervical pregnancy. Predictive factors. AB - OBJECTIVE: To determine the risk factors when primary methotrexate treatment of cervical pregnancy fails. STUDY DESIGN: From January 1985 to December 1999, 32 women with cervical pregnancy were treated with methotrexate intramuscularly according to a repeated intramuscular injection protocol. For evaluation of the efficacy of therapy, pretreatment serum concentrations of human chorionic gonadotropin, size of the gestational mass, fetal cardiac activity and presence of fluid in the peritoneal cavity were measured. These findings were analyzed and compared in terms of success and failure by means of the chi 2 test, Fisher's exact test, receiver operating characteristic curve and Student's t test. RESULTS: There was no relation between the women's age, parity, size of the conceptus or presence of fluid in the peritoneal cavity to the efficacy of treatment. In a cervical pregnancy that presented with a serum human chorionic gonadotropin concentration of > or = 10,000 mIU/mL, fetal cardiac activity was associated with a higher failure rate of primary methotrexate treatment. CONCLUSION: In cervical pregnancies, a high serum human chorionic gonadotropin concentration and fetal cardiac activity were the most important factors associated with failure of treatment using methotrexate. PMID- 12092019 TI - Round-headed spermatozoa in semen specimens from fertile and subfertile men. AB - OBJECTIVE: To investigate the frequency of round-headed, or acrosomeless, spermatozoa, determine the percentage and evaluate the possible correlation with other semen parameters. STUDY DESIGN: Semen specimens from 114 subfertile men aged 24-53 years (mean +/- SD 33.3 +/- 6.3) and from 60 fertile men aged 24-44 years (33.1 +/- 4.2) were studied. Two semen specimens were examined from each individual, with a six- to eight-week interval. Sperm morphology was evaluated from Papanicolaou-stained smears, and the classification of abnormal sperm forms was made according to WHO guidelines. RESULTS: The percentage of round-headed spermatozoa was 2.3% +/- 0.5 in subfertile and 0.5% +/- 0.1 in fertile men. Round headed spermatozoa existed in semen specimens from 36.8% of subfertile and 25.0% fertile men. Of subfertile men, 14.9% had round-headed spermatozoa at a higher percentage than the highest normal limit found in sperm smears from fertile men. CONCLUSION: In some subfertile men with a high percentage of round-headed spermatozoa, infertility could be attributed to the cause of this morphologic abnormality. Moreover, morphologic abnormalities in the neck were significantly more frequent in round-headed spermatozoa than in spermatozoa with normal heads. PMID- 12092020 TI - Correlation between plasma neurotransmitters and memory loss in pregnancy. AB - OBJECTIVE: To correlate the levels of plasma neurotransmitters epinephrine, norepinephrine, serotonin and dopamine with memory in healthy, pregnant women. STUDY DESIGN: Fifty healthy, pregnant women were selected in the first trimester and followed in the second and third trimesters of pregnancy. Nonpregnant women served as controls. Epinephrine, norepinephrine, serotonin and dopamine levels were analyzed with high-performance liquid chromatography. The plasma neurotransmitter levels were correlated with memory in each trimester of pregnancy. RESULTS: Significant decreases (P < .001) in plasma epinephrine, serotonin and dopamine were observed in healthy, pregnant women in each trimester of pregnancy when compared to nonpregnant women. A significant increase in plasma norepinephrine was observed in healthy, pregnant women in each trimester of pregnancy. A significant decrease (P < .001) in functional memory was observed in healthy, pregnant women when compared to nonpregnant women. CONCLUSION: Decreases in functional memory and of plasma epinephrine, norepinephrine and serotonin levels in the second trimester of healthy pregnancy suggests that decreased plasma neurotransmitter levels are responsible for loss of functional memory in healthy, pregnant women. PMID- 12092021 TI - Perinatal outcome in large-for-gestational-age infants. Is it influenced by gestational impaired glucose tolerance? AB - OBJECTIVE: To examine the relationship between the World Health Organization category of impaired glucose tolerance (IGT) (two-hour value of the 75-g oral glucose tolerance test at 8-10.9 mmol/L) and outcome in large-for-gestational age (LGA) infants to determine whether IGT affects perinatal morbidity in addition to affecting infant size. STUDY DESIGN: A retrospective study was performed on 461 LGA newborns (birth weight > 90th percentile) from singleton pregnancies delivering after 36 completed weeks in a 12-month period to determine the difference in perinatal outcome between nondiabetic pregnancies (n = 382) and pregnancies with diet-treated IGT (n = 79). RESULTS: The IGT group had significantly higher mean maternal age, prepregnancy weight and body mass index (BMI) but lower absolute and percent gestational weight gain and no difference in infant gestational age, birth weight, BMI, incidence of macrosomia (birth weight > or = 4,000 g) or obstetric complications. However, the IGT group had an increased incidence of Erb's palsy (OR 7.81, 95% CI 1.76-34.62), meconium aspiration syndrome (OR 5.29, 95% CI 1.27-22.02), phototherapy (OR 2.10, 95% CI 1.03-5.69), sepsis (OR 2.90, 95% CI 1.25-6.74) and shoulder dystocia (OR 5.64, 95% CI 1.06-29.89) after adjusting for confounding factors (maternal age and BMI, postdate pregnancy, mode of delivery and infant sex). CONCLUSION: Despite dietary treatment, maternal IGT is associated with increased perinatal morbidity independent of its effect on fetal size. PMID- 12092022 TI - Twin gestation two years after the diagnosis of premature ovarian failure in a woman on hormone replacement therapy. A case report. AB - BACKGROUND: Premature ovarian failure (POF) is a condition causing amenorrhea, hypoestrogenism and elevated gonadotropins before the age of 40 years and affects around 10% of patients seeking evaluation for secondary amenorrhea. Although it is a rare event, pregnancy has occurred in this group of patients. CASE: A successful twin pregnancy occurred in a 30-year-old woman two years after the diagnosis of POF. Cyclic hormone replacement therapy was initiated after the diagnosis. Twenty-one months later, ultrasound revealed an 8-week twin pregnancy. Two healthy infants, a boy and a girl, were delivered at 36 gestational weeks. CONCLUSION: There are no clinical or laboratory features that unequivocally establish the diagnosis of POF. Neither high levels of gonadotropins nor an ovarian biopsy with rare follicles excludes the possibility of pregnancy. Even though pregnancy in patients with POF is very unlikely, patients should be informed of this possibility, and a contraceptive method should be considered if pregnancy is not desired. PMID- 12092023 TI - Allergic contact dermatitis from transdermal estradiol and systemic contact dermatitis from oral estradiol. A case report. AB - BACKGROUND: About 20% of patients using transdermal estradiol complain of adverse local side effects. CASE: A 47-year-old, postmenopausal woman developed eczematous lesions at the sites of application of a estradiol therapeutic transdermal system and successively at the sites of application of a gel containing estradiol. Due to the topical intolerance, the therapy was switched to oral estrogen, which caused a systemic pruritic rash. Positive patch tests with estradiol led to the diagnosis of type IV allergic dermatitis due to transdermal estradiol and to a gel containing estradiol. Systemic contact dermatitis due to oral estradiol was also diagnosed. CONCLUSION: Even though allergic contact dermatitis from estradiol is extremely rare, local side effects from estradiol systems must be kept in mind and correctly diagnosed. Patch tests allow identification of the causative agent. In the case of primary sensitization to topical estradiol, oral estrogens must be prescribed cautiously to avoid systemic reactions. PMID- 12092025 TI - FISH analysis of Y chromosome long arm deletions in subfertile men considering ICSI. A report of two cases. AB - BACKGROUND: Men with Y chromosome long arm deletions, resulting in infertility, form a very significant group of patients, with a view to treatment. With recent advances in assisted reproductive techniques, it is expected that if these patients undergo intracytoplasmic sperm injection, their male offspring will inherit the same deleted regions of Y chromosomes. Hence, characterization of these deleted Y regions provides information, allowing patients to make informed decisions about reproduction. Fluorescence in situ hybridization (FISH), with probes along the Y chromosome, is very helpful in identification of the exact breakpoints. CASES: Two men with complaints of infertility on examination showed low or no sperm in their semen samples. Routine cytogenetic analysis of the peripheral blood showed a small marker chromosome. This marker was identified as the Y chromosome by FISH. Subsequently, other probes along the Y long arm were used to characterize the extent of the deletion. CONCLUSION: With the use of fluorescence-labelled probes, it was possible to identify the extent of the Y chromosome deletion. The distal Yq11 region had been lost in both patients, resulting in oligospermia and azoospermia. Characterizing markers by FISH gave good guidelines to the patients about the possible effects on offspring, allowing them to make appropriate decisions. PMID- 12092024 TI - Concurrent appendicitis and ectopic pregnancy. A case report. AB - BACKGROUND: Ectopic pregnancy is infrequently encountered together with appendicitis. Since 1960, 21 such cases have been reported. CASE: A 32-year-old, Hispanic woman presented with signs and symptoms of an acute surgical abdomen. Preoperatively, the diagnoses of both ectopic pregnancy and acute appendicitis were entertained. On surgical exploration, a ruptured appendix and an unruptured left-sided ectopic pregnancy were found. CONCLUSION: The possibility of multiple pathologic disorders should always be considered in a patient with an acute surgical abdomen, especially during pregnancy. Evidence exists of a possible association between ectopic pregnancy and appendicitis in terms of a common pathogenic mechanism. PMID- 12092026 TI - Immunologic aspects of tropical lung disease. AB - Respiratory tract infections are a major cause of morbidity and mortality in adults and children worldwide. Because of its anatomical features, which allow gaseous exchange, the respiratory tract is constantly exposed to the outer environment and to the systemic and pulmonary circulation, which may allow infectious microbes, toxins, allergens, dust, and other antigens to enter the lung. The human host is a perpetual battleground between the body's immune system and invading antigens, whether they are microorganisms, chemicals, or cancer cells. Although a vast amount of literature is accumulating on the subject of immune responses to pathogens, the mechanisms underlying specific immunity to many organisms remain unknown. Paradoxically, while the immune response has evolved to confer protection against invading antigens, much human pathology arises when the immune responses are evoked. PMID- 12092027 TI - Lung radiology in the tropics. AB - A high-quality chest radiograph and a timely, accurate report are often impossible in the tropics. Rationale matching of service to need, enthusiasm, commitment, and exploitation of information technology all go some way toward enabling patients with pulmonary disease to be imaged. The radiologic findings reflect the high preponderance of infectious disease. TB, HIV, and TB modified by HIV may be "routine" features in some parts of the tropics. Elsewhere, infestation with ameba, hydatid, and strongyloidosis, paragonimiasis, and melioidosis accounts for radiographic signs. The key is to have these conditions firmly in mind when reading tropical radiographs and be aware that the pattern of disease may be different between the patient from the tropics and the more familiar patient from downtown New York. PMID- 12092029 TI - Tuberculosis in the tropics. AB - Tropical countries bear the brunt of the global TB burden. Young children are at high risk and suffer the most severe forms of TB; adults with pulmonary cavities are the main sources of transmission. The incidence in sub-Saharan Africa is increasing as a consequence of the HIV pandemic. Smear-negative TB, which is common in children and patients who have HIV infection, is becoming a major problem in resource-poor settings where access to mycobacterial culture and histopathology is limited. Clinical case definitions are being developed to address this problem. Short courses of rifampin-based therapy are not universally available, but access is increasing. DOTS is the main strategy that the WHO is promoting to improve TB control. This is particularly important for sputum smear positive patients. Unfortunately, the DOTS targets set by the WHO have not yet been met. Innovative, low-cost ways of supervising therapy have been developed using family members or lay supervisors. Preventive therapy in tropical countries is limited to high-risk cases (young children and HIV-infected patients who are tuberculin skin test-positive). An improved TB vaccine would dramatically improve TB control. PMID- 12092028 TI - Granulomatous lung diseases in the tropics. AB - International travel, changing immigration patterns, and political upheavals have transformed the epidemiological picture of many tropical granulomatous disorders [53]. These diseases are seen with an increasing frequency in the United States, Europe, and other developed countries. Physicians must now become familiar with illnesses that were previously regarded as being "exotic" because of this increasing prevalence. Accurate diagnosis is important because therapy is significantly different for infectious granulomatous disorders than for non infectious diseases. PMID- 12092030 TI - Extrapulmonary tuberculosis in high prevalence of tuberculosis and low prevalence of HIV. AB - The authors' data show a higher rate of pleural and meningeal involvement among extrapulmonary TB cases than expected by previous works. Special attention should be given to tuberculous meningitis cases among all extrapulmonary TB cases because of its high mortality rate. The most common extrapulmonary involvement is pleural. Pleural involvement is most common among the young male military service personnel. These data underscore the importance of determining pleural involvement among extrapulmonary TB cases and emphasize the need to consider clinic and epidemiologic differences in the diagnosis and evaluation of extrapulmonary TB. Finally, it seems unlikely that HIV infection currently has a role in the cause of extrapulmonary TB in the authors' region. PMID- 12092031 TI - Pulmonary manifestations of HIV/AIDS in the tropics. AB - The most significant pulmonary opportunistic infections in the tropics are TB and pneumococcal pneumonia. Guidelines for the diagnosis and management of these and other pulmonary manifestations of HIV are discussed. Ultimately, unless concerted efforts are made to treat underlying HIV infection in regions most devastated by AIDS, the impact of these diseases will continue to grow. PMID- 12092032 TI - Non-tuberculous mycobacterial pulmonary infections. AB - Pulmonary disease due to EM occurs worldwide, and its prevalence has increased as a consequence of the HIV pandemic. It is not often detected in the tropics owing to a lack of laboratory facilities, but when sought it has been found. In addition to HIV infection certain occupations such as mining render the work force more susceptible to disease and calls for a revision of working conditions. Resolution by therapy can be achieved in many cases. As the prevalence of TB diminishes worldwide--and hopefully it will in the wake of the resurgence of interest and the widespread application of the World Health Organization's Directly Observed Therapy Short Course (DOTS) strategy--disease due to EM will become relatively more important and will necessitate revised strategies in clinical, microbiological, and public health approaches to mycobacterial disease. PMID- 12092033 TI - Eosinophilic lung disease in the tropics. AB - Paul Ehrlich first described the eosinophil in 1879. It is a pro-inflammatory cell that is involved in the immune defense against various offenders in atopic and nonatopic individuals. Eosinophils derive from an unknown progenitor cell in the bone marrow. This article reviews the etiology and effects of this condition. PMID- 12092034 TI - Hydatid lung disease (echinococcosis/hydatidosis). AB - Two Echinococcus species may exhibit medical relevance as causative agents of pulmonary forms of echinococcosis. Most importantly, infections with Echinococcus granulosus result in "cystic hydatid disease" or "cystic echinococcosis," which affects the lungs in a considerable ratio of cases. Echinococcus multilocularis, which causes "alveolar echinococcosis," affects the lungs relatively rarely and then usually upon metastasizing from primary hepatic lesions. Cystic echinococcus and alveolar echinococcus differ pathologically and clinically so greatly that they are considered separately in this article, although alveolar echinococcus is covered minimally because of its minor importance regarding lung infections. PMID- 12092035 TI - Paragonimiasis: a Japanese perspective. AB - Paragonimiasis has been considered to be a foodborne zoonosis endemic only in limited areas in the world. Recently, however, patients have been seen almost all over the world because of the increase in number of overseas travelers and the popularization of ethnic dishes in developed countries. If paragonimiasis is misdiagnosed as tuberculosis or lung cancer patients suffer from a considerable burden of long-term hospitalization and unnecessary examinations and treatments. Clinicians should always be aware of the possibility of paragonimiasis when patients have pulmonary lesions with eosinophilia and an elevated serum IgE. For the diagnosis, rapid and reliable immunodiagnostic methods are now available. Highly effective drugs are also available for treatment. PMID- 12092037 TI - Pulmonary schistosomiasis. AB - Schistosoma infection is one of the most common infectious diseases, limited in the past only to the endemic countries. With the enormous increase in migration and travel, we encounter more and more cases in developed, nonendemic countries. Although the disease has been known for many years from studies in the endemic countries, the new patient population of nonimmune travelers presents with a different clinical pattern that requires further investigation. One of the features of the disease in the nonendemic population is pulmonary involvement that seems to be much more common than previously suspected. The differences between the nonimmune population with the early pulmonary involvement and the population of endemic areas with late pulmonary involvement are summarized in Table 1. Clinicians in the Western countries have a higher chance of encountering the early (acute) form of the disease, although immigrants from endemic countries may present with late (chronic) schistosomiasis. In the differential diagnosis of pulmonary pathology, especially when accompanied by eosinophilia, schistosomal infection should be considered. The travel history of the patient is mandatory for an evaluation. PMID- 12092036 TI - Paragonimiasis: a view from Columbia. AB - Paragonimiasis is a zoonosis caused by adult trematodes of the Paragonimus genus. The infection in humans is a result of a complex transmission cycle that includes two obligate intermediate hosts, a snail and a crustacean or a crayfish, and a definitive mammalian host. It has been shown that 9 of the more than 40 species of Paragonimus described affect humans in over 39 countries in Asia, Africa and America. It is estimated that 20.7 million people have paragonimiasis and it is calculated that 195 million people are at risk of being infected. The illness usually is caused once the parasite has settled in the lung at the site of the main clinical symptoms: cough, thoracic pain and hemoptysis. The diagnosis of paragonimiasis is based on the patient's history, the parasitological findings (ova in sputum and in feces), and the result of radiological and immunological tests. In severe cases, the patient may suffer from life-threatening hemoptysis or pneumothorax. Currently, praziquantel is the drug of choice. PMID- 12092038 TI - Adult respiratory distress syndrome in the tropics. AB - Today ARDS is more frequently recognized and managed in tropical countries, although published data from most locations is meager. The spectrum of disorders causing ARDS in tropical countries includes virtually all conditions encountered in the West. Additionally, tropical infections and other disorders are seen far more commonly. In particular, malaria and TB are important infections that predispose patients to ARDS in the tropics. Both of these illnesses give lead to severe forms of disease, such as falciparum malaria, acute miliary TB or TB bronchopneumonia, and may cause ARDS. Awareness of the complications helps in early recognition and differential diagnosis from several similar manifestations. Although earlier reports painted a gloomy picture of the outcome of these patients in general--mainly due to financial and logistic constraints--the scenario is improving quickly with better and wider availability of newer diagnostic and management tools. PMID- 12092039 TI - Malaria and the lung. AB - Pulmonary edema that results from increased pulmonary capillary permeability is the most important pulmonary manifestation of malaria. It is a common feature of severe malaria but also occurs rarely in milder disease. Mortality rate is high. The pathophysiologic basis is unclear. In the field, there is much clinical overlap between malaria and pneumonia in children. For physicians in nonmalarial areas, malaria always should be considered in the differential diagnosis of a sick patient who has traveled to a malaria-endemic area. More research is needed to better define and tailor treatments for malarial and nonmalarial ALI and ARDS. PMID- 12092040 TI - Pulmonary complications of leptospirosis. AB - Leptospirosis is a worldwide disease. In this time of globalization knowledge about leptospirosis is important. Although pulmonary involvement has an incidence varying from 20% to 70% and its exteriorization may vary from mild to severe, The severe form appears to be becoming more prevalent (at least in Brazil) and may be associated with higher mortality. PMID- 12092041 TI - Thoracic amebiasis. AB - Pleuropulmonary amebiasis is the common and pericardial amebiasis the rare form of thoracic amebiasis. Low socioeconomic conditions, malnutrition, chronic alcoholism, and ASD with left to right shunt are contributing factors to the development of pulmonary amebiasis. Although no age is exempt, it commonly occurs in patients aged 20 to 40 years, with an adult male to female ratio of 10:1. Children rarely develop thoracic amebiasis: when it does occur there is an equal sex distribution. The infection usually spreads to the lungs by extension of an amebic liver abscess. Infection may pass to the thorax directly from the primary intestinal lesion through hematogenous spread, however. Lymphatic spread is one possible route. Inhalation of dust containing cysts and aspiration of cysts or trophozoites of E histolytica in the lungs are some other hypothetical routes. The lung is the second most common extraintestinal site of amebic involvement after the liver. Usually the lower lobe, and sometimes the middle lobe of the right lung, are affected, but it may affect any lobe of the lungs. The patient develops fever and right upper quadrant pain that is referred to the tip of the right shoulder or in between the scapula. Hemophtysis is common. The diagnosis of thoracic amebiasis is suggested by the combination of an elevated hemidiaphragm (usually right), hepatomegaly, pleural effusion, and involvement of the right lung base in the form of haziness and obliteration of costophrenic and costodiaphragmatic angles. Infection is usually extended to the thorax by perforation of a hepatic abscess through the diaphragm and across an obliterated pleural space, producing pulmonary consolidation, abscesses, or broncho-hepatic fistula. Empyema develops when a liver abscess ruptures into the pleural space. Rarely, a posterior amebic liver abscess can burst into the inferior vena cava and develop an embolism of the inferior vena cava and thromboembolic disease of the lungs with congestive cardiac failure or corpulmonale. Diagnosis by finding E histolytica in stool specimens is of limited value. In a limited number of cases amebae might be found in aspirated pus or expectorated sputum. "Anchovy sauce like" pus or sputum may be found. Presence of bile in sputum indicates that the pus is of liver origin. Serological tests are of immense value in diagnosis. Liver enzymes are usually normal and neutrophilic leucocytosis may or may not be found. ESR is invariably elevated. Anti-amebic antibodies can be detected by ELISA, IFAT, and IHA. Amebic antigen can be detected from serum and pus by ELISA. Detection of Entamoeba DNA in pus or sputum may be a sensitive and specific method. Pleuropulmonary amebiasis is easily confused with other illnesses and is treated as pulmonary TB, bacterial lung abscesses, and carcinoma of the lung. A single drug regimen with metronidazole with supportive therapy usually cures patients without residual anomalies. Aspiration of pus from empyema thoracis may be needed for confirmation and therapeutic purposes. The pericardium is usually involved by direct extension from the amebic abscess of the left lobe of the liver, sometimes from the right lobe of the liver, and rarely from the lungs or pleura. An initial accumulation of serous fluid due to reactive pericarditis followed by intrapericardial rupture may develop either (1) acute onset of severe symptoms with chest pain, dyspnea, and cardiac tamponade, shock, and death, or (2) progressive effusion with thoracic cage pain, progressive dyspnea, and fever. Chest radiograph, ultrasound examination, and CT scan usually confirm the presence of a liver abscess in continuity with the pericardium and fluid within the pericardial sac with or without the fistulous tract. Echocardiography may demonstrate fluid in the pericardial cavity. Patients should be cared for in the ICU and ambecides should be started without delay. Pericardiocentesis usually confirms the diagnosis and improves the general condition of the patient. Aspiration of the accumulated fluid should be performed urgently in cardiac tamponade; repeated aspiration may be needed. Surgical drainage should be done if needed. Acanthamoeba, a free-living ameba, may also infect the lungs in the form of pulmonary nodular infiltration and pulmonary edema in association with amebic meningoencephalitis in immunocompromised patients. It usually spreads to the meninges of the brain by way of the blood from its primary lesion in the lung or skin. Early diagnosis and institution of treatment may be life saving for these patients. A literature review shows that HIV/AIDS patients are not prone to infection with E histolytica. It is now clear that there are an increasing number of HIV-seropositive patients among amebic liver abscess patients, however, which suggests that although the incidence of intestinal infection is not high among HIV-seropositive or AIDS patients they are more susceptible to an invasive form of the disease. PMID- 12092042 TI - Pulmonary complications of Behcet's disease. AB - Our knowledge about pulmonary complications of Behcet's disease continues to evolve, but we need controlled trials for the management of the disease. The main goal should be to elucidate the pathogenesis and standardize the management according to the underlying pathologic process. PMID- 12092043 TI - Thoracic and lung involvement in familial Mediterranean fever (FMF). AB - Lung involvement in FMF is limited mainly to transient pleuritis during acute attacks. Amyloidosis of the lung is rare and is associated with symptomatic involvement of other organs while remaining subclinical in itself. Vasculitis of the lung in FMF is possible because of the strong association between FMF and a variety of vasculitides. With the exception of one case of isolated pulmonary vasculitis, vasculitis of the lung in FMF has not been described. The claim that FMF protects against asthma has not been established, but this inverse association, if present, may be traced to linkage disequilibrium in which MEFV modifies the effect of asthma and atopic-related genes, or to eosinophil function. Mesothelioma has been reported in at least four patients with FMF and is related to chronic or recurrent stimulation of the serous membrane. Three patients had peritoneal mesothelioma, while one developed mesothelioma of the lung. Finally, thromboembolism should be considered, particularly in patients with FMF amyloidosis who present with respiratory distress. PMID- 12092044 TI - Whipple's disease: a granulomatous masquerader. AB - Whipple's disease is a multisystemic infection that affects middle-aged white men. It typically presents with fever, polyarthritis, diarrhea, steatorrhea, and weight loss. Many other systems can be involved, however, including the central nervous system, heart, lymphatics, lungs, bone marrow, and skin. Recent work has demonstrated the causative organism to be a complex bacteria, Tropheryma whipplei. The diagnosis is established most securely by periodic acid-Schiff staining of foamy monocyte-macrophages in biopsy tissue and body fluids, by electron microscopy, which reveals bacilli within membrane-bound vesicles, and by molecular amplification techniques using polymerase chain reaction of tissues and body fluids. The differential diagnosis includes chronic multisystemic infections and granulomatous disorders, because Whipple's disease is a fascinating blend of both. The condition can resemble sarcoidosis and mycobacterial disease and fungal, protozoal, and bacillary infections. Earlier diagnosis leads to earlier treatment and hopefully the prevention of chronic disabling complications and needless mortality from this once uniformly fatal condition. PMID- 12092045 TI - Control of Chagas disease. AB - Chagas disease occurs throughout Mexico and central and southern America and continues to pose a serious threat to health in many countries of the region. People infected with the trypanosome parasite may suffer cardiac, gastrointestinal, or neurological damage, although disease manifestations vary widely from one endemic area to another. In the past decade, control programmes in several endemic countries have proved remarkably successful; interruption of disease transmission has been achieved in some and is imminent in others, leading to a substantial reduction in the incidence of Chagas disease in Latin America. However, 8-9 million people in Mexico and the Andean and central American countries are infected with the parasite and 25 million remain at risk, emphasizing the need to sustain and extend control strategies. This report of a WHO Expert Committee reviews current knowledge of Chagas disease and its pathogenesis, discusses the causative parasite, the triatomine vectors, and the natural reservoirs of infection, and considers the epidemiology and incidence trends of the disease. Prevention and control strategies are described, as are the various formal initiatives for interruption of disease transmission. The report concludes by identifying priorities for research and offering guidance for the planning, implementation, and strengthening of national control programmes. PMID- 12092046 TI - [The failing heart is helped by a mechanical pump. Prolonged survival and better quality of life in the first randomized trial]. PMID- 12092047 TI - [A protective network for catching misleading laboratory results is required]. PMID- 12092048 TI - [Great number of non-recorded cases of analytical interference. Unexpected and misinterpreted laboratory levels can result in diagnostic and therapeutic errors]. PMID- 12092049 TI - [Physicians' well-being is deteriorating. Results from a prospective intervention study. The mental energy decreases as workload increases]. AB - The impact of the current transformation of health care on the well-being and working conditions of physicians has been analyzed in a 6-year prospective intervention study at the Orebro University Hospital (USO). Physicians' ratings of their work, organization and health have been compared with those of other health care professionals at USO, as well as with ratings by physicians in other hospitals. Feedback on survey results has been periodically provided to employees in all hospital departments as well as to hospital senior management. The results have formed the basis for a range of improvement initiatives. The study identified a number of positive changes, such as increased ratings on scales measuring social climate, skills development and participatory management. However, workload has increased and mental energy decreased during the same period. PMID- 12092050 TI - [Risks in physicians' working environment. From plague and anesthesiologic gases to stress and lack of control]. PMID- 12092051 TI - [With due reflection and own responsibility can working conditions improve]. PMID- 12092052 TI - [The brain behind stress and fatigue depression]. PMID- 12092053 TI - [Incidents in health care--punishment or committee of inquiry?]. PMID- 12092054 TI - [New economic control system can affect physicians' working environment]. PMID- 12092055 TI - [Working environment within health care services--from words to action]. PMID- 12092056 TI - [Why is the Sound bridge a successful example of prevention of occupational dermatological disease?]. PMID- 12092057 TI - [The role of physicians in the increasing sick leave statistics--what do we want and can do?]. PMID- 12092058 TI - [HPV testing--the effect must be evaluated prior to clinical use!]. PMID- 12092059 TI - [Don't administer NSAID after bone surgery!]. PMID- 12092060 TI - [Why change the order of preference between NSAID and mild opioids in ambulatory care?]. PMID- 12092061 TI - [A comment concerning the proposal for a competence-center for better hearing]. PMID- 12092062 TI - [Measurement of cerebral flow velocity--some viewpoints]. PMID- 12092063 TI - [Physicians as branded goods]. PMID- 12092064 TI - [Post-modern anti-vaccination movement]. PMID- 12092065 TI - [Cooperation between competitors?]. PMID- 12092066 TI - [Licorice--not just candy]. AB - BACKGROUND: Liquorice is widely used as a flavour and also as a medical drug. Possible side effects include hypertension, hypokalaemia and metabolic alkalosis. MATERIALS AND METHODS: We present a case history and a review on liquorice intoxication based on relevant literature found from searches on Medline. RESULTS AND INTERPRETATION: A 19-year-old girl was admitted to hospital with severe hypertension, hypokalaemia and metabolic alkalosis. Urine analysis showed inhibition of 11-beta-hydroxysteroid dehydrogenase. It turned out that she ingested a lot of liquorice; after she stopped eating it she became normotensive without any medication. Three months later there were no signs of inhibition of the enzyme. The active component of liquorice is glycyrrhetinic acid, which inhibits the enzyme 11-beta-hydroxysteroid dehydrogenase. This enzyme promotes the conversion of cortisol to cortisone and is thereby responsible for the specificity of the mineralocorticoid receptor to aldosterone in the collecting tubules. Inhibition of the enzyme allows cortisol to act as the major endogenous mineralocorticoid producing a marked elevation in mineralocorticoid activity, resulting in hypertension, hypokalaemia and metabolic alkalosis. PMID- 12092068 TI - [Quality of life, gender, complaints and humor among dialysis patients]. AB - BACKGROUND: This study tested the correlation between quality of life and sense of humour as well as the sociodemographic and biomedical characteristics of patients in dialysis due to renal failure. Gender differences in all these variables were also tested. MATERIAL AND METHODS: All patients in need of dialysis in Sor-Trondelag county, Norway in February 1998 (N = 52) were recruited; 46 patients completed the survey (32 males and 14 females). RESULTS: Males were more likely to be married and have a house of their own than were women. Correlation analysis stated that high quality of life was significantly associated with low comorbidity and a good sense of humour. The most efficient model to explain quality of life in multiple regression included serum albumin (not significant) and confirmed the independent contributions from low comorbidity and a good sense of humour. INTERPRETATION: Comorbidity and a sense of humour were factors of greater importance than serum albumin to differences in quality of life among patients on dialysis. PMID- 12092067 TI - [Serious mushroom poisoning by Cortinarius and Amanita virosa]. AB - BACKGROUND: Following a characteristic long latent period (3-17 days), the nephrotoxins of Cortinarius rubellus and Cortinarius orellanus can cause the orellanus syndrome, due to severe damage of the proximal tubular epithelium. Amanita virosa is known to produce serious toxic effects in the liver and the kidneys after an initial asymptomatic latent phase. MATERIAL AND METHODS: We discuss the toxicity, clinical features and treatment of the orellanus and the phalloides syndromes and present six case histories. RESULTS AND INTERPRETATION: Ingestion of Cortinarius rubellus and Cortinarius orellanus resulted in permanent renal failure in four out of five patients, following a latent period of about ten days. One patient who ingested Amanita virosa, developed hepatotoxicity. He was given silibinin and symptomatic treatment and recovered. After ingestion of Cortinarius rubellus and Cortinarius orellanus, no specific treatment is available. The therapy is directed toward the renal failure, including dialysis and possible transplantation. Poisoning by Amanita virosa is treated with the nonspecific antidote silibinin. PMID- 12092069 TI - [Disease anxiety among medical students and law students]. AB - BACKGROUND: Medical students and other people with knowledge about health and disease are often thought to have more anxiety about their health than others. MATERIAL AND METHODS: 80 medical students and 100 law students were asked to fill in a questionnaire designed to identify higher disease anxiety (Whiteley Index). A high total score on the Whiteley Index indicates high disease anxiety. 62 medical students (78%) and 70 law students (70%) returned the questionnaire in an anonymous form. RESULTS: Medical students had lower total score compared with law students (p < 0.01). One medical student and six law students had a total score susceptible of hypochondriasis. Male students had a non-significantly higher mean total score than female students. Those with depressive mood (n = 5), had higher total score than the others (p < 0.01.) Those who had experienced disease in their family (n = 83) tended to have lower total score. INTERPRETATION: Medical students have less disease anxiety than law students, contrary to what is often believed. PMID- 12092070 TI - [The brothers of Jumiege--the peripheral nervous system in early French mythology]. AB - This article reviews the process of discovery of the nervous system from Pythagoras (570-500 BC) to Galen (130-201 AD). After Galen, no anatomical studies were performed before the renaissance. According to a legend, probably produced for political reasons, two brothers, sons of the French king Clovis II, revolted against their father and were sentenced to loose their physical powers by having the nerves of their arms and legs cut. They were then set adrift on the river Seine, stranding at the Jumiege monastery. The earliest written version of this legend stems from the fourteenth century; it was probably a part of the local French mythology. This indicates that the existence of the peripheral nervous system, and therefore also in part the knowledge contained in the early anatomical works, quite early may have been more or less known outside academic circles. PMID- 12092072 TI - [Working environment and job adjustment among bus drivers]. AB - BACKGROUND: Bus drivers as a group have elevated health risks that are related to their working conditions. The present study measured the perception of working conditions among workers in a bus company and the results of job adjustments for health problems. MATERIAL AND METHODS: Perception of working conditions was measured by a questionnaire in a cross-sectional design in which bus drivers were compared to workers in other departments of the same company. In a longitudinal design, effects of intervention were studied when a small group of workers experiencing health problems were offered job adjustments for shorter or longer periods. RESULTS: Duration of employment was associated with a negative perception of working conditions, particularly among bus drivers with more than 10 years' experience. Between one and three years after intervention (job adjustment), 80% of the workers recruited to the intervention were still in their jobs; 60% of the workers needed only a mean period of six months of job adjustment. INTERPRETATION: A wear-and-tear effect could be associated with number of years of driving. Preventive measures to reduce further health problems among these drivers could involve job adjustments. PMID- 12092071 TI - [The skin as an immunological organ]. AB - BACKGROUND: The skin is our largest organ. Its physico-chemical properties are adapted to its function as a non-specific general defense line against infections and injury from the environment. Through evolution the skin has developed a second line of highly specific immune defense comprising epidermal Langerhans cells, T lymphocytes and keratinocytes as the most distinct cellular elements. MATERIAL AND METHODS: On the basis of available research data and reviews, an overview on the skin as an immunological organ is presented. RESULTS AND INTERPRETATION: Antigens penetrating into the epidermis are bound to dendritic Langerhans cells, processed and presented to T lymphocytes. Certain memory T cells are able to remember where they first encountered antigen. Skin homing T cells are CLA+ and circulate between the skin, peripheral blood and skin draining lymph nodes. Keratinocytes produce a wide spectre of cytokines, in particular interleukin-1, which has a key role in the skin immune function. The most common inflammatory skin disorders are mediated by T lymphocytes, usually Th1 cells, as in psoriasis, or Th2 cells, as in atopic dermatitis. Systemic treatment with T cell specific suppressive drugs, such as cyclosporine, has been successful in certain dermatoses. Promising cyclosporine analogues for topical treatment (tacrolimus and pimecrolimus) have been developed and may soon be introduced. PMID- 12092073 TI - [Interdisciplinary working teams--better for both the patients and the staff]. AB - BACKGROUND: Patients in Norwegian hospitals often feel that they do not know which specialist is responsible for their treatment. We report on a reorganisation carried out in the orthopaedic department at Buskerud county hospital in 1997-98. MATERIAL AND METHODS: Staff members are allocated to five groups, each responsible for 12-15 beds and including one or two specialists, one or two residents, nurses, physiotherapists and secretaries. Patients are treated by the same group throughout their stay in hospital; the group's specialist is responsible for each patient's treatment. RESULTS: A study established that patient satisfaction with the organisation of the department was higher in 1998 and 2000 than in 1996, and more patients felt that one specialist was responsible for their treatment. A high percentage of staff members were satisfied with the reorganisation. The system may, however, be vulnerable, as it demands exact planning and a high degree of staff loyalty. INTERPRETATION: The results indicate that this mode of organisation benefits patients as well as staff. PMID- 12092074 TI - [When anesthesia came to Norway]. AB - On 16 October 1846, William T.G. Morton performed the first successful demonstration of ether anaesthesia. The news quickly spread to Europe; reports reached the Scandinavian countries in early 1847. The first ether anaesthesia was administered in Norway at Rikshospitalet, the teaching hospital of the only Norwegian university at the time, on 4 March 1847. The number of surgical procedures increased; medical practice was changed. PMID- 12092075 TI - [Delirium in old age can be prevented and treated]. AB - BACKGROUND: Delirium is a common neuropsychiatric syndrome characterized by disturbance of attention and consciousness developing over a short period of time. Symptoms tend to fluctuate during the course of the day. Delirium is by definition a direct physiological consequence of a general medical condition and is probably the most common presenting symptom of disease in old age. MATERIAL AND METHODS: Literature review based on search on PubMed and Medline up to 31 December 2001 and a summary of several doctoral theses and our own clinical experience. RESULTS AND INTERPRETATION: Several recent intervention studies have clearly shown that delirium can be prevented and treated. Successful intervention programmes have been multifactorial and interdisciplinary, including assessment and treatment of underlying causes as well as prevention and treatment of factors endangering cerebral metabolism. In particular, cerebral hypoxemia caused by i.e. sleep-apnoea syndrome, anaemia, hypotension, pulmonary diseases, and heart failure is often easily prevented and treated. Excellent nursing care seems to be a prerequisite for successful prevention and treatment of delirium. Acceptable scientific evidence for pharmacological treatment is still lacking; it is sometimes necessary but should be used with caution. If sedation is acceptable, clomethiazole is the drug of choice but if the delirium is complicated by frightening hallucinations and agitation, haloperidol or risperidone can be used but only for short periods. Cholinesterase inhibitors are probably a better choice, though randomised treatment studies are still lacking. PMID- 12092076 TI - [Treatment of malnutrition in elderly patients]. AB - BACKGROUND: Although often undiagnosed, undernutrition is prevalent among elderly patients in hospitals. The effect of undernutrition is reduced mental and pulmonary function, increased prevalence of infection, and increased morbidity and mortality. MATERIAL AND METHODS: This article presents an overview of the diagnosis and treatment of undernutrition in elderly patients with nonmalignant disease, based on relevant literature. RESULTS: There are no generally accepted definitions of undernutrition. Undernutrition should be suspected clinically and screened for, using anthropometric methods (body weight, body mass index, triceps skinfold, arm muscle circumference) or functional tests (hand dynamometry, laboratory parameters). Results from several studies indicate that nutritional intervention could increase body weight and physical capacity of patients and reduce length of hospitalisation and mortality. INTERPRETATION: Undernutrition is common in clinical practice. Hospitalised patients should be screened for risk of undernutrition. The screening should be combined with a nutrition plan, including strategies for nutrition therapy. PMID- 12092077 TI - [Health assessment of the elderly--a practical approach?]. AB - Health assessment of elderly people as a routine procedure has been proposed in official reports and in the literature. Some programmes have been evaluated, including home visits and periodical health assessment programmes, but evaluation produces conflicting results. Whether health assessment of elderly people represents true or false safety depends on the association between risk factors and end points like mortality, morbidity, functional ability and quality of life. Who should be offered a health assessment, what should be the elements, and what is the scientific documentation for such an approach? Health assessment is of little value without follow-up. Periodic large-scale home visits or screening programmes are not recommended. Health assessment of old people at risk should be carried out by the general practitioner. The community has an obligation to plan for adequate medical services to prevent elderly people at risk to deteriorate without medical follow-up. PMID- 12092078 TI - [Nursing home as a medical institution--a fata morgana?]. PMID- 12092079 TI - [Genetic variation--useful information for everybody?]. PMID- 12092080 TI - [The birth and maturation of being a student]. PMID- 12092081 TI - [Guidelines concerning low back pain]. PMID- 12092082 TI - [Was Jesus dead after the crucifixion?]. PMID- 12092083 TI - [Is investing in health economically beneficial?]. PMID- 12092084 TI - [Anthrax and disaster planning at the university hospital in Ulleval]. PMID- 12092085 TI - [Particle pollution--why is the health effect variation so big?]. PMID- 12092086 TI - [Treatment of colonic cancer in the elderly]. PMID- 12092087 TI - [Who is the owner of patient data?]. PMID- 12092088 TI - [A copy of the epicrisis automatically to the patient?]. PMID- 12092089 TI - [The family practitioner system and the English example]. PMID- 12092090 TI - Gout basics. PMID- 12092091 TI - An update on specific COX-2 inhibitors: the COXIBs. PMID- 12092092 TI - Study of human pepsinogen polymorphism by combining chromatographic and electrophoretic methods. AB - A new combination of chromatographic and electrophoretic methods has been developed for better separation and characterization of human pepsinogens. Pepsinogens isolated from the gastric mucosa of patients with gastric cancer have been separated using fast-protein liquid chromatography (FPLC) on an ionex Uno-Q1 column. Proteolytic active fractions were firstly immunodetected by monoclonal antibodies against PGA and PGC using ELISA and then separated by isoelectric focusation in the acidic pH 2.5-5 gradient with an excellent resolution. The combination FPLC and ELISA followed by IEF enabled to separate ten pepsinogen isoforms. This technique is very suitable for studies of the pepsinogen polymorphism and its role in the gastric diseases. PMID- 12092093 TI - [History and present status of surgical treatment of morbid obesity]. AB - Problems of extreme and morbid obesity take on constantly considerable relevance in 21st century. The prevalence of the obesity (BMI--Body Mass Index > 30 kg/m2) is still on the increase worldwide. In the Czech Republic approximately 16% of male and 20% of female in the age of 20-65 years are obese (BMI > 30 kg/m2). To begin with the effective cure is always indicate in case of the obesity over BMI > 30. The bariatric surgery is indicating in the occurrence of failure of conservative care of morbidly obese patients with recurrences of overweight. The morbid obesity with serious associate health complications often represents the vital danger of the patient's life. The development of the bariatric surgery passed from resections of the bowel, gastric bypasses, biliopancreatic diversions, horizontal gastroplasties to in the present the most frequently used methods as the vertical bandage gastroplasty (VBG) and the gastric bandage (GB). The standard applications of the miniinvasive laparoscopic methods with significant decrease of postoperative complications radical changes of the bariatric surgery in the present years. PMID- 12092095 TI - Radiation-induced damage of the grey matter in central nervous system in rats. PMID- 12092094 TI - [Surgical treatment of morbid obesity--gastric banding]. AB - Approximately 16% of male and 20% of female of the age from 20 to 65 years are obese in the Czech Republic. The restrictive bariatric procedure of stomach- gastric banding (GB) is one of possibilities to cure the morbid obese patients after failure of conservative therapy. The ratio of complications (5-18%) after GB presenting in various papers is comparable with the ratio of complications (4 23%) in others bariatric procedures. From 1993 to 1999, 517 morbid obese patients (mean BMI 51.1) underwent laparoscopic nonadjustable gastric banding (LNGB) at 1st Surgical Department, Charles University Teaching Faculty Hospital in Prague. As the early complications (during hospitalization) offered swelling of the gastric mucous in the place of GB in 5.6% (n = 29), the oesophagitis, the gastritis or the gastric ulcer in 1.5% (n = 9) and perforation of the stomach wall in 0.6% (n = 3). As the late complications offered the bleeding from peptic ulcer in 0.4% (n = 2), sequential migration of gastric band through the stomach wall inside in 0.6% (n = 3) and the slippage of anterior stomach wall or the dilatation of the pouch above gastric bandage in 5.1% (n = 26). The serious complications in 6.3% (n = 32) claimed surgical procedures. Other complications in 7.5% (n = 39) have been treated conservatively. The 86% (n = 446) of obese patients after LNGB were without complications. PMID- 12092096 TI - Apoptosis and expression of proliferative proteins in the developing spinal cord and spinal ganglia. PMID- 12092097 TI - Temporal kinetics of calcium in neocortical neuronal nuclei after global cerebral ischemia and reperfusion. An ultrastructural study. AB - New facts about control of nuclear milieu were recently described. Because calcium is known as an important cellular regulator we designed this study to describe temporal calcium kinetics in neuronal nuclei in canine neocortex after ischemia-reperfusion injury. Seven-minute global cerebral ischemia was followed by reperfusion phase lasting from 0 to 24 hours. Ultrastructural histopathology of brain tissue was assessed both qualitatively and semi quantitatively using scoring system. For electron microscopic calcium detection pyroantimonate histochemistry was used. Significant increases in calcium-pyroantimonate deposits were noted in reperfusion time 0, 8 and 24 hours. Reperfusion time 30 minutes showed lower load of calcium deposits than control, this, however, did not reach statistical significance. In conclusion we have found biphasic pattern of nuclear sequestration of calcium in neocortical neurons, which is in agreement with phasic changes of calcium content in other neuronal compartments. PMID- 12092098 TI - Pathomorphological changes in the nodo-paranodal regions of axons after the partial ischemia. PMID- 12092099 TI - Changes in the myelin sheaths after nerve crush. PMID- 12092100 TI - Repetitive ischemia-induced injury in microcirculation of the rabbit spinal cord. PMID- 12092101 TI - Myelinogenesis--longitudinal sections vs serial transverse sections. PMID- 12092102 TI - Morphometry of the peripheral nerve in experimental diabetes mellitus. PMID- 12092103 TI - Contribution to the problem of adrenergic innervation of the kidneys in pheasants and quails. PMID- 12092104 TI - Neuroplacodes and their morphogenetical significance. PMID- 12092105 TI - Detection of apoptosis and regulating proteins Bcl-2 and Bcl-Xs in developing kidney of human embryos II. PMID- 12092106 TI - Ultrastructure of the tracheal epithelium after administration of aminophylline. PMID- 12092107 TI - The role of apoptosis and its regulating genes during differentiation of myocardium of human embroys. PMID- 12092108 TI - The detection of apoptosis by four different immunohistochemical techniques in selected tissues of human embryos. PMID- 12092109 TI - The expression of neuronal acetylcholinesterase (ACHE)-positivity in the human palatine tonsil. PMID- 12092110 TI - Early development of the jugular lymphatics. PMID- 12092111 TI - The effect of a single oral administration of ambroxol on the ultrastructure of the epithelium of terminal bronchioles in rabbits. PMID- 12092113 TI - Creep behaviour of the porcine urinary bladder and its models. PMID- 12092112 TI - The effect of local administration of 2% trimecaine on the glycoconjugate content of goblet cells in the rabbit tracheal epithelium. PMID- 12092114 TI - Mechanical stiffness and morphology of selected human organs from the standpoint of passive safety. Preliminary report. PMID- 12092115 TI - Transposition possibilities of pelvic muscles. AB - A total of 270 muscles, 45 of each of the six individual muscles studied (the sartorius m., the gracilis m., the semitendinous m., the semimembranous m., the greatest gluteal m., the internal obturator m.) were followed up with the aim of evaluating their length parameters and blood and nerve supply. The aim of the study has been to choose the muscle most suitable for transposition and reconstruction of the neosphincter. From the point of view of their anatomy, the internal obturator muscle has been found the most suitable for the purpose. PMID- 12092116 TI - Intraperitoneal and extraperitoneal anastomoses of spleen arteries. AB - The importance of supporting the spleen with blood is enormous especially from the surgical point of view. The main vessel of spleen is the splenic artery. In this study we would like to extend current knowledge of anastomoses between the splenic artery and surrounding arteries. Seven donor bodies of both sexes were used for detailed dissection. We were concerned with both the intraperitoneal and extraperitoneal anastomoses. In another 100 donor bodies, the relationship between the spleen and the anterior surface of the posterior body wall was studied. The superficial anastomoses were found mainly in the peritoneal duplications. They were the short gastric arteries; the epiploic branches of the left gastroepiploic artery; branches of the middle and left colic arteries and arteries from the region of the pancreatic tail. Deep anastomoses were found between the spleen and the parietal branches of the aorta, i.e. the superior and inferior phrenic arteries and the arteries of the anterior surface of the posterior body wall: the posterior intercostal artery and lumbar arteries. Authors also expect small anastomoses in 40-50% of spleens that lie directly on the anterior surface of the posterior body wall. Our findings correspond with the clinical observations that after ligation of the splenic artery necroses in the spleen do not occur. PMID- 12092117 TI - The historical background for the blood supply of the colon. PMID- 12092118 TI - [Viral interactions with the host's immune system]. AB - Viruses are obligatory intracellular parasites, which differ in their structure and strategy of replication. The establishment of an antiviral state in uninfected cells and the elimination of virally infected cells are critical tasks in the host defence. Against the extensive array of immune modalities, viruses have successfully learned how to manipulate host immune control mechanisms. The study of viral strategies of immune evasion can provide insights into host-virus interactions and also illuminates essential functions of the immune system. PMID- 12092119 TI - [Exchange of dentition and its control. Overview of current knowledge and personal experience]. AB - Physiological dentition exchange is often interfered with exogenous and endogenous factors. For this reason it must be steadily regulated. The authors present complex view on this topic based on literature studies and own experiences. The authors define fundamental principles "guided exchange", "tooth eruption" and "eruption path". Correct explication of these ideas is necessary for text understanding. Dentition exchange is considered as complex of eruption interactions of individual permanent teeth including molars. The project is based on idea that dentition exchange time is optimal for preventive interventions. At the end the authors underline the importance of global application of the above mentioned principles on the whole child population. PMID- 12092121 TI - Muscarinic receptor subtypes in the heart--how many and why? AB - Muscarinic receptors are coupled to G proteins and to date, five subtypes have been characterized. These subtypes differ in their effects on second messenger production. Odd-numbered subtypes activate phospholipase C beta, and then they increase the production of inositoltrisphosphate and diacylglycerol. Even numbered subtypes inhibit adenylylcyclase. The expression of individual subtypes is quite tissue specific. The heart tissue is realized as M2 predominant organ (in mammals; the muscarinic population in birds consist of M2 and M4 receptors). This article focuses on the possibility of existence of further muscarinic receptor subtypes in avian and mammalian heart in consequences with activation of phosphoinositid metabolism in the heart. PMID- 12092120 TI - Minimal access surgery in Georgia. PMID- 12092122 TI - ECG body surface isointegral and isoarea maps (BSM) in 30 and 60-years-old healthy humans. AB - ECG body surface isointegral and isoarea maps (BSM) are the sensitive indications of local electrical depolarization and repolarization changes both in controls and in coronary artery disease. In the present work the absolute values of maximum and minimum (extremum) in BSM have been compared in 24 healthy persons (20-36 years) with 18 older ones (54-70) of both sexes, non-smokers and without cardiovascular diseases in their medical history. Twenty-nine parameters of the heart electric field were registered by 96 unipolar electrodes placed regularly on the thorax and analyzed by the system Cardiag. A lower heart rate and a longer QT interval were found in older persons. The maximum (extremum) of isointegral and isoarea maps was less positive and the minimum was less negative in the older than in the younger subjects (p < 0.01). The maximum Q-wave amplitude on surface thorax was significantly lowers in older than in younger persons. The results confirmed the age-dependent decrease of QRS and T wave potentials. PMID- 12092123 TI - [Comparison study of the HUMFGA and HTPO STR system in the Saxon and Czech population]. AB - High variability of repetitive DNA in a non-coding region of human genome affords possibilities for vestige identification and determination of their origin. For analysis of DNA many a time obtained from decomposed material, usually in a very small amount and sometimes partially degraded, polymerase chain reaction is considered as optimal technique. For identification of individuals is used an Amplified Fragment Length Polymorphisms (AmpFLP), which present repetitions 10 to 70 base pairs long in an amplification products of 170 to 1200 bp. In legal medicine PCR systems using tetrameric repetition polymorphism are widely used. These systems are highly sensitive, but in comparison to AmpFLP systems, imperceptibly influenced by nucleic acid degradation in cases of old or partially degraded samples. Aim of our study was to compare frequency data for 2 STR systems, FGA and HTPO, both after PCR amplification, on 200 samples of Czech and 228 of German population. In FGA system of Czech population 14 different alleles and 44 genotypes were determined, while German population provided 13 different alleles and 47 genotypes. Furthermore, frequency of alleles and heterozygosity rate were determined. Heterozygosity rate for Czech samples was 85%, for German ones 87.1%. Alleles 21 and 22 were most frequent (approx. 20%) between both Czech and German population, followed by alleles 20, 23 and 24 (14-12%). Besides these data, three different transient alleles were detected. In HTPO systems of both population groups 12 different genotypes, consisting of 5 alleles, were determined. Genotypes 8/8 and 8/11 observed in 30% were the most frequent ones in both population groups. Heterozygosity rate for Czech samples was 62.5%, for German ones 65%. Results were statistically analyzed. In both population groups, no significant difference from Hardy-Weinberg's balance was found. Additionally, statistic parameters for forensic effectivity evaluation were constructed. Comparison of both populations using homogeneity tests provided non-significant difference. FGA system provides extraordinary high forensic exploitativeness and therefore is fully acceptable for vestige identification and determining of their origin. For significant determination of transient alleles analysis of fluorescein-labelled fragments in denaturing gel system should follow, using automated laser detection of fluorescence. Polymorphism of HTPO-system is significantly lower compared to FGA-system, but relatively small PCR product required as well as simple detection and imperceptible influence of nucleic acid degradation allow the use of HTPO for analysis of highly degraded DNA. PMID- 12092124 TI - [Weight reduction and aspects of the metabolic syndrome]. AB - We carried out analysis of the influence of long-term weight reduction on individual parameters of metabolic syndrome X. We enrolled the total of 30 obese patients (22 with, 8 without syndrome X). During weight reduction, the mean BMI decreased by 4.08 +/- 3.00 kg/m2 leading to significant (p < 0.001) decrease in insulinaemia from 33.4 +/- 25.9 to 21.2 +/- 19.625 mu j/ml. In patients with syndrome X, the decrease in BMI of 3.50 +/- 2.76 kg/m2 was coupled with significant (p < 0.001) decrease in insulinaemia from 39.7 +/- 27.7 to 24.0 +/- 21.725 mu j/ml. Using cluster analysis of cases syndrome X patients formed two distinctive groups with different behaviour. It seems, that diagnosis of metabolic syndrome, based on arbitrary criteria, encompasses stable patients, in our study characterized by higher age and presence of hypertension, and volatile patients, who form somewhat transition stage between simple obesity and fully developed syndrome X. Moreover, relationships between individual parameters at the beginning of the study can elucidate the environmental influences (relationship between insulinaemia and hypertension), whereas those at the end of the study represent true pathogenetic relationships (insulinaemia and glycaemia) and relationships between syndrome X constituents (hypertension, hypertriglyceridaemia and decrease in HDL cholesterol). PMID- 12092126 TI - [Puncture biopsy of peripheral lymph nodes and subcutaneous masses]. AB - Ninety-one consecutive patients with known diagnosis of lung cancer or clinically suspicious malignant tumour underwent fine needle aspiration biopsy of enlarged peripheral lymph nodes or subcutaneous nodular lesions. There were performed 105 fine needle biopsies. Cytological findings of diagnostic value were found out in 56 (53.3%) of cases. Fine needle aspiration biopsy of peripheral lymph nodes and subcutaneous lesions is a simple and valuable method for staging and in some cases for diagnosis of lung tumours. PMID- 12092125 TI - [Sleep disorders in patients treated with continuous ambulatory peritoneal dialysis]. AB - BACKGROUND: A lot of patients with end stage renal disease with the necessity of renal replacement therapy have some sleep problems. The aim of this study was to get basic information about the sleep of patients treated with continuous ambulatory peritoneal dialysis (CAPD), mainly their subjective view on their sleep, including comparison with hemodialyzed patients (HD). METHODS: All patients treated with continuous ambulatory peritoneal dialysis in two dialysis centres were given a simple questionnary containing 20 questions concerning sleep. It was filled in by all these patients--25 patients (mean age 58.1 years)- 12 men and 13 women. Data obtained from 103 hemodialyzed patients from the same two-dialysis centres were used for comparison (mean age 60.4 years)--61 men and 42 women (the same questionnary). RESULTS: 40% of CAPD patients regard their sleep as bad. Thirty six percent of patients have problems with falling asleep, 32% awake three times or more during the night and 28% snore or have some breath problems. CAPD patients feel more frequently tired after the night (32% vs. 18.4% HD), more patients sleep during the day (64% vs. 5.15% HD) and fewer patients feel restlessness of legs (36% vs. 45.6% HD). These differences are not statistically significant. CONCLUSION: Occurrence of sleep disorders and their characteristics in patients on CAPD is similar to that in hemodialyzed patients. PMID- 12092127 TI - [Thrombectomy and embolectomy using the adherent clot catheter]. AB - The efficacy of the Adherent Cloth Catheter is demonstrated on the three above described cases. The different mechanism of function on contrary to the classic Fogarty Catheter enables us to remove the older, strongly adherent thrombus from prothesis, artery or vein. The catheter is a functional instrument not only for vascular surgery, but also for invasive angiology. PMID- 12092128 TI - [Chorioamnionitis and early-onset neonatal sepsis do not significantly affect levels of interleukin-6 in very low birth weight neonates]. AB - OBJECTIVE: To determine the influence of maternal chorioamnionitis and neonatal sepsis on interleukin-6 (IL-6) levels in cord blood and in blood obtained from very low birth weight (VLBW) infants within the first two hours of life. DESIGN: Prospective clinical study. SETTING: Institute for the Care of Mother and Child, Prague. METHODS: We measured the serum levels of IL-6 in 30 consecutive VLBW infants born in our institute. IL-6 levels were evaluated in cord blood and in neonatal blood within 2 hours after delivery. Maternal chorioamnionitis and neonatal sepsis within the first 72 hours of life were monitored. RESULTS: Maternal chorioamnionitis was detected in 7 of 30 patients (23.3%). There was no significant increase in IL-6 level in cord blood of newborns with maternal chorioamnionitis (p = 0.42). Serum level of IL-6 in this group did not differ from the level in newborns of mothers without signs of intraamniotic infection (p = 0.39). Neonatal early-onset sepsis was diagnosed in 7 of 30 patients (23.3%). There was no influence of neonatal sepsis on IL-6 level in cord blood (p = 0.98) and IL-6 level in neonatal blood (p = 0.19). We did not find any correlation between the group "chorioamnionitis positive" and "sepsis positive" (p = 0.31). CONCLUSION: IL-6 in cord blood or in neonatal blood within 2 hours of life was not enough sensitive and specific marker of maternal chorioamnionitis as well as for early-onset neonatal sepsis in the group of very low birth weight infants. PMID- 12092130 TI - [Less common causes of thrombocytopenia (case reports)]. AB - We report three case reports of different, less frequent causes of thrombocytopenia--autoimmune thrombocytopenic purpura (AITP), thrombotic thrombocytopenic purpura (TTP) and heparin-induced thrombocytopenia (HIT). Pathophysiology of these conditions is also discussed. AITP is an autoimmune disorder characterized by destruction of thrombocytes by autoantibodies targeted to platelet antigens. The cause of thrombocytopenia in TTP is an endothelial disorder with subsequent formation of platelet aggregates. HIT type I is a frequent, mild form of thrombocytopenia caused by reversible aggregation of thrombocytes following to heparin administration. HIT type II is a late, more severe form of thrombocytopenia caused by immunologic mechanisms. PMID- 12092129 TI - [Heparin-induced thrombocytopenia in patients and prevention with low molecular weight heparin]. AB - Prospective study implicates problems of heparin-induced thrombocytopenia (HIT). This is one of the most important complications in heparin treatment and prophylaxis. The set was formed of patients with severe diagnosis and larger intra-abdominal operation that was associated with relative high risk of thrombotic event. That's why there was need of heparin prophylaxis. We used Enoxaparin in dose 0.2-0.4 ml subcutaneously. The controlled parameters were platelet counts, detection of antibodies using ELISA Assay method (GTI-PFA), indirect platelet immunofluorescent test for monitoring platelet disorders. We had no known case of clinical symptoms of HIT. The change in platelet counts was due to postoperative course, or binding on blood transfer or basic haematological disease. The test detecting antibodies was positive in eight percent without clinical correlation, without important thrombocytopenia. Low-molecular-weight heparin has certain advantages over unfractionated heparin, as well know from clinical and laboratory monitoring in other studies, including larger safety (in relationship with HIT) with very effective prophylaxis. PMID- 12092131 TI - [Conflicts regarding the Hospital for Epidemics in Olomouc 1925-1938] ]. AB - The argument between the municipality of Olomouc and the provincial government in Brno was caused by diverse opinions, if an epidemic hospital for the large region of the North-Eastern Moravia should have been built in Olomouc. The Czechoslovak health care laws needed rehabilitation and unification of norms and conditions for building, financing and managing hospitals, as well as modernization of institutions concerning infectious diseases. The Moravian authorities quarrelling on the financial issues of the projected hospital, and later on the technical aspects of it, had to ask the Ministry of Public Health and Physical Training in Prague to unravel the problem, but without an effect. The solution was not easily achievable until the era of antibiotics and buckshee and compulsory inoculation after the World War II came. Microbiologist Professor Jan Kabelik took part in both the battles--for the new hospital in Olomouc in the pre-war period as well as for the immunization of the whole populace after the war. PMID- 12092132 TI - [The gallstone and arterial hypertension]. AB - In this study, I analyzed arterial hyperpressure which I found in gallstone. OBJECTIVES: The incidence between arterial hyperpressure and gallstone. METHODS: From 1122 patients with gallstone--500 had arterial hyperpressure; these patients were examined: physical examination, abdominal ultrasonography and Ecg. RESULTS: In this study--44.56% had arterial hyperpressure. CONCLUSIONS: I appreciate that arterial hyperpressure coexist with gallstone. PMID- 12092133 TI - [Laparoscopic treatment of the ovarian cysts]. AB - Laparoscopy is widely used as a safety procedure in many clinical situations allowing diagnosis and/or surgical management in a minimally invasive fashion. Most laparoscopic cases allow patients to recover quickly. The conventional ultrasonography and endovaginal sonogram appear to be useful in the pre-operative selection for this surgery. Conventional pulsed Doppler (Duplex) and color Doppler sonography (CDS) have added a new dimension to pelvic ultrasonography. This study was undertaken to evaluate usefulness of operative laparoscopy in treatment of ovarian cysts. A retrospective study was carried on 35 operative laparoscopy cases performed between 1995 and 2000. Types of surgery performed were cystectomy, salpingo-oophorectomy, oophorectomy and wedge resection. The mean operation time was in the range of 65 to 120 minutes, mean while the mean postoperative stay was of 3,8 days. PMID- 12092134 TI - [Incidence of ocular injuries in women]. AB - The aim of study is to examine the occurrence of battered women seen in Clinica I Oftalmologie Iasi. It is a retrospective study for a 5 year period for documentation of injuries to women as a result of "domestic" violence. The patient range in age was from 15 to 90 years. Injuries ranged including laceration and haemophthalmos in most of the cases. In conclusion, the ophthalmologist must see "domestic" violence in these injuries. PMID- 12092135 TI - [The role of oxidative stress in normal and pathological adaptive reactions]. AB - After a brief presentation of the data concerning the mechanism of production of oxidative stress as an expression of cellular and molecular stressing aggression, the main actions and functional implications of the excess of free radicals at the level of the main tissues and organs are presented. Emphasis is made on the nuclear, cytosolic and membrane alterations produced by the lipid peroxidation phenomena and the cell protein degradation induced by the oxidative stress. Special attention is given to the beneficial and harmful properties of the free radical species of oxygen and nitrogen, beginning with their involvement in the production of the normal adaptive reactions and ending with their involvement in the pathogenesis of various pathological states. In the group of beneficial effects are included along with the antibacterial and antiviral properties of the free radicals and their participation in the autoregulation of the local arteriolar-capillary circulation network. As harmful properties are mentioned the cellular destructive phenomena produced by the free radicals, which are considered as the generators of a true radicals pathology (senescence, atherogenesis, cancerogenesis, neurodegenerative diseases, etc). PMID- 12092136 TI - [Primary obstructive megaureter- diagnosis and treatment]. AB - The authors have treated 13 cases of primary obstructive megaureter (POM) in children, presenting without vesicoureteric reflux. The diagnostic possibilities and the surgical approach options are discussed. Both Cohen and Leadbetter- Politano uretero-vesical reimplantation techniques proved to be successful in this series. Ureteric folding was done in 2 cases. Reported final results are good in 10 cases. PMID- 12092137 TI - The apoptosis of the rat and mouse testicular cells could be induced by synthetic analogous of the prostaglandin F2 alpha? AB - Numerous actions of the synthetic analogous of the prostaglandins were used in human and veterinary therapy. The action of these analogous (even, initially the prostaglandins were put in evidence in the seminal plasma) on the male reproductive system are little known. This is the reason for our study, which tried to emphasize the effects of analogous of the prostaglandin F2 alpha on the apoptosis of the rat and mouse testicular cells. Optic active Cloprostenol and Cloprostenol Isopropyl ester, in a dose of 100 micrograms/kg/day induces significant histopathologic modifications in the testis of rats and mice. These changes are evident after 14 days of treatment, but especially after 28 days of treatment. PMID- 12092139 TI - The prevalence of the owner of serologic markers for the hepatitis C virus, in a north-eastern territory of Romania. AB - The paper describes aspects in the natural evolution of hepatitis C virus (HCV) infection with severe outcomes induced by the typical or atypical clinical manifestations or chronic carrier state. In the interval 1995-1999 in the Iasi district morbidity from all types of viral hepatitis ranged from 153.4 cases per 100,000 inhabitants in 1995 to 66.6 in 1999. The features of A viral hepatitis epidemiological process was the multiannual cyclic pattern. From estimating the circulation of HCV in the population of Iasi district, several population categories have been screened for the presence of HCV antibodies (anti-HCV). Testing 87,894 blood samples assessed the prevalence of anti-HCV in blood donors. Different annual values resulted: 1.5% in 1997 and as a result of routine screening, only 0.3% in 1999. The cases of posttransfusion hepatitis (PTH) admitted to the Clinic of Infectious Diseases of Iasi, the diagnosis of which was supported by epidemiological history and confirmed serologically by the presence of anti-HCV were of 12 in 1995 (36.4% of all PTH), 20 in the interval 1996-1999, with an average for the 4 study years of 7.5%. Anti-HCV was found in 24.4% of series of 4,594 patients with viral hepatitis, in 32.1% of 582 individuals with various disorders or at risk, of which those with liver cirrhosis accounted for 37%, and in 9.6% of 94 patients with neurological disorders. Of the 1083 tested physicians and nurses working in health care settings at risk 6.6% were anti-HCV positive. PMID- 12092138 TI - [Interactions between liposomes and synaptosomes]. AB - In continuing experiments to determine the basis of presynaptic modulation, rat cortical synaptosomes were employed and transmembrane calcium fluxes were determined with a metalocromophoric dye, Arsenazo III. The intracellular free Ca2+ concentrations show complex fluctuations in time and space in response to a variety of stimuli, acting as a pluripotent signal for many neuronal functions. Depolarisation of synaptosomes by K+ 20 mM determined calcium influx in cortical synaptosomes, mainly mediated by the Q type Ca2+ channels, with a minor participation of the N type channels. The administration protocol for the polyamines, revealed that agmatine, spermine, spermidine, putresceine and cadaverine reduce, in variable proportions, the K(+)-induced Ca2+ influx, in synaptosomal preparations. The results plead for the ability of agmatine and spermine to modulate Ca2+ fluxes in synaptosomes, both from the interior and exterior side. Moreover, rigidization of the synaptosomal membrane (by cholesterol-enriched liposomes administration) do not significantly influences K(+)-induced Ca2+ influx in synaptosomal preparations. PMID- 12092140 TI - Study of outer membrane proteins (OMPS) and their role in co-modulation of resistance in non typhoidic Salmonella strains isolated in Iasi. AB - OBJECTIVES: To characterize the OMP profile and to study the possible interference with other resistance determinants. MATERIAL AND METHODS: 16 non typhoidic Salmonella strains, isolated in 1999 from stools of pediatric patients, were selected according to their resistance phenotype: resistance to Ampicillin (AMP), Amoxycillin/Clavulanic Acid (AMC), and third generation cephalosporins: Ceftriaxone (CRO) and Ceftazidime (CAZ). Identification and sensitivity testing were done by DADE MicroScan System. beta-Lactamases were characterized by isoelectric focusing (IEF) with preformed minigelss. OMPs were studied on membrane preparations on SDS-urea gels. RESULTS: Isoelectric points: Majority of the isolates had the association 5.4 +/- 7.6 (3/16) or 5.4 + 8.2 (6/16). The second beta-lactamase has the capacity to hydrolyse CAZ. We found three different patterns of OMPs: I: 30, 29 and 25 kDa; II: 30, 26 and 24 kDa; III: 30 and 25 kDa. We couldn't find any correlation between the OMPs profile and resistance phenotype, showing that the beta-lactamase production is the only resistant determinant. CONCLUSIONS: The study shows the high frequency of extended-spectrum beta-lactamases (ES beta LA) amongst the non-typhoidic Salmonella; association of TEM type and ES beta LA enzymes diminishes considerably the therapeutic resources: inhibitor associated combinations are non effective. In non-typhoidic Salmonella, porins seems to have no influence in co-modulation of resistance. PMID- 12092141 TI - [Detection of extended-spectrum beta-lactamases production in enterobacteriaceae strains]. AB - Extended-spectrum beta-lactamases (ESBL) cause resistance to ceftazidime, ceftriaxone, cefotaxime and other cephalosporins. The ESBL enzymes are most often derivatives of beta-lactamases from the TEM or SHV group. The aim of our work was to study ESBL production by 35 ceftazidime-resistant strains. We selected these strains among 335 Enterobacteriaceae, isolated in Eastern Romania from healthy carriers and patients, between january-june 1999. To identify the studied strains we used API 20E and API 32E. The ESBL production was established by using both double disk synergy test (DDT) and EXPERT computer program of miniAPI (Bio Merieux). We revealed ESBL production in 26 strains when EXPERT program was used and only 23 when DDT was used. Our results suggest that miniAPI is a better method to study ESBL production than DDT. PMID- 12092142 TI - [The para-clinic investigation of temporo-mandibular joint changes in patients with acromegaly]. AB - The Acromegaly is an endocrinological disease determined by the hypersecretion of STH in a certain period of the body evolution and it causes the hypertrophy of bones in general and of mandible and cranio-facial bones, determining a disorder due to this development of bones, associated with troubles in the activity of muscles and of the phospho-calcium metabolism. This study was made on a group of 33 acromegaly patients. Their temporo-mandibular joint was investigated by ortopantomography, tomography, computer tomography and scintigraphy. All of these exams shows the changes in temporo-mandibular joint due to the cells hyperactivity determined by the hypersecretion of STH. PMID- 12092143 TI - [Periodontal tissue changes after the correction of reverse over-jet]. AB - Frequently, the incisor crossbite is the result of the abnormal retroclination of the anterior teeth of the maxilla. The occlusal trauma due to this condition and crowding can affect the periodontal health of the anterior teeth. We took in our study a sample of children with incisor crossbite who have undergone orthodontic treatment with proclination of the maxillary incisors in order to correct the reverse overjet. All subjects were monitored for oral hygiene during the correction of the incisor crossbite. When found that the orthodontic movements for the correction of the incisor crooss-bite produced an increased and transcent periodontal inflammation in the maxillary incisor's area, but with a good oral hygiene, these kind of orthodontic movements seemed to be harmless for the periodontal status of this patients. PMID- 12092145 TI - [Analysis of attitudes about abortion]. AB - Today two clear trends can be discerned in the development of attitudes about abortion over the last century or so. The first corresponds to that category of "bio-power" described by Foucault as a bio-politics of the population. The second trend, closely related, concerns the transformation of abortion from a largely criminal to a largely therapeutic procedure, that is, one which is said to be performed in the interests of preserving or enhancing the health and well-being of the woman or her existing children. This paper analysis the attitudes about abortion exploring the responses of the Medical Bioengineering Faculty's students to a series of questions about the conditions under which legal abortions should be available and tried to identify some of the reasons why abortion attitudes vary from person to person. PMID- 12092144 TI - [Some aspects concerning the electrochemical corrosion of the Gaudent-S dental alloy]. AB - The potentiodynamic polarization methods have been employed to evaluate the corrosion resistance of the Gaudent-S dental alloy in a slightly acid artificial saliva (Fusayama's saliva; pH = 6.18), a slightly alkaline artificial saliva (Rondelli's saliva; pH = 7.75) and in an acid saline solution (NaCl/lactic acid; pH = 2.64). The results were compared with those obtained for copper and aluminium--the principal components of the studied alloy. In Fusayama's artificial saliva both the copper and Gaudent alloy exhibit close indentic cyclic voltammograms, typical for pitting corrosion; the pitting potentials being 1550 mV (SCE) for Gaudent-S and 1350 mV (SCE) for copper, while the repassivation potential is 700 mV (SCE), the same for the two analysed metals. In the Rondelli's artificial saliva and in acidic saline solution the passivation domain is considerable reduced; the corrosion potential and the repassivation potential coincides. In the saline acid solution the current densities are two times higher than those obtained in the other two solutions. In the aluminium case it can be noticed that the passivation domain is very reduced, no cathodic peak is observed and much more current densities are registered in the positive potential domain. PMID- 12092146 TI - [CA 15-3 and CEA markers - prognostic factors in breast cancer]. AB - For the increasing of sensibility and specificity in the monitoring of the diagnosis and treatment of breast cancer it is used the simultaneous determination of two tumoral markers: CA 15-3 and CEA. This study try to demonstrate the grade of involvement in diagnostic and screening. We have determinate CA 15-3 and CEA in 73 serum samples. The correlation between the stage of malignancies and CA 15-3 levels was relieved in the second section of this study. The third section follow up the dynamic evolution of CA 15-3 marker in pre- and postoperator (14 days) stages obtaining a decreasing of CA 15-3 level. The obtained results for CEA does not establish any clinical rules in order to compare with the CA 15-3 level and disease stages. The easy-used and non invasive determination, the early increasing of it's level and it's preservation recommend glycoprotein CA 15-3 like an current clinical instrument. PMID- 12092147 TI - [Ultrasonography's role in prenatal diagnosis: The nuchal fold]. AB - Two cases of nuchal fold abnormalities diagnosed by transvaginal ultrasonography are presented: nuchal edema at 10 weeks (fold = 5.5 mm) of normal size at 13 weeks (fold = 2.6 mm) and cystic hygroma colli associated with omphalocel in a women with a 24-week arrested pregnancy terminated by spontaneous abortion. Genetic tests done in the second case showed a karyotype 45XO. The etiopathogenesis of nuchal skin fold abnormalities--edema and cystic hygroma colli is discussed. The importance of ultrasound screening during pregnancy is emphasized, the detection rate of genetic abnormalities depending on both the quality of the ultrasound equipment and examiners skills. Ultrasound examination at 10 and 20 weeks--ultrasound marker (nuchal fold, LF ratio and DBP/FL ratio)- together with the triple test (AFP, HCG and E3) in the mathematical model used for detecting the risk of genetic defects may represent an alternative to amniocentesis and trophoblast biopsy when this is impracticable or rejected by the patient. PMID- 12092148 TI - Original method for nicorandil synthesis. AB - Nicorandil (N-(2-hydroxy-ethyl)-nicotine amide nitrate), drug recently introduced in therapy, is used as a vasodilator through potassium channel activation. Nicorandil is obtained by treating nicotinic acid chloride hydrochloride with nitro-oxy-ethyl-amine nitrate in an organic solvent. The authors suggest a method that uses ethyl nicotinate as raw material. Thus, the drug can be obtained from indigenous raw materials, at a lower price than the imported ones, with a high purity and a good yield. PMID- 12092149 TI - [Biofeedback therapy for constipation and fecal incontinence]. AB - Constipation and fecal incontinence are problems that are frequently encountered in daily practice. Over the last decade biofeedback therapy has emerged as a useful adjunct for the treatment of these problems. Biofeedback therapy is a learning process that is based on "operant conditioning" techniques. Biofeedback training aims to restore a degree of continence that enables patients to resume their activities to a satisfactory level with good success rates and prolonged benefits. The technique requires height levels of motivation not only in the patients but also in the instructor who is providing verbal feedback about the adequacy of a patients performance. PMID- 12092150 TI - [Endocrinology at the beginning of the millennium. The seventh congress of the Romanian endocrinologic society. Iasi, 27-30 September, 2000]. PMID- 12092151 TI - [In memoriam of Ignaz Phillip Semmelveiss, initiator of asepsis and antisepsis, at 135 years of his death. Sepsis and its prevention]. PMID- 12092153 TI - [History of orthopedics and traumatology]. PMID- 12092152 TI - [NK cells in the physiology of the fetomaternal relationships]. AB - The NK cells are large granular lymphocytes that are found as approximately 2-15% of the global peripheral lymphocytes. As endometrial granulocytes, they represent 70% of the decidual leukocytes during the first trimester of pregnancy, with a progressive decreasing until term. The complex actions of these numerous and active cells, consist in: 1) placental regulation and control, based on the receptors for the HLA--G and HL--C antigens, that are trophoblast specific and act as universal inhibitors of the NK cells; 2) secretion of a large range of cytokines as IL-1, TNF alpha, IL-8, TGF, IFN gamma, GM-CSF, CSF1 which regulate the trophoblastic proliferation, differentiation and invasion; 3) removal of abnormal trophoblastic cells, which may appear in such a proliferating cell population; 4) infectious protection of the placental unit, due to their unspecific cytotoxic effect. In conclusion, the NK lymphocytes have important implications in the formation of the decidua, as the maternal side of the fetomaternal interface and they are deeply involved in the evolution of normal pregnancy concerning the nutritional, hormonal and immunologic aspects. PMID- 12092154 TI - [Molecular mechanisms of apoptosis. Fas and TNF systems. ICE protease system. Bcl 2 family]. AB - For every cell is a time to live and a time to die. The cell death that occurs during embryogenesis, normal tissue turnover, pathological conditions like cancer cells, etc. is "programmed cell death" or apoptosis. In this study we report on apoptosis mediated by cytokines. Fas ligand and tumor necrosis factor bind to their receptors and induce apoptosis. PMID- 12092155 TI - [Recent data about the LDL-atherogenesis relationship]. AB - The steps in pathophysiology of ATS make easier the understanding of waves of processes that lead to the disfunction of the endothelial cells, hypertrophy of the arterial wall and in the end to the complication which drawn from this process. Atherogenic process which is characterised by proaterofenethic and atherogenetic mechanism imbalance suppose three phases: early, transition and terminal phase. The initial phase involve the focal influx and LDL-cholesterol accumulation in the arterial intima, the ROS (reactive oxygen species) and azot generation by smooth muscular cells, macrophages and endothelial cells and oxidative modification of LDL. Scientific data revealed that LDL oxidation is mediated by peroxinitrite (ONOO) anion which could act directly over the thiol groups or indirectly through OH and NO radicals from peroxinitrous acid breakdown and who initiate the polyunsaturated fatty acids peroxidation and the oxidation of apolipoprotein B-100. LDL completely oxidated are recognised by scavenger receptors which are not regulated by up and down regulation. The last event of the early atherogenic process is the formation of foam cells from macrophages through LDL modified captation by scavenger receptors. PMID- 12092156 TI - [Diagnostic strategies in Hepatitis C virus infection]. AB - Hepatitis C virus (HCV) is the most common cause of chronic viral hepatitis. The World Health Organization estimates that 170 million people world-wide are infected with HCV; 70% of them will develop chronic hepatitis and 20-30% cirrhosis in 10-30 years. Of those with cirrhosis, an estimated 25-30% will develop liver cancer. Since the identification and molecular characterization of HCV in 1989, a variety of diagnostic tests based on the detection of hepatitis virus antibodies or HCV RNA in the serum have been developed. The enzyme-linked immunosorbent assays (ELISA 3) and the recombinant immunoblot assays (RIBA 2nd and 3rd generation) exhibit improved sensitivity and specificity for HCV antibodies. Qualitative and quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) has allowed clinicians to track the natural history of HCV and to monitor the progress of therapy. This article reviews the state-of-the-art tests and assays developed for the diagnosis and management of HCV infection. PMID- 12092157 TI - [Gastroesophageal reflux disease in pregnancy]. AB - Gastroesophageal reflux disease (GERD) symptoms are common in pregnancy, occurring in approximately 45% to 80% of pregnant women. Although the symptoms associated with reflux in pregnancy are similar to those described in the nonpregnant state, some of the etiologies are distinct due to hormonal fluctuations and other physiologic changes often associated with pregnancy. Diagnostic tools and therapeutic regimens that might be used without hesitation in the nonpregnant patient must be given with cautious consideration in the gravid patient due to potential fetal risks. Pregnant patients with symptomatic GERD should be managed aggressively with lifestyle modification and dietary changes. Antacids and antacids/alginic acids combination or sucralfate should be considered first-line medical therapy; treatment with cimetidine or ranitidine should be considered; these H2 receptor antagonists are preferred during pregnancy. Proton-pump inhibitors should be used with caution because little human experience is available. PMID- 12092159 TI - [Locally advanced breast cancer in the elderly]. AB - The data concerning the influence of old age and locally advanced stages on breast cancer treatment and outcome are lacking or sparse. The authors reviewed treatment modalities in women over 65 years old with locally advanced breast cancer. The purpose of the study is to determine if differences in treatment modalities, clinical stage and pathological type influence the outcome. For patients with locally advanced disease but without over evidence of metastatic disease, radical surgical procedures should be attempted. Most study results confirm that loco-regional therapy and adjuvant chemohormonotherapy is feasible and safe even for older patients. PMID- 12092158 TI - [Cigarette smoking and lung cancer-elucidating an enigma]. AB - The complexity of tobacco smoke leads to some confusion about mechanisms by with is caused lung cancer. Tobacco smoke contains many carcinogens, but 20 of them are deeply involved in causing lung cancer induction. Of these, polycyclic aromatic hydrocarbons and tobacco specific nitrosamine are likely to play a major role. This review include a discussion of the mechanisms of interaction between DNA and tobacco smoke carcinogens and the relationships with mutations in oncogenes and tumor suppressor genes. By focusing in this review on several important carcinogens in tobacco smoke, the complexities in understanding tobacco induced cancer can be reduced, and new approaches for lung cancer prevention can be envisioned. PMID- 12092160 TI - [Portal hypertension in liver cirrhosis]. AB - Portal hypertension is a dramatic complication in liver cirrhosis by efraction of the varices localized more frequently in the esophagus. It has been an actual subject due to physiopathological research (endotelin system, nitric system etc), diagnosis (echoendoscopy, color Doppler), and therapeutically progress. Propranolol, available at a low cost, is efficient and unanimously accepted in the prophylactic treatment of medium and large varices which have never bled as well as in the hemorrhage due to variceal efraction which stopped therapeutically or spontaneously. This drug diminishes the risk of rebleeding. Besides the pharmacological treatment, terlipresine, octreotid, sclerotherapy and bandlegation are used in active hemorrhage due to variceal efraction. If this fails other methods for haemostasis are used: portosystemic transjugular shunt, mechanical tamponade, selective surgery and ideally hepatic transplant. Except for the hepatic transplant, none of the mentioned methods can improve the rate of survival in-patients, which depends on the state of the hepatic failure. PMID- 12092162 TI - [The beginning of the year, the century and the millennium]. PMID- 12092161 TI - [Actual modalities of separation by magnetic methods of active biological compounds, cells and pathogenic agents]. AB - The paper presents an overview on magnetic techniques of bio-processing and some of their possible applications. These techniques are in fact direct, or more frequently, indirect magnetic means for separation of particulate substances. A brief presentation of direct and of the most important indirect methods (magnetic carrier and magnetic fluids technologies) is made. There are shown some of the possible applications of magnetic bio-processing in bio-technology and medicine: active biological compounds fixation, isolation and modification, cells and cells organelles separation, removal of xenobiotics, immuno-magnetic testing, pathogen microbes identification. PMID- 12092163 TI - [Myocardial bridging and sudden death]. AB - Myocardial bridging (MB) has been described more than 200 years ago. However it's implications on the genesis of myocardial ischaemia were not studied until recently. Little is known about the real incidence, survival in people with this entity and pathophysiology. Data found in the literature are prone to bias since all studies published are retrospective and the populations studied are limited and very selectioned. Most authors agree that most MB is rather frequent and seldom generates ischaemia. A significant clinical effect of myocardial bridging implies a thick MB with at least 75% coronary systolic obstruction associated with ventricular hypertrophy and/or rapid tachycardia. In the peculiar case of sudden death, we think that to consider MB as a cause of this sudden death we need more than a simple necroptic finding of a MB. A history of clinical and/or electrocardiographic confirmed ischaemia or the presence of histological abnormalities suggesting myocardial ischaemia can be helpful. PMID- 12092164 TI - [Age related electrophysiological modifications in rat myocardium]. AB - During aging the electrical and mechanical phenomena of the heart are impaired. We studied the effect of aging using standard microelectrode technique on action potentials obtained in papillary muscle from 1, 3, 6 and 18 month old rats. The action potential of myocardial cells is significantly prolonged and changes resemble those observed during cardiac overload. The possible cause of the slower action potentials is the alteration of Ca2+ transients due to a reduced rate of Ca2+ sequestration by the sarcoplasmic reticulum associated with a possible alteration of K+ currents. These developmental changes are considered as an expression of adaptation of the heart with aging. PMID- 12092165 TI - [The role of exercise testing in evaluation of functional capacity at patients with ischemic ventricular dysfunction]. AB - The assessment of functional capacity represents an important parameter in the prognostic stratification of patients with heart failure. The objective of the paper is the testing of exercise capacity at patients with ischemic ventricular dysfunction. METHODS: There have been selected 80 patients with coronary heart disease (old myocardial infarction, angina pectoris, ischemic cardiomiopathy) admitted in the Department of Internal Medicine, Rehabilitation Hospital, Iasi, during 1.01.1998-31.12.1999. They were divided into two groups: group A (46 patients) with proper ventricular function and ejection fraction (EF) above 50% and group B (34 patients) with EF under 50%. They underwent exercise testing at the ergometric bicycle. RESULTS: Patients of group B had higher values of resting heart rate (74 +/- 9 vs 70 +/- 11, p = 0.04). There were registered similar levels at the maximal exercise at both groups (time-tension index, watt, MET). The evolution of the group with ventricular dysfunction is distinguished by a lower length of time of exercise (8.6 +/- 4.4 vs 9.8 +/- 5.2 min, p = 0.05) and by a significantly greater value of the ST depression (1.6 +/- 1.4 vs 0.9 +/- 1.1 mm, p = 0.01). DISCUSSION AND CONCLUSIONS: The higher resting heart rate at patients with heart failure suggests the activation of neuroumoral mechanisms, especially the adrenergic activation. These patients reach maximal levels similar to those with good systolic function but in a shorter length of time, due to a physical deconditioning or to the same neuroumoral mechanisms. Ischemia is higher at these patients. PMID- 12092167 TI - Liver transplantation: ethical issues. PMID- 12092166 TI - [Thyroxine therapy in benign thyroid nodules]. AB - Levothyroxine (LT4) treatment in benign thyroid nodules is a controversial management. The favorable response varies between 10-60%, being, in some studies, comparable between treated and untreated groups. The aim of this study was to evaluate the response of uninodular goiter at the LT4 treatment, in comparison with untreated patients. The study group (S) included 53 patients diagnosed with nodular goiter, treated with LT4 for 1 year. 26 patients with nodular goiter, age and sex-matched, untreated, constituted the control group (C). All patients were from a minor iodine deficient area. The including criteria were euthyroidism, single nodule, solid (ultrasonography), cold (Scintigraphy), and benign (FNAB). After 1 year mean nodular volume had a significant decrement in both groups, higher in group S (35%: from 7.8 to 5.2 mL, p = 0.0098) than in group C (25%: from 8.4 to 5.9 mL, p = 0.026). Linear regression showed a slight correlation between the nodular decrement and the initial volume (r = 0.23): the responders percentage was higher in nodules with a volume < 5 ml than in those with volume > 5 ml (51.5% vs o 19.6%, p < 0.0001). The evolution of treated nodules seamed to be better than of the untreated ones, but the differences were only slightly significant. We believe that the evolution of thyroid nodules under LT4 treatment can be influenced by the iodine supply, since in most of the studies from minor iodine deficiency regions (Europe, South America) the response is better than in regions with sufficient iodine supply (North America). Knowing the potential side effects of LT4 therapy, this kind of management of benign thyroid nodules should be reserved to selected cases. PMID- 12092168 TI - [Lymphocyte traffic in the events that follow lymphocytes activation in the germinal centre]. AB - After their maturation, lymphocytes migrate from the primary lymphoid organs to the secondary lymphoid organs and tissues. Once in the secondary tissues the lymphocytes do not simply remain there; many move from one lymphoid organ to another via the blood and lymph. This process allows a large number of antigen specific lymphocytes to come in contact with their appropriate antigen in the microenvironment of the peripheral lymphoid organs and this is important since lymphocytes are monospecific and only a limited number of cells are able to recognize and interact with a particular antigen. When B cells are activated by antigen, with the help from the T cells, they mature either into AFCs (Antibody Forming Cells), or they develop into memory cells. The germinal centers in various lymphoid tissues represent the sites of the immune response development and memory B cells generation. At these sites, the B cells undergo active hypermutation of the variable genes, a process that can lead to death by apoptosis for some cells. PMID- 12092169 TI - [Neurogenetics in Parkinson's disease]. AB - The pathogenesis of Parkinson's disease, a progressive degenerative disorder of the central nervous system, remains obscure. 20% of cases are familial. The disorder may be due to a combination of genetic and environmental factors. Some monogenic forms of Parkinson's disease have been described. Many genetic studies explore whether a candidate gene may confer an increased risk for this disease. The search for a genetic susceptibility may lead to establish a preventive therapy. PMID- 12092170 TI - [Imidazoline receptors-normal and pathological factors]. AB - Imidazolinic receptors I1 and I2A and I2B different from alpha 2 receptors and by any other type of receptors were identified in the last years in many tissues from human body and in cells of a great number of animals. There are agonists and antagonists on I1 receptors (e.g., agonists like moxonidine, rilmenidine and antagonists like efaroxan, BAF 6143 etc.) and also on I2 receptors (e.g., agonist idazoxan, etc.). There are many actions performed by stimulation of imidazolinic receptors like: decreasing of blood pressure (I1 stimulation from midbrain), decreasing of noradrenaline plasmatic level, antiarrhythmic actions in experimental arrhythmias etc. Imidazolinic receptors are involved in natriuresis, pancreatic insulin production, some superior nervous activities, regulation of food intake etc. We consider that imidazolinic receptors and their endogenous agonists (e.g. agmatine) represent an important system involved in modulation of cell function. PMID- 12092172 TI - Gastroparesis and its management. AB - Gastroparesis may be related to a variety of underlying disorders, but diabetes mellitus is by far the most common cause. Symptoms of gastroparesis include early satiety, postprandial bloating, nausea and vomiting. Gastric scintigraphy with 99 technetium-labeled low-fat meal is the gold standard method of diagnosing delayed gastric emptying. Dietary measures and prokinetic drugs bring symptomatic relief in most patients. Some patients with severe nausea and vomiting will require antiemetic medications. Few patients will fail medical therapy and will continue to have debilitating symptoms of gastroparesis; such patients may benefit from a venting gastrostomy or a jejunostomy placed surgically, endoscopically, or fluoroscopically. Gastric electrical stimulation is an exiting new approach in the management of gastroparesis. As the treatment of gastroparesis is far from ideal, nonconventional approaches and nonstandard medications are presented. PMID- 12092171 TI - [Opioid tolerance and dependence--pharmacological aspects]. AB - Prolonged opioids administration leads inevitably to tolerance and dependence, a phenomenon we meet more often in healthy people than in ill patients. Tolerance means a hypersensibility of neuronal membranes as well as changes in the number and affinity of opioid receptors, which implies intake of larger doses to obtain the initial effect. Physical dependence, quite different of the psychological one, is the appearance of abstinence syndrome on sudden interruption of opioid administration or on administration of an antagonist. There is usually cross tolerance in opioids, but it can also be incomplete, when the initial opioid can be replaced with another one that produces a milder abstinence syndrome. Classically, metadone is used in long time therapy, after detoxification with an antagonist is performed (naloxon, naltrexon). Modern pharmacological alternatives are levo-alpha-acetyl-methadol (LAAM) and agonists-antagonists (butorphanol, buprenorphine, pentazocine, nalbuphine). An antagonist can also be used if associated with an alpha--stimulant (clonidine), in order to remove noradrenergic manifestations of abstinence syndrome. Now other therapeutical principles are being studied: enkephalinaze inhibitors to reduce the abstinence syndrome, NMDA receptor antagonists, NO sintetasis inhibitors, that facilitates opioid analgesia and hinders tolerance development; colecystokinin-receptors agonists or antagonists to reduce tolerance on morphine. A recent study showed that the concomitant administration of an opioid agonist (sufentanil) and a calcium channels blocker (nimodipine) not only prevents from tolerance development but also triggers hypersensibility to analgesic effects of the opioid. PMID- 12092173 TI - [White-Bland -Garland syndrome: the maximum natural development of the coronary anastomosis ]. AB - The anormalous origin of the left coronary artery from pulmonary artery (White Bland Garland syndrome) is a very rare coronary malformation, having despite the great mortality in the early childhood, an adult form, characterized by minor symptoms and long course. This paradoxical situation is totally dependant to the complete development of the coronary anastomosis, allowing the right coronary flow to perfuse the left myocardium. The theory of the "terminal arterial perfusion" of the heart, still persistent in some monographs is meeting in this natural situation its strongest opponent argumentation. The coronary anastomosis, clearly visible by coronarography are large enough (in late stages) to inverse the flow in left coronary artery, filling the pulmonary artery. Is the coronary pulmonary shunt, responsible for the majority of symptoms, in the 5th or 6th decade. The adult form of the White-Bland Garland syndrome is indicative for the maximum result we may hope to reach using the new revascularisation method for ischemic heart disease: new vessels formation and enhancing the coronary anastomosis by growth factors administration therapy. PMID- 12092174 TI - [Clinical and biochemical aspects of diabetic nephropathy]. AB - Diabetic nephropathy (DN) is the most important cause of increased morbidity and mortality in patients with diabetes mellitus. Moreover DN is associated with a high risk for cardiovascular complications and progression of renal failure. It is known that there is a cumulative risk of development of DN represented by genetic factors, glycaemic factors, hypertension, alterations in the lipid metabolism, smoking. Functionally, it is represented by increased glomerular filtration rate (silent phase) and renal hypertrophy, increased the urinary albumin excretion rate, increased blood pressure and decline in glomerular filtration rate (end phase). Biochemical DN induced the alterations in lipid metabolism (increased the serum concentrations of triglicerides, VLDL, LDL), alterations in glycaemic metabolism (blood glucose, glycated hemoglobin, glycated albumin). The correlations between clinical and biochemical aspects are developed in this reference. PMID- 12092175 TI - [The influence of body weight upon the function of ovarian axis]. AB - The relationship between the body weight and the function of hypothalamopituitary ovarian axis was longtime studied. Frisch and Ravell (1971) have proposed the hypothesis that the onset of menarche is strong related to the achievement of a critical body weight. These authors observed that, despite the decrease in the last 120 years of the menarche age from 16.5 to 12.5 year-old, the body weight at which the menarche appears remains unchanged, 47.5 +/- 0.5 Kg. Many studies show the importance of both, body weight and fat mass percentage, in the appearance of menarche at puberty, or in the restoration of menses after the weight loss amenorrhea. Primary or secondary underweight amenorrhea can be associated to an eating disorder (anorexia nervosa, bulimia nervosa, or the alternation of these to clinical conditions), to severe exercise (athletes, gymnasts, dancers) or to malnutrition. The connected signal between metabolic status and reproductive function may be represented by the substances like: insulin, amino acids, IGFPB I, leptin. The low levels of leptin were found in underweight female with oligo or amenorrhea. By the other hand, obesity is not a primary factor causing chronic anovulation. However, obesity may aggravate an already existing subtle defect in some women and result in amenorrhea. PMID- 12092176 TI - [Renal cancer :therapeutical dilemma]. AB - Renal cell carcinoma constitutes 3% of all adult malignancies. Surgical resection remains the cornerstone of management for localised renal cell carcinoma. No effective postsurgical adjuvant therapy has been established for patients with locally advanced disease who are a high risk for recurrence. The effective treatment of metastatic kidney cancer remains a challenge. Despite extensive investigations with different treatment modalities, metastatic renal cell carcinoma remains high resistant to systemic therapy. Combination chemotherapy alone or in combination with cytokine, is a very little use. Small numbers of patients' exhibits complete or partial responses to interferon and/or interleukin 2, but most patients do not respond and few survive over the long term. New immunologic approaches to the treatment of both advanced and high-risk postsurgical disease are focusing on novel vaccine therapies to target both renal epithelial and vascular antigens. PMID- 12092177 TI - [Fever of unknown origin : infectious etiology and diagnosis principles]. AB - The paper has 3 parts: definitions, causes and diagnosis principles in prolonged fevers of infections etiology. The non infections causes of FUO are only mentioned. The infections causes are presented as: Bacteria, Viruses, Mycetes and Protozoa. The part regarding the etiological diagnosis firstly includes data about the microbiological diagnosis methods and samples in four steps: a) routine samples done for every febrile patient in a general hospital; b) all the accessible samples and c) samples obtained by invasive procedures (needle biopsies and open biopsies). When all of these had negative results in microbiological tests, a last possibility is adopted: exploratory laparatomy, eventually by sampling some specimens. Finally, conditions of obtaining interpretable serological tests are listed. PMID- 12092178 TI - [Bartonella infection]. AB - Bartonella species have been recently recognized as an important human pathogen associated with a wide spectrum of diseases. Four members of the genus are known to cause human infection: Bartonella baciliformis, B. henselae, B. quintana and B. elizabethae. B. baciliformis, the first identified Bartonella species, is the agent of two disease entities, Oroya fever and verruga peruana., B. henselae and B. quintana are two species involved in producing bacteremic syndromes (relapsing fever, trench fever, endocarditis), chronic lymphadenopathy in immunocompetent patients (cat-scrath disease) and chronic vascular lesions in immunocompromised hosts (bacillary angiomatosis and bacillary peliosis hepatis-recognized as new opportunistic infections in HIV-infected patients). PMID- 12092179 TI - [Immunoisolation strategies for the transplantation of pancreatic islets]. AB - Several different types of systems employing selectively permeable membranes and matrix supports for cells have been successfully tested in animals. Results in diabetic animals indicate that these systems can function for periods of several months to several years without the use of any of immunosuppression. This approach has the potential not only to allow the transplantation of islets across wide species barriers but that it can be achieved using injectable microreactors fabricated from biodegradable polymers. A new prototype of minimal volume alginate/polyaminoacidic microcapsules which retain immunoisolatory, biocompatibility and functional properties that seem to match those of conventional-size microcapsules was developed. Since coherent microcapsules tightly envelop each islet, any redundant space between membrane and islet is virtually eliminated. Consequently, these microcapsules occupy an extremely this space, thus addressing a major problem associated with conventional size microspheres. PMID- 12092180 TI - TNF alpha and IL-6 involvement in surgical trauma. II. In vitro cytokine production. AB - The study aimed to correlate TNF alpha and IL-6 dynamics during surgical trauma as well as the changes which appear in the circulating monocytes' response after LPS stimulation in vitro. Three patients with major abdominal operations were chosen and serial blood samples were taken before, during and after the operation. Normal unmatched individuals were used as control group during a two hours bed resting period. Both TNF alpha and IL-6 in vitro production showed similar patterns with an initial drop and almost full recovery 24 hours after the operation. We propose a model for TNF alpha and IL-6 role in trauma induced inflammation. PMID- 12092181 TI - Antioxidant enzymes levels in children with juvenile rheumatoid arthritis. AB - Pathogenic mechanism of chronic inflammation is associated with increased production of superoxide anion and hydrogen peroxide. In the neutralization process of that anions, superoxid dismutase (SOD), catalase (CAT), and glutation peroxidase (GPx) are key enzymes. Aim of study consists of establishing of some clinic-biological correlations in JRA chronic inflammation in childhood between clinical status and determination of lipoperoxidation products and antioxidative enzymes in the blood. MATERIAL AND METHODS: Blood samples were obtained from 20 patients admitted in 2nd Clinic of Pediatrics, 4-6 months after onset of disease, diagnosed with JRA, oligoarticular form (6 cases), poliarticular form (9 cases) and systemic form (5 cases), as compared to 10 control subjects. SOD, CAT, GPx were measured comparing with malonildialdehyde (MDA), seric glutation (GSH) and usual inflammatory tests (ESR, fibrinogen, CRP). Determinations were repeated after 6 weeks of treatment. RESULTS: In all our cases, level of antioxidant enzymes (CAT, GPx) was decreased at time of diagnosis, concomitant with increased MDA, SOD and inflammatory tests. In most of cases, after 6 weeks of correct anti inflammatory treatment, levels of enzymatic antioxidant markers were still decreased, as compared to usual inflammatory tests that came back to normal. Persistent decreased antioxidant enzymatic activity was found in cases that need immunomodulatory activity (Methotrexat). CONCLUSIONS: Determination of antioxidant enzymes level can be considered an evolution marker in JRA. More studies are necessary to find if antioxidant potential of blood can be used as following marker for immunosuppressive therapy. PMID- 12092182 TI - [Enzymatic markers in the alcoholic cardiomyopathy]. AB - Alcoholic cardiomyopathy can ensue from heavy consumption of alcohol over a long period of time. The clinical features include dilatation of the left ventricle, poor myocardial contractility with reduced left ventricular ejection volume, raised tissue enzymes. In numerous experimental data has been observed increased generation of oxygen and ethanol free radicals, indicate that free radicals are implied in myocardial and hepatic damage. Ethanol administration also elicits hepatic disturbances in the availability of antioxidant defense. The resulting antioxidative stress leads to enhanced lipid peroxidation and can also affect other important cellular component. PMID- 12092183 TI - [The histopathological study of the bronchopulmonary carcinoma]. AB - PURPOSE: The aim of the study is to reveal the predominant histological types of the bronchopulmonary carcinomas diagnosed at the Pneumology Hospital in the period of 1st January 1996-December 31 1998. MATERIAL AND METHODS: The study enclosed 196 cases of CBP, representing 174 men and 22 women with ages between 31 and 78 of years. The studied specimens were pieces of bronchoscopical and surgical biopsies, material obtained from the pneumological clinic. RESULTS AND DISCUSSIONS: The histological analysis revealed the increasing number of the bronchopulmonary carcinoma, mainly to men with mean age of sixty and the predominance of the epidermoid carcinoma (137) and a reduced number of cases of adenocarcinomas (31) and undifferentiated carcinomas (28). The knowing of the histological type of the CBP is very important in the calculation of the prognosis. CONCLUSIONS: Our study demonstrate the predominance of CBP at men (174), with ages between 55 and 65 years (127 cases) and similar incidences for all histological types of CBP comparative with other territories. PMID- 12092184 TI - Actinic keratosis--a histoenzymological study. AB - Actinic keratosis or senile keratosis is the most frequent premalign epithelial lesion that occurs in the elderly with a history of prolonged and intense exposure to the UV radiation and with an inborn susceptibility. Although the genetics and risk factors are clear nowadays, the histogenesis of this lesion is still under study. The histoenzymologic tests (such as those that demonstrate the ATP-ase and SDH-ase activity) are very useful for early detection of the structural and biochemical changes within the actinic keratosis lesion. PMID- 12092186 TI - [Antenatal corticosteroid therapy and the effects on complications of prematurity]. AB - The respiratory distress syndrome (RDS) in the preterm neonates is the most frequent cause of morbidity and mortality in neonatal units. Corticosteroids were shown to have multiple beneficial effects in maturing fetal surfactant and in decreasing the incidence and severity of RDS, intraventricular hemorrhage and necrotizing enterocolitis. MATERIAL AND METHODS: The authors studied 65 premies treated antenatally with corticosteroids versus 68 preterm neonates that didn't benefit from this therapy. All cases presented life threatening distress by various causes. There were evaluated the incidence of hyaline membrane disease, incidence related with gender, gestational age, the severity of respiratory distress (Silverman score), influence of the therapy on Apgar score, onset of symptomes and haematologic status, necessity of assisted ventilation and neonatal mortality. RESULTS: We found a significant decrease in the incidence of RDS (40%), severity of vital life-threatening distress and neonatal mortality (60%). There were no significant side effects for the babies treated versus untreated. CONCLUSIONS: Antenatal steroids given to the mother in imminence of premature birth are for real benefit in reducing neonatal mortality, increasing the number of survivors among small prematures and improving their quality of life. Even life threatening distress from other causes seem to be less severe, and even an incomplete course therapy conduces to decrease of distress severity. PMID- 12092185 TI - [Metabolic effects of hypolipemic drugs on aged type 2 diabetes]. AB - There are multiple lipids anomalies on diabetes. IDDM has, because of insulin lack, increased levels of triglycerides and afferent lipoproteins. NIDDM, especially obese one, linked by insulinoresistance and hyperinsulinemia, has different and complex anomalies by quality and quantity. There is a specific shape for this anomalies named "B phenotype" with high cardiovascular risk: rise LDL-chol charged with TG and low level of HDL-chol. We searched lipoproteins levels and the effects of simvastatin on aged persons (after 60 years). We randomised 158 cases with obese type II diabetes on a case control study. We concluded that only 28% had high TG levels and 71.8% had low levels of HDL-chol. For HDL-chol this percent is higher over 60 years old group (88.75%) (p < 0.001). Cholesterol has no significant high levels (28.55%) (p < 0.5), and aged group has almost normal levels of cholesterol and triglycerides (p < 0.0001). We administered simvastatin (Zocor) on 86% cases, therapeutically doses, during a period of 6 months to one year. Making lipidograms initially, after 6 months and a year, we proved good effects of Zocor, on lipoproteins levels: rise levels of HDL-chol (p < 0.005), moderate effect on LDL-chol (p < 0.01). At the same time the treatment improving the glucose tolerability to both groups (p < 0.002). PMID- 12092187 TI - [Efficiency criteria for endoscopic diagnostic in upper gastrointestinal bleeding]. AB - Endoscopy plays a major role in the evaluation of upper digestive haemorrhage. We evaluate the difficulties in the introduction of emergency endoscopy in a period of transition, regarding mentalities and errors generated by the learning curve. The study includes 512 consecutive patients admitted with upper gastrointestinal haemorrhage. We retrospectively evaluate some proposed efficiency criteria for the diagnostic endoscopy: the frequency of incomplete endoscopic evaluation (14%), frequency and causes of endoscopic diagnostic errors (3%), frequency of upper digestive haemorrhages with no detectable source (24.2%) and the indication of multiple endoscopic examination. We discuss the causes responsible for the generation of such problems as well as the dynamics of those during the evaluated period. PMID- 12092188 TI - [Endocrinological disorders in association with alopecia areata-a 27 patients study]. AB - Alopecia areata is a dermatological disease, characterized by the loss of hair, which affect men, women and children and can evaluate alone or in association with a variety of other disorders. Between these endocrinological diseases, especial thyroid disorders, have a high incidence. Twenty-seven patients with alopecia areata (12 women and 15 men) aged between 3 and 46 years were endocrinologically investigated. Eighteen of them (66.6%) had endocrinological disorders. Thyroid diseases were present in 10 cases (37%): 4 cases with endemic goiter, 2 cases with nodular goiter and 4 cases with hypothyroidism (1 case with autoimmune thyroiditis, 1 case with nodular goiter, 1 case with cystic goiter and 1 case with hypothyroidism post thyroidectomy for thyroidal lymphoma). Twelve cases (44.4%) were found with tetania. The incidence of thyroid diseases in alopecia areata is higher then in general population (2%), as well as the incidence of tetania. These evidences suggest that it is necessary to make a screening of endocrinological disorders in patients with alopecia areata. PMID- 12092189 TI - [Clinical and biological study of acute diarrhea with mixed etiology in 48 patients]. AB - OBJECTIVES: The study of incidence, clinical manifestation and treatment of acute diarrhea with mixed etiology. MATERIAL AND METHOD: Study of 48 patients with acute diarrhea with mixed etiology admitted in the Hospital of Infectious Diseases of Iasi during 1995-1998. RESULTS: 12 cases (24%) were mixed bacterial infections with the following microorganisms associations: Salmonella + Shigella (10 cazuri); Salmonella + Yersinia enterocolitica (1 case); Salmonella + Rotavirus (1 case). 16 cases (44%) had mixed digestive infections with parasites, in double or triple associations: Giardia intestinalis + Ascaris lumbricoides (10 cases); Giardia intestinalis + Ascaris lumbricoides + Entamoeba coli (1 case); Giardia intestinalis + Enterobius vermicularis (3 cases); Ascaris lumbricoides + Trichiuris trichiura (2 cases). The rest of 20 cases presented mixed infections with bacteria and parasites: Salmonella + Rotavirus + Giardia (2 cases), Salmonella + Shigella + Giardia intestinalis or Ascaris lumbricoides (6 cases), Salmonella + Giardia intestinalis (8 cases); Salmonella + Entamoeba coli (3 cases); Shigella + Trichiura trichiuris + Entamoeba coli (1 case). The majority was male patients from rural areas with age between 5 month and 56 years, the majority being children, 4 cases were found in immunosupressed patients. The clinical symptomatology was dominated by diarrheal syndrome (100%) and the diagnosis was established by clinical characters and confirmed by coproculture and parasitologic exam. The ethiological therapy was guided by antibiogram, in the majority of cases we used fluorochinolones (associated with ceftriaxone in severe cases), together with antiparasitic medications. CONCLUSIONS: In this study predominated the bacterial and parasitic infections, most frequently being isolated Salmonella, Shigella and Giardia intestinalis; the therapy associated fluorochinolones with antiparasitic medication. PMID- 12092190 TI - [Epidemiological, clinical and urographic aspects on a historical sample of patients Wilms tumor]. AB - While the benign renal tumors are exceptional at children, the malign tumors are relatively frequent and they represent the great majority of abdominal tumors on childhood age. In our country, the nephroblastoma is the second in the rate of frequency, after the lymphoma. The radiological investigations play an important role when establishing the diagnostic of Wilms tumor. Although the echography unquestionably offers a series of advantages, most of the authors agree that it can not substitute the intravenous radiography. We subscribe to this opinion, drawing the attention upon the importance of knowing the polymorf urographical aspects which could be determined by development of a Wilms tumor, offering the guarantee of a right interpretation. PMID- 12092191 TI - [Medical and legal involvement of the antisocial aggressive behaviour of the under-age person]. AB - The aggressive behaviour of the child, both the physical and the verbal one, appears due to the conflicts between the environments in which the child is incorporated, but also due to the difficulty of accommodation in a new one. The aggressive behaviour of the child could be a part of the normal way of the child development (for obtain the respect of the group or the autonomy in front of his parents or other persons), but also could be one of the elements, which denote the personality of the juvenile delinquent. The factors which are associated with the aggressive behaviour are: the disorganized family, with many children, the poor family, the alcoholism or drug-dependence of the parents, penal acts of the family members, but also factors which depends on the child: the age between 15 and 17 years old, male, sex head injuries, mental diseases. The facts produced by them could be: theft, burglary, robbery, arson, hitting, rape, sexual perversity, maiming, infanticide, murder. PMID- 12092192 TI - [Mass media involvement in education and information about HIV/AIDS]. AB - AIDS is the first international mass media disease. Throughout the world, most people have learned more about AIDS from radio, television and the press than from personal contacts with health professionals. Mass media AIDS materials influence behavior most when they are designed and developed for different segments of the audience, with the specific needs and concerns of those segments in mind. In the past mass media evoked fear, anxiety and other negative emotional responses to AIDS and didn't proposed practical and realistic ways of action. The journalists made frequently distinctions between innocent victims of HIV, that are infants and recipients of infected blood or blood products through health care, and other "guilty" infected people such as gays, sex workers or african people. Mass media can change behavior and if journalists accept their responsibility in presenting the news, the publicity about AIDS may have a great and positive impact in HIV/AIDS. PMID- 12092193 TI - [Correlations between occlusal dysfunctions and temporomandibular joint disorders]. AB - The occlusal problems and the temporomandibular disorders are the most frequent aspects which we often find to the patients with the dysfunctional syndrome of the stomatognathic system. This study tries to establish the correlation between these two kinds of dysfunctions and to find (if it exists) a direct proportional relation between them. For this purpose we studied 8 cases who had been diagnosed as having the dysfunctional syndrome of the stomatognathic system on the base of the clinical symptoms, tomographs and CT. The results of the study proved that an occlusal dysfunction can affect the temporomandibular joint and vice versa, because all the component parts of the stomatognathic system are in a close relationship and they influence each other, but these results didn't indicate a direct proportional relation between these two kinds of dysfunctions. PMID- 12092194 TI - [Overlay prosthetic solution in subtotal edentation treatment]. AB - The preservation of the natural dental roots represents a big advantage for the overdenture prosthetic appliances. The realization of an overdenture needs a perfect correlation of all the clinical and technological factors, in all the prosthetics steps. Another important aspect is represented by the possibility of a correct treatment of the remaining roots. The remaining roots will be correctly endodontically treated. The root section will be prepared with two inclines bucally and orally located. A metallic cup will cover the root section. On the inner surface of the overdenture the dental technician will realize specific preparations corresponding to this cups. This kind of prosthetic treatment is indicated only for patients with a very good oral hygiene and a good general health. PMID- 12092195 TI - [Magnetic liquid influence upon some plant species of pharmaceutical interest]. AB - It was accomplished a study on the influence of a petroleum magnetic liquid upon two plant species of pharmaceutical interest: Papaver somniferum L. and Chelidonium majus L. Experimental observation aimed: callus accumulation, seed germination, mitotic index and fluorescence of the photosynthesis pigments. The plant samples were taken from in vitro cultures obtained from different explant types while the magnetic liquid was added in the culture media in low concentrations (ml/l). The germination test showed a positive influence of the magnetic liquid, the cell division test revealed an increased mitotic index, callus accumulation rate is enhanced while the fluorescence spectra showed maxima shift for the samples in comparison to the controls. PMID- 12092196 TI - [Case of cystic metastasis of the liver]. AB - The report is about a patient clinic of Iassy from oncologic clinic. He had pain in the right, back and inferior hypocondrium, with irradiation, vomitings, pain less (7 kg in 2 months), weekness. The echography shows three years ago, hepatic cysts (the patient refused the admitted in hospital). Computed tomographic (CT) exam made in jan. 1996 shows three cystic hepatic tumors (2 in right and 1 in left lobes). The CT function revealed sterile biliohematic liquid. The laparotomy made the sharp diagnosis--cephalic pancreatic tumor with cystic metastatic hepatic tumors. The surgical treatment was partial resection of pancreatic tumor and drainage of hepatic cysts, with good postoperator evolution. The intraoperative histological exam found the same kind of malignant tumor in the pancreas and liver. We present the case for his rarity. PMID- 12092197 TI - [Rare location of a hydatid cyst- in the upper mediastinum migrating into the spinal channel]. AB - The authors report a rare and difficult case of hydatic cyst. The case was rare due to the location of the hydatic disease in the upper mediastinum and because of the onset of the disease with a complication: a spinal cord compression syndrome produced by the migration of the hydatic cyst into the spinal channel. The case was difficult because of the diagnostic problems--especially to establish the topography of the lesion and surgical management. PMID- 12092198 TI - [Cutaneous dissemination of visceral neoplasms in women -reported to a clinical situation]. AB - The subject of this case report is a 65 years old female hospitalized for firm, raised, pseudoinflammatory and pseudoangiomatous painless nodular lesions, confluent in large plaques that were localized on the anterior, posterior and left sides of the neck, the anterior and postero-superior areas of the left thorax. These lesions developed in a 3 months period. Histopathological examination of the skin bioptical specimen revealed morphological aspects of an ovarian serous cystadeno-carcinoma metastasis. PMID- 12092199 TI - [Radiological findings in distal radius fractures]. AB - X-ray examination in distal radius fractures is very important for the management and prognostic of these fractures. Anterior and lateral radiographs must be taken in standardised position to avoid errors in measurements. On antero-posterior x ray of the distal radius the important parameters are: radial length (N = 11-12 mm), radial angle (N = 22 degrees-25 degrees) and radial width. For the radial length three methods of measurements (Gartland, Gelberman and Palmer) are presented. On the lateral x-ray the palmar inclination of the distal radius may be measured (N = 10 degrees-12 degrees). Modifications of these parameters associated with distal radius fractures are presented. PMID- 12092200 TI - [Moire effect. Application in medicine and dental technology]. AB - Moire effect is an optical phenomenon that occurs when two similar arrays of alternately opaque and transparent lines or dots are superimposed upon each other. Superimposition of two such similar, but slightly different, line arrays causes interference between the arrays; the interference results in the formation of moire fringes. There is a definite relationship between the pitches of the two arrays and the spacing of the fringes. Therefore, given the pitch of one of the arrays and the distance between the moire fringes, the pitch of the other array can be calculated. The moire effect can be used to determine the development of cutaneous tumors in a very short time-frame and to assess the measurement of dental eruption within 20 minutes. The 3-D facial, palatal and dental morphology can thus be evaluated very accurately. The CAD/CAM dental technology is actually possible due to the moire effect. PMID- 12092201 TI - [The 83rd Congress of the Morphologic Association and the 12th European Congress of Anatomy]. PMID- 12092203 TI - The 2nd Mediterranean meeting "New perspectives in drugs controlled release", 28 29 April 2001, Athens. PMID- 12092202 TI - [The Third Conference of the Romanian Society of Pharmacology, Therapy and Clinical Toxicology]. PMID- 12092204 TI - ["Broncho-pulmonary cancer: progress and current trends" symposium. Iasi, May 31 June 1, 2001. A national therapeutic consensus]. PMID- 12092205 TI - [Anti tobacco consulting]. PMID- 12092207 TI - [Doctor Arnold Stocker , the first author in psychoanalysis in Romania]. PMID- 12092206 TI - [Professor Nicolae C. Paulescu- the true discoverer of insulin]. PMID- 12092208 TI - [A historic editorial published in the world most prestigious medical journals]. PMID- 12092209 TI - Ethics, research, and Helsinki declaration. PMID- 12092210 TI - [Nosocomial infections- major risk in medical procedures]. PMID- 12092211 TI - [From molecular genetics to functional genomic and physiome]. AB - After a brief overview on the advances in the molecular genetics knowledge the paper present new concepts of functional genomics and physiom. First, the results of a group of cell and molecular biology researchers in the filed of genic expression of biologic active endogenous substances, ion receptors and channels are discussed, underlining their influence on actual genomic trend of physiological sciences. Discussions continues on theoretical basis of gene therapy and in the end are presented the researches that made the transition from functional genom to the integrative concepts of physiom, cardiom and proteom, fulfilling the content of the classical notion of normal and pathologic physiology. PMID- 12092212 TI - [18F FDG PET- applications in oncology]. AB - The greatest branch of evolution in Nuclear Medicine that appear in the last period of time, also in medical practice, is scintigraphy with 18F FDG-PET. At the beginning, the images with 18F FDG were used especially in the study of functional cerebral metabolism. Now, new data show that the method presents en major role in cancer diagnosis, localize the tumour and metastasis, and monitoring the treatment in oncology because of the peculiars changes in glucose metabolism of the cancer cells. In this two parts of reference synthesis our goal was to present the following subjects: I) first part include the particularities of glucose metabolism of tumour cells, functionality and components of PET, radiopharmaceutical used in PET, the synthesis of 18F FDG, the clinical procedure and the detection of unknown cancers; II) the second part is about the mains types of tumours identified with 18F FDG-PET (lung, breast, lymphoma, melanoma, colorectal cancer and others). The clinical cases that illustrate the applications are investigated at CERMEP, Lyon, France. PMID- 12092213 TI - Pancreatic enzymes replacement therapy in chronic pancreatitis: an update. AB - Chronic pancreatitis is the major cause of exocrine pancreatic insufficiency which should be compensated by pancreatic enzyme replacement therapy. There are now available a great number of pancreatic enzyme preparations, but encapsulated enteric-coated microspheres and minimicrospheres are considered the enzyme treatment of choice. However, full compensation of pancreatic exocrine insufficiency with enzyme replacement therapy cannot be obtained in all patients with chronic pancreatitis. PMID- 12092214 TI - [The cardiovascular risk after renal transplantation (1): Hypertension and left ventricular hypertrophy]. AB - Cardiovascular disease is the main cause of death in end-stage renal failure treated by hemodialysis or peritoneal dialysis. Though reduced in renal transplant recipients compared to the dialysis population, an excess cardiovascular mortality is still present after transplantation. The authors are reviewing the main data on mortality in the renal transplant population, focusing on major risk factors: hypertension, left ventricular hypertrophy. Chronic immunosuppression is also discussed in this context, as a major determinant of blood pressure elevation after renal transplantation. The presence of these factors and the extent of cardiac and vascular abnormalities in the dialytic patient are closely related to outcomes in the post-transplant period. It is thus mandatory to approach and minimize all these in the dialytic and even predialytic period of chronic renal failure in order to reduce renal transplant mortality in patients with functioning grafts. PMID- 12092215 TI - [Normal and pathologic implication of cytokines]. AB - Cytokines involved in the immune process proof, also, the neurotransmitter role, being synthesized by various cells of the body, with specific genes participation. Cytokines can act on the some membrane receptor like the neurotransmitters, and they can produce diverse biological effects. Pro inflammatory, anti-inflammatory or haematopoetic cytokines as those in the central nervous system can be identified by immunoassay and bioassay methods. Recent works show that the cytokines implication in hypothalamo-hypophyso corticopsuprarenal and gonad axis have consequence on ACTH, corticoids and LH secretion, explaining the ovulatory and gestational disorders. Excessive or insufficient cytokines synthesis, at neuronal level (astrocytes, microglies), explains psychiatric disorders such as schizophrenia, depression, Alzheimer's disease; cytokines (interferon) long term use for the treatment of some diseases, produce irritability, anxiety, delirium, and confusion etc. PMID- 12092216 TI - [Physiological and pathological roles of apoptosis]. AB - An understanding of the cellular events leading to apoptosis is important for the design of new chemotherapeutic agents directed against the types of leukemias and lymphomas that are resistant to currently used chemotherapeutic protocols. Malignant cell proliferation and accumulation depend on an imbalance between the rate of cell production and the rate of cell death. Apoptosis has been demonstrated to be involved not only in the regulation of normal tissues but also in the development and control of tumour cells. The relationship between oncogenes, apoptosis and growth factors is very important in determining prognosis when analysing the potential benefits of different treatment regimens. PMID- 12092217 TI - [The renin-angiotensin system:implications, pathology, therapeutic possibilities, perspectives]. AB - The renin-angiotensin system is initiate by numerous pathological situations which release the renal ischemia: heart failure, arterial hypertension, renal pathology with or without diabetes mellitus. Therapeutic possibilities in renin angiotensin system control are offered by angiotensin-converting enzyme inhibitors, angiotensin II type-1 receptors antagonists, angiotensin converting enzyme inhibitors and neutral endopeptidase inhibitors and angiotensin II type 2 receptors agonists. PMID- 12092218 TI - The role of heat shock proteins in reproduction. AB - Heat shock proteins (hsp) were first discovered in cells exposed to elevated temperature. They are preferentially expressed in response to an array of insults, including hyperthermia, free oxygen radicals, heavy metals, inflammation and infection. In addition hsp are involved in several processes essential for cellular function under physiological conditions. Heat shock proteins are expressed during oogenesis and spermatogenesis. In the early stages of the pregnancy they are expressed both in embryo and maternal decidua. Antibodies to bacterial and human hsp are present at high titers in serum of many patients undergoing in vitro fertilization. Several studies provided evidence that immune sensitization to hsp is associated with unsuccessful embryo development and implantation failure in in-vitro-fertilization (IVF) patients. PMID- 12092219 TI - [White-Bland-Garland syndrome: the maximum natural development of the coronary anastomosis(II)]. AB - The anomalous origin of the left coronary artery from pulmonary artery (White Bland-Garland syndrome) is a very rare coronary malformation, having despite the great mortality in the early childhood, an adult form, characterized by minor symptoms and long course. This paradoxical situation is totally dependent to the complete development of the coronary anastomosis, allowing the right coronary flow to perfuse the left myocardium. The theory of the "terminal arterial perfusion" of the heart, still persistent in some monographs is meeting in this natural situation its strongest opponent argumentation. The coronary anastomosis, clearly visible by coronarography are large enough (in late stages) to inverse the flow in left coronary artery, filling the pulmonary artery. Is the coronary pulmonary shunt, responsible for the majority of symptoms, in the 5th or 6th decades. The adult form of the White-Bland-Garland syndrome is indicative for the maximum result we may hope to reach using the new revascularisation method for ischemic heart disease: new vessels formation and enhancing the coronary anastomosis by growth factors administration therapy. PMID- 12092220 TI - [Spontaneous bacterial peritonitis- undervalued complication in liver cirrhosis]. AB - This complication described rather late and theoretically and practically under evaluated occurs in long term evolution of ascitic hepatic cirrhosis. The positive diagnosis is indicated by the following elements: more than 250 polymorphonuclear count in ascitic fluid, under 1 g/dl protein concentration ascitic fluid, a positive culture for a unique bacterium. Treatment of SBP is made with third generation cephalosporins with immediate favorable evolution. Antibiotic prophylaxis appears to be effective in the prevention of the first SBP episode and long term norfloxacin administration appeared to be very effective in the prevention of the recurrence. The survivors of SBP episode should be recommended for a potential liver transplant. PMID- 12092221 TI - [Small renal cell carcinoma- an update and management]. AB - The detection of small renal cell carcinomas has increased after the introduction of ultrasonography (US) and computed tomography (CT) in the current practice. Their early detection has been extremely important for therapy and prognosis but has opened new diagnostic problems including differential diagnosis, surgical or non-surgical management and imaging follow-up. Small renal carcinomas can be either solitary or multiple, synchronous or metachronous. A synthetic review of incidence, clinical findings, imaging modalities and differential diagnosis of the small renal cell carcinoma will be presented. PMID- 12092223 TI - [Assessment of Tc-99m MIBI uptake mechanism in Mycobacterium tuberculosis and normal cells (fibroblasts and myocytes) cultures]. AB - One of the factors which can explain the scintigraphic Tc-99m MIBI positives images in pulmonary tuberculosis could be the Mycobacterium tuberculosis radiotracer uptake; this can be investigated in vitro, on cell culture, in comparison with normal cell types known to have high (myocites) or low (fibroblasts) uptake. Mycobacterium tuberculosis cultures were realised on Lowenstein Jensen medium, by standard methodology. Myocites and fibroblasts cultures were realized from neonatal rat hearts. Monolayer cells culture were incubated with a same 1.85 kBq/?l Tc-99m MIBI concentration, at 37 degrees C for 15, 60 and 90 minutes. The kinetic was stopped by rapidly washing the cells, with a 4 degrees C physiological saline solution and than scrapped cells were counted for the uptaken radioactivity. The results show that radiotracer cellular uptake (reported at the protein concentration) in myocites was maximum at 60 minutes. Cellular uptake in fibroblasts was very low at all the intervals. In Mycobacterium tuberculosis a peak was observed after 15 minutes, the uptake being similar to that of 60 minutes incubated myocites, considered 100%. After 90 minutes the Mycobacterium tuberculosis uptake was smaller than the 15 minutes value (65.82%). In conclusion, Mycobacterium tuberculosis in vitro uptake results could explain the more positives scintigraphic images in BK positive patients obtain at 15 minutes, in comparison with the delayed images. PMID- 12092222 TI - [Communication and iatrogenesis through communication]. AB - Human intercommunication means transfer of information through many channels, with codes that have biological and socio-cultural determinism. The biological determinism is identifiable through the development of the phonetics and ergomotricity functions, through the possibility of perception of the sensorial organs. The channels of human intercommunication can be verbal, linguistic (the most specialised channel), and nonverbal, extra linguistic channels (mimics, written model, drawing, walking), that are effectively implicated in defining a direct dialogue between the doctor and the patient, completing and emphasising the communication. PMID- 12092224 TI - Contractile effects of intracellularly administered angiotensin II are partially dependent on membrane receptors internalization in isolated rat aorta. AB - In the present study we used the isolated rat aorta as a model to characterize the modulation of contractile effects of extra- and intracellularly administered angiotensin II by dithiothreitol (DTT) and hyperosmotic sucrose. DTT inactivation of AT1 receptor as well as disruption of the clathrin-coated pits by hyperosmotic sucrose significantly inhibited the contraction induced by intracellularly administered AII. We suggest that these intracellular effects of angiotensin peptides are associated with AT1 receptor activation/internalization and may thus be part of the mechanism of angiotensin peptides direct contractile effects in the vascular smooth muscle. PMID- 12092225 TI - [Phenytoin influence on human metanephros]. AB - The influence of Phenytoin on the human embryo is knowing to produce the fetal hydantoin syndrome. Even this syndrome is knowing to produce microcephaly, mental retardation, eyelid ptosis etc it is not mentioned the influence of Phenytoin on the human metanephros. This is the reason of our study, made on human embryos. Their mothers received during pregnancy Phenytoin. In one studied embryo we have remarked a malformation in the metanephros. Even we can not make a strong correlation between the malformation and the Phenytoin we consider that pregnant women should not use Phenytoin during pregnancy. PMID- 12092227 TI - [Correlations between the type of distribution of the branches of the inferior mesenteric artery and the aspect of the mesocolon]. AB - Emergence pattern of inferior mesenteric artery (IMA) branches leaded to description of 6 types of arterial distribution, each of them being correlated with a certain mesocolon aspect. The height of mesosigmoid depends on coalescence pattern and grade, and has a particular importance in terminal colic and retroperitoneal viscera's surgery. For each of the 6 types of distribution are described different ways of differential ligatures, for elongation and descending the colon, initially establishing the predominant collateral trunks. For practicing these ligatures we have to take count of preoperative paraclinic investigations, general and particular haemodynamic conditions and some intraoperatory parameters. PMID- 12092226 TI - [Anatomic and physiopathologic basis for the diagnosis and treatment of intestinal ischemia]. AB - Changes of circulatory parameters in splanchnic territory lead to gut ischemia in an acute or chronic form (postprandial abdominal stroke). The implicated factors in intestinal ischemia are the timing and kind of onset, countercurrent vascularization and the eventuality of anastomotic subsystems for becoming hypertrophic. Superior mesenteric artery (SMA) is the "key" blood vessel implicated in the production of chronic intestinal ischemia, the atherosclerotic lesion being localized more frequent near SMA's ostium or even at ostium itself. Nonoclusif ischemia is induced by another stimulus that provokes mesenteric vasoconstriction, status that affects more the antimesenteric intestinal border, with a larger expansion at mucosa level. The most reseed diagnostic methods are selective angiographies and ultrasound examination. Usually chronic intestinal ischemia is clinically asymptomatic and becomes clinical evident just in the presence of an acute superposed factor or when at least two digestive arteries are affected. Revascularization methods are different, based on the type, mechanism and site of obstruction, each with its advantages and disadvantages. The most used method is retrograde aortomesenteric by-pass, at infrarenal aortic level (that is more accessible than the supraceliac aorta) and SMA reimplantation. PMID- 12092228 TI - [A pilot study about implementation in Romanian medical activity of diagnosis related groups classification]. AB - The inpatient and outpatient classification in homogeneous groups (Diagnosis Related Groups--DRGs and Ambulatory Patient Groups--APGs) it is usually used for the medical resources' management and in second place for the structure's and the sanitary system's function evaluation, for the establishment of necessities and priorities in medical care of a population, for the establishment of the used resources etc. These elements given by the method are essential for a sanitary reform. In this whey the possibility of implementation for the method in Romanian medical practice, we've proposed to evaluate it in a pilot study. METHODS: The study was made on an adult population sample from Cluj-Napoca. It was made out of 311 subjects. The data were taken in unitary mode using a record questionnaire. The database was performed in the ACCESS program. For the statistic there were used the information pack EpiInfo version 6, 04. 1996. The reference year was 1997. To observe the evolution of the phenomenon we compared with a control group from 1987 composed by matching 1:1. RESULTS: In the sample females were significatively dominant (65.4%, p < 0.00001) and elders (the age group of 60-69 years represent 54%, p < 0.00001). In 1987 the elders > or = 60 years were the once how had in a significantly way more often medical care both in and outpatient. In 1997, the medical care for adults was close to the once for elders. In evolution, for a 10 year period, the ambulatory assistance decreased and the specialized assistants in the hospital increased (unique and multiple). The risk of hospitalization in 1997 compared to 1987, is significatively bigger: OR = 2.56 (IC 95%: 1.61-4.09), p < 0.001. The pathology assisted in ambulatory and in hospital was represented more often by the cardiovascular system affections (CMD 05-61.1%) in witch the arterial hypertension is dominant (DRG 191). CONCLUSIONS: To produce DRGs and APGs, in the actual sanitary system, from the university center Cluj-Napoca it is possible, but the development of the actual informational system is necessary. The implementation of the classification method needs from the medical practice side it's standardization because the homogeneous patients consume the same quantity of resources from the medical point of view. The medical service separation in smaller and bigger hospitalization is necessary. PMID- 12092229 TI - [Clinical aspects of the left ventricular diastolic dysfunction at patients with coronary heart disease]. AB - Is is known that, besides the systolic disfunction, the presence of diastolic disfunction influences the prognosis of patients with coronary heart disease. The aim of the paper is to study the influence of the presence of diastolic disfunction upon the clinical profile and the functional capacity of the patients with coronary heart disease. METHODS: 189 patients with ischemic heart disease (chronic myocardial infarction, stable angina, ischemic cardiomiopathy) underwent an echocardiographic examination. The diastolic disfunction was assessed at the Doppler examination, by the the ratio of the E and A velocities. Consequently, 138 patients had a E/A ratio under 1 or above 2 and were considered with diastolic disfunction (group A); the other 47 with E/A ratio between 1-2 were included in group B. After the clinical evaluation, 80 patients underwent an exercise testing (47 of group A and 33 of group B). RESULTS: Group A had a greater mean age (62.8 +/- 11.7 vs 51.5 +/- 11.6 years, p = 0.00006) and more females (72.5% vs 46.8%, p = 0.001); dyspnoea was more frequent and severe (NYHA II-IV), the arterial pressure higher (146/85 +/- 25/13 mmHg vs 136/79 +/- 26/15 mmHg, p = 0.004) and cardiac rate higher (82 +/- 18 vs 69 +/- 11/min, p = 0.0003). The echocardiographic assessment revealed a more severe systolic disfunction and a lower ejection fraction (50.3 +/- 10.6% vs 55.7 +/- 10.6%, p = 0.003). At the exercise testing, the maximum level and the duration of exercise were lower at group A than group B (5.41 +/- 2.07 MET vs 7.23 +/- 2.78 MET, p = 0.001 respectively 8.4 +/- 4.5 min vs 11.4 +/- 4.9 min, p = 0.01). CONCLUSIONS: The presence of diastolic disfunction is higher at older patients and at those with associated arterial hypertension. Dyspnoea correlates with the presence and severity of diastolic disfunction and the exercise capacity is lower at these patients. PMID- 12092230 TI - [Gastroesophageal reflux during pregnancy: 24-hour esophageal ph monitoring]. AB - Gastroesophageal reflux (GER) occurs in 30-50% of all pregnancies. The progressive rise in plasma progesterone has been suggested as a possible mediator of GER during pregnancy. Recent advances in technology have made it possible to detect GER through monitoring of esophageal pH for prolonged periods, including sleep. 24-hour pH monitoring is the proper method for diagnosing GER in pregnant women. If 24-hour esophageal pH monitoring is to be a useful diagnostic tool, it must reliably discriminate GER patients despite daily variations in distal esophageal acid exposure. To address this issue, we studied 62 women (30 healthy non-pregnant women without GER symptoms and 32 pregnant women with GER symptoms heartburn, acid regurgitation) with 24-hour esophageal pH monitoring. Intrasubject reproducibility of three pH parameters to discriminate the presence of abnormal acid reflux was determined (DeMeester score, Kaye score, circadian one hour diagram for pH < 4). Each patient was interviewed, using a reliable questionnaire detailing individual habits, life style characteristics and symptoms, at four time points during the first, second, third trimesters of pregnancy and post-partum period. Symptoms of GER are common in pregnancy and although GER rarely endangers maternal or fetal health, it can significantly affect patient comfort and quality of life. We conclude: 1. GER is almost constantly present during pregnancy, increasing with gestational age. 2. The most important pH--parameter is DeMcester score. 3. Heartburn disappear after delivery. 4. 24-hour esophageal pH monitoring is the gold standard for measuring acid exposure and is a reproducible test for the diagnosis of GER in pregnancy. PMID- 12092231 TI - [The gallstone and ischemic heart disease]. AB - I analyzed IHD and gallstone, in a study on 1122 patients. OBJECTIVES: The relation between IHD and gallstone incidence. METHODS: In this study--516 patients had IHD, with physical examination and Ecg, from 1122 patients with gallstone which were examined abdominal ultrasonography. RESULTS: I found that 45.98% from patients with gallstone had IHD. CONCLUSIONS: IHD can coexist with gallstone and the association between these two is 2.8 times more frequent in women. PMID- 12092232 TI - [Contribution of ultrasonography in the evaluation of subacute thyroiditis]. AB - Ultrasonography is a useful tool in diagnosing and monitoring thyroid pathology. The aim of this study was to evaluate the value of thyroid ultrasonography in the follow-up of subacute thyroiditis (SAT). The study group included 48 subjects with clinical and biological diagnosis of subacute thyroiditis. All patients were evaluated by ultrasonography and followed-up for a mean period of 12 months. The evaluation protocol included biological data (TSH, T4, erythrocyte sedimentation rate--ESR) and ultrasonography, performed at 1, 2, 3, 6, and 12 months. Initially, all patients had an enlarged thyroid volume (median = 30.5 ml, range 23-90) and a low echogenicity, with an inhomogeneous aspect in 54.1% cases. After a mean period of 3 months thyroid volume had a significant reduction (median volume = 20.2 ml, range 7-36, p < 0.0001). Echogenicity was also significantly improved, with a decrement, on a semi-quantitative scale of 4 degrees (1 = normal, 4 = intense hypoechogenicity), from 3.25 to 2.48 (p < 0.0001). Almost half of the patients (45.8%) presented persistent (although slighter) hypoechoic/inhomogeneous pattern. Ultrasonographic abnormalities were not correlated with the intensity of the inflammatory syndrome and/or the thyroid status. Recurrence appeared in 10 (20.8%) patients. All patients presented a new thyroid enlargement (from a median = 16.1 ml to 31.5 ml, p = 0.056) and an extension of hypoechoic regions. The risk of recurrence could not be correlated with thyroid function, inflammatory syndrome or ultrasonographic aspect. There were not significant differences between the subgroups of patients with and without recurrence concerning the initial thyroid volume (p = 0.889), echogenicity (p = 0.735), TSH (p = 0.321) or ESR (p = 0.1332). Thyroid ultrasonography is useful not only for the initial diagnosis but also for the follow-up of patients with subacute thyroiditis. Remission and recurrence can be appreciated and monitored by the ultrasonographic pattern of the thyroid. PMID- 12092233 TI - [Nebivolol treatment in essential arterial hypertension]. AB - Nebivolol is a lipophilic beta 1-blocker. It is devoid of intrinsic sympathomimetic or membrane stabilising activity but appears to have nitric oxide mediated vasodilatory effects. Nebivolol 5 mg once daily is well tolerated in patients with hypertension. Adverse events are infrequent, transient and mild to moderate. Those reported most often include headache, fatigue, paraesthesias and dizziness. Several studies reported no signs of orthostatic hypotension with Nebivolol. PMID- 12092234 TI - [Treatment of alopecia areata with diphencyprone]. AB - Topic immunotherapy in alopecia areata consist in a mild contact dermatitis with help of a chemical substances, with a high potency of sensitization, such as dinytroclorbenzenul (DNCB), squaric acid dibutylester (SADBE) and diphencypronil (DPCP). Eight patients with alopecia areata, 3 with mild form and 5 with severe form was treated with DPCP in acetone solution. We have obtained a positive result in 3 cases (37.5%), one with total regrows and 2 with partial regrows and a negative result in 5 cases, 3 with partial regrows and loss of the hair in other areas and 2 cases with no response after 24 weeks. We consider this method like an alternative therapy in severe alopecia areata resistant at other treatments. PMID- 12092235 TI - [Displaced femoral neck fractures: internal fixation or arthroplasty?]. AB - Femoral neck fractures are a challenge for orthopedic surgeon due to the high rate of complication for such fractures. The aim of this study was to propose a treatment algorithm based on author's current concept in management of femoral neck fractures. Fractures were classified according Garden in four types but, for practical reasons, fractures were considered undisplaced (Garden I and II) and displaced (Garden III and IV). Treatment method was chosen based on next criteria: patient's age, associated pathology, patient's mental status and expectations, quality of the bone (osteoporosis), type and age of fracture, quality of reduction. Based on this criteria, the proposal algorithm may led to close reduction and internal fixation, open reduction and internal fixation or arthroplasty. PMID- 12092236 TI - [Factors which may influence the surgical outcome in myasthenia gravis]. AB - OBJECTIVE: This study intends to assess the factors which may influence the surgical outcome in myasthenia gravis. METHOD: 52 myasthenic patients were operated on in the IIIrd surgical unit, Iasi, undergoing extended thymectomy through a complete longitudinal sternotomy during a period of 21 years (1980 2001), the surgical outcome being variable. The following factors: age, clinical stage (Osserman classification), the history length and the histopathology of the specimen were analysed in respect with the surgical outcome which varied from complete remission and clinical improvement till no effect and death. Out of 52 patients, 28 were under the age of 30 years and 24 above 30 years of age. 5 patients were operated on in the first clinical stage, 34 in the second clinical stage, 10 in the third stage and 3 in the fourth. 21 patients had had under 1 year history of the disease, 11 patients under 2 years and 20 patients over 2 years. 40 cases presented non-tumoral pathology of the thymus gland and 12 cases presented thymomas. RESULTS: The best surgical results were recorded in patients under 30 years of age, operated in the first and second clinical stage, with a history under 1 year and non-tumoral pathology of the thymus. CONCLUSION: Myasthenia gravis is the only one acquired autoimmune disease in which surgical treatment offers the chance of complete remission if the following criteria are encountered: young patient, short history, initial clinical stage and non-tumoral pathology. PMID- 12092237 TI - Sensitivity and resistance to antibiotics of Streptococcus pneumoniae involved in severe infections (1990-2000). AB - S. pneumoniae implicated in severe infections presents now a high frequency of the resistant strains to Penicillin G or multiresistance in the whole world. Our study on 136 strains of invasive pneumococcus isolated in the last 10 years showed a medium resistance to penicillin of 22.8% with yearly variations. The resistance to Rifampin, Cloramphenicol and Eritromycin was of 4.4%, 7.8% and 8.0%. The treatment of pneumococcal infections must be adapted to actual antibiotypes. These results present therapeutic implications. PMID- 12092238 TI - Spinal epidural abscess. AB - In a retrospective study, 68 patients with Spinal Epidural Abscess (SEA) were reviewed. Of these, 66% had different predisposing factors such as staphylococcal skin infections, surgical procedures, rachicentesis, trauma, spondilodiscitis. Abscess had a lumbar region location in 53% of cases. Staphylococcus aureus was the most frequent etiological agent (81%). The overall rate of mortality in SEA patients was 13.2%. PMID- 12092239 TI - [The evaluation of sound level in dental practice]. AB - Noise pollution is one of the most important problems of the contemporary world. Dentistry is one of the most affected areas because the activity in a dental practice involves the use of different devices that generate noise. The purpose of our study is to evaluate the sound level in a dental practice with 4 dental units using a complex system, which comprises: a Sound Blaster Live 5.1, a Dual microphone, a PC and special software for the acquisition and data analysis. The sound level detected by us is similar with the data from the international literature with some particular aspects and suggests that the dental practice is a noise polluted environment although most of the sound levels are beneath the damaging noise level for the human ear (85 dB). PMID- 12092240 TI - Morphological features in the embryological development of the anterior arch of the mandible. AB - OBJECTIVES: Our study is focused on the typical morphological features of the development of the mandible. There were investigated both specific elements for the ossification process as well as aspects of the developing tooth germs. METHODS: We performed transmission electron microscopy analyses on sections obtained from 15 human embryos aged between 6 and 20 weeks. The sections were acquired from specific areas of the anterior arch of the mandible, corresponding to the sites of development of the tooth germs, namely the incisors and the canines. RESULTS: There were observed some characteristic elements for the intramembranous ossification process (mesenchymal cells, collagen fibers, osteoblasts, bony spicules), and for the different stages of odontogenesis (bud, cap). CONCLUSIONS: The results confirm the important role of this territory and highlight the primary elements of the intramembranous ossification and of the odontogenesis, as essential steps in the development of the head and the face. PMID- 12092241 TI - Zeolitical materials for microbiological filtrations. AB - The paper presents some aspects about synthesis of new materials called zeolites which can be utilised as filters with wide application in clinical practice in microbiology and in pharmaceutical industry. The SAPO-5, SAPO-11 and SAPO-34 zeolites were synthesised by hydrothermal crystallisation and using different mixtures. The crystallisation was carried out in teflon-lined stainless steel autoclaves at 180-195 degrees C for 1-2 days, under autogenous pressure. Filtering properties of these materials are different because their pore diameters are different. PMID- 12092242 TI - [Mushroom poisoning in an autoimmune hepatitis patient]. AB - Mushroom poisoning is responsible for initiation of severe life threatening syndromes, especially hepatic cytolysis. We are describing a case in which the helvelian syndrome induced by mushroom meal was masking the preexistent autoimmune hepatitis type 1. The course of disease under corticoids was satisfactory after one month of surveillance. PMID- 12092243 TI - [Chronic dacryocystitis fistulization in a Rubinstein-Taybi syndrome]. AB - The article presents the treatment difficulties of the chronic dacryocystitis at an eight month old patient, diagnosed with Rubinstein-Taybi syndrome. The difficulties occurred as a result of numerous antibiotic treatments and surgical procedures suffered by the patient since his second week of life. PMID- 12092245 TI - [Compression hip screw in trochanteric fractures. Conclusions based on 128 cases]. AB - Compression hip screw is currently used in our department since 1998 and we operated 128 trochanteric fractures using DHS (Synthes) or Omega system (Howmedica-Stryker). We recommend the use of this implant in all stable fractures, most unstable fractures and we believe it is useless in reverse oblique trochanteric fractures. Best time for surgery seems to be the first or second day after admission. A preoperative planning must precede the surgery. We prefer the lateral approach of the hip with "L" type incision for vastus lateralis. For insertion of implant AO original technique was used. Mechanical complication occur only in unstable fractures but the failure rate was lower than for other implants. Patient with stable fractures were allowed full weight bearing immediately postoperative, this being delayed 6 weeks for patients with unstable fractures. We recommend compression hip screw as a very useful implant in trochanteric fractures. PMID- 12092244 TI - [Fertility in prolactinomas--considerations concerning some clinical cases]. AB - Prolactinoma is a frequent endocrine cause of infertility in both man and women. The aim of this study was to evaluate female fertility in association with hyperprolactinemia. The study is a retrospective one, based on the cases monitored in the Endocrinology Department of Iasi. From the 113 cases of prolactinoma in women we have selected 83 women aged between 18-45 years (fertile age). The diagnosis was suspected on clinical bases: endocrine (amenorrhea galactorrhea, associated or isolated, spaniomenorrhea, infertility), and tumoral (headache, visual disturbances) syndromes and confirmed by biological (PRL dosage) and morphological (pituitary CT) tests. We classified the found associations between prolactinoma and fertility in: no correlation (previous pregnancies with no desire for another child--76 patients = 75.2%); prolactinoma diagnosed immediately after a pregnancy by persistency of galactorrhea, lack of menses reapparition, headache (16 cases = 15.8%); prolactinoma diagnosed before pregnancy, which was obtained thanks to the hyperprolactinemia treatment (9 patients = 8.9%). All 9 patients of the last group were in remission at the moment of pregnancy apparition and had a normal pregnancy, giving birth to normal children. Infertility associated with prolactinoma is reversible with treatment. Lowering of prolactin levels to normal is often necessary to permit ovulation. Bromocriptine used by pregnant women appears to be safe for the developing fetus, at least in our study where the treatment was interrupted in most cases in the first trimester of pregnancy. PMID- 12092246 TI - [Magnetic carrier method- an expectation for antitumoral therapy]. AB - In order to enhance antineoplastic agents efficiency new therapy methods were developed one of which being magnetic targeted chemotherapy (MTC). MTC method consists in a permanent or reversible selective binding of highly magnetic particles (carriers) with drugs, antibodies et. al., followed by targeting or adhesion of these complexes to the tumor using an external magnetic field. At this early stage of targeting tumors with magnetic particles, there are reasons to be optimistic. PMID- 12092247 TI - [Prognostic factors in the primary invasive vaginal carcinoma]. AB - In the period 1965-1988, 125 women with primary invasive vaginal carcinoma were treated with radiotherapy. 53 (42.4%) survived 5-year without evidence of disease. In the Cox multivariate analysis three variables were independently related to survival: age of the patients, grade of differentiation of tumor and the clinical stage of illness. PMID- 12092248 TI - [Identification of encoding and regulatory sequences of RAR-alpha receptor in intraepithelial neoplasia and in invasive cervical carcinomas associated with HPV types 16 and 18]. AB - The purpose of the study was the analysis of regulatory (region 5 gen) and encoding sequences of RAR-alpha receptor in cervical dysplasia and invasive cancer associated with HPV infection. A PCR method with the use of specific primers was applied to amplify and detect DNA sequence of various HPV types. A PCR-SSCP method was used to analyse the sequence of RAR--a receptor. The study revealed changes in structure of regulatory and encoding sequences of the investigated RAR receptor. Utmost changes were found in DNA isolated from tissues with diagnosis of adenocarcinoma colli uteri. PMID- 12092249 TI - [TVT procedure--the critical analysis of clinical effectiveness and complications among first 100 cases]. AB - RATIONALE: Since its clinical debut in 1996 TVT procedure has been offering an excellent clinical tool both for patients and surgeons to treat SUI suffering women. Since the learning curve is an important factor influencing the rate of surgical complications as well as the clinical outcome of any new surgical technique we decided to analyse the first 100 cases of SUI women treated with TVT technique. AIM OF THE STUDY: To analyse clinical effectiveness and complications among first 100 consecutive cases of TVT procedures. MATERIAL AND METHODS: One hundred women aged from 30 to 89 years (old mean 55.6) were hospitalised in our Department from September 1999 to November 2000 because of SUI as confirmed by the complete urogynecologic assessment including history and physical examinations, catheterised residual volume determination, and multichannel urodynamic testing. TVT procedure was performed as originally described by Ulmsten et al. except the fact that 43 operations were performed not under local but epidural anesthesia. For 80 patients TVT was offered as the primary surgery for SUI treatment whereas for 14 it was the second and for 6 the third or more attempt. RESULTS: Operation was performed without any technical difficulties in all cases (mean time 25 min). Bladder injury (treated by 1 or 2 days catheterization) occurred in 9 cases, urinary retention in 13 (in 6 cases after 10-14 days tape was cut). Urinary tract infections occurred in 4 women despite intraoperative 2.0 g i.v. piperacillin prophylactics and fosfomycin trometamol 3.0 g p.o. in first postoperative day. Subsequent urge incontinence developed in 4 patients. Clinical efficacy based on medical history, cough test and Gaudenz questionnaire was 95% (observation period 3 to 16 months, mean 8 months). CONCLUSIONS: In our opinion TVT procedure is an operation of choice to treat SUI in women regardless of their age because of its excellent clinical effectiveness accompanied by its technical simplicity, reproducibility and low percentage of serious complications. Previous urogenital surgery is a risk factor for bladder perforation during TVT procedure. PMID- 12092250 TI - [Usefulness of fetal echocardiography in the prenatal diagnosis of Down syndrome based on material from the Department for Diagnosis of Congenital Malformation's of the Institute "Polish Mother's Memorial Hospital' in Lodz between 1994-1999. Part I]. AB - Between 1994-1999 at the Department for Diagnosis of Congenital Malformations at the PPMH Institute there were ultrasound and echocardiography examinations performed in 40 fetuses with Down Syndrome. In our study most of the cases come from low risk groups of pregnant women. Congenital heart defects (AV-canal in 13/18 cases) were the most common anomalies diagnosed prenatally in fetuses with Down Syndrome. In fetuses with DS, functional cardiac anomalies with normal heart anatomy were recorded: disproportion and hypertrophy in 4/12 cases, tricuspid valve regurgitation, bright spot and pericardial effusion in 3/12 cases. The aim of our study was a retrospective analysis of fetuses with DS, which were diagnosed at the Department for Diagnosis of Congenital at the Institute of "PMMH" in 1994-1999 years. PMID- 12092251 TI - [Usefulness of fetal echocardiography in the prenatal diagnosis of Down syndrome based on material from the Department for Diagnosis of Congenital Malformation's of the Institute "Polish Mother's Memorial Hospital' in Lodz between 1994-1999. Part II]. AB - In the years 1994-1999 at the Department for Diagnosis of Congenital Malformations at the PPMH Institute, there were ultrasound and echocardiography examinations performed in 40 foetuses with Down Syndrome. In our study most of the cases come from low risk groups of pregnant women. Extracardiac malformations were diagnosed prenatally in 25% fetuses with Down Syndrome. The most common anomaly from extracardiac malformations was duodenal atresia (13/40 cases- 32.5%). PMID- 12092252 TI - [Analysis of the course of pregnancy, delivery patterns and the weight of newborns of female smokers]. AB - The authors scrutinized the course of pregnancy, delivery patterns and the weight of newborns among 1741 women who delivered between 1993 and 1998 at I Clinic of Obstetric and Gynecology at Warsaw Medical University while the control group consisted of 674 non smoking women, who delivered over the same period. Spontaneous abortion, preterm delivery and intrauterine grow retardation were all registered more frequently in the study group. Moreover, cases of lower weight in smoking mothers' newborn occurred more often. PMID- 12092253 TI - [The course of neonatal period in neonates of epileptic mothers]. AB - In prospective analysis we compared 211 newborns of women with epilepsy (examined group) and 211 random chose newborns of mothers, without neurological and psychical diseases (control group). Newborns were admitted to ICZMP in 1990-1998 years. In the examined group, the percentage of premature birth, hypotrophia and inborn malformations was higher as compared with the control group. Among biochemical abnormalities the most frequent was hypocalcemia. By means of clinical examination we found circuits of head diminished and most frequently fine tremor in the examined group. Analysis of blood morphology showed lower values of haematocrit, a lower level of haemoglobin and lower numbers of erythrocytes. PMID- 12092254 TI - [The role and clinical meaning of inhibins in female reproductive function]. AB - The existence of inhibins was suggested in 1932, but their complete function and all places of secretion still aren't recognized. Inhibins are glycoproteins produced by the granulosa and theca cells of the ovary and, in lower proportions, by some extragonadal tissues. During pregnancy, placenta becomes the predominant secretory organ for inhibins. Clinically, inhibins may serve as sensitive tumor markers in postmenopausal women, or as useful tools for evaluating ovarian reserve in infertile women. They may also be used in the diagnosis of materno fetal disorders, or be markers of function of the luteal phase. PMID- 12092255 TI - [Clinical aspects of Livial in postmenopausal replacement therapy]. AB - It's a review of literature on the clinical effects of Livial. Livial (generic name-tibolone) makes a therapy of choice for postmenopausal women. It's a synthetic steroid with estrogenic, progestagenic and androgenic properties due to its tissue specific effects. Livial is effective in reducing wasomotors symptoms and has positive effects on mood and libido. It also prevents bone loss and reduces risk of fractures. Livial is safe for cardiovascular system. The incidence of side effects is very low, about 3 in every one hundred. Because of its progestagenic activity in endometrium it results in endometrial atrophy but in vagina the studies showed oestrogen--like effects, i.e. improvement in vaginal dryness. Livial may be used as an "add-back" therapy in conjunction with GnRH agonists treatment. The rates of discontinuation is lower than HTR preparations, about 25%. Livial may have benefits over conventional HRT in older women (over 65 years old), in women with history of endometrial cancer, breast cancer and endometriosis. PMID- 12092256 TI - [Using of Therma-Choice system in treatment of metrorrhagia in case of porphyria]. AB - The authors present the use of Uterine Balloon Therapy in the long term metrorrhagia resistant to conservative treatment, in the case of porphyria. The method was used on a 44 year old female, suffering from acute intermittent liver porphyria for 9 years. PMID- 12092257 TI - [Pregnancy and delivery after conservative treatment of borderline ovarian tumor with FIGO stage III C--a case report]. AB - This study presents a case of pregnancy after conservative surgical treatment of borderline ovarian tumor with III C stage according to FIGO classification. The caesarean section in 37 week of pregnancy was performed and connected with very precise staging. In normal-appearing both ovaries the foci of borderline ovarian tumor were found. There were no foci of borderline ovarian tumor or foci of invasive carcinoma in tissues routinely sampled during the second look operation. All benefits and costs of conservative surgical approaches to very young, childless women with advanced borderline ovarian tumor were exactly estimated. PMID- 12092258 TI - [Effect of recombinant activated factor VII (RFVIIA; NovoSeven) in a patient in haemorrhagic shock after obstetrical hysterectomy]. AB - Perinatal bleeding may lead to a fatal outcome in about 10% of cases. We present a case of a 29 year old woman in childbirth, in whom a preparation of recombinant activated factor VII (rFVIIa, NovoSeven, Novo Nordisk A/S, Bagsvaerd, Denmark) was used with success to control intraoperative bleeding during haemorrhagic shock after caesarean section and obstetrical hysterectomy, in the course of the patient's third abdominal cavity revision for continuing blood loss. Administration of the preparation enabled for quick intraoperative control of haemorrhage, which allowed safe and clear identification and elimination of the cause of bleeding. No significant clotting abnormalities were noted at postoperative monitoring of the coagulation system. rFVIIa seems to be an effective alternative treatment of life-threatening haemorrhages in obstetrics, allowing for quick arrest of bleeding, and thus safe control of intraoperative local hemostasis. PMID- 12092259 TI - [From the history of Radom's gynaecology]. AB - The aim of the work is to present a study of the gynaecology in Radom until 1947. The base of this study are prints, memoires and anniversary studies, both published and in typescript. Considering the scantiness of sources, disintegrated archives and time passed, the study is popularizing in character. PMID- 12092260 TI - [Report on participation in 26th Annual Congress of the International Urogynecological Association, Melbourne, Australia, August 5, 2001]. PMID- 12092261 TI - Successful interdisciplinary documentation through nursing interventions classification. AB - Automated documentation systems enable health care professionals to develop dynamic, interdisciplinary care plans. Use of a standardized nomenclature provides a common framework and language for the communication of the plan, and also can support data collection to determine best clinical practices. A comprehensive patient care record will link the interdisciplinary care plan to clinical charting and the documentation of outcomes. This article reviews efforts to improve interdisciplinary communication by computerizing the documentation system, using the Nursing Interventions Classification (NIC). PMID- 12092262 TI - Patient first: a tri-fold approach to health care transformation. AB - The installation of new technology affords health care facilities the tri-fold opportunity of (1) upgrading processes and systems, (2) launching providers into the forefront of health care, and (3) providing the most advanced quality of care. Redesigning patient care processes to align with new technology further presents opportunities to transform culture, enhance service excellence, increase staff accountability and promote self-esteem. Leveraging each advantage is a significant commitment that requires a comprehensive change strategy. Eastern Maine Medical Center in Bangor, ME serves as a case study for success. PMID- 12092263 TI - Informatics competencies for nurse managers and their staffs. AB - Nurses have always dealt with data, information, and knowledge. Therefore, as automated information systems become increasingly important in health care, all nurses should have core competencies in computer skills and data and information management. These competencies differ according to the skill level of the nurse, from a beginning nurse to nursing manager, and on to those who specialize in nursing informatics. PMID- 12092264 TI - Out with computer literacy, in with information literacy: the nurse manager's challenge for the 21st century. AB - As computer literacy has given way to information literacy, knowing how to manage and utilize the information that technology brings is essential for the nurse manager of the future. This article examines some of the challenges facing nursing as it attempts to turn data into information and usable knowledge. PMID- 12092265 TI - Nurses in corporate America: embracing power through influence. AB - Executive positions in corporate America offer nurse leaders the opportunity to influence product development and services delivered to ensure that the best possible solutions are provided to health care organizations, providers, and patients. This opportunity to "make a difference" is a critical component for nurses' attraction to migrating to the business side of the health care industry. However, making the transition from leadership positions in health care delivery organizations to corporate businesses carries big challenges. A major demand is for nurse leaders to adjust from direct span of control organizational models to matrix management structures used in complex business organizations. PMID- 12092266 TI - Leadership in the digital age: a framework to balance high touch with high tech. AB - The next great age will be influenced heavily by the physical sciences, biological sciences, and nanotechnologies. This article presents case scenarios highlighting the technologic possibilities for the next age. It also presents a potential leadership model for the digital age and beyond. Finally, it provides insight as to how nurse managers can reconcile one of the major dilemmas in nursing--balancing "high touch" with "high tech." PMID- 12092267 TI - Health care, ethics, and information technologies. AB - This essay explores how ethics, computing, and health care intersect in medical informatics. It discusses the power technology places in the hands of health care professionals and the ethical problems they may encounter as a result of that power. PMID- 12092268 TI - Patient safety and I.T.--it's everyone's concern! PMID- 12092269 TI - Information systems enhance efficient and effective nursing practice and management. PMID- 12092270 TI - Finances and decision making. PMID- 12092272 TI - The use of aggregate data for measuring practice improvement. AB - Practice improvements are much needed in health care but are difficult to implement and to measure. Without records that include aggregate data, outcomes cannot be compared. Aggregate data are defined as data not limited to one patient, but data that are tracked across time, across organizations, across patient populations, or across some other variable. There are several issues that complicate the gathering of aggregate data, including data availability, format, management and definition, as well as the processes of trending and comparison to a standard benchmark. PMID- 12092271 TI - What HIPAA means for your clinical practice. AB - The Health Insurance Portability and Accountability Act (HIPAA) defines simplification, privacy, and security standards that have prompted health care providers to embark on a process of formalizing long-standing values to protect patient information. Targeting vulnerable practices, raising awareness through education, and formalizing reasonable policies and procedures will go a long way toward assuring privacy and confidentiality for patients and clients. This article offers solid approaches on how to develop and implement privacy and security measures that will engender patient trust and confidence. PMID- 12092274 TI - Stereotactic radiosurgery failure in acoustic neuroma. PMID- 12092273 TI - Using standard desk-top tools to monitor medical error rates. AB - Many experts believe it is time to do something about the high number of medical errors and adverse events highlighted in the recent IOM report. This article examines how front line managers can use a valuable quality improvement/assurance tool--the statistical process control chart--to track and analyze medical error rates. It includes a step-by-step guide to the creation and interpretation of a demonstration chart, discusses potential barriers to the use of statistical process control charts, and suggests ways to overcome those barriers. PMID- 12092275 TI - Otomycosis. PMID- 12092276 TI - Papilloma of the dorsal aspect of the soft palate: a difficult lesion to visualize. PMID- 12092277 TI - Bilateral granuloma and varicosity in the midportion of the vocal folds. PMID- 12092278 TI - Anomalous course of the cervical carotid arteries. PMID- 12092279 TI - A patient with ear pain who had panic attacks while driving over bridges and in wide-open spaces. PMID- 12092280 TI - Endoscopic Zenker's diverticulotomy. PMID- 12092281 TI - Peritonsillar abscess: a study of 724 cases in Japan. AB - We reviewed the records of 724 patients diagnosed with peritonsillar abscess who had been admitted to our hospital between January 1988 and December 1999. We analyzed their clinical features, disease course, and treatment. The male:female ratio was 3:1, and approximately two-thirds of these patients were between 20 and 39 years of age. The most common aerobic bacteria cultured from patients' pus were alpha-hemolytic streptococci. Severe complications---including deep neck infections and mediastinitis--were seen in 13 patients (1.8%). This complication rate suggests that patients with peritonsillar abscess should undergo immediate incision and drainage rather than needle aspiration. PMID- 12092282 TI - The Hunsaker Mon-Jet ventilation tube for microlaryngeal surgery: optimal laryngeal exposure. AB - Methods of delivering and monitoring anesthesia during microlaryngeal surgery are constantly evolving. In 1994, Hunsaker and colleagues introduced a laser-safe subglottic Mon-Jet ventilation tube, which has the ability to periodically measure end-tidal carbon dioxide levels. We conducted a retrospective review of 84 consecutive patients who had undergone microlaryngeal procedures with the aid of the Hunsaker Mon-Jet tube. Study parameters included the length of anesthetic induction and recovery times, the duration of surgery, the degree of surgical access to the larynx, and the incidence of anesthetic and surgical complications. We found that anesthetic induction and recovery times with the use of the Mon-Jet tube were comparable to those seen with standard endotracheal intubation. We also observed an apparent reduction in surgical time and a consistent subjective improvement in surgical visualization and access. The complication rate was acceptable, airway control was adequate, and use of the Mon-Jet tube was safe in all patients. We conclude that the Mon-Jet tube is a safe and effective subglottic jet ventilation system and that it has distinct advantages over other methods for both the surgeon and the anesthesiologist. PMID- 12092283 TI - Primary hyperparathyroidism secondary to simultaneous bilateral parathyroid carcinoma. AB - Parathyroid carcinoma is a rare malignancy, and experience with its management is limited. It is generally accepted that surgery is the treatment of choice, and the initial operative intervention might represent the only chance for long-term success. At the time of surgery, a high index of suspicion for carcinoma is vital to performing the appropriate surgical procedures, because frozen-section diagnosis of parathyroid carcinoma is notoriously unreliable. In this article, we describe our experience with this rare entity. PMID- 12092284 TI - Early-onset postirradiation sarcoma of the head and neck: report of three cases. AB - Postirradiation sarcomas of the head and neck are rare. When they do occur, most appear at least 10 years following radiation therapy. We report three cases of early-onset (1, 2, and 7 yr) postirradiation sarcoma. Physicians who care for previously irradiated patients should consider the possibility of a postirradiation sarcoma whenever they see a suspicious lesion, regardless of the amount of time that has passed since radiation therapy was administered. The original pathology should be reexamined to ensure that the original tumor was diagnosed correctly. Electron microscopy can be useful in differentiating sarcomatous-appearing epithelial lesions from true soft-tissue sarcomas, and thus can be helpful in guiding therapy. PMID- 12092285 TI - Surgery-induced thyroiditis: fact or fiction? AB - Neck surgery carries a risk that the patient will develop postoperative thyroiditis as a result of the surgical procedure. Surgery-induced thyroiditis can manifest in a mild form as serum hyperthyroxinemia or in a more severe form as clinical hyperthyroidism. We describe a case of surgery-induced thyroiditis and review the very limited literature on this subject. PMID- 12092286 TI - Facial reanimation. PMID- 12092287 TI - Witnessing history in the making. PMID- 12092288 TI - Acceptance of death and dying. PMID- 12092290 TI - Make a difference with mentoring. PMID- 12092289 TI - Correct dosage? PMID- 12092291 TI - Assessing tissue oxygenation. AB - Although physical examination remains an important part of the nursing assessment, assessment of tissue oxygenation provides additional vital information. Nurses who are caring for patients with compromised functioning must have a basic understanding of the physiological relationships between oxygen saturation, cardiac output, and acid-base balance and must be able to plan nursing care based on this knowledge. PMID- 12092292 TI - Telemetry to home: successful discharge of patients with ventricular assist devices. PMID- 12092293 TI - Prone positioning of trauma patients with acute respiratory distress syndrome and open abdominal incisions. PMID- 12092294 TI - Reteplase: nursing implications for catheter-directed thrombolytic therapy for peripheral vascular occlusions. AB - Several studies since 1998 have shown the efficacy of catheter-directed thrombolytic therapy with reteplase. Reteplase is a plasminogen activator that penetrates the thrombus and causes lysis. This catheter-directed approach has been used to treat both arterial and venous occlusions, with a success rate of 72% to 88%. The most serious complication associated with thrombolytic therapy is intracranial hemorrhage. Patients should be admitted to the intensive care unit for monitoring of neurological status, vital signs, laboratory values (hematocrit, hemoglobin level, activated partial thromboplastin time, and fibrinogen concentration), and bleeding or oozing at puncture sites. Staff nurses in the intensive care unit must be aware of this important thrombolytic therapy, its indications, and its implications for nursing interventions. PMID- 12092295 TI - Capping arterial sheaths in patients undergoing percutaneous coronary intervention: evidence-based practice. PMID- 12092296 TI - Applying the synergy model to nursing education. PMID- 12092297 TI - How do I know if I have been sued as a nurse and what do I do? PMID- 12092298 TI - Patients who experience the common but uncomfortable sensation of "needing to void". PMID- 12092299 TI - Smaller tubes reduce the volume of blood drawn. PMID- 12092300 TI - [Persistent tachypnea in infants: possibly due to interstitial pneumonitis]. AB - In three male infants aged 3, 4.5 and 11 months with tachypnea and feeding problems, the initial supplementary examination revealed no possible cause. The tissue obtained by open lung biopsy showed interstitial pneumonia/pneumonitis. The two youngest patients were treated with hydrochloroquine and prednisone; the youngest died at the age of 18 months. Both the infant who was maintained on hydrochloroquine and the one who did not receive treatment showed a normal respiratory frequency and growth during the following years. Tachypnea is less easily recognised in infants than in older patients. It may be the only early symptom of interstitial pneumonitis. A normal chest X-ray cannot exclude an incipient interstitial pneumonitis as the cause of the tachypnea. A high resolution CT-scan of the lung parenchyma is necessary in order to detect the disease at an early stage. Histological examination of an open lung biopsy specimen is essential for the specific diagnosis, therapy and prognosis. PMID- 12092301 TI - [Oral fluoropyrimidines registered for the treatment of metastatic colorectal carcinoma: a possible gain]. AB - Up until now the standard treatment for metastasized colorectal carcinoma has been fluorouracil (5-FU) in combination with folonic acid in low doses administered intravenously, even after the recent registration of a number of new intravenously administered cytostatics, such as irinotecan and oxaliplatin. Meanwhile there are oral alternatives for 5-FU: capecitabine and the combination of tegafur and uracil with folonic acid. In four randomised studies it was shown that these drugs were globally just as effective as the combination of 5-FU with folonic acid (in accordance with the 'Mayo Clinic' scheme). There was no survival advantage for the oral drugs compared to 5-FU with folonic acid. Compared to 5-FU and folonic acid the use of capecitabine or tegafur-uracil-folonic acid was associated with less toxic effects; however, there were differences in the side effects profile between the oral drugs and 5-FU (more hand-foot syndrome for capecitabine and less (symptomatic) leucopenia for tegafur-uracil-folonic acid). An examination of the serious side effects (grade 3 and 4) revealed that the total incidence was generally comparable. These data, together with the ease of oral administration, form the basis for the registration of capecitabine and tegafur-uracil-folonic acid. The definitive place of these drugs in the treatment of metastasized colorectal carcinoma is not yet clear. PMID- 12092302 TI - [Clinical thinking and decision making in practice. A young woman with fever, arthralgia and exanthema]. AB - A 19-year-old woman was admitted because of high fever, rash, arthralgia and sore throat. On physical examination a diffuse skin rash was observed, leaving a facial mask unaffected. C-reactive protein and erythrocyte sedimentation rate were raised (114 mg/l and 26 mm in the first hour, respectively); white blood cell count was normal (6.2 x 10(9)/l) with an increased count of immature forms. An infective, metabolic or haematological cause was excluded. Serum ferritin turned out to be extremely elevated (4318 micrograms/l), so adult-onset Still's disease was diagnosed. The patient fulfilled the criteria of Cush et al. for adult-onset Still's disease. She was first treated with non-steroidal anti inflammatory drugs (NSAIDs) and, at a later stage in the disease, with corticosteroids. All symptoms disappeared and blood test results normalised. PMID- 12092304 TI - [Many psychiatric disorders in Afghan refugees with residential status in Drenthe, especially depressive disorder and post-traumatic stress disorder]. AB - OBJECTIVE: To assess the prevalence of psychiatric disorders and help-seeking behaviour of Afghan refugees with a residence status in the province of Drenthe, the Netherlands. DESIGN: Cross-sectional prevalence study. METHOD: By means of a modified multiflex snowball sampling with 3 primary informants, 55 Afghan households in 16 towns and villages in Drenthe were selected. After informed consent had been obtained, 51 adults completed a sociodemographic questionnaire and a culturally validated shortened 'Composite international diagnostic interview' (CIDI), DSM-IV version. Help-seeking behaviour was recorded. RESULTS: Half of the sample had resided in the Netherlands for at least 3 years. There was a high level of education, an unemployment level of 88% and moderate to poor language skills in 92% of the respondents. The prevalence of psychiatric disorders was 65% (95% CI: 52-78); depressive disorder occurred in 57% and posttraumatic stress disorder in 35%. Psychopathological findings were related to poor language skills, a lower level of education and unemployment. A total of 56% received treatment from a general practitioner and 4% were under psychiatric treatment. CONCLUSIONS: The prevalence of psychiatric disorders among adult Afghan refugees in Drenthe with a residence status was considerably higher than among the general population in the Netherlands. PMID- 12092303 TI - [Diagnostic image (92). A man with fever during chemotherapy. Invasive pulmonary mycosis]. AB - A 64-year-old male was treated for acute myeloid leukemia with idarubicin and cytarabin. He developed pulmonary mycosis (radiologically consistent with aspergillosis), which responded to intravenous amphotericin B. PMID- 12092305 TI - [Rising incidence of invasive (T4) head-neck carcinoma at the Utrecht University Medical Center, 1980-1998]. AB - OBJECTIVE: To determine whether there is an increase in the incidence of diagnosed head and neck tumours at an advanced stage (T4). DESIGN: Secondary data analysis. METHOD: Data of patients registered in 1980-1998 at the Utrecht University Medical Centre, the Netherlands, were analysed for the relative prevalence of T4 tumours in the oral cavity, pharynx and larynx on the basis of sex, age and tobacco and alcohol usage. The results were compared with figures from the Dutch national cancer registry (1989-1997). RESULTS: Data were collected for 2993 patents (2256 men and 737 women; average age 63 years (SD 12)). Of these 35 had a carcinoma situated on the lips, 1091 in the oral cavity, 1321 in the larynx and 546 in the pharynx. There was an increase in T4 tumours of 12.2% (43/353) of all tumours in 1980-1984 to 27.2% (252/1033) in 1995-1998. Linear regression analysis estimated an increase of 0.9% every year. Selected referral to the Utrecht University Medical Centre was unlikely because the increased incidence of T4 tumours could be recognised in the national cancer registry. There was no change in the sex distribution over the study period. The increase in the 40 to 50-year-old age group was greater than that of the patients aged 80 years and over. T4 tumours were more prevalent than T1 tumours in patients who had smoked 20 or more cigarettes per day or drank more than 4 units of alcohol per day. CONCLUSION: The percentage of cancer patients presenting with an advanced stage (T4) of head-neck tumours increased during the study period. PMID- 12092306 TI - [Parinaud's syndrome as a sign of acute obstructive hydrocephalus: recovery after acute ventriculostomy]. AB - Paresis or palsy of upward vertical gaze, pupillary light-near dissociation and nystagmus retractorius with convergence, were present in a boy aged 16 years and a woman aged 30 years with an obstructive hydrocephalus due to an aqueductal stenosis as a consequence of a bacterial meningitis and in a woman aged 26 years and a man aged 47 years with an outlet obstruction of the fourth ventricle after a posterior fossa operation for a tumour in the fourth ventricle. All of the patients were suspected of having a drain dysfunction. They all underwent a third ventriculocisternostomy after which their symptoms (partially) resolved. The presenting symptoms stated are the classical triad of Parinaud's syndrome. In addition to these there are less frequent symptoms such as bilateral eyelid retraction (Collier's sign) and convergence spasms. The syndrome is rare but has a significant mortality risk and a high morbidity rate if an obstructive hydrocephalus is not diagnosed and treated. An MRI scan of the cerebrum to detect obstructive hydrocephalus with dilation of the aqueduct is the diagnostic of choice. For an obstructive hydrocephalus with dilation of the aqueduct a third ventriculocisternostomy is the treatment of choice. PMID- 12092307 TI - [Lamivudine for the prevention of chronic hepatitis B exacerbations during chemotherapy for non-Hodgkin's lymphoma]. AB - Two male patients, aged 54 years and 17 years respectively, were treated with chemotherapy for non-Hodgkin lymphoma. Both patients were chronic hepatitis-B virus (HBV) carriers prior to the chemotherapy. One patients died as a result of the virus exacerbating during chemotherapy; the other patient received the antiviral drug lamivudine prior to the chemotherapy and finished the cures without any problems. Exacerbations of HBV replication followed by an increase in serum transaminase activity levels ('flares') occur naturally during the course of the viral infection. However, there is an elevated risk when a patient receives high doses of corticosteroids for a short period, as is the case in chemotherapy for non-Hodgkin's lymphoma. Lamivudine is registered for the treatment of chronic hepatitis B and can be used as a prophylactic prior to chemotherapy or to treat an exacerbation of the hepatitis. It is advisable to systematically test all patients with lymphoma for the presence of the HBsAg. If this is positive, prophylactic administration of lamivudine 100 mg once daily is strongly recommended if chemotherapy is indicated. PMID- 12092308 TI - [One hundred years of the Association of Surgeons in the Netherlands. V. Traumatology]. AB - Traumatology in the Netherlands gained considerable momentum during the last years of the twentieth century. This breakthrough was due to the creation of ten trauma care centres, each with its own 'mobile medical team'. Additionally, four of these centres dispose of a helicopter for rapid transport of this team. Furthermore, within the field of medical education and training, new courses on advanced trauma and life support (ATLS) and advanced pediatric life support (APLS), for example, not only offer more practical knowledge, but also provide a 'common language' for all those involved in the medical and organisational aspects of trauma care. Surgical treatment of trauma injuries is becoming increasingly oriented towards minimally invasive techniques. Assessment of the quality of care provided requires a broad and reliable registration system for injuries. PMID- 12092309 TI - [Medical education in Egypt]. AB - Modern medical training in Egypt was started by Antoine Clot Bey in 1837 and became part of the university programme in 1919. At present, it comprises six years of university education, followed by one year of internships and one year of compulsory employment with a state-owned hospital. There are now 13 medical faculties in Egypt, using three different curricula: traditional, Islamic and innovative. Their implementation is hampered by the large number of students (15,500 men and 7500 women), the low salaries and motivation of the instructors, the teaching in English rather than Arabic and the lack of recent study materials. It is therefore rather difficult to compare the effectiveness of the Egyptian system with that in the Netherlands. Due partly to the differences in language and culture, Dutch authorities are reluctant to recognise Egyptian medical diplomas. PMID- 12092310 TI - [The automatic exterior defibrillator in the Netherlands: advice of the Dutch Health Council]. PMID- 12092311 TI - [Treatment of patients with differentiated thyroid carcinoma]. PMID- 12092312 TI - [Gluteal abscess complicated by sepsis presenting as Crohn's disease]. PMID- 12092313 TI - [Gluteal abscess complicated by sepsis presenting as Crohn's disease]. PMID- 12092314 TI - [Bilateral pneumothorax in a young woman after acupuncture]. PMID- 12092315 TI - [Physical diagnosis--rectal palpation]. PMID- 12092316 TI - Pathophysiology of upper airway obstruction during sleep. AB - The obstructive sleep apnoea syndrome (OSAS) is characterised by repetitive upper airway obstructions leading to snoring, hypopnoeas and apnoeas. Sleep fragmentation and arterial oxygen desaturations are the first consequences. Since the discovery of the OSAS, an impressive series of studies were done on upper airway physiology, and more specifically, on the pharyngeal airway. The objective of this article is to review the pathophysiological basis of upper airway obstruction during sleep. PMID- 12092317 TI - Clinical presentation and definitions of sleep-related breathing disorders. AB - The sleep disordered breathing (SDB) patients need to be monitored for sleep and breathing during sleep. However there is no consensus about the definitions of the most common features observed during the night. Although apnea is usually defined as a cessation of airflow lasting more than 10 seconds, the definitions of hypopnea differ in the literature with respect to the amount of reduction of airflow and to the drop in oxygen desaturation. Variations in the technique of monitoring airflow and ventilary effort can cause variation in the detection of SDB events. We discuss in this paper the standard definitions of respiratory events as observed during the night and the elements of diagnosis obtained with the polysomnographic examination for the SDB patients. PMID- 12092318 TI - Acoustic analysis of snoring: review of literature. AB - Acoustic analysis of snoring could make patient assessment more accurate, treatment planning more efficient and could provide objective documentation reliable for comparison. However, spectral characteristics are irregular and difficult to analyse. The authors give an overview of the available literature and the various applied methods. These methods differ in purpose and are subject to restrictions. Until now, acoustic analysis of snoring remains controversial. For linking the results of the acoustic analysis with anatomical levels or degree of obstruction, further study is needed to develop a reliable parameter. PMID- 12092319 TI - Nasal patency and sleep-related breathing disorders. AB - This article reviews the current insights in the contribution of nasal physiology, resistance and pathophysiology in relation to sleep-related breathing disorders. Different possible pathways have been followed to prove a clear role for the nose in this matter; they are reviewed and discussed. From all this it should be concluded that today there is only evidence for abnormal nasal resistance to be a possible cofactor in the disease. A short reminder about the currently available diagnostic tools for assessing nasal patency and resistance is given next. Finally the relation between the nose and treatment of sleep related breathing disorders with nocturnal continuous positive airway pressure is discussed. PMID- 12092320 TI - Upper airway collapsibility: measurement techniques and therapeutic implications. AB - Several techniques are currently available that aim to characterize upper airway function/mechanics during wakefulness or sleep. Based on the concept of a Starling resistor, we propose a standardized protocol to measure the critical pressure (Pcrit) (an indicator of upper airway collapsibility) during sleep. The effect of therapeutic interventions such as weight loss, positional changes or uvulopalatopharyngoplasty on Pcrit is illustrated by data from the literature. We propose that measurement of Pcrit become implemented in the diagnostic work-up of selected patients with sleep-related breathing disorder to help making a correct therapeutic decision. PMID- 12092322 TI - Upper airway imaging in patients with sleep-related breathing disorders. AB - This paper focuses primarily on upper airway anatomy and its variations during breathing and highlights the advantages/disadvantages of the different imaging techniques. The major indications of upper airway imaging are also discussed. PMID- 12092323 TI - Conservative and non surgical treatments for the sleep disordered breathing adult. AB - Obstructive sleep apnoea (OSA) syndrome is a chronic disease characterised by repetitive obstructive apnoeas and hypopnoeas during sleep leading to sleep fragmentation. The treatment of OSA consists of general and specific measures. The aim of this article is to review the different non surgical approaches to treat OSA in adults. The different therapeutical options can be used in combination, which underlines that the OSA patient needs a careful and multidisciplinary management. PMID- 12092321 TI - Standard ENT clinical evaluation of the sleep-disordered breathing patient; a consensus report. AB - Sleep disordered breathing (SDB) patients usually undergo an ENT clinical examination before any therapeutic decision. This clinical examination would be predictive about the occurrence of Obstructive Sleep Apnea Syndrome, cost effective, reproducible and would determine the sites of obstruction in the upper airways. To achieve this, ENT specialists from Belgium, representatives of academic hospitals in the country, have tried to establish an updated work-up in the clinical evaluation of the SDB patients. History, risk factors evaluation, excessive daytime sleepiness, static and dynamic evaluation of the upper airways (velopharynx aspect, tonsils grading, Mallampati score, Muller's maneuver, fiberoptic evaluation) have been standardized in a consensus report easily accessible to the vast majority of ENT specialists. This consensus must be understood as a clinical work-up to perform before the monitoring of breathing during sleep. PMID- 12092324 TI - Effect of nasal valve surgery by open-septorhinoplasty and lateral cartilage grafts (spreader grafts) on snoring among a population of single snorers. Preliminary report. AB - Snoring is traditionally considered more of a social nuisance than disease. The observation that nasal obstruction may modify breathing during sleep and daytime behavior and our clinical observations suggesting that some single snorer patients who had undergone surgery of the nasal valve for nasal obstruction complaints had presented a decrease in their snoring, led us to make a retrospective study on twenty nine patients. Eight of them were simple snorers. They underwent a surgery of the nasal valve under general anesthesia through an open septorhinoplasty during which lateral graft cartilages (modified spreader grafts) were set in place between septolateral cartilage and the anterior and superior edge of the nasal septum. Active anterior rhinomanometry (AAR), posterior active rhinomanometry (PAR) and a self-assessment of the nasal patency by the patient himself on a visual analog scale (VAS) were compared before and one year after surgery. Snoring was assessed by both bed partners according to a four step scale (none, slight, moderate and severe). No significant differences were found between AAR or between PAR. But significant differences were measured with VAS (for the left (p < 0.0001) and the right sides (p < 0.001). All patients but one demonstrated substantially decreased snoring. As there is no clear cut evidence of changes in nasal resistances measured by rhinomanometry, we suggest that the observed decrease in snoring may be induced by changes in the direction of the nasal airflow against the nasal mucosa which may produce by a reflex between mecanoreceptors of the nasal mucosa and muscles of the pharynx, an increased tonus of the muscles of the pharynx. PMID- 12092325 TI - UPPP for snoring: long-term results and patient satisfaction. AB - We retrospectively survey 57 patients who underwent uvulopalatopharyngoplasty (UPPP) because of habitual snoring over a five-year period. A total of 100 patients were sent questionnaires concerning persistent snoring, excessive daytime sleepiness (EDS), body mass index (BMI) and postoperative complications. After 5 years the success rate was 53%. The mean snoring score was 16.7 preoperatively and decreased to 10.6 postoperatively. There was a relationship between the body mass index (BMI) preoperatively and the efficiency of the surgery. The mean EDS score was 11.1 before and 9.4 after surgery. UPPP in patients complaining of snoring is quite successful but the results decline significantly with time and patients should be warned of the possibility of snoring remaining or returning. PMID- 12092326 TI - Side effects and complications of velopharyngeal surgery. AB - Although velopharyngeal surgery is generally safe, complications are not uncommon. This article reviews the major complications occurring in the postoperative period and the severe late complications such as velopharyngeal incompetence and nasopharyngeal stenosis. Most of these complications can be prevented by a careful operative technique and preoperative evaluation. We also emphasise the frequency of minor modifications of voice, swallowing, taste and velopharyngeal function. PMID- 12092327 TI - Laser-assisted surgery addressing snoring long-term outcome comparing CO2 laser vs. CO2 laser combined with diode laser. AB - Our study encompasses 61 patients (49 men--80.3%; 12 women--19.7%) treated from September 1998 through August 2001. Mean follow-up covers 25 months (range: 7-43 months). Our CO2-LAUP technique involves vaporizing the palatine mucosa along a rectangular surface from the palatal dimple to the base of the uvula; trimming the palatine arches under the velum; and resecting the uvula. For 22 patients, we employed intravelar diode laser coagulation in the expectation of reducing the postoperative pain whilst achieving the same therapeutic effectiveness. There is no pain difference between the CO2-LAUP technique versus that combining intravelar diode laser coagulation with CO2 laser uvula resection and trimming of the palatine arches. Mean maximal pain reaches 6.93 +/- 3.55 with CO2 laser and 6.95 +/- 3.64 with CO2 laser plus diode laser. Similarly, both techniques involve the same mean algesic period of 22 days with the day of maximal pain at 1 week after surgery. Associating base of tongue vaporization significantly increases the algesic period (p = 0.042). No long-term complications were observed in relation to intravelar coagulation or LAUP, whether combined or not with base of tongue vaporization. In terms of patients satisfaction, no significant difference exists between the various surgical techniques of the velum alone. The satisfaction rate reaches 5.26 +/- 3.92 with CO2 laser and 5.82 +/- 2.67 with the CO2 laser plus diode laser. Satisfaction is statistically identical when base of tongue vaporization is included in the procedure. PMID- 12092328 TI - Radiofrequency volumetric reduction of the palate in patients with sleep related breathing disorders. PMID- 12092329 TI - Parapharyngeal causes of sleep apnea syndrome: two case reports and review of the literature. AB - Most patients with obstructive sleep apnea experience increased pharyngeal collapsibility which predisposes them to upper airway occlusion during sleep. Some patients with or without a higher-than-normal collapsibility may present other causes of sleep apnea syndrome. This article will focus on the parapharyngeal tumors leading to obstructive sleep apnea (OSAS). Two clinical cases will illustrate our review of the literature. The results of surgery will also be discussed. PMID- 12092330 TI - Surgical treatment of the sleep-disordered breathing patient; a consensus report. AB - Sleep disordered breathing patients may undergo surgical treatment after history, clinical examination and polysomnographic study if they demonstrate upper airway obstruction. This article focus on the surgical treatment designed for these patients. Sino-nasal surgery, rhinopharyngeal procedure, velopharyngeal procedures (Uvulopalato-pharyngoplasty, Laser assisted uvulopalatoplasty, Radiofrequency tissue volume reduction) as well as base of the tongue procedures were discussed among a panel of Belgian ENT specialists offering their experience in this field. Algorithm on corrective surgery as well as guidelines for postoperative management are proposed in the management of sleep disordered breathing patients. PMID- 12092331 TI - Apnea and snoring: state of the art and future directions. AB - This review is intended to discuss the newer aspects of sleep apnea, and particularly where the new advances are likely to take place in the current millennium. The topics reviewed include epidemiology and clinical features of sleep apnea and snoring, adverse health consequences of sleep-disordered breathing, upper airway resistance syndrome, and treatment. Most of the work reviewed in these areas has been published since the year 2000, although whenever necessary older literature on the topic under discussion is summarized. Based on the accumulated information to date, new advances in the sleep-disordered breathing are likely to occur in the genetics of sleep apnea, identification of cellular mechanisms involved in control of vascular resistance, and pharmacological treatment of sleep apnea directed toward specific target such as pharyngeal muscles. PMID- 12092332 TI - Velopharyngeal anatomy in snorers and patients with obstructive sleep apnea. AB - Velopharyngeal structures play an important role in the pathogenesis of snoring and obstructive sleep apnea (OSA). Hence they form a tempting target for surgical interventions in the treatment of these sleep-related breathing disorders. The assessment of the patient with snoring should therefore include a thorough evaluation of the velopharynx. The clinical evaluation of the velopharynx is discussed in normals and patients who snore (with or without OSA), as well as the features obtained using cephalometry and CT and MR imaging. PMID- 12092333 TI - [Illness caused by the profession]. PMID- 12092334 TI - [Risk and risk perception]. AB - After having delineated the concept of risk and having described the type of risks that are relevant to dental practice, an overview of empirical and theoretical work on risk perception is given. The perception of risks is determined both by the actual magnitude and by perceptual bias, originating from either general perceptual phenomena such as (cognitive) availability or the ego relevance of risks: persons in general underestimate risks for themselves as compared to the risks for others (unrealistic optimism). PMID- 12092335 TI - [Dentist and disability: a matter of occupational disease?]. AB - The incidence of and background to disability claims by dentists relating to 1999 are presented on the basis of data from Movir insurance company. Possible cases of occupational related disease are indicated. Claims resulting in more than 2 weeks sick leave have been received from almost 7% of the insured dentists (excluding pregnancy related sick leave). In nearly 30% of the cases, sick leave lasted for more than 1 year (particularly due to mental and musculoskeletal disorders). Compared to the figures for 1992 long-term health related absenteeism (longer than 365 days) has increased by as much as 155%. It is estimated that at least 15% of the claims are occupationally related. Based on the 1999 data a dentist would have approximately a chance of 50% for partial or complete premature retirement due to ill health. PMID- 12092336 TI - [Career expectations and the type of dentist in the light of burnout]. AB - The aim of this study was to measure if dentists with unfulfilled career expectations have higher burnout scores than dentists who succeeded in realising their expectations. Furthermore, the relation between type of dentist and burnout was investigated. It appeared that dentists who experienced reality being worse than expected, showed higher burnout levels, as did dentists putting more interest in life outside practice. It is recommended that attention for realistic career expectations is a regularly recurring topic in dental education. Also, introspection upon the type of dentist one prefers to be is something a professional should not neglect. PMID- 12092337 TI - [Complaints by complainers?]. AB - For Dutch physicians being sued is one of their worst nightmares in professional life. Dutch dentists as a group do not seem to bother. This article shows that the light-heartedness of the dental profession is incorrect. A Dutch dentist has the same reason for being worried about the chance of facing a (disciplinary) council, about the procedure and about (disciplinary) measures as Dutch physicians have. For dentists who actually had to face a disciplinary council, their experience was very frightening and affected their professional satisfaction. Knowledge of public health legislation and a basic knowledge of legal procedures can be useful weapons in the battle against burnout. PMID- 12092338 TI - [Aggression in the dental office]. AB - In this study on aggression in the dental practice, 8 dentists and 1 dental nurse who had experienced aggression in their dental practice, were interviewed and filled out a questionnaire. They had responded to a call in 2 Dutch dental journals or were contacted through personal communication. The interviews show that aggression in particular can be expected in patients having not paid their expense account, non compliant patients, or in patients who often come too late or even do not show up at their appointments. Aggression also can appear when patients with toothache are told they cannot be helped immediately. Aggression seems predominantly connected to people under influence of alcohol or drugs, psychiatric patients and people having contacts in criminal circuits. Although the incidence seems relatively low, the occurrence of aggression always has a great impact on the dental team. Therefore it seems useful to study how dentists and their auxiliaries could act to prevent aggression. PMID- 12092339 TI - [Profession-related disorders of the locomotor apparatus. A multi-causal problem?]. AB - This article provides information on the prevention of disorders among dentists and dental hygienists, for instance neck, shoulders, upper extremity, wrist and hand disorders. Many daily (strained) operating postures are described, and research data from the Sonde prevention project is mentioned. This data is compared with the newly implemented NEN-ISO-standard 11226. The standard lists the limits of static working postures. It becomes clear from this data that many dentists exceed these limits daily and work in overstrained postures. The article will further go into the complicated interrelationship of and connection between various interactive factors which intentionally and often unintentionally lead to static use of muscles. It may be a matter of cumulative effects that are multi causally related, with the possible consequences. A further awakening within the dental profession, conditioning of young students into adopting a correct operating posture and purchasing ergonomically sound equipment may contribute to a healthier situation. PMID- 12092340 TI - [Skin reactions to dental materials]. AB - In dentistry, contact with a wide range of both irritants and contact allergens pose an important occupational hazard. It is important to understand the resulting clinical skin symptoms and their causes to prevent further exposition and to avoid serious problems that may lead to an incompatibility with the dentist profession. PMID- 12092341 TI - [Mortality of dentists]. PMID- 12092342 TI - [Food additives and the quality of food]. PMID- 12092343 TI - [What kinds of food products are functional?]. PMID- 12092344 TI - [Genetically modified and other novel food products]. PMID- 12092345 TI - [Are vegetarian diets safe?]. PMID- 12092346 TI - [Microscopical colitides as a cause for chronic diarrhea]. PMID- 12092347 TI - [Ethylene glycole poisonings in Nothern Savo 1984-98]. PMID- 12092348 TI - [And the clear mucus secretion continued and continued....]. PMID- 12092349 TI - [A hydroxyurea-induced leg ulceration]. PMID- 12092350 TI - [Treatment of hallux valgus]. PMID- 12092351 TI - [Irritable bowel or colitis?]. PMID- 12092352 TI - [A lump in a child's neck]. PMID- 12092353 TI - [Art and science and the medical profession]. PMID- 12092354 TI - [Good food]. PMID- 12092355 TI - [Sense impression of the quality of food products and changing liking for food]. PMID- 12092356 TI - [Homeostasis of gastrointestinal system and the bioavailability of nutritive substances]. PMID- 12092358 TI - [Natural or supplemented food products?]. PMID- 12092357 TI - [Can our food be made healthier by special supplements?]. PMID- 12092359 TI - [Microbiological dangers of food]. PMID- 12092360 TI - [Organic environmental toxins in food products and their way into the human]. PMID- 12092361 TI - [Can allergy be prevented by the means of nutrition?]. PMID- 12092362 TI - [Pregnant and lactating women--a new nutritional risk group]. PMID- 12092363 TI - [Ribosome proved to be a RNA enzyme]. PMID- 12092364 TI - [Whose bread you eat, his songs you will sing?]. PMID- 12092365 TI - [Nutrition during pregnancy--do recommendations and practice meet?]. PMID- 12092367 TI - [Nucleoside analogues--new drugs for the treatment of lymphatic cancers]. PMID- 12092368 TI - [Progress file in a specialist or completing further education of pediatrics]. PMID- 12092366 TI - [Bronchitis and emphysema as occupational diseases]. PMID- 12092369 TI - [Persistent insomnia in a depressed athlete]. PMID- 12092370 TI - [Are vitamins dangerous?]. PMID- 12092372 TI - [How, when and why do the drugs go out of date]. PMID- 12092371 TI - [Uncontrollable hypercalcemia in a dialysis patient--parathyromatosis or parathyroid carcinoma?]. PMID- 12092373 TI - [Sentinel lymph nodes in breast cancer]. PMID- 12092374 TI - [Fever and erythema nodosum]. PMID- 12092375 TI - [In "the doctor in his place" article series knowledge, understanding and energy to continue in the profession meet]. PMID- 12092376 TI - [Make the pagination visible]. PMID- 12092377 TI - [Coeliac disease and lymphoma]. PMID- 12092378 TI - [Mad cows in media and in medicine]. PMID- 12092379 TI - [Oral fungal infections]. PMID- 12092380 TI - [Angelman syndrome--from the phenotype of a developmental disorder to the genes]. PMID- 12092381 TI - [Intensity modulated radiotherapy--new technique that will probably improve treatment results]. PMID- 12092382 TI - [Depression after cerebrovascular disorders]. PMID- 12092383 TI - [Subcutaneous electrocardiographic loop recorder in the investigation of syncope]. PMID- 12092384 TI - [Forced to do--forced to think]. PMID- 12092386 TI - [Diagnostics and treatment of botulism]. PMID- 12092385 TI - [A rare spinal damage in a child]. PMID- 12092387 TI - [The one who does, does also mistakes]. PMID- 12092388 TI - [Treatment decision is the physician's most difficult decision]. PMID- 12092389 TI - [Heavy work, heavy amusements]. PMID- 12092390 TI - [Fungal infections of the skin, hair and nails: sample collecting, diagnostics and reply praxis]. PMID- 12092391 TI - [Treatment of acute respiratory insufficiency without intubation]. PMID- 12092392 TI - [Are dioxins a health problem in Finland?]. PMID- 12092394 TI - [Paragangliomas of neck and temporal bone]. PMID- 12092393 TI - [Tumor necrosis factor alpha and the heart--can heart diseases be treated by neutralizing cytokines?]. PMID- 12092395 TI - [Antenatal screening methods for chromosome 21 trisomy]. PMID- 12092397 TI - [Acalculous cholecystitis in a previously healthy man]. PMID- 12092396 TI - [Facial eczema in a jigger]. PMID- 12092398 TI - [Examining a patient with dyspnea on duty]. PMID- 12092399 TI - [What was the problem with hallux valgus?]. PMID- 12092400 TI - [Helene Schjerfbeck: Mother and child 1866]. PMID- 12092401 TI - [Why do allergic diseases increase?]. PMID- 12092402 TI - [Asthma from childhood to adolescence]. PMID- 12092403 TI - [How is an allergic immunological response formed?]. PMID- 12092404 TI - [Diagnostics of milk allergy]. PMID- 12092405 TI - [Asthma in early childhood]. PMID- 12092407 TI - [Atopic eczema--an allergy or not?]. PMID- 12092406 TI - [A sniffing child]. PMID- 12092408 TI - [Imaging diagnosis of dementia. Nuclear medicine]. PMID- 12092409 TI - [Imaging diagnosis of dementia. Brain CT]. PMID- 12092410 TI - [Imaging diagnosis of dementia. MRI]. PMID- 12092411 TI - [Simultaneous determination of five antioxidants in food by HPLC with fluorescence detection]. AB - An HPLC method with fluorescence detection was developed for the determination of propyl gallate, nordihydroguaiaretic acid, butylated hydroxyanisole (2- and 3 tert-butyl-4-hydroxyanisole), tert-butylhydroquinone and octyl gallate in edible oils and foods. The antioxidants in edible oil were isolated directly with acetonitrile saturated with n-hexane. The antioxidants in food were extracted with ethyl acetate and the extract was concentrated under vacuum. They were isolated from the residue with acetonitrile saturated with n-hexane. The acetonitrile layer was centrifuged at 5,000 rpm for 10 min. The HPLC separation was performed on a Symmetry C18 column (3.5 microns, 4.6 mm i.d. x 150 mm) using a mixture of 5% acetic acid-acetonitrile-methanol (4:3:3, v/v/v) as the mobile phase and monitored by using a fluorescence detector with time programming. Sample peaks were identified by comparison of the fluorescence spectra with those of antioxidant standards. Average recoveries of fortified antioxidants at 100 micrograms/g were 72.1-99.6%. Coefficients of variation were 0.7-7.2%. PMID- 12092412 TI - [Analysis of lac color in diets and feces of rats for toxicity studies]. AB - An analytical method was developed for lac color in diets fed to rats and in the feces, and the contents of lac color were determined. After lac color was extracted with 0.05% sodium carbonate and 50% ethanol containing 0.02% sodium lauryl sulfate from the diets and feces, the extracted color solutions were analyzed by HPLC. The recoveries of lac color from diets spiked at 1.25, 5.00% and that from feces spiked at 5.00% were 85.6, 93.4% and 69.5%, respectively. Contents of lac color in diets prepared to contain 1.25 and 5.00% were 1.1 and 5.2%, and dose levels were confirmed by these results. Contents of lac color in feces of male and female rats given lac color were 127.8 mg/g and 138.6 mg/g, respectively. By comparing the HPLC chromatograms of laccaic acids in the diet with those in feces of rats, laccaic acid A, B, C and E were detected in both, and their content ratios were approximately determined. PMID- 12092413 TI - [Survey of the daily intake of nitrate and nitrite in school children by the duplicate portion method]. AB - The daily intake of nitrate and nitrite in school children (n = 100) in Hiroshima Prefecture was estimated directly by the duplicate portion method. The daily intake of nitrate was 68.42 +/- 77.49 mg. The daily intake of nitrate/kg body weight was 2.06 +/- 2.24 mg, which is about 56% of the acceptable daily intake (ADI). The daily intake of nitrite was 0.953 +/- 0.869 mg. The daily intake of nitrite/kg body weight was 0.027 +/- 0.021 mg, which is about 45% of the ADI. The daily intake of nitrite/kg body weight was significantly different between the obese group and the non-obese group in boys (p < 0.05, Mann-Whitney U-test). The rates of children whose daily intakes of nitrate and nitrite were above the ADI were 16% and 7%, respectively. PMID- 12092414 TI - [Oxytetracycline and oxolinic acid residues in kuruma prawn (Penaeus japonicus) and the effect of cooking procedures on the residues]. AB - Tissue distribution and residue depletion of oxytetracycline (OTC) and oxolinic acid (OA) were studied in the kuruma prawn (Penaeus japonicus). The prawn were kept in tanks with recirculated artificial seawater at a salinity of 22-23@1000. The water temperature was maintained at 25 degrees C. The average body weight was 22.9 +/- 4.9 g for OTC and 22.5 +/- 3.6 g for OA. The drug was mixed with the diet and orally administered through a catheter to the prawn. The doses of OTC and OA, respectively, were 50 mg/kg body weight. At each sample time, four prawns were sacrificed and tissues were sampled. OTC and OA levels were determined by high-performance liquid chromatography. At the highest levels, the concentrations of OTC were in the other: shell (13.57 micrograms/g) > hemolymph (12.20 micrograms/mL) > muscle (8.30 micrograms/g). For OA, the order was: shell (20.74 micrograms/g) > hemolymph (7.06 micrograms/mL) > muscle (2.05 micrograms/g). The elimination half-lives of hemolymph and muscle were 44.7 and 46.8 hours for OTC and 55.0 and 107.9 hours for OA, respectively. Residual OTC could not be detected in hemolymph and muscle at 20 days after dosing. Residual OA disappeared from hemolymph and muscle at 25 days after dosing. A 25-day period for OTC and 30-day period for OA could be regarded as the proper withdrawal time established for kuruma prawn by the Pharmaceutical Law in Japan. However, the elimination half lives of shell for OTC and OA could not be calculated because both drug residues persisted in shell tissues, and the elimination phase was not completed during the experimental period. Residual OTC (14.10 +/- 2.26 micrograms/g, n = 6) and OA (0.32 +/- 0.06 microgram/g, n = 7) were detected in exuviae at 3 days and 4 days after dosing, respectively. Residual OTC was reduced to 50-70% in muscle by the usual methods of cooking (boiling, baking at 200 degrees C and frying at 180 degrees C), whereas reduction levels in shell were only 20-30%. Residual OA was reduced to 20-30% in muscle and shell by the cooking. These results confirm that the cooking procedures could only reduce but not completely eliminate these drug residues in prawn. PMID- 12092415 TI - Increased digestibility of two products in genetically modified food (CP4-EPSPS and Cry1Ab) after preheating. AB - We performed experiments on in vitro digestion of newly expressed proteins by SGF (simulated gastric fluid) and SIF (simulated intestinal fluid) to assess the allergenicity of food components derived from biotechnological modification. For newly expressed proteins, we chose CP4-EPSPS (5-enolpyruvylshikimate-3-phosphate synthase from Agrobacterium sp. strain CP4) and Cry1Ab derived from Bacillus thuringiensis subsp. kurstaki strain HD-1. The former is expressed in GM-soybeans and the latter is expressed in GM-corns. Firstly, we examined the digestibility of purified CP4-EPSPS and Cry1Ab by SGF. Both proteins were rapidly digested within 60 sec. After preheating, the digestibility by SGF was slightly increased. Secondly, CP4-EPSPS in GM-soybean extracts and Cry1Ab in GM-corn extracts were digested by SGF. The digestion time of both proteins by SGF was almost the same as that of the purified proteins. Thirdly, the digestibility of CP4-EPSPS and Cry1Ab by SIF was examined. The digestion time of these proteins was 240 min or more. However, digestibility of these proteins by SIF was dramatically increased by preheating, and the digestion time was less than 5 sec. Fourthly, CP4-EPSPS in GM-soybean extracts and Cry1Ab in GM-corn extracts were digested by SIF. Digestion time of both proteins by SIF was almost the same as that of the purified proteins. From these results, we concluded that the digestibility of both CP4-EPSPS and Cry1Ab by SGF and SIF was increased by preheating. Therefore, we suggest that the allergenicity of both proteins should be extremely low because of the easy digestibility of these proteins by SGF and also by SIF with preheating. PMID- 12092416 TI - [Detection of unexpected recombinant DNA in maize grain]. AB - PCR detection of genetically modified (GM) line-specific recombinant DNA was carried out on Bt 11, Event 176 and Non-GM maize grain harvested in 1999. Of 100 grains of Bt 11 line, Event 176 specific DNA was detected in 11 grains. Of 30 grains of Event 176 line, Mon 810 or Bt 11 specific DNA was detected in 5 grains. In addition, Bt 11 or Event 176 specific DNA was detected in 4 of 30 Non-GM grains. These data suggest that maize grains (not seed) often contain DNAs of different lines from what they were expected to contain. Furthermore, quantitative PCR was performed to estimate the genotype of the Event 176 grains described above. The results showed that the genotype of all the grains containing different recombinant DNA is heterozygous. Therefore, it was considered that the grains containing the unintended recombinant DNA were not accidental contaminants, but that airborne pollination had introduced the recombinant DNA into the grains. PMID- 12092417 TI - [Multiresidue analysis of nitrogen-containing and sulfur-containing pesticides in agricultural products using dual-column GC-NPD/FPD]. AB - We investigated simultaneous analytical methods for pesticide residues in large numbers of agricultural products samples. Extraction of each sample with acetonitrile was followed by a salting-out step using a graduated cylinder. The test solution was cleaned up with gel permeation chromatography (GPC), which separated the pesticide eluate into 2 fractions, and then with a tandem mini column. Analysis was done with a dual-column GC equipped with a dual NPD and FPD (S mode) detector. Use of the Siltek-deactivated liner, guard column, and Y connector, and Silcosteel-treated NPD jet was effective for preventing the breakdown of sulfur-containing pesticides. Recoveries of 87 nitrogen-containing and sulfur-containing pesticides from fortified spinach, tomato, apple, strawberry and brown rice, ranged from 71 to 127% with RSD values of 1-24%, except for recoveries of aldicarb, amitraz, ethiofencarb, imazalil, propamocarb and triflumizole. Detection limits of pesticides were very good (0.3-5 ppb (NPD) and 2-20 ppb (FPD)) for routine analysis of pesticide residues in foods. Surveillance of pesticides in agricultural products was carried out by using the present method. From 22 out of 33 samples, 21 pesticides (43 in total) were detected. The results indicated that the present method can be applied as an efficient and reliable means for monitoring pesticide residues in agricultural products. PMID- 12092418 TI - [Survival of Vibrio parahaemolyticus serovar O3:K6 strains under acidic conditions]. AB - The survival of Vibrio parahaemolyticus serovar O3:K6 strains and other serovars in the presence of acetic, citric and hydrochloric acids were studied. There were no differences in resistance to these acids between serovar O3:K6 and the other serovars. At pH 5.6, citric acid was more effective in reducing the number of viable cells of V. parahaemolyticus than acetic acid. However, at pH 4.5, acetic acid was more effective than citric acid. The number of viable cells decreased quickly in the presence of rice vinegar or wine vinegar at pH 4.0. PMID- 12092419 TI - [Simultaneous analysis of four diuretic drugs by HPLC and its application to health food supplements advertising weight reduction]. AB - A high-performance liquid chromatographic (HPLC) method for the simultaneous analysis of triamterene, trichlormethiazide, furosemide and spironolactone is presented for application in the examination of health food supplements advertising weight reduction and in the analysis of pharmaceuticals. The HPLC assay was performed under gradient conditions using a Wakosil ODS 5C18 column (5 microns, 150 x 4.6 mm i.d.). The mobile phase consisted of a gradient program with a mixture of water and acetonitrile containing 0.1% triethylamine adjusted with phosphoric acid to pH 3.0: from 0 to 6 min, 15% acetonitrile; from 6 to 20 min, linear gradient from 15 to 50% acetonitrile; and from 20 to 40 min, 50% acetonitrile. The column effluent was monitored from 0 to 20 min at 260 nm and from 20 to 40 min at 235 nm. The calibration curves of the four drugs showed good linearity and the correlation coefficients were better than 0.999 in all cases. The lower limits of detection were approximately 40 ng for each drug. Commercially available health food supplements and pharmaceuticals were analyzed after extraction with a mixture of methanol and acetic acid (99:1). The procedure described here is suitable for the screening of four diuretic drugs in adulterated supplements and for the quality control of pharmaceuticals with minimal sample preparation. PMID- 12092420 TI - Determination of ten sulfonyl urea herbicides in unpolished rice by solid-phase extraction cleanup and LC-diode array detection. AB - A liquid chromatographic (LC) method with diode array detection (DAD) was developed for screening of 10 sulfonyl urea herbicide residues in unpolished rice. The investigated herbicides were azimsulfuron, bensulfuron-methyl, chlorimuron-ethyl, chlorsulfuron, ethoxysulfuron, flazasulfuron, imazosulfuron, metsulfuron-methyl, pyrazosulfuron-ethyl and tribenuron-methyl. Acetonitrile water (2:1) extracts of rice samples were cleaned up with solid-phase extraction cartridges (octadecylsilane-bonded silica (ODS) and graphitized carbon black (GCB)). Three fractions of the GCB eluate were taken for analysis using 3 separate injections in order to avoid interference in LC-DAD analysis and to reduce analyte coelution problems. Recoveries from rice samples fortified with the 10 herbicides at 0.05 and 0.2 microgram/g ranged from 46.6 to 119.6%, and coefficients of variation were 3.1-12.6%. The quantitation limits were 0.01-0.02 microgram/g. PMID- 12092421 TI - [Pesticide regulation and residue monitoring in the United State of America]. PMID- 12092422 TI - [Fundamental knowledge of food allergy]. PMID- 12092424 TI - [Information guide for prion diseases]. PMID- 12092423 TI - [Mutagenicity test and their present status]. PMID- 12092425 TI - Taiwanese students' gender, age, interdependent and independent self-construal, and collective self-esteem as predictors of professional psychological help seeking attitudes. AB - Interdependent self-construal, collective self-esteem, age, and gender were used to predict attitudes toward seeking professional psychological help among a sample of junior high, high school, and college students in Taiwan (N = 594). Self-construal, collective self-esteem, and help-seeking attitudes were measured by the Self-Construal Scale (T. M. Singelis, 1994), the Collective Self-Esteem Scale Revised (R. Luhtanen & J. Crocker, 1992), and the Attitudes Towards Seeking Professional Psychological Help Scale (E. H. Fischer & J. L. Turner, 1970), respectively. By using stepwise regression, each of the independent factors with the exception of age significantly predicted the dependent variable, professional psychological help-seeking attitudes. Implications for counseling and future research are addressed. PMID- 12092426 TI - Mental health services for African Americans: a cultural/racial perspective. AB - African Americans have made consistent progress toward first-class citizenship since 1965. Nonetheless, mental health services for this population have been biased, incomplete, and deficient because similarities to European Americans have been emphasized whereas differences were largely ignored. This article addresses some differences, including cultural/racial identity and the cultural self, that affect assessment, psychiatric diagnoses, and psychotherapy or other interventions. An assessment-intervention model illustrates how cultural information can inform service delivery and improve services to African Americans. Cultural/racial perspectives on mental health in a multicultural society suggest innovations in the design of these services and access to them that can increase utilization by African Americans. PMID- 12092427 TI - Constructions of provider role identity among African American men: an exploratory study. AB - This exploratory study examined the identity constructions of African American men using a qualitative research methodology. Seven African American men, ranging in age from 20 to 47 years and whose education levels ranged from a 1st-year university student to a PhD, were interviewed for this study. Central to how all of these men defined themselves was the breadwinner or provider role. Participants emphasized education as "insurance" against discrimination and an awareness of educational and occupational opportunities. For these participants, education was a means of ensuring opportunity, which afforded fulfillment of the provider role. This study supports the work of N. Cazenave (1979, 1981), who demonstrated the salience of the provider role among African American men. The implications of the provider role among African American men for research are also discussed. The data also suggested diversity within the African American male experience. As 1 participant described African American men. "We come like flowers, you know. Some in bouquets, and some wild." By providing constructions of identity that diverge from existing negative stereotypes of African American men, this study attempted to deconstruct those stereotypes. Finally, this study provided a voice to an underrepresented group in the research literature. PMID- 12092428 TI - Psychological reactions to redress: diversity among Japanese Americans interned during World War II. AB - The psychological reactions of 2nd-generation (Nisei) Japanese Americans to receiving redress from the U.S. government for the injustices of their World War II internment were investigated. The respondents, all of whom had been interned during the war, rated the degree to which the receipt of redress nearly 50 years after their incarceration was associated with 8 different areas of personal impact. Results indicated that redress was reported to be most effective in increasing faith in the government and least effective in reducing physical suffering from the internment. Women and older respondents reported more positive redress effects. In addition, lower levels of current income, an attitudinal preference for Japanese Americans, and preredress support for seeking monetary compensation each increased the prediction of positive redress effects. Findings are discussed in relation to theories of social and retributive justice. PMID- 12092429 TI - Duplicate publication Part 1. Consideration of the issues. AB - Although duplicate publication is a significant problem for the nursing community, it can be prevented by teamwork between the author and editor. PMID- 12092432 TI - A patient remembers the miracle drug cortisone 50 years later. PMID- 12092431 TI - Revising a book chapter written by a previous author. AB - Throughout the process, if you have any questions or concerns, do not hesitate to contact the editor. Part of an editor's responsibility is to provide guidance and assistance to the contributing authors. PMID- 12092433 TI - Searching for a cure. PMID- 12092434 TI - Imprisoned by illness. PMID- 12092435 TI - Finding Amy Peterson. An olympic speed skater battles with chronic fatigue syndrome. PMID- 12092436 TI - Hmong shamanism. Animist spiritual healing in Minnesota. AB - Hmong cultural attitudes, values, and behaviors influence when, where, why, and with whom a Hmong person will use Western medicine. Understanding the practices and importance of Hmong healing traditions will help majority-culture physicians provide respectful and effective health care to Hmong patients. The foremost Hmong traditional healer is the shaman (tu txiv neeb, pronounced "too tse neng"). There is no equivalent health professional in Western biomedicine, and the scope of the shaman as a healer extends beyond the capacities and expertise of physicians. Despite 25 years of Hmong acculturation in the United States and conversion to Christianity, Hmong shamanism maintains its traditional role in health and healing. Many Hmong who see physicians also rely on shamans for restoring health and balance to their body and soul. Thus, the Hmong shaman can be considered a powerful complement to Western health care professionals. This article presents the results of semistructured interviews with 11 Hmong shamans (5 males, 6 females, ranging in age from 35 to 85) and 32 nonrandomly selected Hmong patients (14 males, 18 females ranging in age from 21 to 85). The shamans described their spiritual perspectives, training and skills, and professional activities. Patients described their beliefs about spiritual healing and health care. These interviews suggest that Shamanism is considered effective care by many Hmong, irrespective of age, gender, or degree of acculturation. The article also includes summary charts of Hmong healing practices and concludes with a set of questions designed to help practicing physicians access the assumptions and beliefs of their Hmong patients so that they can provide efficient, effective, and satisfactory care. PMID- 12092437 TI - Multiple sclerosis update. AB - Multiple sclerosis (MS) is the most common cause of neurological disability in young European and North American adults. It is defined as dysfunction in different parts of the central nervous system at different points in time. The etiology is likely autoimmune, resulting from environmental and genetic factors. This article summarizes the symptoms, progression, diagnosis, and treatment of MS. MRI and CSF studies have been extremely helpful in identifying individual cases, but the diagnosis remains a clinical diagnosis, and one of exclusion. Methylprednisolone remains the preferred treatment for acute relapses, although plasma exchange has been shown to be helpful in cases when methylprednisolone has failed. Preventive immunomodulatory treatments are the focus of many current research efforts. Although these treatments have been shown to reduce the frequency of MS attacks, the reduction is modest, at 30%. The data that these drugs alter the progression of disability is less compelling. PMID- 12092438 TI - Asthma in children. A report on the Minnesota Asthma Coalition. PMID- 12092439 TI - Diabetic retinopathy. The diabetes eye exam initiative. PMID- 12092440 TI - Back in the game. PMID- 12092441 TI - Women's oral health across the lifespan. Part One: Historical perspectives. AB - This two-part series is intended to provide dental practitioners with information related to the clinical aspects of women's health in general and women's oral health in particular. Part One in the series presents a historical perspective from the inception and evolution of women's oral health to current issues of relevance. Part Two in the series will present a developmental perspective on women's oral health across the lifespan from infancy through the post-menopausal stages. Emphasis will be focused on medical and dental aspects of each developmental stage. PMID- 12092442 TI - Practical magic: the new Hibbing Regional Dental Clinic. PMID- 12092443 TI - Mucocutaneous disorders--a guide for dental health care workers. AB - While only a few of the mucocutaneous disorders are presented here, many others exist. Often the dental team will be the first to observe clinical signs of mucocutaneous disorders, and therefore it is prudent to ask about eye, skin, and genital lesions, as patients may not often volunteer this information. In the case of Pemphigus, dentists can play a key role in early detection of a potentially fatal disorder. In cases where chronic gingival inflammation remains despite local measures and good patient oral hygiene, it may be wise to include a mucocutaneous disorder in the differential diagnosis. PMID- 12092444 TI - Techniques for root coverage. PMID- 12092445 TI - Sentinel Event Alert Advisory Group hits the ground running at first meeting. PMID- 12092446 TI - Managed care standards revisions approved for health care network program. PMID- 12092447 TI - Surveying for staffing effectiveness in hospitals. PMID- 12092448 TI - Gearing up to meet new staffing effectiveness requirements. PMID- 12092449 TI - A pictorial method of benchmarking. PMID- 12092450 TI - Immunology primer. AB - The immune system, through a complex interplay of highly specialized cells and a seemingly endless number of soluble mediators, works to ensure protection from the potentially harmful pathogens that we encounter in our lifetime. The development of the immune system is a compromise between the necessity to recognize foreign peptides in the context of self-molecules (MHC) and the need to be tolerant to all self-peptides. Despite the large number of mechanisms in place to ensure the removal or suppression of self-reactive lymphocytes, the system is not 100% effective, with the occasional result of autoimmunity. A number of autoimmune disorders occur together and a better understanding of the genetic basis underlying this association should lead to an enhanced ability to predict, diagnose, and develop therapies aimed at preventing and treating these diseases. PMID- 12092451 TI - Neonatal syndromes of polyendocrinopathy. AB - Fetal and newborn mammals have a limited ability to mount an immune response, both qualitatively and quantitatively, leading to an increased susceptibility to bacterial or viral infections. The incomplete development in the neonate of both innate and acquired immune system may well explain this increased susceptibility; however, the neonatal immune system, under certain circumstances, can mount an efficient immune response. Various immune-mediated disorders, including autoimmune syndromes, can take place in the early postnatal life, a period during which the immune system acquires key functions and undergoes a complex maturation and education process. Neonatal autoimmune syndromes involving endocrine glands include: IPEX/XLAAD/XPID, neonatal hyperthyroidism, Di George syndrome, ALPS, and Kabuki syndrome. PMID- 12092452 TI - APS-I/APECED: the clinical disease and therapy. AB - The clinical picture and course of APS-I or APD-I/APECED is widely variable: the list of possible disease components includes some 30 disorders. The initial manifestation may not include any of the known characteristic components, namely, mucocutaneous candidiasis, hypoparathyroidism, or adrenocortical insufficiency. Although mutation detection is available, it does not help to exclude this disease. Diagnostic strategy needs to be based on knowledge of the clinical picture, including the features of ectodermal dystrophy. PMID- 12092453 TI - Population genetics and functions of the autoimmune regulator (AIRE). AB - The autoimmune polyglandular syndrome type 1 (APS1), also known as autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APS1), is a monogenic autosomal disease with recessive inheritance. It is characterized by chronic mucocutaneous candidiasis, multiple autoimmune endocrinopathies, and ectodermal dystrophies. The defective gene responsible for this disease has been identified and named "autoimmune regulator" (AIRE). The AIRE gene is located on chromosome 21q22.3. At least 45 different disease-causing mutations in AIRE have been discovered. This review summarizes the global distribution of AIRE mutations and the relevance of major mutations to the clinical disorders associated with APS1. We also will review studies on the structure and DNA-binding ability of the AIRE protein and the possible malfunctions of the AIRE protein as a result of major disease-causing mutations. PMID- 12092454 TI - Autoimmune polyglandular syndrome. II: Clinical syndrome and treatment. AB - A high index of suspicion should be maintained whenever one organ-specific autoimmune disorder is diagnosed in order to prevent morbidity and mortality from the index disease as well as associated diseases. Further definition of susceptibility genes and autoantigens, and understanding of immune tolerance and the induction and propagation of autoimmune reactions should prove to be the best path to improved diagnostic and therapeutic modalities in the care of these patients. PMID- 12092455 TI - The genetics of autoimmune polyendocrine syndrome type II. AB - A series of autoimmune disorders, often Addison's disease, type 1 diabetes mellitus, and thyroid autoimmunity, frequently occurs together in patients with the autoimmune polyendocrine syndrome type II (APS-II). The highest risk HLA genotype for Addison's disease, either as a single disease or in APS-II patients, consists of the genotype DR3/4, DQ2/DQ8 with DRB1*0404. As many as 30% of patients with Addison's disease have this genotype versus less than 0.5% of controls. An additional and important associated locus within the HLA region is the class I related gene, MIC-A. Patients who develop Addison's disease often have a delayed diagnosis and may die from Addisonian crisis; therefore, improved genetic testing combined with testing for 21-hydroxylase autoantibodies might allow the identification of relatively high-risk populations (greater than 1 in 200 defined genetic risk compared with 1 in 10,000 population risk). PMID- 12092456 TI - Autoantibodies in autoimmune polyendocrine syndrome type II. AB - The autoimmune polyendocrine syndrome type II (APS-II) is characterized by the association of autoimmune Addison's disease with thyroid autoimmune diseases or type-1 diabetes mellitus. 21-Hydroxylase autoantibodies enable the accurate diagnosis of autoimmune Addison's disease and, in patients with other endocrine autoimmune diseases, identify subjects at high risk for clinical adrenal insufficiency. 17 alpha-Hydroxylase (17OH) and side-chain-cleavage enzyme (P450scc) are target autoantigens of steroid-cell autoantibodies, and in women with Addison's disease, 17OH autoantibodies and P450scc autoantibodies are markers of increased risk for premature ovarian failure. Thyroperoxidase autoantibodies, thyroglobulin autoantibodies, H+/K(+)-ATPase autoantibodies, and GAD65 autoantibodies are frequently detected in patients with isolated Addison's or APS-II. Screening for other organ-specific autoimmune diseases should be performed in every patient with at least one major disease component of APS-II. PMID- 12092458 TI - Peripheral antigen-expressing cells and autoimmunity. AB - There is growing evidence that self-molecules with tissue-restricted expression are also expressed at low levels in the thymus, where such expression may affect the development of self-tolerance. Genetic factors may modulate such expression and, in turn, influence susceptibility to autoimmune responses against specific molecules. Contrasting reports exist regarding the phenotype of the cells that express self-molecules in the thymus. Indeed, both bone marrow derived antigen presenting cells and thymic epithelial cells were reported to express self molecules with tissue-restricted expression. Further studies of these cells and the mechanisms by which they exert their putative tolerogenic effects clearly are necessary. PMID- 12092457 TI - Type 1A diabetes mellitus-associated autoimmunity. AB - Type 1A diabetes mellitus has become one of the most intensively studied autoimmune disorders, with characterized animal models and extensive prospective studies of the development of anti-islet autoimmunity. It is now possible to predict the development of type 1A diabetes mellitus, beginning with HLA-encoded genetic susceptibility, followed by the development of a series of anti-islet autoantibodies. Prediction primarily is based on the detection of multiple anti islet autoantibodies reacting with cloned islet antigens. Multiple international workshops fostered the development of specific and sensitive radioassays for autoantibodies reacting with GAD65 (glutamic acid decarboxylase), ICA512 (also termed IA-2, a tyrosine phosphatase-like protein), and insulin. Similar high throughput radioassays have been applied using autoantigens for additional autoimmune disorders including celiac disease and Addison's disease. Relatives of patients with type 1A diabetes mellitus inherit susceptibility to express multiple autoantibodies, and a subset of autoantibody-positive individuals inherit susceptibility to progress to overt disease. This article reviews autoimmune disorders associated with type 1A diabetes mellitus. PMID- 12092459 TI - Animal models of autoimmune polyglandular syndrome. AB - Animal models for autoimmune polyglandular syndrome that are either spontaneous or induced by manipulations are presented. In addition, recent cloning of the autoimmune regulator (AIRE) gene responsible for autoimmune polyglandular syndrome type I, and its homologue in mice, Aire, provided the opportunity to study the effect of this gene on autoimmune diseases by producing knockout mice. These animal models make it possible to perform experiments that cannot be performed in humans, which will increase our understanding of the cause and molecular mechanisms of the autoimmune diseases and lead to the development of effective methods for their prevention and intervention. PMID- 12092460 TI - Immunomodulatory vaccination in autoimmune disease. AB - The development of vaccines is arguably the most significant achievement in medicine to date. The practice of innoculation with the fluid from a sore to protect from a disease actually dates back to ancient China; however, with the introduction of Jenner's smallpox vaccine, and greater understanding of the immune system, vaccines have become specific and systematic. Traditional vaccines have used killed pathogens (hepatitis A and the Salk polio vaccines), immunogenic subunits of a given pathogen (hepatitis B subunit vaccine), or live attenuated pathogens (measles, mumps, rubella, Sabin polio vaccines) to generate protective immunity. Currently, a new generation of vaccines that use the genetic material of a pathogen to elicit protective immunity are being developed. Although the most widespread and successful use of vaccines today remains in the arena of infectious diseases, manipulations of immune responses to protect against cancers, neurologic diseases, and autoimmunity are being explored rigorously. PMID- 12092461 TI - The use of monoclonal antibodies to restore self-tolerance in established autoimmunity. AB - The author hopes to convince the reader that the data presented argue for a stage during the development of IDDM when beta-cell destruction can be counteracted and tolerance to beta cells restored, provided the immune aggression is arrested. This argument constitutes a solid rationale for immunointervention in established IDDM, especially by using potent agents such as CD3. The future for the application of monoclonal antibodies not only in autoimmunity but also in transplantation is exiting. With the development of humanized monoclonal antibodies, therapeutic uses for them are likely to expand. Enormous progress has been made in the last 15 years, and it is likely that before a similar time period has elapsed, monoclonal antibodies will have become standard tools that will dispense the need for long-term immunosuppression and its inherent dangers in various clinical arenas. PMID- 12092462 TI - Cytokine and immunosuppressive therapies of type 1 diabetes mellitus. AB - In this article, the authors covered a number of issues that affect how researchers approach prevention of diabetes. The focus has been the use of cytokines and immunosuppressive therapies. The historical understanding of cytokine and immunosuppressive approaches, new developments in using these agents in humans, and the issues involved in designing diabetes prevention trials were reviewed. Although progress at times appears slow, the current research activities predict new developments in the next few years that may improve the understanding of the progression of diabetes and possible ways to intervene. PMID- 12092463 TI - [A study of the impact of the introduction of a vaccine into the immunization calendar]. PMID- 12092464 TI - [The evaluation of effectiveness of scholastic programs in the prevention of consumption of tobacco, alcohol and cannabis: what do meta-analyses tell us?]. AB - Consumption of tobacco, alcohol and illegal drugs is a major public health problem in developed countries. The aim of the study is to describe the impact and associated characteristics of preventive programs addressed at those problems in the school setting. Meta-analysis focusing on evaluations of programs focusing on smoking, alcohol and/or cannabis at the school setting are reviewed. The search was done at Cochrane Library and Medline databases of articles published between 1993 and 1999, and including as keywords programs, education, drugs prevention, prevention, smoking, alcohol, school, adolescence, teenagers, young people, evaluation, health education, effectiveness, review, meta-analysis. We found 5 meta-analysis of programs summarizing the effect of preventive programs, most of them dealing with legal and illegal drugs. However, most of the interventions reporting changes in behavior measured only smoking. More effective interventions addressed social influences, used active methodology and were implemented by teachers or peers. The importance of booster sessions, the quality of implementation and thorough evaluation is stressed. Overall, meta-analysis of evaluated programs shows a small effect, although the population impact may be relevant. Some limitations point to new areas of interest for future research. PMID- 12092465 TI - [Advances of the investigation of "Ambulatory Care Sensitive Conditions" in primary care in Spain]. AB - Hospitalization due to ambulatory care sensitive conditions (ACSC) is an indicator of hospital activity that has demonstrated its usefulness as an indirect measurement of primary care effectiveness. Since this indicator was recently introduced in Spain, a collaborative effort between the different research groups could facilitate and promote its development and progress. The objective of this paper is to propose a working agenda that, starting from the most recent information, enhances the advance in this research field. The agenda includes the following sections: 1) To draw up specific ACSC lists for adult and pediatric population, as well as to look in greater depth into the concepts of, and differences in avoidable hospitalization and ACSC. 2) To complete the indicator validation process by assessing the external validity. 3) To propose, for future studies, the municipality as the unit of analysis, as well as to individualize the analysis of health conditions allowing for the differences between acute and chronic ones. 4) To adjust the indicators of hospital activity by hospital use index, when data from some hospitals are lacking and comparisons are wanted 5) To include a new variable, provider of primary health care services, in the Minimum Basic Data Set of Hospital Discharges. 6) To use this indicator as a measure of both the distribution of functions between levels of care and the coordination among them. PMID- 12092466 TI - [Haemophilus Influenzae infection before and after the immunization campaign among children in the Valencia community (1996-2000)]. AB - BACKGROUND: The introduction of a conjugate type b Haemophilus influenzae (Hib) vaccine for children has led to a sharp drop in the incidence of H. influenzae disease. The purpose of this study is that of analyzing the major characteristics of invasive disease due to H. influenzae as regards epidemiology, clinical aspects, evolution and immunization status among the infantile population of the Autonomous Community of Valencia for the 1996-2000 period. METHOD: The data was taken from the clinical records of those children under age 15, who have shown clinical signs and symptoms indicative of invasive disease entailing isolation of Haemophilus influenzae and/or meeting the established case definition requirements, who were treated at all of the public hospitals in the Autonomous Community of Valencia throughout the 1996-2000 period. The trend of incidence was assessed by mean of incidence rates. The clinical pattern and the evolution there of (sequelae and life-threatening aspects) by frequency and age range. RESULTS: A total of 36 cases of invasive disease due to Haemophilus influenzae were recorded. The incidence rate among children under age 15 dropped from 3.56/10(5) in 1996 to 1.07/10(5) in 1997 (coinciding with the immunization campaign and the subsequent including of the conjugate Hib vaccine in the Routine Vaccination Schedule of the Autonomous Community of Valencia) and 0.30/10(5) in 1998, this being a situation which has continued over the following years. Fifty-three percent (53%) of the cases occur in children under age 18 months. Both the sequelae as well as the deaths occurred throughout the period prior to the routine use of the conjugate vaccine. No child properly immunized died. Two cases of non-b type H. influenzae occurred in immunized children. CONCLUSIONS: The incidence of infection due to type b Haemophilus influenzae dropped drastically as of the start of the routine immunization of the infantile population. PMID- 12092467 TI - [Characteristics of antibiotic prescription during office visits in the Hospital Cayetano Heredia in Lima, Peru]. AB - BACKGROUND: The present descriptive cross-sectional study, assesses the proportion of antibiotic prescription (PPA), and the quality of it (CPA) in the ambulatory consultation of the department of medicine at the Cayetano Heredia Hospital, as an approach to antibiotic use in Peruvian hospitals. METHODS: The study population was conformed by the patients who went to outpatient care from January 8th to February 2nd, 2001; a sample size of 120 patients was determined out of 1449 patients, to assess CPA. Data on antibiotic prescription was collected, revised and qualified by three experienced physicians, using as gold standard the United States Pharmacopeial Drug Information criteria. RESULTS: PPA was 13.53% (CI 95%: 11.77%-15.29%), without significant differences according to age, sex, doctor's offices and turns. The most frequent diagnoses with antibiotic prescription were: urinary tract infection and pharyngoamigdalitis; the antibiotics more frequently prescribed were: ciprofloxacine and cotrimoxazol. Of the antibiotics, 70.00% were prescribed as non commercial name and 70.83% were registered in the National Petitory of Essential Drugs. We found that 81.67% (CI 95%: 79.68%-83.66%) of the prescriptions were inadequate mainly due to: duration (59.20%) and dose (20.00%) of the antibiotic treatment. CONCLUSIONS: PPA obtained is lower than those published in, and CPA is as high as the reported one in other studies. The investigations in this field are still scarce. It is important new research in order to know about prescription, use and adverse reactions of antibiotics. It is necessary to develop a antibiotic policy to improve access and rational use of antibiotic. PMID- 12092468 TI - [Dependence and necessities of non-covered services for the elderly in a health sector of Zaragoza]. AB - BACKGROUND: Dependency to carry out the activities of daily living (ADL) and home care needs of older adults depend upon factors which are not well understood. Caregiving is mainly provided by the family. The aim of this study is to estimate the prevalence of ADL dependence and associated factors among older adults and to identify associated factors with unmet home care needs in people over age 75 in Zaragoza (Spain). METHOD: A cross-sectional study was conducted on a representative sample of the community dwelling population over age, 75 in one Health Care District in Saragossa in 1998 (n = 351). Data were collected during a personal interview. ADL dependency was assessed by the Katz index. Home care needs were assessed using prevalent norms for desired frequency of care. Sources of informal care, socio-demographic characteristics, self-rated health and depression (Yesavage Geriatric Depression Scale) were considered independent variables. Logistic regression was used to identify factors associated with both ADL dependency and unmet home care needs. RESULTS: Prevalence of ADL dependence is 37.3%. The variables predicting ADL dependence are the perception of poor health, depression, being a female and being over 85 years of age. The prevalence of unmet care needs among ADL dependent individuals is 22.1%. Sources of help are limited to the families. Being a woman and living alone are the stronger risk factors for unmet needs. CONCLUSIONS: A high percentage of older adults are dependent. Informal support is insufficient and confined to the family. The social and health care system in Spain should provide to the family, emotional, financial and social support and special care for disabled people living alone and with limited material resources. PMID- 12092469 TI - [Predictors of the onset of tobacco consumption among secondary school students of Barcelona and Lleida]. AB - BACKGROUND: Knowing the factors predisposing the smoking onset of smoking at school age may help to improve the preventive strategies and make the starting age later. This study is set out to describe the natural history of smoking throughout the teen-age years and the main factors which may predict teenagers developing the smoking habit. METHODS: This is a longitudinal study of a survey type, using the school room as the sampling unit at six schools in Barcelona and Lleida. A total of 1,460 students within the 12-19 age range took part. Two surveys were conducted, an initial survey in the spring of 1992 and a follow-up survey one year later. A total of 1,236 questionnaires (84.3%) were paired by means of a personal code used for the longitudinal study. RESULTS: A total of 25% of the students were regular smokers at the start of the study, 17.1% smoking occasionally or having experimented with smoking, there being no differences between sexes for both of these categories. Among those who were non-smokers at the starting survey, 12.7% stated that they smoked occasionally and 6.2% regularly one year later. The females started smoking at an earlier age, the differences as compared to males being statistically significant at 14 years of age. The predictors of the smoking onset include favorable attitudes toward cigarettes (OR = 3.1; CI5% (1.6-5.6), drinking alcoholic beverages (OR = 2.2; CI95% (1.4-3.3), being younger than age 15 (OR = 2.0; CI95% (1.2-3.4), having friends who smoke (OR = 1.9, CI95% (1.2-3.0) and the stated intention of smoking in the future (OR = 1.6; CI95% (1.0-2.5). CONCLUSIONS: The students showing a higher degree of tolerance toward cigarette smoking, entailing favorable attitudes or friends who smoke or who intend to smoke in the future at the start of the study are at a much higher risk of Smoking onset than their classmates who are not of these characteristics. The cigarette smoking prevention programs must focus special attention on the teenagers most highly vulnerable to these factors. PMID- 12092470 TI - [The incidence of respiratory tuberculosis in the province of Leon according to the system of notification of disease mandatory declaration, 1992-1999]. AB - BACKGROUND: Tuberculosis continues to be a major public health problem. Knowing the incidence, trend and geographical spread of reported tuberculosis may serve to evaluate the prevention and control measures implemented and point to further needs. METHODS: Retrospective study of the reported cases of tuberculosis in the province of Leon, broken down by years, Health Care Areas and Basic Health Care Districts for the 1992-1999 period. RESULTS: The average annual incidence rates were 40.3, 38.6 and 44.4 per 100,000 inhabitants/year (10(5) inhabitants/year) for the province of Leon, Leon Health Care Area and Bierzo Health Care Area, respectively. Seventy percent (70%) were males and thirty percent (30%) females. The specific rates by sex in the province of Leon were 55.5 for males and 22.9 per 10(5) for females. The specific rates by age and sex were similar for males and females up to age 25, the incidence among males being much greater as of this age. The geographical spread of tuberculosis in the province of Leon was heterogeneous, the highest incidence being in mining areas. In the Leon Health Care Area, cases among the oldest adults prevailed, no differences being found among young adults and adults over age 55 in the Basic Health Care District. CONCLUSIONS: The province of Leon has an incidence of tuberculosis greater than that estimated for Spain as a whole. It would be necessary to step up epidemiological surveillance and control measure systems as directly-monitored cases detected and systematic study of contacts in order to achieve a greater efficiency regarding the control of this disease. PMID- 12092471 TI - [Prevalence of anemia due to the iron deficiency among children between 6 months and 5 years of age in the municipal area of Arismendi of the Nueva Esparta State. Venezuela 2001]. PMID- 12092472 TI - Progress toward a global understanding of infective endocarditis. Early lessons from the International Collaboration on Endocarditis investigation. AB - In the century and a quarter since William Osler delivered his framed lectures on endocarditis substantial advancements have occurred in the understanding and treatment of this disease. This article summarizes current understanding of endocarditis in the areas of population epidemiology, host factors, microorganisms, diagnosis, and therapy. In addition, the authors discuss possible directions for investigation in the future, including a new multinational consortium, the International Collaboration on Endocarditis (ICE). This collaboration aims to provide a mechanism to advance the understanding of endocarditis in areas difficult to study without an established network. The multinational nature of the collaboration may also permit a more global view of IE and provide opportunities for studies such as randomized trials of therapeutic treatment strategies. PMID- 12092474 TI - Pathogenesis of streptococcal and staphylococcal endocarditis. AB - Although streptococcal and S. aureus IE share the same primary site of infection, their pathogenesis and clinical evolution present several major differences. Streptococci adhere to cardiac valves with pre-existing endothelial lesions. In contrast, S. aureus can colonize either damaged endothelium or invade physically intact endothelial cells. These interactions are mediated by multiple surface adhesins, some of which have been only partially characterized. Streptococci produce surface glucans (gtf and ftf), ECM adhesins (e.g., fibronectin-binding proteins, FimA), and platelet aggregating factors (phase I and phase II antigens, pblA, pblB, and pblT), all of which have been. PMID- 12092473 TI - Infective endocarditis in intravenous drug abusers and HIV-1 infected patients. AB - Infective endocarditis (IE) is one of the most severe complications of parenteral drug abuse. The incidence of IE in intravenous drug abusers (IVDAs) is 2% to 5% per year, being responsible for 5% to 20% of hospital admissions and 5% to 10% of the overall death rate. IVDAs often develop recurrent IE. The prevalence of HIV infection among IVDAs with IE ranges between 30% and 70% in urban areas in developed countries. The incidence of IE in IVDAs is currently decreasing in some geographical areas, probably due to changes in drug administration habits undertaken by addicts in order to avoid HIV transmission. Overall, Staphylococcus aureus is the most common etiological agent, being in most geographical areas sensitive to methicillin (MSSA). The remainder of cases is caused by streptocococci, enterococci, GNR, Candida spp, and other less common organisms. Polymicrobial infection occurs in 2% to 5% of cases. The tricuspid valve is the most frequently affected (60% to 70%), followed by the mitral and aortic valves (20% to 30%); pulmonic valve infection is rare (< 1%). More than one valve is infected in 5% to 10% of cases. HIV-positive IVDAs have a higher ratio of right sided IE and S. aureus IE than HIV-negative IVDAs. Response to antibiotic therapy is similar among HIV-infected or non-HIV-infected IVDAs. Drug addicts with non complicated MSSA right-sided IE can be treated successfully with an i.v. short course regimen of nafcillin or cloxacillin for 2 weeks, with or without addition of an aminoglycoside during the first 3 to 7 days. Surgery in HIV-infected IVDAs with IE does not worsen the prognosis. The prognosis of right-sided endocarditis is generally good; overall mortality is less than 5%, and with surgery less than 2%. In contrast, the prognosis of left-sided IE is less favorable; mortality is 20% to 30%, and even with surgery is 15% to 25%. IE caused by GNB or fungi has the worst prognosis. Mortality between HIV-infected or non-HIV-infected IVDAs with IE is similar. However, among HIV-infected IVDAs, mortality is significantly higher in those who are most severely immunosuppressed, with CD4+ cell count < 200/microL or with AIDS criteria. Finally, IE in HIV-infected patients who are not drug abusers is rare. PMID- 12092475 TI - Imaging techniques for diagnosis of infective endocarditis. AB - With the ability to structurally characterize cardiac manifestations, echocardiography is used for the diagnosis and management of infective endocarditis. In establishing the diagnosis according to the Duke criteria, the findings of endocardial involvement (vegetation, abscess, prosthetic valve dehiscence) or new valvular regurgitation represent "major" diagnostic criteria. As echocardiography cannot reliably differentiate noninfective from infective lesions, however, proper diagnosis lies in correlating echocardiography with clinical findings. The more invasive transesophageal approach provides substantially greater image resolution; this approach should be considered first in the evaluation of patients with higher prior probabilities of endocarditis and those with potential endocardial complications. PMID- 12092476 TI - Diagnostic methods current best practices and guidelines for histologic evaluation in infective endocarditis. AB - Infective endocarditis (IE) often presents diagnostic and therapeutic challenges and continues to cause high morbidity and mortality. Confirmation of the diagnosis of IE is important for the purposes of epidemiologic and clinical studies and is crucial for patient management. Despite recent advances in diagnostic techniques, about 10% of IE cases remain culture-negative. Because pathological examination of cardiac valves to demonstrate vegetations and valvular inflammation remains the gold standard for the diagnosis of IE, the role of the pathologist is often decisive, especially when bacteriologists fail to isolate a microorganism or when a microorganism that has been isolated may be a contaminant. Furthermore, the pathologist may play an important role in identification of previously unknown infectious agents. PMID- 12092477 TI - Diagnostic methods current best practices and guidelines for isolation of bacteria and fungi in infective endocarditis. AB - This article addresses the clinical need for accurate detection of bacteria and fungi that may cause infective endocarditis (IE). The pathophysiology of bacteremia during endocarditis is reviewed to provide an understanding of how current diagnostic methods may be used to determine the etiologic diagnosis. The critical contribution of microbiologic findings to the current clinical diagnostic criteria is emphasized. Finally, "best practices" recommendations are presented to optimize the likelihood of reaching a correct etiologic diagnosis while minimizing the number of "culture-negative" cases. PMID- 12092478 TI - Diagnostic methods current best practices and guidelines for identification of difficult-to-culture pathogens in infective endocarditis. AB - Culture-negative endocarditis currently represents a diagnostic challenge for physicians. Traditional methods such as histology, serology, and culture have been improved and new molecular techniques have been developed to improve the detection of difficult-to-culture agents. Serologic tests for the two most frequent etiologic agents, Coxiella burnetii and Bartonella spp, should be performed first because they can usually be identified easily in this way. The sensitivity of culture for intracellular bacteria has been improved by inoculation of samples in shell vials and by the use of novel tissue cell lines. Recently, universal and species-specific primers have been designated to amplify bacterial DNA directly from resected valves, allowing positive identification. PMID- 12092480 TI - Staphylococcus aureus bacteremia and endocarditis. AB - Staphylococcus aureus is a leading cause of bacteremia and endocarditis. Over the past several years, the frequency of S. aureus bacteremia (SAB) has increased dramatically. This increasing frequency, coupled with increasing rates of antibiotic resistance, has renewed interest in this serious, common infection. S. aureus is a unique pathogen because of its virulent properties, its protean manifestations, and its ability to cause endocarditis on architecturally normal cardiac valves. Although the possibility of underlying endocarditis arises in virtually every patient with SAB, only a minority of bacteremic patients will actually have cardiac involvement. Distinguishing patients with S. aureus infective endocarditis (IE) from those with uncomplicated SAB is essential, but often difficult. In this review, the authors summarize recent changes in the epidemiology of SAB and IE, discuss the challenges in distinguishing SAB from IE, and discuss current trends in the management of patients with SAB and IE. PMID- 12092481 TI - Special issues in the management of infective endocarditis caused by gram positive cocci. AB - Gram-positive cocci, mainly streptococci and staphylococci, continue to cause the majority of cases of infective endocarditis. Among the streptococci causing IE, the long-standing predominance of oral or viridans-group streptococci has progressively faded, while the number of cases caused by "enteric streptococci" (Streptococcus bovis and enterococci) has increased. While most oral streptococci and S. bovis strains remain fully sensitive to penicillin, nutritionally variant streptococci--now renamed Abiotrophia--and enterococci can exhibit resistance to penicillin and/or glycopeptides that makes endocarditis more difficult to treat. Among the staphylococci causing endocarditis, the increasing proportion of coagulase-negative and methicillin-resistant strains observed in recent years has changed the approach to choice of antibiotic therapy. The purpose of this paper is to focus on some new aspects of the management of antibiotic therapy of IE due to streptococci and staphylococci, including recent developments such as once daily aminoglycoside administration in IE, outpatient antibiotic therapy, and the evaluation of new antibiotics. PMID- 12092479 TI - Molecular methods for diagnosis of infective endocarditis. AB - The culture of viable microorganisms from the blood or from cardiac tissue is currently the most important test for diagnosis of IE. This is followed by phenotypic identification methods used for taxonomic positioning of isolates. However, in those cases where the invading microorganism is difficult or impossible to culture (including instances of prior antimicrobial treatment), molecular methods provide the best means for detection. Molecular identification methods, either nucleic acid target or signal amplification alone or in combination with sequence analysis can offer a more specific and in some cases a more rapid alternative to the phenotypic methods. We propose revised Duke criteria of IE, including positive identification of an organism by molecular biology methods. PMID- 12092482 TI - Current best practices and guidelines indications for surgical intervention in infective endocarditis. AB - Optimal diagnosis and management of patients with infective endocarditis requires sound clinical judgment based on extensive experience. This is especially important in regard to the indications and timing for surgery. To achieve the best possible outcomes, surgical intervention during treatment is required in 25% to 30% of patients with infective endocarditis. Heart failure and progressive left-sided valvular dysfunction are the most common indications for operation. Valve repair should be considered as an alternative to valve replacement whenever feasible, especially in younger patients. Successful management of perivalvular abscesses and prosthetic valve infections requires radical removal of infected tissue followed by reconstructive procedures performed by experienced surgeons. Emergency or urgent surgery should seldom be delayed. PMID- 12092483 TI - Infections of intracardiac devices. AB - Intracardiac devices are increasingly used to correct hemodynamically dysfunctional valves and electrophysiologic abnormalities. These devices become infected at relatively low rates. Nevertheless, when these low rates are applied to widely used devices, significant numbers of infections result. Additionally, these infections have been associated with high degrees of morbidity and high mortality rates. This article reviews the epidemiology, microbiology, clinical presentation, and medical as well as surgical therapy of intracardiac device infections. PMID- 12092484 TI - Current best practices and guidelines. Assessment and management of complications in infective endocarditis. AB - The most important complications of endocarditis are congestive heart failure, paravalvular abscess formation, and embolism, especially stroke. In addition, endocarditis may be complicated by septic arthritis, vertebral osteomyelitis, pericarditis, metastatic abscesses and an array of renal problems ranging from immune-complex glomerulonephritis to renal abscesses. Adverse reactions associated with medical treatment of endocarditis can also result in significant complications such as ototoxicity and nephrotoxicity, skin rashes, and serum sickness. This review focuses on the cardiac, embolic, neurologic and renal complications of endocarditis and discusses how these complications influence the clinical management of individual cases in daily practice. PMID- 12092485 TI - Responsibilities of scientists and engineers: theory and practice. PMID- 12092486 TI - Changes of climate in the development of practical ethics. PMID- 12092487 TI - Trading jobs for health: ionizing radiation, occupational ethics, and the welfare argument. AB - Blue-collar workers throughout the world generally face higher levels of pollution than the public and are unable to control many health risks that employers impose on them. Economists tend to justify these risky workplaces on the grounds of the compensating wage differential (CWD). The CWD, or hazard-pay premium, is the alleged increment in wages, all things being equal, that workers in hazardous environments receive. According to this theory, employees trade safety for money on the job market, even though they realize some of them will bear the health consequences of their employment in a risky occupational environment. To determine whether the CWD or hazard-pay premium succeeds in justifying alleged environmental injustices in the workplace, this essay (1) surveys the general theory behind the "compensating wage differential"; (2) presents and evaluates the "welfare argument" for the CWD; (3) offers several reasons for rejecting the CWD, as a proposed rationale for allowing apparent environmental injustice in the workplace; and (4) applies the welfare argument to an empirical case, that of US nuclear workers. The essay concludes that this argument fails to provide a justification for the apparent environmental injustice faced by the 600,000 US workers who have labored in government nuclear weapons plants and laboratories. PMID- 12092488 TI - Responsible authorship and peer review. AB - In this article the basic principles of responsible authorship and peer review are surveyed, with special emphasis on (a) guidelines for refereeing archival journal articles and proposals; and (b) how these guidelines should be taken into account at all stages of writing. PMID- 12092489 TI - Coauthorship in physics. AB - In a large and detailed survey on the ethics of scientific coauthorship, members of the American Physical Society (APS) were asked to judge the number of appropriate coauthors on his or her last published paper. Results show that the first or second coauthors are more appropriate than later coauthors about whom there is equal and considerable doubt. The probability of any third and subsequent coauthors being judged as inappropriate is 23% for the APS guideline, 67% for the tighter guideline of the International Committee of Medical Journal Editors, 59% if the guideline requires "direct contributions to scientific discovery or invention". Only 3% of respondents report having personally rejected an undeserving scientist who expected to be an author on the last published paper. Respondents seem to be divided into two non-overlapping populations--those who report no inappropriate coauthorship and those who have a more graduated view. PMID- 12092490 TI - Six domains of research ethics. A heuristic framework for the responsible conduct of research. AB - The purpose of this paper is to provide a simple yet comprehensive organizing scheme for the responsible conduct of research (RCR). The heuristic offered here should prove helpful in research ethics education, where the many and heterogeneous elements of RCR can be bewildering, as well as research into research integrity and efforts to form RCR policy and regulations. The six domains are scientific integrity, collegiality, protection of human subjects, animal welfare, institutional integrity, and social responsibility. PMID- 12092491 TI - Teaching social responsibility: the Manhattan project. Commentary on "The Six Domains of Research". AB - This paper discusses the critical necessity of teaching students about the social and ethical responsibilities of scientists. Both a university scientist and a middle school science teacher reflect on the value of teaching the ethical issues that confront scientists. In the development of the atomic bomb in the US-led Manhattan Project, scientists faced the growing threat of atomic bombs by the Germans and Japanese and the ethical issues involved in successfully completing such a destructive weapon. The Manhattan Project is a prime example of the types of ethical dilemmas and social responsibilities that scientists may confront. PMID- 12092492 TI - Four observations about "Six Domains of Research Ethics". AB - Stimulated by Kenneth Pimple's "Six Domains of Research Ethic", this paper examines four aspects of the responsible conduct of research and scientists' social responsibilities. I argue that scholars and decision-makers concerned with the responsible conduct of research should take notice of the rapidly growing body of scholarship on the social organization of science and the behavior of scientists, integrating that work with ethical principles. Of particular concern are the increasing heterogeneity and interdisciplinary of research, the ambivalences in the practice of peer review, the social tensions of research life, and the heightened concern for social and economic returns from federal research investments. In all, the paper echoes and develops Pimple's call for integrative thinking about the responsible conduct of research. PMID- 12092493 TI - Ethical decision-making in research: identifying all competing interests. Commentary on "Six Domains of Research Ethics". AB - Ethical dilemmas are the result of conflicts between potential benefits or harms for two or more competing interests. Therefore, ethical decision-making implies a responsibility to identify those interests, harms, and benefits. For this purpose, researchers have responsibilities to the research, the subjects of research, other researchers, the institution, society, the environment, and self. PMID- 12092494 TI - Making sense of scientists' responsibilities at the interface of science and society. Commentary on "Six Domains of Research Ethics". AB - As Kenneth Pimple points out, scientists' responsibilities to the larger society have received less attention than ethical issues internal to the practice of science. Yet scientists and specialists who study science have begun to provide analyses of the foundations and scope of scientists' responsibilities to society. An account of contributions from Kristen Shrader-Frechette, Melanie Leitner, Ullica Segerstrale, John Ahearne, Helen Longino, and Carl Cranor offers work on scientists' social responsibilities upon which to build. PMID- 12092495 TI - Beyond fabrication and plagiarism: the little murders of everyday science. Commentary on "Six Domains of Research Ethics". AB - Much of the focus of programs designed to promote responsible conduct in research has traditionally been on the high crimes of fabrication, falsification, and plagiarism. We believe that equally deserving of our attention are the misdemeanors that also can occur. Viewed as individual events, these "little murders" are far less serious. Yet, we believe that in the aggregate they can do great harm, not the least because they can set the stage for far greater crimes. PMID- 12092496 TI - Reflections on 'Real Science: What it is, and what it means' by John Ziman. AB - In these reflections on the recent book by John Ziman entitled 'Real Science: What it is and what it means', I have sought to review his main points and carry on the discussion that Ziman seeks to provoke. His approach to this subject arises from what exists on the ground and the way practising scientists view this area. I have taken a wider more abstract view of what is entailed by science than Ziman and have examined the implications of that choice. As we are presently being urged to effect more and more cross and multi-disciplinary activities and research, the boundaries that may be used to set aside that which is scientific are exposed to additional strains that challenge their integrity. The continuation of this discussion into the future is clearly called for; its outcome is less apparent. PMID- 12092497 TI - [Histology of the larynx]. AB - The mucous membrane lining the interior of the larynx was studied in autopsy specimens taken from 21 infant larynxes, with ages ranging from 1 to 15 years. Only those larynxes in which the epithelial mucosal structures were preserved were set aside for study. A standard histological technique was applied. Paraffin sections were stained with haematoxylin-eosin, PAS, Masson's triple stain and orcine. The epithelium of the ventricles, ventricular bands and subglottis was found to be of respiratory type. On the vocal folds, the lining was non keratinising squamous epithelium. These findings are consistent with those found in the literature. Findings that differed were as follows: the laryngeal surface of the epiglottis is classically described as having a respiratory type of epithelium, whereas we found it to be of non-keratinising stratified squamous type. The literature describes the epithelium of the ventricular bands as respiratory. While in our series we found stratified squamous in 56% and respiratory in 44%. Other findings are imprecise in the literature, and need definition; the finding of taste papillae in six cases without age correlation; and the richness of follicles in the lymphoid tissue at the epiglottis and ventricles of children aged more than 3 months. PMID- 12092498 TI - [Results of six years experience using an injection of autologous fat into the vocal cord]. AB - Several techniques are available for medialising the vocal cord or for compensating for glottic air escape. The authors have used the technique of injection of autologous fat into the vocal cord over a six year period for various indications, in a total of 124 patients (65 having unilateral palsy, and 59 glottic air escape). The later results depend on the initial problem, but in general demonstrate a notable reduction in air escape. PMID- 12092499 TI - ["Bamboo nodes": a clinical study of 19 cases]. AB - This is an account of 19 cases undergoing surgery for submucosal deposits on the cords of a particular type, 8 of the 19 cases having an auto-immune disorder. The nineteen patients were operated and followed up between December 1986 and December 2001, the main symptom being dysphonia. Stroboscopy demonstrated a yellowish appearance, often transverse, of the middle third of the cord, giving the characteristic appearance of a bamboo node. Suspension microlayngoscopy was performed in all cases. Cordotomy was invariably required. Examination of the case notes of these patients has shown either a pre-existing auto-immune disorder or the development of suggestive pointers in 8 of these female patients. Histological examination necessarily demonstrates the common characteristics of these deposits, and may be able to show features which have predictive value in the absence of known auto-immune pathology. PMID- 12092500 TI - [Life after total laryngectomy]. AB - This work relates the observation conducted in our outpatient clinic with fourteen patients who underwent a total laryngectomy due to a laryngeal and hypo pharyngeal cancer. The aims of our study were to analyse identity transformations and sociability modifications for these patients. It confirmed the social inequalities when a cancer occurs. Total laryngectomy upset or staggers the patients by impairing their appearance. Relations with other people are limited to the dearest because the tracheostomy and the deprivation of the original voice are regarded as disability that excludes them from the society. The physicians should be aware of the profound psychologic trouble and social changes attached to laryngeal cancer surgery to allow optimal care for the patients. PMID- 12092501 TI - [A protocol for the evaluation of the voice and speech of the hearing impaired]. AB - This study provides a methodology for the evaluation of the voice and the speech of deaf people (VANFIBR). This protocol uses a subjective multi parameter method, similar as the one used for the "normal-hearing" voice (GIRBAS). We use a global appreciation factor (A), a factor for the control of the pitch of the voice (F), the intensity (I), the extra-vocal noises (B), the rhythm of the speech (R) and the voisement (V). This method wants to be independent of the deafness pathology and the age of the subject. It was tested on a deaf population and a normal hearing one. The analyses by a jury showed the good discrimination between both populations and the pertinence of most of the parameters used. PMID- 12092502 TI - [Objective analysis of postural tonus using Fukuda's test. An attempt at correlation with vocal cord lesions]. AB - The authors summarise a study carried out in the University Hospital of Liege which attempts to correlate vocal cord pathology, posture, and the concept of muscular chains. They have hoped to develop the concept that a postural disorder may be the origin of demonstrable asymmetrical vocal cord disorders. Could there be a bilateral causal relationship between postural disorder and a predominately vocal pathology? How could more precise and long-lasting treatment be instituted? PMID- 12092503 TI - [The place of speech therapy in the dysfunctional dysphonias with gastro esophageal reflux]. AB - This work deals with the association between dysfunctional dysphonia and gastro oesophageal reflux. Joint medical and speech therapy management, adapted daily, would seem to be necessary. The authors report the results of a clinical assessment by questionnaire in six patients. PMID- 12092504 TI - [Is it possible to evolve the strategic approach to phonation problems?]. AB - The suggested anatomic/functional hypothesis would seem to favor a more balanced phonatory efficiency. The airways are respected in "welcoming" the incoming air, rather sucking it more or less violently toward the abdomen. The resonant architecture and resistances to expiratory force are also respected in the form of a comfortable compromise between the acoustic characteristics of fluent speech and the prolongation of phonatory durations. PMID- 12092505 TI - [Central and peripheral deafness among autistic and deaf children]. AB - Deafness concerns periphericals etiology but also central and psychics etiologies. Knowing the level and the mechanism is difficult with the best ORL exams. Amplification is a possibility to diagnose the reality of the lack of hearing with caution and repetition. It prepares to the use of a hearing aid which can obtain the diagnosis. PMID- 12092506 TI - [The monaural pseudo-stereophonic hearing aid (BAHA) in unilateral total deafness: a study of 29 patients]. AB - For many years, unilateral total deafness has been considered a minor handicap which does not require the fitting of a hearing aid. If the condition did give a major problem, the usual recommendation for this type of hearing loss was a CROS system which, in addition to problems of function and aesthetics, gave poor results. Faced with increasing demands from these patients, and given the good performance of bone conduction implants (BAHA), we decided to try to implant these devices on the deaf side, which allowed, by way of transcranial conduction, stimulation of the functioning ear and the re-establishment of a certain degree of stereophonic hearing. This study of 29 patients who were tested by stereophonic audiometry pre- and post-implantation shows that this type of aiding is beneficial, with improvement of the threshold of intelligibility against background noise of between 5 and 15 dB during Hirsch's test, correction of the "dead angle" on the deaf side, and improvement in localisation of sound in space. From the functional point of view, 88% of patients were satisfied with their aid, which they wore for 8 or 9 hours per day, and had a marked improvement in their hearing performance under various conditions which had proved very difficult for them before they were fitted with the aid. These results all demonstrate the effectiveness of the implant used in this way. The fact that sounds are received on the totally deaf side goes a long way towards establishing satisfactory hearing. True stereophonic hearing could not be established in any of the cases, because this requires input from two ears. It is for this reason that we have termed this type of aid as monaural pseudo-stereophonic. PMID- 12092507 TI - [Hearing evaluation on young children (less than five years old) in an under equipped center. An adaptation of the Delaroche protocol]. AB - The authors report their experience of audiometric test in childhood (less than fifty years old). It's an adaptation of Delaroche protocol which was first described by the Bordeaux's team. This prospective study has been realized in their works conditions in Abidjan (Ivory Coast) on 75 children under five years of age (average 2,64 years). This protocol is essentially based on: at first, hearing evaluation with acoustic toys; secondly, bone conduction on the vibrator; at the end, air conduction on the head set. The analysis of the results on 75 children shows that there were 35 normal examinations, 5 conductive deafness, 32 sensorineural hearing loss among which 9 partial failures (hearing threshold non evaluated), 3 complete failures (refuse the test). The authors insist on the utility of audiometric tests in early management of hearing loss in young children. PMID- 12092508 TI - Transitional care of older adults. AB - This chapter reviews 94 published research reports on transitional care of older adults by nurse researchers and researchers from other disciplines. Reports were identified through searches of MEDLINE, CINAHL, HealthSTAR, Sociological Abstracts and PsycINFO using combinations of the following search terms: transitional care, discharge planning, care coordination, case management, continuity of care, referrals, postdischarge follow-up, patient assessment, patient needs, interventions, and evaluation. Reports were included if published between 1985 and 2001, if conducted on samples age 55 and older, if relevant to nursing research, and if published in English. Intervention studies had to have a control or comparison group and a test for statistical significance. Four key findings from this review were identified. A high proportion of elders and their caregivers report substantial unmet transitional care needs, with the need for information and increased access to services consistently among the top priorities. Differences in expectations between and among patients, families, and health care providers, and the need for increased patient and family involvement in decision making, are common themes in discharge planning studies. Gaps in communication have been identified through the discharge planning process. Evidence about the effects of innovations in transitional care on quality and cost outcomes is sparse. Four main recommendations are made. Differences in older adults' transitional care needs based on race, ethnicity, and educational level, with attention to potential disparities, require further study. Studies of strategies to promote effective involvement of patients and families in decision making throughout discharge planning are needed. The development and testing of referral and other information systems designed to promote the transfer of accurate and complete information across sites of care should be a research focus. A priority for future research should be continued study of strategies to improve transitional care outcomes of older adults and their caregivers. PMID- 12092509 TI - Interventions for family members caring for an elder with dementia. AB - This chapter reviews 73 published and unpublished research reports of interventions for family members caring for an elder with dementia by nurse researchers and researchers from other disciplines. Reports were identified through searches of MEDLINE, CINAHL, Social Science Index, PsycINFO, ERIC, Social Work Abstracts, American Association of Retired Persons database, CRISP index of the National Institutes of Health, Cochrane Center database, and Dissertation Abstracts using the following search terms: caregiver, caregiving, dementia, Alzheimer's, intervention study, evaluation study, experimental, and quasi experimental design. Additional keywords were used to narrow or expand the search as necessary. All nursing research was included in the review and nonnursing research was included if published between 1991 and 2001. Studies were included if they used a design that included a treatment and control group or a one-group, pretest-posttest design (ex post facto designs were included if they used a comparison group). Key findings show that approximately 32% of the study outcomes (e.g., burden, depression, knowledge) were changed after intervention in the desired direction. In addition, several problematic issues were identified including small, diverse samples; lack of intervention specificity; diversity in the length, duration, and intensity of the intervention strategies; and problematic outcome measures. PMID- 12092510 TI - End-of-life care for older adults in ICUs. AB - This review was undertaken to present and critique the most recent (1990-2000) empirical evidence about end-of-life care for older adult patients in ICUs, their families, and care providers. The studies (including descriptive, correlational, longitudinal, and intervention) were found using a combination of these terms: (a) intensive care (units) or critical care (units), and (b) critical illness, critically ill patients, terminally ill, terminal care, life support care, or palliative care. The computerized databases searched were CINAHL and MEDLINE. Only published studies of persons 44 years of age or older, written in English, and conducted in the U.S. or Canada were included. Research was not limited to studies conducted by or written by nurses. Excluded were articles focused on physiology, for example, studies of treatment for specific conditions, and articles focused on predictors of ICU outcomes. PMID- 12092511 TI - Nursing homes and assisted living facilities as places for dying. AB - This chapter reviews the state of knowledge about nursing homes and assisted living facilities as places for dying. Reviewed are 25 published and unpublished research reports by nurse researchers and researchers from other disciplines that address the following questions: (a) What is known about how communication and shared decision-making about end-of-life care preferences occur? (b) How are symptoms assessed and managed at end-of-life? and (c) What are facility characteristics that influence end-of-life care services delivery? Reports were identified through searches of the following databases: MEDLINE, CINAHL, Health Star, PsychLit, Ageline, Ebsco, and PubMed. The following terms guided the search: advance directives, geriatric assessment or nursing, health services for the aged, hospice, residential facilities, palliative care, symptom management, and terminal care. Reports were included if published between 1990 and 2000, if relevant to nursing research on end-of-life care, if conducted on samples age 65 or older and living in nursing home or residential care settings, and if published in English. The studies reviewed were primarily descriptive. The findings indicate that little is known about end-of-life care in these settings, and that family and staff perspectives differ on the nature and quality of the services provided. Both external and internal factors influence the ability of facilities to provide end-of-life care. Recommendations are provided for further research related to nursing homes and assisted living facilities as places for dying. PMID- 12092512 TI - Home health services research. AB - This chapter reviews 69 published research reports of home health care from a health services perspective by nurse researchers and researchers from other disciplines. Reports were identified through searches of the National Library of Medicine (MEDLINE), and the Cumulative Index to Nursing and Allied Health Literature and Social Sciences Citation Index using the following search terms: home health care, health services research, and elders. Within the major areas identified, the following additional terms were specified: resource use and outcomes. Reports were included if published between 1995 and 2001, used samples age 65 and older, performed in the U.S., and published in English. Studies of all types were included. The key findings follow: (a) Most studies were a theoretical. If a theoretical model was used, it was most often the Andersen Behavioral Model. (b) Few conclusions can be drawn about resource use--increasing age and higher severity of health related problems are associated with higher numbers of home visits. The variety of measures of resource use and the study approaches (large national data sets versus single or several agency samples) limits the ability to draw conclusions on resource use. (c) There is a growing body of evidence on rehospitalization of home health care patients which indicates rehospitalization is prevalent but largely not predictable. (d) Patient outcomes research is inconclusive at this point, primarily because there are few studies that examine patient outcomes using a consistent set of measures. The main recommendations are: to study rehospitalization using a more profile-based approach to determine visit patterns that may be effective, to further specify the kinds of outcomes that may be achieved as a result of home health care and which patients might be expected to achieve positive outcomes, and to examine the integration of home health care with the broader community-based services. PMID- 12092513 TI - Telehealth interventions to improve clinical nursing of elders. AB - This chapter reviews reports of research conducted worldwide from 1966 to January 2001 on telehealth interventions in clinical nursing for elders. Reports were identified through a systematic search of MEDLINE, CINAHL, PsychInfo, ERIC, and ACM using the search terms Telemedicine or Health Information Networks, Nursing, and Research, and were restricted to those published in English. Reports of research using interactive computer technology to assess or intervene with nursing problems commonly observed in persons age 65 and older were sought. Only published reports presenting the findings of an exploratory or experimental study and exploring the association between one intervention variable and technology were included. The search resulted in 18 research reports describing eight research projects. Due to the preponderance of demonstrations and feasibility reports, the dearth of experimental investigations, and the heterogeneous nature of the few studies identified, statistical summarization was not attempted. Telehealth interventions have the potential to improve the clinical nursing care of elders because they provide alternative, equivalent approaches to assess key indicators of the physical and psychological state of elders; are acceptable to nurses, elders, and family caregivers; and may prove less costly than face-to face interventions. Telehealth approaches provide not only acceptable substitutes for discrete nursing actions but also can serve as a context within which a large range of professional gerontological nursing services can be delivered in a manner that is timely and convenient for elders. PMID- 12092514 TI - Maintaining and improving physical function in elders. AB - This chapter reviews 60 published research reports about maintaining and improving physical function in elders. Reports were identified through searches of MEDLINE and CINAHL using the following search terms: Activities of Daily Living, aged, nursing care, nurse-patient relations, geriatric nursing, nursing assessment, geriatric assessment, behavior therapy, disability evaluation, exercise, and physical. Reports were included if published in English between 1985 and 2000, if conducted in adults over age 60, and if the first author was a nurse or the reported intervention was implemented by nurses. Descriptive and experimental studies of physical function in elders were included. The results of this synthesis showed that nurse scientists have developed new instruments to measure physical function, including scales sensitive to changes in physical function caused by dementia. Nursing studies have described many of the physical and psychological factors associated with poor physical function in elders, though environmental factors and ethnic differences in physical function merit further study. Many nurse scientists are currently investigating behavioral interventions, exercise interventions, and changes in nursing care that could improve the physical function of both community-dwelling elders and those who live in residential facilities. PMID- 12092515 TI - Genetics and gerontological nursing: a need to stimulate research. AB - The purpose of this chapter is to discuss how genetics will affect gerontological nursing. The chapter will answer two questions: (1) Which aspects of genetics will be most relevant to future gerontological nursing practice? and (2) What will be the impact of genetics on the future of gerontological nursing education and research? MEDLINE was searched for relevant articles from 1995 to 2001 using the key words aging, genetics, geriatrics, nursing education, research, and gerontology. CRISP was searched using the thesaurus terms education/planning, genetics, health education, model design/development, psychological model, pubic health curriculum, behavioral/social science research, and research nursing/genetics. A total of 101 nursing and nonnursing articles were reviewed. Research reports were selected if they focused on issues related to gerontological nursing. Articles were reviewed that had application to genetic nursing, complex diseases, and genetics. The evolution of the science of genetics will revolutionize gerontological nursing and affect future nursing education and research as the concepts of genetic science and the technology they generate are translated into everyday clinical practice. Genetic discoveries in common complex diseases will affect care provided by gerontological nurses in the 21st century. Gerontological nurses must move quickly to recognize this genetic paradigm shift and to incorporate genetics issues into their nursing practice. PMID- 12092516 TI - Hearing impairment. AB - The purpose of this chapter is to review the literature on hearing impairment, specifically the impact of hearing impairment on the functioning of elders, interventions that minimize the impact of hearing loss on functioning, and identification of issues raised by the review for nursing research. Computerized (MEDLINE, PsychINFO, and CINAHL) and manual searches were used to obtain research reports from a range of disciplines. Research articles including elders (> or = 60) and published between 1989 and 2001 were included. Twenty-five articles were selected for critical review, four written by nurses. The diversity of methodologies, the criteria used to define hearing impairment, the range of sample characteristics, and the assessment measures make comparisons across studies difficult. Most studies, however, support the negative impact of hearing impairment, especially on psychosocial functioning. Measures that are condition specific are generally more effective in capturing the impact of hearing loss than generic measures. Findings related to physical disability are less consistent. The results of intervention studies suggest that hearing devices can improve psychosocial and communication outcomes, but behavioral interventions have not shown long lasting benefit. For nurses to assist elders and their families manage the impact of hearing impairment, further research is needed in several areas that have been poorly explored. These include the dyadic experience of hearing impairment, the way in which culture influences the experience of hearing loss, the needs of hearing impaired individuals across settings, the long term impact of ototoxic medications, and strategies to assist elders in coping with hearing impairment and utilizing available technologies. PMID- 12092517 TI - Pressure ulcer prevention and management. AB - This chapter reviews 218 published and unpublished research reports of pressure ulcer prevention and management by nurse researchers and researchers from other disciplines. The electronic databases MEDLINE (1966-July 2001), CINAHL (1982-June 2001), AMED (1985-July 2001), and EI Compedex Plus (1980-June 2001) were selected for the searches because of their focus on health and applied research. Moreover, evaluations of previous review articles and seminal studies that were published before 1966 are also included. Research conducted worldwide and published in English between 1930 and 2001 was included for review. Studies using descriptive, correlational, longitudinal, and randomized control trials were included. This review found that numerous gaps remain in our understanding of effective pressure ulcer prevention and management. Moreover, the majority of pressure ulcer care is derived from expert opinion rather than empirical evidence. Thus, additional research is needed to investigate pressure ulcer risk factors of ethnic minorities. Further studies are needed that examine the impact of specific preventive interventions (e.g., turning intervals based on risk stratification) and the cost-effectiveness of comprehensive prevention programs to prevent pressure ulcers. Finally, an evaluation is needed of various aspects of pressure ulcer management (e.g., use of support surfaces, use of adjunctive therapies) and healing of pressure ulcers. PMID- 12092518 TI - Elder mistreatment. AB - Elder mistreatment (EM) is a serious and prevalent syndrome that is estimated to affect between 500,000 to 1.2 million older adults in the United States annually (Pillemer & Finkelhor, 1988). This chapter reviews both the state of the published science and limitations in the knowledge base on the topic. The literature for this review was obtained through computer-assisted searches of PubMed (878 citations), the Cumulative Index of Nursing Research (CINAHL) (593 citations) and Psych-Info databases (443 citations). The search terms used were elder mistreatment, elder neglect, elder abuse, or domestic abuse of the elderly. No limit was placed on the age of publications because of the relative scarcity of research on the subject. Nonnursing articles were included because there are so few nurse researchers addressing this topic. The age limit for subjects in these studies was 65 years and older. Studies were limited to those conducted in the United States, and descriptive studies were included as they form the majority of the research to date. Findings indicate that frail, very old (over 75 years), older adults who have a diagnosis of depression or dementia are more likely to be mistreated (Dyer, Pavlik, Murphy, & Hyman, 2000; Coyne, Reichman, & Berbig, 1993; Fulmer & Gurland, 1996; Lachs & Pillemer, 1995; Lachs et al., 1997; Lachs, Williams, O'Brien, Pillemer, & Charlson, 1998; Lachs & Fulmer, 1993; Lachs, Berkman, Fulmer, & Horwitz, 1994). Those older adults who required assistance with activities of daily living had poor social networks and were at higher risk for EM (Lachs & Pillemer, 1995; Lachs et al., 1997; Lachs et al., 1998; Lachs & Fulmer, 1993; Lachs et al., 1994). Neglect, as a subcategory of EM, accounts for the majority of cases (Fulmer, Paveza, Abraham, & Fairchild, 2000; Pavlik, Hyman, Festa, & Bitondo Dyer, 2001; Fulmer & Gurland, 1996). There is still debate regarding the role of minority status, abuse in childhood, and the persons most likely to mistreat older adults. There is a critical need for replication studies and new research on this important topic. Problems with measurement, funding challenges, and the paucity of investigators conducting research on EM have left the field with several unanswered questions and some conflicting findings. This chapter summarizes the interdisciplinary literature and makes recommendations for future nursing research programs. PMID- 12092519 TI - Pain in older adults. AB - This chapter reviews 80 published research reports of pain and pain problems in older adults by nurse researchers and researchers from other disciplines. Reports were identified through searches of MEDLINE and the Cumulative Index to Nursing and Allied Health Literature (CINAHL) using the search terms pain, older adult, aged and pain, and dementia. Reports were included if published between 1985 to 2001, if conducted on samples age 60 or older, if conducted by nurses or relevant to nursing research, and if published in English. Descriptive, qualitative, correlational, longitudinal, and intervention studies were included. Key findings include the following: pain is widely prevalent in older adult populations; few studies have included minority groups; under-identification and undertreatment of pain in older adults is a consistent interpretation of research findings; pain intensity rating scales are as valid and reliable in older populations as in younger populations; current observational methods of assessing pain in cognitively impaired older adults must be used with caution; nursing intervention studies demonstrate the beneficial effects of education and interventions aimed at improved pain assessment. The main recommendations are: careful attention should be given to the conceptualization and definition of pain; examination of pain should include physiological, motivational, cognitive, and affective factors; studies evaluating undertreatment of pain should include measures of pain self-report; standardized pain measures should be used; studies of persons over the age of 85 and studies of ethnic minorities are needed; more attention should be given to nursing intervention studies and should include both pharmacological and nonpharmacological, psychosocial interventions. PMID- 12092520 TI - Interventions for persons with irreversible dementia. AB - This chapter provides an overview and critique of the theoretical and research literature by nurse researchers and researchers in other disciplines regarding interventions for persons with dementia (PWD). Reports were included if published in English between 1990 and 2000 and if a descriptive, correlational, longitudinal, or intervention design was used. Case studies and narrative descriptions were not included. No specific age criteria for study participants were applied; however, PWD are generally over age 55. The theoretical literature and various disease stages were reviewed, including clinical and behavioral indicators for disease progression. Using a variety of approaches to survey the extant literature (review of computer databases, contacts with experts in the field, ancestry method, and manual searches of key gerontology journals), over 1,200 citations were initially reviewed, allowing for approximately 375 publications undergoing thorough analysis with 157 research publications being included in this synthesis. Key findings include the identification of well supported cognitive-behavioral interventions to enhance cognitive functioning and memory, and to relieve depression in the early disease stages; multiple environmental and behavioral approaches for improvement in functioning, maintenance of activities, and alleviation of behavioral symptoms in the middle disease stages; and behavioral, interactive, and staff support and education interventions for adequate nutritional intake, urinary incontinence, and management of problematic vocalizations and other behavioral symptoms in the later disease stages. Recommendations for future studies include the need for development of operational definitions of behavioral symptoms, inclusion of the perspective of PWD, evaluation of long-term outcomes, adequate sample size, community rather than institutional-based studies, and increased intervention testing at various stages of the disease. PMID- 12092521 TI - [Early childhood infections and the development of allergy and asthma]. PMID- 12092522 TI - [Pidotimod in recurring respiratory infection in children with allergic rhinitis, asthma, or both conditions]. AB - OBJECTIVE: To demonstrate a decrease in the number of recurrent breathing infections in patients with allergic rhinitis and asthma, by means of the use of Pidotimod (a immunomodulator). MATERIAL AND METHODS: Patients of both sexes, 2 to 16 years old, that had at least four episodes of acute respiratory infection, during the previous six months, administering a dose of 400 mg of pidotimod later, twice a day. RESULTS: It was identified a correlation between the number of cases of asthma and those of acute respiratory infection of r = 0.60 with a r2 = 40. A total of 73 patients of both sexes that presented allergic rhinitis and bronchial asthma, had developed at least four acute respiratory infection episodes. Form these patients, 33 were male, a rate that corresponded to 45%, and 40 were female, that corresponded to 55% of population. The average age was 6 years old, with a range from 2 to 16 years. The number of infectious events before using Pidotimod, was 422, with an average of 5.7 for patient, and later to its use, it was 295, that corresponded to 4.04 for each patient, in 6 months. The difference among the means was significant (p < 0.005). The number of days affected by each infectious event was of 446, with an average of 6.10 for patient, and later to administration of the product it was of 308 affected days, that corresponded to an average of 4.21 days for each patient, with a statistical significant of p < 0.001. PMID- 12092523 TI - [Prevalence and risk factors for allergic conjunctivitis in children. Comparative study in two communities]. AB - OBJECTIVE: To determine the prevalence and risk factors for allergic conjunctivitis in childhood populations, in two communities with different urban and industrial development, and population density. METHODS: A questionnaire was applied to childhood from 6 to 14 years old, selected through cluster sampling. This was an epidemiologic study, observational, open, retrospective and cross sectional. This is a case-control study with cohorts. Statistical analysis consisted in frequency distributions, odds ratios, 95% confidence intervals. Statistical significance was p < 0.05. RESULTS: In the population El Higo, at Veracruz state, where sugar industry is the most important commercial activity, allergic conjunctivitis prevalence was 9.7%; allergic family antecedent was found in 79.74% of patients (p = 0.001, OR = 8.1); nicotinism at home in 12.65% (p = 0.001, OR = 0.34); factory near of patients' house in 65.82% (p = 0.001, OR = 3.5). In Tampico-Madero-Altamira suburb, the prevalence of the pathology was 10.09%; allergic family antecedent 58.89% (p = 0.001, OR = 2.52%); nicotinism at home 36.25% (p = 0.59, OR = 1.06); factory near of house 21.01% (p = 0.03, OR = 1.29). CONCLUSIONS: Results let us to know the problem that health staff will cope when facing allergic conjunctivitis; that results will be important for strategies designing, in order to establish preventive actions to take in the health-disease evolution of this pathology. PMID- 12092524 TI - [Impact of integral allergic treatment in the quality life of patients with allergic rhinitis]. AB - BACKGROUND: The quality of life is the perception of a patient about functional effects of his illness and therapy. Recently, appropriate instruments were devised to evaluate the health related with the quality of life, as the standardized and validated questionnaires for rhino-conjunctivitis. OBJECTIVE: To determine the impact of integral allergological treatment upon the quality of life of patients with allergic rhinitis that were attended as outpatients at the Service of Allergy and Clinical Immunology of Hospital Universitario de Puebla. MATERIAL AND METHODS: It was carried out a longitudinal, prospective, experimental and prolective study in patients of both sexes, older than 18 years old, with diagnosis of allergic rhinitis; life quality was evaluated by means of the application of a validated and standardized questionnaire every 15 days during 4 months. After being included in the study, patients received the integral allergological treatment conformed by educational measures, environmental control, pharmacotherapy and specific immunotherapy. We determined frequency for qualitative variables and average for quantitative variables. The averages were compared by means of variance analysis. The significance value was established down in 0.05. RESULTS: Thirteen patients were included, 61.5% of them were male. The average age was 33.2 (SD 17.0). There was a significative difference in quality of life index when patient was included in comparison with the indexes in subsequent visits. The most frequent allergic rhinitis was the moderate-severe persistent type, with a 76.9% rate. CONCLUSION: Integral allergological treatment improves quality of life of patients with allergic rhinitis. PMID- 12092525 TI - [Prevalence of latex-fruit syndrome in health workers with latex allergy]. AB - BACKGROUND: Prevalence of latex allergy in the general population is lesser than 1%. These patients have clinical and immunochemical cross-reactivity between latex and fruits; sometimes this has been referred to as the "latex-fruit syndrome" (LFS); atopy was found to be a risk factor to sensitization to fruits. OBJECTIVE: To investigate the prevalence of LFS, in a group of health care workers with latex-allergy. METHODS: Hospital employees were initially screened for latex allergy with a questionnaire; these patients were divided into the following two groups: 1) health care workers with latex-allergy, classified into two subgroups: a) with a familial history of atopy; b) without a familial history of atopy, and 2) health care workers with familial history of atopy but without latex-allergy. Skin prick tests with latex and fruits extracts (kiwi, avocado, banana and chestnut) were done. RESULTS: Based in clinical history and with confirmation by skin testing, three patients of the health care workers' group with latex allergy have LFS (prevalence of 12.5%). Sensitivity and specificity for skin prick test in health care workers with latex-fruit syndrome were: latex, kiwi and chestnut sensitivity: 100%; latex and avocado specificity: 90%; chestnut and kiwi specificity: 100%. CONCLUSION: A low prevalence of latex-fruit syndrome was detected in our population. We found a higher prevalence in females with a history of atopy. Our findings were consistent with other literature reports. PMID- 12092526 TI - [Aspirin induced asthma, urinary leukotriene E4 and zafirlukast]. AB - OBJECTIVE: To evaluate the efficiency of zafirlukast in patients with aspirin induced-asthma trough a measurement of VEF1, during the treatment and the symptoms daily reported. To evaluate the safety of treatment through a record of adversed events and laboratory and cabinet monitoring, to know the effect of zafirlukast on urine LTE4 level. MATERIAL AND METHODS: It was an open, controlled, prospective, longitudinal, observational study. Twenty patients with aspirin-induced asthma were included and 10 extrinsic asthma patients as a group control. On the aspirin-induced asthma, age range was between 28-82 years old patients with an average of 55. 17 female and three male and group control with demographic similar features. All patients were included in a 30 days wash period avoiding anti-inflammatory medication like steroids, cromons, etc., on the initial phase treatment. A determination of blood cell count, transaminases, bilirubins, immunoglobulins, GAME, electrocardiogram, thorax study and LTE4 urine analysis was carried out. Twice a day 20 mg. zafirlukast dosis was administered to both groups, VO, during eight weeks. A symptoms report diary was given to each of the patients. A weekly and beginning sphirometric was carried out along the study to determine VEF1. Same laboratory and cabinet variables were determined on the final checking. RESULTS: 17 patients from the study group were analyzed and 10 from control group, an improvement of 12% average was reported on final VEF1, in comparison to the basal for both groups (p < .001) having this relation with the clinic improvement in the diary symptoms report. LTE4 urine levels diminished in similar form on both groups. This reduction was important (p < .001). Just three limited slight cephalalgia cases were reported (7.9%). CONCLUSION: Asthmatic patients belonging to extrinsic or aspirin-induced asthma groups irrespectively showed an improvement from zafirlukast treatment on spirometric and clinic levels. No serious disadvantage was found associated to zafirlukast and side effects were slight after two months. In LTE4 level drop was found in urine, in possible relation with zafirlukast, interruption on the pulmonary chronic inflammatory process. PMID- 12092527 TI - [Hyper-IgM syndrome: mucocutaneous lesions and neutropenia]. AB - The hyper-IgM syndrome is a rare immunodeficiency disease in which the ability of B cells to switch immunoglobulin production from IgM to IgG, IgA and IgE is defective. The deficiency of IgG and IgA leads to recurrent infections of the respiratory tract, but some patients are susceptible to infection with opportunistic microorganisms, such as Cryptococcus neoformans, and also are prone to neutropenia and mucocutaneous ulcerations. We report a case of a two-year-old boy that was given the diagnosis of the hyper-IgM syndrome on the basis of low serum concentrations of IgG and IgA and high serum levels of IgM associated with C. neoformans infection, neutropenia and mucocutaneous ulcerations. Intravenous immune globulin improves dramatically the disorder, including neutropenia. To date, periodical infusion of immune globulin has prevented the development of serious infections. PMID- 12092528 TI - [Correlation between renal-uretero-bladder diseases and allergies]. AB - Some clinical reports suggest that allergic and reno-urinary diseases could be associated. Etiopathogenic aspects are described, as well as some clues of anamnesis that will help us to suspect allergic etiology. PMID- 12092559 TI - Chest X-ray quiz. Left pneumothorax and left apical drain in situ. PMID- 12092560 TI - Mutual recognition of nurse licensure. PMID- 12092561 TI - Grand Forks survives, grows from flood disaster. PMID- 12092562 TI - [Lesions of the mouth mucosa. An anamnestic and clinical study of 100 consecutive patients with mucosal lesions]. AB - The present study evaluates and analyzes anamnestic and clinical findings of 100 consecutively referred patients with oral mucosal lesions. The mean age of the cohort was 50.5 years (range 8-91 years) with a female rate of 60 percent. A total of 9 malignant neoplasia were found among the 100 mucosal lesions, including 7 squamous cell carcinomas and 2 mucoepidermoid carcinomas. In addition, 11 cases presented with leukoplakia and 4 cases with lichen planus, both recognized as possible precancerous conditions. These figures support the essential role of the dentist with respect to initial diagnosis and prevention of oral cancer. In addition, stomatologic follow-up examinations are recommended to be performed by each and every general practitioner. In the near future, the dentist will assess oral mucosal lesions more frequently due to the increasing number of elderly patients and to the fact that elderly people present with more lesions compared to younger patients. consistent to literature data, patients with leukoplasia and squamous cell carcinoma had the highest rates of tobacco and alcohol consumption. Therefore, the dentist may also become active in giving preventive information to the patient. the results of the present study showed, that--by employing a standardized examination--a correct clinical diagnosis was identical to the final diagnosis established by laboratory techniques. PMID- 12092563 TI - [Patient satisfaction as outcome measure in clinical studies of oral health]. AB - Patient based outcomes are increasingly used to supplement clinical parameters in clinical studies. Patient satisfaction is one of the most important of these outcomes. Patient satisfaction is not equivalent with quality of life. Patient satisfaction can be measured with a questionnaire approach, using items tapping specific aspects--chewing function or esthetics--of dental treatment. Questionnaire results are scored using either Likert or visual analog scales. Clinical studies confirm that patient satisfaction performs well as a patient based outcome for the evaluation of dental treatments. Significant differences exist in patient satisfaction with different implant prostheses. Removable prostheses are perceived as easier to clean while fixed ones are rated more stable. these factors have a significant influence on the overall satisfaction with treatment and on the choice of treatment. Results from studies using patient based outcomes can generate improved information for patients, providers and insurers concerning treatment alternatives, and help to avoid treatment failures. PMID- 12092564 TI - [Facts and statistics from the study of dentists in 2001]. PMID- 12092565 TI - NGNA makes recommendations to the National Quality Forum. PMID- 12092566 TI - Is Cr(VI) toxicity to Daphnia magna modified by food availability or algal exudates? The hypothesis of a specific chromium/algae/exudates interaction. AB - The long-term effects of Cr(VI) on life history traits of Daphnia magna Straus were studied in relation to different feeding conditions or to the presence of Scenedesmus acutus exudates, which had proved to reduce chromium toxicity to the alga. In high feeding conditions the concentration of 14 microg Cr(VI)/L had negative effects on survival, growth and fecundity. In low feeding conditions the effects on growth were not evident, but daphnids changed the reproductive strategy normally adopted in case of food shortage and showed a daily rate of newborn production higher than control; however, newborn were of a smaller size and less resistant to starvation. The most relevant effect of Cr was a shortening of the daphnid life span, which was independent of the feeding conditions and occurred also when growth and daily rate of neonate production were not affected. The algal exudates resulted ineffective in reducing Cr toxicity to daphnids, but caused a strong decrease in daphnid fecundity. Further experiments on S. acutus demonstrated that the detoxifying effect observed in the algae might involve a specific Cr/algae/exudates interaction, which occurs only when the algae are previously subjected to a short-time stress by chromium. PMID- 12092567 TI - P removal from anaerobic supernatants by struvite crystallization: long term validation and process modelling. AB - In this work, a model for phosphorus crystallization in a fluidized bed reactor, able to describe the experimental results obtained during a semiscale pilot plant, is presented. In particular, the validity of the model proposed has been evenly extended with respect to a previous experiment, even at a lower contact time, and the length of each experiment has been increased, in order to verify the behaviour of the process for long term applications and to evaluate the maximum crystal growth of the system. Moreover, the state of the art of the available processes for phosphorus removal from wastewater is presented, together with a detailed review of the several models so far developed to describe the phosphorus crystallization mechanisms. PMID- 12092568 TI - The leaching of inorganic species from activated carbons produced from waste tyre rubber. AB - Waste tyre rubber can be used as a precursor for the production of high quality activated carbons. However, there is concern that inorganic impurities present in the rubber feed may restrict their use in liquid phase applications with high purity requirements. This paper presents an investigation of the presence and the leaching of inorganic species from activated carbons derived from waste tyre rubber. For the purpose of this work, a number of carbons were produced, characterised for their BET surface area and analysed for their inorganic composition. Subsequently, a number of tests were performed to evaluate the leaching of different inorganic species into solution at various pH values and carbon doses. Results showed that rubber-derived carbons contained elevated concentrations of sulphur and zinc, as well as traces of other metals such as lead, cadmium, chromium and molybdenum. Inorganic levels were significantly affected by production conditions, particularly degree of carbon activation and the nature of the gasification agent. However, leaching tests showed that the availability of these species in neutral pH conditions was very limited. Results demonstrated that, when using carbons doses comparable to those employed in water treatment works, only sulphur levels exceeded, in some occasions, health based quality standards proposed for drinking water. PMID- 12092569 TI - Application of catalytic wet air oxidation for the treatment of H-acid manufacturing process wastewater. AB - Four types of powder catalysts whose main active components are copper (Cu), cerium (Ce), cadmium (Cd) and cobalt-bismuthide (Co-Bi) are prepared with the method of the co-deposition and are evaluated through the catalytic wet air oxidation (CWAO) treatment of H-acid solution. The comparison of the efficiencies of different catalysts shows that Ce3Cu1 (3:1) catalyst is the best one. When the reaction temperature is 200 degrees C, oxygen partial pressure is 3 MPa, pH value is 12, and reaction time is 30 min, the COD removal rate is over 90%. All the H acid is decomposed in 5 min and is oxidized into NH4+, SO4(2-) formic acid, acetic acid and other end products. The pH value can greatly affect the COD removal and the production of organic acid. CWAO process not only can get a high reaction rate, but also can oxidize the short-chain organic acid. PMID- 12092570 TI - The effects of tertiary wastewater treatment on the prevalence of antimicrobial resistant bacteria. AB - The effects of tertiary wastewater treatment on the prevalence of antimicrobial resistant bacteria were investigated in two large-scale municipal treatment plants during a period of six months. Total and relative numbers of resistant bacteria were determined in raw sewage, treated sewage and anaerobically digested sludge by bacteriological counts on media selective for coliforms (MacConkey agar) and Acinetobacter spp. (Baumann agar). In addition, the level of antimicrobial susceptibility was determined by the disc-diffusion method in 442 Acinetobacter isolates identified by colony hybridisation with a genus-specific DNA probe. Independent of the different antibiotics and media used, the total numbers of resistant bacteria in treated sewage were 10-1000 times lower than in raw sewage. Based on linear regression analysis of data on bacteriological counts, the prevalences of antimicrobial-resistant presumptive coliforms and Acinetobacter spp. in treated sewage and digested sludge were not significantly higher compared with raw sewage. On the contrary at one plant, statistically significant decreases were observed in the prevalence of ampicillin-resistant presumptive Acinetobacter spp. (p = 0.0188) following sewage treatment, and in the prevalence of either ampicillin-resistant presumptive Acinetobacter spp. (p = 0.0013) or ampicillin- and gentamicin-resistant presumptive coliforms (p = 0.0273 and p = 0.0186) following sludge treatment. The results obtained by bacteriological counts were confirmed by antimicrobial susceptibility testing of Acinetobacter isolates. Based on logistic regression analysis, isolates from treated sewage and digested sludge were generally not significantly more resistant compared with isolates from raw sewage. Based on these evidences, it was concluded that tertiary wastewater treatment did not result in a selection of antimicrobial resistant bacteria. PMID- 12092571 TI - Faecal contamination of source-separated human urine based on the content of faecal sterols. AB - Transmissible pathogens in source-separated human urine, intended for reuse in agriculture, mainly originate from faeces that cross-contaminate the urine. The health risks associated with the enteric pathogens will largely be dependent on their initial concentration and their inactivation during storage in the urine. Faecal sterols have proven stable in urine and can, rather than indicator bacteria, be used to quantify the faecal cross-contamination. In this study, urine collection tanks were sampled and ratios between various faecal sterols were used to determine if the urine was contaminated by faeces. Twenty-two percent of samples from the upper part of the tanks and 37% of samples from the bottom sludge were found to be contaminated. Coprostanol concentrations in the contaminated urine samples corresponded to a mean faecal contamination of 9.1+/ 5.6 mg l(-1) urine. E. coli was absent in a majority of the samples. Faecal streptococci and clostridia were enumerated but not found to correlate with coprostanol concentrations in contaminated samples. PMID- 12092572 TI - Degradation of TCE, Cr(VI), sulfate, and nitrate mixtures by granular iron in flow-through columns under different microbial conditions. AB - Flow-through aquifer columns packed with a middle layer of granular iron (Fe0) were used to study the applicability and limitations of bio-enhanced Fe0 barriers for the treatment of contaminant mixtures in groundwater. Concentration profiles along the columns showed extensive degradation of hexavalent chromium Cr(VI), nitrate, sulfate, and trichloroethene (TCE), mainly in the Fe0 layer. One column was bioaugmented with Shevanella algae BRY, an iron-reducing bacterium that could enhance Fe0 reactivity by reductive dissolution of passivating iron oxides. This strain did not enhance Cr(VI), which was rapidly reduced by iron, leaving little room for improvement by microbial participation. Nevertheless, BRY-enhanced nitrate removal (from 15% to 80%), partly because this strain has a wide range of electron acceptors, including nitrate. Sulfate was removed (55%) only in a column that was bioaugmented with a mixed culture containing sulfate-reducing bacteria. Apparently, these bacteria used H2 (produced by Fe0 corrosion) as electron donor to respire sulfate. Most of the TCE was degraded in the zone containing Fe0 (50 70%), and bioaugmentation with BRY slightly increased the removal efficiency to about 80%. Microbial colonization of the Fe0 surface was confirmed by scanning electron microscopy. PMID- 12092573 TI - Non-steady state modeling of extracellular polymeric substances, soluble microbial products, and active and inert biomass. AB - We present a modeling approach that quantifies the unified theory presented in the companion paper. In this approach, we use mathematical modeling to quantify the relationships among three solid species--bacteria, extracellular polymeric substances (EPS), and inert residual biomass-two soluble microbial products (SMP), original substrate, and an electron acceptor. According to the model, donor electrons are used for the synthesis of biomass, EPS, and utilization associated products. Residual inert biomass and biomass-associated products are produced from the decay of active biomass and the hydrolysis of EPS, respectively. The model includes mass balance equations that consistently describe the flow of electrons among the components. It is solved with a set of parameters appropriate to the experimental study of Hsieh et al. (Biotech. Bioeng. 44 (1994) 219). Model outputs capture all trends observed in steady-state CSTR experiments and transient batch experiments. This agreement supports that the unified theory correctly captures the interconnections among SMP, EPS, and active and inert biomass. PMID- 12092574 TI - Anaerobic/aerobic treatment of coloured textile effluents using sequencing batch reactors. AB - Conventional biological wastewater treatment plants do not easily degrade the dyes and polyvinyl alcohols (PVOH) in textile effluents. Results are reported on the possible advantages of anaerobic/aerobic cometabolism in sequenced redox reactors. A six phase anaerobic/aerobic sequencing laboratory scale batch reactor was developed to treat a synthetic textile effluent. The wastewater included PVOH from desizing and an azo dye (Remazol Black). The reactor removed 66% of the applied total organic carbon (load F: M 0.15) compared to 76% from a control reactor without dye. Colour removal was 94% but dye metabolites caused reactor instability. Aromatic amines from the anaerobic breakdown of the azo dyes were not completely mineralised by the aerobic phase. Breakdown of PVOH by the reactor (20-30%) was not as good as previous reports with entirely aerobic cultures. The anaerobic cultures were able to tolerate the oxygen and methane continued to be produced but there was a deterioration in settlement. PMID- 12092575 TI - A flow-through chamber for toxicity testing of 80-200 microm organisms. AB - A flow-through chamber was developed for toxicity testing of mobile species that require microscopic observation to assess viability and development; no sub sampling was required in this method. Organisms were held in 1.5 mL volumes in 30 mm plastic petri plates adapted with two side sections containing 35 microm screening to provide drainage. Test solutions continuously entered via a drip tube; five 1.5 mL volumes were exchanged per hour. The 30 mm organism dish was housed in an outer dish (60 mm petri plate) that captured effluent for water quality measurements. Direct observation of organisms in the thin water layer occurred without refocusing the microscope. The flow-through chamber was used to test the toxicity of copper to Mercenaria mercenaria (hard clam) as they developed from free swimming larvae (approximately 80 + microm) through metamorphosis to the settlement stage (approximately 200 microm). The flow through chamber performed well and good water quality was maintained throughout the assay. Metamorphosis occurred at the appropriate developmental time with relatively low variability between replicates. Copper, at a concentration of 14 microg/L Cu, was shown to significantly increase mortality and inhibit metamorphosis after 36 h of exposure. PMID- 12092576 TI - Sorption of benzo[a]pyrene to sediment contaminated by acid mine drainage: contrasting particle concentration-dependencies in river water and seawater. AB - The sorption of benzo[a]pyrene (BaP) to sediment contaminated by acid mine drainage from the River Carnon, SW England, has been studied as a function of particle concentration in river water and seawater. The sediment is iron-rich and has a high specific surface area, and previous studies have demonstrated that it exhibits positive surface charge when suspended in native river water (pH approximately 5), and, through an increase in pH (to about 8) and the adsorption of organic matter, negative charge when suspended in seawater. Unit sorption of BaP exhibited an inverse dependency on particle concentration (particle concentration effect. PCE) in seawater, but was relatively invariant in river water. Consequently, BaP was apparently salted out at particle concentrations below about 100 mg l(-1), and salted in at higher particle concentrations. The absence of a PCE (and inherent salting in) for such a hydrophobic compound is an unusual, and possibly unique observation, and is inconsistent with a third (e.g. colloidal) phase model. More likely, a PCE in seawater is caused, in part, by salt induced particle-particle or particle-organic matter interactions which result in increasing particle aggregation or sorption irreversibility with increasing particle concentration. Although the results have limited environmental application and do not conclusively resolve any causal mechanisms for the PCE, they suggest that, with respect to neutral organic compounds, adsorbed organic matter is a pre-requisite for a PCE in the natural environment, and the effect should not solely or generally be attributed to the existence of a third phase. PMID- 12092577 TI - Toxic effects of Irgarol 1051 on phytoplankton and macrophytes in Lake Geneva. AB - Irgarol 1051 is a recent herbicidal compound, inhibitor of photosynthesis, used in antifouling paints. This toxic is persistent in aquatic environments, with low abiotic and biotic degradation, highly phytotoxic, and has already been detected in estuaries and coastal areas, with suspected negative impacts on non-target organisms (aquatic plants and algae). We measured the toxicity of Irgarol 1051 to macrophytes and phytoplankton from Lake Geneva (between Switzerland and France) by determining chlorophyll fluorescence yield, and phytoplankton primary production. Long-term toxicity for phytoplankton was estimated in a microcosm study, and growth inhibition tests were performed with isolated algal strains. The concentration of Irgarol 1051 was analysed in the water, and the most polluted site showed a higher level (up to 135 ng/L) than the lowest observed effect concentration for phytoplankton (8-80 ng/L), while the macrophytes appeared to be more tolerant to Irgarol 1051 in short-term tests. The microcosm study showed that phytoplankton structure might be even more sensitive to Irgarol 1051. PMID- 12092578 TI - Three-dimensional pollutant transport model for the Pearl River Estuary. AB - In this paper, the development and implementation of a three-dimensional, numerical pollutant transport model, which is based on an orthogonal curvilinear coordinate system in the horizontal direction and a sigma coordinate system in the vertical direction, is delineated. An efficient as well as simple open boundary condition is employed for pollutant transport in this mathematical model. It is then applied to model the distribution and transport of Chemical Oxygen Demand in the Pearl River Estuary. The results from the numerical simulations illustrate that the transboundary or inter-boundary effects of pollutants, between the Guangdong Province and the Hong Kong Special Administrative Region due to the wastewater discharged from the Pearl River Delta Region, are quite strong. PMID- 12092579 TI - Applying a novel autohydrogenotrophic hollow-fiber membrane biofilm reactor for denitrification of drinking water. AB - We conducted a series of pseudo-steady-state experiments on a novel hollow-fiber membrane biofilm reactor used for denitrification of oligotrophic waters, such as drinking water. We applied a range of nitrate loadings and hydrogen pressures to establish under what conditions the system could attain three goodness-of performance criteria: partial nitrate removal, minimization of hydrogen wasting, and low nitrite accumulation. The hollow-fiber membrane biofilm reactor could meet drinking-water standards for nitrate and nitrite while minimizing the amount of hydrogen wasted in the effluent when it was operated under hydrogen-limited conditions. For example, the system could achieve partial nitrate removals between 39% and 92%, effluent nitrate between 0.4 and 9.1 mg N/l, effluent nitrite less than 1 mg N/l, and effluent hydrogen below 0.1 mg H2/l. High fluxes of nitrate and hydrogen made it possible to have a short liquid retention time (42 min), compared with 1-13 h in other studies with hydrogen used as the electron donor for denitrification. The fluxes and concentrations for hydrogen, nitrate, and nitrite obtained in this study can be used as practical guidelines for system design. PMID- 12092580 TI - Combined biological and ozone treatment of log yard run-off. AB - Batch biological treatment of log yard run-off reduced biochemical oxygen demand (BOD), chemical oxygen demand (COD) and tannin and lignin (TL) concentration by 99%, 80%, and 90%, respectively. Acute (Microtox) toxicity was decreased over treatment, from an initial EC50 of 1.83% to a value of 50.4% after 48 h of treatment. Kinetics of biodegradation were determined using respirometry and fitted using the Monod and Tessier model. For the Monod model the maximum substrate uptake rate, and Ks values determined were 0.0038 mg BOD/mgVSS min, and 1.4 mg/L, respectively. The efficacy of ozone as a pre- and post- biological treatment stage was also assessed. During ozone pretreatment, TL concentration and acute toxicity were rapidly reduced by 70% and 71%, respectively. Pre ozonation reduced BOD and COD concentration by < 10%, however a larger fraction of residual COD was non biodegradable after ozonation. Biologically treated effluent was subjected to ozonation to determine whether further improvements in effluent quality could be achieved. A reduction in COD and TL concentration was observed during ozonation, however no further improvement in toxicity was observed. Ozonation increased BOD by 38%, due to conversion of COD to BOD. PMID- 12092581 TI - Simultaneous uptake of anionic surfactants and micellar-solubilized contaminants using anion-exchange resins. AB - This research studied simultaneous uptake of anionic surfactants and micellar solubilized organic contaminants by anion-exchange resins. Anionic surfactant molecules adsorbed onto the positively charged resin mainly through electrostatic attraction, while the micellar-solubilized contaminants were excluded from aqueous solutions once the remaining micelles could no longer solubilize them. Data suggest that the excess contaminants adsorbed onto the resin skeleton and admicelle layer formed on the resin surface through hydrophobic interactions and eventually partitioned into the resin gel phase matrix. In batch adsorption, the contaminant solubilization capacity did not decrease linearly with respect to surfactant concentration decrease due to the increased solution counterion activity during anion exchange, and caused "delayed" contaminant uptake relative to that of the surfactant. No such effect occurred in continuous column adsorption, where the surfactant and contaminant breakthrough occurred simultaneously. Surfactant head and tail group properties, along with resin structure and particle size significantly affected surfactant and contaminant uptake rates. Relative to recovering the surfactant, the high exchange potential of the anionic surfactant prevented effective surfactant desorption, even at high electrolyte concentration and in the presence of a cosolvent. The resin matrix also had high affinity for the partitioned contaminant, and the contaminant elution from the resin seemed to be controlled by equilibrium partitioning. PMID- 12092582 TI - Utilization of a metabolic uncoupler, 3,3',4',5-tetrachlorosalicylanilide (TCS) to reduce sludge growth in activated sludge culture. AB - This paper studies the feasibility of using 3,3',4',5-tetrachlorosalicylanilide (TCS) as a metabolic uncoupler to reduce the sludge growth in activated sludge cultures. The results have confirmed that TCS is an effective chemical agent in limiting the sludge growth when its concentration is >0.4 mg/L. It was demonstrated that TCS was able to reduce sludge growth rate by around 40% when the TCS concentration was 0.8mg/L. It was also revealed that substrate removal capability was not affected adversely by the presence of TCS when TCS was continuously dosed in a range of 0.5-1.0 mg/L during the 30-day operation of activated sludge batch cultures. Such a sludge growth reduction is associated with the enhancement of microbial activities and an increase in the percentage of active bacteria over the total microbial population. In the 30-day operation of the cultures, the TCS dosing at a 1-mg/L level did not undermine the treatment performance in terms of the substrate removal efficiency. This work has demonstrated that it might be feasible to apply TCS in activated sludge systems to limit the excess sludge production. PMID- 12092583 TI - Temporal and spatial variations in kinetics of alkaline phosphatase in sediments of a shallow Chinese eutrophic lake (Lake Donghu). AB - Monthly sediment and interstitial water samples were collected in a shallow Chinese freshwater lake (Lake Donghu) from three areas to determine if alkaline phosphatase activity (APA) plays an important role, in phosphorus cycling in sediment. The seasonal variability in the kinetics of APA and other relevant parameters were investigated from 1995-1996. The phosphatase hydrolyzable phosphorus (PHP) fluctuated seasonally in interstitial water, peaking in the spring. A synchronous pattern was observed in chlorophyll a contents in surface water in general. The orthophosphate (o-P) concentrations in the interstitial water increased during the spring. An expected negative relationship between PHP and Vmax of APA is not evident in interstitial water. The most striking feature of the two variables is their co-occurring, which can be explained in terms of an induction mechanism. It is argued that phosphatase activity mainly contributes to the driving force of o-P regeneration from PHP in interstitial water, supporting the development of phytoplankton biomass in spring. The Vmax values in sediment increased during the summer, in conjunction with lower Km values in interstitial water that suggest a higher affinity for the substrate. The accumulation of organic matter in the sediment could be traced back to the breakdown of the algal spring bloom, which may stimulate APA with higher kinetic efficiency, by a combination of the higher Vmax in sediments plus lower Km values in interstitial water, in summer. In summary, a focus on phosphatase and its substrate in annual scale may provide a useful framework for the development of novel P cycling, possible explanations for the absence of a clear relationship between PHP and APA were PHP released from the sediment which induced APA, and the presence of kinetically higher APA both in sediment and interstitial water which permitted summer mineralization of organic matter derived from the spring bloom to occur. The study highlighted the need for distinguishing functionally distinct extracellular enzymes between the sediment and interstitial water of lakes. PMID- 12092584 TI - Electron spin resonance study of copper biosorption by bacteria. AB - The biosorption of copper by bacteria was studied by using ESR spectroscopy. Among bacteria tested, Arthrobacter nicotianae has the most excellent ability to sorb copper. The biosorption of copper by Arthrobacter cells was so rapid, affected by the solution pH, and obeys the Langmuir adsorption equilibrium. The electron spin resonance (ESR) spectra of Cu(II) in bacterial cells are axial type, having a major absorption to higher field at gperpendicular and lesser absorption to lower field at gparallel with four lines. The ESR parameters showed that Cu(II) in the cells has the tetragonally distorted octahedral structure with nitrogen and oxygen as ligand atoms, which suggests that most of copper in bacterial cells combined with amino acid residues in the cell surface proteins. The variation of spectral patterns among bacteria could explain as the change of ligand circumstances caused by the pH of the cell surface. PMID- 12092585 TI - The effect of UV light on the inactivation of Giardia lamblia and Giardia muris cysts as determined by animal infectivity assay (P-2951-01). AB - This study measured the effect of germicidal ultraviolet (UV) light on Giardia lamblia and Giardia muris cysts, as determined by their infectivity in Mongolian gerbils and CD-1 mice, respectively. Reduction of cyst infectivity due to UV exposure was quantified by applying most probable number techniques. Controlled bench-scale, collimated-beam tests exposed cysts suspended in filtered natural water to light from a low-pressure UV lamp. Both G. lamblia and G. muris cysts showed similar sensitivity to UV light. At 3 mJ/cm2, a dose 10-fold lower than what large-scale UV reactors may be designed to provide, > 2-log10 (99 percent) inactivation was observed. These results, combined with previously published data showing other protozoa and bacteria have similar, high sensitivity to UV light, establish that UV disinfection of drinking water is controlled by viruses which may require over 10-fold more UV dose for the same level of control. PMID- 12092586 TI - Improving the nitrogen removal efficiency of an A2/O based WWTP by using an on line knowledge based expert system. AB - The results obtained using an expert system to control an activated sludge process involving nutrient removal are reported. The study was conducted at a pilot plant using an Anaerobic/Anoxic/Oxic (A2/O) scheme for which a distributed control system was specially developed. The system allows various expert operational approaches to be developed with a view to minimize nitrogen levels in the outlet while using the minimum amount of energy. The proposed distributed control system is supervised by a Knowledge Based Expert System (KBES) constructed with G2 (a tool for expert system development) and permits the on line implementation of every operating strategy of the experimental system. A set of experiments involving variable loads and flow-rates was carried out. It revealed that the amount of removed nitrogen could be increased by 11% compared to the usual operating conditions. This increase resulted in a decrease in the amount of total nitrogen and ammonium nitrogen in the outlet by 49% and 64%, respectively. These improvements were achieved with little energy cost because the performance of the treatment plant was optimised using operating rules implemented in real time. PMID- 12092587 TI - Kinetics of the aerobic biological degradation of shredded municipal solid waste in liquid phase. AB - The organic fraction of municipal solid waste (OFMSW) should be utilised by means of biological methods. The biodegradation of solid wastes can be intensified owing to application of the bioreactors. Estimation of the optimum values of the organic load is one of the most important tasks for the aerobic biodegradation processes. The kinetic model of biological oxidation of the organic wastes has been presented in this paper. The experiments were carried out in batch 6-l working volume stirred tank bioreactors at constant temperature of 25 degrees C. Initial total solids have been at the levels of 15, 19, 34, 55 and 66 g l(-1). The kinetics of microbial decomposition of organic substances was described by means of an unstructured model. The satisfactory time courses for substrate chemical oxygen demand in the solid (CODs) and liquid phase (CODL) and biomass concentration (RNA) have been achieved. Also, the influence of the initial TS on the kinetics of the biodegradation process was investigated and the optimum value of initial TS for this type of processes was estimated at 34-55 g l(-1). PMID- 12092588 TI - Toxicity and microcystin content of extracts from a Planktothrix bloom and two laboratory strains. AB - Toxic effects and microcystin content from various extracts of a Planktothrix agardhii bloom and two different strains of Planktothrix agardhii, HUB 076, and NIVA 34 were investigated. Extracts were obtained with solvents of different polarity such as hexane, dichloromethane, methanol, and water. Additionally, different pre-treatments were used to break the cells before extraction. Acute toxicity was determined with the fairy shrimp Thamnocephalus platyurus, subchronic effects were detected in embryos and larvae of the zebrafish Danio rerio. The extracts affected the test species to a different extent. Effects were severe in polar extracts (water and methanol) of all strains tested. Although the strain NIVA 34 did not contain any microcystins, aqueous extracts of this strain showed the highest acute toxicity to the crustacean species tested (LC50= 0.08 mg ml(-1)). In contrast aqueous extracts of the Planktothrix bloom containing high amounts of microcystins were less acutely toxic (LC50 = 0.46 mg ml(-1)). Our results indicate the existence of further toxic metabolites in different Plankorothrix agardhii strains. PMID- 12092589 TI - Visualization of oxygen transfer across the air-water interface using a fluorescence oxygen visualization method. AB - Oxygen concentration fields in a water body were visualized by the fluorescence oxygen visualization (FOV) method. Pyrenebutyric acid (PBA) was used as a fluorescent indicator of oxygen, and an intensive charge coupled-device (ICCD) camera as an image detector. Sequential images (over 2000 images) of the oxygen concentration field around the surface water of the tank (1 x 1 x 0.75 m3) were produced during the 3 h experiment. From image processing, the accurate pathway of oxygen-rich, cold water at the water surface was also visualized. The amount of oxygen transferred through the air-water interface during the experiment was measured and the oxygen transfer coefficient (K(L)) was determined as 0.22 m/d, which was much higher than that is expected in molecular diffusion. Results suggest that vertical penetration of cold water was the main pathway of oxygen in the water body in the tank. The average velocity of cold water penetrating downward in water body was also measured from consecutive images and the value was 0.3-0.6 mm/s. The FOV method used in this research should have wide application in experimental fluid mechanics and can also provide a phenomenological description of oxygen transfer under physically realizable natural conditions in lakes and reservoirs. PMID- 12092590 TI - The effect of CO2 concentration on a nitrifying chalk reactor. AB - The effect of CO2 concentration on nitrification rate was studied in a fluidized bed reactor using chalk (solid calcium carbonate) as the biomass carrier and buffering agent. Using one chalk type and uniform particle size, carbon dioxide was found to limit the nitrification rate in the reactor at concentrations up to 0.3 mmol l(-1). At this concentration the nitrification rate was about 2.5-2.7g NH4+-Nl reactor(-1) d(-1). The pH established in the reactor varied between 4.5 and 5.5, remarkably with lower pH obtained remarked at higher nitrification rates. Kinetic parameters for nitrification rate with CO2 as the rate limiting substrate were determined: a Michaelis-Menten constant, Km, of 0.013 mmol l(-1) CO2 and a maximum ammonium oxidation rate of 2.33g NH4+-Nl reactor(-1) d(-1). PMID- 12092591 TI - Nutrient budget for Saguling Reservoir, West Java, Indonesia. AB - A preliminary nutrient budget for Saguling Reservoir is reported as a first attempt to quantify the behaviour of nutrients entering this reservoir. This work is part of a larger Indonesia-Australia collaborative research and training project, involving Padjadjaran University and Monash University, established to study nutrient dynamics in Saguling Reservoir. Saguling Reservoir, the first of a chain of three large reservoirs (Saguling, Cirata and Jatilahur), built on the Citarum River in central Java, was completed in 1985. It has already become highly polluted, particularly with domestic and industrial effluent (organic matter, nutrients, heavy metals) from the urban areas of Bandung (population 2 million). The reservoir experiences major water quality problems, including excessive growths of floating plants, toxic cyanobacterial blooms and regular fish-kills. The work reported in this paper shows that Saguling receives a very large nutrient load from the city of Bandung and because of this, is highly eutrophic. It is unlikely that the water quality of Saguling will improve until a substantial part of Bandung is sewered and adequate discharge controls are placed on the many industries in the region upstream of the reservoir. PMID- 12092592 TI - Fish test for endocrine-disruption and estimation of water quality of Japanese rivers. AB - The LC50 values (72 h) of 17beta-estradiol (E2), p-nonylphenol (NP) and bis phenol-A (BPA) to adult male and female medaka were 3.5 and 3.5, 0.85 and 0.87, and 6.8 and 8.3 mg L(-1), respectively; the LC50 values to embryos were 0.46, 0.13 and 5.1 mg L(-1), respectively. The IC50 values for inhibition to egg hatching were 0.47, 0.85 and 9.0 mg L(-1), respectively. These values were much higher than concentrations detected in river water in Japan and the chemicals were considered to have no lethal effect on the fish in an aquatic environment. Mature male medaka was continuously exposed to 0.005, 0.05 or 1.0 microg L(-1) of E2, or to 0.1, 10 or 100 microg L(-1) of NP or BPA. Female specific proteins (FSP) were induced in the blood of male medaka that were exposed for 5 weeks to E2 higher than 0.005 microg L(-1), NP higher than 0.1 microg L(-1), or BPA higher than 10 microg L(-1). Based on these FSP inducible concentrations and reported concentrations of E2, NP and BPA in Japanese river water, some river water contaminated by E2 or NP could be estimated as the FSP inducible in male medaka. PMID- 12092593 TI - Immunohistochemical comparison of uterine papillary serous and papillary endometrioid carcinoma: clues to pathogenesis. AB - Twenty-four predominantly papillary carcinomas of the endometrium, 10 serous and 14 endometrioid, were compared using a variety of immunohistochemical antibodies, including p53, estrogen and progesterone receptors, carcinoembryonic antigen, and E-cadherin. These were selected to attempt to find clues to explain the disparate behavior of these two tumor subtypes. We found that 6 of 8 (75%) serous carcinomas had a p53 reactivity score of 300, whereas 90% of endometrioid tumors had a p53 reactivity score of less than 20 (p = 0.0008). Combined estrogen and progesterone hormone reactivity was positive in 13 (100%) of endometrioid lesions compared with 4 of 8 (50%) of serous lesions (p = 0.0117). The significantly greater p53 expression and its significantly diminished hormone receptor expression indicate that papillary serous carcinomas belong to the type II group of endometrial carcinomas that occur in a background of atrophic endometrium, are high grade, present with high stage disease, and have a poor prognosis. In contrast, papillary endometrioid carcinomas, which belong to type I carcinomas, often arise in a background of estrogen-stimulated endometrial hyperplasia, are usually well-differentiated, and have a good prognosis. Early p53 mutations in papillary serous carcinoma as well as in endometrial intraepithelial serous carcinoma may partially explain their proclivity for early intra-abdominal dissemination. Carcinoembryonic antigen expression was similar in both groups and therefore is not useful to characterize possible differences in the cell of origin. The reactivity scores for E-cadherin were also similar in the two tumor subtypes, thus not supporting the hypothesis that decreased cell to cell adhesion molecules might contribute to early dissemination of serous lesions. PMID- 12092594 TI - United Kingdom divided as Scotland introduces free personal care for elderly people. PMID- 12092595 TI - Netherlands: long term care paid by compulsory insurance. PMID- 12092596 TI - Germany: everyone has to be insured for cost of long term care. PMID- 12092597 TI - Spain: old people frequently live with their families. PMID- 12092598 TI - Ireland: state pays maximum of 124 pounds sterling a week to the cost of a nursing home bed. PMID- 12092599 TI - Australia: federal government subsidises long term care by up to 22,000 pounds sterling a year. PMID- 12092600 TI - US: Medicaid pays for half of nursing home bills, but better off do not qualify. PMID- 12092602 TI - Harm from smoking is even greater than previously thought. PMID- 12092604 TI - Eating meat more than 10 times a week almost doubles chances of bowel cancer. PMID- 12092605 TI - Alteplase for stroke. Patients and doctors are being misled by promotional pressures. PMID- 12092606 TI - Alteplase for stroke. American Heart Association explains how guidelines were formulated. PMID- 12092607 TI - Alteplase for stroke. Financial information is needed to ensure objectivity. PMID- 12092608 TI - Alteplase for stroke. Why were these authors of the commentaries chosen? PMID- 12092609 TI - PFI is here to stay. Select committee's report should have been given greater attention. PMID- 12092613 TI - I have been bullied because of my illness. PMID- 12092615 TI - Crisis of confidence: Britain's body parts scandal still haunts the health service. PMID- 12092614 TI - Oxygen control of ethylene biosynthesis during seed development in Arabidopsis thaliana (L.) Heynh. AB - An unforeseen side-effect on plant growth in reduced oxygen is the loss of seed production at concentrations around 25% atmospheric (50 mmol mol-1 O2). In this study, the model plant Arabidopsis thaliana (L.) Heynh. cv. 'Columbia' was used to investigate the effect of low oxygen on ethylene biosynthesis during seed development. Plants were grown in a range of oxygen concentrations (210 [equal to ambient], 160, 100, 50 and 25 mmol mol-1) with 0.35 mmol mol-1 CO2 in N2. Ethylene in full-sized siliques was sampled using gas chromatography, and viable seed production was determined at maturity. Molecular analysis of ethylene biosynthesis was accomplished using cDNAs encoding 1-aminocyclopropane-1 carboxylic acid (ACC) synthase and ACC oxidase in ribonuclease protection assays and in situ hybridizations. No ethylene was detected in siliques from plants grown at 50 and 25 mmol mol-1 O2. At the same time, silique ACC oxidase mRNA increased three-fold comparing plants grown under the lowest oxygen with ambient controls, whereas ACC synthase mRNA was unaffected. As O2 decreased, tissue specific patterning of ACC oxidase and ACC synthase gene expression shifted from the embryo to the silique wall. These data demonstrate how low O2 modulates the activity and expression of the ethylene biosynthetic pathway during seed development in Arabidopsis. PMID- 12092616 TI - Can we trust the doctors? Parents' groups say that voluntary guidelines aren't enough. PMID- 12092617 TI - Geriatric medicine. PMID- 12092618 TI - Capsule endoscopy. PMID- 12092619 TI - The unraveling of health insurance. PMID- 12092620 TI - Welfare aspects of FMD discussed at BVA AWF seminar. PMID- 12092621 TI - Influence of the design of facilities at auction markets and animal handling procedures on bruising in cattle. AB - The movements of cattle at 12 livestock auction markets were observed to determine the possible causes of trauma leading to carcase bruising. Design faults included right-angled bends in races, dead ends, flooring with insufficient slope or grip, and steps. Sliding gates were often misused for goading cattle. Projecting fittings and square-edged corners were potentially injurious; conversely, rounded posts and curved races assisted the flow of cattle with minimal impacts. At all the markets, some cattle were hit directly and poked with wooden sticks, and the variations in their use could partially explain the differences between the markets in the prevalence of carcase bruising. Most harder hits were directed at less valuable parts of the body, such as the spine, hips and shoulders, and a survey of bruising at the abattoir showed that these were the areas with most bruising. In this survey of 48,926 carcases, the overall level of commercially significant bruising of 4.1 per cent was lower than the 6.5 per cent found in a previous survey. Carcases of cattle from markets had a greater incidence and severity of bruising (P<0.001) than those arriving directly from farms or dealers. Carcase bruising in young bulls was less (P<0.001) than in heifers and steers. PMID- 12092622 TI - Embryo collection in prepubertal gilts and attempts to develop an improved embryo transfer technique. AB - Prepubertal gilts were treated with 1,500 iu equine chorionic gonadotrophin, followed 72 hours later by 500 iu human chorionic gonadotrophin (hCG), and inseminated 36 and 48 hours later. Embryos were collected at slaughter 168 hours after the hCG treatment. Blastocysts classified as 'good' or 'fair' were transferred to synchronised recipients, either by conventional surgical means or by a 'semi-endoscopic' approach, and the recipients were slaughtered four weeks later. Of 238 donor gilts, 98.4 per cent had responded with a mean (se) 23.5 (1.0) ovulations and 19.1 (1.0) ova or embryos, of which 47 per cent were considered morphologically intact and transferable. The large proportion of non transferable embryos was not associated with the age or weight of the gilts, the season or with their housing conditions. Conventional surgical transfer of 15 to 20 (mean 17.4) blastocysts to synchronised recipients yielded 88 percent (14 of 16) pregnancies with between seven and 14 (mean 8.2) viable fetuses, and an embryo survival rate of 47 per cent in the pregnant recipients and 41 per cent in all the recipients. The corresponding data for the semi-endoscopic transfers were 16 to 20 (mean 17.7) blastocysts transferred, 47 per cent (eight of 17) pregnancies, four to 12 (mean 7.3) viable fetuses per pregnant recipient and an embryo survival rate of 41 per cent in the pregnant recipients and 19 per cent in all the recipients. Significantly fewer of these recipients became pregnant and a significantly smaller proportion of the embryos survived (P<0.05). PMID- 12092623 TI - Treatment of temporary upward fixation of the patella in an alpaca by abrasion trochleoplasty and imbrication of the medial retinacular fascia. AB - A two-year-old female alpaca suddenly became lame on its right hindlimb. Eight days later clinical and radiographic examinations showed that the patella had become temporarily fixed within the supratrochlear notch of the femur. Under general anaesthesia an abrasion trochleoplasty followed by fascial imbrication was carried out. After two weeks in supporting slings, the animal put full weight on the leg, and six months after the surgery it showed no signs of lameness or recurrence of the upward fixation. PMID- 12092624 TI - Ringworm caused by Trichophyton verrucosum--an emerging problem in sheep flocks. PMID- 12092625 TI - Echinococcus multilocularis infection in red foxes in italy. PMID- 12092626 TI - Multiresistant Salmonella enterica serovar Newport. PMID- 12092627 TI - Welfare of horses during transport for slaughter. PMID- 12092628 TI - Sleeping disease in trout. PMID- 12092629 TI - National biosecurity against infectious diseases. PMID- 12092630 TI - Roles of the RCVS and BVA. PMID- 12092631 TI - Drug addiction and the veterinary profession. PMID- 12092632 TI - Drovers' return? PMID- 12092633 TI - Homocysteine and C-reactive protein levels in haemodialysis patients. AB - BACKGROUND: Mild to moderate hyperhomocysteinemia is very common among patients undergoing haemodialysis. There is sufficient evidence that hyperhomocysteinemia is an independent risk factor for cardiovascular and or atheromatous disease in end stage renal failure patients. Vitamin supplementation such as vitamin B6, B12 or folate has been proposed to correct this metabolic disturbance and it is to be proved if this intervention benefit these patients, but there is no agreement whether oral folate supplementation is capable to normalize homocysteine levels in end stage renal failure patients. METHODS: In 53 patients, undergoing haemodialysis, homocysteine levels (Hcy), folate, vitamin B12, ferritin and C reactive protein (CRP) were estimated before and after dialysis, without folate supplementation. Thirty voluntary blood donors were used as controls to compare homocysteine levels. After four weeks of oral folate supplementation (10 mg/24 hours) the levels of homocysteine, serum folate and intra-erythrocyte folate were estimated again. Eighteen months later the survival rate of our patients was recorded and analyzed in relation to Hcy and CRP levels. RESULTS: The results showed that haemodialysis patients exhibited, almost, fourfold higher homocysteine levels than controls (27.39 +/- 11.54 vs 7.38 +/- 3.5, t = -8.2, p = 0.000000). Folate levels, vitamin B12 and CRP increase significantly after haemodialysis where as homocysteine levels decrease (Hcy1 vs. Hcy2: z = 2.08, p = 0.03). Fourteen (14) patients suffered from coronary heart disease (CHD) and they exhibited the higher levels of homocysteine (Hcy1 vs. CHD: z = -3.4, p = 0.0006). All estimations performed revealed a negative correlation between homocysteine levels and plasma or intra-erythrocyte folate. No other variable exhibited any significant influence upon homocysteine levels. After folate supplementation homocysteine levels in the whole number of patients were unchanged (Hcy(before) vs. Hcy(after): 27.39 +/- 11.54 vs. 26.95 +/- 8.22, z = 0.3, p = 0.7, NS). When patients with homocysteine levels higher than 24 micromol/L were selected, a significant decrease was observed (34.77 +/- 9.32 vs. 30.0 +/- 8.05, z = 2.09, p = 0.02). Forty-two patients were treated with erythropoietin for their anemia and we found a positive correlation between C-reactive protein levels and rhu-Epo dose (CRP vs. Epo: r = 0.45, p = 0.002). Homocysteine levels did not exhibit any significant influence upon short-term survival (U = -0.37, p = 0.3, NS) where as CRP levels exhibit a significant influence upon short-term survival (U = 2.15, p = 0.005). CONCLUSIONS: Homocysteine levels in haemodialysis patients are fourfold higher than healthy controls. Folate, vitamin B12 and CRP increases significantly after dialysis. Patients with coronary heart disease exhibit the highest levels of homocysteine. The homocysteine levels are inversely related with the folate levels. The exogenous folate supplementation increase the serum folate levels but decreases homocysteine only in patients with higher than mild hyperhomocysteinemia. Hcy doesn't exert any significant effect upon the short term survival of the haemodialysis patients but CRP level is a god predictor of the short-term survival of these patients. PMID- 12092634 TI - Randomized trial of a combination of natural products (cernitin, saw palmetto, B sitosterol, vitamin E) on symptoms of benign prostatic hyperplasia (BPH). AB - Because benign prostatic hyperplasia (BPH) is relatively common, it is important to discover safe and effective means to treat this often debilitating perturbation. Accordingly, we examined the effectiveness of a combination of natural products (cernitin, saw palmetto, B-sitosterol, vitamin E) in treating symptoms of BPH. We undertook a randomized, placebo-controlled, double-blind study. Patients were enrolled from 3 urological practices in the USA. 144 subjects were randomized for study. 17 subjects eventually withdrew, leaving 70 patients in the test group and 57 in the placebo group to complete the study. Inclusion criteria consisted of a diagnosis of BPH, no evidence of cancer, and a maximal urinary flow rate between 5 and 15 ml/second. Patients received either placebo or the combined natural products for 3 months. Evaluations were performed via the American Urological Association (AUA) Symptom Index score, urinary flow rate, PSA measurement, and residual bladder volume. Nocturia showed a markedly significant decrease in severity in patients receiving the combined natural products compared to those taking placebo (p < 0.001). Daytime frequency was also lessened significantly (p < 0.04). When the average individual total AUA Symptom Index score in the test group was compared to that in the placebo group at the end of the study, the difference proved highly significant (p < 0.014). PSA measurements, maximal and average urinary flow rates, and residual volumes showed no statistically significant differences. When taken for 3 months, a combination of natural products (cernitin, saw palmetto, B-sitosterol, vitamin E) compared to placebo can significantly lessen nocturia and frequency and diminish overall symptomatology of BPH as indicated by an improvement in the total AUA Symptom Index score. The combination of natural products caused no significant adverse side effects. PMID- 12092635 TI - Spectrum of renal diseases in the elderly: single center experience from a developing country. AB - From November 1998 to March 2000, two hundred patients over the age of 60 years (Elderly) with clinical renal disease were studied. 144 patients were between ages of 60-69 years, 46 between 70-79 years and 10 were above 80 years. The elderly patients (Male 165; Female 35) with renal disease constituted 11% (200/1816) of the total nephrology consultation during the study period. The clinical presentation included chronic renal failure (42.5%); acute renal failure (28%); nephrotic syndrome (14.5%); acute glomerulonephritis (7.5%); renal vascular disease (5%) and renal cystic disease (2.5%). Diabetic nephropathy, obstructive uropathy and hypertensive nephrosclerosis were the major causes of CRF, accounting for 80% of total CRF in the elderly. Chronic glomerulonephritis and chronic pyelonephritis (CPN) were less common and etiology of CRF was uncertain in 5.9% of cases. However, diabetic nephropathy was the commonest (49.4%) cause of chronic renal failure. We did not see a single case of ischemic nephropathy causing CRF in the present study. Prerenal ARF, obstructive uropathy and sepsis were contributing factors for ARF in 82% of the cases. Volume depletion due to gastrointestinal fluid loss and urinary tract obstruction on account of enlarged prostate were the leading causes of ARF in 20 (35.7%) and 8 (14.3%) cases respectively. Sepsis with or without multiorgan failure was the major (46.7%) cause of mortality in patients with ARF and overall mortality was 26.8%. The commonest (31%) cause of nephrotic syndrome was the idiopathic membranous nephropathy. Diabetic nephropathy related to type-2 diabetes mellitus was the second most common (24.1%) cause of nephrotic syndrome. Diffuse endocapillary proliferative GN of post infectious etiology was the commonest (73.3%) type of acute GN in our elderly patients. Renal cystic diseases were noted in 5 (ADPKD 3; Simple cyst-2) patients. Thus, overall spectrum of renal disease in our elderly patients is similar to that of developed nations except in two ways: (i) Endocapillary proliferative GN of post infectious origin was the commonest type of acute GN and (ii) Rarity or absence of ischemic nephropathy and atherosclerotic renal artery occlusive disease. PMID- 12092637 TI - Screening for prostate cancer. AB - Carcinoma of the prostate gland is the second most common cancer among men with an age-adjusted incidence of 635 cases per 100,000 men aged 65 and older. While there are several proven methods for detecting prostate cancer, debate continues as to the best way to detect it early as well as who should receive particular screening. There are differing opinions as to proven benefit even when cancer is detected. Fortunately, newer methods continue to be developed that will hopefully reduce false positive detection rates while insuring an adequate level of screening protection. PMID- 12092636 TI - Molecular physiology of urinary concentration defect in elderly population. AB - It is estimated that by the year 2050 one in five Americans will be 65 years or older. This mandates the growing need for clinical and basic research in the field of geriatric medicine to understand age-related maladies. The most prominent abnormality in renal function in the aging population is the inability to handle water, frequently resulting in hypo- or hyperosmolar states, and the associated electrolyte imbalances. During the past decade, thanks to the advent of powerful molecular techniques, rapid strides have been made in the approaches employed to understand and dissect the physiology of renal function in general and the urinary concentration mechanism in particular. Using an integrated approach of clinical observations, experimental model systems, molecular analysis, and functional genomics, a more comprehensive picture of the interplay of physiological systems in the genesis of urinary concentration defect in the elderly is beginning to emerge. Much remains to be deciphered regarding the complex interactions between the role of environment, genetics, diet, pharmacological agents and the general effects of aging on kidney function. The emerging importance of socio-economic and quality of life issues surrounding geriatric medicine encourage public and private support and funding for research in the area of age-related diseases, especially as they are related to the kidney. PMID- 12092638 TI - Comparisons of hemodialysis and CAPD in patients over 65 years of age: a meta analysis. AB - This meta-analysis had the aim of studying the available studies on comparison between Hemodialysis and Peritoneal Dialysis in the elderly. The final objective was to reach, if possible, evidence for potential differences. In the case that no differences could be demonstrated, contribute to accept that HD and PD are similar techniques to be offered to elderly people requiring dialysis. The question formulated was this: Do we have adequately contrasted data on results for survival, hospitalization rate, quality of life and morbidity on hemodialysis and peritoneal dialysis in the elderly (more than 65 years old)? As data sources we selected eight papers that compared the general results of these two dialysis techniques. Different elements were considered in this selection because none reached the two first levels in the hierarchy of sources of evidence, and only two reached the third level--that of prospective studies; this is because an oral presentation of data has been included in a meta-analysis. Another four papers- uni- or multicenter retrospective studies compared the results obtained with PD and HD. The remaining two papers--reports from nationwide registries that compare of mortality rates, adjusted for co-morbid conditions and age, present specific results on groups of elderly patients. Three papers compare particular aspects of the two techniques, including nutritional status, psychiatric and psychosocial aspects and rehabilitation, in this case comparing PD with home hemodialysis patients. Finally, we have included the opinions of healthy elderly people on dialysis issues. This meta-analysis of these different studies suggests that the mortality and hospitalization rate of elderly people treated by PD is similar to that of similar people treated by HD. In consequence, we have no reasons to select either therapy on behalf of the patient. The nephrologist should consider and inform the patient and family about the relative advantages and disadvantages of both techniques and tailor dialysis technique choice to the specific individual to assure the best results. Local circumstances should also be considered. PMID- 12092640 TI - Geriatric nephrology and urology literature. Vol. 33 #2. PMID- 12092639 TI - Clinical pharmacology-physiology conference. A chrono-therapeutic approach to the treatment of hypertension. PMID- 12092642 TI - Efficacy and outcome of surgical intervention in patients with nephrolithiasis and chronic renal failure. AB - AIM: To prospectively evaluate the efficacy and outcome of surgical intervention in patients with renal stones and chronic renal insufficiency. METHODS: The study was carried out from January 1999 till January 2001. Only patients with chronic renal failure without medical renal disease were taken up for study. All patients were subjected to an ultrasound assessment of the kidney, ureter and bladder. In case of obstructed and infected systems a preliminary percutaneous nephrostomy was carried out. After correction of dyselectrolytemia, acid base imbalance and dialysis (if indicated) patients were subjected to surgical intervention (open surgery or percutaneous nephrolithotomy). ESWL was offered for stones persisting after surgery. The stone burden, composition, therapeutic procedures required to render patients stone free were assessed. The outcome of stone removal on renal function was also evaluated by serial renal dynamic scans and creatinine clearance estimations. RESULTS: Out of 90 patients operated for staghorn or calyceal calculi, complete follow up data was available in 70. Pyelo nephrolithotomy and percutaneous nephrolithotomy was carried out in 63 and 7 patients respectively. Out of 15 patients with residual stones ESWL was successfully performed in 9 cases. Mixed, calcium oxalate monohydrate, calcium oxalate dihydrate, and struvite stones were encountered in 48%, 14%, 17% and 21% respectively. The average pre operative serum creatinine was 4.76 (1.9-16) mg%. The maximum duration of follow up was 9 months. By the 9th post operative month the average fall in serum creatinine values was 1.53 mg/dl (32%) and the average functional improvement by renal dynamic scans stood at 20.665%. 41 patients were saved from further dialysis. CONCLUSION: Patients with mild to moderate renal failure showed maximal improvement in renal function forestalling or reducing the need for dialysis/renal replacement therapy. PMID- 12092641 TI - Common conditions of the aging male: erectile dysfunction, benign prostatic hyperplasia, cardiovascular disease and depression. AB - With increasing life expectancy, medical profession will be faced with the task of ensuring that the large, aging population remains healthy and vital despite the face of increasing healthcare costs. Naturally, urology as a specialty is concerned with important geriatric issues. This article will focus specifically on the aging male population and the health problems that most frequently plague them. Four major, non-cancer, disease states have been identified that adversely effect males over the age of 50. These conditions are: erectile dysfunction (ED), benign prostatic hyperplasia (BPH), cardiovascular disease (CVD) and depression. A literature search of PubMed was conducted using the key words ED, BPH, CVD and depression as well as ADAM (Androgen Decline in the Aging Male) and quality of life (QoL). NIH and WHO conference proceedings and publications were also referenced to insure detail and accuracy of data. Information was then organized and correlated in order to provide a detailed description of the key conditions and their interrelatedness. The spectrum of research performed thus far regarding this topic has done little to investigate the effects, causes and correlations between these conditions. Research has been done linking two or three of these conditions; however, there remains to be information discussing the four disease states in terms of their possible cause and effect relationships or the effectiveness of parallel, multi-disciplinary approach to their therapy. This report calls attention to the benefits of viewing and researching the above mentioned conditions as possibly interrelated, as opposed to the traditional view of them as separate, unrelated and independently treatable disease states. ED, BPH, CVD and depression are all common conditions that accompany aging and negatively impact QoL. They almost always develop with age and precipitate considerable morbidity and may even result in mortality. Furthermore, the presentation of one condition may correlate with the development of another. The inter-relation of these conditions, as evident from their underlying similarities, cause-and-effects relationships and therapeutic consequences, should be enough to warrant a multidisciplinary approach to their research. This approach, combined with careful choice of therapy, parallel and singular, will help providers reach their goal to keep patients healthy, and more importantly happy, late into their life, thus realizing the concept of "successful aging". PMID- 12092644 TI - Cost-reductive retroperitoneal excision of large adrenal pseudocyst: a case report and review of the literature. AB - We report a case of a non-functioning cystic adrenal mass. It was detected on an abdominal ultrasound carried out for right flank pain in a 45-year-old lady. Biochemical parameters like 24-hour urinary free cortisol, catecholamines and serum potassium level were normal. Retroperitoneoscopic excision of the large cystic mass was done by using our cost-reductive technique as previously described. Histopathology turned out to be a pseudocyst with wall fibrosis and calcification. PMID- 12092643 TI - The expression of cytoskeletal proteins (alpha-SMA, vimentin, desmin) in kidney tissue: a comparison of fetal, normal kidneys, and glomerulonephritis. AB - BACKGROUND: The aim of the study is a comparison of the expression of cytoskeletal proteins, alpha smooth muscle actin (alpha-SMA), vimentin, and desmin in fetal, normal kidney and proliferative (diffuse proliferative and membranoproliferative glomerulonephritis) and nonproliferative (membranous glomerulonephritis) glomerulonephritis. METHODS: We have studied the expression of cytoskeletal proteins (alpha-SMA, vimentin, desmin) in the paraffin embedded tissue sections from the kidneys of 10 normal kidney (adults and infants), 13 fetal kidney, 12 membranous glomerulonephritis (MGN), 8 membranoproliferative glomerulonephritis (MPGN), 8 diffuse proliferative glomerulonephritis (DPGN). Interstitial and glomerular positive stainings were evaluated. RESULTS: Vimentin expression was similar in normal infant and adult kidneys with positive staining in glomeruli and negative staining in interstitium. In fetal kidneys, glomerular mesangial and epithelial cells and blastematous areas showed positive reactivity with vimentin. Alpha-SMA staining was different among the groups. In fetal kidney, alpha-SMA expression was found in glomerular mesangial cells and blastematous areas. Alpha-SMA staining was positive in peritubular area and glomerular mesangial cells in infant kidney. In adult kidneys, glomerular staining with alpha-SMA disappeared but peritubular positivity continued. Interstitial staining with alpha-SMA was positive in fibrotic areas of proliferative (MPGN, DPGN) and non-proliferative (MGN) glomerulonephritis, but positive glomerular staining with alpha-SMA was found only proliferative glomerulonephritis. Desmin expression was negative in all groups. CONCLUSIONS: Desmin is not expressed in early stages of kidney growth, infant and adult kidneys, and proliferative and nonproliferative glomerulonephritis. Interstitial staining of vimentin in the diseased kidney tissues revealed increased fibrosis. Alpha-SMA revealed important differences in different stages of nephrogenesis. Glomerular mesangial staining with alpha-SMA in developing (fetal and infant kidneys) and proliferative glomerulonephritis suggest that it may be a marker of proliferation. In addition, it shows myofibroblastic differentiation in interstitium in diseased kidneys. PMID- 12092645 TI - Acute renal failure due to bilateral matrix renal calculi--a diagnostic dilemma. AB - We report an unusual case of acute renal failure caused by bilateral matrix calculi. The difficulties in diagnosing the cause of renal failure and the management of the case are discussed. Matrix calculi are rare causes of acute renal failure. Being radiolucent they can be elusive and systematic investigation and a diligent search are required for correct diagnosis and appropriate treatment. PMID- 12092646 TI - Ondansetron versus diclofenac sodium in the treatment of acute ureteral colic: a double blind controlled trial. AB - The aim of this study is to compare the effectiveness of the 5-HT3 antagonist, ondansetron and a non-steroidal anti-inflammatory agent, diclofenac sodium, as a pain reliever in the treatment of acute ureteral colic. Sixty four patients with severe or moderate pain who were clinically diagnosed as having ureteral colic associated with microscopic or gross hematuria were included in the study. Thirty three patients were administered ondansetron and 31 patients were administered diclofenac sodium. Exclusion critera were known kidney or liver disease causing dysfunction, known hypersensitivity to ondansetron or diclofenac sodium, pregnancy, lactation, duodenal ulcer or bleeding. After pain assessment with a verbal scale and a visual analog scale (VAS), we randomized patients and administered 8 mg ondansetron intravenously to 33 patients and 75 mg diclofenac sodium intramuscularly to 31 patients and pain scores were recorded every 15 minutes. If significant pain relief was not achieved within 60 minutes, i.v. meperidine was given as rescue pain medication. Ondansetron was effective as a primary pain reliever in 14 (42.4%) patients, whereas 19 patients required additional medication. Diclofenac sodium was effective as a primary pain reliever in 24 (77.4%) patients, whereas 7 patients required additional medication. Ondansetron was not superior to diclofenac sodium in relieving pain in patients with acute ureteral colic. PMID- 12092647 TI - Stone necklace of urinary tract presenting as renal failure: one stage management. AB - We describe three cases with bilateral extensive involvement of both upper and lower urinary tract with calculi presenting in renal failure. The management of these patients in a single operative session and rendering them stone free is discussed. Modern endourologic techniques have made it possible to treat patients with such an extensive involvement of the urinary tract with stone disease with minimum morbidity. PMID- 12092648 TI - Laser endoscopic incision: a viable alternative to treat adult ureterocele. AB - AIM: To prospectively evaluate the safety. efficacy, feasibility and outcome of Nd-Yag laser endoscopic incision in adult symptomatic patients with intravesical uretroceles. METHODS: We present our experience with the use of Nd-Yag laser for endoscopic incision of the adult ureterocele in a group of five symptomatic adult patients. RESULTS: All the five patients showed excellent decompression with relief from flank pain and hydronephrosis and none demonstrated a post-op vesicoureteric reflux (VUR). CONCLUSION: Laser endoscopic management of intravesical ureteroceles is new emerging modality, which is being increasingly employed to decompress ureteroceles. Laser endoscopic incision is a precise and bloodless procedure enabling a clean accurate controlled cut through the endoscope as opposed to the traditional bugbee electrode. PMID- 12092649 TI - Hydatid disease of the urinary tract: review of the management of 9 cases. AB - The aim of this study is to review 9 cases with urinary tract hydatid disease in terms of symptoms, findings, laboratory tests, radiological findings and treatment modalities. There were 7 males and 2 females with a mean age of 33.6 years (range from 7 to 67 years). In 6 patients hydatid cyst was located in the kidney (1 involved the liver), in 2 the cysts were in the paravesical and retrovesical region (1 coexisted bladder tumor) and 1 the cyst was located adrenal gland. The investigations included urinalysis, eosinophil count, Casoni skin test, indirect haemagglutination test (IHA), transabdominal ultrasonography (TAUS), intravenous urography (IVU) and computed tomography (CT). All patients underwent various surgical procedures and were followed-up for an average of 3 years (range: 1 month to 5 years) period in terms of complications and recurrence rates. The most common symptom was lumbar or abdominal pain. Eosinophilia was seen in 4 patients (44.4%), IHA positivity in 3 patients (33.3%) and Casoni skin test were positive in 3 patients (33.3%). The most diagnostic method of radiological investigations was abdominal pelvic CT (100%). After surgical treatment, in mean a 3 year (range: 1 month to 5 years) follow-up period, no complication and recurrence were seen. Hydatid disease of the urinary tract is relatively uncommon and is likely to cause considerable diagnostic difficulties for clinicians and radiologists: therefore, it should be born in mind in the differential diagnosis of space-occupying lesions of the urinary tract. PMID- 12092650 TI - Estimating the effectiveness of various methods of evacuation of kidney stones, on the basis of data obtained on percentage of "stone free" and recurrent stone formation. AB - Using the materials of more than 200 scientific papers of recent years in this review it has been carried out analysis of the effectiveness and also discussed the merits and demerits of various methods of treatment of urinary stone diseases (USD). With this purpose, for the first time it has been carried out analysis of treatment quality of USD using more important quantitative parameters of treatment efficacy: 1--percentage of "stone free"; 2--percentage of recurrent stone formation. It has been emphasized on the higher effectiveness of treatment in combination of the method of ESWL with the methods of ascending litholysis. It was shown that in order to solve problems such as etiology and pathogenesis of the disease, elaboration of quantitative risk factors of the disease, and search for effective measures on pro- and metaphylaxis of USD, and also elaboration of various (including therapeutics) medium and methods for complete and rapid evacuation of stones, it is needed to carry out intensively fundamental and applied researches in the field of urinary stone diseases. PMID- 12092651 TI - Contracted bladder secondary to eosinophilic cystitis. PMID- 12092652 TI - Malignant paraganglioma of the urinary bladder. A case report. AB - Paraganglioma (pheochromocytoma) is probably the most fascinating of all tumors as it can present with a wide range of clinical manifestations. Paraganglioma of the urinary bladder is one of the rare tumors and constitute less than 10% of all bladder tumors. The common presentation of paraganglioma of the urinary bladder is painless haematuria, headache, palpitation and anxiety. Malignancy is uncommon and no histological feature is characteristic of malignancy. Invasion of adjacent organs or metastasis to the lymph nodes is the only criteria to suggest malignancy. In the presence of proven metastasis radical cystectomy with pelvic lymphadenectomy is recommended. In the present case the tumor was arising from anterior wall of the bladder away from the bladder neck and that in a large capacity bladder it was possible to get a wide margin for partial cystectomy along with bilateral pelvic lymphadenectomy without violating the oncological principles. At a follow up of two years there is no evidence of recurrence of tumor. PMID- 12092653 TI - A very unusual case of chronic abdominal pain caused by a giant bladder stone formed on a Foley catheter left in bladder. PMID- 12092654 TI - Endoscopic management of an unusual foreign body in the urinary bladder leading to intractable symptoms. AB - A 70-year old female patient presented with intractable lower abdominal pain and recurrent urinary tract infection following an endoscopic bladder neck suspension. Investigations revealed it to be a case of suture and pledget migration leading to foreign body granuloma in urinary bladder. It is being reported as an uncommon complication of endoscopic bladder neck suspension. An early endoscopic evaluation should be carried out in cases of unexplained lower urinary tract symptoms following any surgical procedure for incontinence. It is also appropriate to retrieve these foreign bodies endourologically without resorting to open surgery and thus extending safe, comfortable, and short postoperative course with good long term results. PMID- 12092655 TI - AgNOR/P53 expression compared with different grades in bladder carcinoma. AB - OBJECTIVE: The aim of this study is to assess the relation between silver strained nucleolar organizer regions (AgNOR)/P53 expression and urothelial neoplasm of the bladder which graded according to the WHO/ISUP tumor grading system. METHODS: Transurethral resection materials (TUR) and cystectomy specimens from 52 patients with bladder urothelial carcinomas were stained by AgNORs and analyzed immunohistochemically using an avidinbiotin peroxidase method for p53 staining and also silver for AgNORs. RESULTS: Nuclear p53 accumulation was determined in two of 5 LMP cases (%40), in 10 of 22 LGC (%45.4), and in 8 of 25 HGC (%32). Positivity of p53 accumulation was observed in 20 (%57.7) of all cases. There was no statistically significant relationship between AgNOR score of p53(+) and p53(-) cases. There was no correlation between LGC and LMP, also between LGC and HGC (p > 0.05), but a meaningful relationship was found between LMP and HGC (p = 0.03). Association between P53 and grade, AgNOR value and grade, p53 and AgNOR was found statistically insignificant. CONCLUSIONS: Although we found a statistically significant correlation between AgNOR count of LMP and HGC, this study did not show a satisfying correlation between AgNOR, p53 and any grade of urothelial carcinoma. PMID- 12092656 TI - Radical cystectomy versus alternative treatments for muscle-confined bladder cancer. AB - Of the 100 patients with muscle-confined transitional cell cancer of the bladder and ASA score < or = 3, 59 underwent radical cystectomy (RC) and 41 received non cystectomy alternative treatments (AT). Median follow-up was 30.8 and 30.5 months in RC and in AT groups, respectively. Disease-free and overall survivals were significantly longer in RC group than AT group. Salvage cystectomy was required in approximately 25% of the patients who received AT. AT was associated with higher rate of cancer-related morbidity and cancer progression than RC. Every patient with invasive bladder cancer should be given a chance for cystectomy. PMID- 12092657 TI - K-RAS mutation in transitional cell carcinoma of urinary bladder. AB - In the present study it was aimed to investigate the frequency of K-RAS mutation in the human bladder transitional cell carcinoma. For this purpose, tissue specimens obtained from the patients with bladder tumors. Genomic DNAs were isolated and then PCR-SSCP analysis of K-RAS gene were performed. A heterozygous deleted mutation was detected in K-RAS oncogene (exon 2) in agarose gel electrophoresis in one patient and point or substitution mutations are detected using single strand conformational polymorphism (SSCP) in other different patients with bladder cancer (4/14). In conclusion, the frequency of K-RAS mutation is not rare and the role of this mutation in oncogenesis and in infiltration of the urinary bladder wall needs to be confirmed in a larger study. PMID- 12092658 TI - Nephrogenic adenoma of the prostatic urethra in a patient treated with ESWL for a lower ureteral stone. AB - We present a case of prostatic urethral nephrogenic adenoma incidentally diagnosed in a patient treated with ESWL for ureteral stone disease, and we comment on this unusual urothelial lesion. PMID- 12092659 TI - Colovesical fistula an unusual complication of cytotoxic therapy in a case of non Hodgkin's lymphoma. AB - A 65-year old man, a known case of non-Hodgkin's lymphoma of base of the tongue and epiglottis presented with complaints of pneumaturia and faecaluria. He had received the first cycle of cytotoxic therapy (CHOP-regimen). At the end of the cycle he developed febrile neutropenia (circulating granulocyte count <1500/mm3). Cystogram showed air in the bladder area and a fistulous communication to a cavity behind the bladder. CT-scan showed air in the bladder, a fistulous communication between the sigmoid colon and bladder along with an intervening small abscess cavity. On exploration a fistulous communication between the sigmoid and bladder along with an intervening small abscess cavity was found. Resection of involved portion of sigmoid and end to end anastomosis along with a diverting colostomy was done. The bladder was closed in two layers with an omental interposition between it and the sigmoid along with a suprapubic cystostomy. The histopathology demonstrated only inflammatory response without any evidence of malignancy or diverticular disease. PMID- 12092660 TI - The incidence of rectal bleeding following three-dimensional conformal radiotherapy of prostatic cancer. AB - This study evaluated the incidence of rectal bleeding in patients with prostatic carcinoma treated with three-dimensional conformal radiotherapy (3D-CRT). A total of 32 patients received mean 68.87 Gy 3D-CRT, the mean follow up time was 23.68 months. There was not statistically significant difference between the groups of > or = 70 Gy and < 70 Gy in terms of rectal bleeding (p > 0.05). but there was statistically significant difference between the groups of who have previous history and finding of rectal bleeding and who have not previous history and finding of rectal bleeding (p < 0.013). Rectal bleeding of the patients had significant resolution of their symptoms with conservative therapies. PMID- 12092661 TI - A phase II trial with new triptorelin sustained release formulations in prostatic carcinoma. AB - OBJECTIVES: The objectives were to assess if a single intramuscular (i.m.) injection of the GnRH agonist triptorelin, as pamoate Sustained Release (RS) 11.25 mg, was able to induce pharmacological castration and to maintain the plasma testosterone levels in the castrate range (< 1.735 nmol/l) up to 3 months in prostatic carcinoma. METHODS: Two different formulations of triptorelin pamoate 11.25 mg were assessed in 2 groups of 10 patients suffering from prostatic carcinoma. Each patient received one i.m. injection of triptorelin pamoate SR 11.25 mg. Triptorelin and testosterone levels were measured over 3 months. Pain, micturition difficulties, performance status, local and general tolerance, and the occurrence of adverse events were evaluated. RESULTS: Both formulations were able to induce castration levels (<1.735 nmol/l) of testosterone within 3 to 4 weeks post-injection, and to maintain levels below 1.735 nmol/l till the end of 3rd month. The bioavailability of one formulation (DLGSD-3-95-21) tended to be greater. This may explain the quicker onset of castration and the slight better maintenance of low testosterone levels during the 3rd month observed with this formulation. In terms of clinical end-points, the local tolerance of both formulations was excellent. No serious adverse events were recorded except transient hot flushes in 2 cases and slight bone pain in one. CONCLUSION: Triptorelin pamoate 11.25 mg given in microgranules is a 3-month sustained-release administration form which appears to be safe and effective in advanced prostatic carcinoma. Based on the findings of this study, the formulation with greater bioavailability (DLGSD-3-95-21) was selected as formulation of choice to be used for clinical treatments and further clinical investigation. PMID- 12092662 TI - Successful microsurgical replantation of an amputated penis. AB - Traumatic amputation of the penis is a rare condition. The paper presents a 22 year old man who suffered a complete penile amputation in a result of an unfortunate accident. Microsurgical replantation of the penis was performed in the case with a complete return of sensation, sexual function and physiologic miction. Microsurgical replantation is the treatment of choice for this injury. PMID- 12092663 TI - Sildenafil test: changes in the diagnostic and therapeutic management of erectile dysfunction. AB - OBJECTIVE: To assess the efficacy of sildenafil as a first-step diagnostic and therapeutic tool for erectile dysfunction (ED) and to evaluate the consequent changes in the management of male sexual insufficiency. MATERIALS AND METHODS: Sildenafil in titrating doses up to 100 mg was prescribed to 50 men presenting to a sexual dysfunction clinic with medically documented ED. They had not undergone any specific diagnostic test before starting sildenafil. RESULTS: Of the 50 men, 24 (48%) responded to sildenafil. Of these, 8 (33.3%) responded to 50 mg and 16 (66.7%) to 100 mg of sildenafil. Of the responders, 9 representing 18% of all studied men were discharged achieving spontaneous erections in a mean follow-up of 5.3 months. Men with no medical history, men with hypertension and men with mild coronary artery disease responded better. CONCLUSIONS: The sildenafil test revealed that 48% of men responded to this therapy with no requirement for more invasive tests and that 18% of men required no further treatment at all. In addition this test reduced the overall cost of the diagnostic investigation. It is proposed that the sildenafil test should be used in cases with no significant medical history or in men with hypertension or mild coronary artery disease although almost all men with ED could be categorized as sildenafil-responders or sildenafil-resistant. It is also suggested that the sildenafil test would result in the ability for more men with ED to be managed exclusively in the primary care sector. PMID- 12092665 TI - Urethral stone presenting as a stop valve--a rare complication of balanitis xerotica obliterans. AB - Balanitis xerotic obliterans (BXO) is the genital subcategory of lichen sclerosis et atrophicus. The association of BXO with urethral stone causing interruption of the urinary stream and voiding by manual displacement of the urethral stone has not been described before. We describe one such case of a young boy with BXO and urethral stone who voided by manually displacing the stone for over a year. The case is reported to emphasize the ingenuity of the patient in continuing to void for over a year despite the association of the impacted urethral stone with urethral stricture and BXO. PMID- 12092664 TI - Relaxation degree: a new concept in erectile dysfunction. AB - PURPOSE: To investigate the clinical application of the new parameter' Relaxation Degree. PATIENTS AND METHODS: Chart reviews of 150 patients were evaluated retrospectively of whom 96 had been diagnosed as pure cavernoocclusive dysfunction (COD) by using corpus cavernosum electromyography (CC-EMG) and cavernosometry and of whom 54 had been diagnosed as psychogenic erectile dysfunction by normal CC-EMG; cavernosometry and pulse doppler scanning. Relaxation degree was defined as the percent decrease in the amplitudes of the electrical activity of corpus cavernosum (EACC). RESULTS: Mean relaxation degree measurements were found as 66.5%, 66.28% and 16% in minimal, moderate and severe COD groups respectively. This value was 80% in psychogenic erectile dysfunction group. Statistical comparisons were found significant between minimal and severe and moderate and severe COD groups. All three organic erectile dysfunction groups were also compared with psychogenic erectile dysfunction group where differences were found significant. If the normal relaxation degree was accepted as more than 80%, COD could be diagnosed with 93% specificity and 98% sensitivity without performing cavernosometry. CONCLUSION: Relaxation degree parameter could be used in diagnosis of COD showing the percent relaxation ability of the cavernous smooth muscle as a non invasive method when compared with cavernosometry. PMID- 12092666 TI - Liposarcoma of the spermatic cord. AB - Primary malignant tumors of the spermatic cord are rare. We report a case of paratesticular liposarcoma and discuss the prognosis of this neoplasm and its therapeutic implications. PMID- 12092667 TI - Accumulation of toxic products degradation of kynurenine in hemodialyzed patients. AB - In patients that developed a chronic renal failure the augmentation in tryptophan degradation is reflected in the increase in plasma metabolites of kynurenine pathway. Hemodialysis is one of therapeutic approaches that significantly reduce all plasma kynurenine metabolites in uremic patients. In spite of haemodialysis, plasma concentration of kynurenine, kynurenic acid, 3-hydroxykynurenine, anthranilic acid, xanthurenic acid and quinolinic acid were still elevated in uremic patients in comparison with healthy volunteers. These data shows significant disturbances in kynurenine metabolism in uremic patients. Accumulation of these substances in uremic blood is capable to account for certain uremic symptoms. PMID- 12092668 TI - The emerging role of adoption reunion registries: adoptee and birthparent views. AB - Since the 1980s, government-sponsored adoption reunion registries have emerged as key access points for individuals separated by adoption to secure information and initiate efforts to be reunited with their birth-families. Despite the prevalence of such registries, little information is available about their operational effectiveness. This article presents data from a survey of adult adoptees and birthparents who used the services of a statewide reunion registry. PMID- 12092669 TI - Bridging child welfare and juvenile justice: preventing unnecessary detention of foster children. AB - Gaps in service coordination between child welfare and other child-serving agencies are well documented. This article examines the gap between the child welfare and juvenile justice systems and discusses a program, Project Confirm, designed to reduce the problems associated with this gap. Project Confirm aims to improve cooperation between juvenile justice and child welfare agencies to prevent the unnecessary detention of arrested foster children in secure facilities. The program's design is outlined, and implementation statistics and government partner perceptions of the program in its first year of operations are provided. The article also identifies future challenges to implementation and discusses the broader implications of the program. In making this appeal for coordination and collaboration among public agencies, I'm not talking about pro forma integration efforts-I'm not talking about memoranda of understanding or top level reorganizations that have autonomous agencies vaguely reporting to a single box labeled "human services." Rather, I'm talking about the much more challenging objective of achieving real working partnerships at the front line. (Nelson, 1998, p. 6) PMID- 12092670 TI - The child health and illness profile-adolescent edition: assessing well-being in group homes or institutions. AB - The Child Health and Illness Profile-Adolescent Edition (CHIP-AE), a new standardized instrument, was administered to 63 adolescents living in group homes or institutions. Youth reported high levels of satisfaction with their physical health, resilience and problem-solving skills, and academic achievement. Youth reported low levels of self-esteem, emotional comfort and psychosocial stability, family involvement, and work performance. They also took more risks, had more threats to achievement, and had poorer peer influences than other youth. This study demonstrates the potential usefulness of the CHIP-AE as a tool for assessing the health and well-being of youth living in group homes and institutions. Suggestions for future use of the CHIP-AE are discussed. PMID- 12092671 TI - After adoption: dissolution or permanence? AB - Results are presented on the whereabouts of 516 adopted children, based on a random sample of children adopted from placement in New York City in 1996. Data from interviews with adoptive parents were augmented by information from adoption subsidy records and state child tracking files, as well as interviews with caregivers of children whose adoptive parents were deceased. There were few dissolutions, but postadoption service needs were many. PMID- 12092672 TI - Managed care redux. PMID- 12092673 TI - Consumer beliefs and health plan performance: it's not whether you are in an HMO but whether you think you are. AB - Surveys that rate how persons enrolled in HMOs and other types of health coverage feel about their health care are used to bolster claims that HMOs provide inferior quality care, providing justification for patient protection legislation. This research illustrates that the conventional wisdom regarding inferior care in HMOs may color how people assess their health care in surveys, resulting in survey findings biased toward showing HMOs provide inferior care and reinforcing existing stereotypes. Using merged data from the Community Tracking Study Household and Insurance Followback surveys, we identify privately insured persons who correctly and incorrectly know what kind of health plan they are covered by. Nearly a quarter misidentified their type of health coverage. Differences between responses by HMO and non-HMO enrollees to questions covering satisfaction with health care and physician choice, the quality of the last physician's visit, and patient trust in their physician shrink or disappear when we control for beliefs about what type of plan they are covered by. Results suggest that researchers and policy makers should be cautious about using consumer surveys to assess the relative quality of care provided under different types of health insurance. PMID- 12092674 TI - Health care report cards: implications for vulnerable patient groups and the organizations providing them care. AB - Standardized public reporting on the quality of health care (report cards) offers an opportunity to empower purchasers and consumers so that they can make choices that can result in better health care for less money. However, not all population subgroups are equally well served by the publication of such data. In particular, vulnerable patient groups such as the poor, the less educated, the chronically sick, and members of ethnic or linguistic minorities may find issues of importance to them largely neglected. In addition, the way that report card data are collected, analyzed, and presented may further marginalize the experiences of these groups who in any case are already underserved by the health system. This observation also has important implications for health care providers who serve primarily large numbers of vulnerable patients. The differential impacts of report card data on vulnerable patient groups (and their providers) need to be addressed by researchers and policy makers if access issues are not to be damaged further by the providers' pursuit of quality and value. PMID- 12092675 TI - Efforts to improve patient safety in large, capitated medical groups: description and conceptual model. AB - Medical care should be safer. Inpatient problems and solutions have received the most attention; this outpatient qualitative case study addresses a gap in knowledge. We describe safety improvements among large physician groups, model the key influences on their behavior, and identify beneficial public and private policies. All groups were trying to reduce medical injury, which was part of the sample design. The most commonly targeted problems are those that are similar across groups: shortcomings in diagnosis, abnormal tests follow-up, scope of practice and referral patterns, and continuity of care. Medical group innovators vary greatly, however, in implementation of improvements, that is, in the extent to which they implement process changes that identify events/problems, analyze and track incidents, decide how to change clinical and administrative practices, and monitor impacts of the changes. Our conceptual model identifies key determinants: (1) demand for safety comes from external factors: legal, market, and professional; (2) organizational responses depend on internal factors: group size, scope, and integration; leadership and governance; professional culture; information-system assets; and financial and intellectual capital. Further, safety is an aspect of quality (the same tools, decision making, interventions, and monitoring apply), and safety management benefits from prior efficiency management (similar skills and culture of innovation). Observed variation in even simple safeguards shows that existing safety incentives are too weak. Our model suggests that the biggest improvement would come from boosting the demand for quality and safety from both private and public larger group purchasers. Current policy relies too much on litigation and discipline, which have sometimes helped, but not solved, problems because they are inefficient, tend to drive needed information underground, and complicate needed cultural change. Patients' safety demand is also weak for want of information and market power. Big purchasers' demands, however, quickly influence the internal environment of medical groups, helping managers advance quality safety toward the top of groups' congested decision-making "queues." PMID- 12092676 TI - The use of expensive health technologies in the era of managed care: the remarkable case of neonatal intensive care. AB - The use of neonatal intensive care (NIC) continued to rise rapidly in the 1990s despite the concerns of observers about its cost effectiveness and its successes being mostly in facilities with high volume and capabilities. The objective of this study is to test the effects of insurance type, competition among hospitals, and market pressure from managed care plans on the supply and cost of NIC. The analysis uses logistic and linear models with techniques to avoid bias from (a) market area definitions based on actual patient flows and (b) self-selection of hospitals by patients with unmeasured risk of needing NIC. The data source contains all births in short-term hospitals in New Jersey during 1990 and 1994. Both the number of days and charges for NIC are reported. Key findings are that the decision of a hospital to offer NIC was associated with teaching status, the proportion of infants in the market area with documented high risk, and the market concentration of major competitors. The market share of managed care plans and the concentration of enrollment were not associated with either NIC being offered or with the standardized charges. Whether a particular patient was given to a NIC depended on patient risk factors and whether a NIC unit was present, but not on payer group. The results are consistent with the hypothesis that young insured parents (with the advice of their obstetricians) prefer hospitals with NIC and also are relatively profitable enrollees for health plans. IN CONCLUSION: using the results here and in other research, public and private policy makers may consider several ways to strengthen the incentives for health plans to contract for cost-effective birth-related services. The results also raise questions for a number of regulatory and payment policies and call for better public data on costs and outcomes for NIC. PMID- 12092677 TI - Policing Medicaid and Medicare managed care: the role of courts and administrative agencies. AB - The much-publicized 2000 case of Frew v. Gilbert, in which a federal judge castigated the State of Texas for deficiencies in its Medicaid program, brought renewed attention to the issue of regulating the quality of care in Medicaid and Medicare HMOs. Frew and other recent cases highlight both the promise and the pitfalls of relying on courts to correct deficiencies in public managed care programs. This article argues that while litigation over inadequacies in Medicare and Medicaid managed care can serve an important signaling function in alerting agencies and legislatures about the need for reform, the role of the courts in policing public managed care is circumscribed by several constraints. Barriers to class action litigation and differences in the institutional capacities of courts and administrative agencies mean that litigation is best viewed as a supplement, not an alternative, to a renewed commitment to strong quality monitoring on the part of Centers for Medicare and Medicaid Services and state Medicaid agencies. PMID- 12092678 TI - As good as it gets? The future of Medicare+Choice. PMID- 12092679 TI - Childhood brain tumors. AB - Primary malignant tumors of the central nervous system (CNS) account for about 16% of all childhood malignancies. These tumors are the second most common type of childhood cancer and the most frequent of the solid tumors. The small increase in incidence noted over the past two decades most likely represents advancements in diagnostic technologies rather than true changes in disease frequency, though this is controversial. CNS tumors are diverse, representing many histological types and arising in a variety of anatomic sites. The most common malignant tumors include astrocytomas (52%), medulloblastomas/primitive neuroectodermal tumors (PNETs) (21%), gliomas (19%), and ependymomas (9%). The current 5-year survival rate for all pediatric CNS tumors is 67%, but rates differ considerably among tumor types. Treatment modalities also differ according to histological type. Currently, about 25% of patients are treated with surgery alone, 40% undergo surgery plus radiation, and 30% are treated with surgery, radiation, and chemotherapy. Survivors of childhood brain tumors are at substantial risk for increased morbidity and late mortality. Five-year survivors of brain tumors are 13 times more likely to die than healthy age- and sex-matched peers. Disease recurrence remains the single most common cause of late deaths (70%). Neurological, neurocognitive, and endocrine disturbances are the most prevalent disabilities observed among long-term survivors of pediatric brain tumors. PMID- 12092680 TI - Use of magnetic resonance imaging in short stature: data from National Cooperative Growth Study (NCGS) Substudy 8. AB - The primary use of magnetic resonance imaging (MRI) in the evaluation of children with short stature (SS) is to discover lesions in the central nervous system (CNS), particularly tumors that may require intervention. MRI has a secondary role in identifying structural abnormalities responsible for growth hormone deficiency (GHD). We examined data from the National Cooperative Growth Study (NCGS) Substudy 8 to determine how American physicians are using MRI in evaluating children with SS. Of the 21,738 short children enrolled in NCGS, 5% underwent MRI during their follow-up. Children who had GH stimulation testing were more likely to have had an MRI than those in whom no GH stimulation test was performed (19% vs 2%, p <0.0001). Moreover, children diagnosed with severe GHD (maximum GH <5 ng/ml) were more likely to have an abnormal finding on MRI. Of these patients, 27% demonstrated an abnormality as compared to 12% and 12.5% in patients with partial GHD and normal GH stimulation test results (>10 ng/ml), respectively. Abnormalities unrelated to the hypothalamus or pituitary represented 30% of these findings, while disorders in pituitary anatomy, including pituitary hypoplasia, pituitary stalk interruption, and ectopic posterior pituitary, represented an additional 30% of abnormal MRI examinations. CNS tumors comprised 23% of abnormal findings in these patients. We conclude that MRI provides significant value in the evaluation of children with SS, by identifying CNS tumors associated with growth failure as well as anatomical abnormalities of the pituitary. These findings are useful in confirming the diagnosis of GHD in children and identifying potential candidates for continued GH replacement in adulthood. PMID- 12092681 TI - Utility of magnetic resonance imaging in the evaluation of the child with central diabetes insipidus. AB - Because of the association of central diabetes insipidus (CDI) and occult neoplasia, magnetic resonance imaging (MRI) is an important component of the diagnostic evaluation of a child with CDI. In more than 90% of these children, MRI (T1 weighted-image, without contrast) demonstrates an absence of the normal hyperintensity of the posterior pituitary. In one third of patients, the pituitary stalk is also thickened, suggesting infiltrative disease. Of those with a thickened stalk, the etiology of the CDI remains undetermined in about 60% of patients, whereas histiocytosis and occult germinoma each account for approximately 15-20% of patients. In contrast, germinoma is infrequent (3%) in children with CDI and an MRI showing a normal infundibular stalk, though histiocytosis still accounts for 15-20% of patients. In this paper, a diagnostic approach in children with CDI is proposed. PMID- 12092682 TI - Characteristics of growth hormone therapy for pediatric patients with brain tumors in the National Cooperative Growth Study (NCGS) and from a survey of pediatric endocrinologists. AB - Pediatric patients with brain tumors may have significant short stature secondary to growth hormone deficiency (GHD). This occurs as a result of impaired hypothalamic function due to tumor location, radiation therapy, surgery, or chemotherapy. The use of GH replacement therapy in this patient population is variable and somewhat unpredictable. We analyzed patient data from the National Co-operative Growth Study (NCGS) database and a survey completed by 19 members of the Pediatric Endocrine Alliance for Neuro Oncology Patients (PEANOP) to identify patterns of GH use in pediatric patients following treatment for brain tumors. From the NCGS database, we present demographic, dosing, and safety data. The PEANOP survey was examined to determine the percentage of patients receiving GH and the likelihood of treating these patients with GH following tumor treatment. The time to initiating GH replacement therapy was evaluated in 14 tumor types, as well as the contributing impact of the extent of tumor resection. PMID- 12092683 TI - Septo-optic dysplasia/optic nerve hypoplasia: data from the National Cooperative Growth Study (NCGS). AB - We analyzed data from 65 children with septo-optic dysplasia (SOD) referred for evaluation and followed in the National Cooperative Growth Study (NCGS) Substudy 8 and from 758 children treated with growth hormone (GH) and followed in the NCGS core study. Compared to other children referred for evaluation of short stature, children with SOD were younger (mean age 3.7 +/- 3.6 vs 8.6 +/- 4.9 years), had less severe short stature (mean +/- SD height SDS -1.80 +/- 1.64 vs -2.17 +/- 0.95), and were more likely to be female (46% F vs 31% M). Children with SOD who received GH were older and shorter than those referred and untreated, but the gender distribution was similar. Other pituitary hormone deficits were reported in untreated patients, including thyroid hormone deficiencies (8%) and adrenocorticotropic hormone (ACTH) deficiency (3%), as compared to 27% and 24%, respectively, in GH-treated children. Data on adult height were available for 71 patients, who showed an average gain in height SDS of 1.17 +/- 1.49. GH therapy was well tolerated in children with SOD. PMID- 12092684 TI - Genetic disorders of human growth. AB - Growth in humans is a complex process, controlled at numerous levels and by a myriad of factors. These factors may act centrally or peripherally and may be hormones, receptors, or transcription factors. Many of these probably are still unknown. The factors that are discussed here include those that act on the developing pituitary gland (transcription factors including LHX3, HESX1, PROP1, and PIT-1); those that regulate the normal activity of the pituitary (hypothalamic hormones such as GHRH, Ghrelin [growth hormone secretagoguel and somatostatin); those factors coming from the pituitary (essentially growth hormone [GH]); and the downstream modulators, transducers, and effectors of GH (including the GH receptor/GH binding protein, insulin-like growth factor-I and II, their receptors, and their binding factors). What is becoming increasingly clear is the role of genetics in determining stature. This review discusses the most clinically relevant factors, with an emphasis on ontogeny, genetic inheritance, and clinical presentation. PMID- 12092685 TI - A long-acting human growth hormone (Nutropin Depot): efficacy and safety following two years of treatment in children with growth hormone deficiency. AB - BACKGROUND: Nutropin Depots [somatropin (rDNA origin) for injectable suspension] is a long-acting form of human growth hormone (GH) to be administered by subcutaneous (s.c.) injection. The availability of this formulation offers the opportunity for greater convenience and compliance by decreasing the number of injections and frequency of administration required. OBJECTIVE: To determine the efficacy and safety of a long-acting formulation of GH administered in children with GH deficiency (GHD) once or twice monthly for 2 years. PATIENTS: Fifty-six previously untreated, prepubertal children with GHD received Nutropin Depot 1.5 mg/kg once monthly (1x/mo), or 0.75 mg/kg twice monthly (2x/mo) for 24 months. The mean pretreatment growth rate was 5.0 +/- 2.4 cm/yr. RESULTS: The 0-12 mo growth rate (mean +/- SD) was 8.3 +/- 1.5 cm/yr in the 1x/mo group and 8.2 +/- 2.0 cm/yr in the 2x/mo group. The 12-24 month growth rate was 7.2 +/- 1.5 cm/yr in the 1x/mo group and 6.9 +/- 1.5 cm/yr in the 2x/mo group. During the 24 months of treatment, height standard deviation score (SDS) increased by 1.0 +/- 0.5 SD in the two groups combined (p <0.0001). The corresponding advancement in bone age was 2.2 +/- 0.7 yr, resulting in a gain in Bayley-Pinneau predicted adult height (PAH) SDS of 0.6 +/- 0.9 SD in the 1x/mo group and 0.6 +/- 1.0 SD in the 2x/mo group. No serious adverse events attributable to the study drug were reported. Injection site reactions were common, but resolved without intervention. Pre-dose fasting and postprandial glucose and insulin levels, as well as hemoglobin A1c levels, were unchanged from baseline values. CONCLUSIONS: Treatment with Nutropin Depot is associated with catch-up growth and normal skeletal maturation and is a viable alternative to daily injections of GH in children with GHD. PMID- 12092686 TI - Growth hormone treatment and pseudotumor cerebri: coincidence or close relationship? AB - Pseudotumor cerebri (PTC) is an uncommon disorder in the pediatric population. It is not a benign condition. It can cause permanent vision loss. The most recently recognized risk factor for this disorder is recombinant human growth hormone (GH) therapy. Data from Genentech's National Cooperative Growth Study (NCGS), a postmarketing surveillance program, are analyzed to examine the relationship between GH therapy and PTC. Several areas are addressed, including plausibility, probability, clinical and laboratory presentations, management, clinical outcome, present state of knowledge, and future recommendations. PMID- 12092687 TI - Growth problems and growth hormone treatment in children with cystic fibrosis. AB - Children with cystic fibrosis (CF) typically demonstrate poor linear growth and, despite aggressive nutritional supplementation, suboptimal weight gain. Growth velocity may be particularly blunted during puberty, a finding that occurs independently of clinical status or sex steroid levels. Several studies indicate that growth hormone (GH) treatment may be associated with significant improvement in both height velocity and weight gain in children with CF. These findings may translate into improvements in clinical status. In our experience, improved growth and weight status have been associated with fewer hospitalizations, fewer antibiotic courses, and improved pulmonary function. A large multicenter trial is currently underway to assess the effect of long-term therapy with GH on the quality of life and pulmonary status. PMID- 12092688 TI - Type 2 diabetes mellitus in children and youth: a new epidemic. AB - Type 2 diabetes mellitus (DM) has been described as a new epidemic affecting the American pediatric population. This is coincident with an overall 33% increase in DM prevalence documented during the last decade. In 1992, type 2 DM was a rare occurrence in most pediatric centers. By 1994, it represented up to 16% of new cases in urban areas, and by 1999, the incidence of new type 2 DM diagnoses ranged between 8% and 45%, depending on geographic location. These patients have been observed primarily in African American, Mexican American, Native American, and Asian American children and youth. As in the adult population, type 2 DM in children and youth occurs as a result of insulin resistance coupled with relative beta-cell failure. While there appears to be a host of potential genetic and environmental risk factors for these aberrations, perhaps the most significant risk factor is obesity. Other risk factors include a family history of type 2 DM, puberty, intrauterine exposure to DM, sedentary lifestyle, female gender, and certain ethnicities. To date, few studies have addressed the role of physical activity and nutrition counseling in improving glycemic outcome, the most effective ways to reduce cardiovascular risk, or the most effective treatment regimens for this population. Once type 2 DM is established, the persistence of obesity often interferes with the response to treatment and exacerbates the comorbidities of hypertension, dyslipidemia, atherosclerosis, and polycystic ovarian syndrome (PCOS). Since fewer than 10% of youth with type 2 DM can be treated with diet and exercise alone, pharmacological intervention is generally required to achieve normoglycemic targets. In most surveys, practitioners prescribe insulin or an oral agent, most often metformin. Specific treatment algorithms for pediatric patients with type 2 DM need to be rigorously investigated. PMID- 12092690 TI - Exercise, diet, and childhood obesity: the GH-IGF-I connection. AB - The recent "obesity epidemic" among children and adolescents is a major public health concern. The mechanisms responsible for the increased incidence of childhood obesity are not yet well understood. The absence of a clear mechanism makes treating the obese child or adolescent a difficult task, and standardized therapeutic approaches simply do not yet exist. Metabolic derangements associated with obesity may contribute to the difficulty in treatment. Observed abnormalities in the growth hormone (GH)-insulin-like growth factor-I (IGF-I) axis in obese adults and the impact of exercise on the GH-IGF-I system are of particular relevance to the growing obese child. In this review, we focus on the interacting mechanisms of diet and exercise through specific hormonal mediators and their contribution to the current obesity epidemic. An improved understanding of these mechanisms may be helpful in creating effective treatment programs for children with obesity. PMID- 12092689 TI - Progress in the treatment of childhood diabetes mellitus and obesity. AB - The 61st Annual Meeting and Scientific Sessions of the American Diabetes Association (ADA) in Philadelphia, PA, (June 22-26, 2001) presented many topics of interest to pediatric clinicians. Of particular interest were the results of the insulin injection arm of the Diabetes Prevention Trial for type 1 diabetes mellitus (DM) (DPT-1). Over 80,000 relatives of patients with type 1 DM were screened. Ultimately, 339 subjects were randomized either to active therapy (twice daily insulin injections plus an annual insulin infusion) or to close observation. Risk prediction algorithms appeared to be accurate. Unfortunately, however, insulin therapy did not decrease the risk of developing DM. Of note, this was primarily a pediatric study with most of those randomized under 21 years of age. As expected, young subjects (<12 years) progressed toward the development of DM at a faster rate than older subjects (>15 years). The second arm of the DPT 1 trial, testing oral insulin in those with intermediate risk (25-50%) for DM, is still recruiting subjects. The controversial topic of continuous subcutaneous insulin infusion (CSII) in young children was also addressed. Many investigators presented data strongly supporting the successful use of infusion pumps in young children. In general, glycemic control was improved or remained stable, the incidence of severe hypoglycemia was low, and families reported more flexibility in their lifestyle. Obesity, an increasing problem in pediatric patients, was also addressed. PMID- 12092691 TI - Planning medical services for the elderly: disability free survival must be the goal. PMID- 12092692 TI - Cost effectiveness of statins for the secondary prevention of coronary heart disease in Ireland. PMID- 12092693 TI - Tuberculosis in the Republic of Ireland at the end of the 1990s. AB - There has been limited detailed epidemiological data available on tuberculosis in the Republic of Ireland. The 1998 and 1999 National TB Reports produced by the National Disease Surveillance Centre presented disaggregate national data describing in detail the epidemiology of TB in the Republic of Ireland. Individual case notifications were collated by health boards, forwarded to NDSC where they were entered onto a national TB database and then analysed using Epi Info. There were 893 cases of TB notified in 1998-1999. It was more common in older age groups and men. 50% of cases occurred in those less than 45 years, an indication of considerable ongoing transmission of tuberculosis. Regional variation in the rate of TB exists and a relatively small proportion of cases occurred in foreign-born patients. TB in HIV positive patients was not common and MDR-TB has also been observed. TB has not disappeared from the Republic of Ireland. Treatment, contact tracing and surveillance need to be maintained and preferably enhanced. PMID- 12092694 TI - A prospective study for evaluation of knee effusion after hip surgery. AB - Knee effusion is a common occurrence in elderly patients following hip surgery. We prospectively studied 106 patients undergoing proximal femoral surgery. These patients were examined pre-operatively, on days 4 and 8, and six weeks post operatively for the presence of an ipsilateral knee effusion. 71 patients had undergone Total Hip Arthroplasty (THA), 26 Dynamic Hip Screw (DHS) insertion and 9 had hemiarthroplasty. The incidence of pre-operative ipsilateral knee effusion was 13% (14 patients). 42% (44 patients) had an effusion at one week post operatively and 12% (13 patients) at 6 weeks. 57% (60 patients) of the study group had radiological evidence of osteoarthritis in their knees. However, the incidence of effusion in patients with and without arthritis was the same. The majority of these effusions resolved after six weeks with no residual effects. PMID- 12092695 TI - Referral to physiotherapy as part of a general practitioner's management of back pain. AB - The aim of this survey was to investigate how general practitioners (GPs) manage their back pain patients and if they refer them for physiotherapy. The research instrument was a self-administered postal questionnaire. This was sent to 580 GPs, one quarter of the GPs in the Republic of Ireland. Two hundred and ninety three questionnaires (n=293)were returned, which constituted a 50.5% response rate. Two hundred and sixty one questionnaires (n=261) were usable. The main findings of the study were that the respondents consider that physiotherapy has a major role in the management of back pain. However there was limited availability of physiotherapy for the GPs' public patients. One hundred and thirteen GPs (over 43% of respondents) have referred back pain patients to alternative practitioners for manipulation. One hundred and forty four respondents (52%) believe that physiotherapists should not treat back pain patients without the patient first consulting their doctor. PMID- 12092696 TI - The use of local accident and emergency injury surveillance to monitor the impact of a lay safety community programme. AB - To study prospectively the injury patterns in under 14 year olds presenting to casualty. To use this information to assess the impact of a local Community Childhood Accident Prevention Project (CCAPP) Prospective injury surveillance was collected on all attendees under 14 years of age. Casualty attendance for the members of the Safety Club and matched controls were analysed. Of the 4,267 attendees there was the expected male predominance. 2,261 (53%) of injuries occurred at home, 574 (13.5%) on the road, 553 (13%) at school with 202 (4.7%) during sports. Priority areas noted were high falls > 1 metre, road accidents, burns and poisonings. Those participating in the program demonstrated significantly (p-value < 0.05) reduced admission rates and head injury rates when compared to non-participants. The above database allows us to analyse injury patterns and to assess the success of an EU-sponsored local injury prevention campaign in areas of high social deprivation. PMID- 12092697 TI - Acute vascular events associated with cisplatin therapy in malignant disease. PMID- 12092698 TI - Multiple myeloma in young persons. PMID- 12092700 TI - Is the EU working time directive making teamwork obsolete. PMID- 12092699 TI - A misleading patient in intensive care. PMID- 12092701 TI - The Mental Health Act 2001. PMID- 12092702 TI - The Mental Health Act, 2001: a brief summary. PMID- 12092703 TI - Lessons from America: trials and tribulations of a specialist registrar. PMID- 12092704 TI - Nail gun injuries. PMID- 12092705 TI - Pathogenesis and pathology of hematogenous infections of the fetus and newborn. AB - The earlier infections occur in intrauterine life the more severe are they. When the infection develops during embryogenesis, the lesions are much more serious, sometimes causing disruptions (malformations). At this time the organs are not completely formed and microorganisms may interfere with organogenesis to such an extent that the development of the functions necessary for viability become impaired. Infection acquired in utero may result in resorption of the embryo, abortion, stillbirth, neonatal death, intrauterine growth retardation (IUGR), or prematurity. The infected newborns commonly are apparently normal at birth but they may develop a late onset disease. Otherwise, neonates presenting symptomatology may derelop untoward sequelae. PMID- 12092707 TI - Comparison of immortalization assay and polymerase chain reaction detection of Epstein-Barr virus in pediatric transplant recipients and control samples. AB - The polymerase chain reaction technique (PCR) has become a standard for the detection of Epstein-Barr virus ( EBV). Several studies have shown that solid organ transplant recipients fated to develop post-transplant lymphoproliferative disease (PTLD) have higher mean levels of EB viremia as measured by PCR. We directly compared PCR and immortalization assay (IA) for EBV detection from control cell lines as well as from peripheral blood monouclear cells (PBMC) and saliva samples from 9 pediatric solid organ transplant recipients. Results were expressed as the lowest amount of DNA and/or number of cells detected. All samples negative for EBV by PCR had amplifiable DNA detected using beta-actin primers. When control cell lines were assayed and positive in both assays, PCR was at least 200 times more sensitive than IA, as expected. When PBMC and saliva were compared, 1 patient, with a mononucleosis-like syndrome, was positive for EBV by IA only. When both IA and PCR were positive however, PCR was able to detect 5-500 times less EBV than IA, as expected. The associations of positive test with each assay and the diagnosis of PTLD were similar. PMID- 12092706 TI - The placenta in hematogenous infections. AB - Although congenital infections have been known since ancient times, at present they constitute an evolution due to the broadening of diagnostic methods. Also, the histopathological examination of selected placentas has increased in perinatal pathology. All placentas should be examined grossly by the obstetrician or the neonatologist who must select the specimens to be sent for pathologic examination. The indications for selection at delivery to detect hematogenous infection are listed in Table 1. The placentas that do not meet guidelines for pathologic examination must be kept stored at 40 degrees C for at least 72 h. This routine ensures that the placentas of asymptomatic newborns who might present early manifestations suggestive of infection would be submittedfor pathologic examination. PMID- 12092708 TI - Pathology teach and tell: large cell lymphoma--T-cell type. PMID- 12092709 TI - Temporal and spatial concepts in child language: conventional and configurational. AB - The purpose of this research was to study the temporal and spatial systems of child language. Configurational time/space refers to the location of events/objects relative to referent events/objects (e.g., before/after or across/along) and conventional time/space refers to the specification of cycles/directions and the measurement of duration/distance (e.g., winter/fall or North/South). The research was designed to determine how configurational and conventional notions of time and space emerge in the child's language. The average age of the children in the cross-sectional design was: 3;9, 6:7, and 8:7. The children received a comprehension test that required them to match a sentence with a picture. In general, configurational contrasts were easier to comprehend than conventional contrasts, and, within the configurational problems, location requiring a single referent was easier than location requiring multiple referents. There was no overall difference between temporal and spatial dimensions. The role of conceptual development was discussed within the context of a comparison between first and second language acquisition. PMID- 12092710 TI - Word order in sentence processing: an experimental study of verb placement in German. AB - We examine the question of whether the human comprehension device exhibits word order preferences during on-line sentence comprehension. The focus is on the positioning of finite verbs and auxiliaries relative to subjects and objects in German. Results from three experiments (using self-paced reading and event related brain potentials) show that native speakers of German prefer to process finite verbs in second position (i.e., immediately after the subject and before the object). We will account for this order preference in terms of the relative processing costs associated with SVfO and SOVf. Our finding that word-order preferences play an important role in the on-line comprehension of German sentences is compatible with results from previous studies on English and other languages. PMID- 12092711 TI - "If you and I, if we, in this later day, lose that sacred fire . . .": perspective in political interviews. AB - It is argued in the following that the dialogical complexity of speaker perspective requires a broad empirical analysis. To date, such analyses, particularly of political discourse, have been couched in terms of narrower concepts, such as self-presentation and political positioning or involvement/distancing, and have been typically carried out by means of qualitative methods applied to pronominal usage. The present research applies complementarily both quantitative and qualitative analyses to BBC television interviews of Shimon Peres (January 29, 2001) and of Edward Said (October 18, 2000) by Tim Sebastian in a program entitled HARDtalk. In addition to pronouns, these analyses include a number of other hypothetical indicators of a broad concept of perspective on the part of both interviewer and interviewees: turn initial words, hesitations, questions, use of yes and no, personal reference utterances (e.g., I think), interjections, number of syllables spoken, and interruptions and overlaps. Quantitative comparisons of interviewer with interviewee revealed important differences on all these measures. Qualitative analyses also confirmed subtle local dynamics of perspective. Accordingly, the findings are interpreted within a general theoretical concept of perspective, derived from Bakhtin's (1981) dialogicity. PMID- 12092712 TI - Orthographic neighbors and visual word recognition. AB - Two lexical decision experiments, using words that were selected and closely matched on several criteria associated with lexical access provide evidence of facilitatory effects of orthographic neighborhood size and no significant evidence of inhibitory effects of orthographic neighborhood frequency on lexical access. The words used in Experiment 1 had few neighbors that were higher in frequency. In Experiment 2, the words employed had several neighbors that were higher in frequency. Both experiments showed that words possessing few neighbors evoked slower responses than those possessing many neighbors. Also, in both experiments, neighborhood size effects occurred even though words from large neighborhoods had more potentially interfering higher-frequency neighbors than words from small neighborhoods. PMID- 12092713 TI - Serotyping and ribotyping of Salmonella using restriction enzyme PvuII. AB - The subtyping and identification of bacterial pathogens throughout food processing and production chains is useful to the new hazard analysis critical control point-based food safety plans. Traditional manual serotyping remains the primary means of subtyping Salmonella isolates. Molecular biology techniques, however, offer the promise of more rapid and sensitive subtyping of Salmonella. This study evaluates the potential of restriction enzyme PvuII, followed by probing with the rRNA operon from Escherichia coli, to generate serotype-specific DNA fingerprints. A total of 32 identified serotypes were found with an overall agreement in 208 of the 259 (80%) isolates tested between U.S. Department of Agriculture serotype identification and riboprint serotype identification. Many of the isolates that did not correlate were serotype identified as Salmonella Montevideo, which indicates that for this serotype, there are multiple ribotypes. When Salmonella Montevideo isolates were not included, the ribotype identification agreed with serotyping in 207 of the 231 (90%) isolates. The primary outcome of any ribotyping procedure is to give distinct ribotype patterns. This extensive poultry epidemiological study demonstrates that, in addition to ribotype patterns, the identification of isolates to known serotypes provides the investigator with additional information that can be more useful than traditional epidemiology and isolate identification studies. PMID- 12092714 TI - Humectant permeability influences growth and compatible solute uptake by Staphylococcus aureus subjected to osmotic stress. AB - The effects of different humectants (sodium chloride, sucrose, and glycerol) on the growth of and compatible solute (glycine betaine, proline, and carnitine) uptake by the osmotolerant foodborne pathogen Staphylococcus aureus were investigated. While growth in the presence of the impermeant humectants sodium chloride and sucrose induced the accumulation of proline and glycine betaine by cells, growth in the presence of the permeant humectant glycerol did not. When compatible solutes were omitted from low-water-activity media, growth was very poor in the presence of impermeant humectants. In contrast, the addition of compatible solutes had essentially no effect on growth when cells were grown in low-water-activity media containing glycerol as the humectant. Carnitine was found to accumulate to high intracellular levels in osmotically stressed cells when proline and glycine betaine were absent, making it a potentially important compatible solute for this organism. PMID- 12092715 TI - Molecular identification of nine commercial flaffish species by polymerase chain reaction-restriction fragment length polymorphism analysis of a segment of the cytochrome b region. AB - Commercial refrigerated or frozen flatfish fillets are sometimes mislabeled, and identification of these mislabeled products is necessary to prevent fraudulent substitution. Identification of nine commercial flatfish species (order Pleuronectiformes), Hippoglossus hippoglossus (halibut), Lepidorhombus boscii (four-spotted scaldfish), Lepidorhombus whiffiagonis (megrin), Platichthys flesus (flounder), Pleuronectes platessa (European plaice), Reinhardtius hippoglossoides (Greenland halibut), Scophthalmus maximus (turbot), Scophthalmus rhombus (brill), and Solea vulgaris (=Solea solea) (sole), was carried out on the basis of the amplification of a 486-bp segment of the mitochondrial genome (tRNA(Glu)/cytochrome b) by using the polymerase chain reaction (PCR) and universal primers. Sequences of PCR-amplified DNA from the flatfish species were used to select eight restriction enzymes (REs). The PCR products were cut with each RE, resulting in species-specific restriction fragment length polymorphism. Seven species groups could be identified by application of the single RE DdeI and six species groups by using HaeIII, HinfI, MaeI, or MboI. Different combinations of only a couple of these REs could unambiguously identify the nine flatfish species. Genetic polymorphisms of the target sequence were examined by comparison with previously published DNA sequences, and the results of this comparison confirmed the usefulness of this technique in distinguishing and genetically characterizing refrigerated or frozen pieces of these nine flatfish species. PMID- 12092716 TI - Influence of chilling methods on the quality of sardines (Sardina pilchardus). AB - The aim of this study was to determine for sardines (Sardina pilchardus) the effect of (i) chilling in ice and water in small expanded polystyrene boxes during distribution and in ice thereafter and (ii) chilling in ice and water for the entire storage period. These storage methods were compared with storage of the fish in ice in wooden boxes or in expanded polystyrene boxes. Three storage experiments were performed to take into account the variability in handling conditions and seasonality. There were significant (P < 0.05) differences in the results of some of the sensory, physical, chemical, and microbiological analyses performed that showed that the fish were of better quality when preserved in water and ice than when preserved in ice alone. The effects of dewatering of the fish after transport and subsequent storage in ice were also significantly different (P < 0.05) from those of storage in ice, with the former storage method resulting in better sensory and microbial indices. However, oxidative rancidity, as measured by thiobarbituric acid-reactive substances, was higher for the dewatered fish than for fish preserved in water and ice throughout the storage period. It is concluded that for sardines, storage in water and ice, either only during transport or throughout the storage period, can be useful as an alternative preservation method during distribution and retail sale. PMID- 12092717 TI - Survey of hydrogen sulphide production by wine yeasts. AB - Twenty-one strains of commercial wine yeasts and 17 non-Saccharomyces species of different provenance were surveyed for their ability to produce hydrogen sulphide in synthetic grape juice medium indicator agar with different nitrogen sources, as well as in natural grape juice. Bacto Biggy agar, a commercially available bismuth-containing agar, was used to compare our results with others previously reported in the literature. Under identical physiological conditions, the strains used in this study displayed similar growth patterns but varied in colony color intensity in all media, suggesting significant differences in sulphite reductase activity. Sulphite reductase activity was absent for only one strain of Saccharomyces cerevisiae. All other strains produced an off-odor to different extents, depending significantly (P <0.05) on medium composition. Within the same species of some non-Saccharomyces yeasts, strain variation existed as it did for Saccharomyces. In natural musts, strains fell into three major groups: (i) nonproducers, (ii) must-composition-dependent producers, and (iii) invariable producers. In synthetic media, the formation of sulphide by strains of S. cerevisiae results from the reduction of sulphate. Therefore, this rapid screening methodology promises to be a very useful tool for winemakers for determining the risk of hydrogen sulphide formation by a given yeast strain in a specific grape juice. PMID- 12092718 TI - Acquisition of antibiotic resistance plasmids by enterohemorrhagic Escherichia coli O157:H7 within rumen fluid. AB - The emergence of antibiotic resistance among important foodborne pathogens like Escherichia coli O157:H7 has become an important issue with regard to food safety. In contrast to the case for Salmonella, antibiotic resistance has been slow in its development in E. coli O157:H7 despite the presence of mobile antibiotic resistance genes in other E. coli organisms that inhabit the same animal host. We set out to determine if rumen fluid influences the transfer of plasmid-mediated, antibiotic resistance to E. coli O157:H7. A commensal E. coli strain from a dairy cow was transformed with conjugative R plasmids and served as the donor in matings with naladixic acid-resistant E. coli O157:H7. R plasmids were transferred from the donor E. coli strain to E. coli O157:H7 in both Luria Bertani (LB) broth and rumen fluid. R plasmids were transferred at a higher frequency to E. coli O157:H7 during 6 h of incubation in rumen fluid at rates comparable to those in LB broth, indicating that conditions in rumen fluid favor the transfer of the plasmids to E. coli O157. This finding suggests that the cow's rumen is a favorable environment for the genetic exchange of plasmids between microflora and resident E. coli O157:H7 in the bovine host. PMID- 12092719 TI - Salmonella serotypes isolated from nonhuman sources in Sao Paulo, Brazil, from 1996 through 2000. AB - A total of 4,581 Salmonella strains isolated from nonhuman sources, including foodstuffs associated with foodborne Salmonella outbreaks, from January 1996 through December 2000 were serotyped at the Enteropathogens Laboratory, Instituto Adolfo Lutz, Sao Paulo, Brazil. Among the 123 different serotypes identified, Salmonella enterica subsp. enterica serotype Enteritidis (Salmonella Enteritidis) was the most prevalent (32.7%), ranking first for almost every kind of source. The next most common serotypes were Salmonella Senftenberg (10.3%), Salmonella Hadar (6.8%), Salmonella Agona (5.1%), and Salmonella Typhimurium (2.4%). Rough strains belonging to the subspecies S. enterica subsp. enterica (4.8%), S. enterica subsp. arizonae (<1%), S. enterica subsp. diarizonae (<1%), and S. enterica subsp. houtenae (<1%) were also detected. Foodstuffs (including poultry meat for consumption) contained 38.1% of the studied Salmonella strains, poultry flocks (from several farms under salmonellosis control by the owners) contained 21.7%, the environment contained 10.6%, sewage contained 9.4%, water contained 6.6%, animal feed contained 4.4%, chill water from poultry-processing operations contained 2.2%, and other sources contained 7.0%. Foodstuffs extensively contaminated with Salmonella strains were poultry meat (40%), cow meat (11%), desserts (8%), mayonnaise (6%), sausage (5%), and unpasteurized shell eggs (4%), and there were several other food sources (26%). Homemade mayonnaise was the most common vehicle for Salmonella foodborne outbreaks, and Salmonella Enteritidis was the serotype most isolated (95%) from that source. According to these data and previously published data concerning Salmonella strains isolated in Sao Paulo State, almost the same serotypes have predominated among nonhuman sources for the last decade. PMID- 12092720 TI - Effects of wearing knitted or rubber gloves on the transfer of Escherichia cohi between hands and meat. AB - On eight occasions, five volunteers each handled five pieces of meat with bare hands or while wearing dry or wet knitted gloves or rubber gloves after hands had been inoculated with Escherichia coli or after handling a piece of meat inoculated with E. coli. On each occasion, after all meat was handled, each piece of meat, glove, and hand were sampled to recover E. coli. When hands were inoculated, E. coli was recovered from all meat handled with bare hands, in lesser numbers from some pieces handled with knitted gloves, and from only one piece handled with rubber gloves. When pieces of inoculated meat were handled, the numbers of E. coli transferred to uninoculated meat from bare hands or rubber gloves decreased substantially with each successive piece of uninoculated meat, but decreases were small with knitted gloves. The findings indicate that, compared with bare hands, the use of knitted gloves could reduce the transfer of bacteria from hands to meat but could increase the transfer of bacteria between meat pieces, whereas the use of rubber gloves could largely prevent the first and greatly reduce the second type of bacteria transfer. PMID- 12092721 TI - A rapid and improved method for the detection of Vibrio parahaemolyticus and Vibrio vulnificus strains grown on hydrophobic grid membrane filters. AB - DNA probe-based detection methods were developed and characterized as an alternative to time-consuming and less specific conventional protocols. Digoxigenin-labeled probes were prepared by polymerase chain reaction amplification of the targeted sequences in the specific amplicons generated from genomic DNA. Specific probes with high yields were generated for the detection of the tlh gene of Vibrio parahaemolyticus and the cth gene of V. vulnificus. Colony (Southern) hybridization analyses were carried out using hydrophobic grid membrane filters (HGMFs) to allow biotype-specific differentiation of the two species. Eight strains of V. vulnificus and five strains of V. parahaemolyticus, including one standard (ATCC) strain of each biotype, were examined. Colony lysis, hybridization, and nonradioactive detection parameters were optimized for identification of the target biotypes arranged on the same HGMF and also on a conventional nylon membrane, thereby confirming the specificity of the probes and the comparative usefulness of the HGMFs. The experimental procedure presented here can be completed in 1 day. The protocol was designed specifically to identify the target Vibrio spp. and could potentially be used for the enumeration and differentiation of V. parahaemolyticus and V. vulnificus in foods. PMID- 12092722 TI - Efficacy of sanitation and cleaning methods in a small apple cider mill. AB - The efficacy of cleaning and sanitation in a small apple cider processing plant was evaluated by surface swab methods as well as microbiological examination of incoming raw ingredients and of the final product. Surface swabs revealed that hard-to-clean areas such as apple mills or tubing for pomace and juice transfer may continue to harbor contaminants even after cleaning and sanitation. Use of poor quality ingredients and poor sanitation led to an increase of approximately 2 logs in aerobic plate counts of the final product. Reuse of uncleaned press cloths contributed to increased microbiological counts in the finished juice. Finally, using apples inoculated with Escherichia coli K-12 in the plant resulted in an established population within the plant that was not removed during normal cleaning and sanitation. The data presented in this study suggest that current sanitary practices within a typical small cider facility are insufficient to remove potential pathogens. PMID- 12092723 TI - Development, implementation, and analysis of an on-farm food safety program for the production of greenhouse vegetables. AB - Fresh fruits and vegetables are increasingly recognized as vectors for foodborne illness. Consequently, an on-farm food safety program was developed, implemented, and analyzed for the Ontario Greenhouse Vegetable Growers in Ontario, Canada, during a 2 1/2-year period. This hazard analysis critical control point-based system was designed to reduce the potential of microbial contamination along the entire production and distribution process. Through the use of microbiological testing, on-site visits, and producer surveys, it was determined that the program has increased grower knowledge, understanding, and awareness of microbial risks associated with fresh produce and caused improvements in practices used within the greenhouse and packing sheds. PMID- 12092724 TI - Behavior of Listeria monocytogenes inoculated on cantaloupe surfaces and efficacy of washing treatments to reduce transfer from rind to fresh-cut pieces. AB - Attachment and survival of Listeria monocytogenes on external surfaces (rind) of inoculated cantaloupe, resistance of the surviving bacteria to chlorine or hydrogen peroxide treatments, transfer of the pathogen from unsanitized and sanitized rinds to fresh-cut tissues during cutting and growth, and survival of L. monocytogenes on fresh-cut pieces of cantaloupe were investigated. Surface treatment with 70% ethanol to reduce the native microflora on treated melon, followed by immersion in a four-strain cocktail of L monocytogenes (10(8) CFU/ml) for 10 min, deposited 4.2 log10 CFU/cm2 and 3.5 log10 CFU/cm2 of L monocytogenes on treated and untreated cantaloupe rinds, respectively. L. monocytogenes survived on the treated or untreated cantaloupe rinds for up to 15 days during storage at 4 and 20 degrees C, but populations declined by approximately 1 to 2 log10 CFU/cm2. Fresh-cut pieces prepared from inoculated whole cantaloupes stored at 4 degrees C for 24 h after inoculation were positive for L. monocytogenes. Washing inoculated whole cantaloupes in solutions containing 1,000 ppm of chlorine or 5% hydrogen peroxide for 2 min at 1 to 15 days of storage at 4 degrees C after inoculation resulted in a 2.0- to 3.5-log reduction in L. monocytogenes on the melon surface. Fresh-cut pieces prepared from the sanitized melons were negative for L. monocytogenes. After direct inoculation onto fresh cut pieces, L. monocytogenes survived, but did not grow, during 15 days of storage at 4 degrees C. Growth was evident by 4 h of storage at 8 and 20 degrees C. It is concluded that sanitizing with chlorine or hydrogen peroxide has the potential to reduce or eliminate the transfer of L. monocytogenes on melon surfaces to fresh-cut pieces during cutting. PMID- 12092725 TI - Potential for the spread of Escherichia coli O157, Salmonella, and Campylobacter in the lairage environment at abattoirs. AB - Prevalences of Escherichia coli O157, Salmonella spp., and Campylobacter spp. were examined in 270 swabs taken from selected sites along the unloading-to slaughter routes of animal movement in lairages of six commercial abattoirs, three for cattle and three for sheep. The overall prevalences of the pathogens in the respective lairage environments were compared with those for 270 swabs from the pelts of 90 lambs examined in the present study and 270 swabs from the hides of 90 cattle examined in a previous study that were slaughtered at the same abattoirs on the same days. Also, the results obtained were analyzed with the aim of identifying critical points at which animal-environment-animal transfer of the pathogens in lairages occurs. The results showed that (i) the overall prevalences of E. coli O157, Salmonella spp., and Campylobacter spp. were 27.2, 6.1, and 1.1%, respectively, in cattle lairages and 2.2, 1.1, and 5.6%, respectively, in sheep lairages; (ii) the overall prevalences of the three pathogens on cow hides (28.8, 17.7, and 0%, respectively) and sheep pelts (5.5, 7.8, and 0%, respectively) were higher than the overall prevalences in the respective lairage environments; (iii) the most frequently contaminated sites in cattle lairages were holding pen floors (50% of swabs positive for one or more pathogens), entrance gates of stun boxes (27.8% of swabs positive for one or more pathogens), and stun box floors (22.2% of swabs positive for one or more pathogens); (iv) the most frequently contaminated sites in sheep lairages were unloading ramp floors, holding pen floors, and water troughs (33.3, 22.2, and 22.2%, respectively); and (v) overall, cattle lairages and cow hides were more frequently contaminated with the pathogens than were lamb lairages and lamb pelts. Further research is needed to develop strategies for the incorporation of pathogen control in lairages into integrated microbial meat safety systems. PMID- 12092726 TI - Testing for Salmonella in raw meat and poultry products collected at federally inspected establishments in the United States, 1998 through 2000. AB - The Food Safety and Inspection Service (FSIS) issued Pathogen Reduction; Hazard Analysis and Critical Control Point (HACCP) Systems; Final Rule (the PR/HACCP rule) on 25 July 1996. To verify that industry PR/HACCP systems are effective in controlling the contamination of raw meat and poultry products with human disease causing bacteria, this rule sets product-specific Salmonella performance standards that must be met by slaughter establishments and establishments producing raw ground products. These performance standards are based on the prevalence of Salmonella as determined from the FSIS's nationwide microbial baseline studies and are expressed in terms of the maximum number of Salmonella positive samples that are allowed in a given sample set. From 26 January 1998 through 31 December 2000, federal inspectors collected 98,204 samples and 1,502 completed sample sets for Salmonella analysis from large, small, and very small establishments that produced at least one of seven raw meat and poultry products: broilers, market hogs, cows and bulls, steers and heifers, ground beef, ground chicken, and ground turkey. Salmonella prevalence in most of the product categories was lower after the implementation of PR/HACCP than in pre-PR/HACCP baseline studies and surveys conducted by the FSIS. The results of 3 years of testing at establishments of all sizes combined show that >80% of the sample sets met the following Salmonella prevalence performance standards: 20.0% for broilers, 8.7% for market hogs, 2.7% for cows and bulls, 1.0% for steers and heifers, 7.5% for ground beef, 44.6% for ground chicken, and 49.9% for ground turkey. The decreased Salmonella prevalences may partly reflect industry improvements, such as improved process control, incorporation of antimicrobial interventions, and increased microbial-process control monitoring, in conjunction with PR/HACCP implementation. PMID- 12092727 TI - Microbial profile and antibiotic susceptibility of Campylobacter spp. and Salmonella spp. in broilers processed in air-chilled and immersion-chilled environments. AB - Carcass chilling is considered a critical step for inhibiting bacterial growth during poultry processing. The objective of this study was to compare microbiological loads and the incidence of Salmonella spp. and Campylobacter spp. on broiler carcasses subjected to immersion chilling and air chilling. Additionally, the antibiotic resistance patterns of pathogen isolates were determined. The results of this study indicated that the incidence of Salmonella spp. and Campylobacter spp. tends to be significantly lower in air-chilled broilers, suggesting that cross-contamination may be more prevalent for immersion chilled broilers. No significant differences were detected between chilling treatments for total aerobic populations or for generic E. coli or coliform counts. Psychrotrophic populations were significantly larger (P < 0.05) in immersion-chilled broilers than in their air-chilled counterparts. Campylobacter isolates from immersion-chilled broilers had a higher incidence of resistance to nalidixic acid (NAL) and related fluoroquinolones than isolates from air-chilled broilers did. Additionally, Campylobacter isolates from air-chilled broilers had a higher frequency of resistance to tetracycline than isolates from immersion chilled broilers did. With regard to Salmonella, isolates from immersion-chilled broilers had a higher incidence of resistance to NAL than isolates from air chilled samples did. No Salmonella isolates from immersion- or air-chilled broilers were resistant to the fluoroquinolones tested. The chilling method used during processing may influence the microbial profile of postchilled broilers. PMID- 12092728 TI - Diversity of Campylobacter isolates from retail poultry carcasses and from humans as demonstrated by pulsed-field gel electrophoresis. AB - Campylobacter spp. are a major contaminant of poultry. Eating undercooked chicken and handling raw poultry have been identified as risk factors for campylobacteriosis in humans. Previous studies have found Campylobacter spp. on 90% of poultry carcasses. In the present study, pulsed-field gel electrophoresis (PFGE) was used to assess the genetic diversity of strains on retail poultry carcasses. PFGE patterns of isolates from campylobacteriosis cases were compared to those from the poultry isolates. Over a 1-year study period (March 2000 through February 2001), whole fresh young chickens (n = 72) were obtained from three retail outlets in an urban community in the south-central United States. Campylobacter spp. were isolated from 82% of these carcasses. Strains (n = 70) were defined on the basis of their PFGE pattern. Sixty-seven percent of the carcasses from which Campylobacter spp. were isolated were contaminated with more than one PFGE-distinguishable strain. During the 1-year study period, most of the PFGE patterns (59%) were limited to isolates obtained from a single carcass. Forty-one percent of the PFGE-distinguishable strains were recovered from more than one carcass. Ninety-seven percent of the carcasses contaminated with the same strain were purchased at the same time from the same store. To examine the degree of genetic stability, four strains were followed in vitro over an estimated 1,000 doublings. The PFGE pattern of one of these isolates underwent minor changes during in vitro growth. The data indicate extensive variability in the PFGE patterns of Campylobacter spp. isolated from humans and from poultry carcasses. In spite of difficulties caused by such diversity and the fact that some carcasses are contaminated with more than one strain, the pattern variation provides a useful method for linking a particular strain to its source. PMID- 12092729 TI - Postpackage pasteurization of ready-to-eat deli meats by submersion heating for reduction of Listeria monocytogenes. AB - A mixed cocktail of four strains of Listeria monocytogenes was resuspended in product purge and added to a variety of ready-to-eat (RTE) meat products, including turkey, ham, and roast beef. All products were vacuum sealed in shrink wrap packaging bags, massaged to ensure inoculum distribution, and processed by submersion heating in a precision-controlled steam-injected water bath. Products were run in pairs at various time-temperature combinations in either duplicate or triplicate replications. On various L. monocytogenes-inoculated RTE deli meats, we were able to achieve 2- to 4-log cycle reductions when processed at 195 degrees F (90.6 degrees C), 200 degrees F (93.3 degrees C), or 205 degrees F (96.1 degrees C) when heated from 2 to 10 min. High-level inoculation with L. monocytogenes (approximately 10(7) CFU/ml) ensured that cells infiltrated the least processed surface areas, such as surface cuts, folds, grooves, and skin. D- and z-value determinations were made for the Listeria cocktail resuspended in product purge of each of the three meat categories. However, reduction of L. monocytogenes in product challenge studies showed much less reduction than was observed during the decimal reduction assays and was attributed to a combination of surface phenomena, including surface imperfections, that may shield bacteria from the heat and the migration of chilled purge to the product surface. The current data indicate that minimal heating regimens of 2 min at 195 to 205 degrees F can readily provide 2-log reductions in most RTE deli meats we processed and suggest that this process may be an effective microbial intervention against L. monocytogenes on RTE deli-style meats. PMID- 12092730 TI - Growth and survival of Vibrio parahaemolyticus in postharvest American oysters. AB - Oysters at the retail stage of distribution generally contain greater densities of Vibrio parahaemolyticus than do oysters at harvest. The objective of this study was to determine the effects of postharvest storage at 26 and 3 degrees C on the growth and survival of naturally occurring V. parahaemolyticus in shellstock American oysters (Crassostrea virginica). Oysters were collected monthly from May 1998 through April 1999 from Mobile Bay, Alabama, and their V. parahaemolyticus densities were determined after 0, 5, 10, and 24 h of postharvest storage at 26 degrees C. After 24 h of storage at 26 degrees C, oysters were transferred to a refrigerator at 3 degrees C and analyzed 14 to 17 days later. V. parahaemolyticus numbers were determined by a direct plating method involving an alkaline-phosphatase-labeled DNA probe that targets the species-specific thermolabile hemolysin gene (tlh-AP) to identify suspect isolates. From April to December, when water temperatures at harvest were >20 degrees C, the geometric mean harvest density of V. parahaemolyticus was 130 CFU/g. When water temperatures were <20 degrees C, the geometric mean harvest density was 15 CFU/g. After harvest, V. parahaemolyticus multiplied rapidly in live oysters held at 26 degrees C, showing a 50-fold increase (1.7 log CFU/g) at 10 h and a 790-fold increase (2.9 log CFU/g) at 24 h (April through December). Average V. parahaemolyticus numbers showed a sixfold decrease (0.8 log CFU/g) after approximately 14 days of refrigeration. These results indicate that V. parahaemolyticus can grow rapidly in unrefrigerated oysters. PMID- 12092731 TI - Resistance of cold- and starvation-stressed Vibrio vulnificus to heat and freeze thaw exposure. AB - The effects of cold storage and starvation on the subsequent heat resistance and freeze-thaw resistance of Vibrio vulnificus were studied. Three strains of V. vulnificus were evaluated. Cold stress had no effect on freeze-thaw resistance (P > 0.05). Starvation enhanced freeze-thaw resistance for one strain compared to controls (P < 0.05). V. vulnificus was not heat resistant; control populations were inactivated within 12 min at 47 degrees C. Starvation increased heat tolerance for one strain, but differences were small from a processing perspective (P < 0.05). Cold stress had no effect on heat resistance (P > 0.05). Cold adaptation (holding 4 h at 15 degrees C) enhanced cold temperature (5 degrees C) tolerance. This information will be helpful in the development of methods to minimize V. vulnificus risk. PMID- 12092732 TI - Combined effect of gamma irradiation, ascorbic acid, and edible coating on the improvement of microbial and biochemical characteristics of ground beef. AB - This study was conducted to evaluate the combined effect of gamma irradiation and the incorporation of natural antimicrobial compounds in cross-linked films on the microbiological and biochemical characteristics of ground beef. Medium-fat (23% fat) ground beef patties were divided into three separate treatment groups: (i) control samples without additives, (ii) ground beef samples containing 0.5% (wt/wt) ascorbic acid, and (iii) ground beef samples containing 0.5% ascorbic acid and coated with a protein-based cross-linked film containing immobilized spice powders. Meat samples were irradiated at doses of 0, 1, 2, and 3 kGy and stored at 4 +/- 2 degrees C. Microbial growth (based on total aerobic plate counts [APCs] and total coliforms) was evaluated, as were the content of thiobarbituric acid-reactive substances (TBARS) and that of free sulfydryl groups. At the end of the storage period, Enterobacteriaceae, presumptive Staphylococcus aureus, presumptive Pseudomonas spp., Brochothrix thermosphacta, and lactic acid bacteria were enumerated. Regardless of the treatment group, irradiation significantly (P < or = 0.05) reduced the APCs. Irradiation doses of 1, 2, and 3 kGy produced immediate APC reductions of 2, 3, and 4 log units, respectively. An APC level of 6 log CFU/g was reached after 4, 7, and 10 days for samples irradiated at 1, 2, and 3 kGy, respectively. Lactic acid bacteria and B. thermosphacta were more resistant to irradiation than were Enterobacteriaceae and Pseudomonas. The TBARS and free sulfydryl contents were stabilized during postirradiation storage for samples containing ascorbic acid and coated with the protein-based cross-linked film containing immobilized spice powders. PMID- 12092733 TI - Production of cyclopiazonic acid by Penicillium commune isolated from dry-cured ham on a meat extract-based substrate. AB - Penicillium commune, a mold frequently found on dry-cured meat products, is able to synthesize the mycotoxin cyclopiazonic acid (CPA). To evaluate the hazard due to CPA on such foods, the ability of P. commune to grow and produce CPA at water activities (a(w)) in the range of 0.99 to 0.90 with a meat extract-based medium from 12 to 30 degrees C was determined. CPA was quantified by high-pressure liquid chromatography and mass spectrometry. P. commune was able to grow at every a(w) and temperature tested. The optimal environmental conditions for growth were 20 to 25 degrees C, at 0.97 to 0.96 a(w), but the highest amount of CPA was produced at 30 degrees C, 0.96 a(w). No direct correlation between growth rate and CPA production was assessed. Temperature seems to be the most important factor influencing CPA production. However, there was an interaction between temperature and a(w) that significantly (P < 0.001) affected growth and CPA production. An a(w) of 0.90 had a marked effect, depressing growth and CPA production. Meat extract-based medium proved to be an appropriate substrate for CPA biosynthesis by P. commune under a wide range of conditions. PMID- 12092734 TI - Enumeration and identification of yeasts associated with commercial poultry processing and spoilage of refrigerated broiler carcasses. AB - Yeasts associated with broiler carcasses taken from various stages of commercial poultry processing operations and broiler carcasses stored at refrigerated temperatures were enumerated and identified. Whole carcass rinses were performed to recover yeasts from carcasses taken from a processing facility and processed carcasses stored at 4 degrees C for up to 14 days. Yeasts in the carcass rinsates were enumerated on acidified potato dextrose agar and identified with the MIDI Sherlock Microbial Identification System. Dendrograms of fatty acid profiles of yeast were prepared to determine the degree of relatedness of the yeast isolates. Findings indicated that as the carcasses are moved through the processing line, significant decreases in the number of yeasts associated with broiler carcasses usually occur, and the composition of the yeast flora of the carcasses is altered. Significant (P < 0.05) increases in the yeast population of the carcasses generally occur during storage at 4 degrees C, however. Furthermore, it was determined that the same strain of yeast may be recovered from different carcasses at different points in the processing line and that the same strain of yeast may be isolated from carcasses processed on different days in the same processing facility. PMID- 12092735 TI - Concentration and detection of caliciviruses from food contact surfaces. AB - Outbreaks of human Norwalk virus (NV) and Norwalk-like viruses often originate in food service establishments. No reliable method is available for the detection of these human caliciviruses on food contact surfaces. We describe a simple method for the detection of NV from stainless steel work surfaces using cultivable feline calicivirus (FCV) as a model. Stainless steel surfaces were artificially contaminated with known amounts of FCV, followed by its elution in a buffer solution. Three methods of virus elution were compared. In the first method, moistened cotton swabs or pieces of nylon filter (1MDS) were used to elute the contaminating virus. The second method consisted of flooding the contaminated surface with eluting buffer, allowing it to stay in contact for 15 min, followed by aspiration of the buffer (aspiration method) after a contact period of 15 min. The third method, the scraping-aspiration method, was similar to the aspiration method, except that the surfaces were scraped with a cell scraper before buffer aspiration. Maximum virus recovery (32 to 71%) was obtained with the scraping aspiration method using 0.05 M glycine buffer at pH 6.5. Two methods (organic flocculation and filter adsorption elution) were compared to reduce the volume of the eluate recovered from larger surfaces. The organic flocculation method gave an average overall recovery of 55% compared to the filter-adsorption-elution method, which yielded an average recovery of only 8%. The newly developed method was validated for the detection of NV by artificial contamination of 929-cm2 stainless steel sheets with NV-positive stool samples and for the detection of the recovered virus by reverse transcription-polymerase chain reaction. PMID- 12092736 TI - Design and operation of the National Survey of Early Childhood Health, 2000. AB - OBJECTIVES: This report presents the development, plan, and operation of the National Survey of Early Childhood Health, a module of the State and Local Area Integrated Telephone Survey, conducted by the National Center for Health Statistics, Centers for Disease Control and Prevention. This survey was designed to assess parents' perceptions of their children's pediatric care. In addition, data were collected that can be used to examine relationships between the promotion of health in the pediatric office and promotion of health in the home. Funding for the survey was provided by The Gerber Foundation, the American Academy of Pediatrics, and the Maternal and Child Health Bureau, Health Resources and Services Administration. The UCLA Center for Healthier Children, Families, and Communities contributed to the design of the study and the questionnaire. METHODS: A national random-digit-dialed (RDD) sample of households with children 4-35 months of age was selected. The study included an oversample of households having an eligible black non-Hispanic or Hispanic child. In households with more than one eligible child, one was randomly selected to be the subject of the interview. The respondent was the parent or guardian who was most responsible for the child's health care. A computer-assisted telephone interviewing (CATI) system was used to collect the data. RESULTS: A total of 2,068 interviews were completed during the first half of 2000. The response rate was 65.6%. A data file has been released that contains demographic information on the focal child and respondent, substantive health and health-related data, and sampling weights. Estimates based on the sampling weights generalize to the entire U.S. population of children 4-35 months of age. PMID- 12092738 TI - Advanced science education in the regulatory arena: the Center for Drug Evaluation and Research Experience at the Food and Drug Administration. AB - A challenge faced by the Center for Drug Evaluation and Research (CDER) in effectively carrying out its mission requires it to integrate the disciplines of science, medicine, law, and public policy. One way to do that is by ensuring a highly trained multidisciplinary staff. The CDER has been able to meet this requirement by identifying the core competencies needed to accomplish its mission. The use of a competency-based training model in the planning, development, and delivery of its advanced scientific education program allows CDER staff to maintain current knowledge as well as prepare for future scientific education needs. The CDER educational model could be readily adapted to meet the educational needs of other organizations. PMID- 12092737 TI - Trends of medication errors in hospitalized children. AB - Medication errors are a major cause of morbidity and mortality among hospitalized children. Due to the small volumes of stock solution involved, even a large error may look as an unsuspiciously small dose. Strategies were implemented to reduce medication errors in a large tertiary pediatric hospital in Toronto. Starting in 1993, several initiatives were taken, including a new hospital computer system for medication ordering, a review process to remove hazardous drugs from wards where they are not needed immediately, and in the training of pediatric residents. The rates of reported medication errors were compared before and after these initiatives were taken. Compared to baseline, there was a steady and a statistically significant decrease in medication errors through the decade. Total errors (actual and potential) decreased for nurses and physicians by half and for pharmacists by 75%. Actual incidents decreased by half. Moderate and severe errors decreased by more than 70%. It was concluded that a combination of several initiatives to decrease system and human errors has resulted in more than a 50% reduction of medication errors reaching the pediatric patient. PMID- 12092739 TI - Educational issues in clinical pharmacology: who are our audiences and what are their specialized needs? One specialized need: "understanding the role of veterinary medicine in public health". AB - When considering educational issues and the need to update the curriculum for clinical pharmacologists for the new millennium, a number of questions must be raised. Who are our audiences? What are the specialized needs? This educational article identifies the audience, which includes those with diverse degrees such as MDs, PhDs, PharmDs, RNs, DVMs, and other non-MD prescribers working in academia, industry, clinical research organizations, and government in multifaceted disciplines requiring a knowledge base of physiology, pharmacology, biochemistry, anatomy, microbiology, pathology, medicine, and the drug development process of preclinical and clinical studies complete with protocols, pharmacokinetics, and statistics. One specialized current educational issue for clinical pharmacologists to understand is the impact of animal therapeutic and subtherapeutic use of antimicrobials on antibiotic use in human medicine. PMID- 12092740 TI - Systemic availability and pharmacokinetics of thymol in humans. AB - Essential oil compounds such as found in thyme extract are established for the therapy of chronic and acute bronchitis. Various pharmacodynamic activities for thyme extract and the essential thyme oil, respectively, have been demonstrated in vitro, but availability of these compounds in the respective target organs has not been proven. Thus, investigation of absorption, distribution, metabolism, and excretion are necessary to provide the link between in vitro effects and in vivo studies. To determine the systemic availability and the pharmacokinetics of thymol after oral application to humans, a clinical trial was carried out in 12 healthy volunteers. Each subject received a single dose of a Bronchipret TP tablet, which is equivalent to 1.08 mg thymol. No thymol could be detected in plasma or urine. However, the metabolites thymol sulfate and thymol glucuronide were found in urine and identified by LC-MS/MS. Plasma and urine samples were analyzed after enzymatic hydrolysis of the metabolites by headspace solid-phase microextraction prior to GC analysis and flame ionization detection. Thymol sulfate, but not thymol glucuronide, was detectable in plasma. Peak plasma concentrations were 93.1+/-24.5 ng ml(-1) and were reached after 2.0+/-0.8 hours. The mean terminal elimination half-life was 10.2 hours. Thymol sulfate was detectable up to 41 hours after administration. Urinary excretion could be followed over 24 hours. The amount of both thymol sulfate and glucuronide excreted in 24-hour urine was 16.2%+/-4.5% of the dose. PMID- 12092741 TI - Safety, tolerability, pharmacokinetics, and time course of pharmacologic response of the active metabolite of roxifiban, XV459, a glycoprotein IIb/IIIa antagonist, following oral administration in healthy volunteers. AB - Roxifiban is an esterprodrug that is hydrolyzed, after oral administration, to the active glycoprotein (GP) IIb/IIIa antagonist, XV459. The objectives of the study were to investigate the safety, tolerability, pharmacokinetics, and the time course of the pharmacologic response of XV459 in escalating doses of roxifiban and to assess the effect of age, loading dose of roxifiban, and aspirin pretreatment on XV459 pharmacokinetics, pharmacologic response, and safety profile in a five-part double-blind, placebo-controlled study. Healthy male volunteers (ages 18-46 years) received 7 (0.75-1.5 mg; n = 20) and 10 (0.75-1.0 mg; n = 8) multiple, oral, qd doses of roxifiban or placebo (n = 5). Healthy older male and female volunteers (ages 47-75 years) received roxifiban qd doses (0.5-0.75 mg; n = 8) or placebo (n = 3) for 7 days. Healthy male subjects (ages 18-46 years; n = 16) received a 1.5 or 1.0 mg loading dose either with or without pretreatment of 325 mg aspirin once daily for 3 days followed by single daily doses of 1.0 mg roxifiban for 6 days. Measurable plasma concentrations of XV459 appeared rapidly and were sustained throughout the dosing interval of 24 hours. The pharmacokinetics of XV459 were nonlinear. Systemic exposure of XV459 plateaued at the 1-mg dose level; plasma concentrations approached steady state in 4 to 6 days for doses greater than 1.0 mg. The time course of pharmacologic response as measured by the inhibition of platelet aggregation in response to an ex vivo 10 microM adenosine 5'-diphosphate (ADP) agonist correlated closely to the plasma concentration of XV459. Potent inhibition of ADP-induced platelet aggregation (IPA) persisted over the entire dosing interval. A clear dose response was achieved with roxifiban doses of 0.5 and 1.0 mg. For doses greater than 1.0 mg, a dose-proportional increase in IPA was not observed. Both the pharmacokinetics and pharmacologic response of XV459 exhibited low intraindividual variability (coefficient of variation [CV] < 15%) and higher interindividual variability (CV < 30%). Pretreatment with aspirin and/or a loading dose of 1.5 mg roxifiban had no significant effect on the pharmacokinetics and pharmacologic response of XV459. A dose-related increase in template bleeding time was observed at 1.25- and 1.5-mg doses of roxifiban, as compared to placebo. However, these bleeding time increases in the 1.25- and 1.5 mg dose groups were not significantly different from those at the lower dose groups. Overall, once-daily oral administration of roxifiban was fairly well tolerated and provided sustained systemic drug exposure and pharmacologic response over the entire administration interval. PMID- 12092742 TI - Comparative effects on dynamic renal potassium excretion of ACE inhibition versus angiotensin receptor blockade in hypertensive patients with type II diabetes mellitus. AB - Both ACE inhibitors (ACEI) and angiotensin receptor blockers (ARB) are renoprotective beyond their effects on blood pressure (BP), but their widespread use is limited by their tendency to provoke hyperkalemia. The comparative effects of ACEI and ARB on potassium handling have not been investigated. The objective of this study was to determine whether there are differences in dynamic renal potassium handling between ACEI and ARB in response to an oral potassium challenge. This was a randomized crossover study of candesartan versus lisinopril titrated to control BP followed by an inpatient study of renal potassium handling in 24 hypertensive patients with type II diabetes mellitus (DMII) and preserved renal function. Following an oral potassium challenge (0.75 mmol/kg), differences in hourly serum K (mmol/L), rate of urinary potassium excretion (UkV, micromol/min), and fractional excretion of potassium (FEK) were assessed by repeated-measures ANOVA. Hourly UkV(p = .45) and FEK (p = .19) were similar for candesartan and lisinopril, although FEK at 2 hours for candesartan tended to exceed that for lisinopril (.34 [.04] vs. .26 [.03]) and approached significance (p = .096). UkVfor candesartan at hour 2 was 177 (26) and 121 (21) for lisinopril and also approached significance (p = .10). Serial serum potassium did not differ (p = .70). No statistical differences were discovered in renal potassium handling between candesartan and lisinopril in patients with DMII and preserved renal function. Whether there are differences between the drug classes in renal impairment remains to be determined. PMID- 12092744 TI - Pharmacokinetics of histamine dihydrochloride in healthy volunteers and cancer patients: implications for combined immunotherapy with interleukin-2. AB - Recent clinical trials in melanoma and leukemia have demonstrated potential for increased survival time and improved remission when histamine dihydrochloride is added to cytokine monotherapy. In the present study, the pharmacokinetics of subcutaneous histamine (1 mg) in 21 healthy subjects and 12 melanoma patients was determined via model-dependent methods. Drug-drug interactions with subcutaneous interleukin-2 (1.1 mg) were evaluated in a combined cohort of patients with melanoma (n = 8) or renal cell carcinoma (n = 4). Histamine dihydrochloride administered over 10 minutes in healthy subjects peaked at 18 minutes (Cmax 38 nmol/L), attained a distribution volume of 59 L, and was eliminated at 6%/min. The results were similar in a 20-minute infusion in melanoma patients. No gender effects were observed (p > 0.05). Interleukin-2 injected either 10 minutes prior to or 10 minutes following histamine dihydrochloride had no effect on histamine kinetics. Histamine dihydrochloride administered 10 minutes prior to injection of interleukin-2 also had no effect on interleukin-2 kinetics. Maximal concentration of interleukin-2 (2,442 pg/ml) occurred at 2.5 hours with an elimination half life of 1.7 hours, area under the curve (AUC) of 15,746 pg x h/ml, and volume of distribution and plasma clearance of 194 L and 74 L/h, respectively. However, interleukin-2 Cmax (1,758 pg/ml) and AUC (12,448 pg x h/ml) were reduced when histamine dihydrochloride was infused 10 minutes following interleukin-2, likely due to the pharmacodynamic effects of histamine, including increased heart rate and reduced blood pressure. It is concluded that histamine dihydrochloride and interleukin-2 can be safely coadministered with minimal interaction. PMID- 12092743 TI - Toxicity of high-dose carboplatin: ultrafiltered and not total plasma pharmacokinetics is of clinical relevance. AB - Combination chemotherapy containing high-dose carboplatin is a therapeutic option for patients with poor-risk germ cell tumors. Since such treatment is relatively toxic, pharmacokinetic characteristics of total and ultrafiltered platinum, representing different species of platinum complexes formed, were investigated in 29 patients in relation to toxicity. The goodness-of-fit parameters revealed that a two-compartment model with weighting (1/C2i) resulted in the best fit. The terminal half-life of total platinum > 5 days was fourfold longer than that of ultrafiltered platinum, indicating long-term retention. At stem cell rescue, an ultrafiltered concentration of < or = 0.223 microg/ml did not impair hematological recovery. Ultrafiltered but not total platinum concentrations or AUC values were significantly correlated to an increase in creatinine concentration and different types of toxicity associated with high-dose treatment. Since the fraction unbound is time dependent and highly variable among patients, measuring ultrafiltered concentrations is highly recommended as they represent the clinically relevant platinum complexes in high-dose treatment. PMID- 12092746 TI - Bioequivalence of two formulations of didanosine, encapsulated enteric-coated beads and buffered tablet, in healthy volunteers and HIV-infected subjects. AB - Didanosine is an acid labile drug and hence has been given with buffering agents. To avoid the need for concurrent administration with antacids, an encapsulated enteric-coated bead formulation of didanosine was developed. The objective of this study was to assess the bioequivalence of the encapsulated enteric-coated beads compared to the buffered tablet. Two separate open-label, randomized, two way crossover studies were conducted, one in healthy subjects and the other in HIV-infected subjects (with CD4 cell counts > 200 cells/mm3). All subjects received a 400-mg dose of the buffered tablet (reference formulation) and the encapsulated enteric-coated beads (test formulation). Blood samples were collected over 12 hours, and plasma levels of didanosine were determined using a validated assay. The 90% confidence interval (CI) of the ratio of the geometric means of log-transformed Cmax and AUCinfinity values were used to assess bioequivalence between the two formulations using the equivalence interval of 0.80 and 1.25. In healthy volunteers (n = 46), the point estimate and 90% CI of the ratios of Cmax and AUCinfinity values were 0.58 (0.52, 0.64) and 1.02 (0.95, 1.01), respectively. In HIV-infected subjects (n = 30), the point estimate and 90% CI of the ratios of Cmax and AUCinfinity values were 0.64 (0.56, 0.72) and 0.95 (0.86, 1.06), respectively. Median t(max) value increased significantly from 0.67 hours for the buffered tablet in both studies to 2.33 hours (in healthy subjects) or 2.0 hours (in HIV-infected subjects) for the enteric-coated beads. The mean half-life of didanosine was similar between treatments and ranged between 1.60 and 1.70 hours across healthy and HIV-infected subjects. It was concluded that the encapsulated enteric-coated bead formulation of didanosine is equivalent to the buffered tablet in the extent of exposure but differs in the rate of absorption. The pharmacokinetic profile of the enteric formulation appears to be similar in healthy and HIV-infected subjects. PMID- 12092745 TI - Pharmacokinetic and pharmacodynamic properties of a new thromboxane receptor antagonist (Z-335) after single and multiple oral administrations to healthy volunteers. AB - The pharmacokinetics and pharmacodynamics of a new oral thromboxane (TX) A2 receptor antagonist, Z-335, were studied in healthy male volunteers following single doses (0.5-40 mg, PO) in a dose-escalating manner and multiple doses (40 mg, PO, once daily for 7 consecutive days) with a single-blind, placebo controlled design. Serial blood and urine samples were analyzed for Z-335 and its metabolites to obtain key pharmacokinetic parameters. In the single-dose (10, 20, and 40 mg) study, the maximum plasma concentration (Cmax) and area under the plasma concentration versus time curve (AUC) increased in proportion to the dose when administered afterfasting, while the mean elimination half-life (t1/2beta) was essentially unchanged (7.79-7.93 h). Recovery of the unchanged and taurine conjugated drugs in the urine within 24 hours was 6.5% to 8.4% and 11.9% to 14.2%, respectively. These parameters essentially remained unchanged when the effect of meal intake was evaluated at the dose of 20 mg with a crossover design. Ex vivo platelet aggregation in the plasma by a TXA2 analogue, U46619, was completely inhibited within 2 hours after all doses, and complete inhibition was maintained for 12 to 14 hours, depending on the dose. The aggregation induced by collagen was also inhibited to a lesser extent, whereas that by adenosine diphosphate was hardly influenced. In the multiple-dose study, Cmax and AUC0-24 were increased by 34% after the last dose compared with the first dose. Z-335 afforded extensive inhibition of platelet aggregation by U46619 throughout the administration period, which returned, however, almost to the control level 48 hours after the last dose. The agent was well tolerated without any abnormalities in subjective and objective symptoms, blood biochemistry, hematology, and urinalysis definitely attributable to the agent, except for the changes expected from its TXA2 receptor-antagonizing actions. Z-335 was concluded to be safe and to provide long-lasting blockade of TXA2 receptors on the basis of a once-daily regimen, promoting further clinical evaluation. PMID- 12092747 TI - Evaluation of renal function in transplant patients on tacrolimus therapy. AB - Glomerular filtration rate (GFR), as measured by 24-hour creatinine clearance and clearance of iothalamate, and effective renal plasma flow (ERPF), as measured by the clearance of para-aminohippuric acid (PAH), were evaluated at 2 weeks, 1 month, and 3 months after transplantation in 8 renal transplant patients and at 1 month and 1 year after transplantation in 9 liver transplant patients receiving tacrolimus (Prograf) therapy. In renal transplant patients, there was a significant increase in GFR after transplantation. There was no change in GFR at 1 and 3 months as compared to 2 weeks after transplantation, while ERPF (ml/min/1.73 m2) was lower (p < 0.05) at 3 months (212+/-42) compared to 1 month (306+/-118) after transplantation. In liver transplant patients, GFR and ERPF were below normal despite normal serum creatinine concentrations, but there was no difference in GFR or ERPF at 1 month and 1 year after transplantation. Although below normal, renal function was well preserved in transplant patients while receiving chronic tacrolimus therapy over the study period. Dosage alterations ofrenally eliminated drugs may be required for drugs with a narrow therapeutic index. PMID- 12092748 TI - Impairment of ciprofloxacin absorption by calcium polycarbophil. AB - The effect of calcium polycarbophil on the absorption of ciprofloxacin, a broad spectrum antibacterial agent, was evaluated in an in vitro and in vivo study. In the in vitro study, the release of ciprofloxacin from the cellulose membrane in the presence or absence of metal cations was measured using the dissolution test procedure. In the in vivo study, male ST Wistar rats and male volunteers were employed. First, 20 mg/kg of ciprofloxacin alone (Rat Study 1) or 20 mg/kg of ciprofloxacin in combination with 64 mg/kg of calcium chloride (Rat Study 2) was administered orally to 3 rats. Second, a volunteer study was employed and a randomized crossover design with twophases was used. In onephase, volunteers received 400 mg of ciprofloxacin alone (Study 1); in the other phase, they received 400 mg of ciprofloxacin and 1200 mg of fine calcium polycarbophil granules concomitantly (Study 2). The plasma and serum concentrations of ciprofloxacin were measured by high-performance liquid chromatography. The release of ciprofloxacin from the cellulose membrane in the presence of aluminum, calcium, or iron ions was slower than that in the absence of these metal ions. The AUC0-4 and Cmax in Rat Study 2 were lower than those respective values in Rat Study 1. AUC0-4 was approximately 60% lower in Rat Study 2 than Rat Study 1. In the volunteer study, the AUC0-12 and Cmax in Study 2 were lower than those respective values in Study 1. In particular, AUC0-12 was approximately 50% lowerin Study 2 than in Study 1. These findings suggest that when ciprofloxacin and calcium polycarbophil were coadministered concomitantly, a decrease of ciprofloxacin absorption was observed, and this action was caused by the formation of chelate complexes. Therefore, it seems clear that we should avoid the concomitant administration of ciprofloxacin and calcium polycarbophil. PMID- 12092749 TI - Utility of chromosomal position of heterochromatin as a biomarker of radiation induced genetic damage: a study of Chornobyl voles (Microtus sp.). AB - Biomarkers that effectively document effects of chronic multi-generational exposure to contaminated environments on chromosomes would be valuable in risk assessment, remediation, and environmental decisions. Native, free-ranging populations of voles inhabiting the highly radioactive regions surrounding Reactor 4 of the Chornobyl Nuclear Power Station provide a model system to evaluate biological and chromosomal effects of chronic multi-generational exposure to radioactivity and other reactor meltdown-related pollutants. Here, we explore the utility of heterochromatic elements as potentially informative biomarkers for genetic damage in voles from the radioactive environments surrounding Chornobyl. We analyzed chromosomal positions of heterochromatin from Microtus arvalis and M. rossiaemeridionalis using fluorescent in situ hybridization. Although intrapopulational variation existed in chromosomal position and abundance of heterochromatin, none of that variation could be assigned to environmental exposure. PMID- 12092750 TI - Accumulation of lipophilic microcontaminants and biochemical responses in eels from the Camargue Biosphere Reserve. AB - We assessed the impacts of persistent organic micropollutants on aquatic trophic webs in brackish and freshwater communities in the Camargue National Nature Reserve (NNR). We found that organochlorine compounds affect fish communities, particularly those of the common eel (Anguilla anguilla). The aims of this study were (1) to determine the amount of lipophilic xenobiotics such as polycyclic aromatic hydrocarbons (PAHs), which are probably of atmospheric origin, and two organochlorines (OC), lindane and dieldrin, which are from irrigation waters, in liver and muscle (2) to define in situ biomarkers and (3) to identify the mode by which fish from 'unpolluted' areas become contaminated. All of the species were contaminated with low, but fluctuating quantities of PAHs and OCs, regardless of the sampling season. Lindane and dieldrin were always detected and naphtalene was the most abundant hydrocarbon. The OC and PAH content was rarely correlated with the lipid content in storage tissues and their concentrations in the lipidic fractions (neutral and polar) varied greatly. We found a number of correlations between persistent organic pollutant (POP) tissue concentrations and the activities of enzymatic membrane markers. For example, there is a relationship between the concentrations of the most volatile PAHs and the activity of muscle acetylcholinesterase (AChE) and between the concentration of benzo-PAH and the activities of ATPases in the gills and/or muscle. PMID- 12092752 TI - Effects of a mixture of two insecticides in freshwater microcosms: II. Responses of plankton and ecological risk assessment. AB - This paper reports on the chronic effects of a mixture of the insecticides chlorpyrifos and lindane in freshwater microcosms. Chronic treatment levels corresponding to concentrations of 0, 0.005, 0.01, 0.05, 0.1 and 0.5 times the LC50 of the most sensitive standard test organism were evaluated. The zooplankton community structure was altered from the 0.05 * LC50 treatment level upwards. Cladocerans were the most susceptible group, followed by Copepoda and Ostracoda. Rotifera increased in abundance at the higher treatment levels. Increased abundance of some phytoplankton taxa and increased chlorophyll-a levels were found at the two highest treatment levels, most probably a consequence of decreased grazing pressure. Threshold levels for the mixture, both at population and community/ecosystem level, corresponded well with those reported in the literature for the individual compounds. The overall risk assessment indicates no antagonistic or synergistic effects of the mixture at ecosystem level. It was found that the safety factors set by the Uniform Principles for individual compounds also ensure protection against chronic exposure to a mixture of insecticides at community level, though not always at species level. PMID- 12092751 TI - Effects of a mixture of two insecticides in freshwater microcosms: I. Fate of chlorpyrifos and lindane and responses of macroinvertebrates. AB - Effects of chronic application of a mixture of the insecticides chlorpyrifos and lindane were studied in indoor freshwater microcosms. The exposure concentrations (based on 0, 0.005, 0.01, 0.05, 0.1 and 0.5 times the LC50 of the most sensitive standard test organism for each compound) were kept at a constant level for four weeks. The calculated mean concentrations for chlorpyrifos were found to be almost at their corresponding nominal level during the treatment period. The mean calculated lindane concentrations, however, were found to be 15-40% higher than intended. In the post treatment period both insecticides dissipated fast (t 1/2: chlorpyrifos 9 days, lindane 22 days) from the water phase. The concentrations of the mixture at the highest treatment level corresponded to 0.53 toxic units (TU) for Daphnia magna and 0.61 TU for the most sensitive fish. The decomposition of Populus leaves in litter bags was significantly lower at the three highest insecticide concentrations. The macroinvertebrate community was seriously affected at the three highest treatment levels, with Crustacea and the Chironomidae Corynoneura proving to be the most sensitive groups. Gastropoda and Oligochaeta were relatively insensitive and some taxa (e.g. Valvata piscinalis, juvenile Physa fontinalis, Nemertea and Stylaria lacustris) increased in numbers. The observed effects could be explained from the individual toxicity of the insecticides to the invertebrates, and did not indicate synergistic effects. A second paper (Van den Brink et al., 2002) addresses the effects on other endpoints, as well as the overall risk assessment of the insecticide mixture. PMID- 12092753 TI - The ecological and physiological costs of lead shot and immunological challenge to developing western bluebirds. AB - We investigated the interacting effects of a nonpathogenic immunological challenge and exposure to lead shot early in the development of nestling western bluebirds (Sialia mexicana). Nestlings were randomly assigned to each of six treatments of an incomplete block design with two antigen treatments, Newcastle disease virus (NDV) or sheep red blood cells (SRBC), and four lead shot treatments (no shot, one shot--0.05 g, 2 shots--0.1 g, or three shots--0.15 g). Survival functions did not differ between the lead treatment groups, and there were no effects of lead treatments on weight, growth rates, fluctuating asymmetry (FA), or antibody response. NDV- and SRBC-treated birds survived better than control birds which may be due to an adjuvant-activation of the entire immune system. However, FA was greater in individuals in the NDV and SRBC treatment groups, suggesting a tradeoff between growth and immunocompetence. Cell-mediated response to phytohemagglutinin of the high-lead treatment groups was significantly less than other groups. Hematocrit increased with age and weight, and was not affected by lead or antigen. While in this study the shape of the growth curve, FA, cell-mediated immunity, and behavior were affected by the higher dose of lead shot, actual lead concentrations in blood are needed to verify this process. Antigenic stress was documented in nestling developmental stability and there were no measured effect of both the lead shot and immunological challenge in combination. PMID- 12092754 TI - Analyses of the recycling potential of medical plastic wastes. AB - This study analyzed the recycling potential of plastic wastes generated by health care facilities. For this study, we obtained waste streams and recycling data from five typical city hospitals and medical centers and three animal hospitals in Massachusetts. We analyzed the sources, disposal costs and plastic content of medical wastes, and also determined the components, sources, types and amounts of medical plastic wastes. We then evaluated the recycling potential of plastic wastes produced by general city hospital departments, such as cafeterias, operating rooms, laboratories, emergency rooms, ambulance service and facilities, and animal hospitals. Facilities, laboratories, operating rooms, and cafeterias were identified as major sources of plastic wastes generated by hospitals. It was determined that the recycling potential of plastics generated in hospital cafeterias was much greater than that in other departments. This was mainly due to a very slight chance of contamination or infection and simplification of purchasing plastic components. Finally, we discuss methods to increase the recycling of medical plastic wastes. This study suggests that a classification at waste generating sources, depending upon infection chance and/or plastic component, could be a method for the improved recycling of plastic wastes in hospitals. PMID- 12092755 TI - Peanut hull pellets as a single use sorbent for the capture of Cu(II) from wastewater. AB - This paper investigates the ability of peanut hulls and peanut hull pellets to adsorb copper from dilute aqueous metal ion solutions in batch and fixed bed systems. The kinetics of copper uptake onto the media have been investigated in batch systems and the influence of pH and particle size on the rate and extent of copper capture determined. The Langmuir and Freundlich isotherm theories were determined; the Langmuir model was found to best represent the equilibrium isotherm data. In normalized kinetic tests at least 75% of copper removal occurred within the first 20 min; 92% removal was effected within the first 50 min. The rate of uptake was optimum within the pH range 5-7.5, and media capacities remained relatively constant at a pH above 4.0. Bench-scale column studies were performed using peanut hull pellets. The overall capacity of pelletized peanut hulls was higher than for unmodified peanut hulls. Due to their demonstrated ability for Cu(II) uptake and favorable structural characteristics, pelletized peanut hulls could gain use as a low-cost, once-through biomass filter medium for copper-bearing waste streams. PMID- 12092756 TI - Immobilization of radioactive waste by cementation with purified kaolin clay. AB - A study is undertaken to determine the waste immobilization performance of low level wastes in cement-clay mixtures. Liquid low-level wastes are precipitated using chemical methods, followed by solidification in drums. Solidification is done using cementation processes. Long-term leaching rates of the radionuclides are used as indicators of immobilization performance of solidified waste forms. In addition to evaluating the effects of kaolin clay on the leaching properties of the cemented waste forms, the effect of addition of kaolin on the strength of the cemented waste form is also investigated. The long term leaching tests show that inclusion of kaolin in cement reduces the leaching rates of the radionuclides significantly. However, clay additions in excess of 15 wt.% causes a significant decrease in the hydrolytic stability of cemented waste form. It is found that the best waste isolation, without causing a loss in the mechanical strength, is obtained when the kaolin content in cement is 5%. PMID- 12092757 TI - Treatment and recycling of incinerated ash using thermal plasma technology. AB - To treat incinerated ash is an important issue in Taiwan. Incinerated ashes contain a considerable amount of hazardous materials such as dioxins and heavy metals. If these hazardous materials are improperly treated or disposed of, they shall cause detrimental secondary contamination. Thermal plasma vitrification is a robust technology to treat and recycle the ash residues. Under the high temperature plasma environment, incinerated ashes are vitrified into benign slag with large volume reduction and extreme detoxification. Several one-step heat treatment processes are carried out at four temperatures (i.e. 850, 950, 1,050 and 1,150 degrees C) to obtain various "microstructure materials". The major phase to form these materials is a solid solution of gehlenite (Ca2Al2SiO7) and akermanite (Ca2MgSi2O7) belonging to the melilite group. The physical and mechanical properties of the microstructure materials are improved by using one step post-heat treatment process after plasma vitrification. These microstructure materials with good quality have great potential to serve as a viable alternative for construction applications. PMID- 12092758 TI - Electrocoagulation of some reactive dyes: a statistical investigation of some electrochemical variables. AB - In this study, the decolorization of strong colored solutions containing the reactive textile dyes by electrocoagulation was investigated. The experiments were statistically designed and carried out according to a 2(4) full factorial design with two replicate and four center points. This design was extended with eight additional axial points. Then, the non-linear regression was applied on the data by using MINITAB software. The variables chosen for this work are mixing rate, cell voltage, electrolysis time and current density. Results showed that the effective variables on decolorization process are cell voltage, electrolysis time and current density. In addition, to determine the effect of dye and electrode type on decolorization, the additional experiments were carried out. It was determined the dye and electrode type are important on the decolorization process. PMID- 12092759 TI - Development of a purpose built landfill system for the control of methane emissions from municipal solid waste. AB - In the present paper, a new system of purpose built landfill (PBLF) has been proposed for the control of methane emissions from municipal solid waste (MSW), by considering all favourable conditions for improved methane generation in tropical climates. Based on certain theoretical considerations multivariate functional models (MFMs) are developed to estimate methane mitigation and energy generating potential of the proposed system. Comparison was made between the existing waste management system and proposed PBLF system. It has been found that the proposed methodology not only controlled methane emissions to the atmosphere but also could yield considerable energy in terms of landfill gas (LFG). Economic feasibility of the proposed system has been tested by comparing unit cost of waste disposal in conventional as well as PBLF systems. In a case study of MSW management in Mumbai (INDIA), it was found that the unit cost of waste disposal with PBLF system is seven times lesser than that of the conventional waste management system. The proposed system showed promising energy generation potential with production of methane worth of Rs. 244 millions/y ($5.2 million/y). Thus, the new waste management methodology could give an adaptable solution for the conflict between development, environmental degradation and natural resources depletion. PMID- 12092760 TI - Using organoclays to enhance carbon filtration. AB - Organoclays have found increased acceptance as pre-treatment for activated carbon adsorption systems in both groundwater and wastewater cleanup. The reason is that activated carbon tends to become quickly blinded by large organic molecules of low solubility, particularly oils. However, it is also well established that activated carbon is more efficient at low concentrations of organic contaminants than at higher ones, i.e. at less than 1 ppm. With organoclays it is exactly the opposite, they are better at removing organics at higher concentrations, above 3 ppm. Therefore it is cost effective in these applications to use two or more vessels in series, the first one filled with organoclay, the remainder with activated carbon. The economics make sense, even though the organoclay is not regenerated, because of the reduction in down time every time a carbon vessel has to be changed out. Use of organoclays increases the volume treated by carbon in many applications seven to nine fold. In the case of other organic contaminants, as the aqueous solubility increases, the efficiency decreases, except in the case of methylene chloride, which it removes at far higher efficiency then carbon. This article presents the results of a series of tests, including Kd determinations, jar tests, and mini-column tests. These tests determined the adsorption capacity and efficiency of organoclay and activated carbon for the removal of benzene, toluene, xylene and naphtalene from water. These tests were followed by adding the four compounds into one container to see if the combination of organoclay, followed by carbon, would be more efficient then each sorbent alone. The tests also compared the efficiency of organoclay versus carbon for the removal of various oils from water. PMID- 12092761 TI - Processing of Egyptian boiler-ash for extraction of vanadium and nickel. AB - Proposed technique in this investigation is given for vanadium and nickel enrichment in the Egyptian boiler ash. Among the various concepts for recovery of vanadium and nickel from boiler ash, the pyro-metallurgical approach is technically feasible, but is not cost-effective from an operational economy standpoint. Another technically viable process which, however, needs further development and presented in this investigation, is the hydrometallurgical processing that involves acid leaching under oxygen pressure of ground ash, followed by electrolytic separation of nickel from sulphate solution and vanadium is then neutralized and precipitated by adjustment the pH value and calcined to produce V2O5. PMID- 12092762 TI - Brick production with dredged harbour sediments. An industrial-scale experiment. AB - A volume of 600.000 m3 harbour sediments is annually dredged out of the harbour basin of Bremen to maintain a certain water depth. Because of its perpetual availability, homogeneity and mineralogical, petrographic and chemical composition, the sediment is regarded as a suitable raw material for brick production. A pilot experiment was conducted at a full-scale industrial brickworks. During production, the environmental standards concerning waste-water treatment and the quality of exhausted gas were sufficiently fulfilled. Bricks specified as "building bricks" were produced according to German industrial standards. The parameters pH-value and grain size were varied in leaching tests performed on the bricks as both parameters are likely to change in the course of the brick's life cycle. The leaching data showed that As was stabilised and heavy metals were immobilised in a way that the bricks were not (hazardous to soil or groundwater) neither by their use, for example, in masonry, nor afterwards, when they will be deposited as mineral demolition mass. PMID- 12092763 TI - Process identification and model development of contaminant transport in MSWI bottom ash. AB - In this work we investigate to what extent we are able to predict experimental data on column leaching of heavy metals from municipal solid waste incinerator (MSWI) bottom ash, using the current knowledge on processes controlling aqueous heavy metal concentrations in combination with a multicomponent reactive transport computer model. Heavy metal concentrations were modelled with a surface complexation model for metal sorption to (hydr)oxide minerals in the bottom ash matrix. For transport modelling it was necessary to simplify the sorption modelling approach. Therefore, we determined a minimal set of components and species that still provided an adequate description of the pH dependent heavy metal behaviour. The concentration levels of the heavy metals are generally predicted to within one order of magnitude. Discrepancies between the model and the data are caused by uncertainty in modelling parameters and a still insufficient description of the dynamics of macroelement leaching and pH. In general, the simulated leaching curves show much more abrupt changes than the measurements. This observation might be an indication of non-equilibrium. Processes that have to be taken into account for further model development are the influence of non-equilibrium effects and the facilitated transport of heavy metals by dissolved organic matter. PMID- 12092765 TI - Immobilisation of beryllium in solid waste (red-mud) by fixation and vitrification. AB - The objective of this study was to obtain information on the immobilization of beryllium (Be) in solid waste generated in the extraction process of beryllium from its ore, Beryl. This solid waste, termed red-mud, contains oxides of iron, aluminium, calcium, magnesium and beryllium. The red-mud waste contains beryllium at levels above the permissible limit, which prevents its disposal as solid waste. The total beryllium content in the red-mud analysed showed value ranging from 0.39 to 0.59% Be The studies showed that 50% of the total beryllium in red mud can be extracted by water by repeated leaching over a period of 45 days. The cement mix, casting into cement blocks, was subjected to leachability studies over a period of 105 days and immobilization factor (IF factor) was determined. These IF values, of the order of 102, were compared with those obtained by performing leachability study on vitrified red-mud masses produced at different temperature conditions. Direct heating of the red-mud gave the gray coloured, non transparent vitreous mass (as 'bad glass') showed effective immobilisation factor for beryllium in red-mud of the order of 10(4). PMID- 12092764 TI - Agricultural solid waste for the removal of organics: adsorption of phenol from water and wastewater by palm seed coat activated carbon. AB - Adsorption studies for phenol removal from aqueous solution on activated palm seed coat carbon (PSCC) were carried out under varying experimental conditions of contact time, phenol concentration, adsorbent dose and pH. Adsorption equilibrium was reached within 3 h for phenolic concentrations 10-60 mg l(-1). Kinetics of adsorption obeyed a first order rate equation. The percent removal remained constant over the pH range 4-9 for a phenolic concentration of 25 mg (l-1). The equilibrium data could be described well by the Freundlich isotherm equation. The adsorption of phenol on PSCC follows the film diffusion process. A comparative study with a commercial activated carbon showed that PSCC is two times more effective than commercial activated carbon. The studies showed that the palm seed coat carbon can be used as an efficient adsorbent material for the removal of phenolics from water and wastewater. PMID- 12092766 TI - The practice and challenges of solid waste management in Singapore. AB - This paper presents an overview of the current solid waste management situation in Singapore and provides a brief discussion of the future challenges. Singapore is a small island city-state with a large population, warm climate and high humidity. Over the past two to three decades, rapid industrialization and economic development have caused a tremendous increase in solid waste generation. The yearly disposed solid waste increased from 0.74 million tonnes in 1972 to 2.80 million tonnes in 2000. Solid waste management in Singapore has traditionally been undertaken by the Ministry of Environment (ENV), with the participation of some private sectors in recent years. The hierarchy of solid waste management in Singapore is waste minimization (reduce, reuse and recycle or so-called 3 Rs), followed by incineration and landfill. As land is extremely scarce and only one newly constructed offshore landfill site is available, solid waste incineration has been identified as the most preferred disposal method. Waste minimization, the utilization of incineration ashes, industrial waste management are regarded to be the major challenges in the future. PMID- 12092767 TI - Grading of chronic synovitis--a histopathological grading system for molecular and diagnostic pathology. AB - The following is a proposition for a simple histopathological classification system to measure inflammation in synovial tissue. This synovitis-score is employed in conventionally stained routine sections, and is applicable to every kind of synovitis, irrespective of etiology and including the following relevant morphological alterations. First: hyperplasia/enlargement of synovial lining cell layer. Second: activation of resident cells/synovial stroma. Third: inflammatory infiltration. All defined histopathological qualities are graded from absent (0), slight (1) and moderate (2) to strong (3), with summaries ranging from 0 to 9. 0 to 1 corresponds to no synovitis (inflammatory grade = 0), 2 to 3 to a slight synovitis (inflammatory grade 1), 4 to 6 to a moderate synovitis (inflammatory grade 2), and 7 to 9 to a strong synovitis (inflammatory grade 3). Using this score, we analyzed 308 random specimens of synovial tissue from degenerative (osteoarthritis (OA)) and inflammatory joint diseases - reactive arthritis (ReA), psoriasis arthritis (PA) and rheumatoid arthritis (RA) - as well as synovial tissue from healthy individuals. The mean grade given to synovitis of RA turned out to be significantly higher than the mean grade of OA (p < 0.0005) and of healthy controls (p < 0.0005). On the contrary, no significant differences could be found between the mean grades of synovitis scores from patients with RA and those with PA and ReA. Another comparison between RA-synovitis types I and II according to the Stiehl classification resulted in type I (p < 0.0005), showing significantly higher values of inflammatory infiltration, and type II (p = 0.037), showing significantly higher values of stroma activation. Since in OA, synovitis is regarded as a result of degenerative cartilage destruction whereas in inflammatory joint diseases (RA, PA, ReA), synovitis is regarded to be the cause of cartilage destruction, it can be concluded that scores with considerable high values indicate the pathogenetic potential of synovitis and that the inflammatory score may be helpful in estimating the destructive potential of synovitis at the same time. Furthermore, the comparison of the score data with the Stiehl RA-synovitis types shows that the score enables us to discriminate the morphological peculiarities of the synovitis types. In experimental pathology, it could provide standardized information on molecular synovial tissue analyses where a correlation of molecular with morphological data is essential. In diagnostic pathology, this synovitis score (in combination with other typing systems) could provide basic and standardized information concerning the degree of inflammatory alterations in synovial tissue. PMID- 12092768 TI - Epstein Barr virus-associated lymphoma in HIV-infected children. AB - The relationship between Epstein Barr Virus (EBV) and the human host is commonly benign, whereas the development of malignancy is most likely due to imbalance between the virus and host's immune system. The aim of this study was to analyze the association of EBV with pediatric lymphomas in human immunodeficiency virus (HIV) patients. Four consecutive patients with a histological and clinical diagnosis of lymphomas among 351 pediatric HIV-infected children prospectively followed up at our hospital since 1991 were studied. The cases included one diffuse fibrosis lymphocyte depletion subtype Hodgkin's lymphoma, 2 Burkitt's lymphomas, and one primary diffuse large B-cell lymphoma of the central nervous system. We assessed EBV presence by LMP-1 protein labeling by immunohistochemistry and in situ hybridization for EBERs in formalin-fixed and paraffin-embedded biopsies from all four cases. All HIV-associated lymphomas studied were found to be associated with EBV. The lymphoproliferative action of EBV may induce oncogenesis by increasing the probability of genetic alterations and/or by expanding an already malignant clone. As an oncogenic protein, LMP-1 expression by tumor cells supports the involvement of EBV in disease pathogenesis. PMID- 12092769 TI - Decreased expression of TGF-beta type 2 receptor in primary B-cell lymphomas of the stomach. AB - TGF-beta insensitivity has been reported in some malignant lymphomas showing loss of TGF-beta receptor expression. This loss of TGF-beta sensitivity is thought to have removed the immunosuppressive properties of TGF-beta, thus enhancing cell proliferation and resulting in the development of malignant lymphoma. In this study, we performed immunohistochemical stains for TGF-beta1, TGF-beta RI and TGF beta RII in primary gastric B-cell lymphomas in order to ascertain their possible roles in lymphomagenesis. A total of twenty cases of gastric lymphoma were included. All cases of low- and high-grade lymphomas were negative or weakly positive for TGF-beta1. Reactive lymphoid cells, including the germinal center, were also negative for TGF-beta1. In contrast, reactive germinal centers showed moderate to strong cytoplasmic or membranous staining for TGF-beta RI and TGF beta RII. In malignant lymphomas, TGF-beta RI expression was maintained in all cases of low- and high-grade lymphomas. In contrast, TGF-beta RII expression was decreased in all low- and high-grade lymphoma cells. These findings suggest that the loss of TGF-beta RII expression may be involved in the lymphomagenesis of the stomach. PMID- 12092770 TI - Autopsy-proven determinants of death in HIV-infected patients treated for pulmonary tuberculosis in Sao Paulo, Brazil. AB - The aim of this work is to describe and compare pulmonary pathology and proven causes of death in HIV-infected and non-HIV patients treated for tuberculosis, to identify the presence and extension of the lesions, and to suggest appropriate interventions based on the results. Of 246 adult patients (aged > 15) autopsied with tuberculosis and tested for HIV infection at Hospital das Clinicas, School of Medicine, Sao Paulo University, from January 1994 to December 1996, 100 HIV infected patients were matched with 44 non-HIV-infected patients. Demographic determinants influencing patients' death were as follows: 1. Age and sex were not found to be important for the histological outcome, but do seem to correlate with HIV infection. Older patients with tuberculosis are less likely to be HIV infected; 2. Previous tuberculosis and its treatment had no influence on the course of secondary or reinfection tuberculosis; 3. The efficiency of the diagnostic criteria used at the time of death was very low (59.2%); 4. Tuberculosis was more frequently investigated in HIV-infected than in non-HIV infected patients; 5. Only 79 (56%) of the patients received first line agents for treatment; 6. Patients presented with advanced disease; 7. Their mortality is high, and death occurs early. Morphological determinants with influence on patient's death were related to differences in the spectrum of tuberculosis presentation and time of treatment. Noncaseating generalized multibacillary tuberculosis was likely to be the primary cause of death in HIV-infected patients who died during therapy, whereas deaths of patients occurring after the second course of treatment because of recurrence or incomplete treatment were increased for other manifestations of HIV disease (pyogenic pneumonia, Pneumocystis carinii pneumonia, cerebral toxoplasmosis, wasting syndrome). In these patients, dimorphic tuberculosis, an intermediate reactive spectrum form of presentation, was the predominant histological finding. In the opposite spectrum, paucibacillary tuberculosis, a reactive form of tuberculosis presentation, was equally regarded in non-HIV-infected patients as the primary cause of death after four months of therapy. In the same spectrum, non-HIV-infected patients with tuberculosis, who completed all or most of their treatment, died of associated diseases after therapy (alcoholism, cancer, diabetes mellitus). Autopsy-proven determinants of death were associated with HIV status, the spectrum of tuberculosis presentation, and time of treatment. Early treatment for tuberculosis and associated diseases can improve survival and the quality of life even of highly immunosuppressed patients. PMID- 12092771 TI - Accuracy of high resolution CT in assessing idiopathic pulmonary fibrosis histology by objective morphometric index. AB - To determine the accuracy of HRCT in assessing histology by objective morphometric index, twenty-five biopsy specimen-proved UIP were correlated with high-resolution CT (HRCT) by morphometric analysis. The scans were evaluated for the presence and extent of normal parenchyma, ground-glass attenuation, linear opacities, consolidation, honeycombing, vessels and bronchiectasis, and overall extent of histology involvement for normal parenchyma, honeycombing, alveolar septal inflammation, fibrosis, vessels, and bronchiectasis/bronchiolectasis. The comparison between morphometric measurements showed a strong correlation between HRCT and histologic parameters for extension (%) of normal tissue (p = 5 x 10( 5)), honeycombing (p = 6 x 10(-5)), and vessels (p = 0.0047). HRCT consolidation strongly correlated with alveolar septal inflammation (p = 0.015), whereas HRCT linear opacities had the highest correlation with histology for bronchiectasis or bronchiolectasis (p = 0.03). These associations also demonstrated that there was considerable residual scatter about the linear relationships found. By contrast, neither the ground glass patterns nor the bronchioectatic patterns determined by CT were associated with any histologic observation (p < 0.1). There was a borderline negative relationship between vessels determined by CT and histologic fibrosis (p = 0.069), i.e., the percentage of vessel patterns determined by CT was found to be lower when fibrosis was prominent histologically. Our results showed that HRCT patterns, usually employed to provide information about activity (ground glass) and fibrosis (consolidation) in IPF, failed to correlate with histology. On the other hand, chronic cystic lesions had a good correlation with histology. This finding suggests that in patients without a diffuse honeycomb pattern on HRCT, a lung biopsy may provide additional information. The more important limitation of our study was the lack of correlation related to the proximity of the biopsy site to the HRCT location evaluated by morphometry. PMID- 12092772 TI - Acute remodeling of parenchyma in pulmonary and extrapulmonary ARDS. An autopsy study of collagen-elastic system fibers. AB - This study aimed at evaluating acute pulmonary remodeling, focusing on alterations of fibers of the collagenous and elastic systems in ARDS in the exudative phase according to the etiology of the disease. ARDS patients (n = 23), who died in our institution between 1989 and 1997, were retrospectively studied. Ten patients who died in accidents, without any pathological changes in the lung, and ten patients with Congestive Heart Failure (CHF), submitted to mechanical ventilation, were used as control groups. Histological slides were sampled from the autopsied lungs and stained by the Picrosirius and Weigert's resorcin-fuchsin methods. The fiber content of the collagenous and elastic systems of the alveolar septum was measured by image analysis. All patients were in the early ARDS phase (n = 23), 10 pulmonary and 13 extra-pulmonary diseases. Collagen content was greater in pulmonary (1.23+/-0.27) than in extra-pulmonary (0.92+/-0.39) ARDS in the early phase of the disease (p = 0.05). No differences were observed concerning the elastic fibers' content. Extracellular matrix (ECM) remodeling occurs early in the development of acute lung injury and appears to depend on the site of initial insult (pulmonary or extrapulmonary). The present study provides the basis for a prospective, controlled investigation. PMID- 12092773 TI - Angiomyolipoma of the liver and lung: a case explained by the presence of perivascular epithelioid cells. AB - We report a case of synchronous hepatic and pulmonary angiomyolipoma not associated with tuberous sclerosis or renal angiomyolipoma. The liver tumor contained tortuous vessels, smooth muscle tissue, and fat. It was partially necrotic and made up of pleomorphic epithelioid smooth muscle cells. Positivity for HMB-45 confirmed the diagnosis of angiomyolipoma. Lung biopsy showed multiple abnormal proliferation of smooth muscle cells exhibiting spindle-shaped or epithelioid morphology. The tumor grew around the vessels, and the cells were positive for HMB-45. The occurrence of this case could be explained by a simultaneous proliferation of perivascular epithelioid cells. To the best of our knowledge, this is the first case of hepatic angiomyolipoma associated with multiple pulmonary angiomyolipomas, mimicking hepatic tumor lung metastases on X ray examination. PMID- 12092774 TI - Primary bony peripheral T-cell lymphoma mimicking nasal type NK/T-cell lymphoma: a case report. AB - Primary bony lymphomas are rare, and nearly all are high-grade B-cell lymphomas. Natural killer (NK)/T-cell lymphomas are highly aggressive lymphomas of NK- or T cell lineage with predominant extranodal presentation and are divided into nasal and nasal-type (extra-nasal). We report a primary bony peripheral T-cell lymphoma mimicking NK/T-cell lymphoma, nasal type. A 22-year-old Taiwanese male presented with a frontal skull bone mass noted for 3 weeks, and received craniectomy with tumor removal. His tumor showed extensive coagulative necrosis with angioinvasion by large lymphoma cells expressing CD2, CD8, CD16, CD43, CD45, CD45RO, CD56, T cell intracellular antigen-1, and granzyme B, but not CD3, CD4, CD20, CD57, CD68, and betaF1. In situ hybridization for Epstein-Barr virus-encoded mRNA was negative. Polymerase chain reaction study of formalin-fixed tissue showed clonal rearrangement of the T-cell receptor-gamma chain gene. The diagnosis was peripheral T-cell lymphoma, unspecified subtype. The initial stage was I(EA). His lymphoma was refractory to chemotherapy, and bony metastases developed in the right iliac bone 2 months later. He died of disease after 6 months without autopsy. We emphasize the importance of detailed immunohistochemical and gene rearrangement studies for the classification of malignant lymphomas via a very rare primary bony lymphoma of peripheral T-cell subtype. PMID- 12092775 TI - A case of idiopathic nodular glomerulosclerosis mimicking diabetic glomerulosclerosis (Kimmelstiel-Wilson type). AB - A case of idiopathic nodular glomerulosclerosis mimicking diabetic Kimmelstiel Wilson glomerulopathy is reported. The patient was a 45-year-old man suffering from nephrotic syndrome. Light and electron microscopy revealed diffuse and nodular glomerulosclerosis indistinguishable from diabetic nodular glomerulosclerosis. Diabetes mellitus, however, had been excluded both by extensive clinical and by laboratory investigation. The differential diagnosis also included amyloidotic and non-amyloidotic fibrillary glomerulopathy, light chain glomerulopathy, collagen type III disease, immunotactoid glomerulopathy, and the sclerosing variant of membranoproliferative glomerulonephritis. Immunohistochemistry and ultrastructural investigations, however, excluded these entities, and the diagnosis of idiopathic nodular glomerulosclerosis was made. This variant has to be included in the differential diagnosis of nodular glomerulopathy associated with nephrotic syndrome. PMID- 12092776 TI - Seasonal abundance of adult ixodid ticks infesting cattle belonging to resource limited farmers in the north-eastern Free State Province of South Africa. AB - A study was undertaken to determine the seasonal abundance of the ticks infesting cattle owned by resource-limited farmers in the north-eastern Free State Province of South Africa. Infestations of Boophilus decoloratus Koch, 1844, Rhipicephalus evertsi evertsi Neumann, 1897, Rhipicephalus follis Donitz, 1910, Rhipicephalus gertrudae Feldman-Muhsam, 1960 and Rhipicephalus warburtoni Walker & Horak, 2000, were monitored on cattle of mixed breeds at monthly intervals from May 1998 to April 1999. High tick intensity on the cattle was observed between March and June, with a peak in May to June 1998 for B. decoloratus and R. evertsi evertsi. Few ticks from the other three species were recovered from the cattle. Small peaks in November to December 1998 for R. warburtoni, and in December 1998 for R. follis and R. gertrudae were observed. Following the winter (June to August), numbers of B. decoloratus declined markedly, while those of R. evertsi evertsi and other rhipicephalids increased. These results represent the first published information on ticks of veterinary importance infesting cattle in the north eastern part of the Free State Province. PMID- 12092777 TI - Redescription of some Thelandros and Tachygonetria spp. (Pharyngodonidae: Oxyuroidea) from the omnivorous plated lizard, Gerrhosaurus validus validus A. Smith, 1849 in South Africa. AB - Thelandros schusteri Hering-Hagenbeck, 2001, Thelandros luciusi Hering-Hagenbeck, 2001, Thelandros boomkeri Hering-Hagenbeck, 2001, Tachygonetria bainae Hering Hagenbeck, 2001, Tachygonetria chabaudi Hering-Hagenbeck, 2001 and Tachygonetria petterae Hering-Hagenbeck, 2001 from the plated lizard, Gerrhosaurus validus validus A. Smith 1849 from three localities in the north-eastern region of South Africa are redescribed. Classification keys are available only for the males of the species and because male and female nematodes in copula were not observed in this study as well as the similarity of the females, it was not possible to identify the females to the species level. Thelandros schusteri, Thelandros boomkeri and Thelandros luciusi were provisionally paired with female Type E, Tachygonetria bainae with female Type C, Tachygonetria chabaudi with female Type A and Tachygonetria petterae with female Type D. Female Types B and F could not be paired. The richness and composition of species of the Pharyngodonidae of Gerrhosaurus validus validus is close to that of tortoises and differs from the pharyngodonid fauna of the insectivorous lizards that have been studied. In the latter, only the genera Spauligodon, Skrjabinodon and Parapharyngodon were recovered. The pharyngodonid fauna of Gerrhosaurus validus validus seems to have originated by capture from local herbivorous reptiles. The three Tachygonetria spp. most closely resemble forms in South African tortoises. The three Thelandros spp. redescribed here not only show strong similarities to those of herbivorous Agama spp., but also to those parasitic in tortoises and could have been acquired from either. PMID- 12092778 TI - Description of Pelonia africana n. g., n. sp. (Pentastomida: Sebekidae) from the lungs of Pelomedusa subrufa and Pelusios sinuatus (Chelonia) in South Africa. AB - The terrapins Pelomedusa subrufa and Pelusios sinuatus taken from a water reservoir in the Northern Province, South Africa, were examined for pentastome infections. Two immature specimens, a patent female and a mature male, were obtained from the lungs of four hosts, each of which harboured a single specimen. Based on the morphology of the specimens the new monospecific genus, Pelonia africana n. g., n. sp., is described. It is characterized by smooth, dorsally convex hooks with sharply bent blades. The oral cadre is more or less U-shaped. Delicate chitinous fibres, which can be difficult to see, connect the lateral prongs anteriorly. In this, as well as the morphology of the copulatory spicules, it is most like Sebekia wedli. The latter, however, possesses spinous hooks, which are absent in P. africana. The hooks are slightly and the copulatory spicules markedly larger in P. africana than in S. wedli. The lack of a double hooked collar at the terminal end of the cowry-shell shaped base of each copulatory spicule and the absence of a peg-like extension of the oral cadre into the oesophagus, distinguishes P. africana from members of the genus Alofia. The oral cadre of the South American species Diesingia megastoma, from aquatic chelonians, is more than twice the size than that of Pelonia and there is a distinct difference in shape. The hooks of the genus Diesingia are flat-topped, and both the anterior as well as the posterior fulcra carry cowl-like extensions. The number of annuli, 55-60 in D. megastoma and approximately 30 in P. africana, further separates the two genera. The most striking feature of Diesingia which sets it apart from Pelonia and the other genera of the family Sebekidae, is the configuration of its copulatory spicules. Pelonia and Diesingia share morphological features with all the other sebekiids, but it is the unique combination of diagnostic characters that separates the two genera from those, as well as from each other. PMID- 12092779 TI - Validation of an indirect enzyme-linked immunosorbent assay for the detection of antibody against Brucella abortus in cattle sera using an automated ELISA workstation. AB - An automated indirect enzyme-linked immunosorbent assay (I-ELISA) for the serological diagnosis of bovine brucellosis was developed and validated in-house. A total of 4,803 cattle sera from South Africa (n = 3,643), Canada (n = 652), Germany (n = 240), France (n = 73) and the USA (n = 195) was used. The South African panel of sera represented 834 sera known to be positive by the Rose Bengal test (RBT), serum agglutination test (SAT) and complement fixation test (CFT), 2709 sera that were negative by CFT, and 100 sera from animals vaccinated with a standard dose of Brucella abortus strain 19. Overseas sera were obtained from reference non-vaccinated brucella-free cattle (n = 834), naturally infected (n = 72), experimentally infected (n = 71), and vaccinated animals (n = 83). Also 100 sera collected from cattle in Canada and known to be positive by competitive ELISA (C-ELISA) were used. The intermediate ranges ("borderline" range for the interpretation of test results) were derived from two-graph receiver operating characteristics analysis. The lowest values of the misclassification cost-term analysis obtained from testing overseas panels, covered lower I-ELISA cut-off PP values (0.02-3.0) than those from local panels (1.5-5.0). The relatively low cut off PP values selected for I-ELISA were due to the fact that the positive control used represents a very strong standard compared to other reference positive sera. The greater overlap found between negative and positive cattle sera from South Africa than that between reference overseas panels was probably due to the different criteria used in classifying these panels as negative (sera from true non-diseased/non-infected animals) or positive (sera from true diseased/infected animals). The diagnostic sensitivity of the I-ELISA (at the optimum cut-off value) was 100% and of the CFT 83.3%. The diagnostic specificity of I-ELISA was 99.8% and of the CFT 100%. Estimate of Youden's index was higher for the I-ELISA (0.998) than that for the CFT (0.833). Analysis of distribution of PP values in sera from vaccinated and naturally infected cattle shows that in vaccinated animals all readings were below 31 PP where in infected ones these values represented 43%. Therefore, it appears that I-ELISA could be of use in identifying some naturally infected animals (with values > 31 PP), but more sera from reference vaccinated and infected animals need to be tested to further substantiate this statistically. Of 834 sera positive by RBT, SAT and CFT, 825 (98.9%) were positive in the I-ELISA. Compared to C-ELISA the relative diagnostic sensitivity of the I-ELISA was 94% and of the CFT 88% when testing 100 Canadian cattle sera. Of 258 South African cattle sera, of which 183 (70.9 %) were positive by the I-ELISA and 148 (57.4 %) by the CFT, 197 (76.4%) were positive by C-ELISA when re-tested in Canada. One has to stress, however, that Canadian C ELISA has not been optimised locally. Thus, the C-ELISA was probably not used at the best diagnostic threshold for testing South African cattle sera. This study shows that the I-ELISA performed on an automated ELISA workstation provides a rapid, simple, highly sensitive and specific diagnostic system for large-scale detection of antibodies against B. abortus. Based on the diagnostic accuracy of this assay reported here, the authors suggest that it could replace not only the currently used confirmatory CFT test, but also the two in-use screening tests, namely the RBT and SAT. PMID- 12092780 TI - Redescription of some Spauligodon spp. and Parapharyngodon spp., and of skrjabinodon mabuyae (Sandground, 1936) inglis, 1968 (Pharyngodonidae: Oxyuroidea) from insectivorous South African lizards. AB - As part of a study on the helminth parasites of South African lizards several species of saurians were collected from localities in the North West Province, the Northern Province, Mpumalanga Province and Gauteng Province. Spauligodon blydeensis (Hering-Hagenbeck, 2001) from the Cape thick-toed gecko, Pachydactylus capensis, Spauligodon molpoensis (Hering-Hagenbeck, 2001) from Wahlberg's velvet gecko, Homopholis wahibergii, Parapharyngodon margaritiferi, Hering-Hagenbeck, 2001 from the skink, Mabuya margaritifer, Parapharyngodon gerrhosauri, Hering Hagenbeck, 2001 from the plated lizard, Gerrhosaurus flavigularis and Skrjabinodon mabuyae (Sandground, 1936) Inglis, 1968 from the skinks Mabuya punctatissima, Mabuya spilogaster and Mabuya varia are redescribed. The different Spauligodon spp. in the subcontnent may be separated on the presence or absence of spicules in the males, the presence or absence of spines on the tail of both the males and females, as well as on the size and shape of the eggs, and the configuration of the polar caps. The Parapharyngodon spp. are distinguished mainly by the morphological characters of the males, such as the width of the caudal alae and the size of the pre- and adanal papillae. Female Parapharyngodon spp. closely resemble each other and some could not be identified to the species level since males were absent. Spinose larvae, together with adult Parapharyngodon spp. were recovered from Mabuya margaritifer. All Parapharyngodon spp. larvae described to date are spinose and since the larvae in this study were collected together with adult Parapharyngodon spp., we consider them to belong to the same genus. Skrjabinodon mabuyae differs from the closely related Skrjabinodon mabuiensis in the presence of a spicule in the male and lateral alae in the female. The former nematode is described for the first time from skinks in South Africa. PMID- 12092782 TI - Teratogenicity of a mutagenised Rift Valley fever virus (MVP 12) in sheep. AB - A 5-fluorouracil mutagenised Rift Valley fever virus strain, which was shown to be attenuated and immunogenic in cattle and sheep, was evaluated for its ability to cause teratogenic effects in pregnant sheep. A group of 50 sheep at various stages of pregnancy was inoculated with the virus and the pregnancies followed to term. There were two abortions and 14% of the lambs produced by vaccinated ewes showed teratogenic effects, the most prevalent being spinal hypoplasia, hydranencephaly, brachygnathia inferior and arthrygryposis. The foetal malformations of the central nervous and musculo-skeletal systems were mostly consistent with those observed in sheep vaccinated with the attenuated Smithburn RVF strain. The teratogenic effects of MVP12 were not seen in previous experiments by other authors as immunisation of sheep took place in the second to third trimester of pregnancy, when the foetal brain tissue has completed most of its cell division. PMID- 12092781 TI - The classification of seven serotypes of equine encephalosis virus and the prevalence of homologous antibody in horses in South Africa. AB - Selected isolates of equine encephalosis virus were shown to have comparable viral protein profiles and to represent seven distinct serotypes, based on cross neutralization tests. Serotype-specific virus-neutralizing antibody in serum samples from horses confirmed the widespread occurrence of infection. The distribution and prevalence of individual serotypes however, varied considerably. Localised foci with an increased seasonal seroconversion in groups of horses to a specific serotype and the detection of an ongoing low level of infection from other serotypes within the population, confirmed the independent persistence of the viruses in a maintenance cycle. The identification of donors with antibody resulting from infection with multiple serotypes indicated a low level of cross protection in horses to natural reinfection. PMID- 12092783 TI - Cardiovascular characteristics and cerebral CT findings in elderly subjects with psychomotor disadaptation syndrome. AB - BACKGROUND AND AIMS: In previous studies, we described a clinical picture typically observed in frail old people, called "Psychomotor Disadaptation Syndrome" (PDS), but we have never studied etiopathogenic data. The aim of this study was to complete the clinical picture, record cardiovascular risk factors and provide cerebral CT scan findings in patients with PDS. METHODS: 73 patients with PDS were recruited in the Geriatric Medicine departments of the University Hospitals of Dijon (Burgundy, France) and Strasbourg (Alsace, France); this group included prospectively all hospitalized patients showing postural and gait abnormalities according to the following criteria: trend towards backward falling and gait pattern alteration characterized by hesitancy in initiation, small steps, and increase in the double support durations. General characteristics, neurological and cardiovascular information were collected for each patient. For the detection of white matter changes (WMC), we used a third-generation CT scanner (GE CT HSA) evaluating a section of 7 mm at each interval of 8 mm. RESULTS: Neurological examination showed that "reactional hypertonia" was observed in more than 90% of the patients, and that no patient showed normal reactive postural responses. Prior history of hypertension was noted in 49% of the patients, while a current antihypertensive treatment was taken by 13% of the patients. Orthostatic hypotension was observed in 44% of patients. Severe or moderate periventricular lucencies on CT scan were found in 67% of the patients, and severe ventricular enlargement in 50.5% of the patients. CONCLUSIONS: We advance that PDS might be associated with WMC, and that hypotension might be an important etiologic factor of WMC in causing reduction of the cerebral blood flow in subcortical areas. PMID- 12092784 TI - Cerebral perfusion deficits in age-associated memory impairment. The role of tobacco smoking. AB - BACKGROUND AND AIMS: Several studies report a high prevalence of memory complaints in the elderly, non-demented population. Since the pathogenesis and diversity of memory impairment in healthy people is still an unresolved issue, we designed our study to ascertain the occurrence of brain abnormalities in Age Associated Memory Impairment (AAMI) by measuring regional cerebral blood flow (rCBF). METHODS: Fifty-three subjects were invited to the case-control study: 26 subjects who fulfilled the AAMI criteria, and 27 controls (age, mean+/-SD 65.8+/ 7.3 vs 65.5+/-6.7). rCBF was determined using 99mTC-ECD SPECT by relative measurements made in the cortical and white matter regions and in the thalamus. RESULTS: The AAMI group vs controls had restricted rCBF in the temporal lobe of the left hemisphere (median; 85.8 vs 87.8, p=0.04), in the thalamus (median; 87.3 vs 89.8, p=0.036) and the frontal white matter (median; right hemisphere 80.0 vs 84.1, p=0.01; left hemisphere 77.7 vs 81.6, p=0.04). AAMI subjects who previously smoked had lower rCBF values in the temporal, parietal, occipital and frontal lobes bilaterally, and poorer memory test results than ex-smoker controls. By contrast, rCBF did not differ between non-smoking AAMI subjects and controls. CONCLUSIONS: Compared to controls, AAMI subjects had restricted rCBF, A history of previous smoking could partly contribute to the rCBF differences between these groups. Further studies on failing memory and rCBF in representative cohorts of smoking and non-smoking populations are now needed. PMID- 12092785 TI - Variability of trail making test, symbol digit test and line trait test in normal people. A normative study taking into account age-dependent decline and sociobiological variables. AB - BACKGROUND AND AIMS: The influence of sociobiological variables and aging on the variability of the Trail Making Tests (TMT), the Symbol Digit Substituting Test (SDT), and the Line Trait Test (LTT) in the general healthy populations are not well known. Even less is known about the reliability at re-testing. This study aimed at determining the reference range of these tests, taking into account sociobiological variables and age, and the re-testing effect. METHODS: We studied 300 healthy subjects from 20 to 80 years of age. The sample was derived by the pooling of two samples stratified by age and sex: a randomized sample of 161 subjects collected from the city registers of Padova, and a convenience sample of 139 subjects collected in 20 towns (mainly rural) of Northern Italy. After normalization, data were assayed for the influence of age, education, job, and gender. RESULTS: Age was found to be a significant independent predictor for all the tests, education for all but the LTT, job only for the TMT-B and a geometrical version of the same test (TMT-G) which was proved to be highly correlated with the TMT-B (r=0.80, p<0.01). Job and the interaction age x education level influenced the difference TMT-B minus TMT-A. From the predicting equations, the normative data and the formulas to obtain Z scores for each test were derived. Reliability was lowest for LTT errors (CV=67%), highest for the SDT (13%), whereas the TMT obtained intermediate values (22-33%, depending on the test). CONCLUSIONS: This study provides the most reliable normative data range for the TMT, SDT and LTT to date because it considers important demographic variables such as age, education and job. PMID- 12092786 TI - Cognitive decline is related to education and occupation in a Spanish elderly cohort. AB - BACKGROUND AND AIMS: The type of education and occupation can sensibly influence cognitive decline. The aim of this study was to examine their impact on cognitive function in a longitudinal study of community-dwelling Spanish people over 65 with low levels of formal education and predominantly unskilled occupations. METHODS: Cognitive function was assessed in 1993 and 1997 using a simple scale including items of memory and orientation that has been previously validated for populations with low levels of formal education. Cognitive score in 1997 and cognitive decline over 4 years (1993-1997) were used as outcomes. Education and occupation were analyzed as determinants of cognitive function using multiple linear regression, and of cognitive decline using logistic and multinomial regressions. RESULTS: Of the 557 subjects who completed the follow-up in 1997, 11% had experienced severe decline and 20.6% mild decline. Overall and mild cognitive decline were predicted by low education and being a farm worker (OR: 2.36, CI 95%: 1.164.81 and OR: 2.37, CI 95%: 1.05-5.37) after controlling for age. CONCLUSIONS: Cognitive decline in the elderly is partially explained by early life events, such as education, and living in a deprived environment over a long period of time. We cannot ascertain whether these effects are direct or mediated by other associated conditions but sample attrition does not account for our results. PMID- 12092787 TI - Mortality in three different cohorts of 70-year olds: the impact of social factors and health. AB - BACKGROUND AND AIMS: Little is known about changes in mortality among elderly over time, especially in subgroups differing in social and other background factors. The aim of this study was to analyze differences in mortality in three different cohorts of 70-year olds regarding social factors, social network, self assessed health and number of diseases. METHODS: We used data from 3 random samples of 70-year olds born in 1901/02 (N=973), 1906/07 (N=1036) and 1911/12 (N=619). At age 70 all participants were given medical examinations and interviewed regarding social background, social network, self-assessed health, need of care and number of diseases. Death records were obtained up to and including 1998. RESULTS: The youngest cohorts had significantly decreased mortality compared to the oldest cohort (RR 0.86; CI 0.78-0.95 and RR 0.88; CI 0.78-0.995 respectively). There were no differences between the two youngest cohorts. Decrease in mortality in the younger cohorts was found among those who lived in their own home, did not feel tired, were non- or ex-smokers and had one or more diseases. CONCLUSIONS: There was a small difference in total mortality between these cohorts of 70-year olds. There were differences in development of mortality in subgroups distributed by health and social factors, indicating more years of life, partly due to life-style factors such as non-smoking. PMID- 12092788 TI - Low level dietary restriction retards age-related dolichol accumulation. PMID- 12092789 TI - Living longer--but better? AB - The highest attained age has increased by about 20 years since the beginning of the 19th century. In the course of the 1990s, more than ten individuals reached 115 years or more, including Jeanne Calment who attained the age of 122 years. In low-mortality countries, the number of centenarians has doubled every decade since 1950. This dramatic increase was mainly due to periodical effects related to the drastic fall in mortality among the elderly. The fact that centenarians are survivors does not mean that they are healthy. A high prevalence of comorbidity is found, and many centenarians have survived major diseases thanks to medical treatment and surgery. It is, however, possible that the comorbidity is less serious than in younger elderly. Certain personality traits may also be important in surviving health-threatening conditions. Furthermore, a number of biological and cognitive functions seem to be well-preserved in several centenarians. The influence of the apoE-gene and other genes involved in fundamental mechanisms illustrates that with advancing age and increasing mortality even small risks may have a substantial effect on survival to 100 years. A small proportion of long-livers may be considered as relatively autonomous, and this proportion will probably increase in the future. We are living longer and seem to postpone the terminal dependent phase to higher ages. Longevity may thus be perceived as part of our postmodern condition with its mix of pleasure and suffering. PMID- 12092791 TI - Erasmus Darwin (1731-1802). PMID- 12092790 TI - Circadian temporal organization of lipidic fractions in elderly people. Entrainment to the dietary schedule. AB - BACKGROUND AND AIMS: Changes in some rhythmometric parameters have been reported in the elderly as a consequence of both structural and neurochemical changes occurring in the central nervous system. Since alterations of lipid and lipoprotein metabolism are directly involved in several age-related disorders, the aim of this study was to investigate the circadian temporal organization of some important lipidic fractions (total cholesterol, triacylglycerol, apolipoprotein A1 and B) in physiological aging. METHODS: Thirty old hospitalized subjects were synchronized for daily activities, sleeping/waking habits, and time/quality of meals. Twenty-four healthy young individuals served as controls. After an overnight fast, samples were taken beginning at 08:00 every 4 hours until 20:00, and every 2 hours from 20:00 to 04:00. Rhythmometric data were analyzed by single and population mean Cosinor analysis, and by ANOVA; the comparison of the rhythm's parameters between elderly and young subjects was carried out by the Mesor test and the amplitude-acrophase using Hotelling's test. RESULTS: Elderly subjects exhibited statistically significant circadian rhythms for total cholesterol (p<0.00002), triacylglycerol (p<0.000001), apo A-1 (p<0.0013), and apo B (p3)-D-glucan as a potential antitumor agent. AB - Alpha-glucan L-FV-II and beta-glucan L-FV-I were shown to co-exist in the extract of fruiting bodies of Lentinus edodes with aq. 5% NaOH/0.05% NaBH4 in previous work. Water-insoluble alpha-(1-->3)-D-glucan (L-FV-II) was treated with sulfur trioxide-pyridine complex at 25 degrees C to synthesize the water-soluble sulfated derivative SL-FV-II with a degree of substitution (DS) 1.1 in non selective sulfation. The weight-average molecular weight (Mw) of sulfated glucan SL-FV-II is 57% of that of the original alpha-glucan L-FV-II. The alpha-glucan administered by gavaging at a dose of 50 mg/kg of body weight to BALB/C mice having implanted solid Sarcoma 180 was effective at an inhibition rate of 42%. In vitro experiments using human and murine tumor cell lines showed that SL-FV-II had antiproliferation activity at the concentration of 20 microg/mL towards four tumor cell lines. The sulfated alpha-(1-->3)-D-glucan had potent antiproliferation action (52%) on human MCF-7 breast carcinoma cells. PMID- 12092816 TI - Cloning and sequencing of the genes encoding cyclic tetrasaccharide-synthesizing enzymes from Bacillus globisporus C11. AB - The genes for isomaltosyltransferase (CtsY) and 6-glucosyltransferase (CtsZ), involved in synthesis of a cyclic tetrasaccharide from alpha-glucan, have been cloned from the genome of Bacillus globisporus C11. The amino-acid sequence deduced from the ctsY gene is composed of 1093 residues having a signal sequence of 29 residues in its N-terminus. The ctsZ gene encodes a protein consisting of 1284 residues with a signal sequence of 35 residues. Both of the gene products show similarities to alpha-glucosidases belonging to glycoside hydrolase family 31 and conserve two aspartic acids corresponding to the putative catalytic residues of these enzymes. The two genes are linked together, forming ctsYZ. The DNA sequence of 16,515 bp analyzed in this study contains four open reading frames (ORFs) upstream of ctsYZ and one ORF downstream. The first six ORFs, including ctsYZ, form a gene cluster, ctsUVWXYZ. The amino-acid sequences deduced from ctsUV are similar in to a sequence permease and a sugar-binding protein for the sugar transport system from Thermococcus sp. B1001. The third ctsW encodes a protein similar to CtsY, suggested to be another isomaltosyltransferase preferring panose to high-molecular-mass substrates. PMID- 12092817 TI - Phenylethylamine induces an increase in cytosolic Ca2+ in yeast. AB - Beta-phenylethylamine (PEA) induced an increase in cytosolic free calcium ion concentration ([Ca2+]c) in Saccharomyces cerevisiae cells monitored with transgenic aequorin, a Ca2+-dependent photoprotein. The PEA-induced [Ca2+]c increase was dependent on the concentrations of PEA applied, and the Ca2+ mostly originated from an extracellular source. Preceding the Ca2+ influx, H2O2 was generated in the cells by the addition of PEA. Externally added H2O2 also induced a [Ca2+]c increase. These results suggest that PEA induces the [Ca2+]c increase via H2O2 generation. The PEA-induced [Ca2+]c increase occurred in the mid1 mutant with a slightly smaller peak than in the wild-type strain, indicating that Mid1, a stretch-activated nonselective cation channel, may not be mainly involved in the PEA-induced Ca2+ influx. When PEA was applied, the MATa mid1 mutant was rescued from alpha-factor-induced death in a Ca2+-limited medium, suggesting that the PEA-induced [Ca2+]c increase can reinforce calcium signaling in the mating pheromone response pathway. PMID- 12092818 TI - Chitinases A, B, and C1 of Serratia marcescens 2170 produced by recombinant Escherichia coli: enzymatic properties and synergism on chitin degradation. AB - To discover the individual roles of the chitinases from Serratia marcescens 2170, chitinases A, B, and C1 (ChiA, ChiB, and ChiC1) were produced by Escherichia coli and their enzymatic properties as well as synergistic effect on chitin degradation were studied. All three chitinases showed a broad pH optimum and maintained significant chitinolytic activity between pH 4 and 10. ChiA was the most active enzyme toward insoluble chitins, but ChiC1 was the most active toward soluble chitin derivatives among the three chitinases. Although all three chitinases released (GlcNAc)2 almost exclusively from colloidal chitin, ChiB and ChiC1 split (GlcNAc)6 to (GlcNAc)3, while ChiA exclusively generated (GlcNAc)2 and (GlcNAc)4. Clear synergism on the hydrolysis of powdered chitin was observed in the combination between ChiA and either ChiB or ChiC, and the sites attacked by ChiA on the substrate are suggested to be different from those by either ChiB or ChiC1. PMID- 12092819 TI - Functional analysis of the chitin-binding domain of a family 19 chitinase from Streptomyces griseus HUT6037: substrate-binding affinity and cis-dominant increase of antifungal function. AB - Chitinase C (ChiC) is the first bacterial family 19 chitinase discovered in Streptomyces griseus HUT6037. While it shares significant similarity with the plant family 19 chitinases in the catalytic domain, its N-terminal chitin-binding domain (ChBD(ChiC)) differs from those of the plant enzymes. ChBD(ChiC) and the catalytic domain (CatD(ChiC)), as well as intact ChiC, were separately produced in E. coli and purified to homogeneity. Binding experiments and isothermal titration calorimetry assays demonstrated that ChBD(ChiC) binds to insoluble chitin, soluble chitin, cellulose, and N-acetylchitohexaose (roughly in that order). A deletion of ChBD(ChiC) resulted in moderate (about 50%) reduction of the hydrolyzing activity toward insoluble chitin substrates, but most (about 90%) of the antifungal activity against Trichoderma reesei was abolished by this deletion. Thus, this domain appears to contribute more importantly to antifungal properties than to catalytic activities. ChBD(ChiC) itself did not have antifungal activity or a synergistic effect on the antifungal activity of CatD(ChiC) in trans. PMID- 12092820 TI - Short-step syntheses of all stereoisomers of preussin and their bioactivities. AB - All the eight stereoisomers of (+)-preussin (1b), an antifungal agent inhibiting the growth of fission yeast and human cancer cells, were synthesized in two steps by non-stereoselective reactions and chromatographic separation, starting from L- and D-N-protected-phenylalaninal (2). Their bioassay revealed all of the stereoisomers to be almost equally bioactive. PMID- 12092821 TI - Identification, cloning, and sequencing of the genes involved in the conversion of D,L-2-amino-delta2-thiazoline-4-carboxylic acid to L-cysteine in Pseudomonas sp. strain ON-4a. AB - The newly isolated strain Pseudomonas sp. ON-4a converts D,L-2-amino-delta2 thiazoline-4-carboxylic acid to L-cysteine via N-carbamoyl-L-cysteine. A genomic DNA fragment from this strain containing the gene(s) encoding enzymes that convert D,L-2-amino-delta2-thiazoline-4-carboxylic acid into L-cysteine was cloned in Escherichia coli. Transformants expressing cysteine-forming activity were selected by growth of an E. coli mutant defective in the cysB gene. A positive clone, denoted CM1, carrying the plasmid pCM1 with an insert DNA of approximately 3.4 kb was obtained, and the nucleotide sequence of a complementing region was analyzed. Analysis of the sequence found two open reading frames, ORF1 and ORF2, which encoded proteins of 183 and 435 amino acid residues, respectively. E. coli DH5alpha harboring pTrCM1, which was constructed by inserting the subcloned sequence into an expression vector, expressed two proteins of 25 kDa and 45 kDa. From the analyses of crude extracts of E. coli DH5alpha carrying deletion derivatives of pTrCM1 by sodium dodecyl sulfate polyacrylamide gel electrophoresis and by enzymatic activity, it was found that the 25-kDa protein encoded by ORF1 was the enzyme L-2-amino-delta2-thiazoline-4 carboxylic acid hydrolase, which catalyzes the conversion of L-2-amino-delta2 thiazoline-4-carboxylic acid to N-carbamoyl-L-cysteine, and that the 45-kDa protein encoded by ORF2 was the enzyme N-carbamoyl-L-cysteine amidohydrolase, which catalyzes the conversion of N-carbamoyl-L-cysteine to L-cysteine. PMID- 12092822 TI - Analysis of transactivation region of Elf-1 by using a yeast one-hybrid system. AB - The transcriptional regulatory region of Elf-1 was analyzed by the combination of a yeast one-hybrid system and site-directed mutagenesis. This analysis enabled us to map an activation region between 85-175 of Elf-1. PMID- 12092823 TI - Preventive effect of a chicken extract on the development of hypertension in stroke-prone spontaneously hypertensive rats. AB - The antihypertensive effect of Brand's Essence of Chicken (BEC), a popular chicken extract used as a traditional health food, was examined with stroke-prone spontaneously hypertensive rats (SHRSPs). The animals were maintained from 6 to 25 weeks of age on drinking water with or without BEC. The BEC-fed group showed a significant reduction in the development of hypertension when compared with the control animals. The levels of blood urea nitrogen and plasma creatinine in the BEC-fed group were significantly lower than those in the control group, suggesting that the renal glomerular function had been improved by the daily administration of BEC. It thus seems likely that BEC would be useful as a prophylactic treatment against the development of hypertension and renal injury. PMID- 12092825 TI - Characterization of cloned mouse Na+/taurocholate cotransporting polypeptide by transient expression in COS-7 cells. AB - The mouse Na+/taurocholate cotransporting polypeptide transiently expressed in COS-7 cells caused sodium-dependent uptake of [3H]taurocholic acid with Km and Vmax values of 18 microM and 102 pmol/mg protein/min, respectively. This Km value is comparable to that for rat NTCP and higher than that for human NTCP. Substrate specificity was evaluated by measuring inhibitory effects of unlabeled bile acids on [3H]taurocholic acid transport. PMID- 12092824 TI - Selective removal of chymotrypsin using diphenyl alpha-aminoalkylphosphonate immobilized on sepharose gel. AB - A diphenyl alpha-aminoalkylphosphonate derivative, which is an irreversible inhibitor of chymotrypsin-like serine proteases, was immobilized on cyanogen bromide-activated Sepharose, and the selective binding of chymotrypsin to the obtained inhibitor-gel was evaluated using batch and column methods. Complete removal of chymotrypsin in an aqueous solution was done using the column method, while partial removal was done using the batch method. PMID- 12092826 TI - Analysis of hydrolytic activity of a 65-kDa chitinase from the silkworm, Bombyx mori. AB - The hydrolytic reactions of Bombyx mori 65-kDa chitinase with the short substrates, N-acetyl-chitooligosaccharides, were analyzed by HPLC. Analysis of the hydrolyzed products showed that the newly produced oligosaccharides are all beta anomers, suggesting that, similar to other family 18 glycosyl hydrolases, the 65-kDa chitinase acts in the retaining mechanism. Furthermore, the enzyme cleaves the N-acetylchitooligosaccharides mainly at the linkage between the second and the third GlcNAc moieties from the non-reducing end, while the other sites were cleaved in smaller proportions. Moreover, the initial reaction rates of the enzyme with the longer N-acetylchitooligosaccharides were higher than those with shorter ones. These results suggest that the enzyme is an endo cleaving type and more efficient on the longer substrates. PMID- 12092827 TI - Enantioselective hydrolysis of o-nitrostyrene oxide by whole cells of Aspergillus niger CGMCC 0496. AB - The asymmetric biohydrolysis is described of o-nitrostyrene oxide with high selectivity by whole cells of Aspergillus niger CGMCC 0496. Both the epoxide and diol could be obtained in a high state of optical purity (over 98%). Product inhibition was found when using a high ratio of substrate to cells. PMID- 12092828 TI - Relationship between conidial enzymes and germination of the apple blue mold, Penicillium expansum. AB - The relationship between conidial enzymes of Penicillium expansum and spore germination was examined. The activities of xylanase and pectinase, but not of cellulase and amylase, were detected in the conidia. The levels of xylanase and pectinase were greatly enhanced by xylan and pectin as respective carbon sources in the basal medium. No conidia germinated in the basal medium without a carbon source. The type of carbon source and the enzyme levels of the conidia did not affect the rate of germination. However, a relationship was found between the enzyme levels and the elongation of the germ tubes. PMID- 12092829 TI - Release of hemorphin-5 from human hemoglobin by pancreatic elastase. AB - Opioid peptide hemorphin-5 (YPWTQ) was released from human hemoglobin by the action of pancreatic elastase. V-hemorphin-5 (VYPWTQ) was also released under the same conditions. The amount of hemorphin-5 was about 1/10 that of V-hemorphin-5. This is the first report showing release of hemorphin-5 only by the action of one enzyme. PMID- 12092830 TI - Protocol for measuring the endurance capacity of mice in an adjustable-current swimming pool. AB - We re-examined the methods used in an adjustable-current swimming pool for evaluating the endurance capacity of mice to improve the sensitivity and reproducibility. We found that the BALB/c strain was most suitable to minimize the variation in time taken to swim to the point of fatigue. We found that precise adjustment of the apparatus and the use of three primary swimming trials to select mice before a study made the swimming time more uniform. This procedure was used to establish a more precise evaluation protocol. PMID- 12092831 TI - Cloning and sequencing of the serine dehydrogenase gene from Agrobacterium tumefaciens. AB - The structural gene for NADP+-dependent serine dehydrogenase [EC 1.1.1.-] from Agrobacterium tumefaciens ICR 1600 was cloned into Escherichia coli cells and its complete DNA sequence was analyzed. The gene encodes a polypeptide containing 249 amino acid residues. The enzyme had high sequence similarity to short-chain alcohol dehydrogenases from bacteria and unknown proteins of Haemophilus influenzae, Escherichia coli, and Saccharomyces cerevisiae. PMID- 12092832 TI - Cloning and characterization of cDNAs for the jasmonic acid-responsive Genes RRJ1 and RRJ2 in suspension-cultured rice cells. AB - Two cDNA clones for jasmonic acid (JA)-responsive genes, RRJ1 and RRJ2, were isolated by differential screening from suspension-cultured rice cells treated with JA for 2 h. The putative RRJ1 protein is completely identical to that of a putative rice cystathionine gamma-lyase, while the putative RRJ2 protein is highly similar in sequence to a rice pyruvate decarboxylase, PDC1. PMID- 12092833 TI - Functional expression of chitinase and chitosanase, and their effects on morphologies in the yeast Schizosaccharomyces pombe. AB - The chitinase A (chiA) gene from Enterobacter sp. G-1 and the chitosanase A (choA) gene from Matsuebacter chitosanotabidus 3001 were expressed separately and simultaneously in the yeast Schizosaccharomyces pombe. The chiA gene was placed under the transcriptional control of the nmt1 promoter with the glutathione S transferase (GST), and the choA gene was expressed by the human cytomegalovirus (hCMV) promoter with the P factor secretion signal (P3). The expressed proteins of ChoA and GST-ChiA were enzymatical active and were detected as 34-kDa and 80 kDa, respectively, by Western blot analysis. The transformant that expressed the choA gene was able to secrete ChoA efficiently into the culture medium with the specific activity of 102.36 U/mg protein. When the chiA gene was expressed in S. pombe, yeast cells grew slowly and cells became elongated, but when the choA gene was expressed, cells became swollen. Expression of both the chiA and the choA genes resulted in elongated and fat cells. PMID- 12092834 TI - Green fluorescent protein-labeled recombinant fluobody for detecting the picloram herbicide. AB - A green fluorescent protein-labeled fluobody was designed to develop a simple immunoassay method for detecting picloram herbicide in an environmental sample. The gfp gene was successfully inserted into the pSJF2 vector harboring the picloram-specific antibody fragment to yield pSJF2GFP. Picloram spiking in an environmental river sample could be indirectly detected by observing the fluorescence intensity value of the gfp-fluobody, exhibiting specific sensitivity to free picloram with an IC50 value of 50 ppb. Using the gfp-fluobody immunoassay avoids the enzyme-substrate reaction for calorimetric detection that is required in an enzyme-linked immunosorbent assay (ELISA). PMID- 12092835 TI - An enzymatic fluorometric assay for pyridoxal with high specificity and sensitivity. AB - An enzymatic fluorometric assay for pyridoxal with pyridoxal dehydrogenase was developed. The detection limit was about 10 pmol: the calibration curve of pyridoxal showed high linearity (r=0.993). The values obtained by this method correlated well with those by the HPLC method. The enzyme had a high specificity for pyridoxal, and thus animal samples could be directly analyzed without separation of pyridoxal 5'-phosphate by column chromatography. PMID- 12092836 TI - Analysis of the molecular construction of xylogalacturonan isolated from soluble soybean polysaccharides. AB - Soluble soybean polysaccharides (SSPS) extracted from soybean cotyledons are acidic polysaccharides, and exhibited a pectin-like structure. After digesting galacturonan with polygalacturonase, two novel galacturonan (GN) fragments, which were directly linked to xylosyl oligosaccharides, were obtained. One consisted of (beta-D-Xyl)7 branched at the C-3 site of 1,4-linked (alpha-D-GalA)4, and the other consisted of (beta-D-Xyl)4 branched at the C-3 site of 1,4-linked (alpha-D GalA)3. PMID- 12092837 TI - Molecular phylogeny of symbiotic basidiomycetes of fungus-growing termites in Thailand and their relationship with the host. AB - Termitomyces-related symbiotic basidiomycetes in the nests of fungus-growing termites (Macrotermitinae) of several genera in Thailand were cultivated and analyzed phylogenetically based on the DNA sequence of nuclear ribosomal RNA genes. The relationships of the symbiotic fungi with host termites and their locality were apparently complex, supporting intricate mechanisms for the termites to acquire the symbionts. PMID- 12092838 TI - Syntheses of four methyl-branched secondary acetates and a methyl-branched ketone as possible candidates for the female pheromone of the screwworm fly, Cochliomyia hominivorax. AB - 6-Acetoxy-19-methylnonacosane (1), 7-acetoxy-19-methylnonacosane (2), 8-acetoxy 19-methylnonacosane (3), 7-acetoxy-15-methylnonacosane (4), and 21-methyl-7 hentriacontanone (5) were synthesized as racemic and diastereomeric mixtures. These are new compounds isolated from an HPLC fraction of the female hexane extract which elicited mating responses in the male screwworm fly, Cochliomyia hominivorax. PMID- 12092839 TI - Technical improvement to 2D-PAGE of rice organelle membrane proteins. AB - Cytosolic and membrane-associated proteins prepared from rice cells were separated and compared by two different 2D-PAGE methods, isoelectric focusing (IEF)/SDS-PAGE and nonequilibrium pH gradient electrophoresis (NEPHGE)/SDS-PAGE. Although IEF/SDS-PAGE of the cytosolic proteins showed sufficient resolution, some mitochondrial and basic microsomal membrane-associated proteins were weakly or hardly detectable on the 2D gel. High-quality and -quantity separation of the organelle membrane-associated proteins was accomplished by NEPHGE/SDS-PAGE, the advantage of this method being more critical in tightly membrane-bound proteins that were unwashable with NaCl. These results indicate that NEPHGE/SDS-PAGE is a useful tool for the proteomic analysis of rice membrane-associated proteins. PMID- 12092841 TI - The groESL operon of the halophilic lactic acid bacterium Tetragenococcus halophila. AB - The groESL operon of the halophilic lactic acid bacterium Tetragenococcus halophila was cloned by a PCR-based method. The molecular masses of GroES and GroEL proteins were calculated to be 10,153 and 56,893 Da, respectively. The amount of groESL mRNA was increased 3.8-fold by heat shock (45 degrees C), and 4 fold by high NaCl (3-4 M). The Bacillus subtilis sigmaA-like constitutive promoter existed in front of groES, and was used under both normal and stress (heat shock and high salinity) conditions. PMID- 12092840 TI - Method for purification of fluorescence-labeled oligosaccharides by pyridylamination. AB - We developed a convenient method for purification of PA-oligosaccharides to remove contaminants originating from natural fluorescent materials, and excess reagents as well as by-products of tagging reactions in glycan analysis. The method, using a C18-cartridge, is simple and powerful to remove them. Several examples of experiments that showed the usefulness of this purification method are described in this report. PMID- 12092842 TI - Purification and characterization of heparinase that degrades both heparin and heparan sulfate from Bacillus circulans. AB - A heparinase that degrades both heparin and heparan sulfate (HS) was purified to homogeneity from the cell-free extract of Bacillus circulans HpT298. The purified enzyme had a single band on SDS-polyacrylamide gel electrophoresis with an estimated molecular mass of 111,000. The enzyme showed optimal activity at pH 7.5 and 45 degrees C, and its activity was stimulated in the presence of 5 mM CaCl2, BaCl2, or MgCl2. Analysis of substrate specificity and degraded disaccharides demonstrated that the enzyme acts on both heparin and HS, similar to heparinase II from Flavobacterium heparinum. PMID- 12092843 TI - Hypoglycemic effect of a Lentinus edodes exo-polymer produced from a submerged mycelial culture. AB - The hypoglycemic effect of an exo-polymer produced from a submerged mycelial culture of Lentinus edodes was investigated in streptozotocin-induced diabetic rats. The administration of the exo-polymer (200 mg/kg BW) reduced the plasma glucose level by as much as 21.5%, and increased plasma insulin by 22.1% as compared to the control group. It also lowered the plasma total cholesterol and triglyceride levels by 25.1 and 44.5%, respectively. Gel chromatography of the exo-polymer revealed a single peak which is likely to have been a glycoprotein with a molecular weight of 52 kDa and was found to contain 83.5% carbohydrate and 16.5% protein. The Sugar and amino acid compositions of the exo-polymer were analyzed in detail. PMID- 12092844 TI - Biogenesis of 2-phenylethanol in rose flowers: incorporation of [2H8]L phenylalanine into 2-phenylethanol and its beta-D-glucopyranoside during the flower opening of Rosa 'Hoh-Jun' and Rosa damascena Mill. AB - To clarify the biosynthetic pathway to 2-phenylethanol (2), the deuterium-labeled putative precursor, [2H8]L-phenylalanine ([2H8-1]), was fed to the flowers of Rosa 'Hoh-Jun' and R. damascena Mill. throughout maturation, ceasing feeding at the commencement of petal unfurling and at full bloom. Based on GC-MS analyses, [2H8]-1 was incorporated into both 2 and 2-phenylethyl beta-D-glucopyranoside (3) when the flowers were fed until full bloom, whereas no such incorporation into 2 was apparent when feeding was ceased earlier. In both species of rose, the labeling pattern for 2 was almost identical to that for 3, and indicated the presence of [2H6]-, [2H7]- and [2H8]-2, and [2H6]-, [2H7]- and [2H8]-3. This may be ascribed to the equilibrium between 2 and 3. The labeling pattern for 2 and 3 also indicated that these compounds were produced from 1 via several routes, the route involving phenylpyruvic acid being the major one. PMID- 12092845 TI - Identification of a novel anti-ice-nucleating polysaccharide from Bacillus thuringiensis YY529. AB - Strain YY529, capable of producing some anti-ice-nucleating materials (ANM), was isolated from the surface of a camphor leaf. Strain YY529 was identified as Bacillus thuringiensis from its characteristics and taxonomy; the optimum temperature and pH for producing these ANMs were 30 degrees C and 7.0, respectively. One of the ANM with the highest activities among them was purified from the culture. The molecular weight of the ANM was approximately 130 kDa based on a gel filtration analysis. We confirmed that this ANM was a polysaccharide based on the results of the treatment with a mannosidase and the molish reaction. In addition, the LCMS analysis showed that this anti-ice-nucleating polysaccharide (ANPS) had the polyacetyl-D-glucosamine moiety in its structure. Furthermore, this ANPS showed its ability as a non-freeze agent in a preservative solution for the cryopreservation of cock liver. This is the first report of ANPS as a novel ANM from Bacillus thuringiensis YY529. PMID- 12092846 TI - Purification, characterization, and gene sequencing of a catalase from an alkali- and halo-tolerant bacterium, Halomonas sp. SK1. AB - An alkali- and halo-tolerant bacterium with high catalase activity was isolated and identified as a new species of the genus Halomonas. Its catalase (HktA) was simply purified by two steps of liquid chromatography. A 71.4% yield of the catalase was obtained with 97% purity on SDS-PAGE. The specific activity of HktA (57,900 U/mg protein) was two times higher than that of bovine liver catalase. The purified enzyme is inhibited by KCN, NH2OH, NaN3, and 3-amino-1,2,4-triazole, active at pH 5.0-11.0, thermo-sensitive, and KCl-tolerant. HktA is suggested to be a typical catalase, a homotetrameric protein containing heme groups in the active sites. The nucleotide sequence of the catalase gene (hktA) comprises 1,530 bp, encoding a protein of 509 amino acid residues. The deduced amino acid sequence of the hktA shares 99% identity with that of Vibrio rumoiensis S-1T. PMID- 12092847 TI - Antigen presentation by Peyer's patch cells can induce both Th1- and Th2-type responses depending on antigen dosage, but a different cytokine response pattern from that of spleen cells. AB - The Th1 and Th2 preference induced by cells from the Peyer's patch (PP) and spleen (SPL) with various doses of an antigen was examined. The same splenic T cell receptor-transgenic CD4+ T cells were first incubated with PP or SPL cells in the presence of various doses of an antigen, and the cytokine response was observed after secondary stimulation. A Th2-type pattern was only obtained for primary stimulation at 10 microM of the antigen with PP cells, whereas a Th1 pattern was induced at both higher and lower concentrations. SPL cells in the presence of 0.1 to 1 microM of the antigen induced the secretion of Th2-type cytokines. Ten and 100 microM of the antigen plus SPL cells did not induce the release of a large quantity of cytokines. PP cells induced a different cytokine pattern at the antigen concentration that induced a similar level of T cell proliferation with SPL cells. Our findings suggest that the antigen-dose dependent development of Th1/Th2 cells is differentially modulated by the antigen presentation function of cells in PP and SPL. PMID- 12092848 TI - Antifungal activity of rye (Secale cereale) seed chitinases: the different binding manner of class I and class II chitinases to the fungal cell walls. AB - The antifungal activities of rye seed chitinase-a (RSC-a, class I) and -c (RSC-c, class II) were studied in detail using two different bioassays with Trichoderma sp. as well as binding and degradation experiments with the cell walls prepared from its mycelia. RSC-a inhibited more strongly the re-extension of the hyphae, containing mainly mature cells, than RSC-c did. Upon incubation of the fungus with fluorescent chitinases, FITC-labeled RSC-a was found to be located in the hyphal tips, lateral walls, and septa, while FITC-labeled RSC-c was only in the hyphal tip. RSC-a had a greater affinity for the cell walls than RSC-c. RSC-a liberated a larger amount of reducing sugar from the cell walls than RSC-c did. These results inferred that RSC-a first binds to the lateral walls and septa, consisting of the mature cell walls, and degrades mature chitin fiber, while RSC c binds only to the hyphal tip followed by degradation of only nascent chitin. As a result, RSC-a inhibited fungal growth more effectively than RSC-c. Furthermore, it was suggested that the chitin-binding domain in RSC-a assists the antifungal action of RSC-a by binding to the fungal hypha. PMID- 12092849 TI - Purification and characterization of a lipase from the glycolipid-producing yeast Kurtzmanomyces sp. I-11. AB - An extracellular lipase produced by the glycolipid-producing yeast Kurtzmanomyces sp. I-11 was purified by ammonium sulfate precipitation and column chromatographies on DEAE-Sephadex A-25, SP-Sephadex C-50, and Sephadex G-100. Based on the analysis of the purified lipase on sodium dodecyl sulfate polyacrylamide gel electrophoresis, the purified lipase was judged to be homogeneous and its molecular mass was estimated to be approximately 49 kDa. The optimum temperature for the activity was 75 degrees C, and the activity was very stable at temperatures below 70 degrees C. The active pH range of this lipase was 1.9-7.2, and the activity was stable at pH below 7.1. The lipase showed a preference for C18 acyl groups by measurements with p-nitrophenyl esters and triglycerides as substrates. The lipase was very stable in the presence of various organic solvents at a concentration of 40%. Although the N-terminal sequence of the Kurtzmanomyces lipase was very similar to that of lipase A from Candida antarctica, the pH profiles of the two lipases were significantly different. PMID- 12092851 TI - Isolation and culture conditions of a Klebsiella pneumoniae strain that can utilize ammonium and nitrate ions simultaneously with controlled iron and molybdate ion concentrations. AB - A bacterial strain F-5-2, isolated from soil and identified as Klebsiella pneumoniae, removed NH4+ completely in 24 h of aerobic cultivation in a medium containing 1 mg/ml of NH4NO3. However, 70% of the NO3- originally provided remained. When 100 microM Fe2+ was added to the medium, both NH4+ and NO3- were removed simultaneously and completely from the culture within 6 h of incubation. In addition, the amount of MoO4- in the medium markedly affected the bacterial cell growth and utilization of NH4+ and NO3-. The bacterium could remove 4 mg/ml of NH4NO3 completely in 48 h of aerobic cultivation in a medium containing 100 microM Fe2+ and 0.8 pM MoO4(2-). The total nitrogen in the culture containing its cells was decreased to 14% of that in the NH4NO3 originally provided. GC-MS analysis demonstrated that N2 was generated from the nitrogen atoms of both NH4+ and NO3-. PMID- 12092850 TI - Gene cloning and biochemical analysis of thermostable chitosanase (TCH-2) from Bacillus coagulans CK108. AB - The DNA sequence of the thermostable chitosanase TCH-2 gene from Bacillus coagulans CK108 showed a 843-bp open reading frame that encodes a protein of 280 amino acids with a signal peptide corresponding to 32 kDa in size. The deduced amino acid sequence of the chitosanase from Bacillus coagulans CK108 has 61.6%, 48.0%, and 12.6% identities to those from Bacillus ehemensis, Bacillus circulans, and Bacillus subtilis, respectively. C-Terminal homology analysis shows that the enzyme belongs to the Cluster I group. The size of the gene was similar to those from mesophiles of the Cluster I group with regard to higher preference for codons ending in G or C. The recombinant chitosanase was electrophoretically purified to homogeneity by only two steps with column chromatography. The half life of the enzyme was 40 min at 90 degrees C. The purified protein was also highly stable, retaining above 50% residual activities during treatment with denaturants such as urea (8 M) and guanidine x HCl (4 M) at 37 degrees C for 30 min. The enzyme had a useful reactivity and a high specific activity for producing functional oligosaccharides as well, producing the tetramer as a major product. PMID- 12092852 TI - Evaluation of a preparatory community-based prostate health education program. AB - BACKGROUND: Medical organizations are divided on the issue of screening for prostate cancer, yet there is agreement that men should be educated about prostate health. Shared decision making involves patients and practitioners in this process, yet some men need preparatory education prior to the physician encounter. This study assessed the effectiveness of a community prostate health awareness program, focusing on men at risk for prostate cancer. METHODS: Participants were given a pretest and a posttest to assess knowledge gains and impact on short-term intentions to address their prostate health. RESULTS: There was a statistically significant increase in knowledge. Short-term intentions increased for those participants meeting the inclusion criteria. DISCUSSION: Community outreach programs remain an excellent vehicle to educate the public and complement the efforts of health care providers. PMID- 12092853 TI - Reflective...practice. PMID- 12092854 TI - Palliative Care Section. PMID- 12092855 TI - Development of a cancer genetics education program for clinicians. AB - BACKGROUND: There is a gap in knowledge about hereditary cancer and genetic testing among primary care practitioners. Education is needed to enable identification and management of patients at high risk for cancer. METHODS: A new cancer genetics curriculum was delivered through individual lectures and full-day conferences. Innovative marketing and conference organizational approaches were used to increase participation. RESULTS: The curriculum has been delivered to 7,400 health care professionals with diverse educational backgrounds. CONCLUSION: Conventional formats were successfully used to implement this new curriculum. CME evaluations indicated satisfaction with the programs and a clear need for and continued interest in cancer genetics applications. PMID- 12092856 TI - Breast cancer genetics education for college women: an evaluation of approaches. AB - BACKGROUND: A strong family history of breast cancer may greatly increase the chances for young women to develop the disease. Breast cancer genetics education for college-age women would increase awareness, but an effective method remains elusive. METHODS: Thirty-five college women viewed four education tools (CD-ROM, video, brochure, and lecture) and discussed topic importance, tool preference, and distribution ideas in focus groups. RESULTS: The participants considered breast cancer genetics important; however, the information must be made relevant to their peers to be accepted. CONCLUSION: An ideal education method would focus on general breast health, mentioning family history as an important risk factor. PMID- 12092857 TI - A concept model for individual surveillance in genetic cancer care. AB - OBJECTIVE: To present the concept of individual surveillance in genetic risk management as a complex collection of other concepts, and to propose a model that represents how these component concepts may relate to each other. CONCEPT: The concept of "individual surveillance" has a set of lexical conditions, called critical attributes, antecedents, and consequence. The attributes are derived from the historical and contemporary use of the word and must be present for conceptualization. Antecedents are conditions that must exist before individual surveillance can occur, and consequences are those events that will always occur after individual surveillance. The popular association of surveillance with real or suspected criminal activity may design a predilection for critical attributes and confuse concept attainment. Before involving patients and family members in surveillance activities, clinicians should explore the subjective meaning of participation. MODEL: The model can be used, by a clinician and a patient, to describe and understand the meaning of individual surveillance for participants in a systematic, holistic, and highly personal way. PMID- 12092858 TI - Graduating dental students' perceptions of oral cancer education: results of an exit survey of seven dental schools. AB - METHOD: A 22-question exit survey was administered on a voluntary basis to senior dental students at seven dental schools in order to evaluate perceptions regarding their knowledge and skills in oral cancer control. The questions encompassed prevention, role of tobacco, lesion recognition, diagnostic techniques, and patient management. RESULTS: The range of responses was broad between students as well as schools regarding didactic and clinical exposures, and confidence in abilities to manage oral cancer patients. CONCLUSION: Data from this survey showed a perceived lack of knowledge and skills among graduating dental students that may translate to a subsequent deficiency in incorporating optimal oral cancer control procedures in their practices. PMID- 12092859 TI - An interdisciplinary cancer prevention and control training program in public health. AB - Description. From 1988 through September 2001 the Cancer Prevention and Control Training Program at the University of Alabama at Birmingham has supported 52 pre- and postdoctoral trainees in Public Health disciplines, with 31% of the trainees members of minority groups (21% African Americans). Strengths of the program are director, co-director, and advisor accessibility, latitude in selecting research topics, and focus on prevention and control. Public health trainees have co authored 135 publications and made 141 other professional contributions. The program plans to add trainees in outcomes research and a second advisor for each trainee to enhance its inherent interdisciplinarity. PMID- 12092860 TI - Technology available in nursing programs: implications for developing virtual end of-life educational tools. AB - BACKGROUND: To overcome insufficient attention to end-of-life (EOL) care in nursing education, the authors are developing the "Toolkit for Nursing Excellence at End-of-Life Transition" (TNEEL). METHOD: An evidence-based design process was used to create a computerized (CD-ROM) multimedia toolkit of instructional aides. An online survey of all U.S. undergraduate nursing programs was conducted to identify their current technologic infrastructures available for innovative teaching aides. Expert review process guided pedagogic decisions. RESULTS: Survey findings enabled TNEEL development to be responsive to nurse educators' needs and preferences. CONCLUSION: Educators can use TNEEL to actively engage students in meaningful learning about EOL care. PMID- 12092861 TI - Beyond doughnuts: tumor board recommendations influence patient care. AB - BACKGROUND: There is little solid evidence to support the belief that tumor conferences directly impact the care patients receive. This study investigated the recommendations made at tumor conferences and whether these recommendations were enacted. METHODS: Cancer registrars from Oregon-area hospitals with tumor boards that utilized a prospective "working" format were surveyed regarding tumor board characteristics and content. They also recorded the recommendations made for each prospective case presented at each tumor board for one month. Later, they contacted each patient's provider to see which recommendations had been implemented. RESULTS: Information from eight tumor boards in six hospitals was recorded. The institutions varied with regard to size, location, number of tumor boards, number of cases reviewed annually, and types of cancers reviewed. One hundred fifty-three specific, prospective recommendations were made for the care of 97 patients. Of these, 128 (84%) were followed, 2 (1%) were pending, and 5 (3%) were status unknown. Of recommendations followed, 21 (16%) were for diagnosis, 100 (78%) were for therapy, and 7 (5%) were for palliation. CONCLUSION: Recommendations made at tumor conferences are generally implemented into patient care. Tumor boards can play a strategic role in the planning of care for cancer patients. PMID- 12092862 TI - The history and evolution of surgical instruments: XI retractors, dilators and related inset pivoting instruments. PMID- 12092863 TI - Peri-operative fluid and electrolyte management: a survey of consultant surgeons in the UK. AB - BACKGROUND: Current peri-operative fluid and electrolyte management in the UK may be suboptimal. We assessed the attitudes of consultant surgeons to fluid and electrolyte prescribing and gathered suggestions for improvement in education on the subject. METHODS: A postal questionnaire survey was sent to 1091 Fellows of the Association of Surgeons of Great Britain and Ireland. Of the 730 (67%) replies, 20 were invalid or incomplete, and 710 (65%) questionnaires were analysed. Outcome measures included provision of guidelines and teaching to junior staff on fluid and electrolyte prescribing, appropriateness of fluid management and suggestions to improve standards. RESULTS: Junior staff were given written guidelines in 22% of instances. Only 16% of respondents felt that their preregistration house officers (PRHOs) were adequately trained in the subject before joining the firm; 15% also stated that PRHOs did not receive much training on their firm. 65% felt that fluid balance charts were accurately maintained, nursing shortages being the commonest reason for inaccuracies. Only 30% felt that postoperative patients were receiving appropriate amounts of water, sodium and potassium. Respondents who had been consultants for > 5 years were more likely to prefer erring on the side of under-replacement of fluid than those who were consultants for 5 years (63% versus 47%, P < 0.0005). Suggestions for improvement in education included problem-oriented ward rounds, written guidelines, and discussion of patient scenarios. CONCLUSIONS: Consultant surgeons feel that present practice in peri-operative fluid management is unsatisfactory. Higher standards within clinical governance and risk management may be achieved by focused practical training combined with formal written guidelines. PMID- 12092864 TI - Video-assisted thoracic surgery for mediastinal extramedullary haematopoiesis. AB - Extramedullary haematopoiesis is a rare cause of an intrathoracic mass. We report a case of posterior mediastinal extramedullary haematopoietic mass in a 50-year old man who presented with non-specific symptoms and a paravertebral mass on chest X-ray. Diagnosis was achieved by using video-assisted thoracic surgery. PMID- 12092865 TI - Conservative management of tension pneumoperitoneum. AB - Tension pneumoperitoneum is characterized by a rapidly increasing intra-abdominal pressure, the haemodynamic and respiratory consequences of which are potentially fatal. Prompt laparotomy, surgical drainage and identification of possible sites of perforation is usually advocated in spite of previous reports of negative laparotomies in the literature. We report the successful non-surgical management of two cases of tension pneumoperitoneum. PMID- 12092866 TI - Surgery for pancreas divisum. AB - We present our experience of open surgical treatment in 5 patients with symptomatic pancreas divisum (PD). Choice of therapy was based on allocation of patients to one of five clinical presentation groups: (i) with minor symptoms (no operation); (ii) with recurrent acute pancreatitis or upper abdominal pain (RAP/RUAP)--3 patients; (iii) with radiological evidence of chronic pancreatitis (CP)--1 patient; (iv) chronic pancreatic pain without radiological evidence of chronic pancreatitis (CPP); and (v) other pancreatic complications--1 patient. This classification helps to decide management and predict possible outcome. Various types of operation were performed as indicated (open surgical accessory sphincteroplasty [2 also had distal pancreatectomy], n = 3; Puestow's operation, n = 1; or Beger's pancreatectomy, n = 1). All patients improved significantly and are now leading normal personal, professional, and social lives. We conclude that, with careful selection of patients and appropriate therapy, the response to surgical treatment is good. PMID- 12092868 TI - Rubber band ligation of haemorrhoids in the out-patient clinic. AB - Rubber band ligation (RBL) is an effective treatment for symptomatic haemorrhoids but carries significant morbidity. We performed a prospective study of 98 consecutive patients treated by RBL in the out-patient clinic. Immediate, intermediate (within 2 weeks) and late (within 2 months) complications were recorded. Immediate complications occurred in 66 (67.3%) patients. Pain was the predominant symptom in 50 patients (51%). Fifteen (15.3%) patients had vasovagal attacks and 1 (1%) had bleeding. Twenty-five patients (25.5%) were unable to perform normal activities on the day of RBL. One patient needed hospital admission for control of pain. Seventy four (75.5%) patients would have RBL if they needed further treatment for haemorrhoids. Symptomatic cure was achieved in 71 patients (72.4%). RBL is an effective treatment but with significant complications. Patients should be adequately warned, especially of pain and vasovagal attacks. PMID- 12092867 TI - Compartment syndrome following pelvic surgery in the lithotomy position. AB - We report the cases of two patients who developed compartment syndrome following pelvic surgery in the lithotomy position. These cases highlight this important and potentially devastating complication. PMID- 12092869 TI - Complete small bowel recovery after prolonged total venous occlusion. AB - Acute small bowel ischaemia is a mostly irreversible condition associated with high mortality. Here we report the case of a patient after severe abdominal trauma in whom the superior mesenteric vein (SMV) was completely occluded for more than 15 h in the absence of any collateral venous drainage. Following surgical reconstruction of the SMV and with scheduled relaparotomies for 5 days, the bowel showed slow recovery. Now the patient is well and on complete oral nutrition. PMID- 12092870 TI - Spontaneous resolution of a superior mesenteric vein thrombosis after laparoscopic nissen fundoplication. AB - A previously fit 37-year-old man developed superior mesenteric venous thrombosis after undergoing a laparoscopic Nissen fundoplication. Despite receiving thrombo embolic prophylaxis on postoperative day 16, he presented with a gradual onset of vague, but severe, umbilical and epigastric pains. Laboratory tests, abdominal ultrasound scan and gastroscopy were all unremarkable. Contrast enhanced abdominal spiral computerised tomography (CT) revealed a partial occlusion of the superior mesenteric and portal vein due to a thrombus; abnormal flow was confirmed on colour Doppler ultrasound. A predisposing hyper-coagulable condition was excluded. The patient responded rapidly to expectant management and a repeat spiral CT scan, 3 weeks after the initial presentation, demonstrated complete re canalisation of the vessel. Although rare, superior mesenteric venous thrombosis is probably underdiagnosed due to the vague nature of the symptoms, the lack of clinical signs, a low index of suspicion on the part of the clinician, and then subsequent failure to request the optimal investigation--namely contrast enhanced abdominal spiral CT scan. We discuss the possible mechanisms by which laparoscopic surgery may increase the risks of developing superior mesenteric venous thrombosis, the pitfalls in diagnosis and treatment options. PMID- 12092871 TI - Medicolegal claims in vascular surgery. AB - BACKGROUND: Knowledge about medicolegal claims is important for risk management and clinical practice. This study presents the first comprehensive collated data for vascular surgical practice. METHOD: Details of claims notified to the National Health Service Litigation Authority (NHSLA) since its inception in 1995, and to the Medical Defence Union (MDU) from 1990-1999 were analysed. RESULTS: A total of 424 claims were notified--170 NHSLA and 254 MDU (176 from surgeons who described themselves as 'vascular' and 248 from 'general surgeons'). Varicose veins were the commonest condition involved (244 claims) and nerve damage was the most frequent complaint (76), followed by incorrect or unsatisfactory surgery (35), and damage to the femoral vein (16) or artery (13). Arterial claims (174) against vascular surgeons comprised 88% in the NHS but only 39% in private practice: 45 related to aortic grafting, 28 to other bypass grafts, and 36 alleged failure to recognise or treat ischaemia. CONCLUSIONS: The likely cause of many of these claims was failure to advise patients about potential risks and expected benefits. Recognition of the areas of highest risk, with improvements in communication and record keeping, may limit future claims. PMID- 12092872 TI - Audit of attendance rates for an infra-inguinal graft surveillance programme--how complete is your graft surveillance? AB - Much has been written on the subject of graft surveillance following infra inguinal bypass graft surgery, but surprisingly little information exists on actual attendance rates for such programmes. The aim of this study was to calculate the attendance rate for graft surveillance at the vascular unit in Cheltenham. All patients who entered the graft surveillance programme following an infra-inguinal graft procedure over a 5-year period were included in the study. Partial and complete surveillance attendance rates were calculated for each year in the study period. Both rates were seen to rise over the 5-year period, from 31% to 78% and 0 to 38%, respectively. These attendance rates were much lower than were expected. Vascular units relying on such graft surveillance programmes should evaluate their own 'surveillance rates'. PMID- 12092873 TI - The internet--friend or foe? A questionnaire study of orthopaedic out-patients. AB - OBJECTIVE: To examine patients' use of the Internet to obtain medical information, their opinions on the quality of medical Web sites, and their attitudes towards Internet-based consultations. DESIGN: Questionnaire study. PARTICIPANTS AND SETTING: 398 patients, aged 10-95 years, visiting the orthopaedic outpatient clinics of a London district general hospital over a 2 week period. MAIN OUTCOME MEASURES: (i) The rate of Internet use by patients; (ii) the perception of the quality of medical web sites; (iii) future intentions and attitudes towards Internet-based consultations; and (iv) concurrence between information obtained from Web sites and advice given by the orthopaedic surgeon in the clinic. RESULTS: From 369 respondents (response rate 93%), 55.3% of patients had accessed the Internet. Of these, 52.0% had obtained medical information from this source. Access was linearly correlated with age (r2 = 0.975, P < 0.01) and was also related to social status. Of the 12.3% of patients who had researched their particular orthopaedic condition, 20% reported that the advice received from the surgeon in the clinic contradicted that obtained from the Internet. A total of 35.7% of patients would undergo an Internet-based consultation, whilst a further 25.5% would consider this, depending on the medical condition in question. CONCLUSIONS: Over half of the patients studied were willing to access the Internet for medical information, with younger patients more likely to undertake this activity. Moreover, a significant proportion of respondents were willing to undergo an Internet-based consultation. The increased use of medical Web sites by patients raises important issues regarding the need for quality control, and impacts significantly upon the surgeon-patient relationship. PMID- 12092874 TI - The use and availability of thyroid shields in orthopaedic theatres: a telephone questionnaire of English hospitals. AB - There are no published guidelines issued for the use of thyroid shields by either the Royal College of Radiologists or the British Orthopaedic Association. It has previously been demonstrated that a thyroid shield should be worn during fluoroscopic screening using a portable image intensifier. Scrub staff not wearing a thyroid shield are currently being exposed to ionising radiation on a regular basis, with potentially harmful effects. Of the 210 hospitals in England with orthopaedic departments, 179 were telephoned and the use and availability of thyroid shields was asked. The results demonstrated that 98 orthopaedic theatres had sufficient numbers of thyroid shields available. Of these 98 orthopaedic theatres, thyroid shields were routinely used in only 28 theatres during fluoroscopic screening on a regular basis. It is the authors' recommendation that thyroid shields should be worn during orthopaedic procedures involving the use of an image intensifier. PMID- 12092875 TI - A 5-year study of the outcome of surgically treated depressed skull fractures. AB - BACKGROUND: Many changes and improvement have taken place in the management of head injured patients in the last 20 years. There have been few recent studies analysing the overall outcomes including early complications of depressed skull fractures. The aim of our study was, therefore, to examine the factors influencing the surgical outcome of patients with depressed skull fractures. METHODS: We reviewed case notes of 73 consecutive surgically treated depressed skull fractures during the period from 1 January 1994 to 31 December 1998 admitted to the Walton Centre for Neurology and Neurosurgery, Liverpool. RESULTS: There was a male preponderance of 9:1. Alleged assault was the most common cause of depressed skull fractures followed by road traffic accidents. Postoperative infection rate was 8.2%. More than 80% of patients received prophylactic antibiotics. We failed to show any statistically significant association between the use of antibiotics and reduction of the rate of infection. However, prevalence of infection was significantly associated with brain contusion, low GCS score and dural tear (P < 0.05). Prevalence of early post-traumatic epilepsy was 12.3%. No patients received prophylactic anticonvulsants. There was no significant association between dural tear and prevalence of post-traumatic epilepsy. Mortality rate was 1.4%. CONCLUSIONS: Paediatric populations have better outcome; 7 out of 10 patients in this series progressed to full recovery. Use of prophylactic antibiotics did not reduce the infection rate. Presence of dural tear was not associated with an increase risk of post-traumatic epilepsy. PMID- 12092876 TI - Through the letter box--a traumatic hand injury. PMID- 12092877 TI - Urinary tract ultrasonography in the evaluation of haematuria--a report of over 1,000 cases. AB - Over a 5-year period, 1007 patients with haematuria were investigated, using a protocol based on ultrasonography as the upper tract imaging modality of choice. Intravenous urography (IVU) was only used in selected individuals, including those patients with bladder cancer suspected on cystoscopy, suspicious or malignant cytology, previous investigation for haematuria, on-going haematuria at the time of their clinic visit, a history of flank pain or hydronephrosis on ultrasonography. Of this series, 840 (83%) had visible haematuria, 158 (15%) had microscopic or chemical haematuria and 9 (0.9%) had unspecified haematuria. A total of 133 bladder transitional cell tumours, 21 renal cell cancers and 2 upper tract transitional cell cancers (TCC) were diagnosed. The sensitivity of ultrasound with respect to bladder cancer was 63% and the specificity 99%. The odds ratio of diagnosing cancer in patients with visible haematuria compared to microscopic or unspecified haematuria was 3.3. No upper tract tumours were missed using this investigational protocol. An ultrasonography-based protocol could miss fewer upper tract TCCs than a standard IVU-based service would miss renal cell cancer. Provided there is no history of flank pain, no malignant cytology, no hydronephrosis and no previously investigated haematuria, IVU could be safely omitted. PMID- 12092878 TI - Gelofusine allergy--the need for identification jewellery. AB - A case of anaphylactoid reaction due solely to the use of Gelofusine in a patient with non-haemorrhagic hypovolaemia is presented, with a discussion on the management and the use of allergy identification jewellery. PMID- 12092879 TI - Examination of the shoulder. PMID- 12092880 TI - Improvement in surgical exposure of the hip using the direct lateral approach. PMID- 12092881 TI - Clipping perforators without dividing them could reduce postoperative pain and swelling following subfascial endoscopic perforator surgery. PMID- 12092882 TI - A simple technique of irrigating the abdomen and pelvis through an appendicectomy wound. PMID- 12092883 TI - Is transcranial Doppler of use in carotid surgery? PMID- 12092884 TI - Open access carotid duplex scanning. PMID- 12092885 TI - Topical adhesive as a wound dressing for elective abdominal surgery. PMID- 12092886 TI - Promoting quality school physical education: exploring the root of the problem. PMID- 12092888 TI - Hurdle preflight in springboard diving: a case of diminishing returns. AB - Attempts to increase dive height by introducing a period of flight in the final approach step preceding the hurdle of dives from forward and reverse groups were investigated. One study involved 11 collegiate-level divers experienced in both traditional and hurdle preflight techniques. In a second study, dives executed by 9 national-level women were compared. Dives with a hurdle preflight had shorter final approach steps and greater hurdle flight durations. Flight time differences in favor of hurdle preflight techniques diminished from final approach step through hurdle flight to dive flight. Although the collegiate-level divers had longer dive flight times when using a hurdle preflight, it was suggested that the costs of these techniques may outweigh their potential benefits. PMID- 12092887 TI - The influence of midsole cushioning on mechanical and hematological responses during a prolonged downhill run. AB - The purpose of this study was to investigate the influence of midsole durometer on mechanical and hematological responses during a prolonged downhill run. Twenty four men completed a 30-min downhill run (-12% grade) wearing either soft, medium, or hard midsole shoes. Data describing mean peak tibial acceleration (PTA), stride frequency, plasma free hemoglobin, hemoglobin concentration, hematocrit, and creatine kinase (CK) were collected. While there were no significant differences in PTA among midsole durometer shoes, PTA increased by 20% after the first 5 min of the run over all other time intervals during the run (p < .05). Hemolysis showed a 50.2% increase from prerun to postrun values (p <.05). CK increased from the prerun state to 24 hr after the run (p <. 05). Downhill running, irrespective of midsole durometer, showed increased levels of legshock, hemolysis, and muscle damage over values that are present in the literature for a level running protocol. PMID- 12092889 TI - Perceptions of the physical environment, stage of change for physical activity, and walking among Australian adults. AB - Data for this study were from a population survey of 1,200 adults ages 40-60 years, sampled from the Illawarra region of New South Wales. Questionnaire items on perceptions of the environment were factor analyzed into: Aesthetic Environment and Practical Environment. For both factors, those in the Contemplation stage had more negative views of the environment than those in Maintenance. Those who walked for 0-20 min/week held more negative perceptions of their environment than those who walked for 21-120 min/week and those who walked for > 120 min/week. The health promotion implications of these findings are that environments perceived as attractive and as providing convenient access to services and facilities may influence motivational readiness for physical activity and time spent walking. PMID- 12092890 TI - Visual orienting in college athletes: explorations of athlete type and gender. AB - Covert orienting was measured in 50 college athletes and 51 nonathletes of both genders. Visual environments of the sports were both static (swimming, track) and dynamic (soccer, volleyball). Participants made speeded responses in a task measuring vigilance, alerting, automatic orienting, voluntary orienting, modulation of automatic orienting, and modulation of inhibition of return. Gender differences werefound in the overall response times of nonathletes and in the alerting measures for all participants. However, all participants were similar in their automatic orienting. Sport-specific effects were seen in voluntary orienting and in the modulation of automatic orienting. These gender and sports related findings are interpreted in light of the experience athletes have in the dynamic control of spatial attention. PMID- 12092891 TI - Application of the LaGrange polynomial in skeletal muscle fatigue analysis. AB - The percentage of decrement in torque and the number of serial contractions are mutually exclusive methodological controls in the study of muscular fatigue. This paper examines the feasibility of using the LaGrange polynomial in the analysis of voluntary muscular fatigue patterns. Twenty-one men (ages 20-60 years) reported to the orthopedic biomechanics laboratory on 2 days separated by 4 months. During both sessions, participants completed three maximal isokinetic (180 deg x s(-1)) contractions of the knee extensors to serve as baseline, before starting the fatigue protocol. The fatigue protocol consisted of serial contractions until a 50% strength decrement was reached. The LaGrange polynomial was first used to interpolate the individual fatigue pattern for each participant into 15 data points (trials). Data analysis was then conducted on these 15 data points. Intraclass correlation analysis of variance showed that the reliability of baseline torque was very good (.93). Baseline torque, the average of three trials, exhibited a 5.4 Nm (6%) increase from the first to second test session (p < .05). The mean level of torque, average of the 15-point fatigue pattern, also increased 7.5 Nm (15%) on the second test session (p < .05). The classic torque deficit for the first trial of a fatigue series was preserved by the interpolation method. Serial contractions resulted in an average decrease in torque of 29.5 Nm (50%) from the first to last trial (p <.05). The interpolation method also retained the linear and quadratic trend components commonly observed for isometric and isokinetic fatigue patterns. The two trend components accounted for 94.7% of the total trial variance. It was concluded that the LaGrange polynomial used to interpolate fatigue patterns to fewer data points was successful. PMID- 12092892 TI - A survey of U.S. Atlanta and Nagano Olympians: variables perceived to influence performance. AB - The purpose of this study was to determine the frequency and magnitude of specific variables perceived to have affected U.S. Olympic athlete performance. Participants included 296 Atlanta Olympians and 83 Nagano Olympians. Olympians rated how they perceived specific variables influenced their Olympic performance. Results revealed that numerous variables, including performance influences, such as preparation for distractions and loss of composure; team variables, such as strong cohesion and positive coach-athlete relationships; coaching variables, including coach's ability to deal with crises and coaching expectations; family friend variables, including general social support and getting event tickets for family and friends; and environmental concerns, such as venue transportation difficulties and Olympic village distractions; were perceived to influence performance. Findings verified the results of qualitative interviews conducted with Olympic athletes and coaches. PMID- 12092893 TI - Participation in extracurricular physical activity programs at middle schools. PMID- 12092894 TI - Extending the theory of planned behavior in the exercise domain: a comparison of social support and subjective norm. PMID- 12092895 TI - Physical activity patterns in recent college alumni. AB - Within this sample of recent college alumni, a high proportion indicated that they enjoy exercise (66.1%) and felt confident in setting their own fitness programs (79.4%). Yet, only 34.3% reported engaging in recommended levels of moderate or vigorous PA. In contrast to previous reports, rates of participation in vigorous, moderate, and strengthening-type PA did not differ between men and women. Alumni who reported that they engaged in adequate levels of moderate or vigorous PA had gained significantly less weight in the years following graduation than those who reported being insufficiently active (3.0 kg vs. 6.1 kg). Last, we noted a strong positive association between PA patterns as college seniors and current PA behavior as recent alumni. PMID- 12092896 TI - Association of parent and peer support with adolescent physical activity. PMID- 12092897 TI - Evaluating the factorial structure of the revised causal dimension scale in adolescents. PMID- 12092898 TI - A comparison of video and text-based questionnaire methods for testing older adults. PMID- 12092899 TI - Perceived leg extension and flexion forces of young adult men and women: comparison to previous findings. PMID- 12092900 TI - Trap a gene and find out its function: toward functional genomics in Drosophila. AB - Many declared aims of the genome projects have been achieved. The total genomic sequences of several relatively noncomplex/complex organisms (such as E. coli, yeast, Caenorhabditis, Drosophila) are being determined, and the nucleotide sequencing of the entire human genome will be complete in the near future. However, this achievement is not the end of the road but rather the first step toward the functional understanding of the genome of humans and other organisms. The determined linear nucleotide sequences remain only lists of A, C, G and T, unless they are given functional significance. The coding sequences of genes can be identified in a relatively reliable manner by computational methods, but the exact function of their protein products can rarely be determined without obtaining much additional information, e.g., by biochemical or cell biological methods. Thus, following sequencing, the next step must be to assign functions to the identified genes. The final goal of genome research today may look futuristic, but the knowledge of the function of every single gene and the interactions between them will finally allow us to understand the development and functioning of an organism as a whole. Gene-trapping methodology is a powerful strategy for cloning and identifying functional genes, as it marks a gene with a tag and simultaneously generates a corresponding genetic variation for that particular locus. Therefore, gene trapping is an extremely useful tool for functional genomics, establishing a correlation between the physical and genetic maps of the genome. The relative simplicity of its genome and the availability of huge bodies of genetic and molecular information make Drosophila melanogaster one of the most important model organisms. Its genome will serve as a "reference" for the in-depth analysis of the organization of more complex eukaryotic genomes. Multifaceted approaches to Drosophila functional genomics and the dual-tagging gene trap system newly developed for functional analysis of Drosophila genes are discussed in this review. PMID- 12092901 TI - The gene search system: its application to functional genomics in Drosophila melanogaster. AB - In Drosophila melanogaster, gain-of-function mutagenesis utilizing the GAL4-UAS system has been established, allowing identification of genes that may not be easily detectable by loss-of-function screening approaches. The conditional features of misexpression systems are especially useful for studying late-stage biological processes, such as those involving adult behavior or lifespan. The gene search system, incorporating a bidirectional misexpression vector, was used to screen for genes critical for longevity determination. We have identified several genes whose misexpression in adulthood extends the fly's lifespan. Phenotypic characterization of fly lines carrying a mis-expression vector, in conjunction with obtaining information about the genomic insertion sites, creates valuable resources for the systematic functional genomics in Drosophila. PMID- 12092902 TI - Transposable element-mediated transgenesis in insects beyond Drosophila. AB - In the last few years, cases of transformation involving insects other than Dipterans have been reported. Although transgenics have been created only in a few species, transposable element vectors may be successfully developed in most insect forms in the near future. The major remaining problems revolving round transformation in wide-ranging species of insects are mainly related to methods of DNA delivery. Transposable element-mediated gene transfer in non-Drosophila insects is reviewed. In addition, the current status of honeybee transformation will be explained as an example of an insect transgenic system that faces substantial obstacles to the creation of germ-line transformants. PMID- 12092903 TI - Rnai: a new technology in the post-genomic sequencing era. AB - The complete genome sequences and huge numbers of predicted gene sequences from many complex organisms are available. Reverse-genetic analyses of these organisms will now be necessary to understand what all these genes are doing and how their functions interact. RNAi technology is very useful in this regard for studying gene function, not only in these model organisms, but also in the organisms previously considered not to be amenable to genetic analysis. This treatment reviews the discovery of RNAi, advances in the study of the mechanisms by which this material impinges on gene-product expressions, and technical improvements that will allow RNAi to be applied to a wide variety of organisms. PMID- 12092904 TI - The power law distribution for walking-time intervals correlates with the ellipsoid-body in Drosophila. AB - The temporal properties of a variety of behavioral traits obey power law distributions, a property often referred to as fractal. We recently showed that the temporal pattern of locomotor activity of the fruitfly Drosophila melanogaster follows this distribution. Although an increasing number of such fractal patterns are being discovered, the brain areas and neuronal networks responsible remain unknown. In this study, we show that specifically blocking synapses established by neurons of the Drosophila ellipsoid-body, a substructure of the central complex in the brain, leads to a loss of the fractal properties in the temporal pattern. We conclude that the temporal fractal pattern of locomotor activity is regulated in the ellipsoid-body. PMID- 12092905 TI - Giant neuron pathway neurophysiological activity in per(0) mutants of Drosophila melanogaster. AB - In Drosophila melanogaster, the clock gene period (per) has a clearly defined role in the molecular machinery involved in generating free-running circadian rhythms. per mutations also influence rhythms in the Drosophila love song and in the ultradian timescale. The relationship between these two phenomena has so far escaped satisfactory explanation. Here we analyzed the neurophysiological activity of the giant fiber neural pathway in per(0) flies. Under constant light, and at relatively low stimulation frequencies (1-2 Hz), per(01) flies habituate significantly earlier than they do under 12 h light-dark cycles. The results suggest an involvement of per in phenomena of short-term neural plasticity. PMID- 12092906 TI - Penetrance and expressivity of genes involved in the development of epilepsy in the genetically epilepsy-prone rat (GEPR). AB - In order to understand the level of complexity of the epileptic phenotype in the two strains of Genetically Epilepsy-Prone Rats (GEPRs), we determined two important measures of genetic complexity, penetrance and expressivity. Penetrance is the percentage of animals of a specific genotype who express the phenotype associated with that underlying genotype. Expressivity refers to the degree that a particular genotype is expressed as a phenotype within an individual. Incomplete penetrance and variable expressivity are caused by genetic and environmental variation. In this paper we have studied the epileptic phenotype for 20,373 rats. Animals were tested on three occasions for audiogenic seizure and given an audiogenic response score (on a scale of 0-9, 0 being no seizure and 9 being the most severe). The GEPR-3 and GEPR-9 animals both show incomplete penetrance and variable expressivity of the underlying genetic predisposition. The GEPR-9 strain has more animals that have variable levels of seizure predisposition (as measured by a scoring system that denotes the severity of generalized tonic/clonic seizures) and a greater percentage of animals that exhibit no susceptibility to such seizures induced by sound. Both strains have a number of animals that are not susceptible to sound-induced GTCSs and that exhibit some variability in seizure severity. The GEPR-9 males show greater differences in expressivity and penetrance compared to GEPR-9 females. The GEPR-3 animals also show sex-associated variable penetrance and expressivity of the epileptic phenotype, although the differences are much smaller. These findings are the first step toward the mapping of the underlying quantitative trait loci (QTL) for seizure in these animals. PMID- 12092907 TI - Association analysis of chromosome 15 gabaa receptor subunit genes in autistic disorder. AB - Gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the brain, acting via the GABAA receptors. The GABAA receptors are comprised of several different homologous subunits, forming a group of receptors that are both structurally and functionally diverse. Three of the GABAA receptor subunit genes (GABRB3, GABRA5 and GABRG3) form a cluster on chromosome 15q11-q13, in a region that has been genetically associated with autistic disorder (AutD). Based on these data, we examined 16 single nucleotide polymorphisms (SNPs) located within GABRB3, GABRA5 and GABRG3 for linkage disequilibrium (LD) in 226 AutD families (AutD patients and parents). Genotyping was performed using either OLA (oligonucleotide ligation assay), or SSCP (single strand conformation polymorphism) followed by DNA sequencing. We tested for LD using the Pedigree Disequilibrium Test (PDT). PDT results gave significant evidence that AutD is associated with two SNPs located within the GABRG3 gene (exon5_539T/C, p=0.02 and intron5_687T/C, p=0.03), suggesting that the GABRG3 gene or a gene nearby contributes to genetic risk in AutD. PMID- 12092908 TI - Coping, functioning, and adjustment of rescue workers after the Oklahoma City bombing. AB - Studies have not previously considered postdisaster adjustment in the context of psychiatric disorders. After the Oklahoma City bombing, a volunteer sample of 181 firefighters who served as rescue and recovery workers was assessed with a structured diagnostic interview. The firefighters had relatively low rates of posttraumatic stress disorder (PTSD) and described little functional impairment, positive social adjustment, and high job satisfaction. PTSD was associated with reduced job satisfaction and functional impairment, providing diagnostic validity. Turning to social supports, seeking mental health treatment, and taking medication were not widely prevalent coping responses. Postdisaster alcohol use disorders and drinking to cope were significantly associated with indicators of poorer functioning. Surveillance for problem drinking after disaster exposure may identify useful directions for intervention. PMID- 12092909 TI - Coping self-efficacy perceptions as a mediator between acute stress response and long-term distress following natural disasters. AB - The mediating effect of coping self-efficacy (CSE) perceptions between acute stress responses (ASR) and 1-year distress following two disasters was tested. Between 3 and 8 weeks after the second disaster and again at 1 year, 46 residents completed questionnaires. Posttraumatic Stress Disorder (PTSD) symptoms and global distress served as outcomes. Multiple regression demonstrated that ASR and Time I CSE were significant predictors of both Time 1 outcomes. Time 1 PTSD symptoms and Time 2 CSE were significant factors for Time 2 PTSD symptoms. Gender was significant for Time 2 PTSD symptoms, but not for Time 2 global distress. Longitudinally, Time 1 CSE predicted Time 2 PTSD symptoms, but not general distress. CSE mediated between ASR and both psychological outcomes at Time 2. Coping self-efficacy perceptions provide a possible intervention target. PMID- 12092910 TI - Sexual assault history and social support: six general population studies. AB - We evaluated the association of sexual assault history with later social support, operationalized as network size, marital status, presence of a partner, frequency of network contacts, and emotional support from friends and family, from spouse, and from partner. Data came from six independent general population surveys (pooled N = 9,865) whose results were summarized using meta-analysis. People who had been sexually assaulted were less likely than others to be married (OR = 0.75, 95% CI = 0.65, 0.87) or to report at least weekly contact with friends and relatives (OR = 0.48, 95% CI = 0.31, 0.75), and reported less emotional support from friends and family (OR = 0.72,95% CI = 0.58,0.89) and spouse (OR = 0.67, 95% CI = 0.54, 0.82). Results were consistent across studies, genders, and ethnic groups. Circumstances of sexual assault were sometimes related to social support. PMID- 12092911 TI - Emergency room vital signs and PTSD in a treatment seeking sample of motor vehicle accident survivors. AB - Seeking to replicate earlier reports by Shalev et al. (1998) and R. A. Bryant, A. G. Harvey, R. M. Guthrie, and M. L. Moulds (2000) that elevated heart rate (HR) shortly after a trauma was predictive of later posttraumatic stress disorder (PTSD), we examined vital sign data on 74 treatment-seeking motor vehicle accident (MVA) survivors, taken in the Emergency Department (ED) following their MVAs. Contrary to expectations, we found that those with elevated HRs in the ED were significantly less likely to meet criteria for PTSD 13 months post-MVA and that those with elevated HRs had lower levels of posttraumatic stress symptoms. Likewise, those with current PTSD had lower ED values of HR and DBP than did those who did not currently meet criteria for PTSD. PMID- 12092912 TI - Caregiver burden and psychological distress in partners of veterans with chronic posttraumatic stress disorder. AB - Caregiver burden and psychological distress were examined in a sample of 71 partners of Vietnam War combat veterans. Partners of patients (n = 51) diagnosed with posttraumatic stress disorder (PTSD) experienced more caregiver burden and had poorer psychological adjustment than did partners of veterans without PTSD (n = 20). Among PTSD caregivers, patient PTSD symptom severity and level of interpersonal violence were associated with increased caregiver burden. When accounting for patient PTSD symptom severity, hostility, presence of major depression, level of interpersonal violence, and health complaints, only PTSD severity was uniquely associated with caregiver burden. Caregiver sociodemographic factors including age, race, education, and the availability of social support, did not moderate the relationship between PTSD symptom severity and caregiver burden. Caregiver burden was strongly related to spouse psychological adjustment. PMID- 12092913 TI - Prevalence and correlates of secondary traumatic stress in workplace lay trauma counselors. AB - Quantitative (N = 130) and qualitative (N = 30) data were collected to explore the experiences of nonprofessional trauma counselors in the workplace. Counselors, on average, did not experience symptoms of secondary traumatic stress (STS) requiring clinical intervention. Changes to cognitive schemata regarding counselors' world views were found to be present 6 weeks after their last trauma counseling incident. Counselors reported experiencing considerable role satisfaction. Factors related to the nature of the trauma counseling task and counselors' exposure to work and nonwork related trauma were not significantly associated with counselors' STS or role satisfaction scores. Program coordination, self-efficacy, stakeholder commitment, sense of coherence, and perceived social support were significantly related to counselors' experiences of STS and role satisfaction. PMID- 12092914 TI - Shame and posttraumatic stress disorder. AB - Guilt about surviving a traumatic event is thought to be an associated feature of posttraumatic stress disorder (PTSD). Shame is an emotion closely related to guilt but is a distinct affective state. Little is known regarding the role of shame in PTSD and there are no studies of PTSD where shame and guilt are examined simultaneously. We used a measure of shame- and guilt-proneness in 107 community residing former prisoner of war veterans all of whom had been exposed to trauma. The measure of shame-proneness was positively correlated with PTSD symptom severity whereas guilt-proneness was not. This study provides the first empirical data regarding a possible role for shame in PTSD and may have important therapeutic and theoretical implications. PMID- 12092915 TI - Autobiographical memory specificity, intrusive memory, and general memory skills in Dutch-Indonesian survivors of the World War II era. AB - A lack of specific autobiographical memory may result from exposure to psychological trauma, intrusive memories of adverse events, and/or a general memory deficit. This study explored the role of these variables in 25 patients with various psychiatric diagnoses and 15 healthy controls. All participants had been exposed to war atrocities during their childhood in Indonesia. Patients produced significantly less specific memories than did controls. In addition, rather than performance on general neuropsychological memory tests, the frequent occurrence of intrusive memories and the avoidance of reminders of trauma predicted less autobiographical memory specificity. These results replicate and extend earlier findings on intrusive and nonspecific autobiographical memory in depressed samples to a group of people who experienced war atrocities in childhood. PMID- 12092916 TI - Parental caring as a possible buffer against sexual revictimization in young adult survivors of child sexual abuse. AB - This study examined whether parental caring provided a buffer against the revictimization effect. Nine hundred and seventy-four undergraduate women provided information about child sexual abuse, physical abuse, and whether they witnessed violence between their parents during childhood. They also reported whether they had ever been the victim of sexual assault in adulthood, and offered their perceptions of the degree of care they received as a child from each parent. Results indicated that women who had been sexually abused in childhood were twice as likely to be sexually assaulted in adulthood and that women with 2 or more types of childhood trauma were 3 times as likely to be sexually revictimized. Parental caring was not found to buffer against the revictimization effect. PMID- 12092917 TI - The relation of appraisal, coping efforts, and acuteness of trauma to PTS symptoms among former political prisoners. AB - We examined how trauma-specific appraisals and coping efforts mediate between traumatic experiences, acuteness of trauma, and length of imprisonment and posttraumatic stress symptoms (PTS) among 103 Palestinian former political prisoners. The findings provide support for both direct and mediated models of trauma. The acuteness of trauma (time since release), appraisal of prison experience as harmful and involving loss, and use of both emotion- and problem focused coping efforts were associated with high levels of PTS symptoms. Torture and ill-treatment had a direct association with intrusion, and recent release from prison with avoidance symptoms. Acuteness of trauma turned out to be important in the coping and symptom association: emotion-focused coping was associated with a low level of PTS symptoms in the long run, whereas problem focused coping was associated with a low level of PTS symptoms in the short run. PMID- 12092918 TI - Changes in PTSD patients' narratives during prolonged exposure therapy: a replication and extension. AB - Following E. B. Foa, C. Molnar, and L. Cashman (1995), narrative changes from the first to the last exposure session were compared for improved and nonimproved PTSD patients on fragmentation, organization, internal, and external events. Improved (n = 8) and nonimproved (n = 12) patients did not differ regarding changes in fragmentation or organized thoughts. However, improved patients showed a greater decrease in disorganized thoughts during treatment. Furthermore, all patients, independent of improvement, showed significant changes in the same direction; a decrease in disorganized thoughts and external events and an increase in internal events. Although previous results were partly replicated, it is concluded that narrative changes may be due to exposure treatment itself rather than to changes in memory representation. PMID- 12092919 TI - A prospective analysis of trauma exposure: the mediating role of PTSD symptomatology. AB - Trauma exposure has been associated with increased risk of exposure to additional traumatic events. Reactions to trauma exposure, specifically PTSD symptomatology, may mediate the link between trauma exposure and later traumatic events. Data from a longitudinal sample of Gulf War veterans (N = 2,949) were analyzed using a series of regression models. Higher levels of combat exposure were related to increased reports of PTSD symptomatology immediately upon return as well as increased reports of traumatic events in the 2 years following the Gulf War. PTSD symptomatology partially mediated the link between combat exposure and later trauma. Symptom clusters were also analyzed separately as potential mediators. Implications for the treatment of PTSD and prevention of exposure to multiple traumas are discussed. PMID- 12092920 TI - HIV-infected persons' knowledge of their sexual partners' HIV status. AB - The purpose of this study was to measure the accuracy of HIV-infected persons' knowledge of their sexual partners' HIV infection status. HIV-infected persons at two public health clinics reported their knowledge of the HIV infection status of their sexual partners. Actual HIV test results for sexual partners were then linked to survey responses. The association between reported knowledge and actual infection status of partners was estimated by kappa scores and percent agreement. Sixty-four percent (14/22) of partners thought to be infected were actually uninfected, and 42% (8/19) of partners thought to be uninfected were actually infected. The overall percent agreement was 46% (kappa = -0.06), less than that expected by random chance alone. Individuals in committed partnerships were less likely to have accurate knowledge than those in casual partnerships. In conclusion, HIV-infected individuals have poor knowledge regarding their partners' infection status. This may influence sexual behaviors that result in increased transmission. PMID- 12092921 TI - Prenatal HIV counseling and testing in California: women's experiences and providers' practices. AB - Two concurrent surveys were conducted in four California counties to compare the prenatal HIV counseling and voluntary testing (C&VT) experiences of women with the self-reported practices of prenatal care providers. Participants were 850 women currently or recently receiving prenatal care and 254 providers. Although 79.9% of women reported being offered an HIV test during a prenatal visit, only 56.2% said they were told about the risks and benefits of taking an HIV test. Almost all providers (98.4%) indicated they offer an HIV test, and 76.8% reported offering counseling, to every patient. One third of the women (65.9%) knew that treatment exists for reducing the chance of prenatal transmission of HIV, and 78.7% of women said they were more likely to be tested given knowledge of such therapy. Women may have underreported prenatal C&VT because providers spend insufficient time discussing related issues or because C&VT information is not presented in a way that is relevant to all patients. PMID- 12092922 TI - A model for capacity-building in AIDS prevention programs. AB - This article discusses a collaboratively organizational capacity-building model for agencies engaged in HIV/AIDS prevention. The cooperative training approach (CTA) utilized by the Training and Technical Assistance Program trained participants to incorporate learned skills into their daily work practices, and to become trainers to help disseminate their newly acquired skills, first to others in their own organization and later to other organizations. The CTA was successfully implemented among 285 community-based organizations (CBOs) serving predominantly Hispanics in the U.S.-Mexico border region. The program evaluation demonstrated that it has significantly upgraded the program implementation capability and the organizational infrastructure of the participating organizations resulting in improved delivery of services to their clients. Furthermore, it fostered networking and collaboration for the development of sustainable training and technical assistance networks of agency staff skilled in organizational capacity building. It is suggested that funding agencies examine the feasibility of using this approach to increase the organizational capacity of CBOs involved in HIV/AIDS prevention. PMID- 12092923 TI - Exploring the community response to a randomized controlled HIV/AIDS intervention trial in rural Uganda. AB - Investigators need, both for ethical and methodological reasons, to consider the acceptability of their intervention to the study population. We explored the response to a community-based randomized controlled trial (RCT) of an HIV/AIDS behavioral change intervention in rural Uganda. The views of field-workers, trial community, nongovernment organization representatives, and religious leaders were explored via focus groups (13) and interviews (45). The results suggest that the components of the intervention valued by the community are not necessarily those prioritised by trial implementers. Specifically, prevention activities appear to be valued less than material assistance. Furthermore, universal acceptance of the trial is probably unattainable. For these reasons, sensitive mobilization, respect for community members and their appointed leaders, and ongoing communication is essential. We suggest that evaluations of process be regarded as essential to the conduct of community-based RCTs and highlight the need for appropriate evaluation indicators to facilitate this. PMID- 12092924 TI - Reaching and identifying the STD/HIV risk of sex workers in Beijing. AB - China's HIV cases are growing by more than 30% annually. Few researchers have been able to reach sex workers and examine their risk behavior patterns. Key informants in illegal prostitution connected with owners of establishments in Beijing to recruit the involvement of sex workers. A total of 69 were surveyed about their background and risk behaviors in spring, 2000 in four venues; hair salons, bathing centers, karaoke bars, and the street. The women were mostly young (in their 20s), of low socioeconomic status, and from small towns and villages. They worked about 3 to 4 days a week, averaged one to two clients per day, made the equivalent of about U.S. $135 a week, and averaged about 3 months at any one establishment. They lacked accurate knowledge of STDs/HIV, and although 61% reported consistent condom usage 93% associated usage with prevention of pregnancy and 72% perceived a low risk of HIV infection. Nearly all (98%) felt condoms were affordable, yet 37% reported they were not accessible; 74% had obtained a medical exam in the last year, but 29% were examined when sick; only 10% received prevention information during medical visits, and 55% did not know where to go for HIV testing. Based on venue, stratification among sex workers existed, impacting their risk. Sex workers are at high risk of HIV infection/transmission, especially as the virus becomes more established in Beijing. Although risk factors were consistent with those of sex workers in other countries, the variation by venue, the low perceived vulnerability to HIV, the highly illegal nature of prostitution, and high mobility of sex workers in Beijing calls for a tailored intervention approach. Prevention messages/strategies should be responsive to the differing background, knowledge, PMID- 12092925 TI - Condom use among vocational school students in Chiang Rai, Thailand. AB - Condom use and its psychosocial correlates were investigated in a sample of 1,725 male and female vocational students (aged 15-21 years) in northern Thailand. Consistent condom use was relatively infrequent with all partner types (8.0% with recent steady partners, 28.5% with casual partners, and 30.7% with commercial sex partners), and only 24.3% reported condom use at first sex. These findings suggest that condom use, even with commercial partners, is not becoming widely established in the younger generation of Thai youths. Condom use with commercial partners was far below goals established by Thailand's 100% Condom Campaign, which was particularly significant for a population in Thailand's HIV epicenter. Consistent condom use with a steady partner was significantly related to condom use during first sex, which suggests the importance of establishing a "condom habit." Consistent condom use with casual partners was related to never having been pregnant (self or partner). Having used condoms at first sex also was associated with never having been pregnant (self or partner), as well as with a number of background social and psychological factors. Intentions to use condom were highly related to peer norms. Results suggest the importance of addressing peer norms concerning condom use, as well as the role of condoms in effective birth control. PMID- 12092926 TI - HIV/AIDS-related knowledge and attitudes: a comparison of older persons and young adults in Thailand. AB - Levels of AIDS related knowledge are widely assumed to be high in Thailand, a country with probably the most effective response to the HIV/AIDS pandemic to date in the developing world. But efforts to verify these levels are sparse in recent years, and very little attention has ever been paid to AIDS knowledge and attitudes within the Thai older population. Because many Thai older persons (aged 50 and over) remain sexually active late in life, and because many more will be involved in interactions with and care taking of young adult persons suffering from AIDS, we explore AIDS knowledge and attitudinal data we collected during 1999 from a sample of 773 older Thais from four provinces and Bangkok. The sample provinces were chosen to include a wide range of prevalence levels and social contexts regarding the epidemic. We compare the results with data from a sample of 398 young adults using the same questionnaire, collected at the same sites, at the same time (total N = 1,171). We find an overall high level of awareness about AIDS across our age groups of interest, but also identify important deficiencies among the young adults and especially among the older Thais. Implications of the findings are discussed. PMID- 12092927 TI - Education and HIV/AIDS prevention among young people. PMID- 12092928 TI - Evaluating HIV prevention efforts to reduce new infections and ensure accountability. PMID- 12092929 TI - Development and implementation of the cross-site evaluation of the CDC/HRSA corrections demonstration project. AB - U.S. prisons and jails have the nation's highest concentration of individuals infected with and at risk for HIV infection. Many correctional institutions offer 1-HIV care, but advances are oftentimes lost when individuals with a lack of health insurance and access to care are released into the community. In 1999 the Centers for Disease Control and Prevention and the Health Resources and Services Administration jointly funded seven health departments to address this need. These projects target soon-to-be-released inmates of jails, prisons, and juvenile facilities and offer enhanced discharge planning, case management, HIV prevention, disease screening, and staff training. The Evaluation and Program Support Center (EPSC) at Emory University and Abt Associates was funded to oversee a cross-site evaluation of these demonstration projects. This paper describes the process of developing a cross-site evaluation, the implementation of this evaluation, and lessons learned by the EPSC throughout this process. PMID- 12092930 TI - Gaps in technology transfer materials for HIV prevention program evaluation. PMID- 12092932 TI - HIV prevention evaluation: current contributions and future directions. PMID- 12092931 TI - A data warehouse concept for HIV prevention program evaluation. PMID- 12092933 TI - Designing and conducting participatory outcome evaluation of community-based organizations' HIV prevention programs. AB - With the increasing emphasis on evaluating the effectiveness of community-based organizations' HIV prevention programs, the needs, concerns, and strategies related to having stakeholders participate in designing and conducting an outcome evaluation need to be discussed. Stakeholders' participation in outcome evaluation ensures its relevancy and fairness. Participatory outcome evaluation starts with assessing the feasibility of conducting an outcome evaluation and determining whether stakeholders have a need for an outcome evaluation. If an outcome evaluation is possible and needed, the areas in which stakeholders can make important contributions to the evaluation are negotiated with the stakeholders. The article also discusses strategies to improve stakeholders' use of the results of outcome evaluation. PMID- 12092934 TI - Factors affecting the delivery of HIV/AIDS prevention programs by community-based organizations. AB - Community based organizations (CBOs) play a frontline role in HIV/AIDS prevention activities. CBOs face formidable challenges to effective delivery of HIV prevention services including client characteristics such as homelessness and CBO characteristics such as limited resources and staff turnover. Despite these obstacles, CBOs are generally well positioned to deliver services to specific high-risk populations because they understand their local communities and are connected to the groups they serve. [C1]This qualitative study illustrates that structural, sociocultural, organizational, and individual client factors both facilitate and act as barriers to delivery of HIV prevention services. These challenges and successes help identify critical technical assistance needs. PMID- 12092936 TI - Lessons learned from the first year of implementation of the Centers for Disease Control and Prevention's standardized evaluation system for HIV prevention programs. AB - In December 1999 the Centers for Disease Control and Prevention's (CDC's) Division of HIV/AIDS Prevention initiated a standardized evaluation system for CDC-funded health department HIV prevention programs. This health department evaluation guidance asks health departments to develop comprehensive evaluation plans and to submit aggregated data on such activities as intervention planning, process monitoring, and outcome evaluation. During the first year of this system, of 65 health departments, 62 submitted evaluation plans, 37 submitted intervention plan data, and 20 submitted process monitoring data. Major issues affecting implementation of a national evaluation system include varying levels of evaluation capacity among health departments, differences between the CDC's taxonomy for national data collection and local definitions, and limitations regarding use of 1st-year data. The CDC has learned that implementation of a standardized evaluation system takes considerable time and that stakeholder involvement and technical assistance and capacity building support are essential. PMID- 12092935 TI - Evaluation barriers and facilitators among community-based HIV prevention programs. AB - Funding agencies are using technical assistance to promote evaluation of their community-based HIV prevention programs. Using qualitative methods, we identified 11 factors that hinder and facilitate evaluation within community-based organizations (CBOs): staff perceptions, availability of funding, data collection, data validity, data utility, technical assistance, effects on services, effects on funding, staff skills, tools and technology, and expectations of the funding agency. Using these factors, we developed eight strategies to promote CBO evaluation. These strategies go beyond the traditional role of technical assistance and address the broader context within which CBOs evaluate their programs. Funding agencies and technical assistance providers can use these strategies to enhance CBO evaluation, which may result in more and better quality evaluations and, ultimately, improvements in the effectiveness of HIV prevention services. PMID- 12092937 TI - The value of engaging stakeholders in planning and implementing evaluations. AB - Stakeholder participation in evaluation has surfaced as a major factor contributing to the effectiveness of HIV prevention programs. In recognition of the multiple benefits, the Centers for Disease Control and Prevention (CDC), has used a framework to involve stakeholders in the evaluation of its programs. This article describes the framework used by the CDC and provides examples of four studies that involved various stakeholders from health departments, community based organizations, and community planning groups to national and regional organizations in designing and implementing evaluations that yielded results useful for program improvement. The participatory process involved stakeholders in each of the four phases of the framework: evaluation planning, implementation, development of action plans, and dissemination. Lessons learned include the importance of having a facilitator to coordinate activities and ongoing communication with those involved in the evaluation. Stakeholders shared that using the evaluation results for action planning was beneficial for improving their programs. Despite many challenges faced in the stakeholder evaluation process, most stakeholders agreed that many benefits grew out of the multiple perspectives presented and understanding of the service agencies. PMID- 12092938 TI - A randomized controlled intervention trial of a sexual health approach to long term HIV risk reduction for men who have sex with men: effects of the intervention on unsafe sexual behavior. AB - This controlled prospective study assessed the effectiveness of a sexual health approach to HIV prevention for men who have sex with men (MSM). Participants (N = 422 Midwestern MSM) were randomly assigned to the intervention group, who participated in a 2-day comprehensive human sexuality seminar designed to contextually address long-term risk factors and cofactors, or to the control group, who watched 3 hours of HIV prevention videos. Risk behavior during the preceding 3 months was measured at baseline, 3-month follow-up, and 12-month follow-up. Any unprotected anal intercourse outside a long-term seroconcordant relationship was the dependent variable. Of the total, 14%-24% of the participants were considered at risk of acquiring or transmitting HIV. At the 12 month follow-up, the control reported a 29% decrease in the use of condoms during anal intercourse; the intervention group reported an 8% increase (t = 2.546; p = .015). The sexual health seminars appear a promising new intervention at significantly reducing unprotected anal intercourse between men. PMID- 12092939 TI - Evaluation of HIV prevention and counseling practices of obstetrician/gynecologists in Bogota, Colombia: impact on women's knowledge and risk practices. AB - The present community-based study in Bogota, Colombia, investigated risk assessment and preventive counseling practices of obstetrician/gynecologists (ob/gyn n = 34) and their impact on women's (n = 230) knowledge and risk behaviors. The data indicate that physician education has a significant and positive impact on women's knowledge and behavior. After controlling for sociodemographic variables, women instructed by their ob/gyn were 11 times more likely to correctly identify preventive measures (p = 0.0001) and high-risk sexual practices for the HIV /sexually transmitted diseases (STDs) transmission (4x; p = 0.05) and were less likely to engage in high-risk sexual practices (OR = 2; p = 0.05). Few ob/gyns (17%), however, assess risk behaviors and provide risk reduction counseling, and only 6% frequently encouraged HIV testing. These findings highlight the potential public health impact of ob/gyn physicians and underscore the need to increase their involvement in halting the HIV/STD epidemic in Colombia. PMID- 12092940 TI - Evaluation of a sexual health approach to prevent HIV among low income, urban, primarily African American women: results of a randomized controlled trial. AB - This randomized controlled trial evaluated an innovative culturally specific sexual health intervention-targeting, but not limited to, low-income African American women-in which HIV and sexually transmitted disease prevention strategies were combined with comprehensive sexuality education. The intervention was delivered and evaluated in community-based settings to 218 participants randomly assigned to treatment or a no-treatment control group. Participants were interviewed at pretest and 3 and 9 months after the intervention to assess changes in both sexuality and HIV risk variables. The intervention was effective in improving sexual anatomy knowledge at both 3- and 9-month follow-up. For a subset of women engaging in unprotected sex at pretest, the intervention group reported an increase in positive attitudes toward the female condom at 9-month follow-up. Reasons for the weak treatment effect are discussed in the context of challenges inherent in conducting community-based research with high-risk populations and sensitive topics. Recommendations are provided for improving sample attrition, statistical power, and response bias and for altering the intervention so as to strengthen its impact. PMID- 12092941 TI - Rapidly changing conditions in the brothels of Bangladesh: impact on HIV/STD. AB - Bangladesh is a low HIV prevalence country with several well-documented at-risk groups, the most prominent of which is brothel-based sex workers. Using two waves of HIV behavioral surveillance data for a national sample of all operating registered brothels supplemented by historical and observational accounts, this article presents a case study of the changing conditions in the brothels. Between the two waves of surveillance, several brothels were forcibly closed; the number of sex workers dropped; the average number of clients per woman rose; and overall safety, both in terms of violence and protected sex, did not improve. Only treatment-seeking behavior for sexually transmitted diseases was positively changed. Continued surveillance of the possible negative impact of HIV prevention programs is under way. Protective policies are needed to facilitate improved HIV prevention and safety of sex workers, a key to controlling the HIV epidemic. PMID- 12092942 TI - Farm bill includes animal health, welfare provisions promoted by AVMA. Congress addresses research animals,TSEs, and nonambulatory livestock. American Veterinary Medical Association. PMID- 12092943 TI - Wisconsin mobilizes to battle chronic wasting disease. PMID- 12092944 TI - BVDV: 40 years of effort and the disease still has a firm hold. PMID- 12092945 TI - Sees limitations of clinical reports. PMID- 12092946 TI - Commends successful use of electrical cardioversion in a horse. PMID- 12092947 TI - More on the pregnant mare urine industry. PMID- 12092948 TI - Grateful for article analyzing women in veterinary medicine. PMID- 12092949 TI - What is your diagnosis? Distension of the stomach with gas and ingesta. PMID- 12092950 TI - Perceptions of state public health officers and state veterinarians regarding risks of bioterrorism in the United States. AB - OBJECTIVE: To assess perceptions of state public health officers and state veterinarians in the United States regarding the risks of bioterrorism and determine the degree of support provided for activities related to bioterrorism. DESIGN: Cross-sectional survey. SAMPLE POPULATION: State public health officers and state veterinarians. PROCEDURE: A questionnaire was sent between April and June 2001 to the state public health officer and state veterinarian in each of the 50 states and the District of Columbia. RESULTS: Perceptions of the risk of bioterrorism attacks were similar for state public health officers and state veterinarians. Veterinarians perceived the risks associated with foot-and-mouth disease and Newcastle disease to be higher than did physicians. State veterinarians perceived the risks associated with an anthrax hoax, brucellosis, and ricin toxicosis to be lower than did state public health officers. Risk posed by agents that affected animals exclusively was perceived to be higher than risk posed by agents that affected humans exclusively and zoonotic agents. Number of full-time-equivalent positions devoted to bioterrorism surveillance and percentage of the budget devoted to bioterrorism activities were significantly lower for offices run by state veterinarians than for offices run by state public health officers. State veterinarians were significantly less likely to have knowledge of bioterrorism incidents within their state or district than were state public health officers. CONCLUSIONS AND CLINICAL RELEVANCE: Provision of additional resources to state veterinarians and explicit integration of their expertise and surveillance capabilities may be important to effectively mitigate the risk of bioterrorism. PMID- 12092951 TI - Prevalence of systolic hypertension in cats with chronic renal failure at initial evaluation. AB - OBJECTIVE: To determine prevalence of systolic hypertension and associated risk factors in cats with chronic renal failure evaluated in first-opinion practice. DESIGN: Prospective study. ANIMALS: 103 cats with chronic renal failure. PROCEDURE: Systolic arterial blood pressure (SABP) was measured with a noninvasive Doppler technique, and cats that had SABP > 175 mm Hg on 2 occasions or that had SABP > 175 mm Hg and compatible ocular lesions were classified as hypertensive. Information from the history (previous treatment for hyperthyroidism, age), physical examination (sex, body weight), routine plasma biochemical analyses (creatinine, cholesterol, potassium, sodium, chloride, and calcium concentrations), and thyroid status were evaluated as potential risk factors for systolic hypertension. Variables associated with systolic hypertension were evaluated by use of logistic regression. RESULTS: 20 (19.4%; 95% confidence interval, 13 to 28%) cats had systolic hypertension. Plasma potassium concentration was significantly and inversely associated with systolic hypertension. CONCLUSIONS AND CLINICAL RELEVANCE: Prevalence of systolic hypertension, although clinically important, was lower than that reported previously. The cause of the inverse association between systolic hypertension and plasma potassium concentration is not yet known. PMID- 12092952 TI - Evaluation of a manual technique for detection of neutropenia and thrombocytopenia in dogs receiving chemotherapy. AB - OBJECTIVE: To determine accuracy of a manual technique for detection of neutropenia and thrombocytopenia in dogs receiving chemotherapy. DESIGN: Masked prospective study. ANIMALS: 11 dogs treated with chemotherapy for neoplasia. PROCEDURE: 124 blood samples from dogs being treated with chemotherapy for various neoplasms were processed through an automated cell counter, and results were compared with those obtained by use of a rapid manual technique for estimating neutrophil and platelet concentrations to determine whether the manual technique could accurately detect dogs with neutropenia or thrombocytopenia. RESULTS: By use of automated techniques, neutropenia (< 3,000 cells/microl) was detected in 17 of 124 blood samples, and thrombocytopenia (< 100,000 platelets/microl) was detected in 3 of 124 blood samples. The manual technique correctly identified 16 of 17 (94%) blood samples with neutropenia, with a specificity of 92% (98/107). The manual technique correctly identified 3 of 3 (100%) blood samples with thrombocytopenia, with specificity of 94% (114/121). CONCLUSIONS AND CLINICAL RELEVANCE: Manual estimates of neutrophil and platelet counts are sensitive and specific; however, a full differential cell count is still preferable. PMID- 12092953 TI - Comparison of pulse administration versus once daily administration of itraconazole for the treatment of Malassezia pachydermatis dermatitis and otitis in dogs. AB - OBJECTIVE To compare clinical efficacy of pulse administration with itraconazole versus once daily administration for the treatment of cutaneous and otic M pachydermatis infection in dogs. DESIGN: Randomized controlled trial. ANIMALS: 20 dogs. PROCEDURE: Dogs were treated with itraconazole orally (n = 10/group), using a pulse administration regimen (5 mg/kg [2.3 mg/lb], PO, q 24 h for 2 consecutive days per week for 3 weeks) or once daily administration (5 mg/kg, PO, q 24 h for 21 days). No other treatment was permitted. On days 0 and 21, clinical severity of cutaneous and otic disease was assessed, and samples were collected for cytologic examination and yeast culture. Cytology (sum of the mean number of yeast organisms per oil immersion field for affected sites) and culture (mean of the score for extent of yeast growth for samples from affected sites) scores were calculated. RESULTS: For dogs in both treatment groups, clinical severity of cutaneous and otic disease was significantly decreased by day 21, but decrease in severity was not significantly different between groups. Similarly, skin cytology, skin culture, and ear culture scores were significantly decreased on day 21, compared with day 0, for both groups, but decreases were not significantly different between groups except that dogs in the pulse administration group had a significantly greater decrease in ear culture scores than did dogs in the daily administration group. However, when cytology scores only for ear samples were analyzed, day 21 score was not significantly decreased, compared with day 0 score, for either group. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that both pulse administration and once daily administration of itraconazole were efficacious in the treatment of M pachydermatis cutaneous infection in dogs. However, adjunctive treatment may be needed in dogs with M pachydermatis otitis. PMID- 12092954 TI - Evaluation of treatment with doxorubicin and piroxicam or doxorubicin alone for multicentric lymphoma in dogs. AB - OBJECTIVE: To evaluate the antitumor and toxic effects of treatment with doxorubicin combined with piroxicam or doxorubicin alone for multicentric lymphoma in dogs. DESIGN: Nonrandomized clinical trial. ANIMALS: 75 dogs with multicentric lymphoma. PROCEDURE: 33 dogs were treated with doxorubicin (30 mg/m2, IV, q 21 d, for 3 doses) and piroxicam (0.3 mg/kg [0.14 mg/lb], PO, q 24 h); results were compared with a historical control group of 42 dogs treated with doxorubicin (30 mg/M2, IV, q 21 d, for 3 doses) alone. Results-The percentages of dogs that had remission with doxorubicin-piroxicam treatment (79%) or doxorubicin treatment alone (74%) were not significantly different. Median duration of first remission was 130 days with doxorubicin-piroxicam and 147 days with doxorubicin alone; these values were not significantly different. Severe toxicosis was observed in 22% of dogs treated with doxorubicin-piroxicam and 17% of dogs treated with doxorubicin alone. CONCLUSIONS AND CLINICAL RELEVANCE: Both treatment protocols were efficacious and well tolerated. The doxorubicin piroxicam treatment was no more effective regarding response rate, remission duration, or survival duration, compared with the control group treated with doxorubicin alone. PMID- 12092955 TI - Effects of preoperative administration of ketoprofen on whole blood platelet aggregation, buccal mucosal bleeding time, and hematologic indices in dogs undergoing elective ovariohysterectomy. AB - OBJECTIVE: To determine effects of preoperative administration of ketoprofen on whole blood platelet aggregation, buccal mucosal bleeding time, and hematologic indices in dogs after elective ovariohysterectomy. DESIGN: Randomized, masked clinical trial. ANIMALS: 22 healthy dogs. PROCEDURE: 60 minutes before induction of anesthesia, 11 dogs were given 0.9% NaCl solution (control), and 11 dogs were given ketoprofen (2 mg/kg [0.9 mg/lb], IM). Thirty minutes before induction of anesthesia, glycopyrrolate (0.01mg/kg [0.005 mg/lb]), acepromazine (0.05 mg/kg [0.02 mg/lb]), and butorphanol (0.2 mg/kg 10.09 mg/lb]) were given IM to all dogs. Anesthesia was induced with thiopental (5 to 10 mg/kg [2.3 to 4.5 mg/lb], IV) and maintained with isoflurane (1 to 3%). Ovariohysterectomy was performed and butorphanol (0.1 mg/kg [0.05 mg/lb], IV) was given 15 minutes before completion of surgery. Blood samples for measurement of variables were collected at intervals before and after surgery. RESULTS: In dogs given ketoprofen, platelet aggregation was decreased 95 +/- 10% and 80 +/- 35% (mean +/- SD) immediately after surgery and 24 hours after surgery, respectively, compared with preoperative values. At both times, mean values in dogs given ketoprofen differed significantly from those in control dogs. Significant differences between groups were not observed for mucosal bleeding time or hematologic indices. CONCLUSIONS AND CLINICAL RELEVANCE: Preoperative administration of ketoprofen inhibited platelet aggre gation but did not alter bleeding time. Ketoprofen can be given before surgery to healthy dogs undergoing elective ovariohysterectomy, provided that dogs are screened for potential bleeding problems before surgery and monitored closely after surgery. PMID- 12092956 TI - Tenoscopic-assisted treatment of open digital flexor tendon sheath injuries in horses: 20 cases (1992-2001). AB - OBJECTIVE: To determine history, physical and diagnostic examination findings, medical treatment, and outcome of horses with open injuries to the digital flexor tendon sheath treated with the assistance of tenoscopy. DESIGN: Betrospective study. ANIMALS: 20 horses. PROCEDURE: Medical records of 20 horses with open injuries to the digital flexor tendon sheath were reviewed. Signalment, history, physical and diagnostic examination results, bacteriologic culture and susceptibility testing results, surgical and medical treatments, and follow-up examination results were determined. Outcome was determined by use of telephone interview or physical examination. RESULTS: All horses were treated with tenoscopic-assisted lavage and debridement. Eighteen horses survived, and 2 were euthanatized during treatment. All horses were either grade-4 or grade-5 lame before treatment. Ten horses returned to previous use. Four horses were considered mildly lame and in athletic use. Three horses were considered mechanically lame and are in use with reduced expectations. One horse was lost to follow-up after being sold. One horse was euthanatized for financial reasons and 1 because of complications from regional sepsis. CONCLUSIONS AND CLINICAL RELEVANCE: Tenoscopy appears to be a useful modality in the treatment of open injury to the digital flexor tendon sheath in horses. Direct viewing, guided debridement, and targeted large-volume lavage are advantages obtained with intrathecal arthroscopy. Tenoscopy, when combined with antimicrobial and anti inflammatory treatment, appears to offer a good chance of survival for affected horses. PMID- 12092957 TI - Evaluation of early fetal losses on four equine farms in central Kentucky: 73 cases (2001). AB - OBJECTIVE: To determine features of an early fetal loss (EFL) syndrome and evaluate potential risk factors for EFL in Thoroughbred broodmares on 4 farms in central Kentucky. DESIGN: Retrospective study. ANIMALS: 288 pregnant broodmares. PROCEDURE: Year-2001 breeding records for 288 Thoroughbred broodmares were examined. Early fetal loss was defined as loss of a fetus that was viable at > or = 40 days of gestation but was subsequently lost by 5 months of gestation. RESULTS: Overall 2001 EFL rate was 25% (73/288), median gestational age at time of fetal loss was 77 days, and median date of loss was May 7. Mares on 1 farm had significantly fewer fetal losses (5%) than mares on the other 3 farms (26 to 35%). Fetal losses were higher for maiden (42%) and barren (42%) mares than for foaling mares (18%). Fetal losses were greater in young than in older mares. Effects of broodmare farm, mare age, and reproductive status were all significant. Fetal losses were not associated with sire used for mating or stud farm. CONCLUSIONS AND CLINICAL RELEVANCE: Greatest risk for EFL occurred during the period from late April to May (ie, in mares bred during February through March). Higher incidence of EFL in maiden and barren mares and lower incidence of EFL on 1 farm suggest management or environmental influences may have affected outcome. Risk factors that should be investigated include environmental differences among farms and differences in management procedures used for lactating versus nonlactating mares. PMID- 12092959 TI - Use of a parainguinal approach for cryptorchidectomy in alpacas. AB - Cryptorchidism is an uncommon condition in llamas and alpacas, and there are no reports detailing surgical management of this condition in llamas or alpacas; however, flank or ventral midline approaches have been mentioned. Cryptorchid castration was performed by use of a parainguinal approach in 2 alpacas. The retained testicle of each alpaca was found on the ventral aspect of the abdomen, just caudal to the incision in 1 alpaca and at the cranial edge of the incision in the other. The testicle was approximately 1 X 0.5 cm in 1 alpaca and 2 X 1 cm in the other. In an alpaca or llama with a single abdominal testicle and no history of surgery, the parainguinal approach has several advantages, including the likely proximity of the testicle to the incision; the small incision, which can be extended if needed; ease of closure; and minimal aftercare. The retained testicles were small and could be difficult to find from a more distant midline, paramedian, or flank incision. PMID- 12092958 TI - Urethroscopy and laser lithotripsy for the diagnosis and treatment of obstructive urolithiasis in goats and pot-bellied pigs. AB - OBJECTIVE: To explore the use of urethral endoscopy and laser lithotripsy in the diagnosis and management of urolithiasis in goats and pot-bellied pigs. DESIGN: Prospective clinical study. ANIMALS: 16 male goats and 6 male pot-bellied pigs with dysuria. PROCEDURE: Abdominal ultrasonography and urethral endoscopy were performed on all 22 animals. Endoscopic-guided holmium:yttrium-aluminum-garnet laser lithotripsy was performed in 3 goats and 2 pot-bellied pigs. RESULTS: Urolithiasis was identified in 15 goats and 5 pot-bellied pigs. Primary urinary bladder paralysis and cystitis were identified in the remaining pot-bellied pig and goat. Mean bladder diameters of obstructed small- and large-breed goats were 7 and 9.5 cm, respectively. The mean bladder diameter of obstructed pot-bellied pigs was 9.5 cm. Five of 20 animals with obstructive urolithiasis had severe urethral necrosis or stricture formation at the time of urethroscopy. All of these animals were euthanatized within 6 months because of persistent dysuria. When used, laser lithotripsy successfully fractured the distally located obstructing stones in the 3 goats and 2 pot-bellied pigs. CONCLUSIONS AND CLINICAL RELEVANCE: Urethral endoscopy is useful for evaluating urethral patency in goats and pot-bellied pigs. Examination of the urethral mucosa following relief of urethral obstructions aids in the assessment of the long-term prognosis for urethral stricture. Urethral endoscopy also expands the therapeutic options for management of urolithiasis by providing a route for conducting laser lithotripsy. Laser lithotripsy proved to be safe and effective for clearing distally located calculi refractory to removal by traditional urethral flushing. Lithotripsy application is restricted to calculi lodged in the urethra. PMID- 12092960 TI - Honey mesquite toxicosis in a goat. AB - Honey mesquite (Prosopis glandulosa) is distributed across a large portion of the southwestern United States. Ingestion of young leaves, pods, or beans can cause toxicosis in cattle and goats if they comprise a substantial portion of their diet. Goats, as browsers, are most likely to develolp mesquite toxicosis. Sheep appear to be more resistant to the plant's toxic effects. Consistent clinical signs include weight loss, ptyalism, mandibular tremors, tongue protrusion, and dysphagia. Diagnosis of mesquite toxicosis is largely made on the basis of history and clinical signs with exclusion of appropriate differentials. Laboratory findings are nonspecific but may reveal a mild anemia and hypoglycemia. Postmortem findings suggestive of mesquite toxicosis are limited to fine vacuolation of neurons in the trigeminal motor nucleus. Treatment consists of an alternative diet and supportive care. The disease is treatable in cattle and sheep but has a high case fatality rate in goats. PMID- 12092961 TI - An analysis of marine animal injuries presenting to emergency departments in Victoria, Australia. AB - OBJECTIVE: To describe the epidemiology of marine animal injury in Victoria, Australia, in order to identify risk factors and recommend prevention strategies. METHODS: Retrospective, descriptive study of patients with marine animal injuries who presented to Victorian emergency departments between October 1995 and June 2000. Data were obtained from the Victorian Emergency Minimum Dataset. The main outcome measures were the marine animal involved; the nature, time, and place of injury; and subject demographics and activity. RESULTS: Two hundred five injuries were identified, and males predominated (71.7%, P < .01). Injuries were most frequent during summer and when jellyfish were most prevalent. Various fish species, stingrays, jellyfish, and sharks were incriminated in 83 (40.5%), 46 (22.4%), 42 (20.5%), and 5 (2.4%) injuries, respectively. Most (65.9%) injuries occurred during leisure or sport, and 72 (35.1%) occurred in a place of recreation. Spikes, spines, and barbs caused 82 (40.0%) injuries, and stings caused 54 (26.3%) injuries. Bites were uncommon. Most injuries were to the limbs, with the hands or feet injured in 127 (62.0%) patients. Forty (19.5%) injuries were associated with a retained foreign body. Only 17 (8.3%) patients required admission to the hospital. CONCLUSIONS: Marine animal injury is seasonal but rarely serious. Vigilance is required when handling fish, and protective gloves, footwear, and clothing are recommended where appropriate. Clinicians should consider retained foreign bodies in penetrating injuries. Warnings are recommended when jellyfish are most prevalent. PMID- 12092962 TI - An analysis of human pathogens found in horse/mule manure along the John Muir Trail in Kings Canyon and Sequoia and Yosemite National Parks. AB - OBJECTIVE: To determine the prevalence of microorganisms that are potentially pathogenic for humans in horse/mule manure along the John Muir Trail (JMT). METHODS: Random samples of horse/mule manure were collected along sections of the JMT in Yosemite, Kings Canyon, and Sequoia national parks (NP), as well as in portions of the Pacific Crest Trail (PCT) and selected JMT/PCT access trails. Convenience samples of wild animal scat found within I mile of trails were also collected. The fresh specimens were individually preserved both in 0.9% saline and polyvinyl alcohol (PVA)-containing tubes and stored at 4 degrees C until time of analysis. Bacteriological analysis was performed using standard microbiology laboratory procedures. PVA samples were stained with trichrome and were then examined by a parasitologist. RESULTS: Collection: A total of 186 trail miles were sampled, including 113 on the JMT (Yosemite 37, Kings 53, and Sequoia 23). The PCT samplings included 24 miles, and NP and wilderness area access trails added an additional 49 miles. A total of 102 samples were collected, which included 81 samples from pack animals and 21 identified as having come from wild animals. Pack Animal Bacteria: All plated specimens grew large numbers of commensal gut flora. Potential pathogenic bacteria were found in only 12 samples and included Hafnia alvei (4), Serratia odorifera (1), Citrobacter freundii (1), Escherichia vulneris (1), Clostridium clostridioforme (1), Yersinia enterocolitica (1), Sherwinella putraformus (1), and Enterobacter spp (4). No Escherichia coli O157, Salmonella, or Aeromonas were found. Microscopic examination for protozoal organisms revealed occasional commensal ciliates and I Giardia. Wild Animal Pathogens: One specimen grew Y enterocolitica, and another grew Enterobacter amnigenus. CONCLUSIONS: We found a low prevalence of human pathogens in pack animal manure on the JMT. PMID- 12092963 TI - Year 2000 Whitewater Injury Survey. AB - OBJECTIVE: To gather retrospective data on paddling style, equipment, and acute and chronic/overuse injuries in hard shell whitewater canoe and kayak paddlers, to compare injury rates and patterns in different groups of paddlers, and to evaluate the incidence in paddlers of self-reported giardia infection. METHODS: In the summer and fall of 2000, a survey was distributed at riverside, through paddle club bulletins, and was posted on the Internet. RESULTS: Three hundred nineteen useable surveys were returned reporting 388 acute and 285 chronic injuries. The shoulder, wrist/hand, and elbow/forearm were the most common sites of injury. Sprain/strain was the most common (26%) known acute injury, followed by laceration and contusion (each 17%). Tendinitis was the most common (44%) known chronic injury diagnosis, followed by sprain/strain (27%). Forty-seven percent of acute and 36% of chronically injured paddlers sought medical attention. Giardia infection was reported in 14%. CONCLUSIONS: The shoulder and wrist/hand areas were the most common injured sites in this survey. Sprains, tendinitis, lacerations, and contusions were the most common known injury diagnoses. Injuries due to portage were common. Giardia infection may be common in whitewater paddlers. PMID- 12092964 TI - A case report of hypothermia in the wilderness. AB - Accidental hypothermia due to environmental exposure is a common condition encountered in the wilderness setting. I report a case of hypothermia that occurred in New Hampshire to a 42-year-old, poorly prepared mountaineer. He presented with ataxia, impaired judgment, oliguria, and slight confusion, suggestive of mild to moderate hypothermia. He was treated in the field using passive and active external rewarming methods and 1 active internal method. The signs, symptoms, and progression of hypothermia are reviewed, and treatment options are discussed, with an emphasis on management of hypothermia in the wilderness setting. Additionally, more complicated treatment modalities are canvassed, as these are of use to rescue teams and expeditions. Current controversies regarding the treatment of hypothermia, resuscitation efforts, and potential complications are also mentioned. PMID- 12092965 TI - Pleural tuberculosis in a Nepali trekker. PMID- 12092966 TI - Illness and injury among long-distance hikers on the Long Trail, Vermont. PMID- 12092967 TI - Respiratory function changes in a wilderness multisport endurance competition: a prospective case study. AB - OBJECTIVE: To document serially and in detail the changes in respiratory function during competition in a typical wilderness multisport endurance athlete. METHODS: Prospective observational case study over two years during a 105 km wilderness multisport endurance competition consisting of cross country skiing, cross country running, cycling, and whitewater canoeing. Nedocromil prophylaxis was used during the second year of the study only. FEV1, FVC, flow volume loops, and oxygen saturation were measured serially during the race. RESULTS: Baseline spirometry and histamine bronchial provocation tests were normal. SaO2% decreased to a minimum of 92%. Decreases in both FEV1 (up to 25%) and FVC (up to 22%) were observed at race finish in both years. Although FEV1/FVC ratios only decreased marginally, the pattern of change in maximum expiratory flow rates was most consistent with evolving airflow obstruction. The changes were not prevented by nedocromil. The subject did not develop symptomatic wheezing or abnormal breathlessness and finished the event without incident. CONCLUSIONS: Ultra endurance multisport competition may induce substantial changes in respiratory function. These appear to be attributable to evolving airflow obstruction. PMID- 12092968 TI - Sudden unexpected death on challenge courses. PMID- 12092969 TI - Technological advances in avalanche survival. AB - Over the last decade, a proliferation of interest has emerged in the area of avalanche survival, yielding both an improved understanding of the pathophysiology of death after avalanche burial and technological advances in the development of survival equipment. The dismal survival statistics born out of the modern era of winter recreation unmistakably reveal that elapsed time and depth of burial are the most critical variables of survival and the focus of newer survival devices on the market. Although blunt trauma may kill up to one third of avalanche victims, early asphyxiation is the predominant mechanism of death, and hypothermia is rare. A survival plateau or delay in asphyxiation may be seen in those buried in respiratory communication with an air pocket until a critical accumulation of CO2 or an ice lens develops. The newest survival devices available for adjunctive protection, along with a transceiver and shovel, are the artificial air pocket device (AvaLung), the avalanche air bag system (ABS), and the Avalanche Ball. The artificial air pocket prolongs adequate respiration during snow burial and may improve survival by delaying asphyxiation. The ABS, which forces the wearer to the surface of the avalanche debris by inverse segregation to help prevent burial, has been in use in Europe for the last 10 years with an impressive track record. Finally, the Avalanche Ball is a visual locator device in the form of a spring-loaded ball attached to a tether, which is released from a fanny pack by a rip cord. Despite the excitement surrounding these novel technologies, avalanche avoidance through knowledge and conservative judgment will always be the mainstay of avalanche survival, never to be replaced by any device. PMID- 12092970 TI - Clinical images. Deep lacerations to both hands. PMID- 12092971 TI - Death from lightning. 1872. PMID- 12092972 TI - Unusual injuries caused by lightning stroke. 1894. PMID- 12092973 TI - Tuckerman gothic. PMID- 12092974 TI - The "knot method" of tick removal. PMID- 12092975 TI - Death of unknown origin in alpine medicine. PMID- 12092976 TI - Personal experience with jack o'lantern mushroom toxicity. PMID- 12092977 TI - Simulated field experience in the use of the Sam splint for pressure immobilization of snakebite. PMID- 12092978 TI - Oxidative stress in humans training in a cold, moderate altitude environment and their response to a phytochemical antioxidant supplement. AB - OBJECTIVE: This study examined the effectiveness of an antioxidant mixture containing vitamin E, beta-carotene, ascorbic acid, selenium, alpha-lipoic acid, N-acetyl 1-cysteine, catechin, lutein, and lycopene to reduce oxidative stress in US Marines undergoing 24 days of cold-weather field training at a moderate altitude. METHODS: Forty physically active male volunteers (ages 18-40) were randomly assigned to a treatment (antioxidant) group (n = 21) or a control (placebo) group (n = 19). Breath pentane (BP), serum lipid hydroperoxides (LPO), urine malondialdehyde (MDA), urine 8-hydroxy deoxyguanosine (8-OHdG), ferric reducing ability of plasma (FRAP), and serum and urine oxygen radical absorption capacity (ORAC) were measured as indicators of oxidative stress and antioxidant status. Urine was sampled at days 0, 12, and 24. Serum and breath were sampled on days 0 and 24. RESULTS: Both groups exhibited increased levels of oxidative stress after 24 days of field training, as indicated by an increased LPO, pentane, and 8-OHdG. There was no significant difference between the treatment and placebo groups at day 24; however, there was some indication that test subjects with initially low antioxidant capacity (ORAC) may have benefited from the antioxidant supplement. CONCLUSIONS: An increased level of oxidative stress was associated with high levels of physical exertion of training in a cold environment at moderate altitude. The antioxidant mixture tested did not attenuate the mean oxidative stress levels in the entire group of test subjects, but it may have reduced the oxidative stress of some individuals with low initial antioxidant status. PMID- 12092979 TI - Integrated monitoring of a new group B streptococcal disease prevention program and other perinatal infections. AB - OBJECTIVE: To determine levels of prenatal screening for several infections, intrapartum recognition of risk factors, and prophylaxis against mother-to-child transmission of group B streptococcus. METHODS: Review of stratified random sample of hospital records for deliveries in Connecticut during 1996. SUDAAN analysis was used to adjust for the complex survey design, and weighting adjusted for the probability of being sampled and nonresponse. RESULTS: Of 992 records requested, 868 (88%) were abstracted and analyzed. Thirty-six percent of women had prenatal screening for group B streptococcus and 26% had been tested for human immunodeficiency virus (HIV), while 97-99% of women had been screened prenatally for hepatitis B surface antigen, rubella, and syphilis. Of those women tested, 17% were detected as group B streptococcus carriers, and 78% of these received intrapartum antibiotic prophylaxis. Among women who were not screened for group B streptococcus prenatally, 22% met risk-based criteria for prophylaxis, but only 45% of these received intrapartum prophylaxis. Among unscreened women with a risk factor, those with shorter hospital stays prior to delivery, admitted on evening or night shifts, or who delivered on the weekend were significantly less likely to receive intrapartum prophylaxis. CONCLUSION: In 1996, the majority of women who delivered in Connecticut were not tested prenatally for group B streptococcus and the majority of those not tested in whom there was an indication for prophylaxis were not treated. Compliance with group B streptococcus prevention recommendations can be improved through increased prenatal testing and/or better recognition of risk-based criteria for intrapartum prophylaxis. PMID- 12092980 TI - Assessing capacity and measuring performance in maternal and child health. AB - OBJECTIVES: To understand the similarities, differences, and relationships between three tools for performance and capacity assessment currently available for Maternal and Child Health (MCH) programs and for state and local health agencies. METHODS: Three tools for performance and capacity assessment currently available for Maternal and Child Health (MCH) programs and for state and local health agencies, the Title V MCH Block Grant Performance and Outcome Measures (Title V "24"), CAST-5, and the National Public Health Performance Standards Program (NPHPSP) were compared using two metrics, a conceptual model of the public health system, and a set of attributes related to the use of the instruments. RESULTS: Both CAST-5 and the NPHPSP are focused on the capacity and key processes (10 Essential Public Health Services) of the public health system, although CAST-5 is intended for capacity assessment and the NPHPSP is intended for performance measurement. The Title V "24" tool is also intended for performance measurement; however, its focus is on the outputs and outcomes of the health system. The Title V "24" tool is the only one of the three that is mandatory, and the only one whose results at the current time can be used to compare across entities. In addition, both the Title V "24" and the NPHPSP include explicit standards, while CAST-5 does not specify explicit standards against which to compare findings. CONCLUSIONS: While there are various tools available to MCH practitioners for capacity assessment and performance measurement, knowing how the tools relate to each other, and their defining characteristics, should lead to more effective and productive use. PMID- 12092981 TI - The practice of nurse-midwifery in the era of managed care: reports from the field. AB - OBJECTIVE: The purpose of this paper is to describe the reports of certified nurse-midwives (CNMs) about how changes in the financing and organization of health care in the late 1990s influenced their ability to serve vulnerable populations and provide a woman-centered, prevention-oriented midwifery model of care. METHODS: A 13-page survey was mailed to all CNMs ever certified by the American College of Nurse-Midwives (N = 6365) in July 1998. The survey included closed- and open-ended questions. A total of 2405 CNMs responded: of these, 2089 were in clinical practice during the study period (1997-98) and 82% of the 2089 (N = 1704) wrote responses to the open-ended questions and were included in the qualitative database. We present responses to the closed-ended questions about seven domains of practice and elaborate on three major themes identified through content analysis of the qualitative data. RESULTS: The majority (57%) reported that the changes in the larger health care environment had influenced their practices during 1997-98. The effects most frequently reported were 1) increased client loads (31%); 2) altered style of practice (30%): 3) inability to serve the same populations; (20%); 4) decreased client loads (20%); and 5) increased administrative duties (17%). Three major themes were identified and elaborated upon in the qualitative data: 1) challenges to the style of midwifery practice related to the managed care environment; 2) the loss of socially and economically at-risk women from CNMs' client base; and 3) barriers to high quality and comprehensive services for women. CONCLUSIONS: During the late 1990s as managed care was expanding and health systems were merging, a significant number of CNMs in the field described threats to their ability to sustain economically viable practices and a style of care consistent with the woman-centered, prevention oriented midwifery model. PMID- 12092982 TI - 2001 National MCH Epidemiology Awards: recognizing excellence. Maternal and Child Health. PMID- 12092983 TI - Preconceptional health promotion: opportunities abound. PMID- 12092984 TI - Preconception care: a systematic review. AB - OBJECTIVES: To perform a systematic review of published research trials of preconception care services to determine what evidence for effectiveness of care at improving the course of pregnancy or its outcomes has accumulated since the last major review in 1990. METHODS: The review was conducted adapting the systematic methods developed by the Cochrane Collaboration to collect evidence from published clinical research literature with as little bias as possible. The review included literature published after January 1990, and posted on MEDLINE by July 1999. RESULTS: Although more than 40 preconception risk conditions were searched and 470 articles were abstracted, only four problem areas and 19 research trials met the review criteria. New evidence of effectiveness was found for screening women who are seeking family planning for risk conditions; having sexually active women of reproductive age take dietary folate supplements; and providing women affected by certain metabolic conditions (diabetes and hyperphenylalanemia) with nutrition services. CONCLUSIONS: To help improve pregnancy outcomes MCH professionals need to promote the concept of readiness for pregnancy and help see that women are as healthy and appropriately nourished as possible before they become pregnant. PMID- 12092985 TI - Factors associated with smoking cessation among U.S. pregnant women. AB - OBJECTIVES: This study examines smoking and smoking cessation behaviors among U.S. pregnant women and seeks to identify the sociodemographic correlates of smoking cessation in pregnancy. METHODS: The 1998 NHIS Pregnancy and Smoking supplement was analyzed, including 5288 U.S. women (weighted to represent 13,714,358 women) who gave birth to a liveborn infant in the past 5 years. Four categories of smoking behavior were analyzed: nonsmoking at last pregnancy, persistent smoking throughout pregnancy, attempting unsuccessfully to quit during pregnancy, and successfully quitting during pregnancy. Logistic regression was used to isolate risk factors for each of the smoking behaviors and to examine factors associated with attempted and successful cessation. RESULTS: The women most likely to attempt to quit smoking in pregnancy were Hispanic women (OR = 3.09) and women who have smoked for less than 10 years (OR = 2.75 for women aged 18-24.) In general, for the groups at highest risk of smoking at the start of pregnancy, the odds of being a persistent smoker were higher than the odds of being an unsuccessful quitter, which in turn were higher than the odds of quitting successfully. The factors associated with attempts to quit included Hispanic ethnicity, higher education, above-poverty income, and shorter duration of smoking, while the combined effect of age and smoking duration was the only one significantly associated with successful quitting. In every age group, longer smoking duration was associated with lower likelihood of attempting to quit as well as successful quitting. CONCLUSIONS: The factors most strongly associated with attempts to quit smoking were Hispanic ethnicity and the combined effect of age and smoking duration. Future smoking cessation and relapse prevention programs should be developed, taking into consideration the critical factors of age, ethnicity, income, geography, and addiction. PMID- 12092986 TI - Differing postneonatal mortality rates of African-American and white infants in Chicago: an ecologic study. AB - OBJECTIVES: This study sought to determine whether neighborhood impoverishment explains the racial disparity in urban postneonatal mortality rates. METHODS: Stratified and multivariate logistic regression analyses were performed on the vital records of all African-Americans and whites born in Chicago by means of a linked 1992-1995 computerized birth-death file with appended 1990 U.S. census income and 1995 Chicago Department of Public Health data. Four community-level variables (low median family income, high rates of unemployment, homicide, and lead poisoning) were analyzed. Communities with one or more ecologic risk factors were classified as impoverished. RESULTS: The postneonatal mortality rate of African-Americans (N = 104,656) was 7.5/1000 compared to 2.7/1000 for whites (N = 52,954); relative risk (95% confidence interval) equaled 2.8 (2.3-3.3). Seventy nine percent of African-American infants compared to 9% of white infants resided in impoverished neighborhoods; p < 0.01. In impoverished neighborhoods, the adjusted odds ratio (controlling for infant and maternal individual-level risk factors) of postneonatal mortality for African-American infants equaled 1.5 (0.5 4.2). In nonimpoverished neighborhoods, the adjusted odds ratio of postneonatal mortality for African-American infants equaled 1.8 (1.1-2.9). CONCLUSIONS: We conclude that urban African-American infants who reside in nonimpoverished neighborhoods are at high risk for postneonatal mortality. PMID- 12092987 TI - Interdental mutans streptococci suppression in vivo: a comparison of different chlorhexidine regimens in relation to restorative material. AB - PURPOSE: To evaluate the suppressive effect of four different chlorhexidine (CHX) regimens on the interdental levels of mutans streptococci (MS) on enamel and various restorative materials in a group of adults. MATERIALS AND METHODS: 128 subjects with high salivary MS counts were allocated to (A) a single professional tooth cleaning with a 1% CHX gel, (B) duplicate applications of a dental varnish containing 1% CHX and 1% thymol, (C) a single application of a 40% CHX varnish, (D) daily tooth brushing with a 1% CHX gel for 2 weeks . Interdental plaque samples were collected at baseline, 1 week, and 3 months after termination of CHX treatment. RESULTS: The baseline MS levels were significantly higher in plaque from composite surfaces than in plaque from enamel surfaces. All methods decreased the interdental MS levels compared with the baseline on enamel, composite and amalgam surfaces. On both clinically sound enamel and amalgam surfaces, the outcomes in Groups B, C and D were similar and significantly better than in Group A. On composite surfaces, the 40% CHX varnish (C) and the CHX brushing (D) were significantly more effective in decreasing MS than the low-CHX varnish (B) and the professional CHX cleaning (A). PMID- 12092988 TI - Relation of enamel prism orientation to microtensile bond strength. AB - PURPOSE: To evaluate the effect of enamel prism orientation on microtensile bond strengths (MTBS) to enamel using two current bonding systems. MATERIALS AND METHODS: The crown segments of extracted human third molars were cut mesio distally to obtain buccal and lingual halves. One of the halves was cut horizontally at the middle of the crown to make a flat enamel surface parallel to prism orientation (parallel specimen). The other half was ground at the equator of the crown to produce a flat enamel surface perpendicular to the prism orientation (perpendicular specimen). Each surface was ground with 600 grit silicone carbide paper, treated with Clearfil Liner Bond IIsigma (LB) or Single Bond (SB) according to the manufacturers' instructions and resin composite, AP-X or Z100, respectively, was built up onto it. After storage in water at 37 degrees C for 24 hours, all bonded specimens were sectioned and trimmed for measuring MTBS. In MTBS, the parallel specimens were stressed perpendicular to the prism long axis and the perpendicular specimens were stressed parallel to the prism axis. RESULTS: MTBS of specimens stressed perpendicular to the prism long axis were 11.93 +/- 5.56 MPa for LB and 10.97 +/- 1.80 MPa for SB. MTBS of specimens stressed parallel to the prism axis in LB and SB were 19.07 +/- 6.35 MPa and 23.99 +/- 9.20 MPa, respectively. For each prism orientation, no difference in MTBS was found between the bonding systems (P> 0.05). Both systems showed lower MTBS in specimens stressed perpendicular to the prism long axis than in specimens stressed parallel to the prism axis. PMID- 12092989 TI - Effect of water quality on the bonding of resin to moist dentin. AB - PURPOSE: To test the effect of different water samples, used for rinsing the dentin, following acid conditioning, on the bond strength of a resin-based composite (RBC) restorative material (Z100), to dentin using a one-bottle dentin adhesive (Single Bond). MATERIALS AND METHODS: In preliminary experiments two experimental shear bond strength methods were compared, and one was selected which was deemed to permit more uniform application of the adhesive and better light curing. In the definitive experiments there were five experimental groups, according to rinsing water used (1) distilled water as control, (2) water with 3.5 ppm iodine, (3) water 3.0 ppm sodium hypochlorite, (4) water with 2% ethanol, (5) water contaminated with 6 x 10(4) cfu/mL of E. coli BH5V. For each experimental group the shear bond strength to moist dentin from 10 human intact impacted third molars was measured. The dentin samples were etched, washed and gently blotted to leave the dentin moist. Following application of the adhesive and resin, specimens were stored at 37 degrees C for 24 hours prior to testing in an Instron testing machine at a rate of 0.5 mm/minute. RESULTS: Data (in MPa +/- standard deviation) were: Group 1: 21.2 +/- 4.1; Group 2: 20.8 +/- 5.6; Group 3: 21.8 +/- 6.2; Group 4: 22.9 +/- 5.9; Group 5: 19.6 +/- 5.5. These data support the null hypothesis that the water types used in this work had no significant effect on the bonding of a resin composite to dentin (P= 0.74, ANOVA). PMID- 12092990 TI - Sealing ability of new generation adhesive-restorative materials placed on vital teeth. AB - The sealing ability of restorations is an important factor in order to prevent or reduce marginal gaps, which are often associated with postoperative sensitivity, caries and pulp disease. Usually the sealing ability of a new material is tested under laboratory conditions before being introduced on the market. However, in vitro evaluation of marginal integrity cannot predict in vivo performances of enamel-dentin bonding systems. A critical review of available data on sealing ability of last generation of adhesive-restorative materials placed on vital teeth is presented. The review includes the data of sealing performances of direct and indirect restorations, ethical considerations, influence of dental substrates on bonding, correlation between in vivo leakage studies and bond strength tests and microscopic evaluations. An in vivo protocol is presented in order to standardize leakage studies performed on vital teeth. A larger number of in vivo leakage tests are desirable in order to compare properties of new materials in multicenter studies. The ideal sequence of evaluation for new enamel dentin bonding systems should be: in vitro tests, short term in vivo evaluations and then clinical trials. PMID- 12092991 TI - The morphology and stability of resin-modified glass-ionomer adhesive at the dentin/resin-based composite interface. AB - PURPOSE: To evaluate the interface with tooth structure, mode of failure and stability of Fuji Bond LC (FBLC) resin-modified glass-ionomer bonding system. MATERIALS AND METHODS: The study was in three main parts. The first part with interfacial characteristics of FBLC. The micropermeability of the FBLC/dentin interface of restored cavities in extracted teeth, after fluorescent dye was introduced into the pulp chamber, was assessed by confocal microscopy. In other teeth, confocal microscopy was also used to examine trhe failure of FBLC bonded to flat dentin surfaces, on shear loading. In the second part, cervical cavities restored with FBLC (with and without light-curing) and a resin-based composite were examined for stability of the interface over the first 15 minutes. In the last part of the study, a new fluorescent dye was mixed with FBLC for restoration of cavities and subsequent examination using 2-photon imaging techniques. RESULTS: FBLC adapted well to tooth structure, had an interface with dentin that was permeable to fluid and displayed the absorption layer. Shear loading at the dentin interface resulted in complex failure with areas of cohesive failure where the absorption layer was present. Light-curing of FBLC seemed to produce a stable restorative while "co-curing" resulted in an unstable situation, with the appearance of an emmisable exudate from the FBLC. Advanced fluorescent imaging of the material itself seemed to suggest that there was a movement of aluminum ions in a manner similar to other glass-ionomer cements. The presence of HEMA in the matrix of the cement associated with the absorption layer was also demonstrated. PMID- 12092992 TI - Instrument sterilization and beyond. PMID- 12092993 TI - Firm enlargement of the dorsolateral tongue. PMID- 12092995 TI - Dental plaque removal with a novel battery-powered toothbrush. AB - PURPOSE: To compare the plaque removal efficacy of a positive control power toothbrush (Oral-B Ultra Plaque Remover) to an experimental power toothbrush (Crest SpinBrush) following a single use. MATERIALS AND METHODS: This study was a randomized, controlled, examiner-blind, 2-period crossover design which examined plaque removal with the two toothbrushes following a single use in 38 completed subjects. Plaque was scored before and after brushing using the Turesky Modification of the Quigley-Hein Index. RESULTS: Baseline plaque scores were 1.89 and 1.91 for the experimental toothbrush and control toothbrush treatment groups, respectively. With respect to all surfaces examined, the experimental toothbrush delivered an adjusted (via analysis of covariance) mean difference between baseline and post-brushing plaque scores of 0.46 while the control toothbrush delivered an adjusted mean difference of 0.45. These results were not statistically significant (P=0.645). A 95% one-sided upper confidence limit on the Ultra Plaque Remover minus SpinBrush difference in amount of plaque removed was calculated as 9.4% of the Ultra Plaque Remover adjusted mean. A common criterion for what is known as an "at least as good as" test is that the 95% one sided confidence limit on the product difference is below 10% of the control product mean. Using this criterion, the SpinBrush is at least as good as the Oral B Ultra Plaque Remover. With respect to buccal and lingual surfaces, the experimental toothbrush delivered very similar results relative to the control toothbrush. These results were also not statistically significant (P> 0.564). PMID- 12092994 TI - The effect of two power toothbrushes on calculus and stain formation. AB - PURPOSE: To compare the safety and efficacy with respect to the control of calculus and stain of two power toothbrushes. MATERIALS AND METHODS: This was a cross-over study involving a total of 81 subjects from a general population who used, in a randomized sequence, the Braun Oral-B 3D Excel (D17) and the Sonicare toothbrush with a conventional dentifrice, and a manual brush with a tartar control dentifrice. The manual toothbrush with tartar control dentifrice served as a positive control with respect to control of calculus. Following 9 weeks of manual brush use with a conventional, non-abrasive dentifrice, each test brush was used for a period of 9 weeks, after which subjects switched to the next brush in the sequence. Calculus was scored using the Volpe-Manhold Calculus Index and stain using the Lobene Stain Index. RESULTS: All three brushes were found to be safe as used in the study. All three toothbrushes significantly reduced the levels of calculus as compared to the control period. Reduction from baseline in the rate of calculus formation was greatest in the D17 group (63%), followed by the manual brush with tartar control dentifrice (60%) and Sonicare (44%). Both the D17 and the manual brush were significantly more effective than Sonicare (P< 0.001). The D17 was also more effective at controlling stain formation than either Sonicare or the manual brush with tartar control dentifrice at the gingival margin, the difference from Sonicare being statistically significant for all analyses (P< 0.0001). It is concluded that the D17 is significantly more effective in reducing both the rate of calculus and stain formation than the Sonicare toothbrush. PMID- 12092996 TI - Radiopacity of fiber-reinforced resin posts. AB - PURPOSES: (1) To determine the radiopacity of one titanium and seven fiber reinforced resin posts by densitometric analyses of radiographs, taken from the separate posts and from posts after seating in extracted root-treated human canine teeth with and without a radiopaque luting resin, respectively, and (2) To receive a ranking order by 20 dental practitioners of 16 randomly selected radiographs (2 for each post brand) by radiodensity and by acceptability assessment of the posts' radiographic appearance. MATERIALS AND METHODS: One reference titanium post (1) and the following 7 commercial fiber post brands were investigated: AEsthetiPlus/RTD (2), FiberKor/Jeneric Pentron (3), Light-Post/RTD (4), LightPost/Dentatus (5), Mirafit/Hager & Werken (6), Snowlight/Carbotech (7), and Snowpost/Carbotech (8). Standardized radiographs of the separate posts and of posts in extracted canines both before cementation and after cementation with Panavia 21 EX were produced and evaluated densitometrically together with a reference aluminum step-wedge. The cemented posts' radiodensities were determined from radiographs taken without and with 10 mm Plexiglass shielding. The optical densities were recorded in mm Al equivalents. Sixteen radiographs (2 for each post) were presented in random order to 20 dentists for ranking by decreasing radiopacity and by clinical acceptability scores from good (1), adequate (2), acceptable (3), not acceptable (4), to no radiopacity (5) of the posts' radiographic appearance. Statistical evaluation by ANOVA and Duncan post-hoc test at P< 0.05. RESULTS: Statistically the posts' radiodensities were significantly different. For separate and cemented posts, recorded from radiographs of Plexiglass shielded specimens, respectively, the following Duncan rankings were obtained: 1>7, 8>3>2, 4, 5>6 and 1>7, 8> 3, 5, 2, 6>6, 4. The dentist ranking by radiopacity scores was 1>7, 8>3>5,6>4>2, and by clinical acceptability scores 1>7, 8>3>2, 4, 5, 6. The titanium post (1) and posts 7 and 8 received scores better than acceptable, post 3 was assessed acceptable, whereas posts 2, 4, 5 and 6 were characterized as "not acceptable" or "not radiopaque". PMID- 12092998 TI - Properties of encapsulated and hand-mixed zinc phosphate dental cement. AB - PURPOSE: Dental cements can be supplied as loose powder and liquid or as encapsulated syringes. This study assessed the compressive strength of a recently marketed zinc phosphate encapsulated cement compared with a hand-mixed system according to the American Dental Association Specification. MATERIALS AND METHODS: The mean fracture strength, standard deviations and associated Weibull Moduli (m) of the encapsulated and hand-mixed cements were determined by compressive fracturing 30 cylindrical specimens (height 6.0 +/- 0.1 mm, diameter 4.0 +/- 0.1 mm). Scanning electron microscopy was employed to assess crack growth from indentations and cement morphology. Image analysis was used to investigate the influence of mixing regime on pore distribution within the cement samples. RESULTS: The compressive strength data showed variation in magnitude and reliability ranging from 42 +/- 8 MPa (m = 5.2 +/- 1.0) for the encapsulated cement filled directly from the capsule to 71 +/- 11 MPa (m = 6.3 +/- 1.3) for the hand-mixed cement. The encapsulated cement matrix was extensively porous, consisting of pores between 0.1-0.5 microm diameter, compared with the non-porous hand-mixed cement matrix. Larger pores (over 18 microm diameter) were related to air entrapment in the encapsulated cement on mixing while small pores (0.1-0.5 microm diameter) were indicative of vaporization porosity commonly seen with exothermic reactions. PMID- 12092997 TI - Effect of moist bonding on composite/enamel bond strength. AB - PURPOSE: To evaluate the effect of moist bonding on shear bond strength of resin based composite to enamel using different adhesive systems. MATERIALS AND METHODS: Six restorative systems were selected for this study: OptiBond FL/Prodigy, Solid Bond/Charisma F, Syntac Single Component/Tetric, Prime&Bond 2.1/Spectrum TPH, Single Bond/Z100, Etch&Prime 3.0/Degufill Mineral. Flat enamel surfaces were ground on the buccal and lingual aspects of 80 extracted human molars. OptiBond FL and Solid Bond were tested with and without primer application. Prior to application of the adhesives, the enamel was either carefully dried with compressed air (dry bonding) or blot dried with a cotton pellet (moist bonding). Shear bond strength was determined with a universal testing machine after 24-hour storage in 0.9% NaCl at 37 degrees C. RESULTS: Moist bonding did not significantly affect shear bond strength to enamel of the adhesives tested except for Solid Bond without primer application. Primer contamination of the etched enamel did not significantly influence bond strength, neither in the dry bonding nor in the moist bonding group. Of all adhesives tested in both groups, the highest mean bond strength was observed with Prime&Bond 2.1 and the lowest with Etch&Prime 3.0. PMID- 12092999 TI - Solvation of dried dentin matrix by water and other polar solvents. AB - PURPOSE: To develop a simple method for measuring the degree of solvation of dried, demineralized dentin matrix by water and other polar solvents. The null hypothesis was that there are no differences in expansion forces produced by different polar solvents. MATERIALS AND METHODS: Midcoronal dentin discs were prepared from extracted, unerupted human third molars. The discs were cut into square specimens with surface areas of 2 x 2, 3 x 3 and 4 x 4 mm and thicknesses of 0.5, 1.0 and 1.5 mm. After demineralization in 0.5 M EDTA (pH 7), the dimensions of the specimens were measured both wet and dry. Dry specimens were held between two parallel steel plates connected to a 50 N load cell which measured the solvation force when water or other polar solvents were added. After measuring the expansion force induced by water, the specimens were fixed in glutaraldehyde and the trials repeated. On additional specimens, repeated measures of expansion forces were obtained using water, methanol, ethanol, n propanol, n-butanol, ethylene glycol, formamide, hydroxyethylmethacrylate, N,N dimethyl formamide and acetone in unfixed specimens. RESULTS: Water produced hydration forces as high as 204 g before, and 428 g after glutaraldehyde treatment. The hydration force correlated better with specimen thickness than with surface area. Water solvated the matrix faster than methanol > ethanol > formamide > ethylene glycol. Hydroxyethylmethacrylate, N,N-dimethyl formamide and acetone were unable to solvate the dried matrix. Regression analysis of solvation force vs. Hansen's solubility parameters for dispersive, polar and hydrogen bonding forces demonstrated that solvation force correlations were highest with hydrogen bonding solubility parameters. Measurements of solvation forces provides a simple method for determining solvent-collagen matrix interactions. PMID- 12093000 TI - A novel peptide with ribonuclease and translation-inhibitory activities from fruiting bodies of the oyster mushroom Pleurotus ostreatus. AB - From the fresh fruiting bodies of the oyster mushroom a peptide with a molecular weight of 9 kDa and demonstrating a novel N-terminal sequence GPCYLVAFYESSGRR was isolated. The isolation procedure involved ion exchange chromatography on CM Sepharose and Mono S. The peptide was adsorbed on both types of chromatographic media. The peptide demonstrated a ribonuclease activity of 650 U/mg toward yeast transfer RNA. It inhibited cell-free translation in a rabbit reticulocyte lysate system with an IC50 of 15 nM. PMID- 12093001 TI - Peptoid residues and beta-turn formation. AB - A set of terminally protected tripeptoids containing a residue of either N methylglycine or N-isobutylglycine in position i + 1/i + 2 were synthesized and tested for intramolecularly H-bonded beta-turn formation. By exploiting FT-IR absorption and 1H NMR techniques, their folding tendencies were compared with those of a variety of reference peptides. The amount of beta-turn induction and the relative extent of the various types of intramolecularly H-bonded beta-turn conformers were determined in chloroform solution. PMID- 12093002 TI - Study of the conformational profile of the norbornane analogues of phenylalanine. AB - The conformational profile of the eight stereoisomeric 2-amino-3-phenylnorbornane 2-carboxylic acids (2-amino-3-phenylbicyclo[2.2.1]heptane-2-carboxylic acids) has been assessed by computational methods. These molecules constitute a series of four enantiomeric pairs that can be considered as rigid analogues of either L- or D-phenylalanine. The conformational space of their N-acetyl methylamide derivatives has been explored within the molecular mechanics framework, using the parm94 set of parameters of the AMBER force field. Local minimum energy conformations have been further investigated at the ab initio level by means of the Hartree-Fock and second order Moller-Plesset perturbation energy calculations using a 6-31G(d) basis set. The results of the present work suggest that the bulky norbornane structure induces two kinds of conformational constraints on the residues. On one hand, those of a steric nature directly imposed by the bicycle on the peptide backbone and, on the other hand, those that limit the orientations attainable by the phenyl ring which, in turn, reduces further the flexibility of the peptide backbone. A comparative analysis of the conformational profile of the phenylnorbornane amino acids with that of the norbornane amino acids devoid of the beta-phenyl substituent suggests that the norbornane system hampers the residue to adopt extended conformations in favour of C7-like structures. However, the bicycle itself does not impart a clear preference for any of the two possible C7 minima. It is the aromatic side chain, which is forced to adopt an almost eclipsed orientation, that breaks this symmetry introducing a marked preference for a single region of the (phi, psi) conformational space in each of the phenylalanine norbornane analogues investigated. PMID- 12093003 TI - Latent production of angiotensin I-converting enzyme inhibitors from buckwheat protein. AB - The latent production of angiotensin I-converting enzyme (ACE) Inhibitors from tartary buckwheat (BW) was investigated, and the peptides responsible for ACE inhibition characterized. Intact buckwheat was found to exhibit ACE inhibitory activity having an IC50 value of 3.0 mg/ml. The activity of the protein fraction (IC50: 0.36 mg protein/ml) was not enhanced by pepsin treatment. Pepsin, followed by chymotrypsin and trypsin hydrolysis, resulted in a significant increase in the ACE inhibitory activity (IC50: 0.14 mg protein/ml). The rutin contained in the buckwheat did not exhibit any ACE inhibition. A single oral administration of BW digest lowered the systolic blood pressure of a spontaneously hypertensive rat. Thus, BW proteins offer a potential resource for producing ACE inhibitory peptides during the digestion process. From the di-/tri-peptide fraction (DTPF) of the BW digest, inhibitory peptides were identified. The magnitude (%) of the total ACE inhibitory contribution of each identified peptide, relative to the overall inhibition of the DTPF, was about 41%. PMID- 12093004 TI - A comparison of three- and four-helix bundle TASP molecules. AB - We have designed, synthesized and characterized three- and four-helix bundle template-assembled synthetic proteins CTASPs). The TASPs were synthesized using disulphide bonds between the peptides and either the cyclotribenzylene (CTB) template, or the cavitand (BOWL) template, to form the three- and four helix bundles, respectively. The TASPs were constructed using peptides that were linked via their N-termini (peptide CGGGEELLKKXEELLKKG, where X = L, I, Nle or V), or via their C-termini (peptide GEELLKKLEELLKKGGGC). Each TASP was assayed for its structure, stability, 'native-like' characteristics and whether it was a monomer in solution. All TASPs were found to be highly helical, and highly resistant to chemical denaturation using guanidine hydrochloride (GnHCl). Analysis of the GnHCl-induced unfolding curves of the different TASPs demonstrated stability differences based on the number of helices in the bundle, the end of the helix that was attached to the template, and the identity of the core amino acid. The TASPs all had molten-globule structure, which is (generally) consistent with a degenerate sequence in the core. The four-helix bundle TASPs appeared to be monomers in solution, whereas there is some evidence that the three-helix bundle TASPs are weakly self-associating. PMID- 12093005 TI - CD4+ CD25+ suppressor T cells: more questions than answers. AB - Several mechanisms control discrimination between self and non-self, including the thymic deletion of autoreactive T cells and the induction of anergy in the periphery. In addition to these passive mechanisms, evidence has accumulated for the active suppression of autoreactivity by a population of regulatory or suppressor T cells that co-express CD4 and CD25 (the interleukin-2 receptor alpha chain). CD4+ CD25+ T cells are powerful inhibitors of T-cell activation both in vivo and in vitro. The enhancement of suppressor-cell function might prove useful for the treatment of immune-mediated diseases, whereas the downregulation of these cells might be beneficial for the enhancement of the immunogenicity of vaccines that are specific for tumour antigens. PMID- 12093006 TI - Cytotoxic T lymphocytes: all roads lead to death. AB - Cytotoxic T lymphocytes (CTLs) provide potent defences against virus infection and intracellular pathogens. However, CTLs have a dark side--their lytic machinery can be directed against self-tissues in autoimmune disorders, transplanted cells during graft rejection and host tissues to cause graft-versus host disease, which is one of the most serious diseases related to CTL function. Although this duplicitous behaviour might seem contradictory, both beneficial and detrimental effects are the result of the same effector proteins. So, an understanding of the mechanisms that are used by CTLs to destroy targets and a knowledge of pathogen immune-evasion strategies will provide vital information for the design of new therapies. PMID- 12093007 TI - Suppressors of cytokine signalling (SOCS) in the immune system. AB - The suppressors of cytokine signalling (SOCS) are a family of intracellular proteins, several of which have emerged as key physiological regulators of cytokine responses, including those that regulate the immune system. The SOCS proteins seem to regulate signal transduction by combining direct inhibitory interactions with cytokine receptors and signalling proteins with a generic mechanism of targeting associated proteins for degradation. Evidence is emerging for the involvement of SOCS proteins in diseases of the human immune system, which raises the possibility that therapeutic strategies that are based on the manipulation of SOCS activity might be of clinical benefit. PMID- 12093008 TI - No one is naive: the significance of heterologous T-cell immunity. AB - Memory T cells that are specific for one virus can become activated during infection with an unrelated heterologous virus, and might have roles in protective immunity and immunopathology. The course of each infection is influenced by the T-cell memory pool that has been laid down by a host's history of previous infections, and with each successive infection, T-cell memory to previously encountered agents is modified. Here, we discuss evidence from studies in mice and humans that shows the importance of this phenomenon in determining the outcome of infection. PMID- 12093009 TI - T-cell signalling and autoimmunity: molecular mechanisms of disease. AB - The genetic manipulation of mice has led to insights into the molecular mechanisms of autoimmune disease. Recent studies have begun to identify ways in which signalling cascades can be disrupted that preclude the development of autoimmunity. This review outlines a new model for the induction of T-cell mediated autoimmune diseases. I highlight recent data that illustrate the ways in which the altered survival of T cells and defects in the inhibitory signalling pathways of T cells can contribute to autoimmunity. PMID- 12093010 TI - Activation rules: the two-signal theories of immune activation. AB - Two-signal theories of lymphocyte activation have evolved considerably over the past 35 years. In this article, we examine the contemporary experimental observations and theoretical concerns that have helped to forge the most influential variants of the theory. We also propose that more-rigorous quantitative methods are required to sustain theoretical development in the future. PMID- 12093011 TI - The future of antigen-specific immunotherapy of allergy. AB - More than 25% of the population in industrialized countries suffers from immunoglobulin-E-mediated allergies. The antigen-specific immunotherapy that is in use at present involves the administration of allergen extracts to patients with the aim to cure allergic symptoms. However, the risk of therapy-induced side effects limits its broad application. Recent work indicates that the epitope complexity of natural allergen extracts can be recreated using recombinant allergens, and hypoallergenic derivatives of these can be engineered to increase treatment safety. It is proposed that these modified molecules will improve the current practice of specific immunotherapy and form a basis for prophylactic vaccination. PMID- 12093014 TI - International consensus statement on ADHD. January 2002. PMID- 12093013 TI - Critical issues in the prevention of violence-related behavior in youth. AB - Violence prevention programs with varying degrees of scientific support have proliferated in the United States and elsewhere. This paper previewed a broad range of programs involving youth, families, or systems that aimed to prevent or reduce violence-related behavior. The purpose of the review was to address critical issues concerning (1) target level of programming, (2) theory-driven versus problem-driven conceptualization, (3) cultural considerations, (4) developmental considerations, (5) intervention fidelity, and (6) outcome and impact assessment. Conclusions about these issues address tendencies and trends across programs. PMID- 12093012 TI - Familias Unidas: a family-centered ecodevelopmental intervention to reduce risk for problem behavior among Hispanic adolescents. AB - This paper describes the theoretical and empirical foundations of Familias Unidas, a multilevel, family-centered intervention designed to prevent problem behavior in Hispanic adolescents. The main theoretical tenets for the intervention model; an ecological-developmental perspective, the centrality of ethnic and cultural themes, application of empowerment principles, and a family focus are reviewed. The literature on the risk and protective factors that provided the justification for the intervention's targeted mediators and the core clinical applications that are intended to alter them are discussed. Familias Unidas engages Hispanic immigrant parents into an empowerment process in which they first build a strong parent-support network and then use the network to increase knowledge of culturally relevant parenting, strengthen parenting skills, and then apply these new skills in a series of activities designed to reduce risks frequently found in poor, urban environments. The available evidence supporting the efficacy of Familias Unidas is summarized, as are future goals and a current, second-generation application of the intervention. PMID- 12093015 TI - Comparative phylogenetic analysis of the evolution of semelparity and life history in salmonid fishes. AB - The selective pressures involved in the evolution of semelparity and its associated life-history traits are largely unknown. We used species-level analyses, independent contrasts, and reconstruction of ancestral states to study the evolution of body length, fecundity, egg weight, gonadosomatic index, and parity (semelparity vs. degree of iteroparity) in females of 12 species of salmonid fishes. According to both species-level analysis and independent contrasts analysis, body length was positively correlated with fecundity, egg weight, and gonadosomatic index, and semelparous species exhibited a significantly steeper slope for the regression of egg weight on body length than did iteroparous species. Percent repeat breeding (degree of iteroparity) was negatively correlated with gonadosomatic index using independent contrasts analysis. Semelparous species had significantly larger eggs by species-level analysis, and the egg weight contrast for the branch on which semelparity was inferred to have originated was significantly larger than the other egg weight contrasts, corresponding to a remarkable increase in egg weight. Reconstruction of ancestral states showed that egg weight and body length apparently increased with the origin of semelparity, but fecundity and gonadosomatic index remained more or less constant or decreased. Thus, the strong evolutionary linkages between body size, fecundity, and gonadosomatic index were broken during the transition from iteroparity to semelparity. These findings suggest that long distance migrations, which increase adult mortality between breeding episodes, may have been necessary for the origin of semelparity in Pacific salmon, but that increased egg weight, leading to increased juvenile survivorship, was crucial in driving the transition. Our analyses support the life-history hypotheses that a lower degree of repeat breeding is linked to higher reproductive investment per breeding episode, and that semelparity evolves under a combination of relatively high juvenile survivorship and relatively low adult survivorship. PMID- 12093016 TI - Evidence for historical introgression along a contact zone between two species of char (Pisces: Salmonidae) in northwestern North America. AB - Phylogeographic analyses can yield valuable insights into the geographic and historical contexts of contact and hybridization between taxa. Two species of char (Salmonidae), Dolly Varden (Salvelinus malma) and bull trout (S. confluentus) have largely parapatric distributions in watersheds of northwestern North America. They are, however, sympatric in several localities and hybridization and some introgression occurs across a broad area of contact. We conducted a comparative phylogenetic analysis of Dolly Varden and bull trout to gain a historical perspective of hybridization between these species and to test for footprints of historical introgression. We resolved two major Dolly Varden mitochondrial DNA (mtDNA) clades (with 1.4-2.2% sequence divergence between haplotypes) that had different geographical distributions. Clade N is distributed across most of the range of Dolly Varden, from southern British Columbia through to the Kuril Islands in Asia. Clade S had a much more limited distribution, from Washington state, at the southern limit of the Dolly Varden range, to the middle of Vancouver Island. The distribution and inferred ages of the mtDNA clades suggested that Dolly Varden survived the Wisconsinan glaciation in a previously unsuspected refuge south of the ice sheet, and that Dolly Varden and bull trout were probably in continuous contact over most of the last 100,000 years. When bull trout were included in the phylogenetic analysis, however, the mtDNA of neither species was monophyletic: Clade S Dolly Varden clustered within the bull trout mtDNA clade. This pattern was discordant with two nuclear phylogenies produced (growth hormone 2 and rRNA internal transcribed sequence 1), in which Dolly Varden and bull trout were reciprocally monophyletic. This discordance between mtDNA- and nDNA-based phylogenies indicates that historical introgression of bull trout mtDNA into Dolly Varden occurred. Percent sequence divergence within these introgressed Dolly Varden (clade S) was 0.2-0.6%, implying that the introgression occurred prior to the most recent glaciation. Our analysis and other evidence of contact between divergent lineages in northwestern North America strongly suggests that the area may be the site of previously unsuspected suture zones of aquatic biotas. PMID- 12093017 TI - Deleterious alleles and differential viability in progeny of natural hemiclonal frogs. AB - Abstract.-Spontaneous deleterious mutations are expected to accumulate through Muller's ratchet in clonally reproducing organisms and may lead to their extinction. We study deleterious mutations and their effects in a system of European frogs. Rana esculenta (RL), natural hybrids R. ridibunda (RR) X R. lessonae (LL), reproduce hemiclonally; both sexes exclude the L genome in the germ line and produce unrecombined R gametes; hybridity is restored each generation by matings of RL with coexisting LL. Different allozyme-defined hybrid hemiclones (R genome haplotypes) are thought to have originated independently from primary hybridizations RR x LL. Natural matings between two hybrids usually lead to inviable RR tadpoles. This inviability is thought to result from unmasked deleterious alleles on the clonally transmitted R genomes. Most simply it reflects homozygosity for recessive deleterious alleles at particular loci; alternatively (consistent with absence of RR adults in multiclonal populations) it may reflect hemiclone-specific sets of incompletely recessive deleterious mutations that cumulatively cause inviability when two such genomes are combined. If inviability results from the former, progeny of two hybrids of different hemiclones, whether allopatric or coexisting, should be viable, because it is improbable that their R genomes share recessive deleterious alleles at the same set of loci; if inviability results from the latter, progeny of hybrids of different hemiclones should be inviable, especially when hybrid lineages are old. We tested these hypotheses in artificial crosses, using frogs from three regions: hemiclonal hybrids outside R. ridibunda's range from northern Switzerland (two abundant coexisting allozyme-defined hemiclones; estimated lineage age < or = 5,000 generations) and from Sicily, Italy (one hemiclone; estimated age > or = 25,000 generations) and R. ridibunda from Poland. We generated RR progeny, which we reared under benign conditions in the laboratory, by crossing (1) two hybrids from the same region (H x H local); (2) two hybrids from different regions (H X H foreign); (3) hybrids and R. ridibunda (H X R); and (4) two R. ridibunda (R X R). Survival to metamorphosis was similar and high for R x R, H X H foreign, and H X R, whereas all tadpoles of H X H local died before metamorphosis. This supports the hypothesis that homozygosity for recessive deleterious mutations at particular loci causes inviability. Crosses within and between the two coexisting hemiclones from Switzerland were, however, equally inviable. This result may reflect episodic sexual recombination in RR progeny from exceptional successful interclonal hybrid X hybrid matings, followed by matings of such RR with LL. This process would both slow down or halt Muller's ratchet and disrupt genetic independence of coexisting hemiclones, so that the same remaining deleterious R alleles could exist in different allozyme-defined hemiclones. Whereas all data are consistent with the prediction of Muller's ratchet operating on clonally transmitted R genomes of natural hybrid lineages, they are insufficient to demonstrate such operation, because deleterious recessives that mutated after clone formation and those that preexisted in the R. ridibunda source populations that formed the hemiclonal lineages are not distinguished. The possibility of episodic sexual recombination must be carefully taken into account when studying Muller's ratchet in natural populations of this Rana system. PMID- 12093018 TI - Dwarfism in insular sloths: biogeography, selection, and evolutionary rate. AB - The islands of Bocas del Toro, Panama, were sequentially separated from the adjacent mainland by rising sea levels during the past 10,000 years. Three-toed sloths (Bradypus) from five islands are smaller than their mainland counterparts, and the insular populations themselves vary in mean body size. We first examine relationships between body size and physical characteristics of the islands, testing hypotheses regarding optimal body size, evolutionary equilibria, and the presence of dispersal in this system. To do so, we conduct linear regressions of body size onto island area, distance from the mainland, and island age. Second, we retroactively calculate two measures of the evolutionary rate of change in body size (haldanes and darwins) and the standardized linear selection differential, or selection intensity (i). We also test the observed morphological changes against models of evolution by genetic drift. The results indicate that mean body size decreases linearly with island age, explaining up to 97% of the variation among population means. Neither island area nor distance from the mainland is significant in multiple regressions that include island age. Thus, we find no evidence for differential optimal body size among islands, or for dispersal in the system. In contrast, the dependence of body size on island age suggests uniform directional selection for small body size in the insular populations. Although genetic drift cannot be discounted as the cause for this evolution in body size, the probability is small given the consistent direction of evolution (repeated dwarfism). The insular sloths show a sustained rate of evolution similar to those measured in haldanes over tens of generations, appearing to unite micro- and macroevolutionary time scales. Furthermore, the magnitude and rate of this example of rapid differentiation fall within predictions of theoretical models from population genetics. However, the linearity of the relationship between body size and island age is not predicted, suggesting that either more factors are involved than those considered here, or that theoretical advances are necessary to explain constant evolutionary rates over long time spans in new selective environments. PMID- 12093019 TI - The cost of fluctuating inbreeding depression. AB - We present a phenotypic model for the evolution of self-fertilization in an infinite population of annual hermaphrodites for the case in which fitness and inbreeding depression vary among generations (e.g., due to fluctuations in the environment from year to year). Conditions for the evolution of selfing, mixed mating, and outcrossing are derived and are compared with results from numerical calculations that assume a normal distribution of inbreeding depression. In contrast to the situation in which inbreeding depression does not vary, when inbreeding depression fluctuates in a stochastic manner among generations with a mean less than 0.5, selfing is not necessarily selected. Thus, fluctuating inbreeding depression can be viewed as an additional cost of selfing that may stabilize mixed mating systems. These results emphasize the need to take into account fluctuating inbreeding depression in empirical studies aimed at understanding mating system evolution in annuals. PMID- 12093020 TI - Environmental dependency in the expression of costs of tolerance to deer herbivory. AB - Plant tolerance to natural enemy damage is a defense strategy that minimizes the effects of damage on fitness. Despite the apparent benefits of tolerance, many populations exhibit intermediate levels of tolerance, indicating that constraints on the evolution of tolerance are likely. In a field experiment with the ivyleaf morning glory, costs of tolerance to deer herbivory in the form of negative genetic correlations between deer tolerance and fitness in the absence of damage were detected. However, these costs were detected only in the presence of insect herbivores. Such environmental dependency in the expression of costs of tolerance may facilitate the maintenance of tolerance at intermediate levels. PMID- 12093021 TI - Geographic variation for wing shape in Drosophila serrata. AB - Geographic variation in wing shape in female Drosophila serrata was examined by characterizing isofemale strains from 19 localities collected along a transect on the eastern coast of Australia. Shape variation was analyzed by Procrustes superimposition of landmark data followed by canonical variate analysis. The first extracted canonical variate showed a nonlinear association with latitude and accounted for 43% of the variance. There was a sharp increase in this variate at low latitudes as well as a gradual increase at high latitudes. These shape changes were associated with two landmarks at the edge of the wing. There was also a linear change in wing aspect. The isofemale heritability for two measures of shape was around 30%. Allometric relationships were weak both between localities and among isofemale strains within localities. The possibility that wing shape parameters are under selection independent of wing size is discussed. PMID- 12093022 TI - A genetic correlation between age at pupation and melanization immune response of the yellow fever mosquito Aedes aegypti. AB - To investigate the evolutionary cost of an immune response, we selected six lines of the mosquito Aedes aegypti for earlier or later pupation and measured the extent to which this selection procedure changed the mosquito's ability to encapsulate and melanize a negatively charged Sephadex bead. After 10 generations of selection, the age at pupation in the two selection regimes differed by about 0.7 days, accompanied by an increase of wing length of the mosquitoes selected for late pupation. Among the mosquitoes that had been selected for early pupation, only 6% had strongly or completely melanized the bead, while among the individuals that had been selected for late pupation, 32% had melanized the bead. Thus, our results suggest a genetic correlation between age at pupation and immunocompetence. As a consequence, mosquitoes that respond to increased intense parasite pressure with more effective immunity are predicted to pay for the increased defense with slower development. PMID- 12093023 TI - The effect of epistasis on the excess of the additive and nonadditive variances after population bottlenecks. AB - The effect of population bottlenecks on the components of the genetic variance generated by two neutral independent epistatic loci has been studied theoretically (VA, additive; VD, dominant; VAA, additive x additive; VAD, additive x dominant; VDD; dominant x dominant components of variance). Nonoverdominance and overdominance models were considered, covering all possible types of marginal gene action at the single locus level. The variance components in an infinitely large panmictic population (ancestral components) were compared with their expected values at equilibrium, after t consecutive bottlenecks of equal size N (derived components). Formulae were obtained in terms of allele frequencies and effects at each locus and the corresponding epistatic value. An excess of VA after bottlenecks can be assigned to two sources: (1) the spatiotemporal changes in the marginal average effects of gene substitution alpha(i), which are equal to zero only for additive gene action within and between loci; and (2) the covariance between alpha2(i) and the heterozygosity at the loci involved, which is generated by dominance, with or without epistasis. Numerical examples were analyzed, indicating that an increase in VA after bottlenecks will only occur if its ancestral value is minimal or very small. For the nonoverdominance model with weak reinforcing epistasis, that increase has been detected only for extreme frequencies of the negative allele at one or both loci. With strong epistasis, however, this result can be extended to a broad range of intermediate frequencies. With no epistasis, the same qualitative results were found, indicating that dominance can be considered as the primary cause of an increase in VA following bottlenecks. In parallel, the derived total nonadditive variance exceeded its ancestral value (V(NA) = V(D) + V(AA) + V(AD) + V(DD)) for a range of combinations of allele frequencies covering those for an excess of VA and for very large frequencies of the negative allele at both loci. For the overdominance model, an increase in V(A) and V(NA) was respectively observed for equilibrium (intermediate) frequencies at one or both loci or for extreme frequencies at both loci. For all models, the magnitude of the change of V(A) and V(NA) was inversely related to N and t. At low levels of inbreeding, the between-line variance was not affected by the type of gene action. For the models considered, the results indicate that it is unlikely that the rate of evolution may be accelerated after population bottlenecks, in spite of occasional increments of the derived V(A) over its ancestral value. PMID- 12093024 TI - The role of size-specific predation in the evolution and diversification of prey life histories. AB - Some of the best empirical examples of life-history evolution involve responses to predation. Nevertheless, most life-history theory dealing with responses to predation has not been formulated within an explicit dynamic food-web context. In particular, most previous theory does not explicitly consider the coupled population dynamics of the focal species and its predators and resources. Here we present a model of life-history evolution that explores the evolutionary consequences of size-specific predation on small individuals when there is a trade-off between growth and reproduction. The model explicitly describes the population dynamics of a predator, the prey of interest, and its resource. The selective forces that cause life-history evolution in the prey species emerge from the ecological interactions embodied by this model and can involve important elements of frequency dependence. Our results demonstrate that the strength of the coupling between predator and prey in the community determines many aspects of life-history evolution. If the coupling is weak (as is implicitly assumed in many previous models), differences in resource productivity have no effect on the nature of life-history evolution. A single life-history strategy is favored that minimizes the equilibrium resource density (if possible). If the coupling is strong, then higher resource productivities select for faster growth into the predation size refuge. Moreover, under strong coupling it is also possible for natural selection to favor an evolutionary diversification of life histories, possibly resulting in two coexisting species with divergent life-history strategies. PMID- 12093025 TI - A phylogenetic study of pollinator conservatism among sexually deceptive orchids. AB - Orchids of the genus Chiloglottis are pollinated through the sexual deception of male wasps mainly from the genus Neozeleboria (Tiphiidae: Thynninae). The orchids mimic both the appearance and sex pheromones of wingless female thynnines but provide no reward to the deceived males. Despite the asymmetry of this interaction, strong pollinator specificity is typical. Such plant-pollinator interactions would seem to be relatively flexible in the plant's adaptive response to variation in the local pollinator resource. However, we present DNA sequence data on both orchids and wasps that demonstrate a pattern of pollinator conservatism operating at a range of taxonomic levels. Sequence data from the wasps indicate 15 of 16 Chiloglottis pollinators are closely related members of one clade of Thynninae. A pattern of congruence between orchid and wasp phylogenies is also demonstrated below the generic level, such that related orchids tend to use related thynnine wasps as specific pollinators. Comparative physiological data on the wasp responses to the floral scents of two Chiloglottis species and one outgroup, Arthrochilus, indicate similar attractive volatile chemicals are used by related orchid taxa. By extension, we infer a similarity of sex pheromone signals among related thynnines. Thus, the conservative pattern of pollinator change in sexually deceptive orchids may reflect phylogenetic patterns in the sex pheromones of their pollinators. PMID- 12093026 TI - Resistance and tolerance in a host plant-holoparasitic plant interaction: genetic variation and costs. AB - Host organisms are believed to evolve defense mechanisms (i.e., resistance and/or tolerance) under selective pressures exerted by natural enemies. A prerequisite for the evolution of resistance and tolerance is the existence of genetic variation in these traits for natural selection to act. However, selection for resistance and/or tolerance may be constrained by negative genetic correlations with other traits that affect host fitness. We studied genetic variation in resistance and tolerance against parasitic infection and the potential fitness costs associated with these traits using a novel study system, namely the interaction between a flowering plant and a parasitic plant. In this system, parasitic infection has significant negative effects on host growth and reproduction and may thus act as a selective agent. We conducted a greenhouse experiment in which we grew host plants, Urtica dioica, that originated from a single natural population and represented 20 maternal families either uninfected or infected with the holoparasitic dodder, Cuscuta europaea. that originated from the same site. We calculated correlations among resistance, tolerance, and host performance to test for costs of resistance and tolerance. We measured resistance as parasite performance (quantitative resistance) and tolerance as the slopes of regressions relating the vegetative and reproductive biomass of host plants to damage level (measured as parasite biomass). We observed significant differences among host families in parasite resistance and in parasite tolerance in terms of reproductive biomass, a result that suggests genetic variation in these traits. Furthermore, we found differences in resistance and tolerance between female and male host plants. In addition, the correlations indicate costs of resistance in terms of host growth and reproduction and costs of tolerance in terms of host reproduction. Our results thus indicate that host tolerance and resistance can evolve as a response to infection by a parasitic plant and that costs of resistance and tolerance may be one factor maintaining genetic variation in these traits. PMID- 12093027 TI - Accelerated molecular evolution in halophilic crustaceans. AB - In contrast to the stable ionic composition of the oceans, inland waters show striking diversity, possessing salt concentrations varying from I mM to 5 M. Although species diversity is highest in fresh water, some lineages have colonized hypersaline environments where they encounter elevated levels of both ultraviolet (UV) radiation and osmotic stress. This study compares rates of evolution in halophilic and freshwater taxa for two groups of microcrustaceans, anostracans and daphniids, from Australia and North America. The results establish that halophilic species show consistent rate acceleration, involving elevated levels of both insertion/deletion events and of nucleotide substitutions. The elevated pace of molecular evolution does not appear to be linked to selection or to other agents that are known to influence the supply rate of mutations, such as UV exposure, generation length, or shifts in metabolic rate. However, variance in ionic strength, which is known to have potent effects on DNA-protein interactions as well as on the structural properties of DNA and proteins, might account for the lowered fidelity of DNA replication in life from hypersaline settings. Regardless of its cause, the consistent rate acceleration in halophiles suggests that past efforts to employ sequence divergences to date events, such as the age of asexual lineages in Artemia, have resulted in serious overestimates. More generally, the results indicate that coordinated shifts in rates of molecular evolution may occur in lineages exposed to extreme environmental conditions. PMID- 12093028 TI - The role of parental age effects on the evolution of aging. AB - Many studies have found that older parents have shorter-lived offspring. However, the evolutionary significance of these findings is poorly understood. We carried out large-scale demographic experiments to examine the direct effect of maternal age and paternal age on offspring aging in inbred and outbred strains of the fruit fly Drosophila melanogaster. We found that the age of mothers and, to a lesser extent, the age of fathers can have a large influence on both offspring longevity and the shape of the age-specific mortality trajectory. In two independent experiments we found that older mothers generally produced shorter lived offspring, although the exact effect of maternal age on offspring longevity differed among strains. These results suggest that maternal age effects on progeny aging may influence the evolution of aging. PMID- 12093029 TI - Correlated evolution of male and female morphologles in water striders. AB - Sexually antagonistic coevolution may be an important force in the evolution of sexual dimorphism. We undertake a comparative study of correlated evolution of male and female morphologies in a clade of 15 water strider species in the genus Gerris (Heteroptera: Gerridae). Earlier studies have shown that superfluous matings impose costs on females, including increased energetic expenditure and predation risk, and females therefore resist males with premating struggles. Males of some species possess grasping structures and females of some species exhibit distinct antigrasping structures, which are used to further the interests of each sex during these premating struggles. We use this understanding, combined with coevolutionary theory, to derive a series of a priori predictions concerning both the types of traits in the two sexes that are expected to coevolve and the coevolutionary dynamics of these traits expected under sexually antagonistic coevolution. We then assess the actual pattern of correlated evolution in this clade with new morphometric methods combined with standard comparative techniques. The results were in agreement with the a priori predictions. The level of armament (different abdominal structures in the two sexes) was closely correlated between the sexes across species. Males are well adapted to grasping females in species in which females are well adapted to thwart harassing males and vice versa. Furthermore, our comparative analyses supports the prediction that correlated evolution of armament in the two sexes should be both rapid and bidirectional. PMID- 12093030 TI - Evolution after the flood: phylogeography of the desert fish Utah Chub. AB - The Bonneville Basin and upper Snake River drainage of western North America underwent extensive hydrological changes during the late Pleistocene, potentially influencing the geographic distribution and evolutionary trajectories of aquatic species that occupied this region. To test this hypothesis, I reconstructed the phylogeographic history of the desert fish Utah chub (Gila atraria) by examining 16 populations that span the natural distribution of this species across the Bonneville Basin and upper Snake River. I compared mitochondrial control region sequences (934 bp) among 77 individuals revealing 24 unique haplotypes. Geographic and phylogenetic relationships among haplotypes were explored using parsimony, maximum likelihood, nested clade analysis, and analysis of molecular variance. I found that G. atraria is composed of two distinct clades that represent an early Pleistocene split between the upper Snake River and Bonneville Basin. Within each of these clades, geographic structuring was highly concordant with the hydrological history of late Pleistocene Lake Bonneville and the upper Snake River, suggesting that glacial-induced shifts in climate and unpredictable geological events have played a major role in shaping genetic subdivision among populations. To examine the effects of vicariant events on phenotypic divergence among Utah chub populations, I mapped chub life histories to the control region haplotype network. I found a nonrandom association between haplotypes and life history phenotypes. These results suggest that historical events responsible for population fragmentation may have also contributed to phenotypic shifts in life histories, both indirectly by limiting gene flow among populations and directly by altering the selective environments where populations persisted. PMID- 12093031 TI - Evolutionary history of the parrotfishes: biogeography, ecomorphology, and comparative diversity. AB - The family Scaridae comprises about 90 species of herbivorous coral reef, rock reef, and seagrass fishes. Parrotfishes are important agents of marine bioerosion who rework the substrate with their beaklike oral jaws. Many scarid populations are characterized by complex social systems including highly differentiated sexual stages, territoriality, and the defense of harems. Here, we test a hypothesis of relationships among parrotfish genera derived from nearly 2 kb of nuclear and mitochondrial DNA sequence. The DNA tree is different than a phylogeny based on comparative morphology and leads to important reinterpretations of scarid evolution. The molecular data suggest a split among seagrass and coral reef associated genera with nearly 80% of all species in the coral reef clade. Our phylogenetic results imply an East Tethyan origin of the family and the recurrent evolution of excavating and scraping feeding modes. It is likely that ecomorphological differences played a significant role in the initial divergence of major scarid lineages, but that variation in color and breeding behavior has triggered subsequent diversification. We present a two phase model of parrotfish evolution to explain patterns of comparative diversity. Finally, we discuss the application of this model to other adaptively radiating clades. PMID- 12093032 TI - Historical biogeography of the livebearing fish genus Poeciliopsis (Poeciliidae: Cyprinodontiformes). AB - To assess the historical biogeography of freshwater topminnows in the genus Poeciliopsis, we examined sequence variation in two mitochondrial genes, cytochrome b (1140 bp) and NADH subunit 2 (1047 bp). This widespread fish genus is distributed from Arizona to western Colombia, and nearly half of its 21 named species have distributions that border on the geologically active Trans-Mexican Volcanic Belt (TMVB), a region that defines the uplifted plateau (Mesa Central) of Mexico. We used the parametric bootstrap method to test the hypothesis that a single vicariant event associated with the TMVB was responsible for divergence of taxa found to the north and south of this boundary. Because the single-event hypothesis was rejected as highly unlikely, we hypothesize that at least two geological events were responsible for divergence of these species. The first (8 16 million years ago) separated ancestral populations that were distributed across the present TMVB region. A second event (2.8-6.4 million years ago) was associated with northward dispersal and subsequent vicariance of two independent southern lineages across the TMVB. The geological history of this tectonically and volcanically active region is discussed and systematic implications for the genus are outlined. PMID- 12093033 TI - Ancient versus reticulate origin of a hemiclonal lineage. AB - Crossing experiments revealed that a diploid hybridogenetic fish (genus Poeciliopsis) from the Rio Mocorito (Sinaloa, Mexico) is trihybrid. Its haploid maternal genome is inherited clonally (i.e., hemiclonally), and it expresses a mixture of morphological traits found in the closely related species P. monacha and P. viriosa. Its haploid paternal genome is replaced in each generation by mating with males of a more distantly related sexual species, P. lucida. However, expression of mixed (monacha X viriosa) traits by this hemiclone is also consistent with retention of shared ancestral polymorphisms. If true, this hemiclonal lineage would be one of the few examples of an ancient asexual taxon. We used mitochondrial DNA and allozymes to test whether the maternal progenitor of the Mocorito hybridogen was a recent P. monacha X P. viriosa hybrid or a remnant of their most recent common ancestor. Our results clearly link the hemiclonal genome to contemporary P. monacha and therefore support the hypothesis of a recent origin. Additionally, our findings suggest that this unisexual fish may serve as a vehicle for introgression between two allopatric sexual species. PMID- 12093034 TI - Evolution of behavior and neural control of the fast-start escape response. AB - The fast-start startle behavior is the primary mechanism of rapid escape in fishes and is a model system for examining neural circuit design and musculoskeletal function. To develop a dataset for evolutionary analysis of the startle response, the kinematics and muscle activity patterns of the fast-start were analyzed for four fish species at key branches in the phylogeny of vertebrates. Three of these species (Polypterus palmas, Lepisosteus osseus, and Amia calva) represent the base of the actinopterygian radiation. A fourth species (Oncorhynchus mykiss) provided data for a species in the central region of the teleost phylogeny. Using these data, we explored the evolution of this behavior within the phylogeny of vertebrates. To test the hypothesis that startle features are evolutionarily conservative, the variability of motor patterns and kinematics in fast-starts was described. Results show that the evolution of the startle behavior in fishes, and more broadly among vertebrates, is not conservative. The fast-start has undergone substantial change in suites of kinematics and electromyogram features, including the presence of either a one- or a two-stage kinematic response and change in the extent of bilateral muscle activity. Comparative methods were used to test the evolutionary hypothesis that changes in motor control are correlated with key differences in the kinematics and behavior of the fast-start. Significant evolutionary correlations were found between several motor pattern and behavioral characters. These results suggest that the startle neural circuit itself is not conservative. By tracing the evolution of motor pattern and kinematics on a phylogeny, it is shown that major changes in the neural circuit of the startle behavior occur at several levels in the phylogeny of vertebrates. PMID- 12093035 TI - White spot syndrome virus (WSSV) PCR-positive Artemia cysts yield PCR-negative nauplii that fail to transmit WSSV when fed to shrimp postlarvae. AB - Positive results were obtained with nested white spot syndrome virus (WSSV) diagnostic PCR performed on 5 commercial brands of dry-packed Artemia cysts using several WSSV genomic sequence-specific primers. In 2 brands, PCR and nucleotide sequence analysis found C-->T and C-->G point mutations in the pms 146 WSSV amplicon, but in all 5 brands, the nucleotide sequences that were successfully amplified by the rrl, rr2 and tk-tmk gene-specific primer sets were identical to those of Penaeus monodon WSSV. However, despite the inarguable presence of WSSV or WSSV-like template DNA, we were unable to detect WSSV by PCR in hatched nauplii derived from PCR-positive cysts or in P. monodon postlarvae fed Artemia nauplii hatched from such cysts. Most simply, these results suggested that the cysts were externally contaminated with WSSV or WSSV-like template material that was removed during hatching and washing of the nauplii. Given the small sequence variations found, it may also have been a variety of WSSV non-infectious for P. monodon or Artemia and derived from other crustaceans or arthropods in the Artemia environment. However, we could not establish this conclusively and a small possibility remained that the PCR template in these tests was derived from WSSV template present internally in the cysts and derived from infected Artemia adults. However small, this possibility must be vigorously tested, given the impact that a positive outcome could have on the shrimp industry. PMID- 12093036 TI - Detection of white spot syndrome virus (WSSV) of shrimp by means of monoclonal antibodies (MAbs) specific to an envelope protein (28 kDa). AB - The vp28 gene encoding an envelope protein (28 kDa) of white spot syndrome virus (WSSV) was amplified from WSSV-infected tiger shrimp that originated from Malaysia. Recombinant VP28 protein (r-28) was expressed in Escherichia coli and used as an antigen for preparation of monoclonal antibodies (MAbs). Three murine MAbs (6F6, 6H4 and 9C10) that were screened by r-28 antigen-based enzyme-linked immunosorbent assay (ELISA) were also able to recognize viral VP28 protein as well as r-28 on Western blot. Three non-overlapping epitopes of VP28 protein were determined using the MAbs in competitive ELISA; thus, an antigen-capture ELISA (Ac-ELISA) was developed by virtue of these MAbs. Ac-ELISA can differentiate WSSV infected shrimp from uninfected shrimp and was further confirmed by a polymerase chain reaction (PCR) and Western blot. Approximately 400 pg of purified WSSV sample and 20 pg of r-28 could be detected by Ac-ELISA, which is comparable in sensitivity to PCR assay but more sensitive than Western blot in the detection of purified virus. Hemolymph and tissue homogenate samples collected from a shrimp farm in Malaysia during December 2000 and July 2001 were also detected by Ac ELISA and PCR with corroborating results. PMID- 12093037 TI - A syntenin-like protein with postsynaptic density protein (PDZ) domains produced by black tiger shrimp Penaeus monodon in response to white spot syndrome virus infection. AB - We report the isolation and characterization of products from a subtractive cDNA library from the haemolymph of Penaeus monodon experimentally infected with white spot syndrome virus (WSSV). One cDNA derived from up-regulated mRNA was identified. A homology search indicated similarity to the putative protein syntenin (TE8). The nearly complete nucleotide sequence of TE8 was obtained by rapid amplification of cDNA (RACE). Its putative protein product contained a tandem repeat of PDZ domains (postsynaptic density protein or PSD-95, DlgA and ZO 1). We propose that TE8 may function as an adapter that couples PDZ-binding protein(s) in a signaling pathway involved in the shrimp response to WSSV. PMID- 12093039 TI - Detection of Piscirickettsia salmonis in fish tissues by an enzyme-linked immunosorbent assay using specific monoclonal antibodies. AB - An enzyme-linked immunosorbent assay (ELISA) for the detection of Piscirickettsia salmonis in fish tissue samples was developed. The test uses a combination of different monoclonal antibodies specific against P. salmonis in the capture step of the assay. The antibodies 7G4, 6E2 and 2C1 chosen for the capture step are bound to the solid support with an adhesive protein purified from a bivalve mollusc, resulting in a high yield of adsorption and binding stability. The monoclonal antibody 7G4, used as a second antibody, is conjugated to horseradish peroxidase. The resulting ELISA test detected 7 different isolates of P. salmonis and does not cross-react with several other fish pathogens, revealing a high specificity and sensitivity. The test also detects P. salmonis in kidney tissue of infected coho salmon with 98% correlation with the immunofluorescence assay. PMID- 12093038 TI - Subclinical herpesvirus infection in farmed turbot Scophthalmus maximus. AB - Subclinical infections with a herpesvirus were detected by light and electron microscopic examination of juvenile turbot collected during a national surveillance programme. Virions detected in the epidermis of skin and in the epithelium of gills had a morphology consistent with those of Herpesvirus scophthalmi described from turbot in the United Kingdom and Denmark. This is the first report of herpesvirus infection in turbot in Norway. PMID- 12093040 TI - Flumequine in Atlantic salmon Salmo salar: disposition in fish held in sea water versus fresh water. AB - 14C-labeled flumequine was administered as a single oral (5 mg kg(-1), 86 microCi kg(-1)) or intravenous (5 mg kg(-1), 82 microCi kg(-1)) dose to Atlantic salmon Salmo salar held in sea water or in fresh water. The absorption, tissue distribution and elimination were determined by means of liquid scintillation counting and whole-body autoradiography. The drug was rapidly absorbed and extensively distributed in all groups of fish. Radiolabeled compound was present in blood and muscle for more than 8 wk in the freshwater groups. In the seawater groups, however, no radioactivity was detected in the blood and muscle after 4 d and 2 wk, respectively. It was concluded that flumequine was eliminated at a substantially higher rate from Atlantic salmon in sea water than in fresh water. PMID- 12093041 TI - Cimetidine enhances the plasma praziquantel concentration and treatment efficacy against Microcotyle sebastis in cultured rockfish Sebastes schlegeli. AB - The effect of cimetidine on the praziquantel concentration in the blood of the rockfish Sebastes schlegeli and the consequent effect on the treatment efficacy against Microcotyle sebastis were investigated. Fish were divided into 7 groups and orally administered praziquantel alone (200 and 100 mg kg(-1) body weight [BW]) or in combination with cimetidine (in doses of 200, 100 or 50 mg kg(-1) BW cimetidine with a praziquantel dose of 100 mg kg(-1) BW). The fish in the sixth group were coadministered 50 mg praziquantel and 200 mg cimetidine kg(-1) BW. The fish in the control group were administered only saline. At 24 h post-treatment, the plasma was analyzed for praziquantel by reversed-phase high-performance liquid chromatography (RP-HPLC) using diazepam as the internal standard, and the gills were examined to confirm the effectiveness of each treatment. The praziquantel concentration in plasma of fish administered 100 mg praziquantel + 200 mg cimetidine kg(-1) BW was not significantly different from that of fish treated with 200 mg praziquantel kg(-1) BW and was significantly (p < 0.05) higher (about 2 times) than that of fish administered 100 mg praziquantel kg(-1) BW. The group of fish administered 50 mg praziquantel + 200 mg cimetidine kg(-1) BW showed a similar plasma praziquantel concentration to that in the fish treated with 100 mg praziquantel kg(-1) BW. The treatment efficacies of the groups of fish coadministered 100 mg praziquantel kg(-1) BW and various concentrations of cimetidine (200, 100 and 50 mg kg(-1) BW) were not significantly different from that of the group of fish administered 200 mg praziquantel kg(-1) BW, but were significantly higher than those of the groups of fish fed 100 mg praziquantel kg( 1) BW alone or coadministered 50 mg praziquantel + 200 mg cimetidine kg(-1) BW. PMID- 12093042 TI - Factors influencing the distribution of Myxobolus cerebralis, the causative agent of whirling disease, in the Cache la Poudre River, Colorado. AB - Oligochaetes, triactinomyxons (TAMs), and age-0 trout were sampled in the upper Cache la Poudre River, Colorado, to determine the distribution of Myxobolus cerebralis during 1997 and 1998. Densities of the intermediate host, the oligochaete Tubifex tubifex, were 3.5 orders of magnitude higher in the M. cerebralis-infected Poudre Rearing Unit (PRU) trout rearing ponds than at any of the river sampling reaches. Oligochaetes, including T. tubifex, were rare in the river (1 oligochaete m(-2)), except in a few stream side alcoves and eddies (50 oligochaete m(-2)). Species composition of oligochaetes in the river reaches was more diverse than in the PRU. Tubifex tubifex constituted 50% or less of the oligochaete community in the river and 98% in the PRU. Infection rates of T tubifex were 1% in the area above the PRU, 2% in the PRU, and 6% below the PRU. An increased M. cerebralis intensity of infection in age-0 trout below the PRU could not be attributed entirely to the high numbers of TAMs in its effluent (3.7 TAMs l(-1)). Low densities of TAMs ranging from 0 to 0.2 TAMs l(-1) were found in the river reaches, yet nearly all of the age-0 trout were infected soon after emergence. This suggests that very few TAMs, as measured by filtration, need be present in the water column to bring about infection in the majority of trout present. This also indicates that the parasite can persist and potentially cause reduced juvenile trout recruitment in cold, oligotrophic, sediment poor, high gradient streams. PMID- 12093044 TI - Monoclonal antibodies specific to yellow-head virus (YHV) of Penaeus monodon. AB - Monoclonal antibodies specific to 22, 67 and 135 kDa proteins of yellow-head virus (YHV) were produced from a mouse immunized with partially purified YHV isolated from the haemolymph of experimentally YHV-infected Penaeus monodon. Four groups of monoclonal antibodies were identified. One group of antibodies bound only to native protein of YHV while the others were specific to 135, 67 and 22 kDa proteins in both native and denatured forms. All antibodies could be used to detect YHV infection by means of dot blot and immunohistochemistry. However, antibodies specific to the 22 kDa protein gave the best immunohistochemistry results in terms of intensity and sharpness of staining. PMID- 12093043 TI - Development of an 18S rRNA gene-targeted PCR-based diagnostic for the blue crab parasite Hematodinium sp.. AB - The 18S rRNA gene from Hematodinum sp., a parasitic dinoflagellate that infects blue crabs, was amplified, cloned, and sequenced. The sequence showed a high similarity (95% at the nucleotide level) to sequences obtained from other dinoflagellate species, including both free-living and symbiotic species. Sequence similarity was much lower when compared with parasites of other marine invertebrates with similar life histories and with the 18S rRNA gene from the blue crab. Based on comparison of sequence alignments between Hematodinium, other dinoflagellate species, protozoan pathogens of oysters, and blue crab 18S rRNA gene sequences, 2 sets of PCR primers that specifically amplified fragments of the Hematodinium 18S rRNA gene were developed and tested. One of these primer sets (Hemat-F-1487 and Hemat-R-1654) amplified a 187 bp fragment that could be used routinely as a diagnostic test for the presence of Hematodinium in hemolymph from blue crabs. This fragment was consistently amplified from genomic DNA extracted from hemolymph of Hematodinium infected blue crabs. Comparison between the PCR technique and standard histological examination indicated that the PCR technique was reliable and provided 1000 times more sensitivity than the histological methods. The sensitivity of the PCR diagnostic was estimated to be one parasite cell among 300,000 crab hemocytes. Preliminary studies using the PCR diagnostic technique suggest that Hematodinium sp. is absent in crabs collected from waters with low salinity (5 to 10 ppt), but common in crabs from higher salinity environments in estuarine waters from southeastern Georgia (USA). PMID- 12093045 TI - Pericarditis and myocarditis in farmed Atlantic halibut Hippoglossus hippoglossus. AB - Farming of Atlantic halibut Hippoglossus hippoglossus is a growing industry in Norway. This study comprises the first description of heart morphology in fish from one Atlantic halibut farm. Fish with no clinical signs of disease were randomly sampled (8 to 12 mo old, weight 1 to 50 g). The hearts were studied by light and electron microscopy and samples were taken from the kidney and pericardial cavity for bacteriological examination. Slight to severe changes were found in most hearts at all samplings. The most serious pericardial and myocardial changes were seen in larger fish (15 to 50 g). In some of the hearts the affected pericardium formed multifocal protrusions. Eight fish had an adhesion between the apex of the ventricle and the parietal wall of the pericardial cavity. Results of bacteriology and immunohistochemistry investigations for common disease agents were negative. PMID- 12093046 TI - TIME has come to have a closer look at the management of cardiovascular disease in the elderly. PMID- 12093047 TI - Echocardiography or Nt-proANP in post-myocardial infarction patients: is one enough? PMID- 12093048 TI - Coronary angiography and revascularization for acute coronary syndromes without ST elevation: the next challenge. PMID- 12093049 TI - No room for radiant dreams in the real world. PMID- 12093050 TI - Amiodarone: pearl or peril? PMID- 12093051 TI - Intermittent claudication: how should we react to this symptom? PMID- 12093052 TI - Electrical and mechanical support in advanced heart failure. Rationale and feasibility of a combined management strategy. PMID- 12093053 TI - Prognostic value of two-dimensional echocardiography and N-terminal proatrial natriuretic peptide following an acute myocardial infarction. Assessment of baseline values (2-7 days) and changes at 3 months in patients with a preserved systolic function. AB - AIMS: The purpose of this prospective, observational study was to evaluate the relationship of left ventricular volumes, systolic function and plasma N-terminal proatrial natriuretic peptide (Nt-proANP) to cardiac morbidity and mortality in post-myocardial infarction patients with left ventricular ejection fraction > or =40%. METHODS AND RESULTS: Two-dimensional echocardiographic recordings and Nt proANP measurements were obtained in 834 patients who survived acute myocardial infarction. Patients were examined at 2-7 days and 3 months after the index infarction and followed up for 24 months. All measurements of left ventricular volumes, ejection fraction and Nt-proANP were performed in core laboratories. During follow-up 102 patients sustained one or more incidents of the combined primary end-point: cardiac death (n=11), recurrent infarction (n=55) or heart failure requiring hospitalization or treatment with an ACE inhibitor and a diuretic (n=52). Using Cox proportional hazards model, baseline Nt-proANP predicted these events (chi-square 25.3, P<0.0001), while baseline echo volumes and ejection fraction did not. During the subsequent 3-24 month period, 51 patients suffered a primary end-point: cardiac death (n=9), recurrent infarction (n=29), heart failure (n=21). An increase in left ventricular end-systolic volume was the strongest predictor for adverse events (chi-square 19.1, P<0.0001), especially for heart failure. Individual changes in Nt-proANP did not predict cardiac events, whereas both echocardiographic variables and Nt-proANP measured at 3 months had a prognostic impact on subsequent cardiac events (3-24 months). CONCLUSIONS: In post-myocardial infarction patients with preserved left ventricular function (left ventricular ejection fraction > or =40%) baseline Nt proANP, but not echocardiographic left ventricular volumes predicted adverse cardiac events. Early changes in left ventricular volumes and ejection fraction from baseline to 3 months had a further prognostic impact on subsequent events (3 24 months). PMID- 12093054 TI - First national survey on management strategies in non ST-elevation acute ischaemic syndromes in Argentina. Results of the STRATEG-SIA study. AB - AIMS: To assess the management strategies applied in non-ST elevation acute ischaemic syndromes in Argentina, the factors influencing the choice of treatment, and their relationship to short- and long-term (1 year) patient outcomes. METHODS AND RESULTS: We conducted a 1 month, prospective, population based survey in 77 hospitals (all over the country). We recruited 492 patients (age 63.9+/-11.7 years, male sex 68.3%, and 59.8% acute ischaemic ECG changes). Subjects were stratified according to the AHCPR classification as: high risk 62.2%, intermediate 25.0% and low 12.8%. At 1 year, the rate of death or myocardial infarction according to risk category and invasive procedures employed were: high risk (angioplasty 5.4% vs coronary artery bypass grafting 12.1% vs medical treatment 17.2%; P=0.001), intermediate risk (angioplasty 5.7% vs coronary artery bypass grafting 12.5% vs medical treatment 4.7%, P=ns), and low risk (angioplasty 10.0% vs coronary artery bypass grafting 15.2% vs medical treatment 1.9%; P<0.001). In the overall population, the 1 year event rate was not significantly different between the invasive and medical treatment groups (11.5% vs 7.2%, P=0.09). CONCLUSIONS: A routine, unselected invasive approach in non-ST elevation acute ischaemic syndromes in Argentina is associated with no apparent improvement of patients' outcome. PMID- 12093055 TI - Exercise-electrocardiography and/or pharmacological stress echocardiography for non-invasive risk stratification early after uncomplicated myocardial infarction. A prospective international large scale multicentre study. AB - AIMS: The aim of the present study was to assess the relative prognostic value of clinical variables, the exercise electrocardiography test and the pharmacological stress echocardiography test either with dipyridamole or dobutamine early after a first uncomplicated acute myocardial infarction in a large, multicentre, prospective study. METHODS AND RESULTS: Seven hundred and fifty-nine in-hospital patients (age=56+/-10 years) with a recent and first clinical uncomplicated myocardial infarction, with baseline echocardiographic findings of satisfactory quality, an interpretable ECG and able to exercise underwent a resting 2D echocardiogram, a pharmacological stress test with either dipyridamole or dobutamine and an exercise electrocardiography test at a mean of 10 days from the infarction; they were followed-up for a median of 10 months. During the follow up, there were 13 deaths, 23 non-fatal myocardial infarctions and 59 re hospitalizations for unstable angina. When all spontaneous events were considered, with multivariate analysis, the difference between the wall motion score index at rest and peak stress (delta wall motion score index), and exercise duration were independent predictors of future spontaneous events (relative risk 7.2; 95% CI=2.73-19.1; P=0.000; relative risk 1.1, 95% CI=1.02-1.18; P=0.008, respectively). Kaplan-Meier survival estimates showed a better outcome for those patients with a negative pharmacological stress echocardiography test compared to patients with low dose positivity (94.7 vs 74.8%, P=0.000). CONCLUSION: Stress echocardiography tests provide stronger information than historical and exercise electrocardiography test variables. Pharmacological echocardiography as well as the exercise ECG is able to predict all spontaneously occurring events when the presence as well as the timing, severity, and extension of stress-induced wall motion abnormalities are considered. PMID- 12093056 TI - Routine intracoronary beta-irradiation. Acute and one year outcome in patients at high risk for recurrence of stenosis. AB - AIMS: Intracoronary radiation is a promising therapy potentially reducing restenosis following catheter-based interventions. Currently, only limited data on this treatment are available. The feasibility and outcome in daily routine practice, however, is unknown. METHODS AND RESULTS: In 100 consecutive patients, intracoronary beta-radiation was performed with a (90)Strontium system (Novoste Beta-Cathtrade mark) following angioplasty. Predominantly complex (73% type B2 and C) and long lesions (length 24.3+/-15.3 mm) were included (37% de novo, 19% restenotic and 44% in-stent restenotic lesions). Radiation success was 100%. Mean prescribed dose was 19.8+/-2.5 Gy. A pullback procedure was performed in 19% lesions. Geographic miss occurred in 8% lesions. Periprocedural thrombus formation occurred in four lesions, dissection in nine lesions. During hospital stay, no death, acute myocardial infarction, or repeat revascularization was observed. Major adverse cardiac events occurred predominantly between 6 and 12 months after the index procedure with major adverse cardiac event-free survival of 66% at 12 months (one death, 10 Q-wave myocardial infarctions, 23 target vessel revascularizations; ranked for worst event). CONCLUSION: Routine catheter based intracoronary beta-radiation therapy after angioplasty is safe and feasible with a high acute procedural success. The clinical 1-year follow-up showed delayed occurrence of major adverse cardiac events between 6 and 12 months after the index procedure. PMID- 12093057 TI - Percutaneous implantation of a valve in the descending aorta in lambs. AB - AIMS: We assessed the feasibility of percutaneous implantation of a valve in the descending aorta and its function in systemic pressures. METHODS AND RESULTS: A biological valve harvested from a bovine jugular vein was sutured into a stent. After the creation of aortic insufficiency, the valved stent was percutaneously implanted into eight lambs divided into two groups depending on the severity of the insufficiency created. Haemodynamic and angiographic evaluations were carried out. Anatomical evaluation was finally performed. Aortic insufficiency was created: four lambs had mild insufficiency and four massive insufficiency. A valve was successfully implanted in all and were functioning perfectly in the early follow-up. Despite this competence, animals with massive insufficiency died within 24 h following implantation. None of the animals with mild insufficiency died. The valve was functioning perfectly in the first 2 months following the implantation, but became incompetent after spontaneous healing of the wound of the aortic valve. CONCLUSION: Percutaneously implanted valves in the descending aorta of lambs with aortic insufficiency function well in the early follow-up. This technique might become an interesting alternative to the standard approach in patients in whom perioperative risk is high. PMID- 12093058 TI - Amiodarone reduces procedures and costs related to atrial fibrillation in a controlled clinical trial. AB - BACKGROUND: Atrial fibrillation is the most common sustained cardiac arrhythmia, and engenders significant health care costs. The impact of various treatment options for atrial fibrillation on hospital costs has not been evaluated in a randomized trial. METHODS: We analysed 1-year follow-up data on 392 patients randomized to low dose amiodarone (200 mg. day(-1)) or alternative first-line therapy (sotalol or propafenone) in a multicentre trial (Canadian Trial of Atrial Fibrillation, CTAF). RESULTS: Patients in the amiodarone group had fewer electrical cardioversions (65 vs 109 for patients in the sotalol/propafenone group, P<0.0001), and pacemaker insertions (4 vs 11, P=0.07). The average amiodarone patient spent fewer days in hospital (0.47 vs 0.97, P=0.01), and incurred lower costs ($532 vs $898, P=0.03), for admissions where atrial fibrillation was the admitting diagnosis. Average total hospital costs per patient for all admissions, as well as average combined hospital and physician costs per patient, showed wide variations within the treatment arms and were not significantly different between groups. CONCLUSION: For patients in whom antiarrhythmic drug therapy is indicated, low dose amiodarone significantly reduces atrial fibrillation-related costs by reducing the number of atrial fibrillation-related procedures. PMID- 12093059 TI - Sulodexide in the treatment of intermittent claudication. Results of a randomized, double-blind, multicentre, placebo-controlled study. AB - AIMS: Patients with peripheral arterial obstructive disease require treatment to prevent major cardiovascular events and to relieve intermittent claudication. The walking performance of peripheral arterial obstructive disease patients was used to evaluate the usefulness of sulodexide, a glycosaminoglycan containing fast moving heparin and dermatan sulphate. METHODS AND RESULTS: A randomized, multicentre, double-blind, placebo-controlled study was performed in 286 patients with Leriche-Fontaine stage II peripheral arterial obstructive disease. Patients received placebo (n=143) or sulodexide (n=143) for 27 weeks. The primary end point was the doubling of the pain-free walking distance at the end of treatment, and this was achieved by 23.8% of patients treated with sulodexide and 9.1% of those on placebo (P=0.001). The pain-free walking distance increased on average (+/-SE) by 83.2+/-8.6 m (+64.7% from baseline) with sulodexide and 36.7+/-6.2 m (+29.9% from baseline) with placebo (P=0.001). The maximum walking distance increased by 142.3+/-15.8 m (+76.0% from baseline) and 54.5+/-8.4 m (+27.9% from baseline) (P<0.001), respectively. Results for patients with type II diabetes were similar to those for non-diabetic patients. Plasma fibrinogen decreased with sulodexide, but increased with placebo. CONCLUSION: Sulodexide improved the walking ability of peripheral arterial obstructive disease patients to a significantly greater extent than placebo, with a concurrent significant decrease in fibrinogen. The treatment was well tolerated. PMID- 12093061 TI - Tissue engineering of small diameter vascular grafts. AB - Tissue engineering, using either polymer or biological based scaffolds, represents the newest approach to overcoming limitations of small diameter prosthetic vascular grafts. Their disadvantages include thromboembolism and thrombosis, anticoagulant related haemorrhage, compliance mismatch, neointimal hyperplasia, as well as aneurysm formation. This current review represents an overview about previous and contemporary studies in the field of artificial vascular conduits development regarding arterial and venous autografts, allografts, xenografts, alloplastic prostheses, and tissue engineering. PMID- 12093062 TI - A multiantioxidant supplementation reduces damage from ischaemia reperfusion in patients after lower torso ischaemia. A randomised trial. AB - BACKGROUND: open repair of intra-abdominal aortic aneurysm (AAA) is associated with lower torso ischaemia and reperfusion. OBJECTIVE: to examine the effect of antioxidants on the activation and sequestration of white blood cells and muscle injury during AAA repair. METHOD: forty-two patients undergoing elective infrarenal aneurysm repair, were randomised to either standard therapy (22 patients) or standard therapy with additional multiantioxidant supplementation (20 patients). Vitamin E and C, Allopurinol, N-acetylcysteine and mannitol was administered perioperatively. White blood cell count (WBC), serum creatine kinase, aspartateaminotransferase, lactate and lipofuscine were measured. RESULTS: WBC remained higher after reperfusion in the antioxidant group (p = 0.008). CK, ASAT and lipofuscine levels were significantly lower after reperfusion in the antioxidant group (p = 0.02, p = 0.018, p = 0.017). CONCLUSION: multi-antioxidant supplementation was associated with a reduction in serum CK and ASAT after AAA repair. This is likely due to a reduction in oxidative stress and a decreased leucocyte sequestration and activation. PMID- 12093063 TI - Impact of treatment on the quality of life in patients with critical limb ischaemia. AB - AIM: the purpose of this study was to assess the quality of life (QOL) in patients with critical lower limb ischaemia (CLI), and its modification after three types of therapeutical outcomes. MATERIALS AND METHODS: between January and December 2000 52 patients (36 male, mean age 68 years) treated for critical limb ischaemia (CLI) completed the Short Form (SF)-36 on admission and then 6 and 12 months later. Patients were divided in the 3 groups: Group I: revascularisation; Group II: major amputation; Group III: conservative treatment. Trends for each dimension over the follow-up and the effect of treatment group were assessed by an analysis of variance with repeated measurements. RESULTS: on admission, patients with CLI had a significantly worse health-related quality of life than a control population. Although some domains improved, some also deteriorated between baseline and 12 months and there was no significant difference between the three treatment groups. CONCLUSIONS: our results suggest that changes in dimensions of QOL obtained by limb revascularisation do not differ from those of other therapeutic approaches. PMID- 12093064 TI - Nottingham health profile scores predict the outcome and support aggressive revascularisation for critical ischaemia. AB - AIMS: to assess changes in health related quality of life (HRQoL) following peripheral arterial reconstruction for critical limb ischaemia (CLI). METHODS: sixty patients with CLI were prospectively evaluated with the Nottingham Health Profile (NHP) before and 3 and 12 months after arterial reconstruction. In addition, at 12 months, patients were asked if their expectations of the revascularisation had been met and whether they considered the surgical treatment had been worthwhile. RESULTS: primary amputations and mortality were 5% and 3% and after 12 months 12% and 12% respectively. Three months after surgery scores on the pain and sleep sections of the NHP had improved significantly (p < 0.05). These improvements were maintained at 12 months. CONCLUSION: revascularisation for CLI improves HRQoL and pre-operative health perceptions are related to surgical outcome. PMID- 12093065 TI - Pre-operative hand-held Doppler run-off score can be used to stratify risk prior to infra-inguinal bypass surgery. AB - OBJECTIVES: to investigate the relationship between calf vessel run-off assessed by hand-held Doppler, graft patency and patient survival following infra-inguinal graft surgery. DESIGN: prospective cohort study of 258 consecutive patients undergoing infra-inguinal bypass grafts in one centre between 1995-99. MATERIALS AND METHODS: ankle Doppler auditory waveform characteristics were documented for patients considered for infra-inguinal bypass grafting. Doppler signals from the anterior tibial, posterior tibial and dorsalis pedis arteries were scored triphasic/biphasic (2), monophasic (1) or absent (0). A total Doppler run off score (0-6) was calculated. Following surgery graft surveillance was undertaken using duplex ultrasound at 6, 12, 26 and 52 weeks. Graft and patient survival were analysed using Cox regression analysis. RESULTS: overall primary assisted graft patency at one year was 80%. With an increasing Doppler score from 0 to 6, primary assisted graft patency steadily rose from 50% to 100% (p = 0.0002), accompanied by a steady fall in patient mortality from 50% to 5% (p = 0.0003). PMID- 12093066 TI - C-reactive protein is elevated in symptomatic compared with asymptomatic patients with carotid artery disease. AB - OBJECTIVES: to investigate the level of inflammatory markers between symptomatic and asymptomatic carotid stenosis patients. DESIGN: cross-sectional study. MATERIALS AND METHODS: a prospective study of 137 consecutive patients, admitted electively for carotid endarterectomy during 1997-2000, was conducted. 125 patients had cerebrovascular symptoms: either stroke (neurological deficit >24 h), Transient ischaemic attack (neurological deficit<24 h) or amaurosis fugax. Twelve patients were asymptomatic. A medical history and a fasting venous blood sample were taken from each patient around 6 weeks before surgery. The plasma concentrations of cholesterol and of inflammatory markers; (high sensitivity C reactive protein (hs-CRP), sICAM-1, sVCAM-1, sE-selectin) were determined. RESULTS: the concentration of hs-CRP in the symptomatic group (3.9 mg/L) was significantly higher than in the asymptomatic group (2.1 mg/L; p = 0.04). These concentrations were within normal range (<10 mg/L). sICAM-1, sVCAM-1, sE-selectin and total cholesterol concentrations were not different between the two groups. CONCLUSION: plasma hs-CRP was elevated in symptomatic compared to asymptomatic patients with carotid artery disease. High sensitivity C-reactive protein has been shown to be of prognostic value in a number of cardiovascular conditions and this study suggests it may be of value to identify patient at high risk of developing neurological deficits. PMID- 12093067 TI - Diagnostic performance of duplex ultrasonography in the detection of high grade internal carotid artery stenosis. AB - OBJECTIVES: to establish on a national basis whether the diagnostic accuracy of carotid duplex justifies carotid surgery without preoperative angiography. DESIGN: prospective national multicentre study with 10 participating university and county hospitals. MATERIAL AND METHODS: one hundred and thirty-four patients, aged 69+/-9 years, were subjected to routine carotid duplex ultrasonography and angiography. The influence of relevant factors on the relation between ultrasonographic and angiographic variables was evaluated using multiple regression analysis. The capacity of carotid ultrasonography to detect internal carotid artery (ICA) stenosis > or =80% was assessed by receiver operating characteristic analysis. RESULTS: the correlation between peak systolic velocity in ICA (PSV(ICA)) and the angiographic degree of stenosis was strong and significantly influenced only by the applied Doppler angle. Accordingly, the optimal PSV(ICA) cutpoint values for the diagnosis of ICA stenosis > or =80% (ECST method) differed substantially (2.1 and 3.2 m/s) between the two considered angle ranges (0-49 degrees and 50-62 degrees ), the ability to identify high grade ICA stenosis being significantly better at small Doppler angles (0-49 degrees ). CONCLUSION: ultrasonographic duplex technique identifies high grade ICA stenosis with a high degree of accuracy, which can be further improved by the application of small Doppler angles and the use of angle range specific PSV(ICA) cut-off points. PMID- 12093068 TI - Asymptomatic carotid stenosis in patients on medical treatment alone. AB - OBJECTIVE: the aim of this study was to investigate the effect of currently recommended medical treatment (MT) on changes in carotid stenosis in a group of asymptomatic patients taken from the Asymptomatic Carotid Surgery Trial (ACST). METHOD: collaborators in ACST were given information on MT for stroke prevention (including antiplatelet agents, lipid-lowering drugs, diabetic and hypertension control). Patients underwent clinical examination and duplex scanning at entry, 4 months following randomisation and annually thereafter. The cohort of patients studied were those randomised to MT with complete follow up duplex datasets at four years (n=219). None had undergone carotid endarterectomy (CEA) or developed ipsilateral carotid symptoms. RESULTS: there was no change in median carotid stenosis over four years (baseline 79% (IQR 10%) and 4 year median 79% (IQR 10%)) a median difference of 0 with Q1=-5 and Q3=+5 (p=0.98 Wilcoxon one sample test), whilst in many patients' stenoses progressed and regressed during this time. No individual MT variable correlated with stenosis progression or regression. CONCLUSION: in this group of ACST patients on MT, mean carotid stenosis was unchanged over 4 years. Individual patients' stenoses progressed (and regressed) without symptoms occurring. An increase in stenosis should not be the sole basis for deciding to operate on an asymptomatic patient. PMID- 12093069 TI - Adequate vascular training opportunities can be provided without compromising patient care. AB - OBJECTIVES: to review the results of index operations in vascular surgery and to evaluate the impact of a specialist vascular training programme on patient outcome. METHODS: we undertook a 5-year (January 1995-December 1999) review of the weekly-collected mortality and morbidity data. The total number, 30-day mortality and stroke rate of all index operations i.e. AAA repairs (ruptured and elective), carotid operations and infra-inguinal bypasses (above and below knee, elective and emergency) was recorded. The number of operations performed by trainees under supervision was recorded. RESULTS: in the 5-year period 991 index operation were done of which 738 (74%) were done by trainees. Operations done by trainees were supervised by a consultant in 82% of cases with no significant effect on death or stroke rates. Overall vascular trainees performed 75% of the index operations of which 82% were supervised. CONCLUSIONS: trainees under supervision performed three out of four index operations. The mortality and morbidity of index operations in our unit compared well with accepted best mortality and morbidity figures. Our unit provides good training opportunities for vascular trainees whilst maintaining satisfactory standards of patient care. PMID- 12093071 TI - Routine use of intravascular ultrasound for endovascular aneurysm repair: angiography is not necessary. AB - INTRODUCTION: to assess the outcome of endovascular aortic aneurysm repair (EVAR) using intravascular ultrasound (IVUS) without angiography. MATERIALS/METHODS: eighty consecutive patients (median age 69 years (range 25-90): male 72 (90%), female 8 (10%)) underwent endovascular aneurysm repair (AAA 68 (85%), TAA 12 (15%)) using either angiography in 31/80 patients (39%) or IVUS in 49/80 patients (61%) in accordance to the surgeons preference. RESULTS: hospital mortality was 2/80 (3%), 1/68 for AAA (2%), 1/12 for TAA (8%), 2/31 for angiography (7%), and 0/49 for IVUS (0.0%: NS). Median quantity of contrast medium was 190 ml (range: 20-350) for angiography versus 0 ml for IVUS (p<0.01). Median X-ray exposure time 24 min (range 9-65 min) versus 8 min (range 0-60 min) for IVUS (p<0.05). No coverage of renal or suprarenal artery orifices occurred in either group. Conversion to open surgery was necessary in 4/80 patients (5%), 1/31 for angiography (3%) and 3/49 patients for IVUS (6%: NS). Early endoleaks were observed in 13/80 patients (16%): 8/31 patients for angiography (26%) versus 5/49 for IVUS (10%: p<0.05): 5/13 endoleaks resolved spontaneously (39%) whereas 8/13 (61%) required additional procedures. CONCLUSIONS: IVUS is a reliable tool for EVAR. In most cases, perprocedural angiography is not necessary. PMID- 12093070 TI - Endovascular repair with bifurcated stent-grafts under local anaesthesia to improve outcome of ruptured aortoiliac aneurysms. AB - INTRODUCTION: acute haemodynamic changes and/or loss of abdominal muscle tone can occur during induction of general anaesthesia and may be the Achilles' tendon in endovascular aneurysm repair (EVAR) of ruptured aortoiliac aneurysms (rAIA). The purpose of this study was to evaluate the use of local anaesthesia (LA) for EVAR to overcome these limitations. METHODS: twenty-one consecutive patients with rAIA are included in this study. Twenty patients underwent EVAR under LA, and 1 patient was treated under general anaesthesia. Haemodynamics were stabilised during assessment of EVAR feasibility by CT-scan and during the procedure itself by controlled hypotension (MAP 50-60 mmHg) and moderate fluid resuscitation. RESULTS: median procedure time was 120 min. Haemodynamics remained stable in all but 3 patients who required transfemoral balloon occlusion of the supra-renal aorta. Perioperative intubation was necessary in 5 patients because of respiratory distress (n=3), or retroperitoneal access (n=2). Temporary deterioration of renal function occurred in 6 patients, with 2 requiring hemofiltration. CT-scan confirmed sealing of the rAIA in all patients at discharge. 30-day mortality was 9.5% (2 deaths). In the median follow-up of 19 months, there were no deaths, but 3 endovascularre-interventions, 1 crossover femoro-femoral bypass, and 1 open surgical graft repair. DISCUSSION: our series is the first to show that EVAR for rAIA can be safely performed under LA. This approach allows implantation of commercially available bifurcated SG and improves patient outcome. PMID- 12093072 TI - Alteration of elastin, collagen and their cross-links in abdominal aortic aneurysms. AB - OBJECTIVES: although the mechanism of arterial dilation and aneurysm development has not been clarified, the degradation of elastin and collagen plays undoubtedly a critical role. We evaluated the elastin and collagen content through the detection of their cross-links in aneurysmal and non-aneurysmal abdominal aortic walls. MATERIALS AND METHODS: in 26 human abdominal aortic aneurysm specimens obtained during surgery and in 24 autopsy control samples of non-aneurysmal abdominal aorta the tissue content of elastin and collagen cross-links were measured by HPLC. Collagen was also detected by evaluating two characteristic amino acids, 4-hydroxyproline (4-hypro) with a colorimetric method and 5 hydroxylysine (5-hylys) by gas chromatography. RESULTS: significantly fewer elastin cross-links were found in aneurysm samples compared to controls (desmosines and isodesmosines: 90% reduction; p<0.01). The opposite was true for pyridinoline collagen cross-links (350% increase) and deoxypyridinolines (100% increase, p=0.01). Tissue content of 5-hylys, 4-hypro and total amino acids were reduced significantly by 50% in aneurysmal samples. CONCLUSIONS: beside confirming decreased elastin content in aneurysmal walls, these results show a concurrent increase of collagen cross-links. Since total collagen markers were decreased (decreased 4-hypro and 5-hylys) it is reasonable to suggest that in aneurysmal aortic walls old collagen accumulates cross-links while new collagen biosynthesis is somehow defective. PMID- 12093074 TI - Abdominal aortic aneurysm in association with horseshoe kidney. PMID- 12093073 TI - Generation of reactive oxygen metabolites by the varicose vein wall. AB - OBJECTIVES: to evaluate the content of lipid peroxidation products (expressed by the concentration of thiobarbituric acid-reactive substances; TBARS), the content of myeloperoxidase (MPO), and the localisation of xanthine oxidase (XO) in varicose veins (vv), varicose veins with superficial thrombophlebitis and unchanged saphenous veins. METHODS: varicose saphenous veins, varicose veins with superficial thrombophlebitis and normal saphenous veins obtained during varicose vein surgery on 36 patients as well as healthy saphenous veins from cadaver organ donors (control). Homogenates were prepared in which TBARS concentration and MPO content were determined. Immunohistochemical staining to detect XO was also performed. RESULTS: the highest concentration of TBARS occurred in vv with superficial thrombophlebitis, the lowest in donor vein. The highest content of MPO was observed in vv and slightly lower - in varicose veins with thrombophlebitis. A positive reaction for XO was seen in vv wall endothelium. Specimens of vv with thrombophlebitis revealed strong, intense staining in endothelium as well as in vasa vasorum. CONCLUSIONS: varicose veins, especially those complicated with superficial thrombophlebitis revealed increased free radical generation. Its sources might be neutrophils, and in vv complicated with superficial thrombophlebitis-xanthine oxidase. PMID- 12093075 TI - A safe method of retrograde passage of fogarty embolectomy catheter through difficult iliac arteries. PMID- 12093076 TI - The occlusion of an internal iliac aneurysm by the trans-catheter injection of thrombin. PMID- 12093077 TI - The effect of electrical stimulation of the calf muscle and compression stocking on venous blood flow velocity. PMID- 12093078 TI - Amyotrophic lateral sclerosis/parkinsonism dementia complex: transgenic mice provide insights into mechanisms underlying a common tauopathy in an ethnic minority on Guam. AB - Intracytoplasmic filamentous tau inclusions are neuropathological hallmarks of amyotrophic lateral sclerosis/parkinsonism-dementia complex (ALS/PDC) of Guam and the defining lesions of other neurodegenerative disorders known as tauopathies. Here we review current insights into the cell and molecular neuropathology of ALS/PDC, a common tauopathy in the Chamorro population on Guam. We also summarize recent advances in understanding this disorder through studies of transgenic (Tg) mouse models of this tauopathy. Briefly, overexpression of human tau isoforms in the central nervous system of Tg mice resulted in a neurodegenerative tauopathy with a phenotype similar to ALS/PDC. Specifically, argyrophilic, congophilic, and tau immunoreactive inclusions accumulated with age in cortical and brainstem neurons of these mice, but they were most abundant in spinal cord neurons, and the inclusions contained 10- to 20-nm tau-positive straight filaments. There also was extensive gliosis in spinal cord associated with axonal degeneration in the ventral roots, while remaining axons in spinal nerves showed a loss of microtubules and reduced fast axonal transport. With advancing age, these Tg mice showed increasing motor weakness, and this was accompanied by a progressive increase in the phosphorylation and insolubility of brain and spinal cord tau proteins. Thus, tau Tg mice recapitulate key phenotypic features of ALS/PDC neuropathology in an ethnic minority on Guam, and these animal models provide new opportunities to discover novel therapies for this and related tauopathies. PMID- 12093080 TI - Long-term injured purkinje cells are competent for terminal arbor growth, but remain unable to sustain stem axon regeneration. AB - Long-distance axon regeneration requires the activation of a specific set of neuronal growth-associated genes. Adult Purkinje cells fail to upregulate these molecules in response to axotomy and show extremely weak regenerative properties. Nevertheless, starting from several months after injury, transected Purkinje axons undergo spontaneous sprouting. Here, we asked whether long-term injured Purkinje cells acquire novel intrinsic growth properties that enable them to upregulate growth-associated genes and sustain axon regeneration. To test this hypothesis, we examined axon growth and cell body changes in adult rat Purkinje neurons following axotomy and implantation of embryonic neocortical tissue or Schwann cells into the injury track. Purkinje cells that survived over 6 months after injury/transplantation displayed profuse sprouting in the injured cerebellum and developed extensive networks of terminal branches into embryonic neocortical grafts. In addition, severed Purkinje axons exposed to these transplants 6 months after injury grew faster than their counterparts confronted with the same environment immediately after axotomy. Nevertheless, long-term injured Purkinje cells failed to regenerate stem neurites into Schwann cell grafts, and, under all experimental conditions, they did not upregulate growth associated molecules, including c-Jun, GAP-43, SNAP-25, and NADPH-diaphorase. These results indicate that the long-term injured Purkinje cells remain unable to activate the gene program required to sustain axon regeneration and their plasticity is restricted to terminal arbor remodeling. We propose that the delayed growth of injured Purkinje cells reflects an adaptive phenomenon by which the severed axon stump develops a new terminal arbor searching for alternative connections with local partners. PMID- 12093079 TI - Dentate granule cells in reeler mutants and VLDLR and ApoER2 knockout mice. AB - We have studied the organization and cellular differentiation of dentate granule cells and their axons, the mossy fibers, in reeler mutant mice lacking reelin and in mutants lacking the reelin receptors very low density lipoprotein receptor (VLDLR) and apolipoprotein E receptor 2 (ApoER2). We show that granule cells in reeler mice do not form a densely packed granular layer, but are loosely distributed throughout the hilar region. Immunolabeling for calbindin and calretinin revealed that the sharp border between dentate granule cells and hilar mossy cells is completely lost in reeler mice. ApoER2/VLDLR double-knockout mice copy the reeler phenotype. Mice deficient only in VLDLR showed minor alterations of dentate organization; migration defects were more prominent in ApoER2 knockout mice. Tracing of the mossy fibers with Phaseolus vulgaris leukoagglutinin and calbindin immunolabeling revealed an irregular broad projection in reeler mice and ApoER2/VLDLR double knockouts, likely caused by the irregular wide distribution of granule cell somata. Mutants lacking only one of the lipoprotein receptors showed only minor changes in the mossy fiber projection. In all mutants, mossy fibers respected the CA3-CA1 border. Retrograde labeling with DiI showed that malpositioned granule cells also projected as normal to the CA3 region. These results indicate that ( 1 ) reelin signaling via ApoER2 and VLDLR is required for the normal positioning of dentate granule cells and (2) the reelin signaling pathway is not involved in pathfinding and target recognition of granule cell axons. PMID- 12093081 TI - BMP-7 and excess glutamate: opposing effects on dendrite growth from cerebral cortical neurons in vitro. AB - Glutamate is an important regulator of dendrite development. During cerebral ischemia, however, there is massive release of glutamate reaching millimolar concentrations in the extracellular space. An early consequence of this excess glutamate is reduced dendrite growth. Bone morphogenetic protein-7 (BMP-7) a member of the transforming growth factor-beta (TGF-beta) superfamily has been demonstrated to enhance dendrite output from cerebral cortical and hippocampal neurons in vitro. However, it is not known whether BMP-7can prevent the reduced dendrite growth associated with excess glutamate or enhance dendrite growth after glutamate exposure. Therefore we quantified axon and primary, secondary, and total dendrite growth from embryonic mouse cortical neurons (E18) grown at low density in vitro in a chemically defined medium and exposed to glutamate (1 or 2 mM) for 48 h. Morphology and double immunolabeling (MAP2, NF-H) were used to identify cortical dendrites and axons after 3 DIV. In these short-term cultures, glutamate did not influence neuron survival. The addition of glutamate to cortical neurons, however, significantly attenuated dendrite output. This effect was mimicked by the addition of NMDA but not AMPA agonists and inhibited by the specific NMDA receptor antagonist MK-801. The reduction in dendrite growth mediated by excess glutamate was ameliorated by the administration of 30 or 100 ng/ml of BMP-7. In addition, when administered in a delayed fashion between 1 and 24 h after the initial glutamate exposure, BMP-7 was able to enhance dendrite growth, including primary dendrite number, primary dendrite length, and secondary dendritic branching. These findings demonstrate that BMP-7 can ameliorate reduced dendrite growth from cerebral cortical neurons associated with excess glutamate in vitro and are important because they may help explain why BMP-7 administration is associated with enhanced functional recovery in models of cerebral ischemia. PMID- 12093082 TI - The differential vulnerability of striatal projection neurons in 3-nitropropionic acid-treated rats does not match that typical of adult-onset Huntington's disease. AB - In adult-onset Huntington's disease (HD), striatal projection neurons are much more vulnerable than striatal interneurons, but even striatal projection neurons show differences in their vulnerability, with the striatal projection neurons projecting to the internal segment of the globus pallidus being the least vulnerable. Previous studies have shown that systemic chronic treatment with 3 nitropropionic acid (3NP), an inhibitor of succinate dehydrogenase, induces the preferential loss of striatal projection neurons over striatal interneurons that is characteristic of HD, which has been taken to support the hypothesis that the pathogenic defect in HD may involve impaired energy metabolism. We sought to determine whether the patterns of survival for striatal projection neurons in 4 month-old rats after chronic systemic 3NP treatment also resemble those in adult onset HD. We assessed the projection neuron survival using neuropeptide immunolabeling of striatal efferent fibers in striatal target areas and quantified the degree of fiber loss in the striatal target areas using computer assisted image analysis. We found that 3NP produced relatively equal loss of striatal fibers and terminals in the globus pallidus, substantia nigra, and entopeduncular nucleus, indicating a nondifferential vulnerability of striatal projection neurons to 3NP-induced impairment in energy metabolism. The results suggest that the 3NP rat model does not fully mimic adult-onset HD pathogenesis. PMID- 12093083 TI - Dose and promoter effects of adeno-associated viral vector for green fluorescent protein expression in the rat brain. AB - Previous studies demonstrated that the rat neuron-specific enolase (NSE) promoter is effective for transgene expression in the brain in a variety of adeno associated virus-2 vectors. This study evaluated the dose response and longer time course of this promoter and compared it to two cytomegalovirus/chicken beta actin hybrid (CBA) promoter-based systems. NSE promoter-driven green fluorescent protein (GFP)-expressing neurons were found at doses as low as 10(7) particles, with expression increasing in a dose-dependent manner over a 3.3-log range. Bicistronic expression of GFP via an internal ribosome entry site coupled to the NSE promoter was also dose dependent, although the potency was decreased by 3.4 fold. The number of GFP-expressing neurons was stable for at least 25 months. The CBA promoter increased the numbers of GFP-expressing cells versus the NSE promoter, although the expression pattern remained neuronal and persisted for at least 18 months. The CBA promoter permitted detection of cells distal to the injection site that had retrogradely transported the vector from their terminal areas. Incorporating the woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) into a CBA promoter vector induced greater expression levels in the hippocampus, as measured by stereological estimates of cell numbers and by Western blots, which demonstrated an 11-fold increase. Incorporation of the WPRE also improved transgene expression in primary neuronal cultures. The increased efficiency obtained with vector elements such as the CBA promoter and the WPRE may enhance the ability to genetically modify larger portions of the brain while requiring smaller doses and volumes. PMID- 12093085 TI - Upregulation of heat shock proteins rescues motoneurones from axotomy-induced cell death in neonatal rats. AB - Heat shock proteins (hsps) are induced in a variety of cells following periods of stress, where they promote cell survival. In this study, we examined the effect of upregulating hsp expression by treatment with BRX-220, a co-inducer of hsps, on the survival of injured motoneurones. Following sciatic nerve crush at birth, rat pups were treated daily with BRX-220. The expression of hsp70 and hsp90, motoneurone survival, and muscle function was examined at various intervals later and the number of functional motor units was assessed by in vivo isometric tension recordings. Fourteen days after injury, significantly more motoneurones survived in the BRX-220-treated group (39 +/- 2.8%) compared to the saline treated group (21 +/- 1.7%). Moreover, in the BRX-220-treated group no further loss of motoneurones occurred, so that at 10 weeks 42 +/- 2.1% of motoneurones survived compared to 15 +/- 0.6% in the untreated group. There were also more functional motor units in the hindlimb muscles of BRX-220-treated animals. In addition, treatment with BRX-220 resulted in a significant increase in the expression of hsp70 and hsp90 in glia and neurones. Thus, treatment with BRX-220, a co-inducer of hsps, protects motoneurones from axotomy-induced cell death. PMID- 12093086 TI - Demonstration of alpha-synuclein immunoreactivity in neuronal and glial cytoplasm in normal human brain tissue using proteinase K and formic acid pretreatment. AB - alpha-Synuclein (alphaS), a presynaptic nerve terminal protein, is now known to be a major component of neuronal and glial cytoplasmic inclusions in alpha synucleinopathies (Lewy body disease and multiple system atrophy). However, alphaS has not been identified in either neuronal or glial cytoplasm in formalin fixed, paraffin-embedded tissue sections from the normal human brain. Previous studies have shown that pretreatment with either proteinase K or formic acid enhances alphaS immunoreactivity. The aim of the present study was, therefore, to study the effects of pretreatment with proteinase K and formic acid on alphaS immunoreactivity in vibratome sections of brain tissue taken from normal human subjects. In addition to presynaptic staining, alphaS immunostaining was recognized in neuronal perikarya in the pretreated sections; this immunoreactivity was more intense in sections taken from the deeper layers of the cerebral neocortex, the CA2/3 region of the hippocampus, and the substantia nigra. This pattern of alphaS expression coincides with the distribution of intraneuronal inclusions in alphaS transgenic animals as well as in human autopsy tissue taken from patients with Lewy body disease. Furthermore, intense immunoreactivity was also found in the cytoplasm of astrocytes and oligodendrocytes throughout the brain. These findings suggest that a significant amount of alphaS is also present in the neuronal and glial cytoplasm in the normal human brain. PMID- 12093084 TI - Corticotropin-releasing hormone (CRH) downregulates the function of its receptor (CRF1) and induces CRF1 expression in hippocampal and cortical regions of the immature rat brain. AB - In addition to regulating the neuroendocrine stress response, corticotropin releasing hormone (CRH) has been implicated in both normal and pathological behavioral and cognitive responses to stress. CRH-expressing cells and their target neurons possessing CRH receptors (CRF1 and CRF2) are distributed throughout the limbic system, but little is known about the regulation of limbic CRH receptor function and expression, including regulation by the peptide itself. Because CRH is released from limbic neuronal terminals during stress, this regulation might play a crucial role in the mechanisms by which stress contributes to human neuropsychiatric conditions such as depression or posttraumatic stress disorder. Therefore, these studies tested the hypothesis that CRH binding to CRF1 influenced the levels and mRNA expression of this receptor in stress-associated limbic regions of immature rat. Binding capacities and mRNA levels of both CRF1 and CRF2 were determined at several time points after central CRH administration. CRH downregulated CRF1 binding in frontal cortex significantly by 4 h. This transient reduction (no longer evident at 8 h) was associated with rapid increase of CRF1 mRNA expression, persisting for >8 h. Enhanced CRF1 expression-with a different time course-occurred also in hippocampal CA3, but not in CA1 or amygdala, CRF2 binding and mRNA levels were not altered by CRH administration. To address the mechanisms by which CRH regulated CRF1, the specific contributions of ligand-receptor interactions and of the CRH-induced neuronal stimulation were examined. Neuronal excitation without occupation of CRF1 induced by kainic acid, resulted in no change of CRF1 binding capacity, and in modest induction of CRF1 mRNA expression. Furthermore, blocking the neuroexcitant effects of CRH (using pentobarbital) abolished the alterations in CRF1 binding and expression. These results indicate that CRF1 regulation involves both occupancy of this receptor by its ligand, as well as "downstream" cellular activation and suggest that stress-induced perturbation of CRH-CRF1 signaling may contribute to abnormal neuronal communication after some stressful situations. PMID- 12093087 TI - Traumatic brain injury in the immature mouse brain: characterization of regional vulnerability. AB - We characterized the regional and temporal patterns of neuronal injury and axonal degeneration after controlled cortical impact of moderate severity in mice at postnatal day 21. Animals were euthanized at 1, 3, or 7 days after injury or sham operation. The brains were removed and prepared for immunolocalization of neurons and microglia/macrophages or subjected to Fluoro-Jade and silver stains, indicators of irreversible neuronal cell injury and axonal degeneration. There was significant neuronal loss in both the ipsi- and the contralateral cortices, ipsilateral hippocampus, and ipsilateral thalamus by 7 days post injury compared to sham-operated animals. Activated microglia/macrophages were most prominent in regions of neuronal loss including the ipsilateral cortex, hippocampus, and thalamus. Neuronal injury, as evidenced by Fluoro-Jade labeling, was not apparent in sham-operated animals. In injured animals, labeling was identified in the ipsilateral cortex and hippocampus at 1 and 3 days post injury. Silver- and Fluoro-Jade-labeled degenerating axons were observed in the ipsilateral subcortical white matter by 1 day post injury, in the ipsilateral external capsule, caudate putamen, and contralateral subcortical white matter by 3 days post injury, and in the internal capsule, pyramidal tracts, and cerebellar peduncles by 7 days post injury. Our findings demonstrate that controlled cortical impact in the developing brain generates neuronal loss in both the ipsilateral and the contralateral cortex, a temporally distinct pattern of subcortical neuronal injury/death, and widespread white matter damage. These observations serve as an important baseline for studying human brain injury and optimizing therapies for the brain-injured child. PMID- 12093088 TI - Intranuclear aggregation of nonexpanded ataxin-3 in marinesco bodies of the nonhuman primate substantia nigra. AB - Marinesco bodies (MB) are intranuclear inclusion bodies predominantly found in melanin-pigmented neurons of the substantia nigra. MB are demonstrable not only in humans but also in nonhuman primates. In the present study MB of aged rhesus monkeys (Macaca mulatta; n = 15; mean age 16 years) and aged baboons (Papio anubis; n = 13; mean age 25 years) were examined immunohistochemically. MB were found to be immunoreactive for ubiquitin, a protein involved in initiation of proteasome-mediated proteolysis. We also demonstrate that MB in monkeys are intensely immunoreactive for the protein ataxin-3 as detected by using two monoclonal anti-ataxin-3 antibodies (1H9 and 2B6). The abnormally expanded form of this polyglutamine protein is known to be causally involved in spinocerebellar ataxia type 3 or Machado-Joseph disease. The monoclonal antibody 1C2 was employed to examine whether ataxin-3 in MB in monkeys contains such an abnormally expanded polyglutamine stretch. MB were consistently 1C2-immunonegative, indicating that they are composed of normal wild-type ataxin-3. In conclusion MB in nonhuman primates permit experimental examination of mechanisms involved in transnuclear localization, intranuclear aggregation, and ubiquitination of nonexpanded polyglutamine proteins. PMID- 12093089 TI - Sex differences in androgen receptor immunoreactivity in basal forebrain nuclei of elderly and Alzheimer patients. AB - The vertical limb of the diagonal band of Broca (VDB or Ch2) and the nucleus basalis of Meynert (NBM or Ch4) are major cholinergic nuclei of the human basal forebrain, a complex that is affected in Alzheimer's disease (AD). Sex hormones influence the function of these cholinergic neurons in animals and humans and we showed earlier that estrogen and androgen receptors (AR) are present in both the VDB and the NBM of young patients of 20-39 years of age. The aim of the present study was to investigate whether AR expression changes in relation to aging and AD. In both brain areas of male and female patients over the age of 56 nuclear staining had almost disappeared and cytoplasmic AR expression was decreased. This decrease was most pronounced in the VDB of men. In addition, the proportion of neurons showing cytoplasmic AR expression was higher in control aged women than in control aged men in both the VDB and the NBM. Surprisingly, cytoplasmic ARs were significantly decreased in the VDB and the NBM only in AD women and not in AD men. These observations suggest the possible involvement of androgens in the functional changes of the basal forebrain nuclei in aging and AD. PMID- 12093091 TI - Anatomical correlates of locomotor recovery following dorsal and ventral lesions of the rat spinal cord. AB - The present study was designed to relate functional locomotor outcome to the anatomical extent and localization of lesions in the rat spinal cord. We performed dorsal and ventral lesions of different severity in 36 adult rats. Lesion depth, spared total white matter, and spared ventrolateral funiculus were compared to the locomotor outcome, assessed by the BBB open-field locomotor score and the grid walk test. The results showed that the preservation of a small number of fibers in the ventral or lateral funiculus was related to stepping abilities and overground locomotion, whereas comparable tissue preservation in the dorsal funiculus resulted in complete paraplegia. The strongest relation to locomotor function was between the BBB score and the lesion depth as well as the BBB score and the spared white matter tissue in the region of the reticulospinal tract. Locomotion on the grid walk required sparing in the ventrolateral funiculus and additional sparing of the dorsolateral and dorsal funiculus, where the cortico- and rubrospinal tracts are located. PMID- 12093090 TI - Priming by muscle inflammation alters the response and vulnerability to axotomy induced damage of the rat facial motor nucleus. AB - To ascertain whether signaling due to peripheral inflammation affects motoneuron vulnerability, we examined in adult rats the reaction to axonal injury of facial motoneurons primed by muscle inflammation. In this double-hit paradigm, preconditioning was achieved by injections into the facial muscles of the T cell mitogen phytohemagglutinin, which was found in a previous study ( 11 ) to elicit a retrograde response in motoneurons. Facial nerve transection was used as test lesion. Intramuscular injections of saline prior to axotomy were used as control for lectin pretreatment. In rats pretreated with phytohemagglutinin injection, upregulation of the expression of the antiapoptotic bcl-2 gene, examined with in situ hybridization, was significantly higher in facial motoneurons at 2 days postaxotomy compared with saline-injected control cases. After repeated phytohemagglutinin injections followed by nerve transection, induction in facial motoneurons of nitric oxide synthase, revealed by histochemistry and immunohistochemistry, as well as activation of the surrounding microglia, was enhanced at 14 days postaxotomy with respect to the saline-treated control cases. At the same time point, no significant intergroup difference was detected in the intensity of astrocytic activation. At 1 month postaxotomy, stereological cell counts revealed that motoneuron loss was significantly greater in the cases pretreated with phytohemagglutinin than in the saline-treated cases. The data point out that the response of the facial motor nucleus to axonal damage is altered by previous exposure to peripheral inflammation and that such preconditioning stimulus enhances motoneuron vulnerability to nerve injury. PMID- 12093092 TI - A role for tumor necrosis factor alpha in death of dopaminergic neurons following neural transplantation. AB - Poor survival of transplanted dopaminergic (DA) neurons remains a serious obstacle to the success of cell replacement therapy as an alternative to the current treatments for Parkinson's disease (PD). We have examined the temporal release profile of an inflammatory cytokine, tumor necrosis factor-alpha (TNFalpha), following transplantation of fetal mesencephalic tissue into the rat striatum. The amounts of TNFalpha released in vivo when added to cultures of embryonic DA neurons, significantly reduced the survival of DA neurons in vitro, and this cell death could be prevented by the inclusion of an antibody to the TNFalpha receptor type 1. Inclusion of this antibody in cell suspensions during transplantation also increased the survival of transplanted fetal DA neurons by approximately 250%. Use of this therapeutic antibody approach may offer significant improvements to neural transplantation as a treatment for PD. PMID- 12093093 TI - beta-Amyloid neurotoxicity is exacerbated during glycolysis inhibition and mitochondrial impairment in the rat hippocampus in vivo and in isolated nerve terminals: implications for Alzheimer's disease. AB - Senile plaques composed mainly by beta-amyloid (Abeta) protein are one of the pathological hallmarks of Alzheimer's disease (AD). In vitro, Abeta and its active fragment 25-35 have been shown either to be directly neurotoxic or to exacerbate the damaging effect of other neurotoxic insults. However, the attempts to replicate Abeta neurotoxicity in vivo have yielded conflicting results. One of the most consistent alterations in AD is a reduced resting glucose utilization. Important evidence suggests that impairment of brain energy metabolism can lead to neuronal damage or facilitate the deleterious effects of some neurotoxic agents. In the present study we have investigated the influence of glycolysis inhibition induced by iodoacetate, and mitochondrial impairment induced by 3 nitropropionic acid (3-NP), in the toxicity of Abeta. We have studied Abeta neurotoxicity during energy deficiency both in vivo in the dentate gyrus of the hippocampal formation and in presynaptic terminals isolated from neocortex and hippocampus. Results show that during metabolic inhibition an enhanced vulnerability of hippocampal neurons to Abeta peptide toxicity occurs, probably resulting from decreased glucose metabolism and mitochondrial ATP production. Synaptosomal response to energy impairment and Abeta toxicity was evaluated by the MTT assay. Results suggest that synapses may be particularly sensitive to metabolic perturbation, which in turn exacerbates Abeta toxicity. The present data provide experimental support to the hypothesis that certain risk factors such as metabolic dysfunction and amyloid accumulation may interact to exacerbate AD, and that metabolic substrates such as pyruvate may play a role as a therapeutic tool. PMID- 12093095 TI - Genetically engineered GABA-producing cells demonstrate anticonvulsant effects and long-term transgene expression when transplanted into the central piriform cortex of rats. AB - Local application of GABA-potentiating agents can prevent or reduce the development and maintenance of behavioral seizures induced by limbic kindling in rats. Microinjection and lesion studies suggest that the transition zone between anterior and posterior piriform cortex (PC), termed here central PC, is a potential target for transplantation of GABA-producing cells. In the present study, we transplanted conditionally immortalized mouse cortical neurons, engineered with the GABA-synthesizing enzyme GAD(65), to the central PC of rats. Suspensions of 1.5 x 10(5) cells in 1 microl were transplanted bilaterally. Control animals received transplantation of beta-galactosidase (beta-gal) expressing cells. All rats were subsequently kindled through a chronically implanted electrode placed in the basolateral amygdala. The pre- and postkindling threshold currents for eliciting behavioral seizures were determined before and after kindling. We found the prekindling partial seizure threshold to be significantly increased by about 200% in the rats that received the GABA producing cells compared to rats receiving beta-gal-producing transplants. After kindling, the seizure threshold tended to be higher by 100% in rats that received GABA-producing cells, although the difference from controls was not statistically significant. GABA-producing transplants had no significant effect on the rate of amygdala kindling, but the latency to the first generalized seizure during kindling was significantly increased in animals receiving GABA-producing cells. The transplanted cells showed long-term GAD(65) expression as verified immunohistologically after termination of the experiments. The findings substantiate and extend previous findings that the central PC is part of the anatomical substrate that facilitates propagation from partial to generalized seizures. The data demonstrate that genetically engineered cells have the potential to raise seizure thresholds when transplanted to the central PC. PMID- 12093094 TI - Adenosine and purine nucleosides protect rat primary astrocytes from peroxynitrite-potentiated, glucose deprivation-induced death: preservation of intracellular ATP level. AB - Previously we have reported that immunostimulated astrocytes became highly vulnerable to glucose deprivation. In the present study we examined the effect of various kinds of nucleosides on the augmented death of glucose-deprived immunostimulated astrocytes. Preincubation with interferon-gamma (100 U/ml) and lipopolysaccharide (1 microg/ml) for 48 h and continuous exposure to glucose deprivation (4 h) significantly induced the lactate dehydrogenase (LDH) release, as a marker of cell injury or death, from astrocytes. The glucose deprivation induced augmented cell death in immunostimulated astrocytes was mimicked by exogenous peroxynitrite generator 3-morpholinosydnonimine (SIN-1). The increased death in immunostimulated or SIN-1-treated astrocytes deprived of glucose was blocked by adenosine and ATP. Other purine nucleos(t)ides, not pyrimidine nucleotides, also showed similar protective effects. Adenosine receptor agonist R(-)-N-(2-phenylisopropyl)-adenosine or N-cyclohexyladenosine did not alter the augmented cell death. Adenosine receptor antagonists 8-cyclopentyl-1,3 dipropylxanthine, xanthine amine congener or 3,7-dimethyl-1-propargylxanthine also did not reverse the protective effect of adenosine. Intracellular ATP levels rapidly decreased prior to the LDH release in glucose-deprived immunostimulated astrocytes. The loss of intracellular ATP was prevented by adenosine and other purine nucleotides. The present results suggest that adenosine and their metabolites may protect astrocytes from peroxynitrite-potentiated, glucose deprivation-induced death by serving as substrates for intracellular ATP generation. PMID- 12093096 TI - Protection of malonate-induced GABA but not dopamine loss by GABA transporter blockade in rat striatum. AB - Previous work has shown that overstimulation of GABA(A) receptors can potentiate neuronal cell damage during excitotoxic or metabolic stress in vitro and that GABA(A) antagonists or GABA transport blockers are neuroprotective under these situations. Malonate, a reversible succinate dehydrogenase/mitochondrial complex II inhibitor, is frequently used in animals to model cell loss in neurodegenerative diseases such as Parkinson's and Huntington's diseases. To determine if GABA transporter blockade during mitochondrial impairment can protect neurons in vivo as compared with in vitro studies, rats received a stereotaxic infusion of malonate (2 micromol) into the left striatum to induce a metabolic stress. The nonsubstrate GABA transport blocker, NO711 (20 nmol) was infused in some rats 30 min before and 3 h following malonate infusion. After 1 week, dopamine and GABA levels in the striata were measured. Malonate caused a significant loss of striatal dopamine and GABA. Blockade of the GABA transporter significantly attenuated GABA, but not dopamine loss. In contrast with several in vitro reports, GABA(A) receptors were not a downstream mediator of protection by NO711. Intrastriatal infusion of malonate (2 micromol) plus or minus the GABA(A) receptor agonist muscimol (1 micromol), the GABA(A) Cl- binding site antagonist picrotoxin (50 nmol) or the GABA(B) receptor antagonist saclofen (33 nmol) did not modify loss of striatal dopamine or GABA when examined 1 week following infusion. These data show that GABA transporter blockade during mitochondrial impairment in the striatum provides protection to GABAergic neurons. GABA transporter blockade, which is currently a pharmacological strategy for the treatment of epilepsy, may thus also be beneficial in the treatment of acute and chronic conditions involving energy inhibition such as stroke/ischemia or Huntington's disease. These findings also point to fundamental differences between immature and adult neurons in the downstream involvement of GABA receptors during metabolic insult. PMID- 12093097 TI - Induction and activation of protein kinase C delta in hippocampus and cortex after kainic acid treatment. AB - Various isoforms of protein kinase C (PKC), especially the novel PKC subtypes delta, epsilon, and the atypical subtype PKC zeta, are involved in delayed cell death. We studied the expression and late activation of the latter PKC isoforms in comparison with classic PKC alpha, beta, and gamma in the brains of rats exposed to systemic kainate injection. The expression of PKC delta mRNA was strikingly upregulated (13-fold) in the cortex and the CA1 and CA3 hippocampal regions on 1 day after kainate administration, whereas PKC zeta mRNA was only moderately increased (about 100%) in these three brain regions on day 2 following the drug. PKC epsilon mRNA was slightly increased only in the cortex on days 2 and 6, while the mRNA levels of the classic PKC subtypes (alpha, beta, and gamma) remained unchanged or decreased after the treatment. Immunoblotting analyses revealed that the level of PKC delta protein started to increase on day 1 after kainate and was significantly elevated on day 2 in both the membrane and cytosol fractions of cortex and hippocampus. PKC epsilon protein only showed a marginal increase and the level of PKC zeta protein remained unaltered in response to the treatment. Cortical and CA1-3 pyramidal neurons displayed strong immunoreactivity for PKC delta on days 1 and 2, and microglia on days 1, 2, and 4 after the drug. The results indicate that the expression of apoptosis-associated isoforms of PKC, most notably that of delta, but to lesser extent also that of epsilon and zeta, is increased during kainate-induced neuronal death. The predominant induction of PKC delta in neurons and microglia suggests that PKC delta could be the major mediator or modulator of apoptotic and inflammatory responses to excitotoxic insults. PMID- 12093098 TI - The frequency of inherited hydrocephalus is influenced by intrauterine factors in H-Tx rats. AB - H-Tx rats have fetal-onset inherited hydrocephalus. Linkage analysis has determined the genetics is complex, with at least three loci associated with hydrocephalus. In addition, maternal and/or intrauterine factors influence the frequency of expression. The aim of this study was to characterize nongenetic (epigenetic) factors that affect hydrocephalus in this strain. Groups of primiparous and multiparous females were used to breed fetuses for examination in utero. Multiparous females were manipulated to have either gestation with lactation or, by removal of pups at birth, gestation without lactation. In addition, hydrocephalus expression in postnatal rats from the breeding colony was analyzed for primiparous and multiparous females. The latter were subdivided according to the interval between the litter examined and the previous litter. There was no particular uterine position or horn that favored hydrocephalus and hydrocephalic fetuses were the same weight as normal littermates. The frequency of hydrocephalus was 16-20% in primiparous females and twofold higher in multiparous females that were lactating during pregnancy. Removal of the suckling pups prevented this increase. The severity of hydrocephalus was measured on 1-mm thick fixed brain slices. Fetuses from lactating females had hydrocephalus that was significantly more severe than the nonlactating groups. However, all fetus groups had hydrocephalus that was very much less severe than the postnatal pups, suggesting that severity increases after birth. It is concluded that there is an epigenetic factor that increases the frequency of inherited hydrocephalus in fetuses if suckling pups are present during gestation. Future experiments will examine possible mechanisms for this genotype-environment interaction. PMID- 12093099 TI - Regeneration of axons after nerve transection repair is enhanced by degradation of chondroitin sulfate proteoglycan. AB - Our past work indicates that growth-inhibiting chondroitin sulfate proteoglycan (CSPG) is abundant in the peripheral nerve sheaths and interstitium. In this study we tested if degradation of CSPG by chondroitinase enhances axonal regeneration through the site of injury after (a) nerve crush and (b) nerve transection and coaptation. Adult rats received the same injury bilaterally to the sciatic nerves and then chondroitinase ABC was injected near the injury site on one side, and the contralateral nerve was injected with vehicle alone. Nerves were examined 2 days after injury in the nerve crush model and 4 days after injury in the nerve transection model. Chondroitinase-dependent neoepitope immunolabeling showed that CSPG was thoroughly degraded around the injury site in the chondroitinase-treated nerves. Axonal regeneration through the injury site and into the distal nerve was assessed by GAP-43 immunolabeling. Axonal regeneration after crush injury was similar in chondroitinase-treated and control nerves. In contrast, axonal regrowth through the coaptation of transected nerves was markedly accelerated and the ingress of axons into the distal segment was increased severalfold in nerves injected with chondroitinase. On the basis of these results we concluded that growth inhibition by CSPG contributes critically to the poor regenerative growth of axons in nerve transection repair. In addition, degradation of CSPG by injection of chondroitinase ABC at the site of nerve repair increased the ingress of axonal sprouts into basal laminae of the distal nerve segment, presumably by enabling more latitude in growth at the interface of coapted nerve. This suggests that chondroitinase application may be used clinically to improve the outcome of primary peripheral nerve repair. PMID- 12093100 TI - Protection of striatal neurons by joint blockade of D1 and D2 receptor subtypes in an in vitro model of cerebral hypoxia. AB - Massive increases in extracellular dopamine have been reported in the ischemic rodent striatum, implicating this neurotransmitter in toxic events. We have examined whether dopamine receptor antagonists are protective against hypoxic insult, using brain slices containing the rostral striatum obtained from adult male C57/BLIcrfa(t) mice. Slices were subjected in vitro to 20 min nitrogen hypoxia, with or without addition of: (i) 50 microM haloperidol (D2 receptor antagonist and sigma ligand), (ii) 10 microM SCH23390 (selective D1 receptor antagonist), (iii) 10 microM eticlopride (selective D2 receptor antagonist), (iv) 10 microM SCH23390 and 10 microM eticlopride in combination, and (v) 10 microM MK 801 (noncompetitive NMDA receptor antagonist). Subsequently, slices were reoxygenated, fixed 2 h postinsult, and processed for light microscopy. Damage was assessed by calculating pyknotic profiles as a percentage of total neuronal profiles present. No pyknotic profiles were detected in normoxic control tissue, but this phenotype predominated in most slices subject to hypoxia alone (60.1 +/- 30.6% pyknotic profiles). Marked protection was produced by haloperidol (7.1 +/- 7.6%, P = 0.002), MK-801 (8.6 +/- 6.9%, P = 0.007), and the combined application of SCH23390 and eticlopride (5.9 +/- 9.4%, P = 0.001). No protection was demonstrated for SCH23390 or eticlopride when applied separately. These data suggest that hypoxic damage in the rostral mouse striatum is mediated via NMDA, D1, and D2 receptors. Protection against hypoxic damage by dopamine receptor antagonists requires the combined blockade of both classes of dopamine receptor. PMID- 12093101 TI - 3-Nitropropionic acid neurotoxicity in hippocampal slice cultures: developmental and regional vulnerability and dependency on glucose. AB - We investigated whether neurotoxic effects of the mitochondrial toxin 3 nitropropionic acid (3-NP) in hippocampal slice cultures are dependent on glucose levels in the culture medium and whether such effects occur via apoptosis or necrosis. In addition, 3-NP toxicity was investigated at two developmental stages of the cultures, prepared from rat brain at postnatal day 5-7 and grown in Neurobasal medium for 1 or 3 weeks. Cultures were exposed to 3-NP in the presence of high (25 mM), normal (5 mM), or low (3 mM) glucose for 48 h, followed by 48 h incubation in medium without 3-NP. Cellular propidium iodide (PI) uptake and lactate dehydrogenase (LDH) efflux into the medium revealed time- and dose dependent cell death by 3-NP, with EC(50) values of about 60 microM in high or normal glucose. Regional vulnerability, as assessed by PI uptake and MAP2 immunostaining, in 3-week-old cultures was as follows: CA1 > CA3 > fascia dentata. In low glucose much lower concentrations of 3-NP (25 microM) triggered neurotoxicity. One-week-old cultures were less susceptible to 3-NP toxicity than 3-week-old cultures, but the dentate granule cells were relatively more affected in the immature cultures. We found no evidence for apoptotic cell death by 3-NP in 3-week-old cultures, but in 1-week-old cultures the putative apoptotic marker c-JUN/AP1 and nuclear fragmentation (Hoechst) were significantly increased in the dentate granule cells. PMID- 12093102 TI - Effect of chronic treatment with riluzole on the nigrostriatal dopaminergic system in weaver mutant mice. AB - The effects of a chronic treatment with the anti-glutamate and sodium channel modulating neuroprotective agent riluzole on the degeneration of dopamine containing neurons were studied in the brain of weaver mutant mice. In these animals, as in Parkinson's disease, dopaminergic neurons of the nigro-striatal pathway undergo spontaneous and progressive cell death. Homozygous weaver mice were orally treated twice a day with either 8 mg/kg riluzole or placebo for 2 months. Quantification of tyrosine-hydroxylase and dopamine-transporter axonal immunostaining in the striatum revealed that riluzole significantly increased the density of striatal dopaminergic nerve terminals. These results suggest that riluzole protects dopaminergic processes in the weaver mice and/or promotes their neuroplasticity. PMID- 12093103 TI - Neurons with mu opioid receptors interact indirectly with enkephalin-containing neurons in the rat dentate gyrus. AB - In the dentate gyrus, mu opioid receptors (MORs) and their enkephalin agonists have overlapping distributions and influence excitability and plasticity. Released endogenous enkephalins can activate at least some of these MORs; however, whether these interactions involve synaptically associated profiles or more distant associations and whether some subcellular compartments (e.g., terminals or dendrites) are more likely to be targeted than others are not known. To elucidate the relationships between potential sites of enkephalin release and MORs, MOR1 and leucine-enkephalin (LE) immunoreactivities were localized in the hilus by electron microscopy, using immunoperoxidase and immunogold markers. Of the 573 MOR-immunoreactive (ir) profiles analyzed, most were axons and terminals (51 and 30%, respectively), and fewer were dendrites (12%), glia (3%), or unclassifiable (4%). Most MOR-ir profiles resembled interneuron processes, while most LE-ir terminals resembled mossy fibers. One third of MOR-ir profiles were within 3 microm and approximately half were within 4 microm of the nearest LE-ir profile. In contrast, few (3%) MOR-ir profiles contacted LE-ir profiles; only 16% of these contacts included observable synapses, and very few profiles (0.5%) colocalized MOR and LE immunoreactivity. MOR-ir axons, terminals, and dendrites were not distributed differently relative to LE-ir profiles. These results suggest that activation of hilar MORs by LE usually involves short-range volume transmission and that dendritic MORs are as likely as axonal and terminal MORs to be activated by released LE. However, the greater abundance of MOR-ir axons and terminals compared to dendrites indicates that presynaptic profiles are a more prominent target for enkephalins and exogenous MOR agonists such as morphine. PMID- 12093105 TI - Olfactory bulbectomy protects hippocampal pyramidal neurons against excitotoxic death. AB - The olfactory system is functionally linked to the hippocampus, and odors can modify the activity of hippocampal neurons. Because hippocampal neurons are selectively vulnerable to death in several prominent neurodegenerative conditions, we tested the hypothesis that activity in olfactory pathways can modify the sensitivity of hippocampal neurons to excitotoxic damage. We report that rats subjected to olfactory bulbectomy exhibit a decrease in the vulnerability of hippocampal pyramidal neurons to excitotoxic injury. Four-month old male Sprague-Dawley rats were subjected to bilateral olfactory bulbectomy or a sham operation. Three months later the rats were given a unilateral infusion of kainic acid in the dorsal hippocampus and were euthanized 24 h later. There was a threefold increase in the number of CA3 neurons that survived kainic acid administration in the bulbectomized rats compared to sham-operated rats. These findings provide the first evidence that olfactory input affects the vulnerability of neurons to excitotoxic death. PMID- 12093104 TI - Mice overexpressing 70-kDa heat shock protein show increased resistance to malonate and 3-nitropropionic acid. AB - Heat shock proteins (HSPs) are induced in response to oxidative stress, hypoxia ischemia, and neuronal injury and play a protective role. Malonate and 3 nitropropionic acid (3-NP) are well-characterized animal models of Huntington's Disease (HD). They inhibit succinate dehydrogenase, inducing mitochondrial dysfunction, which triggers the generation of superoxide radicals, secondary excitotoxicity, and apoptosis. In this study, we examined whether the 70-kDa heat shock protein (HSP-70) is protective against neurotoxicity induced by malonate and 3-NP. Homozygous and heterozygous HSP-70 overexpressing mice (HSP-70+/+, HSP 70+/-) and wild-type controls received 3-NP or malonate and striatal lesion sizes were evaluated by stereology. Compared to HSP-70+/+ and HSP-70+/-, wild-type controls showed significantly larger striatal lesions following 3-NP or malonate injections. These findings support the idea that HSP-70 has a neuroprotective role that may be useful in the treatment of neurodegenerative diseases. PMID- 12093106 TI - Orthographic learning during reading: examining the role of self-teaching. AB - Thirty-four second grade children read target homophonic pseudowords (e.g., slurst/slirst) in the context of real stories in a test of the self-teaching theory of early reading acquisition. The degree of orthographic learning was assessed with three converging tasks: homophonic choice, spelling, and target naming. Each of the tasks indicated that orthographic learning had taken place because processing of target homophones (e.g., yait) was superior to that of their homophonic controls (e.g., yate). Consistent with the self-teaching hypothesis, we obtained a substantial correlation (r=.52) between orthographic learning and the number of target homophones correctly decoded during story reading. Hierarchical multiple regression analyses indicated that neither RAN tasks nor general cognitive ability predicted variance in orthographic learning once the number of target homophones correctly decoded during story reading had been partialed out. In contrast, a measure of orthographic knowledge predicted variance in orthographic learning once the number of targets correctly decoded had been partialed. The development of orthographic knowledge appears to be not entirely parasitic on decoding ability. (c) 2002 Elsevier Science (USA). PMID- 12093107 TI - Experiencing nearby locations together in time: the role of spatiotemporal contiguity in children's memory for location. AB - Three studies investigated how experiencing nearby locations together in time influences memory for location. Seven-, 9-, and 11-year-old children and adults learned 20 object locations in a small-scale space. The space was divided into regions by lines or walls. In Study 1, participants learned the locations either region by region or in a random order. Following learning, participants replaced the objects without the aid of the dots marking the locations and the boundaries subdividing the space. They replaced the objects in any order they chose. After experiencing the locations in random orders during learning, only adults underestimated distances between locations belonging to the same group (i.e., region). Conversely, 9- and 11-year-old children and adults who had experienced the locations region by region during learning underestimated these distances. These findings suggest that experiencing nearby locations together in time increases the weight children assign to categorical information in their estimates of location. Results from Studies 2 and 3 in which participants learned the locations region by region and then replaced the objects region by region (Study 2) or in a random order (Study 3) were similar, highlighting the importance of spatiotemporal cues in memory for location. (c) 2002 Elsevier Science (USA). PMID- 12093108 TI - Perception of speech and nonspeech stimuli by children with and without reading disability and attention deficit hyperactivity disorder. AB - The auditory temporal deficit hypothesis predicts that children with reading disability (RD) will exhibit deficits in the perception of speech and nonspeech acoustic stimuli in discrimination and temporal ordering tasks when the interstimulus interval (ISI) is short. Initial studies testing this hypothesis did not account for the potential presence of attention deficit hyperactivity disorder (ADHD). Temporal order judgment and discrimination tasks were administered to children with (1) RD/no-ADHD (n=38), (2) ADHD (n=29), (3) RD and ADHD (RD/ADHD; n=32), and (4) no impairment (NI; n=43). Contrary to predictions, children with RD showed no specific sensitivity to ISI and performed worse relative to children without RD on speech but not nonspeech tasks. Relationships between perceptual tasks and phonological processing measures were stronger and more consistent for speech than nonspeech stimuli. These results were independent of the presence of ADHD and suggest that children with RD have a deficit in phoneme perception that correlates with reading and phonological processing ability. (c) 2002 Elsevier Science (USA). PMID- 12093109 TI - Developmental patterns in priming and familiarity in explicit recollection. AB - Developmental trajectories of two classes of human memory, implicit and explicit memory, appear to diverge. We examined how developmental differences in perceptual and conceptual priming, two types of implicit memory, coincide with differences between familiarity and recollective responses on explicit memory tests that employ the Remember/Know paradigm ( Tulving, 1985 ). Both types of priming were characterized by developmental invariance in 52 children and adolescents ages 8-19 years. Contrary to Komatsu, Naito, and Fuke (1996) results, few age-group differences in perceptual priming were observed following a levels of-processing encoding manipulation. In contrast, age group differences were found for "Remember" but not "Know" responses. Neither levels of awareness nor strategies influenced priming. Adult levels of performance appear earlier in development on perceptual and conceptual priming tests compared to explicit memory tests. Similar developmental dissociations exist between explicit and implicit memory performance as between "Remember" and "Know" recollective responses. PMID- 12093111 TI - Melatonin, diel rhythms, and metamorphosis in anuran amphibians. AB - Melatonin is present in picogram quantities in the plasma of anuran tadpoles, although the origin of circulating melatonin is not known. Melatonin may have a role in metamorphosis because it is a thyroid antagonist, whose level falls at the metamorphic climax when the thyroid hormones peak. Melatonin rhythms in plasma and eyes are entrained to the light/dark (LD) cycle and affected by temperature. Consequently, melatonin could transduce environmental information to regulate endocrine periodicity and larval circadian organization and influence metamorphic rate. Contradictory results of exogenous melatonin treatment may be largely due to a regulation of the plasma melatonin level which rapidly clears high melatonin concentrations and which can even result in lower circulating melatonin than in controls. Excess exogenous melatonin concentrates in tissues and glands, although the fall in melatonin at metamorphic climax does not occur by this mechanism. There may be thyroxine (T(4))-melatonin interactions at the tissue level that affect metamorphic progress. The rhythms of T(4) and the corticosteroids are also entrained to the LD cycle, and these rhythms, and those of melatonin, change during development, in a specific way on each LD cycle. Differences in the direction and magnitude of phase shifts during development place the peaks of thyroid modulators, such as the corticosteroids and melatonin, in different relationships to the T(4) peaks, which could be an important aspect of the hormonal regulation of metamorphosis. PMID- 12093112 TI - New aspects of signal transduction in the Xenopus laevis melanotrope cell. AB - Light and temperature stimuli act via various brain centers and neurochemical messengers on the pituitary melanotrope cells of Xenopus laevis to control distinct subcellular activities such as the biosynthesis, processing, and release of alpha-melanophore-stimulating hormone (alphaMSH). The melanotrope signal transduction involves the action of a large repertoire of neurotransmitter and neuropeptide receptors and the second messengers cAMP and Ca(2+). Here we briefly review this signaling mechanism and then present new data on two aspects of this process, viz. the presence of a stimulatory beta-adrenergic receptor acting via cAMP and the egress of cAMP from the melanotrope upon a change of alphaMSH release activity. PMID- 12093113 TI - Temperature-dependent prolactin secretion and reproductive biology of the newt Triturus carnifex Laur. AB - The effects of temperature on pituitary prolactin (PRL) gene expression and peripheral levels were studied in both male and female newts obtained from wild conditions during reproductive (Experiment I) and nonreproductive (Experiment II) periods; moreover, changes in parameters related to reproductive function are also described. Male and female newts were taken from a pond in February (Experiment I, reproductive period) and maintained for 1 month in tanks at 4 and 18 degrees C. In male newts kept at 4 degrees C, increase of PRL mRNA in the pituitary and plasma PRL was found compared with that measured in those kept at 18 degrees C. The increase in PRL secretion was parallel to that of plasma androgens and related secondary sexual characteristics (SSC) in males and of plasma estradiol-17beta and vitellogenin in females. On the contrary, in nonreproductive newts (Experiment II), taken from the field in May, no significant changes in plasma PRL, androgens, and SSC were found in those maintained at low temperature (4 degrees C), whereas low temperature significantly increased PRL mRNA expression in the male pituitary and PRL mRNA plus plasma PRL levels in females. These findings suggest that low temperature regulates PRL secretion in this urodele species, showing a sex- and season related control mechanism; moreover, low temperature failed to influence the reproductive biology of newts taken from the field in May, after naturally occurring reproduction during winter months. PMID- 12093114 TI - Cloning, characterization, and comparative activity of turbot IGF-I and IGF-II. AB - IGF peptides belong to a complex system that is known to play a major role in the control of growth and development in mammals. Even if studies performed in nonmammalian species tend to demonstrate an important function of these molecules, use of heterologous ligands, especially in fish, partly limit our knowledge of the physiological role(s) of IGFs. We report in this study the cloning, production, and characterization in an evolved fish, the turbot Psetta maxima, of mature IGF-I and IGF-II. The deduced 68-amino-acid IGF-I and 70-amino acid IGF-II show 75% and 74% sequence identity with their mammalian counterparts, respectively, confirming the high sequence homology observed in other species. The development of a simple and efficient system for the production and purification of both IGF-I and IGF-II in Escherichia coli was used to investigate the in vitro regulation of GH release in the turbot. Our results demonstrated for the first time in a Euteleost species that both peptides specifically inhibited GH release. Both hormones were equally potent in inhibiting GH release from dispersed pituitary cells, with maximal inhibitory effects of 92% and 91% at 1 nM doses after 12 days of culture, respectively. The biologically active recombinant turbot IGFs that we obtained will allow us to further investigate potential and perhaps the specific role(s) of these hormones in turbot as, in contrast with mammals, growth in fish is potentially continued during "adult" life. PMID- 12093115 TI - The cDNA cloning and tissue expression of the cytochrome P450scc from Syrian hamster (Mesocricetus auratus). AB - In this study we report the cloning and sequencing of a cDNA for cholesterol side chain cleavage cytochrome P450scc from Syrian hamster adrenal glands. Isolation of P450scc mRNA was carried out with degenerate primer PCR together with 5' and 3' RACE protocol. The full-length cDNA comprises an open reading frame of 1563 bp encoding a polypeptide of 520 amino acid residues. The predicted protein sequence exhibits well-preserved heme- and steroid-binding domains and shares 89% amino acid sequence identity with rat and mouse enzymes. Transient transfection of HEK 293 cells with the cloned cDNA leads to the formation of pregnenolone from 25 hydroxycholesterol. Northern blot analysis showed expression of mRNAs for P450scc in the major steroidogenic tissues, namely, the adrenal cortex, testis, and ovary. In addition, tissue distribution analysis using the coupled reaction of RT PCR and Southern blotting revealed that the mRNA of the enzyme is also expressed in various nonendocrine tissues, including the epididymis, Harderian gland, and lungs. The relative abundance of specific transcripts at these novel sites suggests that P450scc could potentially play an important role in regulating local steroid hormone synthesis. PMID- 12093116 TI - Effects of the maturation-inducing steroid on LH secretion and the GnRH system at different stages of the gonadal cycle in Atlantic croaker. AB - The effects of treatment with the maturation-inducing steroid 17,20beta,21 trihydroxy-4-pregnen-3-one (20beta-S) on luteinizing hormone-releasing hormone analog (LHRHa)-induced LH secretion were examined during several phases of the gonadal cycle in Atlantic croaker, Micropogonias undulatus. 20beta-S (1 and 5 microg/g of body wt) was administered by intraperitoneal (ip) injection, 24 h prior to injection with LHRHa (10-50 ng/g of body wt) and fish were bled 1 h after LHRHa injection. Treatment with both doses of 20beta-S resulted in plasma concentrations of the steroid within the normal physiological range for this species during final oocyte maturation and ovulation. The 20beta-S treatments altered the LH response to LHRHa throughout the reproductive cycle in both sexes, but the direction and magnitude of the response varied. 20beta-S treatment decreased the LH response to LHRHa in fish with recrudescing and fully recrudesced gonads and in females with regressed gonads. On the other hand, 20beta-S treatment significantly increased the LH response to LHRHa in males with regressing or regressed gonads. 20beta-S treatment also altered preoptic anterior hypothalamic (POAH) and pituitary seabream gonadotropin-releasing hormone (sbGnRH) contents, and the patterns of these changes were similar to those observed in LH secretion. The finding that moderate increases in plasma 20beta-S concentrations, similar to those occurring during final oocyte maturation, significantly inhibit the LH response to LHRHa at the end of the reproductive cycle suggests that this action of 20beta-S is of physiological importance during the periovulatory period. Moreover, the fact that concurrent changes occur in POAH and pituitary sbGnRH contents suggests that the actions of 20beta-S on LH secretion are at least partly mediated via the GnRH system. PMID- 12093118 TI - GH and IGF-I binding in adipose tissue, liver, and skeletal muscle in response to milk intake level in piglets. AB - Two experiments were undertaken to get a better understanding of the regulation of the GH/IGF-I axis by nutritional status in suckling piglets. In experiment 1, 18 newborn unsuckled piglets were bottle-fed low (L), intermediate (I), or high (H) levels of sow colostrum and milk from birth to 7 days of age. L and H levels corresponded to maintenance requirement and ad libitum intake, respectively. Plasma IGF-I and 43-39 kDa IGFBP levels decreased (P<0.05) with the reduction of milk intake. Specific binding of (125)I-bGH in liver decreased (P<0.01) with decreasing milk intake. It did not differ (P>0.1) between the groups in skeletal muscle. Specific binding of (125)I-IGF-I increased (P<0.05) with decreasing milk intake in skeletal muscle but did not differ (P>0.1) between the three groups in liver. In experiment 2, variation in milk intake was induced by litter size manipulation. In the 24 h following birth, the litter sizes of six litters were normalized to 6 (L6) or 12 piglets (L12). Animals were killed at 21 days of age. Plasma IGF-I concentrations (P<0.001) and 43-39 kDa IGFBP levels (P<0.05) were lower in L12 than in L6. Specific binding of (125)I-bGH in adipose tissue, liver, and skeletal muscle did not differ (P>0.1) between the two groups. Specific binding of (125)I-IGF-I in liver was lower (P<0.001) in L12 than in L6, whereas it did not differ (P>0.05) between groups in other tissues. In conclusion, the present report shows that undernutrition consistently decreases plasma IGF-I and 43-39 kDa IGFBP levels but produces variable responses at the GH and IGF-I binding levels in piglets. PMID- 12093117 TI - Increased 21-hydroxylase and shutdown of C(17,20) lyase activities in testicular tissues of the grouper (Epinephelus coioides) during 17alpha-methyltestosterone induced sex inversion. AB - The metabolism in vitro of [(3)H]17-hydroxyprogesterone by gonadal tissues of the grouper (Epinephelus coioides) during 17alpha-methyltestosterone (MT)-induced female-to-male sex inversion was examined. In the female phase, C(17,20) lyase, 5beta-reductase, 3alpha/beta-HSD, 20beta-HSD, and 17beta-HSD activities resulted in the biosynthesis of 5beta-pregnans and 5beta-androstanes (including 5beta androstane-3alpha/beta, 17beta-diol, 3alpha/beta, 17alpha-dihydroxy-5beta-pregnen 20-one, and 5beta-androstane-3,17-dione). In the MT-induced male phase, however, the abrogation of C(17,20) lyase activity and the concomitant activation of 21alpha-hydroxylase/11beta-hydroxylase resulted in the preferential synthesis of polar 21alpha-hydroxlyated 5beta-pregnans (5beta-pregnan 3beta,17alpha,20beta,21alpha-tetrol and 3beta,20beta,21alpha-trihydroxy-5beta pregnan-3-one) and corticosteroids (11-deoxycortisol and cortisol). Interestingly, synthesis of these 21alpha-hydroxylated 5beta-pregnans and corticosteroids was uniquely compartmentalized in only testicular tissues of the MT-induced males. This study shows that there is selective activation of specific steroidogenic enzymes in the different sexual phases leading to the synthesis of metabolites that may be involved in regulating sex inversion of the grouper. PMID- 12093119 TI - In vitro hCG and human recombinant FSH actions on testicular steroidogenesis in the toad Bufo arenarum. AB - In order to study the regulation of testicular steroidogenesis in the toad Bufo arenarum, the effect of gonadotropins (hCG and hrFSH) on steroidogenic enzymes was determined using an in vitro system. 3beta-Hydroxysteroid dehydrogenase/isomerase activity was not affected by any of the gonadotropins, at any of the concentrations used. In contrast, 5alpha-reductase activity was strongly reduced by both hCG and hrFSH. Human chorionic gonadotropin inhibited the activity of cytochrome P450 17alpha-hydroxylase-C(17-20) lyase (P450(c17)), only at the highest concentration used, while hrFSH strongly reduced P450(c17) activity at all the doses assayed. In conclusion, these data suggest that LH (hCG) and FSH regulate steroidogenic enzymes such as 5alphaRed and P450(c17). The results also suggest that FSH could be involved in the regulation of the change in steroidogenesis undergone by the testis during the breeding season. In turn, the inhibition of P450(c17) activity could result in a reduction of androgen production and an increment of C21 steroids. PMID- 12093120 TI - Molecular cloning of three cDNAs encoding different GnRHs in the brain of barfin flounder. AB - To examine the reproductive endocrinology of a large pleuronectiform fish, barfin flounder, Verasper moseri, a promising candidate for aquaculture and resource enhancement in northern Japan due to its high commercial value, three gonadotropin-releasing hormones (GnRHs) in the brain was identified by isolation of their cDNAs. This species had three molecular forms of GnRH; salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II), and seabream GnRH (sbGnRH). Each GnRH cDNA encoded a signal peptide (SP), GnRH, and a GnRH-associated peptide (GAP), which was connected to GnRH by a Gly-Lys-Arg sequence. The sGnRH cDNA encoded an SP composed of 23 amino acids and a GAP composed of 54 amino acids. The cGnRH-II cDNA encoded an SP of 23 amino acids and a GAP of 49 amino acids. The sbGnRH cDNA encoded an SP of 26 amino acids and a GAP of 57 amino acids. In situ hybridization showed that the genes for sGnRH, cGnRH-II, and sbGnRH are expressed in the ventromedial olfactory bulbs and the terminal nerve ganglion, the midbrain tegmentum, and the preoptic area, respectively. These results indicate that sbGnRH neurons in the preoptic area are involved in gonadotropin secretion in barfin flounder. PMID- 12093121 TI - Thyroid hormone-induced protein (TIP) gene expression by 3,5,3(') triiodothyronine in the ovarian follicle of perch (Anabas testudineus, Bloch): modulation of 3beta-hydroxysteroid dehydrogenase/Delta(5)-Delta(4)-isomerase enzyme by TIP. AB - Our previous reports had shown that 3,5,3'-triiodothyronine (T(3)) induced the generation of a 52-kDa monomer protein, i.e., TIP (thyroid hormone-induced protein) in the perch ovarian follicle. TIP, in turn, increased progesterone formation by stimulating Delta(5)-3beta-HSD activity (3beta-hydroxysteroid dehydrogenase/Delta(5)-Delta(4) isomerase) [Eur. J. Endocrinol. 134 (1996) 128 135; Gen. Comp. Endocrinol. 113 (1999) 212-220]. In the present investigation, perch ovarian follicles were incubated in the absence (control) or the presence of T(3) or gonadotropin (GTH) or human chorionic gonadotropin (hCG). RNAs were isolated and allowed to hybridize with a radiolabeled TIP oligonucleotide probe prepared on the basis of the N-terminal 17-amino-acid sequence of TIP. Only RNA from T(3)-incubated follicles hybridized with the probe, while RNA from control or GTH- or hCG-incubated follicles did not hybridize with the probe. The transcript size of TIP mRNA was approximately 1.8 kb. mRNA isolated from T(3) incubated ovarian follicles subjected to in vitro translation and Western blot analysis clearly identified a 52-kDa protein which was not found with the mRNA from the control follicles. However, both TIP and GTH stimulated progesterone secretion from perch ovarian follicles in vitro. GTH stimulation of Delta(5) 3beta-HSD was due to the stimulation of enzyme protein synthesis as a more than twofold increase in Delta(5)-3beta-HSD occurred in response to GTH. But TIP did not stimulate synthesis of Delta(5)-3beta-HSD protein. However, in vitro incubation of Delta(5)-3beta-HSD enzyme with TIP in the presence of NAD and substrate (pregnenolone) greatly stimulated enzyme activity, while incubation with GTH had no effect, indicating a modulation of Delta(5)-3beta-HSD protein from a less active to a more active state by TIP. This has been supported by another observation, in which TIP (52 kDa) and Delta(5)-3beta-HSD (45 kDa) incubation resulted in a complex of 99 kDa. This suggests a protein-protein interaction in the process of Delta(5)-3beta-HSD activation by TIP. The present work, therefore, shows some new and interesting aspects of thyroid hormone regulation of the reproductive control mechanism. PMID- 12093122 TI - Novel biological activities of the fish pro-IGF-I E-peptides: studies on effects of fish pro-IGF-I E-peptide on morphological change, anchorage-dependent cell division, and invasiveness in tumor cells. AB - E-peptides of proinsulin-like growth factor-I (pro-IGF-I) are proteolytically cleaved from the prohormone after translation and have long been regarded as biologically inactive. Tian et al. [Endocrinology 140 (1999) 3387-3390] recently demonstrated that recombinant rainbow trout pro-IGF-I E-peptides (rtEa-2-, rtEa-3 , and rtEa-4-peptides), like hIGF-I, exhibited a dose-dependent mitogenic activity in several nontransformed mammalian cell lines. We show in this report that treatment of established human and fish cancer cells (MCF-7, HT-29, HepG2, ZR-75-1, SK-N-F1, and HC) and retroviral transformed human embryonic kidney cells (293GP) with recombinant rtEa-2- or rtEa-4-, but not rtEa-3-peptide, resulted in a dose-dependent induction of morphological change and enhanced cell attachment. The E-peptide-induced morphological changes are sensitive to treatment with alpha amanitin or cycloheximide, known inhibitors of RNA and protein synthesis. The in vitro colony formation activity of established human tumor cells (HT-29 and MDA MB-231) is greatly reduced or diminished by treatment with the rtEa-4-peptide. Both morphological change and reduction of colony formation activity in MDA-MB 231 cells were also observed following transfection with an Ea-4 transgene construct. Furthermore, the invasive activity of HT1080 cells, known invasive cancer cells, is reduced three to fourfold by treatment with the rtEa-4-peptide. These results suggest that E-peptides of rainbow trout pro-IGF-I possess novel biological activities controlling malignant properties of cancer cells in vitro. PMID- 12093123 TI - Pineal melatonin secretion, but not ocular melatonin secretion, is sufficient to maintain normal immune responses in Japanese quail (Coturnix coturnix japonica). AB - Reports that plasma melatonin is an important immune regulator in avian species have been rather sparse and contradictory. Also, the primary source of immune modulating melatonin has yet to be determined in birds. In Japanese quail (Coturnix coturnix japonica), the pineal gland and eyes contribute roughly two thirds and one third of the melatonin found in the blood, respectively. Two experiments were conducted to evaluate melatonin as an immune modulator in Japanese quail and to determine the primary source of immune-modulating melatonin in this species. Experiment 1 was designed to evaluate the involvement of the pineal gland and the eyes in immunocompetence. Each of three groups of quail was assigned a surgical treatment and the cellular and humoral immune responses were determined 8 weeks following surgery. The surgical treatments were pinealectomy (Px), sham pinealectomy (SH-Px), and ocular enucleation (eye removal (Ex)). Experiment 2 utilized exogenous melatonin as a replacement to reconstitute immune responses in surgically immunocompromised birds. In this experiment, 50.0 microg/ml of melatonin, or diluent only, was provided to Px and SH-Px birds in the drinking water ad libitum. The cellular and humoral immune responses were determined after 8 weeks of melatonin treatment. In both experiments, a cutaneous basophil hypersensitivity reaction to phytohemagglutinin was measured to evaluate the cellular immune response. To evaluate the humoral immune response, primary antibody titers were determined 7 days postintravenous injection with a Chukar red blood cell suspension. Flow cytometric analysis of peripheral blood lymphocytes was performed to determine the relative percentage of CD4(+) and CD8(+) T- and B-lymphocytes in all treatments of Experiment 2. In Experiment 1, both the SH-Px and Ex surgical treatments produced similar cellular and humoral immune responses, and these responses were significantly greater than those in Px treated birds. Pinealectomy significantly reduced the cellular and humoral immune responses from SH-Px by 25.8% and 41.3%, respectively. In Experiment 2, Px again resulted in depressed cellular and humoral immune responses. In addition, Px significantly reduced CD8(+) T-lymphocyte numbers compared to SH-Px, while B lymphocytes remained unchanged. Melatonin administration to Px birds increased the cellular (32.9%) and humoral (30.6%) immune responses to the level of control (SH-Px) birds, although this reconstitution was not due to increased CD8(+) T- or B-lymphocytes. From these data, it was clear that removal of the pineal gland, but not the eyes, reduced cellular and humoral immune responses, which were reconstituted to normal levels by exogenous melatonin. These data suggest that immunodepression is only observed in birds with two thirds of the plasma melatonin removed by pinealectomy. Removal of one third of the plasma melatonin (by ocular enucleation) is not sufficient to reduce cellular and humoral responses in the Japanese quail. PMID- 12093124 TI - Neural transplantation for stroke. AB - Tremendous achievements in neuroscience over the past three decades have provided a solid foundation for basic and clinical research in neurotransplantation. Restorative neurosurgical procedures will develop from different directions, and it is likely that a combination of approaches will be necessary to maximise patient outcomes. We believe that cerebral infarction and selected neurodegenerative disorders are appropriate initial candidates for this research. PMID- 12093125 TI - Does anaesthesia care affect the outcome following craniotomy? PMID- 12093126 TI - The contribution of tremor studies to diagnosis of parkinsonian and essential tremor: a statistical evaluation. AB - This study statistically evaluated a set of commonly measured tremor parameters to determine their individual and combined ability to discriminate between essential tremor (ET) and Parkinsonian tremor (PT). Accelerometer and surface electromyographic (EMG) records of moderate to severe upper limb tremor in 20 patients with ET and 22 patients with PT were used to quantitatively compare tremor amplitude, frequency and pattern of muscle bursting in two resting and three non resting postures. The group statistics showed significant differences between ET and PT with respect to tremor frequency in all five postures, tremor amplitude at rest and muscle bursting patterns. Discriminant function analysis showed that no single parameter or combination of parameters was able to correctly classify all patients. Frequency was much more discriminating than amplitude or muscle bursting patterns in all limb postures. The best amplitude discrimination was obtained when the hand and forearm were both fully supported. Muscle bursting patterns were poorly discriminating and did not assist in correct classification of single patients. Group statistics confirmed a highly significant biological difference between the two tremor types. Optimal classification of single PT (86% correct) and ET (95% correct) patients was obtained using frequency and two selected amplitude parameters from the resting limb. Limb posture was an important variable in optimising the discriminative ability of tremor studies. The implications for routine tremor studies are summarised. PMID- 12093127 TI - Ratings of subjective mental fatigue relate to cognitive performance in patients with myasthenia gravis. AB - We examined ratings of fatigue and cognitive performance in myasthenia gravis (MG) patients and healthy subjects. All participants were administered self report measures of mental and physical fatigue before and after completing a demanding cognitive work battery. Change in fatigue indices was recorded and examined in relation to cognitive function. Results of the study revealed that baseline ratings of fatigue did not relate to cognitive performances for either group. By contrast, increased mental fatigue from the baseline to the post-test assessment correlated with cognitive measures for patients but not control subjects. MG patients reported that physical fatigue also increased following the work battery, but only ratings of mental fatigue correlated with cognitive performances. The results indicate that cognitive impairments may be associated with perceived fatigue in MG. PMID- 12093129 TI - Outcome of children with traumatic brain injury in rural Malaysia. AB - Malaysia had the second highest crude accident rate in the world until 1998. Most children who were involved in these road traffic accidents required intensive neurosurgical care management. We report a prospective study on 36 paediatric neurotrauma patients in rural North East West Malaysia who underwent uniform intensive therapy and were subsequently followed up over a period of 2 years. The modified paediatric Glasgow Coma Scale with support of the revised Wechlser Intelligence Scale for children was used to test the outcome of these children over a period of two years. All patients were managed aggressively in our intensive care as well as our high dependency units. Our results indicate that improvement in outcome is seen after a six month period. Midline shift, duration of coma and duration of transport were found to be significant variables associated with bad outcome. Other variables i.e. age, sex, Glasgow Coma Scale on admission and on site, and lesions of the dominant lobe were not found to be associated with good outcome in these patients. PMID- 12093128 TI - Visual functions in epilepsy patients on valproate monotherapy. AB - Gamma-aminobutyric acid-induced ion transport changes in the retinal pigment epithelium are described. Valproate acts as an inhibitor of gamma-aminobutyric acid transaminase. The purpose of this study was to investigate whether early visual impairment is related to valproate in patients with and without visual symptoms. Thirty-two patients, presenting with a history of seizures currently being treated with valproate, were included in the study. A complete clinical neuroophthalmologic examination was performed, including electroretinogram and visual field test. The electroretinogram parameters of epileptic patients were compared with those of 28 age- and sex-matched healthy volunteers. There was no significant difference in ERG parameters between the two groups. The visual field and visual acuity of all patients were within normal limits. When valproate is not used in conjunction with other antiepileptic drugs and serum levels are within therapeutic levels, it does not cause electrophysiologically detectable retinal dysfunction or any functional defect in visual perception that can be determined clinically. PMID- 12093130 TI - Acute subdural haematomas and enoxaparin. AB - Townsville General Hospital (TGH) has 285 beds and serves a population of approximately 500,0000 in North Queensland. In the period November 1998 until December 1999 there were three acute subdural haematomas (ASDH) related to therapeutic doses of enoxaparin. These culminated in death or severe neurological disability. Unfractionated heparin (UFH) is probably used equivalently in TGH. There were no intracranial haemorrhages in this period with UFH. It is well known low molecular weight heparins (LMWH) have a proven place in the management of deep vein thrombosis (DVT) and ischaemic heart disease (IHD). There may however be some concerns regarding the haemorrhagic complications of therapeutic doses of enoxaparin in the central nervous system. PMID- 12093131 TI - The importance of the intrathecal infusion test in the diagnosis of normal pressure hydrocephalus. AB - The intrathecal infusion test is a reliable method in diagnosing normal pressure hydrocephalus. Between May 1982 and January 2000 we investigated 300 patients suspected of having normal pressure hydrocephalus (NPH) by carrying out an intrathecal infusion test in a constant flow technique. The resistance to cerebrospinal fluid outflow (R(out)) in the intrathecal infusion test was the main criterion of grouping patients into those with normal pressure hydrocephalus or those with cerebral atrophy. A further differentiation into early stage and late stage was made by measuring the compliance (C(p)) - this being the secondary criterion. In 162 patients (54%) the diagnosis of NPH could be confirmed. Of these, 154 patients (95%) underwent a shunt operation. Graduation of NPH and cerebral atrophy following the results of the infusion test at an early stage and an advanced stage allow the conclusion of prognostic evaluations about the course of disease to be made. Patients with NPH in an early stage report in the follow up an improvement of their symptoms after a shunt operation in 65 percent of cases, and those with advanced stage NPH, in 50 percent of cases. The computer aided infusion test allows a reliable differentiation between patients with NPH and those with cerebral atrophy. PMID- 12093132 TI - The morbidity related to atrial fibrillation at a tertiary centre in one year: 9.0% of all strokes are potentially preventable. AB - Atrial fibrillation is a major risk factor for stroke. Anticoagulant therapy reduces this risk but increases the risk of haemorrhage. We aimed to compare the morbidity related to the treatment of atrial fibrillation with warfarin seen in one year at our hospital, with the morbidity in those patients in whom embolism was potentially preventable. There were 111 patients admitted to our hospital in a 12 month period with nonvalvular atrial fibrillation (NVAF) who had stroke, TIA or peripheral embolism. Atrial fibrillation was identified prior to admission in 87 of these 111 (78%) patients with thromboembolism, yet only 14 of these (16%) were receiving warfarin for stroke prophylaxis. Through chart review, a further 56 (64%) patients with embolism could have been receiving anticoagulant therapy if published clinical guidelines(1) were applied. Therefore, 40 episodes of thromboembolism were potentially preventable. Over the same period, there were 18 patients admitted with haemorrhage related to warfarin therapy for stroke prophylaxis in NVAF, including 10 gastrointestinal, five intracerebral, and three peripheral haemorrhages. Most haemorrhages were associated with a high International Normalized Ratio (INR) and the patients were left less disabled than those with embolism. Only one patient with haemorrhage had an absolute contraindication to warfarin therapy (6%). We conclude that the number of preventable strokes far outweighed the morbidity due to warfarin use in the management of NVAF. PMID- 12093133 TI - Cranial venous outflow obstruction in the pseudotumour syndrome: incidence, nature and relevance. AB - This study reports a retrospective analysis of the evidence of cranial venous outflow pathology in 188 patients with pseudotumour syndrome (PTS) investigated over the period 1968-1999. Standard methods of investigation appropriate to the period were used, i.e. cerebral angiography, CT and MR scanning. Recently, some patients had specific venous studies including intraluminal cranial venous sinus pressure measurements. A sub-group (25 patients) was investigated for haematological abnormalities. The overall incidence of cranial venous outflow abnormality was 19.7% (37 cases). In decades, related to the predominant investigative method, the figures were: to 1979, 4.2% (2 cases); to 1989, 15.0% (8 cases); to 1999, 31.0% (27 cases). A cause of the venous abnormality was identified in 20 cases, most commonly haematological and iatrogenic. In 17 patients (all females) no cause was identified. Fifteen of the 25 patients (60%) tested specifically were found to have a haematological abnormality, although no correlation was shown between this and a demonstrable venous outflow abnormality. The conclusion was drawn that there is a high incidence (close to one-third) of venous outflow abnormalities in PTS with detailed investigation. Issues of mechanism and therapy are discussed. PMID- 12093134 TI - Magnesium: a useful adjunct in the prevention of cerebral vasospasm following aneurysmal subarachnoid haemorrhage. AB - Despite recent advances in the management of aneurysmal subarachnoid haemorrhage delayed ischaemic deficits from cerebral vasospasm remains a major cause of morbidity and mortality. As magnesium is a potent cerebral vasodilator we have introduced routine supplementation in patients presented with subarachnoid haemorrhage to determine whether there has been a reduction in the incidence of cerebral vasospasm. METHOD: All patients presented with aneurysmal subarachnoid haemorrhage from February 1997 were included except those who presented after day 5 following bleed. Identical management protocol was used except intravenous magnesium supplementation which was introduced to all patients from May 1999. Incidence of cerebral vasospasm on angiograms among the two groups was analysed. RESULTS: Seven out of 10 patients who did not receive magnesium supplement developed vasospasm requiring intra-arterial papaverine compared with 2 of 13 patients among the treated group (P<0.008). CONCLUSIONS: From our pilot study it appears that magnesium supplement has a beneficial role in the prevention of cerebral vasospasm following aneurysmal subarachnoid haemorrhage. Further studies would seem justified. PMID- 12093135 TI - EGF receptor modifies cellular responses to hyaluronan in glioblastoma cell lines. AB - Cell contact with the extracellular matrix component, hyaluronan, plays a pivotal role in glioma cell invasion and proliferation. Although it is well established that glioma cells can bind hyaluronan to their surface via the expression of CD44, the cellular responses following ligand-receptor interaction remain poorly understood. Given that a large proportion of human high grade gliomas over express the epidermal growth factor receptor (EGFR) and ErbB2, this study aimed to investigate whether an interaction exists between CD44 and these receptor tyrosine kinases. Here we present evidence that CD44 co-immunoprecipitates with EGFR and ErbB2 in the glioma cell lines U87MG and SMA560. Hyaluronan treatment mediated the rapid and transient phosphorylation of extracellular signal regulated kinases 1 and 2 (ERK1 and ERK2) in glioma cell lines. This response to hyaluronan was augmented by the co-expression of EGFR. EGFR also differentially modified the hyaluronan induced expression of a number of genes associated with cellular invasion and proliferation. Northern blot analysis demonstrated that genes encoding urokinase type plasminogen activator (uPA), urokinase type plasminogen activator receptor (uPAR), plasminogen activator inhibitor-1 (PAI-1), tissue inhibitor of metalloproteinases (TIMP-1) and c- myc were up-regulated in response to hyaluronan. Furthermore, zymographic analysis revealed increased levels of uPA in the conditioned medium of hyaluronan stimulated cells. These results indicate a novel functional relationship between CD44 and EGFR in glioma cell lines. The capacity of CD44 to form stable complexes with receptor tyrosine kinases may provide a versatile system for the regulation of cellular invasion and proliferation that allows hyaluronan to activate signal transduction pathways and modulate gene expression via an EGFR-dependent manner. These findings provide new insights into the mode by which hyaluronan regulates the malignant phenotype and also suggest a role for EGFR-CD44 interactions in glial tumorigenesis. PMID- 12093136 TI - Distribution of brain oedema in the contralateral hemisphere after cerebral infarction: repeated MRI measurement in the rat. AB - The appearance of local cerebral dysfunction at remote regions from the focus in the acute stage of stroke (diaschisis) is well known, but its mechanism has not been established. We have analysed serial MR images of the infarcted brain of rats to evaluate the distribution of oedema. Forty-seven Sprague-Dawley rats were anaesthetized with halothane, and the right middle cerebral artery (MCA) was permanently occluded via the intraluminal approach using a nylon 2-0 suture. At 3, 6, 9 and 24 hours after the occlusion, coronal T(2)-weighted MR images were taken and the signal intensity (SI) was computed at each region of the brain. After occlusion of the right MCA, SI increased diachronically up to 24 hours on the occluded side of the cortex (52.9+/-3.2 to 104.8+/-22.4) and striatum, which are within the perfusion territory of the MCA. SI increment was also observed at the hippocampus, alveus hippocampi and pyriform lobe, which are not within the territory of the MCA, and at some regions of the contralateral side (52.5+/-4.8 to 69.4+/-14.8 at the cortex). These changes were prominent in ischaemia vulnerable portions, mild in the cortex, and minimal in the striatum. This contralateral side SI increment indicates remote oedema, which corresponds to diaschisis. We suggest that the mechanism of this remote contralateral oedema is the movement of extravasated protein from the lesion. PMID- 12093137 TI - Frameless stereotaxy as an alternative to fluoroscopy for transsphenoidal surgery: use of the InstaTrak-3000 and a novel headset. AB - Our aim was to evaluate the applicability of an electromagnetic-based frameless stereotactic system for use in transsphenoidal surgery. The system utilizes a novel headset that acts as a replaceable fiducial frame as well as a fixation point for the system's transmitter. It can replace the fluoroscope as a guide to navigation in the sphenoid sinus and sella. The system was used in a consecutive series of 11 patients undergoing transsphenoidal surgery. It was used in conjunction with intraoperative fluoroscopy. We found the Instatrak-3000 to correlate well with lateral fluoroscopic images, and provide additional information in the axial and coronal planes. The InstaTrak-3000 frameless stereotactic system is accurate and obviates the need for intraoperative fluoroscopy during transsphenoidal surgery. Compared to other frameless systems, it avoids the use of fiducial skin markers and head fixation. In common with other frameless stereotactic systems, it provides additional information important for safely approaching the sella and avoids intraoperative radiation exposure for the patient and operating room staff. PMID- 12093138 TI - Patterns of immunocytochemical staining for ferritin and transferrin in the human spinal cord following traumatic injury. AB - Normally Fe(2+) is strictly controlled within the central nervous system (CNS) because of its potential to react with oxygen and form free radicals.(1,2) Traumatic spinal cord injury (TSCI) leads to cell damage and haemorrhage, both of which may increase the pool of free iron.(3) The aim of this study was to examine the response to TSCI of the iron storage protein ferritin (Ft) and the iron transport protein transferrin (Tf). The study found a significant increase in Ft positive cells compared to controls and a significant correlation between the number of Ft positive cells and the severity of injury. Significantly fewer Tf positive cells were seen in the trauma cases compared to the control and there was no relation with the severity of injury. These observations suggest a disturbance in normal iron metabolism within the spinal cord following injury, with possible implications for free radical mediated secondary damage. PMID- 12093139 TI - A technique for safe withdrawal of a catheter tip that hooks a coil loop. AB - In cerebral aneurysm embolization with GDCs, coil movement and subsequent coil protrusion or migration may occur during catheter withdrawal. Coil migration or protrusion usually does not produce clinical problems, but sometimes it may do. Because the consequences of coil movement cannot be anticipated exactly, coil motion during a catheter withdrawal makes interventionists nervous. The authors report on a technique for safe withdrawal of a catheter tip that hooks a coil loop using a small soft coil. PMID- 12093141 TI - Lithium neurotoxicity: the development of irreversible neurological impairment despite standard monitoring of serum lithium levels. AB - We report the case of a 44 year old man who presented with a two-month history of dysarthria, ataxia and leg weakness whilst on maintenance lithium for bipolar disorder. Examination revealed significant cerebellar and pyramidal dysfunction. Serum lithium was 1.5 mmol/l, a moderate elevation above his usual stable levels of 0.4-0.8 mmol/l. The patient's past history included hypertension and chronic renal impairment and the development of neurological symptoms coincided with the recent onset of heart failure. On cessation of lithium he partially recovered, the main residuum being persistent cerebellar ataxia. The case is an example of lithium neurotoxicity developing insidiously in the absence of an overt acute phase syndrome, and highlights the need for keen observation of the patient in the hope of preventing permanent deficits. PMID- 12093140 TI - Magnetic resonance imaging in cases with encephalopathy secondary to immunosuppressive agents. AB - The neurotoxic effects of immunosuppressive agents used after transplantation are well known. In most cases a decrease in drug dosage results in resolution of the neurotoxicity. At early stages in the post-transplantation clinical course, neurotoxicity and other complications such as infectious disease, encephalopathy and seizures are sometimes difficult to diagnose with neuroimaging. Recently, diffusion weighted imaging (DWI) has been used in patients with ischemic disease, mitochondrial myopathy, encephalopathy and demyelinating disease. We examined the magnetic resonance images (MRI), including DWI and fluid attenuated inversion recovery image (FLAIR), in three cases of post-transplantation neurological complication: two cases of neurotoxicity and a case of acute disseminated encephalomyelitis (ADEM). Hyper-intense lesions representing neurotoxicity were seen on FLAIR but not on DWI in two cases with neurotoxicity induced by an immunosuppressive agent. In ADEM, hyper-intense lesions were seen on both FLAIR and DWI. Neurotoxicity due to the immunosuppressive agent showed a favorable outcome, although the hyper-intense lesions temporally presented on FLAIR. In the state after transplantation, hyper-intense lesions on FLAIR and DWI represented in the brain from the initial stage, we might be care of other severe complications but for neurotoxicity. PMID- 12093142 TI - A case of Parkinsonism due to lithium intoxication: treatment with Pramipexole. AB - We report the case of a 72-year-old woman with bipolar disorder treated with lithium for over a year, who suddenly developed disabling Parkinsonism, apparently after a gastroenteric infection. On hospital admission lithium plasma levels were 3.7 mmol/l. After lithium discontinuation, plasma levels decreased to 0.7 mmol/l, but without resolution of neurological symptoms. Biperidene and levodopa-benserazide did not ameliorate her extrapyramidal symptoms. Pramipexole 0.25 mg TID improved dramatically the Parkinsonism and all neurological symptoms resolved in a few days. PMID- 12093143 TI - Ossification of the ligamentum flavum. AB - We report the case of a 32-year-old South Korean male who presented with bilateral leg weakness, spastic gait and associated sensory loss from below the T5 dermatome. MRI and CT scans of the spine confirmed the presence of calcified ligamentum flavum from T1-T6. A thoracic laminectomy resulted in an excellent post-operative recovery with full return of all functions. This case review will discuss the differences that exist between ossification of the posterior longitudinal ligament (OPLL) and ossification of the ligamentum flavum (OLF), but also the similarities at a molecular and possibly at a genetic level. We have reviewed the reported literature of patients presenting with progressive lower limb spacity due to OLF, and an excellent outcome is achieved using decompressive laminectomy. PMID- 12093144 TI - Non-neoplastic demyelinating process mimicking a disseminated malignant brain tumour. AB - Non-neoplastic demyelinating processes of the brain with ring enhancing lesions and mass effect on MRI imaging, mimicking malignant brain tumours, are rare phenomena. We document the case of a 32 year old male with clinical, radiological and initial histological findings, suggestive of a malignant brain tumour. Additional investigations confirmed the diagnosis of multiple sclerosis. This case is significant as the lesion could not be easily distinguished from a malignant brain tumour on imaging alone. Cases such as this illustrate the importance of considering a demyelinating process in the differential diagnosis of tumour-like brain lesions. PMID- 12093145 TI - Spinal schistosomiasis. AB - A 23 year old female presented to Townsville General Hospital seven months after a positive urine test for Schistosomiasis with conus modularize signs. MRI confirmed a conus medullaris enhancing lesion. Serology was positive for Schistosoma IgG : IgM, 3.4 : 1.8. Urine screening for ova, stool sample, rectal biopsy and diagnostic investigations were negative. She was treated empirically for Schistosoma without biopsy with marked resolution of her symptoms and signs. She remains neurologically well one year after presentation. Serological identification from the Centre of Disease Control and Prevention in Atlanta Georgia by western blot has shown positive IgG for Schistosoma haematobium and mansoni. Schistosoma myelopathy is a rare cause of transverse myelitis, conus medullaris syndrome, anterior spinal artery occlusion and radiculopathy in Australia. It should however be included in the differential diagnosis in a patient who has been in or presenting from an endemic area. PMID- 12093146 TI - Intramedullary ancient schwannoma of the cervical spinal cord: case report and review of literature. AB - We report a rare case of intramedullary ancient schwannoma of cervical spinal cord in a 68 year old patient. About 49 cases of intramedullary schwannomas and neurofibromas have been reported in the literature but to our knowledge there is no report of the 'ancient' variety of intramedullary schwannoma. The cell of origin of these tumours is the schwann cell, which normally does not exist in the parenchyma of the central nervous system. Many theories have been advanced to explain this paradox. According to one theory, these tumours arise from the perivascular nerve plexus of the pial vessels. This plexus was found mostly to exist along the branches of the anterior spinal artery. In our case, the tumour was supplied by two branches of the anterior spinal artery, which may add further support to the above theory. PMID- 12093147 TI - Chronic subdural hematoma after transsphenoidal surgery. AB - Chronic subdural haematoma has not been reported as a complication of transsphenoidal surgery. We present a case of pituitary adenoma which was gross totally removed by transsphenoidal surgery with the enlarged sellar opening. Postoperative serial imaging showed massive air accumulation in both frontal subdural spaces followed by subdural fluid collection and subsequent chronic subdural haematomas 2 months later. We conclude that close follow up imaging study is required to avoid overlooking chronic subdural haematoma when massive air is introduced into the subdural space by the transsphenoidal surgery. PMID- 12093148 TI - Paraparesis due to spinal leiomyosarcoma lesion in the thoracic spine accompanied by two leiomyosarcoma lesions in the back and the thigh over an interval of 4 years. AB - We present an 83-year-old man who developed three different leiomyosarcoma lesions in the thigh, back, and thoracic spine. The pathology of the thigh tumour was pleomorphic leiomyosarcoma. MRI after 4 years showed a vertebral bone lesion associated with spinal cord compression at T7 and a paravertebral mass lesion at T9-T10. Surgical treatment for the spinal lesion which caused paraparesis improved the neurological symptoms. The pathological features of the lesions in the back and spine were identical and rather distinct from those of the previous thigh lesion. Spinal leiomyosarcoma causing paraparesis and the two other with soft-tissue leiomyosarcoma lesions with different pathological features in a single patient over a period of 4 years is an extremely rare occurrence. PMID- 12093149 TI - Pathological evidence of encephalomyelitis in the stiff man syndrome with anti GAD antibodies. AB - We report the case of a 57 year old woman with typical clinical features of the stiff man syndrome (SMS) and antibodies to glutamic acid decarboxylase (antiGADAb), who developed a supranuclear gaze palsy, ileus and died of bronchopneumonia eight years after the onset of illness. Post mortem examination revealed perivascular lymphocyte cuffing throughout the cerebral hemispheres, brainstem and spinal cord and neuronal loss in medial anterior horns of the cervical spinal cord. These findings support the notion that the SMS and progressive encephalomyelitis with rigidity (PER) form a clinical and pathologic continuum. PMID- 12093150 TI - Intracranial hypertension in Sturge-Weber/Klippel-Trenaunay-Weber overlap syndrome due to impairment of cerebral venous outflow. AB - Klippel-Trenaunay-Weber syndrome (KTWS) is a rare mesodermal phakomatosis characterized by (1) cutaneous haemangiomata (usually unilateral and involving an extremity) (2) venous varicosities and (3) osseous and soft tissue hypertrophy, also of the affected limb. Sturge-Weber Syndrome (SWS), also a mesodermal phakamatosis, is characterized by meningofacial angiomatosis with cerebral calcification. Overlap between KTWS and SWS is recognized. We describe the case of a young woman with features of both KTWS and SWS who presented with symptoms of acute intracranial hypertension, including headache, vomiting and marked visual impairment. Cerebral angiography revealed paucity of the superficial cortical veins overlying one of the cerebral hemispheres and centripetal venous drainage via small deep venous channels. Based upon this pattern of cerebral venous outflow, we postulate a mechanism to explain the acute episode of intracranial hypertension. PMID- 12093151 TI - On decision-making when a risk is discovered. PMID- 12093152 TI - Munc 18a binding to syntaxin 1A and 1B isoforms defines its localization at the plasma membrane and blocks SNARE assembly in a three-hybrid system assay. AB - Syntaxin 1 and synaptobrevin/VAMP play an essential role in synaptic vesicle exocytosis. Two isoforms for each of these proteins, syntaxins 1A and 1B and synaptobrevin/VAMPs 1 and 2, have been found in nerve endings. Morphological and biochemical studies have revealed a characteristic colocalization and selective interactions patterns of syntaxin 1 and synaptobrevin/VAMP isoforms in nervous and endocrine systems. Moreover, studies in vitro with recombinant proteins have shown characteristic interaction patterns for each syntaxin 1-synaptobrevin/VAMP pair. The cytosolic protein Munc-18a modulates neurotransmission by inhibiting the binding of synaptobrevin/VAMP and SNAP-25 to syntaxin 1A. In the present study, several binding assays were used to demonstrate that Munc-18a significantly binds both isoforms of syntaxin 1 (syntaxins 1A and 1B). Moreover, the coexpression of Munc-18a and syntaxin 1A or syntaxin 1B in 29.3 T cells revealed syntaxin 1-dependent localization of Munc-18a in the plasma membrane. By using the three-hybrid system, we showed the inhibitory role of Munc-18a in the formation of syntaxin 1-synaptobrevin/VAMP complexes regardless of the isoforms. Since Munc-18a can bind both isoforms of syntaxin 1, the present data suggest that this protein is a general modulator of the formation of different SNARE complexes in the nerve endings. PMID- 12093153 TI - Fibroblast growth factor-2 requires glial-cell-line-derived neurotrophic factor for exerting its neuroprotective actions on glutamate-lesioned hippocampal neurons. AB - FGF-2 is a potent neurotrophic factor for several populations of CNS neurons and has been shown to protect hippocampal neurons from glutamate-induced cell death in vitro and in vivo. Mechanisms underlying the neurotrophic and protective actions of FGF-2 have been resolved only in part. Using glutamate-treated cultured hippocampal neurons we show that FGF-2 shares its neuroprotective capacity with GDNF. Hippocampal neurons express glial-cell-line-derived neurotrophic factor (GDNF), its receptors c-Ret and the lipid-anchored GDNF family receptor-alpha1 (GFRalpha-1), and the FGF receptor 1 (FGFR I). Neutralizing antibodies to GDNF abolish the neuroprotective effect of FGF-2. In support of the notion that GDNF is required to permit the protective effects of FGF-2 we find that FGF-2 up-regulates GDNF and GFRalpha-1 in hippocampal neurons. Furthermore, FGF-2-induced GDNF causes enhanced phosphorylation of c-Ret and the signaling components Akt and Erk. A putative downstream target of FGF-2 and GDNF are bcl-2 gene family members, whose mRNAs are differentially up-regulated by the two factors. Together, these data suggest that GDNF is an important protective factor for glutamate-lesioned hippocampal neurons and an essential mediator of the neuroprotective actions of FGF-2. PMID- 12093155 TI - Fatal shift of signal transduction is an integral part of neuronal differentiation: JNKs realize TNFalpha-mediated apoptosis in neuronlike, but not naive, PC12 cells. AB - Tumor necrosis factor-alpha (TNFalpha, 10-100 ng/ml) provokes a dramatic cell death in differentiated PC12 cells (dPC12), but it does not affect the viability and the proliferation of naive PC12 cells (nPC12). We have analyzed the molecular alterations of the TNFalpha-signal cascade underlying this developmental switch toward propagation of apoptosis. The transcriptional inhibitor actinomycin D rendered nPC12 responsive for TNFalpha-induced death, but was hardly effective in dPC12, suggesting that TNFalpha evokes its harmful action in dPC12 predominantly by posttranslational modification of existing molecules. This suggestion was supported by the finding that differentiation of PC12 per se went along with the increased expression of the proapoptotic TNFalpha-receptor I (p55) and its adapter protein Traf-2, whereas expression and phosphorylation of the antiapoptotic Akt (PKB) declined. We could demonstrate that the c-Jun N-terminal kinases (JNKs) mediate this enhanced capacity of apoptotic signaling in dPC12. TNFalpha induced in dPC12, but not nPC12, a biphasic activation of JNKs with a rapid transient JNK1 activation and a second persistent activation of JNK1 and JNK2 paralleled by phosphorylation of c-Jun; in contrast, TNFalpha did not activate p38 kinase. Block of JNKs by CEP-11004, a MLK antagonist and subsequently indirect inhibitor of JNK activation, or L-JNK11, a direct peptidergic inhibitor of JNK activity, almost completely rescued dPC12. Summarizing, the NGF-triggered formation of neurites during differentiation of PC12 includes the reinforced propensity for apoptosis, with JNK2 as the effector in JNK3-negative PC12. These findings offer novel insights into the increased risk of neuronal death, which is linked to the potential to regenerate. PMID- 12093154 TI - Regulation of tau RNA maturation by thyroid hormone is mediated by the neural RNA binding protein musashi-1. AB - The tau gene encodes a microtubule-associated protein expressed by neuronal and glial cells. Abnormal deposits of Tau protein are characteristic of several neurodegenerative disorders. Additionally, mutations affecting tau pre-mRNA alternative splicing of exon 10 are associated with frontotemporal dementia and Parkinsonism linked to chromosome 17. In rodents, this process is developmentally regulated by thyroid hormone (T3) causing the predominance of exon 10-containing transcripts. Here we demonstrate that musashi-1 (msi-1) gene is induced by T3 during rat brain development and in N2a cells. T3 increases msi-1 mRNA level in an actinomycin D-sensitive, cycloheximide-resistant fashion without affecting its half-life, which suggests a transcriptional effect. Both ectopic Msi-1 expression and T3 treatment increased the proportion of exon 10-containing tau transcripts. Furthermore, antisense msi-1 expression inhibited T3 action. Our results show that msi-1 mediates the posttranscriptional regulation of tau expression by T3. PMID- 12093156 TI - Conditionally immortalized clonal cell lines from the mouse olfactory placode differentiate into olfactory receptor neurons. AB - To test extracellular signals that direct the development of the olfactory system, we have generated clonal temperature-sensitive cell lines that represent distinct cellular lineages derived from the E10 mouse olfactory placode. Two of these lines, OP6 and OP27, express (at the permissive temperature), a transcriptional profile representing intermediate-late developmental stages in the olfactory receptor neuron (ORN) lineage. At the nonpermissive temperature, both OP6 and OP27 cells can be induced by all-trans retinoic acid to differentiate into a population of mature bipolar ORN-like cells. In response to retinoic acid, differentiated OP6 and OP27 down-regulate neuron-specific transcription factors required for early stages of neuronal differentiation, and shift active components of the neurotrophin signaling cascade (Trk receptors) into a kinase inactive state. When morphologically mature, OP6 and OP27 express the mature ORN chemosensory signaling components, olfactory G-protein (G(olf)), Type III adenylate cyclase (ACIII), OCNC1, and the olfactory marker protein (OMP). OP27 expresses one odorant receptor, OR 27-3. OP6 expresses two very closely related receptors, OR 6-13 and OR 6-8. Voltage-gated sodium and potassium channels resembling those recorded from primary cultures of ORNs can also be recorded from a subset of differentiated OP6 cells. PMID- 12093157 TI - Acute and chronic acetylcholinesterase inhibition regulates in vivo the localization and abundance of muscarinic receptors m2 and m4 at the cell surface and in the cytoplasm of striatal neurons. AB - Acetylcholinesterase inhibitors (AChE-I) of various pharmacological classes have been used to provoke acute and chronic hypercholinergy in brain. Each condition induces a dramatic decrease of the abundance of muscarinic receptors at the membrane of neurons with simultaneous increase of these receptors in the cytoplasm in association with different subcellular organelles with characteristics depending on the duration of the treatment (short-term versus long term treatment). Each condition also induces a dramatic increase of cytoplasmic receptors associated with endosomes and multivesicular bodies. Chronic treatment with MTF induces a general decrease of m4R in the striatum without modification of the mRNA level but with an exaggerated abundance of muscarinic receptors in the cytoplasm at the sites of synthesis and maturation, i.e., endoplasmic reticulum, nuclear membrane and Golgi apparatus. These results suggest that the membrane abundance and intraneuronal distribution of neurotransmitter receptors are modified following drug treatment with specificity depending on the nature and the duration of treatment. PMID- 12093159 TI - Lysophosphatidic acid influences the morphology and motility of young, postmitotic cortical neurons. AB - Lysophosphatidic acid (LPA) is a bioactive lysophospholipid that produces process retraction and cell rounding through its cognate receptors in neuroblastoma cell lines. Although the expression profile of LPA receptors in developing brains suggests a role for LPA in central nervous system (CNS) development, how LPA influences the morphology of postmitotic CNS neurons remains to be determined. Here we have investigated the effects of exogenous LPA on the morphology of young, postmitotic neurons in primary culture. When treated with LPA, these neurons responded by not only retracting processes but also producing retraction fiber "caps" characterized by fine actin filaments emanating from a dense core. Retraction fiber caps gradually vanished due to the outward spread of regrowing membranes along the fibers, suggesting a role for caps as scaffolds for regrowth of retracted processes. Furthermore, LPA also affects neuronal migration in vitro and in vivo. Taken together, these results implicate LPA as an extracellular lipid signal affecting process outgrowth and migration of early postmitotic neurons during development. PMID- 12093158 TI - Valproate regulates GSK-3-mediated axonal remodeling and synapsin I clustering in developing neurons. AB - Valproate (VPA) and lithium have been used for many years in the treatment of manic depression. However, their mechanisms of action remain poorly understood. Recent studies suggest that lithium and VPA inhibit GSK-3beta, a serine/threonine kinase involved in the insulin and WNT signaling pathways. Inhibition of GSK 3beta by high concentrations of lithium has been shown to mimic WNT-7a signaling by inducing axonal remodeling and clustering of synapsin I in developing neurons. Here we have compared the effect of therapeutic concentrations of lithium and VPA during neuronal maturation. VPA and, to a lesser extent, lithium induce clustering of synapsin I. In addition, lithium and VPA induce similar changes in the morphology of axons by increasing growth cone size, spreading, and branching. More importantly, both mood stabilizers decrease the level of MAP-1B-P, a GSK 3beta-phosphorylated form of MAP-1B in developing neurons, suggesting that therapeutic concentrations of these mood stabilizers inhibit GSK-3beta. In vitro kinase assays show that therapeutic concentrations of VPA do not inhibit GSK 3beta but that therapeutic concentrations of lithium partially inhibit GSK-3beta activity. Our results support the idea that both mood stabilizers inhibit GSK 3beta in developing neurons through different pathways. Lithium directly inhibits GSK-3beta in contrast to VPA, which inhibits GSK-3beta indirectly by an as-yet unknown pathway. These findings may have important implications for the development of new strategies to treat bipolar disorders. PMID- 12093160 TI - Caspr3 and caspr4, two novel members of the caspr family are expressed in the nervous system and interact with PDZ domains. AB - The NCP family of cell-recognition molecules represents a distinct subgroup of the neurexins that includes Caspr and Caspr2, as well as Drosophila Neurexin-IV and axotactin. Here, we report the identification of Caspr3 and Caspr4, two new NCPs expressed in nervous system. Caspr3 was detected along axons in the corpus callosum, spinal cord, basket cells in the cerebellum and in peripheral nerves, as well as in oligodendrocytes. In contrast, expression of Caspr4 was more restricted to specific neuronal subpopulations in the olfactory bulb, hippocampus, deep cerebellar nuclei, and the substantia nigra. Similar to the neurexins, the cytoplasmic tails of Caspr3 and Caspr4 interacted differentially with PDZ domain-containing proteins of the CASK/Lin2-Veli/Lin7-Mint1/Lin10 complex. The structural organization and distinct cellular distribution of Caspr3 and Caspr4 suggest a potential role of these proteins in cell recognition within the nervous system. PMID- 12093161 TI - USP7, a ubiquitin-specific protease, interacts with ataxin-1, the SCA1 gene product. AB - Spinocerebellar ataxia type 1 (SCA1) is an autosomal-dominant neurodegenerative disorder characterized by ataxia and progressive motor deterioration. SCA1 has been known to associate with elongated polyglutamine tract in ataxin-1, the SCA1 gene product. Using the yeast two-hybrid system, we have found that USP7, a ubiquitin-specific protease, binds to ataxin-1. Further experiments with deletion mutants indicated that the C-terminal region of ataxin-1 was essential for the interaction. Liquid beta-galactosidase assay and coimmunoprecipitation experiments revealed that the strength of the interaction between USP7 and ataxin 1 is influenced by the length of the polyglutamine tract in the ataxin-1; weaker interaction was observed in mutant ataxin-1 with longer polyglutamine tract and USP7 was not recruited to the mutant ataxin-1 aggregates in the Purkinje cells of SCA1 transgenic mice. Our results suggest that altered function of the ubiquitin system can be involved in the pathogenesis of spinocerebellar ataxia type 1. PMID- 12093163 TI - Homer-dependent cell surface expression of metabotropic glutamate receptor type 5 in neurons. AB - The metabotropic glutamate (mGlu) receptors are a family of receptors involved in the tuning of fast excitatory synaptic transmission in the brain. Experiments performed in heterologous expression systems suggest that cell surface expression of group I mGlu receptors is controlled by their auxiliary protein, Homer. However, whether or not this also applies to neurons remains controversial. Here we show that in cultured cerebellar granule cells, the group I mGlu receptor subtype, mGlu5, transfected alone is functionally expressed at the surface of these neurons. Transfected Homer1b caused intracellular retention and clustering of this receptor at synaptic sites. Recombinant Homer1a alone did not affect cell surface expression of the receptor, but in neurons transfected with Homer1b, excitation-induced expression of native Homer1a reversed the intracellular retention of mGlu5 receptors, resulting in the receptor trafficking to synaptic membranes. Transfected Homer1a also increased the latency and amplitude of the mGlu5 receptor Ca2+ response. These results indicate that Homer1 proteins regulate synaptic cycling and Ca2+ signaling of mGlu5 receptors, in response to neuronal activity. PMID- 12093162 TI - Sensory neurons respond to hypoxia with NO production associated with mitochondria. AB - Oxygen is pivotal for mammalian cell function, and recent studies suggest an involvement of NO in cellular adaptation to low oxygen supply. Here, we report that endothelial NO-synthase is ubiquitously expressed in rat and mice sensory neurons, and is targeted to juxtamitochondrial compartments of the ER. There it is activated in response to hypoxia while generation of reactive oxygen species remains unaltered. Developing a technique for ultrastructural localization of an NO-sensitive indicator allowed to identify the inner mitochondrial membrane as the target of NO under hypoxia. The demonstrated hypoxic stimulation of endothelial NOS in sensory neurons shall contribute to resistance against hypoxia, since NO promotes cellular survival by interfering with mitochondrial function. PMID- 12093164 TI - Role of heparin-binding growth-associated molecule (HB-GAM) in hippocampal LTP and spatial learning revealed by studies on overexpressing and knockout mice. AB - Heparin-binding growth-associated molecule (HB-GAM) is an extracellular matrix associated protein with neurite outgrowth-promoting activity and which is suggested to be implicated in hippocampal synaptic plasticity. To study the functions of HB-GAM in adult brain we have produced HB-GAM overexpressing mice and compared phenotypic changes in the transgenic mice to those in the HB-GAM null mice. Both mutants were viable and displayed no gross morphological abnormalities. The basal synaptic transmission was normal in the area CA1 of hippocampal slices from the genetically modified mice. However, long-term potentiation (LTP) was attenuated in the mice overexpressing HB-GAM, whereas enhanced LTP was detected in the HB-GAM-deficient mice. Changes in LTP seen in vitro were paralleled by behavioral alterations in vivo. The animals overexpressing HB-GAM displayed faster learning in water maze and decreased anxiety in elevated plus-maze, while the HB-GAM knockouts demonstrated an opposite behavioral phenotype. These results show that HB-GAM suppresses LTP in hippocampus and plays a role in regulation of learning-related behavior. PMID- 12093165 TI - Ocsyn, a novel syntaxin-interacting protein enriched in the subapical region of inner hair cells. AB - Sensory (hair) cells of the inner ear contain two specialized areas of membrane delivery. The first, located at the cell base, is the afferent synapse where rapid delivery of synaptic vesicles is required to convey information about auditory signals with exceedingly high temporal precision. The second area is at the apex. To accommodate the continuous movement of stereocilia and facilitate their repair, recycling of membrane components is required. Intense vesicular traffic is restricted to a narrow band of cytoplasm around the cuticular plate, which anchors stereocilia. Our previous analyses showed that SNARE proteins (syntaxin 1A/SNAP25/VAMP1) are concentrated at both poles of hair cells, consistent with their involvement in membrane delivery at both locations. To investigate further the molecules involved in membrane delivery at these two sites, we constructed a two-hybrid library of the organ of Corti and probed it with syntaxin 1A. Here we report the cloning of a novel syntaxin-binding protein that is concentrated in a previously uncharacterized organelle at the apex of inner hair cells. PMID- 12093167 TI - Rotavirus protein NSP3 shuts off host cell protein synthesis. AB - A recombinant vaccinia virus encoding rotavirus protein NSP3 driven by an internal ribosome entry site (IRES) from the encephalomyocarditis (EMC) virus was able to abate protein synthesis in BSC1 cells by 25-fold, with as much as 30% of the remaining protein synthesis being NSP3. Hence NSP3 shuts off host cell protein synthesis down to the level seen during rotavirus infection but is unable to prevent translation from EMC IRES-driven genes. This effect was abolished by deletions in the eIF4G-binding (aa 274-313) and the dimerization (aa 150-206) but not the viral mRNA-binding (aa 83-149) domains, supporting that NSP3 functions in vivo as a dimer. Binding of eIF4G by NSP3 has been implicated in interfering with mRNA 5'-3' circularization, hence such circularization is essential for translation in mammalian cells. PMID- 12093166 TI - Selective nicotinic receptor consequences in APP(SWE) transgenic mice. AB - The nicotinic (nAChRs) and muscarinic (mAChRs) acetylcholine receptors and acetylcholinesterase (AChE) activity were studied in the brains of APP(SWE) transgenic mice (Tg+) and age-matched nontransgenic controls (Tg-) that were between 4 and 19 months of age. A significant increase in the binding of 125I labeled alpha-bungarotoxin (alpha7 nAChRs) was observed in most brain regions analyzed in 4-month-old Tg+ mice, preceding learning and memory impairments and amyloid-beta (Abeta) pathology. The enhanced alpha7 receptor binding was still detectable at 17-19 months of age. Increase in [3H]cytisine binding (alpha4beta2 nAChRs) was measured at 17-19 months of age in Tg+ mice, at the same age when the animals showed heavy Abeta pathology. No significant changes in [3H]pirenzepine (M1 mAChRs) or [3H]AFDX 384 (M2 mAChRs) binding sites were found at any age studied. The upregulation of the nAChRs probably reflects compensatory mechanisms in response to Abeta burden in the brains of Tg+ mice. PMID- 12093168 TI - Interleukin-8 gene regulation in intestinal epithelial cells infected with rotavirus: role of viral-induced IkappaB kinase activation. AB - Rotavirus is the major etiologic agent of diarrhea in children and the most common cause of severe pediatric gastroenteritis. Rotavirus infection is limited to mature enterocytes that line the villi of the small intestine. Gut epithelial cells, upon infection and cytokine stimulation, are able to produce chemokines, a family of small chemotactic cytokines that regulate the migration and activation of leukocytes. We have previously shown that rotavirus infection of the intestinal epithelial cell line HT-29 induces increased expression of the CXC chemokine interleukin- (IL) 8. Mechanisms responsible for the transcriptional regulation of the IL-8 gene in intestinal epithelial cells during viral infections have not been fully elucidated. Therefore, the purpose of this study was to define the molecular mechanisms of IL-8 gene expression in HT-29 cells infected with rotavirus. Transient transfection analysis of 5' deletions and mutations of the IL-8 promoter driving expression of luciferase reporter gene indicates that the activating protein- (AP) 1 and nuclear factor- (NF) kappaB elements are necessary for IL-8 promoter activation during rotavirus infection. The importance of NF-kappaB activation for IL-8 gene expression was further demonstrated by the inhibition of rotavirus-induced IL-8 gene transcription and protein synthesis following blockade of degradation of the NF-kappaB cytoplasmic inhibitor IkappaB-alpha. Rotavirus infection of HT-29-induced IkappaB kinase (IKK) activation and overexpression of a dominant negative mutant of IKK-beta greatly reduced rotavirus-induced IL-8 promoter activation and NF-kappaB-driven transcription, indicating that IKK is involved in rotavirus-induced IL-8 gene expression and NF-kappaB activation. PMID- 12093169 TI - Interferon regulatory factor-1, interferon-beta, and reovirus-induced myocarditis. AB - Viral myocarditis is an important human disease, and reovirus-induced myocarditis in mice provides an excellent model to study direct viral damage to the heart. Previously, we showed that reovirus induction of and sensitivity to interferon beta (IFN-beta) is an important determinant of viral pathogenicity in the heart and that the transcription factor interferon regulatory factor-3 (IRF-3) is required for reovirus induction of IFN-beta in primary cardiac myocyte cultures. Given several lines of evidence suggesting a possible distinctive environment for IRFs in the heart, we have now focused on IRF-1. Previous studies demonstrated that viruses, double-stranded-RNA (dsRNA), and IFN-alpha/beta can each induce IRF 1 and that IRF-1 plays a role in dsRNA, but perhaps not viral, induction of IFN alpha/beta. Importantly, none of these studies used a virus with a dsRNA genome (such as reovirus), none of them used a highly differentiated nonlymphoid cell type, and none of them addressed whether viral induction of IRF-1 is direct or is mediated through viral induction of IFN-beta. Indeed, as recently as this year it has been assumed that viral induction of IRF-1 is direct. Here, we found that reovirus induced IRF-1 in primary cardiac myocyte cultures, but that IRF-1 was not required for reovirus induction of IFN-beta. Surprisingly, we found that reovirus failed to induce IRF-1 in the absence of the IFN-alpha/beta response. This provides the first evidence that viruses may not induce IRF-1 directly. Finally, nonmyocarditic reovirus strains induced more cardiac lesions in mice deficient for IRF-1 than they did in wildtype mice, directly demonstrating a protective role for IRF-1. Together, the results indicate that while IRF-1 is downstream of the IFN-beta response, it plays an important protective role against viral myocarditis. PMID- 12093170 TI - Hexagonal organization of Moloney murine leukemia virus capsid proteins. AB - To help elucidate the mechanisms by which retrovirus structural proteins associate to form virus particles, we have examined membrane-bound assemblies of Moloney murine leukemia virus (M-MuLV) capsid (CA) proteins. Electron microscopy and image reconstruction techniques showed that CA dimers appear to function as organizational subunits of the cage-like, membrane-bound protein arrays. However, new three-dimensional (3D) data also were consistent with hexagonal (p6) assembly models. The p6 3D reconstructions of membrane-bound M-MuLV CA proteins gave unit cells of a = b = 80.3 A, c = 110 A, gamma = 120 degrees, in which six dimer units formed a cage lattice. Neighbor cage hole-to-hole distances were 45 A, while distances between hexagonal cage holes corresponded to unit cell lengths (80.3 A). The hexagonal model predicts two types of cage holes (trimer and hexamer holes), uses symmetric head-to-head dimer building blocks, and permits the introduction of lattice curvature by conversion of hexamer to pentamer units. The M-MuLV CA lattice is similar to those formed in helical tubes by HIV CA in that hexamer units surround cage holes of 25-30 A, but differs in that M-MuLV hexamer units appear to be CA dimers, whereas HIV CA units appear to be monomers. These results suggest that while general assembly principles apply to different retroviruses, clear assembly distinctions exist between these virus types. PMID- 12093171 TI - Growth characteristics of a highly virulent, a moderately virulent, and an avirulent strain of equine arteritis virus in primary equine endothelial cells are predictive of their virulence to horses. AB - Equine viral arteritis (EVA) is an endotheliotropic viral disease of horses caused by equine arteritis virus (EAV). Although there is only one serotype of EAV, there is marked variation in the virulence of different strains of the virus. The replication and cytopathogenicity of three well-characterized strains of EAV of different virulence to horses were compared in rabbit kidney (RK-13) and primary equine pulmonary artery endothelial cells (ECs). Viral protein expression, plaque size, and cytopathogenicity of all three viruses were similar in RK-13 cells, whereas two virulent strains of EAV were readily distinguished from an avirulent strain by their plaque morphology and cytopathogenicity in primary equine ECs. Furthermore, EAV nucleocapsid protein was detected by flow cytometric analysis significantly later in ECs infected with the avirulent than those infected with the virulent strains of EAV. Primary equine ECs provide a convenient and relevant model for in vitro characterization of the pathogenesis of EVA and the virulence determinants of EAV. PMID- 12093172 TI - Molecular interactions and viral stability revealed by structural analyses of chemically treated cypovirus capsids. AB - Cytoplasmic polyhedrosis virus (CPV, genus Cypovirus) is a unique member of the family Reoviridae which lacks the outer protective shells that exist in all other members, yet exhibits unusual stability. We have analyzed the effects of different acidic, basic, detergent, and urea treatments on CPV capsids. The integrity of the CPV capsids was unaffected under high-pH conditions that disrupted the orthoreovirus inner core, consistent with its ability to maintain structural integrity in extremely alkaline environments during infection. However, it was sensitive to low pH, detergents, and urea, similarly to other viruses in this family. The three-dimensional structure comparisons by electron cryomicroscopy of the intact empty CPV capsid with the "spikeless" capsid whose turrets were removed by chemical treatments revealed the interaction footprint of the turret on the capsid shell. The observed structural changes associated with the removal of the turret suggest critical structural roles of the turret in maintaining capsid integrity in addition to its enzymatic activities. PMID- 12093175 TI - Role of heparan sulfate in human parainfluenza virus type 3 infection. AB - Our current studies have demonstrated that human parainfluenza virus type 3 (HPIV 3) utilizes heparan sulfate (HS) for its efficient cellular entry. HPIV-3 interacted with HS-agarose in vitro and the cellular entry and infection of HPIV 3 were reduced following (a) infection of human epithelial lung A549 cells with HPIV-3 pre-incubated with soluble HS; (b) treatment of A549 cells with heparinase to remove cell surface HS and sodium chlorate (NaClO(3)), a potent inhibitor of proteoglycan sulfation; and (c) infection of HS-deficient mutant CHO cell lines. However, in each instance, complete inhibition of HPIV-3 entry did not occur, suggesting the presence of additional nonproteoglycan cell surface molecule(s) that is required for HPIV-3 entry. Thus the cell surface HS appears to play an important role in efficient cellular entry of HPIV-3. PMID- 12093174 TI - Phylogenetic relationships and differential selection pressures among genotypes of dengue-2 virus. AB - To elucidate the processes controlling the emergence and spread of dengue-2 virus (DEN-2) we examined the evolution of viral isolates sampled from both local (Viet Nam) and global populations. Our phylogenetic analysis, incorporating envelope (E) glycoprotein sequences from 147 isolates of DEN-2, provided a more complete picture of viral diversity, with a newly defined "Cosmopolitan" genotype having a near global distribution and two other genotypes restricted to Asia. By analyzing rates of synonymous and nonsynonymous substitution we determined that genotypes have experienced different selection pressures, with some evidence of positive selection in the Cosmopolitan genotype and one of the two Asian genotypes, but that the transition from sylvatic to human transmission was not accompanied by adaptive evolution of the E gene. Although there was no association between selection pressures acting on the E gene and proposed virulence differences among genotypes, some putatively selected amino acid sites have previously been implicated in changing viral pathogenicity, most notably E-390, and may also affect transmittability. These findings have implications for the future spread of DEN-2. PMID- 12093173 TI - Hepatitis C virus core protein is necessary for the maintenance of immortalized human hepatocytes. AB - Hepatitis C virus (HCV) core protein has many intriguing properties and plays an important role in cell growth regulation. We have recently shown that the HCV core protein from genotype 1a promotes primary human hepatocytes to an immortalized phenotype. Here, we investigated whether the presence of core protein is necessary for maintenance of the immortalized hepatocytes and investigated its consequences on cellular gene expression. The introduction of an antisense orientation of the core gene into immortalized hepatocytes led to the onset of cell death. Further analysis suggested that cell death occurred through apoptosis associated with the activation of tumor suppressor pathways. Antisense core gene expression in immortalized hepatocytes increased p53 expression at both the mRNA and the protein levels. A decreased telomere length and reduced c-myc protein expression were also observed in hepatocytes when the antisense core gene was introduced. Results from these studies suggested that modulation of cell cycle regulatory genes by repression of core protein expression is responsible for reversion of the immortalized phenotype of the hepatocytes. Thus, targeted inhibition may contribute to the development of new therapeutic modalities for prevention of HCV core protein function. PMID- 12093176 TI - Development of feline immunodeficiency virus ORF-A (tat) mutants: in vitro and in vivo characterization. AB - A functional ORF-A is essential for efficient feline immunodeficiency virus replication in lymphocytes. We have characterized a series of mutants of the Petaluma strain, derived from p34TF10 and having different combinations of stop codons and increasingly long deletions in ORF-A. Six clones proved fully replicative in fibroblastoid Crandell feline kidney cells and monocyte-derived macrophage cultures but failed to replicate in T cell lines and primary lymphoblasts. Cats inoculated with three selected mutants had considerably milder infections than controls given intact ORF-A virus. In vivo, the mutants maintained growth properties similar to those in vitro for at least 7 months, except that replication in lymphoid cells was strongly reduced but not ablated. One mutant underwent extensive ORF-A changes without, however, reverting to wild type. Antiviral immune responses were feeble in all cats, suggesting that viral loads were too low to represent a sufficiently powerful antigenic stimulus. PMID- 12093177 TI - Complete genome sequences and phylogenetic analysis of West Nile virus strains isolated from the United States, Europe, and the Middle East. AB - The complete nucleotide sequences of eight West Nile (WN) virus strains (Egypt 1951, Romania 1996-MQ, Italy 1998-equine, New York 1999-equine, MD 2000-crow265, NJ 2000MQ5488, NY 2000-grouse3282, and NY 2000-crow3356) were determined. Phylogenetic trees were constructed from the aligned nucleotide sequences of these eight viruses along with all other previously published complete WN virus genome sequences. The phylogenetic trees revealed the presence of two genetic lineages of WN viruses. Lineage 1 WN viruses have been isolated from the northeastern United States, Europe, Israel, Africa, India, Russia, and Australia. Lineage 2 WN viruses have been isolated only in sub-Saharan Africa and Madagascar. Lineage 1 viruses can be further subdivided into three monophyletic clades. PMID- 12093178 TI - Subcellular localisation, protein interactions, and RNA binding of Potato mop-top virus triple gene block proteins. AB - Subcellular localisation, protein interactions, and RNA binding of the triple gene block proteins (TGBp) of Potato mop-top virus (PMTV) were studied. The 13 kDa (TGBp2) and 21-kDa (TGBp3) proteins with or without green fluorescent protein fused to their N-terminus, and the 51-kDa protein (TGBp1) were expressed individually from a recombinant Tobacco mosaic virus (TMV) vector. Fluorescent images and Western immunoblotting experiments of recombinant TMV-infected Nicotiana benthamiana cells suggested that TGBp2 and TGBp3 were associated with cellular endomembranes and that TGBp3 was associated with the cell wall, possibly located close to plasmodesmata. In Western blots, TGBp1 was detected in fractions containing the cell wall and those enriched for organelles and membranous structures. Self-interactions were demonstrated with all three proteins in yeast two-hybrid experiments, and a heterologous interaction was found between TGBp2 and TGBp3. No additional heterologous interactions were discovered between the different TGBp and none were detected in an in vitro binding assay. TGBp1 and TGBp2 but not TGBp3 were shown to bind ssRNA in a sequence nonspecific manner. The results support the model where TGBp2 and TGBp3 facilitate delivery and localisation of the ribonucleoprotein complex to the plasmodesmata. However, the process is facilitated by RNA-protein rather than protein:protein interactions between the TGBp1 in complex with viral RNA and membrane-localised TGBp2. PMID- 12093179 TI - Binding site analysis of human HBV pol for molecular chaperonin, hsp60. AB - A previous study shows that Hsp60 not only interacts with, but also activates human hepatitis B virus polymerase, HBV Pol (S. G. Park and G. Jung, 2001, J. Virol. 75, 6962-6968). To provide a more detailed analysis of the relationship between the two proteins, (i) the binding sites on human HBV Pol for Hsp60 and (ii) the effect of pregenomic RNA on human HBV Pol-Hsp60 binding were analyzed. The binding sites on human HBV Pol were mapped with several deletion mutant proteins of the Pol expressed in insect cells by using recombinant baculovirus. Immunoprecipitation of each deletion mutant protein by M2 beads showed that binding of Hsp60 to human HBV Pol requires two minimal sites on human HBV Pol: amino acids 1 to 199 (TP) and amino acids 680 to 842 (RH). Human HBV Pol was also shown to bind to Hsp60 in HepG2 cells, the host cell line for human HBV. In addition, Hsp60 binding to the Pol was found to be dispensable to pregenomic RNA binding to human HBV Pol. Overall, this article infers that Hsp60 activates human HBV Pol through binding at the TP and RH domains of the Pol and the Pol binding to Hsp60 does not require pregenomic RNA. PMID- 12093180 TI - Structural features of envelope proteins on hepatitis C virus-like particles as determined by anti-envelope monoclonal antibodies and CD81 binding. AB - The envelope glycoprotein E2 of hepatitis C virus (HCV) is a major component of the viral envelope. Knowledge of its topologic features and antigenic determinants in virions is crucial in understanding the viral binding sites to cellular receptor(s) and the induction of neutralizing antibodies. The lack of a robust cell culture system for virus propagation has hampered the characterization of E2 presented on the virion. Here we report the structural features of hepatitis C virus-like particles (HCV-LPs) of the 1a and 1b genotypes as determined by various mouse and human monoclonal anti-envelope antibodies. Our results show that the E2 protein of HCV-LPs reacts with human monoclonal antibodies recognizing conformational determinants. Monoclonal antibodies (mAbs) specific for the hypervariable region 1 (HVR-1) sequence reacted strongly with HCV-LPs, suggesting that the HVR-1 is exposed on the viral surface. Several mAbs recognized both HCV-LPs with equally high affinity, indicating that the corresponding epitopes [amino acids (aa) 192-217 of E1 and aa 412-423, aa 522 531, and aa 640-653 of E2] are conserved in both genotypes and exposed on the surface of the HCV-LP. The E2 and E1/E2 dimers of 1a bound strongly to the recombinant large extracellular loop (LEL) of CD81 (CD81-LEL) of human and African green monkey, while the HCV-LP of 1a bound weakly to human CD81-LEL. E1/E2 dimers and the HCV-LPs of 1b did not bind CD81-LEL, consistent with the notion that CD81 recognition by E2 is strain-specific and does not correlate with permissiveness of infection. A model of the topology and exposed antigenic determinants of the envelope proteins of HCV is proposed. PMID- 12093181 TI - A novel class of large and infectious defective RNAs of Citrus tristeza virus. AB - Citrus tristeza virus (CTV)-infected plants contain one or more populations of defective RNAs (dRNAs), mostly with a size range of ca. 2.0 to 5.0 kb. Several CTV dRNAs have been characterized and found to consist mainly of the two termini of the genomic RNA, with extensive internal deletions. The present paper describes a new class of large ( approximately 12.0 kb) dRNAs from three different CTV isolates with two unusual features. First is their composition with intact replicase genes. These dRNAs contained a large 5' portion of the genomic RNA terminus, which apparently corresponded to the recently described 5' large single-stranded subgenomic RNA (sgRNA) of ORF1a+1b (Che et al., 2001). The 3' portion of the large dRNAs varied among the 10 different cDNA clones examined in this work. In 2 dRNAs this portion consisted of truncated ORF10 (p20), and in 5 dRNAs it contained truncated ORF11 (p23). Two dRNA molecules were found with a 3' portion that started in the exact 5' position of the intergenic region between the p20 and p23 ORFs. In one dRNA, this portion coincided with the full-length sgRNA corresponding to ORF10. The second unusual feature was their ability to be readily transmitted mechanically to citrus plants by stem slashing and also to Nicotiana benthamiana protoplasts. The possibility that these dRNAs may be encapsidated and be capable of self-replication is discussed. PMID- 12093182 TI - Viremia and immunogenicity in nonhuman primates of a tetravalent yellow fever dengue chimeric vaccine: genetic reconstructions, dose adjustment, and antibody responses against wild-type dengue virus isolates. AB - Chimeric yellow fever (YF)-dengue (DEN) viruses (ChimeriVax-DEN) were reconstructed to correct amino acid substitutions within the envelope genes of original constructs described by Guirakhoo et al. (2001, J. Virol. 75, 7290 7304). Viruses were analyzed and compared to the previous constructs containing mutations in terms of their growth kinetics in Vero cells, neurovirulence in mice, and immunogenicity in monkeys as monovalent or tetravalent formulations. All chimeras grew to high titers [ approximately 7 to 8 log(10), plaque-forming units (PFU)/ml] in Vero cells and were less neurovirulent than YF 17D vaccine in mice. For monkey experiments, the dose of DEN2 chimera was lowered to 3 log(10) PFU in the tetravalent mixture in an effort to reduce its dominant immunogenicity. The magnitude of viremia in ChimeriVax-DEN immunized monkeys was similar to that of YF-VAX, but significantly lower than those induced by wild type DEN viruses. All monkeys developed high levels of neutralizing antibodies against homologous (chimeras) or heterologous (wild-type DEN viruses isolated from different geographical regions) viruses after a single dose of monovalent or tetravalent vaccine. Administration of a second dose of tetravalent vaccine 2 months later increased titers to both homologous and heterologous viruses. A dose adjustment for dengue 2 chimera resulted in a more balanced response against dengue 1, 2, and 3 viruses, but a somewhat higher response against chimeric dengue 4 virus. This indicates that further formulations for dose adjustments need to be tested in monkeys to identify an optimal formulation for humans. PMID- 12093184 TI - New perspectives on bovine tuberculosis. PMID- 12093183 TI - Quasispecies heterogeneity and constraints on the evolution of the 5' noncoding region of hepatitis C virus (HCV): relationship with HCV resistance to interferon alpha therapy. AB - Hepatitis C virus (HCV) polyprotein translation depends on direct internal entry of the 40S ribosomal subunit mediated by an internal ribosome entry segment (IRES) located in the 5' noncoding (5'NC) region of the viral genome. HCV is genetically heterogeneous and is characterized by the existence of a quasispecies distribution of the virus population within a single infected individual. Cloning and sequencing strategies were used to characterize 5'NC quasispecies genetically. Similar to coding regions, the HCV 5'NC region was distributed as a quasispecies, but it appeared to be subjected to stronger conservatory constraints than other regions of the HCV genome, probably due to the need for structural (and functional) conservation of the IRES. Indeed, most of the quasispecies substitutions were in unpaired regions of the IRES or clustered such that base-pairing was maintained, whereas only 21% were expected to result in a loss of base-pairing. Quasispecies-related structural changes could be predicted in the core cruciform of IRES domain III composed of the RNA helices which extend from the four-way junction JIIIabc, mostly in minor variants, but sometimes in major ones. The results presented here suggest the simultaneous presence in infected patients of a mixture of genetically distinct but closely related IRES sequences that may have different structures. No significant genetic changes of 5'NC quasispecies were observed after interferon-alpha treatment, except in patients with mixed genotype infection who cleared one of the infecting strains during therapy, suggesting that the quasispecies distribution of IRES sequences does not play a role in HCV resistance to interferon-alpha therapy. In contrast, the overall quasispecies distribution of HCV genomes (including IRES sequences) might participate in regulation of hepatic and extrahepatic HCV replication. PMID- 12093185 TI - Getting wildlife in perspective - have we thought enough about their diseases? PMID- 12093186 TI - Post-weaning feeding problems in young domestic cats - a new hypothesis. PMID- 12093187 TI - Mycobacterium bovis infection and tuberculosis in cattle. AB - This review considers the possible events that can occur when cattle are exposed to Mycobacterium bovis and, where appropriate, draws on principles accepted for tuberculosis infection in humans and laboratory animal models. Consideration is given to the many complex factors which influence the outcome of challenge with tubercle bacilli. These include features inherent to the mycobacterium, the host and the environment. It is apparent that clinical disease probably occurs only in a relatively small, but undetermined, proportion of cattle that are exposed to Al. bovis. The majority of animals may clear infection or control the bacilli, possibly in a condition of latency. It is concluded that a better understanding of the dynamics of the events following M. bovis exposure and subsequent infection in cattle would be of significant benefit in developing new tools appropriate for disease control and to designing optimal approaches for their application. PMID- 12093188 TI - Wild animals as reservoirs of infectious diseases in the UK. AB - This review aims to illustrate the extent to which wildlife act as reservoirs of infectious agents that cause disease in domestic stock, pet and captive animals and humans. More than 40 agents are described. In the case of some of these, e.g. Cryptosporidium spp., Escherichia coli O157 and malignant catarrhal fever, the current evidence is that wildlife either does not act as a reservoir or is of limited importance. However, in the case of many important diseases, including bovine tuberculosis, Weil's disease, Lyme disease, avian influenza, duck virus enteritis and louping ill, wild animals are considered to be the principal source of infection. Wildlife may be involved in the epidemiology of other major diseases, such as neosporosis, Johne's disease, mucosal disease and foot and mouth disease, but further studies are needed. The UK would benefit from a more positive approach to the study of wildlife and the infections they harbour. PMID- 12093190 TI - Placental transfer of albendazole sulphoxide enantiomers in sheep. AB - Albendazole sulphoxide (ABZSO) is an anthelmintic drug used in veterinary practice. Its molecule has a chiral centre in the sulphur atom and racemic formulations are always used. The kinetics of the ABZSO enantiomers in the last third of pregnancy in ewes, and the placental transfer to the fetus, were studied after a single-dose oral administration (7.5 mg/kg) of a racemic formulation. In mothers, the area under the plasma concentration-time curve (AUC) and C(max) values of (+)-ABZSO (42.4+/-10.5 microg/mL and 1.9+/-0.4 microg/mL, respectively) were higher than those of (-)-ABZSO (15.3+/-5.1 microg/mL and 1.0+/-0.3 microg/mL). The MRT values were 17.0+/-1.6 h for (+)-ABZSO and 13.1+/-1.8 h for ( )-ABZSO. Similar kinetic parameters were obtained in the fetus for both enantiomers, but the fetal concentrations were lower compared with values for the dam. The AUC ratio between (-)-ABZSO/(+)-ABZSO in the dam was 0.36 and in the fetuses 0.64, indicating a higher impairment for the (+)-enantiomer in its placental transfer to the fetus. PMID- 12093189 TI - Coprodiagnosis of Hammondia heydorni in dogs by PCR based amplification of ITS 1 rRNA: differentiation from morphologically indistinguishable oocysts of Neospora caninum. AB - Hammondia heydorni is thought to be a non-pathogenic coccidian parasite of dogs that is closely related to Neospora caninum, an important parasite of cattle and dogs. Oocysts of these two species are morphologically indistinguishable from each other. A population of 2240 dogs in the Czech Republic was screened for the presence of H. heydorni/N. caninum oocysts and five (0.22%), represented by five of 3135 faecal samples (0.16%), were positive. The internal transcribed spacer 1 region of the rRNA gene (ITS1) from two isolates were cloned and the DNA sequences were identical with those of the ITS1 of H. heydorni. Based on the rRNA sequences available for H. heydorni and related coccidia, the primer pair JS4-JS5 was designed to amplify the 3' end of the small subunit (SSU) rRNA gene and ITS1 of H. heydorni. When tested on DNA extracted from a variety of parasites, the primers amplified a specific 267 bp fragment in our isolates only. The presence of DNA equivalent to 10 oocysts was sufficient for the amplification of the ITS1. We present a PCR-based diagnostic method as the only fast and reliable method for the diagnosis of H. heydorni in dogs. PMID- 12093191 TI - Pharmacokinetics of doramectin and ivermectin after oral administration in horses. AB - A study was undertaken in order to compare plasma disposition kinetic parameters of doramectin (DRM) and ivermectin (IVM) in horses after oral administration. Ten crossbreed adult horses, clinically healthy, weighing 380-470 kg body weight (bw) were selected for study. Faecal examinations were performed to determine faecal parasite egg counts. Horses were allocated to two groups of five animals to provide an even distribution considering the variables sex, body weight and faecal egg count. Group I, were treated with an oral paste formulation of IVM at 0.2 mg/kg b/w and Group II, were treated with an oral dose of 0.2 mg/kg bw of DRM prepared as paste from the injectable formulation for oral administration. Blood samples were collected by jugular puncture between 0 h and 75 days post treatment. Plasma was separated and later solid phase extraction and derivatization samples were analysed by high performance liquid chromatography (HPLC); a computerised kinetic analysis was carried out. Data were compared using the Mann-Whitney U-test. The mean plasma concentrations of DRM and IVM after oral administration in horses were detected until 30 and 20 days, respectively. Both drugs showed similar patterns of absorption and no significant differences were found for peak concentration, the time to peak concentration, or for absorptive half-life. The terminal elimination half-life was significantly (P<0.05) longer in the DRM treated group than for the IVM treated group. The differences observed in the elimination half-life explain the longer mean residence time and high values of area under the concentration time curve for the group treated with DRM, which are 30% higher than those of the IVM group. Considering its pharmacokinetics, tolerance and anthelmintic efficacy, the oral administration of DRM, could be an alternative to IVM for the control of parasitic diseases of horses. PMID- 12093192 TI - Clinical findings, bronchoalveolar lavage fluid cytology and matrix metalloproteinase-2 and -9 in canine pulmonary eosinophilia. AB - We characterized clinical and clinicopathological features, and the involvement of gelatinolytic matrix metalloproteinases (MMP-2 and -9) in canine pulmonary eosinophilia (PE). Study material consisted of 20 PE dogs and 16 healthy beagles. All dogs underwent a similar clinical examination and bronchoalveolar lavage (BAL). Analysis for cell count and differential cell count of BAL fluid (BALF), arterial blood gas analysis before and after BAL, and thoracic radiographs before BAL and after treatment were obtained. Twelve dogs were re-evaluated and six relavaged. MMP-2 and MMP-9 in BALF were analysed by zymography, Western immunoblotting and immunocytochemistry. In the PE dogs, BALF, cell count, number and percentage of eosinophils, and numbers of macrophages, lymphocytes, neutrophils, mast cells and epithelial cells were all significantly elevated. Blood eosinophilia was detected in half of the PE dogs. Three PE dogs had mild hypoxaemia. The BAL procedure had an equal effect on PE and healthy dogs' arterial blood gas values. Bronchointerstitial densities were seen in PE dogs' radiographs. Treatment of PE decreased BALF cell count, eosinophil count and percentage and diminished radiographic changes. Gelatinolytic activity was higher in PE dogs' BALF. BALF macrophages and epithelial cells were the principal sources of the MMP-9. PMID- 12093193 TI - The absorption of phosphate ions from the ovine reticulorumen. AB - The reticulorumen is now recognised to be an important site of net absorption of phosphate ions from ruminal fluid containing phosphate concentrations appropriate to those found in normal farming practice. These rates of absorption were measured in vivo from solutions placed in the washed reticulorumen, isolated in situ, in conscious, trained sheep. Reducing the ruminal sodium concentration led to reduced absorption of phosphate, suggestive that phosphate and sodium fluxes across the apical wall of the ruminal epithelial cell are linked, as they are in the kidney. Increased absorption of short chain fatty acids led to enhanced absorption of phosphate ions. Conversely, inhibition of carbonic anhydrase activity, by the addition of 1 mM acetazolamide to the ruminal fluid, led to a reduction in phosphate absorption. An increase in the acidity of the ruminal fluid also increased the absorption of phosphate, as did an increase in the ruminal Ca(2+) concentration over the range 1-4 mmol per litre. It is suggested that these effects can be accounted for by a Na(+)/H(+) antiporter coupled with a phosphate/proton symporter in the apical membrane of the ruminal epithelial cell. PMID- 12093194 TI - Relationship between clinical signs and pulmonary function estimated by the single breath diagram for CO(2) (SBD-CO(2)) in horses with chronic obstructive pulmonary disease. AB - The pulmonary health of 66 horses was assessed by a clinical examination and simple supplementary diagnostic methods. Single breath diagrams for CO(2) (SBD CO(2)) and derived lung function indices were used to determine pulmonary function. The clinical signs in different groups were related to the results of the lung function indices derived from the SBD-CO(2). In horses with moderate to severe chronic obstructive pulmonary disease (COPD), a significant relationship was found between the respiratory frequency and the ratio of Bohr's dead space to the tidal volume (VD(Bohr)/VT), and between the physiological dead space/tidal volume ratio (VD(phys)/VT) and the ratio of the alveolar dead space to the alveolar tidal volume (VD(alv)/VT(alv)), but no significant associations were found between the arterial oxygen tension (P(a)O(2)) and lung function indices derived from the SBD-CO(2). The occurrence of cough, the viscosity of tracheobronchial mucus and the amount of polynuclear neutrophils in tracheobronchial aspirates were significantly related to the expiratory tidal volume (VT), the total expired volume of CO(2) (VCO(2)), VD(Bohr)/VT, VD(phys)/VT and VD(alv)/VT(alv). We conclude that abnormal findings in these clinical parameters indicate a measurable ventilation and perfusion (V(A)/Q) mismatch which is reflected by increases in dead space, VD(Bohr)/VT and VD(phys)/VT as well as VD(alv)/VT(alv). PMID- 12093196 TI - A retrospective study of oesophageal endoscopy in cattle - oesophagoscopy for diagnosis of mucosal disease. AB - Oesophageal endoscopy in cattle has been rarely reported in the literature. It has, however, been used for many years as an aid to the diagnosis and treatment of illnesses of the upper gastrointestinal tract in small animals and horses. We have assessed the potential value of oesophagoscopy on 120 bovine patients presented at the 2nd Medical Clinic for Ruminants and Swine at the University of Veterinary Medicine, Vienna, over a period of three years. In the course of the study, cattle exhibiting symptoms such as salivation, regurgitation or swelling in the neck region were referred for endoscopic examination. The main indication was to confirm or exclude bovine virus diarrhoea virus infection in calves, in which typical erosions were visible endoscopically on the mucosa of the oesophagus in infected animals. In addition, endoscopy proved to be useful for the diagnosis of diverticula, ruptures and inflammation of the oesophagus. Oesophagoscopy can be considered to be a valuable supplementary aid to clinical examination and, in many cases, can facilitate diagnosis. PMID- 12093195 TI - Biomechanical and histopathological changes in the support structures of bovine hooves around the time of first calving. AB - Hooves were removed from heifers killed two weeks before calving (C-2), and four and twelve weeks post-calving (C+4 and C+12), and from age-matched maiden heifers. Segments were dissected from the anterior walls of lateral hind claws, to include horn, corium and bone, and the biomechanical properties were measured. There was reduced rigidity, both overall and at a physiologically appropriate displacement of 2 mm, and a greater displacement was required to reach initial and maximum support in the hooves of calving, compared with maiden, heifers. Histological examination revealed widened and distorted interdigitating laminae, progressing to C+12 in the hind claws, but with evidence of recovery in the front claws after C+4. Increased connective tissue strength from dorsal to ventral hoof segments was correlated with protein, proteoglycan, pro- and activated matrix metalloproteinase-2, and tissue inhibitors of metalloproteinases, and was inversely correlated with fat, water and collagen content. This implies that mechanical changes reflect alterations in the biochemistry of the connective tissue. This evidence supports the hypothesis that primary causal events associated with calving weaken the connective tissue of the hoof suspensory apparatus, leading to increased susceptibility to clinical lameness associated with sole ulcers and white line disease. PMID- 12093197 TI - Serum and synovial fluid concentrations of bovine chondrocalcin in dogs. PMID- 12093198 TI - Lambs with white muscle disease and selenium content of soil and meadow hay in the region of Kars, Turkey. PMID- 12093199 TI - Septicaemic pasteurellosis in ostriches (Struthio camelus) in central Saudi Arabia. PMID- 12093200 TI - An early invasive strategy was effective in the acute coronary syndrome with elevated cardiac troponin I and T levels. PMID- 12093201 TI - Review: glycoprotein IIb/IIIa inhibitors reduced death or MI in acute coronary syndromes not routinely scheduled for revascularization. PMID- 12093202 TI - Review: interventions that lower cholesterol levels or blood pressure in diabetic patients prevent cardiovascular disease. PMID- 12093203 TI - Treating isolated systolic hypertension prevented major cardiovascular events across strata of risk in older patients. PMID- 12093204 TI - Review: antiplatelet therapy prevents occlusive vascular events in high-risk patients. PMID- 12093205 TI - Review: aspirin reduces CAD events in persons with no history of cardiovascular disease, but it increases gastrointestinal bleeding. PMID- 12093207 TI - Cardiac pacing reduced nonaccidental falls in older adults with cardioinhibitory carotid sinus hypersensitivity. PMID- 12093208 TI - Review: intrinsic and environmental risk-factor modification reduces falls in elderly persons. PMID- 12093206 TI - A preoperative smoking intervention decreased postoperative complications in elective knee or hip replacement. PMID- 12093209 TI - Review: selegiline leads to a small short-term improvement in cognition and activities of daily living in Alzheimer disease. PMID- 12093211 TI - Abciximab added to urokinase increased amputation-free survival in peripheral arterial occlusion of the legs. PMID- 12093210 TI - Review: extended out-of-hospital prophylaxis with heparin prevents deep venous thrombosis in elective hip arthroplasty. PMID- 12093212 TI - Helicobacter pylori eradication reduced the risk for ulcers in dyspeptic patients who were about to begin NSAID treatment. PMID- 12093213 TI - Adding intranasal fluticasone to cefuroxime resolved acute rhinosinusitis. PMID- 12093214 TI - Review: oral and intravaginal agents are equally effective for treatment of uncomplicated vulvovaginal candidiasis. PMID- 12093215 TI - Verbal advice plus an information leaflet reduced antibiotic use in acute bronchitis. PMID- 12093216 TI - Review: interferon-alpha plus ribavirin improves the virologic response in chronic hepatitis C. PMID- 12093217 TI - The inactivated trivalent split-virus influenza vaccine was safe in stable asthma. PMID- 12093218 TI - Low-dose budesonide improved asthma control in mild asthma; adding formoterol improved control in corticosteroid-treated patients. PMID- 12093219 TI - Montelukast improved pulmonary function and asthma-specific quality of life in aspirin-intolerant asthma. PMID- 12093220 TI - Sildenafil improved erectile dysfunction and quality of life in men with comorbid mild-to-moderate depression. PMID- 12093221 TI - A multidisciplinary community-based rehabilitation program improved social functioning in severe traumatic brain injury. PMID- 12093222 TI - Review: hospital-at-home care for early discharge or admission avoidance does not improve health outcomes. PMID- 12093223 TI - Review: bed rest does not prevent cervical or lumbar puncture headaches. PMID- 12093224 TI - Review: St. John's wort, ginkgo, saw palmetto, and kava may be effective for some conditions. PMID- 12093225 TI - Rofecoxib, 25 mg/d, was more effective than rofecoxib, 12.5 mg/d, celecoxib, or acetaminophen in osteoarthritis of the knee. PMID- 12093226 TI - Naltrexone for 3 or 12 months did not reduce drinking in alcohol dependence. PMID- 12093227 TI - Review: behavioral interventions plus laxatives are effective for defecation disorders, but biofeedback does not add benefit. PMID- 12093228 TI - Review: a home-like birth environment has beneficial effects on labor and delivery. PMID- 12093229 TI - Review: antibiotics improve maternal and fetal outcomes and are safe in preterm, prelabor rupture of membranes. PMID- 12093230 TI - Oral dexamethasone led to fewer treatment failures than did nebulized dexamethasone or placebo in children with mild croup. PMID- 12093232 TI - Magnetic resonance direct thrombus imaging was accurate in suspected deep venous thrombosis. PMID- 12093231 TI - Growth rate was greater with fluticasone propionate than with beclomethasone dipropionate in children with chronic asthma. PMID- 12093233 TI - Review: interventions for health care providers improve provider-patient interactions and patient satisfaction. PMID- 12093234 TI - A comprehensive annotated reminder tool increased appropriate screening in primary care. PMID- 12093235 TI - A risk index with 14 variables predicted 30-day postoperative pneumonia after major noncardiac surgery. PMID- 12093236 TI - A simple scoring system predicted clinical progression in HIV patients receiving highly active antiretroviral therapy. PMID- 12093237 TI - Textbook descriptions of disease--where's the beef? PMID- 12093238 TI - Revascularization led to less angina and fewer adverse cardiac events than did optimal medical care in angina pectoris in the elderly. PMID- 12093239 TI - Updated definitions of healthy ranges for serum alanine aminotransferase levels. AB - BACKGROUND: Serum alanine aminotransferase (ALT) activity, the variable most commonly measured to assess hepatic disease, fails to identify many patients with hepatic injury. Current standards for "normal" ALT level were defined by using populations that included persons with subclinical liver disease. OBJECTIVE: To update definitions of healthy ranges for serum ALT level. DESIGN: Retrospective cohort study. SETTING: A university hospital in Milan, Italy. PARTICIPANTS: 6835 persons who were first-time blood donors from 1995 through 1999, were negative for anti-hepatitis C virus (HCV), and had no contraindications to donation and 209 persons who attempted to donate blood from 1990 through 1999 but were found to have anti-HCV antibodies. Of the latter group, 131 had HCV viremia. MEASUREMENTS: Univariate and multivariate analyses examined associations between clinical and laboratory factors and ALT levels. Healthy ranges for ALT were computed from the population at lowest risk for liver disease. Sensitivity and specificity of healthy ALT ranges were evaluated in the donors with HCV antibodies, of whom 133 had liver biopsy. RESULTS: Serum ALT activity was independently related to body mass index and to laboratory indicators of abnormal lipid or carbohydrate metabolism. Updated upper limits (for men, 500 nkat/L [30 U/L]; for women, 317 nkat/L [19 U/L]) were lower than current limits (for men, 667 nkat/L [40 U/L]; for women, 500 nkat/L [30 U/L]) and, during 6-month follow up, showed superior sensitivity in identifying participants with HCV viremia (sensitivity, 76.3% [95 % CI, 69.1% to 83.6%] vs. 55% [CI, 46.4% to 63.5%]). The related tradeoff in specificity was acceptable (88.5% [CI, 79.2% to 94.6%] vs. 97.4% [91% to 99.7%]). The increased sensitivity targeted patients with minimal to mild histologic lesions. CONCLUSION: In patients with chronic HCV infection or nonalcoholic fatty liver disease, revision of normal limits for ALT level is advisable. PMID- 12093240 TI - Reporting the recruitment process in clinical trials: who are these patients and how did they get there? AB - BACKGROUND: A common criticism of randomized, controlled trials (RCTs) is that the enrollment process may be highly selective and those who enroll may not represent persons in the general population. The recruitment process reported in published RCTs has not been systematically evaluated. OBJECTIVE: To determine whether published RCTs report information about how their study sample was assembled and to describe the proportion of potential study participants who were actually enrolled. DESIGN: Cross-sectional explicit review of RCTs published in four high-impact medical journals between 1 April 1999 and 1 April 2000. All RCTs involved interventions in humans. MEASUREMENTS: The number of persons who were screened for eligibility, the number who were eligible, and the number who were enrolled in each RCT. RESULTS: A total of 172 RCTs were reviewed. Ninety (52%) reported the number of persons who were evaluated by the investigators for eligibility, and 74 (43%) reported the number of persons who were actually eligible for participation. Of the studies that reported quantitative recruitment information, the median proportion of screened persons who were eligible for participation was 65% (interquartile range, 41% to 82%) and the median proportion of eligible persons who enrolled was 93% (interquartile range, 79% to 100%). Some trials reportedly enrolled every person screened for eligibility; others screened as many as 68 people for each person finally enrolled. CONCLUSIONS: Many RCTs published in major medical journals do not provide information about the patient recruitment process. As a result, it is difficult for readers to gauge the extent to which participants may represent a highly selected subgroup. PMID- 12093241 TI - High prevalence of osteonecrosis of the femoral head in HIV-infected adults. AB - BACKGROUND: Osteonecrosis has been reported to occur occasionally among HIV infected patients. The diagnosis of symptomatic osteonecrosis of the hip in two of the authors' patients, together with reports from community physicians, raised a concern that the prevalence of osteonecrosis is increasing. OBJECTIVE: To determine the prevalence of osteonecrosis of the hip in asymptomatic HIV-infected patients and to identify potential risk factors associated with osteonecrosis. DESIGN: Survey and comparison study. SETTING: The Clinical Center of the U.S. National Institutes of Health. PARTICIPANTS: 339 asymptomatic HIV-infected adults (of 364 asked to participate) and 118 age- and sex-matched HIV-negative volunteers enrolled between 1 June and 15 December 1999. MEASUREMENTS: Osteonecrosis of the hip, as documented by magnetic resonance imaging. Data from clinic records and a patient questionnaire administered before magnetic resonance imaging were used in an analysis of risk factors. A subset of patients was evaluated for hypercoagulable state. RESULTS: Fifteen (4.4% [95% CI, 2.5% to 7.2%]) of 339 HIV-infected participants had osteonecrosis lesions on magnetic resonance imaging, and no HIV-negative participants had similar lesions. Among HIV-infected participants, osteonecrosis occurred more frequently in those who used systemic corticosteroids, lipid-lowering agents, or testosterone; those who exercised routinely by bodybuilding; and those who had detectable levels of anticardiolipin antibodies. CONCLUSIONS: Patients infected with HIV have an unexpectedly high occurrence of osteonecrosis of the hip. Although screening asymptomatic patients is not warranted, HIV-infected patients with persistent groin or hip pain should be evaluated for this debilitating complication. PMID- 12093242 TI - Metformin: an update. AB - Metformin is an insulin-sensitizing agent with potent antihyperglycemic properties. Its efficacy in reducing hyperglycemia in type 2 diabetes mellitus is similar to that of sulfonylureas, thiazolidinediones, and insulin. Metformin based combination therapy is often superior to therapy with a single hypoglycemic agent. The antihyperglycemic properties of metformin are mainly attributed to suppressed hepatic glucose production, especially hepatic gluconeogenesis, and increased peripheral tissue insulin sensitivity. Although the precise mechanism of hypoglycemic action of metformin remains unclear, it probably interrupts mitochondrial oxidative processes in the liver and corrects abnormalities of intracellular calcium metabolism in insulin-sensitive tissues (liver, skeletal muscle, and adipocytes) and cardiovascular tissue. PMID- 12093243 TI - Annals of Internal Medicine at age 75: reflections on the past 25 years. PMID- 12093244 TI - Reflections on medical journals: has progress made them better? PMID- 12093245 TI - Alanine aminotransferase levels: what's normal? PMID- 12093247 TI - Making good decisions about breast cancer chemoprevention. PMID- 12093249 TI - Chemoprevention of breast cancer: recommendations and rationale. PMID- 12093250 TI - Chemoprevention of breast cancer: a summary of the evidence for the U.S. Preventive Services Task Force. AB - PURPOSE: Chemoprevention offers promise as a strategy for reducing morbidity and mortality from breast cancer in women. This review examined the evidence for the effectiveness of chemoprevention in women without a history of breast cancer. DATA SOURCES: MEDLINE (1966 to December 2001). STUDY SELECTION: English-language, randomized, controlled trials (RCTs) of chemoprevention of breast cancer in women without a previous diagnosis of breast cancer were examined, and 4 relevant trials, 3 involving tamoxifen and 1 involving raloxifene, were selected. Trials that provided data on the harms of tamoxifen or raloxifene, studies of the costs of chemoprevention, and studies of risk assessment were also reviewed. DATA EXTRACTION: Four reviewers independently abstracted data on key variables, including study population, sample size, randomization, treatment, and outcomes. DATA SYNTHESIS: The largest of the RCTs of tamoxifen reported a 49% reduction in relative risk (0.51 [95% CI, 0.39 to 0.66]) for invasive cancer among women with an estimated 5-year breast cancer risk of at least 1.66%. The other tamoxifen trials did not observe a statistically significant benefit, but only a few women in each trial took tamoxifen during the entire study period. The raloxifene study of postmenopausal women with osteoporosis found a 76% reduction in relative risk (0.24 [CI, 0.13 to 0.44]) for invasive breast cancer. Tamoxifen and raloxifene were effective only against estrogen receptor-positive tumors. Both drugs increased risk for venous thromboembolic disease and hot flashes; tamoxifen increased risk for endometrial cancer and stroke. CONCLUSIONS: Tamoxifen and raloxifene reduce the incidence of estrogen receptor-positive breast cancer in women. The relative risk reduction seems similar across all breast cancer risk groups. The absolute risk reduction varies by risk factors for breast cancer, however, and must be balanced against the potential harms to judge the appropriateness of treatment for individual women. PMID- 12093251 TI - Sinusitis. PMID- 12093252 TI - Quality of care in patients with diabetes. PMID- 12093253 TI - Quality of care in patients with diabetes. PMID- 12093254 TI - Quality of care in patients with diabetes. PMID- 12093258 TI - Measuring patient adherence. PMID- 12093259 TI - Spontaneous intramural hematoma of the esophagus. PMID- 12093260 TI - Severe rhabdomyolysis related to cerivastatin without gemfibrozil. PMID- 12093261 TI - Gabapentin toxicity requiring intubation in a patient receiving long-term hemodialysis. PMID- 12093262 TI - Celecoxib-induced cholestatic hepatotoxicity in a patient with cirrhosis. PMID- 12093263 TI - Summaries for patients. New definitions for healthy ranges of alanine aminotransferase, a blood test of liver function. PMID- 12093264 TI - Summaries for patients. Reporting the recruitment process in clinical trials: who are these patients and how did they get there? PMID- 12093265 TI - Summaries for patients. Problems of the hip bone in patients with HIV infection. PMID- 12093266 TI - Summaries for patients. Metformin for patients with type 2 diabetes mellitus. PMID- 12093267 TI - Summaries for patients. Using medication to prevent breast cancer: recommendations from the United States Preventive Services Task Force. PMID- 12093268 TI - Adenosine and Aggrenox: a hazardous combination. PMID- 12093269 TI - Absolute measurement of phosphorylation levels in a biological membrane using atomic force microscopy: the creation of phosphorylation maps. AB - We show that it is possible to produce phosphorylation difference maps of biological membranes under conditions which reflect those in vivo and in which proteins remain functional. We also demonstrate that absolute levels of phosphorylation are retrieved through the application of an appropriate calibration method. Finally we show that the kinetics of phosphorylation/dephosphorylation can also be monitored. These methods are demonstrated on photosynthetic membranes from higher plants, for which protein phosphorylation is the dominant regulatory mechanism. We show directly that the most recent estimates of the phosphorylation levels in this system are reasonably accurate. Phosphorylation difference maps show that the distribution of phosphates is not even, with significantly higher levels at the membrane margins and patches of high phosphate density next to patches of low charge density. PMID- 12093270 TI - Interaction between the antiapoptotic protein Nr-13 and cytochrome c. Antagonistic effect of the BH3 domain of Bax. AB - Mitochondria act as a focal point for upstream apoptosis signals by releasing cytochrome c into the cytosol, leading to the activation of caspases and subsequent cell death. Members of the Bcl-2 protein family regulate this phenomenon by heterodimerization via the BH3 domain of proapoptotic members opposing their pro- and antiapoptotic functions. The mechanism of cytochrome c release from mitochondria and of its regulation remains controversial. In vitro binding studies of purified and biologically active proteins should help in understanding the molecular mechanism of interactions and protein functions. In this work, the Bcl-2-related antiapoptotic chicken protein Nr-13 was overexpressed as a highly soluble recombinant protein which showed correct folding as judged by circular dichroism and fluorescence spectroscopy. Purified Nr-13 inhibits caspase-3 activation in a Xenopus egg-derived cell-free system, and neutralizes the proapoptotic activity of a synthetic peptide containing the BH3 domain of Bax. The latter effect correlates with the high-affinity binding of the BH3 peptide to Nr-13 as monitored by the intrinsic tryptophan fluorescence. On the basis of the structural similarity with Bcl-x(L), putative residues involved in this interaction were identified. Nr-13 exhibits a high-affinity interaction with cytochrome c which is prevented by preincubation with the BH3 Bax peptide. These findings are discussed with respect to a model for the regulation of apoptosis in which a direct interaction between the antiapoptotic protein and cytochrome c may prevent the apoptosis. PMID- 12093272 TI - Hydrogen bonding to P700: site-directed mutagenesis of threonine A739 of photosystem I in Chlamydomonas reinhardtii. AB - The primary electron donor P700 of photosystem I is a dimer comprised of chlorophyll a (P(B)) and chlorophyll a' (P(A)). P(A) is involved in a hydrogen bond network with several surrounding amino acid residues and a nearby water molecule. To investigate the influence of hydrogen bond interactions on the properties of P700, the threonine at position A739, which donates a putative hydrogen bond to the 13(1)-keto group of P(A), was replaced with valine, histidine, and tyrosine in Chlamydomonas reinhardtii using site-directed mutagenesis. Growth of the mutants was not impaired. (i) The (P700(+)* - P700) FTIR difference spectra of the mutants lack a negative band at 1634 cm(-1) observed in the wild-type spectrum and instead exhibit a new negative band between 1658 and 1672 cm(-1) depending on the mutation. This band can therefore be assigned to the 13(1)-keto group of P(A) which is upshifted to higher frequencies upon removal of the hydrogen bond. (ii) The main bleaching band in the Q(y)() region of the (P700(+)* - P700) and ((3)P700 - P700) absorption difference spectra is blue shifted for the mutants by approximately 6 nm compared to that of the wild type. A blue shift is also observed for the main bleaching in the Soret region. (iii) The (P700(+)* - P700) CD difference spectrum of the wild type reveals two bands at 694 nm (positive CD) and 680 nm (negative CD) of approximately equal area. For each mutant, these two components are blue-shifted to the same extent. The results strongly suggest that a blue shift of the Q(y) absorption band of P(A) is responsible for a blue shift of the exciton bands. (iv) Redox titrations yielded a decrease in the midpoint potential for the oxidation of P700 by 32 mV for the exchange of Thr against Val. (v) ENDOR spectroscopy shows that the hfc of the methyl protons at position 12 of the spin carrying Chl P(B) is decreased due to the removal of the hydrogen bond to P(A). This indicates a redistribution of spin density in P700(+)* compared to that in the wild type. This gives evidence for an electronic coupling between the two halves of the dimer in the wild type and mutants. PMID- 12093271 TI - Role of His505 in the soluble fumarate reductase from Shewanella frigidimarina. AB - The X-ray structure of the soluble fumarate reductase from Shewanella frigidimarina [Taylor, P., Pealing, S. L., Reid, G. A., Chapman, S. K., and Walkinshaw, M. D. (1999) Nat. Struct. Biol. 6, 1108-1112] clearly shows the presence of an internally bound sodium ion. This sodium ion is coordinated by one solvent water molecule (Wat912) and five backbone carbonyl oxygens from Thr506, Met507, Gly508, Glu534, and Thr536 in what is best described as octahedral geometry (despite the rather long distance from the sodium ion to the backbone oxygen of Met507 (3.1 A)). The water ligand (Wat912) is, in turn, hydrogen bonded to the imidazole ring of His505. This histidine residue is adjacent to His504, a key active-site residue thought to be responsible for the observed pK(a) of the enzyme. Thus, it is possible that His505 may be important in both maintaining the sodium site and in influencing the active site. Here we describe the crystallographic and kinetic characterization of the H505A and H505Y mutant forms of the Shewanella fumarate reductase. The crystal structures of both mutant forms of the enzyme have been solved to 1.8 and 2.0 A resolution, respectively. Both show the presence of the sodium ion in the equivalent position to that found in the wild-type enzyme. The structure of the H505A mutant shows the presence of two water molecules in place of the His505 side-chain which form part of a hydrogen bonding network with Wat48, a ligand to the sodium ion. The structure of the H505Y mutant shows the hydroxyl group of the tyrosine side-chain hydrogen-bonding to a water molecule which is also a ligand to the sodium ion. Apart from these features, there are no significant structural alterations as a result of either substitution. Both the mutant enzymes are catalytically active but show markedly different pH profiles compared to the wild-type enzyme. At high pH (above 8.5), the wild type and mutant enzymes have very similar activities. However, at low pH (6.0), the H505A mutant enzyme is some 20-fold less active than wild-type. The combined crystallographic and kinetic results suggest that His505 is not essential for sodium binding but does affect catalytic activity perhaps by influencing the pK(a) of the adjacent His504. PMID- 12093273 TI - Solution structure of a llama single-domain antibody with hydrophobic residues typical of the VH/VL interface. AB - The three-dimensional structure of a llama single-domain antibody BrucD4-4 was established by use of solution NMR spectroscopy. BrucD4-4 has Val, Gly, Leu, and Trp residues at positions 37, 44, 45, and 47, which are considered to be a hallmark to distinguish llama VH from V(H)H fragments at the germline level. In contrast to the murine and human VHs, BrucD4-4 has sufficient solubility, is monomeric in solution, and displays high-quality NMR spectra characteristic of well-structured proteins. Amide proton/deuterium exchange and the (15)N relaxation data showed that BrucD4-4 has a classic protein structure with a well packed core and comparatively mobile surface loops. The three-dimensional architecture of BrucD4-4 is analogous to that of VHs from murine and human F(v)s and camelid V(H)Hs with two pleated beta-sheets formed by four and five beta strands. A canonical and undistorted beta-barrel exposes a number of hydrophobic residues into the solvent on the surface of the three-dimensional structure. The eight-residue H3 loop folds over the side chain of Val37 similarly to that in llama V(H)Hs; however, this interaction may be transient due to the H3 conformational flexibility. Overall, the surface characteristics of BrucD4-4 with respect to hydrophobicity appear to lie between the human VH domain from Fv Pot and the llama V(H)H fragment HC-V, which may explain its enhanced solubility allowing NMR structural analysis. PMID- 12093274 TI - Analysis of the electron paramagnetic resonance spectrum of a radical intermediate in the coenzyme B(12)-dependent ethanolamine ammonia-lyase catalyzed reaction of S-2-aminopropanol. AB - The structure of the steady-state radical intermediate in the deamination of S-2 aminopropanol catalyzed by ethanolamine ammonia-lyase (EAL) from Salmonella typhimurium has been probed by electron paramagnetic resonance (EPR) spectroscopy using isotopically labeled forms of the substrate and of the adenosylcobalamin cofactor. Electron spin-spin coupling between the radical, centered on the carbon skeleton of the substrate, and the low-spin Co(2+) in cob(II)alamin (B(12r)) produces a dominant splitting of the EPR signals of both the radical and the Co(2+). Analysis of the exchange and dipole-dipole contributions to the spin-spin coupling indicates that the two paramagnetic centers are separated by approximately 11 A. Experiments with (13)C- and with (2)H-labeled forms of S-2 aminopropanol show that the radical is centered on C1 of the carbon skeleton of the substrate in agreement with an earlier report [Babior, B. M., Moss, T. H., Orme-Johnson, W. H., and Beinert, H., (1974) J. Biol. Chem. 249, 4537-4544]. Experiments with perdeutero-S-2-aminopropanol and [2-(15)N]-perdeutero-S-2 aminopropanol reveal a strong hyperfine splitting from the substrate nitrogen, which indicates that the radical is the initial substrate radical created by abstraction of a hydrogen atom from C1 of S-2-aminopropanol. The strong nitrogen hyperfine splitting further indicates that the amino substituent at C2 is approximately eclipsed with respect to the half-occupied p orbital at C1. Experiments with adenosylcobalamin enriched in (15)N in the dimethylbenzimidazole moiety show that the axial base of the cofactor remains attached to the Co(2+) in a functional steady-state reaction intermediate. PMID- 12093275 TI - The structure of L-hydantoinase from Arthobacter aurescens leads to an understanding of dihydropyrimidinase substrate and enantio specificity. AB - L-Hydantoinase from Arthrobacter aurescens (L-Hyd) is a member of the dihydropyrimidinases which in turn belong to the cyclic amidases. Dihydropyrimidinases catalyze the reversible hydrolytic ring opening of dihydropyrimidines as the second step in the catabolism of pyrimidines. In biotechnology, their hydroloytic activity on five-membered cyclic diamides (hydantoins) is used in the enantio-specific production of amino acids from racemic hydantoins. L-Hyd differs from most of the other dihydropyrimidinases by an L-enantio specificity and by lacking activity on possible natural substrates such as dihydropyrimidines. In this paper, we describe the three-dimensional structure of L-Hyd which was solved by molecular replacement using a homology model and subsequently refined to 2.6 A resolution. Each subunit of the tetrameric L-Hyd consists of an elliptically distorted (alpha/beta)(8)-barrel domain, which hosts the active site, and a beta-sheet domain. In the active site, a binuclear zinc center activates a water molecule for nucleophilic attack on the substrates' amide bond. L-Hyd shows a strong homology both in fold and in metal coordination in the active site to another dihydropyrimidinase from Thermus sp. (D-hydantoinase) and to a slightly lesser degree to ureases, dihydroorotase and phosphotriesterase. Using the homology to ureases, a model for the transition state was modeled in the active site of L-Hyd and D-hydantoinase. This model could provide an explanation for the different substrate and enantio selectivities of both dihydropyrimidinases. PMID- 12093276 TI - Target recognition of apocalmodulin by nitric oxide synthase I peptides. AB - An increasing number of proteins are found that are regulated by the Ca(2+)-free state of calmodulin, apocalmodulin. Many of these targets harbor a so-called IQ motif within their primary sequence, but several target proteins of apocalmodulin lack this motif. We investigated whether the Ca(2+)-dependent calmodulin-binding site of nitric oxide synthase I could be transformed into a target site of apocalmodulin. Synthetic peptides representing the wild-type amino acid sequence and several peptides carrying mutations were studied by isothermal titration calorimetry and fluorescence spectroscopy. A single amino acid substitution of a negative charge to a positive charge can convert a classical Ca(2+)-dependent binding site of calmodulin into a target site for apocalmodulin. In addition, the introduction of hydrophobic amino acids increases the apparent binding affinity from the micromolar to the nanomolar range. Binding of wild-type and mutant peptides to Ca(2+)-calmodulin was enthalpically driven, and binding to apocalmodulin was entropically driven. Our data indicate that only a few selected amino acid positions in a calmodulin-binding site determine its Ca(2+) dependency. PMID- 12093277 TI - N(4)C-alkyl-N(4)C cross-linked DNA: bending deformations in duplexes that contain a -CNG- interstrand cross-link. AB - Short DNA duplexes containing a 1,3-N(4)C-alkyl-N(4)C interstrand cross-link that joins the two C residues of a -CNG- sequence were prepared using either a phosphoramidite or convertible nucleoside approach. The alkyl cross-link consists of 2, 4, or 7 methylene groups. The duplexes, which contain a seven-base-pair core and A(3)/T(3) complementary 3'-overhanging ends, were characterized by enzymatic digestion and MALDI-TOF mass spectrometry. Ultraviolet thermal denaturation studies showed that the duplexes denature in a cooperative manner and that the length of the cross-link affects the thermal stability. Thus, the transition temperature of the ethyl cross-linked duplex, 42 degrees C, is 16 degrees C higher than the melting temperature of the corresponding non-cross linked control, whereas the transition temperatures of the butyl and heptyl cross linked duplexes, 73 and 72 degrees C, respectively, are 46-47 degrees C higher. The reduced molecularity of denaturation of the cross-linked duplexes versus melting of the non-cross-linked duplex most likely accounts for these differences. Examination of molecular models suggests that the ethyl cross-link is too short to span the distance between the two C residues at the site of the cross-link in B-form DNA without causing distortion of the helix, whereas less and no distortion would be expected for the butyl and heptyl cross-links, respectively. The circular dichroism spectra, which show greatest deviation in the ethyl cross-linked duplex from B-form DNA, are consistent with this expectation. Anomalous mobilities on native polyacrylamide gels of multimers produced by self-ligation of each of the cross-linked duplexes suggest that the ethyl and butyl cross-linked duplexes undergo bending deformations, whereas multimers derived from the heptyl cross-linked duplex migrated normally. The bending angle was estimated to be 20 degrees, 13 degrees, and 0 degrees for the ethyl, butyl, and heptyl cross-linked duplexes, respectively. Thus, it appears that the degree of bending in these N(4)C-alkyl-N(4)C cross-linked duplexes is controlled by the length of the cross-link. PMID- 12093278 TI - Amplification of the effects of drug resistance mutations by background polymorphisms in HIV-1 protease from African subtypes. AB - The vast majority of HIV-1 infections worldwide are caused by the C and A viral subtypes rather than the B subtype prevalent in the United States and Western Europe. Genomic differences between subtypes give rise to sequence variations in the encoded proteins, including those identified as targets for antiretroviral therapies. In the case of the HIV-1 protease, we reported earlier [Velazquez Campoy et al. (2001) Proc. Natl. Acad. Sci. U.S.A. 98, 6062-6067] that proteases from the C and A subtypes exhibit a higher biochemical fitness in the presence of widely prescribed protease inhibitors. In this paper we present a complete thermodynamic dissection of the differences between proteases from different subtypes and the effects of the V82F/I84V drug-resistant mutation within the framework of the B, C, and A subtypes. These studies involved four inhibitors in clinical use (indinavir, saquinavir, ritonavir, and nelfinavir) and a second generation protease inhibitor (KNI-764). Naturally occurring amino acid polymorphisms found in proteases from the C and A subtypes lower the binding affinities of existing clinical inhibitors by factors ranging between 2 and 7.5 which by themselves are not enough to cause drug resistance. The preexisting lower affinity in the C and A subtypes, however, significantly amplifies the effects of the drug-resistant mutation. Relative to the wild-type B subtype protease, the V82F/I84V drug-resistant mutation within the C and A subtypes lowers the binding affinity of inhibitors by factors ranging between 40 and 3000. When the enzyme kinetic properties (k(cat) and K(m)) are included in the analysis, the biochemical fitness of the C and A subtype drug-resistant mutants can be up to 1000-fold higher than that of the wild-type B subtype protease in the presence of the studied inhibitors. From a thermodynamic standpoint, the combined effects of the drug-resistant mutations and the natural amino acid polymorphisms on the Gibbs energy are additive and involve significant alterations in the enthalpy and entropy changes associated with inhibitor binding. At the biochemical level, the combined effects of naturally existing polymorphisms and drug-resistant mutations might have important consequences on the long-term viability of current HIV-1 protease inhibitors. PMID- 12093279 TI - Affinity-based inhibition of beta-amyloid toxicity. AB - Strategies for interfering with protein aggregation are important for elucidating and controlling the pathologies of amyloid diseases. We have previously identified compounds that block the cellular toxicity of the beta-amyloid peptide, but the relationship between their ability to inhibit toxicity and their affinity for A beta is unknown. To elucidate this relationship, we have developed an assay capable of measuring the affinities of small molecules for beta-amyloid peptide. Our approach employs immobilized beta-amyloid peptide at low density to minimize the problems that arise from variability in the beta-amyloid aggregation state. We found that low-molecular weight (MW of 700-1700) ligands for beta amyloid can be identified readily by using surface plasmon resonance. The best of these bound effectively (K(d) approximately 40 microM) to beta-amyloid. The affinities measured for peptides in the SPR assay correspond to results from A beta cell toxicity assays. The most potent ligands for immobilized beta-amyloid are the most potent inhibitors of the neuronal cell toxicity of beta-amyloid. Compounds with dissociation constants above approximately 100 microM did not show significant activity in the cell toxicity assays. Our data support the hypothesis that ligands exhibiting greater affinity for the beta-amyloid peptide are effective at altering its aggregation and inhibiting cell toxicity. PMID- 12093280 TI - Binding of hemoglobin to red cell membranes with eosin-5-maleimide-labeled band 3: analysis of centrifugation and fluorescence data. AB - We have studied the binding of hemoglobin to the red cell membrane by centrifugation and fluorescence methods. The intact red cell was labeled with eosin-5-maleimide (EM), which specifically reacts with lysine 430 of band 3. Even though this residue is not part of the cytoplasmic domain of band 3 (cdb3) associated with hemoglobin binding, fluorescence quenching was observed when hemoglobin bound to inside-out vesicles (IOVs). The use of fluorescence quenching to measure band 3 binding was quantitatively compared with the binding determined by centrifugation, which measures binding to band 3 and non-band 3 sites. For the centrifugation it was necessary to include the non-band 3 association constants determined from chymotrypsin-treated IOVs. The binding of hemoglobin to band 3 was interpreted in terms of the binding of two hemoglobin tetramers to each band 3 dimer. An anticooperative interaction associated with the conformational change produced when hemoglobin binds results in a 2.8-fold decrease in the intrinsic constant of (1.54 +/- 0.25) x 10(7) M(-1) for the binding of the second hemoglobin molecule. From the changes in lifetime produced by binding the first and second hemoglobin molecules, it was possible to show that the conformational change associated with binding the second hemoglobin molecule results in a decrease of the heme-eosin distance from 47.90 to 44.78 A. Reaction of cyanate with the alpha-amino group of hemoglobin (HbOCN) is shown to produce a very dramatic decrease in the binding of hemoglobin to both the band 3 and non-band 3 sites. The intrinsic constant for binding the first hemoglobin molecule to band 3 decreases by a factor of 29 to (5.34 +/- 0.15) x 10(5) M(-1). The anticooperative interaction is greater with the intrinsic constant decreasing by a factor of 3.8 for the binding of the second hemoglobin tetramer to band 3. In addition, the nature of the conformational change produced by binding hemoglobin is very different with the second HbOCN increasing the heme-eosin distance to 55.99 A. The utilization of eosin-5-maleimide-reacted red cell membrane to study hemoglobin binding makes it possible to directly study the binding to band 3. At the same time a sensitive probe of the conformational changes, which occur when hemoglobin binds to band 3, is provided. PMID- 12093281 TI - Deamidation in human gamma S-crystallin from cataractous lenses is influenced by surface exposure. AB - A major component of human nuclear cataracts is water-insoluble, high molecular weight protein. A significant component of this protein is disulfide bonded gamma S-crystallin that can be reduced to monomers by dithiothreitol. Analysis of this reduced gamma S-crystallin showed that deamidation of glutamine and asparagine residues is a principal modification. Deamidation is one of the modifications of lens crystallins associated with aging and cataractogenesis. One proposed hypothesis of cataractogenesis is that it develops in response to altered surface charges that cause conformational changes, which, in turn, permit formation of disulfide bonds and crystallin insolubility. This report, showing deamidation among the disulfide bonded gamma S-crystallins from cataractous lenses, supports this hypothesis. PMID- 12093282 TI - Site-directed mutagenesis and chemical modification of the six native cysteine residues of the rat mitochondrial carnitine carrier: implications for the role of cysteine-136. AB - By use of site-directed mutagenesis in combination with chemical modification of mutated proteins, the role of the six Cys residues in the transport function of the rat mitochondrial carnitine carrier (CAC) was studied. Several CAC mutants, in which one or more Cys residues had been replaced with Ser, were overexpressed in Escherichia coli, purified, and reconstituted in liposomes. The efficiency of incorporation into liposomes of the reconstituted proteins was lower for all constructs lacking Cys-23. Single, double, and quadruple replacement mutants showed V(max) comparable to that of the wild type. On the basis of the values of internal and external transport affinities (K(m)) for carnitine and of their comparison with those measured in mitochondria, the recombinant CAC is oriented unidirectionally in the liposomes, right side out compared to mitochondria. Substitution of Cys-136 with Ser caused a nearly complete loss of sensitivity of the CAC to N-ethylmaleimide, (2-aminoethyl)methanethiosulfonate hydrobromide (MTSES), and other hydrophilic SH reagents but not to the very hydrophobic N phenylmaleimide. The wild-type CAC and the mutants containing Cys-136 showed substrate protection against NEM and MTSES inhibition and against NEM labeling. The data show that none of the native cysteines is essential for the transport mechanism and that Cys-136 is the major target of SH reagents and raise the hypothesis that Cys-136 is accessible from the external medium and is located at, or near, the substrate binding site. A model of the CAC is proposed in which the matrix hydrophilic loop containing Cys-136 protrudes into the membrane between the transmembrane domains of the protein. PMID- 12093283 TI - Brain-specific p25 protein binds to tubulin and microtubules and induces aberrant microtubule assemblies at substoichiometric concentrations. AB - Previously, we have demonstrated the presence of a protein factor [tubulin polymerization perturbing protein (TPPP)] in brain and neuroblastoma cell but not in muscle extract that uniquely influences the microtubule assembly. Here we describe a procedure for isolation of this protein from the cytosolic fraction of bovine brain and present evidence that this protein is a target of both tubulin and microtubules in vitro. The crucial step of the purification is the cationic exchange chromatography; the bound TPPP is eluted at high salt concentrations, indicating the basic character of the protein. By IDA-nanoLC-MS analysis of the peptides extracted from the gel-digested purified TPPP, we show the presence of a single protein in the purified fraction that corresponds to p25, a brain-specific protein the function of which has not been identified. Circular dichroism data have revealed that, on one hand, the alpha-helix content of p25 is very low (4%) with respect to the predicted values (30-43%), and its binding to tubulin induces remarkable alteration in the secondary structure of the protein(s). As shown by turbidimetry, pelleting experiments, and electron microscopy, p25 binds to paclitaxel-stabilized microtubules and bundles them. p25 induces formation of unusual (mainly double-walled) microtubules from tubulin in the absence of paclitaxel. The amount of aberrant tubules formed depends on the p25 concentration, and the process occurs at substoichiometric concentrations. Our in vitro data suggest that p25 could act as a unique MAP in vivo. PMID- 12093284 TI - The biosynthetic incorporation of short-chain linear saturated fatty acids by Acholeplasma laidlawii B may suppress cell growth by perturbing membrane lipid polar headgroup distribution. AB - Acholeplasma laidlawii B cells made fatty acid auxotrophic by growth in the presence of the biotin-binding agent avidin grow increasingly poorly at 37 degrees C when supplemented with single exogenous linear saturated fatty acids of decreasing hydrocarbon chain length. Interestingly, this progressive decrease in growth yields with decreasing hydrocarbon chain length is not observed when cells are cultured in the presence of other classes of exogenous fatty acids. Moreover, normal growth is observed is other types of fatty acids with equivalent or shorter hydrocarbon chain lengths, indicating that poor growth in the presence of short-chain linear saturated fatty acids cannot be due to a decrease in membrane lipid bilayer thickness per se. To understand the molecular basis of such growth inhibition, we determined the growth yields, membrane lipid fatty acid and polar headgroups compositions, and phase state and fluidity of the membrane lipids in cells progressively biosynthetically enriched in tridecanoic acid (13:0) or dodecanoic acid (12:0). The growth of fatty acid auxotrophic A. laidlawii B cells grown in the presence of binary combinations of an exogenous fatty acid which supports normal growth on its own and 13:0 or 12:0 revealed that growth inhibition is not observed until 13:0 and 12:0 biosynthetic incorporation levels reach about 90 and 60 mol %, respectively, after which growth is markedly inhibited. Differential scanning calorimetric analyses of membranes from cells maximally enriched in 13:0 indicate that the lipid gel/liquid-crystalline phase transition temperature is unexpectedly high but that at the growth temperature of 37 degrees C, the membrane lipid bilayer is almost exclusively in the liquid crystalline state but is certainly not excessively fluid. However, high levels of 13:0 incorporation produce a greatly elevated level of the high melting, reversed nonlamellar phase-preferring lipid component monoglucosyl diacylglycerol, and greatly reduced levels of all other membrane lipid components. This marked elevation of monoglucosyl diacylglycerol levels can be rationalized as a regulatory response which maintains the lamellar/nonlamellar phase-forming propensity of the total membrane lipid mixture relatively constant in the face of the biosynthetic incorporation of increasing quantities of short-chain saturated fatty acids, which favor the lamellar phase. However, this lipid biosynthetic response produces a marked decline in the levels of anionic phospholipid and phosphoglycolipid which are probably required to maintain the minimal negative surface charge density of the lipid bilayer, which we suggest is responsible for the observed growth inhibition. This work shows that the lipid biosynthetic regulatory mechanisms present in this organism may sometimes operate at cross purposes such that it is not possible to simultaneously optimize all of the biologically relevant physical properties of the membrane lipid bilayer. PMID- 12093286 TI - Bistranded oxidized purine damage clusters: induced in DNA by long-wavelength ultraviolet (290-400 nm) radiation? AB - Bistranded clustered DNA damages involving oxidized bases, abasic sites, and strand breaks are produced by ionizing radiation and radiomimetic drugs, but it was not known whether they can be formed by other agents, e.g., nonionizing radiation. UV radiation produces clusters of cyclobutyl pyrimidine dimers, photoproducts that occur individually in high yield. Since long-wavelength UV (290-400 nm) radiation induces oxidized bases, abasic sites, and strand breaks at low yields, we tested whether it also produces clusters containing these lesions. We exposed supercoiled pUC18 DNA to UV radiation with wavelengths of >290 nm (UVB plus UVA radiation), and assessed the induction of bistranded clustered oxidized purine and abasic clusters, as recognized by Escherichia coli Fpg protein and E. coli Nfo protein (endonuclease IV), respectively, as well as double-strand breaks. These three classes of bistranded clusters were detected, albeit at very low yields (37 Fpg-OxyPurine clusters Gbp(-1) kJ(-1) m(2), 8.1 double-strand breaks Gbp(-1) kJ(-1) m(2), and 3.4 Nfo-abasic clusters Gbp(-1) kJ(-1) m(2)). Thus, these bistranded OxyPurine clusters, abasic clusters, and double-strand breaks are not uniquely induced by ionizing radiation and radiomimetic drugs, but their level of production by UVB and UVA radiation is negligible compared to the levels of frequent photoproducts such as pyrimidine dimers. PMID- 12093285 TI - DNA sequence recognition by the antitumor drug ditercalinium. AB - The antitumor drug ditercalinium is a rare example of a noncovalent DNA-binding ligand that forms bisintercalation complexes via the major groove of the double helix. Previous structural studies have revealed that the two connected pyridocarbazolium chromophores intercalate into DNA with the positively charged bis(ethylpiperidinium) linking chain oriented to the wide groove side of the helix. Although the interaction of ditercalinium with short oligonucleotides containing 4-6 contiguous GC base pairs has been examined in detail by biophysical and theoretical approaches, the sequence preference for ditercalinium binding to long DNA fragments that offer a wide variety of binding sites has been investigated only superficially. Here we have investigated both sequence preferences and possible molecular determinants of selectivity in the binding of ditercalinium to DNA, primarily using methods based upon DNase I footprinting. A range of multisite DNA substrates, including several natural restriction fragments and different PCR-generated fragments containing unconventional bases (2,6-diaminopurine, inosine, uridine, 5-fluoro- and 5-methylcytosine, 7 deazaguanine, 7-deazaadenine, and N(7)-cyanoboranoguanine), have been employed to show that ditercalinium selectively recognizes certain GC-rich sequences in DNA and to identify some of the factors which affect its DNA-binding sequence selectivity. Specifically, the footprinting data have revealed that the 2-amino group on the purines or the 5-methyl group on the pyrimidines is not essential for the formation of ditercalinium-DNA complexes whereas the major groove oriented N(7) of guanine does appear as a key element in the molecular recognition process. The loss of N(7) at guanines but not adenines is sufficient to practically abolish sequence-selective binding of ditercalinium to DNA. Thus, as expected for a major groove binding drug, the N(7) of guanine is normally required for effective complex formation with GC base pairs, but interestingly the substitution of the N(7) with a relatively bulky cyanoborane group does not markedly affect the sequence recognition process. Therefore, the hydrogen bond accepting capability at N(7) of guanines is not sufficient to explain the GC selective drug-DNA association, and the implications of these findings are considered. PMID- 12093287 TI - Repair of oligodeoxyribonucleotides by O(6)-alkylguanine-DNA alkyltransferase. AB - Activity of the DNA repair protein O(6)-alkylguanine-DNA alkyltransferase (AGT) is an important source of tumor cell resistance to alkylating agents. AGT inhibitors may prove useful in enhancing chemotherapy. AGT is inactivated by reacting stoichiometrically with O(6)-benzylguanine (b(6)G), which is currently in clinical trials for this purpose. Short oligodeoxyribonucleotides containing a central b(6)G are more potent inactivators of AGT than b(6)G. We examined whether human AGT could react with oligodeoxyribonucleotides containing multiple b(6)G residues. The single-stranded 7-mer 5'-d[T(b(6)G)(5)G]-3' was an excellent AGT substrate with all five b(6)G adducts repaired although one adduct was repaired much more slowly. The highly b(6)G-resistant Y158H and P140K AGT mutants were also inactivated by 5'-d[T(b(6)G)(5)G]-3'. Studies with 7-mers containing a single b(6)G adduct showed that 5'-d[TGGGG(b(6)G)G]-3' was more poorly repaired by wild-type AGT than 5'-d[T(b(6)G)GGGGG]-3' and 5'-d[TGG(b(6)G)GGG]-3' and was even less repairable by mutants Y158H and P140K. This positional effect was unaffected by interchanging the terminal 5'- or 3'-nucleotides and was also observed with single-stranded 16-mer oligodeoxyribonucleotides containing O(6) methylguanine, where a minimum of four nucleotides 3' to the lesion was required for the most efficient repair. Annealing with the reverse complementary strands to produce double-stranded substrates increased the ability of AGT to repair adducts at all positions except at positions 2 and 15. Our results suggest that AGT recognizes the polarity of single-stranded DNA, with the best substrates having an adduct adjacent to the 5'-terminal residue. These findings will aid in designing novel AGT inhibitors that incorporate O(6)-alkylguanine adducts in oligodeoxyribonucleotide contexts. PMID- 12093288 TI - Isolation, size estimates, and spectral heterogeneity of an oligomeric series of light-harvesting 1 complexes from Rhodobacter sphaeroides. AB - A series of light-harvesting 1 (LH1) complexes was isolated by lithium dodecyl sulfate-polyacrylamide gel electrophoresis at 4 degrees C from Rhodobacter sphaeroides M21, which lacks the peripheral light-harvesting 2 (LH2) complex. This ladder of LH1 bands was also demonstrated in the wild type, partially superimposed upon a smaller number of LH2 complexes. An assessment of electrophoretic mobility vs acrylamide concentration, in which the reaction center LM particle and annular LH1 and LH2 complexes were used as standards of known structure, indicated that the LH1 gel bands 2 to 10 represent regular oligomers of an alpha beta heterodimeric unit, that vary in size from (alpha beta)(2-3) to (alpha beta)(10-11). The isolated LH1 complexes exhibited oligomeric state dependent optical properties, characterized by red shifts in near-IR absorption and emission maxima at 77 K of approximately 6 nm as aggregate sizes increased from approximately 3 to 7-8 alpha beta-heterodimers, accompanied by shifts in highly polarized fluorescence from the blue to the red side of the absorption band. This has been explained by the oligomerization of heterodimers to form a curvilinear array of excitonically coupled chromophores, with the anisotropic long-wavelength component, designated originally as B896, corresponding to low energy excitonic transitions arising from interactions within inhomogeneous BChl clusters [Westerhuis et al. (1999) J. Phys. Chem. B 103, 7733-7742]. Differences in electrophoretic profiles of LH1 bands between strains M21 and M2192, an LH1-only strain that also lacks PufX, further suggested that the more rapidly migrating bands represent arced fragments of the curvilinear array of LH1 complexes thought to exist as a large closed circular structure only in the latter strain. The electrophoretic banding pattern also indicated that the LH1 complex may be located at the peripheries of dimeric intramembrane particle arrays seen in freeze-fracture replicas of tubular M21 membranes; the possible role for the PufX protein in the assembly of these structures is discussed. PMID- 12093289 TI - (13)C MAS NMR and photo-CIDNP reveal a pronounced asymmetry in the electronic ground state of the special pair of Rhodobacter sphaeroides reaction centers. AB - Reaction centers of wild-type Rhodobacter sphaeroides were selectively (13)C isotope labeled in bacteriochlorophyll and bacteriopheophytin. (13)C solid-state CP/MAS NMR and photo-CIDNP were used to provide insight into the electronic structure of the primary electron donor and acceptor on the atomic scale. The first 2-dimensional photochemically induced dynamic nuclear polarization (photo CIDNP) (13)C-(13)C solid-state MAS NMR spectra reveal that negative charging of the two BChl rings of the primary donor is involved in ground-state tuning of the oxidation potential of these cofactors in the protein via local electrostatic interactions. In particular, the (13)C shifts show moderate differences in the electronic structure between the two BChl molecules of the special pair in the electronic ground state, which can be attributed to hydrogen bonding of one of the BChl molecules. The major fraction of the electron spin density is strongly delocalized over the two BChl molecules of the special pair and the photochemically active BPhe. A small fraction of the pi-spin density is distributed over a fourth component, which is assigned to the accessory BChl. Comparison of the photo-CIDNP data with "dark" NMR spectra obtained in ultra high field indicates a rigid special pair environment upon photoreaction and suggests that structural changes of the aromatic macrocycles of the two BChl molecules of the special pair do not significantly contribute to the reorganization energy associated with the charge-transfer process. PMID- 12093290 TI - Tuning the reduction potential of engineered cytochrome c-553. AB - Cytochrome c-553 from Desulfovibrio vulgaris exhibits a highly exposed heme and an unusually low reduction potential with respect to other c-type cytochromes. Solvent heme exposure has been indicated as one of the most important factors in modulating the midpoint potential of the redox center. To test this hypothesis, a unique surface-exposed cysteine has been substituted for either M23 or G51 to produce the corresponding mutants and allow the formation of homodimers through a specific disulfide bridge. The reduction potentials, determined via spectroelectrochemistry, show an increase from +20 +/- 5 mV for the wt to +88 +/- 5 and +105 +/- 5 mV for the M23C-M23C homodimer and G51C-G51C homodimer, respectively. Chemical denaturation of the homodimers leads to parameters related to the hydrophobicity (m) and the number of buried side chains (n(B)), which suggest a decrease of exposure of the heme as a result of dimerization. These results are consistent with the heme-accessible surface area (ASA) calculated from a computer model of the homodimers. The ASA values show a decrease from 73 A(2) for the wt to 66 and 50 A(2) per heme for the M23C-M23C homodimer and G51C G51C homodimer, respectively. The trend of the m- and n(B)-values, the degree of solvent accessibility, and the midpoint potential observed upon formation of the homodimers indicate a correlation between the reduction potential values and the exclusion of water from the heme surface. PMID- 12093291 TI - The common gamma chain (gamma c) is a required signaling component of the IL-21 receptor and supports IL-21-induced cell proliferation via JAK3. AB - The common cytokine receptor gamma chain (gamma c), an essential component of the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15, is critical for the development and function of lymphocytes. Recently, a novel lymphokine (IL-21) and its receptor (IL-21R alpha) were described which profoundly affect the growth and activation state of B, T, and NK cells in concert with other lymphokines or stimuli [Parrish-Novak, J., et al. (2000) Nature 408, 57-63]. In this report, we show that gamma c is also a required signaling component of the IL-21 receptor (IL-21R) using the gamma c-deficient X-linked severe combined immunodeficiency (XSCID) lymphoblastoid cell line JT, and JT cells reconstituted with gamma c (JT/gamma c). Moreover, we demonstrate a functional requirement for both gamma c and the gamma c-associated Janus family tyrosine kinase 3 (JAK3) in IL-21-induced proliferation of pro-B-lymphoid cells engineered to express human IL-21R alpha (BaF3/IL-21R alpha). Retroviral-mediated transduction of wild-type gamma c into XSCID JT cells restored function to the IL-21R, as shown by IL-21-induced tyrosine phosphorylation of JAK1 and JAK3, and downstream activation of STAT5, in JT/gamma c cells as well as BaF3/IL-21R alpha and primary splenic B cells. In contrast, IL-21 failed to activate the JAK-STAT pathway in nonreconstituted JT cells. Monoclonal antibodies specific for the gamma c chain effectively inhibited IL-21-induced growth of BaF3/IL-21R alpha cells, supporting a functional role for this molecule in the IL-21R complex. In addition, the specific JAK3 tyrosine kinase inhibitor WHI-P131 significantly reduced IL-21-induced proliferation of BaF3/IL-21R alpha cells. Taken together, these results definitively demonstrate that IL-21-mediated signaling requires the gamma c chain, and indicate that JAK3 is an essential transducer of gamma c-dependent survival and/or mitogenic signals induced by this cytokine. PMID- 12093292 TI - Role of the N-terminus of epidermal growth factor in ErbB-2/ErbB-3 binding studied by phage display. AB - Epidermal growth factor (EGF) binds with high affinity to the EGF receptor, also known as ErbB-1, but upon replacement of the N-terminal linear region by neuregulin (NRG) 1 or transforming growth factor (TGF) alpha sequences it gains in addition high affinity for ErbB-2/ErbB-3 heterodimers. However, these chimeras weakly bind to ErbB-3 alone. To further dissect the ligand binding selectivity of the ErbB network, we have applied the phage display technique to examine the role of the linear N-terminal region in EGF for interaction with ErbB-2/ErbB-3 heterodimers. A library of EGF variants was constructed in which residues 2, 3, and 4 were randomly mutated, followed by selection for binding to intact MDA-MB 453 cells that overexpress ErbB-2 and ErbB-3 but lack ErbB-1. Analysis of the selected phage EGF variants revealed clones with high binding affinity to ErbB 2/ErbB-3 while maintaining high affinity to ErbB-1. In these variants, Trp (or alternatively His) was almost exclusively present at position 2, while specific combinations of hydrophobic, basic, and small residues were found at positions 3 and 4. The mitogenic activity of the phage EGF variants corresponded with their relative binding affinity. Two of the selected EGF variants, EGF/WVS and EGF/WRS, were further characterized as recombinant proteins. In contrast to previously characterized chimeras of EGF with NRG-1 or TGF-alpha, these variants did not only show high binding affinity for ErbB-2/ErbB-3 heterodimers but also for ErbB 3 alone. These data show that the linear N-terminal region of EGF-like growth factors is directly involved in binding to ErbB-3. PMID- 12093293 TI - Specificity of memapsin 1 and its implications on the design of memapsin 2 (beta secretase) inhibitor selectivity. AB - Memapsin 1 is closely homologous to memapsin 2 (BACE), or beta-secretase, whose action on beta-amyloid precursor protein (APP) leads to the production of beta amyloid (A beta) peptide and the progression of Alzheimer's disease. Memapsin 2 is a current target for the development of inhibitor drugs to treat Alzheimer's disease. Although memapsin 1 hydrolyzes the beta-secretase site of APP, it is not significantly present in the brain, and no direct evidence links it to Alzheimer's disease. We report here the residue specificity of eight memapsin 1 subsites. In substrate positions P(4), P(3), P(2), P(1), P(1)', P(2)', P(3)', and P(4)', the most preferred residues are Glu, Leu, Asn, Phe, Met, Ile, Phe, and Trp, respectively, while the second preferred residues are Gln, Ile, Asp, Leu, Leu, Val, Trp, and Phe, respectively. Other less preferred residues can also be accommodated in these subsites of memapsin 1. Despite the broad specificity, these residue preferences are strikingly similar to those of human memapsin 2 [Turner et al. (2001) Biochemistry 40, 10001-10006] and thus pose a serious problem to the design of differentially selective inhibitors capable of inhibiting memapsin 2. This difficulty was confirmed by the finding that several potent memapsin 2 inhibitors effectively inhibited memapsin 1 as well. Several possible approaches to overcome this problem are discussed. PMID- 12093294 TI - Comparison of rates and kinetic isotope effects using PEG-modified variants and glycoforms of glucose oxidase: the relationship of modification of the protein envelope to C-H activation and tunneling. AB - An earlier investigation of the temperature dependencies of rates and kinetic isotope effects (KIEs) in glucose oxidase (GO) used variants that differed in the extent of glycosylation at the surface of the protein. Kohen et al. [Kohen, A., Jonsson, T., and Klinman, J. P. (1997) Biochemistry 36, 2603-2611] presented evidence that the KIE on the Arrhenius prefactor varied as a function of protein modification, concluding that the degree of hydrogen tunneling at the active site was dependent on changes in mass at the surface. We now examine GO proteins containing polyethylene glycol (PEG) at their surface and a more extensively glycosylated form of GO, to distinguish simple mass effects from other sources of altered catalytic behavior. One PEG variant was created by modifying deglycosylated GO with short PEG chains (average of 350 Da each), while another contained a smaller number of long PEG chains (average of 5000 Da each). The light (146 kDa) and heavy (211 kDa) PEG variants and the hyperglycosylated variant display isotope effects on the Arrhenius prefactor that are similar (A(D)/A(T) = 0.55-0.62), while the unperturbed wild-type GO (WT-GO) is found to have an A(D)/A(T) that is reassessed as being close to unity. It appears that any modification of the protein surface away from that of the wild type gives rise to altered behavior for hydrogen transfer in the active site. We have also compared the effect of enthalpies of activation on both k(cat)/K(M) and k(cat) for the variants, introducing a new method to extract the k(cat)/K(M) rate constant and enthalpy of activation for the tritiated substrate from competitive KIE experiments. We find similar trends in Delta H(++) for both competitive and noncompetitive parameters and a smaller trend in k(cat) than reported earlier. Correlations are observed between A(D)/A(T) and both the enthalpies of activation and the thermal melt temperatures (T(M)) of the GO isoforms. In addition to the present study, there are now a number of examples where a perturbation of enzyme structure away from that of the wild type causes the observed KIE to become more temperature-dependent. The implications of these findings are discussed in the context of hydrogen tunneling and the relationship of protein structure and dynamics to this process. PMID- 12093295 TI - Mutagenesis of Trp(54) and Trp(203) residues on Fibrobacter succinogenes 1,3-1,4 beta-D-glucanase significantly affects catalytic activities of the enzyme. AB - The possible structural and catalytic functions of the nine tryptophan amino acid residues, including Trp(54), Trp(105), Trp(112), Trp(141), Trp(148), Trp(165), Trp(186), Trp(198), and Trp(203) in Fibrobacter succinogenes 1,3-1,4-beta-D glucanase (Fs beta-glucanase), were characterized using site-directed mutagenesis, initial rate kinetics, fluorescence spectrometry, and structural modeling analysis. Kinetic studies showed that a 5-7-fold increase in K(m) value for lichenan was observed for W141F, W141H, and W203R mutant Fs beta-glucanases, and approximately 72-, 56-, 30-, 29.5-, 4.9-, and 4.3-fold decreases in k(cat) relative to that for the wild-type enzyme were observed for the W54F, W54Y, W141H, W203R, W141F, and W148F mutants, respectively. In contrast, W186F and W203F, unlike the other 12 mutants, exhibited a 1.4- and 4.2-fold increase in k(cat), respectively. W165F and W203R were the only two mutants that exhibited a 4-7-fold higher activity relative to the wild-type enzyme after they were incubated at pH 3.0 for 1 h. Fluorescence spectrometry indicated that all of the mutations on the nine tryptophan amino acid residues retained a folding similar to that of the wild-type enzyme. Structural modeling and kinetic studies suggest that Trp(54), Trp(141), Trp(148), and Trp(203) play important roles in maintaining structural integrity in the substrate-binding cleft and the catalytic efficiency of the enzyme. PMID- 12093296 TI - Identification of a novel pyridoxal 5'-phosphate binding site in adenosylcobalamin-dependent lysine 5,6-aminomutase from Porphyromonas gingivalis. AB - Lysine 5,6-aminomutase (5,6-LAM) catalyzes the interconversion of D-lysine with 2,5-diaminohexanoate and of L-beta-lysine with 3,5-diaminohexanoate. The coenzymes for 5,6-LAM are adenosylcobalamin (AdoCbl) and pyridoxal 5'-phosphate (PLP). In the proposed chemical mechanism, AdoCbl initiates the formation of substrate radicals, and PLP facilitates the radical rearrangement by forming an external aldimine linkage with the epsilon-amino group of a substrate, either D lysine or L-beta-lysine. In the resting enzyme, an internal aldimine between PLP and an essential lysine in the active site facilitates productive PLP binding and catalysis. We present here biochemical, biophysical, and site-directed mutagenesis experiments, which document the existence of an essential lysine residue in the active site of 5,6-LAM from Porphyromonas gingivalis. Reduction of 5,6-LAM with NaBH(4) rapidly inactivates the enzyme and shifts the electronic absorption band from 420 to 325 nm. This is characteristic of the reduction of an aldimine linkage between the carbonyl group of PLP and the epsilon-amino group of a lysine residue. The reduced peptide was identified by Q-TOF/MS and further confirmed by Q-TOF/MS/MS sequencing. We show that lysine 144 in the small subunit of 5,6-LAM is the essential lysine residue. Lysine 144(beta) is separated by only 11 amino acids from histidine 133(beta), which forms a part of the "base-off" AdoCbl binding motif. The sequence of the novel PLP-binding motif is conserved in 5,6-LAM from Clostridium sticklandii and P. gingivalis, and it is distinct from all known PLP-binding motifs. Mutation of lysine 144(beta) to glutamine led to K144Q(beta)-5,6-LAM, which displayed no enzymatic activity and no absorption band corresponding to an internal PLP-aldamine. In summary, we introduce a novel PLP binding motif, the first to be discovered in an AdoCbl-dependent enzyme. PMID- 12093297 TI - Chemical transformation is not rate-limiting in the reaction catalyzed by Escherichia coli 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase. AB - 6-Hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK) catalyzes the transfer of pyrophosphate from ATP to 6-hydroxymethyl-7,8-dihydropterin (HMDP). Because HPPK is essential for microorganisms but is absent from human and animals, the enzyme is an excellent target for developing antimicrobial agent. Thermodynamic analysis shows that Mg(2+) is important not only for the binding of nucleotides but also for the binding of HMDP. Transient kinetic analysis shows that a step or steps after the chemical transformation are rate-limiting in the reaction catalyzed by HPPK. The pre-steady-state kinetics is composed of a burst phase and a steady-state phase. The rate constant for the burst phase is approximately 50 times larger than that for the steady-state phase. The latter is very similar to the k(cat) value measured by steady-state kinetics. A set of rate constants for the individual steps of the HPPK-catalyzed reaction has been determined by a combination of stopped-flow and quench-flow analyses. These results form a thermodynamic and kinetic framework for dissecting the roles of active site residues in the substrate binding and catalysis by HPPK. PMID- 12093300 TI - Standardisation of costs: the Dutch Manual for Costing in economic evaluations. AB - The lack of a uniform costing methodology is often considered a weakness of economic evaluations that hinders the interpretation and comparison of studies. Standardisation is therefore an important topic within the methodology of economic evaluations and in national guidelines that formulate the formal requirements for studies to be considered when deciding on the reimbursement of new medical therapies. Recently, the Dutch Manual for Costing: Methods and Standard Costs for Economic Evaluations in Health Care (further referred to as "the manual") has been published, in addition to the Dutch guidelines for pharmacoeconomic research. The objectives of this article are to describe the main content of the manual and to discuss some key issues of the manual in relation to the standardisation of costs. The manual introduces a six-step procedure for costing. These steps concern: the scope of the study;the choice of cost categories;the identification of units;the measurement of resource use;the monetary valuation of units; andthe calculation of unit costs. Each step consists of a number of choices and these together define the approach taken. In addition to a description of the costing process, five key issues regarding the standardisation of costs are distinguished. These are the use of basic principles, methods for measurement and valuation, standard costs (average prices of healthcare services), standard values (values that can be used within unit cost calculations), and the reporting of outcomes. The use of the basic principles, standard values and minimal requirements for reporting outcomes, as defined in the manual, are obligatory in studies that support submissions to acquire reimbursement for new pharmaceuticals. Whether to use standard costs, and the choice of a particular method to measure or value costs, is left mainly to the investigator, depending on the specific study setting. In conclusion, several instruments are available to increase standardisation in costing methodology among studies. These instruments have to be used in such a way that a balance is found between standardisation and the specific setting in which a study is performed. The way in which the Dutch manual tries to reach this balance can serve as an illustration for other countries. PMID- 12093299 TI - Homocyst(e)ine and coronary heart disease: pharmacoeconomic support for interventions to lower hyperhomocyst(e)inaemia. AB - Homocyst(e)ine, a sulphur-containing amino acid, is an intermediate formed during the metabolism of the essential amino acid methionine. Biological and epidemiological evidence suggest that elevated plasma levels of homocyst(e)ine are a risk factor for atherosclerosis and coronary heart disease (CHD). In the general US population, hyperhomocyst(e)inaemia is common and most often due to mild nutritional deficiencies in the B vitamins (folic acid, vitamin B(12) and vitamin B(6)). While high homocyst(e)ine levels can be effectively lowered using folic acid and other B vitamins, it is unknown whether such vitamin therapy will lead to clinical benefits. Given that strategies for homocyst(e)ine-lowering are safe and inexpensive, however, even small reductions in CHD risk will be highly cost effective. Thus, it may be prudent for patients to ensure an adequate daily intake of dietary folic acid and other B vitamins and for physicians to screen high-risk adults such as those with established CHD as we await definitive results from ongoing clinical trials. PMID- 12093301 TI - Health-related quality of life and cost impact of irritable bowel syndrome in a UK primary care setting. AB - OBJECTIVES: To identify the impact of irritable bowel syndrome (IBS) on health related quality of life (HR-QOL), time off work and the utilisation and cost of health services. DESIGN: A case-control study was undertaken matching patients with IBS and controls. Quality-of-life information was collected using the Medical Outcomes Study 36-item Short Form (SF-36) health survey, EuroQOL instrument (EQ-5D) and IBS Quality-of-Life (IBS-QOL) instruments. Data on time off work was also collected. National Health Service (NHS) resource use in primary and secondary care was estimated by review of general practitioner (GP) and hospital records over a 12-month period. SETTING: Recruitment was from six GPs' surgeries in the Trent Region of the United Kingdom. PARTICIPANTS: 161 patients with IBS, as defined by the Rome Criteria I were recruited. These were compared with 213 controls matched for age, sex and social characteristics. MAIN OUTCOME MEASURES: SF-36 and EQ-5D scores; mean number of days off work; mean NHS costs per person during the 12-month study period. RESULTS: Patients with IBS had considerably lower HR-QOL than controls. They scored worse in all dimensions of the SF-36 and the EQ-5D and they had more time off work. On average patients with IBS cost the NHS 123 UK pounds (95% confidence interval: 35 UK pounds to 221 UK pounds, 1999 values) more per year than individuals in the control group (p = 0.04). CONCLUSIONS: IBS affects patients through reduced quality of life, more time off work and greater healthcare utilisation than a control group of patients without IBS. The difference in quality of life was pronounced and unusual in that it was influential in every dimension of both the SF-36 and the EQ-5D. PMID- 12093302 TI - Limitations of the methods used for calculating quality-adjusted life-year values. AB - OBJECTIVE: To test the validity of the techniques used to calculate quality adjusted life-year (QALY) values based on utility functions, using a real population dataset. DESIGN: Using the standard gamble technique, we gathered the preferences of a population sample of 189 individuals on a combination of probabilities concerning four simple health states (no physical disability, limp, walk with crutches and need a wheelchair), and three life-year spans (5, 10 and 15 years). Each of the four assumptions of the multiplicative model was tested based on the results of the experiment. RESULTS: The utility of the health state "limp" was high at 0.89 and that of "walk with crutches" only slightly lower at 0.85. However, of the 189 individuals, only 57 are not in contradiction with the assumption of mutual utility independence since they strictly preferred (15 years, "need a wheelchair") over (10 years, "need a wheelchair") and (15 years, "need a wheelchair") over (5 years, "need a wheelchair"). For these 57 individuals, the results of this study do not fit the assumptions underpinning the multiplicative model. CONCLUSION: This work suggests that the techniques used as a basis from which to calculate QALY values are flawed. In particular, the underlying assumptions of the multiattribute utility model do not correspond to behaviour patterns observed in a real population. It therefore appears that use of the QALY technique should be questioned in healthcare decision-making settings. PMID- 12093303 TI - A model to estimate the cost benefit of an occupational vaccination programme for influenza with Influvac in the UK. AB - OBJECTIVES: To model the economic impact of introducing an occupational vaccination programme for influenza with an inactivated influenza subunit vaccine (Influvac) in the UK. DESIGN AND SETTING: Using published sources, a decision tree was constructed which modelled the costs and benefits of introducing an influenza vaccine in a business in the UK from the perspective of an employer. STUDY PARTICIPANTS AND INTERVENTIONS: The model considered the implementation of an occupational vaccination programme with Influvac in a business employing 1000 normal healthy adults earning the national average wage in the UK. The model assumed that 95% of employees would be absent from work after contracting influenza for a mean of 5 days and that the level of productivity would be reduced by 60% for one day by 85% of sick employees returning to work. MAIN OUTCOME MEASURES AND RESULTS: The expected probability of an employee being absent from work following an influenza vaccination would be reduced from 5.7 to 1.8% when the incidence of influenza in the community is 6%. Accordingly, if all 1000 employees were vaccinated, a business would be expected to reduce absenteeism from work attributable to an influenza outbreak by 220 days. Moreover, the expected return on every pound invested by an employer would be UK pounds 1.03, UK pounds 3.09 and UK pounds 5.15 (2000 values) when the annual incidence of influenza in the community is 2, 6 and 10%, respectively. CONCLUSIONS: Implementation of an occupational vaccination programme with Influvac would be expected to reduce the incidence of influenza among a workforce leading to less absenteeism from work and averted lost productivity. Even if the incidence of influenza was as low as 2% it may be a worthwhile investment for UK employers to vaccinate their employees with Influvac. PMID- 12093304 TI - Cost effectiveness of paclitaxel/cisplatin compared with cyclophosphamide/cisplatin in the treatment of advanced ovarian cancer in Belgium. AB - OBJECTIVE: To assess the economic impact of two polychemotherapy regimens for patients with advanced ovarian cancer from the perspective of the Belgian health insurance and financing system. DESIGN: An economic evaluation was integrated in an intergroup randomised controlled trial (EORTC 55931) in which patients were randomised to receive the new treatment of paclitaxel and cisplatin or the standard therapy of cyclophosphamide and cisplatin. Data on the use of medical resources were collected prospectively for the 231 European Organization for Research and Treatment of Cancer (EORTC) patients in the trial and costs were valued by using unit prices. The outcome for the economic evaluation was mean survival time as determined by the so-called restricted means method, with the time point of restriction fixed by statistical criteria. A correction of censoring of the cost data collected in the trial was also performed. MAIN OUTCOME MEASURES AND RESULTS: The paclitaxel and cisplatin group experienced a statistically significant improvement in mean survival time of 4 months, which was associated with an increase in the average total cost per patient of 6795 euros (EUR; 1998 values), when costs were assessed over the same period as the gain in mean survival time. This corresponds to a point estimate of the incremental cost-effectiveness ratio of EUR20 385 per life-year gained. The impact of uncertainty was assessed by using a bias-corrected and accelerated bootstrap method with 5000 resamples, and the final results of the analysis are expressed in terms of a cost-effectiveness acceptability curve. CONCLUSIONS: The present economic evaluation has shown that the substitution of paclitaxel for cyclophosphamide in the chemotherapy regimen for women with advanced ovarian cancer leads to a significant improvement in patient survival, which is associated with an increase in costs for the Belgian health insurance system. PMID- 12093306 TI - Therapeutic ineffectiveness: heads or tails? AB - Reporting of therapeutic ineffectiveness through adverse drug reaction spontaneous reporting systems has been proposed by some authors. In the WHO Adverse Reactions Terminology (WHO-ART) and Medical Dictionary for Regulatory Activities (MedDRA) terminology, the term therapeutic ineffectiveness includes drug interactions, resistance, tolerance and tachyphylaxis, as well as pharmaceutical defects such as substandard, adulterated, and counterfeit drugs. Under certain circumstances, reporting therapeutic ineffectiveness may contribute to identifying pharmaceutical defects. However, the best approach to avoid their occurrence would be implementing good manufacturing practices and strengthening quality control activities. This would prevent the misuse of spontaneous reporting of adverse drug reactions (e.g. when reporting of therapeutic ineffectiveness is 'suggested' by interested parties, especially when a generic product has been substituted for a branded original product). PMID- 12093305 TI - 'Sleep attacks' or 'unintended sleep episodes' occur with dopamine agonists: is this a class effect? AB - Controversial reports of sudden onset 'sleep attacks' resulting in road traffic accidents have recently been reported in patients with Parkinson's disease (PD) taking the non-ergot dopamine D(2 )/D(3) receptor agonists pramipexole and ropinirole. These reports have generated considerable debate as the concept of 'sleep attacks' is disputed amongst sleep specialists and most believe that isolated 'sleep attacks' not preceded by warning on the background of chronic sleepiness or 'unintended sleepiness' do not exist. A series of case reports suggested that this phenomenon may not be exclusive to the non-ergot dopamine agonists such as pramipexole or ropinirole and indeed may occur with most dopaminergic agents. Recent evidence suggest that a 'sleepiness' or 'hypoactivity' reaction to dopaminergic therapy may be related to underlying dopamine deficiency of PD rather than a drug effect. In this report we provide the evidence for the phenomenon being a class effect attributable to all dopamine agonists currently employed in the management of PD. Controversy surrounding excessive daytime sleepiness (EDS) in PD and the use of the Epworth Sleepiness Scale (ESS) in relation to PD is also discussed. In spite of variable reports, EDS is recognised to be common in PD and is likely to be related to both the disease process and drug therapy. Studies using multiple sleep latency tests have also reported differing results in PD although a recent study indicated that a subset of 'sleepy' patients with PD may experience pathological somnolence with resultant detrimental consequence on daytime and cognitive functions. We recommend that the issue of 'sleepiness' or 'sleep attacks' in PD should be routinely checked in all patients with PD and indirectly assessed by using either the ESS or the recently introduced Parkinson's Disease Sleep Scale. Those with reported 'sleep attacks' or 'unintended sleep episodes' and excessive daytime sleepiness while taking dopamine agonists or dopaminergic agents such as levodopa should have a review of their medication, should not be driving a car on their own and some may merit formal sleep architecture studies. The latter may identify sleep disorders such as secondary narcolepsy which may benefit from the use of a wakefulness promoting agent. PMID- 12093307 TI - Interrupting drug therapy in the perioperative period. AB - Millions of patients undergo surgery each year and an increasing proportion of these patients are consuming therapeutic drugs. Drug therapy is often withheld in the immediate perioperative period and after major surgery, in particular, there is often a prolonged period of fasting. This may lead to withdrawal effects including recurrence or worsening of patients' disease symptomatology. These effects will occur during a period of physiological and pathophysiological stresses and render patients more vulnerable to drug withdrawal phenomena. Thus, patients may be exposed to greater and sometimes unnecessary risks in the perioperative period. There are relatively few studies that have investigated this problem. The ones that have, however, confirm that drug abstinence in the perioperative period is a relatively common phenomenon and one study has demonstrated an association between duration of drug abstinence and adverse outcomes. The pathophysiological effects of major surgery on gastrointestinal function, neuro-humoral and cytokine adaptive responses to surgical stress are under-appreciated. These responses can reduce the effectiveness of oral administration and exacerbate co-existing disease processes. These problems are compounded by a fragmented approach to perioperative drug therapy with no one group of healthcare professionals assuming responsibility for this aspect of care. This may in part be a consequence of the complexities of rationalising drug therapy in the perioperative period together with the lack of readily available and evidence based information strategies for individual drugs or drug classes. An additional problem relates to the formulations, inherent pharmacokinetics and limited routes of administration of many prescribed drugs. These can prevent a 'seamless' transition from preoperative to postoperative management. Consumers, health professionals, pharmaceutical companies and drug regulatory agencies must all play a part in rectifying this problem. There remains a need for further research to clarify the effects of abstinence on patient outcomes and also to identify optimum strategies to avoid unwanted drug abstinence. PMID- 12093308 TI - A preliminary benefit-risk assessment of lamivudine for the treatment of chronic hepatitis B virus infection. AB - Chronic hepatitis B virus (HBV) infection remains a major public health problem worldwide. Until recently, interferon (IFN)-alpha was the only approved drug for the treatment of chronic HBV infection. The recent registration of lamivudine, a dideoxycytidine analogue that inhibits both the HIV and HBV reverse transcriptases, has provided new perspectives for the treatment of chronic HBV infection. Lamivudine treatment for 12 months leads to a control of viral replication during therapy in the majority of the patients and to sustained anti hepatitis B e (anti-HBe) seroconversion in 16 to 22% of the patients, associated with a biochemical and histological response. Further studies showed that extended lamivudine therapy increases the rate of anti-HBe seroconversion. However, long-term therapy is associated with the progressive emergence of drug resistant mutants. In most cases these mutants are not associated with a deterioration of the liver disease within the available follow-up. In the remaining patients and in particular settings such as liver transplantation, a severe exacerbation of the liver disease is observed and that requires add-on therapy. Lamivudine treatment of patients infected with a pre-core mutant also showed beneficial effect with the control of viral replication, and a biochemical and histological response in approximately 60% of the patients at 1 year. These patients face the same problem of drug resistant mutants, and the optimal duration of lamivudine treatment still needs to be determined in this clinical situation. Moreover, lamivudine therapy is the only therapeutic option in decompensated cirrhotic patients to allow liver transplantation, and in liver transplant patients with HBV recurrence following transplantation. Adverse effects of lamivudine therapy are comparable to those observed in placebo-treated patients. ALT flares have been observed mainly in relation to the re-occurrence of viral replication due to the rebound of viral replication after therapy withdrawal, or to the emergence of drug resistance mutants. Therefore, lamivudine provides a new treatment alternative for patients with chronic HBV infection. For each patient, its indication has to be weighed against the risk of developing viral resistance but also against the risk of natural history of the disease. PMID- 12093310 TI - Management options for cancer therapy-related anaemia. AB - Anaemia is common in patients with haematological malignancy, occurring in the majority of patients with malignant disease who are treated with chemotherapy. Most patients will have their anaemia attributed to the cytokine-mediated anaemia of chronic disease. Many of these patients with anaemia will be symptomatic with fatigue, which is the single most important symptom reported. Data from many studies indicate that treatment of patients with anaemia with recombinant human erythropoietin (rHuEpo) will increase their haemoglobin level, decrease transfusion need and also improve their quality of life. Recent clinical and experimental work suggest that improving the haemoglobin level may improve the patients' prognosis but this finding needs to be confirmed. Treatment of anaemia with rHuEpo in patients with cancer may produce many benefits. Unfortunately, rHuEpo is effective in only around 60% of patients, is slow acting and is expensive. These drawbacks have restricted its use in many healthcare systems. However, a failure to treat anaemia may have important adverse effects for the patient both in terms of their quality of life and, just possibly, in terms of their life expectancy. PMID- 12093309 TI - Benefit-risk considerations in the treatment of dementia with Lewy bodies. AB - Dementia with Lewy bodies (DLB) is a relatively recently characterised syndrome with clinical and pathological features that distinguish it from classical Alzheimer's disease. These characteristics include more rapid decline, spontaneous features of parkinsonism, visual hallucinations and fluctuating cognition. This article reviews the clinical syndrome of DLB and the agents used to treat its cognitive, motor and behavioural manifestations. Benefit-risk issues regarding the treatment of DLB are discussed based upon limited randomised, controlled clinical trials with some speculative conclusions being drawn from case reports and case series. We conclude that patients with DLB may respond better to cholinesterase inhibitors than patients with Alzheimer's disease on both cognitive and behavioural measures. Cholinesterase inhibitor therapy may result in reduced caregiver burden and less time institutionalised. These agents are well tolerated with the majority of adverse effects being gastrointestinal in nature. Although neuropsychiatric manifestations are numerous in patients with DLB, antipsychotics should be used infrequently and with caution, although atypical antipsychotics are better tolerated than conventional antipsychotics. Physicians should exhibit caution when prescribing these agents because of the increased risk of extrapyramidal adverse effects. Limited data suggest that the use of levodopa or other dopaminergic agents may be of benefit for the treatment of the parkinsonism that is associated with DLB. However, the increased risk of hallucinations and neuropsychiatric symptoms may negate the potential benefits of increased mobility. There is insufficient evidence to draw conclusions about the use of antidepressants; however, selective serotonin reuptake inhibitors may be of benefit. PMID- 12093311 TI - Renal failure associated with the use of celecoxib and rofecoxib. AB - OBJECTIVE: Celecoxib and rofecoxib are two relatively new nonsteroidal anti inflammatory drugs (NSAIDs) that selectively inhibit the cyclo-oxygenase-2 (COX 2) isoenzyme at therapeutic concentrations. The nephrotoxic potential of selective COX-2 inhibitors has not been clearly established. This study was conducted in order to understand the association between acute renal failure and the two COX-2 inhibitors celecoxib and rofecoxib. METHODS: A search was performed in the US Food and Drug Administration's (FDA) Adverse Event Reporting System (AERS) to identify cases of renal failure submitted to the FDA. A MEDLINE search of the English language literature was also performed to identify published cases of renal failure associated with celecoxib and rofecoxib. RESULTS: One hundred twenty-two and 142 domestic US cases of celecoxib and rofecoxib-associated renal failure, respectively, were identified in the AERS database. The literature search identified 19 cases of acute renal impairment in association with celecoxib and rofecoxib. In addition, drug regulatory authorities in the UK, Canada, and Australia have received about 50 reports of renal failure with celecoxib and rofecoxib. Descriptive statistics of the AERS cases have been summarised in this report. CONCLUSIONS: Data from AERS and published case reports suggest that use of both these drugs is associated with renal effects similar to that of conventional nonselective NSAIDs. Physicians should be aware that serious or life-threatening renal failure has been reported in patients with normal or impaired renal function after short-term therapy with celecoxib and rofecoxib. Patients at greatest risk for renal injury are those with pre-existing renal impairment, heart failure, liver dysfunction, those taking diuretics and/or ACE inhibitors, and the elderly. Kidney function should be monitored closely for any signs of potential renal injuries soon after initiating treatment with these agents, especially in high-risk populations. In addition, healthcare practitioners should adequately warn patients of the signs and symptoms of serious renal toxicity, and of the need for them to see their physician promptly if they occur. Celecoxib and rofecoxib are not recommended for use in patients with advanced renal disease. PMID- 12093313 TI - Prevention of otitis media by vaccination. AB - Otitis media (OM) is one of the commonest infections in childhood and a frequent reason for prescribing antibacterials in infancy. However, the increase in prevalence of antibacterial-resistant respiratory bacterial pathogens has not been matched by the development of new antibacterial agents. Bacterial vaccine strategies aim to prevent OM directly and to reduce nasopharyngeal carriage of pneumococci, thereby reducing the likelihood of developing acute OM. Complete protection against OM would require an approach targeting both bacterial and viral agents. Immunisation with a pneumococcal conjugate vaccine provides protection against acute OM caused by pneumococcal serotypes included in the vaccine, reduces serotype-specific pneumococcal carriage, and reduces carriage of penicillin-resistant pneumococci. However, an increase in non-vaccine serotype OM has been observed in vaccinated children, which may limit the overall effectiveness of this vaccine. New vaccines targeting non-typable Haemophilus influenzae and Mycoplasma catarrhalis are in the early stages of development. Efficacy studies with influenza vaccine have shown the most promising results to date in terms of overall reduction in OM episodes. A more substantial reduction in the burden of OM in childhood would require a combination of vaccines that are effective against the bacterial and viral pathogens involved and that can be administered early in infancy. PMID- 12093312 TI - Angiotensin II receptor antagonists in chronic heart failure: where do they fit? AB - Heart failure is a common and disabling condition with a dismal prognosis. Inhibition of the renin-angiotensin-aldosterone system (RAAS) with angiotensin converting enzyme (ACE) inhibitors has proven to be a valuable therapeutic strategy in this condition, with well-proven morbidity and mortality benefits. Nonetheless, ACE inhibitors provide incomplete blockade of the RAAS and also inhibit the degradation of bradykinin. Although increased levels of bradykinin may have haemodynamic advantages by contributing to vasodilatation, they may also be largely responsible for some of the adverse effects of ACE inhibitors. Angiotensin II (Ang II) receptor antagonists offer more complete blockade of the RAAS without the potentiation of bradykinin, and it was therefore hoped that they would provide even greater benefits for patients with heart failure. So far, much of the initial promise of the Ang II receptor antagonists in heart failure has not been realised. There has been no conclusive demonstration of their superiority to ACE inhibitors in their effects on morbidity and mortality, and their equivalence to ACE inhibitors has not been proven. The Ang II receptor antagonists have, however, proven to be better tolerated than ACE inhibitors and they are therefore likely to be a reasonable alternative for those patients with heart failure who cannot tolerate ACE inhibition. Recent evidence has indicated that the Ang II type 1 receptor antagonist valsartan is of value when used in patients already receiving either an ACE inhibitor or a beta-blocker, but has also suggested that giving all three drugs together is deleterious. Further evidence about the value of Ang II receptor antagonists in heart failure may be provided by further studies, of which several are currently ongoing. PMID- 12093314 TI - Management issues in syphilis. AB - Syphilis is a sexually transmitted infection which is systemic from the outset and has increased in incidence worldwide over the last decade. There has been concern as to whether or not co-infection with HIV can modify the clinical presentation of syphilis and, as a genital ulcer disease, it can facilitate the transmission of HIV infection. Diagnosis is based on the microscopic identification of the causative treponeme and serological testing. Recommendations for the treatment of syphilis have been based on expert opinion, case series, some clinical trials and 50 years of clinical experience. Penicillin, given intramuscularly, is the mainstay of treatment and the favoured preparations for early infectious syphilis are benzathine penicillin as a single injection or a course of daily procaine penicillin injections for 10 to 14 days. The duration of treatment is longer for late syphilis. There has been concern that benzathine penicillin may not prevent the development of neurosyphilis but that is a rare outcome with this therapy. The main alternative to penicillin is doxycycline, but the place of azithromycin and ceftriaxone is yet to be established. It is not necessary to carry out examination of the cerebrospinal fluid in patients with early infectious syphilis but it should be performed in those with neurological or ocular signs, psychiatric signs or symptoms, when there is evidence of treatment failure and in those who are co-infected with HIV. Follow-up is an essential part of management and should be particularly assiduous, for at least 24 months, in those co-infected with HIV. Partner notification should be mandatory to try to contain the spread of infection. PMID- 12093315 TI - Metabolic and additional vascular effects of thiazolidinediones. AB - Several cardiovascular risk factors (dyslipidaemia, hypertension, glucose intolerance, hypercoagulability, obesity, hyperinsulinaemia and low-grade inflammation) cluster in the insulin resistance syndrome. Treatment of these individual risk factors reduces cardiovascular complications. However, targeting the underlying pathophysiological mechanisms of the insulin resistance syndrome is a more rational treatment strategy to further improve cardiovascular outcome. Our understanding of the so-called cardiovascular dysmetabolic syndrome has been improved by the discovery of nuclear peroxisome proliferator-activated receptors (PPARs). PPARs are ligand-activated transcription factors belonging to the nuclear receptor superfamily. As transcription factors, PPARs regulate the expression of numerous genes and affect glycaemic control, lipid metabolism, vascular tone and inflammation. Activation of the subtype PPAR-gamma improves insulin sensitivity. Expression of PPAR-gamma is present in several cell types involved in the process of atherosclerosis. Thus, modulation of PPAR-gamma activity is an interesting therapeutic approach to reduce cardiovascular events. Thiazolidinediones are PPAR-gamma agonists and constitute a new class of pharmacological agents for the treatment of type 2 (non-insulin-dependent) diabetes mellitus. Two such compounds are currently available for clinical use: rosiglitazone and pioglitazone. Thiazolidinediones improve insulin sensitivity and glycaemic control in patients with type 2 diabetes. In addition, improvement in endothelial function, a decrease in inflammatory conditions, a decrease in plasma levels of free fatty acids and lower blood pressure have been observed, which may have important beneficial effects on the vasculature. Several questions remain to be answered about PPAR-gamma agonists, particularly with respect to the role of PPAR-gamma in vascular pathophysiology. More needs to be known about the adverse effects of thiazolidinediones, such as hepatotoxicity, increased low density lipoprotein cholesterol levels and increased oedema. The paradox of adipocyte differentiation with weight gain concurring with the insulin sensitising effect of thiazolidinediones is not completely understood. The decrease in blood pressure induced by thiazolidinedione treatment seems incompatible with an increase in the plasma volume, and the discrepancy between the stimulation of the expression of CD36 and the antiatherogenic effects of the thiazolidinediones also needs further explanation. Long-term clinical trials of thiazolidinediones with cardiovascular endpoints are currently in progress. In conclusion, studying the effects of thiazolidinediones may shed more light on the mechanisms involved in the insulin resistance syndrome. Furthermore, thiazolidinediones could have specific, direct effects on processes involved in the development of vascular abnormalities. PMID- 12093317 TI - Esomeprazole: a review of its use in the management of acid-related disorders. AB - Esomeprazole (S-isomer of omeprazole), the first single optical isomer proton pump inhibitor, generally provides better acid control than current racemic proton pump inhibitors and has a favourable pharmacokinetic profile relative to omeprazole. In a large well designed 8-week trial in patients (n >5000) with erosive oesophagitis, esomeprazole recipients achieved significantly higher rates of endoscopically confirmed healed oesophagitis than those receiving lansoprazole. Respective healed oesophagitis rates with once-daily esomeprazole 40mg or lansoprazole 30mg were 92.6 and 88.8%. Overall, esomeprazole was also better than omeprazole, although these differences were not always statistically significance. Ninety-two to 94% of esomeprazole recipients (40mg once daily) achieved healed oesophagitis versus 84 to 90% of omeprazole recipients (20mg once daily). Esomeprazole was effective across all baseline grades of oesophagitis; notably, relative to lansoprazole, as baseline severity of disease increased, the difference in rates of healed oesophagitis also increased in favour of esomeprazole. Resolution of heartburn was also significantly better with esomeprazole 40mg than with these racemic proton pump inhibitors. Long-term (up to 12 months) therapy with esomeprazole effectively maintained healing in these patients. Once-daily esomeprazole 20 or 40mg for 4 weeks resolved symptoms in patients with symptomatic gastro-oesophageal reflux disease (GORD) without oesophagitis. Symptoms were effectively managed in the long-term with symptom driven on-demand esomeprazole (20 or 40mg once daily). Eradicating Helicobacter pylori infection is considered pivotal to successfully managing duodenal ulcer disease. Seven days' treatment (twice-daily esomeprazole 20mg plus amoxicillin 1g and clarithromycin 500mg) eradicated H. pylori in >/=86% of patients (intention to-treat), a rate that was similar to equivalent omeprazole-based regimens. Esomeprazole is generally well tolerated, both as monotherapy and in combination with antimicrobial agents, with a tolerability profile similar to that of other proton pump inhibitors. Few patients discontinued therapy because of treatment emergent adverse events (<3% of patients), with very few (<1%) drug-related serious adverse events reported. CONCLUSIONS: Esomeprazole is an effective, well tolerated treatment for managing GORD and for eradicating H. pylori infection in patients with duodenal ulcer disease. In 8-week double-blind trials, esomeprazole healed oesophagitis and resolved symptoms in patients with endoscopically confirmed erosive oesophagitis and overall, provided better efficacy than omeprazole. Notably, in a large (n >5000 patients) double-blind trial, esomeprazole 40mg provided significantly better efficacy than lansoprazole in terms of healing rates and resolution of symptoms. Long-term therapy with esomeprazole maintained healed oesophagitis in these patients. Esomeprazole also proved beneficial in patients with symptomatic GORD without oesophagitis. Thus, esomeprazole has emerged as an option for first-line therapy in the management of acid-related disorders. PMID- 12093319 TI - Ethical challenges in end-of-life therapies in the elderly. AB - With the increasing numbers of elderly in the population of all western countries and the increasing life expectancy at birth, many seniors spend the last period of their life with various afflictions that may require the need for long-term institutional care. During the last period of life, many seniors and their families face decisions that challenge ethical principles and may cause conflict among family members as well as healthcare professionals. The commonly used ethical principles of autonomy, beneficence, nonmaleficence and justice, although forming a useful foundation for the evaluation of decision-making dilemmas, alone cannot resolve many clinically challenging situations. Healthcare professionals must clearly understand the clinical state of the patient for whom a difficult decision is being contemplated. Levels of function, clinical symptoms, the expected trajectory of change and possible treatment options have to be balanced against the person's values and wishes, either self-expressed directly or through an advance directive, or communicated by surrogate decision makers. At times, physicians face difficult treatment dilemmas when patients or families request treatments that are not legally sanctioned, such as when physician-assisted suicide is requested by a suffering patient. At other times conflicts occur when patients or surrogates wish to continue with therapies that are no longer considered necessary or suitable by the physician. At the societal level, sometimes an expensive drug that is deemed necessary by the physicians is not covered by a government-sponsored or private health plan. The issue of distributive justice must be considered in a situation such as when long-term facilities or acute hospitals treating frail, cognitively impaired elders consider withholding or withdrawing various treatments because of poor clinical outcomes coupled with excessive costs. The often controversial issue of nutrition and hydration in the end-of-life period frequently causes treatment conflicts and dilemmas among surrogates and staff, as does the highly charged issue of cardiopulmonary resuscitation in this frail and very vulnerable population. The real challenge for healthcare providers in the field of geriatric long-term care is to balance compassionate and appropriate care with respect for the choices and wishes of patients and their families. This should be accomplished while at the same time safeguarding the professional standards and ethical integrity of healthcare providers responsible for this care. PMID- 12093320 TI - Bacterial skin infections in the elderly: diagnosis and treatment. AB - Skin and soft tissue infections are quite common in elderly people. A number of special conditions and circumstances need to be considered in the diagnosis and therapy. It is important to try to establish the causative organism, exclude other cutaneous disorders and identify precipitating factors. Treatment modalities include antiseptics, topical and systemic antibacterials, dressings and biotherapy. Skin infections presenting with erythema, blisters, pustules, and ulcerations or in body folds are described in detail. Cellulitis and infected ulcers are the most commonly encountered cutaneous infections in the elderly. Accurate and quick diagnosis and treatment are imperative to prevent significant morbidity and mortality. Appropriate antibacterials, antiseptics and dressings are necessary depending on the severity of the clinical presentation and resistance patterns. Laboratory tests, such as skin swabs, to establish the exact pathogen take time and the results might represent colonisation rather than infection of the skin. Cellulitis should be clinically distinguished from erysipelas and necrotising fasciitis. The latter is a life-threatening condition, which in the majority of cases requires surgical debridement of the infected tissue. Blisters and honey-coloured crusts are typical features of impetigo. It is very contagious and close contacts should be examined. Folliculitis is a commonly seen skin infection, which often responds to the use of antiseptics and topical antibacterials. More severe pustular skin eruptions, such as furunculosis and carbunculosis, usually require treatment with systemic antibacterials. Intertrigo and erythrasma have a predilection for the body folds, especially the axillae and groin, and topical therapy is usually sufficient. Secondary skin infections are often the result of persistent pruritus associated with increasing dryness of the aging skin. Emollients and antihistamines are useful measures. Primary cutaneous disorders and systemic diseases should be excluded with the aid of appropriate investigations, such as blood tests and skin biopsy. Staphylococcus aureus and beta-haemolytic streptococci are the most common causative organisms of cutaneous infections. PMID- 12093318 TI - Rizatriptan: an update of its use in the management of migraine. AB - Rizatriptan is an orally active serotonin 5-HT(1) receptor agonist that potently and selectively binds to 5-HT(1B/1D) subtypes. Earlier clinical trials demonstrated that rizatriptan 5 or 10mg is more effective than placebo at providing pain relief and a pain-free state, relieving associated symptoms of migraine, normalising functional ability and improving patient quality of life, and showed that rizatriptan provides faster freedom from pain and reduces nausea to a greater extent than oral sumatriptan. More recently, rizatriptan 10mg was shown to be more effective than zolmitriptan 2.5mg or naratriptan 2.5mg at producing a pain-free state 2 hours postdose. Furthermore, compared with naratriptan, significantly more patients who received rizatriptan were pain free or had pain relief from 1 hour onwards. The number of patients with normal functional ability at 2 hours was significantly higher after rizatriptan than after naratriptan or zolmitriptan. Rizatriptan was also generally more effective than zolmitriptan or naratriptan at relieving migraine-associated symptoms. Rizatriptan is generally well tolerated and adverse events are usually mild and transient. The most common adverse events associated with rizatriptan in recent randomised trials were asthenia/fatigue, dizziness, somnolence and nausea. There was a trend towards a lower incidence of adverse events with rizatriptan compared with zolmitriptan (31.2 vs 38.8%). However, rizatriptan was associated with a significantly higher incidence of adverse events than naratriptan (39 vs 29%). The incidence of chest pain was similar after the administration of rizatriptan, zolmitriptan or naratriptan (2 to 4%). CONCLUSION: Rizatriptan is an effective drug for the acute treatment of moderate or severe migraine. Oral rizatriptan 5 and 10mg have shown greater efficacy than placebo in providing pain relief, an absence of pain, relief from associated symptoms, normal functional ability and an improvement in patient quality of life. Earlier results showed that rizatriptan provides faster freedom from pain and reduces nausea to a greater extent than oral sumatriptan. More recent studies have shown that rizatriptan 10mg provides faster pain relief and a higher percentage of patients with an absence of pain and normal functional ability at 2 hours than naratriptan 2.5mg or zolmitriptan 2.5mg. The efficacy of rizatriptan is retained when used in the long term and the drug is generally well tolerated. Although well designed studies comparing rizatriptan with almotriptan, eletriptan and frovatriptan would further define the position of rizatriptan, current data suggest rizatriptan should be considered as a first-line treatment option in the management of migraine. PMID- 12093321 TI - Prophylaxis of herpesvirus infections in immunocompetent and immunocompromised older patients. AB - In older patients, prophylaxis of herpesvirus infections mainly involves preventing the recurrence of herpes simplex virus (HSV) and complications of herpes zoster in immunocompetent patients, while in immunocompromised patients it is more concerned with the prevention of opportunistic virus reactivation. HSV ocular infection is the most frequent cause of corneal blindness in the US. The effectiveness of aciclovir 400mg twice daily in preventing the recurrence of HSV eye disease in immunocompetent patients has been well demonstrated. The issue of treatment duration for patients with highly recurrent ocular herpes remains unresolved. Post-herpetic neuralgia (PHN) is one of the most common neuralgic illnesses worldwide. Some progress in prevention of PHN has been made with a combination of antiviral therapy (famciclovir or valaciclovir), started within 72 hours of onset of the rash, and analgesic treatment. However, the best prevention of PHN is the prevention of herpes zoster disease, and the varicella vaccine is an option which over the next few years will be tested in clinical trials. For immunocompromised patients of any age, restoring immunity prevents herpesvirus disease, as demonstrated for cytomegalovirus (CMV) in AIDS patients receiving highly active antiretroviral therapy. Specific antiviral therapy during the initial period after transplantation could prevent reactivation of HSV or CMV in seropositive recipients. Whether pre-emptive therapy or prophylaxis with ganciclovir is the optimal approach against CMV remains controversial, and the relative merits and limitations of each approach may guide the choice. In stem cell transplantation, pre-emptive therapy with foscarnet avoids the neutropenia and related complications associated with ganciclovir. In renal transplant recipients, universal prophylaxis of CMV infection with valaciclovir has the same efficacy as ganciclovir. Although it is relatively toxic, cidofovir should be further evaluated because of its in vitro activity against most DNA viruses. PMID- 12093322 TI - Treatment of inflammatory bowel disease in the elderly: an update. AB - Inflammatory bowel disease (IBD) is most common in young adults, but it can also present in the elderly. Furthermore, with the aging of the population, the number of elderly patients with IBD is expected to grow. Other conditions, such as diverticulitis and ischaemic colitis, may be more common in the elderly and need to be considered in the differential diagnosis. Management of elderly patients with IBD follows the same principles as in younger patients, with a few exceptions. For patients with mild-to-moderate colitis, a 5-aminosalicylate drug is often used (sulfasalazine, olsalazine, mesalazine, balsalazide). Topical therapy may be sufficient for those with distal colitis, whereas an oral preparation is used for more extensive disease. In those with more severe or refractory symptoms, corticosteroids are used, although the elderly appear to be at increased risk for corticosteroid-associated complications. For patients with corticosteroid-dependent or corticosteroid-refractory disease, immunosuppression with azathioprine or mercaptopurine may help avoid surgery. In patients with Crohn's disease, a similar approach is followed, with the additional consideration that the formulation of drug used must ensure delivery of drug to the site of inflammation. In fistulising Crohn's disease, antibacterials, immunosuppressive drugs, infliximab and surgery are often used in combination. Controlled trials and clinical experience have shown that infliximab is a significant addition to the therapeutic armamentarium for patients with Crohn's disease. PMID- 12093316 TI - Food-drug interactions. AB - Interactions between food and drugs may inadvertently reduce or increase the drug effect. The majority of clinically relevant food-drug interactions are caused by food-induced changes in the bioavailability of the drug. Since the bioavailability and clinical effect of most drugs are correlated, the bioavailability is an important pharmacokinetic effect parameter. However, in order to evaluate the clinical relevance of a food-drug interaction, the impact of food intake on the clinical effect of the drug has to be quantified as well. As a result of quality review in healthcare systems, healthcare providers are increasingly required to develop methods for identifying and preventing adverse food-drug interactions. In this review of original literature, we have tried to provide both pharmacokinetic and clinical effect parameters of clinically relevant food-drug interactions. The most important interactions are those associated with a high risk of treatment failure arising from a significantly reduced bioavailability in the fed state. Such interactions are frequently caused by chelation with components in food (as occurs with alendronic acid, clodronic acid, didanosine, etidronic acid, penicillamine and tetracycline) or dairy products (ciprofloxacin and norfloxacin), or by other direct interactions between the drug and certain food components (avitriptan, indinavir, itraconazole solution, levodopa, melphalan, mercaptopurine and perindopril). In addition, the physiological response to food intake, in particular gastric acid secretion, may reduce the bioavailability of certain drugs (ampicillin, azithromycin capsules, didanosine, erythromycin stearate or enteric coated, and isoniazid). For other drugs, concomitant food intake may result in an increase in drug bioavailability either because of a food-induced increase in drug solubility (albendazole, atovaquone, griseofulvin, isotretinoin, lovastatin, mefloquine, saquinavir and tacrolimus) or because of the secretion of gastric acid (itraconazole capsules) or bile (griseofulvin and halofantrine) in response to food intake. For most drugs, such an increase results in a desired increase in drug effect, but in others it may result in serious toxicity (halofantrine). PMID- 12093326 TI - Pancreatoduodenectomy for pancreatic adenocarcinoma: triumph, triumphalism, or transition? PMID- 12093328 TI - Perianal Crohn disease: a new scoring system to evaluate and predict outcome of surgical intervention. AB - BACKGROUND: Conventional Crohn disease activity indices do not reflect perianal disease activity or allow prognostic implications from surgery. HYPOTHESIS: A new scoring system, based on the patient's disease history and physical examination findings, will allow more accurate use of surgical intervention for perianal Crohn disease. METHODS: A standardized scoring questionnaire was developed and applied to a consecutive group of patients before surgical treatment of perianal Crohn disease. The scoring system included abscess, fistula, ulcer and fissure, stenosis, incontinence, and concomitant disease. Weighted factors included acuity vs chronicity, de novo vs recurrent disease, and concomitant intestinal disease. The scoring system was validated against the surgical outcome, which was classified as poor, satisfactory, or good. RESULTS: Twenty-eight patients with Crohn disease who underwent 33 surgical procedures had a mean score of 16.5 (range, 3-37; possible range, 0-55). Using the Spearman nonparametric correlation test, the scoring system was accurate in predicting the outcome of surgical intervention (correlation coefficient, 0.78, 95% confidence interval, 0.57-0.89; P<.001) at mean follow-up of 20.8 months (range, 6-40 months). Correlation was further validated using a linear regression model (r = 0.75, slope best-fit value, 3.8; 95% confidence interval, 2.46-5.14; P<.001). All patients with a score of 10 or less had a good outcome, whereas all those with a score of 20 or greater had a poor outcome. CONCLUSIONS: The proposed scoring system correlated well with the short-term outcome of surgical intervention in patients with perianal Crohn disease and allowed prediction of surgical success. Ultimately, it may be possible to alter therapy based on preoperative prediction of the expected postoperative outcome. PMID- 12093324 TI - Sertraline: a review of its use in the management of major depressive disorder in elderly patients. AB - Sertraline is a selective serotonin reuptake inhibitor (SSRI) with well established antidepressant and anxiolytic activity. Results from several well designed trials show that sertraline (50 to 200 mg/day) is effective in the treatment of major depressive disorder in elderly patients (> or =60 years of age). Primary endpoints in most studies included the Hamilton Depression Rating Scale (HDRS), Clinical Global Impression (CGI) score and the Montgomery-Asberg Depression Rating Scale (MADRS). Sertraline was significantly more effective than placebo, and was as effective as fluoxetine, nortriptyline and imipramine in elderly patients. During one trial, amitriptyline was significantly more effective than sertraline [mean reduction from baseline on one of six primary outcomes (HDRS)], although no quantitative data were provided. Subgroup analysis of data from a randomised, double-blind trial in elderly patients with major depressive disorder suggests that vascular morbidity, diabetes mellitus or arthritis does not affect the antidepressant effect of sertraline. Secondary endpoints from these clinical trials suggest that sertraline has significant benefits over nortriptyline in terms of quality of life. In addition, significant differences favouring sertraline in comparison with nortriptyline and fluoxetine have been recorded for a number of cognitive functioning parameters. Sertraline is generally well tolerated in elderly patients with major depressive disorder, and lacks the marked anticholinergic effects that characterise the adverse event profiles of tricyclic antidepressants (TCAs). The most frequently reported adverse events in patients aged > or =60 years with major depressive disorder receiving sertraline 50 to 150 mg/day were dry mouth, headache, diarrhoea, nausea, insomnia, somnolence, constipation, dizziness, sweating and taste abnormalities. The tolerability profile of sertraline is generally similar in younger and elderly patients. Sertraline has a low potential for drug interactions at the level of the cytochrome P450 enzyme system. In addition, no dosage adjustments are warranted for elderly patients solely based on age. CONCLUSION: Sertraline is an effective and well tolerated antidepressant for the treatment of major depressive disorder in patients aged > or =60 years. Since elderly patients are particularly prone to the anticholinergic effects of TCAs as a class, SSRIs such as sertraline are likely to be a better choice for the treatment of major depressive disorder in this age group. In addition, sertraline may have advantages over the SSRIs paroxetine, fluoxetine and fluvoxamine in elderly patients because of the drug's comparatively low potential for drug interactions, which is of importance in patient groups such as the elderly who are likely to receive more than one drug regimen. PMID- 12093323 TI - Lung cancer in the elderly: current and future chemotherapeutic options. AB - Lung cancer is a prevalent malignancy disproportionately affecting the elderly, and in our aging societies will only increase in magnitude. Physicians typically assume that elderly lung cancer patients will have poorer prognoses. This belief is in part based on certain physiological changes of aging affecting the kidneys, liver, and bone marrow. However, there are no data to clearly support or refute increased toxicity from chemotherapy or a lessened therapeutic effect in the elderly based on these changes, although it is a field worthy of further study. Retrospective studies of treatment of elderly non-small cell and small cell lung cancer patients do not suggest a worse prognosis based on advanced age alone. Clinicians are hampered by the lack of clinical trials focusing on or even including the elderly, despite the increased incidence of lung cancer in the elderly. Phase II studies in elderly non-small cell lung cancer patients concentrate on newer agents (vinorelbine and gemcitabine) alone or combined with platinum compounds in hopes of more favourable toxicity profiles. Phase III trials have demonstrated survival benefits, quality of life improvements, and acceptable toxicity profiles for vinorelbine compared to best supportive care alone and the combination of vinorelbine and gemcitabine compared to vinorelbine alone. Data are also sparse for elderly small cell lung cancer patients. Phase II studies focused on single agent oral etoposide also in hopes of lessening toxicity. However, phase III trials have shown improvement in survival and quality of life for multiagent intravenous chemotherapy compared to oral etoposide. Given the existing data, altering therapy for lung cancer patients based on age alone would not be warranted. Given the prevalence of the disease, future studies need to include an appropriate number of elderly patients with continued emphasis on quality of life in addition to survival. PMID- 12093330 TI - Outcome study of psychological distress and nonspecific symptoms in patients with mild primary hyperparathyroidism. AB - BACKGROUND: Primary hyperparathyroidism is a common endocrinopathy. The appropriate management of its mild form, however, remains controversial. HYPOTHESIS: Mild primary hyperparathyroidism is associated with psychological distress and other nonspecific symptoms that improve following parathyroidectomy. DESIGN: Two-year prospective before-after study. SETTING: University hospital. PATIENTS: Twenty-six consecutive patients with mild hypercalcemia (<12 mg/dL [<3 mmol/L]) due to primary hyperparathyroidism, without osteitis fibrosa cystica or urolithiasis were enrolled from January 11, 1997, through April 21, 1998. INTERVENTION: Parathyroidectomy. MAIN OUTCOME MEASURES: Primary outcome was psychological distress as measured by the 28-item version of the General Health Questionnaire. Secondary outcomes included body weight, joint pain, and occurrences of bowel movements and urination. RESULTS: Before surgery, 15 patients (58%; 95% confidence interval, 37%-77%) showed psychological distress (case group) while 11 patients did not (noncase group). A clinically and statistically significant reduction in the General Health Questionnaire score was detected at 3 months in the case group (-6.1; 95% confidence interval, -11.0 to 1.2), but the reduction was smaller (-1.9; 95% confidence interval, -6.9 to 3.0) at 24 months after surgery. No significant change in the General Health Questionnaire score was observed in the noncase group during the follow-up. No significant change was noted in any of the secondary outcomes. CONCLUSIONS: Psychological distress was associated with mild primary hyperparathyroidism and was ameliorated after surgery. The improvement, however, was limited in extent and duration. PMID- 12093333 TI - Male sex predisposes the newborn surgical patient to parenteral nutrition associated cholestasis and to sepsis. AB - HYPOTHESIS: Sepsis is an epiphenomenon of parenteral nutrition-associated cholestasis (PNAC) and not a causative factor, and the incidence of sepsis is not affected by the presence or absence of PNAC. DESIGN: Observational cohort study. SETTING: Pediatric surgery department in a tertiary referral children's hospital. PATIENTS: Newborns receiving PN for at least 7 days following intestinal surgery. MAIN OUTCOME MEASURES: The criteria for PNAC were as follows: PN for at least 14 consecutive days, conjugated bilirubin level greater than 1.5 mg/dL (>26 micromol/L), conjugated bilirubin fraction greater than 50%, and absence of another identifiable cause of cholestasis. The identification of septic events was based on Centers for Disease Control and Prevention criteria. RESULTS: The patients (26 with PNAC and 72 without PNAC) were well comparable for underlying disease, gestational age, birth weight, and age at the start of PN. Time receiving PN and length of hospital stay were significantly (P<.001) longer in patients with PNAC. Parenteral nutrition-associated cholestasis was associated with male sex (P =.03; odds ratio, 2.8; 95% confidence interval, 1.1-7.1). The overall sepsis incidence was low (9 per 1000 hospital days). The sepsis incidence tended to be higher in patients with PNAC than in patients without PNAC (11.8 vs 7.1 per 1000 days; P =.08), but was significantly higher in male than in female patients (12.2 vs 5.6 per 1000 days; P =.01). Most septic events were caused by coagulase-negative staphylococci. CONCLUSIONS: Sepsis is an epiphenomenon of PNAC rather than a causative factor. Moreover, male sex predisposes the newborn surgical patient to PNAC and to sepsis. PMID- 12093332 TI - Bovine pericardium vs dacron for patch angioplasty after carotid endarterectomy: a prospective randomized study. AB - HYPOTHESIS: Bovine pericardium (BP) demonstrates improved intraoperative hemostasis and equivalent perioperative morbidity compared with Dacron when used as patch material for angioplasty following carotid endarterectomy. OBJECTIVE: To prospectively compare BP and Dacron patch angioplasty after carotid endarterectomy in a randomized fashion. METHODS: Ninety-five consecutive primary carotid endarterectomies were performed in a prospective randomized fashion in 92 patients. Fifty-one procedures were performed using BP and 44 using Dacron. Intraoperative suture line bleeding was subjectively evaluated by observing bleeding at 3 and 4 minutes following carotid cross-clamp removal and then objectively weighing the sponge used to tamponade bleeding during these time intervals. Perioperative morbidity, including cervical wound hematoma, transient ischemic attack, and stroke, and perioperative mortality were recorded. Statistical analysis was performed using paired t tests, chi(2) analysis, Fisher exact test, or multiple linear regression as appropriate. RESULTS: Suture line bleeding at 3 minutes was present in 7 (14%) of 51 patients in the BP group and 24 (55%) of 44 patients in the Dacron group (P<.001). Suture line bleeding evaluated at 4 minutes was present in 2 (4%) of 51 patients in the BP group and 13 (30%) of 44 patients in the Dacron group (P =.001). Net +/- SEM sponge weight (total intraoperative suture line bleeding) was 6.25 +/- 0.55 g in the BP group and 16.34 +/- 1.85 g in the Dacron group (P<.001). Total suture line bleeding was significantly affected by activated clotting time; however, multivariate analysis demonstrated that bleeding was significantly less with BP (P<.001) even after adjusting for differences in activated clotting time. CONCLUSIONS: Bovine pericardium demonstrated a statistically significant decrease in intraoperative suture line bleeding compared with Dacron. Handling characteristics were judged by the surgeons to be superior for BP. Therefore, we believe BP may be an alternative to Dacron when performing patch angioplasty of the carotid artery after endarterectomy. PMID- 12093334 TI - Surgical undergraduate education in rural Australia. AB - HYPOTHESIS: Surgical undergraduate education in a rural setting is feasible and sound in terms of educational outcomes. DESIGN: The final-year surgical curriculum at the University of Adelaide, Adelaide, South Australia, was restructured to include the option of a rural surgical term. SETTING: Five provincial center hospitals in rural South Australia. INTERVENTIONS: Forty-three final-year medical students undertook rural surgical clerkships in 1998. MAIN OUTCOME MEASURES: End-of-year results and subjective ward assessments were compared between the group of students who completed rural surgical terms and the remainder of the student group who participated in tertiary hospital-based electives. Subjective student feedback was obtained in a survey conducted by the Clinical Education Development Unit at the University of Adelaide. RESULTS: No significant (P =.45) differences in examination results were noted between the rural and city groups. A significant (P<.01) finding was observed in the subjective assessments, indicating that it was more difficult for the rural group to obtain an A grade compared with the city group. The rural students ranked the level of teaching and supervision highly and enjoyed the overall rural experience. CONCLUSION: Surgical undergraduate education is practical in a rural setting and, for educational outcome, seems to be at least as effective as city based surgical clerkships in preparing students for final examinations. PMID- 12093335 TI - The clinical and economic correlates of misdiagnosed appendicitis: nationwide analysis. AB - BACKGROUND: Negative appendectomy (NA)--the nonincidental removal of a normal appendix--occurs commonly but the associated clinical- and system-level costs are not well studied. HYPOTHESIS: The frequency of adverse clinical outcomes and associated financial burden of hospitalizations during which NA is performed is greater than previously recognized and varies widely among demographic groups. DESIGN: Population-based, retrospective cohort study. SETTING: The 1997 Nationwide Inpatient Sample of the Health Care Utilization Project. PATIENTS: All surveyed patients assigned International Classification of Diseases, Ninth Revision procedure codes for appendectomy but without an associated diagnosis of acute appendicitis. MAIN OUTCOME MEASURES: The age- and sex-stratified rates of NA, the incidence of associated infectious complications and case fatality, and the average length of stay and hospitalization charges during those admissions. RESULTS: Nationwide, an estimated 261 134 patients underwent nonincidental appendectomies in 1997, and 39 901 (15.3%) were negative for appendicitis. Women had a higher rate of NA as did patients younger than 5 years and older than 60 years. When compared with patients with appendicitis, NA was associated with a significantly longer length of stay (5.8 vs 3.6 days, P<.001), total charge admission ($18 780 vs $10 584, P<.001), case fatality rate (1.5% vs 0.2%, P<.001), and rate of infectious complications (2.6% vs 1.8%, P<.001). An estimated $741.5 million in total hospital charges resulted from admissions in which a NA was performed. CONCLUSIONS: There are significant clinical and financial costs incurred by patients undergoing NA during the treatment of presumed appendicitis. These should be considered when evaluating system-level interventions to improve the management of appendicitis. PMID- 12093337 TI - Cholesterol and serum albumin levels as predictors of cross infection, death, and length of hospital stay. AB - HYPOTHESIS: The levels of cholesterol, its fractions (high-density lipoprotein cholesterol [HDL-C] and low-density lipoprotein cholesterol [LDL-C]), and serum albumin reflect nutritional status and are related to in-hospital death, nosocomial infection, and length of stay in the hospital. DESIGN: A prospective cohort study of hospitalized patients. SETTING: The Service of General Surgery of a tertiary hospital. PATIENTS: A consecutive series of 2989 patients admitted for more than 1 day. MAIN OUTCOME MEASURES: Nosocomial infection, in-hospital death, and length of stay. RESULTS: During follow-up, 62 (2%) of the patients died, 382 (13%) developed a nosocomial infection, and 257 (9%) developed a surgical site infection. Serum albumin (lowest quintile vs highest quintile: adjusted odds ratio [OR], 1.9; 95% confidence interval, 1.2-2.9) and HDL-C (lowest quintile vs highest quintile: OR, 2.0; 95% confidence interval, 1.3-3.0) levels showed an inverse and highly significant relationship with nosocomial infection (mainly due to surgical site infection) in crude and multivariate analyses (controlling for the Study on the Efficacy of Nosocomial Infection Control [SENIC] index, the American Society of Anesthesiologists' score, cancer, and age). Regarding total and LDL-C levels, only their lowest quintiles increased the risk of nosocomial infection. Serum albumin and HDL-C levels showed an inverse trend (P<.001) with mortality, with high multivariate-adjusted ORs in the lowest quintile (serum albumin: OR, 5.8; 95% confidence interval, 0.8-44.6; HDL-C: OR, 7.2; 95% confidence interval, 0.9-55.0), whereas no trend was appreciated with other cholesterol fractions or ratios. Serum albumin, HDL-C, and LDL-C levels showed independent, significant (P<.001), and inverse relationships with length of stay. CONCLUSION: The levels of serum albumin and cholesterol fractions, mainly HDL-C, which are routinely measured at hospital admission, are predictors of in-hospital death, nosocomial infection, and length of stay. PMID- 12093338 TI - Effect of topically applied charged particles on healing of colonic anastomoses. AB - HYPOTHESIS: Various forms of electrical stimulation can improve wound healing in different tissues, but their application to gastrointestinal tract healing has not been investigated. We assumed that positively charged diethylaminoethyl cross linked dextran bead (diethylaminoethyl Sephadex [DEAE-S]) particles would have a beneficial effect on the healing of colonic anastomoses. DESIGN: Experimental animal study. SETTING: Animal research laboratory of a university hospital. ANIMALS: Forty female Wistar albino rats. INTERVENTIONS: Right colonic transection and anastomosis was performed in 5 animal groups. The control group received no treatment; the placebo group, methylcellulose gel; and the DEAE-S group, DEAE-S in methyl cellulose gel applied topically around the anastomoses. The fecal peritonitis (FP) group underwent cecal ligation and perforation simultaneously with the anastomosis to cause FP; the FP + DEAE-S group also received DEAE-S applied around the anastomoses. MAIN OUTCOME MEASURES: After the completion of postoperative day 4, all rats were killed. Anastomotic bursting pressures and hydroxyproline concentrations in perianastomotic tissue were measured and compared. RESULTS: Mean bursting pressures were 115.1 mm Hg in the control group, 113.6 mm Hg in the placebo group, 159.4 mm Hg in the DEAE-S group, 62.8 mm Hg in the FP group, and 121.1 mm Hg in the FP + DEAE-S group (P =.001, 1 way analysis of variance [ANOVA]). The differences between the control vs DEAE-S groups, placebo vs DEAE-S groups, and FP vs FP + DEAE-S groups were significant (P<.05, t test). Mean hydroxyproline concentrations were 5.2 microg/mg in the control group, 4.9 microg/mg in the placebo group, 5.6 microg/mg in the DEAE-S group, 4.5 microg/mg in the FP group, and 5.4 microg/mg in the FP + DEAE-S group (P =.09, 1-way ANOVA). The difference between the FP and FP + DEAE-S groups was significant (P =.04, t test). CONCLUSIONS: A positively charged particle, DEAE-S, improves healing of colonic anastomoses in healthy rats and in rats with FP. This inexpensive, nontoxic material is easily applied and deserves further evaluation in gastrointestinal tract healing. PMID- 12093339 TI - Recent experience with preoperative fine-needle aspiration biopsy of thyroid nodules in a community hospital. AB - HYPOTHESIS: The application and reliability of fine-needle aspiration (FNA) biopsy in community hospitals may be less efficacious in the clinical assessment of patients with thyroid nodules than in tertiary referral centers. DESIGN: Retrospective review. SETTING: One community teaching hospital. PATIENTS: One hundred eighty-three patients who underwent thyroidectomy after FNA biopsy. INTERVENTIONS: Preoperative FNA biopsy cytopathologic testing and thyroidectomy and postoperative histopathologic testing. MAIN OUTCOME MEASURE: Preoperative cytopathologic reports were compared with postthyroidectomy histopathologic reports. RESULTS: Thyroid cancer was confirmed postoperatively in 70 patients (38%). An FNA biopsy diagnosis of papillary carcinoma (in 29 patients) correlated with a predictive accuracy of 93% (27 patients). Suspicious for papillary carcinoma (n = 14) correlated with malignancy in 8 patients (57%). Indeterminate follicular lesion (n = 60) correlated with malignancy in 18 patients (30%), of whom 16 (89%) had papillary carcinoma (10 patients had follicular variant) and 2 (11%) had follicular carcinoma. Indeterminate Hurthle cell lesion (n = 20) correlated with malignancy in 7 patients (35%). Atypical cell clusters (n = 5) did not correlate with malignancy. Benign FNA biopsy findings (n = 44) in patients who underwent thyroidectomy for other clinical features correlated with malignancy in 8 (18%). Of 11 patients who underwent thyroidectomy for insufficient number of cells after repeated FNA biopsy attempts, 2 (18%) had carcinoma. CONCLUSIONS: The accuracy of an FNA biopsy of thyroid nodules in a community hospital setting is comparable to results from major endocrine referral centers. An indeterminate follicular lesion was the most common FNA biopsy indication for thyroidectomy and correlated with the presence of differentiated thyroid cancers in 18 (30%) of 60 patients. PMID- 12093340 TI - A randomized, controlled trial of Promogran (a collagen/oxidized regenerated cellulose dressing) vs standard treatment in the management of diabetic foot ulcers. AB - HYPOTHESIS: Promogran, a wound dressing consisting of collagen and oxidized regenerated cellulose, is more effective that standard care in treating chronic diabetic plantar ulcers. DESIGN: Randomized, prospective, controlled multicenter trial. SETTING: University teaching hospitals and primary care centers. PATIENTS: A total of 276 patients from 11 centers were enrolled in the study. The mean age of the patients was 58.3 years (range, 23-85 years). All patients had at least 1 diabetic foot ulcer. INTERVENTIONS: Patients were randomized to receive Promogran (n = 138) or moistened gauze (control group; n = 138) and a secondary dressing. Dressings were changed when clinically required. The maximum follow-up for each patient was 12 weeks. MAIN OUTCOME MEASURE: Complete healing of the study ulcer (wound). RESULTS: After 12 weeks of treatment, 51 (37.0%) Promogran-treated patients had complete wound closure compared with 39 (28.3%) control patientss, but this difference was not statistically significant (P =.12). The difference in healing between treatment groups achieved borderline significance in the subgroup of patients with wounds of less than 6 months' duration. In patients with ulcers of less than 6 months' duration, 43 (45%) of 95 Promogran-treated patients healed compared with 29 (33%) of 89 controls (P =.056). In the group with wounds of at least 6 months' duration, similar numbers of patients healed in the control (10/49 [20%]) and the Promogran (8/43 [19%]; P =.83) groups. No differences were seen in the safety measurements between groups. Patients and investigators expressed a strong preference for Promogran compared with moistened gauze. CONCLUSIONS: Promogran was comparable to moistened gauze in promoting wound healing in diabetic foot ulcers. It showed an additional efficacy for ulcers of less than 6 months' duration that was of marginal statistical significance. Furthermore, Promogran had a safety profile that was similar to that of moistened gauze, with greater user satisfaction. Therefore, Promogran may be a useful adjunct in the management of diabetic foot ulceration, especially in ulcers of less than 6 months' duration. PMID- 12093341 TI - The volume-outcome effect for abdominal aortic surgery: differences in case-mix or complications? AB - HYPOTHESIS: Variation in postoperative complications after abdominal aortic surgery contributes to differences in mortality between high- and low-volume hospitals. BACKGROUND: Hospitals with high surgical volume have been shown to have lower operative mortality rates for complex vascular surgery than those with low volumes. Differences in the rates of complications among hospitals may explain this variation in mortality. METHODS: Adult patients who underwent abdominal aortic surgery in Maryland from 1994 to 1996 (N = 2987) were included. The primary dependent variable was in-hospital mortality and the independent variables included hospital surgical volume, patient case-mix variables, and several specific postoperative complications. Two sequential analyses using multiple logistic regression were performed to determine the relative importance of independent variables in predicting mortality. RESULTS: Hospitals with high surgical volume had a lower mortality rate (5.6%) than those with medium (6.8%) and low (8.7%) volumes (P =.03). In the first multivariate analysis, after adjusting for patient case-mix, having surgery at a high-volume hospital remained associated with a 37% reduction in mortality (odds ratio [OR], 0.63; 95% confidence interval [CI], 0.42-0.92; P =.02). Patients at high-volume hospitals had a decreased relative risk (RR) of several complications: pulmonary failure (RR, 0.45; 95% CI, 0.36-0.55), reintubation (RR, 0.53; 95% CI, 0.44-0.64), pneumonia (RR, 0.74; 95% CI, 0.55-0.99), cardiac complications (RR, 0.63; 95% CI, 0.51-0.78), and shock (RR 0.27; 95% CI, 0.10-0.75). In the second multivariate analysis, which included complications, hospital volume was no longer a significant predictor of mortality. However, several postoperative complications remained significant predictors of mortality. CONCLUSIONS: The effect of hospital volume on mortality after abdominal aortic surgery is attributable to differences in postoperative complications and not preoperative differences in case-mix. Efforts to reduce the rates of postoperative complications may reduce mortality rates at low-volume hospitals. PMID- 12093342 TI - Genetic and histological assessment of surgical margins in resected liver metastases from colorectal carcinoma: minimum surgical margins for successful resection. AB - HYPOTHESIS: There have been few reports on the minimum surgical margins (SMs) required for successful liver resection in patients with colorectal metastases. This minimum requirement may be narrower than the previously recommended margin of 10 mm. OBJECTIVES: To identify the minimum margins by assessing the presence of micrometastases around the tumor using genetic and histological techniques, and to investigate whether SMs are associated with patterns of tumor recurrence or patient survival. DESIGN: Prospective and retrospective studies. SETTING: Tertiary referral cancer center. PATIENTS AND METHODS: Fifty-eight patients who underwent 62 liver resections for hepatic metastasis from colorectal cancer between December 1, 1996, and November 30, 2000, were included in the study. Tissue samples taken from the tumor, surrounding liver parenchyma, and Glisson pedicle near the tumor were tested for K-ras and p53 mutations using the mutant allele-specific amplification method. For the retrospective study on patient outcomes, 194 patients who had undergone liver resections between 1980 and 2000 were analyzed according to their SMs. RESULTS: Of the 62 sets of samples from liver metastases, 39 were positive for K-ras and p53 gene mutations or both. Micrometastases in the liver parenchyma surrounding colorectal metastases were present in 2.0% (4/199) of tested samples and were located within 4 mm of the tumor border. Micrometastases via Glisson pedicle were more common (14.3% [3/21]), but these were also confined to a short distance from the tumor edge (/= 16 years. Patients with a LDL cholesterol levels >/= l90 mg/dl or 160-189 mg/dl plus at least two risk factors fulfilled the qualitative criteria for the lipid-lowering drug therapy; patients with a risk >/= 20% in 10 years fulfilled the quantitative criteria. The agreement between both qualitative and quantitative criteria was evaluated by means of the Kappa index. RESULTS: 20% of males and 20.1% of females, and 24% of males and 33.1% of females fulfilled the qualitative and quantitative criteria for lipid-lowering drug therapy, respectively. A low (Kappa = 0.30) and a moderate (Kappa = 0.44) agreement was observed among males and females, respectively, regarding qualitative and quantitative criteria. Increased serum cholesterol was the determinant risk factor for fulfilling qualitative criteria (ORadj) y whereas age, increased blood pressure and increased serum cholesterol were determinants for fulfilling the quantitative criteria. CONCLUSION: The results obtained in this study showed a low agreement between qualitative and quantitative criteria proposed by the report of the Ministry of Health Report to lower cholesterol levels by means of drug therapy among healthy people. Clear and unequivocal criteria to decide what persons are candidates for cholesterol lowering drug therapy should be developed. PMID- 12093395 TI - [Metastatic cancer presentation. Validation of a diagnostic algorithm with 221 consecutive patients]. AB - BACKGROUND: The use of a diagnostic algorithm for metastatic cancer presentation (MCP) might enhance the diagnosis of primary tumors amenable to treatment with considerable savings both in time and diagnostic examinations. MATERIALS AND METHODS: From January 1992 to April 1997, all patients admitted with the diagnosis of MCP were prospectively studied. From each patient, a basic study consisting in a clinical interview, complete physical examination, standard blood testing with tumoral markers and chest X-ray were obtained. Patients with a negative basic study were classified as having a metastatic cancer of unknown origin (MUO); in these patients, a protocolized study (abdominal CT scan and mammography among women) were performed. Patients who after the application of the basic and protocolized studies had no primary tumor detected underwent an exhaustive investigation in order to validate the efficiency of the diagnostic algorithm. RESULTS: Two hundred twenty-one patients were included in the study. The mean age of patients was 63 years (range: 23-82). The main symptom was of bone (30%), neurological (24%), thoracic (16%) and abdominal (16%) origin. The basic study was positive for 138 patients (62.4%), with chest X-ray and physical examination yielding the highest number of diagnoses among these patients. The histology of metastases contributed to the definite diagnosis in 31 patients. Only PSA had a high sensitivity and specificity. Eighty-three patients were classified as MUO. The protocolized study diagnosed the primary tumor in 24 patients (30%), 20 by abdominal CT scan and four by mammography; eight of these patients were deemed to be amenable to treatment. The remaining 59 patients underwent an exhaustive study, and a diagnosis was made in 13; nevertheless, none of them was considered candidate for a specific treatment. Finally, 47 patients (21%) remained undiagnosed. The predominant primary tumors included sites at the lung (42%), prostate (6%) and breast (6%). The most common metastatic locations included bone (42%), central nervous system and liver (24%), and the most common histological types were adenocarcinoma (61%) and undifferentiated carcinoma (15%). CONCLUSIONS: Lung cancer and MUO represented 62% of MCP. The basic study oriented in two thirds of cases, and the physical examination and chest X-ray showed the highest diagnostic yield. The histology of metastases and PSA had a key, diagnostic relevance. A protocolized study based on abdominal CT scan and mammography (females) can identify the remaining treatable tumors. PMID- 12093396 TI - [The special attention to re-admitted patients can be effective. Cost-benefit analysis of a new health care model]. AB - BACKGROUND: Patients with chronic conditions are commonly readmitted to hospital. A study was designed to improve their medical care. The cost/benefit analysis is presented to evaluate its efficiency. PATIENTS AND METHODS: Patients attended at the Internal Medicine Department in a hospital area who were readmitted three or more times in a year were included in the study. In a prospective study, patients were attended by means of a specific consultation with upon request care or through telephone care, with the possibility of elective admission. A comparison was made of costs of the new care program with costs of their usual care, and it was correlated with attending results. RESULTS: Patients were followed for 32 months with a 45% decrease in admissions, 50% in visits to the Emergency Department, and 26% in the hospital stay days. The new program involves monthly savings over 5 million pesetas for the attended population. CONCLUSIONS: The proposed health care program decreases the attending needs in these patients and costs derived from hospital care. An efficient alternative is shown. PMID- 12093397 TI - [Centenarian patients attended at a general hospital]. AB - BACKGROUND: To report the social and medical situation of centenarian patients who required emergency hospital care during the last 8 years. METHOD: Retrospective study of patients aged over 100 years attended at the Emergency Department of a general hospital. The percentages of admissions and mortality rates were then compared with those among patients aged over 65. RESULTS: A total of 51 consultations from 41 patients were recorded, with a mean age of 101.2 years. The number of consultations increased gradually with time (p = 0.008). Ninety-three percent of patients lived with their families, predominantly in the rural setting. The most prevalent conditions included the prostatic syndrome (among males), heart failure and chronic obstruction to the airflow. Only 20% of patients had dementia. Sixty-four percent of consultations required hospital admission, with a mortality rate of 20.5% in this group of patients. The most common discharge diagnoses were heart failure and acute cerebrovascular accident. CONCLUSIONS: A gradual increase in hospital care is likely to be excepted among centenarians, who have an acceptable health status, although with a high risk of mortality during hospital admission. PMID- 12093398 TI - [Treatment of inclusion body myositis]. PMID- 12093399 TI - [Difficult-to-control asthma: a diagnostic and therapeutic challenge]. PMID- 12093400 TI - [Diagnosis and therapy of multi-resistant tuberculosis]. PMID- 12093401 TI - [Drug-induced delayed hypersensitivity syndrome]. PMID- 12093402 TI - [Umbilical tumor and constitutional syndrome]. PMID- 12093403 TI - [Pleural effusion and a rose bush thorn in the sputum specimen]. PMID- 12093404 TI - [Tumor at the limb root in a diabetic patient]. PMID- 12093405 TI - [77-year old woman with constipation and chest hematoma]. PMID- 12093406 TI - [Rapid development of pneumonia]. PMID- 12093407 TI - [Fever and eosinophilia as presenting manifestation of an RS3PE syndrome of likely paraneoplastic origin]. PMID- 12093408 TI - [Clinical course and response to therapy in a case of transverse myelitis and SLE]. PMID- 12093409 TI - [Fatal autoimmune hemolytic anemia in a patient with splenic marginal-zone lymphoma with hairy lymphocytes]. PMID- 12093410 TI - [RS3PE syndrome as presenting form of giant cell arteritis]. PMID- 12093411 TI - [Idiopathic? immunodeficiency of T-CD4 lymphocytes. Case Report]. PMID- 12093416 TI - Economic burden of cardiovascular disease associated with excess body weight in U.S. adults. AB - BACKGROUND: Excess body weight (EBW), which continues to become more prevalent, is a clear contributor to cardiovascular disease (CVD), the leading cause of death and disability among U.S. adults. Information on the economic impact of CVD associated with EBW is lacking, however. OBJECTIVE: To estimate the direct medical costs of CVD associated with EBW. METHODS: We conducted a population based analysis of direct medical costs by linking the 1995 National Health Interview Survey and the 1996 Medical Expenditure Panel Survey. The study subjects are adults (aged > or =25 years, excluding pregnant women) in the non institutionalized, civilian population in 1996. RESULTS: The prevalence of CVD among people in the normal weight (body mass index [BMI] > or =18.5 to <25), overweight (BMI > or =25 to <30), and obese (BMI > or =30) groups was 20%, 28%, and 39%, respectively. There were 12.95 million CVD cases among overweight people, more than 25% of which was associated with overweight. There were 9.3 million CVD cases among obese people, of which more than 45% was associated with obesity. This extra disease burden led to $22.17 billion in direct medical costs in 1996 ($31 billion in 2001 dollars, 17% of the total direct medical cost of treating CVD). CONCLUSIONS: The strong positive association between EBW and CVD, and the significant economic impact of EBW-associated CVD demonstrate the need to prevent EBW among U.S. adults. PMID- 12093417 TI - Estimating the cost of unmet HIV-prevention needs in the United States. AB - BACKGROUND: Although new HIV infection cases have dropped from over 160,000 per year in the mid-1980s to 40,000 per year in the 1990s, HIV incidence has been relatively unchanged for a decade. This number of annual incident infections suggests that substantial, unmet HIV-prevention needs continue to fuel the HIV epidemic in the United States. OBJECTIVES: This study estimates the cost of addressing the unmet HIV-prevention needs in the United States and establishes a performance standard by estimating the number of HIV infections that would have to be prevented in order for these programs to be considered cost saving to society. METHODS: Standard methods of cost and threshold analysis were employed in this study. Interventions needed to address unmet behavioral risks include services to reduce sexual risk of HIV infection, services to provide access to sterile syringes for people who cannot stop injecting drugs, HIV counseling and testing, and intensive preventive services to help HIV-seropositive people avoid transmitting the virus to others. RESULTS: If brief interventions are utilized to address sexual behavior risk, the total program cost (over and above current resource levels) is just over $817 million; and if more expensive multisession, small-group interventions are used, the costs increase to over $1.85 billion. However, even the higher-cost program has a threshold of only 12,000 infections that must be prevented in order for the program to be considered a cost saving to society. CONCLUSIONS: Addressing the remaining unmet HIV-preventive needs in the United States will require a substantial commitment of resources. However, even a greatly expanded HIV-preventive program in the United States could pay for itself through savings in averted medical care costs. PMID- 12093418 TI - Self-report of high cholesterol: determinants of validity in U.S. adults. AB - BACKGROUND: Hypercholesterolemia is a major cardiovascular risk factor, and cholesterol awareness is important in both clinical practice and in public health. We evaluated the validity of self-reported hypercholesterolemia and identified determinants of validity. METHODS: The study design was a cross sectional survey, from 1988 to 1994, of adult participants (N=8236) from the Third National Health and Nutrition Examination Survey for whom self-report of hypercholesterolemia and serum measurement were available. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for self-reported hypercholesterolemia were calculated using total cholesterol > or =5.17 mmol/L (200 mg/dL) and/or taking cholesterol-lowering medication as the criterion standard. RESULTS: Overall test characteristics for self-report were sensitivity, 51%; specificity, 89%; PPV, 87%; and NPV, 55%. Sensitivity of self report was higher among older subjects and non-Hispanic whites, specificity was higher among subjects with >12 years of education, PPV was higher in older subjects, and NPV was higher in younger subjects and in those with >12 years of education. Using higher cholesterol thresholds to define hypercholesterolemia led to higher sensitivity, lower specificity, lower PPV, and higher NPV. Sociodemographic and anthropometric predictors of validity were identified by logistic regression. CONCLUSIONS: Due to low sensitivity, self-reported hypercholesterolemia should be used with caution, both during the patient encounter and for surveillance of trends in hypercholesterolemia in the absence of measured cholesterol levels. Specificity is consistently much higher than sensitivity. The high PPV may be of use in certain clinical situations. Such validation studies should form the foundation for future research based on self report. PMID- 12093419 TI - Usefulness of telephone risk factor surveys in the New Mexico border region. AB - BACKGROUND: We sought to assess prevalence of behavioral risk factors and evaluate the usefulness of survey data gathered by telephone in the New Mexico border region. METHODS: This study compared behavioral risk factor data gathered from two probability sample surveys administered in 1998-1999, one by means of a telephone interview and the second face to face. Prevalence estimates for medical care access, injury, and chronic disease risk factors were compared by survey mode, overall, and stratified by income level. RESULTS: Risk factor prevalence estimates based on telephone interviews resemble estimates obtained from face-to face interviews. Although risk factor prevalence estimated from face-to-face interviews were in general slightly greater than those from telephone interviews, none of these differences reached statistical significance. When data for each respective survey were examined by income level, prevalence estimates for comparable income groups in both surveys were in general agreement and estimates within survey increased with declining income. CONCLUSION: In the New Mexico border region, telephone survey data appear to be reasonably valid and offer opportunities for population-based health research. PMID- 12093420 TI - Healthcare system factors and colorectal cancer screening. AB - BACKGROUND: Developing effective programs to promote colorectal cancer (CRC) screening requires understanding of the effect of healthcare system factors on access to screening and adherence to guidelines. METHODS: This study assessed the role of insurance status, type of plan, the frequency of preventive health visits, and provider recommendation on utilization of CRC screening tests using a cross-sectional, random-digit-dial survey of 1002 Massachusetts residents aged > or =50. RESULTS: A broad definition of CRC screening status included colonoscopy or barium enema (screening or diagnostic) within 10 years, flexible sigmoidoscopy (FSIG) within 5 years, and fecal occult blood testing (FOBT) in the past year as options; 51.7% of subjects aged 50 to 64 and 61.5% of older subjects were current. The uninsured had the lowest current testing rate. Among insured participants, type of insurance had little impact on CRC testing; older subjects enrolled in HMOs had marginally higher rates, although not statistically significant. Increased frequency of preventive health visits and ever receiving a physician's recommendation for FSIG or ever receiving FOBT cards were associated with higher rates of CRC screening among both age groups. CONCLUSIONS: Even when broad criteria are used to define current CRC screening status, a substantial proportion of the age-eligible population remains underscreened. Obtaining regular preventive care and receiving a physician's recommendation for screening appear to be potent facilitators of screening that should be considered in designing promotional efforts. PMID- 12093421 TI - The need for surveillance of delay in age-appropriate immunization. AB - BACKGROUND: Vaccination status is assessed nationally in terms of up-to-date status without regard to the age at which recommended doses were actually received. Our study was conducted in 2000-2001 using the most current National Health Interview Survey (NHIS) public use files available. METHODS: Retrospective analysis to determine up-to-date and age-appropriate vaccination status for children aged 25 to 72 months. Five years of pooled data (1992-1996) were obtained from the NHIS Immunization Supplement for children aged 25 to 72 months with immunization data based on written records. The outcome measures used were months of vaccination delay relative to age-appropriate vaccination standard as well as up-to-date vaccination status for the fourth diphtheria-tetanus-pertussis (DTP 4), Polio3, the first measles-mumps-rubella (MMR1) doses, and the 4:3:1 series. RESULTS: Of the 9223 eligible children, 80% were up-to-date for the 4:3:1 vaccination series, but 48% had experienced delays relative to age-appropriate standards. For the DTP4 dose, 85% were up-to-date, although only 46% had received this dose at the appropriate age. Similarly, 90% of children were up-to-date with their Polio3 dose, with 64% receiving this dose at the appropriate age; 96% were up-to-date for the MMR1, and 58% received this dose at the appropriate age. Age appropriate DTP4 vaccination increased by 17 percentage points from 1992 to 1996, whereas up-to-date DTP4 status increased by only 6% during the same period. CONCLUSIONS: Children with up-to-date vaccination status often experienced considerable delay relative to age-appropriate vaccination standards. Consequently, vaccination status measures based solely on up-to-date status tend to understate the degree of underimmunization in a population. National surveillance of age-appropriate vaccination is necessary to identify subpopulations with the greatest prevalence of vaccination delay and to reveal underlying trends that may not be evident through assessments of up-to-date status. PMID- 12093422 TI - Mass mailings have little effect on utilization of influenza vaccine among Medicare beneficiaries. AB - BACKGROUND: We reviewed published and unpublished studies of mass mailings designed to increase utilization of influenza vaccine among Medicare beneficiaries. METHODS: Search of computerized indexes for published studies and the Medicare Peer Review Organization Health Care Quality Improvement Project database for unpublished studies. Study selection criteria were: use of a controlled trial design and use of mass mailings direct to Medicare beneficiaries to increase receipt of influenza vaccinations. Study selection and data extraction were performed in duplicate by physician reviewers with consensus resolution. RESULTS: Six controlled trials of mass mailings were identified. One study, published in the Morbidity and Mortality Weekly Report (MMWR), reported a modest but statistically significant improvement in influenza vaccination rates among patients who received a letter relative to those who did not. This study was cited as justification for mail interventions performed in five subsequent studies, none of which reported clinically meaningful results. None of these five studies was published. CONCLUSIONS: Mass mailings have at best had clinically trivial effects on increasing influenza vaccination among Medicare beneficiaries. Publication bias has contributed to continued use of a relatively ineffective intervention. PMID- 12093423 TI - Risk factors for tick exposure and suboptimal adherence to preventive recommendations. AB - BACKGROUND: Tickborne diseases are the most common vectorborne illnesses in the United States. Understanding risk factors for tick bites and adherence to preventive measures are important in preventing morbidity associated with tickborne disease. METHODS: A random-digit-dialing telephone survey was administered to 1820 residents of 11 counties in Tennessee. RESULTS: Of respondents, 16.4% reported a tick bite within the previous year. Only 29% consistently used insect repellant in high-risk situations, and only 54% always checked their bodies for ticks after leaving potentially infested areas. Nearly one fourth of dog owners removed ticks from their pets with bare hands. Dog ownership, rural residence, and exposure to a farm were associated with increased risk of tick bites. CONCLUSIONS: We identified several groups that were at increased risk of tick bites and might benefit from targeted prevention messages. PMID- 12093424 TI - Depression as a predictor for coronary heart disease. a review and meta-analysis. AB - OBJECTIVE: To review and quantify the impact of depression on the development of coronary heart disease (CHD) in initially healthy subjects. DATA SOURCES: Cohort studies on depression and CHD were searched in MEDLINE (1966-2000) and PSYCHINFO (1887-2000), bibliographies, expert consultation, and personal reference files. DATA SELECTION: Cohort studies with clinical depression or depressive mood as the exposure, and myocardial infarction or coronary death as the outcome. DATA EXTRACTION: Information on study design, sample size and characteristics, assessment of depression, outcome, number of cases, crude and most-adjusted relative risks, and variables used in multivariate adjustments were abstracted. DATA SYNTHESIS: Eleven studies met the inclusion criteria. The overall relative risk [RR] for the development of CHD in depressed subjects was 1.64 (95% confidence interval [CI]=1.29-2.08, p<0.001). A sensitivity analysis showed that clinical depression (RR=2.69, 95% CI=1.63-4.43, p<0.001) was a stronger predictor than depressive mood (RR=1.49, 95% CI=1.16-1.92, p=0.02). CONCLUSION: It is concluded that depression predicts the development of CHD in initially healthy people. The stronger effect size for clinical depression compared to depressive mood points out that there might be a dose-response relationship between depression and CHD. Implications of the findings for a broader bio-psycho-social framework are discussed. PMID- 12093425 TI - Behavior change intervention research in healthcare settings: a review of recent reports with emphasis on external validity. AB - BACKGROUND: Information to judge both the internal and external validity of health behavior research conducted in healthcare settings is vital to translate research findings to practice. This paper reviews the extent to which this research has reported on elements of internal and external validity, with emphasis on the extent to which research has been conducted in representative settings with representative populations. METHODS: A comprehensive review was conducted of controlled interventions for dietary change, physical activity, or smoking cessation conducted in healthcare settings and published in 12 leading health behavior journals between 1996 and 2000. Using the RE-AIM (reach, effectiveness, adoption, implementation, maintenance) framework, the characteristics and results of these studies were summarized to document the extent to which intervention reach, adoption, implementation, and maintenance were reported and what has been learned about each of these dimensions. RESULTS: A total of 36 studies qualified for review. Participation rates among eligible patients were reported in 69% of studies and were generally quite high; in contrast, only 30% of studies reported on participation rates among either healthcare settings or providers. Implementation data were reported in 77% of the studies and these rates were generally high, with the caveat that intervention was often delivered by paid research staff. Long-term maintenance results were reported very consistently at the individual level, but program continuation was almost never reported at the setting level. CONCLUSIONS: We conclude that a much stronger emphasis needs to be placed on the representativeness of providers and settings that are studied. Examples of how this can be done and recommendations for future research are provided. PMID- 12093426 TI - Improving vaccine coverage in communities and healthcare systems. no magic bullets. PMID- 12093428 TI - Infection, multiple organ failure, and survival in the intensive care unit: influence of glutamine-supplemented parenteral nutrition on acquired infection. AB - OBJECTIVE: We investigated the effect of a glutamine-supplemented parenteral nutrition on intensive-care-acquired infection (ICAI) and its relation to outcome. METHODS: We analyzed new data prospectively collected during a double blind, randomized, and controlled trial in an adult general intensive care unit previously reported (Nutrition 1997;13:295). Eighty-four patients were randomized to receive glutamine-supplemented total parenteral nutrition or an isonitrogenous, isoenergetic control. Sepsis was present on admission in 71% of the patients. Clinical and microbiological data were collected on all new infective episodes and associated treatment decisions. Data were analyzed blind to the randomization and study outcome. RESULTS: There was no significant difference in the number of patients developing new infections or in the number occurring during the first 5 d. There was a non-significant trend to increased numbers of infections in those patients receiving the control feed for at least 5 d. In these patients the glutamine recipients showed significantly fewer catheter related infections: 21 versus 12 (P = 0.026). The difference in overall 6-mo mortality was almost completely described by those patients fed for at least 5 d: 9 of 25 versus 18 of 27 using the control nutrition (P = 0.05). Of the deaths in the intensive care unit due to multiple organ failure, 8 of 8 in the glutamine group and 14 of 16 in the control group sustained one or more ICAI and accounted for 38% versus 74%, respectively, of the ICAIs occurring in those patients. In those patients, despite a similar high incidence of colonization with Candida, those receiving glutamine developed fewer Candida infections and none died, whereas six control patients who developed Candida infections died from multiple organ failure (P = 0.02). Survival was not related to the reduced occurrence of the first acquired infection; however, binary logistic regression analysis of glutamine and the incidence of ICAI after starting total parenteral nutrition to outcome showed that only glutamine was significantly associated with improved 6 mo survival (P = 0.027). CONCLUSIONS: In these severely ill patients, parenteral nutrition containing glutamine may not reduce the overall incidence of ICAI, but it may reduce the risk of dying from acquired infections. The improved survival seen at 6 mo appeared related mostly to reduced mortality in the intensive care unit from multiple organ failure in those patients in whom acquired infections are common. PMID- 12093429 TI - Influence of apolipoprotein E phenotype on metabolism of lipids and apolipoproteins after plant stanol ester ingestion in Japanese subjects. AB - OBJECTIVE: We investigated the effect of apolipoprotein E phenotype on changes in plasma levels of lipids and apoproteins by plant stanol ester (PSE) ingestion in Japanese subjects whose diet is low in fat and cholesterol. METHODS AND RESULTS: The effect of PSE-containing spread was studied in a randomized, placebo controlled trial. One hundred five healthy volunteers were enrolled for this study. Apolipoprotein E phenotyping was done in 96 of 105 subjects. We compared plasma levels at the start and end of the test period (4 wk). The daily ingestion of 2 g of plant stanols from the PSE spread significantly reduced plasma levels of low-density lipoprotein cholesterol by 8.9 +/- 6.6% (mean +/- standard deviation) in the E(3) group and 10.4 +/- 8.0% in the E(4) group. The daily ingestion of 2 g of plant stanols from the PSE spread significantly decreased plasma levels of apoprotein B by 5.4 +/- 7.9% in the E(3) group and 8.9 +/- 7.0% in the E(4) group. No further reductions of low-density lipoprotein cholesterol and apoprotein B were observed with 3 g/d of plant stanols from the PSE spread. CONCLUSION: The ingestion of PSE spread significantly reduced plasma levels of total cholesterol, low-density lipoprotein cholesterol, and apoprotein B. However, the response to PSE ingestion was not influenced by apolipoprotein E phenotype. PMID- 12093430 TI - Effects of oral bovine colostrum supplementation on serum insulin-like growth factor-I levels. AB - OBJECTIVES: We investigated whether supplementation with 60 g/d of bovine colostrum affects blood levels of insulin-like growth factor-I (IGF-I) and IGF binding protein-3 in relation to doping testing. Nine endurance-trained men ingested 60 g/d of bovine colostrum for 4 wk. METHODS: Blood and urine were sampled before starting supplementation. After 4 wk urine and blood samples were taken after an overnight fast and 2 h after ingestion of the last portion to study possible acute effects. RESULTS: Blood IGF-I levels before supplementation were (mean +/- standard deviation) 31 +/- 13 nM/L, and no acute effects were observed after 4 wk of supplementation (33 +/- 9 nM/L). Levels of IGF-binding protein-3 were 136 +/- 11 nM/L before supplementation and 135 +/- 16 nM/L after 4 wk of supplementation. Two hours after ingestion of the last portion, the level of IGF binding protein-3 was 131 +/- 19 nM/L, which was not different from baseline values. Drug testing in a laboratory accredited by the International Olympic Committee did not show any forbidden substance before or after 4 wk of supplementation. CONCLUSIONS: Daily supplementation with 60 g of bovine colostrum for 4 wk does not change blood IGF-I or IGF binding protein-3 levels and does not elicit positive results on drug tests. PMID- 12093431 TI - Energy expenditure measured by doubly labeled water, activity recall, and diet records in the rural elderly. AB - OBJECTIVE: The purpose of this study was to determine whether energy expenditure estimated from physical activity and energy intake were equivalent to total daily energy expenditure in an elderly rural population. METHODS: Twenty-seven elderly male (n = 14) and female (n = 13) subjects (mean age, 74 y) were recruited from a rural Pennsylvania population. Over a 2-wk period, total daily energy expenditure was measured by doubly labeled water (TEE) and estimated from 7-d physical activity recall factors multiplied by weight (PA(WT)), estimated basal metabolic rate (PA(BMR)) and resting energy expenditure from indirect calorimetry (PA(REE)), and energy intake from 3-d self-reported diet records (EI). Analysis of variance was used to determine significant within-subject differences in physical activity, energy intake, and energy expenditure. RESULTS: PA(REE) (men: 13.69 +/- 3.23 MJ, women: 9.51 +/- 2.40 MJ) and PA(BMR) (men: 13.69 +/- 2.99 MJ, women: 10.15 +/- 2.21 MJ) were not significantly different from TEE (men: 12.43 +/- 1.63 MJ, women: 9.44 +/- 0.90 MJ). EI (men: 8.66 +/- 2.34 MJ, women: 7.12 +/- 0.93 MJ) was significantly less than TEE, and PA(WT) (men: 17.03 +/- 4.07 MJ, women: 12.86 +/- 3.41 MJ) was significantly greater than TEE. CONCLUSIONS: Whereas 7-d physical activity recall determined with an age- and gender-specific estimate of resting metabolic rate or measured using indirect calorimetry accurately estimated TEE for this group of rural elderly, self-reported diet records consistently underestimated and physical activity recall determined with weight alone consistently overestimated energy expenditure measured by doubly labeled water. PMID- 12093432 TI - Measurement of body fat by dual-energy X-ray absorptiometry and bioimpedance analysis in men with prostate cancer. AB - OBJECTIVE: The aims of this study were to determine the percentage of body fat (%BF) by dual-energy x-ray absorptiometry (DXA) and bioelectrical impedance analysis (BIA) using a standard adult equation and BIA using a standard geriatric equation in a population of older men with prostate cancer and to compare the results from these different methods. METHODS: We conducted a cross-sectional study in 38 men with locally advanced, node-positive, or recurrent prostate cancer and no history of androgen-deprivation therapy. Body composition was evaluated by DXA with the use of a Hologic 4500A densitometer and BIA. BIA %BF was calculated by using standard equations developed for adult and geriatric populations. RESULTS: %BF by DXA, BIA with the standard adult equation, BIA with the standard geriatric equation, and BIA with the age-appropriate equation were 26.7 +/- 5.3%, 22.5 +/- 5.6%, 38.2 +/- 6.9%, and 35.4 +/- 9.6%, respectively. There were statistically significant differences between %BF by DXA and all BIA estimates. By using the methods described by Bland and Altman (Lancet 1986;1(8476):307), the standard adult equation showed the least bias and variability. CONCLUSIONS: In this group of men with prostate cancer, BIA with the standard adult equation provided a reasonable estimate of %BF compared with DXA, although the differences were statistically significant. BIA with the standard geriatric equation, however, markedly overestimated %BF compared with DXA, even when its use was restricted to elderly men. PMID- 12093433 TI - Validation of a new method for estimating resting energy expenditure of non ambulatory tube-fed patients with severe neurodevelopmental disabilities. AB - OBJECTIVE: We assessed the bias and precision of the Arlington Developmental Center (ADC) equations derived from our previous study and the Harris-Benedict equations for estimating resting energy expenditure in non-ambulatory, tube-fed patients with severe neurodevelopmental disabilities. METHODS: Fifteen non ambulatory patients with neurodevelopmental disabilities referred to the nutrition consult service for evaluation of enteral tube feeding via a permanent ostomy who had a steady-state resting energy expenditure measurement performed by indirect calorimetry were included in the study. The predicted energy expenditure values were compared with the measured resting energy expenditure values and evaluated for bias and precision. RESULTS: Both ADC equations were more precise (95% confidence interval [CI]: 9-22% and 10-18% error, respectively) for the total population than the Harris-Benedict equations (95% CI: 17-40% error). The ADC-2 equation was precise (95% CI: 7-15% error) and unbiased (95% CI: -5 to 139 kcal/d) in contrast to the Harris-Benedict equations (95% CI: 23-54% error; bias, +230 to 365 kcal/d) for patients with cerebral palsy and fixed upper extremity contractures. The Harris-Benedict equations were precise and unbiased (95% CI: 3 14% error; bias, -182 to 39 kcal/d) for patients with cerebral palsy with preservation of upper body movement, whereas the ADC equations were biased toward underprediction and associated with greater error (95% CI: -367 to -73 kcal/d and 7-26% error; 95% CI: -379 to -109 kcal/d and 9-27% error, respectively). CONCLUSIONS: The ADC-2 equation was unbiased and more precise in non-ambulatory adult patients with severe neurodevelopmental disabilities and fixed upper extremity contractures, whereas the Harris-Benedict equations were more precise and unbiased for those with preservation of limited functional and non-functional upper extremity movement. PMID- 12093434 TI - Weight-losing HIV-infected patients on recombinant human growth hormone for 12 wk: a national study. AB - OBJECTIVE: In patients with human immunodeficiency virus, body weight (BW) loss, due mainly to loss of fat-free mass, is associated with progression of disease and mortality. Recombinant human growth hormone (rhGH) may promote BW gain by restoring FFM. METHODS: We investigated the results of adding to highly active antiretroviral therapy of routine rhGH treatment in 34 patients with human immunodeficiency virus who had lost 5% to 20% of their usual BWs. They were recruited by their physicians in Switzerland and were instructed to self administer the drug. Patients were given 6 mg of rhGH each day for 12 wk. BW and body composition, measured by bioelectrical impedance analysis (50 kHz, tetrapolar), were recorded at baseline and at 4, 8, and 12 wk of treatment. RESULTS: At week 12, BW gain averaged 3.0 +/- 0.5 kg (P < 0.001), fat-free mass gain was 4.8 +/- 0.5 kg (P = 0.001), and body fat mass loss was 1.8 +/- 0.4 kg (P = 0.008). BW and fat-free mass increases and FM decrease were evident by week 4 and tended to plateau by week 8. Therapy was well tolerated; one patient developed carpal tunnel syndrome. Five patients abandoned the study for reasons unrelated to the rhGH treatment. CONCLUSION: Our data strongly support the use of rhGH in the treatment of unintentional BW loss associated with human immunodeficiency virus. The low rate of dropouts and the low incidence of side effects make the use of rhGH suitable for primary care management. PMID- 12093435 TI - Night eating in obesity: a descriptive study. AB - OBJECTIVE: We investigated the frequency of night eating (NE) among obese patients, its biological correlates, and its relationships with binge eating disorder (BED). METHODS: The eating behaviors of 166 patients enrolled in a weight-loss program were evaluated by a clinical interview specifically designed to detect BED and NE and by the Three Factor Eating Questionnaire. In all cases body weight and resting energy expenditure were measured. RESULTS: Among all obese subjects, NE was observed in 18 cases (15.7%) and BED in 54 cases (32.5%). NE was more frequent among BED than among non-BED patients; however, in the BED patients the Three Factor Eating Questionnaire Disinhibition and Hunger scores were higher than those in non-BED and NE individuals. Further, between NE and non NE subjects with similar body weights, fully comparable resting energy expenditure was observed, indirectly indicating the lack of difference in overall daily food intake. CONCLUSIONS: Although these behaviors may well overlap, the data of this study uphold the concept that NE and BED have different underlying behavioral constructs. PMID- 12093436 TI - Nutrient composition of banked human milk in Brazil and influence of processing on zinc distribution in milk fractions. AB - OBJECTIVE AND METHODS: We measured the contents of fat, protein, lactose, calcium, phosphorus, zinc, iron, copper, and vitamin A in processed mature milk samples (individual, n = 60, and pooled, n = 10) from a reference human milk bank in Brazil and assessed the effect of pasteurization followed by freezing on the nutrient composition and the pattern of zinc distribution in fractions (fat, whey, and casein) of milk samples (n = 15). RESULTS: Mean nutrient concentrations were within expected ranges in mature milk from healthy women, except fat, which was lower. Interindividual variability of nutrient concentrations was high (coefficient of variation, 21-62%) but reduced overall in pooled samples. Processing of milk samples did not affect the nutrient contents but did cause a significant shift (P < 0.04) in the relative distribution of zinc, with a decrease in the whey fraction and an increase in the fat fraction. CONCLUSIONS: Redistribution and possible alterations in the zinc-binding pattern during processing in human milk banks may reduce zinc bioavailability to the infant. PMID- 12093437 TI - Homocysteine, vitamin B(6), and lipid in cardiovascular disease. AB - OBJECTIVES: Elevated homocysteine (Hcy) is considered an independent risk factor for cardiovascular disease (CVD). An elevated plasma Hcy level may interact with conventional CVD risk factors to further increase vascular disease risk. Therefore, we investigated the plasma levels of Hcy, vitamin B(6) status (pyridoxal phosphate and pyridoxal), and lipid profile in patients with CVD. METHODS: Possible associations between sex, age, body mass index (BMI), and waist to-hip ratio (WHR) to levels of plasma Hcy and plasma Hcy to vitamin B(6) status and lipid profile were examined. RESULTS: Plasma Hcy level, body mass index, and waist-to-hip ratio were significantly higher in patients with CVD than in controls. Male CVD patients had significantly higher plasma Hcy levels than did female patients. Plasma levels of pyridoxal phosphate and total B(6) aldehyde were significantly higher in male than in female patients. Plasma Hcy levels of patients did not correlate to their plasma vitamin B(6) status or to their lipid profiles. Plasma Hcy level correlated positively with age, body mass index, and waist-to-hip ratio (P < 0.0001). CONCLUSIONS: This suggested that patients with CVD have higher levels of plasma Hcy that are influenced by sex, age, body mass index, and waist-to-hip ratio and not by their plasma vitamin B(6) status and lipid profiles. PMID- 12093438 TI - Total parenteral nutrition delays platelet engraftment in patients who undergo autologous hematopoietic stem cell transplantation. AB - OBJECTIVES: One of the major challenges in the post-transplant period is nutrition. In this prospective, non-randomized study, total parenteral nutrition (TPN) was given to 31 patients and partial parenteral nutrition (PPN) was given to 30 patients undergoing autologous hematopoietic stem cell transplantation for solid tumors or hematologic malignancies to compare the effects of these parenteral nutrition modalities on post-transplant hematological engraftment, blood chemistry, and supportive therapy requirements. METHODS: All patients in the TPN group and 17 patients in the PPN group received growth factor in the post transplant period. Both groups did not differ with respect to sex, age, and reinfused CD34(+) cell numbers. RESULTS: After transplantation body mass index and body weight decreased significantly in both groups (P < 0.001). Whereas serum albumin concentrations did not decrease significantly in the TPN group, it fell markedly in the PPN group at the end of parenteral nutrition (P = 0.019). After parenteral nutrition, blood chemistry was also remarkable for serum urea and glucose levels, which were elevated significantly in the TPN group (P < 0.001 and P = 0.03, respectively). Patients receiving TPN had a higher incidence of positive microbial cultures and clinical infection than did patients receiving PPN (64.5% versus 40%, P = 0.05). The most striking result was a delay in platelet engraftment for the TPN group compared with the PPN group (15.54 and 12.93 d, respectively; P = 0.014). This difference was also noted in patients using growth factor in the PPN group (P = 0.017). Parallel to these results, platelet transfusion requirement increased in the TPN group compared with the PPN group (1.93 versus 1.16 U, P = 0.004). Both groups were unremarkable for leukocyte recovery and red blood cell transfusion requirement. CONCLUSIONS: Consequently, TPN has some pitfalls of hyperglycemia, infection tendency, delayed platelet engraftment, and increased platelet transfusion requirement. Therefore, it should not be used as a standard nutrition support for patients undergoing autotransplantation. PMID- 12093439 TI - Relation of cord serum levels of growth hormone, insulin-like growth factors, insulin-like growth factor binding proteins, leptin, and interleukin-6 with birth weight, birth length, and head circumference in term and preterm neonates. AB - OBJECTIVES: Fetal growth process is governed by multiple factors. We investigated the relation of insulin-like growth factors (IGFs), IGF binding proteins (IGFBPs), leptin, and interleukin-6 (IL-6) with intrauterine growth in preterm and term neonates. METHODS: Thirty-eight preterm and 43 term neonates were recruited. Anthropometric measures were recorded and umbilical cord blood samples were collected at birth. RESULTS: Birth weight (BW), birth length (BL), ponderal index, head circumference (HC), and cord serum levels of albumin, prealbumin, retinol-binding protein (RBP), total and free IGF-I, IGF-II, IGFBP-3, acid-labile subunit (ALS), and leptin were significantly lower, whereas levels of IGFBP-1, IGFBP-2, and IL-6 were significantly higher in preterm than in term neonates (P < 0.05). Total and free IGF-I, ALS, and leptin had significantly positive correlations, whereas IGFBP-2 had a significantly negative correlation, with BW and BL in preterm plus term neonates. Forward stepwise multivariate regression analysis showed that gestational age (GA), IGFBP-2, leptin, and free IGF-I are significant predictors of BW; GA, IGFBP-2, ALS, transferrin, and leptin are significant predictors of BL; and GA and free IGF-I are significant predictors of HC in preterm and term neonates. CONCLUSIONS: Our results suggest that IGF-I, IGF II, IGFBP-2, ALS, and leptin play important roles in intrauterine growth. PMID- 12093440 TI - Early enteral supply of fiber and Lactobacilli versus conventional nutrition: a controlled trial in patients with major abdominal surgery. AB - OBJECTIVE: Early enteral nutrition with fiber-containing solutions plus Lactobacillus may reduce bacterial translocation and minimize the incidence of infections after surgery. METHODS: In a prospective, randomized trial in three groups (n = 30/group) of patients after major abdominal surgery, we compared our previous regimen with parenteral nutrition or fiber-free enteral nutrition (group A) with enteral fiber-containing nutrition with living Lactobacillus (group B) and heat-killed Lactobacillus (group C). The main endpoint was the development of bacterial infection. Other analyzed parameters were the durations of antibiotic therapy and hospital stay, non-infectious complications, side effects of the nutrition, and onset of bowel movement. Routine parameters, nutritional parameters, and cellular immune status in the blood were measured preoperatively and on 1, 5, and 10 d postoperatively. RESULTS: The incidence of infections was significantly lower (P = 0.01) in groups B and C with enteral nutrition containing fibers (10% each) than in group A (30%). Patients in group B received antibiotics for a significantly shorter time (P = 0.04) than did the patients in groups A and C. The length of hospital stay and the incidence of non-infectious complications did not differ significantly. Fibers and lactobacilli were well tolerated. There were no general benefits of living Lactobacillus as opposed to heat-killed Lactobacillus in the entire study population, but benefits were observed in the patients with gastric and pancreas resections, although no statistical analysis was done due to their small numbers. CONCLUSIONS: Early enteral nutrition with fiber-containing solutions reduced the rate of postoperative infections in comparison with parenteral nutrition and fiber-free enteral formula. Addition of living Lactobacillus seemed to increase the benefits in patients with gastric and pancreatic resections. PMID- 12093441 TI - Muscle strength as a predictor of loss of functional status in hospitalized patients. AB - OBJECTIVE: The incidence of protein-calorie malnutrition in hospitalized adult patients can reach 30% to 50% and adversely affect clinical outcome. We evaluated the efficacy of muscle strength to predict functional derangement and detect early changes in nutrition status in hospitalized patients. METHODS: Patients hospitalized at medical and surgical wards from two different hospitals in Santiago, Chile, were studied during their hospital stay. Subjective Global Assessment of nutrition status and laboratory parameters were measured on admission. Anthropometric measures, handgrip dynamometry, and maximal inspiratory and expiratory pressures were measured on admission and discharge. The Karnofsky index was used to assess functional status. Twice weekly, caloric balance was calculated with indirect calorimetry and assessment of dietary intake. RESULTS: From the initial selection of patients (n = 70), 50 patients (26 men and 24 women) completed the study. Median hospital stay was 10 d. Subjective Global Assessment was associated with anthropometric data, handgrip dynamometry, and serum levels of total proteins. Patients in whom functional status declined during hospital stay, on admission had lower left handgrip strength, a worse Subjective Global Assessment classification, were older, and had lower fat mass. No association between caloric balance during hospital stay and changes in muscle strength was observed. CONCLUSIONS: Subjective Global Assessment, handgrip strength, and fat mass were good predictors for the decline in functional status during hospital stay. No association between caloric balance and changes in muscle strength was observed. PMID- 12093442 TI - Glycemic and insulinemic responses of nondiabetic healthy adult subjects to an experimental acid-induced viscosity complex incorporated into a glucose beverage. AB - OBJECTIVE: An acid-induced-viscosity (I-V) complex containing alginate, citrate, and insoluble calcium was incorporated into a glucose-based beverage. We hypothesized that the acid I-V beverage would become viscous in the stomach (due to the solubilization of calcium and its interaction with alginate and citrate) and would blunt glycemia. METHODS: Thirty subjects were used in a double-masked, placebo-controlled crossover study evaluating the acid I-V complex. The placebo was a glucose-based beverage that had a similar total dietary fiber level and initial viscosity (Control). After a 12-h overnight fast, serum glucose and insulin were monitored over a 3-h postprandial period. RESULTS: The postprandial mean peak incremental change from baseline in serum glucose tended (P < 0.06) to be lower for the acid I-V product. The net incremental area under the curve (AUC) for serum glucose was reduced 75% (P < 0.01) by the acid I-V product, which was due mainly to an increased undershoot. The mean peak incremental change from baseline in serum insulin was higher (P < 0.05) for the acid I-V product. Net incremental AUC for serum insulin did not differ (P > 0.20) between products. CONCLUSIONS: Results of this study suggested that the acid I-V complex may attenuate the postprandial glycemic response to a glucose challenge in healthy subjects. PMID- 12093443 TI - Fatty acid compositions of serum phospholipids of postmenopausal women: a comparison between Greenland Inuit and Canadians before and after supplementation with fish oil. AB - OBJECTIVES: We compared serum phospholipid fatty acid compositions, in particular the status of omega-3 polyunsaturated fatty acid (PUFA), of postmenopausal Greenland Inuit women and postmenopausal Canadian women at baseline and after supplementing the Canadian women with a fish-oil product. METHODS: Fasting serum samples were collected from 15 Inuit subjects from Greenland and 16 non-Inuit subjects from Canada. In addition, eight Canadian subjects provided fasting serum samples after completing a long-chain omega-3 PUFA intervention (2.4 g of eicosapentaenoic acid [EPA] plus 1.6 g of docosahexaenoic acid [DHA] per day) for 28 d. Fatty acid compositions of serum phospholipids of the samples were determined and compared by one-way analysis of variance. RESULTS: In comparison with the Greenlanders, baseline Canadian women had 73% and 46% less EPA (20:5omega-3) and DHA (22:6omega-3), respectively, and 32% and 91% more linoleic acid (LA; 18:2omega-6) and arachidonic acid (AA; 20:4omega-6), respectively. The omega-3 supplementation in Canadian women increased DHA and decreased LA levels to approach those in Greenland Inuit and raised EPA levels to surpass (45% higher) those in Greenland women (P < 0.0001). In contrast, AA was only moderately lowered (by 16% overall) such that AA levels remained 62% higher in the supplemented Canadians than in the Greenlanders (P < 0.0001). CONCLUSIONS: Short-term EPA plus DHA supplementation of postmenopausal North American women can mimic the high EPA and DHA levels and lower LA levels in corresponding Inuit women but not the markedly lower levels of AA. The present findings also support the hypothesis of genetically decreased Delta5-desaturase potential in the Greenland Inuit compared with Canadian postmenopausal women. PMID- 12093444 TI - Effects of arginine-enriched total parenteral nutrition on inflammatory-related mediator and T-cell population in septic rats. AB - OBJECTIVES: Previous reports have shown that oral arginine (Arg) has immune enhancing properties in injury. However, the effects of parenterally infused Arg on sepsis are not well understood. We used a septic rat model to study Arg infusion in inflammatory-related cytokines and blood T lymphocyte population in vivo. METHODS: Rats with internal jugular catheters were assigned to one of two groups. Both groups received isonitrogenous total parenteral nutrition (TPN) supplemented with 270 mg of nitrogen per kilogram per day as Arg or glycine (Gly). TPN provided 270 kcal/kg of body weight, and the kilocalorie:nitrogen ratio was 143:1. TPN was maintained for 5 d plus 2, 4, or 6 h or 6 d, according to the scheduled deaths of the rats. On day 5, sepsis was induced by cecal ligation and puncture (CLP). After CLP for 2, 4, 6, and 24 h, rats were killed. RESULTS: The results showed that interleukin-1beta and tumor necrosis factor alpha concentrations in peritoneal lavage fluid at 6 h and interleukin-6 levels at 24 h after CLP in the Gly group were significantly higher than those in the Arg group. The T-lymphocyte population in blood showed that CD8(+) suppressor T cell number was significantly higher in the Gly group than in the Arg group at 6 h after CLP. The blood CD4(+):CD8(+) ratio was significantly higher in the Arg group than in the Gly group at 24 h after CLP. A negative nitrogen balance was observed in the Arg and Gly groups after CLP; there was no significant difference in nitrogen balance between the septic groups. No difference in survival rate at 24 h after CLP was observed between the groups. CONCLUSIONS: The results showed that, compared with the Gly group, TPN preinfused with Arg reduces the production of inflammatory mediators at the site of injury and that cellular immunity is enhanced at 24 h after CLP. Parenterally administered Arg had no beneficial effect in preventing nitrogen loss and improving survival in septic rats. Whether Gly has specific effects that reduce the effects of Arg require further investigation. PMID- 12093445 TI - Ingestion of water-soluble soybean fiber improves gastrectomy-induced calcium malabsorption and osteopenia in rats. AB - OBJECTIVE: Total gastrectomy produces calcium malabsorption and osteopenia. We examined the effects of feeding water-soluble soybean fiber (WSSF), a highly fermentable dietary fiber with low viscosity, on calcium absorption and bone characteristics in totally gastrectomized rats. METHODS: Male Sprague-Dawley rats were assigned to two groups: sham operation and gastrectomy. After the postoperative recovery period, rats of each group were fed diets with or without WSSF (50 g/kg diet) for 4 wk. RESULTS: Net calcium absorption and femoral dry weight and calcium content and maximum breaking force were lower in the gastrectomy groups than in the sham groups. In the gastrectomy groups, calcium absorption and these femoral parameters were higher in rats fed the WSSF diet than in rats fed the WSSF-free diet. Feeding WSSF increased the concentrations of short-chain fatty acids and soluble calcium and decreased the pH in cecal contents. Net calcium absorption in gastrectomized rats correlated positively with the concentration of total short-chain fatty acids (the sum of acetic, propionic, and butyric acids) in cecal contents (r = 0.545, P < 0.05) and negatively with cecal pH (r = -0.628, P < 0.05). CONCLUSIONS: These findings indicate that ingestion of WSSF partly prevents the diminished calcium absorption after total gastrectomy in rats, resulting in the improvement of postgastrectomy osteopenia. The increase of calcium absorption may be associated with cecal fermentation of WSSF. PMID- 12093446 TI - Serum leptin concentrations after surgery in young rats. AB - OBJECTIVES: We describe changes in serum leptin concentrations after surgery (laparotomy, partial hepatectomy, splenectomy, and unilateral nephrectomy) in young male rats. Because we presumed that pain and inflammation would influence food intake and subsequent leptin release, laparotomized and partially hepatectomized rats were treated with ibuprofen. METHODS: Preoperative blood samples were taken from the retroorbital sinuses, and postoperative blood samples were taken from the aorta, vena portae, and vena cava 18 h after surgery. Serum leptin concentrations were estimated by radioimmunoassay. RESULTS: Pre- and postoperative serum leptin concentrations did not differ significantly in rats with partial hepatectomy (5.1 +/- 0.4 versus 4.2 +/- 1.1 microg/L), splenectomy (6.1 +/- 0.4 versus 5.0 +/- 0.6 microg/L), or unilateral nephrectomy (4.1 +/- 0.7 versus 5.0 +/- 1.3 microg/L). Significant decreases in serum leptin were observed after laparotomy (8.1 +/- 0.9 versus 3.7 +/- 0.5 microg/L), laparotomy plus ibuprofen treatment (8.9 +/- 1.6 versus 3.1 +/- 0.4 microg/L), and partial hepatectomy plus ibuprofen treatment (5.4 +/- 0.6 versus 3.2 +/- 0.3 microg/L). We observed no significant differences in serum leptin concentrations measured in different regions of the body. CONCLUSIONS: Surgical interventions in young rats are accompanied by decreases in serum leptin. The liver, spleen, and kidney may participate in leptin clearance. Ibuprofen treatment leads to additional decreases in serum leptin concentrations. PMID- 12093447 TI - Dietary fat composition alters pulmonary function in pigs. AB - OBJECTIVES: We investigated the effect of various dietary fats on pulmonary surfactant composition and lung function changes that occur before and after endotoxin infusion in pigs. METHODS: Eighteen pigs were assigned to three groups (n = 6 per group) to receive a diet of protein (20% of calories), carbohydrate (20% of calories), and fat (40% of calories). In one group the fat content consisted entirely of palmitic acid. In the second group, fat came from Intralipid, which provided predominantly linoleic acid. The third group was fed fish oil. Pigs were maintained on these diets for 21 d before the experiment. Cardiovascular and pulmonary functions were determined on day 22. Pigs then were infused with endotoxin (80 mg. kg(-1). min(-1)) until the pulmonary arterial pressure reached a pressure similar to that found in trauma victims (45 to 50 mmHg). Cardiovascular and pulmonary function tests were then repeated, the animals killed, and the lungs removed for study. RESULTS: Compliance was reduced in the linoleate and fish-oil groups compared with the palmitate group before and after endotoxin. Compliance changes in pigs fed the linoleate and fish-oil diets were consistent with significant increases in lung wet:dry weight ratios, increased CO(2) retention, histologic evidence of vascular congestion, intra alveolar edema, and alveolar septa thickening. Changes in surfactant phosphatidylcholine composition between groups were consistent with the notion that increased unsaturated fatty acids could affect surfactant function. CONCLUSIONS: We concluded that the common practice of providing calories in the form of polyunsaturated fatty acids to critically ill patients carries the risk of being detrimental to lung function. PMID- 12093449 TI - Use of amino acids as growth hormone-releasing agents by athletes. AB - Specific amino acids, such as arginine, lysine and ornithine, can stimulate growth hormone (GH) release when infused intravenously or administered orally. Many individuals consume amino acids before strength training workouts, believing this practice accentuates the exercise-induced GH release, thereby promoting greater gains in muscle mass and strength. The GH response to amino acid administration has a high degree of interindividual variability and may be altered by training status, sex, age, and diet. Although parenteral administration consistently leads to increased circulating GH concentration, oral doses that are great enough to induce significant GH release are likely to cause stomach discomfort and diarrhea. During exercise, intensity is a major determinant of GH release. Although one study showed that arginine infusion can heighten the GH response to exercise, no studies found that pre-exercise oral amino acid supplementation augments GH release. Further, no appropriately conducted scientific studies found that oral supplementation with amino acids, which are capable of inducing GH release, before strength training increases muscle mass and strength to a greater extent than strength training alone. The use of specific amino acids to stimulate GH release by athletes is not recommended. PMID- 12093448 TI - Effects of soy protein supplemented with methionine on blood lipids and adiposity of rats. AB - OBJECTIVE: The effects of soy protein isolate (SPI) versus casein on blood lipids and adiposity were investigated in rats fed methionine-equivalent diets. METHODS: Twenty-eight male Sprague-Dawley rats (230 to 250 g) were assigned in equal numbers to groups consuming SPI- or casein-based diets (20%) supplemented with L methionine. After 28 d, blood was collected for triacylglycerol, total cholesterol, and high-density lipoprotein cholesterol assessment and epididymal fat pads were weighed. RESULTS: Food intake (519 +/- 90 versus 490 +/- 115 g), weight gain (144 +/- 35 versus 133 +/- 28 g), food efficiency ratio (0.29 +/- 0.09 versus 0.28 +/- 0.06), epididymal fat pad weights (5.409 +/- 2.076 versus 4.768 +/- 1.867 g), and serum concentrations of triacylglycerol (96.3 +/- 41.8 versus 93.4 +/- 37.4 mg/dL) and high-density lipoprotein cholesterol (32.6 +/- 7.4 versus 33.8 +/- 4.4 mg/dL) were similar between the casein and SPI groups, respectively. However, total cholesterol (73.8 +/- 17.8 versus 59.3 +/- 11.9 mg/dL) concentration was higher for the casein-fed rats than for the SPI-fed rats, respectively (P < 0.05). CONCLUSIONS: These results suggest that methionine supplementation may eliminate the decreased fat deposition previously ascribed to soy protein; however, methionine did not abolish the commonly observed hypocholesterolemic effects of soy. PMID- 12093450 TI - Abrupt and complete occlusion of tumor-feeding vessels by gamma-linolenic acid. PMID- 12093451 TI - Antibacterial activity of citrate and acetate. AB - OBJECTIVE: We evaluated citrate salt, acetate salt, and their combinations for antibacterial activity against a sample of common pathogens. METHODS: Bacterial suspensions were added to serial microdilutions of the salts in broth, with final cell concentrations of 10(4-5) colony-forming units per milliliter. After overnight incubation at 35 degrees C, the minimum inhibitory concentration was recorded. Bactericidal activity was screened by quantitative subcultures from the minimum inhibitory concentration dilution. RESULTS: Citrate salt was active against gram-positive species and Candida albicans but showed little activity against gram-negative species; acetate salt showed the opposite results. Their combination did not show synergism or antagonism. CONCLUSION: It may be feasible to take advantage of the different antibacterial spectra of these two agents and combine them for possible application such as food or medical preservative agents. PMID- 12093452 TI - Is type 2 diabetes mellitus a disorder of the brain? AB - I propose that type 2 diabetes mellitus is due to damage to neurons in the ventromedial hypothalamus or to a defect in the action or properties of insulin or insulin receptors in the brain. These neuronal abnormalities are probably secondary to a marginal deficiency of long-chain polyunsaturated fatty acids during the critical periods of brain growth and development. Hence, supplementation of adequate amounts of long-chain polyunsaturated fatty acids during the third trimester of pregnancy to 2 y postterm can prevent or postpone the development of diabetes mellitus. PMID- 12093454 TI - Serum long-chain polyenoic fatty acids in postmenopausal women. PMID- 12093453 TI - Food and nutrient intakes by pregnant Nigerian adolescents during the third trimester. AB - We assessed the food and nutrient intakes of pregnant adolescent Nigerians during the third trimester of pregnancy. The study was hospital based and the subjects consisted of 101 adolescents (47 urban and 54 rural) and 20 controls (primigravidae, 20 to 25 y old) who attended antenatal clinics. The 3-d weighed intake technique was used to obtain information on food intake. For all cohorts, the results showed that the bulk of the foods derived from three main food groups, namely roots or tubers, cereals, and legumes. Older adolescents (17.5 to 19 y) in urban areas had the highest energy intake (5.9 MJ/d), and the younger urban adolescents (15 to 17 y) had the lowest (5.2 MJ/d) energy intake. Mean protein intake levels for urban adolescents (37.7 g/d) did not differ significantly (P > 0.05) from mean intake levels (35.3 g/d) for rural subjects. Dietary iron intake was similar in all cohorts including controls, and only 32.3% to 39.3% of the recommended daily allowance for iron was met by the subjects. Approximately 30% of subjects individually achieved 100% of the recommended levels of zinc. Mean intakes as percentages of the recommended daily allowance by adolescent subjects were 50.7% for calcium, 29.2% for copper, and 28.5% for folate. There is urgent need to monitor the nutrition status of pregnant women in the area. PMID- 12093455 TI - Pulmonary dysfunction and the integrity of the pulmonary surfactant system. PMID- 12093456 TI - Nutritional management of disabled patients--a new formula for success? PMID- 12093457 TI - Amino acids and growth hormone manipulation. PMID- 12093458 TI - Hypocholesterolemic effect of soy protein. PMID- 12093459 TI - Intraportal nutrition: are there indications for clinical relevance? PMID- 12093461 TI - Reflections on current issues concerning the stability of parenteral nutrition mixtures. PMID- 12093460 TI - A pilot study of the safety and efficacy of supplemental arginine to enhance immune function in persons with HIV/AIDS. AB - OBJECTIVE: We collected preliminary data on the safety and efficacy of supplemental arginine to improve natural killer cell cytotoxicity in a sample of persons with human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS). METHODS: In a randomized, double-blind, placebo-controlled pilot study in an academic medical center-based infectious disease clinic, 11 clinically stable, HIV-infected adults had been treated with highly active, antiretroviral therapy and had HIV plasma RNA levels of less than 10 000 copies/mL. Participants were randomly assigned to receive 19.6 g of arginine/d (n = 6) or placebo (n = 5) for 14 d. Plasma HIV RNA levels, neuropsychologic functioning, and self-reported adverse events were analyzed for safety of treatment. Efficacy was measured by natural killer cell cytotoxicity. RESULTS: None of the participants experienced any adverse clinical, virologic, or neuropsychologic events that necessitated withdrawal from the study. The arginine supplemented group showed a mean natural killer cell cytotoxicity increase of 18.9 lytic units, whereas the placebo group showed an increase of 0.3 lytic units. This difference was not statistically significant (P = 0.79). CONCLUSIONS: Short-term arginine supplementation is safe for persons with HIV/AIDS. Additional studies with larger samples and longer periods are warranted to determine the effects of arginine supplementation on other indices of immune function and on clinical outcomes such as intercurrent illnesses. PMID- 12093462 TI - Glucosamine: con or cure? Part II. PMID- 12093463 TI - Statistical methods for assessing agreement. PMID- 12093466 TI - Overnutrition and nutritional deficiency contribute to metabolic syndrome and atherosclerosis in Asian Indians. PMID- 12093468 TI - Desensitization of insulin secretion. AB - Desensitization of insulin secretion describes a reversible state of decreased secretory responsiveness of the pancreatic beta-cell, induced by a prolonged exposure to a multitude of stimuli. These include the main physiological stimulator, glucose, but also other nutrients like free fatty acids and practically all pharmacological stimulators acting by depolarization and Ca2+ influx into the beta-cell. Desensitization of insulin secretion appears to be an important step in the manifestation of type 2 diabetes and in the secondary failure of oral antidiabetic treatment. In this commentary, the basic concepts and the controversial issues in the field will be outlined. With regard to glucose-induced desensitization, two fundamentally opposing concepts have emerged. The first is that desensitization is the consequence of functional changes in the beta-cell that impair glucose-recognition. The second is that long term increased secretory activity leads to a depletion of releasable insulin, often in spite of increased insulin synthesis. The latter concept is more appropriately termed beta-cell exhaustion. The same dichotomy applies to the desensitization evoked by pharmacological stimuli: again the relative contributions of a decreased insulin content versus alterations in signal transduction are in dispute. The action of tolbutamide on beta-cells may be an example of desensitization caused by a lack of releasable insulin since the signaling mechanisms are nearly unchanged, whereas the action of phentolamine, an imidazoline, induces a strong desensitization without reducing insulin content or secretory granules, apparently by abolishing Ca2+ influx. With pharmacological agents it seems that both, alterations in signal transduction and decreased availability of releasable insulin, can contribute to the desensitized state of the beta-cell, the relative contribution being variable depending upon the exact nature of the secretory stimulus. PMID- 12093469 TI - Toward RNase inhibitors: thermodynamics of 2'-CMP/RNase-A binding in multi-ion buffer. AB - Certain ribonucleases (RNases), such as eosinophil-derived neurotoxin, are associated with pathological conditions (e.g. asthma and inflammatory bowel disease) and can even be overtly cyto(neuro)toxic. It has been proposed that small-molecule inhibitors should have therapeutic utility. We used isothermal titration microcalorimetry to characterize reversible inhibitor cytidine 2' monophosphate (2'-CMP) binding to RNase-A in a multi-ion buffer at 37 degrees as a representative system. The estimated parameters were: K(d)=13.9 microM; DeltaG degrees =-6.90 kcal/mol; DeltaH degrees =-15.7 kcal/mol; and DeltaS degrees = 0.028 kcal/mol-K ('enthalpy-driven' interaction). These data should assist drug design of small-molecule inhibitors of homologous RNase catalytic domains. PMID- 12093470 TI - Thiourea toxicity in mouse C3H/10T1/2 cells expressing human flavin-dependent monooxygenase 3. AB - Human flavin-dependent monooxygenase (FMO) isoforms 1 and 3 were expressed by retroviral gene transfer in mouse C3H/10T1/2 cells. FMO function was determined by the sulfoxidation of p-tolylmethylsulfide (TMS). Enzyme activity ranged from 4 to 30 nmol p-tolylmethylsulfoxide (TMSO)/30 min/mg cell protein for FMO 3 clones; for FMO 1 clones, the range was 1-6 nmol TMSO/30 min/mg. Cytotoxicity in these clones after exposure to thiocarbamate compounds was assessed by clonogenic assay. Thiourea (TU), phenylthiourea (PTU), and alpha-naphthylthiourea (ANTU) were toxic to FMO 3 cells but not to parental and FMO 1 clones; 50% toxicity was attained at 1x10(-4) M TU, 5x10(-6) M PTU, and 1x10(-6) M ANTU. Toxicity was observed after a minimum exposure time of 6 hr. Parental cells were resistant to toxicity for exposure times spanning the entire clonogenic assay period (10 days). Ethylene thiourea (ETU) was not toxic to FMO 3 cells, but preincubation with 1x10(-3) M ETU blocked TU toxicity. Reducing GSH levels by preincubation with 1x10(-5) M buthionine sulfoxime (BSO) increased TU sensitivity in FMO 3 cells from 1x10(-4) to 1x10(-6) M to achieve 50% toxicity. BSO also increased the sensitivity of "low expressor" FMO 3 clones to TU, but did not alter the refractoriness of either parental or FMO 1 expressing cells to TU. N Acetylcysteine afforded modest protection to TU toxicity by shifting 50% cytotoxicity for TU from 5x10(-5) to 1x10(-3) M. TU mutagenicity was assayed by the development of ouabain resistance in parental and FMO 3 C3H/10T1/2 cells. Exposure to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG, direct acting mutagen) and TU was executed with and without prior sensitization with BSO. The mutation frequency for MNNG was 76/1x10(6) surviving cells, whereas no mutants were observed for TU-exposed cultures. The results of this study show that, in isolation, the major human hepatic form of FMO is capable of promoting thiocarbamate toxicity. Consistent with the known reactivity of thiocarbamate intermediates with GSH, treatments that alter GSH levels also altered toxicity in either the protective or sensitizing direction. These cell lines expressing variable levels of FMO 3 and TU sensitivity should prove useful as in vitro systems for dissecting the thiocarbamate toxicity pathway. PMID- 12093472 TI - Inhibition of lipopolysaccharide-stimulated nitric oxide production in RAW 264.7 macrophages by a synthetic carbazole, LCY-2-CHO. AB - In activated macrophages, large amounts of nitric oxide (NO) are generated by inducible nitric oxide synthase (iNOS). This is an important mechanism in macrophage-induced cytotoxicity and inflammation. In the present study, a synthetic carbazole compound, 9-(2-chlorobenzyl)-9H-carbazole-3-carbaldehyde (LCY 2-CHO), was found to have an inhibitory effect on lipopolysaccharide (LPS) stimulated NO generation in RAW 264.7 macrophages (IC50 value of 1.3+/-0.4 microM). LCY-2-CHO did not induce cytotoxicity and had a negligible effect on iNOS activity. To explore the mechanism of inhibition of NO generation by LCY-2 CHO, the expression of the iNOS gene was examined. LCY-2-CHO abolished the LPS induced expression of both iNOS protein and mRNA in a parallel concentration dependent manner with IC50 values similar to those required for inhibition of NO generation. LCY-2-CHO did not enhance the degradation of iNOS mRNA. In cells transiently transfected with an iNOS promoter-chloramphenicol acetyltransferase (CAT) reporter construct, LCY-2-CHO attenuated the LPS-induced iNOS promoter activity. However, LCY-2-CHO had no effect on the degradation of IkappaB-alpha or IkappaB-beta, DNA binding activity, or transcriptional activity of nuclear factor kappaB (NF-kappaB). These results indicate that LCY-2-CHO inhibits NO generation via a decrease in the transcription of iNOS mRNA through a signaling pathway that does not involve NF-kappaB activation. PMID- 12093471 TI - Molecular cloning and pharmacological characterization of bovine calcitonin receptor-like receptor from bovine aortic endothelial cells. AB - A complementary DNA encoding calcitonin receptor-like receptor (CRLR) was isolated from a bovine aortic endothelial cell library. The bovine CRLR has 462 amino acids and 92% homology with the human CRLR. In a reverse transcriptase polymerase chain reaction assay, bovine CRLR was found to be widely distributed, including in the heart and lungs. Stable transfection of bovine CRLR in human embryonic kidney cells (HEK-293) resulted in specific high-affinity [125I] rat adrenomedulin (rADM)-binding (dissociation constant=145+/-15 pM). ADM-stimulated adenylyl cyclase activity with an EC50 value of 5.0+/-1.2 nM. The human ADM receptor antagonist hADM(22-52) inhibited [125I]rADM-binding and ADM-stimulated adenylyl cyclase activity. Interactions between bovine CRLR and individual receptor activity modifying proteins (RAMPs) were also investigated. Transient co transfection of bovine CRLR cDNA with human receptor activity modifying protein 1 (hRAMP1) cDNA in HEK-293 cells resulted in the expression of a CRLR that displayed high-affinity binding to calcitonin gene-related peptide. Co transfection of bovine CRLR with human RAMP2 or RAMP3 cDNAs in HEK-293 cells displayed high-affinity ADM receptors. These observations suggest that in the absence of exogenous RAMPs heterologous expression of bovine CRLR results in an ADM receptor phenotype. PMID- 12093474 TI - Characterization of F-actin depolymerization as a major toxic event induced by pectenotoxin-6 in neuroblastoma cells. AB - Pectenotoxins are a group of marine toxins produced by dinoflagellates and formerly included within the group of diarrhetic shellfish poison or toxins (DSP or DST) because of their physico-chemical properties. However, toxicological data on pectenotoxins are still very scarce and its mechanism of action is largely unknown, but toxicity in laboratory animals has been demonstrated by intraperitoneal injection. In this report, we present results of in vitro toxicological assessment of pectenotoxin-6, a derivative of the parental toxin pectenotoxin-2 first isolated from toxic scallops. Results obtained demonstrate an specific time- and dose-dependent depolymerization of F-actin in neuroblastoma cells exposed to pectenotoxin-6 (half-maximal effect about 700 nM at 24 hr). The change in the state of polymerization of actin was not accompanied by other major effects on specific signal transduction pathways or cell survival rate. Pectenotoxin-6 does not modify cytosolic calcium levels either in a calcium containing or calcium-free medium in human lymphocytes. Only when capacitative calcium influx was first activated, the toxin addition significantly decreased the following calcium influx. In these cells, pectenotoxin-6 only modifies cAMP (adenosine 3',5'-cyclic monophosphate) levels in calcium-free conditions. In addition, no effect on cell attachment or apoptosis induction was observed at micromolar concentrations of pectenotoxin-6. Therefore, we conclude that cytoskeletal disruption is a key mechanism of PTX6-induced toxicity in eukaryotic cells. PMID- 12093473 TI - Protein kinase Calpha-dependent increase in Ca2+-independent phospholipase A2 in membranes and arachidonic acid liberation in zymosan-stimulated macrophage-like P388D1 cells. AB - We previously reported that zymosan-stimulated, protein kinase C (PKC)-dependent arachidonic acid liberation occurs with association of Ca2+-independent phospholipase A2 (iPLA2) with the membranes of macrophage-like P388D1 cells. In the present study, the possible involvement of PKC isoforms (alpha, beta, delta, and epsilon) on the increase in iPLA2 was examined. Stimulation of P388D1 cells with zymosan induced increases in iPLA2 activity and protein in the membranes and liberation of arachidonic acid. In the stimulated cells, PKCalpha, PKCdelta, and PKCepsilon, but not PKCbeta, were increased in the membranes. The zymosan-induced increase in iPLA2 activity was suppressed by pretreatment with 4beta-phorbol 12 myristate 13-acetate for 10 hr, by which PKCalpha and PKCdelta, but not PKCbeta and PKCepsilon, were depleted, and by Go6976, a PKCalpha inhibitor, but not rottlerin, a PKCdelta inhibitor. The zymosan-induced release of arachidonic acid was also reduced by the PKC depletion and Go6976. However, stimulation with 4beta phorbol 12-myristate 13-acetate alone did not increase iPLA2 activity in the membranes. Furthermore, the depletion of intracellular Ca2+ also impaired the zymosan-induced increase in iPLA2 activity in the membranes. However, no increase in iPLA2 activity was observed upon stimulation with Ca2+-mobilizing agents (ionomycin or thapsigargin). Cytochalasin D, an inhibitor of actin polymerization, suppressed the zymosan-induced increases in iPLA2 activity and protein in the membranes and the release of arachidonic acid. These results suggest that zymosan stimulates an increase in iPLA2 in the membranes of P388D1 cells probably through activation of PKCalpha in concert with cytochalasin D sensitive events. PMID- 12093475 TI - JM216-, JM118-, and cisplatin-induced cytotoxicity in relation to platinum-DNA adduct formation, glutathione levels and p53 status in human tumour cell lines with different sensitivities to cisplatin. AB - The aim of this study is to establish anti-tumour potency of the new oral platinum drug JM216 and its metabolite JM118 in relation to the platinum (Pt)-DNA adduct formation, glutathione (GSH)-levels, and p53 status in human cancer cell lines with different sensitivities to cisplatin (CDDP). These parameters were studied in the CDDP sensitive human germ cell cancer cell line Tera and the small cell lung cancer cell line GLC4 and their sublines with in vitro acquired CDDP resistance, Tera-CP and GLC4-CDDP, in a human ovarian cancer cell line transfected with mutant p53 (A2780/mt273) and with an empty vector as control (A2780/cmv), and in the intrinsic CDDP resistant human non-small-cell lung cancer cell line SW1573/S1 and colon carcinoma cell line Caco-2. Cytotoxicity was tested with the microculture tetrazolium (MTT)-assay. Pt-DNA adduct levels were assessed immunocytochemically. Quantitative analysis was performed by double fluorescence video microscopy. Results were correlated with GSH levels and p53 status of the cell lines. This study showed that both JM216 and JM118 can partially circumvent intrinsic and acquired resistance to CDDP. Drug-induced cytotoxicity only correlated negatively with GSH levels for JM216 and CDDP in the tested unselected cell lines. At equimolar basis, JM216 induced lower levels of Pt-DNA adducts in the various cell lines than JM118 and CDDP, whereas the JM118-induced amount and pattern of Pt-DNA adducts was comparable to CDDP. No difference in initial Pt-DNA adducts levels was observed between cell lines sensitive, acquired or intrinsic resistant to CDDP suggesting a Pt-resistance mechanism based on tolerance or increased repair, rather than decreased initial Pt-DNA adduct formation. PMID- 12093476 TI - Effects of 5-iodotubercidin on hepatic fatty acid metabolism mediated by the inhibition of acetyl-CoA carboxylase. AB - Diverse mechanisms of action have been proposed for 5-iodotubercidin, although it is widely used as an adenosine kinase inhibitor that consequently interferes with the metabolism of adenosine and adenine nucleotides. Incubation of rat hepatocytes with iodotubercidin produced important effects on lipid metabolism. (i) Both acetyl-CoA carboxylase and fatty acid synthesis de novo were inhibited in parallel by iodotubercidin, with no change in the activity of fatty acid synthase. The inhibition of both activities showed a comparable dependence on iodotubercidin concentration and was accompanied by a similar decrease (about 60%) in the intracellular malonyl-CoA concentration. (ii) Iodotubercidin stimulated palmitate oxidation, although octanoate oxidation was unaffected. However, this effect can be attributed to the decrease of malonyl-CoA concentration and the concomitant relief of the inhibition of carnitine palmitoyltransferase I, because the activity of this enzyme was found unaltered when determined in cells permeabilized with digitonin. (iii) Iodotubercidin also inhibited cholesterol synthesis de novo. Results, thus, indicate that iodotubercidin increases fatty acid oxidation activity of the liver at the expense of lipogenesis, and we suggest that these effects on fatty acid metabolism are mediated by the inhibition of acetyl-CoA carboxylase, probably due to a more than twice increase in the AMP/ATP ratio and the concomitant stimulation of the AMP-activated protein kinase. PMID- 12093477 TI - Mechanisms of growth inhibition in keratinocytes by mercurio-substituted 4',5' dihydropsoralens. AB - Psoralens, together with ultraviolet light A (PUVA), are used in the treatment of epidermal proliferative disorders. Although these compounds can enter cells and photo cross-link DNA, lipids and proteins, including a specific membrane receptor, are also potential targets for the psoralens. To better elucidate the site of action of the psoralens, we have synthesized a family of 5'-mercurio substituted derivatives of 4',5'-dihydropsoralen. These compounds are identified by their heavy metal content and can be used as a model to deliver thiol reactive psoralen derivatives into keratinocytes. The 5'-mercuriopsoralen derivatives were found to be effective inhibitors of keratinocyte growth without photoactivation. The most active compound, 4,8-dimethyl-5'-iodomercuriomethyl-4',5' dihydropsoralen (IC50=10 microM), was also a potent photosensitizer (IC50=0.3 microM). Depletion of keratinocyte GSH with buthionine sulfoximine markedly increased their sensitivity to this analog, both with and without UVA light. In contrast, N-acetyl-L-cysteine partially protected the cells from growth inhibition, indicating that a sulfhydryl-sensitive site is growth limiting and that this target can be photoactivated. Iodomercurio-4',5'-dihydropsoralen was found to form adducts with GSH and cysteine, which were not active without UVA light. Thus, these adducts may also contribute to the photosensitization reactions of the parent compound. Using plasmid DNA unwinding assays, iodomercurio-4',5'-dihydropsoralen was also found to modify DNA, an activity that increased following UVA light treatment. This suggests that DNA damage may contribute to the actions of these psoralens. Taken together, our data demonstrate that there are multiple sites of action for mercuriopsoralens. These compounds may prove useful for understanding the mechanisms of psoralen-induced growth inhibition in the skin. PMID- 12093478 TI - Differentiation of PC12 cells induced by N8-acetylspermidine and by N8 acetylspermidine deacetylase inhibition. AB - Spermidine is one of the simple polyamines found in cells of virtually all living organisms. It undergoes a metabolic conversion to N8-acetylspermidine catalyzed by an enzyme in cell nuclei and is converted back to spermidine by a deacetylase in the cytoplasm. In this study, two different mechanisms were used to produce an elevation in the level of N8-acetylspermidine in PC12 cells: inhibition of N8 acetylspermidine deacetylase and direct addition of N8-acetylspermidine to the cell culture. The increasing intracellular concentration of N8-acetylspermidine was accompanied by signs of PC12 cell differentiation including increased content of dopamine and morphological changes (neurite outgrowths), suggesting a strong and perhaps causal relationship among these effects. This effect on differentiation appears to be specific for N8-acetylspermidine as the addition of other polyamines including spermidine and N1-acetylspermidine did not elicit these changes. Nerve growth factor (NGF) and dexamethasone, commonly used inducers of differentiation in PC12 cells, produced differentiation without measurable changes in N8-acetylspermidine levels, suggesting that different (or multiple) mechanisms may be involved in these differentiation processes. PMID- 12093479 TI - Evidence for the involvement of L-citrulline but not nitric oxide in the proconvulsant action of the precursor L-arginine on picrotoxin-induced convulsions in rats. AB - To determine the role of the metabolites of L-arginine in its actions on picrotoxin-induced convulsions in rats, the concentrations of nitric oxide (NO) and L-citrulline were measured in the brain 30 and 60 min after the administration of L-arginine (1000 and 2000 mg/kg) or of N-nitro-L-arginine methyl ester (L-NAME, 30 mg/kg), an inhibitor of NO synthase. Animals treated similarly were challenged 30 and 60 min later with picrotoxin (5mg/kg), and the time of onset of myoclonus and clonic convulsions and the frequency of convulsions were determined. These parameters were also determined 30 and 60 min after administering L-arginine in L-NAME-pretreated (30 min) animals. Thirty minutes after the administration of L-arginine, the concentrations of both NO and L-citrulline were raised, the onset of myoclonus and clonic convulsions was delayed, and the frequency of convulsions was decreased, indicating the anticonvulsant property of L-arginine. A 60-min treatment of L-arginine produced a further increase in the concentration of L-citrulline but not that of NO and promoted the frequency of picrotoxin-induced convulsions. Pretreatment with L NAME prevented L-arginine from raising the concentrations of both NO and L citrulline; it also promoted the anticonvulsant actions and prevented the proconvulsant actions of L-arginine. These results lead to the conclusion that NO has no involvement in the time-dependent anti and proconvulsant actions of L arginine on the picrotoxin convulsion model, and that L-citrulline seems to have a role in the proconvulsant action of L-arginine. PMID- 12093480 TI - Comparative expression of two alpha class glutathione S-transferases in human adult and prenatal liver tissues. AB - The ability of the fetus to detoxify transplacental drugs and chemicals can be a critical determinant of teratogenesis and developmental toxicity. Developmentally regulated expression of alpha class glutathione S-transferases (GSTs) is of particular interest, since these isozymes have high activity toward peroxidative byproducts of oxidative injury that are linked to teratogenesis. The present study was initiated to examine the expression and catalytic activities of alpha class GST isozymes in human prenatal liver. Northern analysis demonstrated the presence of hGSTA1 and/or A2 (hGSTA1/2) and hGSTA4 steady-state mRNAs in second trimester prenatal livers. Western blotting of prenatal liver proteins provided corroborating evidence via detection of an hGSTA1/2-reactive protein in both cytosol and mitochondria and of hGSTA4-4-reactive protein in mitochondria alone. Catalytic studies demonstrated that prenatal liver cytosolic GSTs were active toward 1-chloro-2,4-dinitrobenzene (a general GST reference substrate), delta5 androstene-3,17-dione (relatively specific for hGSTA1-1), and 4-hydroxynonenal, a highly mutagenic alpha,beta-unsaturated aldehyde produced during oxidative damage and a substrate for hGSTA4-4. Total GSH-peroxidase and GST-dependent peroxidase activities were 9- and 18-fold higher, respectively, in adult liver than in prenatal liver. Multiple tissue array analyses demonstrated considerable tissue specific and developmental variation in GST mRNA expression. In summary, our results demonstrate the presence of two important alpha class GSTs in second trimester human prenatal tissues, and indicate that mitochondrial targeting of GST may represent an important pathway for removal of cytotoxic products in prenatal liver. Furthermore, the relatively inefficient prenatal reduction of hydroperoxides may underlie an increased susceptibility to maternally transferred pro-oxidant drugs and chemicals. PMID- 12093481 TI - Sexual dimorphic expression of mouse hepatic CYP2B: alterations during development or after hypophysectomy. AB - The constitutive expression of CYP2B mRNA in the livers of mice in the prepubertal stage was sex-independent, with CYP2B9 as the principal isoform. During the maturation stage, CYP2B10 was expressed in both sexes, whereas CYP2B9 was diminished markedly in the males, resulting in a sexually dimorphic expression in adult mice. Hypophysectomy eliminated the sexual dimorphism in the mouse CYP2B subfamily by markedly increasing the expression of both CYP2B9 and CYP2B10 in males to levels similar to those in females. PMID- 12093482 TI - Effects of dexamethasone and protein kinase C inhibitors on the induction of bradykinin B1 mRNA and the bradykinin B1 receptor-mediated contractile response in isolated rat ileum. AB - We detected the expression of inducible bradykinin (BK) B1 receptor mRNA in the rat ileum by the reverse transcriptase-polymerase chain reaction (RT-PCR) method, when the isolated ileum was suspended for at least 1 hr in an aerated Tyrode's solution at 37 degrees. The induction of this mRNA was both time- and temperature dependent, and was followed by a contractile response to des-Arg9-BK at around 3 hr of incubation; this response increased in magnitude with time and was maximal at 6 hr. In contrast, the contraction in response to BK and the expression of B2 receptor mRNA were constant throughout this 6-hr incubation period. The contraction due to des-Arg9-BK was selectively suppressed by B1 receptor antagonists, i.e. des-Arg9[Leu8]-BK and des-Arg10-HOE140, but not by the B2 antagonists D-Arg-[Hyp3,Thi5,8,D-Phe7]-BK and HOE140. The inducible des-Arg9-BK contractile response was suppressed by continuous in vitro exposure of the ileum to cycloheximide or actinomycin D, but neither inhibitor affected the contraction induced by BK, suggesting that the B1 receptor could be induced de novo. In vitro and ex vivo treatment of the ileum with dexamethasone suppressed the induction of the contractile response to des-Arg9-BK, but had no significant effect on the expression of B1 receptor mRNA. Some protein kinase C inhibitors, i.e. H7 and calphostin C, suppressed the expression of B1 receptor mRNA and diminished the contractile response to des-Arg9-BK. These results suggest that the de novo synthesis of the B1 receptor in the ileum preparation can be up-regulated at the transcriptional level (a process in which a specific isoform of protein kinase C may be involved). Additionally, these data suggest that the contractile response to des-Arg9-BK involves a process sensitive to some post-transcriptional action of dexamethasone. PMID- 12093483 TI - 3-isobutyl-1-methylxanthine (IBMX) sensitizes cardiac myocytes to anoxia. AB - Cardiac myocytes incubated with 3-isobutyl-1-methylxanthine (IBMX), a nonspecific cyclic nucleotide phosphodiesterase inhibitor, formed rigor complexes under anoxic conditions more readily than cells incubated with other phosphodiesterase inhibitors. Cardiac myocytes were incubated for 1 hr with either (a) no additions, (b) 150 microM zaprinast, or (c) 1 mM IBMX, and then were rendered anoxic for periods up to 60 min. Cells were >80% viable throughout the anoxic period; viability was unaffected by either drug. Rod count decreased more rapidly after the onset of anoxia in the IBMX-treated cells than in control or zaprinast treated cells (11% rods vs. roughly 47% rods after 30 min of anoxia). IBMX treated cell groups also formed more "contracted" myocytes (box-like rods) than their untreated or zaprinast-treated counterparts (50% contracted vs. roughly 27% contracted after 30 min of anoxia). While nucleotide degradation patterns were similar in all experimental groups, the ratio of ATP to ADP was lower in IBMX treated cells than in control or zaprinast-treated cells. The L-type calcium channel was apparently not involved in this phenomenon; while cyclic AMP was elevated in the IBMX-incubated cells, verapamil did not protect IBMX-incubated cells from premature damage by anoxia. Incubation with 8-cyclopentyl-1,3 dipropylxanthine (CDPX), an A1 receptor antagonist, at concentrations up to 1 microM in place of 1mM IBMX did not reproduce the IBMX effect. We concluded that IBMX sensitizes cardiac myocytes to anoxia through a mechanism related to its effect on ATP/ADP, and unrelated to an elevation of intracellular calcium or preconditioning phenomena. PMID- 12093485 TI - Lipid peroxidation induced by indomethacin with horseradish peroxidase and hydrogen peroxide: involvement of indomethacin radicals. AB - Some of the side-effects of using indomethacin (IM) involve damage to the gastric mucosa and liver mitochondria. On the other hand, neutrophils infiltrate inflammatory sites to damage the tissues through the generation of reactive oxygen species by myeloperoxidase. The stomach and intestine have large amounts of peroxidase. These findings suggest that peroxidases are involved in tissue damage induced by IM. To clarify the basis for the tissue damage induced by IM in the presence of horseradish peroxidase (HRP) and H2O2 (HRP-H2O2), lipid peroxidation was investigated. When IM was incubated with liver microsomes in the presence of HRP-H2O2 and ADP-Fe3+, lipid peroxidation was time-dependent. Catalase and desferrioxamine almost completely inhibited lipid peroxidation, indicating that H2O2 and iron are necessary for lipid peroxidation. Of interest, superoxide dismutase strongly inhibited lipid peroxidation, and it also inhibited the formation of bathophenanthroline-Fe2+, indicating that reduction of the ferric ion was due to superoxide (O2-). ESR signals of IM radicals were detected during the interaction of IM with HRP-H2O2. However, the IM radical by itself did not reduce the ferric ion. These results suggest that O2- may be generated during the interaction of IM radicals with H2O2. Ferryl species, which are formed during the reduction of iron by O2-, probably are involved in lipid peroxidation. PMID- 12093484 TI - The direct stimulation of Gi proteins by neuropeptide Y (NPY) in the rat left ventricle. AB - Neuropeptide Y (NPY) is a neuropeptide with high distribution in the cardiovascular system of mammals, where it modulates heart and vessel contractility. In the rat heart, the presence of at least three different NPY receptor subtypes has been hypothesised. Notwithstanding this, receptor activation might not be the only mechanism responsible for the complex cardiac effects of the peptide. In this study, we investigated the effect of NPY on the GTPase activity of G-proteins in the rat left ventricle as a possible alternative mechanism of action for the peptide in the rat heart. Our results show that NPY, but also the neuropeptide fragment (18-36) (NPY (18-36)), stimulated the basal, spontaneous GTPase activity of ventricle membranes only when it was measured under the condition of an absence of Mg2+. This stimulation was resistant to BIBP3226 a non-peptidergic antagonist at Y1 receptors, but it was significantly reduced in membranes treated with selective antibodies against the Gialpha subunits. NPYs effect was concentration-dependent with a maximum of activity at 10nM. At this concentration, NPY (and NPY 18-36) was able to inhibit forskolin (FSK)-induced cyclic adenosine-5'-monophosphate (cAMP) elevation in rat left ventricle slices. Our results assess that NPY in the rat heart is able to activate the GTPase activity of Gi proteins, in a receptor-independent way. PMID- 12093486 TI - Electrochemical evaluation of rhodium dimer-DNA interactions. AB - The interaction of rhodium dimers, including the carboxylates (acetate, propionate, butyrate, trifluoroacetate, citrate and gluconate), amidates (acetamidate and trifluoroacetamidate) and carboxamidate (Doyle catalyst S) with DNA was investigated by electrochemical methods. Differential pulse voltammetry measurements showed, in agreement to literature data, that most of rhodium carboxylates have a higher affinity for adenine than guanine residues. Some differences of reactivity may be correlated with the compound structures and these were helpful in understanding the influence of equatorial ligands on axial coordination mechanisms. The preliminary results might be extended for further studies on quantitative structure activity relationship approaches, highlighting electrochemical methods as a tool for this purpose. PMID- 12093487 TI - Potentiometric determination of polyether ionophores growth promotors in animal feed preparations. AB - The construction and electrochemical response characteristics of four polyvinylchloride (PVC) membrane sensors for the determination of monensin (MN) and salinomycin (SL) were described. The membranes were prepared using 1 wt.% drug, 44 wt.% nitrophenyl octyl ether, 53 wt.% PVC and 2 wt.% lipophilic additive which is the anionic potassium tetra (4-chlorophenyl) borate in sensors 1 and 2 and the cationic nicklo-phenanthroline in sensors 3 and 4. Sensor 1 and sensor 2 show linear responses over concentration range of 10(-3)-10(-5) M drug with cationic slopes of 52.3 and 54.1 mV per concentration decade, respectively. On the other hand sensor 3 and sensor 4 show linear responses over concentration range 10(-4)-10(-5) M drug with anionic slopes of 28.1 and 29.7 mV per concentration decade, respectively. The 4 sensors were successfully applied to the determination of MN and SL in their pharmaceutical products (Premix) with average recoveries of 98.4-100.4+/-2.41-1.97% for sensors 1 and 2 and 97.6-98.8+/ 3.16-3.07% for sensors 3 and 4. The obtained results were compared reasonably well with the data obtained using the USP method (2000). The proposed sensors were also applied for the direct determination of both drugs in animal feed preparations without prior treatment in low levels; 10-1000 microg per 5 g feed. The sensors were successfully used to follow up the drugs concentration in the presence of other growth promotors, other antibiotics and the coexisting fatty acids. PMID- 12093488 TI - Determination of pioglitazone hydrochloride in bulk and pharmaceutical formulations by HPLC and MEKC methods. AB - High Performance Liquid Chromatographic (HPLC) and Micellar Electrokinetic Chromatographic (MEKC) methods have been developed for the determination of pioglitazone, a new englycemic antidiabetic agent. Pioglitazone and its unsaturated impurity were separated by MEKC in less than 7 min using a 43 cm x 50 microm i.d. uncoated fused-silica capillary with extended light path for better sensitivity (25 kV at 30 degrees C) and a background electrolyte (BGE) consisting of 20% acetonitrile (v/v) in 20 mM sodium borate buffer pH 9.3 containing 50 mM sodium dodecyl sulphate (SDS). The influence of various parameters on the separation such as pH of the buffer, SDS concentration, buffer concentration, organic modifiers, temperature and voltage were investigated. The MEKC method was compared with HPLC method using a 5 microm symmetry C18 column (250 x 4.6 mm i.d.) eluted with a mobile phase consisting of a mixture of 50% (v/v) acetonitrile and 10 mM potassium dihydrogen phosphate buffer, adjusting the pH to 6.0 with 0.1 M KOH. The HPLC method is capable of detecting all process related compounds, which may be present at trace levels in finished products. Both methods were fully validated and a comparison was made. The results confirm that the methods are highly suitable for its intended purpose. PMID- 12093489 TI - LC determination of a sulphur mustard decontaminant CC-2 in rat serum. AB - A high-performance liquid chromatographic (HPLC) method was developed for the analysis of N,N'-Dichlorobis (2,4,6-trichlorophenyl) urea (CC-2), a potent sulphur mustard decontaminant, in rat serum. The HPLC analysis, applicable to 0.5 ml volumes of serum, involved double extraction of serum samples with diethyl ether at alkaline pH followed by separation on a RP-18 column and the use of UV detector at 230 nm. The method was sensitive with a limit of quantitation of 10 ng ml(-1) in rat serum and the recovery was always >90%. Excellent linear relationships (r>0.99) were obtained between the measured and added concentration ratios of the serum concentrations over a range of 10-200 ng ml(-1). The precision and accuracy were acceptable as indicated by relative standard deviation ranging from 2.47 to 17.49%, bias values ranging from -4.35 to 13.21%. Moreover, CC-2 was found stable in rat serum even after 3 months of storage at 60 degrees C and being subjected to three freeze-thaw cycles. The assay was found to be sensitive, specific, accurate, precise, and reliable for use in pharmacokinetic studies. PMID- 12093490 TI - Electrochemical behaviour of droxicam: kinetic study in aqueous-organic media. AB - A kinetic study of the hydrolytic decomposition of droxicam was carried out in order to establish the possible pharmacological action of the drug in the organism of the human being. The electrochemical technique used was scan cyclic voltammetry on a HMDE, giving a well-defined peak. Due to the insolubility of droxicam in water, the working media was a methanol-H(2)O (4:96, V/V) mixture. The decomposition product of droxicam is piroxicam, which is also reduced on a mercury electrode giving a well-defined cyclic voltammetric peak. It is proposed that the decomposition of droxicam is controlled by a kinetic equation: v=(1.93 x 10(-3)+78.9 [D]) [D]=k(D)[D]. PMID- 12093491 TI - Quantification of pilsicainide in serum by capillary electrophoresis. AB - A new method for determining pilsicainide concentration in serum by rapid and selective capillary electrophoresis has been developed and validated. For pretreatment, serum was made alkaline and then extracted with diethyl ether. Procainamide was used as an internal standard. Sodium dihydrogenphosphate buffer (pH 2.29; 0.1 M) was used as a running buffer. A fused-silica capillary tube was loaded with a voltage of 25 kV and detection was performed at UV 200 nm. Good linearity (0-2.5 microg/ml) was obtained with the minimum limit of detection being 0.04 microg/ml serum (signal-to-noise ratio, 3:1). The R.S.D. of within-run reproducibility was 0.798-2.32%, that of between-run reproducibility was 4.74 5.12% and the recovery rate was 61-63%. Disopyramide, another anti-arrhythmic drug, was close to pilsicainide in terms of migration time. This method was applied to determination of pilsicainide in serum samples. PMID- 12093492 TI - LC determination of YM466, a new factor Xa inhibitor, in rat and dog plasma. AB - A specific method for the determination of YM466, a novel Factor Xa inhibitor, in rat and dog plasma was developed and validated. YM466 was extracted from plasma by solid-phase extraction and analyzed by UV-HPLC at an absorbance wavelength of 240 nm. The intra-day precision and accuracy ranged from 0.8 to 2.4% and 0.1 to 5.0% in rats, and from 1.6 to 2.4% and 0.0 to 4.1% in dogs, respectively. The lower limit of quantification was 10 ng/ml when 1 ml of plasma was used. No endogenous interference was observed in the plasma of rats and dogs. PMID- 12093493 TI - Physico-chemical studies on the charge-transfer complex formed between sulfadoxine and pyrimethamine with chloranilic acid. AB - Thermodynamic studies on the charge-transfer complexes of sulfadoxine and pyrimethamine with chloranilic acid in non-aqueous 1,4-dioxan were investigated by spectrophotometric method. The absorption maxima for sulfadoxine and pyrimethamine were observed at 500 and 520 nm, respectively. We observed a bathochromic shift in the transitions of these complexes with respect to chloranilic acid, which absorb at 420 nm. The enthalpy of complexation was found to be -3.044 kJ/mole for pyrimethamine and -5.934 kJ/mole for sulfadoxine. Equilibrium constant values were generally high, that is 261.202 l/mole and 344.29 l/mole for pyrimethamine and sulfadoxine, respectively, at 303 K. Free energy values were negative, of the order of -14.420 kJ/mole for pyrimethamine and -15.384 kJ/mole for sulfadoxine, indicating that complex formations were exothermic. These data suggest that the complexes are very stable. Arguments are presented to support the application of charge-transfer interactions in the quantitative analysis of these drugs. PMID- 12093494 TI - Determination and degradation study of haloperidol by high performance liquid chromatography. AB - A specific, high performance liquid chromatographic method was developed for the assay of haloperidol, together with an adequate separation of its degradation products. The method is based on the use of an octadecylsilane stationary phase column under isocratic conditions. The mobile phase consisted of 50 mM sodium phosphate monobasic pH 2.5-acetonitrile-THF-TEA (63:34:3:0.1, v/v/v/v) adjusted with o-phosphoric acid to a pH of 2.5. The degradation was performed in hydrochloric acid, sodium hydroxide and hydrogen peroxide. The main degradation products were identified. Application of the assay of haloperidol in tablet formulations is presented. PMID- 12093495 TI - Screening and optimisation of an ELISA method for the quantitative detection of enterovirus specific RT-PCR products by means of a two-level experimental design. AB - In a previous paper, optimal reaction conditions were determined for the RT-PCR part of a quantitative enterovirus specific RT-PCR ELISA method (J. Pharm. Biomed. Anal., 25 (2001) 131-142). In order to obtain a detection limit as low as possible, the ELISA part of the procedure was optimised as well. This was done by investigating the influence of seven factors at three levels in a multivariate approach. A reflected two-level screening design, derived from a Plackett-Burman design, was used. Optimal reaction conditions were established by calculation and by evaluation of the effects of the factors on the measured absorbance of the ELISA detection. Under these conditions, the linear range and detection limit of the test were determined and compared with the ELISA conditions before optimisation. The optimised RT-PCR ELISA will be used to study a possible longitudinal relationship between enteroviruses and the development of multiple sclerosis and juvenile diabetes. PMID- 12093496 TI - Comparative study of the micellar enhanced spectrophotometric determination of beta-lactamic antibiotics by batch and flow injection analysis using a multisimplex design. AB - A study has been made on the spectrophotometric determination of the beta lactamic antibiotics, amoxicillin and ampicillin, in micellar media using Cu ions as catalyst. Batch and flow injection approaches were compared. Multisimplex design was used to determine the optimal values of the flow injection analysis (FIA) system. Chemical (buffer, pH and Cu(II) concentrations) and physical (flow rate, temperature and reaction coil length) variables were considered. The analytical performance characteristics were as follows: the detection limits for batch and flow-through systems were 2.5 x 10(-7) and 2 x 10(-6) M, respectively, and a relative standard deviation less than 1% was found for both methods. The proposed FIA methodology was satisfactorily applied to the determination of the antibiotics in pharmaceutical formulations. PMID- 12093498 TI - GC/MS evaluation of thyme (Thymus vulgaris L.) oil composition and variations during the vegetative cycle. AB - Capillary GC/MS analysis based on polar and non-polar columns has been applied to evaluation of the volatile oils hydrodistilled from thyme (Thymus vulgaris L.) plants. The adopted methodology has been used to monitor seasonal variations in the composition of the oil obtained from thyme herbs harvested at different periods during the plant vegetative and life cycles. Oils from thyme plants of young (2 years) and old (5 years) cultivations have been evaluated from four and two collections, respectively, effected throughout May/December growth period. Generally, the oil was found to be rich in the active monoterpene phenols (thymol and carvacrol) and their corresponding monoterpene hydrocarbon (HC) precursors (p cymene and gamma-terpinene), which collectively showed synchronized patterns of variation during the different collection periods and in different seasons. The oil from old plant collected in May/June period (0.15% v/w) was characterized by significantly lower levels of monoterpene HCs (mainly gamma-terpinene) and the highest levels of the oxygenated monoterpenes (linalool and borneol), monoterpene phenols (mainly thymol) and their derivatives (mainly carvacrol methyl ether), sesquiterpenes (mainly beta-caryophyllene) and their oxygenated derivatives (e.g. caryophyllene oxide) in comparison with all other samples. A characteristic presence of camphor and thymodihydroquinone was also observed in the old plant oils. On the other hand, the young plant, collected in June/July just before the end of the vegetative cycle, provided the best oil yield (1.2%) with also the highest % content of the monoterpene phenols (thymol: 51.2% and carvacrol: 4%). This latter growth period can represent the best harvest time of young thyme plants in order to obtain an essential oil with better quality and quantity. PMID- 12093497 TI - Simultaneous determination of fexofenadine and its related compounds by HPLC. AB - A simple reversed phase liquid chromatographic (RPLC) method has been developed and subsequently validated for the determination of fexofenadine hydrochloride and its related compounds A and B. The method utilizes a C8 column for the separation and determination of meta-isomer (related compound B). The separation was achieved using an Eclipse XDB C8, 5 microm, 4.6 x 150 mm column and a mobile phase comprising 1% triethylamine phosphate (pH 3.7), acetonitrile and methanol in the ratio 60:20:20 (v/v/v). 5-Methyl 2-nitrophenol has been used as internal standard for the purpose of quantitation of fexofenadine. The described method was linear over a range of 0.7-18.7 microg/ml for related compounds A and B and 60-750 microg/ml for assay of fexofenadine. The relative standard deviation (n=3) was 0.5% for the drug and 3.4% for related compounds. The intermediate precision was 0.79% (n=9) for assay and 5.16% (n=9) for related impurities. The mean recovery of both the related compounds were in the range of 94-103%. Limits of detection (LOD) and quantification (LOQ) for the related compounds A and B were 0.18, 0.12 and 0.56, 0.48 microg/ml, respectively. The precision of the method was checked by F-test using a reported method as reference and the calculated value (1.35) was found to be less than the table value at 95% confidence levels. The obtained results confirm that the method is highly suitable for its intended purpose. PMID- 12093499 TI - Validation of a HPLC quantification of acetaminophen, phenylephrine and chlorpheniramine in pharmaceutical formulations: capsules and sachets. AB - Acetaminophen, phenylephrine and chlorpheniramine are frequently associated in pharmaceutical formulations against the common cold. Their quantification presents several problems. A HPLC method for the simultaneous determination of these compounds in pharmaceutical formulations such as capsules and sachets, including the separation of impurities and excipients has been developed and validated. The selectivity of the method was also tested to be used if phenylpropanolamine hydrochloride were employed instead of phenylephrine. Final chromatographic conditions were a gradient elution, being solvent A: phosphate buffer 40 mM at pH 6.0 and solvent B: acetonitrile. At t=0, the mobile phase consisted of 92% A and 8% B and it changed with a linear gradient during 8 min to 75% A and 25% B. At min 8, it changed to 30% A and 70% B for 5 min and at t=15 min, it returns to the initial conditions (92% A and 8% B) during 1 min remaining at this composition until t=20 min. UV detection was performed at 215 nm for phenylephrine and chlorpheniramine, because at this wavelength sensitivity was higher than in other more characteristic wavelengths and it was necessary for the detection of minor compounds. For acetaminophen 280 nm was employed. Validation parameters permit to consider the method adequate. PMID- 12093500 TI - Determination of lovastatin and simvastatin in pharmaceutical dosage forms by MEKC. AB - A micellar electrokinetic chromatographic (MEKC) method was developed for the quantification of lovastatin and simvastatin, cholesterol lowering agents in pharmaceutical dosage forms. Lovastatin and simvastatin were separated using an electrolyte system consisting of 12% acetonitrile (v/v) in 25 mM sodium borate buffer pH 9.3 containing 25 mM sodium dodecyl sulphate (SDS) with an extended light path capillary (48.5 cm x 50 microm i.d, 40 cm to detector). The method has been validated and proven to be rugged. Calibration curves were linear over the studied ranges with correlation coefficients greater than 0.996. A limit of detection of 3.2 microg/ml and a limit of quantitation of 10.6 microg/ml were estimated for both the drugs. The proposed method was found to be suitable and accurate for the determination of these drugs in commercial formulations. PMID- 12093501 TI - Determination of nicotinamide and 4-aminobenzoic acid in pharmaceutical preparation by LC. AB - A rapid, precise and time saving high performance liquid chromatographic method has been developed and validated for the simultaneous determination of nicotinamide and 4-aminobenzoic acid in pharmaceutical preparation. The method involves isocratic elution of mobile phase through column in a reverse phase chromatography with UV detection at 254 nm. The ranges of quantification for nicotinamide and 4-aminobenzoic acid were 11-34 microg ml(-1) and 37-113 microg ml(-1), respectively. Investigating specificity, linearity, precision, accuracy, robustness and ruggedness performed the validation of the method. PMID- 12093502 TI - Spectrophotometric determination of piroxicam and tenoxicam in pharmaceutical formulations using alizarin. AB - New spectrophotometric procedures have been established for the quantitation of piroxicam and tenoxicam. The procedures are based on the reaction between the examined drug and alizarin (I), alizarin red S (II), alizarin yellow G (III) or quinalizarin (IV) producing ion-pair complexes which can be measured at the optimum wavelength. The optimization of the reaction conditions is investigated. Beer's law is obeyed in the concentration ranges 0.05-2.40 microg ml(-1), whereas optimum concentration as adopted from Ringbom plots was 0.12-2.25 microg ml(-1). The molar absorptivity, Sandell sensitivity, detection and quantification limits are also calculated. The correlation coefficient was >/=0.9990 (n=10) with a relative standard deviation (R.S.D.) of 98.04% for both methods over the linear range. The UV and HPLC methods have been successfully used to determine the I, II and III content in tablets of different origin. PMID- 12093514 TI - Determination of cidofovir in both skin layers and percutaneous penetration samples by HPLC. AB - The aim of this study was to develop a direct, simple and rapid high performance liquid chromatographic method for the determination of cidofovir in both skin layers and percutaneous penetration experiments. Samples were chromatographed on a reversed phase encapped column 250 x 4 mm C(8) LiChrospher Select B. The phase mobile consisted on 3% of acetonitrile and 97% of 1.5 mM of tetrabutylammonium dihydrogen phosphate (TADP) and 3.5 mM of disodium hydrogenphosphate adjusted to pH 6. Detection was at 274 nm and the run time was 14 min. The limit of detection was 0.06 microg/ml. The detector response was found to be linear in the concentration range 0.1-10 microg/ml. This assay is a selective, sensitive and reproducible method for the quantification of cidofovir in skin layers and in the receptor compartment of Franz-type diffusion cells after percutaneous studies. PMID- 12093516 TI - Kinetic spectrophotometric determination of tramadol hydrochloride in pharmaceutical formulation. AB - Two simple and sensitive kinetic methods for the determination of tramadol hydrochloride are described. The first method is based upon a kinetic investigation of the oxidation reaction of the drug with alkaline potassium permanganate at room temperature for a fixed time at 20 min. The absorbance of the colored manganate ions was measured at 610 nm. The second method is based on the reaction of tramadol hydrochloride with 4-chloro-7-nitrobenzofurazan (NBD-Cl) in presence of 0.1 M sodium bicarbonate. The spectrophotometric measurements were recorded by measuring the absorbance at 467 nm, at fixed time at 25 min on thermostated water bath at 90+/-1 degrees C. All variables affecting the development of the colour have been investigated and the conditions were optimised. The absorbance concentration plots in both methods were rectilinear over the range 5-25 and 50-250 microg ml(-1), for the first and second methods, respectively. The two methods have been applied successfully to commercial capsule and ampoule dosage form. The results obtained are compared statistically with those given by the reference spectrophotometric method. The determination of tramadol hydrochloride by the fixed concentration and rate constant methods is feasible with the calibration equations obtained, but the fixed time method proves to be more applicable. PMID- 12093515 TI - Determination of codeine in urine and drug formulations using a clay-modified screen-printed carbon electrode. AB - Both flow-injection analysis and square-wave stripping voltammetry were evaluated for the determination of codeine in pharmaceutical formulations using a nontronite clay-modified screen-printed carbon electrode. Compared with a bare screen-printed carbon electrode, the nontronite clay-modified screen-printed carbon electrode exhibited a marked enhancement of the current response of codeine. A linear calibration plot was obtained over the 2.5-45 microM range (correlation coefficient=0.999) in pH 6.0 phosphate buffer solution with a detection limit of 20 nM (S/N=3) by square-wave voltammetry (SWV). While, in flow injection analysis, the linearity was over 5-120 ng range with a detection limit of 1 ng in 20 microl loop. The nontronite clay-modified screen-printed carbon electrode can be either disposable or reused since the renewal gave a good reproducible surface. Quantitative analysis was performed by the standard addition method for codeine content in both urine and commercially available drugs. PMID- 12093517 TI - Determination of the rate constants and activation energy of acetaminophen hydrolysis by capillary electrophoresis. AB - A method based on capillary electrophoresis with electrochemical detection (CE ED) was developed for the simultaneous determination of p-aminophenol and acetaminophen in the hydrolysates of acetaminophen. Effects of several important factors such as the acidity and concentration of running buffer, separation voltage, injection time, and working potential were investigated to acquire the optimum conditions. The detection electrode was a 300 microm carbon disc electrode at a working potential of +0.80 V (versus SCE). The two analytes can be well separated within 6 min in a 50 cm length fused silica capillary at a separation voltage of 18 kV in a 25 mM phosphate buffer (pH 6.5). The rate constants of acetaminophen hydrolysis in 0.5 M HCl at different temperatures were determined by monitoring the concentration changes of acetaminophen. At 70, 80, 90 and 100 degrees C, the measured rate constants of acetaminophen hydrolysis were 5.027 x 10(-3), 8.522 x 10(-3), 18.60 x 10(-3) and 32.76 x 10(-3) min(-1), respectively. The activation energy for acetaminophen hydrolysis was calculated to be 68.13 kJ mol(-1), which is in good agreement with the value in the literature. PMID- 12093518 TI - Kinetic fluorimetric determination of the antineoplastic methotrexate (MTX) in human serum. AB - A kinetic study of the oxidation of methotrexate (MTX) in acidic medium and in the presence of potassium permanganate has been made on the basis of the fluorescence-time curves. A kinetic method for the determination of MTX was developed with a range of application between 0.22 and 3.30 microM. The proposed kinetic method permits us to determine MTX in human serum and to avoid the natural fluorescence of the serum. A detection limit of 0.18 microM was calculated in the presence of ascorbic acid as activator. Only 100 s per sample is necessary for the analysis. The interference of pteridin derivatives and the rescue agent folinic acid (leucovorin) was tested. PMID- 12093519 TI - Sensitive spectrophotometric methods for the determination of amoxycillin, ciprofloxacin and piroxicam in pure and pharmaceutical formulations. AB - Two simple, sensitive and accurate spectrophotometric methods have been proposed for the determination of amoxycillin (AMX), ciprofloxacin (CPF) and piroxicam (PIR) in pure and pharmaceutical preparations. The methods are based on the measurement of absorbances of tris(o-phenanthroline) iron(II) [method A] and tris (bipyridyl) iron(II) [method B] complexes at 510 and at 522 nm, respectively. Reaction conditions have been optimized to obtain coloured complexes of higher sensitivity and longer stability. The absorbances were found to increase linearly with increase in concentrations of AMX, CPF and PIR which were corroborated by correlation coefficient values. The complexes obeyed Beer's law over the concentration range of 0.06-5.2, 0.04-7.2 and 0.2-6.5 microg ml(-1) for AMX, CPF and PIR, respectively, in method A, and of 0.05-8.5, 0.05-9.0 and 0.05-6.5 microg ml(-1) for AMX, CPF and PIR, respectively, in method B. The developed methods have been successfully applied for the determination of AMX, CPF and PIR in bulk drugs and pharmaceutical formulations. The common excipients and additives did not interfere in their determinations. The results obtained by the proposed methods have been statistically compared by means of Student t-test and by the variance ratio F-test. PMID- 12093520 TI - Spectrophotometric determination of lisinopril in tablets using 1-fluoro-2,4 dinitrobenzene reagent. AB - A spectrophotometric method for the determination of lisinopril (LN) in single and multicomponent tablets also containing hydrochlorothiazide (HCT), based on the derivatization reaction with 1-fluoro-2,4-dinitrobenzene (FDNB, Sanger reagent) is described. Aqueous solutions of LN (4.5-27.2 x 10(-5) M) react with FDNB (in acetonitrile) at pH 8.2 (borate buffer) in dark at 60 degrees C for 45 min. After acidification with HCl to decolourize 2,4-dinitrophenolate (the alkaline hydrolysis product of FDNB), the LN-derivative is measured at 356.5 or 405.5 nm (only at 405.5 nm if HCT is present). The calibration curves are linear (r>0.996 at both wavelengths) with a between days precision of slopes of 1.8 and 2.3% at 405.5 and 356.5 nm, respectively. The quantification limit is 3.49 x 10( 5) M (0.014 mg) at 405.5 nm and 5.69 x 10(-5) M (0.023 mg) at 356.5 nm. The accuracy and precision of the method were evaluated with the analysis of synthetic mixtures (Er%: 0.30-0.60 and 0.27-1.00 at 405.5 and 356.5 nm, respectively; RSD%: 0.48-0.92 and 0.35-0.51 at 405.5 and 356.5 nm, correspondingly; recovery%: 99.2-100.4 at 405.5 nm and 97.9-104.3 at 356.5 nm). Results obtained from the analysis of commercial preparations with the proposed method are in good agreement with those obtained with the official HPLC method (% relative difference 0.2-2.5%). The developed method can be used for rapid routine analysis for content uniformity, dissolution profile studies and assay of pharmaceutical preparations. PMID- 12093521 TI - LC determination of rosiglitazone in bulk and pharmaceutical formulation. AB - An isocratic reversed phase liquid chromatographic (RP-LC) method has been developed and subsequently validated for the determination of rosiglitazone and its related impurities. Separation was achieved with a Symmetry C18 column and sodium phosphate buffer (pH adjusted to 6.2):acetonitrile (50:50, v/v) as eluent, at a flow rate of 1.0 ml/min. UV detection was performed at 245 nm. The method is simple, rapid, selective and stability indicating. Indole was used as internal standard for the purpose of quantification of rosiglitazone. The described method is linear over a range of 0.45-10 microg/ml for related impurities and 180-910 microg/ml for assay of rosiglitazone. The method precision for the determination of assay and related compounds was below 1.0 and 3.6% RSD, respectively. The mean recoveries of impurities were found to be in the range of 95-102%. The percentage recoveries of Active Pharmaceutical Ingredient (API) from dosage forms ranged from 99.02 to 101.30. The method is useful in the quality control of bulk manufacturing and also in pharmaceutical formulations. PMID- 12093522 TI - Validated HPLC method for determination of sennosides A and B in senna tablets. AB - This study developed an efficient and reliable ion-pair liquid chromatographic method for quantitation of sennosides A and B in commercial senna tablets. Separation was conducted on a Hypersil C 18 column (250 x 4.6 mm, 5 microm) at a temperature of 40 degrees C, using a mixture of 0.1 M acetate buffer (pH 6.0) and acetonitrile (70:30, v/v) containing 5 mM tetrahexylammonium bromide as mobile phase. Sennosides A and B were completely separated from other constituents within 14 min. The developed method was validated. Both run-to-run repeatability (n=10) and day-to-day reproducibility (n=3) of peak area were below 0.4% RSD. Linearity of peak area was tested in the range 30-70 microg/ml (r>0.9997). Accuracy was assessed with recovery and the recoveries for sennosides A and B were 101.73+/-1.30% and 101.81+/-2.18% (n=3 x 6), respectively. Robustness of the analytical method was tested using a three-leveled Plackett-Burman design in which 11 factors were assessed with 23 experiments. Eight factors (column, concentration of ion pair reagent, % of organic modifier (acetonitrile), buffer pH, column temperature, flow rate, time constant and detection wavelength) were investigated in a specified range above and below the nominal method conditions. It was found that: (1) column and % acetonitrile affected significantly resolution and retention time, (2) column, % acetonitrile, column temperature, flow rate and time constant affected significantly the plate number of sennoside A, and (3) column and time constant affected significantly the tailing factor. PMID- 12093523 TI - SPME-LC with UV detection to study delorazepam-serum albumin interactions. AB - Solid phase microextraction coupled to high performance liquid chromatography with UV detection (SPME/LC-UV) has been employed to study the binding of delorazepam to human serum albumin (HSA) and bovine serum albumin (BSA). The procedure could also be potentially extended to the measurement of partition coefficients between a wide variety of semi- or non-volatile compounds and matrices. The method is solvent free, simple, fast, and drawbacks of the conventional analytical techniques are avoided. Moreover, the matrix did not interfere with the measurement by binding to the fibre and the amount extracted by the fibre was negligibly small; thus it did not disturb the delorazepam protein binding. PMID- 12093524 TI - A new high-performance liquid chromatography (HPLC) method for the analysis of halofantrine (HF) in pharmaceuticals. AB - A simple, rapid and stability-indicating reverse-phase high-performance liquid chromatography (HPLC) method was developed for the assay of halofantrine (HF) base, 1,3-dichloro-alpha-[2-(dibutylamino)ethyl]-6-(trifluoromethyl)-9 phenanthrenemethanol. The HPLC method was validated for precision, accuracy, selectivity and linearity and range. It was used to assay for HF base in solid dispersions. Excellent linearity was observed between HF base concentration and the peak area (R(2)=0.9998). The limit of detection was 1 ng (with a signal-to noise ratio of 2:1), and the limit of quantitation was 10 ng (with a signal-to noise ratio of 10:1). The method proved to be selective. Selectivity was validated by subjecting a stock solution of HF to acidic, basic, oxidative and thermal degradations. The peaks of the degradation products did not interfere with the peak of HF. Excipients present in the solid dispersions did not interfere with the analysis. PMID- 12093525 TI - Determination of barbituric acid, utilizing a rapid and simple colorimetric assay. AB - Barbituric acid is widely used in the manufacturing of plastics, textiles, polymers, and pharmaceuticals. Three assay methods are presented, which can utilize either aqueous or solid samples. The detection of as little as 10 microg of barbituric acid can be accomplished, either as an aqueous or solid sample, when using micro reaction tubes. The red-violet endpoint is easily discerned and results upon the formation of a violuric acid derivative. A Spot Test protocol is described which allows for a positive/negative indication of barbituric acid presence. The Spot Test is sensitive to as little as 18.75 microg/ml of barbituric acid. The construction of a Standard Curve for assaying multiple samples and over longer time periods is demonstrated, and is shown to be linear from 18.75 microg/ml to 2.25 mg/ml of barbituric acid. Spectrophotometer readings are made from an absorption peak appearing at 530 nm. The molar absorptivity of the violuric acid derivative is calculated to be 31.4 per mol/l per cm. The micro reaction tube assay will quantitate as little as 10 microg of barbituric acid through interpolation with controls containing a known amount of analyte. Inorganic salts such as NaCl, NaN(3), LiBr, and CaCO(3) do not interfere with endpoint determination. Many organic compounds (also pharmaceuticals) do not inhibit the reaction. PMID- 12093526 TI - Liquid chromatographic assay of moxidectin in human plasma for application to pharmacokinetic studies. AB - A sensitive and selective high-performance liquid chromatography (HPLC) method is presented for the analysis of moxidectin in human plasma. Solid phase extraction using Oasis HLB cartridges is used for sample preparation. The fluorescent derivative is obtained by a dehydrative reaction with trifluoroacetic anhydride and N-methylimidazole. Separation is achieved on a Bondapak C(18) reversed-phase column with a mobile phase composed of tetrahydrafuran-acetonitrile-water (40:40:20, v/v/v). Detection is by fluorescence, with excitation at 365 nm and emission at 475 nm. The retention times of moxidectin and internal standard, ivermectin are approximately 10.7 and 18.6 min, respectively. The assay is linear over the concentration range 0.2-1000 ng/ml for moxidectin in human plasma (r=0.9999, weighted by 1/concentration). Recoveries at concentrations 0.2, 400, 1000 ng/ml are 94, 75, and 71%, respectively. The analysis of quality control (QC) samples for moxidectin (0.2, 400, and 1000 ng/ml) demonstrates excellent precision with relative standard deviations of 11.9, 5.7, and 2.7%, respectively (n=6). The method is accurate with all intra- (n=6) and inter-day (n=18) mean concentrations within (5.0%) from nominal at all QC sample concentrations. Moxidectin was found to be stable after three free-thaw cycles, and with storage at -20 and -80 degrees C for 12 weeks. The method is suitable for pharmacokinetic studies of moxidectin after oral administration to humans. PMID- 12093527 TI - Determination of terbutaline sulfate and its degradation products in pharmaceutical formulations using LC. AB - There is a lack of information concerning analysis of terbutaline sulfate and quantification of its related substances particularly in the liquid dosage forms. This work aimed at developing and validating an HPLC method for determination of terbutaline sulfate and its possible degradation products, namely, 3,5 dihydroxybenzoic acid, 3,5 dihydroxybenzaldehyde and 1-(3,5-dihydroxyphenyl)-2 [(1,1-dimethylethyl) amino]-ethanone that might appear as impurities in the starting material as well as in the solid and liquid formulations. The chromatographic system used consisted a Hypersil 100 C(18,) 150 x 4.6 mm (5 microm) column, a mobile phase of ammonium acetate (0.15 M) and glacial acetic acid (pH of 4.0, 96:4 v/v) with a flow rate of 2 ml min(-1) and a UV detector set at 270 nm. The degree of linearity and the characteristic statistical parameters of the calibration curves including the limit of detection (LOD) and limit of quantitation (LOQ) were estimated for terbutaline sulfate and its degradation products. The method was found to be specific, stability indicating, accurate, precise and robust. PMID- 12093528 TI - A simple LC method with UV detection for the analysis of creatine and creatinine and its application to several creatine formulations. AB - The objective of this study was to develop a simple and sensitive LC method for the determination of creatine and creatinine in various creatine supplement formulations. The chromatographic system comprised of a LC-600 pump, SCL-6B system controller, and SPD-6AV detector (Shimadzu, Japan). The mobile phase consisted of 0.045 M ammonium sulfate in water. The chromatographic separation was achieved at ambient temperature on a Betabasic C-18 column (250 x 4.6 mm, Keystone Sci.). The flow rate was maintained at 0.75 ml/min and effluents are monitored at 205 nm. 4-(2-Aminoethyl)benzene sulfonamide was used as an internal standard (IS). This method required less than 7 min of chromatographic time. The standard curves were linear over the concentration range of 1-100 microg/ml for creatine and 2-100 microg/ml for creatinine, respectively. The relative standard deviations (RSD) for the within-day and day-to-day precision for creatine were within 1.0-4.6 and 2.2-4.7%, respectively. The RSD for the accuracy of creatine assay was in the range of 2.4-4.7%. The RSD values for the within-day precision, day-to-day precision and accuracy for creatinine validation were 1.7-4.4, 2.3-5.4 and 2.4-4.8%, respectively. This method was used to determine: (i) the creatine concentration in various marketed products; (ii) saturated solubility of various creatine salts; and (iii) stability of creatine in aqueous solution. In conclusion, a simple and sensitive LC method with UV detection was developed for the simultaneous determination of creatine and creatinine in formulations. Di creatine citrate salt showed a higher aqueous solubility (at 25 degrees C) as compared to creatine and creatine monohydrate. Some of the over-the-counter (OTC) products tested contained a very low level of creatine in contrast to their label claim. Substantial conversion of creatine into creatinine was noticed in liquid formulation. PMID- 12093529 TI - Characterization of urinary metabolites of vitamin D(3) in man under physiological conditions using liquid chromatography-tandem mass spectrometry. AB - The characterization of the urinary metabolites of vitamin D(3) in man under physiological conditions was performed using liquid chromatography-tandem mass spectrometry (LC-MS-MS). The urine specimens obtained from healthy volunteers were treated with beta-glucuronidase, purified with disposal solid-phase extraction cartridges, derivatized with a Cookson-type reagent, 4-[2-(6,7 dimethoxy-4-methyl-3-oxo-3,4-dihydroquinoxalyl)ethyl]-1,2,4-triazoline-3,5-dione, and subjected to LC-MS-MS. The derivatization was employed to increase the ionization efficiencies of the vitamin D(3) metabolites, which enabled detection of the metabolites in the picogram range. The identification of the genin parts of the metabolites was done by comparison with authentic samples based on their LC-MS-MS data. The glucuronides of 23S,25-dihydroxyvitamin D(3) and 24R,25 dihydroxyvitamin D(3) were obtained as the main metabolites from the urine in almost equal amounts. In contrast to the fact that the plasma/serum concentration of the former is much lower than that of the latter, the hydroxylation at the C 23 position was considered to be the important side-chain modification of 25(OH)D(3) to excrete the excess vitamin D(3) in man. In addition, 23S,25 dihydroxy-24-oxovitamin D(3) occurred as its glucuronide in most of the urine, which suggested that this metabolite also plays a part in the excretion of vitamin D(3) in man. PMID- 12093530 TI - Microbiological assay for azithromycin in pharmaceutical formulations. AB - The validation of a microbiological assay, applying the cylinder-plate method, for the determination of the antibiotic azithromycin is described. Using a strain of Micrococcus luteus ATCC 9341 as the test organism, azithromycin at concentrations ranging from 0.1 to 0.4 microgml(-1) could be measured in capsules and suspensions. A prospective validation of the method showed that it was linear (r=0.998), precise (RSD=1.40-capsules; RSD=1.19-powder for suspension and RSD=1.73-oral suspension) and accurate (it measured the added quantities). We conclude that the microbiological assay is satisfactory for quantitation of in vitro antibacterial activity of azithromycin. PMID- 12093531 TI - Quantification of plasma S-adenosylmethionine and S-adenosylhomocysteine as their fluorescent 1,N(6)-etheno derivatives: an adaptation of previously described methodology. AB - A simplified reversed phase HPLC system for the detection of fluorescent 1,N(6) etheno derivatives of SAM (S-adenosylmethionine) and S-adenosylhomocysteine (SAH) is described. The most important changes from the previously reported method are a shorter derivatization reaction time, the use of a solid-phase extraction resulting in an increase of the method's sensitivity, and the use of only one chromatographic system to separate SAM and SAH (in which the use of an ion pairing reagent in the mobile phase is avoided). The linearity of the method was established, and the intra-assay coefficients of variation were 10.4 and 4.7% for SAM and SAH, respectively. Normal plasma values (n=8), evaluated with the present methodology, were, for SAM and SAH, respectively, 57+/-12 and 28+/-3 nM (mean+/ SD). PMID- 12093532 TI - Fluorescent lipid probes: some properties and applications (a review). AB - Odd as it may seem, experimental challenges in lipid research are often hampered by the simplicity of the lipid structure. Since, as in protein research, mutants or overexpression of lipids are not realistic, a considerable amount of lipid research relies on the use of tagged lipid analogues. However, given the size of an average lipid molecule, special care is needed for the selection of probes, since if the size and intramolecular localization of the probe is not specifically taken into account, it may dramatically affect the properties of the lipids. The latter is particularly important in cell biological studies of lipid trafficking and sorting, where the probed lipid should resemble its natural counterpart as closely as possible. On the other hand, for biophysical applications, these considerations may be less critical. Here we provide a brief overview of the application of several lipid probes in cell biological and biophysical research, and critically analyze their validity in the various fields. PMID- 12093533 TI - Molecular and fluorescent sterol approaches to probing lysosomal membrane lipid dynamics. AB - Although the most exogenous lipids enter the cell via the LDL-receptor pathway, the mechanism(s) whereby lipids leave the lysosome for transport to intracellular sites are not clearly resolved. As shown herein, expression of sterol carrier protein-2 (SCP-2) in transfected L-cells altered lysosomal membrane lipid distribution, dynamics, and response to lipid transfer proteins. SCP-2 expression decreased the mass of cholesterol and lyso-bis-phosphatidic acid [LBPA], as well as the ratios of cholesterol/phospholipid and polyunsaturated/monounsaturated fatty acids esterified to lysosomal membrane phospholipids. Concomitantly, a fluorescent sterol transfer assay showed that SCP-2 expression decreased the initial rates of spontaneous and SCP-2-mediated sterol transfer 5.5- and 3.8 fold, respectively, from lysosomal membranes isolated from SCP-2 expressing cells as compared to controls. SCP-2, sphingomyelinase, low density lipoprotein, and high density lipoprotein directly enhanced the initial rates of sterol transfer from isolated lysosomal membranes by 50-, 12-, 4-, and 5-fold, respectively. In contrast, albumin and cholesterol esterase had no effect on lysosomal sterol transfer. Spontaneous sterol was very slow, t(1/2)>4 days, regardless of the source of the lysosomal membrane, while SCP-2 added in vitro induced formation of rapid and slowly transferable sterol pools in lysosomal membranes of control cells. In contrast, SCP-2 did not induce formation of a rapidly transferable sterol domain in lysosomal membranes isolated from SCP-2 expressing cells. These data suggest that SCP-2 expression selectively shifted the distribution of lipids (cholesterol, LBPA, esterified polyunsaturated fatty acids) away from lysosomal membranes. Furthermore, the cholesterol depleted lysosomal membrane isolated from SCP-2 expressing cells was resistant to additional direct action of SCP-2 to further enhance sterol transfer and induce rapidly transferable sterol pools in the lysosomal membrane. PMID- 12093534 TI - Fluorescent lipid probes in the study of viral membrane fusion. AB - Fluorescent lipid probes are widely used in the observation of viral membrane fusion, providing a sensitive method to study fusion mechanism(s). Due to the wealth of data concerning liposome fusion, a variety of fusion assays has been designed including fluorescent probe redistribution, fluorescence dequenching, fluorescence resonance energy transfer and photosensitized labeling. These methods can be tailored for different virus fusion assays. For instance, virions can be loaded with membrane dye which dequenches at the moment of membrane merger. This allows for continuous observation of fusion and therefore kinetic information can be acquired. In the case of cells expressing viral envelope proteins, dye redistribution studies of lipidic and water-soluble fluorophores yield information about fusion intermediates. Lipid probes can be metabolically incorporated into cell membranes, allowing observation of membrane fusion in vitro with minimal chance of flip flop, non-specific transfer and formation of microcrystals. Fluorescent lipid probes have been incorporated into liposomes and/or reconstituted viral envelopes, which provide a well-defined membrane environment for fusion to occur. Interactions of the viral fusion machinery with the membrane can be observed through the photosensitized labeling of the interacting segments of envelope proteins with a hydrophobic probe. Thus, fluorescent lipid probes provide a broad repertoire of fusion assays and powerful tools to produce precise, quantitative data in real time required for the elucidation of the complex process of viral fusion. PMID- 12093535 TI - Pyrene-labeled lipids as tools in membrane biophysics and cell biology. AB - Pyrene is one of the most frequently used lipid-linked fluorophores. Its most characteristic features are a long excited state lifetime and (local) concentration-dependent formation of excimers. Pyrene is also hydrophobic and thus does not significantly distort the conformation of the labeled lipid molecule. These characteristics make pyrene lipids well-suited for studies on a variety of biophysical phenomena like lateral diffusion, inter- or transbilayer movement of lipids and lateral organization of membranes. Pyrene lipids have also been widely employed to determine protein binding to membranes, lipid conformation and the activity of lipolytic enzymes. In cell biology, pyrene lipids are promising tools for studies on lipid trafficking and metabolism, as well as for microscopic mapping of membrane properties. The main disadvantage of pyrene lipids is the relatively large size of the fluorophore. Another disadvantage is that they require UV-excitation, which is not feasible with all microscopes. PMID- 12093536 TI - The use of fluorescent phorbol esters in studies of protein kinase C-membrane interactions. AB - The family of protein kinase C (PKC) isozymes belongs to a growing class of proteins that become active by associating with membranes containing anionic phospholipids, such as phosphatidylserine. Depending on the particular PKC isoform, this process is mediated by Ca(2+)-binding to a C2 domain and interaction of activators such as 1,2-diacyl-sn-glycerol or phorbol esters with tandem C1 domains. This cooperation between the C1 and C2 domains in inducing the association of PKC with lipid membranes provides the energy for a conformational change that consists of the release of a pseudosubstrate sequence from the active site, culminating in activation. Thus, the properties of the interactions of the C1 and C2 domains with membranes, both as isolated domains, and as modules in the full length PKC isoforms, have been the subject of intense scrutiny. Here, we review the findings of studies in which fluorescent phorbol esters have been utilized to probe the properties of the C1 domains of PKC with respect to the interaction with activators, the subsequent interaction with membranes, and the role of the activating conformational change that leads to activation. PMID- 12093537 TI - High-field electron spin resonance of spin labels in membranes. AB - High-field electron spin resonance (ESR) spectroscopy is currently undergoing rapid development. This considerably increases the versatility of spin labelling which, at conventional field strengths, is already well established as a powerful physical technique in membrane biology. Among the unique advantages offered by high-field spectroscopy, particularly for spin-labelled lipids, are sensitivity to non-axial rotation and lateral ordering, a better orientational selection, an extended application to rotational dynamics, and an enhanced sensitivity to environmental polarity. These areas are treated in some depth, along with a detailed consideration of recent developments in the investigation of transmembrane polarity profiles. PMID- 12093538 TI - Investigation on lipid asymmetry using lipid probes: Comparison between spin labeled lipids and fluorescent lipids. AB - Synthetic lipids with a nitroxide or a fluorescent probe have been extensively used during the last 30 years to determine the transmembrane diffusion of phospholipids in artificial or biological membranes. However, the relevance of data obtained with these modified lipids has sometimes been questioned. Beside possible artefacts introduced by the reporter probe, synthetic lipids used in cells often contain a short fatty acid chain in the sn-2 position, which gives them higher water solubility than naturally occurring lipids. In the present review, we have attempted to give a critical appraisal. Main strategies are recalled and important discoveries obtained with lipid probes on transmembrane lipid traffic in eukaryotic cells are briefly summarized. Examples of artefacts caused by lipid probes are given. Comparisons between data obtained by different techniques such as ESR and fluorescence allow us to emphasize the complementary character of the two approaches and more generally show the necessity to use several probes before drawing conclusions concerning endogenous lipids. In spite of these pitfalls, overall, lipid probes have provided a wealth of useful information that, to date, cannot be obtained with unlabeled lipids. PMID- 12093539 TI - Novel NMR tools to study structure and dynamics of biomembranes. AB - Nuclear magnetic resonance (NMR) studies on biomembranes have benefited greatly from introduction of magic angle spinning (MAS) NMR techniques. Improvements in MAS probe technology, combined with the higher magnetic field strength of modern instruments, enables almost liquid-like resolution of lipid resonances. The cross relaxation rates measured by nuclear Overhauser enhancement spectroscopy (NOESY) provide new insights into conformation and dynamics of lipids with atomic-scale resolution. The data reflect the tremendous motional disorder in the lipid matrix. Transfer of magnetization by spin diffusion along the proton network of lipids is of secondary relevance, even at a long NOESY mixing time of 300 ms. MAS experiments with re-coupling of anisotropic interactions, like the 13C-(1)H dipolar couplings, benefit from the excellent resolution of 13C shifts that enables assignment of the couplings to specific carbon atoms. The traditional 2H NMR experiments on deuterated lipids have higher sensitivity when conducted on oriented samples at higher magnetic field strength. A very large number of NMR parameters from lipid bilayers is now accessible, providing information about conformation and dynamics for every lipid segment. The NMR methods have the sensitivity and resolution to study lipid-protein interaction, lateral lipid organization, and the location of solvents and drugs in the lipid matrix. PMID- 12093540 TI - Fluorescence studies of lipid regular distribution in membranes. AB - This article reviews the use of fluorescent lipids and free probes in the studies of lipid regular distribution in model membranes. The first part of this article summarizes the evidence and physical properties for lipid regular distribution in pyrene-labeled phosphatidylcholine (PC)/unlabeled PC binary mixtures as revealed by the fluorescence of pyrene-labeled PC. The original and the extended hexagonal superlattice model are discussed. The second part focuses on the fluorescence studies of sterol regular distributions in membranes. The experimental evidence for sterol superlattice formation obtained from the fluorescent sterol (i.e. dehydroergosterol) and non-sterol fluorescent probes (e.g. DPH and Laurdan) are evaluated. Prospects and concerns are given with regard to the sterol regular distribution. The third part deals briefly with the evidence for polar headgroup superlattices. The emphasis of this article is placed on the new concept that membrane properties and activities, including the activities of surface acting enzymes, drug partitioning, and membrane free volume, are fine-tuned by minute changes in the concentration of bulky lipids (e.g. sterols and pyrene-containing acyl chains) in the vicinities of the critical mole fractions for superlattice formation. PMID- 12093542 TI - 11th Association of European Psychiatrists Congress. Stockholm, Sweden, 4-8 May 2002. AEP CME course programme. PMID- 12093541 TI - Fluorescent and photoactivable probes in depth-dependent analysis of membranes. AB - This report summarizes our efforts towards depth-dependent analysis of membranes by design of suitable fluorescent and photoactivable lipid probes, which can be incorporated into membranes. The objective of depth-dependent analysis has been two fold, one to obtain information on lipid domains and other on transmembrane domains of membrane-bound proteins. In view of increasing importance of lipid rafts and other localized domain and limited success in case of structure determination of membrane-bound proteins vis-a-vis their soluble counterparts, it is tempting to rapidly attach fluorescent or photoactivable probes to lipids to get a probes where relatively little attention is paid to design of such probes. We have shown here how careful design of such probes is required to immobilize such probes in membranes for effective depth-dependent analysis of membranes. An effective design has become important when identification of putative transmembrane domains predicted primarily from the genome data based on hydropathy plots, often needs confirmation by contemporary methodology. PMID- 12093580 TI - Healthy people 2010--leading health indicators for women. AB - To examine the health status of women in relationship to the targets for the Healthy People 2010 ten Leading Health Indicators (LHIs), this paper compares females and males, as well as females by age groups. The data presented in this study were collected from publicly available sources, including Healthy People 2010 publications and websites, as well as Health US 2000. In addition, the lead agency Department of Health and Human Services workgroup coordinators provided clarifying information. Women were found to have better health status than men on half of the LHIs. When examined by age, data for the LHIs provide additional insights on where to target interventions to improve women's health. For adolescent females, the biggest challenges to meet the year 2010 targets are in reducing tobacco use, obesity, motor vehicle deaths, and homicides. For adult females, the greatest public health challenges are in increasing physical activity, reducing obesity, and obtaining immunizations. PMID- 12093581 TI - Trends in women's health services by type of physician seen: data from the 1985 and 1997-98 NAMCS. AB - As managed care enrollment has increased, controversy has arisen about the role of internists (IM), family physicians (FP), and obstetrician/gynecologists (ob/gyns) in the provision of women's health care. Efforts to improve training in women's health needs have also increased. Yet it is unclear how these trends have affected practice. We used the National Ambulatory Medical Care Survey (NAMCS), a nationally representative sample of office-based medical visits, to examine by physician specialty a) trends in the proportion of visits for women's health care and b) the content of nonillness care. Between 1985 and 1997-98, market share of reproductive health services increased for IMs (e.g., from 3.7% to 10.5% of contraceptive visits, p <.05) and decreased for FPs (from 30.5% to 20.5% for contraceptive visits, p <.05). Ob/Gyns increased their share of women's health care visits, with reproductive health visits increasing from 56.2% to 65.9% (p <.0001). The trend in hormone replacement therapy visits differed, with nonsignificant gains in market share for IMs and decreases for ob/gyns. Nonillness care (1997-98 data only) differed predictably by specialty, with IMs and FPs more often providing cholesterol screening while ob/gyns more often provided reproductive health services. Compared with IMs and FPs, ob/gyns were more likely to counsel women on reproductive health topics and equally likely to counsel on general health topics, but additional time spent in counseling was lower. Specialty differences in the provision of women's health services continue, though the scope of care provided by IMs has broadened. Still, women are unlikely to obtain a full range of preventive services in a single nonillness visit. Ensuring adequate coordination among physicians providing primary care to women continues to be a critical concern. PMID- 12093582 TI - Optimal health and well-being for women: definitions and strategies derived from focus groups of women. AB - Women participating in focus groups were asked how they define health and well being, and what strategies they would suggest for health optimization. Women defined health and well-being largely in terms of relationships. Their strategies for improving health involved enhancement of the quality of relationships with families, partners, and community. These proposed strategies included: creating a context for resilience; valuing and nurturing children, parents, and families; promoting interpersonal connections and community; realizing equality for women; and cultivating relational values. These perspectives can inform clinicians and health policy. PMID- 12093583 TI - The mismatch between urban women's preferences for and experiences with primary care. AB - Socioeconomic disparities in women's primary care experiences have been described previously. To better understand whether these disparities reflect personal preferences for primary care, rather than insurance or other access barriers, we conducted a telephone survey of a community-based sample of 1,205 women in Washington, DC. The study found that women of lower socioeconomic status had poorer primary care experiences compared with higher income counterparts, despite similarly high preferences for primary care. The poorer primary care experiences of lower socioeconomic status women were attenuated by better access to primary care. Differences in primary care attainment are not solely a matter of personal preferences; rather, they appear to be more strongly related to barriers to obtaining care. PMID- 12093584 TI - Are women worrying about heart disease? AB - Women at risk for coronary artery disease (CAD) may not be worried about the disease. In this study, demographic, CAD-risk, and CAD-worry data collected from Durham Veterans' Affairs Medical Center women veterans were examined using bivariate and multivariate analysis with worry as the outcome. Excluding CAD patients (N = 64) and incomplete data (N = 17) of 328 women, 42% worried about CAD. Younger age, single marital status, obesity, family history, and hyperlipidemia were associated with worry. Of women with up to three risk factors, fewer than half worried about CAD. Higher-risk women were unconcerned about CAD. This could undermine prevention efforts. PMID- 12093585 TI - Menopausal experiences and hormone replacement therapy use among women with physical impairments. AB - This study describes the menopausal experiences of women with physical disabilities and their knowledge and use of hormone replacement therapy (HRT). 166 women completed a mailed survey of demographic and disability characteristics, health history, menopausal information, and HRT knowledge. The typical respondent was a 52-year-old, Anglo, college-educated woman with a neuromuscular disorder. Half of the menopausal women were taking HRT, and 47% had a hysterectomy. HRT use was significantly associated with hysterectomy status, amount of HRT information received from providers, and regular gynecologic examinations. Effective interventions are needed to assist these women in making key health decisions during menopause. PMID- 12093588 TI - Striatal volume changes in the rat following long-term administration of typical and atypical antipsychotic drugs. AB - Striatal enlargement has been consistently reported in schizophrenics receiving chronic neuroleptic treatment although the results following atypical antipsychotic treatment have been equivocal. In order to disentangle patient illness from a possible drug effect on brain structure, young adult rats were administered either haloperidol, risperidone, clozapine, olanzapine, or vehicle daily for four or eight months via drinking water. Significant increases in caudate-putamen volumes were seen in animals receiving either haloperidol or clozapine when compared with control animals following eight months of drug administration. Conversely, olanzapine-treated animals showed significant decreases in caudate-putamen volumes when compared with control animals after eight months of drug. Thus, converging evidence indicates that the neuroplastic response of the striatum following neuroleptic exposure causes volumetric increases, whereas atypical antipsychotics affect the basal ganglia differentially. The current data suggests that such differential responses may be due to both the pharmacological properties and the relative doses of the atypical agents. PMID- 12093587 TI - Downregulation of BDNF mRNA in the hippocampal dentate gyrus after re-exposure to cues previously associated with footshock. AB - This study examined the effects of footshock stress and re-exposure to cues previously associated with footshock on expression of brain-derived neurotrophic factor (BDNF) mRNA in the hippocampus of male rats. Exposure to twenty 0.5-s 0.4 mA footshocks co-terminating with 70 dB, 5-s long pure tones over 60 min decreased dentate gyrus BDNF mRNA by 21.5%. Baseline BDNF mRNA levels returned to normal by two days after footshock exposure. Re-exposure for 60 min to the chamber and tones previously paired with 0.4 mA footshock decreased BDNF mRNA by 12%. Re-exposure to the conditioning chamber and tones previously paired with 0.6 mA footshock over 60 min decreased BDNF mRNA by 20.8%. The data suggest that psychological, as well as unconditioned physical stress, can decrease hippocampal BDNF mRNA. Possible implications for stress-related and other neuropsychiatric disorders associated with deficits in hippocampal function and volume, such as depression, post-traumatic stress disorder, and Alzheimer's Disease, are discussed. PMID- 12093589 TI - Chronic antipsychotic drug treatment induces long-lasting expression of fos and jun family genes and activator protein 1 complex in the rat prefrontal cortex. AB - We have characterized the effects of chronic clozapine and haloperidol treatments on the expression of fos (c-fos, fosB, fra-2) and jun (c-jun, junB, junD) family genes in the rat forebrain. The effects of chronic (17d) clozapine and haloperidol on mRNA expression were determined two hours, 24 hours, and six days after the last drug injection, and the DNA-binding activity of the activator protein-1 (AP-1) complex was studied after washout periods of 24 hours and six days. Chronic clozapine treatment with a 6 d washout period induced the expression of several fos and jun family genes in cortical regions, including the prefrontal cortex (PFC), and in the caudate putamen and nucleus accumbens. Moreover, the DNA-binding activity of the AP-1 complex was greatly increased in the anterior cingulate cortex-PFC in mobility shift assays already after 24 h, and remained increased after a 6d washout period. Chronic administration of haloperidol upregulated fos and jun family mRNA expression that was detectable 24 h and 6 d after cessation of the treatment mainly in the cortex. However, the DNA binding activity of the AP-1 complex was not altered in the anterior cingulate cortex-PFC by chronic haloperidol administration at any of the time points studied. Thus, chronic treatments with clozapine and haloperidol induce a long lasting enhancement of fos and jun family transcription factors that continues for several days after the cessation of the treatments in the cortex. These lasting effects might represent events that are potentially involved in the mechanisms of antipsychotic drug action. PMID- 12093590 TI - Acute handling stress modulates methylphenidate-induced catecholamine overflow in the medial prefrontal cortex. AB - Although stress is an extensively investigated phenomenon, the effects of specific stressors on the pharmacologic activity of routinely administered drugs are less well characterized. We designed the present study to investigate the effect of handling stress on catecholaminergic responsivity following an acute methylphenidate (MP, Ritalin) challenge in the medial prefrontal cortex (mPFC). Norepinephrine (NE) and dopamine (DA) levels were simultaneously measured in 15 min samples of PFC dialysate using HPLC coupled with electrochemical detection. Sprague-Dawley rats were handled for 15 min, which produced an increase from basal extracellular DA and NE levels. Handling stress attenuates the DA response when administered 2 h prior to IP MP, whereas handling stress enhances the DA response when administered simultaneously with IG MP. These findings suggest that persistent alterations in mesocorticolimbic DA-ergic activity are induced by a short exposure to restraint stress as evidenced by the altered response to MP challenge. PMID- 12093591 TI - Aggression escalated by social instigation or by discontinuation of reinforcement ("frustration") in mice: inhibition by anpirtoline: a 5-HT1B receptor agonist. AB - Experiments with social instigation or the omission of scheduled reinforcement show that serotonergic mechanisms may be involved in escalated aggression in animals. 5-HT1B receptor agonists have anti-aggressive effects in individuals who show moderate as well as high levels of aggression. The present study compared the effects of the 5-HT1B agonist anpirtoline (0.125-1.5 mg/kg) on (1) species typical aggressive behavior in male mice, (2) aggression "instigated" or primed by prior exposure to the opponent, and (3) aggression heightened by "frustration" caused by omission of scheduled reinforcement. The effects of anpirtoline on species-typical behavior were also assessed after pretreatment with the 5-HT1B/1D receptor antagonist GR127935 (10 mg/kg). Anpirtoline, like other 5-HT1B agonists (CP-94,253, zolmitriptan), decreased both instigated and frustration-heightened aggression, while motor behavior was unaffected. The aggression-inhibiting effects of anpirtoline were blocked by pretreatment with GR127935. The current results indicate that the 5-HT(1B) receptor is critically involved in the modulation of escalated aggression. PMID- 12093592 TI - Differential effects of caffeine on dopamine and acetylcholine transmission in brain areas of drug-naive and caffeine-pretreated rats. AB - The effects of caffeine on extracellular dopamine and acetylcholine have been studied in freely moving rats implanted with concentric microdialysis probes in the nucleus accumbens shell and core and in the medial prefrontal cortex. Intravenous administration of caffeine (0.25, 0.5, 1.0, 2.5 and 5.0 mg/kg) dose dependently increased dopamine and acetylcholine dialysate concentrations in the medial prefrontal cortex, while it did not affect dialysate dopamine in the shell and core of the nucleus accumbens. Intraperitoneal administration of caffeine (1.5, 3, 10, 30 mg/kg) also failed to affect DA in the shell and core of the nucleus accumbens. Such effects were duplicated by intravenous administration of DPCPX, a selective antagonist of adenosine A1 receptors, and of SCH 58261, an antagonist of A2a receptors. The effect of caffeine on prefrontal dopamine and acetylcholine transmission was also studied in rats chronically administered with caffeine (25 mg/kg, twice a day for seven days). At the end of this treatment rats became tolerant to the locomotor stimulating effects of a dose of 1 and 2.5 mg/kg i.v. of caffeine; these doses, however, still increased dialysate acetylcholine but did not affect dopamine in the prefrontal cortex. Therefore, in rats made tolerant to the locomotor stimulant effects of caffeine, tolerance developed to the dopamine stimulant but not to the acetylcholine stimulant effect of caffeine in the prefrontal cortex. The lack of acute stimulation of dopamine release in the nucleus accumbens shell by caffeine is relevant to the issue of its addictive properties and of the role of DA in drug- and substance-addiction. On the other hand, the dissociation between tolerance to the locomotor effects of caffeine and stimulation of acetylcholine release in the prefrontal cortex suggests that this effect might be correlated to the arousing effects of caffeine as distinct from its locomotor stimulant properties. PMID- 12093593 TI - Brain penetrance, receptor occupancy and antistress in vivo efficacy of a small molecule corticotropin releasing factor type I receptor selective antagonist. AB - The present studies were designed to evaluate the competitive binding properties and functional effects of a novel nonpeptide CRF1 receptor antagonist, R121919. R121919 administered in doses of 0.63 to 20 mg/kg p.o. 60 min pretest in Wistar rats dose dependently attenuated the swim stress-induced anxiogenic-like behavior in the elevated plus-maze model of anxiety. Moreover, receptor autoradiography revealed that R121919 dose-dependently occupied brain CRF1 receptors in subjects tested in the plus-maze experiment. Orally administered doses of up to 20 mg/kg R121919 also blunted basal and swim stress-induced pituitary-adrenocortical activation, produced additional anxiolytic-like behavioral actions in the defensive withdrawal and defensive burying paradigms, and functionally antagonized the locomotor stimulatory properties of exogenously administered CRF. Taken together, these results suggest that the anxiolytic-like efficacy of R121919 in attenuating the stress-, novelty-, shock-, and CRF-induced increases in behavioral arousal is correlated with competitive blockade of central CRF1 receptors. PMID- 12093594 TI - Conditioned opioid withdrawal decreases nociceptin/orphanin FQ levels in the frontal cortex and olfactory tubercle. AB - Clinical evidence suggests that individuals experiencing drug withdrawal can become conditioned to environmental situations, whereby previously neutral stimuli can produce symptoms of withdrawal. It is believed that this "conditioned withdrawal" can have motivational significance, but the neurobiological basis for conditioned withdrawal is unknown. The goal of this study was to determine adaptations in endogenous opioid systems that may be responsible for expression of conditioned withdrawal. Opioid-dependent rats trained to lever press for food were exposed to tone and scent cues in the presence of naloxone or saline. Naloxone but not saline predictably suppressed responding for food. One month later and in a post-dependent state, all rats again were exposed to the cues but not naloxone. The conditioned cues alone suppressed responding for food in the rats previously paired with naloxone, but no suppression was seen in rats previously paired with saline. Radioimmunoassay (RIA) analysis for nociceptin/orphanin FQ (nociceptin), met-enkephalin-Arg-Phe (MEAP), and dynorphin A (dyn A) was performed from dissections of various brain regions of the rats undergoing conditioned withdrawal. Significant reductions in nociceptin peptide levels were seen in the frontal cortex and olfactory tubercle of these rats. Unconditioned opioid withdrawal and unconditioned footshock stress produced different patterns of opioid peptide regulation in separate groups of rats. These results shed light on adaptations of endogenous opioid systems to conditioned cues, stress, and withdrawal, all factors that play a role in motivating drug intake. PMID- 12093595 TI - Peculiar vulnerability to nicotine oral self-administration in mice during early adolescence. AB - A "gateway" function toward substance abuse has been suggested for early tobacco smoking. Nicotine actually represents an easily available drug for human adolescents, who are very likely to use a number of different psychoactive agents. Surprisingly, the psychobiological factors involved in this age-related willingness have been poorly investigated. In Experiment 1, nicotine consumption was studied in outbred CD-1 mice during Early (postnatal day (pnd) 24 to 35), Middle (pnd 37 to 48) or Late (pnd 50 to 61) adolescence, in an oral self administration paradigm. During the drinking session (2 h/day), animals had free choice between either tap water or a nicotine solution (10 mg/l). After a 6-day period, a fading study was carried out, in which nicotine concentration was reduced to 7 mg/l (days 7-9) and 5 mg/l (days 10-12), to assess whether animals would compensate by increasing their intake from the nicotine solution. In Experiment 2, psychopharmacological effects on locomotion induced by the nicotine solution (0, 10, 30 mg/l) during the 1-h drinking session were assessed in Early and Late adolescent mice. In Experiment 1, Early adolescents expressed a marked and stable preference for the nicotine solution, showing a daily nicotine intake of 1.15 +/- 0.04 mg/kg. Middle adolescents did not show any preference for either bottle, whereas a tendency toward avoidance for the nicotine solution was found for Late adolescents. In the fading study, Early adolescents were the only group to show increased consumption from the nicotine bottle as far as nicotine concentration was reduced. A time-course analysis of plasma levels of cotinine (the principal biomarker of nicotine consumption) revealed some pharmacokinetic differences between the three age-groups. In Experiment 2, drinking from a nicotine solution produced a prominent hyperactivity in Early adolescents, whereas a quite opposite profile was associated with older subjects. In summary, even if a role for taste factors cannot be completely ruled out, a peculiar spontaneous drive toward oral nicotine consumption, as well as a nicotine-induced arousal, is specific to Early adolescence in mice. The present animal model might be useful to investigate psychobiological determinants involved in early tobacco smoking in human adolescents PMID- 12093596 TI - Spontaneous novelty seeking and amphetamine-induced conditioning and sensitization in adult mice: evidence of dissociation as a function of age at weaning. AB - Individual differences in coping with novelty and in the response to psychoactive drugs have been related to early life events, such as the age of weaning. Outbred CD-1 mice underwent a precocious (postnatal day (pnd) 15, Wean-15 group), regular (pnd 21, Wean-21 group), or delayed (pnd 27, Wean-27 group) weaning, and were tested as adults (pnd > 60). In Experiment 1, animals underwent a treatment history with d-amphetamine (AMPH 0, 1, or 5 mg/kg once/day for three days) in a familiar environment. On testing day, mice were challenged with SAL or a standard 1 mg/kg AMPH dose (to evaluate acute drug effects and sensitization), and placed in the familiar environment. As expected, regular Wean-21 animals showed an AMPH induced hyperactivity and a profile of conditioned locomotion, whereas the same dosage failed to induce any change in Wean-15 and Wean-27 groups. Levels of spontaneous novelty seeking were particularly elevated for Wean-27 mice, when compared with the other weaning groups. In Experiment 2, pairing of AMPH administration (0, 1, 2, 3.3, or 5 mg/kg once/day for three days) with a distinct environment produced a classical conditioned place preference. The magnitude of the preference profile was significantly more marked for Wean-15 mice, when compared with the other two weaning groups. Both experiments also provided evidence that the development of sensitization was particularly evident in Wean 27 mice. In summary, delayed weaning was associated in adult mice with both elevated levels of novelty seeking and increased sensitization to drug effects. Conversely, animals weaned precociously were much more responsive to AMPH-induced incentive conditioning. These results appear relevant to the issue of early experiences as possible risk factors for a number of psychiatric disorders in humans, including the abuse of drugs. PMID- 12093597 TI - Effects of acute and chronic reboxetine treatment on stress-induced monoamine efflux in the rat frontal cortex. AB - Reboxetine is a selective noradrenergic reuptake inhibitor that displays an antidepressant profile in both animal tests and in clinical trials. The present study examined the ability of reboxetine to alter stress-induced increases in norepinephrine, serotonin and dopamine efflux in the frontal cortex in awake behaving rats. Acute systemic administration of reboxetine (0.3-20.0 mg/kg) dose dependently increased extracellular norepinephrine in the frontal cortex while having no effect on extracellular serotonin. At 20 mg/kg, reboxetine also increased extracellular dopamine. Application of a 20-min tailpinch stress increased extracellular norepinephrine. This effect was greatly potentiated in rats pretreated with reboxetine. Tailpinch did not elicit increases in dopamine in saline treated animals but this stimulus increased dopamine levels following reboxetine pretreatment. Furthermore, chronic administration of reboxetine for 14 days resulted in elevated basal concentrations of extracellular norepinephrine and dopamine and a greater net increase of extracellular norepinephrine and dopamine, but not serotonin, in response to tailpinch compared with vehicle control animals. Taken together, these data support the view that the noradrenergic and dopaminergic systems are modified by reboxetine treatment and may be important factors in the mechanism of action of antidepressant compounds. PMID- 12093598 TI - Dopamine D2 and D3 receptor occupancy in normal humans treated with the antipsychotic drug aripiprazole (OPC 14597): a study using positron emission tomography and [11C]raclopride. AB - Aripiprazole (OPC 14597) is an antipsychotic drug that has high affinity for dopamine D2 and D3 receptors and the dopamine autoreceptor. It is being developed for treatment of patients with schizophrenia. The purpose of this study was to determine whether a dose response following graduated doses of aripiprazole could be quantified and correlated with its occupancy of the D2 and D3 dopamine receptors in the brain of living humans. Dopamine D2 and D3 receptor occupancy in fifteen normal male human brains was measured using positron emission tomography (PET) with [11C]raclopride. PET studies were performed before and after two weeks of administration of aripiprazole. The dopamine D2 receptor occupancy was quantified with two kinetic modeling methods without using a blood input function. Administration of aripiprazole for 14 days resulted in a dose-dependent receptor occupancy between 40 - 95% after the administration of 0.5mg, 1 mg, 2 mg, 10 mg, and 30 mg per day. These results suggest that an adequate occupancy can be obtained, and this may be useful to predict an appropriate therapeutic dose for an individual patient. Interestingly, even at striatal D2 receptor occupancy values above 90%, which occurred with the higher doses, extrapyramidal side effects (EPS) were not observed. This underlines aripiprazole's unique mechanism of action as a partial dopamine receptor agonist, which might become a novel principle in the treatment of schizophrenia. PMID- 12093599 TI - Effects of alprazolam on driving ability, memory functioning and psychomotor performance: a randomized, placebo-controlled study. AB - Alprazolam is prescribed for the treatment of anxiety and panic disorder. Most users are presumably involved in daily activities such as driving. However, the effects of alprazolam on driving ability have never been investigated. This study was conducted to determine the effects of alprazolam (1 mg) on driving ability, memory and psychomotor performance. Twenty healthy volunteers participated in a randomized, double-blind, placebo-controlled crossover study. One hour after oral administration, subjects performed a standardized driving test on a primary highway during normal traffic. They were instructed to drive with a constant speed (90 km/h) while maintaining a steady lateral position within the right traffic lane. Primary performance measures were the Standard Deviation of Lateral Position (SDLP) and the Standard Deviation of Speed (SDS). After the driving test, subjective driving quality, mental effort, and mental activation during driving were assessed. A laboratory test battery was performed 2.5 h after treatment administration, comprising the Sternberg Memory Scanning Test, a Continuous Tracking Test, and a Divided Attention Test. Relative to placebo, alprazolam caused serious driving impairment, as expressed by a significantly increased SDLP (F(1,19) = 97.3, p <.0001) and SDS (F(1,19) = 30.4, p <.0001). This was confirmed by subjective assessments showing significantly impaired driving quality (F(1,19) = 16.4, p <.001), decreased alertness (F(1,19) = 43.4, p <.0001), decreased mental activation (F(1,19) = 5.7, p <.03) and increased mental effort during driving (F(1,19) = 26.4, p <.0001). Furthermore, alprazolam significantly impaired performance on the laboratory tests. In conclusion, alprazolam users must be warned not to drive an automobile or operate potentially dangerous machinery. PMID- 12093600 TI - Smoking modulates neuroendocrine responses to ipsapirone in patients with panic disorder. AB - Reduced 5-HT1A-receptor responsiveness has been reported in patients with panic disorder(PD) and/or agoraphobia (PDA). Although many of these patients are regular smokers, it has not been examined whether psychological or neurobiological effects induced by the selective 5-HT1A-receptor agonist, ipsapirone, are affected by the smoking status of the patients. In order to clarify this question neuroendocrine challenges with oral doses of ipsapirone (0.3 mg/kg) and placebo were performed in 39 patients with PDA, and results were compared between patients who smoked (>10 cigarettes per day, n = 17) and patients who had been non-smokers for at least two years (n = 22). Patients who were smokers (but did not smoke during the challenge procedure) had significantly reduced baseline concentrations of cortisol and a significantly lower body temperature. In comparison to placebo, administration of ipsapirone was associated with significant increases of various psychological symptoms and plasma cortisol concentrations. The subgroup of PD patients who were smokers showed significantly higher cortisol responses to ipsapirone than non-smokers. In conclusion, smoking status has to be taken into account when assessing the responsiveness of 5-HT1A receptors in patients with psychiatric disorders. The prevention of smoking during challenge sessions might not be the ideal approach in heavy smokers, since sudden abstinence from smoking is likely to affect neurobiological and possibly psychological responses to ipsapirone. PMID- 12093601 TI - Chronic effects of Brahmi (Bacopa monnieri) on human memory. AB - A study is reported on the effects of Brahmi (Bacopa monniera) on human memory. Seventy-six adults aged between 40 and 65 years took part in a double-blind randomized, placebo control study in which various memory functions were tested and levels of anxiety measured. There were three testing sessions: one prior to the trial, one after three months on the trial, and one six weeks after the completion of the trial. The results show a significant effect of the Brahmi on a test for the retention of new information. Follow-up tests showed that the rate of learning was unaffected, suggesting that Brahmi decreases the rate of forgetting of newly acquired information. Tasks assessing attention, verbal and visual short-term memory and the retrieval of pre-experimental knowledge were unaffected. Questionnaire measures of everyday memory function and anxiety levels were also unaffected. PMID- 12093602 TI - Scopolamine but not lorazepam modulates face repetition priming: a psychopharmacological fMRI study. AB - Repetition priming is a basic form of learning associated with decreased neuronal responses following stimulus repetition. In this experiment, we address cholinergic and GABAergic modulation of repetition priming in a face recognition paradigm. In experiment 1, we used event-related functional magnetic resonance imaging (fMRI) in combination with pharmacological challenge where participants were given placebo, lorazepam (2mg po), or scopolamine (0.4 mg IV) prior to study. Behavioral data showed intact priming for famous faces in the placebo and lorazepam group but impaired priming following scopolamine. In within-group analyses, a right fusiform region showed a fame by repetition interaction characterized by a response decrease to repetition of famous faces and a response enhancement to repetition of unfamous faces in the placebo group. In subjects treated with lorazepam, a main effect of repetition, driven by response decreases to repetition of famous faces, was seen in this right fusiform region. No significant repetition effects were found after scopolamine. In experiment 2, we further investigated behaviorally the cholinergic impairment of repetition priming. Participants were given either placebo or scopolamine (0.4 mg IV) after study. Behavioral data showed intact priming for famous faces in the placebo and scopolamine group. The results suggest that scopolamine but not lorazepam impair repetition priming for famous faces in a face recognition paradigm. These cholinergic impairments are likely to reflect interference with acquisition processes during study that may co-occur with a modulation of right fusiform decreases to repetition of famous faces. PMID- 12093603 TI - Effects of risperidone on the peripheral noradrenegic system in patients with schizophrenia: a comparison with clozapine and placebo. AB - Risperidone is an atypical antipsychotic drug that increases plasma norepinephrine (NE) levels, but the mechanism behind this effect is unclear. We measured arterial plasma levels of NE and other catechols during intravenous infusion of tritium-labeled NE (3H-NE) in risperidone-treated patients and compared their data with those from patients treated with clozapine or placebo. NE levels in risperidone patients were significantly higher than in placebo patients, but lower than in clozapine patients. Neither drug, however, had significant effect on plasma levels of the main neuronal metabolite of NE, dihydroxyphenylglycol (DHPG), suggesting that adrenoceptors blockade alone would not explain the NE findings. The rate of release of endogenous NE into the bloodstream (spillover) was elevated in both risperidone and clozapine patients in a manner that paralleled their NE levels; the NE clearance in both groups did not differ from placebo. Following 3H-NE infusion in risperidone-treated individuals, production of 3H-DHPG was normal, as it was in the clozapine group, suggesting that risperidone does not impede neuronal uptake or intraneuronal metabolism of NE by monoamine oxidase. Our data suggest that both risperidone and clozapine elevate plasma NE levels via enhanced neurotransmitter spillover, with risperidone producing a smaller effect. PMID- 12093604 TI - Serotonin 1A receptor activation and hypothermia in humans: lack of evidence for a presynaptic mediation. AB - The hypothermia produced by 5-HT1A agonists had initially been claimed to be caused by the activation of cell body 5-HT1A autoreceptors resulting in decreased 5-HT transmission in laboratory animals. In order to address this issue in humans, 12 healthy volunteers underwent a dietary tryptophan depletion paradigm to decrease 5-HT availability, under double-blind conditions, during which body temperature was monitored following oral administration of the 5-HT1A agonist buspirone (30 mg). In addition, plasma prolactin and growth hormone evaluations, two responses that are mediated via the direct activation of postsynaptic 5-HT1A receptors, were determined. The hypothesis was that if responses are mediated by decreased transmission at postsynaptic 5-HT1A receptors, resulting from dampened 5-HT release as a consequence of 5-HT1A autoreceptors activation, then responses to the exogenous 5-HT1A agonist should be attenuated when 5-HT availability has been markedly decreased beforehand. Buspirone produced the same significant increase in prolactin and growth hormone in the tryptophan-depleted state as in the control condition. Similarly, the degree of hypothermia produced by buspirone was not significantly different in the two experimental conditions. In conclusion, these results strongly suggest that the hypothermia and the increases in prolactin and growth hormone produced by buspirone are attributable to the enhanced activation of postsynaptic 5-HT1A receptors, and not to a decrease in 5 HT transmission resulting from the activation of the 5-HT1A cell body autoreceptors on 5-HT neurons. PMID- 12093605 TI - Glucocorticoid hormones and early brain development in schizophrenia. AB - While genetic factors play a role in the etiology of schizophrenia, environmental factors contribute to the etiology of the disorder as well. If a woman is exposed during the second trimester of gestation to stressors such as the death of a loved one, influenza infection, or natural and man-made disasters, her baby has an increased risk of developing schizophrenia later in life. A common mechanism may mediate the effects of these diverse risk factors: a stress response during a restricted period of gestation may affect fetal brain development and, given the appropriate genetic predisposition, result in schizophrenia in adult life. Findings in animals exposed to prenatal stress strengthen the hypothesis that exposure to high glucocorticoid levels and/or other components of the stress response increases the risk of schizophrenia. These clinical and preclinical studies could be used to develop hypotheses that could then be tested in patients with schizophrenia. PMID- 12093607 TI - Adolescent risk and vulnerability: overview. PMID- 12093608 TI - Perceptions of risk and vulnerability. PMID- 12093609 TI - Vulnerability, risk, and protection. PMID- 12093610 TI - Modeling the payoffs of interventions to reduce adolescent vulnerability. PMID- 12093611 TI - Adolescent vulnerability: measurement and priority setting. PMID- 12093613 TI - Mangiferin protects the streptozotocin-induced oxidative damage to cardiac and renal tissues in rats. AB - The role of oxidative stress in streptozotocin (STZ)-induced toxicity and its prevention by a xanthone glucoside, mangiferin was investigated. To induce diabetes mellitus, adult male Wistar rats were injected STZ intravenously at 55 mg/kg body weight. The effect of mangiferin (10 and 20 mg/kg, i.p., 28 days) was investigated in STZ-induced diabetic male rats. Insulin-treated rats (6 U/kg, i.p., 28 days) served as positive control. Diabetic rats given normal saline served as negative control. Normal rats that neither received STZ nor drugs served as normal control. On day 28, the diabetic rats showed significant increase in serum creatine phosphokinase (CPK) and total glycosylated haemoglobin. Kidney revealed tubular degeneration and decreased levels of superoxide dismutase (SOD) and catalase (CAT) with an elevation of malonaldehyde (MDA). Cardiac SOD, CAT and lipid peroxidation were significantly increased. Histopathological findings revealed cardiac hypertrophy with haemorrhages. Analysis of erythrocyte revealed significantly elevated levels of MDA with insignificant decrease in CAT and SOD. Repeated intraperitoneal injections of mangiferin (10 and 20 mg/kg) and insulin (6 U/kg) controlled STZ-induced lipid peroxidation and significantly protected the animals against cardiac as well as renal damage. From the study, it may be concluded that oxidative stress appears to play a major role in STZ-induced cardiac and renal toxicity as is evident from significant inhibition of antioxidant defence mechanism in renal tissue or a compensatory increase in antioxidant defence mechanism in cardiac tissue. Intraperitoneal administration of mangiferin exhibited significant decrease in glycosylated haemoglobin and CPK levels along with the amelioration of oxidative stress that was comparable to insulin treatment. PMID- 12093614 TI - Interactions of fluorochemicals with rat liver fatty acid-binding protein. AB - Liver-fatty acid binding protein (L-FABP) is an abundant intracellular lipid carrier protein. The hypothesis that perfluorooctanesulfonate (PFOS), perfluorooctanoate (PFOA), and certain related perfluorooctanesulfonamide-based fluorochemicals (PFOSAs) can interfere with the binding affinity of L-FABP for fatty acids was tested. The relative effectiveness of PFOA, PFOS, N ethylperfluorooctanesulfonamide (N-EtFOSA), N-ethylperfluorooctanesulfonamido ethanol (N-EtFOSE), and of the strong peroxisome proliferator Wyeth-14643 (WY) to inhibit 11-(5-dimethylaminonapthalenesulphonyl)-undecanoic acid (DAUDA) binding to-L-FABP was determined. The dissociation constant (Kd) of the DAUDA-L-FABP complex was 0.47 nM. PFOS exhibited the highest level of inhibition of DAUDA-L FABP binding in the competitive binding assays, followed by N-EtFOSA, WY, and, with equal IC(50)s, N-EtFOSE and PFOA. The in vitro data presented in this study support the hypothesis that these fluorochemicals may interfere with the binding of fatty acids or other endogenous ligands to L-FABP. Furthermore, this work provides evidence to support the hypothesis that displacement of endogenous ligands from L-FABP may contribute to toxicity in rodents fed these fluorochemicals. PMID- 12093615 TI - In vitro induction of cytochrome P450 2B1- and 3A1-mRNA and enzyme immunostaining in cryopreserved precision-cut rat liver slices. AB - With the exception of cytochrome P450 (CYP) 1A1 and its mRNA, in vitro induction of other CYP forms has not been demonstrated in cryopreserved liver slices until now. Therefore precision-cut rat liver slices were cultured after cryopreservation and thawing in William's medium E for up to 24 h in the presence of inducers to demonstrate CYP2B1- and CYP3A1-mRNA induction. CYP-mRNA expression was determined by competitive RT-PCR. Exposure to 100 microM phenobarbital caused a more than 20-fold increase in CYP2B1-mRNA expression within 24 h, reaching concentrations comparable with those of PB-exposed fresh rat liver slices. Exposure to 1 microM pregnenolone 16 alpha-carbonitrile enhanced CYP3A1-mRNA expression by more than 30-fold within 24 h. This is in the same range, although with higher variability, as detected with fresh liver slices. In spite of considerable variability among the thawed slices, the induction factors are high enough for a sensitive detection of an induction at mRNA level. Additionally, immunostaining of respective CYP-forms was performed in sections of few samples, indicating CYP increase in viable cells of cryopreserved slices. PMID- 12093616 TI - Nerve growth factor protects against aluminum-mediated cell death. AB - In the present study, we examined the effect of two salts of aluminum (Al), aluminum maltolate (Almal) and aluminum chloride (AlCl(3)), on the cell viability of PC12 cells in the absence and presence of nerve growth factor (NGF). A 72-h exposure of PC12 cells to Almal (300 microM) resulted in a marked increase of lactic dehydrogenase (LDH) release from the cells and a decrease of 3-(4,5 dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) activity. These results indicate that Almal induces a decrease in the cell viability. Under the same conditions, Almal also caused DNA ladder formation and chromatin condensation. In contrast, AlCl(3) did not showed an increased LDH release and a decreased MTT activity in the concentration range of the salt tested (0.1-1 mM). The extent of LDH release and MTT activity decrease induced by Almal treatment closely depended on the amount of Almal incorporated into the cells. An increase in the fluorescence intensity of 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate, di(acetoxymethyl ester) (C-DCDHF-DA) which was loaded into the cell by Almal treatment and its prevention by pyrrolodine dithiocarbamate, a potent antioxidant, suggested that Almal-induced cell death partly proceeds via reactive oxygen species (ROS) production. NGF effectively inhibited the increase of LDH release and the decrease of MTT activity, as well as DNA fragmentation and chromatin condensation. However, NGF did not inhibit the increase of C-DCDHF-DA fluorescence in the cells induced by Almal treatment. From these results, it is suggested that ROS production associated with accumulation of Al is one possible important factor in the onset of Al neurotoxicity via apoptotic cell death and that NGF protects against cell degeneration associated with Al accumulation, but independently of ROS production. PMID- 12093617 TI - Effects of the porphyrinogenic compounds hexachlorobenzene and 3,5 diethoxycarbonyl-1,4-dihydrocollidine on polyamine metabolism. AB - The naturally occurring polyamines--putrescine, spermidine and spermine--are organic cations present in all living cells and essential for cell growth and differentiation. The aim of the present study was to extend the investigations on the effects of porphyrinogenic compounds on polyamine metabolism. This was achieved by studying putrescine, spermidine and spermine levels in a model of acute porphyria, i.e. 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC)-induced porphyria, and in a model of non-acute porphyria, i.e. hexachlorobenzene (HCB) induced porphyria. HCB administration to female Wistar rats for 7, 14, 21, 28 and 56 days did not alter polyamine levels in liver, even though rats presented clear signs of HCB-induced porphyria. In contrast to HCB, DDC treatment resulted in a remarkable increase in putrescine levels in the liver of female and male Sprague Dawley rats. This increase was due, at least in part, to ornithine decarboxylase (ODC) activation. DDC induction of putrescine levels did not show organ specificity, since it could also be seen in adrenal gland. Interestingly, the deregulation of polyamine biosynthesis occurred concomitantly with the deregulation of the heme biosynthetic pathway. In addition to porphyria, it is known that DDC intoxication affects several proteins of the hepatocyte cytoskeleton. It is suggested that DDC-induced increase in ODC activity and putrescine levels may be an early event contributing to alter the cytoskeleton. PMID- 12093618 TI - Presence of phenol UDP-glucuronosyltransferase in bovine alveolar macrophages and bronchial epithelial cells. AB - Pulmonary organ and cells are the primary target of atmospheric pollutants. Phenol UDP-glucuronosyltransferase (UGT) was found in bovine alveolar macrophage cells by immunohistochemical staining and also was observed in bronchial epithelial cells of the lung. A high level of activity of UGT, which is one of the phase II drug-metabolizing enzymes, toward 1-naphthol was observed in the microsomes of both cell types. By Western blotting analysis, a 54-kDa band was detected in alveolar macrophage cells and in bovine lung using polyclonal antibodies against a purified rat UGT, which catalyze the glucuronidation of various phenolic xenobiotics such as 1-naphthol and have the same molecular mass (54 kDa). Reverse transcriptase-polymerase chain reaction (RT-PCR) amplified the common cDNA region in UGT1A subfamily isoforms, indicating that UGT1A subfamily isoform was expressed in alveolar macrophages and in bronchial epithelial cells of the lung. These results suggest that phenol UGT act as a primary barrier against various phenolic chemicals in the lung. PMID- 12093620 TI - Lessons from chronic intermittent and sustained hypoxia at high altitudes. AB - Recurrent sleep apnea (RSA), mimicking chronic intermittent hypoxia (CIH), may trigger unique adaptations in oxygen sensing in the carotid body, and consequent cellular functions unlike the effects of sustained hypoxia (SH). As a mechanism, an augmented generation of reactive oxygen species (ROS) in CIH has been invoked at the exclusion of SH effects. The ROS might act at hypoxia inducible factors (HIF-1s), giving rise to various genes whose function is to restore the tissue P(O(2)) close to the original. In a spate, review articles on the CIH effects at sea level have appeared but little on high altitude (HA). Their views have been reexamined with the primary focus on the peripheral chemoreception. At HA, RSA is more common in the lowlanders because of a high ventilatory sensitivity to hypoxia (with the consequent effects) unlike the high altitude natives (HAN). Undoubtedly, the HIF-1s play a central role at HA, the mechanisms of which are unknown and explorable. PMID- 12093619 TI - Estradiol enhances and estriol inhibits the expression of CYP1A1 induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in a mouse ovarian cancer cell line. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous pollutant and promoter of carcinogenesis. This study investigated the interaction between TCDD and different estrogens in a cancer cell line (ID8) derived from mouse ovarian epithelium. TCDD-induced ethoxyresorufin-O-deethylase (EROD) activity and cytochrome P4501A1 (CYP1A1) expression in a dose- and time-dependent manner. Estrogen receptor (ER) alpha mRNAs were constitutively expressed, but ER beta and progesterone receptor (PR) mRNAs were not expressed. Induction of EROD by TCDD was completely inhibited by a alpha-naphthoflavone and phenanthroline, two aryl hydrocarbon receptor (AhR) antagonists. Progesterone and gonadotropins (FSH and LH) had no effect on the induction of EROD by TCDD. Congeners of 17beta-estradiol (E2) increased the induction of EROD activity by TCDD dose-dependently in the relative potency order: estrone (El)>E2> or = 4-hydroxyestradiol (4OHE2)> or = 2 hydroxyestradiol (2OHE2). In contrast, estriol (E3) decreased EROD activity induced by TCDD. E2 increased TCDD-induced CYP1A1 protein and mRNA whereas E3 decreased both the protein and mRNA. E2 did not alter luciferase activity induced by TCDD in cells transfected with a luciferase reporter containing dioxin response elements (DRE) or a CYP1A1 promoter. In contrast, E3 dose-dependently decreased the luciferase activity. A pure anti-estrogen (ICI 182780) inhibited the interaction between E2 and TCDD but did not block E3's effect on EROD activity. These results indicate that E2 may affect TCDD-induced CYP1A1 expression by a mechanism different from E3 in ID8 cells. It appears that the potentiation of E2 in the induction of CYP1A1 by TCDD occurs by a mechanism involving ER alpha since a specific ER antagonist blocked the potentiation. The inhibitory effect of E3 may be due to a rapid direct effect on EROD and a later suppression of CYP1A1 expression. PMID- 12093622 TI - Neurochemical perspectives on the control of breathing during sleep. AB - A specific depression of minute ventilation occurs during sleep in normal subjects. This sleep-related ventilatory depression is partially related to mechanical events and upper airway atonia but some data also indicate that it is likely to be centrally mediated. This paper reviews the anatomical and neurochemical connections between sleep/wake- and respiratory-related areas in an attempt to identify the potential implication of sleep-related neurochemicals (serotonin, catecholamines, GABA, acetylcholine) in the sleep-related hypoventilation. The review of available data suggests that the sleep-related ventilatory depression depends upon the enhanced GABAergic activity together with a loss of suprapontine influence depending on the cessation of activity of the reticular formation. During REM sleep, an additional inhibitory activity emerges from the pontine cholinergic neurons, which contributes to the breathing irregularities and the associated depression of minute ventilation and ventilatory response to chemical stimuli. This model may contribute to a better understanding of the neurochemical environment of respiratory neurons during sleep, which remains a question of importance regarding the numerous pathological states that are linked to specific perturbations of breathing control during sleep. PMID- 12093621 TI - Modulation of expiratory motor output evoked by chemical activation of pre Botzinger complex in vivo. AB - We have previously demonstrated that chemical stimulation of the pre-Botzinger complex (pre-BotC) in the anesthetized cat produces either phasic or tonic excitation of phrenic nerve discharge. This region is characterized by a mixture of inspiratory-modulated, expiratory-modulated, and phase-spanning (including pre inspiratory (pre-I)) neurons; however, its influence on expiratory motor output is unknown. We, therefore, examined the effects of chemical stimulation of the pre-BotC on expiratory motor output recorded from the caudal iliohypogastric (lumbar, L(2)) nerve. We found that unilateral microinjection of DL-homocysteic acid (DLH; 10 mM; 10-20 nl) into 16 sites in the pre-BotC enhanced lumbar nerve discharge, including changes in timing and patterning similar to those previously reported for phrenic motor output. Both increased peak amplitude and frequency of phasic lumbar bursts as well as tonic excitation of lumbar motor activity were observed. In some cases, evoked phasic lumbar nerve activity was synchronized in phase with phrenic nerve discharge. These findings demonstrate that chemical stimulation of the pre-BotC not only excites inspiratory motor activity but also excites expiratory motor output, suggesting a role for the pre-BotC in generation and modulation of inspiratory and expiratory rhythm and pattern. PMID- 12093624 TI - Diaphragm compound muscle action potential measured with magnetic stimulation and chest wall surface electrodes. AB - To seek a method to reliably measure phrenic nerve conduction time (PNCT) with magnetic stimulation we investigated two stimulus sites, placing the magnetic coil at the cricoid cartilage (high position) or close to the clavicle (low position). We also compared compound muscle action potential (CMAP) recorded from three different sites: in the sixth to eighth intercostal spaces in the anterior axillary line (Ant-a); in the 8th intercostal space close to the midclavicular line; and with one electrode at the lower sternum and the other at the costal margin. Fourteen normal subjects were studied. The PNCT measured by magnetic stimulation in the high position recorded from (Ant-a) was 7.6+/-0.6 on the left side and 8.4+/-0.7 on the right. The PNCT recorded from all three sites become much shorter when the magnetic coil was moved from the high to the low position. Our results show that PNCT can be accurately measured with magnetic stimulation when care is taken to avoid coactivation of the brachial plexus. PMID- 12093623 TI - CO(2)-induced c-Fos expression in brainstem preprotachykinin mRNA containing neurons. AB - Tachykinin peptides are found in brainstem regions involved in central chemoreception and they may play a modulatory role in ventilatory response to hypercapnia. We determined whether tachykinin peptide containing neurons are activated by CO(2) by combining in situ hybridization and immunohistochemistry (IHH). Experiments were performed in 21-day-old rats exposed to 12% CO(2) for 1 h. c-Fos expression was identified by IHH on free floating sections (40 microm) that were mounted and then hybridized with anti-sense 35S labeled ribonucleotide probe of the rat preprotachykinin A (PPT-A) gene. Sections were analyzed for expression of the PPT-A gene, c-Fos protein and colocalization of PPT-A gene with c-Fos protein. Within the chemosensory region of the nucleus tractus solitarius (nTS), 19% of c-Fos positive cells expressed PPT-A mRNA after hypercapnic loading. In medullary raphe nuclei, 64% of c-Fos positive cells expressed the PPT A gene after exposure to CO(2), while 21% of c-Fos labeled neurons in parapyramidal nuclei also expressed PPT-A mRNA. These results indicate that a subpopulation of CO(2) activated neurons within the nTS and in the parapyramidal and midline regions of the ventral aspect of the medulla oblongata express the PPT-A gene, suggesting that these are substance P- or neurokinin A-containing neurons. Furthermore, these peptides may play a role in modulation of respiratory and cardiovascular responses to changes in CO(2)/H(+) content of the extracellular fluid. PMID- 12093625 TI - Respiratory response to mechanical stimulation of the gallbladder. AB - Since stimuli from abdominal or pelvic viscera can affect respiratory muscle function, we hypothesized that mechanical stimulation of the gallbladder would result in inhibition of motor activity to the diaphragm and to upper airway muscles. We studied 12 decerebrate, vagotomized, paralyzed, artificially ventilated cats and recorded hypoglossal (HG) and phrenic (PHR) nerve activities while applying 600-1000 g of traction on the gallbladder during four respiratory cycles. Traction resulted in an initial reduction of PHR activity to 87.6+/-15.0% (mean+/-S.D.% of its baseline value), a reduction of HG activity to 74.2+/-27.5% and a lengthening of expiratory time to 178.8+/-81.0%. Subsequently, PHR activity and expiratory time returned toward control values, while HG remained diminished, at 66.4+/-19.1%. Our results show that mechanical stimulation of the gallbladder results in a respiratory inhibition with a disproportionate reduction in HG activity relative to PHR discharge. We speculate that gallbladder stimulation by contractions or surgery may compromise breathing by inhibition of phrenic discharge and upper airway obstruction. PMID- 12093626 TI - Effects of abdominal distension on breathing pattern and respiratory mechanics in rabbits. AB - The effects of acute abdominal distension (AD) on the electromechanical efficiency (Eff) of the inspiratory muscles were investigated in anesthetized rabbits by recording the electrical activity (A), pressure (P) exerted by the diaphragm (di) and parasternal intercostal muscles (ic), and lung volume changes when an abdominal balloon was inflated to various degrees. Eff,ic increased with increasing AD both in supine and upright postures. In upright rabbits Eff,di increased for intermediate but decreased at higher levels of AD, whilst it decreased at all levels of AD in supine rabbits. Tidal volume (VT) response followed that of Eff,di. Tonic Aic and Adi and inspiratory prolongation were elicited by AD. The effects of these neural mechanisms, acting to limit end expiratory lung volume and VT changes, were however small since vagotomy prevented tonic Adi and inspiratory prolongation and reduced tonic Aic, but changed lung volume responses to AD only little. Hence, reduced respiratory system compliance and changes in inspiratory muscle electromechanical efficiency dominate lung volume responses to acute AD. PMID- 12093627 TI - Effect of chest wall vibration on dyspnea during exercise in chronic obstructive pulmonary disease. AB - To elucidate the effect of in-phase chest wall vibration (IPV) during exercise, 17 COPD male patients performed two constant-load exercise tests on a cycle ergometer with and without IPV. The Borg dyspnea score significantly decreased from IPV (-) to IPV (+) (from 13.6+/-2.9 to 12.5+/-2.9, P<0.01). IPV elicited a significant increase in V(O(2)) (P<0.005) and significant decreases in both VE/V(O(2)) (P<0.05) and respiratory frequency (P<0.05), but it did not elicit any changes in VE. The change in Borg score between IPV (+) and IPV (-) showed a significant positive correlation with % predicted V(O(2),max) (r=0.71) and FEV(1)/FVC (r=0.69). Patients in the responsive group (n=11) showed significantly lower FEV(1) (P<0.05) and higher DeltaN(2)/L (P<0.01) than patients in the non responsive group (n=6). We conclude that IPV reduces dyspnea and improves respiratory efficiency during aerobic exercise in severe COPD. PMID- 12093628 TI - Geometric determinants of airway resistance in two isomorphic rodent species. AB - The flow resistance of the pulmonary airway tree (Raw) is disproportionately larger in large animals than in smaller ones. This is thought to be due to the fact that smaller animals have relatively wider central airways than larger animals. However, Raw is not determined solely by the diameter of the main bronchi or trachea. It depends on the dimensions of all the individual airways and how they are connected. We investigated how the degree of asymmetry of the airway tree, the number of airway orders, and dimensions of the individual airways combine to determine Raw. We performed this investigation using computer models of the airway trees of two different sized isomorphic rodents that differ in weight by more than two orders of magnitude-the harvest mouse and the giant pouched rat. We calculated airway resistance in these models under various conditions, and confirmed that the smaller species has a lower Raw relative to body size than the larger species. We also showed that these differences are due to a combination of differences both in relative airway diameters and in the degree of asymmetry of the airway trees. PMID- 12093629 TI - An invariant number of the respiratory system of mammals: newborn and adult. AB - From four empirical allometric equations concerning the dynamics of the respiratory functions of mammals, it has been possible to obtain an invariant and dimensionless number after applying Buckingham's pi-theorem. In the present study, this invariant number (IN(R)), whose origin was interspecies comparisons in mammals of different sizes, was assayed with the aim to compare in a quantitative manner the possible difference between newborn and adult mammals. The results were compared with the predicted values from two theories of biological similarity, one mass-dependent, valid for newborns, and the other, weight-dependent, valid for adult mammals. Finally, we utilized Stahl's residual mass exponents (RME) to test the validity of the empirical and theoretical approaches. PMID- 12093631 TI - Cataract extraction and IOL implantation. PMID- 12093634 TI - Optic disc morphology after AAION. PMID- 12093632 TI - Optic disc morphology after AAION. PMID- 12093636 TI - Research: the track record justifies expanded support. PMID- 12093637 TI - Clinical recommendations for repair of isolated orbital floor fractures: an evidence-based analysis. AB - PURPOSE: To assess the quality of information in the literature and suggest guidelines for repair of isolated orbital floor fractures. CLINICAL RELEVANCE: Orbital floor fractures are a common result of orbital injury. Enophthalmos, diplopia resulting from extraocular muscle dysfunction, and infraorbital nerve hypesthesia may occur. The indications and timing for fracture repair are still controversial. LITERATURE REVIEWED: A MEDLINE literature review was performed using PubMed. Articles published from 1983 to the present were retrieved using the key words, "orbital floor fracture, orbital trap-door fracture, and orbital blow-out fracture." Suggested indications and timing for repair of isolated orbital floor fractures were extracted from selected articles. Each recommendation was rated according to its importance in the care process and strength of evidence supporting the given recommendation. RESULTS: No prospective randomized clinical trials on the treatment of orbital floor fractures have been performed. Despite this, most recommendations were rated as most important to patient care (A) and had strong support for treatment (level I). CONCLUSIONS: The timing and treatment indications for orbital floor fractures are evolving. Nonresolving oculocardiac reflex, the "white-eyed" blowout fracture, and early enophthalmos or hypoglobus are indications for immediate surgical repair. Surgery within 2 weeks is recommended in cases of symptomatic diplopia with positive forced ductions and evidence of orbital soft tissue entrapment on computed tomography examination or large orbital floor fractures that may cause latent enophthalmos or hypo-ophthalmos. PMID- 12093640 TI - Routine histopathologic evaluation of the lacrimal sac during dacryocystorhinostomy: how useful is it? AB - OBJECTIVE: To determine the usefulness of "routine" lacrimal sac biopsy to evaluate unsuspected neoplasm or systemic disease in patients with acquired nasolacrimal duct obstruction. DESIGN: Noncomparative, interventional case series with histopathologic correlation. PARTICIPANTS: Review of charts revealed histopathologic reports on 302 specimens from 258 patients who had undergone routine dacryocystorhinostomy for clinical acquired nasolacrimal duct obstruction. INTERVENTION: Dacryocystorhinostomy (DCR) with histopathologic evaluation of lacrimal sac. MAIN OUTCOME MEASURES: The following variables were extracted from the medical records: age, gender, presenting symptoms, history of predisposing conditions, lacrimal system abnormalities noted before surgery, dye disappearance test, dacryocystogram or dacryoscintillogram, appearance of the lacrimal sac during surgery, and histopathologic report of the lacrimal sac specimen. RESULTS: Findings of histopathologic evaluation of 302 lacrimal sac specimens from 170 females and 88 males who had undergone routine DCR were reviewed. Evidence of systemic disease or neoplasia involving the lacrimal sac was present in 10 specimens. All 10 positive specimens had a grossly abnormal appearing sac at the time of surgery (n = 8), a known preexisting history of systemic diseases (n = 6), or both. The remaining 292 specimens showed chronic inflammation, fibrosis, or normal mucosa. CONCLUSIONS: Only patients with known preexisting systemic disease or a grossly abnormal lacrimal sac had "positive" histopathologic findings. We recommend that lacrimal sac biopsy in patients undergoing DCR should be performed only in those with a positive history for systemic disease or an abnormal-appearing lacrimal sac during surgery. PMID- 12093642 TI - Graded orbital decompression based on severity of proptosis. AB - OBJECTIVE: To study the results of orbital decompression based on the severity of preoperative proptosis. DESIGN: A retrospective noncomparative interventional case series. PARTICIPANTS: Thirty-nine orbits in 23 patients with thyroid-related orbitopathy at a university-based referral center. INTERVENTION: Graded orbital decompression was performed in all patients based on the severity of preoperative exophthalmometry. MAIN OUTCOME MEASURES: Exophthalmometry, visual acuity, margin to-reflex distance, prism cover testing, and intraocular pressure. RESULTS: Mean proptosis reduction in all orbits was 6.4 +/- 2.7 mm (P < 0.01). In group 1 (preoperative exophthalmometry <22 mm), proptosis decreased with a mean of 4.8 +/ 1.3 mm (P < 0.01); mean proptosis reduction was 6.0 +/- 2.3 mm (P < 0.01) and 8.9 +/- 3.4 mm (P < 0.01) in group 2 (exophthalmometry between 22-25 mm) and group 3 (exophthalmometry >25 mm), respectively. In four of five eyes with compressive optic neuropathy there was an improvement of best-corrected visual acuity of 2 lines or more. Margin-to-reflex distance of the upper and lower lids and intraocular pressure were reduced in all groups. New-onset diplopia developed in two patients (8.7%); 13 of 15 patients (86.7%) who had diplopia preoperatively had persistent diplopia postoperatively. Two patients (13.3%) had relief of diplopia postoperatively. CONCLUSIONS: Graded orbital decompression based on the severity of preoperative exophthalmometry is useful to determine the type and amount of orbital surgery to be performed. PMID- 12093643 TI - The association of atherosclerosis, vascular risk factors, and retinopathy in adults with diabetes : the atherosclerosis risk in communities study. AB - PURPOSE: The aim of this report is to describe the prevalence of retinopathy and its associations with atherosclerosis and vascular risk factors in people with diabetes. DESIGN: Cross-sectional study. PARTICIPANTS: Persons with diabetes, having gradable fundus photographs, from a biracial population-based cohort of adults (ages 51-72 years), and living in four United States communities (Forsyth County, North Carolina; the city of Jackson, Mississippi; suburbs of Minneapolis, Minnesota; and Washington County, Maryland) were studied from 1993 to 1995. METHODS: Lesions typical of diabetic retinopathy were detected by grading a 45 degrees color fundus photograph of one eye of each participant, using a modification of the Airlie House classification system. MAIN OUTCOME MEASURE: Severity of diabetic retinopathy (none, minimal nonproliferative, moderate nonproliferative, severe nonproliferative, and proliferative) and macular edema. RESULTS: Retinopathy was detected in 328/1600 (20.5%) of those with diabetes; 114/1724 (6.6%) had hard exudate, 28/1600 (1.8%) had proliferative diabetic retinopathy, and 27/1662 (1.6%) had macular edema. The prevalence of diabetic retinopathy was higher in blacks (27.7%) compared with whites (16.7%). Controlling for duration of diabetes, serum glucose, systolic blood pressure, and type of diabetes medications taken, severity of retinopathy was associated with carotid artery intima-media wall thickness (odds ratio [OR]/0.1-mm thickness 1.09; 95% confidence interval [CI], 1.01, 1.17; P = 0.01), serum albumin (OR/0.1 g/dl 0.94; 95% CI, 0.88, 0.99; P = 0.02), but not race (OR blacks versus whites,1.24; 95% CI, 0.88, 1.75; P = 0.21). Severity of diabetic retinopathy was not associated with coronary artery disease or stroke history or any of the plasma lipids studied. Controlling for age, gender, duration of diabetes, serum glucose, and type of diabetes medications taken, the presence of retinal hard exudates was associated with plasma low-density lipoprotein cholesterol (OR/10 mg/dl 1.18; 95% CI, 1.09, 1.29; P < 0.001), and plasma Lp(a) (OR/10 mg/dl 1.02; 95% CI, 1.00, 1.05; P = 0.04) but not race or blood pressure. CONCLUSIONS: These data suggest that plasma lipids are associated with the presence of hard exudate and that carotid artery intima-media wall thickness is associated with retinopathy, but other manifestations of atherosclerosis and most of its risk factors are not associated with severity of diabetic retinopathy. PMID- 12093645 TI - Localized retinal dysfunction in central serous chorioretinopathy as measured using the multifocal electroretinogram. AB - PURPOSE: To determine the extent of electrophysiologic dysfunction in patients with central serous chorioretinopathy (CSC). DESIGN: Prospective observational case series. PARTICIPANTS: Six patients with unilateral CSC (mean age, 40 years) were recruited into the study. METHODS: Six patients with CSC underwent multifocal electroretinogram (mfERG) testing on both their clinically affected and opposite uninvolved eyes using the VERIS System, with a stimulus array of 103 scaled hexagons. The first positive peak responses were analyzed within six concentric ring annuli centered on the fovea. Amplitudes and implicit times were compared with those of an age-similar control group. MAIN OUTCOME MEASURES: Local electroretinographic response amplitudes and implicit times within the central 40 degrees with the mfERG. RESULTS: All the clinically uninvolved eyes showed mfERG amplitudes and implicit times within the normal range throughout the central 40 degrees of the retina. All six eyes with CSC showed reduced amplitudes and/or delayed implicit times that were limited to the regions of the macula in which clinical changes associated with CSC were apparent. CONCLUSIONS: We observed electroretinographic changes only in the clinically affected eyes, and these were limited to regions with ophthalmoscopically apparent fundus changes. Our findings do not support the conclusion that functional impairment, as measured by the mfERG, in eyes with CSC extends beyond clinically observed fundus changes. We did not observe abnormal mfERG responses in the clinically normal eyes of such patients. PMID- 12093644 TI - Early age-related maculopathy and self-reported visual difficulty in daily life. AB - PURPOSE: To determine whether early age-related maculopathy (ARM) is associated with visual difficulty in daily activities beyond the difficulty that would be expected based on normal retinal aging; to determine whether scotopic sensitivity and visual acuity are associated with visual difficulties in these older adults. STUDY DESIGN: Comparative, cross-sectional questionnaire study. SUBJECTS: Ninety two older adults with early ARM in at least one eye as defined by one or more large (>63 microm) drusen and/or focal hyperpigmentation but no choroidal neovascularization or geographic atrophy, acuity of 20/60 or better, and a reference group of 55 older adults in the same age range without these fundus features and acuity of 20/35 or better in each eye. METHOD: Tests of visual acuity and scotopic sensitivity and a general health questionnaire were carried out. The Activities of Daily Vision Scale (ADVS) was administered to assess self reported visual difficulties in everyday tasks and expressed on a scale of 0 (extreme difficulty) to 100 (no difficulty). Fundus photographs were taken and graded to characterize the presence and severity of ARM to determine eligibility. RESULTS: For purposes of analysis, the early ARM group was divided into those whose fellow eye (FE) was 20/60 or better and those whose FE was worse than 20/60. ADVS subscale scores were substantially lower in the early ARM group with FE worse than 20/60 (medians, 58-83) compared with the normal retinal health group (medians, 97-100). Even for those with early ARM with FE 20/60 or better, four of five subscale scores were lower (medians, 81-97), albeit slightly in some cases, than those of the reference group. For both ARM subgroups, the night driving subscale had the lowest scores of all subscales. Persons with early ARM with FE 20/60 or better were more likely to report difficulty on the night driving (odds ratio [OR], 4.3; 95% confidence interval [CI], 1.6-11.4), near vision (OR, 5.0; 95% CI, 1.9-12.9), and glare disability (OR, 2.7; 95% CI, 1.1 6.3) subscales compared with those in normal retinal health, adjusting for age, gender, medical comorbidities, and lens density. For early ARM patients with FE worse than 20/60, there was widespread reporting of difficulty on all subscales (ORs ranging from 4.7-52.9). Poor scotopic sensitivity was highly associated with difficulty on the night driving subscale (OR, 6.6; 95% CI, 1.2-35.5) but not with any other subscale. Acuity worse than 20/25 in both eyes was significantly associated with difficulty on all ADVS subscales; when this acuity impairment was present in one eye only, associations were still significantly present on some subscales, although they were weaker. CONCLUSIONS: Persons in the early phases of ARM, even when their fellow eye has relatively good acuity, are more likely to experience difficulty in night driving, near vision tasks, and glare disability compared with those in good retinal health. Scotopic dysfunction, a functional marker of early ARM, is linked to reported night driving problems. Even when acuity impairment occurs in one eye only, patients report difficulties with day driving and near and far vision tasks. PMID- 12093646 TI - Posterior vitreous findings in cases of spontaneous retinal reattachment. AB - PURPOSE: To describe the posterior vitreous findings in two patients with retinal detachment who experienced spontaneous retinal reattachment. DESIGN: Two observational case reports. TESTING: Biomicroscopic and high-resolution echographic evaluation of the vitreoretinal relationships. MAIN OUTCOME MEASURES: Retinal reattachment and echographic vitreoretinal relationships. RESULTS: Two patients, one with a rhegmatogenous retinal detachment and one with a tractional retinal detachment, were noted to have a partial posterior vitreous detachment with vitreoretinal adherence at the time of presentation. Spontaneous retinal reattachment occurred in both cases, with echography showing complete vitreous separation from the retina. CONCLUSIONS: Complete posterior vitreous detachment may release tractional components in retinal detachment and contribute to spontaneous retinal reattachment. PMID- 12093647 TI - Benefits and complications of photodynamic therapy of papillary capillary hemangiomas. AB - OBJECTIVE: To evaluate the potential benefit and risks of photodynamic therapy (PDT) in the treatment of papillary capillary hemangioma. DESIGN: Prospective, noncomparative, interventional case series. PARTICIPANTS: Five patients with solitary capillary hemangioma on the temporal portion of the optic nerve presenting with exudative decompensation and decrease in visual acuity (VA). METHODS: All eyes received a standardized PDT treatment with 6 mg/kg body surface area verteporfin and application of 100 J/cm(2) light at 692 nm. One to three PDT courses were performed until resolution of exudation was achieved. A continuous follow-up was provided with documentation 1 week before and at 4 to 6 weeks, 3 months, and 12 months after the last treatment application. MAIN OUTCOME MEASURES: Functional parameters included best-refracted VA (Early Treatment Diabetic Retinopathy Study), and central scanning laser ophthalmoscope (SLO) scotometry and peripheral (automated perimetry) visual fields; anatomic parameters were presence of retinal edema or serous detachment (ophthalmoscopy) and tumor size (ultrasonography). RESULTS: Pretreatment VA levels ranged from 20/40 to 20/800; posttreatment levels ranged from 20/64 to 20/2000. Tumor regression with resolution of macular exudate and serous retinal detachment was obtained in all eyes. A decline in VA of 1, 3, and 10 lines, respectively, was documented in three patients. Complications included transient decompensation of vascular permeability, occlusion of retinal vessels, and ischemia of the optic nerve. CONCLUSIONS: PDT is successful in reducing tumor size and exudative activity. Vaso-occlusive effects at the level of the retina and optic nerve compromise the functional benefit. Parameters proven safe in choroidal neovascularization may be inappropriate in retinal capillary lesions of the optic nerve. PMID- 12093648 TI - Thrombocytosis in patients with biopsy-proven giant cell arteritis. AB - OBJECTIVE: To compare the platelet counts, complete blood counts, and Westergren sedimentation rates (WESR) of patients with a biopsy positive for giant cell arteritis (GCA) with those of patients with negative biopsies. DESIGN: Retrospective, case-control series. METHOD: The medical records of 91 consecutive patients who underwent temporal artery biopsy for possible GCA, over an 8-year period, were reviewed. MAIN OUTCOME MEASURES: Values obtained for the complete blood counts, including platelet counts, WESR, and biopsy results of 91 consecutive patients undergoing temporal artery biopsy were analyzed. RESULTS: The mean platelet count of 47 patients with positive temporal artery biopsies (433 x 10(3)/microl) was significantly higher than that of 44 patients with negative temporal artery biopsies (277 x 10(3)/microl), P < 0.0001. There was no statistically significant difference in the mean WESR between the biopsy-positive (82 mm/hour) and biopsy-negative (70 mm/hour) groups, P = 0.12. The sensitivity of an elevated WESR for biopsy-positive patients was greater (79%; 95% confidence interval [CI], 64%-89%) than that of an elevated platelet count (57%; 95% CI, 42% 72%). However, the specificity (91% [95% CI, 78%-97%] versus 27% [95% CI, 15% 43%]), positive predictive value (87% [95% CI, 70%-96%] versus 54% [95% CI, 41% 66%]), and negative predictive value (67% [95% CI, 53%-78%] versus 55% [95% CI, 32%-76%]) favored an elevated platelet count compared with WESR, or to the combination of platelets and WESR, as a better test for diagnosing GCA in the 91 patients studied. The area under the receiver operating characteristic function for platelets (0.72) was greater than that for WESR (0.59) or the combination of platelets and WESR (0.65). CONCLUSIONS: In patients suspected of having GCA, an elevated platelet count greater than 400 x 10(3)/microl is a useful marker of a positive temporal artery biopsy. PMID- 12093649 TI - Choroidal vascular occlusion in a child with a connective tissue disease and complement C4 deficiency. AB - OBJECTIVE: To report the histopathologic findings in the eyes of a patient with a connective tissue disease and complement deficiency. DESIGN: Human postmortem ocular histopathologic study. INTERVENTION: A 15-year-old female died from complications of a connective tissue disease of uncertain etiology, particularly acute respiratory distress syndrome. Abnormalities seen in the eyes at autopsy were consistent with complement activation, granulocyte aggregation, and leukocyte embolization. MAIN OUTCOME MEASURES: Both eyes were examined by light microscopy. RESULTS: Some choroidal vessels were occluded by platelet-fibrin thrombi and occasionally by aggregates of granulocytes and fibrin. Serous retinal detachment involving the macula and peripheral retina was present in both eyes. CONCLUSIONS: This is a report of the ocular histopathologic findings in a patient with connective tissue disease and complement C4 deficiency. The light microscopy findings were consistent with complement activation with granulocyte aggregation and leukocyte embolization and may represent another mechanism to explain the clinical findings in patients with connective tissue disease, particularly systemic lupus erythematosus. PMID- 12093650 TI - Long-term outcomes of keratolimbal allograft for the treatment of severe ocular surface disorders. AB - PURPOSE: To investigate the long-term outcome of keratolimbal allograft (KLAL) for the treatment of severe ocular surface disorders. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: Twenty patients (23 eyes) with severe ocular surface disorders. INTERVENTION: Thirty-three KLAL procedures were performed. Ten patients (10 eyes) underwent KLAL in combination with other surgical procedures. Oral or topical cyclosporine or both were used after surgery in 15 patients to prevent allograft rejection. MAIN OUTCOME MEASURES: Reconstruction of the ocular surface with restoration of phenotypic corneal epithelium, reduction of corneal vascularization and conjunctivalization, decreased pain, and visual improvement. RESULTS: The mean follow-up was 60 months (range, 15-96 months). Eight eyes (24.2%) never reepithelialized and were considered primary failures. The remaining 25 grafts initially restored a phenotypic corneal epithelium, but at last follow-up only 7 (21.2%) were stable. Graft survival rate was 54.4% at 1 year, 33.3% at 2 years, and 27.3% at 3 years. Visual acuity improved or was unchanged in 19 eyes (82.6%) and decreased in 4 eyes (17.4%). Seventeen corneal transplantations (3 lamellar keratoplasties and 14 penetrating keratoplasties) were performed either in combination with or after a KLAL. All three lamellar keratoplasties were successful, whereas 13 of the 14 penetrating keratoplasties failed. Cyclosporine was used initially in high-risk recipients and later in all recipients. Allograft rejection episodes occurred in 13 KLAL procedures of 11 eyes (39.4%) and were more common in patients treated with cyclosporine compared with the untreated group (87.5% vs. 22.2%). Graft survival was longer in the cyclosporine-treated group compared with the untreated group. CONCLUSIONS: Keratolimbal allograft is useful in ocular surface reconstruction and restores phenotypic corneal epithelium. Graft survival rate, however, decreases dramatically over a 2-year period. Long-term use of cyclosporine appears to prolong graft survival but does not prevent acute allograft rejections. PMID- 12093651 TI - Transplantation of human limbal epithelium cultivated on amniotic membrane for the treatment of severe ocular surface disorders. AB - PURPOSE: To study the short-term clinical results of transplanting of cultivated corneal/limbal epithelial cells on human amniotic membrane (AM) for limbal deficiency. DESIGN: Noncomparative, retrospective interventional case series. PARTICIPANTS: Thirteen eyes of 13 patients with severe limbal deficiency (Stevens Johnson syndrome in eight eyes, ocular cicatricial pemphigoid in three eyes, and chemical burns in two eyes) were treated at the department of Ophthalmology, Tokyo Dental College, Japan. INTERVENTION: Cultivated allo-limbal epithelium was transplanted onto the ocular surface of patients with severe limbal deficiency. MAIN OUTCOME MEASURES: Ocular surface reconstruction with corneal epithelialization, changes in visual acuity, and postoperative complications were studied. Histologic examinations were also performed on cultivated epithelium. RESULTS: Cultivated corneal epithelium on AM formed two to three layers with the formation of basement membrane-like structures. After the surgery, the epithelium regenerated and covered the ocular surface in eight eyes (61.5%). However, three of the eight eyes developed partial conjunctival invasion, and two eyes later developed epithelial defects. At last examination, corneal epithelialization was achieved in six eyes (46.2%). Five eyes had conjunctivalization, one eye had dermal epithelialization, and one eye was not epithelialized. Complications were corneal perforation in four eyes and infectious keratitis in two eyes. CONCLUSIONS: This study demonstrates that the success rate for transplanting cultivated allo-limbal epithelium on the AM is not different from the conventional limbal and AM transplantation for the treatment of severe limbal stem cell dysfunction. PMID- 12093652 TI - Spontaneous wound dehiscence after removal of single continuous penetrating keratoplasty suture. AB - PURPOSE: To determine the incidence and complications of spontaneous wound dehiscence after removal of a single continuous penetrating keratoplasty (PK) suture. DESIGN: Retrospective consecutive, noncomparative interventional case series. METHODS: Retrospective review of 324 consecutive continuous suture PKs performed between 1992 and 1999. RESULTS: Sixty-nine (21.3%) of 324 PKs reviewed had the continuous suture removed. The average interval for suture removal after PK was 24.5 +/- 15 months (range, 2.8-63.3 months). Five of the 69 eyes (7.2%) developed spontaneous wound dehiscence without direct eye trauma. In the five eyes that developed wound dehiscence, the continuous suture was removed at 24.6 +/- 10.3 months (range, 14-42 months). Dehiscence occurred at 11.6 +/- 6.5 (range, 3-18) days after suture removal. Significant history associated with wound dehiscence included coughing, yawning, falling without trauma to the eye, and spontaneous wound separation. The reasons for suture removal were astigmatism in four of five (80%) patients and a broken suture in one of the five patients. In four of five (80%) patients, the location of wound dehiscence correlated with the steep axis of corneal keratometry before suture removal. Surgical intervention preserved the presuture removal best-corrected visual acuity in four of the five eyes. No eyes with an intact suture spontaneously dehisced. CONCLUSIONS: The rate of spontaneous wound dehiscence after removal of a continuous suture in our series was 7.2%. All spontaneous dehiscences occurred within 2 weeks after suture removal. Older patients, who had PK for corneal edema with postoperative astigmatism and have been using corticosteroids drops for prolonged periods of time, are at higher risk of wound dehiscence. Patients should be monitored closely during the first 2 weeks after removal of a continuous suture for signs of wound separation, especially when suture removal is performed for astigmatism. Patients should be cautioned about the risk and symptoms of wound dehiscence before suture removal to facilitate early recognition and intervention for preservation of best visual potential. PMID- 12093654 TI - Standardized color-coded scales for anterior and posterior elevation maps of scanning slit corneal topography. AB - PURPOSE: To find the most appropriate color-coded scales for the anterior and posterior elevation maps of scanning slit topography in the screening of abnormal corneas such as keratoconus. DESIGN: Retrospective case-control study. PARTICIPANTS: Eighty eyes of 40 normal subjects and 175 eyes of 95 patients with keratoconus. INTERVENTION: Anterior and posterior corneal elevations were assessed using Orbscan 2. Best-fit sphere maps were drawn with several color coded scales: 2-, 5-, 10-, and 20-microm height per each color interval. MAIN OUTCOME MEASURES: The maps were judged to be abnormal when more than three colors (discriminant number) were found within the central 3-mm area. For each color coded scale, sensitivity, specificity, positive predictive value, negative predictive value, and sensitivity + specificity were calculated. After determining the most appropriate color-coded scales for the anterior and posterior elevation maps, validity of the discriminant number was assessed. By varying the discriminant number from two to eight, receiver operator characteristic (ROC) curves were created using the sensitivity and specificity for each threshold number. RESULTS: The highest sensitivity + specificity values and highly balanced predictive values were obtained with the 10- and 20-microm scales for the anterior and posterior elevation maps, respectively. The ROC curve analyses showed that the best discriminant color number is three, indicating that maps with four or more colors within the central 3-mm area are judged abnormal in screening. CONCLUSIONS: The 10- and 20-microm interval color scales are most appropriate for the anterior and posterior elevation maps of the scanning slit topography, respectively. PMID- 12093656 TI - Use of augmented rectus muscle transposition surgery for complex strabismus. AB - PURPOSE: Full tendon rectus muscle transposition surgery augmented with posterior fixation sutures has been shown to be effective in the treatment of abducens palsy and Duane's syndrome. The purpose of this study is to summarize our experience with the use of this procedure and a three fourths partial tendon transposition modification of this procedure for a variety of complex vertical and horizontal paralytic eye movement disorders. DESIGN: Retrospective noncomparative interventional consecutive case series. PARTICIPANTS: Eighteen patients with paralytic strabismus. INTERVENTION: Full tendon rectus muscle transposition procedure augmented with posterior fixation sutures (13 patients) or a three fourths partial tendon transposition augmented with posterior fixation sutures (5 patients). MAIN OUTCOME MEASURES: Resolution of diplopia in the primary position and improved ocular alignment. RESULTS: Comparing preoperative to postoperative status, presence of primary position diplopia decreased from 64% to 14%, anomalous head posture decreased from 56% to 6%, and primary position alignment improved in all patients. CONCLUSIONS: Full tendon rectus muscle transposition surgery augmented with posterior fixation sutures and the vessel sparing three fourths partial tendon transposition modification of this technique are effective for the treatment of a variety of complex vertical and horizontal paralytic ocular motility disorders. PMID- 12093655 TI - Risk factors for incident nuclear opacities. AB - PURPOSE: To evaluate risk factors for the 4-year incidence of nuclear opacities. DESIGN: Population-based cohort study (85% participation at 4-year follow-up). PARTICIPANTS: Two thousand six hundred nine black participants of the Barbados Eye Studies, without any nuclear opacities at baseline. METHODS: Participants completed a standardized protocol at baseline and follow-up, including ophthalmic and other measurements, an interview, slit-lamp lens grading, fundus photography, and an ophthalmologic examination. Factors associated with the incidence of nuclear opacities (Lens Opacities Classification System II N > or = 2) were evaluated by logistic regression. MAIN OUTCOME MEASURE: Relative risks (RR) with 95% confidence intervals (95% CI). RESULTS: The 4-year incidence of nuclear opacities was 9.2% (241 of 2609) and increased greatly with age. Women were at significantly greater risk (RR = 1.8), as were persons with darker iris color (RR = 4.9), myopia (RR = 2.8), history of diabetes (RR = 1.6), leaner body mass (RR = 0.95 for each unit increase in body mass index [kg/m(2)]), and intraocular pressure (IOP)-lowering treatment (RR = 2.7), mainly with topical beta-blockers. Treated participants had a threefold RR of nuclear opacities (RR = 3.2; 95% CI, 1.6, 6.5) compared with those untreated and with IOP < or =21 mmHg. Among participants with IOP >21 mmHg, those receiving treatment (n = 33) had a fivefold RR (RR = 5.0; 95% CI, 1.7, 15.1) versus those who were untreated. The RR was similar for treated persons with and without open-angle glaucoma (RR = 3.1; 95% CI, 1.3, 7.4 and RR = 2.8; 95% CI, 0.9, 8.6 respectively) but was lower in persons with newly detected (and thus untreated) glaucoma at baseline (RR = 1.2; 95% CI, 0.6, 2.6) compared with those without open-angle glaucoma or treatment. CONCLUSIONS: The 4-year risk of nuclear opacities increased with age, female gender, darker iris color, myopia, diabetes, and leaner body mass, indicating similarities with other populations. The use of topical IOP-lowering medications tripled the RR of nuclear opacities in this study, an association that requires verification from clinical trials. PMID- 12093657 TI - Vertical misalignment in unilateral sixth nerve palsy. AB - OBJECTIVE: To detect and determine the magnitude of vertical deviation in patients with unilateral sixth nerve palsy. DESIGN: Prospective consecutive comparative case series. PARTICIPANTS: Twenty patients with unilateral peripheral sixth nerve palsy, 7 patients with central palsy caused by brainstem lesions, and 10 normal subjects. METHODS: Subjects were tested by the prism and cover test, Maddox rod and prism test, and magnetic search coil recordings in nine diagnostic eye positions. They were also tested during static lateral head tilt by the prism and cover, and Maddox rod and prism tests. MAIN OUTCOME MEASURES: The magnitudes of horizontal and vertical deviations. RESULTS: All patients had an abduction deficit and incomitant esodeviation that increased in the field of action of the paretic muscle, indicating sixth nerve palsy. Mean vertical deviations, for all positions of gaze in peripheral palsy were 0.3 +/- 0.8 prism diopters (PD) by prism and cover test, 1.3 +/- 1.6 PD by Maddox rod and prism test, and 2.0 +/- 1.4 PD by coil recordings. Mean vertical deviations in normal subjects were 0.0 +/- 0.0 PD by prism and cover test, 1.0 +/- 0.9 PD by Maddox rod and prism test, and 1.9 +/- 2.1 PD by coil recordings. Therefore, peripheral palsy did not cause abnormal vertical deviation. In central palsy, for all positions together mean vertical deviations were 0.9 +/- 1.3 PD by prism and cover test, 1.4 +/- 1.6 PD by Maddox rod and prism test, and 2.5 +/- 1.6 PD by coil recordings; they were not different from normal values. During static head roll, patients with peripheral palsy had a right hyperdeviation on right head tilt and a left hyperdeviation on left head tilt, regardless of the side of the palsy. In contrast, in central palsy, head tilt caused vertical strabismus that remained on the same side on head tilt to either side. CONCLUSIONS: Small vertical deviations in sixth nerve palsy are consistent with normal hyperphorias that become manifest in the presence of esotropia. In peripheral sixth nerve palsy, static head roll to either side induces hyperdeviation in the eye on the side of the head tilt. Hyperdeviation of the same eye induced by head tilt to either direction implicates a brainstem lesion as the cause of paretic abduction. Quantitative study of sixth nerve palsy demonstrates that if a vertical deviation falls within the normal range of hyperphoria, multiple cranial nerve palsy or skew deviation may not be responsible. Conversely, vertical deviation > 5 PD indicates skew deviation or peripheral nerve palsy in addition to abduction palsy. PMID- 12093658 TI - Clinical features and natural history of the acute idiopathic enlarged blind spot syndrome. AB - OBJECTIVE: To study the clinical pattern and natural history of patients with the symptom of an enlarged blind spot. DESIGN: A retrospective case series. Twenty one patients were collected from a neuro-ophthalmologic practice during the period January 14, 1983, to July 1, 1996, and four consecutive patients were added from three vitreoretinal practices during the period April 14, 1986, to June 7, 1999. PARTICIPANTS: Twenty-six eyes of 25 patients were studied at onset and at repeat visits from 1 year and 7 months to 15 years and 6 months later. METHODS: The first visit was composed of a complete neuro-ophthalmologic examination with fundus photos and fluorescein angiography in 12 of the 26 eyes. Follow-up examination consisted of an interval history, ophthalmologic examination, visual fields, fundus photographs, fluorescein angiograms in eight eyes, indocyanine green angiograms in seven eyes, and multifocal electroretinograms (mfERG) in both eyes of seven patients. MAIN OUTCOME MEASURES: The major outcome measures were onset and long-term visual field characteristics, disc and peripapillary features, association with other chorioretinal diseases, and mfERG features. RESULTS: Twenty-one eyes had clinical features of chorioretinal syndromes, which are usually associated with an enlarged blind spot. Five eyes were examined too late after onset to expect such features. The visual field defect regressed in all but 12 eyes but never to an unequivocally normal-sized blind spot. Four of the 12 had chorioretinal scarring that corresponded to the permanent field defect. Twenty-one of 26 eyes had peripapillary scarring. The peripapillary scarring appeared the same no matter what the associated chorioretinal disease or type or size of field defect was. mfERG testing of seven patients at follow-up revealed first-order and second order abnormalities long after clinical recovery, abnormalities that were bilateral even when the clinical signs had been unilateral. CONCLUSIONS: If an eye with an enlarged blind spot is examined within 2 weeks of onset, signs of a chorioretinal disease will usually be present. Beyond that period, signs such as disc congestion, disc staining, peripapillary retinitis, foveal changes, and peripheral retinal spots may not be present. Although the patient usually becomes asymptomatic, the blind spot is slightly and permanently enlarged, and there is usually peripapillary disc scarring. mfERG testing indicates that retinal damage is more widespread, bilateral, and permanent than the visual field and clinical features would indicate. Chorioretinal syndromes that are associated with a temporal field defect have some features in common and others that are distinctive. PMID- 12093659 TI - Efficacy and safety of mitomycin-C in primary trabeculectomy: five-year follow up. AB - PURPOSE: To examine the 5-year complications and results of primary trabeculectomy with mitomycin-C (MMC). DESIGN: Retrospective, noncomparative interventional case series. PARTICIPANTS: One hundred twenty-three eyes that underwent primary trabeculectomy with MMC between December 1991 and March 1995. METHODS: All patients received standard trabeculectomy performed by one of two surgeons, using a Weck cell-soaked pledget of MMC, 0.25, 0.33, or 0.5 mg/ml, for 0.5 to 5 minutes. Laser suture lysis was performed postoperatively for intraocular pressure control. MAIN OUTCOME MEASURES: The incidence of complications, including hypotony with or without maculopathy, bleb leak or blebitis, pressure control, and medication reduction at yearly intervals. RESULTS: Mean preoperative and year 5 postoperative intraocular pressures (IOP) were 25.79 and 9.91 mmHg (P < 0.05, paired t test). Hypotony (IOP < 6) occurred in 42.2% of eyes after a mean follow-up of 26.1 months. Hypotony maculopathy occurred in 8.9% of eyes at mean follow-up of 33.7 months. Bleb leak occurred in 14.6% of eyes at a mean follow-up of 27.9 months. Blebitis occurred in 5.7% of eyes at a mean follow-up of 35.4 months, and endophthalmitis occurred in 0.8% of eyes at 15 months; 14.9% of eyes lost 4 lines of visual acuity. The single predictor for the development of late-term hypotony was IOP 1 month after operation (P < 0.05). CONCLUSIONS: Primary mitomycin trabeculectomy significantly lowers IOP at 5 years but is associated with a high incidence of delayed hypotony. PMID- 12093661 TI - Intraoperative episcleral versus postoperative topical application of mitomycin-C for trabeculectomies. AB - OBJECTIVE: To assess the efficacy of a new application technique of mitomycin-C to enhance the outcome of filtering surgery in cases of complicated glaucoma. DESIGN: Randomized clinical trial. PARTICIPANTS: Fifty consecutive patients scheduled for glaucoma surgery in one large surgical center. INTERVENTION: Patients underwent routine trabeculectomy. In group 1, mitomycin-C (0.05 mg/ml) was applied topically to the filtering bleb on days 1, 2, and 3 after surgery (postoperative application). In group 2, mitomycin-C (0.2 mg/ml) was applied by means of to a sponge during surgery (intraoperative application). MAIN OUTCOME MEASURES: Preoperative and postoperative intraocular pressure (IOP) values, visual acuity, the need for antiglaucoma medication, previous surgical procedures, and the need for further surgical interventions were monitored. RESULTS: Sufficient follow-up data were available from 24 of 25 patients in each of the two groups. The follow-up ranged from 12 to 28 months and was evaluated at 12 months for all patients. The mean IOP decreased from 27.3 to 15.5 mmHg in group 1 and from 29.0 to 17.5 mmHg in group 2. The average number of medications decreased from 2.3 and 2.4 to 0.9 and 0.8 (P = 0.68; t test) in groups 1 and 2, respectively, at the 12-month visit. Hypotony was more frequent in group 2, in which the only case of hypotony maculopathy occurred. There was a tendency of more eyes with lower IOP values in this group. The rate of loss of visual acuity of more than 2 lines was higher in group 2. Failures were more frequent in group 2 (7 of 24) compared with group 1 (1 of 24) (P = 0.04, chi-square test). CONCLUSIONS: To our knowledge, this is the first prospective, randomized clinical study to evaluate the efficacy of this different technique of mitomycin-C application. The postoperative application of mitomycin-C was effective, having few failures. This application of mitomycin-C may be associated with a lower rate of complications when used in eyes with complicated forms of glaucoma. PMID- 12093662 TI - Detection of optic neuropathy in glaucomatous eyes with normal standard visual fields using a test battery of short-wavelength automated perimetry and pattern electroretinography. AB - PURPOSE: To evaluate the clinical use of a test battery of short-wavelength automated perimetry (SWAP), frequency-doubling technology (FDT) perimetry, and pattern-electroretinography (PERG) in patients with definite primary open-angle glaucoma (POAG) but normal results on standard automated perimetry (SAP). STUDY DESIGN: Prospective, comparative, observational case series. PARTICIPANTS: Thirty six patients with POAG with standard visual field defects in one eye and normal standard visual fields in the contralateral eye and 36 normal controls were enrolled. MAIN OUTCOME MEASURES: SWAP, PERG, FDT, and SAP were performed in all eyes, and global indices and amplitudes were used for statistical analysis. RESULTS: When contralateral POAG eyes with asymmetric glaucomatous damage was compared, a paired t test showed significant differences in SAP mean deviation (MD) (P < 0.0001), SWAP-MD (P = 0.0003), FDT-MD (P = 0.0008), and PERG amplitudes (P < 0.0001). When comparing between POAG eyes with normal results on SAP and normal controls, Student's t test showed significant differences for SWAP-MD (P < 0.0001), FDT-MD (P = 0.0006), PERG N1P1-amplitude (P = 0.0486) and P1N2-amplitude (P < 0.0001); receiver operating characteristic analysis revealed promising accuracy for SWAP-MD of 73.6% (P < 0.0001). SWAP-MD (P < 0.0001) and FDT-MD (P < 0.0001) correlated significantly with SAP-MD and with each other (range, P < 0.0001 to P = 0.0020). Regression analysis revealed that PERG P1N2-amplitude could improve the power of SWAP-MD from 73.6% to detect early POAG in eyes with normal results on SAP to an accuracy of 81.9%. CONCLUSIONS: A test battery of SWAP-MD and PERG P1N2-amplitude could detect glaucomatous optic neuropathy in POAG eyes with normal standard visual fields, whereas FDT-MD and SWAP-MD significantly correlated with each other and with SAP-MD. All tests were able to detect the eye with the more severe glaucomatous optic neuropathy in patients with asymmetric POAG. PMID- 12093664 TI - Frequency of asymmetric intraocular pressure fluctuations among patients with and without glaucoma. AB - PURPOSE: To describe the frequency and magnitude of asymmetric fluctuations of intraocular pressure (IOP) between fellow eyes of glaucoma patients and normal subjects. DESIGN: Observational case series. PARTICIPANTS: Forty-two subjects without glaucoma and 38 glaucoma patients. METHODS: By means of retrospective chart review, bilateral IOP measurements were examined over time for subjects without glaucoma and for glaucoma patients whose drug regimens remained unchanged during the period included in the study (at least five readings over at least 1 year). Asymmetric IOP fluctuations of at least 3 mmHg and representing at least a 15% change from baseline were identified. MAIN OUTCOME MEASURE: Occurrence of spontaneous asymmetric IOP fluctuations. RESULTS: Twenty-one of 42 subjects without glaucoma exhibited an asymmetric IOP fluctuation between two consecutive visits compared with 24 of 38 glaucoma patients (50% vs. 63.2%, respectively, P = 0.24). Compared with normal subjects, asymmetric IOP fluctuations were observed in 13 of 16 glaucoma patients with prior symmetrical bilateral ocular surgery (50% vs. 81.2%, respectively, P = 0.031), and 11 of 22 ocular surgery-naive glaucoma patients (50% vs. 50%, respectively, P = 1.0). No differences in the magnitudes of the asymmetric IOP fluctuations were noted between groups. Overall, asymmetric IOP fluctuations were observed in 13.7% of follow-up visits among normal subjects versus 16.3% of follow-up visits among glaucoma patients (P = 0.4). CONCLUSIONS: Spontaneous asymmetric fluctuations of intraocular pressure between fellow eyes occur commonly in normal subjects and glaucoma patients. The frequency and magnitude of observed spontaneous asymmetric IOP fluctuations between consecutive clinical visits in glaucoma patients are sufficiently large to potentially confound the interpretation of monocular therapeutic drug trials. PMID- 12093663 TI - Assessment of optic disc anatomy and nerve fiber layer thickness in ocular hypertensive subjects with normal short-wavelength automated perimetry. AB - PURPOSE: To compare optic disc topography and nerve fiber layer thickness in ocular hypertensive eyes and normal subjects. DESIGN: Prospective, case controlled study. PARTICIPANTS AND CONTROLS: One eye in each of 20 normal and 27 ocular hypertensive patients was enrolled. METHODS: Consecutive normal and ocular hypertensive patients were enrolled. Each patient underwent complete ophthalmic examination, achromatic automated perimetry, short-wavelength automated perimetry, confocal scanning laser ophthalmoscopy, confocal scanning laser polarimetry, and optical coherence tomography. The intraocular pressure was 21 mmHg or less for normal subjects and at least 25 mmHg on two separate occasions in ocular hypertensive eyes. Structural parameters were compared between the two groups. Eyes with evidence of glaucomatous optic neuropathy, achromatic visual field loss, or evidence of focal visual field injury during short-wavelength automated perimetry were excluded. MAIN OUTCOME MEASURES: Optic nerve head topography and nerve fiber layer thickness. RESULTS: The three imaging technologies could not detect differences in optic disc or nerve fiber layer anatomy between the two groups. Ocular hypertensive eyes had a greater corrected pattern standard deviation than normal eyes during short-wavelength automated perimetry (P = 0.04). CONCLUSIONS: Ocular hypertensive eyes with normal achromatic automated perimetry and short-wavelength automated perimetry could not be distinguished from normal subjects with confocal scanning laser ophthalmoscopy, confocal scanning laser polarimetry, and optical coherence tomography. PMID- 12093665 TI - Trends in glaucoma surgery incidence and reimbursement for physician services in the Medicare population from 1995 to 1998. AB - OBJECTIVE: To better understand the relationship between glaucoma management and economic incentives, we examined the volume and the reimbursement of argon laser trabeculoplasty (ALT) and trabeculectomy in a 5% random sample of the Medicare population from 1995 to 1998. DESIGN: Retrospective cohort study. PARTICIPANTS: Subjects in a 5% random sample of the Medicare population who had ALT and trabeculectomy from 1995 to 1998. METHODS: Using the Health Care Financing Administration (HCFA) Physician/Supplier Part-B files for a 5% random sample of the Medicare population, we identified all subjects who had ALT and trabeculectomy from 1995 to 1998. Descriptive summaries (the number of surgeries and the mean and the standard deviation of reimbursement per surgery) were calculated for each year. Analysis of variance was used to test for differences in reimbursement per surgery across years. Chi-square tests were used to assess any associations between the changing numbers of ALTs and trabeculectomies over the study period and both age and race. MAIN OUTCOME MEASURES: We assessed the number of ALTs and trabeculectomies and the allowed charges for each surgery in the 5% random sample of the Medicare population from 1995 to 1998. RESULTS: The volume of both ALTs and trabeculectomies declined during the study period. Reimbursement per surgery for both ALT and trabeculectomy varied significantly across years (P < 0.001). Significant associations were found between the changing number of ALTs and both age and race. CONCLUSIONS: Changing numbers of ALT and trabeculectomy seem unrelated to reimbursement rates. Rather, these changes are more likely driven by new developments in the clinical management of glaucoma, among other factors. PMID- 12093666 TI - Recommendations on screening for chloroquine and hydroxychloroquine retinopathy: a report by the American Academy of Ophthalmology. PMID- 12093667 TI - Impaired regulation of HLA-DR expression in human immunodeficiency virus-infected monocytes. PMID- 12093668 TI - Evidence for the presence of immune dysfunction in chronic fatigue syndrome. PMID- 12093670 TI - An aspartyl protease inhibitor orthologue expressed by Parelaphostrongylus tenuis is immunogenic in an atypical host. AB - Parelaphostrongylus tenuis is a neurotropic nematode common in white-tailed deer (Odocoileus virginianus) of eastern North America. This parasite is the causative agent of a debilitating neurologic disease in atypical hosts, including domestic livestock. In order to identify proteins of potential significance in the host parasite relationship, a cDNA library was produced from adult P. tenuis mRNA. Screening the library with antisera from infected red deer (Cervus elaphus elaphus) and immunized AO strain rats, we identified clones with sequence similarities to aspartyl protease inhibitors from several parasitic nematodes. Antibody that was generated against this recombinant protein of P. tenuis (Pt-API 1) detected the native protein in E/S products, in muscle and gonad, and on the surface of the cuticle of adult male and female P. tenuis. The native protein was detected in internal structures of first-stage (L1) and third-stage (L3) larvae. Reverse transcription-PCR confirmed expression of Pt-api-1 in L1, L3, and adult male and female worms. Expression of Pt-API-1 throughout the life cycle of P. tenuis suggests an essential function. Antibodies specific for recombinant Pt-API 1 were detected by enzyme-linked immunosorbent assay in sera from 12 red deer experimentally infected with P. tenuis. Antibodies were detected within 28 to 56 days postinfection. Responses were sustained or biphasic in animals with patent infections, consistent with expression of Pt-API-1 by L1. Our results are compatible with findings in other parasitic nematodes showing that aspartyl protease inhibitors are highly immunogenic. PMID- 12093669 TI - Evaluation of events occurring at mucosal surfaces: techniques used to collect and analyze mucosal secretions and cells. PMID- 12093671 TI - Responses of adipose and muscle lipoprotein lipase to chronic infection and subsequent acute lipopolysaccharide challenge. AB - Infection of male Swiss Webster mice with Toxoplasma gondii or Neospora caninum leads to long-term alterations in energy balance. Following an initial 20 to 30% weight loss in all T. gondii-infected mice, half of the animals regain most of the lost weight (gainers), whereas the others maintain their low body weight (nongainers). Infection with N. caninum does not elicit weight loss. Lipoprotein lipase (LPL), the enzyme responsible for plasma triglyceride (TG) clearance and partitioning among tissues, is under tissue-specific modulation associated with energy balance. It is also a major determinant of infection-induced hypertriglyceridemia. This study aimed to assess the long-term modulation of adipose and muscle LPL activity in mice infected with T. gondii or N. caninum, to evaluate the effects of subsequent acute lipopolysaccharide (LPS) administration, and to relate LPL modulation in these conditions with infection-related changes in body weight gain. Twenty-eight days after infection, LPL activity in muscle of both gainer and nongainer T. gondii-infected mice was reduced by 40 to 50% compared with the levels in controls and N. caninum-infected mice, whereas LPL activity in adipose depots remained unchanged in all infected groups compared to the level in controls. LPS (from Escherichia coli, 100 ng/kg) injection induced a global reduction in adipose LPL in all groups, as assessed 90 min later. In both T. gondii-infected subgroups, muscle LPL was not further reduced by LPS treatment, whereas it was decreased by 40 to 50% in muscles of control and N. caninum-infected mice. Pre-LPS TG levels in plasma were similar in all groups. LPS greatly increased TG levels in plasma in both control and N. caninum-infected animals, whereas it did not alter those of T. gondii-infected gainer or nongainer animals. These results show that (i) independently of the extent of postinfection weight gain, long-term infection with T. gondii chronically reduces muscle LPL, which becomes unresponsive to acute endotoxemia; (ii) modulation of tissue LPL activity during chronic T. gondii infection favors TG partitioning towards adipose tissue; and (iii) skeletal muscle LPL is a key determinant of the acute response of triglyceridemia to LPS. PMID- 12093673 TI - Catalase, a specific antigen in the feces of human subjects infected with Helicobacter pylori. AB - Recently, we reported the production of three new monoclonal antibodies with high specificity for a Helicobacter pylori antigen suitable for diagnosis of H. pylori infection. The aim of the present study was to identify the antigen recognized by these monoclonal antibodies concerning both H. pylori and the feces of human subjects infected with H. pylori. The cellular antigen was purified from an H. pylori cell extract by immunoaffinity column chromatography with the monoclonal antibody as a ligand. The amino-terminal amino acid sequences (eight residues) of the purified antigen and H. pylori catalase were the same. The molecular weights of native and subunit, specific catalase activity, and UV and visible spectra of the purified antigen were in good agreement with those of H. pylori catalase. The human fecal antigens were purified from two fecal samples of two H. pylori positive subjects by ammonium sulfate precipitation, CM-Sephadex C(50) chromatography, and the same immunoaffinity chromatography used for the H. pylori cellular antigen. The fecal antigens had catalase activity. The amino-terminal amino acid sequences (five residues) of the human fecal antigen and H. pylori catalase were the same. The monoclonal antibodies reacted with the native cellular antigen, but did not react with the denatured antigen, human catalase, and bovine catalase. The results show that the target antigen of the monoclonal antibodies is native H. pylori catalase and that the monoclonal antibodies are able to specifically detect the antigen, which exists in an intact form, retaining the catalase activity in human feces. PMID- 12093672 TI - Interleukin-18 induces acute biphasic reduction in the levels of circulating leukocytes in mice. AB - We investigated the acute hematological changes caused by interleukin-18 (IL-18) in mice. Intraperitoneal administration of IL-18 (2 microg/mouse) resulted in biphasic decreases in the number of leukocytes in the blood. The first phase of decrease occurred within 2 h of IL-18 administration and was followed by a transient increase at 5 h. The second phase of decrease occurred at around 6 h, reaching a nadir which lasted for more than 24 h. In mice deficient in inducible nitric oxide (NO) synthase, the first phase of reduction of leukocytes did not occur although the second phase of decrease was observed. In mice deficient in gamma interferon (IFN-gamma) or in mice depleted of natural killer cells and incapable of producing IFN-gamma, IL-18 had no effect on the number of circulating leukocytes. Levels of nitrite and/or nitrate in the serum were elevated within 2 h after administration of IL-18, reaching a peak at 4 h and then decreasing gradually to the basal level over a 24-h period of time. On the other hand, serum IFN-gamma levels changed in a biphasic manner, reaching a peak at 2 h after IL-18 administration, followed by a decrease in the basal level and a second increase at 6 h. Levels of IL-18 receptor mRNAs also showed biphasic changes in correlation with the changes in serum IFN-gamma levels. These results suggest that the changes in the leukocyte number following IL-18 administration are mediated by NO and IFN-gamma, with NO being involved in the first phase of reduction and IFN-gamma being involved in both phases. PMID- 12093675 TI - Value of microimmunofluorescence for diagnosis and follow-up of Bartonella endocarditis. AB - Bartonella endocarditis is a disease of emerging importance that causes serious complications and high rates of mortality. Due to the fastidious nature of Bartonella species and their high degrees of antibiotic susceptibility, cultures of clinical samples most often remain sterile and valvular biopsy specimens, the best specimens for PCR amplification, are seldom available. Therefore, serology appears to be the easiest diagnostic tool. In order to determine the best cutoff value for serology and its predictive values for the detection of Bartonella endocarditis, we studied 48 patients with culture- and/or PCR-confirmed Bartonella endocarditis. We also applied these serological criteria to 156 patients with blood culture-negative endocarditis. Furthermore, we compared the kinetics of the antibody responses to Bartonella spp. in order to estimate the value of serology for prediction of the occurrence of relapses. A titer of > or = 1:800 for immunoglobulin G antibodies to either Bartonella henselae or B. quintana has a positive predictive value of 0.810 for the detection of chronic Bartonella infections in the general population and a value of 0.955 for the detection of Bartonella infections among patients with endocarditis. When this cutoff was applied to 156 patients with blood culture-negative endocarditis, we were able to diagnose Bartonella infections in an additional 45 patients with definite endocarditis for whom a positive Bartonella serology was the only evidence of infection. On follow-up, the kinetics of the decrease in antibody titers were significantly delayed in two patients with relapses. In conclusion, we recommend the determination of antibodies to both B. quintana and B. henselae and the use of a cutoff value of 1:800 for the diagnosis of Bartonella endocarditis. We propose that this criterion, which may also help with the detection of late relapses, be included as a major criterion in the Duke criteria for the diagnosis of infective endocarditis. PMID- 12093674 TI - Diagnosis of visceral leishmaniasis by enzyme-linked immunosorbent assay using urine samples. AB - A diagnostic method has been developed to detect anti-Leishmania donovani immunoglobulin G (IgG) in urine by enzyme-linked immunosorbent assay (ELISA). In measuring anti-L. donovani IgG, IgA, and IgM in urine, the method performed best in the detection of IgG. The sensitivity and specificity of the assay were determined with panels of urine samples from 62 visceral leishmaniasis (VL) patients, 59 healthy controls from areas of endemicity, 53 healthy controls from areas of nonendemicity, 59 malaria patients, 13 tuberculosis patients, 23 cutaneous leishmaniasis patients, and 7 patients with other diseases. Using L. donovani promastigote crude antigen, the test had 93.5% sensitivity (58 positives of 62 VL patient samples) and 89.3% specificity (191 negatives of 214 non-VL patient samples). The ELISA with acetone-treated L. donovani promastigote antigen raised the sensitivity and specificity to 95.0 and 95.3%, respectively. Western blot analysis revealed that most of the samples that cross-reacted with crude antigen in ELISA did not recognize any antigenic component of L. donovani crude antigen. We also checked 40 serum samples from the same group of VL patients for anti-L. donovani IgG and got 90.0% sensitivity with both crude and acetone treated antigens. As collection of urine is much easier than collection of serum, the detection of anti-L. donovani IgG in urine with acetone-treated antigen will be useful in epidemiological studies. It could be an adjunct of laboratory diagnosis. PMID- 12093676 TI - Regulation of interleukin-8 by interleukin-10 and transforming growth factor beta in human monocytes infected with mycobacterium bovis. AB - Recent studies indicate that interleukin 8 (IL-8) production contributes to the host immune responses against mycobacterial infection. In this study, we were interested to determine whether induction of IL-8 in human monocytes infected with Mycobacterium bovis was regulated by other monocyte-derived cytokines important in antimycobacterial immunity: IL-10 and transforming growth factor beta (TGF-beta). Here, we report that IL-10 reduced, in a graded and significant manner, IL-8 production by M. bovis-infected human monocytes. Additionally, the specificity of the observed inhibition was further confirmed, since the addition of an anti-IL-10 neutralizing antibody completely reversed the inhibitory effect. In contrast, addition or neutralization of TGF-beta appeared to have no significant effect on M. bovis-induced IL-8 secretion by human monocytes, whereas CD40 expression on M. bovis-infected monocytes was significantly inhibited by this cytokine. This was consistent with the finding by the reverse transcription PCR method that pretreatment with IL-10, but not TGF-beta, potently inhibited IL 8 mRNA levels. Interestingly, neutralization of endogenous IL-10 did not significantly alter IL-8 secretion, suggesting that induction of IL-8 was not significantly affected by coexpression of IL-10 during infection of human monocytes with M. bovis. Collectively, these data indicate that IL-8 production may be regulated when human monocytes are exposed to IL-10 prior to activation with M. bovis BCG. These data will aid in our understanding of the mechanisms involved in regulating the protective immune response to stimulation with M. bovis BCG. PMID- 12093677 TI - Antigenicity of Leishmania braziliensis histone H1 during cutaneous leishmaniasis: localization of antigenic determinants. AB - The humoral immune response against Leishmania braziliensis histone H1 by patients with cutaneous leishmaniasis is described. For this purpose, the protein was purified as a recombinant protein in a prokaryotic expression system and was assayed by enzyme-linked immunosorbent assay (ELISA) with a collection of sera from patients with cutaneous leishmaniasis and Chagas' disease. The assays showed that L. braziliensis histone H1 was recognized by 66% of the serum samples from patients with leishmaniasis and by 40% of the serum samples from patients with Chagas' disease, indicating that it acts as an immunogen during cutaneous leishmaniasis. In order to locate the linear antigenic determinants of this protein, a collection of synthetic peptides covering the L. braziliensis histone H1sequence was tested by ELISA. The experiments showed that the main antigenic determinant is located in the central region of this protein. Our results show that the recombinant L. braziliensis histone H1 is recognized by a significant percentage of serum samples from patients with cutaneous leishmaniasis, but use of this protein as a tool for the diagnosis of cutaneous leishmaniasis is hampered by the cross-reaction with sera from patients with Chagas' disease. PMID- 12093678 TI - Enzymatic activities of bovine peripheral blood leukocytes and milk polymorphonuclear neutrophils during intramammary inflammation caused by lipopolysaccharide. AB - Leukocytes are recruited from peripheral blood into milk as part of the inflammatory response to mastitis. However, excessive accumulation of inflammatory cells alters the quality of milk and the proteases produced by polymorphonuclear neutrophils (PMNs) and macrophages may lead to mammary tissue damage. To investigate PMN recruitment and the kinetics of their intracytoplasmic enzymes in inflammation, we generated mastitis in six cows by intramammary infusion of lipopolysaccharide (LPS). Clinical signs of acute mastitis were observed in all of the cows, and normal status was resumed by 316 h. Intracytoplasmic elastase, collagenase, and cathepsin activities were measured within live cells by flow cytometry in peripheral blood leukocytes and milk PMNs before and during the inflammatory process (at 10 time points between 4 and 316 h). The proportion of immature PMNs was appreciated by CD33 surface labeling measured in flow cytometry. Leukopenia was observed in the peripheral blood 4 h postinfusion, concomitant to an increase in somatic cell counts in milk. CD33(+) PMNs were preferentially recruited from the peripheral blood to milk. Enzymatic activities were detected in PMNs, lymphocytes, and monocytes at levels depending on the cell type, sample nature, and time of collection. Milk PMNs had lower enzymatic activities than peripheral blood PMNs. This study showed that milk PMNs recruited during LPS-induced experimental mastitis have an immature phenotype and significantly lower enzymatic activities than peripheral blood PMNs. This suggests that CD33, an adhesion molecule, may be involved in the egress from blood to milk and that the enzymatic contents of PMNs are partly used during this process. PMID- 12093680 TI - Validation of an in-house assay for cytomegalovirus immunoglobulin G (CMV IgG) avidity and relationship of avidity to CMV IgM levels. AB - Measurement of cytomegalovirus (CMV)-specific immunoglobulin G (IgG) avidity has proven to be a powerful tool for distinguishing primary from nonprimary CMV infection. An in-house enzyme-linked immunosorbent assay (ELISA) for measuring CMV IgG avidity was validated using 84 sera from pregnant women who had recently seroconverted following primary CMV infection and 74 sera from individuals with past CMV infection (IgG-positive and IgM-negative profile). Of the 84 sera from pregnant women, 73 sera were collected within 120 days of the last IgG-negative sample, and 72 of these 73 sera (99%) exhibited an avidity index (AI) of <50%. In contrast, 71 of 74 (96%) sera from individuals with past CMV infection exhibited CMV AI values of > 60%. Thus, low avidity in the in-house ELISA was defined as an AI of < or = 50%, whereas high avidity was defined as an AI of > or = 60%. In additional studies, the relationship between CMV IgG avidity and CMV IgM levels was examined using 64 CMV IgG-positive sera (time since seroconversion unknown) exhibiting equivocal or positive results in a CMV IgM capture ELISA (Diamedix). Of these 64 sera, 29 exhibited IgM index values of > or = 3.0, and 27 of these 29 (93%) exhibited low IgG avidity. A similar trend was observed when a subset of these 64 sera (n = 48) was tested in another CMV IgM capture ELISA (Trinity); of 18 sera with IgM index values of > or = 3.0, 17 (94%) exhibited low IgG avidity. These findings demonstrate the validity of an in-house ELISA for CMV IgG avidity and further show that strong reactivity of CMV IgG-positive sera in either of two CMV IgM capture assays is a reliable indicator of low CMV IgG avidity, and thus, recent CMV infection. PMID- 12093681 TI - Genetic heterogeneity of small ruminant lentiviruses involves immunodominant epitope of capsid antigen and affects sensitivity of single-strain-based immunoassay. AB - The pol and gag gene fragments of small ruminant lentivirus field isolates collected in the last decade in Italy were amplified, sequenced, and analyzed. Phylogenetic analysis revealed that the majority of ovine isolates form a distinct cluster more similar to caprine lentivirus prototypes than to the visna virus prototype. These findings confirm and extend those reported by Leroux et al. (Arch. Virol., 142:1125-1137, 1997). Moreover, we observed that a variable region of Gag, included in the fragment analyzed, corresponded to one of the three major capsid antigen epitopes, which suggests that the antibody response to this epitope may be type specific. To test this hypothesis, two recombinant peptides, derived from the Icelandic prototype K1514 and this novel genotype, were expressed and used in an enzyme-linked immunosorbent assay to screen a panel of ovine and caprine sera collected from different geographical locations in Italy. Several sera reacted in a type-specific manner, indicating that in a diagnostic setting the combination of at least these two type-specific peptides is necessary to cover a wide range of infections. Additionally, these results support the hypothesis of cross-species transmission based on the phylogenetic analysis described above. This has implications for the control and eradication of small ruminant lentivirus infections. PMID- 12093679 TI - Sputum cytokine levels in patients with pulmonary tuberculosis as early markers of mycobacterial clearance. AB - Sputum and serum from patients with active pulmonary tuberculosis (TB), healthy purified protein derivative-positive adults, and patients with bacterial pneumonia were collected to simultaneously assess local immunity in the lungs and peripheral blood. To determine whether cytokine profiles in sputum from TB patients and control subjects were a reflection of its cellular composition, cytospin slides were prepared in parallel and assessed for the presence of relative proportions of epithelial cells, neutrophils, macrophages, and T cells. Gamma interferon (IFN-gamma) in sputum from TB patients was markedly elevated over levels for both control groups. With anti-TB therapy, IFN-gamma levels in sputum from TB patients decreased rapidly and by week 4 of treatment were comparable to those in sputum from controls. Further, IFN-gamma levels in sputum closely followed mycobacterial clearance. Although detected at fourfold-lower levels, IFN-gamma immunoreactivities in serum followed kinetics in sputum. TNF alpha, interleukin 8 (IL-8) and IL-6 also were readily detected in sputum from TB patients at baseline and responded to anti-TB therapy. In contrast to IFN-gamma, however, TNF-alpha and IL-8 levels also were elevated in sputum from pneumonia controls. These data indicate that sputum cytokines correlate with disease activity during active TB of the lung and may serve as potential early markers for sputum conversion and response to anti-TB therapy. PMID- 12093682 TI - Performance of three microimmunofluorescence assays for detection of Chlamydia pneumoniae immunoglobulin M, G, and A antibodies. AB - The microimmunofluorescence (MIF) test is considered the "gold standard" for laboratory diagnosis of acute and chronic Chlamydia pneumoniae infection. The performance of a MIF test based on C. pneumoniae antigen from Washington Research Foundation (WRF) was compared with those of assays from Labsystems (LAB) and MRL Diagnostics (MRL) by investigation of sera from three groups of patients: group I, 83 sera from 28 patients with atypical pneumonia; group II, 37 sera from 16 patients with acute C. pneumoniae or Chlamydia psittaci respiratory tract infection confirmed by PCR or culture; group III, 100 sera from 100 persons enrolled in the Copenhagen City Heart Study. The accordance among the results of the WRF assay and the two commercial assays was excellent for the immunoglobulin M (IgM) antibody detection rate (98%). The accordance in detection rates for IgG and IgA antibodies in sera from patients with acute infections was acceptable (87 and 88%), and in sera from group III, it was excellent (95 and 97%). The determinations of endpoint titers were reproducible with <1 dilution step difference for all three methods, except that the mean IgM antibody titer found by the LAB assay was almost 2 dilution steps higher than that found by the other two methods. Although the three assays use different C. pneumoniae strains as antigens, the detection rates and IgG and IgA endpoint titers were similar. The difference in endpoint titers of IgM antibodies is of no major concern, as the diagnosis of acute C. pneumoniae infection rests on the presence of IgM antibodies, not on their level. PMID- 12093683 TI - Evaluation of urine as a clinical specimen for diagnosis of hepatitis a. AB - The present study pertains to the evaluation of urine as a specimen for detection of anti-hepatitis A virus (anti-HAV) antibodies. Immunoglobulin M (IgM), IgG, and IgA capture enzyme-linked immunosorbent assays for hepatitis A were performed on paired serum and urine specimens collected from hepatitis A patients (n = 92), healthy individuals (n = 100), non-A hepatitis patients (n = 70), and patients with nonhepatic diseases (n = 64, including 37 renal disease patients). Hepatitis A patients seropositive for anti-HAV IgM showed 95.65% uropositivity. No false positive reactions were observed in control groups. The uropositivity of anti-HAV IgM persisted during the convalescent phase of the disease. Anti-HAV IgG uropositivity correlated well with corresponding seropositivity in all groups (P > 0.05 for each). No significant difference between the proportions of serum and urine positivity for anti-HAV IgA was noted (P > 0.05 for each). Using seroreactivity as a "gold standard," the sensitivity and specificity for anti-HAV IgM, anti-HAV IgG, and anti-HAV IgA tests with urine as a specimen were found to be 95.65 and 100%, 97.76 and 76.47%, and 92.23 and 88.18%, respectively. Urine appears to be comparable to serum for diagnosis of recent and past infection with hepatitis A. PMID- 12093684 TI - Identification, molecular cloning, and evaluation of potential use of isocitrate dehydrogenase II of Mycobacterium bovis BCG in serodiagnosis of tuberculosis. AB - Diagnosis of tuberculosis is time-consuming and requires infrastructures which are often not available in countries with high incidences of the disease. In the present study, an 82-kDa protein antigen was isolated by affinity chromatography and was identified by peptide mass fingerprinting as isocitrate dehydrogenase II, which is encoded by the icd2 gene of Mycobacterium bovis BCG. The icd2 gene of BCG was cloned by PCR, and the product of recombinant gene expression was purified and analyzed by two-dimensional polyacrylamide gel electrophoresis. The recombinant protein, named rICD2, was tested for its recognition by immunoglobulin G (IgG) antibodies from the sera of 16 patients with tuberculosis (TB) and 23 healthy individuals by Western blotting. The results showed that rICD2 is recognized by IgG antibodies from the sera of all TB patients tested at serum dilutions of > or = 1:640. At a serum dilution of 1:1,280, the sensitivity was 50% and the specificity was 86.9%. These results indicate that rICD2 might represent a candidate for use in a new assay for the serodiagnosis of TB. PMID- 12093685 TI - Effects of psychological stress and alprazolam on development of oral candidiasis in rats. AB - Psychological stress has been found to suppress cell-mediated immune responses that are important in limiting the proliferation of Candida albicans. Since anxiolytic drugs can restore cellular immunity in rodents exposed to stress conditions, we designed experiments conducted to evaluate the effects of alprazolam (1 mg/kg of body weight/day), a central benzodiazepine anxiolytic agonist, on the development of oral candidiasis in Sprague-Dawley rats exposed to a chronic auditory stressor. Animals were submitted to surgical hyposalivation in order to facilitate the establishment and persistence of C. albicans infection. Application of stress and treatment with drugs (placebo or alprazolam) were initiated 7 days before C. albicans inoculation and lasted until the end of the experiments (day 15 postinoculation). Establishment of C. albicans infection was evaluated by swabbing the inoculated oral cavity with a sterile cotton applicator on days 2 and 15 after inoculation, followed by plating on YEPD (yeast extract peptone-dextrose) agar. Tissue injury was determined by the quantification of the number and type (normal or abnormal) of papillae on the dorsal tongue per microscopic field. A semiquantitative scale was devised to assess the degree of colonization of the epithelium by fungal hyphae. Our results show that stress exacerbates C. albicans infection of the tongues of rats. Significant increases in Candida counts, the percentage of the tongue's surface covered with clinical lesions, the percentage of abnormal papillae, and the colonization of the epithelium by fungal hyphae were found in stressed rats compared to those found in the unstressed rats. Treatment with alprazolam significantly reversed these adverse effects of stress, showing that, besides the psychopharmacological properties of this anxiolytic drug against stress, it has consequences for Candida infection. PMID- 12093686 TI - MaGiK method of T-Cell receptor repertoire analysis. AB - T-cell receptor diversity enables the cellular immune response to recognize a broad range of viral and other pathogenic agents. An increasingly common method of characterizing T-cell receptor diversity and usage in response to antigenic challenges involves the identification of clonal expansions by PCR amplification of the CDR3 region of distinct TCRVbeta families. Though clonal expansions often appear evident upon visual inspection of the results, a systematic method is needed for the valid enumeration of these expansions. Here, we describe a novel analysis method, termed the MaGiK method, for systematically identifying and enumerating clonal T-cell expansions and for applying the results to investigations of the T-cell receptor repertoire. PMID- 12093687 TI - Epigallocatechin gallate, a potential immunomodulatory agent of tea components, diminishes cigarette smoke condensate-induced suppression of anti-Legionella pneumophila activity and cytokine responses of alveolar macrophages. AB - Even though cigarette smoking has been shown to suppress immune responses in the lungs, little is known about the effect of cigarette smoke components on respiratory infections. In the present study, the effects of cigarette smoke condensate (CSC) on bacterial replication in alveolar macrophages and the immune responses of macrophages to infection were examined. Furthermore, a possible immunotherapeutic effect of epigallocatechin gallate (EGCg), a major form of tea catechins, on the CSC-induced suppression of antimicrobial activity and immune responses of alveolar macrophages was also determined. The treatment of murine alveolar macrophage cell line (MH-S) cells with CSC significantly enhanced the replication of Legionella pneumophila in macrophages and selectively down regulated the production of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) induced by bacterial infection. The treatment of macrophages with EGCg not only overcame the CSC-induced suppression of antimicrobial activity but also strengthened the resistance of macrophages to infection. EGCg also markedly up-regulated the CSC-suppressed IL-6 and TNF-alpha production by macrophages in response to infection. The results of exogenous TNF-alpha treatment and neutralization treatment with anti-TNF-alpha and anti-gamma-interferon (IFN gamma) antibodies and the determination of IFN-gamma mRNA levels indicate that CSC-suppressed macrophages can be activated by EGCg to inhibit L. pneumophila growth by up-regulation of TNF-alpha and IFN-gamma production. Thus, this study revealed that CSC selectively alters the immune responses of macrophages to L. pneumophila infection and leads to an enhancement of bacterial replication in macrophages. In addition, the tea catechin EGCg can diminish such suppressive effects of CSC on alveolar macrophages. PMID- 12093688 TI - Comparison of a multiplex flow cytometric assay with enzyme-linked immunosorbent assay for auantitation of antibodies to tetanus, diphtheria, and Haemophilus influenzae Type b. AB - We developed a multiplexed indirect immunofluorescence assay for antibodies to Haemophilus influenza type b (Hib) polysaccharide and the toxoids of Clostridium tetani (Tet) and Corynebacterium diphtheriae (Dip) based on the Luminex multiple analyte profiling system. A pooled serum standard was calibrated against World Health Organization standards for Dip and Tet and an international standard for Hib. The multiplexed Luminex assay was compared to individual enzyme-linked immunosorbent assays (ELISAs) for the same analytes. By both methods, 75 (92.6%) of 81 of random serum samples had protective levels of antibody to Tet (> or = 0.1 IU/ml). For Dip, 81.5% of the samples had protective antibody levels (> or = 0.1 IU/ml) by ELISA and 80.2% had protective antibody levels by Luminex. Protective levels (> or = 1.0 microg/ml) of antibody to Hib were found in 45.0% of the samples tested by ELISA and in 39.0% of the samples tested by Luminex. The correlations (R(2)) between ELISA and Luminex of the 81 samples were 0.96, 0.96, and 0.91 for Tet, Dip, and Hib, respectively. There was also similar agreement between Luminex and ELISA for sera collected before and 1 month after Tet, Dip, and Hib vaccine administration. Both methods detected strong postvaccination responses. The Luminex method is an attractive alternative to ELISA since it reduces labor and reagent costs, as well as assay time. PMID- 12093690 TI - Use of multiepitope polyproteins in serodiagnosis of active tuberculosis. AB - Screening of genomic expression libraries from Mycobacterium tuberculosis with sera from tuberculosis (TB) patients or rabbit antiserum to M. tuberculosis led to the identification of novel antigens capable of detecting specific antibodies to M. tuberculosis. Three antigens, Mtb11 (also known as CFP-10), Mtb8, and Mtb48, were tested together with the previously reported 38-kDa protein, in an enzyme-linked immunosorbent assay (ELISA) to detect antibodies in TB patients. These four proteins were also produced as a genetically fused polyprotein, which was tested with two additional antigens, DPEP (also known as MPT32) and Mtb81. Sera from individuals with pulmonary and extrapulmonary TB, human immunodeficiency virus (HIV)-TB coinfections, and purified protein derivative (PPD)-positive and PPD-negative status with no evidence of disease were tested. In samples from HIV-negative individuals, the ELISA detected antibodies in >80% of smear-positive individuals and >60% smear-negative individuals, with a specificity of approximately 98%. For this group, smears detected 81.6% but a combination of smear and ELISA had a sensitivity of approximately 93%. The antigen combination detected a significant number of HIV-TB coinfections as well as antibodies in patients with extrapulmonary infections. Improved reactivity in the HIV-TB group was observed by including the antigen Mtb81 that was identified by proteomics. The data indicate that the use of multiple antigens, some of which are in a single polyprotein, can be used to facilitate the development of a highly sensitive test for M. tuberculosis antibody detection. PMID- 12093689 TI - Time course of humoral and cell-mediated immune responses to human papillomavirus type 16 in infected women. AB - The time course of cell-mediated and humoral immune responses was elucidated in eight women with human papillomavirus type 16 (HPV-16) infection by performing serial HPV-16 E6 and E7 cytotoxic T-lymphocyte (CTL) assays and HPV-16 virus-like particle (VLP) antibody analyses. Four subjects had a single incident of HPV-16 DNA detection, and four subjects had two periods of HPV-16 DNA detection. In two of the women in the latter group, the second episode of HPV-16 detection occurred in the presence of high titers of HPV-16 VLP antibody, bringing into question the protective role of humoral immunity in preventing repeated infection. However, all four subjects rapidly became HPV-16 DNA negative following the second detection of HPV-16 DNA, suggesting the presence of immunological memory. In addition, one subject rapidly became negative for HPV-16 DNA despite having no evidence of CTL or VLP antibody response prior to the second HPV-16 DNA detection, suggesting the presence of immunological responses at an undetectable level. Overall, seven of eight subjects (88%) had detectable HPV-16 E6 and/or E7 CTL responses and seven of eight women (88%) had detectable HPV-16 VLP antibody responses. PMID- 12093691 TI - Induction of cytokines by glucosyltransferases of streptococcus mutans. AB - Production of proinflammatory cytokines is implicated in the pathogenesis of viridans streptococcus-induced alpha-streptococcal shock syndrome and infective endocarditis. Streptococcus mutans, one of the opportunistic pathogens causing infective endocarditis, was reported previously to stimulate monocytes and epithelial and endothelial cells in vitro to produce various cytokines. We found that glucosyltransferases (GTFs) GtfC and GtfD of S. mutans stimulated predominantly the production of interleukin-6 (IL-6) from T cells cultured in vitro. The level of IL-6 but not of tumor necrosis factor alpha in blood was significantly elevated when rats were injected intravenously with S. mutans GS-5, whereas IL-6 was detected at a much lower level when rats were challenged with NHS1DD, an isogenic mutant defective in the expression of GTFs. The serum IL-6 level was elevated in patients with endocarditis caused by different species of viridans streptococci which express GTF homologues. Affinity column-purified GTFs reduced the levels of detectable IL-2 of T cells stimulated by another bacterial antigen, tetanus toxoid. These results suggested that GTFs might modulate the production of Th1-type cytokines and that GTFs of S. mutans play a significant role in stimulating the production of the proinflammatory cytokine IL-6 in vivo. PMID- 12093692 TI - Screening of neonatal calves for persistent infection with bovine viral diarrhea virus by immunohistochemistry on skin biopsy samples. AB - Detection and elimination of cattle that are persistently infected with bovine viral diarrhea virus (BVDV) is important for controlling the transmission of this virus. Colostrum-derived antibodies make the detection of persistently BVDV infected neonatal calves cumbersome and expensive. The objective of this study was to evaluate the use of immunohistochemical staining of skin biopsy samples from neonatal calves as a method for the early detection of persistent BVDV infection. Three hundred thirty-two 1- to 4-week-old dairy calves were screened for BVDV as part of a routine control program. Formalin-fixed skin biopsy samples were stained for BVDV antigen by immunohistochemistry (IHC), and the results were compared to those of virus isolation (VI) from white blood cell preparations. Six calves were positive by both IHC and VI and remained positive for BVDV upon subsequent follow-up testing; thus, they were classified as persistently infected with BVDV. One calf was positive by VI but negative by IHC. On subsequent testing, the calf was negative by VI, suggesting that the initial VI result was due to an acute BVDV infection. One calf was positive by IHC but negative by VI. This calf remained negative by VI on follow-up testing. Immunohistochemical staining of skin biopsy samples is a reliable method for screening neonatal calves for persistent BVDV infection and would be a useful management tool as an aid for controlling and preventing BVDV infection. PMID- 12093693 TI - Bactericidal activity in whole blood as a potential surrogate marker of immunity after vaccination against tuberculosis. AB - The development of new tuberculosis (TB) vaccines will require the identification of correlates of human protection. This study examined the balance between immunity and virulence in a whole blood infection model in which intracellular mycobacterial survival was measured using BACTEC. In the blood of tuberculin negative donors, counts of Mycobacterium tuberculosis H(37)Ra organisms fell by 0.14 log(10) CFU during 96 h of whole blood culture, whereas counts of Mycobacterium bovis BCG, M. tuberculosis H(37)Rv, and a clinical TB isolate's organisms increased by 0.13, 0.43, and 1.04 log(10) CFU, respectively (P < 0.001), consistent with their relative virulence. Inhibition of tumor necrosis factor alpha by the addition of methylprednisolone or pentoxifylline or removal of CD4(+) or CD8(+) T cells by magnetic beads had deleterious effects on immune control of intracellular growth only in the blood of tuberculin-positive donors. Repeated vaccination of eight tuberculin-negative volunteers with M. bovis BCG resulted in a 0.3 log (50%) reduction in BCG CFU counts in the model compared to baseline values (P < 0.05). Three of the volunteers responded only after the second vaccination. These experiments indicate that whole blood culture may be used to measure immunity to M. tuberculosis and that further studies of repeated BCG vaccination are warranted. PMID- 12093694 TI - Ability of the borreliacidal antibody test to confirm lyme disease in clinical practice. AB - Highly specific borreliacidal antibodies are induced by infection with Borrelia burgdorferi, and a borreliacidal antibody test (BAT) may be an accurate laboratory procedure for confirming Lyme disease in clinical practice. To investigate this, 34 Lyme disease sera and 34 sera from patients with other illnesses who had presented to a primary-care facility located in an area of borreliosis endemicity were tested by the BAT and Western blotting (WB). The BAT was more sensitive (79% versus 65%; P = 0.090), especially in cases in which patients had a single erythema migrans lesion (P = 0.021). In addition, the potentially cross-reactive sera were negative by the BAT but WB yielded three (9%) false-positive results. The results from 104 sera from possible Lyme disease patients demonstrated the clinical usefulness of the more sensitive and specific BAT. The BAT was positive for 40 (38%) sera from patients with Lyme disease related symptoms and appropriate clinical and epidemiological findings. WB confirmed Lyme disease in 30 (75%) of the 40 BAT-positive patients but failed to detect B. burgdorferi infection in 10 BAT-positive patients. WB was also positive for 11 BAT-negative sera, but six (55%) patients had case histories which suggested that the results were false positives. Collectively, the results confirm that the BAT is a sensitive and highly specific test and suggest that widespread use would increase the accuracy of serodiagnostic confirmation of Lyme disease. PMID- 12093695 TI - Preadult stage parasites and multiple timed exposure to infective larvae are involved in development of limb edema in Brugia malayi-infected Indian leaf monkeys (Presbytis entellus). AB - The pathogenesis of filarial limb edema is not known. The role of parasitological variables and parasite-mediated phenomena in the development of limb edema was investigated in the Presbytis entellus-Brugia malayi model. Infection was initiated with subcutaneous inoculation of infective third-stage larvae (L(3)), and the animals were reexposed to different doses of L(3) at the prepatent, patent, and diminishing microfilaremia (0 to 5% of peak microfilaremia count) stages of infection. A large L(3) inoculum size and repeated inoculation in the ankle region during the prepatent, patent, and diminishing microfilaremia stages of infection were found to be necessary for reproducible induction of limb edema. The preadult stage of the parasite was found to be the most potent inducer of limb edema, followed by L(5) and L(4). The presence of the proinflammatory cytokines tumor necrosis factor alpha, interleukin-1beta, and interleukin-6 in edema fluid in the leg receiving the parasite challenge indicated that the limb edema development was due to parasite-mediated cytokine responses. The absence of bacterial infection or anti-streptolysin O titer in the edema fluid and blood indicated that bacterial infection is not necessary for the development of limb edema. PMID- 12093696 TI - In vitro assessment of antiborrelial activity of OspA vaccine sera. AB - Prevention of Lyme disease by the recombinant OspA-based vaccine reportedly works by preventing transmission of spirochetes from ticks to humans. We report on an in vitro microculture assay, which can be used to provide an indicator of the need for booster doses of vaccine. PMID- 12093697 TI - Complement-mediated neutralization of canine distemper virus in vitro: cross reaction between vaccine Onderstepoort and field KDK-1 strains with different hemagglutinin gene characteristics. AB - The properties of neutralization of antigens of canine distemper virus Onderstepoort and a recent field isolate, KDK-1, were investigated with strain specific dog sera. A conventional neutralization assay indicated antigenic dissimilarity between the strains; however, when guinea pig complement was included in the reaction mixture, the strains were neutralized with not only the homologous but also the heterologous antibodies. PMID- 12093698 TI - QuickVue influenza test for rapid detection of influenza A and B viruses in a pediatric population. AB - The performance of a lateral-flow immunoassay, the QuickVue Influenza Test, for detection of influenza A and B viruses in comparison with that of cell culture was evaluated by using nasopharyngeal aspirates, in viral transport medium, from children with respiratory tract infections. The sensitivity and specificity were 79.2 and 82.6%, respectively. PMID- 12093699 TI - Microimmunoassay using a protein chip: optimizing conditions for protein immobilization. AB - Optimizing conditions for the microarraying of protein antigens onto glass slides were studied. Various vendors, surface functional groups, buffers, and fixatives were evaluated to enhance protein binding. A total of 125 pg of human immunoglobulin was detectable with this assay system, suggesting that protein microarray can be applied for routine immunodiagnosis. PMID- 12093700 TI - Manipulations in maternal environment reverse periodontitis in genetically predisposed rats. AB - The predisposition to develop periodontitis is partly genetically determined in humans as well as in animals. Here we demonstrate, however, that early manipulations in the maternal environment of an animal (rat) model of periodontitis can fully reverse the genetic predisposition to develop periodontitis at adult age. PMID- 12093701 TI - Influence of antimalarial treatment on acquisition of immunity in Plasmodium berghei NK65 malaria. AB - Antimalarial treatments during primary Plasmodium berghei NK65 infection in BALB/c mice influenced the acquisition of protective immunity against reinfection. Among subcurative treatments, lower doses better enable mice to acquire protective immunity than do higher doses. Eradication of parasites from the start of infection did not promote protective immunity. PMID- 12093702 TI - Interleukin-8 and tumor necrosis factor alpha production in human epidermal keratinocytes induced by Trichophyton mentagrophytes. AB - Production of interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) was confirmed by enzyme-linked immunosorbent assay in a medium where human epidermal keratinocytes were cocultured with Trichophyton mentagrophytes for 1 to 12 h. IL-8 and TNF-alpha mRNAs were also detected in the keratinocytes cocultured with T. mentagrophytes. PMID- 12093703 TI - Widal test in diagnosis of typhoid fever in Turkey. AB - We studied the value of the Widal tube agglutination test for the diagnosis of typhoid fever. The subjects were all adults >18 years of age and were divided into four groups: (i) 317 healthy blood donor controls, (ii) 31 bacteriologically confirmed patients with Salmonella enterica serotype Typhi, (iii) 21 patients with a clinical diagnosis of typhoid fever, and (iv) 41 febrile nontyphoid patients. Blood donor controls were screened with a slide agglutination test for the Salmonella enterica serotype Typhi O and H antigens, and positives were then tested with the Widal test. Acute- and convalescent-phase sera from patients in groups 2, 3, and 4 were obtained 7 to 10 days apart and tested by the Widal test. Using a cutoff of >or = 1/200 for the O antigen test performed on acute-phase serum gave a sensitivity of 52% and a specificity of 88% with a positive predictive value (PPV) of 76% and a negative predictive value (NPV) of 71%. This increased to 90% sensitivity and specificity with a PPV of 88% and an NPV of 93% when the convalescent-phase serum was tested. We concluded that O and H agglutinin titers of > or = 1/200 are of diagnostic significance. The Widal test is easy, inexpensive, and relatively noninvasive. It can be of diagnostic value when blood cultures are not available or practical. The results must be interpreted cautiously because of the low sensitivity of the test. The Widal test done on convalescent-phase serum gave more-reliable results with higher specificity and sensitivity. PMID- 12093704 TI - False-positive human immunodeficiency virus seroconversion is not common following rabies vaccination. PMID- 12093705 TI - Heart rate variability in emergency department patients with sepsis. AB - OBJECTIVE: To test the hypothesis that heart rate variability (HRV) can provide an early indication of illness severity among patients presenting to the emergency department (ED) with sepsis. METHODS: The authors enrolled a convenience sample of 15 ED patients meeting the American College of Chest Physicians/Society of Critical Care Medicine criteria for sepsis. Each patient had continuous Holter monitoring performed in the ED. Acute Physiology and Chronic Health II (APACHE II) and Sequential Organ Failure (SOFA) scores were calculated for the day of presentation. Holter tapes obtained in the ED were analyzed off-line to calculate HRV variables for the 5-minute segment with the least artifact and non-sinus beats. These variables were correlated with APACHE II and SOFA scores. RESULTS: LFnu (normalized low-frequency power), an assessment of the relative sympathetic contribution to overall HRV, was correlated with increased illness severity as calculated using APACHE II (r = -0.67, r(2) = 0.43) and SOFA (r = -0.80, r(2) = 0.64) scores. LF/HF ratio (low-frequency/high frequency ratio), a measure of sympathovagal balance, was correlated with the SOFA score [r = -0.54 (95% CI = -0.83 to -0.01), r(2) = 0.29]. All five patients who required critical care monitoring or ventilatory support or who died during the first 5 days of their hospitalization had LFnu values below 0.5 and LF/HF ratios less than 1.0. None of the patients with measurements greater than these threshold values died or required these interventions during the five days following admission. CONCLUSIONS: A single variable, LFnu, which reflects sympathetic modulation of heart rate, accounted for 40-60% of the variance in illness severity scores among patients presenting to the ED with sepsis. HRV, as reflected in LFnu and the LF/HF ratio and measured with a single brief (5-minute) period of monitoring while in the ED, may provide the emergency physician with a readily available, noninvasive, early marker of illness severity. The threshold effect of LFnu and LF/HF in the prediction of early clinical deterioration was an unexpected finding and should be regarded as hypothesis-generating, pending further study. PMID- 12093706 TI - Derivation of a termination-of-resuscitation guideline for emergency medical technicians using automated external defibrillators. AB - OBJECTIVES: To determine the association between characteristics of cardiac arrest and survival to hospital discharge following failed resuscitation by defibrillation-trained emergency medical technicians (EMT-Ds), and to propose an out-of-hospital termination-of-resuscitation (TOR) guideline for EMT-Ds. METHODS: A 22-month retrospective review of 700 out-of-hospital primary cardiac arrest patients in a large emergency medical services (EMS) system who received exclusively EMT-D care. RESULTS: Seven hundred primary cardiac arrest patients were identified. Follow-up was obtained in 662 cases (94.6%). Of these, 36 (5.4%) achieved a return of spontaneous circulation (ROSC) prior to transport. Among the 626 patients who failed to achieve ROSC at any time, two (0.3%) survived to discharge. Multivariate analysis showed that ROSC at any time had the strongest association with survival [odds ratio (OR) 45.5; 95% confidence interval (95% CI) = 8.5 to 243.7]. A shock prior to transport (OR 6.9; 95% CI = 1.2 to 40.3) and cardiac arrest witnessed by EMS personnel (OR 4.4; 95% CI = 1.0 to 18.5) were also independently associated with survival. These variables were incorporated into a TOR guideline. The guideline was 100% sensitive (95% CI = 99.1 to 100) in identifying survivors and had 100% negative predictive value (95% CI = 75.3 to 100) for identifying nonsurvivors of out-of-hospital cardiac arrest in the study population. CONCLUSIONS: In this EMS system, cardiac arrest patients may be considered for out-of-hospital TOR following EMT-D resuscitation attempts when there has been no ROSC, no shock has been given, and the arrest was not witnessed by EMS personnel. These guidelines require prospective validation. PMID- 12093707 TI - Influence of international emergency medicine opportunities on residency program selection. AB - OBJECTIVES: In the last several years, there has been increasing interest in international emergency medicine (IEM) programs. A number of residency programs offer or encourage international opportunities during residency. The purpose of this study was twofold: 1) to determine whether the availability of international opportunities during residency affected the ranking of emergency medicine (EM) residency programs by graduating medical students and 2) to determine whether prior international health experience among medical students influences interviewing and ranking of residency programs with international opportunities. METHODS: An eight-question survey measuring interest in IEM and its effect on ranking of programs was mailed to all 1,205 first-year EM residents from 122 accredited allopathic EM residency programs in the United States. International opportunities were defined as any program that offered an international elective during residency or had an IEM fellowship. Data were analyzed using descriptive statistics and chi-square tests. RESULTS: Of the 1,205 mailed surveys, 34% were returned by the first-year residents. Of all respondents, 281/408 (69%) interviewed at programs with international opportunities. Study findings revealed 62% (164/264) of the students who interviewed at programs with international opportunities considered the availability of international opportunities a positive factor in ranking residency programs. Analysis revealed that applicants with previous international health experience, 68% (104/152), were more likely to rank EM residency programs with international opportunities higher than those that did not. CONCLUSIONS: This study suggests that the availability of international opportunities during residency positively affected residency ranking. This relationship was stronger in medical students with prior international health experience. PMID- 12093708 TI - Factors influencing admission among children with a traumatic brain injury. AB - OBJECTIVES: To describe the epidemiology of traumatic brain injury (TBI) among children in Maryland and to examine factors that influence hospital admission. METHODS: Statewide mortality, hospital discharge, and ambulatory care data were used to identify all TBI-related emergency department (ED) visits, hospitalizations, and deaths that occurred in 1998 to children aged 0-19 years according to the Centers for Disease Control and Prevention's standard case definition and protocol. Inpatient admission was modeled as a function of patient, injury, and hospital characteristics. RESULTS: The overall incidence of pediatric TBI (i.e., ED visits, hospitalizations, and deaths) in 1998 was 670/100,000. After controlling for injury severity and other factors, uninsured children were 40% less likely to be hospitalized (95% CI = 0.43 to 0.82) and children with Medicaid were 90% more likely to be hospitalized (95% CI = 1.42 to 2.54) than were those with private insurance. The presence of a major associated injury significantly influenced the likelihood of hospitalization, especially among children with a minor (OR = 8.8) to moderate (OR = 11.6) TBI. Children who presented to a trauma center hospital were significantly more likely to be hospitalized than children treated at a non-trauma center hospital, although this varied depending on income (OR = 1.8 for high versus low) and hospital volume (OR = 2.6 for a small hospital and OR = 29.0 for a large hospital). CONCLUSIONS: After adjusting for TBI severity and the presence of associated injuries, significant differences in hospitalization rates may exist among different patient subgroups and hospitals for children who sustain TBIs. PMID- 12093710 TI - Untreated hypercholesterolemia in an emergency department chest pain observation unit population. AB - OBJECTIVES: To describe the prevalence of hypercholesterolemia in a predominantly African American, innercity chest pain observation unit (CPOU) patient population, and to estimate the percentage of patients eligible for cholesterol lowering therapy as indicated by the 2001 National Cholesterol Education Program guidelines. METHODS: A cross-sectional study design utilizing a convenience sample of patients from a high-volume urban hospital CPOU. Patients with chest pain suspicious of cardiac etiology who had negative initial electrocardiograms and cardiac markers were assigned to the chest pain protocol. Consenting subjects were screened for hypercholesterolemia through capillary blood point-of-care testing with a cutoff of 190 mg/dL. Those who tested positive had four-hour fasting complete lipid profiles performed by the central laboratory. RESULTS: There were 112 patients enrolled in this study (mean age = 51 years; 57% male; and 83% African American). Elevated values on the screening test were obtained on 28 [25%; 95% confidence interval (95% CI) = 16.9 to 33.0] of these patients. These patients were found to have a mean four-hour fasting total cholesterol level of 224 mg/dL, a low-density lipoprotein (LDL) level of 138 mg/dL, a high density lipoprotein (HDL) level of 52 mg/dL, and a triglyceride level of 168 mg/dL. Of the patients identified through the screening test, 54% proved eligible for cholesterol-lowering medications and 91.7% of these patients reported an interest in initiating therapy. CONCLUSIONS: In this study, approximately 25% of inner-city CPOU patients are possible candidates for cholesterol-lowering interventions. Benefits of initiating therapy during this potential "teachable moment" in a CPOU should be investigated in a subsequent multicenter randomized trial. PMID- 12093709 TI - Injury mortality following the loss of air medical support for rural interhospital transport. AB - OBJECTIVES: This study evaluated variation in mortality among interfacility transfers three years before and after discontinuation of a rotor-wing transport service. METHODS: A retrospective cohort assessment was conducted among severely injured patients transferred from four rural hospitals to a single tertiary center in regions with continued versus discontinued rotor-wing service. Thirty day mortality following discharge from the receiving tertiary facility served as the primary outcome measure. RESULTS: Discontinuation of rotor-wing transport decreased interfacility transfers and increased transfer time. Transferred patients were four times more likely to die after (compared with before) rotor wing service was discontinued (p = 0.05). No difference was noted in the region with continued rotor-wing service [odds ratio (OR) = 0.53, p = 0.47]. CONCLUSIONS: Injury mortality increased with loss of air transport for interfacility transfer in a rural area. PMID- 12093711 TI - Clinicopathological conference: a simple case of abdominal pain. PMID- 12093712 TI - The paradox of gold standards. PMID- 12093713 TI - Fellowship training: a necessity in today's academic world. PMID- 12093714 TI - Whither the fourth link? PMID- 12093715 TI - Preventive care in the emergency department: diagnosis and management of smoking and smoking-related illness in the emergency department: a systematic review. AB - Smoking remains the leading preventable cause of morbidity and mortality in the United States. The efficacy of emergency department (ED)-based patient screening and counseling for smoking cessation is not currently known. OBJECTIVES: To perform a structured, systematic review of the medical literature to assess the efficacy of limited screening and counseling for tobacco use cessation among adults in the primary care and ED settings, and develop recommendations for emergency physicians based on these data. METHODS: A two-person template-driven review of all English-language articles from Medline, the Cochrane Database, and two recent smoking cessation guidelines, with evidence graded according to the scheme of the U.S. Preventive Services Task Force (USPSTF), was performed. Recommendations were developed, and strength graded, based on this evidence. RESULTS: Of 2,078 citations reviewed, 16 were selected for inclusion, based on their methodologic strength and relevance to emergency medicine. Routine physician screening and counseling may increase quit rates at 6-12 months from 3% (usual care) to 8-11%. Interventions include brief counseling (<3 minutes), possibly supplemented with self-help literature, nicotine replacement therapy (NRT), and follow-up telephone calls. CONCLUSIONS: Strong evidence exists, in the primary care setting, that smoking cessation screening and counseling are effective. Limited data exist for ED-based practice, but, based on the burden of disease, relative ease of intervention, and likely efficacy, routine screening of all patients for tobacco use and referral of smokers to primary care and cessation programs are recommended. PMID- 12093716 TI - Gamma hydroxybutyric acid (GHB) intoxication. AB - Gamma hydroxybutyric acid (GHB) is a naturally occurring analog of gamma aminobutyric acid (GABA) that has been used in research and clinical medicine for many years. In the past decade it has become very popular as a dietary supplement and recreational drug. Acute overdose leads to profound alteration of mental status and variable amounts of respiratory depression. With proper management, most patients recover fully within six hours. However, respiratory arrest and death have been reported in severe GHB intoxication. In addition to acute overdose, there is a GHB withdrawal syndrome that is similar to sedative/hypnotic and ethanol withdrawal. Recently several congeners of GHB, gamma butyrolactone and 1,4-butanediol, have emerged as drugs of abuse and show toxidromes similar to GHB. Emergency physicians should be familiar with the presentation and management of GHB-related emergencies. PMID- 12093717 TI - Diagnostic errors. AB - One of the most complex and challenging tasks facing physicians is the need to make a diagnosis. Recent research has focused attention on medical errors alleged to have resulted in increased patient morbidity and mortality. A number of authorities have suggested methods to track and prevent errors. Most have dealt with systems' changes and fail-safe methods to prevent medication errors and commission of errors during treatment. Few have addressed methods to find and prevent diagnostic errors. Unless diagnostic error is either avoided or corrected early, fail-safe methods to prevent medication and treatment errors will ultimately fail to improve patient outcome. American medical literature, particularly postmortem studies, have documented diagnostic error since at least 1912. European literature shows the problem is worldwide. The limits of human memory and errors in both observation and processing of information during problem solving contribute to the commission of errors. The purpose of this article is to examine the thinking patterns and cognitive errors that can result in diagnostic error, and suggest instructional strategies that can be used to alert residents and attending physicians to these potential problems so they can be avoided. PMID- 12093718 TI - Attitudes of law enforcement officers regarding automated external defibrillators. AB - OBJECTIVE: To assess the knowledge and attitudes of law enforcement officers regarding treating out-of-hospital cardiac arrest (OHCA) and using automated external defibrillators (AEDs). METHODS: This was a survey conducted among officers serving Marion County, Indiana. RESULTS: Of 1,130 surveys distributed, 929 (82.2%) were returned. Among these officers, 603 (66.4%) were certified in cardiopulmonary resuscitation (CPR) and 103 (11.3%) had received AED training. Most officers had limited knowledge regarding OHCA. A 100-point knowledge score (mean +/- SD: 31.9 +/- 14.3) was higher in officers who had performed CPR while on duty [35.0, 95% confidence interval (95% CI) = 32.9 to 37.2, p = 0.005] and who were AED-trained (40.8, 95% CI = 38.0 to 43.6, p < 0.001). Of the respondents, 367 (40.1%) believed that AED usage by local law enforcement was needed, and 323 (35.6%) stated that they would feel comfortable using an AED if trained. A 100-point attitude score (mean +/- SD: 32.1 +/- 21.0) was higher in officers who had CPR certification (38.2, 95% CI = 35.6 to 40.8), who had performed CPR while on duty (40.6, 95% CI = 37.7 to 43.5), who were AED-trained (39.5, 95% CI = 35.6 to 43.4), and who had improved OHCA knowledge (+3.8 per 10 points knowledge score, 95% CI = 3.0 to 4.7), p < 0.001, all significant factors. CONCLUSIONS: Limited knowledge and negative attitudes of law enforcement officers regarding their involvement in treating OHCA and using AEDs are commonly present. These factors could result in barriers that negatively impact law enforcement AED programs. PMID- 12093719 TI - Thrombolysis candidates for the treatment of stroke at an emergency department in Japan. AB - OBJECTIVES: To study the proportion and characteristics of potential candidates for the intravenous administration of tissue plasminogen activator (IV-tPA) among patients with cerebral infarction in a Japanese emergency department (ED). METHODS: A retrospective observational study was performed using patients who had been transported by ambulance between August 1988 and April 2000 to an urban ED of a university hospital located in the Tokyo metropolitan area. Potential candidates for IV-tPA were identified using the criteria of the National Institute of Neurological Disorders and Stroke (NINDS) study. RESULTS: Of all 30,064 patients transported by ambulance, 526 were diagnosed as having cerebral infarction. Among them, 190 patients arrived at the ED within two hours of symptom onset (early ED arrivers). In comparison of their demographics with late ED arrivers (n = 319), atrial fibrillation, male gender, and consciousness disturbance were related with early ED arrivers, while aging and diabetes were related with late ED arrivers. As to the stroke subtype, patients with an embolic infarction accounted for 76.8% among early ED arrivers. Application of exclusion criteria identified 114 patients, who were suitable for the thrombolysis treatment, indicating that the proportion of potential IV-tPA candidates was 21.7% among all cerebral infarction patients and 0.38% among all ED patients. CONCLUSIONS: The number of potential IV-tPA candidates among patients transported to the ED by ambulance in Japan was substantial, where the proportion of embolic infarction cases was extremely high. PMID- 12093720 TI - TCAs and myocardial function. PMID- 12093721 TI - Finding a solution for ambulance diversion. PMID- 12093722 TI - The "golden hour" paradigm. PMID- 12093725 TI - NO sensing by FNR: regulation of the Escherichia coli NO-detoxifying flavohaemoglobin, Hmp. AB - Nitric oxide (NO) is a signalling and defence molecule of major importance in biology. The flavohaemoglobin Hmp of Escherichia coli is involved in protective responses to NO. Because hmp gene transcription is repressed by the O(2) responsive regulator FNR, we investigated whether FNR also senses NO. The [4Fe 4S](2+) cluster of FNR is oxygen labile and controls protein dimerization and site-specific DNA binding. NO reacts anaerobically with the Fe-S cluster of purified FNR, generating spectral changes consistent with formation of a dinitrosyl-iron-cysteine complex. NO-inactivated FNR can be reconstituted, suggesting physiological relevance. FNR binds at an FNR box within the hmp promoter (P(hmp)). FNR samples inactivated by either O(2) or NO bind specifically to P(hmp), but with lower affinity. Dose-dependent up-regulation of P(hmp) in vivo by NO concentrations of pathophysiological relevance is abolished by fnr mutation, and NO also modulates expression from model FNR-regulated promoters. Thus, FNR can respond to not only O(2), but also NO, with major implications for global gene regulation in bacteria. We propose an NO-mediated mechanism of hmp regulation by which E.coli responds to NO challenge. PMID- 12093723 TI - Structure of coronavirus main proteinase reveals combination of a chymotrypsin fold with an extra alpha-helical domain. AB - The key enzyme in coronavirus polyprotein processing is the viral main proteinase, M(pro), a protein with extremely low sequence similarity to other viral and cellular proteinases. Here, the crystal structure of the 33.1 kDa transmissible gastroenteritis (corona)virus M(pro) is reported. The structure was refined to 1.96 A resolution and revealed three dimers in the asymmetric unit. The mutual arrangement of the protomers in each of the dimers suggests that M(pro) self-processing occurs in trans. The active site, comprised of Cys144 and His41, is part of a chymotrypsin-like fold that is connected by a 16 residue loop to an extra domain featuring a novel alpha-helical fold. Molecular modelling and mutagenesis data implicate the loop in substrate binding and elucidate S1 and S2 subsites suitable to accommodate the side chains of the P1 glutamine and P2 leucine residues of M(pro) substrates. Interactions involving the N-terminus and the alpha-helical domain stabilize the loop in the orientation required for trans cleavage activity. The study illustrates that RNA viruses have evolved unprecedented variations of the classical chymotrypsin fold. PMID- 12093724 TI - Crystal structure of the middle and C-terminal domains of the flagellar rotor protein FliG. AB - The FliG protein is essential for assembly, rotation and clockwise/counter clockwise (CW/CCW) switching of the bacterial flagellum. About 25 copies of FliG are present in a large rotor-mounted assembly termed the 'switch complex', which also contains the proteins FliM and FliN. Mutational studies have identified the segments of FliG most crucial for flagellar assembly, rotation and switching. The structure of the C-terminal domain, which functions specifically in rotation, was reported previously. Here, we describe the crystal structure of a larger fragment of the FliG protein from Thermotoga maritima, which encompasses the middle and C terminal parts of the protein (termed FliG-MC). The FliG-MC molecule consists of two compact globular domains, linked by an alpha-helix and an extended segment that contains a well-conserved Gly-Gly motif. Mutational studies indicate that FliM binds to both of the globular domains, and given the flexibility of the linking segment, FliM is likely to determine the relative orientation of the domains in the flagellum. We propose a model for the organization of FliG-MC molecules in the flagellum, and suggest that CW/CCW switching might occur by movement of the C-terminal domain relative to other parts of FliG, under the control of FliM. PMID- 12093726 TI - Structural analysis of two enzymes catalysing reverse metabolic reactions implies common ancestry. AB - The crystal structure of the dimeric anthranilate phosphoribosyltransferase (AnPRT) reveals a new category of phosphoribosyltransferases, designated as class III. The active site of this enzyme is located within the flexible hinge region of its two-domain structure. The pyrophosphate moiety of phosphoribosylpyrophosphate is co-ordinated by a metal ion and is bound by two conserved loop regions within this hinge region. With the structure of AnPRT available, structural analysis of all enzymatic activities of the tryptophan biosynthesis pathway is complete, thereby connecting the evolution of its enzyme members to the general development of metabolic processes. Its structure reveals it to have the same fold, topology, active site location and type of association as class II nucleoside phosphorylases. At the level of sequences, this relationship is mirrored by 13 structurally invariant residues common to both enzyme families. Taken together, these data imply common ancestry of enzymes catalysing reverse biological processes--the ribosylation and deribosylation of metabolic pathway intermediates. These relationships establish new links for enzymes involved in nucleotide and amino acid metabolism. PMID- 12093728 TI - FXYD7 is a brain-specific regulator of Na,K-ATPase alpha 1-beta isozymes. AB - Recently, corticosteroid hormone-induced factor (CHIF) and the gamma-subunit, two members of the FXYD family of small proteins, have been identified as regulators of renal Na,K-ATPase. In this study, we have investigated the tissue distribution and the structural and functional properties of FXYD7, another family member which has not yet been characterized. Expressed exclusively in the brain, FXYD7 is a type I membrane protein bearing N-terminal, post-translationally added modifications on threonine residues, most probably O-glycosylations that are important for protein stabilization. Expressed in Xenopus oocytes, FXYD7 can interact with Na,K-ATPase alpha 1-beta 1, alpha 2-beta 1 and alpha 3-beta 1 but not with alpha-beta 2 isozymes, whereas, in brain, it is only associated with alpha 1-beta isozymes. FXYD7 decreases the apparent K(+) affinity of alpha 1-beta 1 and alpha 2-beta 1, but not of alpha 3-beta1 isozymes. These data suggest that FXYD7 is a novel, tissue- and isoform-specific Na,K-ATPase regulator which could play an important role in neuronal excitability. PMID- 12093727 TI - Cytoplasmic transport of Stat3 by receptor-mediated endocytosis. AB - Signal transducer and activator of transcription (STAT) proteins are cytoplasmic transcription factors that translocate to the nucleus and regulate gene expression upon activation of cytokine or growth factor receptors. While this translocation event is essential for gene regulation by STATs, their mechanism of transport through the cytoplasm to the nucleus has remained elusive. We now report that cytoplasmic transport of Stat3 is an active process that requires receptor-mediated endocytosis. Stat3 co-localizes with endocytic vesicles in transit from the cell membrane to the perinuclear region in response to growth factor stimulation. Consistent with a role for receptor endocytosis in growth factor signaling, disruption of endocytosis with specific inhibitors blocks Stat3 nuclear translocation and Stat3-dependent gene regulation. These results indicate that receptor-mediated endocytosis may be a general mechanism of transport through the cytoplasm for a subset of cytoplasmic signaling proteins destined for the nucleus. PMID- 12093729 TI - Involvement of Fes/Fps tyrosine kinase in semaphorin3A signaling. AB - Collapsin response mediator proteins (CRMPs)/TOAD64/Ulips/DRPs and CRAM have emerged as strong candidates for a role in semaphorin signaling. In this study we identified Fes/Fps (Fes) tyrosine kinase in the CRMP-CRAM complex and investigated whether Fes was involved in semaphorin3A (Sema3A) signaling. In COS 7 cells, the interaction between Fes and plexinA1 (PlexA1) and the tyrosine phosphorylation of PlexA1 by Fes were observed; however, these events were significantly attenuated by co-expression of neuropilin-1 (NP-1). Even with NP-1 co-expression, Sema3A was able to enhance the association of Fes with PlexA1 and Fes-mediated tyrosine phosphorylation of PlexA1, CRAM and CRMP2. Co-expression of Fes with PlexA1 exhibited COS-7 cell contraction activity, indicating that Fes can convert inactive PlexA1 to its active form, whereas combination of Fes/NP 1/PlexA1 or Fes kinase-negative mutants/PlexA1 did not alter cell morphology. Finally, Sema3A-induced growth cone collapse of dorsal root ganglion neurons was suppressed by expression of Fes kinase-negative mutants. Taken together, our findings suggest that Fes links Sema3A signals to CRMP-CRAM, and that NP-1 negatively regulates PlexA1 activation by Fes in resting condition. PMID- 12093730 TI - Structural basis for the reversible activation of a Rho protein by the bacterial toxin SopE. AB - The bacterial enteropathogen Salmonella typhimurium employs a type III secretion system to inject bacterial toxins into the host cell cytosol. These toxins transiently activate Rho family GTP-binding protein-dependent signaling cascades to induce cytoskeletal rearrangements. One of these translocated Salmonella toxins, SopE, can activate Cdc42 in a Dbl-like fashion despite its lack of sequence similarity to Dbl-like proteins, the Rho-specific eukaryotic guanine nucleotide exchange factors. To elucidate the mechanism of SopE-mediated guanine nucleotide exchange, we have analyzed the structure of the complex between a catalytic fragment of SopE and Cdc42. SopE binds to and locks the switch I and switch II regions of Cdc42 in a conformation that promotes guanine nucleotide release. This conformation is strikingly similar to that of Rac1 in complex with the eukaryotic Dbl-like exchange factor Tiam1. However, the catalytic domain of SopE has an entirely different architecture from that of Tiam1 and interacts with the switch regions via different amino acids. Therefore, SopE represents the first example of a non-Dbl-like protein capable of inducing guanine nucleotide exchange in Rho family proteins. PMID- 12093731 TI - Involvement of the mitogen-activated protein kinase SIMK in regulation of root hair tip growth. AB - Mitogen-activated protein kinases (MAPKs) are involved in stress signaling to the actin cytoskeleton in yeast and animals. We have analyzed the function of the stress-activated alfalfa MAP kinase SIMK in root hairs. In epidermal cells, SIMK is predominantly nuclear. During root hair formation, SIMK was activated and redistributed from the nucleus into growing tips of root hairs possessing dense F actin meshworks. Actin depolymerization by latrunculin B resulted in SIMK relocation to the nucleus. Conversely, upon actin stabilization with jasplakinolide, SIMK co-localized with thick actin cables in the cytoplasm. Importantly, latrunculin B and jasplakinolide were both found to activate SIMK in a root-derived cell culture. Loss of tip-focused SIMK and actin was induced by the MAPK kinase inhibitor UO 126 and resulted in aberrant root hairs. UO 126 inhibited targeted vesicle trafficking and polarized growth of root hairs. In contrast, overexpression of gain-of-function SIMK induced rapid tip growth of root hairs and could bypass growth inhibition by UO 126. These data indicate that SIMK plays a crucial role in root hair tip growth. PMID- 12093732 TI - Stress-inducible protein 1 is a cell surface ligand for cellular prion that triggers neuroprotection. AB - Prions are composed of an isoform of a normal sialoglycoprotein called PrP(c), whose physiological role has been under investigation, with focus on the screening for ligands. Our group described a membrane 66 kDa PrP(c)-binding protein with the aid of antibodies against a peptide deduced by complementary hydropathy. Using these antibodies in western blots from two-dimensional protein gels followed by sequencing the specific spot, we have now identified the molecule as stress-inducible protein 1 (STI1). We show that this protein is also found at the cell membrane besides the cytoplasm. Both proteins interact in a specific and high affinity manner with a K(d) of 10(-7) M. The interaction sites were mapped to amino acids 113-128 from PrP(c) and 230-245 from STI1. Cell surface binding and pull-down experiments showed that recombinant PrP(c) binds to cellular STI1, and co-immunoprecipitation assays strongly suggest that both proteins are associated in vivo. Moreover, PrP(c) interaction with either STI1 or with the peptide we found that represents the binding domain in STI1 induce neuroprotective signals that rescue cells from apoptosis. PMID- 12093734 TI - GH3, a novel proapoptotic domain in Drosophila Grim, promotes a mitochondrial death pathway. AB - Grim encodes a protein required for programmed cell death in Drosophila. The Grim N-terminus induces apoptosis by disrupting IAP blockage of caspases; however, N terminally-deleted Grim retains pro apoptotic activity. We describe GH3, a 15 amino acid internal Grim domain absolutely required for its proapoptotic activity and sufficient to induce cell death when fused to heterologous carrier proteins. A GH3 homology region is present in the Drosophila proapoptotic proteins Reaper and Sickle. The GH3 domain and the homologous regions in Reaper and Sickle are predicted to be structured as amphipathic alpha-helixes. During apoptosis induction, Grim colocalizes with mitochondria and cytochrome c in a GH3-dependent but N-terminal- and caspase activity-independent manner. When Grim is overexpressed in vivo, both the N-terminal and the GH3 domains are equally necessary, and cooperate for apoptosis induction. The N-terminal and GH3 Grim domains thus activate independent apoptotic pathways that synergize to induce programmed cell death efficiently. PMID- 12093735 TI - Telencephalon-specific Rb knockouts reveal enhanced neurogenesis, survival and abnormal cortical development. AB - Correct cell cycle regulation and terminal mitosis are critical for nervous system development. The retinoblastoma (Rb) protein is a key regulator of these processes, as Rb-/- embryos die by E15.5, exhibiting gross hematopoietic and neurological defects. The extensive apoptosis in Rb-/- embryos has been attributed to aberrant S phase entry resulting in conflicting growth control signals in differentiating cells. To assess the role of Rb in cortical development in the absence of other embryonic defects, we examined mice with telencephalon-specific Rb deletions. Animals carrying a floxed Rb allele were interbred with mice in which cre was knocked into the Foxg1 locus. Unlike germline knockouts, mice specifically deleted for Rb in the developing telencephalon survived until birth. In these mutants, Rb-/- progenitor cells divided ectopically, but were able to survive and differentiate. Mutant brains exhibited enhanced cellularity due to increased proliferation of neuroblasts. These studies demonstrate that: (i) cell cycle deregulation during differentiation does not necessitate apoptosis; (ii) Rb-deficient mutants exhibit enhanced neuroblast proliferation; and (iii) terminal mitosis may not be required to initiate differentiation. PMID- 12093733 TI - Cellular prion protein transduces neuroprotective signals. AB - To test for a role for the cellular prion protein (PrP(c)) in cell death, we used a PrP(c)-binding peptide. Retinal explants from neonatal rats or mice were kept in vitro for 24 h, and anisomycin (ANI) was used to induce apoptosis. The peptide activated both cAMP/protein kinase A (PKA) and Erk pathways, and partially prevented cell death induced by ANI in explants from wild-type rodents, but not from PrP(c)-null mice. Neuroprotection was abolished by treatment with phosphatidylinositol-specific phospholipase C, with human peptide 106-126, with certain antibodies to PrP(c) or with a PKA inhibitor, but not with a MEK/Erk inhibitor. In contrast, antibodies to PrP(c) that increased cAMP also induced neuroprotection. Thus, engagement of PrP(c) transduces neuroprotective signals through a cAMP/PKA-dependent pathway. PrP(c) may function as a trophic receptor, the activation of which leads to a neuroprotective state. PMID- 12093736 TI - Division site placement in E.coli: mutations that prevent formation of the MinE ring lead to loss of the normal midcell arrest of growth of polar MinD membrane domains. AB - The MinE protein functions as a topological specificity factor in determining the site of septal placement in Escherichia coli. MinE assembles into a membrane associated ring structure near midcell and directs the localization of MinD and MinC into a membrane- associated polar zone that undergoes a characteristic pole to-pole oscillation cycle. Single (green fluorescent protein) and double label (yellow fluorescent protein/cyan fluorescent protein) fluorescence labeling experiments showed that mutational alteration of a site on the alpha-face of MinE led to a failure to assemble the MinE ring, associated with loss of the ability to support a normal pattern of division site placement. The absence of the MinE ring did not prevent the assembly and disassembly of the MinD polar zone. Mutant cells lacking the MinE ring were characterized by the growth of MinD polar zones past their normal arrest point near midcell. The results suggested that the MinE ring acts as a stop-growth mechanism to prevent the MinCD polar zone from extending beyond the midcell division site. PMID- 12093737 TI - Deconstructing PML-induced premature senescence. AB - In this study, we investigated the subcellular and molecular mechanisms underlying promyelocytic leukemia (PML)-induced premature senescence. We demonstrate that intact PML nuclear bodies are not required for the induction of senescence. We have determined further that of seven known PML isoforms, only PML IV is capable of causing premature senescence, providing the first evidence for functional differences among these isoforms. Of interest is the fact that in contrast to PML(+/+) fibroblasts, PML(-/-) cells are resistant to PML IV-induced senescence. This suggests that although PML IV is necessary for this process to occur, it is not sufficient and requires other components for activity. Finally, we provide evidence that PML IV-induced senescence involves stabilization and activation of p53 through phosphorylation at Ser46 and acetylation at Lys382, and that it occurs independently of telomerase and differs from that elicited by oncogenic Ras. Taken together, our data assign a specific pro-senescent activity to an individual PML isoform that involves p53 activation and is independent from PML nuclear bodies. PMID- 12093738 TI - G(1)/S CDK is inhibited to restrain mitotic onset when DNA replication is blocked in fission yeast. AB - Cyclin-dependent kinase (CDK) Tyr15 phosphorylation plays a major role in regulating G(2)/M CDKs, but the role of this phosphorylation in regulating G(1)/S CDKs is less clear. We have studied the regulation and function of Cdc2-Tyr15 phosphorylation in the fission yeast Schizosaccharomyces pombe G(1)/S CDK Cig2/Cdc2. This complex is subject to high level Cdc2-Tyr15 phosphorylation inhibiting its kinase activity in hydroxyurea-treated cells blocked in S-phase. We show that this Tyr15 phosphorylation is required to maintain efficient mitotic checkpoint arrest, because Cig2 accumulates during the block and this accumulation can advance mitotic onset. This mitotic induction operates, at least in part, through activation of the normal G(2)/M CDK complex Cdc13/Cdc2. Thus, Tyr15 phosphorylation of G(1)/S CDK complexes is important in the checkpoint control blocking mitotic onset when DNA replication is inhibited. PMID- 12093739 TI - Drosophila cyclin E interacts with components of the Brahma complex. AB - Cyclin E-Cdk2 is essential for S phase entry. To identify genes interacting with cyclin E, we carried out a genetic screen using a hypomorphic mutation of Drosophila cyclin E (DmcycE(JP)), which gives rise to adults with a rough eye phenotype. Amongst the dominant suppressors of DmcycE(JP), we identified brahma (brm) and moira (mor), which encode conserved core components of the Drosophila Brm complex that is highly related to the SWI-SNF ATP-dependent chromatin remodeling complex. Mutations in genes encoding other Brm complex components, including snr1 (BAP45), osa and deficiencies that remove BAP60 and BAP111 can also suppress the DmcycE(JP) eye phenotype. We show that Brm complex mutants suppress the DmcycE(JP) phenotype by increasing S phases without affecting DmcycE protein levels and that DmcycE physically interacts with Brm and Snr1 in vivo. These data suggest that the Brm complex inhibits S phase entry by acting downstream of DmcycE protein accumulation. The Brm complex also physically interacts weakly with Drosophila retinoblastoma (Rbf1), but no genetic interactions were detected, suggesting that the Brm complex and Rbf1 act largely independently to mediate G(1) arrest. PMID- 12093740 TI - A growth factor-dependent nuclear kinase phosphorylates p27(Kip1) and regulates cell cycle progression. AB - The cyclin-dependent kinase inhibitor, p27(Kip1), which regulates cell cycle progression, is controlled by its subcellular localization and subsequent degradation. p27(Kip1) is phosphorylated on serine 10 (S10) and threonine 187 (T187). Although the role of T187 and its phosphorylation by Cdks is well-known, the kinase that phosphorylates S10 and its effect on cell proliferation has not been defined. Here, we identify the kinase responsible for S10 phosphorylation as human kinase interacting stathmin (hKIS) and show that it regulates cell cycle progression. hKIS is a nuclear protein that binds the C-terminal domain of p27(Kip1) and phosphorylates it on S10 in vitro and in vivo, promoting its nuclear export to the cytoplasm. hKIS is activated by mitogens during G(0)/G(1), and expression of hKIS overcomes growth arrest induced by p27(Kip1). Depletion of KIS using small interfering RNA (siRNA) inhibits S10 phosphorylation and enhances growth arrest. p27(-/-) cells treated with KIS siRNA grow and progress to S/G(2 )similar to control treated cells, implicating p27(Kip1) as the critical target for KIS. Through phosphorylation of p27(Kip1) on S10, hKIS regulates cell cycle progression in response to mitogens. PMID- 12093741 TI - Physiological and retinoid-induced proliferations of epidermis basal keratinocytes are differently controlled. AB - To investigate the roles of retinoic acid (RA) receptors (RARs) in the physiology of epidermis that does not express RAR beta, conditional spatio-temporally controlled somatic mutagenesis was used to selectively ablate RAR alpha in keratinocytes of RAR gamma-null mice. Keratinocyte proliferation was maintained in adult mouse epidermis lacking both RAR alpha and RAR gamma, as well as in RAR beta-null mice. All RAR-mediated signalling pathways are therefore dispensable in epidermis for homeostatic keratinocyte renewal. However, topical treatment of mouse skin with selective retinoids indicated that RXR/RAR gamma heterodimers, in which RXR transcriptional activity was subordinated to that of its RAR gamma partner, were required for retinoid-induced epidermal hyperplasia, whereas RXR homodimers and RXR/RAR alpha heterodimers were not involved. RA-induced keratinocyte proliferation was studied in mutant mice in which RXR alpha, RXR alpha and RAR alpha, RAR gamma, or RXR alpha and RAR gamma genes were specifically disrupted in either basal or suprabasal keratinocytes. We demonstrate that the topical retinoid signal is transduced by RXR alpha/RAR gamma heterodimers in suprabasal keratinocytes, which, in turn, stimulate proliferation of basal keratinocytes via a paracrine signal that may be heparin-binding EGF like growth factor. PMID- 12093742 TI - FANCE: the link between Fanconi anaemia complex assembly and activity. AB - The Fanconi anaemia (FA) nuclear complex (composed of the FA proteins A, C, G and F) is essential for protection against chromosome breakage. It activates the downstream protein FANCD2 by monoubiquitylation; this then forges an association with the BRCA1 protein at sites of DNA damage. Here we show that the recently identified FANCE protein is part of this nuclear complex, binding both FANCC and FANCD2. Indeed, FANCE is required for the nuclear accumulation of FANCC and provides a critical bridge between the FA complex and FANCD2. Disease-associated FANCC mutants do not bind to FANCE, cannot accumulate in the nucleus and are unable to prevent chromosome breakage. PMID- 12093743 TI - Mapping histone fold TAFs within yeast TFIID. AB - The transcription factor TFIID is a large multiprotein complex, composed of the TATA box-binding protein (TBP) and 14 TBP-associated factors (TAFs), which plays a key role in the regulation of gene expression by RNA polymerase II. The three dimensional structure of yeast (y) TFIID, determined at approximately 3 nm resolution by electron microscopy and image analysis, resembles a molecular clamp formed by three major lobes connected by thin linking domains. The yTFIID is structurally similar to the human factor although the clamp appears more closed in the yeast complex, probably reflecting the conformational flexibility of the structure. Immunolabelling experiments showed that nine TAFs that contain the histone fold structural motif were located in three distinct substructures of TFIID. The distribution of these TAFs showed that the previously reported pair wise interactions between histone fold domain (HFD)-containing TAFs are likely to occur in the native yTFIID complex. Most of the HFD-containing TAFs have been found in two distinct lobes, thus revealing an unexpected and novel molecular organization of TFIID. PMID- 12093744 TI - An embryonic/fetal beta-type globin gene repressor contains a nuclear receptor TR2/TR4 heterodimer. AB - We recently described an erythroid epsilon-globin gene repressor activity, which we named DRED (direct repeat erythroid-definitive). We show that DRED binds with high affinity to DR1 sites in the human embryonic (epsilon-) and fetal (gamma-) globin gene promoters, but the adult beta-globin promoter has no DR1 element. DRED is a 540 kDa complex; sequence determination showed that it contains the nuclear orphan receptors TR2 and TR4. TR2 and TR4 form a heterodimer that binds to the epsilon and gamma promoter DR1 sites. One mutation in a DR1 site causes elevated gamma-globin transcription in human HPFH (hereditary persistence of fetal hemoglobin) syndrome, and we show that this mutation reduces TR2/TR4 binding in vitro. The two receptor mRNAs are expressed at all stages of murine and human erythropoiesis; their forced transgenic expression reduces endogenous embryonic epsilony-globin transcription. These data suggest that TR2/TR4 forms the core of a larger DRED complex that represses embryonic and fetal globin transcription in definitive erythroid cells, and therefore that inhibition of its activity might be an attractive intervention point for treating sickle cell anemia. PMID- 12093745 TI - Formation of an hER alpha-COUP-TFI complex enhances hER alpha AF-1 through Ser118 phosphorylation by MAPK. AB - The enhancement of the human estrogen receptor alpha (hER alpha, NR3A1) activity by the orphan nuclear receptor COUP-TFI is found to depend on the establishment of a tight hER alpha-COUP-TFI complex. Formation of this complex seems to involve dynamic mechanisms different from those allowing hER alpha homodimerization. Although the hER alpha-COUP-TFI complex is present in all cells tested, the transcriptional cooperation between the two nuclear receptors is restricted to cell lines permissive to hER alpha activation function 1 (AF-1). In these cells, the physical interaction between COUP-TFI and hER alpha increases the affinity of hER alpha for ERK2/p42(MAPK), resulting in an enhanced phosphorylation state of the hER alpha Ser118. hER alpha thus acquires a strengthened AF-1 activity due to its hyperphosphorylation. These data indicate an alternative interaction process between nuclear receptors and demonstrate a novel protein intercommunication pathway that modulates hER alpha AF-1. PMID- 12093746 TI - The Runx3 transcription factor regulates development and survival of TrkC dorsal root ganglia neurons. AB - The RUNX transcription factors are important regulators of linage-specific gene expression in major developmental pathways. Recently, we demonstrated that Runx3 is highly expressed in developing cranial and dorsal root ganglia (DRGs). Here we report that within the DRGs, Runx3 is specifically expressed in a subset of neurons, the tyrosine kinase receptor C (TrkC) proprioceptive neurons. We show that Runx3-deficient mice develop severe limb ataxia due to disruption of monosynaptic connectivity between intra spinal afferents and motoneurons. We demonstrate that the underlying cause of the defect is a loss of DRG proprioceptive neurons, reflected by a decreased number of TrkC-, parvalbumin- and beta-galactosidase-positive cells. Thus, Runx3 is a neurogenic TrkC neuron specific transcription factor. In its absence, TrkC neurons in the DRG do not survive long enough to extend their axons toward target cells, resulting in lack of connectivity and ataxia. The data provide new genetic insights into the neurogenesis of DRGs and may help elucidate the molecular mechanisms underlying somatosensory-related ataxia in humans. PMID- 12093747 TI - The TRAP100 component of the TRAP/Mediator complex is essential in broad transcriptional events and development. AB - The multisubunit TRAP/Mediator complex is a mammalian counterpart of the yeast Mediator that shows diverse coactivation functions. Genetic ablation of the murine TRAP100 component of this complex has revealed that it is not essential for cell viability per se. However, null mutant mice die at an early developmental stage with severe malformations, and cultured TRAP100-deficient cells exhibit attenuated functions of a wide variety of transcriptional activators on ectopic reporters. The TRAP100-deficient TRAP/Mediator complex also lacks TRAP95 and TRAP150 beta/SUR2, which together with TRAP100 may form a submodule, and contains a reduced amount of SRB10/CDK8. Nevertheless, the residual complex shows unaltered binding both to RNA polymerase II and, with the exception of the oncoprotein E1A, to various activators. These findings suggest that TRAP/Mediator is broadly involved in transcription and that a TRAP100 containing submodule plays a secondary role, beyond primary activator interactions and RNA polymerase recruitment by the TRAP complex, in magnifying effects of activators on the general transcriptional machinery. PMID- 12093748 TI - Molecular basis of sequence-specific single-stranded DNA recognition by KH domains: solution structure of a complex between hnRNP K KH3 and single-stranded DNA. AB - To elucidate the basis of sequence-specific single-stranded (ss) DNA recognition by K homology (KH) domains, we have solved the solution structure of a complex between the KH3 domain of the transcriptional regulator heterogeneous nuclear ribonucleoprotein K (hnRNP K) and a 10mer ssDNA. We show that hnRNP K KH3 specifically recognizes a tetrad of sequence 5'd-TCCC. The complex is stabilized by a dense network of methyl-oxygen hydrogen bonds involving the methyl groups of three isoleucine residues and the O2 and N3 atoms of the two central cytosine bases. Comparison with the recently solved structure of a specific protein-ssDNA complex involving the KH3 and KH4 domains of the far upstream element (FUSE) binding protein FBP suggests that the amino acid located five residues N-terminal of the invariant GXXG motif, which is characteristic of all KH domains, plays a crucial role in discrimination of the first two bases of the tetrad. PMID- 12093749 TI - Novel nuclear and mitochondrial glycosylases revealed by disruption of the mouse Nth1 gene encoding an endonuclease III homolog for repair of thymine glycols. AB - Endonuclease III, encoded by nth in Escherichia coli, removes thymine glycols (Tg), a toxic oxidative DNA lesion. To determine the biological significance of this repair in mammals, we established a mouse model with mutated mNth1, a homolog of nth, by gene targeting. The homozygous mNth1 mutant mice showed no detectable phenotypical abnormality. Embryonic cells with or without wild-type mNth1 showed no difference in sensitivity to menadione or hydrogen peroxide. Tg produced in the mutant mouse liver DNA by X-ray irradiation disappeared with time, though more slowly than in the wild-type mouse. In extracts from mutant mouse liver, we found, instead of mNTH1 activity, at least two novel DNA glycosylase activities against Tg. One activity is significantly higher in the mutant than in wild-type mouse in mitochondria, while the other is another nuclear glycosylase for Tg. These results underscore the importance of base excision repair of Tg both in the nuclei and mitochondria in mammals. PMID- 12093750 TI - Formation of a nucleoprotein complex containing Tn7 and its target DNA regulates transposition initiation. AB - Tn7 insertion into its specific target site, attTn7, is mediated by the proteins TnsA, TnsB, TnsC and TnsD. The double-strand breaks that separate Tn7 from the donor DNA require the Tns proteins, the transposon and an attTn7 target DNA, suggesting that a prerequisite for transposition is the formation of a nucleoprotein complex containing TnsABC+D, and these DNAs. Here, we identify a TnsABC+D transposon-attTn7 complex, and demonstrate that it is a transposition intermediate. We demonstrate that an interaction between TnsB, the transposase subunit that binds to the transposon ends, and TnsC, the target DNA-binding protein that controls the activity of the transposase, is essential for assembly of the TnsABC+D transposon-attTn7 complex. We also show that certain TnsB residues are required for recombination because they mediate a TnsB-TnsC interaction critical to formation of the TnsABC+D transposon-attTn7 complex. We demonstrate that TnsA, the other transposase subunit, which also interacts with TnsC, greatly stabilizes the TnsABC+D transposon-attTn7 complex. Thus multiple interactions between the transposase subunits, TnsA and TnsB, and the target binding transposase activator, TnsC, control Tn7 transposition. PMID- 12093751 TI - Metal ions bound at the active site of the junction-resolving enzyme T7 endonuclease I. AB - T7 endonuclease I is a nuclease that is selective for the structure of the four way DNA junction. The active site is similar to those of a number of restriction enzymes. We have solved the crystal structure of endonuclease I with a wild-type active site. Diffusion of manganese ions into the crystal revealed two peaks of electron density per active site, defining two metal ion-binding sites. Site 1 is fully occupied, and the manganese ion is coordinated by the carboxylate groups of Asp55 and Glu65, and the main chain carbonyl of Thr66. Site 2 is partially occupied, and the metal ion has a single protein ligand, the remaining carboxylate oxygen atom of Asp55. Isothermal titration calorimetry showed the sequential exothermic binding of two manganese ions in solution, with dissociation constants of 0.58 +/- 0.019 and 14 +/- 1.5 mM. These results are consistent with a two metal ion mechanism for the cleavage reaction, in which the hydrolytic water molecule is contained in the first coordination sphere of the site 1-bound metal ion. PMID- 12093752 TI - PA200, a nuclear proteasome activator involved in DNA repair. AB - We have identified a novel 200 kDa nuclear protein that activates the proteasome. The protein, which we call PA200, has been purified to homogeneity from bovine testis and has been shown to activate proteasomal hydrolysis of peptides, but not proteins. Following gamma-irradiation of HeLa cells the uniform nuclear distribution of PA200 changes to a strikingly punctate pattern, a behavior characteristic of many DNA repair proteins. Homologs of PA200 are present in worms, plants and yeast. Others have shown that mutation of yeast PA200 results in hypersensitivity to bleomycin, and exposure of yeast to DNA damaging agents induces the PA200 message. Taken together, these findings implicate PA200 in DNA repair, possibly by recruiting proteasomes to double strand breaks. PMID- 12093753 TI - The yeast THO complex and mRNA export factors link RNA metabolism with transcription and genome instability. AB - The THO complex is a multimeric factor containing four polypeptides, Tho2, Hpr1, Mft1 and Thp2. Mutations in any of the genes encoding THO confer impairment of transcription and a transcription-dependent hyper-recombination phenotype, suggesting that THO has a functional role in gene expression. Using an in vivo assay developed to study expression of long and G+C-rich DNA sequences, we have isolated SUB2, a gene involved in mRNA splicing and export, as a multicopy suppressor of the gene expression defect of hpr1 Delta. Further investigation of a putative functional relationship between mRNA metabolism and THO revealed that mRNA export mutants sub2, yra1, mex67 and mtr2 have similar defective transcription and hyper-recombination phenotypes as THO mutants. In addition, THO becomes essential in cells with a defective Mex67 mRNA export er. Finally, we have shown that THO has the ability to associate with RNA and DNA in vitro. These results indicate a functional link between the processes of elongation and metabolism of nascent mRNA mediated by THO and mRNA export proteins, which have important consequences for the maintenance of genome stability. PMID- 12093755 TI - Structures of the pleiotropic translational regulator Hfq and an Hfq-RNA complex: a bacterial Sm-like protein. AB - In prokaryotes, Hfq regulates translation by modulating the structure of numerous RNA molecules by binding preferentially to A/U-rich sequences. To elucidate the mechanisms of target recognition and translation regulation by Hfq, we determined the crystal structures of the Staphylococcus aureus Hfq and an Hfq-RNA complex to 1.55 and 2.71 A resolution, respectively. The structures reveal that Hfq possesses the Sm-fold previously observed only in eukaryotes and archaea. However, unlike these heptameric Sm proteins, Hfq forms a homo-hexameric ring. The Hfq-RNA structure reveals that the single-stranded hepta-oligoribonucleotide binds in a circular conformation around a central basic cleft, whereby Tyr42 residues from adjacent subunits stack with six of the bases, and Gln8, outside the Sm motif, provides key protein-base contacts. Such binding suggests a mechanism for Hfq function. PMID- 12093754 TI - The exon junction complex is detected on CBP80-bound but not eIF4E-bound mRNA in mammalian cells: dynamics of mRNP remodeling. AB - Newly spliced mRNAs in mammalian cells are characterized by a complex of proteins at exon-exon junctions. This complex recruits Upf3 and Upf2, which function in nonsense-mediated mRNA decay (NMD). Both Upf proteins are detected on mRNA bound by the major nuclear cap-binding proteins CBP80/CBP20 but not mRNA bound by the major cytoplasmic cap-binding protein eIF4E. These and other data indicate that NMD targets CBP80-bound mRNA during a 'pioneer' round of translation, but whether nuclear eIF4E also binds nascent but dead-end transcripts is unclear. Here we provide evidence that nuclear CBP80 but not nuclear eIF4E is readily detected in association with intron-containing RNA and the C-terminal domain of RNA polymerase II. Consistent with this evidence, we demonstrate that RNPS1, Y14, SRm160, REF/Aly, TAP, Upf3X and Upf2 are detected in the nuclear fraction on CBP80-bound but not eIF4E-bound mRNA. Each of these proteins is also detected on CBP80-bound mRNA in the cytoplasmic fraction, indicating a presence on mRNA after export. The dynamics of mRNP composition before and after mRNA export are discussed. PMID- 12093758 TI - Cardiac chimerism as a mechanism for self-repair: does it happen and if so to what degree? PMID- 12093756 TI - Cryo-EM reveals an active role for aminoacyl-tRNA in the accommodation process. AB - During the elongation cycle of protein biosynthesis, the specific amino acid coded for by the mRNA is delivered by a complex that is comprised of the cognate aminoacyl-tRNA, elongation factor Tu and GTP. As this ternary complex binds to the ribosome, the anticodon end of the tRNA reaches the decoding center in the 30S subunit. Here we present the cryo- electron microscopy (EM) study of an Escherichia coli 70S ribosome-bound ternary complex stalled with an antibiotic, kirromycin. In the cryo-EM map the anticodon arm of the tRNA presents a new conformation that appears to facilitate the initial codon-anticodon interaction. Furthermore, the elbow region of the tRNA is seen to contact the GTPase associated center on the 50S subunit of the ribosome, suggesting an active role of the tRNA in the transmission of the signal prompting the GTP hydrolysis upon codon recognition. PMID- 12093759 TI - Health status and clinical practice: when will they meet? PMID- 12093760 TI - Clinical implications of cardiac ryanodine receptor/calcium release channel mutations linked to sudden cardiac death. PMID- 12093762 TI - Cardiology patient page. Atrial fibrillation. PMID- 12093761 TI - And the answer is, it doesn't much matter. PMID- 12093763 TI - Smooth muscle cells, but not myocytes, of host origin in transplanted human hearts. AB - BACKGROUND: There is increasing evidence to support a role for stem cells in the regeneration and repair of the human cardiovascular system. However, significant controversy still remains about the extent of chimerism present in blood vessels and myocytes of transplanted human hearts. METHODS AND RESULTS: We investigated the contribution of infiltrating host cells to human cardiac allografts by evaluating the origin of vascular smooth muscle cells and cardiac myocytes in hearts after orthotopic cardiac transplantation. Smooth muscle cells were identified in pathological human coronary artery specimens with antibodies against smooth muscle alpha-actin. DNA in situ hybridization for the human Y chromosome was then performed on the same samples to identify cells of male origin. Both myocytes and vascular smooth muscle cells were examined for the presence of the Y chromosome in sex-mismatched specimens. In positive control samples, 34.7% of nuclei contained a detectable Y chromosome; in sex-mismatched samples, 2.6% of the smooth muscle cells examined were of host origin. The Y chromosome in myocyte nuclei in male positive controls was detected; however, despite examination of >6000 myocyte nuclei in sex-mismatched specimens, we were unable to detect any nuclei with the clear presence of the Y chromosome. CONCLUSIONS: Vascular smooth muscle cells of infiltrating host cell origin can be found in human cardiac allografts. However, unlike prior reports, we found no evidence that chimerism is present in cardiac myocytes. PMID- 12093764 TI - Simvastatin reduces neointimal thickening in low-density lipoprotein receptor deficient mice after experimental angioplasty without changing plasma lipids. AB - BACKGROUND: Statins exert antiinflammatory and antiproliferative actions independent of cholesterol lowering. To determine whether these actions might affect neointimal formation, we investigated the effect of simvastatin on the response to experimental angioplasty in LDL receptor-deficient (LDLR-/-) mice, a model of hypercholesterolemia in which changes in plasma lipids are not observed in response to simvastatin. METHODS AND RESULTS: Carotid artery dilation (2.5 atm) and complete endothelial denudation were performed in male C57BL/6J LDLR-/- mice treated with low-dose (2 mg/kg) or high-dose (20 mg/kg) simvastatin or vehicle subcutaneously 72 hours before and then daily after injury. After 7 and 28 days, intimal and medial sizes were measured and the intima to media area ratio (I:M) was calculated. Total plasma cholesterol and triglyceride levels were similar in simvastatin- and vehicle-treated mice. Intimal thickening and I:M were reduced significantly by low- and high-dose simvastatin compared with vehicle alone. Simvastatin treatment was associated with reduced cellular proliferation (BrdU), leukocyte accumulation (CD45), and platelet-derived growth factor-induced phosphorylation of the survival factor Akt and increased apoptosis after injury. CONCLUSIONS: Simvastatin modulates vascular repair after injury in the absence of lipid-lowering effects. Although the mechanisms are not yet established, additional research may lead to new understanding of the actions of statins and novel therapeutic interventions for preventing restenosis. PMID- 12093765 TI - Interleukin-18 is a strong predictor of cardiovascular death in stable and unstable angina. AB - BACKGROUND: Interleukin (IL)-18 plays a central role in orchestrating the cytokine cascade and accelerates atherosclerosis and plaque vulnerability in animal models. However, epidemiological data evaluating the role of IL-18 levels in atherosclerosis are lacking. METHODS AND RESULTS: In a prospective study of 1229 patients with documented coronary artery disease, we measured baseline serum concentrations of IL-18 and other markers of inflammation. During the follow-up period (median, 3.9 years), 95 patients died of cardiovascular causes. Median serum concentrations of IL-18 were significantly higher among patients who had a fatal cardiovascular event than among those who did not (68.4 versus 58.7 pg/mL; P<0.0001). The hazard risk ratio of future cardiovascular death increased with increasing quartiles of IL-18 (hazard risk ratio, 1.46; 95% CI 1.21 to 1.76; P for trend <0.0001). After adjustment for most potential confounders, including the strong predictor ejection fraction as well as the inflammatory variables IL 6, high-sensitive C-reactive protein, and fibrinogen, this relation remained almost unchanged, such that patients within the highest quartile of IL-18 had a 3.3-fold increase in hazard risk compared with those in the first quartile (95% CI, 1.3 to 8.4, P=0.01). This relation was observed in patients with stable angina and patients with unstable angina at baseline. CONCLUSIONS: Serum IL-18 level was identified as a strong independent predictor of death from cardiovascular causes in patients with coronary artery disease regardless of the clinical status at admission. This result strongly supports recent experimental evidence of IL-18-mediated inflammation leading to acceleration and vulnerability of atherosclerotic plaques. PMID- 12093766 TI - Cardiomyocytes of noncardiac origin in myocardial biopsies of human transplanted hearts. AB - BACKGROUND: Cell replacement therapy with stem cells able to differentiate into cardiomyocytes has been discussed as a method for remodeling damaged myocardium. A physiological or pathophysiological situation in which this phenomenon might be relevant is not known. We studied the origin of cardiomyocytes in myocardial biopsies of male patients that had undergone sex-mismatched cardiac transplantation to determine whether cells containing a Y chromosome (and therefore being of recipient origin) are able to differentiate into cardiomyocytes. METHODS AND RESULTS: Myocardial biopsies (n=21) were obtained from the right ventricles of male patients (n=13) who had undergone sex mismatched heart transplantation. Tissue from 1 nontransplanted male and myocardial biopsies from sex-matched heart-transplanted patients served as controls. Cells from donor and recipient origins were identified by fluorescence in situ hybridization with the use of specific probes for X and Y chromosomes on paraffin sections of the biopsies. Cell types were identified by using immunostaining procedures on the same tissue sections. Cardiomyocytes of recipient origin were detected in 8 of 13 male recipients of female hearts. They were connected by gap junctions with adjacent myocytes. Of the cardiomyocyte nuclei, 0.16+/-0.04% (mean+/-SEM, median 0.09%) contained the Y-chromosomal marker. There was no detectable correlation with the extent or number of rejection episodes, time of transplantation, or medical treatment regimen. CONCLUSIONS: These results show that regeneration by cells of noncardiac origin (differentiated into cardiomyocytes and physiologically linked to neighboring myocytes) can be detected even in small myocardial biopsies. This may lead to new diagnostic and therapeutic strategies in the treatment of myocardial infarction, inflammatory heart disease, and/or heart failure. PMID- 12093767 TI - Stromal cell-derived factor-1alpha in unstable angina: potential antiinflammatory and matrix-stabilizing effects. AB - BACKGROUND: Chemokines play a pathogenic role in atherogenesis and plaque destabilization by activating and directing leukocytes into the atherosclerotic plaque. However, stromal cell-derived factor (SDF)-1 was recently found to have antiinflammatory effects, and we hypothesized that this chemokine could play a beneficial role in coronary artery disease. METHODS AND RESULTS: Plasma levels of SDF-1alpha were significantly decreased in patients with stable (n=30) and unstable angina (n=30) compared with healthy control subjects (n=20), particularly in those with unstable disease. By flow cytometry and RNase protection assay, we found decreased surface expression but increased gene expression of the SDF-1alpha receptor CXCR-4 in peripheral blood mononuclear cells (PBMC) from patients with stable angina and patients with unstable angina. In vitro, SDF-1alpha (500 ng/mL) reduced both unstimulated and endotoxin/mitogen stimulated mRNA and protein levels of monocyte chemoattractant protein-1, interleukin-8, matrix metalloproteinase-9, and tissue factor while increasing tissue inhibitor of metalloproteinases-1 in PBMC from patients with unstable angina. The SDF-1alpha-mediated suppression of monocyte chemoattractant protein-1 and interleukin-8 appears to involve cAMP/protein kinase A type I-dependent pathways. Finally, although SDF-1alpha suppressed the spontaneous release of these inflammatory mediators in unstable angina, enhancing effects were seen in unstimulated PBMC from healthy control subjects, possibly reflecting that PBMC in unstable angina are preactivated in vivo. CONCLUSIONS: In contrast to several other chemokines, our findings suggest that SDF-1alpha, at least in high concentrations, may mediate antiinflammatory and matrix-stabilizing effects in unstable angina. These effects may promote plaque stabilization, and therapeutic intervention that enhances SDF-1alpha activity could potentially be beneficial in acute coronary syndromes. PMID- 12093768 TI - Health status predicts long-term outcome in outpatients with coronary disease. AB - BACKGROUND: Although patient-reported health status measures have been used as end points in clinical trials, they are rarely used in other settings. Demonstrating that they independently predict mortality and hospitalizations among outpatients with coronary disease could emphasize their clinical value. METHODS AND RESULTS: This study evaluated the prognostic utility of the Seattle Angina Questionnaire (SAQ), a disease-specific health status measure for patients with coronary artery disease. Patients were enrolled in a prospective cohort study from 6 Veterans Affairs General Internal Medicine Clinics. All patients reporting coronary artery disease who completed a SAQ and had 1 year of follow-up were analyzed (n=5558). SAQ predictor variables were the physical limitation, angina stability, angina frequency, and quality-of-life scores. The primary outcome was 1-year all-cause mortality, and a secondary outcome was hospitalization for acute coronary syndrome (ACS). Lower SAQ scores were associated with increased risks of mortality and ACS admissions. Prognostic models controlling for demographic and clinical characteristics demonstrated significant independent mortality risk with lower SAQ physical limitation scores; odds ratios for mild, moderate, and severe limitation were 1.5, 2.0, and 4.0 versus minimal limitation (P<0.001). Odds ratios for mild, moderate, and severe angina frequency were 0.8, 1.2, and 1.6 (P=0.078). The odds ratios for ACS admission among those with mild, moderate, and severe angina frequency were 1.4, 2.0, and 2.2, respectively (P=0.016). CONCLUSIONS: SAQ scores are independently associated with 1-year mortality and ACS among outpatients with coronary disease and may serve a valuable role in the risk stratification of such patients. PMID- 12093769 TI - Quantitative assessment of intrinsic regional myocardial deformation by Doppler strain rate echocardiography in humans: validation against three-dimensional tagged magnetic resonance imaging. AB - BACKGROUND: Tissue Doppler echocardiography-derived strain rate and strain measurements (SDE) are new quantitative indices of intrinsic cardiac deformation. The aim of this study was to validate and compare these new indices of regional cardiac function to measurements of 3-dimensional myocardial strain by tagged MRI. METHODS AND RESULTS: The study population included 33 healthy volunteers, 17 patients with acute myocardial infarction, and 8 patients with suspected coronary artery disease who were studied during dobutamine stress echocardiography. Peak systolic myocardial velocities were measured by tissue Doppler echocardiography, peak systolic strain rates and strains by SDE, and strains by tagged MRI. In healthy individuals, longitudinal myocardial Doppler velocities decreased progressively from base to apex, whereas myocardial strain rates and strains were uniform in all segments. In patients with acute infarction, abnormal strains clearly identified dysfunctional areas. In infarcted regions, SDE showed 1.5+/ 4.3% longitudinal stretching compared with -15.0+/-3.9% shortening in remote myocardium (P<0.001), and radial measurements showed -6.9+/-4.1% thinning and 14.3+/-5.0% thickening (P<0.001), respectively. During dobutamine infusion, longitudinal strains by SDE increased significantly from -13.5% to -23.8% (P<0.01) and radial strains increased from 13.1+/-3.1% to 29.3+/-11.5% (P<0.01). Comparisons between myocardial strains by SDE and tagged MRI in healthy individuals (n=11), in infarct patients (n=17), and during stress echo (n=4) showed excellent correlations (r=0.89 and r=0.96 for longitudinal and radial strains, respectively, P< 0.001). CONCLUSIONS: The present study demonstrates the ability of Doppler echocardiography to measure myocardial strains in a clinical setting. Myocardial strains by Doppler may represent a new powerful method for quantifying left ventricular function noninvasively in humans. PMID- 12093770 TI - Rho-kinase mediates hypoxia-induced downregulation of endothelial nitric oxide synthase. AB - BACKGROUND: Hypoxia-induced pulmonary hypertension is a major cause of morbidity and mortality. Hypoxia induces pulmonary vasoconstriction, in part, by decreasing endothelial nitric oxide synthase (eNOS) expression. The mechanism by which hypoxia decreases eNOS expression is not known but may involve Rho-kinase-induced actin cytoskeletal changes in vascular endothelial cells. METHODS AND RESULTS: To determine whether hypoxia regulates eNOS expression through Rho-kinase, we exposed human saphenous and pulmonary artery endothelial cells to hypoxia (3% O2) with and without a Rho-kinase inhibitor, hydroxyfasudil (0.1 to 100 micromol/L), for various durations (0 to 48 hours). Hypoxia increased Rho-kinase expression and activity by 50% and 74%, decreased eNOS mRNA and protein expression by 66+/ 3% and 57+/-5%, and inhibited eNOS activity by 48+/-9%. All of these effects of hypoxia on eNOS were reversed by cotreatment with hydroxyfasudil. Furthermore, inhibition of Rho by Clostridium botulinum C3 transferase or Rho-kinase by overexpression of dominant-negative Rho-kinase reversed hypoxia-induced decrease in eNOS expression. Indeed, disruption of the actin cytoskeleton, the downstream target of Rho-kinase, by cytochalasin D also upregulated eNOS expression. Hypoxia reduced eNOS mRNA half-life from 22+/-2 to 13+/-2 hours, which was reversed by cotreatment with hydroxyfasudil. However, neither hypoxia nor hydroxyfasudil had any effects on eNOS gene transcription. CONCLUSIONS: These results indicate that hypoxia-induced decrease in eNOS expression is mediated by Rho-kinase and suggest that Rho-kinase inhibitors may have therapeutic benefits in patients with hypoxia induced pulmonary hypertension. PMID- 12093771 TI - Different metabolic predictors of white-coat and sustained hypertension over a 20 year follow-up period: a population-based study of elderly men. AB - BACKGROUND: The clinical significance of white-coat hypertension is still unclear. Moreover, no study has examined metabolic predictors of white-coat versus sustained hypertension. METHODS AND RESULTS: We investigated men (n=602) in a longitudinal population-based cohort who at age 70 years were identified as normotensive, white-coat hypertensive (office blood pressure [BP] > or =140/90 and daytime ambulatory BP <135/85 mm Hg), and sustained hypertensive (office BP > or =140/90 and daytime ambulatory BP > or =135/85 mm Hg). At baseline, when the subjects were aged 50 years, blood glucose, insulin, lipids, and fatty acid composition of the serum cholesterol esters were analyzed. The investigations at age 70 years included determination of insulin sensitivity and target organ damage. At age 50 years, individuals who 20 years later were identified as white coat hypertensive or sustained hypertensive showed significantly elevated BP, heart rate, and impaired glucose tolerance compared with normotensive subjects but white coat hypertensive subjects were leaner and had a more favorable serum cholesterol ester fatty acid profile than did sustained hypertensive subjects. At age 70 years, both white-coat and sustained hypertensive subjects showed an impaired insulin sensitivity, elevated blood glucose, and increased serum insulin and heart rate compared with normotensive subjects, but left ventricular mass and urinary albumin excretion were increased only in sustained hypertensive subjects. CONCLUSIONS: These findings indicate that although metabolic abnormalities and elevated heart rate were consistent over time in both hypertensive groups, a lower body mass index and more favorable dietary fat composition predicted the development of white-coat as opposed to sustained hypertension over 20 years. PMID- 12093772 TI - Clinical and molecular characterization of patients with catecholaminergic polymorphic ventricular tachycardia. AB - BACKGROUND: Mutations in the cardiac ryanodine receptor gene (RyR2) underlie catecholaminergic polymorphic ventricular tachycardia (CPVT), an inherited arrhythmogenic disease occurring in the structurally intact heart. The proportion of patients with CPVT carrying RyR2 mutations is unknown, and the clinical features of RyR2-CPVT as compared with nongenotyped CPVT are undefined. METHODS AND RESULTS: Patients with documented polymorphic ventricular arrhythmias occurring during physical or emotional stress with a normal heart entered the study. The clinical phenotype of the 30 probands and of 118 family members was evaluated, and mutation screening on the RyR2 gene was performed. Arrhythmias documented in probands were: 14 of 30 bidirectional ventricular tachycardia, 12 of 30 polymorphic ventricular tachycardia, and 4 of 30 catecholaminergic idiopathic ventricular fibrillation; RyR2 mutations were identified in 14 of 30 probands (36% bidirectional ventricular tachycardia, 58% polymorphic ventricular tachycardia, 50% catecholaminergic idiopathic ventricular fibrillation) and in 9 family members (4 silent gene carriers). Genotype-phenotype analysis showed that patients with RyR2 CPVT have events at a younger age than do patients with nongenotyped CPVT and that male sex is a risk factor for syncope in RyR2-CPVT (relative risk=4.2). CONCLUSIONS: CPVT is a clinically and genetically heterogeneous disease manifesting beyond pediatric age with a spectrum of polymorphic arrhythmias. beta-Blockers reduce arrhythmias, but in 30% of patients an implantable defibrillator may be required. Genetic analysis identifies two groups of patients: Patients with nongenotyped CPVT are predominantly women and become symptomatic later in life; patients with RyR2 CPVT become symptomatic earlier, and men are at higher risk of cardiac events. These data provide a rationale for prompt evaluation and treatment of young men with RyR2 mutations. PMID- 12093774 TI - Impaired carotid viscoelastic properties in women with polycystic ovaries. AB - BACKGROUND: The purpose of this study was to assess the elastic properties of the carotid arteries in women with polycystic ovarian syndrome, asymptomatic women with polycystic ovaries, and healthy controls. METHODS AND RESULTS: We recruited the following 60 subjects: 20 symptomatic women with polycystic ovaries attending the reproductive endocrinology clinics, 20 asymptomatic women with polycystic ovaries attending the family planning clinic, and 20 staff volunteers as healthy controls with normal ovaries on transvaginal scan. Compliance and stiffness index were assessed in the common and internal carotid arteries using duplex ultrasound equipped with an echo-locked arterial wall-tracking system. Compliance was significantly lower in the common carotid artery in symptomatic and asymptomatic women with polycystic ovaries than in the controls (10.7, 14.1, and 19.2 %mm Hg( 1)x10(-2), respectively). The arterial stiffness index was correspondingly increased (12.3, 10.2, and 6.7, respectively). Similar results were obtained in the internal carotid artery for compliance (10.1, 11.0, and 16.9 %mm Hg(-1)x10( 2), respectively) and stiffness index (14.8, 16.2, and 8.7, respectively). CONCLUSIONS: The results of this study provide additional evidence of vascular dysfunction in women with polycystic ovaries and are compatible with the hypothesis that they are at increased risk from coronary artery disease and stroke. PMID- 12093773 TI - Interventions on prevention of postoperative atrial fibrillation in patients undergoing heart surgery: a meta-analysis. AB - BACKGROUND: Postoperative atrial fibrillation (AF) is a common complication of cardiac surgery and has been associated with increased incidence of other complications and increased hospital length of stay (LOS). Prevention of AF is a reasonable clinical goal, and, consequently, many randomized trials have evaluated the effectiveness of pharmacological and nonpharmacological interventions for prevention of AF. To better understand the role of various prophylactic therapies against postoperative AF, a systematic review of evidence from randomized trials was performed. METHODS AND RESULTS: Fifty-two randomized trials (controlled by placebo or routine treatment) of beta-blockers, sotalol, amiodarone, or pacing were identified by systematic literature search. The 3 drug treatments each prevented AF with the following odds ratios (ORs): beta-blockers, 0.39 (95% CI, 0.28 to 0.52); sotalol, 0.35 (95% CI, 0.26 to 0.49); and amiodarone, 0.48 (95% CI, 0.37 to 0.61). Pacing was also effective; for biatrial pacing, the OR was 0.46 (95% CI, 0.30 to 0.71). The influence of pharmacological interventions on LOS was as follows: -0.66 day (95% CI, 2.04 to 0.72) for beta blockers; -0.40 day (95% CI, 0.87 to 0.08) for sotalol; and -0.91 day (95% CI, 1.59 to -0.23) for amiodarone. The influence for biatrial pacing was -1.54 day (95% CI, -2.85 to -0.24). The incidence of stroke was 1.2% in all the treatment groups combined and 1.4% in controls (OR, 0.90; 95% CI, 0.46 to 1.74). CONCLUSIONS: Beta-blockers, sotalol, and amiodarone all reduce risk of postoperative AF with no marked difference between them. There is evidence that use of these drugs will reduce LOS. Biatrial pacing is a promising new treatment opportunity. There was no evidence that reducing postoperative AF reduces stroke; however, data on stroke are incomplete. PMID- 12093775 TI - Stroke complicating percutaneous coronary interventions: incidence, predictors, and prognostic implications. AB - BACKGROUND: Stroke associated with percutaneous coronary intervention (PCI) is an infrequent although devastating complication. We investigated the incidence, predictors, and prognostic impact of periprocedural stroke in unselected patients undergoing PCI. METHODS AND RESULTS: A total of 9662 patients who underwent 12 407 PCIs between January 1990 and July 1999 were retrospectively studied. Stroke was diagnosed in 43 patients (0.38% of procedures). Patients with stroke were older (72+/-11 versus 64+/-11 years, P<0.001), had lower left ventricular ejection fraction (42+/-12 versus 46+/-13%, P=0.04) and more diabetes (39.5% versus 27.2%, P=0.07), and experienced a higher rate of intraprocedural complications necessitating emergency use of intra-aortic balloon pump (IABP) (23.3% versus 3.3%, P<0.001). In-hospital mortality (37.2% versus 1.1%, P<0.001) and 1-year mortality (56.1% versus 6.5%, P<0.001) were higher in patients with stroke. Compared with hemorrhagic stroke, patients with ischemic stroke had higher rate of in-hospital major adverse cardiac events (57.1% versus 25%, P=0.037). Multivariate logistic regression analysis identified emergency use of IABP as the strongest predictors for stroke (OR=9.6, CI 3.9 to 23.9, P<0.001), followed by prophylactic use of IABP (OR=5.1), age >80 years (OR=3.2, compared with age <50 years), and vein graft intervention (OR=2.7). CONCLUSIONS: Stroke associated with contemporary PCI is associated with substantial increased mortality. Elderly patients who experience intraprocedural complications necessitating the use of IABP are at particularly high risk. PMID- 12093776 TI - Neurohormonal activation and the chronic heart failure syndrome in adults with congenital heart disease. AB - BACKGROUND: Neurohormonal activation characterizes chronic heart failure, relates to outcome, and is a therapeutic target. It is not known whether a similar pattern of neurohormonal activation exists in adults with congenital heart disease and, if so, whether it relates to common measures of disease severity or whether cardiac anatomy is a better discriminant. METHODS AND RESULTS: Concentrations of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), endothelin-1 (ET-1), renin, aldosterone, norepinephrine, and epinephrine were determined in 53 adults with congenital heart disease, comprising 4 distinct anatomic subgroups (29 female; 33.5+/-1.5 years of age; New York Heart Association class 2.0+/-0.1, mean+/-SEM) and 15 healthy control subjects (8 female; 32.3+/-1.3 years of age). Systemic ventricular function was graded by a blinded echocardiographer as normal or mildly, moderately, or severely impaired. Adults with congenital heart disease had elevated levels of ANP (56.6 versus 3.1 pmol/L), BNP (35.8 versus 5.7 pmol/L), ET-1 (2.5 versus 0.7 pmol/L, all P<0.0001), renin (147 versus 16.3 pmol/L), norepinephrine (2.2 versus 1.6 pmol/L, both P<0.01) and aldosterone (546 versus 337 pmol/L, P<0.05). There was a highly significant stepwise increase in ANP, BNP, ET-1, and norepinephrine according to New York Heart Association class and systemic ventricular function, with even asymptomatic patients having evidence of significant neurohormonal activation. In contrast, there was no direct relationship between the 4 anatomic subgroups and any of the neurohormones studied. CONCLUSIONS: Neurohormonal activation in adult congenital heart disease bears the hallmarks of chronic heart failure, relating to symptom severity and ventricular dysfunction and not necessarily to anatomic substrate. Neurohormonal antagonism across this large and anatomically diverse population should be considered. PMID- 12093777 TI - Advanced coronary and carotid arteriopathy in young adults with childhood-onset chronic renal failure. AB - BACKGROUND: Cardiovascular mortality is excessive in young adults with end-stage renal disease (ESRD). The factors contributing to ESRD-related vascular disease are incompletely understood. Young adults with childhood-onset chronic renal failure (CRF) are uniquely suited for risk factor assessment because of their long-term exposure at an age when vascular pathology in the general population is still minimal. METHODS AND RESULTS: We used novel noninvasive technologies to screen for coronary and carotid artery disease in 39 patients with ESRD aged 19 to 39 years with childhood-onset CRF presently treated by dialysis or renal transplantation. Coronary artery calcification burden was assessed by CT scan with ECG gating and the intima-media thickness (IMT) of the carotid arteries by high-resolution ultrasound. Coronary artery calcifications were present in 92% of patients; calcium scores exceeded the 95th age- and sex-specific percentiles >10 fold on average. Carotid IMT was significantly increased compared with matched control subjects. Both coronary calcium scores and IMT were associated with cumulative dialysis and ESRD time and the cumulative serum calcium-phosphate product. Coronary calcium scores were strongly correlated with C-reactive protein and Chlamydia pneumoniae seropositivity, time-averaged mean serum parathyroid hormone, and plasma homocysteine. C-reactive protein and parathyroid hormone independently predicted coronary calcium accumulation. Smoking, obesity, and HbA1c were correlated with IMT in the control subjects but not in the patients. CONCLUSIONS: Young adults with childhood-onset CRF have a high prevalence of arteriopathy associated with indicators of microinflammation, hyperparathyroidism, calcium-phosphate overload, and hyperhomocysteinemia but not traditional atherogenic risk factors. These risk factors persist even after successful renal transplantation. PMID- 12093778 TI - Angiotensin II type 2 receptor overexpression preserves left ventricular function after myocardial infarction. AB - BACKGROUND: The role of the angiotensin II type 2 receptor (AT2-R) in left ventricular (LV) remodeling may depend on the underlying stimulus. We hypothesized that cardiac AT2-R overexpression in transgenic (TG) mice would attenuate remodeling after myocardial infarction (MI). METHODS AND RESULTS: Ten wild-type (WT) C57BL/6 mice and 12 TG mice that overexpress the AT2-R in the heart were studied by cardiac MRI at baseline and days 1, 7, and 28 post-MI induced by 1 hour of occlusion of the LAD followed by reperfusion. Short-axis imaging from apex to base was used to determine LV mass index, end-diastolic and end-systolic volume indices (EDVI, ESVI), regional wall thickness and thickening, and ejection fraction (EF). Gadolinium-DTPA was infused 20 minutes before day 1 imaging to assess infarct size. At baseline, heart rate, blood pressure, LV mass index, and EDVI were similar between groups. Baseline ESVI was lower (0.20+/-0.07 versus 0.45+/-0.15 microL/g, P<0.001) and EF higher (82.3+/-4.9% versus 67.7+/ 5.3%, P<0.001) in TG than WT. Infarct size was similar (36.6+/-7.2% in WT, 34.0+/ 7.8% in TG, P=NS). When controlled for baseline differences, ESVI was significantly less and EF significantly higher at all time points in TG versus WT. At day 28, ESVI was 1.05+/-0.32 microL/g in TG and 1.63+/-0.41 microL/g in WT, P<0.03, and EF was 47.3+/-5.8% versus 34.1+/-9.2%, P<0.003, respectively. Regional wall thickness and thickening were greater in TG both at baseline and at day 28. At day 28, blood pressure and LV dP/dt were higher in TG. CONCLUSIONS: Cardiac AT2-R overexpression improves LV systolic function at baseline and preserves function during post-MI remodeling. PMID- 12093779 TI - Mechanism of inducible regional dysfunction during dipyridamole stress. AB - BACKGROUND: We hypothesized that increased myocardial oxygen demand resulting from hypotension and reflex tachycardia unmasking a reduced endocardial myocardial blood flow (MBF) reserve is the mechanism of dipyridamole-induced regional dysfunction in chronic coronary artery disease. METHODS AND RESULTS: Ameroid constrictors were placed around the proximal coronary arteries and their major branches in 15 dogs to create chronic coronary stenosis. Seven days later, radiolabeled microsphere-derived MBF and 2-dimensional echocardiography-derived percent wall thickening (%WT) were measured at rest and after 0.56 mg/kg dipyridamole. Dipyridamole caused an increase (mean, 21%) in the rate-pressure product secondary to reflex tachycardia resulting from mild systemic hypotension. %WT in myocardial segments with an endocardial MBF reserve (dipyridamole/resting MBF) of 1.5 to 2.5 (n=35) did not change after dipyridamole, whereas it decreased in segments with an endocardial MBF reserve of <1.5 (n=30) and increased in those with an endocardial MBF reserve of > or =2.5 (n=45) (P<0.05). Most (80%) segments with endocardial MBF reserve of <1.5 and 14% with an endocardial MBF reserve of 1.5 to 2.5 showed inducible dysfunction after dipyridamole, whereas none of the segments with an endocardial MBF reserve of > or =2.5 showed this finding. A sigmoid relation (y=-6.74/[1+exp (19.9. [x-1.84])]+1.35. x, r=0.93, P<0.0001) was noted between endocardial MBF reserve and Delta%WT. In contrast, neither the epicardial MBF reserve nor the endocardial/epicardial MBF ratio during hyperemia was associated with inducible regional dysfunction. CONCLUSIONS: Increased myocardial oxygen demand resulting from hypotension and reflex tachycardia unmasking a reduced endocardial MBF reserve is the primary mechanism of dipyridamole-induced regional dysfunction in chronic coronary artery disease. PMID- 12093780 TI - Noninvasive measurement of myocardial activity concentrations and perfusion defect sizes in rats with a new small-animal positron emission tomograph. AB - BACKGROUND: We explored the feasibility of measuring regional tracer activity concentrations and flow defects in myocardium of rats with a high spatial resolution small-animal PET system (microPET). METHODS AND RESULTS: Myocardial images were obtained after intravenous (18)F-fluorodeoxyglucose (18FDG) in 11 normal rats (group 1) and assembled into polar maps. Regional 18F activity concentrations were measured in 9 regions of interest and compared with tissue activity concentrations measured by well counting. In another 9 rats (group 2), myocardial perfusion images were acquired with 13N-ammonia at baseline and during coronary occlusion. On the polar maps recorded during coronary occlusion, the size of perfusion defects was measured as the myocardium with <50% of maximum activity and expressed as percent total myocardium and was correlated with the area at risk defined by postmortem staining. The diagnostic quality of 18FDG and 13N-ammonia microPET images was good to excellent; the images were easily assembled into polar maps. In group 1, regional (18)F concentrations by microPET and postmortem were correlated linearly (r=0.99; P<0.01 for average and r=0.97; P<0.01 for regional concentrations). In group 2, perfusion defect sizes by microPET and postmortem were correlated linearly (P<0.01; r=0.93). CONCLUSIONS: The findings indicate the feasibility of noninvasive studies of the myocardium in rats with a dedicated small-animal PET-imaging device. PMID- 12093781 TI - Myocardial gene transfer and overexpression of beta2-adrenergic receptors potentiates the functional recovery of unloaded failing hearts. AB - BACKGROUND: Mechanical assistance of the failing left ventricle (LV) can lead to functional recovery after a period of unloading, including restoration of beta adrenergic receptor (betaAR) inotropic reserve. We tested whether prolonged LV unloading of failing rabbit hearts by use of a heterotopic transplantation technique could lead to recovery and whether adenoviral gene transfer of a beta2AR transgene (Adv-beta2AR) could alter this process. METHODS AND RESULTS: Heart failure was induced by coronary artery ligation in adult New Zealand White rabbits. After 4 weeks, failing hearts were heterotopically transplanted into recipient rabbits, allowing normal coronary perfusion but complete LV unloading. We also placed an LV latex balloon for remote access and in vivo physiological analysis. We found that there was reversal of signaling and functional abnormalities after 30 days of unloading. In another set of failing hearts, we randomly delivered, at the time of transplantation, either 2x10(11) viral particles of Adv-beta2AR or saline via the coronary arteries. Sham-operated animals with nonfailing hearts served as controls. After 5 days of unloading, in vivo LV contractility (LV dP/dt(max)) and relaxation (LV dP/dt(min)) were significantly decreased in saline-treated failing hearts compared with control nonfailing hearts (P<0.05). In failing hearts treated with Adv-beta2AR, however, LV dP/dt(max) and LV dP/dt(min) were improved in response to higher preloads (P<0.05) and betaAR stimulation (P<0.01). CONCLUSIONS: Heterotopic transplantation in the rabbit does allow recovery of the failing heart, and beta2AR overexpression acutely enhances this functional improvement. Accordingly, genetic manipulation of betaAR signaling may represent a novel molecular adjunct to mechanical assistance to facilitate functional myocardial recovery. PMID- 12093783 TI - Need to test the arterial inflammation hypothesis. PMID- 12093782 TI - Transforming growth factor-beta function blocking prevents myocardial fibrosis and diastolic dysfunction in pressure-overloaded rats. AB - BACKGROUND: Excessive myocardial fibrosis impairs cardiac function in hypertensive hearts. Roles of transforming growth factor (TGF)-beta in myocardial remodeling and cardiac dysfunction were examined in pressure-overloaded rats. METHODS AND RESULTS: Pressure overload was induced by a suprarenal aortic constriction in Wistar rats. Fibroblast activation (proliferation and phenotype transition to myofibroblasts) was observed after day 3 and peaked at days 3 to 7. Thereafter, myocyte hypertrophy and myocardial fibrosis developed by day 28. At day 28, echocardiography showed normal left ventricular fractional shortening, but the decreased ratio of early to late filling velocity of the transmitral Doppler velocity and hemodynamic measurement revealed left ventricular end diastolic pressure elevation, indicating normal systolic but abnormal diastolic function. Myocardial TGF-beta mRNA expression was induced after day 3, peaked at day 7, and remained modestly increased at day 28. An anti-TGF-beta neutralizing antibody, which was administered intraperitoneally daily from 1 day before operation, inhibited fibroblast activation and subsequently prevented collagen mRNA induction and myocardial fibrosis, but not myocyte hypertrophy. Neutralizing antibody reversed diastolic dysfunction without affecting blood pressure and systolic function. CONCLUSIONS: TGF-beta plays a causal role in myocardial fibrosis and diastolic dysfunction through fibroblast activation in pressure overloaded hearts. Our findings may provide an insight into a new therapeutic strategy to prevent myocardial fibrosis and diastolic dysfunction in pressure overloaded hearts. PMID- 12093784 TI - Images in cardiovascular medicine. Fetal diagnosis of right ventricular aneurysm associated with supraventricular tachycardia with left bundle-branch block aberrancy. PMID- 12093785 TI - Cardiovascular health in childhood: A statement for health professionals from the Committee on Atherosclerosis, Hypertension, and Obesity in the Young (AHOY) of the Council on Cardiovascular Disease in the Young, American Heart Association. PMID- 12093786 TI - Images in cardiovascular medicine. Cyclic tachycardia and hypotension. PMID- 12093788 TI - Interference with the cytoplasmic tail of gp210 disrupts "close apposition" of nuclear membranes and blocks nuclear pore dilation. AB - We tested the hypothesis that gp210, an integral membrane protein of nuclear pore complexes (NPCs), mediates nuclear pore formation. Gp210 has a large lumenal domain and small COOH-terminal tail exposed to the cytoplasm. We studied the exposed tail. We added recombinant tail polypeptides to Xenopus nuclear assembly extracts, or inhibited endogenous gp210 tails using anti-tail antibodies. Both strategies had no effect on the formation of fused flattened nuclear membranes, but blocked NPC assembly and nuclear growth. Inhibited nuclei accumulated gp210 and some nucleoporin p62, but failed to incorporate nup214/CAN, nup153, or nup98 and were defective for nuclear import of lamin B3. Scanning and transmission EM revealed a lack of "closely apposed" inner and outer membranes, and the accumulation of novel arrested structures including "mini-pores." We conclude that gp210 has early roles in nuclear pore formation, and that pore dilation is mediated by gp210 and its tail-binding partner(s). We propose that membrane fusion and pore dilation are coupled, acting as a mechanism to control nuclear pore size. PMID- 12093791 TI - Homocysteine is a protein amino acid in humans. Implications for homocysteine linked disease. AB - Homocysteine is thought to be a non-protein amino acid. However, in vitro studies suggest that homocysteine is likely to be incorporated by indirect mechanisms into proteins in living organisms. Here I show that homocysteine is a protein amino acid in humans. Homocysteine bound by amide or peptide linkages (Hcy-N protein) is present in human hemoglobin, serum albumin, and gamma-globulins. 1 molecule of homocysteine per 1000 or 1670 molecules of methionine was present in hemoglobin or albumin, respectively. Other proteins, such as low density lipoprotein, high density lipoprotein, transferrin, antitrypsin, and fibrinogen, contained lower amounts of Hcy-N-protein. In human plasma, levels of Hcy-N protein represented from 0.3 to 23% of total homocysteine. Thus, Hcy-N-protein is a significant component of homocysteine metabolism in humans, possibly contributing to adverse effects of homocysteine on human cells. PMID- 12093789 TI - A novel mechanism for the regulation of amyloid precursor protein metabolism. AB - Modifier of cell adhesion protein (MOCA; previously called presenilin [PS] binding protein) is a DOCK180-related molecule, which interacts with PS1 and PS2, is localized to brain areas involved in Alzheimer's disease (AD) pathology, and is lost from the soluble fraction of sporadic Alzheimer's disease (AD) brains. Because PS1 has been associated with gamma-secretase activity, MOCA may be involved in the regulation of beta-amyloid precursor protein (APP) processing. Here we show that the expression of MOCA decreases both APP and amyloid beta peptide secretion and lowers the rate of cell-substratum adhesion. In contrast, MOCA does not lower the secretion of amyloid precursor-like protein (APLP) or several additional type 1 membrane proteins. The phenotypic changes caused by MOCA are due to an acceleration in the rate of intracellular APP degradation. The effect of MOCA expression on the secretion of APP and cellular adhesion is reversed by proteasome inhibitors, suggesting that MOCA directs nascent APP to proteasomes for destruction. It is concluded that MOCA plays a major role in APP metabolism and that the effect of MOCA on APP secretion and cell adhesion is a downstream consequence of MOCA-directed APP catabolism. This is a new mechanism by which the expression of APP is regulated. PMID- 12093792 TI - A novel PAAD-containing protein that modulates NF-kappa B induction by cytokines tumor necrosis factor-alpha and interleukin-1beta. AB - PAAD domains are found in diverse proteins of unknown function and are structurally related to a superfamily of protein interaction modules that includes death domains, death effector domains, and Caspase activation and recruitment domains. Using bioinformatics strategies, cDNAs were identified that encode a novel protein of 110 kDa containing a PAAD domain followed by a putative nucleotide-binding (NACHT) domain and several leucine-rich repeat domains. This protein thus resembles Cryopyrin, a protein implicated in hereditary hyperinflammation syndromes, and was termed PAN2 for PAAD and NACHT-containing protein 2. When expressed in HEK293 cells, PAN2 suppressed NF-kappaB induction by the cytokines tumor necrosis factor-alpha (TNFalpha) and interleukin-1beta (IL 1beta), suggesting that this protein operates at a point of convergence in these two cytokine signaling pathways. This PAN2-mediated suppression of NF-kappaB was evident both in reporter gene assays that measured NF-kappaB transcriptional activity and electromobility shift assays that measured NF-kappaB DNA binding activity. PAN2 also suppressed NF-kappaB induction resulting from overexpression of several adapter proteins and protein kinases involved in the TNF or IL-1 receptor signal transduction, including TRAF2, TRAF6, RIP, IRAK2, and NF-kappaB inducing kinase as well as the IkappaB kinases IKKalpha and IKKbeta. PAN2 also inhibited the cytokine-mediated activation of IKKalpha and IKKbeta as measured by in vitro kinase assays. Furthermore, PAN2 association with IKKalpha was demonstrated by co-immunoprecipitation assays, suggesting a direct effect on the IKK complex. These observations suggest a role for PAN2 in modulating NF-kappaB activity in cells, thus providing the insights into the potential functions of PAAD family proteins and their roles in controlling inflammatory responses. PMID- 12093790 TI - Dok-related protein negatively regulates T cell development via its RasGTPase activating protein and Nck docking sites. AB - Downstream of kinase (Dok)-related protein (DokR, also known as p56(dok)/FRIP/Dok R) is implicated in cytokine and immunoreceptor signaling in myeloid and T cells. Tyrosine phosphorylation induces DokR to bind the signal relay molecules, RasGTPase-activating protein (RasGAP) and Nck. Here, we have examined the function of DokR during hematopoietic development and the requirement for RasGAP and Nck binding sites in its biological function. Retroviral-mediated expression of DokR in bone marrow cells dramatically inhibited their capacity to form colonies in vitro in response to the cytokines macrophage colony-stimulating factor and stem cell factor, whereas responses to interleukin-3 and granulocyte macrophage colony-stimulating factor were only weakly affected. When introduced into lethally irradiated mice, hematopoietic cells expressing DokR showed a drastically reduced capacity to repopulate lymphoid tissues. Most notably, DokR dramatically reduced repopulation of the thymus, in part by reducing the number of T cell precursors seeding in the thymus, but equally, through inhibiting the transition of CD4(-)CD8(-) to CD4(+)CD8(+) T cells. Consequently, the number of mature peripheral T cells was markedly reduced. In contrast, a minimal effect on B cell and myeloid lineage development was observed. Importantly, functional RasGAP and Nck binding sites were found to be essential for the biological effects of DokR in vitro and in vivo. PMID- 12093793 TI - Biosynthesis of tunicamycin and metabolic origin of the 11-carbon dialdose sugar, tunicamine. AB - Tunicamycin is a reversible inhibitor of polyprenol-phosphate: N-acetylhexosamine 1-phosphate translocases and is produced by several Streptomyces species. We have examined tunicamycin biosynthesis, an important but poorly characterized biosynthetic pathway. Biosynthetic precursors have been identified by incorporating radioactive and stable isotopes, and by determining the labeling pattern using electrospray ionization-collision induced dissociation-mass spectrometry (ESI-CID-MS), and proton, deuterium, and C-13 nuclear magnetic resonance (NMR) spectroscopy. Preparation and analysis of [uracil-5-(2)H]-labeled tunicamycin established the complete ESI-CID-MS fragmentation pathway for the major components of the tunicamycin complex. Competitive metabolic experiments indicate that 7 deuteriums incorporate into tunicamycin from [6,6'-(2)H,(2)H] labeled D-glucose, 6 of which arise from D-GlcNAc and 1 from uridine and/or D ribose. Inverse correlation NMR experiments (heteronuclear single-quantum coherence (HSQC)) of (13)C-labeled tunicamycin enriched from D-[1-(13)C]glucose suggest that the unique tunicamine 11-carbon dialdose sugar backbone arises from a 5-carbon furanose precursor derived from uridine and a 6-carbon N-acetylamino pyranose precursor derived from UDP-D-N-acetylglucosamine. The equivalent incorporation of (13)C into both the alpha-1" and beta-11' anomeric carbons of tunicamycin supports a direct biosynthesis via 6-carbon metabolism. It also indicates that the tunicamine motif and the alpha-1"-linked GlcNAc residue are both derived from the same metabolic pool of UDP-GlcNAc, without significant differential metabolic processing. A biosynthetic pathway is therefore proposed for tunicamycin for the first time: an initial formation of the 11-carbon tunicamine sugar motif from uridine and UDP-GlcNAc via uridine-5'-aldehyde and UDP-4-keto-6-ene-N-acetylhexosamine, respectively, and subsequent formation of the anomeric-to-anomeric alpha, beta-1",11'-glycosidic bond. PMID- 12093794 TI - Critical role of NADPH oxidase-derived reactive oxygen species in generating Ca2+ oscillations in human aortic endothelial cells stimulated by histamine. AB - There is increasing evidence that intracellular reactive oxygen species (ROS) play a role in cell signaling and that the NADPH oxidase is a major source of ROS in endothelial cells. At low concentrations, agonist stimulation of membrane receptors generates intracellular ROS and repetitive oscillations of intracellular Ca(2+) concentration ([Ca(2+)](i)) in human endothelial cells. The present study was performed to examine whether ROS are important in the generation or maintenance of [Ca(2+)](i) oscillations in human aortic endothelial cells (HAEC) stimulated by histamine. Histamine (1 microm) increased the fluorescence of 2',7'-dihydrodichlorofluorescin diacetate in HAEC, an indicator of ROS production. This was partially inhibited by the NADPH oxidase inhibitor diphenyleneiodonium (DPI, 10 microm), by the farnesyltransferase inhibitor H Ampamb-Phe-Met-OH (2 microm), and in HAEC transiently expressing Rac1(N17), a dominant negative allele of the protein Rac1, which is essential for NADPH oxidase activity. In indo 1-loaded HAEC, 1 microm histamine triggered [Ca(2+)](i) oscillations that were blocked by DPI or H-Ampamb-Phe-Met-OH. Histamine stimulated [Ca(2+)](i) oscillations were not observed in HAEC lacking functional Rac1 protein but were observed when transfected cells were simultaneously exposed to a low concentration of hydrogen peroxide (10 microm), which by itself did not alter either [Ca(2+)](i) or levels of inositol 1,4,5-trisphosphate (Ins-1,4,5 P(3)). Thus, histamine generates ROS in HAEC at least partially via NADPH oxidase activation. NADPH oxidase-derived ROS are critical to the generation of [Ca(2+)](i) oscillations in HAEC during histamine stimulation, perhaps by increasing the sensitivity of the endoplasmic reticulum to Ins-1,4,5-P(3). PMID- 12093795 TI - The catalytic center of glucose-6-phosphatase. HIS176 is the nucleophile forming the phosphohistidine-enzyme intermediate during catalysis. AB - Glucose-6-phosphatase (G6Pase), a key enzyme in glucose homeostasis, is anchored to the endoplasmic reticulum by nine transmembrane helices. The amino acids comprising the catalytic center of G6Pase include Lys(76), Arg(83), His(119), Arg(170), and His(176). During catalysis, a His residue in G6Pase becomes phosphorylated generating an enzyme-phosphate intermediate. It was predicted that His(176) would be the amino acid that acts as a nucleophile forming a phosphohistidine-enzyme intermediate, and His(119) would be the amino acid that provides the proton needed to liberate the glucose moiety. However, the phosphate acceptor in G6Pase has eluded molecular characterization. To identify the His residue that covalently bound the phosphate moiety, we generated recombinant adenoviruses carrying G6Pase wild type and active site mutants. A 40-kDa [(32)P]phosphate-G6Pase intermediate was identified after incubating [(32)P]glucose 6-phosphate with microsomes expressing wild type but not with microsomes expressing either H119A or H176A mutant G6Pase. Human G6Pase contains five methionine residues at positions 1, 5, 121, 130, and 279. After cyanogen bromide cleavage, His(119) is predicted to be within a 116-amino acid peptide of 13.5 kDa with an isoelectric point of 5.3 (residues 6-121), and His(176) is predicted to be within a 149-amino acid peptide of 16.8 kDa with an isoelectric point of 9.3 (residues 131-279). We show that after digestion of a non glycosylated [(32)P]phosphate-G6Pase intermediate by cyanogen bromide, the [(32)P]phosphate remains bound to a peptide of 17 kDa with an isoelectric point above 9, demonstrating that His(176) is the phosphate acceptor in G6Pase. PMID- 12093796 TI - The serine protease plasmin triggers expression of MCP-1 and CD40 in human primary monocytes via activation of p38 MAPK and janus kinase (JAK)/STAT signaling pathways. AB - The mechanism of proinflammatory activation of human monocytes by plasmin is unknown. Here we demonstrate that in human primary monocytes, plasmin stimulates mitogen-activated protein kinase (MAPK) signaling via phosphorylation of MAPK kinase 3/6 (MKK3/6) and p38 MAPK that triggers subsequent DNA binding of transcription factor activator protein-1 (AP-1). The AP-1 complex contained phosphorylated c-Jun and ATF2, and its DNA binding activity was blocked by the p38 MAPK inhibitor SB203580. In addition, plasmin elicits Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling, as detected by phosphorylation of JAK1 tyrosine kinase and STAT1 and STAT3 proteins. Plasmin-induced DNA binding of STAT1 and STAT3 was blocked by SB203580 and AG490, inhibitors of p38 MAPK and JAK, respectively, but not by U0126, an inhibitor of MKK1/2. DNA binding of NF-kappaB remained unaffected by any of these inhibitors. The plasmin-induced signaling led to expression of monocyte chemoattractant protein-1 (MCP-1) and CD40, which required activation of both p38 MAPK and JAK/STAT signaling pathways. Additionally, signaling through both p38 MAPK and JAK is involved in the plasmin-mediated monocyte migration, whereas the formylmethionylleucylphenylalanine-induced chemotaxis remained unaffected. Taken together, our data demonstrate a novel function of the serine protease plasmin in a proinflammatory signaling network. PMID- 12093797 TI - Inhibition of cytokine-induced matrix metalloproteinase 9 expression by peroxisome proliferator-activated receptor alpha agonists is indirect and due to a NO-mediated reduction of mRNA stability. AB - Rat renal mesangial cells express high levels of matrix metalloproteinase 9 (MMP 9) in response to inflammatory cytokines such as interleukin 1beta (IL-1beta). We tested whether ligands of the peroxisome proliferator-activated receptor (PPARalpha) could influence the cytokine-induced expression of MMP-9. Different PPARalpha agonists dose-dependently inhibited the IL-1beta-triggered increase in gelatinolytic activity mainly by decreasing the MMP-9 steady-state mRNA levels. PPARalpha agonists on their own had no effects on MMP-9 mRNA levels and gelatinolytic activity. Surprisingly, the reduction of MMP-9 mRNA levels by PPARalpha activators contrasted with an amplification of cytokine-mediated MMP-9 gene promoter activity and mRNA expression. The potentiation of MMP-9 promoter activity functionally depends on an upstream peroxisome proliferator-responsive element-like binding site, which displayed an increased DNA binding of a PPARalpha immunopositive complex. In contrast, the IL-1beta-induced DNA-binding of nuclear factor kappaB was significantly impaired by PPARalpha agonists. Most interestingly, in the presence of an inducible nitric-oxide synthase (iNOS) inhibitor, the PPARalpha-mediated suppression switched to a strong amplification of IL-1beta-triggered MMP-9 mRNA expression. Concomitantly, activators of PPARalpha potentiated the cytokine-induced iNOS expression. Using actinomycin D, we found that NO, but not PPARalpha activators, strongly reduced the stability of MMP-9 mRNA. In contrast, the stability of MMP-9 protein was not affected by PPARalpha activators. In summary, our data suggest that the inhibitory effects of PPARalpha agonists on cytokine-induced MMP-9 expression are indirect and primarily due to a superinduction of iNOS with high levels of NO reducing the half-life of MMP-9 mRNA. PMID- 12093798 TI - Autoinhibition and isoform-specific dominant negative inhibition of the type II cGMP-dependent protein kinase. AB - In the absence of cyclic nucleotides, the cAMP-dependent protein kinase and cGMP dependent protein kinases (cGKs) suppress phosphotransfer activity at the catalytic cleft by competitive inhibition of substrate binding with a pseudosubstrate sequence within the holoenzyme. The magnitude of inhibition can be diminished by autophosphorylation near this pseudosubstrate sequence. Activation of type I cGK (cGKI) and type II cGK (cGKII) are differentially regulated by their cyclic nucleotide-binding sites. To address the possibility that the distinct activation mechanisms of cGKII and cGKI result from differences in the autophosphorylation of the inhibitory domain, we investigated the effects of autophosphorylation on the kinetics of activation. Unlike the type I cGKs (cGKIalpha and Ibeta), cGKII autophosphorylation did not alter the basal activity, nor the sensitivity of the enzyme to cyclic nucleotide activation. To determine residues responsible for autoinhibition of cGKII, Ala was substituted for basic residues (Lys(122), Arg(118), and Arg(119)) or a hydrophobic residue (Val(125)) within the putative pseudosubstrate domain of cGKII. The integrity of these residues was essential for full cGKII autoinhibition. Furthermore, a cGKII truncation mutant containing this autoinhibitory region demonstrated a nanomolar IC(50) toward a constitutively active form of cGKII. Finally, we present evidence that the dominant negative properties of this truncation mutant are specific to cGKII when compared with cAMP-dependent protein kinase Calpha and cGKIbeta. These findings extend the known differences in the activation mechanisms among cGK isoforms and allow the design of an isoform-specific cGKII inhibitor. PMID- 12093799 TI - Normal thyroid thermogenesis but reduced viability and adiposity in mice lacking the mitochondrial glycerol phosphate dehydrogenase. AB - The mitochondrial glycerol phosphate dehydrogenase (mGPD) is important for metabolism of glycerol phosphate for gluconeogenesis or energy production and has been implicated in thermogenesis induced by cold and thyroid hormone treatment. mGPD in combination with the cytosolic glycerol phosphate dehydrogenase (cGPD) is proposed to form the glycerol phosphate shuttle, catalyzing the interconversion of dihydroxyacetone phosphate and glycerol phosphate with net oxidation of cytosolic NADH. We made a targeted deletion in Gdm1 and produced mice lacking mGPD. On a C57BL/6J background these mice showed a 50% reduction in viability compared with wild-type littermates. Uncoupling protein-1 mRNA levels in brown adipose tissue did not differ between mGPD knockout and control pups, suggesting normal thermogenesis. Pups lacking mGPD had decreased liver ATP and slightly increased liver glycerol phosphate. In contrast, liver and muscle metabolites were normal in adult animals. Adult mGPD knockout animals had a normal cold tolerance, normal circadian rhythm in body temperature, and demonstrated a normal temperature increase in response to thyroid hormone. However, they were found to have a lower body mass index, a 40% reduction in the weight of white adipose tissue, and a slightly lower fasting blood glucose than controls. The phenotype may be secondary to consequences of the obligatory production of cytosolic NADH from glycerol metabolism in the mGPD knockout animal. We conclude that, although mGPD is not essential for thyroid thermogenesis, variations in its function affect viability and adiposity in mice. PMID- 12093800 TI - Lethal hypoglycemic ketosis and glyceroluria in mice lacking both the mitochondrial and the cytosolic glycerol phosphate dehydrogenases. AB - The activities of either the mitochondrial or cytosolic glycerol phosphate dehydrogenase (mGPD, cGPD) plus that of glycerol kinase are required for the use of glycerol in aerobic metabolism and gluconeogenesis. A knockout mouse lacking mGPD has reduced body weight and fertility but shows remarkably normal liver and muscle metabolite levels. The BALB/cHeA mouse strain, which lacks cGPD, breeds well and is phenotypically normal, although it demonstrates metabolite abnormalities in certain tissues. Crosses were made between these two strains, and mice were generated that lacked both dehydrogenases. These mice, although active and nursing well for several days, failed to grow, and usually died within the first week. Liver glycerol phosphate levels were elevated 30-fold, whereas liver ATP, ADP, and AMP levels were reduced by 30-40%. Plasma glycerol was elevated 30- to 50-fold to 30-50 mm, and urine glycerol exceeded 0.45 m (4% w/v). GPD-deficient mice were hypoglycemic, had a 50% increase in plasma free fatty acids, and developed ketonuria within the first day of life. Uncoupling protein-1 mRNA in brown adipose tissue was reduced 60%. These mice share some features of both glycerol kinase deficiency and hereditary fructose intolerance, suggesting the phenotype may be due to the combined effects of the loss of a gluconeogenic substrate, the osmotic effects of glycerol, and the metabolic effects of the accumulation of a phosphorylated metabolite. PMID- 12093801 TI - Glycogen synthase kinase 3beta is activated by cAMP and plays an active role in the regulation of melanogenesis. AB - In human and mouse, cAMP plays a key role in the control of pigmentation. cAMP, through the activation of protein kinase A, increases the expression of microphthalmia-associated transcription factor (MITF), which in turn stimulates tyrosinase gene expression, to allow melanin synthesis. Beyond this simplified scheme, cAMP inhibits phosphatidylinositol 3-kinase (PI3K), and inhibition of PI3K, by a specific inhibitor, stimulates melanogenesis. However, the link between the PI3K pathway and melanogenesis remained to be elucidated. In this report, we showed that cAMP, through a protein kinase A-independent mechanism, led to inhibition of AKT phosphorylation and activity. Consistent with the role of AKT in the regulation of glycogen synthase kinase 3beta (GSK3beta), cAMP decreased the phosphorylation of GSK3beta and stimulated its activity. Further, experiments were performed to investigate the role of GSK3beta in the regulation of MITF expression and function. We observed that GSK3beta regulated neither MITF promoter activity nor the intrinsic transcriptional activity of MITF but synergized with MITF to activate the tyrosinase promoter. Additionally, lithium, a GSK3beta inhibitor, impaired the response of the tyrosinase promoter to cAMP, and cAMP increased the binding of MITF to the M-box. Taking into account that GSK3beta phosphorylates MITF and increases the ability of MITF to bind its target sequence, our results indicate that activation of GSK3beta by cAMP facilitates MITF binding to the tyrosinase promoter, thereby leading to stimulation of melanogenesis. PMID- 12093802 TI - Mitochondrial alterations induced by the p13II protein of human T-cell leukemia virus type 1. Critical role of arginine residues. AB - Human T-cell leukemia virus type 1 encodes a number of "accessory" proteins of unclear function; one of these proteins, p13(II), is targeted to mitochondria and disrupts mitochondrial morphology. The present study was undertaken to unravel the function of p13(II) through (i) determination of its submitochondrial localization and sequences required to alter mitochondrial morphology and (ii) an assessment of the biophysical and biological properties of synthetic peptides spanning residues 9-41 (p13(9-41)), which include the amphipathic mitochondrial targeting sequence of the protein. p13(9-41) folded into an alpha helix in micellar environments. Fractionation and immunogold labeling indicated that full length p13(II) accumulates in the inner mitochondrial membrane. p13(9-41) induced energy-dependent swelling of isolated mitochondria by increasing inner membrane permeability to small cations (Na(+), K(+)) and released Ca(2+) from Ca(2+) preloaded mitochondria. These effects as well as the ability of full-length p13(II) to alter mitochondrial morphology in cells required the presence of four arginines, forming the charged face of the targeting signal. The mitochondrial effects of p13(9-41) were insensitive to cyclosporin A, suggesting that full length p13(II) might alter mitochondrial permeability through a permeability transition pore-independent mechanism, thus distinguishing it from the mitochondrial proteins Vpr and X of human immunodeficiency virus type 1 and hepatitis B virus, respectively. PMID- 12093803 TI - Redox regulation of plant homeodomain transcription factors. AB - Several families of plant transcription factors contain a conserved DNA binding motif known as the homeodomain. In two of these families, named Hd-Zip and glabra2, the homeodomain is associated with a leucine zipper-like dimerization motif. A group of Hd-Zip proteins, namely Hd-ZipII, contain a set of conserved cysteines within the dimerization motif and adjacent to it. Incubation of one of these proteins, Hahb-10, in the presence of thiol-reducing agents such as dithiothreitol or reduced glutathione produced a significant increase in DNA binding. Under such conditions, the protein migrated as a monomer in non-reducing SDS-polyacrylamide gels. Under oxidizing conditions, a significant proportion of the protein migrated as dimers, suggesting the formation of intermolecular disulfide bonds. A similar behavior was observed for the glabra2 protein HAHR1, which also contains two conserved cysteines within its dimerization domain. Site directed mutagenesis of the cysteines to serines indicated that each of them has different roles in the activation of the proteins. Purified thioredoxin was able to direct the NADPH-dependent activation of Hahb-10 and HAHR1 in the presence of thioredoxin reductase. The results suggest that redox conditions may operate to regulate the activity of these groups of plant transcription factors within plant cells. PMID- 12093804 TI - Suppression of estrogen receptor-mediated transcription and cell growth by interaction with TR2 orphan receptor. AB - The transcriptional activity of the estrogen receptor (ER) is known to be highly modulated by the character and amount of coregulator proteins present in the cells. TR2 orphan receptor (TR2), a member of the nuclear receptor superfamily without identified ligands, is found to be expressed in the breast cancer cell lines and to function as a repressor to suppress ER-mediated transcriptional activity. Utilizing an interaction blocker, ER-6 (amino acids 312-340), responsible for TR2 interaction, the suppression of ER by TR2 could be reversed, suggesting that this suppression is conferred by the direct protein-protein interaction. Administration of antisense TR2, resulting in an enhancement of ER transcriptional activity in MCF7 cells, indicates that endogenous TR2 normally suppresses ER-mediated signaling. To gain insights into the molecular mechanism by which TR2 suppresses ER, we found that TR2 could interrupt ER DNA binding via formation of an ER-TR2 heterodimer that disrupted the ER homodimerization. The suppression of ER transcription by TR2 consequently caused the inhibition of estrogen-induced cell growth and G(1)/S transition in estrogen-dependent breast cancer cells. Taken together in addition to the potential roles in spermatogenesis and neurogenesis, our data provide a novel biological function of TR2 that may exert an important repressor in regulating ER activity in mammary glands. PMID- 12093805 TI - Role of AP-1 in the coordinate induction of rat glutamate-cysteine ligase and glutathione synthetase by tert-butylhydroquinone. AB - GSH synthesis occurs via two enzymatic steps catalyzed by glutamate-cysteine ligase (GCL, made up of two subunits) and GSH synthetase (GS). Recently, we described coordinate induction of GCL subunits and GS. To study GS transcriptional regulation, we have cloned and characterized a 2.2-kb 5'-flanking region of the rat GS (GenBank accession number AF333982). One transcriptional start site is located at 51 nucleotides upstream of the translational start site. The rat GS promoter drove efficiently luciferase expression in H4IIE cells. Sequential deletion analysis revealed DNA regions that are involved in positive and negative regulation. One repressor identified was NF1. tert-Butylhydroquinone (TBH) exerted a dose- and time-dependent increase in the mRNA level and promoter activity of both GCL subunits and GS. TBH increased protein binding to several regions of the GS promoter, c-jun expression, and activator protein 1 (AP-1) binding activity to several of the putative AP-1-binding sites of the GS promoter. Blocking AP-1 binding with dominant-negative c-jun led to decreased basal expression and significantly blocked the TBH-induced increase in promoter activity and mRNA level of all three genes. In conclusion, AP-1 is required for basal expression of GCL and GS; while NF1 serves as a repressor of GS, increased AP-1 transactivation is the predominant mechanism for coordinate induction of GCL and GS expression by TBH. PMID- 12093806 TI - A selective interaction between OS-9 and the carboxyl-terminal tail of meprin beta. AB - OS-9, a protein previously uncharacterized, was shown to interact specifically with the intracellular region of the membrane proteinase meprin beta found in brush border membranes of kidney and small intestine. We have shown previously that this cytoplasmic region is indispensable for the maturation of meprin beta, which included an endoplasmic reticulum (ER)-to-Golgi translocation. We characterized OS-9 and found that it is associated with ER membranes and that it is exposed to the cytoplasm. Consistent with the kinetics of maturation of meprin beta, OS-9 associates with meprin beta only transiently, coinciding with ER-to Golgi transport of meprin beta. The OS-9-binding site in the cytoplasmic domain of meprin beta overlaps the region essential for this transport. We characterized alternatively spliced forms of rat and mouse OS-9, and we found that only the non spliced form of OS-9 binds to meprin beta, implicating the spliced out segment in the binding, and suggesting the possible mechanism of the regulation of OS-9 function. Taken together, our results indicated that OS-9 may be involved in the ER-to-Golgi transport of meprin beta. Ubiquitous expression of OS-9 raises the possibility that it may interact with other membrane proteins that possess the cytoplasmic moiety homologous to that of meprin beta during their ER-to-Golgi transition. PMID- 12093807 TI - Cardiac troponin T isoforms affect the Ca2+ sensitivity and inhibition of force development. Insights into the role of troponin T isoforms in the heart. AB - At least four isoforms of troponin T (TnT) exist in the human heart, and they are expressed in a developmentally regulated manner. To determine whether the different N-terminal isoforms are functionally distinct with respect to structure, Ca(2+) sensitivity, and inhibition of force development, the four known human cardiac troponin T isoforms, TnT1 (all exons present), TnT2 (missing exon 4), TnT3 (missing exon 5), and TnT4 (missing exons 4 and 5), were expressed, purified, and utilized in skinned fiber studies and in reconstituted actomyosin ATPase assays. TnT3, the adult isoform, had a slightly higher alpha-helical content than the other three isoforms. The variable region in the N terminus of cardiac TnT was found to contribute to the determination of the Ca(2+) sensitivity of force development in a charge-dependent manner; the greater the charge the higher the Ca(2+) sensitivity, and this was primarily because of exon 5. These studies also demonstrated that removal of either exon 4 or exon 5 from TnT increased the cooperativity of the pCa force relationship. Troponin complexes reconstituted with the four TnT isoforms all yielded the same maximal actin tropomyosin-activated myosin ATPase activity. However, troponin complexes containing either TnT1 or TnT2 (both containing exon 5) had a reduced ability to inhibit this ATPase activity when compared with wild type troponin (which contains TnT3). Interestingly, fibers containing these isoforms also showed less relaxation suggesting that exon 5 of cardiac TnT affects the ability of Tn to inhibit force development and ATPase activity. These results suggest that the different N-terminal TnT isoforms would produce different functional properties in the heart that would directly affect myocardial contraction. PMID- 12093808 TI - Nucleotide binding states of hsp70 and hsp90 during sequential steps in the process of glucocorticoid receptor.hsp90 heterocomplex assembly. AB - A minimal system of five purified proteins, hsp90, hsp70, Hop, hsp40, and p23, assembles glucocorticoid receptor (GR).hsp90 heterocomplexes and causes the simultaneous opening of the steroid binding cleft to access by steroid. The first step in assembly is the ATP-dependent and hsp40 (YDJ-1)-dependent binding of hsp70 to the GR, which primes the receptor for subsequent ATP-dependent activation by hsp90, Hop, and p23 (Morishima, Y., Murphy, P. J. M., Li, D. P., Sanchez, E. R., and Pratt, W. B. (2000) J. Biol. Chem. 275, 18054-18060). Here we have examined the nucleotide-bound states of the two essential chaperones in each step. We show that it is the ATP-bound state of hsp70 that interacts initially with the GR. After rapid priming and washing, the primed GR.hsp70 complex rapidly binds hsp90 in the second step reaction in a nucleotide-independent manner. The rate-limiting step is the ATP-dependent opening of the steroid binding cleft after hsp90 binding. This activating step requires the N-terminal ATP-binding site of hsp90, but we cannot establish any role for a C-terminal ATP-binding site in steroid binding cleft opening. The reported specific inhibitors of the C terminal ATP site on hsp90 inhibit the generation of steroid binding, but they have other effects in this multiprotein system that could explain the inhibition. PMID- 12093809 TI - Convergence of the target of rapamycin and the Snf1 protein kinase pathways in the regulation of the subcellular localization of Msn2, a transcriptional activator of STRE (Stress Response Element)-regulated genes. AB - The subcellular localization of Msn2, a transcriptional activator of STRE (stress response element)-regulated genes, is modulated by carbon source availability. In cells growing in glucose, Msn2 is located mainly in the cytosol, whereas in carbon source-starved cells, Msn2 is located largely inside the nucleus. However, in cells lacking Reg1 (the regulatory subunit of the Reg1/Glc7 protein phosphatase complex), the regulation of subcellular distribution is absent, Msn2 being constitutively present in the cytosol. The localization defect in these mutants is specific for carbon starvation stress, and it is because of the presence of an abnormally active Snf1 protein kinase that inhibits the nuclear localization of Msn2 upon carbon starvation. Active Snf1 kinase is also able to avoid the effects of rapamycin, a drug that by inhibiting the TOR kinase pathway leads to a nuclear localization of Msn2 in wild type cells. Therefore, active Snf1 and the TOR kinase pathway may affect similar cytosolic steps in the regulation of the subcellular localization of Msn2. PMID- 12093810 TI - Gliadin T cell epitope selection by tissue transglutaminase in celiac disease. Role of enzyme specificity and pH influence on the transamidation versus deamidation process. AB - Tissue transglutaminase (TG2) can modify proteins by transamidation or deamidation of specific glutamine residues. TG2 has a major role in the pathogenesis of celiac disease as it is both the target of disease-specific autoantibodies and generates deamidated gliadin peptides that are recognized by CD4(+), DQ2-restricted T cells from the celiac lesions. Capillary electrophoresis with fluorescence-labeled gliadin peptides was used to separate and quantify deamidated and transamidated products. In a competition assay, the affinity of TG2 to a set of overlapping gamma-gliadin peptides was measured and compared with their recognition by celiac lesion T cells. Peptides differed considerably in their competition efficiency. Those peptides recognized by intestinal T cell lines showed marked competition indicating them as excellent substrates for TG2. The enzyme fine specificity of TG2 was characterized by synthetic peptide libraries and mass spectrometry. Residues in positions -1, +1, +2, and +3 relative to the targeted glutamine residue influenced the enzyme activity, and proline in position +2 had a particularly positive effect. The characterized sequence specificity of TG2 explained the variation between peptides as TG2 substrates indicating that the enzyme is involved in the selection of gluten T cell epitopes. The enzyme is mainly localized extracellularly in the small intestine where primary amines as substrates for the competing transamidation reaction are present. The deamidation could possibly take place in this compartment as an excess of primary amines did not completely inhibit deamidation of gluten peptides at pH 7.3. However, lowering of the pH decreased the reaction rate of the TG2-catalyzed transamidation, whereas the rate of the deamidation reaction was considerably increased. This suggests that the deamidation of gluten peptides by TG2 more likely takes place in slightly acidic environments. PMID- 12093811 TI - Identification of cytoplasmic domains within the epithelial Na+ channel reactive at the plasma membrane. AB - The activity of membrane proteins is controlled, in part, by protein-protein interactions localized to the plasma membrane. In the current study, domains within the epithelial Na(+) channel (ENaC) reactive at the plasma membrane were identified using a novel yeast one-hybrid screen. The cytosolic N terminus of alphaENaC and the cytosolic C termini of alpha-, beta-, and gammaENaC contained domains reactive at the plasma membrane. Fluorescent micrographs of epithelial cells overexpressing fusion proteins of enhanced green fluorescent protein and mENaC cytosolic domains were consistent with those in yeast. A novel membrane reactive domain within the cytosolic C terminus of gamma-mENaC was localized to the 17 amino acids between residues Thr(584)-Pro(600). Two overlapping internalization signals within the C terminus of gamma-mENaC, a WW-binding domain (PY motif) and a tyrosine-based endocytic signal, were additive with respect to decreasing complementation and expression levels of hybrid proteins. Decreases in expression levels of hybrid proteins containing the PY and endocytic motif were reversed with latrunculin A, an inhibitor of endosomal movement. Decreases in complementation and expression levels of hybrid proteins mediated by the combined PY and overlapping endocytic motif proceeded in the absence of established ubiquitination sites within ENaC. In addition, the endocytic motif was active in the absence of the PY motif, demonstrating that these two domains, while possibly interacting, also have discrete functions. The novel domains within the cytosolic N terminus of alphaENaC and the C termini of alpha-, beta-, and gammaENaC identified here are likely to be involved in protein-protein and/or protein-lipid interactions localized to the plasma membrane. We hypothesize that these newly identified domains play a role in modulating ENaC activity. PMID- 12093813 TI - Antigenic and functional properties of the human red blood cell urea transporter hUT-B1. AB - The Kidd (JK) blood group locus encodes the urea transporter hUT-B1, which is expressed on human red blood cells and other tissues. The common JK*A/JK*B blood group polymorphism is caused by a single nucleotide transition G838A changing Asp 280 to Asn-280 on the polypeptide, and transfection of erythroleukemic K562 cells with hUT-B1 cDNAs carrying either the G838 or the A838 nucleotide substitutions resulted in the isolation of stable clones that expressed the Jk(a) or Jk(b) antigens, respectively, thus providing the first direct demonstration that the hUT-B1 gene encodes the Kidd blood group antigens. In addition, immunochemical analysis of red blood cells demonstrated that hUT-B1 also exhibits ABO determinants attached to the single N-linked sugar chain at Asn-211. Moreover, immunoadsorption studies, using inside-out and right-side-out red cell membrane vesicles as competing antigen, demonstrated that the C- and N-terminal ends of hUT-B1 are oriented intracellularly. Mutagenesis and functional studies by expression in Xenopus oocytes revealed that both cysteines Cys-25 and Cys-30 (but not alone) are essential for plasma membrane addressing. Conversely, the transport function was not affected by the JK*A/JK*B polymorphism, C-terminal deletion (residues 360-389), or mutation of the extracellular N-glycosylation consensus site and remains poorly para-chloromercuribenzene sulfonate (pCMBS) sensitive. However, transport studies by stopped flow light scattering using Jk K562 transfectants demonstrated that the hUT-B1-mediated urea transport is pCMBS sensitive in an erythroid context, as reported previously for the transporter of human red blood cells. Mutagenesis analysis also indicated that Cys-151 and Cys 236, at least alone, are not involved in pCMBS inhibition. Altogether, these antigenic, topologic, and functional properties might have implications into the physiology of hUT-B1 and other members of the urea transporter family. PMID- 12093812 TI - Molecular determinants of permeation through the cation channel TRPV4. AB - We have studied the molecular determinants of ion permeation through the TRPV4 channel (VRL-2, TRP12, VR-OAC, and OTRPC4). TRPV4 is characterized by both inward and outward rectification, voltage-dependent block by Ruthenium Red, a moderate selectivity for divalent versus monovalent cations, and an Eisenman IV permeability sequence. We identify two aspartate residues, Asp(672) and Asp(682), as important determinants of the Ca(2+) sensitivity of the TRPV4 pore. Neutralization of either aspartate to alanine caused a moderate reduction of the relative permeability for divalent cations and of the degree of outward rectification. Neutralizing both aspartates simultaneously caused a much stronger reduction of Ca(2+) permeability and channel rectification and additionally altered the permeability order for monovalent cations toward Eisenman sequence II or I. Moreover, neutralizing Asp(682) but not Asp(672) strongly reduces the affinity of the channel for Ruthenium Red. Mutations to Met(680), which is located at the center of a putative selectivity filter, strongly reduced whole cell current amplitude and impaired Ca(2+) permeation. In contrast, neutralizing the only positively charged residue in the putative pore region, Lys(675), had no obvious effects on the properties of the TRPV4 channel pore. Our findings delineate the pore region of TRPV4 and give a first insight into the possible architecture of its permeation pathway. PMID- 12093814 TI - Oxidation of methoxyphenethylamines by cytochrome P450 2D6. Analysis of rate limiting steps. AB - Cytochrome P450 (P450) 2D6 is involved in the oxidation of a large fraction ( approximately 30%) of drugs used by humans and also catalyzes the O-demethylation of the model substrates 3- and 4-methoxyphenethylamine followed by subsequent ring hydroxylation to dopamine. Burst kinetics were not observed; rate-limiting step(s) must occur prior to product formation. Rates of reduction of ferric P450 2D6 were stimulated by 3- or 4-methoxyphenethylamine or the inhibitor quinidine; reduction is not the most rate-limiting step. The non-competitive intramolecular deuterium isotope effect, an estimate of the intrinsic isotope effect, for 4 methoxyphenethylamine O-demethylation was 9.6. Intermolecular non-competitive deuterium isotope effects of 3.1-3.8 were measured for k(cat) and k(cat)/K(m) for both O-demethylation reactions, implicating at least partially rate-limiting C-H bond breaking. Simulation of steady-state kinetic data yielded a catalytic mechanism dominated by the rates of (i) Fe(2+)O(2)(-) protonation (plus O-O bond scission) and (ii) C-H bond breaking, consistent with the appearance of the spectral intermediates in the steady state, attributed to iron-oxygen complexes. However, all the rates of individual steps (or rates of combined steps) are considerably higher than k(cat), and the contributions of several steps must be considered in understanding rates of the P450 2D6 reactions. PMID- 12093815 TI - Noncatalytic domains of RGS9-1.Gbeta 5L play a decisive role in establishing its substrate specificity. AB - The complex between the photoreceptor-specific regulator of G protein signaling (RGS) protein, RGS9-1, and type 5 G protein beta-subunit, Gbeta5L, regulates the duration of the cellular response to light by stimulating the GTPase activity of G protein, transducin. An important property of RGS9-1.Gbeta5L is that it interacts specifically with transducin bound to its effector, cGMP phosphodiesterase, rather than with transducin alone. The minimal structure within the RGS9-1.Gbeta5L complex capable of activating transducin GTPase is the catalytic domain of RGS9. This domain itself is also able to discriminate between free and effector-bound transducin but to a lesser degree than RGS9-1.Gbeta5L. The goal of this study was to determine whether other, noncatalytic domains of RGS9-1.Gbeta5L enhance the intrinsic specificity of the catalytic domain or whether they set the specificity of RGS9-1.Gbeta5L regardless of the specificity of its catalytic domain. We found that a double L353E/R360P amino acid substitution reversed the specificity of the recombinant catalytic domain but did not reverse the specificity of RGS9-1.Gbeta5L. However, the degree of discrimination between free and effector-bound transducin was reduced. Therefore, noncatalytic domains of RGS9-1.Gbeta5L play a decisive role in establishing its substrate specificity, yet the high degree of this specificity observed under physiological conditions requires an additional contribution from the catalytic domain. PMID- 12093816 TI - Evidence for a lectin activity for human interleukin 3 and modeling of its carbohydrate recognition domain. AB - We demonstrate that human interleukin 3 (IL-3) is a lectin recognizing specifically the glycosaminoglycan part of a chondroitin sulfate proteoglycan (PGS3; Normand, G., Kuchler, S., Meyer, A., Vincendon, G., and Zanetta, J. P. (1988) J. Neurochem. 51, 665-676) isolated from the adult rat brain. The specificity of the interaction of this particular proteoglycan with IL-3 is due to the abundance of GlcA(2S)beta 1,3GalNAc(4S)beta 1 disaccharide units as suggested by (1)H NMR. Computational docking experiments of the lower energy conformers of the different disaccharides from chondroitin sulfates reveal a privileged binding site for GlcA(2S)beta 1,3GalNAc(4S)beta 1 (involving His-26, Arg-29, Asn-70, and Trp-104) localized in an area of IL-3 different from the receptor-binding domain previously identified by others (Bagley, C. J., Phillips, J., Cambareri, B., Vadas, M. A., and Lopez, A. F. (1996) J. Biol. Chem. 271, 31922-31928). Molecular modeling of the mutation P33G, described as increasing the biological activity of IL-3 without affecting its receptor binding (Lokker, N. A., Movva, N. R., Strittmatter, U., Fagg, B., and Zenke, G. (1991) J. Biol. Chem. 266, 10624-10631) provokes a change of the three-dimensional structure of IL-3, especially in the area of the putative carbohydrate recognition domain defined above. Computational docking experiments of the different disaccharides of chondroitin sulfates indicate a loss of affinity for the previous ligand but a higher affinity for the classic disaccharide of chondroitin-4-sulfate. This change from a rare and specific ligand to a more abundant constituent of proteoglycans could induce an increased quantitative association between the IL-3 receptors and its ligands and, consequently, an increased signaling. PMID- 12093817 TI - Transfer of M2 muscarinic acetylcholine receptors to clathrin-derived early endosomes following clathrin-independent endocytosis. AB - Upon agonist stimulation, many G protein-coupled receptors such as beta(2) adrenergic receptors are internalized via beta-arrestin- and clathrin-dependent mechanisms, whereas others, like M(2) muscarinic acetylcholine receptors (mAChRs), are internalized by clathrin- and arrestin-independent mechanisms. To gain further insight into the mechanisms that regulate M(2) mAChR endocytosis, we investigated the post-endocytic trafficking of M(2) mAChRs in HeLa cells and the role of the ADP-ribosylation factor 6 (Arf6) GTPase in regulating M(2) mAChR internalization. Here, we report that M(2) mAChRs are rapidly internalized by a clathrin-independent pathway that is inhibited up to 50% by expression of either GTPase-defective Arf6 Q67L or an upstream Arf6 activator, Galpha(q) Q209L. In contrast, M(2) mAChR internalization was not affected by expression of dominant negative dynamin 2 K44A, which is a known inhibitor of clathrin-dependent endocytosis. Nevertheless, M(2) mAChRs, which are initially internalized in structures that lack clathrin-dependent endosomal markers, quickly localize to endosomes that contain the clathrin-dependent, early endosomal markers early endosome autoantigen-1, transferrin receptor, and GTPase-defective Rab5 Q79L, which is known to swell early endosomal compartments. These results suggest that M(2) mAChRs initially internalize via an Arf6-associated, clathrin-independent pathway but then quickly merge with the clathrin endocytic pathway at the level of early endosomes. PMID- 12093818 TI - Transcription factor Sp1 phosphorylation induced by shear stress inhibits membrane type 1-matrix metalloproteinase expression in endothelium. AB - Membrane type 1-matrix metalloproteinase (MT1-MMP) plays a key role in endothelial cell migration, matrix remodeling, and angiogenesis. Previous studies demonstrated that a mechanical force, cyclic strain, increases MT1-MMP expression by displacing Sp1 with increased Egr-1 expression and binding to the promoter site. However, the effect of shear stress (SS) on MT1-MMP expression is poorly understood. Although Egr-1 mRNA transcription and protein was induced (7.6-fold) in response to SS (n = 5, 0-8 h, p < 0.05), SS decreased MT1-MMP mRNA transcription and protein levels in a time-dependent fashion (10, 50, and 90% reduction at 1, 4, and 8 h, respectively; n = 5, p < 0.05). Egr-1 protein was increased after SS and cyclic strain, but Sp1 was serine-phosphorylated only after SS. SS increased Sp1 DNA binding (3.8-, 5.8-, and 2.4-fold increase at 1, 4, and 8 h, respectively; n = 5, p < 0.05) that was inhibitable by calf intestinal phosphatase. Thus, SS inhibits MT1-MMP expression despite Egr-1 up regulation by inducing the serine phosphorylation of Sp1, which in turn increases its binding affinity for its site on the MT1-MMP promoter, reducing the ability of Egr-1 to displace it. These data illustrate the complex control of microvascular endothelial cell MT1-MMP expression in response to distinct environmental stimuli (cyclic strain versus shear stress), consisting of both the modulation of specific transcription factor expression (Egr-1) as well as transcription factor post-translational modification (serine phosphorylation of Sp1). PMID- 12093820 TI - The use of urinary gonadotrophins should be discouraged. AB - In view of concerns regarding the potential presence and infectivity of prion proteins in human urinary gonadotrophin preparations, together with the availability of both recombinant FSH and recombinant LH, it is argued that the use of urinary gonadotrophins should be discouraged. PMID- 12093819 TI - The C2 domain of phosphatidylserine decarboxylase 2 is not required for catalysis but is essential for in vivo function. AB - Phosphatidylserine decarboxylase 2 (Psd2p) is currently being used to study lipid trafficking processes in intact and permeabilized yeast cells. The Psd2p contains a C2 homology domain and a putative Golgi retention/localization (GR) domain. C2 domains play important functions in membrane binding and docking reactions involving phospholipids and proteins. We constructed a C2 domain deletion variant (C2Delta) and a GR deletion variant (GRDelta) of Psd2p and examined their effects on in vivo function and catalysis. Immunoblotting confirmed that the predicted immature and mature forms of Psd2(C2Delta)p, Psd2(GRDelta)p, and wild type Psd2p were produced in vivo and that the proteins localized normally. Enzymology revealed that the Psd2(C2Delta)p and Psd2(GRDelta)p were catalytically active and could readily be expressed at levels 10-fold higher than endogenous Psd2p. Both Psd2p and Psd2(GRDelta)p expression complemented the growth defect of psd1Deltapsd2Delta strains and resulted in normal aminoglycerophospholipid metabolism. In contrast, the Psd2(C2Delta)p failed to complement psd1Deltapsd2Delta strains, and [(3)H]serine labeling revealed a severe defect in the formation of PtdEtn in both intact and permeabilized cells, indicative of disruption of lipid trafficking. These findings identify an essential, non catalytic function of the C2 domain of Psd2p and raise the possibility that it plays a direct role in membrane docking and/or PtdSer transport to the enzyme. PMID- 12093821 TI - Is there a risk of prion disease after the administration of urinary-derived gonadotrophins? AB - Concern has been raised recently about the possibility of prion proteins appearing in the urine of animals and, possibly, humans affected by prion disease [scrapie, bovine spongiform encephalopathy (BSE) and Creutzfeldt Jakob disease (CJD)]. A debate has started in which the suggestion has been made that the purification of human urine for the provision of gonadotrophins should be discontinued. The alternative would be to use recombinantly-derived gonadotrophin preparations. The recombinant products, however, rely upon bovine serum during the cell culture process and could potentially also be exposed to abnormal prion proteins. It is reassuring that the different types of gonadotrophin preparations that are currently available are produced with either urine or bovine serum that is sourced from countries that at the present time appear to be free of BSE and new variant CJD. We can therefore be reassured that the gonadotrophins that we use therapeutically appear to be equally safe. PMID- 12093822 TI - Risk of infection is not the main problem. AB - The risk of infection from prion proteins in urinary preparations of human gonadotrophins is uncertain--and is of lesser importance than the risk of multiple pregnancies and issues of cost. PMID- 12093823 TI - The conflict between effective and affordable health care--a perspective from the developing world. AB - The recent introduction of recombinant FSH into the clinical management of patients suffering from infertility appears to be associated with several treatment benefits when compared with urinary human menopausal gonadotrophin. However, from the perspective of the developing world the associated increase in cost is a cause for concern--particularly if the "cheaper" urinary gonadotrophins are no longer marketed. The need for infertility care in Africa is significant, but health resources are very limited. The commonest cause of infertility in Africa is tubal disease, so that assisted reproductive techniques, and therefore exogenous gonadotrophins, are central to effective management. The conflict between affordable and effective health care is addressed. PMID- 12093824 TI - Investigation of the infertile couple: a one-stop outpatient endoscopy-based approach. AB - Transvaginal hydrolaparoscopy (THL) is a new culdoscopic technique for exploration of the pelvic cavity that takes advantage of micro-endoscopic technology and uses aquaflotation for inspection of the tubo-ovarian structures. In infertility patients, THL is systematically combined with mini-hysteroscopy, chromopertubation, fimbrioscopy and, when indicated, salpingoscopy. Mini hysteroscopy in combination with the chromopertubation test allows accurate assessment of the uterine cavity and tubal patency. The transvaginal access combined with the aquaflotation during THL facilitates detailed inspection of the tubo-ovarian structures and detection of subtle pelvic disease. This combined transvaginal endoscopic approach allows complete evaluation of the reproductive tract. THL is better tolerated than hysterosalpingography, less invasive than standard laparoscopy, and can be used safely as a first line investigation of the female partner in a one-stop infertility clinic. PMID- 12093825 TI - Tubal ectopic pregnancy: a patho-physiological explanation involving endometriosis. AB - The condition of tubal ectopic pregnancy is presented from diverse points of view, bringing out physiological explanations for its occurrence in primates and striking absence in other mammals. Part of the flexibility underlying ectopic pregnancy in humans stems from the absence of a uterine luteolytic mechanism, enabling early embryonic development in the Fallopian tube without compromising function of the corpus luteum. Attention is devoted to a potential overlap between the composition of tubal and uterine fluids, and to specific mixing between the two fluid compartments, expressed in an ability of the human oocyte or zygote to tolerate transplantation to the uterus. Perturbed tubal oocyte transport is seen as a contributory factor, not least as a sequel to episodes of infection and a modified endosalpinx, but the essay then reasons strongly for an involvement of endometriosis in the aetiology of tubal ectopic pregnancy. Proliferation of refluxed endometrial tissue arrested within the Fallopian tube could provide the epithelial characteristics of a uterine environment. Accordingly, an experimental model is proposed for tubal ectopic pregnancy in animals based upon transplants of endometrial tissue and the subsequent introduction of embryos into both the Fallopian tubes and uterus; the latter would suppress the luteolytic mechanism. Finally, advances are suggested based upon molecular scanning of human ectopic tissues and those derived from animal models. If molecular probes could be developed to detect either early tubal pregnancy or a propensity to this pathology, such advances would clearly have clinical relevance-not least in view of an enhanced incidence of tubal pregnancy arising after assisted reproduction technology. PMID- 12093826 TI - Infant feeding with soy formula milk: effects on the testis and on blood testosterone levels in marmoset monkeys during the period of neonatal testicular activity. AB - BACKGROUND: This study has addressed concerns about possible effects of feeding human infants soy formula milk (SFM). METHODS: This is a feeding study in marmosets, using a mainly co-twin design. From 4-5 until 35-45 days of age, co twin males were fed by hand with either standard (cow) formula milk (SMA = controls) or with SFM for approximately 8 h each day (2 h at weekends) and intake related to bodyweight. Blood samples were collected at 18-20 and at 35-45 days of age in 13 sets of co-twins plus two non-twin males per group and, at the later age, seven sets of co-twins were killed and the testes and pituitary gland fixed for cell counts. RESULTS: Weight gain and formula intake were similar in both feeding groups. SMA-fed males had mean testosterone levels of 2.8-3.1 ng/ml, typical of the "neonatal testosterone rise", whereas SFM-fed males exhibited consistently lower mean levels (1.2-2.6 ng/ml); paired comparison in SMA-and SFM fed co-twins at day 35-45 revealed 53-70% lower levels in 11 of 13 co-twins fed with SFM (P = 0.004). Further evidence for suppression of testosterone levels in SFM-fed males came from comparison of the frequency of low testosterone levels (<0.5 ng/ml). In historical controls aged 35-45 days, two out of 22 values were <0.5 ng/ml, a similar frequency as found in control SMA-fed males (one out of 15 values <0.5 ng/ml). In contrast, 12 out of 15 values for SFM-fed males were <0.5 ng/ml (P < 0.001). There was no consistent relationship between SFM intake/g and testosterone levels. Paradoxically, the mean number of Leydig cells per testis was increased by 74% (P < 0.001) in co-twins fed SFM, when compared with their SMA-fed brothers, whereas no significant changes were found in numbers of Sertoli and germ cells. Because of the lack of gonadotrophin assays, the number of immunopositive LHbeta and FSHbeta cells in the pituitary gland, and their ratio, were determined but no consistent difference was found between SMA- and SFM-fed twins. CONCLUSIONS: Based on the average isoflavone content of the SFM brand used, intake of isoflavones was estimated at 1.6-3.5 mg/kg/day in the SFM-fed marmosets which is 40-87% of that reported in 4 month human infants fed on a 100% SFM diet. It is therefore considered likely that similar, or larger, effects to those shown here in marmosets may occur in human male infants fed with SFM. Whether the changes described result in longer-term effects is under investigation. PMID- 12093827 TI - Endometriosis is associated with an altered profile of intestinal microflora in female rhesus monkeys. AB - BACKGROUND: The intestinal microflora provide a strong defence against intestinal pathogens, and may be altered in inflammatory conditions that impact the gut, such as endometriosis. Therefore, intestinal bacteria shed from rhesus monkeys with endometriosis were compared with age-matched healthy controls. A second study assessed the prevalence of intestinal inflammation in female monkeys to determine whether endometriosis is associated with an increased likelihood of intestinal inflammation. METHODS: Differential and selective agars were used to enumerate total and Gram-negative aerobic and facultatively anaerobic bacteria, as well as Lactobacilli, from female monkeys with or without endometriosis. In addition, the prevalence of intestinal inflammation in monkeys with or without endometriosis was determined in a retrospective analysis of necropsy reports. RESULTS: Monkeys with endometriosis had a significantly different profile of shed microflora. Endometriosis was associated with lower Lactobacilli concentrations and higher Gram-negative bacteria concentrations. Moreover, there was a higher prevalence of intestinal inflammation in monkeys with endometriosis in comparison to healthy controls. CONCLUSIONS: Endometriosis is associated with an altered profile of intestinal microflora in rhesus monkeys. Although the exact mechanisms linking endometriosis and the microflora are unknown, it is possible that the microflora were affected by endometriosis-associated intestinal inflammation. PMID- 12093828 TI - Different mechanisms for the induction of copper-zinc superoxide dismutase and manganese superoxide dismutase by progesterone in human endometrial stromal cells. AB - BACKGROUND: The present study was undertaken to investigate the cAMP-dependent regulation of copper-zinc superoxide dismutase (Cu,Zn-SOD) and manganese SOD (Mn SOD) by ovarian steroids in human endometrial stromal cells (ESC). METHODS AND RESULTS: To examine the effect of cAMP on SOD expression, ESC were incubated with dibutyryl-cAMP (db-cAMP, 0.5 mmol/l), forskolin (25 micromol/l), or estradiol (E(2), 10(-8) mol/l) + medroxyprogesterone acetate (MPA, 10(-6) mol/l), for 18 days. E(2) + MPA significantly increased Cu,Zn-SOD activity and mRNA concentrations, whereas db-cAMP and forskolin had no effect. On the other hand, Mn-SOD activity and mRNA concentration were significantly increased by all of these treatments. Insulin-like growth factor-binding protein-1, a marker of decidualization, was clearly induced by db-cAMP, forskolin or E(2) + MPA, accompanied by morphological changes characteristic of decidualization. To study whether the increase in Mn-SOD by db-cAMP or E(2) + MPA was mediated by cAMP dependent protein kinase A (PKA), ESC were incubated with protein kinase inhibitor (PKI) (10 microg/ml), an inhibitor of PKA, in the presence of db-cAMP or E(2) + MPA. The increase in Mn-SOD activity following db-cAMP or E(2) + MPA was completely inhibited by PKI. CONCLUSIONS: In the process of decidualization, E(2) + MPA increases Mn-SOD expression via a cAMP-dependent pathway. Cu,Zn-SOD is also up-regulated by E(2) + MPA, but via a different pathway from that involving cAMP. PMID- 12093829 TI - Investigation of suppression of the hypothalamic-pituitary-gonadal axis to restore spermatogenesis in azoospermic men treated for childhood cancer. AB - BACKGROUND: Does suppression of the hypothalamic-pituitary-gonadal (HPG) axis restore spermatogenesis in men rendered azoospermic following treatment of childhood cancer? METHODS: Seven men with azoospermia secondary to treatment for childhood cancer, median age (range), 22.2 (18-25.3) years, aged 10.4 (4.4-13.3) years at original diagnosis, participated. Each subject underwent semen analysis and testicular biopsy, followed by treatment with medroxyprogesterone acetate (MPA), 300 mg i.m. repeated after 12 weeks, with 800 mg testosterone pellets s.c. on day 1 to suppress the HPG axis. Hormone and semen analysis was performed every 6 weeks for 48 weeks. A second testicular biopsy was performed at week 48. RESULTS: Before HPG axis suppression, mean +/- SEM plasma LH was 9.0 +/- 1.8 U/l, testosterone 17.9 +/- 1.5 nmol/l and FSH 22.4 +/- 4.4 U/l. Median (range) venous plasma and seminal plasma inhibin B levels were 10.0 (7.8-35) and 11.2 (7.8-770) ng/l respectively. During HPG suppression, FSH and LH levels were undetectable for > or =12 weeks followed by a gradual return to pretreatment concentrations by 48 weeks. All men remained azoospermic at study completion and complete absence of germ cells on biopsies was demonstrated by immunocytochemistry for all specimens pre- and post-HPG axis suppression. CONCLUSIONS: HPG axis suppression with MPA-testosterone for > or =12 weeks did not restore spermatogenesis in azoospermic men treated with gonadotoxic radiotherapy and chemotherapy for childhood cancer. PMID- 12093830 TI - The concentration of inhibin B in follicular fluid: relation to oocyte maturation and embryo development. AB - BACKGROUND: The objective of the present study was to investigate the correlation between inhibin B and estradiol levels in follicular fluid (FF) with the quality of subsequent embryo development from in-vitro fertilized oocytes aspirated from the same follicle. METHODS: A total of 156 infertile women undergoing controlled ovarian stimulation for IVF and embryo transfer was recruited to the present study. Prospectively, 233 FF samples and matched mature oocytes were studied. Concentrations of inhibin B and estradiol were determined by enzyme-linked immunosorbent assay (ELISA) and immunofluorometric assay (IFMA) respectively. RESULTS: Inhibin B levels in FF were significantly correlated with embryo scores on days 2 and 3 (48 and 72 h after oocyte retrieval). In contrast, both inhibin B and estradiol levels in FF were inversely related to age. Furthermore, FF inhibin B levels were inversely associated with serum FSH levels on day 3 of the menstrual cycle, which was believed to reflect the ovarian reserve. CONCLUSION: Inhibin B in FF may serve as an effective marker of follicular development and a useful predictor of quality of embryo. In addition, quality of oocyte is age related and declines as age increases. PMID- 12093831 TI - Clinical, endocrine and metabolic effects of metformin added to ethinyl estradiol cyproterone acetate in non-obese women with polycystic ovarian syndrome: a randomized controlled study. AB - BACKGROUND: Oral contraceptive pills (OC) are usually the first choice of treatment for polycystic ovarian syndrome (PCOS), when fertility is not desired. However, they do not improve, or may even further induce impairment of insulin sensitivity, which is already impaired in women with PCOS. In this prospective, randomized study, we analysed the additional benefits of adding metformin to the OC treatment in non-obese women with PCOS. METHODS: After a baseline work-up including body mass index (BMI), waist:hip ratio (WHR), Ferriman-Gallwey score, ovarian volume, serum gonadotrophin, androgen and sex hormone-binding globulin (SHBG) levels, and fasting lipid, glucose and insulin levels, 40 non-obese women with PCOS were assigned either to the OC or to the OC + metformin treatment by computer-assisted randomization. At the end of the 4 month follow-up period, subjects were re-evaluated. RESULTS: The two groups were similar at baseline. After treatment, women in the OC + metformin group had significant decreases in BMI and WHR, and a significant increase in insulin sensitivity, in contrast to those in the OC group, who had insignificant changes in these parameters. Adding metformin also caused significant improvements in serum androstenedione and SHBG levels compared with the OC treatment alone. CONCLUSIONS: Adding metformin to the OC treatment may improve the insulin sensitivity, and may further suppress the hyperandrogenaemia in non-obese women with PCOS. PMID- 12093832 TI - Pilot study on the use of sublingual misoprostol with mifepristone in termination of first trimester pregnancy up to 9 weeks gestation. AB - BACKGROUND: A combination of mifepristone and misoprostol provides an effective method of medical abortion for early pregnancy. A new route of administration of misoprostol, the sublingual route, was investigated in this study. METHODS: One hundred women who requested legal termination of pregnancy up to 63 days were given 200 mg of oral mifepristone followed 48 h later by 800 microg (4 x 200 microg tablets) of sublingual misoprostol. RESULTS: Ninety-four women (94%) had a complete abortion with this regimen. There was one ongoing pregnancy. The median duration of vaginal bleeding was 15 days. There were no serious complications. However, lower abdominal pain, diarrhoea, chills and fever were the commonest side-effects with incidences of 89, 42, 38 and 79% respectively. CONCLUSIONS: The combination of mifepristone and sublingual misoprostol is effective for medical abortion up to 63 days gestation. Randomized trials are required to compare its efficacy and side-effect profile with vaginal misoprostol. PMID- 12093833 TI - Mutation analysis of the inhibin alpha gene in a cohort of Italian women affected by ovarian failure. AB - BACKGROUND: Premature ovarian failure (POF) is a secondary hypergonadotrophic amenorrhoea affecting 1-3% of females, whose aetiology is almost unknown. However, inhibin alpha gene (INHalpha) has recently been indicated as candidate in POF pathogenesis. METHODS: We analysed patients affected by POF (n = 157) for the missense mutation (769G-->A transition) in the exon 2 of the INHalpha gene. The same analysis was carried out on early menopause (EM) (n = 36) and primary amenorrhoea (n = 12) patients. RESULTS: The incidence of the mutation was significantly more frequent within both POF (7/157, 4.5%) (Fisher's exact test, P = 0.030) and primary amenorrhoea (3/12, 25%) (Fisher's exact test, P < 0.001) patients, compared with the control population of women (0/100), who experienced physiological menopause. No mutation was found in EM patients. Furthermore, the likelihood of finding the mutation was statistically significant in familial (5/65; 7.7%) (Fisher's exact test, P < 0.01) but not in sporadic (2/92; 2.2%) (Fisher's exact test, P = not significant) POF, compared with the control group. The analysis of pedigrees showing the inheritance of the 769G-->A mutation and POF strengthens the concept of the disease heterogeneity, since the POF phenotype was not always associated with the mutation. Moreover, a higher prevalence of the C allele of a single nucleotide polymorphism (129C-->T), located in the 5'-UTR of the INHalpha gene, was observed in POF patients (80.3%) than in the control group (66.7%) (Fisher's exact test, P = 0.014). CONCLUSION: These data strengthen the concept of the INHalpha gene as a candidate for ovarian failure. PMID- 12093834 TI - A prospective study of the effects of female and male caffeine consumption on the reproductive endpoints of IVF and gamete intra-Fallopian transfer. AB - BACKGROUND: This study evaluated the timing and amount of caffeine intake by women and men undergoing IVF and gamete intra-Fallopian transfer (GIFT) on oocyte retrieval, sperm parameters, fertilization, multiple gestations, miscarriage, and live births. METHODS: A prospective study of 221 couples was conducted in Southern California between 1993 and 1998. "Usual" caffeine intake during lifetime and 1 year prior to attempt, caffeine intake during the week of the initial clinic visit, as well as intake during the week of the procedure, was evaluated from beverages (coffee, soda, tea) and chocolates. RESULTS: Not achieving a live birth was significantly associated with "usual" female caffeine consumption [adjusted odds ratios (95% confidence intervals): 3.1 (1.1, 9.7) and 3.9 (1.3, 11.6) for intake of >2-50 and 50 mg/day, compared with 0-2 mg/day] and consumption during the week of the initial visit [2.9 (1.1, 7.5) and 3.8 (1.4, 10.7)] female compared with 0-2 mg/day, although caffeine use was low. Infant gestational age decreased by 3.8 (-6.9, -0.7) or 3.5 (-6.7, -0.3) weeks for women who consumed >50 mg/day of caffeine "usually" or during the week of the initial visit. The odds of having multiple gestations increased by 2.2 (1.1, 4.4) and 3.0 (1.2, 7.4) for men who increased their "usual" intake or intake during the week of the initial visit by an extra 100 mg/day. Caffeine intake was not significantly associated with other outcomes. CONCLUSIONS: This is the first IVF/GIFT study to report any effect of caffeine on live births, gestational age, and multiple gestations. If these findings are replicated, caffeine use should be minimized prior to and while undergoing IVF/GIFT. PMID- 12093835 TI - Obstetric outcome among women with unexplained infertility after IVF: a matched case-control study. AB - BACKGROUND: Infertility itself and also assisted reproductive treatment increase the incidence of some obstetric complications. Women with unexplained infertility are reported to be at an increased risk of intrauterine growth restriction during pregnancy, but not for other perinatal complications. METHODS: A matched case control study was performed on care during pregnancy and delivery, obstetric complications and infant perinatal outcomes of 107 women with unexplained infertility, with 118 clinical pregnancies after IVF or ICSI treatment. These resulted in 90 deliveries; of these, 69 were singleton, 20 twin and one triplet. Two control groups were chosen from the Finnish Medical Birth Register, one group for spontaneous pregnancies (including 445 women and 545 children), matched according to maternal age, parity, year of birth, mother's residence and number of children at birth, and the other group for all pregnancies after IVF, ICSI or frozen embryo transfer treatment (FET) during the study period (including 2377 women and 2853 children). RESULTS: Among singletons, no difference was found in the mean birthweight, and the incidence of low birthweight (<2500 g) was comparable with that of the control groups. No differences were found in gestational duration, major congenital malformations or perinatal mortality among the groups studied. Among singletons in the study group, there were more term breech presentations (10.1%) compared with both spontaneously conceiving women and all IVF women (P < 0.01). The rate of pregnancy-induced hypertension was significantly lower among singletons in the study group (P < 0.05) compared with other IVF singletons. The multiple pregnancy rate was 23.3% in the study group. The obstetric outcome of the IVF twins was similar to both control groups. CONCLUSIONS: The overall obstetric outcome among couples with unexplained infertility treated with IVF was good, with similar outcome compared with spontaneous pregnancies and IVF pregnancies generally. PMID- 12093836 TI - Ultrasound-guided embryo transfer improves pregnancy rates and increases the frequency of easy transfers. AB - BACKGROUND: Recent reports have suggested that ultrasound (US) guidance during embryo transfer might improve pregnancy rates. METHODS: A prospective randomized (computer-generated random table) trial was performed to compare embryo transfer under abdominal US guidance (n = 255 women) with clinical touch embryo transfer (n = 260). RESULTS: The clinical pregnancy rate was 26.3% (67/255) in the US guided transfer group compared with 18.1% (47/260) in the clinical touch transfer group (P < 0.05). The implantation rate was 11.1% (100/903) in the US group compared with 7.5% (66/884) in the clinical touch group (P < 0.05). US-guided transfer was associated with a decrease in the difficulty of the transfers: 97% of transfers were easy in the US-guided group compared with 81% in the clinical touch group (P < 0.05). CONCLUSIONS: US-guided embryo transfer increased pregnancy and implantation rates in IVF cycles, as well as the frequency of easy transfers. It is suggested that the decrease in cervical and uterine trauma can play a role in the increase in pregnancy rates associated with US-guided transfer. It is recommended that embryo transfer should be performed under US guidance. PMID- 12093837 TI - Ultrasound measurement of the uterocervical angle before embryo transfer: a prospective controlled study. AB - BACKGROUND: The study aim was to determine whether moulding the embryo transfer catheter according to the uterocervical angle measured by ultrasound could improve pregnancy and implantation rates. METHODS: Patients were alternately allocated to one of two groups. In the ultrasound-guided group (n = 320), the catheter was moulded according to the uterocervical angle measured by abdominal ultrasound. In controls (n = 320), embryo transfer was performed using the "clinical feel" method. RESULTS: Moulding the embryo transfer catheter according to the uterocervical angle significantly increased clinical pregnancy [(OR = 1.57, 95% CI (1.08-2.27)] and implantation rates [(OR = 1.47, 95% CI (1.10-1.96)] compared with the "clinical feel" method. It also significantly reduced difficult transfers [(OR = 0.25, 95% CI (0.16-0.40)] and blood during transfers [OR = 0.71, 95% CI (0.50-0.99)]. Patients with large angles (>60 degrees ) had significantly lower pregnancy rates compared with those with no angle [OR = 0.36, 95% CI (0.16 0.52)]. CONCLUSIONS: Moulding the embryo transfer catheter according to the uterocervical angle measured by ultrasound increases clinical pregnancy and implantation rates and diminishes the incidence of difficult and bloody transfers. PMID- 12093838 TI - Effects of hepatitis C virus on the apoptosis percentage of granulosa cells in vivo in women undergoing IVF: preliminary results. AB - BACKGROUND: The aim of this study was to investigate the relationship between the apoptosis percentage of human luteinized granulosa cells (GC) and the presence of hepatitis C virus (HCV) in follicular fluid (FF). METHODS: GC were isolated from FF of 12 women undergoing 12 IVF cycles: six were HCV+ with active viral replication and six HCV- serving as controls. No male partner was HCV+. HCV detection and quantification were assessed by reverse transcriptase-polymerase chain reaction in serum, FF and embryo-incubation medium. GC were analysed by flow cytometry after propidium iodide staining to measure the percentages of apoptotic GC. Routine IVF parameters were tabulated. RESULTS: Mean +/- standard deviation (SD) serum and FF HCV viral loads were 3.58 +/- 4.25 x 10(6) and 0.14 +/- 0.10 x 10(6) IU/ml respectively. Mean percentages of apoptotic GC from HCV+ and HCV- women were 3.08 +/- 1.14 and 3.14 +/- 1.40% respectively. No statistically significant difference was found between these two groups concerning GC apoptosis and when we compared all IVF parameters. No HCV RNA was detected in embryo incubation media after 2 days of culture. CONCLUSIONS: Comparing GC apoptosis percentages and usual IVF parameters in the HCV+ group versus the HCV- group, our preliminary study shows that active chronic HCV infection does not affect follicle development and IVF outcome in HCV+ women undergoing IVF. Furthermore, the risk of newborns becoming HCV-infected might not be increased by assisted reproductive technologies when performed in couples in which women are HCV+ and men HCV-. PMID- 12093840 TI - Primordial and pre-antral follicles are not commonly observed in IVF aspirates. AB - BACKGROUND: Follicular fluid recovered from IVF patients has been proposed to be a valuable source of pre-antral and primary follicles for patient therapy and research. We evaluated the recovery of immature follicles in follicular fluid from 54 patients undergoing IVF using several techniques. METHODS: Fluid from each patient underwent several methods of follicle recovery including: filtration through a cell strainer, Ficoll-Paque density gradient, isolate density gradient, histological slide preparation, and enzymatic digestion with collagenase and DNase. RESULTS: 34 primordial and primary follicles, mean 0.63 +/- 0.27/patient, and 14 pre-antral follicles, mean 0.26 +/- 0.14/patient, were found in this study. The serum estradiol level on the day of HCG injection was significantly lower (P < 0.05) in patients in which immature follicles were recovered, compared with those without immature follicles in the follicular fluid (1779.9 +/- 167.6 versus 2246.6 +/- 153.2 pg/ml). There were no women with advanced maternal age (>39 years) who had immature follicles in the follicular fluid. CONCLUSIONS: Follicular fluid cannot be considered an efficient or reliable source of immature follicles. The presence of any immature follicles appears to be associated with cause of infertility, the random placement of the aspirating needle and may be related to the age of patient. PMID- 12093839 TI - Oocyte and embryo quality after coasting: the experience from oocyte donation. AB - BACKGROUND: Oocyte donation provides us with an opportunity to study the clinical outcome of oocytes, retrieved from women undergoing coasting, in recipients in whom endometrial receptivity is unaltered by the coasting procedure. Thus, our aim was to describe oocyte donation outcome in donors undergoing coasting, the oocyte and embryo quality obtained from these cycles, and to determine the influence of coasting duration in the cycle outcome. METHODS: Matched-paired analysis included 15 oocyte donors with high response to ovarian stimulation and submitted to coasting and 15 oocyte donors with normal response to ovarian stimulation and not undergoing coasting. There were 38 oocyte recipients who shared oocytes from the donors under coasting and 37 from donors not undergoing coasting. RESULTS: Both groups of donors were comparable in terms of days and dose of ovarian stimulation, oocytes retrieved, metaphase II oocytes obtained, and in the appearance of ovarian hyperstimulation syndrome. Both groups of oocyte recipients were comparable in male-associated factor, pregnancy and implantation rates, as well as in embryo quality. Recipients from donors with coasting for >4 days had significantly lower implantation and pregnancy rates. CONCLUSIONS: the outcome of oocyte donation from donors undergoing coasting is not impaired, as good implantation and pregnancy rates are achieved. Embryo quality, according to our current standards, does not seem to be compromised by coasting itself. However, if coasting in oocyte donors is prolonged for >4 days there is a significant decrease in both implantation and pregnancy rates. PMID- 12093841 TI - Retrospective multicentre study on mechanical and enzymatic preparation of fresh and cryopreserved testicular biopsies. AB - BACKGROUND: Isolation of sperm suitable for ICSI from fresh or frozen-thawed testicular sperm extraction (TESE) can be facilitated by mechanical or enzymatic processing of the samples. METHODS: A retrospective multicentre study was initiated to compare these two approaches. Eleven German centres provided data on their TESE cycles performed during the period 1996/1997. Quality of retrieved sperm, fertilization rates of injected oocytes, embryo quality, resulting pregnancy rates and evolution of pregnancies were evaluated. RESULTS: The percentage of cycles with at least some motile sperm available for injection was higher after mechanical preparation. Independent of the preparation method, fertilization rates were higher for motile compared with immotile sperm or elongated spermatids in all groups and in general higher for cryopreserved versus fresh samples. Embryo quality was significantly better after injection of motile sperm for all treatments and in particular after enzymatic versus mechanical processing of biopsies. Pregnancy rates were identical for embryos derived from sperm prepared mechanically or enzymatically from fresh or cryopreserved testicular samples. The abortion rate (32/172, 18.6%) and the rate of multiple implantations (32/140, 22.9%) were not different from results reported in the literature for ICSI using ejaculated sperm. CONCLUSION: In this retrospective multicentre study, no unequivocal advantage of one over the other preparation method could be identified in 839 ICSI cycles using testicular sperm from 549 patients. PMID- 12093842 TI - Impact of repeated testicular fine needle aspirations (TEFNA) and testicular sperm extraction (TESE) on the microscopic morphology of the testis: an animal model. AB - BACKGROUND: The aim of this work was to compare testicular histological changes occurring following testicular fine needle aspiration (TEFNA) and testicular sperm extraction (TESE). METHODS: TESE or TEFNA were performed on normal male rats, in a similar manner to azoospermic men. The animals were killed after 7, 14 and 31 days and their testes were examined. RESULTS: TESE caused chronic inflammation, occasional necrosis and degenerative changes in testicular germ cells in only approximately 10% of the remaining testicular tissue. TEFNA caused widespread architectural distortion of seminiferous tubules into irregular and deformed lumens lined by Sertoli cells only, as well as focal chronic inflammation, necrosis and degenerative changes accompanied by decreased spermatogenesis. Similar but less extensive changes were noted when fewer punctures were performed. When negative suction pressure was not applied during TEFNA, similar histological changes occurred, indicating that testicular damage was related to the puncture itself. Following either procedure, the contralateral non-operated testes were unaffected. CONCLUSION: In this animal model, TEFNA inflicts severe, progressive and irreversible damage on the architecture of the tubules in the needle's path. The multi-focal nature of this technique eventually leads to widespread tubular atrophy that is proportional to its extent. In contrast, TESE causes localized scarring and fibrosis, rendering most of the remaining testicle intact. The clinical relevance of such findings, produced in normal animal testes, to testes of azoospermic men, is yet to be determined. PMID- 12093843 TI - Predictive value of testicular histology in secretory azoospermic subgroups and clinical outcome after microinjection of fresh and frozen-thawed sperm and spermatids. AB - BACKGROUND: A retrospective study was carried out on 159 treatment cycles in 148 secretory azoospermic patients to determine whether histopathological secretory azoospermic subgroups were predictive for gamete retrieval, and to evaluate outcome of microinjection using fresh or frozen-thawed testicular sperm and spermatids. METHODS: Sperm and spermatids were recovered by open testicular biopsy and microinjected into oocytes. Fertilization and pregnancy rates were assessed. RESULTS: In hypoplasia, 97.7% of the 44 patients had late spermatids/sperm recovered. In maturation-arrest (MA; 47 patients), 31.9% had complete MA, and 68.1% incomplete MA due to a focus of early (36.2%) or late (31.9%) spermiogenesis. Gamete retrieval was achieved in 53.3, 41.2 and 93.3% of the cases respectively. In Sertoli cell-only syndrome (SCOS; 57 patients), 61.4% were complete SCOS, whereas incomplete SCOS cases showed one focus of MA (5.3%), or of early (29.8%) and late (3.5%) spermiogenesis. Only 29.8% of the patients had a successful gamete retrieval, 2.9% in complete and 77.3% in incomplete SCOS cases. In total, there were 87 ICSI, 39 elongated spermatid injection (ELSI) and 33 round spermatid injection (ROSI) treatment cycles, with mean values of fertilization rate of 71.4, 53.6 and 17%, and clinical pregnancy rates of 31.7, 26.3 and 0% respectively. CONCLUSIONS: Histopathological subgroups were positively correlated with successful gamete retrieval. No major outcome differences were observed between testicular sperm and elongated spermatids, either fresh or frozen-thawed. However, injection of intact round-spermatids showed very low rates of fertilization and no pregnancies. PMID- 12093845 TI - Sperm DNA damage in potentially fertile homozygous beta-thalassaemia patients with iron overload. AB - BACKGROUND: To test the hypothesis that human sperm DNA could sustain iron induced oxidative damage and reduce its fertilizing ability, we studied patients with homozygous beta-thalassaemia major (HbTh) as a model of iron overload. METHODS: Sperm from six thalassaemic patients and five age-matched controls were assessed by the sperm chromatin structure assay (SCSA) and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) assay. Semen parameters, endocrine markers of testicular function, iron profiles and the presence of organ dysfunction were also determined. RESULTS: All patients with HbTh were iron overloaded (median ferritin: 2251 microg/l) and had evidence of spontaneous spermatogenesis. Thalassaemic patients had more sperm DNA damage than the controls (P < 0.01). The sperm DNA damage by SCSA and TUNEL were positively correlated (P < 0.05). Sperm motility and TUNEL results were negatively correlated (P < 0.05), while the age of onset of chelation and sperm DNA damage were positively associated with both SCSA (R(2) = 0.80, P = 0.016) and TUNEL data (R(2) = 0.67, P < 0.044). No other biochemical or clinical data were associated with sperm DNA damage. CONCLUSIONS: The increase in sperm DNA damage and the negative correlation between sperm motility and DNA damage suggest that iron overload in HbTh predisposes sperm to oxidative injury. This finding has important implications in assisted reproductive procedures such as ICSI where there is increased risk of transmitting defective DNA to the offspring. PMID- 12093844 TI - In-vitro effects of FSH and testosterone withdrawal on caspase activation and DNA fragmentation in different cell types of human seminiferous epithelium. AB - BACKGROUND: Caspases are downstream elements of apoptosis-mediating pathways initiated by the Fas ligand/Fas receptor system which is supposed to play a central role in the regulation of apoptosis in the human seminiferous epithelium. However, caspase activity in different cell types of this epithelium has never been addressed. METHODS AND RESULTS: We evaluated caspase activity and DNA integrity in Sertoli and germ cells within in-vitro cultured segments of human seminiferous tubules after induction of apoptosis by FSH or testosterone withdrawal. FSH withdrawal increased the incidence of DNA fragmentation in meiotic (primary spermatocytes) and post-meiotic (spermatids) germ cells without producing any detectable effect on caspase activity in these cells and without affecting DNA integrity or caspase activity in Sertoli cells. Testosterone withdrawal stimulated caspase activity and produced DNA fragmentation in Sertoli cells, but showed only a weak effect on DNA fragmentation in germ cells and did not alter germ cell caspase activity. CONCLUSIONS: These findings confirm the central role of caspases in apoptosis of Sertoli cells. However, they also suggest that acute apoptosis of germ cells in the adult human testis occurs in a caspase-independent way and is controlled by Sertoli cells via an as yet undetermined mechanism. PMID- 12093846 TI - Sperm aneuploidy rates in younger and older men. AB - BACKGROUND: In order to assess the possible risk of chromosomal abnormalities in offspring from older fathers, we investigated the effects of age on the frequency of chromosomal aneuploidy rates of human sperm. METHODS AND RESULTS: Semen samples were collected from 15 men aged <30 years (24.8 +/- 2.4 years) and from eight men aged >60 years (65.3 +/- 3.9 years) from the general population. No significant differences in ejaculate volume, sperm concentration and sperm morphology were found, whereas sperm motility was significantly lower in older men (P = 0.002). For the hormone values, only FSH was significantly elevated in the older men (P = 0.004). Multicolour fluorescence in-situ hybridization was used to determine the aneuploidy frequencies of two autosomes (9 and 18); and of both sex chromosomes using directly labelled satellite DNA probes on decondensed sperm nuclei. A minimum of 8000 sperm per donor and >330 000 sperm in total were evaluated. The disomy rates per analysed chromosomes were 0.1-2.3% in younger men and 0.1-1.8% in older men. The aneuploidy rate determined for both sex chromosomes and for the autosomes 9 and 18 were not significantly different between the age groups. CONCLUSIONS: The results suggest that men of advanced age still wanting to become fathers do not have a significantly higher risk of procreating offspring with chromosomal abnormalities compared with younger men. PMID- 12093847 TI - Achievement of pregnancy in globozoospermia with Y chromosome microdeletion after ICSI. AB - Pregnancy achieved with sperm from a patient with globozoospermia is rare, even after ICSI, since the activation of the oocyte may not occur in this disorder. Therefore, activation of the oocytes by piezoelectricity or calcium ionophores has been suggested, although spontaneous activation of the oocyte after ICSI has been reported in some cases. We report a successful pregnancy in a couple in which the male partner had globozoospermia with microdeletions in the Y chromosome with no further assisted activation after ICSI. During the diagnostic study of the husband, increased numerical chromosome abnormalities after fluorescent in-situ hybridization (FISH) and microdeletions in AZFa; sY86 and AZFb; sY 131 were detected. Out of the 13 oocytes injected, four fertilized and a twin pregnancy was obtained after replacement of four embryos. Healthy twin girls were delivered after a term pregnancy. Some patients with globozoospermia may also have Y chromosome microdeletions, which subsequently may be inherited by the male offspring in cases of achievement of pregnancy. PMID- 12093848 TI - Correlation of scrotal temperature in twins. AB - BACKGROUND: Scrotal temperature and semen quality have been found to be inversely correlated in several studies, and variation in scrotal temperature may contribute to the well known variation in semen quality. The reason for the variation in scrotal temperature is not well understood although determinants could be of a genetic nature. METHODS: We monitored scrotal skin temperature for 24 h in a group of monozygotic and dizygotic twins and single-born brothers (n = 48 males). A thermistor was attached to the underwear and the temperature of the scrotal skin was recorded every 5 min using a small portable data logger. RESULTS: A correlation in median scrotal temperature was found among monozygotic twins (r = 0.64, P = 0.01), but not in dizygotic twins and single-born brothers (r = 0.17, NS). Similar results were found for other percentiles of temperature. CONCLUSIONS: The results suggest a genetic component to the variation in scrotal temperature. An hereditary element in male fecundity may be expressed through scrotal temperature, which constitutes a mechanism independent of those responsible for the development of the sperm producing epithelium. PMID- 12093849 TI - Trophectoderm growth and bilateral symmetry of the blastocyst in the mouse. AB - BACKGROUND: The present study was undertaken to ascertain whether the polarized flow of cells from polar to mural trophectoderm is related to the axis of bilateral symmetry of the blastocyst in the mouse, and whether trophectoderm cells can initiate new cycles once they have left the polar region. METHODS AND RESULTS: Two different approaches were used to investigate the relationship of polar to mural flow of trophectoderm cells to the bilateral axis. One was to mark peripheral polar trophectoderm cells at one or both ends of the bilateral axis in early blastocysts and examine the distribution of their clonal descendants after further growth in culture. The other was to mark the two ends of the bilateral axis with small oil drops in the zona pellucida in blastocysts whose polar trophectoderm was then labelled globally with fluorescent latex microspheres before culture. In both cases, marking of additional blastocysts orthogonal to the bilateral axis was also done. The results show that the direction of polar to mural flow of cells is not random, and that the most distal mural trophectoderm cell could yield up to eight descendants during 45 h of culture. CONCLUSION: The findings are consistent with the polar to mural flow of trophectoderm cells being aligned with the bilateral axis. Moreover, trophectoderm cells can embark on new cycles even when remote from the inner cell mass. PMID- 12093850 TI - A prospective trial of blastocyst culture and transfer. AB - BACKGROUND: The aim of this study was to evaluate the effectiveness of the blastocyst culture method compared with the conventional day 3 transfer method using a prospective trial. METHODS: A total of 235 patients with 273 cycles were evaluated for a period of almost 1 year. Depending upon the sequence in which the ovum retrieval was performed, patients were prospectively assigned (alternate allocation) to a culture period of 3 or 5 days duration. Embryos were transferred either on day 3 (after culture in human tubal fluid) or on day 5 (after culture in sequential media). RESULTS: The pregnancy rates after embryo transfer on days 3 and 5 were similar at 26.5 and 25.9% respectively. Among the day 5 embryo transfer group, patients were divided into three groups corresponding to three sequential media. The pregnancy rates were 32.0% using Irvine blastocyst medium, 6.9% using G1.2/G2.2 and 32.4% using Cook blastocyst medium. CONCLUSIONS: The results of our study were not as successful as previous studies had been. Additionally, there may have been problems in day 5 embryo transfer, such as choosing the sequential media, quality control, contamination and so on. From the results of this study, it appears that day 5 embryo transfer has no advantages for ordinary patients of assisted reproductive technology. PMID- 12093851 TI - Day 3 embryo transfer with combined evaluation at the pronuclear and cleavage stages compares favourably with day 5 blastocyst transfer. AB - BACKGROUND: The respective advantages of day 3 and day 5 embryo transfer are a matter of debate. Previous comparisons did not include pronuclear stage zygote scoring and cumulative success rates (fresh and cryopreserved embryos). METHODS: Patients were randomized prospectively for day 3 or day 5 embryo transfer. Day 3 embryos were selected for transfer and cryopreservation by using combined evaluation at the pronuclear and cleavage stages. RESULTS: There was no difference between day 3 and day 5 fresh embryo transfers as to the rates of pregnancy (58 versus 62%), clinical pregnancy (56 versus 58%), delivery (50 versus 48%), implantation (35 versus 38%) and birth (33 versus 36%) rates. The corresponding values for cryopreserved embryo transfers were also similar. However, day 3 embryo transfer compared favourably with day 5 transfer when the pregnancy (90 versus 66%), clinical pregnancy (85 versus 62%) and delivery (77 versus 52%) rates were calculated per oocyte recovery attempt. CONCLUSIONS: With a selected population of good prognosis patients and our embryo selection criteria, the implantation potential of day 3 and day 5 embryos is equal. Per oocyte recovery attempt, day 3 transfer is more clinically efficient than day 5 transfer, but at least one transfer of cryopreserved embryos is necessary to manifest this superiority. PMID- 12093852 TI - Embryo quality and IVF treatment outcomes may correlate with different sperm comet assay parameters. AB - BACKGROUND: Standard semen parameters have proven poor at predicting the outcomes of IVF treatment cycles. As recent studies suggest that the male genome may play an important role in early embryogenesis, this study attempts to correlate the level of sperm DNA damage in fresh semen and prepared sperm with the outcomes of conventional IVF treatment cycles. METHODS: Forty patients embarking on IVF treatment were recruited into this prospective observational study. Both fresh semen and PureSperm-prepared sperm were processed using a modified comet assay 3 6 months prior to the patients' IVF treatment cycles. Comet head DNA (mean and integrated head density) and tail DNA parameters (length and moment) were measured separately. RESULTS: Significant correlations between total sperm concentration and between comet length, moment, mean head density with embryo quality were detected in fresh semen and prepared sperm. Surprisingly, no significant correlations between head and tail parameters were detected. CONCLUSIONS: Comet head and tail DNA parameters appear to be potentially useful as predictors of embryo quality and IVF outcomes, especially in couples with unexplained subfertility. The lack of correlation between head and tail parameters may be due to a different mechanism of DNA damage within these two compartments. PMID- 12093853 TI - Morphological appearance of the cryopreserved mouse blastocyst as a tool to identify the type of cryoinjury. AB - BACKGROUND: If it were possible to deduce the mechanism of injury in cryopreserved embryos by their appearance, it would help to optimize cryopreservation protocols. METHODS: Mouse blastocysts were treated so that they were damaged by the six types of cryoinjuries listed below, and their appearance was observed at recovery in sucrose solution and a modified phosphate-buffered saline (PB1), and after culture for 1 and 24 h. RESULTS: (i) Intracellular ice: the embryos shrank normally in sucrose solution, but swelled in PB1 and collapsed after culture. (ii) Chemical toxicity of the cryoprotectant: the embryos looked normal in sucrose solution and PB1. After 1 h of culture, however, the blastomeres showed decompaction and degenerated thereafter. If the toxicity was extremely high, embryos looked nearly normal in PB1, but the surface of the cytoplasm was wrinkled as if they were "fixed". (iii) Osmotic swelling: the embryos looked normal in PB1, but after culture they shrank. (iv) Osmotic shrinkage: the embryos swelled in PB1, and then collapsed. (v) Fracture damage: the zona pellucida of the embryos was dissected. (vi) Extracellular ice: the zona of the embryos was elongated. CONCLUSIONS: It was often possible to deduce the type of injury that had occurred in cryopreserved embryos from their appearance at recovery and during subsequent culture. This may help to improve cryopreservation protocols for embryos of many species, including man. PMID- 12093854 TI - Permeability characteristics and osmotic sensitivity of rhesus monkey (Macaca mulatta) oocytes. AB - BACKGROUND: Permeability characteristics and sensitivity to osmotic shock are principal parameters that are important to derive procedures for the successful cryopreservation of mammalian oocytes. METHODS AND RESULTS: The osmotically inactive volume of rhesus monkey oocytes was determined by measuring their volumes in the presence of hypertonic solutions of sucrose from 0.2 to 1.5 mol/l, compared with their volume in isotonic TALP-HEPES solution. Boyle-van't Hoff plots at infinite osmolality indicated that the non-osmotic volumes of immature and mature oocytes were 20 and 17% respectively. Osmotic responses of oocytes exposed to 1.0 mol/l solutions of glycerol, dimethylsulphoxide (DMSO) and ethylene glycol (EG) were determined. Rhesus monkey oocytes appeared to be less permeable to glycerol than to DMSO or to EG. Sensitivity of oocytes to osmotic shock was determined by exposing them to various solutions of EG (0.1 to 5.0 mol/l) and then abruptly diluting them into isotonic medium. Morphological survival, as measured by membrane integrity, of oocytes diluted out of EG depended significantly on the concentration of EG (P < 0.01). CONCLUSION: Determination of permeability characteristics and sensitivity to osmotic shock of rhesus oocytes will aid in the derivation of procedures for their cryopreservation. PMID- 12093855 TI - A confocal microscopy analysis of the spindle and chromosome configurations of human oocytes cryopreserved at the germinal vesicle and metaphase II stage. AB - BACKGROUND: Routine oocyte cryopreservation remains an elusive technique in the wide range of assisted reproductive technologies available. This study examines the effect of a cryopreservation protocol on the spindle and chromosome configurations of human oocytes cryopreserved at the germinal vesicle (GV) and metaphase II (MII) stage. METHODS: GV oocytes were randomly assigned to one of three groups: (i) control oocytes matured in vitro to MII stage (n = 156); (ii) oocytes cryopreserved at the GV stage and then matured in vitro (n = 90); (iii) oocytes cryopreserved at the MII stage (n = 147). Following cryopreservation and in-vitro maturation, immunostaining of tubulin and chromatin was performed, before visualization using confocal microscopy. RESULTS: A statistically significant increase was observed in the survival rate in group 2 (73.3%, 66/90) compared to group 3 (55.7%, 82/147) (P < 0.007). Exposure of oocytes to the cryoprotective solutions without freezing had no effect on the structure of their second meiotic spindle. However, statistically significant differences were observed on both spindle and chromosome configurations of oocytes from group 2 (5.2 and 5.2% respectively) and group 3 (16.2 and 18.8% respectively) compared with group 1 oocytes (71.6 and 82.0% respectively) (P < 0.001 in all cases). CONCLUSIONS: The protocol followed results in high rates of survival and potential for in-vitro maturation, but has a deleterious effect on the organization of the meiotic spindle of human oocytes cryopreserved at both the GV and MII stages. PMID- 12093856 TI - A human oocyte with two sets of MII/PB-structures. AB - This paper reports a case where a normal-sized human oocyte has been documented having two polar bodies and two metaphase spindles. This finding suggests that the chromosomal content in oocytes of normal size may occasionally be duplicated and offers an additional explanation for the origin of triploid zygotes in humans. We speculate that this duplication may be caused by oocyte fusion at an early stage. PMID- 12093857 TI - Hyperalgesia, nerve infiltration and nerve growth factor expression in deep adenomyotic nodules, peritoneal and ovarian endometriosis. AB - BACKGROUND: The aim of this study was to investigate a possible role for nerve growth factor (NGF) in the mechanism of pain and hyperalgesia induced by deep adenomyotic nodules and other forms of endometriosis and to clarify the relationship between endometriotic lesions and the surrounding nerves. METHODS: Endometriotic lesions (deep adenomyotic nodules, peritoneal endometriosis, ovarian endometriosis) and eutopic endometrium were obtained from 51 patients presenting with pain. Patients were allocated to two groups (group 1: patients with a deep adenomyotic nodule (n = 23); group 2: patients with peritoneal and/or ovarian endometriosis but without deep adenomyotic nodule (n = 28). Immunohistochemistry with antibodies against NGF, NGF specific tyrosine-kinase receptor (Trk-A) and S-100 protein was performed. Results were expressed as mean H-scores +/- SD, and correlated with the presence of hyperalgesia. RESULTS: The percentage of patients presenting hyperalgesia at physical examination was significantly higher in group 1 (96%) than in group 2 (11%) (P < 0.001). NGF expression was significantly stronger in deep adenomyotic nodules (DAN) than in ovarian (OE) and peritoneal endometriosis (PE), both in the proliferative phase in the glands [DAN: 226 +/- 18; OE: 140 +/- 9 (P < 0.001); PE: 110 +/- 7 (P < 0.001)] and in the stroma [(DAN: 204 +/- 21; OE: 125 +/- 15 (P < 0.001); PE: 100 +/- 9 (P < 0.01)]. NGF expression in DAN is also significantly stronger than in OE and PE in the secretory phase in the glands [DAN:181 +/- 32; OE: 85 +/- 3.3 (P < 0.001); PE: 65 +/- 9 (P < 0.001)] and in the stroma [DAN: 173 +/- 28; OE: 85 +/ 3.7 (P < 0.001); PE: 35 +/- 13 (P < 0.001)]. Perineurial and intraneurial invasion by endometriotic lesions were found only in deep adenomyotic nodules and not in the other forms of endometriosis. The specific receptor for NGF (Trk-A) is expressed in all the nerves that were included in the biopsies. CONCLUSIONS: These results suggest a role of NGF in endometriotic pain and hyperalgesia in deep adenomyotic nodules. The strong expression of the NGF-TrkA pathway in deep adenomyotic nodules could explain why this type of lesion infiltrates in richly innervated anatomical sites. PMID- 12093858 TI - Serum HCG 12 days after embryo transfer in predicting pregnancy outcome. AB - BACKGROUND: Assisted reproduction treatment (ART) entails a risk of ectopic pregnancy and early pregnancy loss. Serum HCG has been found to be predictive of pregnancy outcome. Our aim was to assess the clinical value of a single early HCG assay in ART pregnancies taking into account the aetiology and treatment of infertility. METHODS: During 1994-1999, we studied 774 embryo transfer cycles resulting in pregnancy defined as a serum HCG concentration of > or =5 IU/l on day 12 following embryo transfer. The treatment included IVF in 518, ICSI in 119, and frozen embryo transfer in 137 cycles. Serum HCG concentrations were measured by fluoroimmunometric assay. Pregnancies were classified as viable (live fetus at > or =22 weeks gestation) or non-viable (biochemical pregnancy, miscarriage, ectopic pregnancy and molar pregnancy). Data on the outcomes were retrospectively retrieved from the records. RESULTS: The median HCG concentration was 126 IU/l in viable pregnancies and 31 IU/l in non-viable pregnancies (P < 0.0001). The median HCG concentration was 115 IU/l in singleton pregnancies and 201 IU/l in multiple pregnancies (P < 0.0001). Male factor infertility was associated with viable pregnancies (P = 0.004) and tubal factor with non-viable pregnancies (P = 0.003); the lowest HCG level (88 IU/l) was observed in subjects with both male factor infertility and ICSI treatment (P = 0.001). An HCG value of 76 IU/l emerged as the most suitable cut-off point to predict viable pregnancy. Probabilities of each type of outcome related to the HCG level are given. CONCLUSIONS: A single HCG reading on day 12 after embryo transfer helps to plan the subsequent follow up. Male factor infertility and ICSI are associated with relatively low HCG values in viable pregnancies. PMID- 12093859 TI - Salivary progesterone levels and rate of ovulation are significantly lower in poorer than in better-off urban-dwelling Bolivian women. AB - BACKGROUND: Agriculturalists in less-developed countries (LDC) have lower progesterone levels than urban industrialized populations. However, it is unknown if urban LDC populations are also relatively lower. We tested whether urban Bolivia samples-poorer (Bol-p) and better-off (Bol-b)-have lower progesterone than a Chicago (USA) sample, and whether progesterone and rate of ovulation are lower in Bol-p than in Bol-b. METHODS: Serial salivary samples collected from Bolivians, screened according to strict exclusion criteria during two complete menstrual cycles, were radioimmunoassayed for progesterone; anthropometrics were collected at mid-follicular and mid-luteal phases. RESULTS: Progesterone levels are lower in the Bolivia samples, and higher in the Bol-b than Bol-p; ovulation rate is greater in Bol-b than Bol-p. For only ovulatory cycles, mean-follicular-P (pmol/l), mean-luteal-P (pmol/l), and mean-peak-P (pmol/l) are respectively 65, 142 and 208 in Bol-p; 76, 167 and 232 in Bol-b; and 96, 240 and 330 in Chicago. Principal components representing body-size and progesterone level are positively correlated (r = 0.404, P = 0.005). CONCLUSIONS: Progesterone levels appear to be influenced by chronic and acute ecological conditions, evidenced by the association with body-size and the probability of ovulation respectively. These findings have implications for understanding cancer aetiology, developing population-appropriate hormonal contraceptives, and modelling the evolution and functioning of the reproductive system. PMID- 12093860 TI - The relationship between Caesarean section and subfertility in a population-based sample of 14 541 pregnancies. AB - BACKGROUND: There has been a threefold increase in the rate of Caesarean section over the past 25 years. The long-term consequences of Caesarean section may include subsequent subfertility. METHODS: We investigated the relationship between Caesarean section and subfertility within a cohort of 14 541 pregnant women. RESULTS: A history of previous Caesarean section was associated with an increased risk of taking >1 year to conceive from the time of planning a pregnancy, adjusted odds ratio (OR) 1.53 [95% confidence interval (CI) 1.09, 2.14]. This association was stronger for women of parity > or =2, adjusted OR 2.97 (95% CI 1.72, 5.10). Nulliparous women with a history of subfertility were at increased risk of delivery by Caesarean section, adjusted OR 1.56 (1.22, 2.00) and OR 2.33 (1.64, 3.30) for durations of >1 and >3 years respectively. CONCLUSIONS: These findings suggest a complex relationship between Caesarean section and subfertility where subfertility may both precede and be a consequence of Caesarean section. PMID- 12093861 TI - Is estradiol cardioprotection a nitric oxide-mediated effect? AB - BACKGROUND: Estradiol exerts a number of biological effects that support extensive observational data suggesting a protective role for estrogen in cardiovascular disease prevention. These include effects on lipid and carbohydrate metabolism, coagulation/fibrinolysis as well as a possible effect on vascular reactivity. It has been proposed that this might be mediated by vascular endothelial nitric oxide (NO) production. Accordingly, we designed complementary in-vivo and in-vitro studies to investigate this hypothesis further. METHODS: Firstly, in a group of 10 healthy post-menopausal women, bilateral venous occlusion plethysmography was used to examine forearm vasoconstrictor responses to intrabrachial N(G)-monomethyl-l-arginine (l-NMMA; a substrate inhibitor of nitric oxide synthase) both before and after 4 weeks of treatment with transdermal 17beta-estradiol (E(2)) (80 microg/day). Secondly, we examined the direct effects of acute (24 h) and chronic (7 days) treatment with E(2) (10 pmol/l and 10 nmol/l) on endothelial nitric oxide synthase (eNOS) gene expression in cultured human aortic endothelial cells. RESULTS: No significant differences were observed between the vasoconstrictor responses to l-NMMA (2, 4, 8 micromol/min) before and after E(2) treatment. Comparison of E(2)-treated endothelial cells with control cells showed no significant increase in eNOS mRNA expression following either acute or chronic estradiol treatment. CONCLUSIONS: The present studies do not provide evidence for an eNOS-mediated cardioprotective response to estrogen and therefore suggest that additional mechanisms other than the endothelial NO system may have an important role in the cardiovascular effects of estrogen. PMID- 12093862 TI - TwoDay Algorithm in predicting fertile time. PMID- 12093863 TI - Different aetiological mechanisms for unexplained and endometriosis-associated infertility cannot be inferred from unstimulated IVF cycles using HCG to induce ovulation. PMID- 12093864 TI - Investigation of the infertile couple: laparoscopy after normal hysterosalpingography? PMID- 12093867 TI - The death domain kinase RIP protects thymocytes from tumor necrosis factor receptor type 2-induced cell death. AB - Fas and the tumor necrosis factor receptor (TNFR)1 regulate the programmed cell death of lymphocytes. The death domain kinase, receptor interacting protein (rip), is recruited to the TNFR1 upon receptor activation. In vitro, rip-/- fibroblasts are sensitive to TNF-induced cell death due to an impaired nuclear factor kappaB response. Because rip-/- mice die at birth, we were unable to examine the effects of a targeted rip mutation on lymphocyte survival. To address the contribution of RIP to immune homeostasis, we examined lethally irradiated mice reconstituted with rip-/- hematopoietic precursors. We observed a decrease in rip-/- thymocytes and T cells in both wild-type C57BL/6 and recombination activating gene 1-/- irradiated hosts. In contrast, the B cell and myeloid lineages are unaffected by the absence of rip. Thus, the death domain kinase rip is required for T cell development. Unlike Fas-associated death domain, rip does not regulate T cell proliferation, as rip-/- T cells respond to polyclonal activators. However, rip-deficient mice contain few viable CD4+ and CD8+ thymocytes, and rip-/- thymocytes are sensitive to TNF-induced cell death. Surprisingly, the rip-associated thymocyte apoptosis was not rescued by the absence of TNFR1, but appears to be rescued by an absence of TNFR2. Taken together, this study implicates RIP and TNFR2 in thymocyte survival. PMID- 12093866 TI - Contributions of the T cell receptor-associated CD3gamma-ITAM to thymocyte selection. AB - The immunoreceptor tyrosine-based activation motifs (ITAMs) in the CD3 chains associated with the T cell receptor (TCR) are crucial for TCR signaling. To probe the role of the CD3gamma-ITAM in T cell development, we created knock-in mice in which the CD3gamma chain of the TCR complex is replaced by a mutant signaling deficient CD3gamma chain, lacking the CD3gamma-ITAM. This mutation results in considerable impairment in positive selection in the polyclonal TCR repertoire. When CD3gamma-deltaITAM mice are crossed to mice expressing transgenic F5 TCRs, their thymocytes are completely unable to perform positive selection in vivo in response to intrathymic ligands. Also, the in vitro positive selection response of double-positive (DP) thymocytes with F5-CD3gamma-deltaITAM mutant receptors to their agonist ligand and many of its variants is severely impaired or abrogated. Yet, the binding and dissociation constants of agonist ligands for the F5 receptor are not affected by the CD3gamma-deltaITAM mutation. Furthermore, DP thymocytes with mutant receptors can respond to agonist ligand with normal antigen sensitivity and to normal levels, as shown by their ability to induce CD69 up-regulation, TCR down-regulation, negative selection, and ZAP70 and c-Jun NH2-terminal kinase activation. In sharp contrast, induction of extracellular signal-regulated kinase (ERK) activation and linker for activation of T cells (LAT) phosphorylation are severely impaired in these cells. Together, these findings underscore that intrinsic properties of the TCR-CD3 complex regulate selection at the DP checkpoint. More importantly, this analysis provides the first direct genetic evidence for a role of the CD3gamma-ITAM in TCR-driven thymocyte selection. PMID- 12093868 TI - Proteolytic processing of Stat6 signaling in mast cells as a negative regulatory mechanism. AB - Accumulating evidence has shown the importance of Stat6-mediated signaling in allergic diseases. In this study, we show a novel regulatory mechanism of Stat6 mediated signaling in mast cells. When Stat6 is activated by interleukin (IL)-4 and translocated to the nucleus, Stat6 is cleaved by a nucleus-associated protease in mast cells. The cleaved 65-kD Stat6 lacks the COOH-terminal transactivation domain and functions as a dominant-negative molecule to Stat6 mediated transcription. The retrovirus-mediated expression of cleavage-resistant Stat6 mutants prolongs the nuclear accumulation of Stat6 upon IL-4 stimulation and enhances IL-4-induced gene expression and growth inhibition in mast cells. These results indicate that the proteolytic processing of Stat6 functions as a lineage-specific negative regulator of Stat6-dependent signaling in mast cells, and thus suggest that it may account for the limited role of Stat6 in IL-4 signaling in mast cells. PMID- 12093869 TI - Induction of NFATc2 expression by interleukin 6 promotes T helper type 2 differentiation. AB - Interleukin (IL)-6 is produced by professional antigen-presenting cells (APCs) such as B cells, macrophages, and dendritic cells. It has been previously shown that APC-derived IL-6 promotes the differentiation of naive CD4+ T cells into effector T helper type 2 (Th2) cells. Here, we have studied the molecular mechanism for IL-6-mediated Th2 differentiation. During the activation of CD4+ T cells, IL-6 induces the production of IL-4, which promotes the differentiation of these cells into effector Th2 cells. Regulation of IL-4 gene expression by IL-6 is mediated by nuclear factor of activated T cells (NFAT), as inhibition of NFAT prevents IL-6-driven IL-4 production and Th2 differentiation. IL-6 upregulates NFAT transcriptional activity by increasing the levels of NFATc2. The ability of IL-6 to promote Th2 differentiation is impaired in CD4+ T cells that lack NFATc2, demonstrating that NFATc2 is required for regulation of IL-4 gene expression by IL-6. Regulation of NFATc2 expression and NFAT transcriptional activity represents a novel pathway by which IL-6 can modulate gene expression. PMID- 12093870 TI - Cbl-b positively regulates Btk-mediated activation of phospholipase C-gamma2 in B cells. AB - Genetic studies have revealed that Cbl-b plays a negative role in the antigen receptor-mediated proliferation of lymphocytes. However, we show that Cbl-b deficient DT40 B cells display reduced phospholipase C (PLC)-gamma2 activation and Ca2+ mobilization upon B cell receptor (BCR) stimulation. In addition, the overexpression of Cbl-b in WEHI-231 mouse B cells resulted in the augmentation of BCR-induced Ca2+ mobilization. Cbl-b interacted with PLC-gamma2 and helped the association of PLC-gamma2 with Bruton's tyrosine kinase (Btk), as well as B cell linker protein (BLNK). Cbl-b was indispensable for Btk-dependent sustained increase in intracellular Ca2+. Both NH(2)-terminal tyrosine kinase-binding domain and COOH-terminal half region of Cbl-b were essential for its association with PLC-gamma2 and the regulation of Ca2+ mobilization. These results demonstrate that Cbl-b positively regulates BCR-mediated Ca2+ signaling, most likely by influencing the Btk/BLNK/PLC-gamma2 complex formation. PMID- 12093871 TI - Chemokine requirements for B cell entry to lymph nodes and Peyer's patches. AB - B cell entry to lymph nodes and Peyer's patches depends on chemokine receptor signaling, but the principal chemokine involved has not been defined. Here we show that the homing of CXCR4-/- B cells is suppressed in CCL19 (ELC)- and CCL21 (SLC)-deficient paucity of lymph node T cells mice, but not in wild-type mice. We also find that CXCR4 can contribute to T cell homing. Using intravital microscopy, we find that B cell adhesion to high endothelial venules (HEVs) is disrupted when CCR7 and CXCR4 are predesensitized. In Peyer's patches, B cell entry is dependent on CXCR5 in addition to CCR7/CXCR4. CXCL12 (SDF1) is displayed broadly on HEVs, whereas CXCL13 (BLC) is found selectively on Peyer's patch follicular HEVs. These findings establish the principal chemokine and chemokine receptor requirements for B cell entry to lymph nodes and Peyer's patches. PMID- 12093873 TI - T cell costimulation through CD28 depends on induction of the Bcl-xgamma isoform: analysis of Bcl-xgamma-deficient mice. AB - The molecular basis of CD28-dependent costimulation of T cells is poorly understood. Bcl-xgamma is a member of the Bcl-x family whose expression is restricted to activated T cells and requires CD28-dependent ligation for full expression. We report that Bcl-xgamma-deficient (Bcl-xgamma-/-) T cells display defective proliferative and cytokine responses to CD28-dependent costimulatory signals, impaired memory responses to proteolipid protein peptide (PLP), and do not develop PLP-induced experimental autoimmune encephalomyelitis (EAE). In contrast, enforced expression of Bcl-xgamma largely replaces the requirement for B7-dependent ligation of CD28. These findings identify the Bcl-xgamma cytosolic protein as an essential downstream link in the CD28-dependent signaling pathway that underlies T cell costimulation. PMID- 12093872 TI - Critical roles for interleukin 1 and tumor necrosis factor alpha in antibody induced arthritis. AB - In spontaneous inflammatory arthritis of K/BxN T cell receptor transgenic mice, the effector phase of the disease is provoked by binding of immunoglobulins (Igs) to joint surfaces. Inflammatory cytokines are known to be involved in human inflammatory arthritis, in particular rheumatoid arthritis, although, overall, the pathogenetic mechanisms of the human affliction remain unclear. To explore the analogy between the K/BxN model and human patients, we assessed the role and relative importance of inflammatory cytokines in K/BxN joint inflammation by transferring arthritogenic serum into a panel of genetically deficient recipients. Interleukin (IL)-1 proved absolutely necessary. Tumor necrosis factor (TNF)-alpha was also required, although seemingly less critically than IL-1, because a proportion of TNF-alpha-deficient mice developed robust disease. There was no evidence for an important role for IL-6. Bone destruction and reconstruction were also examined. We found that all mice with strong inflammation exhibited the bone erosion and reconstruction phenomena typical of K/BxN arthritis, with no evidence of any particular requirement for TNFalpha for bone destruction. The variability in the requirement for TNF-alpha, reminiscent of that observed in treated rheumatoid arthritis patients, did not appear genetically programmed but related instead to subtle environmental changes. PMID- 12093874 TI - Constitutive expression of the B7h ligand for inducible costimulator on naive B cells is extinguished after activation by distinct B cell receptor and interleukin 4 receptor-mediated pathways and can be rescued by CD40 signaling. AB - The recently described ligand-receptor pair, B7h-inducible costimulator (ICOS), is critical for germinal center formation and antibody responses. In contrast to the induced expression of the related costimulatory ligands B7.1 and B7.2, B7h is constitutively expressed on naive B cells and is surprisingly extinguished after antigen engagement and interleukin (IL)-4 cytokine signaling. Although signaling through both B cell receptor (BCR) and IL-4 receptor (R) converge on the extinction of B7h mRNA levels, BCR down-regulation occurs through Ca2+ mobilization, whereas IL-4R down-regulation occurs through a distinct Stat6 dependent pathway. During antigen-specific B cell activation, costimulation through CD40 signaling can reverse both BCR- and IL-4R-mediated B7h down regulation. These data suggest that the CD40-CD40 ligand signaling pathway regulates B7h expression on activated B cells and may control whether antigen activated B cells can express B7h and costimulate cognate antigen-activated T cells through ICOS. PMID- 12093875 TI - Multidrug efflux systems play an important role in the invasiveness of Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa is an important opportunistic human pathogen. Certain strains can transmigrate across epithelial cells, and their invasive phenotype is correlated with capacity to cause invasive human disease and fatal septicemia in mice. Four multidrug efflux systems have been described in P. aeruginosa, however, their contribution to virulence is unclear. To clarify the role of efflux systems in invasiveness, P. aeruginosa PAO1 wild-type (WT) and its efflux mutants were evaluated in a Madin-Darby canine kidney (MDCK) epithelial cell monolayer system and in a murine model of endogenous septicemia. All efflux mutants except a deltamexCD-oprJ deletion demonstrated significantly reduced invasiveness compared with WT. In particular, a deltamexAB-oprM deletion strain was compromised in its capacity to invade or transmigrate across MDCK cells, and could not kill mice, in contrast to WT which was highly invasive (P < 0.0006) and caused fatal infection (P < 0.0001). The other mutants, including deltamexB and deltamexXY mutants, were intermediate between WT and the deltamexAB-oprM mutant in invasiveness and murine virulence. Invasiveness was restored to the deltamexAB oprM mutant by complementation with mexAB-oprM or by addition of culture supernatant from MDCK cells infected with WT. We conclude that the P. aeruginosa MexAB-OprM efflux system exports virulence determinants that contribute to bacterial virulence. PMID- 12093876 TI - A critical role for natural killer T cells in immunosurveillance of methylcholanthrene-induced sarcomas. AB - Natural killer (NK) T cells initiate potent antitumor responses when stimulated by exogenous factors such as interleukin (IL)-12 or alpha-galactosylceramide (alpha-GalCer), however, it is not clear whether this reflects a physiological role for these cells in tumor immunity. Through adoptive transfer of NK T cells from wild-type to NK T cell-deficient (T cell receptor [TCR] Jalpha281-/-) mice, we demonstrate a critical role for NK T cells in immunosurveillance of methylcholanthrene (MCA)-induced fibrosarcomas, in the absence of exogenous stimulatory factors. Using the same approach with gene-targeted and/or antibody depleted donor or recipient mice, we have shown that this effect depends on CD1d recognition and requires the additional involvement of both NK and CD8+ T cells. Interferon-gamma production by both NK T cells and downstream, non-NK T cells, is essential for protection, and perforin production by effector cells, but not NK T cells, is also critical. The protective mechanisms in this more physiologically relevant system are distinct from those associated with alpha-GalCer-induced, NK T cell-mediated, tumor rejection. This study demonstrates that, in addition to their importance in tumor immunotherapy induced by IL-12 or alpha-GalCer, NK T cells can play a critical role in tumor immunosurveillance, at least against MCA induced sarcomas, in the absence of exogenous stimulation. PMID- 12093877 TI - Suppression of lymphoma and epithelial malignancies effected by interferon gamma. AB - The immunosurveillance of transformed cells by the immune system remains one of the most controversial and poorly understood areas of immunity. Gene-targeted mice have greatly aided our understanding of the key effector molecules in tumor immunity. Herein, we describe spontaneous tumor development in gene-targeted mice lacking interferon (IFN)-gamma and/or perforin (pfp), or the immunoregulatory cytokines, interleukin (IL)-12, IL-18, and tumor necrosis factor (TNF). Both IFN gamma and pfp were critical for suppression of lymphomagenesis, however the level of protection afforded by IFN-gamma was strain specific. Lymphomas arising in IFN gamma-deficient mice were very nonimmunogenic compared with those derived from pfp-deficient mice, suggesting a comparatively weaker immunoselection pressure by IFN-gamma. Single loss of IL-12, IL-18, or TNF was not sufficient for spontaneous tumor development. A significant incidence of late onset adenocarcinoma observed in both IFN-gamma- and pfp-deficient mice indicated that some epithelial tissues were also subject to immunosurveillance. PMID- 12093878 TI - Delayed apoptotic cell clearance and lupus-like autoimmunity in mice lacking the c-mer membrane tyrosine kinase. AB - Mice lacking the membrane tyrosine kinase c-mer have been shown to have altered macro-phage cytokine production and defective phagocytosis of apoptotic cells despite normal phagocytosis of other particles. We show here that c-mer-deficient mice have impaired clearance of infused apoptotic cells and that they develop progressive lupus-like autoimmunity, with antibodies to chromatin, DNA, and IgG. The autoimmunity appears to be driven by endogenous antigens, with little polyclonal B cell activation. These mice should be an excellent model for studying the role of apoptotic debris as an immunogenic stimulus for systemic autoimmunity. PMID- 12093879 TI - Activation and differentiation of autoreactive B-1 cells by interleukin 10 induce autoimmune hemolytic anemia in Fas-deficient antierythrocyte immunoglobulin transgenic mice. AB - The Fas (CD95) gene is among critical genetic factors in some autoimmune diseases, which are characterized by autoantibody (autoAb) productions. In mice, mutations in the Fas gene cause lymphoproliferation (lpr) which predominantly develops glomerulonephritis, whereas the mutations in human cause autoimmune lymphoproliferative syndrome (ALPS) characterized by autoimmune hemolytic anemia (AIHA) and thrombocytopenia. Although the mechanism of antinuclear Ab in Fas deficient background has been well characterized, that of antierythrocyte Ab production in ALPS has been still unclear. To investigate this mechanism, we developed a mouse line by crossing the antierythrocyte antibody transgenic mice (H+L6 mice) and Fas-deficient mice. Although Fas deficiency did not break tolerance of autoreactive B-2 cells in H+L6 mice, autoreactive B-1 cells in Fas deficient H+L6 homozygous mice became activated and differentiated into autoAb producing cells in mesenteric lymph nodes and lamina propria of intestine, resulting in severe anemia. In addition, serum levels of interleukin (IL)-10 significantly increased in Fas-/- x H+L6 homozygous mice and administration of anti-IL-10 Ab prevented exacerbation of autoAb production and AIHA. These results suggest that activation of B-1 cells is responsible for induction of AIHA in Fas deficient condition and that IL-10 plays a critical role in terminal differentiation of B-1 cells in these mice. PMID- 12093880 TI - The therapeutic potential of modulating the ceramide/sphingomyelin pathway. PMID- 12093881 TI - Influenza vaccines: present and future. PMID- 12093882 TI - Strategies for an HIV vaccine. PMID- 12093883 TI - Live-attenuated virus vaccines for respiratory syncytial and parainfluenza viruses: applications of reverse genetics. PMID- 12093884 TI - Human CYP7A1 deficiency: progress and enigmas. PMID- 12093885 TI - The choreography of cyclooxygenases in the kidney. PMID- 12093887 TI - Glucagon-like peptide-1 receptor stimulation increases blood pressure and heart rate and activates autonomic regulatory neurons. AB - Glucagon-like peptide-1 (GLP-1) released from the gut functions as an incretin that stimulates insulin secretion. GLP-1 is also a brain neuropeptide that controls feeding and drinking behavior and gastric emptying and elicits neuroendocrine responses including development of conditioned taste aversion. Although GLP-1 receptor (GLP-1R) agonists are under development for the treatment of diabetes, GLP-1 administration may increase blood pressure and heart rate in vivo. We report here that centrally and peripherally administered GLP-1R agonists dose-dependently increased blood pressure and heart rate. GLP-1R activation induced c-fos expression in the adrenal medulla and neurons in autonomic control sites in the rat brain, including medullary catecholamine neurons providing input to sympathetic preganglionic neurons. Furthermore, GLP-1R agonists rapidly activated tyrosine hydroxylase transcription in brainstem catecholamine neurons. These findings suggest that the central GLP-1 system represents a regulator of sympathetic outflow leading to downstream activation of cardiovascular responses in vivo. PMID- 12093886 TI - Increased ABCA1 activity protects against atherosclerosis. AB - The ABC transporter ABCA1 plays a key role in the first steps of the reverse cholesterol transport pathway by mediating lipid efflux from macrophages. Previously, it was demonstrated that human ABCA1 overexpression in vivo in transgenic mice results in a mild elevation of plasma HDL levels and increased efflux of cholesterol from macrophages. In this study, we determined the effect of overexpression of ABCA1 on atherosclerosis development. Human ABCA1 transgenic mice (BAC(+)) were crossed with ApoE(-/-) mice, a strain that spontaneously develop atherosclerotic lesions. BAC(+)ApoE(-/-) mice developed dramatically smaller, less-complex lesions as compared with their ApoE(-/-) counterparts. In addition, there was increased efflux of cholesterol from macrophages isolated from the BAC(+)ApoE(-/-) mice. Although the increase in plasma HDL cholesterol levels was small, HDL particles from BAC(+)ApoE(-/-) mice were significantly better acceptors of cholesterol. Lipid analysis of HDL particles from BAC(+)ApoE( /-) mice revealed an increase in phospholipid levels, which was correlated significantly with their ability to enhance cholesterol efflux. PMID- 12093888 TI - Molecular mechanisms of blister formation in bullous impetigo and staphylococcal scalded skin syndrome. AB - Bullous impetigo due to Staphylococcus aureus is one of the most common bacterial infections of man, and its generalized form, staphylococcal scalded skin syndrome (SSSS), is a frequent manifestation of staphylococcal epidemics in neonatal nurseries. Both diseases are mediated by exfoliative toxins (ETs), which show exquisite pathologic specificity in blistering only the superficial epidermis. We show that these toxins act as serine proteases with extremely focused molecular specificity to cleave mouse and human desmoglein 1 (Dsg1) once after glutamic acid residue 381 between extracellular domains 3 and 4. Mutation of the predicted catalytically active serine to alanine completely inhibits cleavage. The mutated ETs bind specifically to Dsg1 by immunofluorescence colocalization and by coimmunoprecipitation. Thus, ETs, through specific recognition and proteolytic cleavage of one structurally critical peptide bond in an adhesion molecule, cause its dysfunction and allow S. aureus to spread under the stratum corneum, the main barrier of the skin, explaining how, although they circulate through the entire body in SSSS, they cause pathology only in the superficial epidermis. PMID- 12093889 TI - Opposite effects of cyclooxygenase-1 and -2 activity on the pressor response to angiotensin II. AB - Therapeutic use of cyclooxygenase-inhibiting (COX-inhibiting) nonsteroidal antiinflammatory drugs (NSAIDs) is often complicated by renal side effects including hypertension and edema. The present studies were undertaken to elucidate the roles of COX1 and COX2 in regulating blood pressure and renal function. COX2 inhibitors or gene knockout dramatically augment the pressor effect of angiotensin II (Ang II). Unexpectedly, after a brief increase, the pressor effect of Ang II was abolished by COX1 deficiency (either inhibitor or knockout). Ang II infusion also reduced medullary blood flow in COX2-deficient but not in control or COX1-deficient animals, suggesting synthesis of COX2 dependent vasodilators in the renal medulla. Consistent with this, Ang II failed to stimulate renal medullary prostaglandin E(2) and prostaglandin I(2) production in COX2-deficient animals. Ang II infusion normally promotes natriuresis and diuresis, but COX2 deficiency blocked this effect. Thus, COX1 and COX2 exert opposite effects on systemic blood pressure and renal function. COX2 inhibitors reduce renal medullary blood flow, decrease urine flow, and enhance the pressor effect of Ang II. In contrast, the pressor effect of Ang II is blunted by COX1 inhibition. These results suggest that, rather than having similar cardiovascular effects, the activities of COX1 and COX2 are functionally antagonistic. PMID- 12093891 TI - Multiantigen/multiepitope-directed immune-specific suppression of "complex autoimmune encephalomyelitis" by a novel protein product of a synthetic gene. AB - Systemic administration of antigen/peptide for peripheral T cell tolerance has long been investigated as a potential approach to therapy of autoimmune diseases. The multiple antimyelin T cell reactivities likely to be associated with multiple sclerosis (MS) impose major difficulties in devising such an immune-specific therapeutic approach to the disease, because targeting T cells specific for a single autoantigen/epitope is unlikely to be sufficiently effective. Here, we present a pilot study on the possibility of concomitantly inhibiting multiple potentially pathogenic antimyelin T cell reactivities by tolerogenic administration of an artificial "multiantigen/multiepitope" protein. A synthetic gene was constructed to encode selected disease-relevant epitopes of myelin basic protein (MBP), proteolipid protein (PLP), and myelin oligodendrocyte glycoprotein (MOG). The protein product, hmTAP (synthetic human multitarget autoantigen protein), was adequately processed for antigenic presentation of the relevant integral epitopes, in vitro and in vivo. Systemic administration of hmTAP not only suppressed and treated experimental autoimmune encephalomyelitis (EAE) initiated by autoreactivity to a PLP epitope, but also abrogated complex EAE transferred by multispecific line T cells reactive against encephalitogenic epitopes of MBP, PLP, and MOG. These data indicate that multiantigen/multiepitope directed therapy of complex autoimmune diseases is effective and can be mediated by the protein product of a specifically designed synthetic gene. PMID- 12093890 TI - Immune complex-mediated antigen presentation induces tumor immunity. AB - Antigen uptake receptors on dendritic cells (DCs) provide efficient entry for the initiation of antigen-specific adaptive immunity. Here we show that targeting of antigen to Fc receptors on DCs accomplishes combined activation of Th1 CD4 and CD8 effector responses in vivo, namely delayed-type hypersensitivity and tumor immunity. Tumor immunity specific for ovalbumin-expressing tumors was provided by immunization with wild-type but not FcgammaRgamma(-/-) DCs loaded with ovalbumin containing immune complexes. Tumor protection was eliminated when immune complex loaded DCs lacked beta(2) microglobulin, TAP, or MHC class II, demonstrating that Fc receptor-targeted antigenic uptake led to both MHC class I- and class II restricted responses, which together are required for effector tumor immunity. Thus the cross-presentation pathway accessed by antigens acquired endocytically through Fc receptors links humoral and cellular immunity. These data suggest that administration of antitumor antibodies may enhance tumor-specific T cell responses in vivo and provide the rationale for Fc receptor targeting in vaccine development. PMID- 12093892 TI - G(1) and G(2) cell-cycle arrest following microtubule depolymerization in human breast cancer cells. AB - Microtubule-depolymerizing agents are widely used to synchronize cells, screen for mitotic checkpoint defects, and treat cancer. The present study evaluated the effects of these agents on normal and malignant human breast cell lines. After treatment with 1 microM nocodazole, seven of ten breast cancer lines (type A cells) arrested in mitosis, whereas the other three (type B cells) did not. Similar effects were observed with 100 nM vincristine or colchicine. Among five normal mammary epithelial isolates, four exhibited type A behavior and one exhibited type B behavior. Further experiments revealed that the type B cells exhibited a biphasic dose-response curve, with mitotic arrest at low drug concentrations (100 nM nocodazole or 6 nM vincristine) that failed to depolymerize microtubules and a p53-independent p21(waf1/cip1)-associated G(1) and G(2) arrest at higher concentrations (1 microM nocodazole or 100 nM vincristine) that depolymerized microtubules. Collectively, these observations provide evidence for coupling of premitotic cell-cycle progression to microtubule integrity in some breast cancer cell lines (representing a possible "microtubule integrity checkpoint") and suggest a potential explanation for the recently reported failure of some cancer cell lines to undergo nocodazole-induced mitotic arrest despite intact mitotic checkpoint proteins. PMID- 12093893 TI - Increased C5a receptor expression in sepsis. AB - Excessive production of the complement activation product C5a appears to be harmful during the development of sepsis in rodents. Little is known about the role of the C5a receptor (C5aR) and its presence in different organs during sepsis. Using the cecal ligation/puncture (CLP) model in mice, we show here that C5aR immunoreactivity was strikingly increased in lung, liver, kidney, and heart early in sepsis in both control and neutrophil-depleted mice. C5aR mRNA expression in these organs was also significantly increased during sepsis. Immunohistochemical analysis revealed patterns of increased C5aR expression in parenchymal cells in all four organs following CLP. Mice injected at the start of CLP with a blocking IgG to C5aR (alphaC5aR) showed dramatically improved survival when compared with animals receiving nonspecific IgG, as did mice injected with alphaC5a. In alphaC5aR-treated mice, serum levels of IL-6 and TNF-alpha and bacterial counts in various organs were significantly reduced during CLP when compared with control CLP animals. These studies demonstrate for the first time that C5aR is upregulated in lung, liver, kidney, and heart during the early phases of sepsis and that blockade of C5aR is highly protective from the lethal outcome of sepsis. PMID- 12093894 TI - Human cholesterol 7alpha-hydroxylase (CYP7A1) deficiency has a hypercholesterolemic phenotype. AB - Bile acid synthesis plays a critical role in the maintenance of mammalian cholesterol homeostasis. The CYP7A1 gene encodes the enzyme cholesterol 7alpha hydroxylase, which catalyzes the initial step in cholesterol catabolism and bile acid synthesis. We report here a new metabolic disorder presenting with hyperlipidemia caused by a homozygous deletion mutation in CYP7A1. The mutation leads to a frameshift (L413fsX414) that results in loss of the active site and enzyme function. High levels of LDL cholesterol were seen in three homozygous subjects. Analysis of a liver biopsy and stool from one of these subjects revealed double the normal hepatic cholesterol content, a markedly deficient rate of bile acid excretion, and evidence for upregulation of the alternative bile acid pathway. Two male subjects studied had hypertriglyceridemia and premature gallstone disease, and their LDL cholesterol levels were noticeably resistant to 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors. One subject also had premature coronary and peripheral vascular disease. Study of the kindred, which is of English and Celtic background, revealed that individuals heterozygous for the mutation are also hyperlipidemic, indicating that this is a codominant disorder. PMID- 12093895 TI - A translational rheostat for RFLAT-1 regulates RANTES expression in T lymphocytes. AB - Activation of T lymphocytes by specific antigen triggers a 3- to 7-day maturation process. Terminal differentiation begins late after T cell activation and involves expression of effector genes, including the chemokine RANTES and its major transcriptional regulator, RANTES factor of late-activated T lymphocytes-1 (RFLAT-1). In this article we demonstrate that RFLAT-1 expression is translationally regulated through its 5'-UTR and in a cell type-specific manner. Overexpression of the translation initiation factor eIF4E increases RFLAT-1 protein, while inhibition of Mnk1, which phosphorylates eIF4E, reduces RFLAT-1 production, indicating cap-dependent translational regulation. These events are regulated by ERK-1/2 and p38 MAP kinases and allow T cells to rapidly adjust RANTES expression in response to changes in the cellular environment, such as stress and/or growth factors. These findings provide a molecular mechanism for a rheostat effect of increasing or decreasing RANTES expression at sites of inflammation. Memory T cells, already poised to make RANTES, are finely regulated by translational control of the major transcription factor regulating RANTES expression. This is the first example of such a mechanism regulating a chemokine, but it seems likely that this will prove to be a general way for cells to rapidly respond to stress, cytokines, and other proinflammatory factors in their local environment. PMID- 12093896 TI - Heparin's anti-inflammatory effects require glucosamine 6-O-sulfation and are mediated by blockade of L- and P-selectins. AB - Heparin has been used clinically as an anticoagulant and antithrombotic agent for over 60 years. Here we show that the potent anti-inflammatory property of heparin results primarily from blockade of P-selectin and L-selectin. Unfractionated heparin and chemically modified analogs were tested as inhibitors of selectin binding to immobilized sialyl Lewis(X) and of cell adhesion to immobilized selectins or thrombin-activated endothelial cells. Compared with unfractionated heparin, the modified heparinoids had inhibitory activity in this general order: over-O-sulfated heparin > heparin > 2-O,3-O-desulfated > or = N-desulfated/N acetylated heparin > or = carboxyl-reduced heparin > or= N-,2-O,3-O-desulfated heparin >> 6-O-desulfated heparin. The heparinoids also showed similar differences in their ability to inhibit thioglycollate-induced peritonitis and oxazolone-induced delayed-type hypersensitivity. Mice deficient in P- or L selectins showed impaired inflammation, which could be further reduced by heparin. However, heparin had no additional effect in mice deficient in both P- and L-selectins. We conclude that (a) heparin's anti-inflammatory effects are mainly mediated by blocking P- and L-selectin-initiated cell adhesion; (b) the sulfate groups at C6 on the glucosamine residues play a critical role in selectin inhibition; and (c) some non-anticoagulant forms of heparin retain anti inflammatory activity. Such analogs may prove useful as therapeutically effective inhibitors of inflammation. PMID- 12093897 TI - Selective interaction of plant homeodomain proteins mediates high DNA-binding affinity. AB - Understanding molecular mechanisms that control cell fate in the shoot apical meristem is a fundamental question in plant development. Genetic and molecular studies demonstrate that maize KNOTTED1 (KN1) of the TALE (3-aa acid loop extension) class of homeodomain (HD) proteins is involved in shoot apical meristem function. We show that KN1 interacts with knotted interacting protein (KIP), a BEL1-like TALE HD protein. Interaction between KN1 and KIP is mediated by conserved domains in the N termini of both proteins. The KN1 DNA-binding sequence, TGACAG(G/C)T, was biochemically identified, and in vitro DNA-binding assays show that individually KN1 and the HD of KIP bind specifically to this motif with low affinity. The KN1-KIP complex, however, binds specifically to this DNA-binding motif with high affinity, indicating that the association of KN1 and KIP may function in transcriptional regulation. PMID- 12093898 TI - (1)H and (13)C MAS NMR evidence for pronounced ligand-protein interactions involving the ionone ring of the retinylidene chromophore in rhodopsin. AB - Rhodopsin is a member of the superfamily of G-protein-coupled receptors. This seven alpha-helix transmembrane protein is the visual pigment of the vertebrate rod photoreceptor cells that mediate dim light vision. In the active binding site of this protein the ligand or chromophore, 11-cis-retinal, is covalently bound via a protonated Schiff base to lysine residue 296. Here we present the complete (1)H and (13)C assignments of the 11-cis-retinylidene chromophore in its ligand binding site determined with ultra high field magic angle spinning NMR. Native bovine opsin was regenerated with 99% enriched uniformly (13)C-labeled 11-cis retinal. From the labeled pigment, (13)C carbon chemical shifts could be obtained by using two-dimensional radio frequency-driven dipolar recoupling in a solid state magic angle spinning homonuclear correlation experiment. The (1)H chemical shifts were assigned by two-dimensional heteronuclear ((1)H-(13)C) dipolar correlation spectroscopy with phase-modulated Lee-Goldburg homonuclear (1)H decoupling applied during the t(1) period. The data indicate nonbonding interactions between the protons of the methyl groups of the retinylidene ionone ring and the protein. These nonbonding interactions are attributed to nearby aromatic acid residues Phe-208, Phe-212, and Trp-265 that are in close contact with, respectively, H-16/H-17 and H-18. Furthermore, binding of the chromophore involves a chiral selection of the ring conformation, resulting in equatorial and axial positions for CH(3)-16 and CH(3)-17. PMID- 12093899 TI - Mutations in APC, Kirsten-ras, and p53--alternative genetic pathways to colorectal cancer. AB - Colorectal cancer is one of the most significant causes of cancer death. A genetic model for colorectal cancer has been proposed in which the sequential accumulation of mutations in specific genes, including adenomatous polyposis coli (APC), Kirsten-ras (K-ras), and p53, drives the transition from healthy colonic epithelia through increasingly dysplastic adenoma to colorectal cancer. We have characterized tumor mutation spectra in a large cohort of colorectal cancer patients. In marked contrast to the predictions of the sequential model of mutation accumulation, only 6.6% of tumors were found to contain mutations in APC, K-ras, and p53, with 38.7% of tumors containing mutations in only one of these genes. The most common combination of mutations was p53 and APC (27.1%), whereas mutations in both p53 and K-ras were extremely rare. Statistical analysis (two-sided Fisher's exact test) confirmed that mutations in K-ras and p53 co occurred less frequently than expected by chance (P < 0.01, Fisher's exact test). This finding suggests that these mutations lie on alternate pathways of colorectal tumor development. The heterogeneous pattern of tumor mutations in our patient cohort suggests that multiple alternative genetic pathways to colorectal cancer exist and that the widely accepted genetic model of cancer development is not representative of the majority of colorectal tumors. PMID- 12093900 TI - High-resolution structures of a chitinase complexed with natural product cyclopentapeptide inhibitors: mimicry of carbohydrate substrate. AB - Over the past years, family 18 chitinases have been validated as potential targets for the design of drugs against human pathogens that contain or interact with chitin during their normal life cycles. Thus far, only one potent chitinase inhibitor has been described in detail, the pseudotrisaccharide allosamidin. Recently, however, two potent natural-product cyclopentapeptide chitinase inhibitors, argifin and argadin, were reported. Here, we describe high-resolution crystal structures that reveal the details of the interactions of these cyclopeptides with a family 18 chitinase. The structures are examples of complexes of a carbohydrate-processing enzyme with high-affinity peptide-based inhibitors and show in detail how the peptide backbone and side chains mimic the interactions of the enzyme with chitooligosaccharides. Together with enzymological characterization, the structures explain why argadin shows an order of magnitude stronger inhibition than allosamidin, whereas argifin shows weaker inhibition. The peptides bind to the chitinase in remarkably different ways, which may explain the differences in inhibition constants. The two complexes provide a basis for structure-based design of potent chitinase inhibitors, accessible by standard peptide chemistry. PMID- 12093903 TI - An approach for global scanning of single nucleotide variations. AB - Efficient global scanning of single nucleotide variations in DNA sequences between related, complex DNA samples remains a challenge. In the present article we present an approach to this problem. We have used immobilized thymidine DNA glycosylases to capture and enrich DNA fragments containing internal mismatched base pairs and separate these fragments as a pool from perfectly base-paired fragments as another pool. Enrichments of up to several hundredfold were obtained with one cycle of treatment, and all of the four groups of single nucleotide mismatches were fully covered by combining use of two thymine DNA glycosylases generated here. We have used a heterohybrid-orientating strategy for selective amplification of duplexes with one strand derived from each of two input DNA samples, which can also be used for selective amplification of duplexes with both strands derived from one of two input samples when desired. By combining these methods, the single nucleotide variations either between two DNA pools or within one DNA pool can be obtained in one process. This approach has been applied to the total cDNA from a human cell line and has several potential applications in mapping genetic variations, particularly global scanning of cDNA single nucleotide variations or polymorphisms, and finally high-throughput mapping of complex genetic traits. PMID- 12093902 TI - The crystal structure of a tetrameric hemoglobin in a partial hemichrome state. AB - Tetrameric hemoglobins are the most widely used systems in studying protein cooperativity. Allosteric effects in hemoglobins arise from the switch between a relaxed (R) state and a tense (T) state occurring upon oxygen release. Here we report the 2.0-A crystal structure of the main hemoglobin component of the Antarctic fish Trematomus newnesi, in a partial hemichrome form. The two alpha subunit iron atoms are bound to a CO molecule, whereas in the beta subunits the distal histidine residue is the sixth ligand of the heme iron. This structure, a tetrameric hemoglobin in the hemichrome state, demonstrates that the iron coordination by the distal histidine, usually associated with denaturing states, may be tolerated in a native-like hemoglobin structure. In addition, several features of the tertiary and quaternary organization of this structure are intermediate between the R and T states and agree well with the R --> T transition state properties obtained by spectroscopic and kinetic techniques. The analysis of this structure provides a detailed pathway of heme-heme communication and it indicates that the plasticity of the beta heme pocket plays a role in the R --> T transition of tetrameric hemoglobins. PMID- 12093904 TI - Direct visualization of distinct T cell epitopes derived from a melanoma tumor associated antigen by using human recombinant antibodies with MHC- restricted T cell receptor-like specificity. AB - Specificity in the cellular immune system is controlled and regulated by the T cell antigen receptor (TCR), which specifically recognizes peptide/major histocompatibility complex (MHC) molecules. In recent years many cancer associated MHC-restricted peptides have been isolated and because of their highly restricted fine specificity, they are desirable targets for novel approaches in immunotherapy. Antibodies that would recognize tumor-associated MHC-peptide complexes with the same specificity as the TCR would be valuable reagents for studying antigen presentation by tumor cells, for visualizing MHC-peptide complexes on cells, and eventually for monitoring the expression of specific complexes during immunotherapy. To generate molecules with such a unique fine specificity, we selected a large nonimmune repertoire of phage Fab antibodies on recombinant HLA-A2 complexed with three common antigenic T cell, HLA-A2 restricted epitopes derived from the melanoma differentiation antigen gp100. We were able to isolate a surprisingly large panel of human recombinant Fab antibodies that exhibit a characteristic TCR-like binding specificity to each of the three gp100-derived epitopes, yet unlike TCRs, they did so with an affinity in the nanomolar range. These TCR-like antibodies recognize the native MHC peptide complex expressed on the surface of antigen-presenting cells. Moreover, they can detect the specific MHC-peptide complexes on the surface of melanoma tumor cells. These results demonstrate the ability to isolate high-affinity human recombinant antibodies with the antigen-specific, MHC-restricted specificity of T cells, and this ability was demonstrated for three different epitopes of the same melanoma-derived antigen. PMID- 12093901 TI - The complete genome sequence of Chlorobium tepidum TLS, a photosynthetic, anaerobic, green-sulfur bacterium. AB - The complete genome of the green-sulfur eubacterium Chlorobium tepidum TLS was determined to be a single circular chromosome of 2,154,946 bp. This represents the first genome sequence from the phylum Chlorobia, whose members perform anoxygenic photosynthesis by the reductive tricarboxylic acid cycle. Genome comparisons have identified genes in C. tepidum that are highly conserved among photosynthetic species. Many of these have no assigned function and may play novel roles in photosynthesis or photobiology. Phylogenomic analysis reveals likely duplications of genes involved in biosynthetic pathways for photosynthesis and the metabolism of sulfur and nitrogen as well as strong similarities between metabolic processes in C. tepidum and many Archaeal species. PMID- 12093905 TI - Knockout of the alpha 1A/C-adrenergic receptor subtype: the alpha 1A/C is expressed in resistance arteries and is required to maintain arterial blood pressure. AB - alpha 1-adrenergic receptors (ARs) play a major role in blood pressure regulation. The three alpha 1-AR subtypes (A/C, B, and D) stimulate contraction of isolated arteries, but it is uncertain how different subtypes contribute to blood pressure regulation in the intact animal. We studied the role of the alpha 1A/C subtype by using gene knockout. alpha 1A/C knockout (KO) mice were viable and overtly normal. The LacZ reporter gene replaced alpha 1A/C coding sequence in the KO, and beta-galactosidase staining was present in resistance arteries and arterioles, but not in the thoracic aorta or its main branches. By tail cuff manometer and arterial catheter in conscious mice, alpha 1A/C KO mice were hypotensive at rest, with an 8-12% reduction of blood pressure dependent on alpha 1A/C gene copy number. A61603, an alpha 1A/C-selective agonist, caused a pressor response that was lost in the KO and reduced but significant in heterozygous mice with a single copy of the alpha 1A/C. A subtype-nonselective agonist [phenylephrine (PE)] caused a pressor response in KO mice, but the final arterial pressure was only 85% of wild type. The baroreflex was reset in the KO, and heart rate variability was decreased. After baroreflex blockade with atropine, PE increased blood pressure but did not change heart rate. Cardiac and vascular responses to the beta-AR agonist isoproterenol were unchanged, and the arterial lumen area was not altered. We conclude that the alpha 1A/C-AR subtype is a vasopressor expressed in resistance arteries and is required for normal arterial blood pressure regulation. alpha 1A/C-selective antagonists might be desirable antihypertensive agents. PMID- 12093906 TI - Potent stimulation of transcription-coupled DNA supercoiling by sequence-specific DNA-binding proteins. AB - Transcription by RNA polymerase can stimulate localized DNA supercoiling in Escherichia coli. In vivo, there is extensive experimental support for a "twin domain" model in which positive DNA supercoils are generated ahead of a translocating RNA polymerase complex and negative supercoils are formed behind it. Negative supercoils accumulate in the template DNA because the positive supercoils are preferentially removed by cellular topoisomerase action. Yet, in vitro, clear and convincing support for the twin-domain mechanism has been lacking. In this article, we reconcile this inconsistency by showing that, in a defined in vitro system with plasmid DNA templates, a variety of sequence specific DNA-binding proteins, such as the bacteriophage lambda O replication initiator or the E. coli lactose or galactose repressors, strikingly stimulate transcription-coupled DNA supercoiling. We demonstrate further that this stimulation requires the presence in the DNA template of a recognition sequence for the relevant DNA-binding protein and depends on the production of long RNA chains by an RNA polymerase. Our data are most consistent with a model in which specific DNA-binding proteins facilitate a twin-domain mechanism to enhance DNA supercoiling during transcription. More precisely, we suggest that some nucleoprotein complexes, perhaps those that contain sharply bent DNA, can form barriers that impede the diffusion and merger of independent chromosomal supercoil domains. Localization of DNA supercoils by nucleoprotein complexes may serve as a general mechanism for modulating DNA transactions that are sensitive to DNA superhelicity. PMID- 12093907 TI - Gene order evolution and paleopolyploidy in hemiascomycete yeasts. AB - The wealth of comparative genomics data from yeast species allows the molecular evolution of these eukaryotes to be studied in great detail. We used "proximity plots" to visually compare chromosomal gene order information from 14 hemiascomycetes, including the recent Genolevures survey, to Saccharomyces cerevisiae. Contrary to the original reports, we find that the Genolevures data strongly support the hypothesis that S. cerevisiae is a degenerate polyploid. Using gene order information alone, 70% of the S. cerevisiae genome can be mapped into "sister" regions that tile together with almost no overlap. This map confirms and extends the map of sister regions that we constructed previously by using duplicated genes, an independent source of information. Combining gene order and gene duplication data assigns essentially the whole genome into sister regions, the largest gap being only 36 genes long. The 16 centromere regions of S. cerevisiae form eight pairs, indicating that an ancestor with eight chromosomes underwent complete doubling; alternatives such as segmental duplications can be ruled out. Gene arrangements in Kluyveromyces lactis and four other species agree quantitatively with what would be expected if they diverged from S. cerevisiae before its polyploidization. In contrast, Saccharomyces exiguus, Saccharomyces servazzii, and Candida glabrata show higher levels of gene adjacency conservation, and more cases of imperfect conservation, suggesting that they split from the S. cerevisiae lineage after polyploidization. This finding is confirmed by sequences around the C. glabrata TRP1 and IPP1 loci, which show that it contains sister regions derived from the same duplication event as that of S. cerevisiae. PMID- 12093908 TI - Mutations in the RNase H domain of HIV-1 reverse transcriptase affect the initiation of DNA synthesis and the specificity of RNase H cleavage in vivo. AB - Retroviral reverse transcriptases contain a DNA polymerase activity that can copy an RNA or DNA template and an RNase H activity that degrades the viral RNA genome during reverse transcription. RNase H makes both specific and nonspecific cleavages; specific cleavages are used to generate and remove the polypurine tract primer used for plus-strand DNA synthesis and to remove the tRNA primer used for minus-strand DNA synthesis. We generated mutations in an HIV-1-based vector to change amino acids in the RNase H domain that contact either the RNA and DNA strands. Some of these mutations affected the initiation of DNA synthesis, demonstrating an interdependence of the polymerase and RNase H activities of HIV-1 reverse transcription during viral DNA synthesis. The ends of the linear DNA form of the HIV-1 genome are defined by the specific RNase H cleavages that remove the plus- and minus-strand primers; these ends can be joined to form two-long-terminal repeat circles. Analysis of two-long-terminal repeat circle junctions showed that mutations in the RNase H domain affect the specificity of RNase H cleavage. PMID- 12093909 TI - Assessing the role of Ih channels in synaptic transmission and mossy fiber LTP. AB - Hyperpolarization-activated nonselective cation channels (Ih channels) play an important role in the control of membrane excitability and rhythmic neuronal activity. The functional relevance of presynaptic Ih channels in regulating synaptic function, however, is not well established. Recently, it has been proposed [Mellor, J., Nicoll, R. A. & Schmitz, D. (2002) Science 295, 143-147] that presynaptic Ih channels are necessary for hippocampal mossy fiber long-term potentiation (LTP). This observation challenges an alternative model that suggests presynaptic forms of LTP are caused by a direct modification of the transmitter release machinery. Here, we assess the role of Ih in hippocampal mossy fiber LTP as well as cerebellar parallel fiber LTP, forms of potentiation that share common mechanisms. Our results show that after Ih blockade neither mossy fiber LTP nor parallel fiber LTP are affected. Furthermore, Ih does not significantly modify basal excitatory synaptic transmission in the hippocampus, whereas the organic Ih blockers ZD7288 and DK-AH 269 induce a large Ih independent depression of synaptic transmission. In summary, our results indicate that Ih-mediated persistent changes in presynaptic excitability do not underlie presynaptic forms of LTP. PMID- 12093910 TI - Beyond the Rayleigh instability limit for multicharged finite systems: from fission to Coulomb explosion. AB - We address the stability of multicharged finite systems driven by Coulomb forces beyond the Rayleigh instability limit. Our exploration of the nuclear dynamics of heavily charged Morse clusters enabled us to vary the range of the pair potential and of the fissibility parameter, which results in distinct fragmentation patterns and in the angular distributions of the fragments. The Rayleigh instability limit separates between nearly binary (or tertiary) spatially unisotropic fission and spatially isotropic Coulomb explosion into a large number of small, ionic fragments. Implications are addressed for a broad spectrum of dynamics in chemical physics, radiation physics of ultracold gases, and biophysics, involving the fission of clusters and droplets, the realization of Coulomb explosion of molecular clusters, the isotropic expansion of optical molasses, and the Coulomb instability of "isolated" proteins. PMID- 12093911 TI - The fifth essential DNA polymerase phi in Saccharomyces cerevisiae is localized to the nucleolus and plays an important role in synthesis of rRNA. AB - We report that POL5 encodes the fifth essential DNA polymerase in Saccharomyces cerevisiae. Pol5p was identified and purified from yeast cell extracts and is an aphidicolin-sensitive DNA polymerase that is stimulated by yeast proliferating cell nuclear antigen (PCNA). Thus, we named Pol5p DNA polymerase phi. Temperature sensitive pol5-1-- -3 mutants did not arrest at G(2)/M at the restrictive temperature. Furthermore, the polymerase active-site mutant POL5dn gene complements the lethality of Delta pol5. These results suggest that the polymerase activity of Pol5p is not required for the in vivo function of Pol5p. rRNA synthesis was severely inhibited at the restrictive temperature in the temperature-sensitive pol5-3 mutant cells, suggesting that an essential function of Pol5p is rRNA synthesis. Pol5p is localized exclusively to the nucleolus and binds near or at the enhancer region of rRNA-encoding DNA repeating units. PMID- 12093912 TI - A structural basis for the activity of retro-Diels-Alder catalytic antibodies: evidence for a catalytic aromatic residue. AB - The nitroxyl synthase catalytic antibodies 10F11, 9D9, and 27C5 catalyze the release of nitroxyl from a bicyclic pro-drug by accelerating a retro-Diels-Alder reaction. The Fabs (antigen-binding fragments) of these three catalytic antibodies were cloned and sequenced. Fab 9D9 was crystallized in the apo-form and in complex with one transition state analogue of the reaction. Crystal structures of Fab 10F11 in complex with ligands mimicking substrate, transition state, and product have been determined at resolutions ranging from 1.8 to 2.3 A. Antibodies 9D9 and 10F11 show increased shape complementarity (as quantified by the program sc) to the hapten and to a modeled transition state as compared with substrate and product. The shape complementarity is mediated to a large extent by an aromatic residue (tyrosine or tryptophan) at the bottom of the hydrophobic active pocket, which undergoes pi-stacking interactions with the aromatic rings of the ligands. Another factor contributing to the different reactivity of the regioisomers probably arises because of hydrogen-bonding interactions between the nitroxyl bridge and the backbone amide of PheH101 and possibly a conserved water molecule. PMID- 12093913 TI - Promoter-proximal tethering elements regulate enhancer-promoter specificity in the Drosophila Antennapedia complex. AB - Insulator DNAs and promoter competition regulate enhancer-promoter interactions within complex genetic loci. Here we provide evidence for a third mechanism: promoter-proximal tethering elements. The Scr-ftz region of the Antennapedia gene complex includes two known enhancers, AE1 and T1. AE1 selectively interacts with the ftz promoter to maintain pair-rule stripes of ftz expression during gastrulation and germ-band elongation. The T1 enhancer, located 3' of the ftz gene and approximately 25 kb 5' of the Scr promoter, selectively activates Scr expression in the prothorax and posterior head segments. A variety of P element minigenes were examined in transgenic embryos to determine the basis for specific AE1-ftz and T1-Scr interactions. A 450-bp DNA fragment located approximately 100 bp 5' of the Scr transcription start site is essential for T1-Scr interactions and can mediate long-range activation of a ftz/lacZ reporter gene when placed 5' of the ftz promoter. We suggest that the Scr450 fragment contains tethering elements that selectively recruit T1 to the Scr promoter. Tethering elements might regulate enhancer-promoter interactions at other complex genetic loci. PMID- 12093914 TI - Erk5 null mice display multiple extraembryonic vascular and embryonic cardiovascular defects. AB - Erk5 is a mitogen-activated protein kinase, the biological role of which is largely undefined. Therefore, we deleted the erk5 gene in mice to assess its function in vivo. Inactivation of the erk5 gene resulted in defective blood vessel and cardiac development leading to embryonic lethality around embryonic days 9.5-10.5. Cardiac development was retarded largely, and the heart failed to undergo normal looping. Endothelial cells that line the developing myocardium of erk5-/- embryos displayed a disorganized, rounded morphology. Vasculogenesis occurred, but extraembryonic and embryonic blood vessels were disorganized and failed to mature. Furthermore, the investment of embryonic blood vessels with smooth muscle cells was attenuated. Together, these data define an essential role for Erk5 in cardiovascular development. Moreover, the inability of Erk5-deficient mice to form a complex vasculature suggests that Erk5 may play an important role in controlling angiogenesis. PMID- 12093915 TI - Cytotoxic T lymphocytes directed against a tumor-specific mutated antigen display similar HLA tetramer binding but distinct functional avidity and tissue distribution. AB - We have previously identified an antigen (Ag) recognized on a human large cell carcinoma of the lung by a tumor-specific cytotoxic T lymphocyte clone derived from autologous tumor infiltrating lymphocytes (TILs). The antigenic peptide is presented by HLA-A2 molecules and is encoded by a mutated alpha-actinin-4 (ACTN4) gene. In the present report, we have isolated two anti-alpha-actinin-4 T cell clones from the same patient TIL and from his peripheral blood lymphocytes (PBLs) by using tetramers of soluble HLA-A2 molecules loaded with the mutated peptide. Although all of the clones displayed similar tetramer labeling, those isolated from PBL showed lower avidity of Ag recognition and killed the specific target much less efficiently, indicating that tetramer staining does not correlate with clone avidity/tumor reactivity. T cell receptor (TCR) analysis revealed that alpha-actinin-4-reactive clones used distinct alpha and beta chain rearrangements, demonstrating TCR repertoire diversity. Interestingly, TCR beta chain gene usage indicated that only Ag-specific clones with high functional avidity were expanded at the tumor site, whereas a low-avidity clone was exclusively amplified in patient peripheral blood. Our results point to the existence of distinct but overlapping antitumor TCR repertoires in TIL and PBL and suggest a selective in situ expansion of tumor-specific cytotoxic T lymphocyte with high avidity/tumor reactivity. PMID- 12093916 TI - Evidence for a cytopathogenicity determinant in HIV-1 Vpr. AB - HIV-1 is cytopathic for CD4(+) T lymphocytes in vitro and this property of HIV-1 is generally considered to account for some of its in vivo cytopathogenicity. Thus, the extent of lymphocyte depletion correlates with the level of viremia whereas low levels of viral replication are typically associated with stable lymphocyte levels and asymptomatic infection such as is observed in non progressors. Here, we describe a non-progressor who did not fit this general pattern in that CD4(+) T lymphocyte homeostasis was maintained in the face of high-level viral replication. Biological viral isolates from this patient replicated in primary lymphocytes without inducing cytopathicity. Because this phenotype is reminiscent of Vpr-deleted viruses, we examined the contribution of the Vpr gene to the viral phenotype. Vpr alleles derived from this patient contained both premature stop codons and an unusual Q3R polymorphism. Insertion of patient-derived Vpr alleles or a Q3R substitution into a cytopathic HIV-1 clone resulted in a marked impairment of cytopathicity without affecting viral replication efficiency. The effect of Vpr on cytopathicity was unrelated to reported activities of Vpr including virion association, interaction with uracil DNA glycosylase, G(2) arrest, or enhancement of macrophage infection but correlated with the ability of Vpr to induce host cell apoptosis. This study suggests the presence of a determinant of in vivo cytopathogenicity within HIV-1 Vpr and further indicates that viral replication can be uncoupled from cytopathicity in vitro and in vivo. PMID- 12093917 TI - The protofilament structure of insulin amyloid fibrils. AB - Under solution conditions where the native state is destabilized, the largely helical polypeptide hormone insulin readily aggregates to form amyloid fibrils with a characteristic cross-beta structure. However, there is a lack of information relating the 4.8 A beta-strand repeat to the higher order assembly of amyloid fibrils. We have used cryo-electron microscopy (EM), combining single particle analysis and helical reconstruction, to characterize these fibrils and to study the three-dimensional (3D) arrangement of their component protofilaments. Low-resolution 3D structures of fibrils containing 2, 4, and 6 protofilaments reveal a characteristic, compact shape of the insulin protofilament. Considerations of protofilament packing indicate that the cross beta ribbon is composed of relatively flat beta-sheets rather than being the highly twisted, beta-coil structure previously suggested by analysis of globular protein folds. Comparison of the various fibril structures suggests that very small, local changes in beta-sheet twist are important in establishing the long range coiling of the protofilaments into fibrils of diverse morphology. PMID- 12093918 TI - Transepithelial transport of HIV-1 by M cells is receptor-mediated. AB - Human colon carcinoma Caco-2 cell monolayers undergo conversion into cells that share morphological and functional features of M cells when allowed to interact with B lymphocytes. A lymphotropic (X4) HIV-1 strain crosses M cell monolayers and infects underlying CD4(+) target cells. Transport requires both lactosyl cerebroside and CXCR4 receptors, which are expressed on the apical surface of Caco-2 and M cells. Antibodies specific for each receptor block transport. In contrast, a monotropic (R5) HIV-1 strain is unable to cross M cell monolayers and infect underlying monocytes, despite efficient transport of latex beads. Caco-2 and M cells do not express CCR5, but transfection of these cells with CCR5 cDNA restores transport of R5 virus, which demonstrates that HIV-1 transport across M cells is receptor-mediated. The follicle-associated epithelium covering human gut lymphoid follicles expresses CCR5, but not CXCR4, and lactosyl cerebroside, suggesting that HIV-1 infection may occur through M cells and enterocytes at these sites. PMID- 12093919 TI - Identification and characterization of CIA/ASF1 as an interactor of bromodomains associated with TFIID. AB - General transcription initiation factor IID (TFIID) plays a central and critical role in transcription initiation from both naked and chromatin templates. Although interaction between several DNA-binding proteins and TFIID were identified and well characterized, functional linkage between TFIID and chromatin factors has remained to be elucidated. Here we show the identification and characterization of human CIA/hASF1 (identified previously as a histone chaperone) as an interactor of two tandem bromodomain modules of human (h)TAF(II)250/CCG1, the largest subunit of TFIID. Although yeast (y)TAF(II)145, a homologue of hTAF(II)250/CCG1 in Saccharomyces cerevisiae, lacks bromodomains, glutathione S-transferase pull-down and immunoprecipitation assays revealed that Asf1p (antisilencing function 1), the counterpart of CIA in S. cerevisiae, interacts with Bdf1p (bromodomain factor 1), which is reported to serve as the missing bromodomain in yTAF(II)145. Furthermore, yeast strain lacking the BDF1 gene shows the Spt phenotype that is shown also by the ASF1 gene disruptant, and a double-knockout strain of both genes shows synthetic lethality, indicating that ASF1 genetically interacts with bromodomains associated with yTFIID. We also found that Asf1p coprecipitates with yTFIID subunits from yeast whole-cell extract, and overexpression of yTFIID subunits suppress the Spt phenotype caused by gene disruption of the ASF1. This study describes the functional linkage between TFIID and a histone chaperone. PMID- 12093920 TI - Crystal structure of human cytomegalovirus IL-10 bound to soluble human IL-10R1. AB - Human IL-10 (hIL-10) modulates critical immune and inflammatory responses by way of interactions with its high- (IL-10R1) and low-affinity (IL-10R2) cell surface receptors. Human cytomegalovirus exploits the IL-10 signaling pathway by expressing a functional viral IL-10 homolog (cmvIL-10), which shares only 27% sequence identity with hIL-10 yet signals through IL-10R1 and IL-10R2. To define the molecular basis of this virus-host interaction, we determined the 2.7-A crystal structure of cmvIL-10 bound to the extracellular fragment of IL-10R1 (sIL 10R1). The structure reveals cmvIL-10 forms a disulfide-linked homodimer that binds two sIL-10R1 molecules. Although cmvIL-10 and hIL-10 share similar intertwined topologies and sIL-10R1 binding sites, their respective interdomain angles differ by approximately 40 degrees. This difference results in a striking re-organization of the IL-10R1s in the putative cell surface complex. Solution binding studies show cmvIL-10 and hIL-10 share essentially identical affinities for sIL-10R1 whereas the Epstein-Barr virus IL-10 homolog (ebvIL-10), whose structure is highly similar to hIL-10, exhibits a approximately 20-fold reduction in sIL-10R1 affinity. Our results suggest cmvIL-10 and ebvIL-10 have evolved different molecular mechanisms to engage the IL-10 receptors that ultimately enhance the respective ability of their virus to escape immune detection. PMID- 12093921 TI - Vanadate-catalyzed photocleavage of the signature motif of an ATP-binding cassette (ABC) transporter. AB - The maltose transport complex of Escherichia coli, a member of the ATP-binding cassette (ABC) superfamily, is made up of two nucleotide-binding subunits, MalK(2), which hydrolyze ATP with positive cooperativity, and two transmembrane subunits, MalF and MalG. The ABC family is defined in part by the canonical signature motif LSGGQ whose exact function remains controversial. Taking advantage of the dual function of vanadate as a transition state analogue and as a photoactive chemical, we demonstrate that vanadate catalyzes the UV-dependent cleavage of the polypeptide backbone at both the LSGGQ motif and the nucleotide binding, or Walker A, motif when it is trapped in the nucleotide-binding site of the bacterial maltose transporter. This highly specific cleavage pattern indicates that residues in both motifs are immediately adjacent to ATP during hydrolysis, and are therefore likely to participate directly in ATP-binding and/or hydrolysis. Because the LSGGQ motif is too distant from the nucleotide in the structure of an ABC monomer for cleavage to occur, these data support a model in which the LSGGQ motif contacts the nucleotide across the interface of a MalK dimer, as seen in the crystal structure of Rad50. This architecture provides a basis for the cooperativity observed in the nucleotide-binding domains of ABC transporters and a function for this highly conserved family signature motif. PMID- 12093923 TI - Polyvalent cation receptor proteins (CaRs) are salinity sensors in fish. AB - To determine whether calcium polyvalent cation-sensing receptors (CaRs) are salinity sensors in fish, we used a homology-based cloning strategy to isolate a 4.1-kb cDNA encoding a 1,027-aa dogfish shark (Squalus acanthias) kidney CaR. Expression studies in human embryonic kidney cells reveal that shark kidney senses combinations of Ca(2+), Mg(2+), and Na(+) ions at concentrations present in seawater and kidney tubules. Shark kidney is expressed in multiple shark osmoregulatory organs, including specific tubules of the kidney, rectal gland, stomach, intestine, olfactory lamellae, gill, and brain. Reverse transcriptase PCR amplification using specific primers in two teleost fish, winter flounder (Pleuronectes americanus) and Atlantic salmon (Salmo salar), reveals a similar pattern of CaR tissue expression. Exposure of the lumen of winter flounder urinary bladder to the CaR agonists, Gd(3+) and neomycin, reversibly inhibit volume transport, which is important for euryhaline teleost survival in seawater. Within 24-72 hr after transfer of freshwater-adapted Atlantic salmon to seawater, there are increases in their plasma Ca(2+), Mg(2+), and Na(+) that likely serve as a signal for internal CaRs, i.e., brain, to sense alterations in salinity in the surrounding water. We conclude that CaRs act as salinity sensors in both teleost and elasmobranch fish. Their tissue expression patterns in fish provide insights into CaR functions in terrestrial animals including humans. PMID- 12093922 TI - The COXes of Danio: from mechanistic model to experimental therapeutics. PMID- 12093924 TI - Oxytocin induces differentiation of P19 embryonic stem cells to cardiomyocytes. AB - We recently discovered the existence of the oxytocin/oxytocin receptor (OT/OTR) system in the heart. Activation of cardiac OTR stimulates the release of atrial natriuretic peptide (ANP), which is involved in regulation of blood pressure and cell growth. Having observed elevated OT levels in the fetal and newborn heart at a stage of intense cardiomyocyte hyperplasia, we hypothesized a role for OT in cardiomyocyte differentiation. We used mouse P19 embryonic stem cells to substantiate this potential role. P19 cells give rise to the formation of cell derivatives of all germ layers. Treatment of P19 cell aggregates with dimethyl sulfoxide (DMSO) induces differentiation to cardiomyocytes. In this work, P19 cells were allowed to aggregate from day 0 to day 4 in the presence of 0.5% DMSO, 10(-7) M OT and/or 10(-7) M OT antagonist (OTA), and then cultured in the absence of these factors until day 14. OT alone stimulated the production of beating cell colonies in all 24 independently growing cultures by day 8 of the differentiation protocol, whereas the same result was obtained in cells induced by DMSO only after 12 days. Cells induced with OT exhibited increased ANP mRNA, had abundant mitochondria (i.e., they strongly absorbed rhodamine 123), and expressed sarcomeric myosin heavy chain and dihydropyridine receptor-alpha 1, confirming a cardiomyocyte phenotype. In addition, OT as well as DMSO increased OTR protein and OTR mRNA, and OTA completely inhibited the formation of cardiomyocytes in OT- and DMSO-supplemented cultures. These results suggest that the OT/OTR system plays an important role in cardiogenesis by promoting cardiomyocyte differentiation. PMID- 12093925 TI - Progress toward an ultimate proof of the prion hypothesis. PMID- 12093926 TI - Shooting from the hip: spatial control of signal release by intracellular waves. PMID- 12093927 TI - The search for new ways to treat obesity. PMID- 12093928 TI - Plant receptor kinases: systemin receptor identified. PMID- 12093929 TI - Exceptional haplotype variation in maize. PMID- 12093932 TI - Dysfunctional uterine bleeding. PMID- 12093933 TI - Managing stimulant medication for attention-deficit/hyperactivity disorder: an update. PMID- 12093930 TI - Effects of cerebral ischemia in mice deficient in Persephin. AB - Persephin (Pspn), a recently cloned member of the transforming growth factor-beta superfamily (TGF-beta) and glial cell line-derived neurotrophic factor (GDNF) subfamily, is distributed throughout the nervous system at extremely low levels and is thought to function as a survival factor for midbrain dopaminergic and spinal motor neurons in vivo. Here, we report that mice lacking Pspn by homologous recombination show normal development and behavior, but are hypersensitive to cerebral ischemia. A 300% increase in infarction volume was observed after middle cerebral artery occlusion. We find that glutamate-induced Ca(2+) influx, thought to be a major component of ischemic neuronal cell death, can be regulated directly by the Persephin protein (PSP) and that PSP can reduce hypoxia/reperfusion cell death in vitro. Neuronal cell death can be prevented or markedly attenuated by administration of recombinant human PSP in vivo before ischemia in both mouse and rat models. Taken together, these data indicate that PSP is a potent modulator of excitotoxicity in the central nervous system with pronounced neuroprotective activity. Our findings support the view that PSP signaling can exert an important control function in the context of stroke and glutamate-mediated neurotoxicity, and also suggest that future therapeutic approaches may involve this novel trophic protein. PMID- 12093934 TI - Nephrotic syndrome: pathogenesis and management. PMID- 12093935 TI - Index of suspicion. PMID- 12093936 TI - Question from the clinician: metatarsus adductus. PMID- 12093937 TI - Normal growth and revised growth charts. PMID- 12093938 TI - Effects on the fetus of maternal drugs during labor. PMID- 12093939 TI - Viral causes of diarrhea. PMID- 12093940 TI - Clinical characteristics of children with complicated pneumonia caused by Streptococcus pneumoniae. AB - OBJECTIVE: The frequency of children who are hospitalized with pneumococcal pneumonia complicated by necrosis, empyema/complicated parapneumonic effusion, and lung abscess seems to be increasing. The factors that contribute to this increase are unclear; therefore, the objective of this study was to describe and compare the relative frequency, clinical characteristics, and outcome of hospitalized children with complicated pneumonia with those of children with uncomplicated pneumonia caused by Streptococcus pneumoniae in the era of antibiotic resistance. METHODS: A multicenter, retrospective study of 8 children's hospitals in the United States was undertaken. A total of 368 children who were hospitalized with pneumococcal pneumonia identified from patients enrolled in the US Pediatric Multicenter Pneumococcal Surveillance Study over the period from September 1, 1993, to January 31, 2000 were studied. Demographic and clinical variables, antibiotic susceptibility, pneumococcal serotypes, antimicrobial therapy, and clinical outcome in hospitalized children with complicated versus uncomplicated pneumococcal pneumonia were measured. RESULTS: A total of 368 patients with pneumococcal pneumonia were identified. Of the 368 isolates, 47 (12.8%) were intermediate and 37 (10.1%) were resistant to penicillin; 18 (5%) were intermediate to ceftriaxone, and 9 (2.5%) were resistant to ceftriaxone. A total of 133 patients met the criteria for complicated pneumonia and had a chest tube placed; 56 of these patients subsequently underwent decortication. The proportion of hospitalized patients with complicated pneumococcal pneumonia increased progressively over the study period from 22.6% in 1994 to 53% in 1999. Patients with complicated disease were older (median age: 45 vs 27 months) and significantly more likely to be of white race and have chest pain on presentation compared with patients with uncomplicated disease. Patients who had complicated disease and underwent decortication were more likely to have pleural fluid lactate dehydrogenase levels of >7500 IU/L compared with those patients who had chest tube placement alone. Fifty-three percent of children who were > or =61 months of age and were hospitalized had complicated pneumonia. This group of children accounted overall for 42% of the patients with complicated pneumonia, 48.2% of the patients who subsequently underwent decortication, and 44% of the patients who had received a course of antibiotics before diagnosis. Pneumococcal serotypes 1, 6, 14, and 19 were the most prevalent serotypes causing disease, with serotype 1 causing 24.4% of the complicated cases versus 3.6% of the uncomplicated cases. Ninety-eight percent of the patients in both groups recovered from their pneumonia. Antibiotic resistance was not found to be more prevalent in those patients with complicated disease. CONCLUSIONS: The relative frequency of complicated disease in hospitalized children with pneumococcal pneumonia is increasing. Patients with complicated pneumococcal disease were older and significantly more likely to be of white race compared with those patients with uncomplicated disease. Pneumococcal serotype 1 caused significantly more disease in patients with complicated versus uncomplicated pneumonia. Patients with complicated disease were not more likely to be infected with an antibiotic-resistant isolate. PMID- 12093942 TI - Comparison of skin-to-skin (kangaroo) and traditional care: parenting outcomes and preterm infant development. AB - OBJECTIVE: To examine whether the kangaroo care (KC) intervention in premature infants affects parent-child interactions and infant development. METHODS: Seventy-three preterm infants who received KC in the neonatal intensive care unit were matched with 73 control infants who received standard incubator care for birth weight, gestational age (GA), medical severity, and demographics. At 37 weeks' GA, mother-infant interaction, maternal depression, and mother perceptions were examined. At 3 months' corrected age, infant temperament, maternal and paternal sensitivity, and the home environment (with the Home Observation for Measurement of the Environment [HOME]) were observed. At 6 months' corrected age, cognitive development was measured with the Bayley-II and mother-infant interaction was filmed. Seven clusters of outcomes were examined at 3 time periods: at 37 weeks' GA, mother-infant interaction and maternal perceptions; at 3-month, HOME mothers, HOME fathers, and infant temperament; at 6 months, cognitive development and mother-infant interaction. RESULTS: After KC, interactions were more positive at 37 weeks' GA: mothers showed more positive affect, touch, and adaptation to infant cues, and infants showed more alertness and less gaze aversion. Mothers reported less depression and perceived infants as less abnormal. At 3 months, mothers and fathers of KC infants were more sensitive and provided a better home environment. At 6 months, KC mothers were more sensitive and infants scored higher on the Bayley Mental Developmental Index (KC: mean: 96.39; controls: mean: 91.81) and the Psychomotor Developmental Index (KC: mean: 85.47; controls: mean: 80.53). CONCLUSIONS: KC had a significant positive impact on the infant's perceptual-cognitive and motor development and on the parenting process. We speculate that KC has both a direct impact on infant development by contributing to neurophysiological organization and an indirect effect by improving parental mood, perceptions, and interactive behavior. PMID- 12093941 TI - Tonsillectomy and adenotonsillectomy for recurrent throat infection in moderately affected children. AB - OBJECTIVE: In previous clinical trials involving children severely affected with recurrent throat infection (7 or more well-documented, clinically important, adequately treated episodes of throat infection in the preceding year, or 5 or more such episodes in each of the 2 preceding years, or 3 or more such episodes in each of the 3 preceding years), we found tonsillectomy efficacious in reducing the number and severity of subsequent episodes of throat infection for at least 2 years. The results seemed to warrant the election of tonsillectomy in children meeting the trials' stringent eligibility criteria but also provided support for nonsurgical management. We undertook the present trials to determine 1) whether tonsillectomy would afford equivalent benefit in children who were less severely affected than those in our earlier trials but who nonetheless had indications for tonsillectomy comparable to those in general use, and 2) whether, in such children, the addition of adenoidectomy would confer additional benefit. METHODS: We conducted 2 parallel randomized, controlled trials in the Ambulatory Care Center of Children's Hospital of Pittsburgh. To be eligible, children were required to have had a history of recurrent episodes of throat infection that met standards slightly less stringent than the standards used in our earlier trials regarding either the frequency of previous episodes or their clinical features or their degree of documentation, but not regarding >1 of those parameters. These reduced standards were nonetheless more stringent than those in current official guidelines, which list "3 or more infections of tonsils and/or adenoids per year despite adequate medical therapy" as an indication for tonsillectomy or adenotonsillectomy. Of 2174 children referred by physicians or parents, 373 met the current trials' eligibility criteria and 328 were enrolled. Of these, 177 children without obstructing adenoids or recurrent or persistent otitis media were randomized to either a tonsillectomy group, an adenotonsillectomy group, or a control group (the 3-way trial), and 151 children who had 1 or more such conditions were randomized to either an adenotonsillectomy group or a control group (the 2-way trial). Outcome measures were the occurrence of episodes of throat infection during the 3 years of follow-up; other, indirect measures of morbidity; and complications of surgery. RESULTS: By various measures, the incidence of throat infection was significantly lower in surgical groups than in corresponding control groups during each of the 3 follow-up years. However, even among control children, mean rates of moderate or severe episodes were low, ranging from 0.16 to 0.43 per year. Adenotonsillectomy was no more efficacious than tonsillectomy alone. Of 203 children treated with surgery, 16 (7.9%) had surgery-related complications of varying types and severity. CONCLUSIONS: The modest benefit conferred by tonsillectomy or adenotonsillectomy in children moderately affected with recurrent throat infection seems not to justify the inherent risks, morbidity, and cost of the operations. We conclude that, under ordinary circumstances, neither eligibility criteria such as those used for the present trials nor the criterion for surgery in current official guidelines are sufficiently stringent for use in clinical practice. PMID- 12093943 TI - Summer sunburn and sun exposure among US youths ages 11 to 18: national prevalence and associated factors. AB - OBJECTIVE: Epidemiologic evidence suggests that ultraviolet radiation (UVR), from sun exposure and sunburns during early life, is associated with increased risk of melanoma and other skin cancers in adulthood. The objective of this study was to gather national, population-based data on sun exposure and protection behaviors among US youths ages 11 to 18 to quantify the magnitude and patterns of sun exposure and establish baselines for continued surveillance. METHODS: A nationwide survey based on random-digit dialing and stratified, 2-stage national probability sampling was conducted of households with telephones in the 48 contiguous states of the United States during July through October of 1998; a population-based sample of 1192 youth ages 11 to 18 was studied to measure weighted prevalence estimates of summer sunburn and sun exposure and adjusted prevalence odds ratios and 95% confidence intervals from logistic regression analyses of sunburn. RESULTS: Among youths ages 11 to 18, 72% reported having had at least 1 summer sunburn, 30% reported at least 3, and 12% reported at least 5 sunburns. Before receiving their most serious summer sunburn, 39% of youths reported having applied sunscreen. Factors associated with increased odds of sunburn included greater sun sensitivity, white race, age younger than 16 years, more hours spent outdoors, and high desirability of a tan. CONCLUSIONS: Summer sunburn was the norm among US youths ages 11 to 18. The high frequency of sunscreen use during the sunburning episodes suggests the need to educate youth and parents better about proper use of these agents, as well as the importance of practicing other sun protection behaviors, such as wearing hats and protective clothing and avoiding the sun during peak exposure times. These data may serve as a baseline for tracking progress in skin cancer prevention efforts and will inform the crafting of future public health campaigns. PMID- 12093944 TI - Neutrophil activation in preterm infants who have respiratory distress syndrome. AB - OBJECTIVE: To study neutrophil activation in circulation as a sign of systemic inflammation in preterm infants with respiratory distress syndrome. METHODS: The study comprised very low birth weight preterm infants who had respiratory distress syndrome and required intubation and mechanical ventilation (n = 51), 1 day-old preterm infants who had no need for mechanical ventilation (n = 12), term infants (n = 47), and adult volunteers (n = 25). Neutrophil surface expression of CD11b was quantified with flow cytometry. RESULTS: In preterm infants with respiratory distress syndrome, neutrophil CD11b expression during the first day of life was higher than in cord blood (mean: 165 relative fluorescence units [RFU] [standard deviation [SD]: 53], n = 29 vs 83 RFU [SD: 21], n = 11; 95% confidence interval [CI] for difference: 59-106) or in preterm infants without mechanical ventilation (106 RFU [SD: 33], n = 12; 95% CI for difference: 17-90). CD11b expression decreased by age of 10 days. CD11b expression was lower in preterm cord than in term cord blood (95% CI for difference: 5-53). However, in preterm infants with respiratory distress syndrome aged 2 to 5 days, it was higher than in term infants of that age. CONCLUSIONS: The observations demonstrate an early transient postnatal neutrophil activation indicative of systemic inflammation that may contribute to the tissue injury in preterm infants with respiratory distress syndrome. PMID- 12093945 TI - Psychological outcomes in long-term survivors of childhood leukemia, Hodgkin's disease, and non-Hodgkin's lymphoma: a report from the Childhood Cancer Survivor Study. AB - OBJECTIVE: To evaluate and compare psychological outcomes in long-term survivors of pediatric leukemia, Hodgkin's disease, and non-Hodgkin's lymphoma and sibling controls. METHODS: Adult survivors of childhood leukemia, Hodgkin's disease, and non-Hodgkin's lymphoma (N = 5736) and sibling controls (N = 2565) were administered a long-term follow-up questionnaire allowing assessment of symptoms associated with depression and somatic distress. RESULTS: The majority of respondents in this study did not demonstrate symptomatology indicative of depression or somatic distress. Survivors, however, were significantly more likely than sibling controls to report symptoms of depression and somatic distress. Women were significantly more likely to indicate symptoms of depression and somatic distress than were men; however, this difference did not vary by survivor/sibling status. Similarly, socioeconomic (SES) variables predicted symptomatic levels of depression and somatic distress for both survivors and siblings, and these effects did not vary by survivor/sibling status. Among leukemia, Hodgkin's disease, and non-Hodgkin's lymphoma survivors, in addition to gender and SES, the only treatment variable that predicted scores indicating depressive symptomatology was exposure to intensive chemotherapy. Exposure to intensive chemotherapy also predicted scores indicative of somatic distress symptoms. No other medical variables, including diagnostic category, age at diagnosis, time since diagnosis, and duration of treatment, predicted symptomatic scores for depression and somatic distress. CONCLUSIONS: This large, sibling controlled, multisite study of young adult survivors of childhood leukemia, Hodgkin's disease, and non-Hodgkin's lymphoma found that survivors had significant increased risk for reporting symptoms of depression and somatic distress and that intensive chemotherapy added to this risk. However, being a cancer survivor did not compound the effects of gender and SES variables on the 2 outcomes measured. The ability of SES, gender, and treatment-related variables to predict psychological symptoms in this cohort of childhood survivors and sibling controls calls for future research into varied biological and psychosocial pathways by which cancer influences future psychosocial functioning. PMID- 12093946 TI - Postpartum discharge preferences of pediatricians: results from a national survey. AB - OBJECTIVE: To identify practice/physician characteristics that influence pediatricians' self-reported newborn discharge practices. METHODS: Of the pediatricians randomly surveyed through a national American Academy of Pediatrics periodic survey conducted in 2000, 490 were identified as routinely providing care for newborns in the nursery. These respondents rated the importance of 22 infant, maternal, and peripartum factors in determining readiness for nursery discharge on a 5-point Likert scale and reported their perceptions of optimal and minimal lengths of stay (LOS) for healthy term newborns. Importance of readiness factors was dichotomized as "high" (very important or important) versus "low" (neither, unimportant, or very unimportant). Relationships between pediatricians' responses and demographic information were explored using multivariate logistic regression. RESULTS: Most pediatricians (at least 81%) rated all 7 infant clinical factors (eg, stable, normal vital signs, successful feeding) as highly important determinants of discharge readiness. Women were 2 to 3 times more likely to rate maternal and peripartum factors such as maternal fatigue and stress, demonstration of maternal skills, breastfeeding knowledge or experience, adequacy of social support, maternal age <18 years, and low income/lack of financial resources as highly important. With respect to hospital LOS, women were twice as likely to identify an optimal LOS as >36 hours and a minimal LOS as >24 hours. Pediatricians in group settings were 3 times as likely as those in solo or 2-physician practices to advocate an optimal LOS >36 hours, and those with a high proportion of publicly insured or uninsured patients were less likely to identify an optimal LOS as >36 hours (odds ratio: 0.53). CONCLUSIONS: Female pediatricians report a more biopsychosocial approach to determining discharge readiness than their male counterparts, taking into account infant characteristics, maternal skills, and socioemotional issues that may affect the mother-infant pair's adjustment at home. The finding that those who provide care for the most financially vulnerable patients do not see the need for longer LOS is both surprising and of concern. The results support the need for a prospective critical examination of perinatal hospital discharge practices, such as the Pediatric Research in Office Settings Life Around Newborn Discharge Study. PMID- 12093947 TI - Assessment of a transcutaneous device in the evaluation of neonatal hyperbilirubinemia in a primarily Hispanic population. AB - OBJECTIVE: To compare estimates of serum bilirubin as determined by a transcutaneous device (BiliChek [BC]) with laboratory-measured total serum bilirubin (TSB) in a predominately Hispanic population in which a significant number of TSB values > or =15 mg/dL was anticipated. METHODS: A total of 248 Hispanic and 56 non-Hispanic neonates were studied. Transcutaneous measurements were performed by 1 investigator within 30 minutes of blood sampling for TSB; TSB was determined in a large clinical laboratory using the diazo Jendrassik-Grof with blank method. Agreement between BC and TSB determinations was assessed using Bland-Altman plots and the Bradley-Blackwood test. Interdevice comparisons were made among the BC devices. Predictive indices for TSB >10 mg/dL and >15 mg/dL were determined using various BC cutoff values. RESULTS: TSB was > or =15 mg/dL in 31% of the Hispanic neonates. BC generally tended to underestimate TSB determinations, and this trend was more pronounced when TSB was >10 mg/dL. Very high sensitivities were observed only when relatively low BC cutoff values were used to predict TSB >10 mg/dL or >15 mg/L. Relatively small numbers of infants had BC values in these low ranges. CONCLUSIONS: The tendency of BC to underestimate TSB limits its usefulness in neonates with relatively high TSB. In this population, most infants would have required additional evaluation to ensure that TSB was not >10 mg/dL or >15 mg/dL. It seems that the discrepancy between this study and previous studies of BC is related to our relatively large number of TSB values > or =15 mg/dL. PMID- 12093948 TI - Health care services utilization in children with obstructive sleep apnea syndrome. AB - OBJECTIVE: Little is known about the effects of obstructive sleep apnea syndrome (OSAS) on utilization of health care services in children. The present study compares medical service utilization by children with OSAS with that of healthy children. METHODS: A cross-sectional study of 287 consecutively recruited children (1-18 years) with OSAS and no concomitant diseases and a control group matched by age, gender, and geographic location was conducted at the Clalit Health Care Services clinic in the southern region of Israel. Children in the study group underwent nocturnal polysomnography (PSG) studies. The control group (N = 1149) was randomly selected from the Clalit Health Care Services database. PSG was performed for the OSAS patients. Indices of health care utilization 1 year before the PSG study were analyzed. RESULTS: A 226% increase in health care utilization was noted among children with OSAS. Children up to 5 years of age consumed more health care resources than children over 5 years. Children with OSAS consumed more health care services than the control group at all ages. The leading components of this high cost are utilization of more hospital days, drugs, and visits to the emergency department. The severity of the OSAS correlates directly to total annual costs and independently to age (beta = 0.19). CONCLUSIONS: Children with OSAS are heavy consumers of health care services 1 year before any specific evaluation and treatment for apnea. Early diagnosis and intervention may be cost-effective. PMID- 12093949 TI - Double-blind, randomized trial of long-chain polyunsaturated fatty acid supplementation in formula fed to preterm infants. AB - OBJECTIVE: We tested the hypothesis that balanced addition of long-chain polyunsaturated fatty acid (LCPUFA) to preterm formula during the first weeks of life would confer long-term neurodevelopmental advantage in a double-blind, randomized, controlled trial of preterm formula with and without preformed LCPUFA. METHODS: The participants were 195 formula-fed preterm infants (birth weight <1750 g, gestation <37 weeks) from 2 UK neonatal units and 88 breast milk fed infants. Main outcome measures were Bayley Mental Developmental Index (MDI) and Psychomotor Developmental Index (PDI) at 18 months and Knobloch, Passamanick and Sherrard's Developmental Screening Inventory at 9 months' corrected age. Safety outcome measures were anthropometry at 9 and 18 months, tolerance, infection, necrotizing enterocolitis, and death. RESULTS: There were no significant differences in developmental scores between randomized groups, although infants who were fed LCPUFA-supplemented formula showed a nonsignificant 2.6-point (0.25 standard deviation) advantage in MDI and PDI at 18 months, with a greater (nonsignificant) advantage (MDI: 4.5 points; PDI: 5.8 points) in infants below 30 weeks' gestation. LCPUFA-supplemented infants were shorter than control infants at 18 months (difference in length standard deviation score: 0.44; 95% confidence interval: 0.08-0.8). No other significant short- or long-term differences in safety outcomes were observed. Breastfed infants had significantly higher developmental scores at 9 and 18 months than both formula groups and were significantly heavier and longer at 18 months than LCPUFA-supplemented but not control infants. CONCLUSIONS: With the dose, duration, and preparation of LCPUFA used, efficacy was not demonstrated, although an advantage in later neurodevelopment cannot be excluded by global tests of development up to 18 months, particularly in infants below 30 weeks' gestation. The surprising effect of LCPUFA-supplemented formula on growth 18 months beyond the intervention period needs to be confirmed in other studies using similar supplementation strategies. Additional follow-up of this cohort is critical at an age when more specific tests of cognitive function are possible. PMID- 12093950 TI - Bedside diagnosis of influenzavirus infections in hospitalized children. AB - OBJECTIVE: For preventing nosocomial influenza infections and to facilitate prompt antiviral therapy, an accessible, rapid diagnostic method for influenzavirus is needed. We evaluated the performance of a lateral-flow immunoassay (QuickVue Influenza Test) completed at the bedside of hospitalized children during the influenza season. METHODS: All children who were evaluated at a large teaching hospital during the 1999 to 2000 influenza season were eligible if they were 1) younger than 19 years and hospitalized with respiratory symptoms or 2) younger than 3 years and hospitalized with fever. Each study child had 2 nasal swabs obtained--1 for influenzavirus culture and polymerase chain reaction (PCR) and the other for the QuickVue Influenza Test. The performance of the rapid diagnostic test was compared with the results of culture or PCR for influenza A or B. RESULTS: Of 303 eligible children, 233 (77%) were enrolled. In this population, 19 children had culture- and/or PCR-confirmed influenza A infection, prevalence of 8%. The QuickVue Influenza Test had a sensitivity of 74%, specificity of 98%, positive predictive value of 74%, and negative predictive value of 98%. CONCLUSIONS: Among children hospitalized with fever/respiratory symptoms during the influenza season, negative bedside QuickVue Influenza Tests indicated very low likelihood of influenza infection, whereas positive tests greatly increased the probability of influenza-associated illness. PMID- 12093951 TI - Differentiation between primary and secondary hypertension in children using ambulatory blood pressure monitoring. AB - OBJECTIVE: To determine whether ambulatory blood pressure monitoring (ABPM) can help differentiate children with secondary hypertension from those with primary hypertension. METHODS: Ninety-seven ABPM studies obtained from 85 children followed in a pediatric hypertension clinic were analyzed. Forty studies were performed in patients with primary hypertension, and 57 studies were performed in patients with secondary hypertension. Mean patient age was 13.8 +/- 3.5 [mean +/- standard deviation] years, range 4 to 19.7 years; patients with secondary hypertension were younger and had lower body mass index than patients with primary hypertension. RESULTS: Daytime diastolic and nocturnal systolic blood pressure (BP) loads, defined as the percentage of readings greater than a threshold value, were significantly greater in patients with secondary hypertension compared with patients with primary hypertension. A daytime diastolic BP load of > or =25% and/or a nocturnal systolic BP load of > or =50% was highly specific for secondary hypertension. CONCLUSIONS: Secondary hypertension in childhood is characterized by daytime diastolic BP elevation and nocturnal systolic BP elevation. This pattern of hypertension on ABPM may be a clue to underlying renal or other organ system pathology in children being evaluated for suspected hypertension and could help to identify children who require more detailed evaluation to determine the cause of their hypertension. PMID- 12093952 TI - Forced use treatment of childhood hemiparesis. AB - OBJECTIVE: Forced use, or constraint-induced, movement therapy has shown some efficacy in the rehabilitation of adults with chronic hemiparesis as a result of stroke. We used restraint of the unimpaired arm to ascertain whether this would improve function of the paretic arm in children with chronic (>1 year) hemiparesis. METHODS: Twelve hemiparetic treatment children (age 1-8 years) received a plaster cast on the unimpaired arm for 1 month; 13 hemiparetic control children did not. Peabody Developmental Motor Scales (PDMS) were performed on all treatment and control children immediately before and after casting and again 6 months later when controls crossed over to receive casts. Thus, PDMS were performed at entry, then 1 month, 6 months, and 7 months after entry. Any noted change in functional ability was also elicited by parental report. The frequency of visits to physical and occupational therapy was recorded. RESULTS: The 12 treatment (casted) children improved 12.6 PDMS points after 1 month of casting; the 13 control children improved 2.5 points. Improved PDMS scores persisted 6 months later when 7 treatment children returned. Similar results were obtained in the crossover when 10 control children received casts. Parental report corroborated improvement in casted children (22 of 22 parents) and its persistence at follow-up (21 of 22 parents). Receiving ongoing physical/occupational therapy did not seem to account for these results: control children received more (2.1 visits/wk) than treatment children (1.4 visits/wk). CONCLUSIONS: Forced use can be an effective rehabilitation technique in children with chronic hemiparesis. PMID- 12093953 TI - Early management of craniosynostosis using endoscopic-assisted strip craniectomies and cranial orthotic molding therapy. AB - OBJECTIVE: To assess the safety, efficacy, and results of the early treatment of infants with craniosynostosis using minimally invasive endoscopic strip craniectomies and postoperative helmet molding therapy. METHODS: A total of 100 patients with documented diagnosis of craniosynostosis were prospectively studied and treated with endoscopic strip craniectomies. A total of 106 stenosed sutures were operated on with the following distribution: 61 sagittal, 23 coronal, 18 metopic, and 4 lambdoid sutures. Sixty-three patients were treated under 16 weeks of age. After surgery, all patients were treated with custom-made molding helmets for up to 7 months. Follow-up ranged between 4 months and 50 months. RESULTS: All patients underwent the surgical procedures successfully and without complications. The mean surgical operative time was 52.7 minutes. The mean estimated blood loss was 26.2 mL; only 1 patient underwent intraoperative blood transfusion, and 10 patients had a non- life-threatening postoperative blood transfusion. All but 3 patients were discharged on the first postoperative day. There were no infections, dural sinus tears, cerebrospinal fluid leaks, or neurologic injuries, and there were no significant complications related to the use of helmet therapy. Most patients have achieved or are in the process of reaching normalization of their craniofacial deformities. CONCLUSIONS: The results indicate that the early treatment of craniosynostosis with minimally invasive endoscopic strip craniectomies is a safe, efficacious, and valuable therapeutic alternative to the current extensive surgical treatment modalities. The significantly less blood loss, need for blood transfusions, and length of stay and decreased costs make this procedure an excellent early option for treating infants who present with craniosynostosis. PMID- 12093954 TI - Urinary excretion of cross-linked N-telopeptides of type 1 collagen to assess bone resorption in infants from birth to 1 year of age. AB - OBJECTIVES: To evaluate noninvasively bone resorption in infants and more specifically, to assess the accuracy of urinary collagen type 1 cross-linked N telopeptide (NTX) excretion normalized to creatinine (NTX/Cr) in a spot urine sample as a reflection of daily NTX production in infants and to compute normative values for NTX excretion from birth to 1 year of age. METHODS: NTX/Cr values obtained from a single spot urine sample were compared with daily urinary NTX excretion and NTX/Cr obtained in 24-hour urine collected from 8 hospitalized infants. Normative values for NTX excretion were collated with a cross-sectional study in 70 healthy French infants (42 boys, 28 girls) aged 0 to 374 days (weight: 2700-11 340 g; length: 46-76.5 cm) and free of diseases or treatments that could influence growth, bone mineralization, or renal function. RESULTS: NTX/Cr values from single spot urine sample were significantly and linearly correlated with both daily NTX excretion (r = 0.783) and daily NTX/Cr (r = 0.952). In healthy infants, NTX excretion is low at birth, increases dramatically and significantly during the first 10 days of life, remains significantly elevated for approximately 3 months, and then decreases progressively to return to values similar to that observed at birth by 1 year of age. CONCLUSIONS: These data provide new insights regarding the use of spot urine analysis for assessing NTX excretion during the first year of life. The normative data demonstrate significant age-related variations in this marker, which probably reflect adaptation to extrauterine life and accelerated bone turnover in infancy and which should be considered for the interpretation of this noninvasive bone resorption marker in the clinical setting. PMID- 12093955 TI - Maternal bone lead as an independent risk factor for fetal neurotoxicity: a prospective study. AB - OBJECTIVE: A number of prospective studies have examined lead levels in umbilical cord blood at birth as predictors of infant mental development. Although several have found significant inverse associations, others have not. Measurement of lead levels in maternal bone, now recognized as the source of much fetal exposure, has the potential to serve as a better or complementary predictor of lead's effect on the fetus. Our objective was to compare lead levels in umbilical cord blood and maternal bone as independent predictors of infant mental development using a prospective design. METHODS: We recruited women who were giving birth at 3 maternity hospitals in Mexico City that serve a homogeneous middle-class community. Umbilical cord blood lead levels were measured by graphite furnace atomic absorption spectroscopy, and maternal lead levels in cortical (tibial) and trabecular (patellar) bone were measured within 4 weeks of giving birth using a 109-Cd K-x-ray fluorescence instrument. At 24 months of age, each infant was assessed using the Bayley Scales of Infant Development-II (Spanish Version). RESULTS: A total of 197 mother-infant pairs completed this portion of the study and had data on all variables of interest. After adjustment for other well-known determinants of infant neurodevelopment, including maternal age, IQ, and education; paternal education; marital status; breastfeeding duration; infant gender; and infant illness, lead levels in umbilical cord blood and trabecular bone were significantly, independently, and inversely associated with the Mental Development Index (MDI) scores of the Bayley Scale. In relation to the lowest quartile of trabecular bone lead, the second, third, and fourth quartiles were associated with 5.4-, 7.2-, and 6.5-point decrements in adjusted MDI scores. A 2 fold increase in cord blood lead level (eg, from 5 to 10 micro g/dL) was associated with a 3.1-point decrement in MDI score, which is comparable to the magnitude of effect seen in previous studies. CONCLUSION: Higher maternal trabecular bone lead levels constitute an independent risk factor for impaired mental development in infants at 24 months of age. This effect is probably attributable to mobilization of maternal bone lead stores, a phenomenon that may constitute a significant public health problem in view of the long residence time of lead in bone. PMID- 12093956 TI - CHARGE syndrome: a window of opportunity for audiologic intervention. AB - OBJECTIVE: To detail the clinical features of 22 new patients with a syndrome characterized by ocular coloboma, heart defects, atretic choanae, retarded growth or development, genital hypoplasia, and ear anomalies or hearing loss (CHARGE) seen in a tertiary academic medical center; compare auditory brainstem response (ABR) thresholds and behavioral hearing test results; identify a "window of opportunity" for audiologic intervention; review the literature regarding hearing results in CHARGE syndrome; and review the relationship between facial palsy and sensorineural hearing loss. METHODS: Clinical data were gathered to examine 1) the variety of hearing results, 2) the average age at the time of hearing loss identification in 22 children with CHARGE using electrophysiologic and behavioral test methods, 3) the usefulness of the ABR as an early indicator of hearing sensitivity for a select group composed of children from the present study and from an earlier report from the same institution, and 4) the value of congenital facial paralysis as a predictor of sensorineural hearing loss in CHARGE children seen in the authors' institution since 1983. RESULTS: All children had 4 or more acronymic features, including colobomatous defects or choanal atresia. Ear anomalies/hearing loss occurred at least as frequently as other primary features. A total of 81% of patients had hearing loss; in this subset, 1 child had a mild degree of loss, and the remaining children had moderate or greater losses. The average age at which ABR confirmed hearing status was 3.8 months, whereas for behavioral testing, that age was 24.7 months, a statistically significant difference. In a select group of 16 children, no statistical differences existed when comparing threshold results of early electrophysiologic testing with behavioral test findings obtained at a later date. Contingency analysis suggests that congenital facial paralysis and sensorineural hearing loss are related. CONCLUSIONS: Hearing loss is prevalent in children with CHARGE syndrome. Within a wide range of results exists a propensity for moderate or greater hearing loss in children with sensorineural or mixed impairments. Congenital facial palsy seems to be a valid statistical predictor of sensorineural hearing loss and can be a useful device in audiologic decision making. A "window of opportunity" for audiologic intervention exists in the first few months of life. Primary care providers are encouraged to recognize the need for immediate, early audiologic referral of their patients suspected to have CHARGE. Evaluation of hearing sensitivity during infancy and, when appropriate, provision of amplification are important components in the process of auditory habilitation. These efforts are in keeping with various professional guidelines that call for early detection of hearing loss and subsequent prompt intervention to minimize effects on infant development. PMID- 12093957 TI - Isoimmunization is unlikely to be the cause of hemolysis in ABO-incompatible but direct antiglobulin test-negative neonates. AB - OBJECTIVE: It is stated that the direct antiglobulin (Coombs') test (DAT) may be negative in ABO hemolytic disease of the newborn. Thus, significant jaundice in neonates who are A-B incompatible with their mothers but DAT test negative is often attributed to isoimmunization and another diagnosis is not sought. We wished to determine the rate of bilirubin production, as an objective measure of hemolysis, in 2 groups of DAT-negative neonates--ABO-compatible and ABO incompatible--and in DAT-positive ABO-incompatible neonates. METHODS: In consecutive, term, healthy newborns who were admitted to the general care nursery, we measured the level in parts per million (ppm) of end-tidal breath carbon monoxide (CO), corrected for inspired CO (ETCOc), an index of the rate of bilirubin production. We compared the levels in DAT-negative ABO-incompatible neonates with those in ABO-compatible neonates and with the levels in DAT positive ABO-incompatible neonates. Statistical analysis was performed using 2 sample t and chi(2) tests. RESULTS: There was no significant difference between the mean 12-hour ETCOc levels in DAT-negative ABO-incompatible neonates (n = 60, 2.2 +/- 0.6 ppm) versus DAT-negative ABO-compatible neonates (n = 171, 2.1 +/- 0.6 ppm), although there was a difference between the mean levels in DAT-positive ABO-incompatible neonates (n = 14, 3.4 +/- 1.8 ppm) and the DAT-negative groups. Four DAT-negative ABO-incompatible neonates had elevated ETCOc levels; in 2, we diagnosed a specific hematologic abnormality, namely, glucose-6-phosphate dehydrogenase deficiency in 1 and elliptocytosis in the other. CONCLUSION: In DAT negative newborns with significant jaundice or increased bilirubin production, even if ABO-incompatible, a cause other than isoimmunization should be sought. PMID- 12093958 TI - Changes in upper extremity position cause migration of peripherally inserted central catheters in neonates. AB - OBJECTIVE: The migration of peripherally inserted central catheters (PICCs) from the superior or inferior vena cava into the right atrium can pose a significant risk of lethal pericardial effusion and tamponade secondary to myocardial perforation. Arm movement has been reported to cause displacement of the catheter tip toward the heart and lead to ventricular tachycardia in adults. The objective of this study was to investigate whether adduction or abduction at the shoulder and flexion or extension at the elbow affect the position of PICCs placed via upper limb veins. We also hypothesized that arm movements can be used to reposition malpositioned catheters. METHODS: A total of 280 radiographs of 60 neonates with PICCs inserted via upper limb veins from July 2000 through June 2001 were reviewed. Differences in catheter tip position as a result of abduction versus adduction at the shoulder, flexion versus extension at the elbow, and combination changes in arm posture were determined by measurements in paired radiographs. Correction of malpositioned catheters was attempted in 10 patients by using arm movements without any alterations at the site of insertion. RESULTS: Arm movements were associated with significant displacement of catheters. Catheters that were placed via the basilic or axillary vein migrated toward the heart with adduction of the arm, whereas those that were placed via the cephalic vein moved away from the heart with adduction. Flexion of the elbow displaced catheters that were placed in the basilic or cephalic vein below the elbow toward the heart but did not have any effect on catheters that were placed via the axillary vein. For catheters that were placed in the basilic vein, simultaneous shoulder adduction and elbow flexion caused the greatest movement toward the heart (15.11 +/- 1.22 mm). We were able to reposition correctly inappropriately placed catheters in 9 of 10 patients by using arm movements. CONCLUSIONS: Arm movements significantly affect the position of the tip of the PICCs. Prevention of catheter migration into the right atrium requires radiographic determination of vein of insertion and monitoring of catheter tip position with upper extremity in position of maximum inward movement of catheter for that vein. Arm movements can be used to correct the malpositioned catheters. PMID- 12093959 TI - Pericardial effusion and tamponade in infants with central catheters. AB - OBJECTIVE: To describe the clinical presentation, cause, and outcome of central venous catheter (CVC)-related pericardial effusions (PCE) in infants. METHODS: A retrospective case review was conducted of CVC-related PCE at university and private neonatal intensive care units. Data from our cases were combined with published case reports and included clinical presentation and outcome; biochemical evaluation of pericardial fluid; days until diagnosis; cardiothoracic ratios; and CVC characteristics, insertion site, and tip placement site. RESULTS: In our cases, 6 different neonatology groups cared for 14 patients at 6 different hospitals in 2 cities. These data were combined with 47 cases reviewed from the literature. Pericardial fluid was obtained in 54 cases from the combined group and was described qualitatively as consistent with the infusate in 53 of 54 cases (98%). Biochemical analysis was performed in 37 cases, and in 36 of 37 cases (97%), the pericardial fluid was consistent with the infusate. The median gestational age at birth was 30.0 weeks (range: 23.5-42). The median time from CVC insertion to diagnosis was 3.0 days (range: 0.2-37; n = 59). Sudden cardiac collapse was reported in 37 cases (61%), and unexplained cardiorespiratory instability was reported in 22 cases (36%). The CVC tip was last reported within the pericardial reflections on chest radiograph in 56 cases (92%) at the time of PCE diagnosis. The mean cardiothoracic ratio increased 17% (n = 14). No patients died among our cases. Among the reviewed cases, 45% mortality was reported. For the combined group, mortality was 34%. For the combined group, mortality was 8% (3 of 37) in the patients who underwent pericardiocentesis versus 75% (18 of 24) for the patients who did not. In 21 patients, the catheter was withdrawn and remained in use. Survivors and nonsurvivors had comparable gestational age at birth, birth weight, days to PCE diagnosis, and day of life of PCE symptoms/diagnosis. Access site, catheter type, and catheter size were not associated with mortality. An association between larger catheters and shorter time to PCE may be present. Access site and catheter type were not associated with time to PCE. Autopsy specimens reported 6 cases of myocardial necrosis/thrombus formation, 9 cases of perforation without myocardial necrosis/thrombus formation, and 2 cases in which both were reported. CONCLUSIONS: The pericardial fluid found in CVC-associated PCE is consistent with the infusate. We speculate that there are several mechanisms, ranging from frank perforation that seals spontaneously to CVC tip adhesion to the myocardium with diffusion into the pericardial space. Routine radiography should be performed, and the CVC tip should be readily identifiable. The CVC tip should remain outside the cardiac silhouette but still within the vena cavae (approximately 1 cm outside the cardiac silhouette in premature infants and 2 cm in term infants). A change in cardiothoracic ratio may be diagnostic of a PCE, and pericardiocentesis is associated with significantly reduced mortality. Increased awareness of this complication may decrease the mortality associated with CVC-related PCE. PMID- 12093961 TI - Continuous professional development: raising the bar for pediatricians. PMID- 12093960 TI - Trends in mortality and morbidity for very low birth weight infants, 1991-1999. AB - BACKGROUND: Medical care for very low birth weight (VLBW) infants and their mothers has changed dramatically during the 1990s, yet it is unclear how these changes have affected mortality and morbidity. OBJECTIVE: We used the Vermont Oxford Network Database to identify trends in clinical practice and patient outcomes for VLBW infants born from 1991 to 1999. METHODS: Logistic regression was used to evaluate temporal trends in practices and outcomes while adjusting for patient characteristics and accounting for clustering of cases within hospitals. RESULTS: There were 118 448 infants 501 to 1500 g from 362 neonatal intensive care units enrolled in the Network Database from 1991 to 1999. Prenatal care, cesarean section, multiple births, antenatal steroids, and 1-minute Apgar scores increased during this period, as did the use of nasal continuous positive airway pressure, high-frequency ventilation, surfactant, and postnatal steroids. The proportion of white infants decreased; the proportions of Hispanic infants and those of other races increased. The crude and adjusted rates of mortality, pneumothorax, intraventricular hemorrhage (IVH), and severe IVH declined from 1991 to 1995, whereas from 1995 to 1999, the rates of mortality, IVH, and severe IVH did not change significantly, and pneumothorax increased. CONCLUSIONS: There have been major changes in both obstetric and neonatal care during the 1990s. These changes were associated with decreases in mortality and morbidity for VLBW infants during the first half of the decade. However, since 1995, no additional improvements in mortality or morbidity have been seen, ending a decades-long trend of improving outcomes for these infants. PMID- 12093962 TI - In times of crisis, what's a pediatrician to do? PMID- 12093963 TI - The misshapen head. PMID- 12093964 TI - Coding for mental health and behavioral problems: the arcane elevated to the ranks of the scientific. PMID- 12093965 TI - The use of recombinant activated factor VII to control bleeding in a preterm infant undergoing exploratory laparotomy. AB - The case of a preterm infant weighing 1120 g who successfully received recombinant activated factor VII (rFVIIa) without complication for control of a life-threatening bleeding event resulting from a ruptured umbilical artery is reported. After performing an exploratory laparotomy at 27 hours of age, hemorrhage from the surgical wound and various sites persisted. By 63 hours of age, the infant had received a total of 192 mL (171 mL/kg) of packed red blood cells, 115 mL (103 mL/kg) of fresh frozen plasma, 8 mL of cryoprecipitate, and 75 mL (67 mL/kg) of platelet concentrate without stabilization. Hemorrhage ceased after 2 doses of 40 microg/kg/dose recombinant activated factor VII given at 63 and 70 hours of age, with subsequent stabilization of the hematocrit and without need for additional transfusion therapy. PMID- 12093966 TI - Recurrent nursemaid's elbow (annular ligament displacement) treatment via telephone. AB - Annular ligament displacement (ALD)--also termed radial head subluxation, nursemaid's elbow, or pulled elbow--can be successfully diagnosed and treated over the telephone by properly trained medical professionals instructing nonmedical caretakers. Two case reports of successful ALD reduction via telephone are described. The pathology of ALD and techniques for its treatment are reviewed, and guidelines are given. The rationale for the introduction of the new term annular ligament displacement as well as areas for additional investigation are discussed. To our knowledge, this is the first published account of ALD reduction via telephone. PMID- 12093967 TI - The "Ospedale degli Innocenti" and the "bambino" of the American Academy of Pediatrics. PMID- 12093968 TI - Uses of drugs not described in the package insert (off-label uses). AB - New regulatory initiatives have been designed to ensure that new drugs and biologicals include adequate pediatric labeling for the claimed indications at the time of, or soon after, approval. However, because such labeling may not immediately be available, off-label use (or use that is not included in the approved label) of therapeutic agents is likely to remain common in the practice of pediatrics. This policy statement was written to address questions practitioners have regarding off-label use. The purpose of off-label use is to benefit the individual patient. Practitioners may use their professional judgment to determine these uses. Practitioners should understand that the Food and Drug Administration does not regulate off-label use. PMID- 12093969 TI - The medical home. AB - The American Academy of Pediatrics proposed a definition of the medical home in a 1992 policy statement. Efforts to establish medical homes for all children have encountered many challenges, including the existence of multiple interpretations of the "medical home" concept and the lack of adequate reimbursement for services provided by physicians caring for children in a medical home. This new policy statement contains an expanded and more comprehensive interpretation of the concept and an operational definition of the medical home. PMID- 12093970 TI - Guidelines for referral to pediatric surgical specialists. PMID- 12093971 TI - 'Sinusitis'? PMID- 12093973 TI - The dummies' guide to risk-benefit analysis of vaccines. PMID- 12093974 TI - Politics and reading. PMID- 12093975 TI - Amoxicillin dosage. PMID- 12093976 TI - Blood lead levels. PMID- 12093977 TI - Epidural analgesia and fever. PMID- 12093978 TI - Acetaminophen toxicity in children. PMID- 12093979 TI - More about heliox and bronchiolitis. PMID- 12093980 TI - Conflicting views in poison treatment. PMID- 12093981 TI - Premature conclusions on postnatal steroid effects. PMID- 12093982 TI - Pagers combined with telephones improve successful follow-up from a pediatric emergency department. AB - OBJECTIVE: To determine whether there is a significant difference in initial successful contact when attempting follow-up of patients discharged from a pediatric emergency department (ED) using either pagers or the telephone. In addition, to evaluate whether the combination of both pager and telephone follow up is more successful than telephone follow-up alone when confined to a 2-hour period. DESIGN AND SETTING: A prospective comparison of intervention and control groups taken from convenience samples of patients seen in an ED of an urban, tertiary care children's hospital. PARTICIPANTS: One hundred eighty-five patients whose families had access to both a pager and a telephone (intervention group) and 112 patients whose families had access to only a telephone (control group) were enrolled. INTERVENTION: Before discharging the patient from the ED, the investigators verified a pager number and/or a best contact telephone number for a follow-up telephone call. Participants were divided into 2 groups. The intervention group consisted of caretakers with both pagers and telephones. The control group consisted of caretakers with only telephones. On even calendar days after ED visits, intervention group caretakers were paged initially from 11:00 AM to 11:59 AM. On odd calendar days, intervention group caretakers were telephoned initially from 11:00 AM to 11:59 AM. Successful contact was defined as communication with a family member or guardian over the age of 18. If the caretaker spoke only Spanish, a translator was used. In a crossover design with the intervention group, if contact was unsuccessful after 1 hour, the alternative mode of communication was used at noon. Control group caretakers were telephoned from 11:00 AM to 11:59 AM. If telephone contact was unsuccessful, they were called again 1 hour later. RESULTS: Two hundred forty-six (36%) of 685 of the convenience sample of caretakers had access to both a pager and a telephone. Fifty-two (55%) of 94 intervention caretakers were contacted initially using pagers versus 47 (52%) of 91 intervention caretakers contacted initially by telephones. Overall successful contact of intervention caretakers (telephones and pagers) was 145 (78%) of 185 when confined to a 2-hour time period. In contrast, overall successful contact of control caretakers was 68 (61%) of 112 when confined to a 2-hour time period. Successful contact was greater with the intervention caretakers than with control caretakers (78% vs 61%). CONCLUSIONS: No significant difference in successful contact was seen whether paged or telephoned initially. The combination of both pagers and telephone follow-up was more successful than telephone follow-up alone when confined between 11:00 AM to 1:00 PM. PMID- 12093983 TI - Efficacy and safety of pimecrolimus cream in the long-term management of atopic dermatitis in children. AB - OBJECTIVE: Pimecrolimus cream (SDZ ASM 981), a nonsteroid inhibitor of inflammatory cytokines, is effective in atopic dermatitis (AD). We assessed whether early treatment of AD signs/symptoms with pimecrolimus could influence long-term outcome by preventing disease flares. METHODS: Early intervention with pimecrolimus was compared with a conventional AD treatment strategy (ie, emollients and topical corticosteroids). In this 1-year, controlled, double-blind study, 713 AD patients (2-17 years) were randomized 2:1 to a pimecrolimus-based or conventional regimen. Both groups used emollients for dry skin. Early AD signs/symptoms were treated with pimecrolimus cream or, in the conventional treatment group, vehicle to prevent progression to flares. If flares occurred, moderately potent topical corticosteroids were mandated. The primary efficacy endpoint was ranked flares at 6 months. Safety was monitored clinically, and a skin recall-antigen test was performed at study completion. RESULTS: BASELINE CHARACTERISTICS OF THE PATIENTS: The mean age for both groups was approximately 8 years, and the majority of patients had moderate disease at baseline. PATIENT FOLLOW-UP AND EXPOSURE TO STUDY MEDICATION: The mean duration of follow-up (+/ standard error) was 303.7 (+/-5.30) days in the pimecrolimus group and 235.2 (+/ 9.40) days in the control group. The discontinuation rate was significantly higher in the control group than in the pimecrolimus group (51.5% vs 31.6% at 12 months), and proportionately more patients with severe or very severe disease discontinued in the control group. The main reason for the higher discontinuation rate in the control group was unsatisfactory therapeutic effect (30.4% vs 12.4%). This resulted in a substantially higher mean number of study medication treatment days in the pimecrolimus group compared with the control group: 211.9 (69.8% of study days) versus 156.0 (66.3% of study days). Of those patients who completed 12 months on study, 14.2% and 7.0% of patients in the pimecrolimus and vehicle groups, respectively, used study medication continuously. EFFICACY: Patients in the pimecrolimus group experienced significantly fewer AD flares than those in the control group, according to the primary efficacy analysis on ranked flares of AD (Van Elteren test). The proportion of patients who completed 6 or 12 months with no flares was approximately twice as high in the pimecrolimus group compared with control (61.0% vs 34.2% at 6 months; 50.8% vs 28.3% at 12 months). Fewer flares were observed in the pimecrolimus group regardless of baseline disease severity, so even severe patients derived benefit from the treatment. The analysis of time to first flare showed that treatment with pimecrolimus was associated with a significantly longer flare-free period (log- rank test). Covariate analysis indicated a statistically significant effect on time to first flare of baseline Eczema Area and Severity Index score, and whether patients had "severe" or "very severe" disease at baseline according to the Investigators' Global Assessment, although patients in all baseline disease severity subgroups benefited from treatment. Age had no significant effect. Fewer patients in the pimecrolimus group required topical corticosteroid therapy compared with control (35.0% vs 62.9% at 6 months; 42.6% vs 68.4% at 12 months), and patients in the pimecrolimus group spent fewer days on topical corticosteroid therapy (57.4% vs 31.6% [pimecrolimus vs control, respectively] spent 0 days on topical corticosteroid therapy, 17.1% vs 27.5% 1-14 days, and 25.5% vs 41.0% >14 days over the 12 months of the study). This steroid-sparing effect of pimecrolimus was evident despite pimecrolimus-treated patients being on study longer than patients in the control group. The average proportion of study days spent on second-line corticosteroids was 4.08% in the pimecrolimus group and 9.10% in the control group. Analysis of Eczema Area and Severity Index over time showed significantly lower median scores, thus indicating better disease control in the pimecrolimus group compared with the control group. Similar results were obtained from analysis of the Investigators' Global Assessment (not shown). The treatment groups were well balanced with respect to the number of patients using antihistamines during the study (57.2% vs 62.9%, pimecrolimus vs control, respectively). SAFETY: There were no appreciable differences between treatment groups in the overall incidence of adverse events. The most frequent adverse events were common childhood infections and ailments, including nasopharyngitis, headache, and cough. The incidence of suspected drug-related adverse events was not significantly different in the pimecrolimus group (24.7% vs 18.7%- pimecrolimus vs control), and the incidence of serious adverse events was low (8.3% vs 5.2%--pimecrolimus vs control). Life-table analysis of incidence of adverse events revealed no significant differences between the treatment groups, except for cough. Local tolerability was good in both treatment groups. The most common application site reaction reported was sensation of burning (10.5% vs 9.3% -pimecrolimus vs control). There were no major differences between treatment groups in the duration or severity of application site reactions, most of which were mild-to-moderate and transient, occurring within the first week of treatment. Skin infections were reported in both groups. There were no between group differences in the life-table analysis of time to first occurrence of bacterial skin infections nor in the adjusted incidence of bacterial skin infections. Although there were no significant differences between treatment groups in the incidence of individual viral skin infections, the incidence of grouped viral skin infections (12.4% vs 6.3%--pimecrolimus vs control) showed a slightly higher incidence in the pimecrolimus group. Laboratory values and vital signs showed no significant between-group differences. There were no significant differences between treatment groups in response to recall antigens in those patients who remained on study for 12 months. CONCLUSIONS: Treatment of early AD signs/symptoms with pimecrolimus was effective in preventing progression to flares in more than half the patients, reducing or eliminating the need for topical corticosteroids. The benefits were consistently seen at 6 months across important disease severity subgroups and with respect to the various predefined efficacy endpoints. Furthermore, these benefits were sustained for 12 months, providing evidence that long-term treatment with pimecrolimus leads to better control of AD. Treatment with pimecrolimus was well tolerated and was not associated with clinically relevant adverse events compared with the conventional treatment group. The results reported here offer the prospect of effective long term management of AD with reduced need for topical corticosteroids. PMID- 12093984 TI - Extremity pain and refusal to walk in children with invasive meningococcal disease. AB - OBJECTIVE: Early recognition of invasive meningococcal disease in children may be difficult. Extremity pain and refusal to walk (extremity symptoms) are uncommonly mentioned as clinical findings in children who present with this disease. We sought to determine 1) the frequency of extremity symptoms as part of the clinical presentation in children with invasive meningococcal disease and 2) whether these symptoms help identify children with otherwise unsuspected meningococcal disease. METHODS: We reviewed the medical records of patients who were younger than 20 years and had invasive meningococcal disease from 1985 to 1996 at 3 pediatric referral centers. Children with extremity symptoms were identified and described. We compared clinical and laboratory findings and frequency of adverse outcomes between these children and those with invasive meningococcal disease without extremity symptoms. RESULTS: We identified 274 children with invasive meningococcal disease, 45 (16%) of whom had either history or physical examination evidence of extremity pain (31) or refusal to walk (14) as part of their clinical presentations. Five of the 45 patients had arthritis at the time of presentation. Patients with extremity symptoms at presentation were significantly older (77.9 +/- 62.2 vs 44.0 +/- 56.9 months), had lower temperatures (38.8 +/- 1.2 degrees C vs 39.2 +/- 1.2 degrees C), and had higher band counts (28.2 +/- 15.2% vs 18.1 +/- 12.4%) than did patients without extremity symptoms. There were no significant differences, however, between groups with regard to rash, white blood cell counts, coagulation parameters, prevalence of meningitis, or adverse outcomes. Seventy-three (27%) of the 274 patients had unsuspected disease, and 5 (7%) of these had extremity symptoms at the time of diagnosis. CONCLUSIONS: Sixteen percent of children with invasive meningococcal disease have extremity symptoms at the time of diagnosis. These symptoms may help to identify some patients with otherwise unsuspected invasive meningococcal disease. PMID- 12093985 TI - No evidence of autoimmunity in 6-year-old children immunized at birth with recombinant hepatitis B vaccine. AB - OBJECTIVES: Taking into account that genetic predisposition, marked by human leukocyte antigen (HLA) class I and II genes, augments the probability of developing an autoimmune disorder after a triggering vaccination, as largely debated, we investigated the frequency of autoantibody production after recombinant hepatitis B vaccine (rHBv) in 6-year-old children immunized at birth to evaluate an association between autoimmune disorders and hepatitis B virus vaccination. METHODS: We investigated the presence of autoantibodies in 210 6 year-old children who were immunized at birth with rHBv: 200 showed anti hepatitis B surface antigen concentrations > or =10 mUI/mL at seroconversion (responders), and 10 were nonresponders. Data were compared with those obtained in 109 unvaccinated children. All participants were screened for the presence of antinuclear antibodies (ANAs), anti-DNA, antimitochondrial, anti-liver/kidney microsomal, antireticulin, anti-smooth muscle (SMA), and antiribosomal antibodies. All participants were also screened for the presence of antithyroid antibodies, such as antithyroglobulin and antiperoxidase, and for antibodies found in type 1 diabetes, such as tyrosine phosphatase (IA-2A) and glutamic acid decarboxylase (GADA). HLA typing was extended to all 10 nonresponders. RESULTS: Autoantibodies were found in 16 of the 200 responders: ANAs were found in 12 (6%), smooth muscle antibodies were found in 4 (2.0%), and antireticulin antibodies and endomysial antibodies were found in 1 girl with ANAs. Antithyroid antibodies, IA-2A, and GADA were not present in any of the participants. No significant difference was found in the frequency of autoantibodies between vaccinated and control children. Three of the 10 nonresponder children were SMA positive (30% vs 2% of responders); they also carried the supratype HLA C4AQ0,DRB1*0301,DQB1*02. A family history for autoimmune disorders was present in 3 (18%; 95% confidence interval [CI]: 4.0%-45.6%) of the 16 responder infants with autoantibodies, in 15 (8.4%; 95% CI: 4.6%-13.1%) of responder children without autoantibodies, and in 1 (10%) of the 10 nonsreponder children. CONCLUSIONS: From our data, vaccination with rHBv given during the neonatal period does not seem to increase autoantibody production in a 6-year-old children. Autoantibodies, referred to as natural autoantibodies, can be found in healthy participants, but their significance is unclear. These autoantibodies often cross-react with bacteria or tumor antigens, suggesting their importance in innate immunity. It has been demonstrated in an animal model that self-antigen can promote B-cell accumulation, and that a significant proportion of natural autoantibodies is the product of this self-antigen- dependent process. Consequently, it has been speculated that self-antigens play a positive role in recruiting B cells as a part of innate immunity, but this process carries a potential risk for unregulated growth. Spreading of the immune response is a common theme in organ-specific and systemis autoimmune diseases, and this could be initiated by exogenous agents, in genetically susceptible hosts, owing to molecular mimicry of natural antigen. Moreover, 3 (18%) of the 16 children who had autoantibodies had a family history of autoimmume diseases. Thus, it is apparent that susceptibility to autoimmunity is determined by genetic factors rather than by vaccine challenge. Among all the children considered, only 1 girl (0.5%) developed celiac disease, reflecting the prevalence described in the literature. GADA and IA-2A were not found in our children; this observation is in agreement with data showing that type 1 diabetes risk may not be altered by vaccinations administered during childhood. On the contrary, a high frequency (30%) of autoantibodies, in particular SMA, was observed in the nonresponder children. The 3 SMA-positive children carried the HLA-C4Q0,DRB1*0301,DQB1*02 haplotype, a well-known predisposing factor for autoimmune disorders. On the other hand, the presence of autoantibodies to smooth muscle is known to be common in hepatitis B infection, and, it has been shown that cross-reactive immunity targeting homologous self-protein may partly account for autoantibody production. Although hepatitis B vaccination given during the neonatal period does not increase autoantibody production in 6-year-old immunized children, we deem useful a more prolonged follow-up for these nonresponder children carrying certain HLA haplotypes (such as C4AQ0,DRB1*0301,DQB1*02), particularly because most autoimmune diseases do not develop until later in life. PMID- 12093986 TI - Bone density and metabolism in children and adolescents with moderate to severe cerebral palsy. AB - OBJECTIVES: Diminished bone density and a propensity to fracture with minimal trauma are common in children and adolescents with moderate to severe cerebral palsy (CP). The purpose of this study was to provide a detailed evaluation of bone mineral density (BMD) and metabolism in this population and to assess the relationship of these measures to multiple other clinical, growth, and nutrition variables. METHODS: The study group consisted of 117 subjects ages 2 to 19 years (mean: 9.7 years) with moderate to severe CP as defined by the Gross Motor Functional Classification scale. Population-based sampling was used to recruit 62 of the participants, which allows for estimations of prevalence. The remaining 55 subjects were a convenience sampling from both hospital- and school-based sources. The evaluation included measures of BMD, a detailed anthropometric assessment of growth and nutritional status, medical and surgical history, the Child Health Status Questionnaire, and multiple serum analyses. BMD was measured in the distal femur, a site specifically developed for use in this contracted population, and the lumbar spine. BMD measures were converted to age and gender normalized z scores based on our own previously published control series (n > 250). RESULTS: Osteopenia (BMD z score <-2.0) was found in the femur of 77% of the population-based cohort and in 97% of all study participants who were unable to stand and were older than 9 years. BMD was not as low in the lumbar spine (population-based cohort mean +/- standard error z score: -1.8 +/- 0.1) as in the distal femur (mean z score: -3.1 +/- 0.2). Fractures had occurred in 26% of the children who were older than 10 years. Multiple clinical and nutritional variables correlated with BMD z scores, but interpretation of these findings is complicated by covariance among variables. In stepwise regression analyses, it was found that severity of neurologic impairment as graded by Gross Motor Functional Classification level, increasing difficulty feeding the child, use of anticonvulsants, and lower triceps skinfold z scores (in decreasing order of importance) all independently contribute to lower BMD z scores in the femur. CONCLUSIONS: Low BMD is prevalent in children with moderate to severe CP and is associated with significant fracture risk. The underlying pathophysiology is complex, with multiple factors contributing to the problem and significant variation between different regions of the skeleton. PMID- 12093987 TI - Relationship of adherence to pediatric asthma morbidity among inner-city children. AB - OBJECTIVES: Morbidity from asthma among children is one of the most important US health concerns. This study examines the relationship of baseline nonadherence to subsequent asthma morbidity among inner-city children. METHODS: A multisite, prospective, longitudinal panel study was conducted of 1199 children who were aged 4 to 9 years and had asthma and their caregivers, most of whom were parents, in emergency departments and clinics at 8 research centers in 7 US metropolitan inner-city areas. Nine morbidity indicators were collected at 3, 6, and 9 months after baseline, including hospitalizations, unscheduled visits, days of wheeze/cough, and days of reduced activities. RESULTS: Children whose caregivers scored high on a new measure, Admitted Nonadherence, experienced significantly worse morbidity on 8 of the 9 measures. Children who scored high on a new Risk for Nonadherence measure experienced significantly worse morbidity on all 9 morbidity measures. Multiple and logistic regressions found that the adherence measures had independent significant effects on morbidity. Combining the measures improved estimates of morbidity: children whose caregivers were poor on either adherence measure had worse morbidity than those with good adherence on both, eg, rate of hospitalization was twice as high, they missed more than twice as much school, had poorer overall functioning, and experienced more days of wheezing and more restricted days of activity. CONCLUSIONS: Risk for Nonadherence and Admitted Nonadherence independently and jointly predicted subsequent asthma morbidity. Targeting risks for nonadherence may be an effective intervention strategy. Most risks can be controlled by physicians through reducing the complexity of asthma regimens, communicating effectively with caregivers about medication use, and correcting family misconceptions about asthma medication side effects. PMID- 12093988 TI - Fecal elastase-1 is superior to fecal chymotrypsin in the assessment of pancreatic involvement in cystic fibrosis. AB - OBJECTIVE: Exocrine pancreatic function in patients with cystic fibrosis (CF) can be evaluated by direct and indirect tests. In pediatric patients, indirect tests are preferred because of their less invasive character, especially in CF patients with respiratory disease. Fecal tests are noninvasive and have been shown to have a high sensitivity and specificity. However, there is no comparative study in CF patients. Therefore, the aim of the present study was to compare the sensitivity and the specificity of the fecal elastase-1 (E1) test with the fecal chymotrypsin (ChT) test in a large cohort of CF patients and healthy subjects (HS). DESIGN: One hundred twenty-three CF patients and 105 HS were evaluated. In all subjects, E1 concentration and ChT activity were measured. In the CF group, fecal fat excretion was also determined. The sensitivity and specificity of the fecal E1 test and ChT test were compared. RESULTS: With a cutoff level of 3 U/g, ChT specificity in HS was similar to that of E1, but E1 sensitivity in CF patients was significantly higher (90.2% vs 81.3%). With a cutoff level of 6 U/g, ChT and E1 sensitivity in CF patients was identical, but E1 specificity in HS was again significantly higher (98.1% vs 90.5%). In all CF patients with severe steatorrhea (>15 g/d), E1 concentrations were abnormal and ChT activity was lower than 3 U/g. In contrast, in pancreatic-sufficient patients and patients with mild steatorrhea (< or =15 g/d), the E1 sensitivity was significantly higher compared with ChT (69.2% vs 41.0%). CONCLUSIONS: The fecal E1 test is superior to fecal ChT determination in the assessment of CF pancreatic involvement in pancreatic sufficient patients and those patients with mild steatorrhea. PMID- 12093989 TI - Coding of pediatric behavioral and mental disorders. AB - BACKGROUND: In response to changing reimbursement and other pressures in the health care environment, many physicians have reported the use of alternate coding to substitute for certain clinical diagnoses. However, very little information is available on how physicians who care for children approach diagnosis and coding dilemmas for behavioral and mental disorders, which often present unique additional challenges. OBJECTIVE: Our study sought to describe the frequency of alternate coding, different approaches to coding, and attitudes toward diagnosis and coding practices by physician specialty. METHODS: We conducted a mail survey of 1492 physicians--497 developmental/behavioral pediatricians (DBP), 500 pediatricians (PED), and 495 child and adolescent psychiatrists (PSY). The main outcomes were survey items on frequency of alternate coding (never, rarely, monthly, weekly, daily), use of different coding strategies (use of somatic symptoms, modifiers, and substitution with other terms), and attitudes on coding practices (Likert scales of agreement). We analyzed outcomes by physician specialty and demographics using Pearson's chi2 and multivariate logistic regression. RESULTS: Overall response rate was 62% (787 of 1269 eligible physicians). The majority of physicians had used an alternate code (DBP 83%, PED 68%, PSY 58%), and many respondents reported monthly-daily alternate coding (DBP 60%, PED 36%, PSY 27%). Physicians used multiple approaches to diagnosis and a variety of coding options, which varied by physician specialty. Financial issues were commonly cited reasons for alternate coding- both to obtain patient services and to receive physician reimbursement. However, challenges of diagnostic classification and coding subthreshold symptoms were cited as frequently as reimbursement issues. Stigmatization, confidentiality, and parental acceptance were mentioned, but reported less frequently. Very few practices and providers have organized administrative methods of alternate coding (26%) or receive feedback on denied claims (46%). Most physicians believe that alternate coding is justified in the present system; however, some physicians expressed concerns that these practices may contribute to stigmatization or lead to improper management decisions. CONCLUSIONS: Alternate coding is commonly reported; however, approaches to diagnostic coding vary by provider specialty. Reimbursement issues are important, but other challenges in diagnosis and classification hold special relevance to children with behavioral and mental disorders. There seems to be a great need to reconsider the separate goals and uses of clinical diagnosis and administrative coding. Additional study is needed to assess how reported coding practices may affect administrative data, patient care, and health care economics. PMID- 12093990 TI - Intravenous ribavirin treatment for severe adenovirus disease in immunocompromised children. AB - BACKGROUND: Adenovirus is an important cause of morbidity and mortality in the immunocompromised host. The incidence of severe adenovirus disease in pediatrics is increasing in association with growing numbers of immunocompromised children, where case fatality rates as high as 50% to 80% have been reported. There are no approved antiviral agents with proven efficacy for the treatment of severe adenovirus disease, nor are there any prospective randomized, controlled trials of potentially useful anti-adenovirus therapies. Apparent clinical success in the treatment of severe adenovirus disease is limited to a few case reports and small series. Experience is greatest with intravenous ribavirin and cidofovir. Ribavirin, a guanosine analogue, has broad antiviral activity against both RNA and DNA viruses, including documented activity against adenovirus in vitro. Ribavirin is licensed in aerosol form for the treatment of respiratory syncytial virus infection, and orally in combination with interferon to treat hepatitis C. Intravenous ribavirin is the treatment of choice for infection with hemorrhagic fever viruses. The most common adverse effect of intravenous ribavirin is reversible mild anemia. The use of cidofovir in severe adenovirus infection has been limited by adverse effects, the most significant of which is nephrotoxicity. OBJECTIVE: We report our experience with intravenous ribavirin therapy for severe adenovirus disease in a series of immunocompromised children and review the literature. DESIGN/METHODS: We retrospectively reviewed the medical records of 5 children treated with intravenous ribavirin for documented severe adenovirus disease. Two patients developed adenovirus hemorrhagic cystitis after cardiac and bone marrow transplants, respectively. The bone marrow transplant patient also received intravenous cidofovir for progressive disseminated disease. An additional 3 children developed adenovirus pneumonia; 2 were neonates, 1 of whom had partial DiGeorge syndrome. The remaining infant had recently undergone a cardiac transplant. Intravenous ribavirin was administered on a compassionate-use protocol. RESULTS: Complete clinical recovery followed later by viral clearance was observed in 2 children: the cardiac transplant recipient with adenovirus hemorrhagic cystitis and the immunocompetent neonate with adenovirus pneumonia. The remaining 3 children died of adenovirus disease. Intravenous ribavirin therapy was well tolerated. Use of cidofovir in 1 child was associated with progressive renal failure and neutropenia. DISCUSSION: Our series of patients is representative of the spectrum of immunocompromised children at greatest risk for severe adenovirus disease, namely solid-organ and bone marrow transplant recipients, neonates, and children with immunodeficiency. Although intravenous ribavirin was not effective for all children with severe adenovirus disease in this series or in the literature, therapy is unlikely to be of benefit if begun late in the course of the infection. Early identification, eg by polymerase chain reaction of those patients at risk of disseminated adenovirus disease may permit earlier antiviral treatment and better evaluation of therapeutic response. CONCLUSIONS: Two of 5 children with severe adenovirus disease treated with intravenous ribavirin recovered. The availability of newer rapid diagnostic techniques, such as polymerase chain reaction, may make earlier, more effective treatment of adenovirus infection possible. Given the seriousness and increasing prevalence of adenovirus disease in certain hosts, especially children, a large, multicenter clinical trial of potentially useful anti-adenoviral therapies, such as intravenous ribavirin, is clearly required to demonstrate the most effective and least toxic therapy. PMID- 12093991 TI - Factors associated with permanent closure of the ductus arteriosus: a role for prolonged indomethacin therapy. AB - BACKGROUND: The most important factor determining anatomic remodeling and permanent closure of the ductus arteriosus is the degree of ductus constriction after indomethacin treatment. Muscular constriction produces a region of ischemic hypoxia in the middle of the ductus muscle media that initiates the process of permanent closure. Previous studies have shown that infants delivered before 28 weeks' gestation, who still have evidence of ductus flow on Doppler examination (performed after the standard 3-dose course of indomethacin), have a high likelihood (>85% chance) of reopening their ductus in the future. In contrast, if there is no evidence of luminal patency on the posttreatment Doppler examination, the incidence of ductus reopening is <20%. In the following study, we examined infants who still had a patent ductus on Doppler examination after a 3-dose course of indomethacin, to identify which factors might be associated with permanent ductus closure. We hypothesized that infants who received additional doses of indomethacin after the standard 3-dose course might develop an even tighter degree of ductus constriction and increase their chance of developing permanent closure. METHODS: We performed a retrospective cohort study of preterm infants (< or =26; weeks' gestation) who were treated with indomethacin. Between 12 and 24 hours after the third dose of indomethacin, infants were examined for the presence or absence of ductus-related signs, and an echocardiogram was performed. Infants responded to the initial 3 doses of indomethacin in 1 of 3 ways: 1) the ductus was closed clinically (absent clinical signs) with no evidence of luminal flow on Doppler examination ("clinically closed"; n = 214); 2) the ductus was closed clinically, but a small amount of left-to-right luminal flow was evident on Doppler examination ("partially closed"; n = 69); 3) or the ductus was open clinically and echocardiographically ("nonresponder"; n = 30). Nonresponders underwent surgical ligation (n = 30). Infants with a partially closed ductus formed our study population. We used a hierarchical regression model to identify which, if any, of the following factors might be associated with permanent anatomic closure in the 69 infants with a partially closed ductus: 1) gestational age, 2) exposure to antenatal steroids, 3) birth weight, 4) sex, 5) presence and severity of respiratory distress, 6) fluid administration during the first 96 hours after birth, 7) indomethacin treatment approach (prophylactic vs symptomatic), 8) year of birth, 9) use of surfactant, 10) preeclampsia, 11) chorioamnionitis, 12) bacterial septicemia, 13) necrotizing enterocolitis, or 14) duration of indomethacin treatment (standard 3-dose course vs prolonged 6-dose course). Infants who received the standard 3-dose course of indomethacin treatment were given 0.2, 0.1, and 0.1 mg/kg indomethacin during a 48-hour period. Infants who received the prolonged 6-dose course of indomethacin treatment were given a fourth, fifth, and sixth dose of 0.1 mg/kg at 24 hour intervals, starting 24 hours after the third dose. RESULTS: Sixty-eight of the 69 infants survived long enough to complete all of the study evaluations. Seventy five percent (51/68) reopened their ductus and became symptomatic; 71% (48/68) were eventually ligated. Only gestational age and duration of indomethacin treatment were significantly and independently associated with permanent closure. A prolonged 6-dose course of indomethacin was more likely than the standard 3 dose course to be associated with an increased incidence of echocardiographic closure, a decreased incidence of symptomatic reopening (odds ratio: 0.19; 95% confidence interval: 0.04-0.96), and a decreased incidence of ductus ligation (odds ratio: 0.14; 95% confidence interval: 0.03-0.68). DISCUSSION: Several older studies have suggested that a longer initial course of indomethacin therapy may be more effective in producing permanent ductus closure than the standard 3-dose course. In contrast, more recent studies have found that a longer course of indomethacin is no more effective than the standard 3-dose course in producing permanent closure. We hypothesize that the different outcomes among these studies may be attributable to differences in the degree of ductus constriction during the standard 3-dose course of indomethacin. Both the increased use of antenatal steroids and the earlier use of indomethacin has increased the effectiveness of the standard 3-dose course of indomethacin in recent years. We hypothesize that, in contrast with earlier studies, a significant proportion of the infants in the recent studies may have developed complete Doppler closure with just 3 doses of indomethacin (as occurred in 214 of the 313 infants treated with the standard 3 dose course in our study). Because the degree of ductus constriction seems to determine the rate of anatomic remodeling and permanent closure, daily echocardiographic evaluations of ductal patency may be the best way to decide when indomethacin therapy is no longer needed. Our study suggests that infants who still have evidence of luminal patency, after a standard 3-dose course of indomethacin, may be likely to benefit from a longer course of indomethacin. Future randomized trials that examine the benefits of different lengths of indomethacin treatment may wish to take this into consideration. CONCLUSIONS: Despite the increased effectiveness of a prolonged course of indomethacin, the rates of ductus reopening and surgical ligation were still very high in infants with a partially closed ductus. Other therapeutic approaches will need to be developed before permanent closure is likely to occur in this group of immature infants. PMID- 12093992 TI - Can child deaths be prevented? The Arizona Child Fatality Review Program experience. AB - OBJECTIVE: To determine the causes and preventability of child deaths; to assess the accuracy of death certificate information; and to assess the number of child abuse deaths that are misdiagnosed as deaths attributable to natural or accidental causes. METHODS: Analysis of deaths of children <18 years old that occurred between 1995-1999 using the data collected by the Arizona Child Fatality Review Program (ACFRP). RESULTS: From 1995-1999, local multidisciplinary child fatality review teams (CFRTs) have reviewed 95% of all deaths of children <18 years old in Arizona. Each team has access to the child's death certificate, autopsy report, hospital records, child protective services records, law enforcement reports, and any other relevant documents that provide insight into the cause and preventability of a child's death. After reviewing these documents, the team determines the cause of death, its preventability, and the accuracy of the death certificate. The ACFRP defines a child's death as preventable if an individual or the community could reasonably have done something that would have changed the circumstances that led to the child's death. The ACFRP determined that 29% (1416/4806) of these deaths could have been prevented, and preventability increased with the age of the child. Only 5% (81/1781) of neonatal deaths were considered preventable, whereas the deaths of 38% of all children older than 28 days were considered preventable. By 9 years of age, the majority of child deaths (56%) were considered preventable. Deaths attributable to medical conditions were far less likely to be considered preventable than deaths attributable to unintentional injuries. Although 62% of all deaths in Arizona during the 5-year period were attributable to medical conditions, only 8% (253/2983) of these deaths were considered preventable. In contrast, 91% (852/934) of the deaths attributable to unintentional injuries were considered preventable. Motor vehicle crashes accounted for 634 of the deaths resulting from injuries, and drowning accounted for 187 deaths. Motor vehicle crashes were the leading cause of death for all children in Arizona over 1 year of age. Only 18% of child passengers and 3% of adolescent drivers who died were known to be appropriately restrained. The typical drowning victim was a young child who drowned in the family's backyard pool. Indeed, 70% (131/187) of the drowning victims were <5 years old, and 62% (81/131) of these children died in a backyard pool. Supervision of the child and pool fencing could have prevented 90% of these deaths. Most deaths attributable to medical conditions occurred in the first year of life. Prematurity was the most common medical condition (1036 deaths) followed by congenital anomalies (662 deaths) and infectious diseases (470 deaths). Some of the reasons why CFRTs believed a medical death was preventable included inadequate emergency medical services, poor continuity of care, and delay in seeking care because of lack of health insurance. There were 4 deaths resulting from infections that were vaccine-preventable. There were 263 deaths attributable to sudden infant death syndrome. Only 38 of these infants were found lying on their back; 35 were found lying on their side. The death rate from sudden infant death syndrome decreased from 1.1 per 1000 infants <1 year of age in 1995 to 0.5 in 1999. There were 33 deaths that the CFRTs concluded were attributable to unsafe sleeping arrangements that resulted in unintentional suffocation. From 1995-1999, 317 Arizona children died from gun shot wounds. Most of these deaths were homicides (175) or suicides (109). All suicide deaths occurred in children >9 years old, and 77% of these children were >14 years old. The typical suicide victim was male (83%) and used a gun (70%) to kill himself. After review by the CFRTs, it was determined that 5 of the 67 child abuse deaths were misdiagnosed as attributable to natural or accidental causes on the death certificate. In 3 of these 5 cases, the child was in a persistent vegetative state and died many years after the episode of child abuse. Although inaction or inappropriate action by Child Protective Services (CPS) is often thought to be the cause of child abuse deaths, the ACFRP determined that in 79% of child abuse deaths, there had been no previous CPS involvement with the child's family. Although 61% of child abuse deaths were considered to be preventable, much of the responsibility for preventing these deaths rests with community members (eg, relatives, neighbors) who were aware of the abuse but failed to report the family to CPS. The CFRTs, who had received training in the proper completion of death certificates, reported that the cause of death was incorrect on 13% of all death certificates and in 16 cases, the CFRTs disagreed with the medical examiner on the manner of death (eg, natural, accidental, undetermined). Because CFRTs have access to additional information that may not have been available to the physician who completes a child's death certificate, CFRTs may be able to more accurately determine the cause and manner of death than the physician who completed the death certificate. CONCLUSIONS: Arizona's child death rate is above the national average (82.16/100 000), but the ACFRP determined that many of these deaths could have been prevented by using known prevention strategies (eg, child safety restraints, pool fencing). Most child mortality data are based on death certificate information that often is incorrect and cannot be used to assess preventability. Although most states have child fatality review programs that review suspected child abuse deaths, <3% of all preventable deaths in Arizona were attributable to child abuse. If all child deaths in the United States were reviewed from a prevention/needs assessment perspective, targeted and data-driven recommendations for prevention could be developed for each community, and potentially 38% of all child deaths that occur after the first month of life could be prevented. The ACFRP is an excellent example of a statewide system with a public health focus. To assist other states in developing similar programs, national support is needed. The establishment of a public health focused federally funded national program would provide us with the opportunity to standardize data collection among states and better utilize this data at a national level. PMID- 12093993 TI - Absolute band counts in febrile infants: know your laboratory. AB - It was the impression of 1 of the authors that band counts in febrile infants < or =60 days of age were much higher as reported from the clinical laboratory at Children's Hospital Medical Center of Akron, Ohio, than they had been at Strong Memorial Hospital in Rochester, New York. Absolute band counts (ABC) from 119 febrile infants < or =60 days of age seen in the emergency department of Children's Hospital Medical Center of Akron for whom blood culture results were known were obtained retrospectively and compared with results from a prospective study conducted in Rochester. In Akron, 45% of the infants had elevated band counts and 16% had no other risk factor for serious bacterial infection compared with 5.9% and 1.4%, respectively, in Rochester. The ABC can vary widely from laboratory to laboratory. The clinician must use caution when using the ABC as a criterion for identifying infants at low risk for serious bacterial infection. PMID- 12093994 TI - Familial arachnoid cysts in association with autosomal dominant polycystic kidney disease. AB - Autosomal dominant polycystic kidney disease (ADPKD) is a systemic disorder well known for its association with intracranial aneurysms. Recently, intracranial arachnoid cysts have also been reported to be associated with ADPKD. We describe a father and daughter who each has a posterior fossa arachnoid cyst and asymptomatic ADPKD. To our knowledge, this is the first report of familial occurrence of arachnoid cysts in association with ADPKD. PMID- 12093995 TI - Dysmetabolic syndrome: multiple risk factors for premature adult disease in an adolescent girl. AB - The clinical diagnosis of dysmetabolic syndrome in an adult defines a patient with abnormal glucose metabolism (or diabetes), hypertension, hyperlipidemia, and obesity. This disorder accelerates atherosclerosis and significantly raises the risk for cardiovascular events. With the marked rise in the prevalence of obesity in childhood, obesity-linked risk factors are being expressed at young ages. The case of a 12-year-old girl with dysmetabolic syndrome is described and discussed. Emerging clinical data now indicate that the presence of 1 risk factor for cardiovascular disease in an overweight child should prompt screening for additional clinical abnormalities, with the aim of finding treatable disorders. PMID- 12093996 TI - Management of child and adolescent obesity: study design and practitioner characteristics. AB - OBJECTIVE: A study was undertaken to examine the attitudes and practices of health care providers in the assessment and treatment of overweight and obese children and adolescents. This study describes the study design and the practice settings and person characteristics of the practitioners included in this study. METHODS: A needs assessment questionnaire was developed by a working group consisting of researchers, clinicians, educators, and representatives of the Maternal and Child Health Bureau, Health Resources and Services Administration (Department of Health and Human Services), National Center for Education in Maternal and Child Health, International Life Sciences Institute, and Harris Interactive, Inc. The questionnaire consisted of 35 questions divided into 3 topic areas and was disseminated to a sample of pediatricians (n = 1088), pediatric nurse practitioners (n = 879), and registered dietitians (n = 1652). RESULTS: Despite a low response rate (33% for pediatric nurse practitioners, 27% for registered dieticians, and 19% for pediatricians), descriptive data were obtained about a variety of practitioner characteristics. Some significant differences were observed across practitioner groups and between genders in regard to years in practice, body mass index, and dietary and physical activity behaviors. Significant relationships were also observed in some practitioner groups between body mass index and compliance with dietary and physical activity guidelines. CONCLUSIONS: Our data show there is a wide variance in practitioner characteristics, particularly in regard to gender, years of practice, body mass index, and obesity-related behaviors. It is hoped the analyses presented in this and in the subsequent articles will provide useful information on current attitudes and practices and will contribute to improvements in the treatment of overweight children and adolescents. PMID- 12093997 TI - Management of child and adolescent obesity: attitudes, barriers, skills, and training needs among health care professionals. AB - OBJECTIVE: The primary aim of this study was to evaluate among health care professionals their attitudes, perceived barriers, perceived skill level, and training needs in the management of child and adolescent obesity. METHODS: A national needs assessment consisting of a mailed questionnaire was conducted among a random sample of health care professionals. The survey was completed by 202 pediatricians, 293 pediatric nurse practitioners, and 444 registered dietitians. RESULTS: The majority of all respondents felt that childhood obesity was a condition that needs treatment (75%-93%), and affects chronic disease risk (76%-89%) and future quality of life (83%-93%). The most frequent barriers were lack of parent involvement, lack of patient motivation, and lack of support services. Registered dietitians were less likely to identify barriers to treatment compared with pediatricians or pediatric nurse practitioners. The most common areas of self-perceived low proficiency were in the use of behavioral management strategies, guidance in parenting techniques, and addressing family conflicts. All 3 groups expressed high interest in additional training on obesity management of children and adolescents, especially in the area of behavioral management strategies and parenting techniques. Those practitioners with >10 years of practice reported the greatest interest in training. CONCLUSIONS: Pediatric practitioners view child and adolescent obesity with concern and feel that intervention is important. However, several important barriers interfere with treatment efforts and will need to be addressed. There is also a need for increased training opportunities related to obesity prevention and treatment. The results of this study provide directions and priorities for training, education, and advocacy efforts. PMID- 12093998 TI - Management of child and adolescent obesity: psychological, emotional, and behavioral assessment. AB - OBJECTIVE: The primary objective of this study was to describe the usual psychological, emotional, and behavioral evaluations of overweight children and adolescents to target education of pediatric health care providers. A secondary aim was to determine whether certain provider characteristics were associated with recommended evaluation practices. METHODS: A total of 203 pediatricians, 293 pediatric nurse practitioners, and 444 registered dietitians completed questionnaires about their evaluation of overweight children and adolescents (response rates 19%, 33%, and 27%, respectively). Results were examined for adherence to published recommendations and for associations with certain provider characteristics. RESULTS: All 3 groups generally followed recommended psychological and emotional evaluation practices. Nearly all respondents obtained a diet history, and >75% evaluated physical activity as recommended. Fifty to 61% routinely initiated treatment in overweight children with no obesity-associated medical conditions, and 13% to 29% initiated treatment in children and adolescents who did not want to control weight. The majority included parents in treatment, but one third or fewer involved all household members. More than 75% sometimes referred to a mental health specialist. In general, provider's body mass index, gender, years in practice, and practice specialty were not associated with following evaluation recommendations. CONCLUSIONS: Emotional and behavioral assessments were generally consistent with recommendations. Providers will benefit from identification of methods to engage overweight youth who do not want to control weight. PMID- 12093999 TI - Medical evaluation of overweight children and adolescents: reports from pediatricians, pediatric nurse practitioners, and registered dietitians. AB - OBJECTIVE: The primary aim of this study was to determine how pediatric health care providers identify overweight in children and adolescents and how they evaluate obesity-related medical complications. This information can guide development of programs to help providers improve their evaluation practices. A secondary objective was to examine the association of certain provider characteristics with recommended evaluation practices. METHODS: A random sample of pediatricians, pediatric nurse practitioners (PNPs), and registered dietitians received a questionnaire about their evaluation of overweight children and adolescents. Results were compared with published recommendations. Associations between respondent characteristics and adherence to published recommendations were examined. RESULTS: A total 940 providers responded (response rate: 19%-33%). Among all 3 groups a majority frequently used clinical impression, weight-for-age percentile, weight-for-height percent, and weight-for-height percentile to assess degree of overweight. Nearly all pediatricians and PNPs routinely evaluated blood pressure, but a minority routinely looked for orthopedic problems, insulin resistance, and sleep disorders. Less than 10% followed all recommendations for history and physical examination. Two thirds of pediatricians and PNPs routinely tested for lipid abnormalities. Most providers asked about family history of overweight, hypertension, cardiovascular disease, and diabetes, but only one third asked about gallbladder disease. In general, the provider's specialty, years in practice, gender, and body mass index were not associated with adherence to recommended practices. CONCLUSIONS: Medical evaluation of overweight children and adolescents fell short of recommended practices. These results point to the need for educational efforts to increase awareness of medical risks and for tools to facilitate more complete evaluation during office visits. PMID- 12094000 TI - Treatment of child and adolescent obesity: reports from pediatricians, pediatric nurse practitioners, and registered dietitians. AB - OBJECTIVE: The primary aim of this study was to identify interventions used by pediatric health care providers in treatment of overweight children and adolescents to identify provider educational needs. A secondary aim was to examine the association of certain provider characteristics with recommended evaluation practices. STUDY DESIGN: A random sample of pediatricians, pediatric nurse practitioners, and registered dietitians (RDs) received questionnaires about their diet, activity, and medication recommendations for overweight patients and about referrals to specialists and programs. Results were examined for adherence to published recommendations and for associations with certain respondent characteristics. RESULTS: A total of 940 providers responded (response rate: 19%-33%). The majority recommended "changes in eating patterns" and "limitations of specific foods." Half or more used "low-fat diet" and "modest calorie restriction" in adolescents. Less than 15% used "very low-calorie diet." Fewer RDs recommended more restrictive diets. More than 60% of all groups followed recommended eating interventions for school-aged children and adolescents. More than 80% followed recommended physical activity interventions for all age groups. In each group, about 5% sometimes recommended prescription medication and herbal remedies for adolescents. None recommended surgery. Two thirds of pediatricians and pediatric nurse practitioners often referred to RDs. Approximately 20% referred to child/adolescent weight programs, but for 27% to 42%, these programs or pediatric obesity specialists were not available. No consistent associations between respondent characteristics and adherence to recommended interventions were identified. CONCLUSIONS: The providers generally promoted healthy eating and activity with minimal use of highly restrictive diets or medication to control weight. PMID- 12094001 TI - Management of child and adolescent obesity: summary and recommendations based on reports from pediatricians, pediatric nurse practitioners, and registered dietitians. PMID- 12094004 TI - Love in the time of Helicobacter. PMID- 12094005 TI - Might oral zinc protect pancreatic B-cells against oxidative insults? PMID- 12094006 TI - Evidence-based medicine and the need for non-commercial clinical research directed towards therapeutic innovation. PMID- 12094007 TI - Soluble T-lymphocyte antigen-specific immunoproteins: a progress report. AB - T-cell-derived proteins that bind nominal (non-MHC-associated) antigen specifically (TABM) express V and C region epitopes of the T-cell receptor (TCR) for antigen and have a significant similarity in amino acid sequence to TCR alpha chain V and C region. The presence of these immunoproteins in human serum and a specific increase in serum TABM in infectious disease, chemical sensitivity, and food intolerance suggest that TABM may impact on pathogenesis through the modulation of cell-mediated immunity, the antigen-specific concentration and delivery of immunoregulatory cytokines such as TGF-beta and elastase, and the induction of the release of substance P by sensory neurons. In this Minireview update, we describe advances in the detection and quantitation of human TABM by monoclonal antibodies, and the association of increased human serum TABM titers in infectious disease, chemical sensitivity, and food intolerance. We suggest that the immunomodulatory mode of action of these immunoproteins may be the antigen-specific focusing of cytokines associated with TABM. PMID- 12094008 TI - Pathophysiological effects of nicotine on the pancreas: an update. AB - Epidemiological evidence strongly suggests an association between cigarette smoking and pancreatic diseases. It is well recognized that nicotine, a major component in cigarette smoke, is an addictive agent and, therefore, reinforces smoking behavior. The current review update focuses on the genetics of nicotine dependence and its role on the development of pancreatic diseases. The role of smoking and nicotine in pancreatitis and pancreatic cancer development is also discussed. Exposure of laboratory animals to nicotine clearly supports the notion that nicotine can induce pancreatic injury. The mechanism by which nicotine induces such effects is perhaps mediated via signal transduction pathways in the pancreatic acinar cell, leading to enhanced levels of intracellular calcium release, resulting in cytotoxicity and eventual cell death. The induction of pancreatic injury by nicotine may also involve activation and expression of protooncogene, H-ras, which can increase cytosolic calcium via second messenger pathways. Development of pancreatic carcinoma in cigarette smokers as observed in human populations may be the result of activation and mutation of the H-ras gene. A possible pathogenetic mechanism of nicotine in the pancreas activating multiple signal transduction pathways is schematically summarized in Figure 1. PMID- 12094009 TI - Age-related changes in the regulation of luteinizing hormone secretion by estrogen in women. AB - Despite the many studies that have been conducted using both primate and human models to understand the control of the menstrual cycle, there are many aspects of the hormonal dynamics of the menstrual cycle that are not understood. This Minireview summarizes the changes in estrogen regulation of luteinizing hormone (LH) secretion that occur throughout life in women from the time of maturation of the hypothalamic-pituitary axis resulting in the occurrence of the LH surge during puberty, through the reproductive years, to the changes in the regulation of the LH surge during premenopause and, subsequently, menopause. PMID- 12094010 TI - The cyclooxygenase inhibitors indomethacin and Rofecoxib reduce regional cerebral blood flow evoked by somatosensory stimulation in rats. AB - The present study was designed to investigate whether administration of indomethacin (IMC), a non-selective cyclooxygenase (COX-1 and COX-2) inhibitor, and Rofecoxib, a highly selective COX-2 inhibitor, affect the regulation of regional cerebral blood flow response evoked by somatosensory activation (evoked rCBF). IMC and Rofecoxib were applied intravenously (6.25 and 3 mg/kg/hr, respectively). Somatosensory activation was induced by electrical hind paw stimuli of 0.2, 1, and 5 Hz (5-sec duration, 1.5 mA). The evoked rCBF was measured in alpha-chloralose anesthetized rats using laser-Doppler flowmetry. Before and after drug application, the evoked rCBF showed a frequency-dependent increase in the range of 0.2-5 Hz stimulation. IMC reduced significantly (about 50%-60%) evoked rCBF in response to all frequencies of hind paw stimulation (P< 0.05). Rofecoxib reduced significantly (about 50%) evoked rCBF in response to 1 and 5 Hz stimulation (P< 0.05), but did not affect evoked rCBF at 0.2 Hz. After IMC or Rofecoxib application, the normalized evoked rCBF curves peaked earlier as compared with that before their application (P< 0.05), although the rise time of 0.5 sec was nearly constant regardless of the stimulus frequency. The termination time of evoked rCBF curves was changed significantly after IMC application at 0.2 Hz stimulation (P< 0.05), but was not affected after Rofecoxib application. Neither COX inhibitor significantly affected the baseline level of CBF. The results suggest a participation of COX products in the regulation of evoked rCBF in response to somatosensory stimulation in the brain. PMID- 12094011 TI - Serial transplantation of p53-deficient hemopoietic progenitor cells to assess their infinite growth potential. AB - Thirty-five years ago, Siminovitch et al. (Siminovitch L, Till JE, McCulloch EA. J Cell Com Physiol 64:23-32, 1964), using serially transplanted mouse spleens at 14-day intervals, observed a markedly progressive decline in the proliferative capacity of bone marrow (BM) cells, with the loss of clonogenicity by the fourth transplant generation. Using the same protocol, we assessed the proliferative capacity of p53-deficient mouse BM cells transplanted serially at the same 14-day intervals into lethally irradiated mice, which was a useful tool for understanding the characteristics of hemopoietic stem cells lacking solely the p53 gene function. BM cells from p53-deficient homozygous (p53(-/-)), p53 heterozygous (p53(+/-)), and wild-type (p53(+/+)) C57BL/6 mice were transplanted into lethally irradiated C57BL/6 recipients. Fourteen days later, the repopulated spleens were harvested, and 10(7)cells were retransplanted into secondary recipients. Serial transplantation was continued at 14-day intervals until hemopoietic repopulation failure. The number of heterozygous and homozygous p53 deficient spleen cells increased logarithmically up to the fourth and fifth passages, respectively, whereas wild-type spleen cells ceased to proliferate by the third passage. The number of macroscopic spleen colonies increased logarithmically until the third passage in recipients of heterozygous and homozygous p53-deficient cells, but ceased to grow by the second passage in recipients of wild-type cells. The numbers of heterozygous and homozygous p53 deficient colony forming units in spleen (CFUs-S) remained stable during the first four transplant generations, whereas that of wild-type CFUs-S decreased progressively from the first transplant generation onward. The clonogenicity of p53-deficient cells was lost when the number of CFUs-S per spleen decreased to below 10. This suggests that one out of 10 CFUs-S might be long-term repopulating cells (LTRCs), and that p53-deficient LTRCs may proliferate more rapidly than wild-type LTRCs. Longer passages that were possible in the p53-deficient groups were considered to be due to the faster cell cycle of the p53-deficient hemopoietic progenitor cells, as determined by bromodeoxyuridine incorporation with purging by UV light exposure, followed by hemopoietic colony assay (BUUV assay). PMID- 12094012 TI - Characterization of acetylcholinesterase in Caco-2 cells. AB - Acetylcholinesterase (acetylcholine hydrolase, EC 3.1.1.7) was solubilized from cultured Caco-2 cells. It was established that this enzyme activity is acetylcholinesterase by substrate specificity (acetylthiocholine, acetyl-beta methylthiocholine>propionylthiocholine>butyrylthiocholine), substrate inhibition, and specificity of inhibitors (BW284c51>iso-OMPA). The acetylcholinesterase activity increased proportional to the degree of differentiation of the cells. Most of the enzyme was membrane bound, requiring detergent for solubilization, and the active site faced the external fluid. Only one peak of activity, which corresponded to a monomeric form, could be detected on linear sucrose density gradients. The sedimentation of this form of the enzyme was shifted depending on whether Triton X-100 or Brij 96 detergent was used. These results indicate that the epithelial-derived Caco-2 cells produce predominantly an amphiphilic, monomeric form of acetylcholinesterase that is bound to the plasma membrane and whose catalytic center faces the extracellular fluid. PMID- 12094013 TI - Neointima formation in the rat carotid artery is exacerbated by dietary copper deficiency. AB - Dietary copper is an essential trace element with roles in both functional and structural aspects of the cardiovascular system. In particular, the vascular response to inflammatory stimuli is known to be significantly augmented in copper deficient rats. The current study was designed to quantify the extent of injury induced neointimal proliferation and stenosis in rats fed diets either adequate or deficient in copper. Male, weanling Sprague-Dawley rats were fed purified diets that were either adequate (CuA; 5.6 microg Cu/g) or deficient (CuD; 0.3 microg Cu/g) in copper for 4 weeks. Balloon injury was induced in the left external carotid arteries. Fourteen days after injury, histomorphometric analysis of cross-sections from carotid arteries showed increased neointimal formation in the CuD group compared with the CuA controls (neointima/media ratio: 4.55 +/- 0.93 vs 1.45 +/- 0.2, respectively). These results correspond with data indicating that the activity of Cu/Zn-superoxide dismutase (SOD) is depressed in rats fed this CuD diet. Because superoxide anion and redox status are known to play a key role in the extent of neointimal formation in response to injury, we propose that the exaggerated neointimal proliferation seen in the CuD group is the result of the diminished Cu/Zn-SOD activity. PMID- 12094014 TI - Estrogen-dependent growth of a rat pituitary tumor involves, but does not require, a high level of vascular endothelial growth factor. AB - Long-term (10-week) treatment of Fischer 344 (F344) rats with the synthetic estrogen diethylstilbestrol (DES) increases the level of vascular endothelial growth factor (VEGF) in the pituitary. This is concurrent with the development of a large tumor of the pituitary of F344 rats. A role for VEGF in estrogen dependent pituitary tumor growth is also supported by the fact that pituitary VEGF level is not increased by estrogen treatment in rats of the tumor-resistant Brown Norway (BN) strain. However, VEGF is not increased by estrogen treatment in an F(1) hybrid of F344 and BN, even though F(1) hybrid rats do form pituitary tumors in response to estrogen. Quantitative trait locus (QLT) mapping reveals that control of estrogen-dependent VEGF expression is linked to the Edpm5 QTL, which was previously identified as a QTL for estrogen-dependent pituitary tumor growth. In contrast, the QTL Edpm2-1 and Edpm9-2, which have been shown to each have a significant effect on estrogen-dependent pituitary mass of a magnitude similar to Edpm5, do not have any effect on VEGF level. Taken together, our results support the association of VEGF expression with growth of the estrogen induced rat pituitary tumor, as has been reported by others, but they also indicate that there is significant pathways of growth regulation that are independent of high-level VEGF expression. PMID- 12094015 TI - Longer life spans and delayed maturation in wild-derived mice. AB - Nearly all the experimental mice used in aging research are derived from lineages that have been selected for many generations for adaptation to laboratory breeding conditions and are subsequently inbred. To see if inbreeding and laboratory adaptation might have altered the frequencies of genes that influence life span, we have developed three lines of mice (Idaho [Id], Pohnpei [Po], and Majuro [Ma]) from wild-trapped progenitors, and have compared them with a genetically heterogeneous mouse stock (DC) representative of the laboratory adapted gene pool. Mean life span of the Id stock exceeded that of the DC stock by 24% (P < 0.00002), and maximal life span, estimated as mean longevity of the longest-lived 10% of the mice, was also increased by 16% (P < 0.003). Mice of the Ma stock also had a significantly longer maximal longevity than DC mice (9%, P = 0.04). The longest-lived Id mouse died at the age of 1450 days, which appears to exceed the previous longevity record for fully fed, non-mutant mice. The life table of the Po mice resembled that of the DC controls. Ma and Id mice differ from DC mice in several respects: both are shorter and lighter, and females of both stocks, particularly Id, are much slower to reach sexual maturity. As young adults, Id mice have lower levels of insulin-like growth factor 1 (IGF-I), leptin, and glycosylated hemoglobin compared with DC controls, implicating several biochemical pathways as potential longevity mediators. The results support the idea that inadvertent selection for rapid maturation and large body size during the adaptation of the common stocks of laboratory mice may have forced the loss of natural alleles that retard the aging process. Genes present in the Id and Ma stocks may be valuable tools for the analysis of the physiology and biochemistry of aging in mice. PMID- 12094016 TI - Neuroprotective and neurotrophic efficacy of phytoestrogens in cultured hippocampal neurons. AB - Epidemiological data from retrospective and case-control studies have indicated that estrogen replacement therapy (ERT) can decrease the risk of developing Alzheimer's disease. In addition, ERT has been found to promote cellular correlates of memory and to promote neuronal survival both in vivo and in vitro. Phytoestrogens have been proposed as potential alternatives to ERT. To determine whether phytoestrogens exert estrogen agonist effect in neural tissue, investigations of neuroprotective and neurotrophic efficacy of phytoestrogens were conducted. Six phytoestrogens, genistein, genistin, daidzein, daidzin, formononetin, and equol, were tested for their neuroprotective efficacy against two toxic insults, glutamate excitotoxicity and beta-amyloid(25-35). Neuronal membrane damage was quantitatively measured by lactate dehydrogenase (LDH) release, and neuronal mitochondrial viability was determined by 3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromid (MTT) assay. Results of these studies demonstrated that all phytoestrogens induced a modest but significant reduction in LDH release following exposure to glutamate and beta amyloid(25-35). In contrast, none of phytoestrogens induced a significant increase in reduced MTT levels, which occurred in the presence of a full estrogen agonist, 17beta-estradiol. Analysis of the neurotrophic potential of genistein and daidzein, two phytoestrogens that exerted a significant reduction in LDH release, demonstrated that neither of these molecules promoted hippocampal neuron process outgrowth. Results of these analyses indicate that although phytoestrogens exert a neuroprotective effect at the plasma membrane, they do not sustain neuron mitochondrial viability nor do they induce cellular correlates of memory as neurite outgrowth and synaptogenesis are putative mechanisms of memory. Data derived from these investigations would predict that phytoestrogens could exert some neuroprotective effects analogous to that of antioxidants, but that these molecules are not functional equivalents to endogenously active 17beta estradiol or to estrogen replacement formulations and, therefore, would raise the concern that they may not reduce the risk of Alzheimer's disease or sustain memory function in postmenopausal women. PMID- 12094017 TI - Influence of ischemic preconditioning on intracellular sodium, pH, and cellular energy status in isolated perfused heart. AB - The possible relationships between intracellular Na(+) (Na(i)(+)), bioenergetic status and intracellular pH (pH(i)) in the mechanism for ischemic preconditioning were studied using (23)Na and (31)P magnetic resonance spectroscopy in isolated Langendorff perfused rat heart. The ischemic preconditioning (three 5-min ischemic episodes followed by two 5-min and one 10-min period of reperfusion) prior to prolonged ischemia (20 min stop-flow) resulted in a decrease in ischemic acidosis and faster and complete recovery of cardiac function (ventricular developed pressure and heart rate) after 30 min of reperfusion. The response of Na(i) during ischemia in the preconditioned hearts was characterized by an increase in Na(i)(+) at the end of preconditioning and an accelerated decrease during the first few minutes of reperfusion. During post-ischemic reperfusion, bioenergetic parameters (PCr/P(i) and betaATP/P(i) ratios) were partly recovered without any significant difference between control and preconditioned hearts. The reduced acidosis during prolonged ischemia and the accelerated decrease in Na(i)(+) during reperfusion in the preconditioned hearts suggest activation of Na(+)/H(+) exchanger and other ion transport systems during preconditioning, which may protect the heart from intracellular acidosis during prolonged ischemia, and result in better recovery of mechanical function (LVDP and heart rate) during post-ischemic reperfusion. PMID- 12094018 TI - Quantitative trait loci influencing hepatic copper in rats. AB - Significant differences in liver copper content have been observed between rat inbred strains. To define loci controlling this trait, the offspring (n = 190) from an (LEW/OlaHsd x BC/CpbU) F(2)-intercross was genetically analyzed. From each F(2) animal, liver copper content was determined and genomic DNA was screened with polymorphic DNA markers. We found a major quantitative trait locus (QTL) for liver copper content in females on chromosome 2 and in males on chromosome 10. Both QTLs accounted for approximately 20% of the genetic variance. In addition, suggestive linkage for liver copper content was found on rat chromosomes 1, 8, 10, 12, 14, and 19. The regions on these chromosomes contain genes that are responsible for 9.0-15.5% of the genetic variance of liver copper content. PMID- 12094019 TI - Humoral immune response in mice over-expressing or deficient in growth hormone. AB - Effects of growth hormone (GH) levels on the humoral immune response were investigated in metallothionein I (MT)-bovine (b) GH-transgenic (tg) and GH deficient Ames dwarf (Prop1 df(-/-)) mice. Four-month-old mice were given primary and secondary injections of either normal saline or tetanus toxoid (TT) to induce specific antibody (Ab) production. MT-bGH-tg mice with high peripheral levels of bGH produced less TT-specific Ab than normal nontransgenic (Ntg) littermates, df, or nondwarf (Ndf) control mice. Titers reached maximum levels at 3-4 weeks post primary immunization (PPI) and declined gradually through 24 weeks PPI in all groups of mice. Peripheral CD4(+) and CD8(+) T cell populations were significantly lower in tg than in Ntg, df, or Ndf mice. No significant differences were found in B cell numbers between tg, Ntg, or df mice. T helper 2 (Th2) cell populations were significantly greater in df mice compared to Ntg control mice. No significant differences were found in CD4(+):CD8(+) T cell ratios, interleukin (IL)-4 concentrations or interferon (IFN)-gamma levels between tg,Ntg, df, and Ndf mice. No patterns of significant sexual dimorphism were found for any of the immune parameters studied. Elevated levels of corticosterone were investigated as a possible immunosuppressant mechanism responsible for low Ab responses in the tg mice. Ab production was not enhanced by decreasing corticosterone in tg mice. Thus, high endogenous GH levels inhibit specific Ab production and peripheral T cell populations but not peripheral B cell numbers, Th2 cell populations, or IL-4 or IFN-gamma production. Elevated corticosterone levels do not appear to be responsible for suppressed humoral immune responses. Low levels of endogenous GH do not inhibit specific Ab production but may contribute to increased peripheral Th2 cell numbers. PMID- 12094022 TI - Sodium silver tricobalt bis(diphosphate) and sodium silver copper(II) diphosphate. AB - The crystal structures of two new diphosphates, sodium silver tricobalt bis(diphosphate), (Na(1.42)Ag(0.58))Co(3)(P(2)O(7))(2), and sodium silver copper(II) diphosphate, (Na(1.12)Ag(0.88))CuP(2)O(7), provide examples of the effect of mixing Na and Ag in the same site of known host phosphate compounds. The small differences in ionic radii of the two monocations do not lead to significant differences in the structural details. In the latter compound, the Cu atom lies on an inversion center. PMID- 12094020 TI - Absence of p53 and FasL has sexually dimorphic effects on both development and reproduction. AB - Reproduction and development are highly dependent on apoptosis to balance the proliferation that necessarily occurs during these processes. How the absence of two apoptotic factors in mice would affect reproduction and development was examined. Given previous reports of increased neural tube defects in p53-/- female fetuses, decreased fertility in gld female mice, and altered spermatogenesis in both p53 and gld male mice, the possibility that these phenotypes might be enhanced by the elimination of a second apoptotic factor was investigated. The reproductive vigor and the health of offspring were monitored during the production of the new double-deficient strain (FasL-/-p53-/-) for any changes from the reported phenotypes. Thus, any unusual phenotypes that could lead to new models for studying mechanisms of health and disease would be identified. Double-deficient male offspring appeared healthy and occurred at expected frequencies. Additionally, spermatogenesis and male fertility were unaffected by the gene deficiencies. On the other hand, FasL+/+p53-/- and FasL-/ p53-/- female mice were susceptible to increased malformations and post-natal death. These abnormalities were consistent with previous reports of neural tube defects in p53-/- female mice. Fertility rates were also significantly decreased in p53-/- female mice that lived to be adults, an observation not previously reported. Finally, the absence of both FasL and p53 led to dystocia in pregnant female mice, suggesting that the two genes play complementary roles in parturition. Therefore, although male mouse development and reproduction remained unaffected by p53 and FasL deficiencies, female mouse development was adversely affected by the absence of p53, and no live litters were born to female mice with the combined absence of both FasL and p53. In this report, we suggest a potential mechanism involving corpora luteal regression to explain this defect in parturition in FasL-/-p53-/- female mice. PMID- 12094021 TI - Differential development of aldehyde dehydrogenase in fore- and glandular stomach in postnatal rats. AB - Many foods contain the unsaturated aldehyde, hexadienal (HX). Human exposure is thus unavoidable. HX feeding to rodents caused cancers only in the forestomach. Aldehyde dehydrogenases (ALDH) are key enzymes in the metabolism of aldehydes. We examined the distribution of ALDH using HX as the substrate (HXDH) along the GI tract of adolescent rats and found that their stomachs have high levels of HXDH activity and the enzyme preferred HX > 9-cis-retinal > acetyl aldehyde > formyl aldehyde. We also followed the postnatal development of the stomach. At birth, the forestomach represented 40-50% of the total stomach weight. Both fore- and glandular stomach gained weight, with the glandular portion gaining at a faster rate. By 21 days, the forestomach was 24-28% of the total weight and decreased slightly to an adult level of 22-24%. Gastric HXDH is low from birth to 14 days of age. HXDH activity increased thereafter, reaching higher levels at 21 days and peaking around 30-36 days of age. The activity then decreased to the adult level. The fore- and glandular stomach had the same level of HXDH activity in the newborn and at 7 and 14 days of age. At weaning, HXDH activity was higher (3x) in the forestomach than in the glandular stomach. In adults, the forestomach still had 2x the HXDH activity compared to the glandular stomach. Zymograms showed similar isozyme patterns of HXDH but with different ratios of the three major forms between the forestomach and the glandular stomach. Results indicate a differential development of HXDH between the fore- and glandular stomach that might be related to the higher sensitivity of the forestomach to HX feeding. PMID- 12094023 TI - [K(2)CsYb(PO(4))(2)]. AB - The crystal structure of dipotassium caesium ytterbium bis(phosphate) is built up from regular independent PO(4) tetrahedra and YbO(6) octahedra sharing corners and arranged in layers. The structure is, in many respects, similar to that of glaserite. PMID- 12094024 TI - Trithallium hydrogen bis(sulfate), Tl(3)H(SO(4))(2), in the super-ionic phase by X-ray powder diffraction. AB - The structure of trithallium hydrogen bis(sulfate), Tl(3)H(SO(4))(2), in the super-ionic phase has been analyzed by Rietveld analysis of the X-ray powder diffraction pattern. Atomic parameters based on the isotypic Rb(3)H(SeO(4))(2) crystal in space group R3m in the super-ionic phase were used as the starting model, because it has been shown from the comparison of thermal and electric properties in Tl(3)H(SO(4))(2) and M(3)H(SO(4))(2) type crystals (M = Rb, Cs or NH(4)) that the room-temperature Tl(3)H(SO(4))(2) phase is isostructural with the high-temperature R3m-symmetry M(3)H(SO(4))(2) crystals. The structure was determined in the trigonal space group R3m and the Rietveld refinement shows that an hydrogen-bond O-H...O separation is slightly shortened compared with O-H...O separations in isotypic M(3)H(SeO(4))(2) crystals. In addition, it was found that the distortion of the SO(4) tetrahedra in Tl(3)H(SO(4))(2) is less than that in isotypic crystals. PMID- 12094025 TI - MgNB(9), a new magnesium nitridoboride. AB - The structure of a new magnesium nitridoboride, MgNB(9), has been refined from single-crystal X-ray data. The Mg and N atoms lie on sites with crystallographic 3m symmetry. The structure consists of two layers alternating along the c axis. The NB(6) layer, with B(12) icosahedra, has the C(2)B(13) structure type. Within this layer, boron icosahedra are bonded to N atoms, each coordinating to three boron polyhedra. Another MgB(3) layer, with B(6) octahedra, does not belong to any known structure type. The boron icosahedra and octahedra are connected to each other, thus forming a three-dimensional boron framework. PMID- 12094026 TI - Na(1.72)Mn(3.2)8(AsO(4))(3). AB - The single crystal of sodium manganese arsenate (1.72/3.28/12), Na(1.72)Mn(3.28)(AsO(4))(3), used for analysis was prepared by solid-state reaction at 1073 K. The compound crystallizes in the monoclinic system in space group C2/c. The structure consists of a complex network of edge-sharing MnO(6) octahedral chains, linked together by AsO(4) tetrahedra, forming two distinct channels, one containing Na(+) cations and the other occupied statistically by Mn(+) and Na(+) cations. PMID- 12094027 TI - Germanium(IV) chloride at 193 K. AB - Single crystals of germanium(IV) tetrachloride, GeCl(4), were grown at 193 K. In the crystal structure, the isolated molecules have very well realised tetrahedral geometry. PMID- 12094028 TI - Bis(trimethylphosphine)silver(I) hexafluorophosphate. AB - In the title two-coordinate silver compound, [Ag(C(3)H(9)P)(2)]PF(6), the cation has crystallographically imposed mirror symmetry, and approximates very closely to 3m (D(3d)) symmetry with fully staggered methyl groups in the solid state. The Ag atom has a nearly linear coordination geometry, with a P[bond]Ag[bond]P angle of 178.70 (4) degrees. The Ag[bond]P bond lengths are 2.3746 (12) and 2.3783 (12) A, which are significantly longer than the Au[bond]P bond length of 2.304 (1) A in the analogous two-coordinate gold cation. The lack of intramolecular steric effects within the present cations containing trimethylphosphine (cone angle 118 degrees), compared with those in known cations containing trimesitylphosphine (cone angle 212 degrees), provides a better comparison of M[bond]P distances and thus more conclusive evidence that Au really is smaller than Ag. PMID- 12094029 TI - (Dibenzyl sulfoxide-kappa O)bis(1,3-diphenylpropane-1,3-dionato kappa(2)O,O')dioxouranium(VI). AB - In the title compound, [UO(2)(C(15)H(11)O(2))(2)(C(14)H(14)OS)], the U(VI) atom is coordinated by seven O atoms in a distorted pentagonal-bipyramidal geometry. Both diphenylpropane-1,3-dionate systems are nearly planar. The sulfoxide moiety is in a distorted tetrahedral geometry, while its two aromatic rings are nearly orthogonal to one another. The crystal packing is stabilized by two bifurcated hydrogen-bonding interactions involving both uranyl O atoms. PMID- 12094030 TI - Disorder effects in Mn(12)-acetate at 83 K. AB - The structure of hexadeca-mu-acetato-tetraaquadodeca-mu(3)-oxo-dodecamanganese bis(acetic acid) tetrahydrate, [Mn(12)O(12)(CH(3)COO)(16)(H(2)O)(4)] x 2CH(3)COOH x 4H(2)O, known as Mn(12)-acetate, has been determined at 83 (2) K by X-ray diffraction methods. The fourfold (S(4)) molecular symmetry is disrupted by a strong hydrogen-bonding interaction with the disordered acetic acid molecule of solvation, which displaces one of the acetate ligands in the cluster. Up to six Mn(12) isomers are potentially present in the crystal lattice, which differ in the number and arrangement of hydrogen-bonded acetic acid molecules. These results considerably improve the structural information available on this molecular nanomagnet, which was first synthesized and characterized by Lis [Acta Cryst. (1980), B36, 2042-2046]. PMID- 12094031 TI - Chlorobis(thiourea-kappa S)copper(I) bis(4,5-diazafluoren-9-one) monohydrate. AB - The title compound, [CuCl(CH(4)N(2)S)(2)] x 2C(11)H(6)N(2)O x H(2)O, consists of molecules of a Cu(I)-thiourea complex, free 4,5-diazafluoren-9-one (dafone) and crystalline water. The planar complex molecule has trigonal coordination geometry around the Cu(I) atom. The dafone and water molecules, which are hydrogen bonded to the Cu(I) complex, are approximately coplanar with this complex. The crystal displays a sheet structure and pi-pi stacking is observed between neighbouring sheets. PMID- 12094032 TI - Di-mu-methoxy-bis[(eta(5)-cyclopentadienyl)(nitrosyl-kappa N)(trimethylsilylmethyl)molybdenum(II)]. AB - The title complex, [Mo(2)(C(5)H(5))(2)(CH(3)O)(2)(C(4)H(11)Si)(2)(NO)(2)], is formed in high yield by treating [CpMo(NO)(CH(2)SiMe(3))(2)] (Cp is cyclopentadienyl) with methanol. The nitrosyl ligands are nearly linear [O[bond]N[bond]Mo 170.1 (4) and 170.1 (5) degrees ], with short Mo[bond]N bonds [1.769 (4) and 1.776 (4) A] and long N[bond]O bonds [1.216 (5) and 1.201 (4) A]. The central four-membered Mo(2)O(2) ring exhibits an average Mo[bond]O bond length of 2.15 A. PMID- 12094033 TI - GaFPO(3)(C(6)H(5)): a new fluorinated gallium phenylphosphonate with a layered network. AB - Crystals of gallium fluorophenylphosphonate were synthesized hydrothermally at 453 K under autogenous pressure. The solid crystallizes in the monoclinic system and its structure is built up by the connection of zigzag chains of edge-sharing GaO(4)F(2) octahedra to phenylphosphonate groups. This results in the formation of a layered structure, in which the phenyl groups point upward and downward from the inorganic sheet. The Ga atoms occupy the special positions 4a (inversion center) and 4e (twofold axis). PMID- 12094034 TI - u-Cyano-1:2 kappa(2)C:N-tetracyano-1 kappa(4)C-(5-methyl-5-nitro-3,7-diazanonane 1,9-diamine-2 kappa(4)N(1,3,6,9))-nitrosyl-1 kappa N-copper(II)iron(II) dihydrate. AB - The title binuclear complex, [CuFe(CN)(5)(C(8)H(21)N(5)O(2))(NO)] x 2H(2)O or [CuFe(nelin)(CN)(5)(NO)] x 2H(2)O (nelin is 5-methyl-5-nitro-3,7-diazanonane-1,9 diamine) consists of discrete binuclear mixed-metal species, with a Cu centre linked to an Fe centre through a cyano bridge, and two water molecules of crystallization. In the complex, the Cu(II) ion is coordinated by five N atoms and has a distorted square-pyramidal geometry. The Fe(II) centre is in a distorted octahedral environment. PMID- 12094035 TI - [Bis(2-diphenylphosphinoethyl) sulfide-kappa(3)P,S,P'](triphenylphosphine-kappa P)platinum(II) diperchlorate acetone solvate. AB - In the title compound, [Pt(C(18)H(15)P)(C(28)H(28)P(2)S)](ClO(4))(2*-)C(3)H(6)O or [Pt(PPh(3))(PSP)](ClO(4))(2*) x CH(3)COCH(3), where PSP is the potentially tridentate chelate ligand bis(2-diphenylphosphinoethyl) sulfide, all three donor groups of the PSP ligand are coordinated to the central Pt atom, with Pt[bond]P = 2.310 (1) A and Pt[bond]S = 2.343 (1) A. The fourth coordination site is occupied by the P donor of the triphenylphosphine ligand [Pt[bond]P = 2.289 (1) A]. The complex cation has exact mirror symmetry, with the S atom, the Pt atom and the P atom of the PPh(3) ligand in the mirror plane. The Pt atom has a distorted square planar coordination geometry. A pi[bond]pi interaction is present between the phenyl rings of the PPh(3) ligand and the terminal -PPh(2) group of the PSP chelate. PMID- 12094036 TI - trans-Bis[O-2,4-di-tert-butylphenyl (4-methoxyphenyl)dithiophosphonato kappa(2)S,S']nickel(II). AB - In the the title compound, [Ni(C(21)H(28)O(2)PS(2))(2)], the Ni atom resides on an inversion centre and is coordinated in a square-planar array by four S atoms, with Ni[bond]S and P[bond]S bond lengths of 2.2336 (12)/2.2351 (13) and 1.9910 (16)/2.0010 (17) A, respectively. The two O-2,4-di-tert-butylphenyl and two 4 methoxyphenyl moieties adopt trans configurations about the central Ni atom. PMID- 12094037 TI - Tris[2-(2-oxazolin-2-yl)phenolato]iron(III). AB - The title compound, tris[2-(4,5-dihydrooxazol-2-yl-kappa N)phenolato-kappa O]iron(III), [Fe(C(9)H(8)NO(2))(3)], is disordered over a non-crystallographic twofold rotation axis perpendicular to the crystallographic threefold rotation axis. The disorder can be a pure rotational disorder of an iron complex in the facial configuration, or the consequence of a mixture of facial and meridional configurations. In the latter case, at least 25% of the iron complexes must adopt the facial configuration in order to obtain the disorder ratio observed in the crystal. PMID- 12094038 TI - A pyrazine bis-adduct of a binuclear rhodium(II) carboxylate containing 3,4,5 triethoxybenzoate as the equatorial ligand. AB - The title compound, tetrakis(mu-3,4,5-triethoxybenzoato kappa(2)O:O')bis[(pyrazine-kappa N)rhodium(II)](Rh[bond]Rh), [Rh(2)(C(13)H(17)O(5))(4)(C(4)H(4)N(2))(2)], crystallizes on an inversion centre in the triclinic space group P1. The equatorial carboxylate ligands bridge the two Rh(II) atoms, giving a binuclear lantern-like structure. The pyrazine molecules occupy the two axial coordination sites. The phenyl rings are tilted by ca 10 degrees with respect to the attached carboxylate groups. The pyrazine planes have a torsion angle of ca 19 degrees around the Rh-N bond with respect to the plane of the nearer carboxylate group and are not coplanar with the Rh[bond]Rh bond. PMID- 12094039 TI - catena-Poly[[[bis(mu-4,5-dihydrothiazole-2-thiolato)-tetrahedro-tetracopper(I)] di-mu-4,5-dihydrothiazole-2-thiolato] dihydrate]. AB - The title compound, [[Cu(4)(C(3)H(4)NS(2))(4)] x 2H(2)O](n), was produced by diffusing a solution of 2-mercaptothiazoline in tetrahydrofuran into a solution of CuCl in CH(3)CN at room temperature. The structure is characterized by self assembled one-dimensional chains that are condensed from butterfly-like [Cu(C(3)H(4)NS(2))](4) tetrameric units via double S-bridging at opposite ends. The Cu[bond]Cu distances within the Cu(4) butterfly cluster are in the range 2.7103 (10)-2.9764 (10) A, while the shortest Cu...Cu intercluster distance is 3.468 (1) A, much longer than the sum of the van der Waals radii. PMID- 12094040 TI - Aquachlorobis[4,4,5,5-tetramethyl-2-(2-pyridyl)-4,5-dihydro-1H-imidazol-1-oxyl 3 oxide-kappa(2)O(1),N(2)]nickel(II) nitrate methanol disolvate monohydrate. AB - The title complex, [NiCl(C(12)H(16)N(3)O(2))(2)(H(2)O)]NO(3) x 2CH(4)O x -H(2)O, was obtained from a methanolic solution of Ni(NO(3))(2) x 6H(2)O, 2-pyridyl nitronyl nitroxide (2-NITpy) and (NEt(4))(2)[CoCl(4)]. The equatorial coordination sites of the octahedral Ni(II) centre are occupied by two chelating radical ligands, with the axial positions occupied by the Cl(-) and water ligands. The H(2)O[bond]Ni[bond]Cl axis of the complex lies along a crystallographic twofold axis, so that only half the cation is present in the asymmetric unit. The Ni[bond]Cl bond length [2.3614 (17) A] is significantly shorter than distances typical of octahedral Ni(II) centres [2.441 (5) A]. However, with only one nitrate anion per formula unit, the oxidation state of the metal must be assigned as Ni(II). The 2-NITpy ligands bend away from the equatorial plane, forming a hydrophobic region around the Cl atoms. Conversely, the ligated water molecule forms moderately strong hydrogen bonds with the disordered methanol solvent molecules, which in turn form interactions with the water of crystallization and the disordered nitrate anion. These interactions combine to give hydrophilic regions throughout the crystal structure. PMID- 12094041 TI - Tetraaqua(1,10-phenanthroline-kappa(2)N,N')manganese(II) sulfate dihydrate. AB - The title compound, [Mn(C(12)H(8)N(2))(H(2)O)(4)]SO(4).2H(2)O, was obtained unexpectedly as a by-product from the reaction of sodium maleate, 1,10 phenanthroline (phen) and manganese sulfate tetrahydrate. The Mn atom is coordinated by the two N atoms of the phen ligand and four water O atoms in a highly distorted octahedral geometry, with Mn[bond]O distances in the range 2.155 (2)-2.203 (2) A and Mn[bond]N distances of 2.254 (2) and 2.272 (3) A. Extensive hydrogen-bonding interactions involving the water molecules and sulfate anions, and stacking interactions involving the phen rings, are observed in the crystal structure. PMID- 12094042 TI - Bis(tetra-n-butylammonium) di-mu-chloro-bis[(tetramethyl buta-1,3-diene-1,2,3,4 tetracarboxylato-kappa(2)C(1),C(4))palladium]. AB - The anionic complex in the title compound, (C(16)H(36)N)(2)[Pd(2)(C(12)H(12)O(8))(2)Cl(2)], lies on a centre of inversion, so that the [Pd(2)(mu-Cl)(2)] core is planar, which is the most frequent conformation found for complexes containing this moiety in the Cambridge Structural Database [October 2001 Release; Allen & Kennard (1993). Chem. Des. Autom. News, 8, 1, 31-37]. This dinuclear complex has a Pd...Pd distance of 3.5119 (4) A. The bite angle of the chelating ligand [79.79 (8) degrees] distorts the square-planar coordination around the metal atom. PMID- 12094043 TI - Dabconium orthofluoroberyllate hemihydrate. AB - Alternating C(6)H(14)N(2)(2+) and [BeF(4)](2-) units within the ac plane of 1,4 diazoniabicyclo[2.2.2]octane tetrafluoroberyllium hemihydrate, (C(6)H(14)N(2))[BeF(4)] x 0.5H(2)O, combine to form double chains via hydrogen bonding. These double chains connect through bifurcated hydrogen bonds to water molecules, which lie on twofold axes, producing a two-dimensional sheet. Very weak hydrogen-bond interactions, along the b axis, join the sheets to produce a three-dimensional network. PMID- 12094044 TI - Ethylenediammonium bis[bis[N-(2-aminoethyl)glycinato kappa(3)N,N',O]chromium(III)] tetrachloride dihydrate at 293 and 100 K. AB - The crystal structure of the title compound, (C(2)H(10)N(2))[Cr(C(4)H(9)N(2)O(2))(2)](2)Cl(4) x 2H(2)O, has been determined by single-crystal X-ray diffraction studies at 293 and 100 K. The analyses demonstrated that the crystal consists of ethylenediammonium dications (which lie about inversion centres), bis[N-(2-aminoethyl)glycinato]chromium(III) monocations, Cl(-) anions and hydrate water molecules, in a molecular ratio of 1:2:4:2. The complex cation unit has a slightly distorted octahedrally coordinated Cr atom, with two Cr[bond]O and four Cr[bond]N bonds in the ranges 1.951 (1)-1.953 (1) and 2.054 (1)-2.089 (2) A, respectively, at 293 K. The geometry of the bis[N-(2-aminoethyl)glycinato]chromium(III) moiety was found to be trans,cis,cis with respect to the carboxylate O atom and the primary and secondary amine N atoms. The two analyses, at 293 and 100 K, exhibited no remarkable structural differences, although the colour of the crystals did differ, being red at 293 K and orange at 100 K. PMID- 12094045 TI - (4-Nitrophenylsulfanylmethyl)triphenylstannane and (4 nitrophenylsulfonylmethyl)triphenylstannane: R(2)(2)(X) rings (X is 10, 18 or 24) and C[bond]H...pi interactions. AB - In the crystal structures of (4-nitrophenylsulfanylmethyl)triphenylstannane, [Sn(C(6)H(5))(3)(C(7)H(6)NO(2)S)], (I), and (4 nitrophenylsulfonylmethyl)triphenylstannane, [Sn(C(6)H(5))(3)(C(7)H(6)NO(4)S)], (II), the molecules are linked by paired C[bond]H...O hydrogen bonds into centrosymmetric dimers which combine to form sheets. In (I), two such dimers form to give R(2)(2)(10) and R(2)(2)(24) rings. In (II), similar dimers form, here with R(2)(2)(10) and R(2)(2)(18) rings, but with an additional dimer due to the presence of the sulfone group, giving R(2)(2)(10) rings. In both structures, C[bond]H...pi interactions lead to a doubling of the width of the sheets. PMID- 12094047 TI - Diaquabis(1,3-propanediamine-kappa(2)N,N')nickel(II) bis(sulfanilate). AB - The title compound, [Ni(C(3)H(10)N(2))(2)(H(2)O)(2)](C(6)H(6)NO(3)S)(2), contains alternating layers of sulfanilate anions and diaquabis(1,3 propanediamine)nickel(II) cations. The Ni atom lies on an inversion centre and is hexacoordinated by the 1,3-propanediamine ligands, which function as N,N' bidentate ligands, and the water molecules, which are in a trans arrangement. The sulfanilate anions are arranged in layers, with the sulfonate and amine groups directed towards opposite sides of the layer. The structure is stabilized by a network of hydrogen bonding between the O and N atoms of the sulfanilate anions, the water molecules, and the N atoms of the 1,3-propanediamine ligands. PMID- 12094046 TI - Losartan potassium, a non-peptide agent for the treatment of arterial hypertension. AB - In the title compound, potassium 2-butyl-4-chloro-1-[[2'-(5-tetrazolido)biphenyl 4-yl]methyl]-1H-imidazol-5-ylmethanol, K(+) x C(22)H(22)ClN(6)O(-), the imidazole and tetrazole rings are at angles of 85.0 (2) and 51.8 (1) degrees, respectively, to the phenyl rings to which they are attached, while the dihedral angle between the latter two rings is 46.7 (1) degrees. The coordination sphere of the metal cation consists of six tetrazoyl N atoms, the methanol O atom and the pi cloud of one of the phenyl rings. These interactions determine the formation of columns of molecular anions that lie parallel to the b axis, while hydrogen bonding contributes to intercolumnar cohesion. Far from the centre of the columns, the hydrocarbon chain is immersed in a hydrophobic environment. PMID- 12094048 TI - [2,6-Bis(3,5-dimethylpyrazol-1-ylmethyl)pyridine]iodocopper(I) dichloromethane solvate. AB - The title compound, [CuI(C(17)H(21)N(5))] x CH(2)Cl(2), contains a tetracoordinate Cu(I) centre with an unusual distorted tetrahedral stereochemistry, which has also been observed in other Cu(I) complexes containing this tridentate ligand. This distortion is probably a result of intermolecular steric contacts between the I(-) ligand and a neighbouring CH(2)Cl(2) molecule. PMID- 12094049 TI - 4,7-Diiodo-2,1,3-benzothiadiazole and 7,7'-diiodo-4,4'-bi(2,1,3 benzothiadiazole). AB - In the crystal structures of the title compounds, C(6)H(2)I(2)N(2)S, (I), and C(12)H(4)I(2)N(4)S(2), (II), respectively, a large number of short inter heteroatom contacts, such as S...N, I...I and N...I, are observed. In (II), which is non-centrosymmetric, two halves of the molecule are related by a crystallographic twofold axis. PMID- 12094050 TI - 5,11,17,23-Tetra-tert-butyl-25,26,27,28-tetrakis(2-cyanobenzyloxy)-2,8,14,20 tetrathiacalix[4]arene-dichloromethane (1/2). AB - A new p-tert-butylthiacalix[4]arene derivative, C(72)H(68)N(4)O(4)S(4) x 2CH(2)Cl(2), has been synthesized and is comprised of one tetra-p-tert butyltetrakis(2-cyanobenzyloxy)tetrathiacalix[4]arene and two dichloromethane molecules. The calix[4]arene molecule is centrosymmetric and adopts an unusual 1,2-alternate conformation via pi-pi interactions between adjacent cyanophenyl rings on the lower rim of the parent thiacalix[4]arene system. PMID- 12094051 TI - 4-Vinylbenzyl analogs of adenine and uracil: reactive monomers for nucleobase polymeric resins. AB - The crystal structures of 9-(4-vinylbenzyl)adenine, C(14)H(13)N(5), and 1-(4 vinylbenzyl)uracil, C(13)H(12)N(2)O(2), are composed of zigzag ribbon-like structures that are stabilized by conventional (N[bond]H...N-type) hydrogen bonds for the former and conventional (N[bond]H...O-type) and non-conventional (C[bond]H...O-type) hydrogen bonds for the latter; the hydrogen-bonding patterns are represented by graph-sets R(2)(2)(9) and R(2)(2)(8), respectively. The adenine and uracil moieties in these alkylated derivatives are planar and are inclined at angles of 84.44 (4) and 79.07 (7) degrees, respectively, with respect to the phenyl rings. PMID- 12094052 TI - Bis(5-phenyltetrazol-2-yl)methane. AB - In the title compound, alternatively named 5,5'-diphenyl-2,2' methyleneditetrazole, C(15)H(12)N(8), the dihedral angles between the tetrazole and benzene rings in the two 5-phenyltetrazole fragments are 2.45 (6) and 10.01 (9) degrees. There is weak intermolecular C[bond]H...N hydrogen bonding involving the H atoms of the methylene groups, which is responsible for the formation of two-membered aggregates. C[bond]H...pi interactions in the crystal structure are discussed. PMID- 12094053 TI - 2,4,6-Trimethylphenyl isocyanide. AB - The title compound, C(10)H(11)N, displays a crystallographic mirror plane that incorporates all the non-H atoms, as well as the H atoms attached to the aromatic ring. The isocyano group is almost linear and shows an N[triple bond]C bond distance of 1.158 (3) A. PMID- 12094054 TI - 1-Substituted derivatives of 2-phenylpyrrolidine-2-carboxamide. AB - The structures of two potential anti-human immunodeficiency virus type 1 (HIV-1) non-nucleoside reverse transcriptase inhibitors (NNRTI), namely 1-(2 chlorobenzoyl)-2-phenylpyrrolidine-2-carboxamide, C(18)H(17)ClN(2)O(2), and 1-(2 furoyl)-2-phenylpyrrolidine-2-carboxamide, C(16)H(16)N(2)O(3), have been investigated by X-ray diffraction and the butterfly-like conformation established in each case. The pyrrolidine ring has the same half-chair conformation in both structures. PMID- 12094056 TI - A pipecolic acid (Pip)-containing dipeptide, Boc-D-Ala-L-Pip-NH(i)Pr. AB - The title dipeptide, 1-(tert-butoxycarbonyl-D-alanyl)-N-isopropyl-L-pipecolamide or Boc-D-Ala-L-Pip-NH(i)Pr (H-Pip-OH is pipecolic acid or piperidine-2-carboxylic acid), C(17)H(31)N(3)O(4), with a D-L heterochiral sequence, adopts a type II' beta-turn conformation, with all-trans amide functions, where the C-terminal amide NH group interacts with the Boc carbonyl O atom to form a classical i+3 --> i intramolecular hydrogen bond. The C(alpha) substituent takes an axial position [H(alpha) (Pip) equatorial] and the trans pipecolamide function is nearly planar. PMID- 12094055 TI - Structural properties of a series of photochromic fluorinated indolylfulgides. AB - Fluorinated indolylfulgides are a class of photochromic organic compounds that meet many of the requirements for use as optical memory media and optical switches. The X-ray crystal structures of a series of five photochromic fluorinated indolylfulgides have been determined, namely (3Z)-3-[1-(1,2-dimethyl 1H-indol-3-yl)-2,2,2-trifluoroethylidene]-4-(1-methylethylidene)dihydrofuran-2,5 dione (trifluoromethylisopropylideneindolylfulgide), C(19)H(16)F(3)NO(3), (I), (3Z)-3-[1-(1,2-dimethyl-1H-indol-3-yl)-2,2,3,3,3-pentafluoropropylidene]-4-(1 methylethylidene)dihydrofuran-2,5-dione (pentafluoroethylisopropylideneindolylfulgide), C(20)H(16)F(5)NO(3), (II), (3Z)-3 [1-(1,2-dimethyl-1H-indol-3-yl)-2,2,3,3,4,4,4-heptafluorobutylidene]-4-(1 methylethylidene)dihydrofuran-2,5-dione (heptafluoropropylisopropylideneindolylfulgide), C(21)H(16)F(7)NO(3), (III), (3Z) 3-[1-(1,2-dimethyl-1H-indol-3-yl)-2,2,2-trifluoroethylidene]-4 (tricyclo[3.3.1.1(3,7)]decylidene)dihydrofuran-2,5-dione (trifluoromethyladamantylideneindolylfulgide), C(26)H(24)F(3)NO(3), (IV), and (3Z)-3-[1-(1,2-dimethyl-1H-indol-3-yl)-2,2,3,3,4,4,4-heptafluorobutylidene]-4 (tricyclo[3.3.1.1(3,7)]decylidene)dihydrofuran-2,5-dione (heptafluoropropyladamantylideneindolylfulgide), C(28)H(24)F(7)NO(3), (V). The photochromic property of fulgides is based on the photochemically allowed electrocyclic ring closure of a hexatriene system to form a cyclohexadiene. For each fulgide examined, the bond lengths within the hexatriene system alternate between short and long, as expected. Comparing the structures of the five fulgides with each other demonstrates no significant difference in bond lengths, bond angles or dihedral angles within the hexatriene systems. The distance between the bond-forming C atoms at each end of the hexatriene system does vary. Correlations of structural properties with optical properties are addressed. PMID- 12094057 TI - Bis(1-naphthyl) ditelluride. AB - The title compound, C(20)H(14)Te(2), shows a transoid conformation, with a C[bond]T[bond]-[bond]Te[bond]C torsion angle of 97.96 (9) degrees. The Te[bond]Te units show approximate eta(6) interactions with neighbouring naphthyl groups, forming chains along the c axis. The molecule lies about a crystallographic twofold axis. PMID- 12094058 TI - 1-Substituted derivatives of 2-aryl-5-oxopyrrolidine-2-carboxylic acid. AB - The structures of two potential anti-human immunodeficiency virus type 1 (HIV-1) non-nucleoside reverse transcriptase inhibitors (NNRTI), namely 1-benzyl-5-oxo-2 phenylpyrrolidine-2-carboxamide, C(18)H(18)N(2)O(2), (III), and 2-(4 isopropoxyphenyl)-5-oxo-1-(4-tolyl)pyrrolidine-2-carbonitrile, C(21)H(22)N(2)O(2), (IV), have been investigated by X-ray diffraction, confirming the butterfly-like conformation of both compounds. The pyrrolidine ring is in an envelope conformation in (III) and a half-chair conformation in (IV). Two intermolecular N[bond]H...O hydrogen bonds are present in the crystal structure of (III), with N...O distances of 2.995 (2) and 2.927 (2) A. PMID- 12094059 TI - (+)-3,11-Dioxoandrost-4-ene-17 beta-carboxylic acid: catemeric hydrogen bonding in a steroidal keto acid. AB - The title keto acid, C(20)H(26)O(4), forms carboxyl-to-ketone hydrogen-bonding catemers [O...O = 2.653 (5) A and O[bond]H...O = 172 (5) degrees ], linking translationally related molecules via the A-ring ketone. The two molecules in the cell form two parallel counter-directional chains, screw-related in b. A total of four intermolecular C[bond]H...O[double bond]C close contacts was found, involving both ketone functions. PMID- 12094060 TI - A benzopyran derivative substituted at position 3. AB - In (E)-3-[[(diphenoxyphosphoryl)methylhydrazono]methyl]-4H-1-benzopyran-4-one, C(23)H(19)N(2)O(5)P, the benzopyran-methylhydrazone moiety is planar and the two phenoxy phenyl rings are inclined at angles of 21.29 (6) and 89.33 (5) degrees. Weak C[bond]H...O and C[bond]H...N intramolecular interactions exert some influence on the planarity and orientation of that moiety. PMID- 12094061 TI - N-(2-hydroxyethyl)ethylenediammonium hydrogenphosphate monohydrate. AB - The title compound, C(4)H(14)N(2)O(2+) x HPO(4)(2-) x H(2)O, contains alternating interleaved layers of hydrogenphosphate and N-(2-hydroxyethyl)ethylenediammonium moieties. The water molecules are associated with channel-like voids in the structure and a network of hydrogen bonds stabilizes the crystal packing. PMID- 12094062 TI - Tetrameric triphenylsilanol, (Ph(3)SiOH)(4), and the adduct (Ph(3)SiOH)(2) dimethyl sulfoxide, both at 120 K, and the adduct (Ph(3)SiOH)(4)-1,4-dioxan at 150 K: interplay of O[bond]H...O and C[bond]H...pi(arene) interactions. AB - The structure of tetrameric triphenylsilanol, C(18)H(16)OSi, (I), has been re investigated at 120 (2) K. The hydroxyl H atoms were readily located and one of the arene rings is disordered over two closely positioned sets of sites. The molecules are linked into cyclic tetramers, having approximate 4 (S(4)) symmetry, via O[bond]H...O hydrogen bonds [H...O 1.81-1.85 A, O...O 2.634 (3)-2.693 (3) A and O[bond]H...O 156-166 degrees ]. At ambient temperature, there are indications of multiple disorder of the phenyl-ring sites. In bis(triphenylsilanol) dimethyl sulfoxide solvate, 2C(18)H(16)OSi x C(2)H(6)OS, (II), the dimethyl sulfoxide component is disordered across a twofold rotation axis in C2/c, and the molecular components are linked by a single O[bond]H...O hydrogen bond [H...O 1.85 A, O...O 2.732 (2) A and O[bond]H...O 172 degrees ] into three-molecule aggregates, which are themselves linked into a single three-dimensional framework by two C[bond]H...pi(arene) interactions. In tetrakis(triphenylsilanol) 1,4-dioxan solvate, 4C(18)H(16)OSi x C(4)H(8)O(2), (III), the 1,4-dioxan component lies across an inversion centre in space group P-1 and centrosymmetric five-molecule aggregates are linked by paired C[bond]H...pi(arene) interactions to form molecular ladders. PMID- 12094063 TI - A comparative study of intermolecular interactions in the crystal structures of phenyl/phenyl end-capped oligoanilines. AB - Three crystal structures have been analysed from the point of view of intermolecular interactions: N,N'-diphenyl-1,4-benzoquinone diimine, C(18)H(14)N(2), (I), its reduced form N,N'-diphenyl-1,4-phenylenediamine, C(18)H(16)N(2), (II), and N,N'-diphenyl-1,4-phenylenediammonium bis(p toluenesulfonate), C(18)H(18)N(2)(2+) x 2C(7)H(7)O(3)S(-), (III), which contains fully protonated (II) with p-toluenesulfonic acid. The local molecular C(i) symmetry is preserved in all three structures and the packing seems to be dominated by the mutual arrangement of the simple polyaniline oligomers in the different protonation states. In (I), the most significant molecular interactions are stacking forces, forming columns of molecules along [001]. Close packing of the columns results in C-centering of the structure. In (II), only van der Waals interactions can be observed. In the structure of (III), the p-toluenesulfonate ions serve as acceptors in relatively strong N[bond]H...O hydrogen bonds. The N,N'-diphenyl-1,4-phenylenediammonium cation intercalates between two anions related by a centre of symmetry. PMID- 12094065 TI - 4,5-Dihydro-3-methyl-5-(4-methylphenyl)-1H-pyrazole-1-carboxamidinium acetate acetone hemisolvate. AB - The X-ray structure analysis of the unexpected product of the reaction between 4 (4-methylphenyl)but-3-en-2-one and aminoguanidine revealed the title compound, C(12)H(17)N(4)(+) x -C(2)H(3)O(2)(-) x 0.5C(3)H(6)O, consisting of a protonated amidine moiety joined to a substituted pyrazoline ring at the N1 atom. The amidine group is protonated and the positive charge is delocalized over the three C[bond]N bonds in a similar manner to that found in guanidinium salts. The amidinium moiety of the cation is linked to the acetate anions through four N[bond]H...O hydrogen bonds, with N...O distances of 2.749 (4), 2.848 (4), 2.904 (4) and 2.911 (4) A. The pyrazoline ring adopts a flattened envelope conformation and the substituted phenyl ring is oriented perpendicular to the attached heterocycle. The acetone solvate molecule lies across a twofold rotation axis. PMID- 12094064 TI - 16-(4-Cyanobenzylidene)-3 beta-pyrrolidinoandrost-5-en-17 beta-ol monohydrate. AB - In the title compound, C(31)H(40)N(2)O x H(2)O, the outer two six-membered rings are in chair conformations, while the central ring is in an 8 beta,9 alpha-half chair conformation. The five-membered ring adopts a 13 beta-envelope conformation and the cyanobenzylidene moiety has an E configuration with respect to the hydroxyl group at position 17. The steroid nuclei are linked by intermolecular O[bond]H...O and O[bond]H...N hydrogen bonds to form a molecular network. The molecular packing has an interesting feature, with the steroids aligned parallel to the b axis, forming a closed loop through hydrogen bonds linked via water molecules. PMID- 12094066 TI - Ethyl N-(2-benzylamino-6-benzyloxy-5-nitrosopyrimidin-4-yl)glycinate: sheets built from a three-centre N[bond]H...(N,O), and two-centre C[bond]H...O and C[bond]H...pi(arene) hydrogen bonds. AB - In the title compound, C(22)H(23)N(5)O(4), the molecules are linked into chains by a three-centre N[bond]H...(N,O) hydrogen bond, reinforced by a two-centre C[bond]H...O hydrogen bond, and the chains are further linked into sheets by a combination of C[bond]H...O and C[bond]H...pi(arene) hydrogen bonds. PMID- 12094067 TI - N-(2-iodobenzylidene)-3-nitroaniline: sheets formed by a combination of C[bond]H...O hydrogen bonds and two aromatic pi-pi-stacking interactions. AB - Molecules of the title compound, C(13)H(9)IN(2)O(2), are linked into [010] chains by a single C[bond]H...O hydrogen bond and these chains are linked into (100) sheets by two independent aromatic pi-pi-stacking interactions, each involving one of the two substituted arene rings. PMID- 12094068 TI - 3-Amino-1,2,4-triazine. AB - In the crystal structure of 3-amino-1,2,4-triazine, C(3)H(4)N(4), the molecules form hydrogen-bonded chains that are almost parallel to the b axis (3.2 degrees), and which are inclined to the a and c axes by approximately 21 and approximately 69 degrees, respectively. The distortion of the 1,2,4-triazine ring in the crystal is compared with gas-phase ab initio molecular-orbital calculations. PMID- 12094069 TI - 2-(exo-tricyclo[5.2.1.0(2,6)]deca-4,8-dien-3-endo-yl)acetaldehyde 2,4 dinitrophenylhydrazone. AB - The six-membered ring of the norbornene moiety in the title compound, C(18)H(18)N(4)O(4), is in a slightly distorted boat conformation, and the two five-membered rings within it adopt envelope conformations. The structure is stabilized by inter- and intramolecular N[bond]H...O hydrogen bonds. PMID- 12094070 TI - N,N-Diethyl-2,6,6-trimethyl-4-(3-nitrophenyl)-5-oxo-1,4,5,6,7,8 hexahydroquinoline-3-carboxamide. AB - The title compound, C(23)H(29)N(3)O(4), has potential calcium modulatory properties. The conformation of the 1,4-dihydropyridine ring is unusual in that it is planar, instead of the usual shallow boat. The 3-nitrophenyl substituent is in the synperiplanar orientation with respect to the dihydropyridine ring plane. The oxocyclohexene ring has a distorted envelope conformation, with the out-of plane atom being disordered on opposite sides of the ring plane. The molecules are linked into chains by intermolecular hydrogen bonds. PMID- 12094071 TI - The low-temperature phase of di-tert-butylsilanediol. AB - The crystal structure of di-tert-butylsilanediol, C(8)H(20)O(2)Si, has a reversible phase transition at 211 (2) K. The orthorhombic high-temperature structure has space group Ibam, with Z' = 1/2, and shows a disordered hydrogen bonding system. The low-temperature structure, determined at 143 (2) K, has a twinned monoclinic cell, with space group C2/c and Z' = 2, and shows an ordered hydrogen-bonding system. PMID- 12094072 TI - 4-(allylamino)-2-amino-6-benzyloxy-5-nitrosopyrimidine from synchrotron data at 150 K: double chains built from N[bond]H...N, N[bond]H...O, N[bond]H...pi(arene) and aromatic pi-pi-stacking interactions. AB - In the title compound, C(14)H(15)N(5)O(2), the intramolecular dimensions are consistent with a highly polarized electronic structure. The molecules are linked into chains by a combination of N[bond]H...N, N[bond]H...O and N[bond]H...pi(arene) hydrogen bonds, and the chains are linked in pairs by aromatic pi-pi-stacking interactions PMID- 12094073 TI - Molecular characterization of regenerated cardiomyocytes derived from adult mesenchymal stem cells. AB - We recently isolated a cardiomyogenic (CMG) cell line from murine bone marrow stroma, and in this paper characterize regenerated cardiomyocytes derived from adult mesenchymal stem cells at the molecular level. Stromal cells were immortalized, exposed to 5-azacytidine, and repeatedly screened for spontaneously beating cells. CMG cells began to beat spontaneously after 2 weeks, and beat synchronously after 3 weeks. They exhibited sinus-node-like or ventricular-cell like action potentials. Analysis of the isoforms of contractile protein genes, such as of myosin and alpha-actin, indicated that their phenotype was similar to that of fetal ventricular cardiomyocytes. The cells expressed Nkx2.5, GATA4, TEF 1, and MEF2-C mRNA before 5-azacytidine exposure, and MEF2-A and MEF2-D after exposure. CMG cells expressed alpha1A, alpha1B, and alpha1D-adrenergic receptor mRNA prior to differentiation, and beta1, beta2-adrenergic and M1, M2-muscarinic receptors after acquiring the cardiomyocyte phenotype. Phenylephrine induced phosphorylation of ERK1/2, and the phosphorylation was inhibited by prazosin. Isoproterenol increased the cAMP level 38-fold and beating rate, cell motion, %shortening, and contractile velocity by 48%, 38%, 27%, and 51%, respectively, and the increases were blocked by CGP20712A (beta1-selective blocker). Carbachol increased IP3 32-fold, and the increase was inhibited by AFDX116 (M2-selective blocker). These findings demonstrated that the regenerated cardiomyocytes were capable of responding to adrenergic and muscarinic stimulation. This new cell line provides a model for the study of cardiomyocyte transplantation. PMID- 12094074 TI - Radiological protection for diagnostic examination of pregnant women. AB - Application of diagnostic X-ray examination to pregnant women is complicated since risks to both mother and embryo/fetus must be considered. Embryos and fetuses are more sensitive to radiation than adults or children. The threshold doses for fetal death, malformations and mental retardation which are deterministic effects, are reported to be 100-200 mGy or higher. The relative risk for childhood cancer due to radiation at an absorbed dose of 10 mGy during embryonic/fetal development has been estimated at 1.4. However, the absorbed dose of the embryo/fetus during X-ray diagnostic examination in which the X-ray beam does not irradiate the embryo/fetus directly such as maternal skull and chest X ray is extremely low, less than 0.01 mGy. Thus these diagnostic procedures are not a problem from the perspective of radiological protection of the embryo/fetus. However, for pelvic CT scan and barium enema in which the uterus is directly within the X-ray beam, the absorbed doses to the embryo/fetus are about 20-80 mGy and 10-20 mGy, respectively. Therefore, medical staff must pay careful attention to the embryo/fetus in application of these examinations. Pregnant women who were not aware of pregnancy at the time of their diagnostic exposure have great anxiety about radiation from such X-ray examinations. However, fetal doses below 100 mGy should not be considered a reason for terminating a pregnancy. PMID- 12094075 TI - PSK, a biological response modifier, modifies p53 expression, mitosis and apoptosis in X-ray irradiated mouse embryos: possible cellular mechanism of the anti-teratogenic effect. AB - We previously showed that PSK, a biological response modifier, suppressed X-ray irradiation induced ocular anomalies in mouse embryos. In the present study, in mouse embryos irradiated at E7.5, PSK, when administered immediately after irradiation, suppressed mitosis and increased apoptosis as compared with embryos not treated with PSK at 12 hrs after irradiation. In the irradiated embryos, p53, which is normally expressed at a high level in early embryos, increased at 6 hrs and decreased at 12 hrs after irradiation. In the irradiated and PSK-treated embryos, the p53 level did not change at 6 hrs, increased at 12 hrs and decreased at 24 hrs after irradiation. This timing of PSK-induced delayed increase of p53 coincided with that of the PSK-induced decrease in mitosis and increase in apoptosis. These results suggested that PSK modified the p53 level and affected cell proliferation and apoptosis, which might contribute to the suppression of teratogenesis. PMID- 12094076 TI - Chromosome analysis of blastocysts cultured under the diabetic condition. AB - Chromosomes of Slc:ICR mouse blastocysts cultured under the diabetic condition were analyzed to clarify the effect of glucose and ketone body (DL-beta hydroxybutyric acid). In the group exposed to glucose plus ketone body or glucose alone, blastocysts showed higher incidences of chromosome abnormalities, especially numerical abnormalities such as aneuploidy and polyploidy, than in the control group (p < 0.01). The association of nucleolus organizing regions was increased in the blastocysts exposed to glucose plus ketone body, which seems to be related to the increase in numerical abnormalities. Structural abnormalities such as break and fragment were also observed, but there was no significant difference between the diabetic and nondiabetic conditions. These results from chromosome analysis of the cultured blastocysts suggest that the diabetic condition may directly cause chromosome abnormalities in early embryos, especially aneuploidies, and may thus induce duplications or deletions of genes. These chromosomal damages may disrupt the developmental programs for organogenesis and may be involved in diabetes-induced teratogenesis. PMID- 12094077 TI - Hydrocephalus manifestation in the genetic polydactyly/ arhinencephaly mouse (Pdn/Pdn). AB - The genetic polydactyly/arhinencephaly mouse, Pdn/Pdn, exhibits severe polydactyly both in the fore- and hindlimbs, hydrocephalus, and agenesis of the olfactory bulbs, corpus callosum, and anterior commissure. The mechanism of hydrocephalus manifestation in Pdn/Pdn was investigated in the present study. Ink was injected into the left lateral ventricle in the Pdn/Pdn and +/+ newborn mice. After incubation at 32 degrees C for different time intervals, the heads were fixed in Bouin's solution and were subsequently decalcified in 0.5 mol/L of EDTA solution, paraffin sectioned, and stained with hematoxylin and eosin. Ink spread into the 3rd and right lateral ventricles and flowed to the 4th ventricle and Magendie's foramen rapidly in Pdn/Pdn mice. This rapid spread was due to the dilatation of the interventricular foramen and that the lateral ventricle was directly connected with enlarged 3rd ventricle in Pdn/Pdn. In spite of the rapid spread of ink in the cerebrospinal fluid pathway, ink was not observed in the subarachnoid space around the superior sagittal sinus at 3.5 or 10 hours in Pdn/Pdn mice. The superior sagittal sinus was narrower in Pdn/Pdn than in +/+, and the arachnoid villi were not observed in Pdn/Pdn. From these observations, we suggested that absorption of cerebrospinal fluid from the arachnoid villi in the superior sagittal sinus stagnated and that stagnation of the fluid in the ventricles was the cause of hydrocephalus in Pdn/Pdn mice. PMID- 12094078 TI - Postnatal survival of rat offspring prenatally exposed to pure 2,4 dichlorophenoxyacetic acid (2,4-D). AB - In previous report on prenatal developmental effects of pure 2,4 dichlorophenoxyacetic acid (2,4-D) in rat, we found that this chemical was maternally toxic, embryolethal, and that it induced urogenital malformations in the fetuses. In the present report, we investigated the postnatal survival of the offspring prenatally exposed to 2,4-D during organogenesis, to determine the participation of urogenital malformations on postnatal survival. We used doses of 70 mg, 110 mg and 150 mg, which were each found to induce significant urogenital malformations, when administered in different periods of organogenesis: GD 6 to 15, GD 6 to 10, and GD 11 to 15. We found that 2,4-D has a significant influence on progeny viability by increasing the postnatal death. The kidney and urinary tract malformations induced in the fetuses might be the cause of the increased rate of postnatal death. 2,4-D did not impair the postnatal growth of the unaffected offspring. PMID- 12094079 TI - Alteration of androgen receptor immunoexpression by neonatal DES-treatment in mouse epididymis. PMID- 12094080 TI - Asthma research inspires NP practice. PMID- 12094083 TI - Identify bipolar spectrum disorders. AB - Patients with bipolar spectrum disorders commonly present with depressive symptoms to primary care clinicians. This article details bipolar spectrum disorder assessment, treatment, and treatment response. By intervening early in the course of depressive and hypomanic episodes, you can help decrease the morbidity and suffering associated with bipolar spectrum disorders. PMID- 12094084 TI - Update your understanding of influenza. AB - Each year, approximately 20,000 Americans die from influenza infections. Those particularly susceptible include the elderly and patients with chronic medical conditions. Immunoprophylaxis with an inactivated vaccine and chemoprophylaxis with an influenza antiviral drug help minimize influenza incidence. Here, we review the biology behind influenza viruses, clinical manifestations, diagnosis, treatment, and prevention measures, including the Advisory Committee on Immunization Practices' recommendations regarding the influenza vaccine. PMID- 12094085 TI - A look at colorectal cancer screening in women. AB - Aroutine physical examination often includes a digital rectal examination (DRE) and a fecal occult blood test (FOBT). To determine the diagnostic yield of these tests in women younger than age 50, the author performed a literature review. This article reviews the most recent research on the DRE and FOBT, outlines guidelines for colorectal screening, and discusses the efficacy and benefits of these guidelines. PMID- 12094086 TI - Ease symptoms of pulmonary hypertension with bosentan. PMID- 12094087 TI - Are your prescriptions part of a disappearing act? PMID- 12094088 TI - Climate change and infectious disease: stormy weather ahead? PMID- 12094089 TI - After the wall. PMID- 12094090 TI - Genetics and infectious disease: convergence at the prion. PMID- 12094091 TI - On the prevention of preeclampsia: nutritional factors back in the spotlight? PMID- 12094092 TI - Predicting Ross River virus epidemics from regional weather data. AB - BACKGROUND: Diseases caused by arboviruses cause extensive mortality and morbidity throughout the world. Weather directly affects the breeding, abundance, and survival of mosquitoes, the principal vector of many arboviruses. The goal of this study was to test whether climate variables could predict with high levels of accuracy (more than 70%) epidemics of one arbovirus, Ross River virus disease. METHODS: Weather data from two regions in southeastern Australia were matched with Ross River virus disease data for the period 1991 to 1999. Our aim was to develop simple models for the probability of the occurrence of an epidemic in an area in a given year. RESULTS: Two predictable epidemic patterns emerged, after either high summer rainfalls or high winter rainfalls. A prerequisite relating to host-virus dynamics was lower than average spring rainfall in the preepidemic year. The sensitivity of the model was 96% for Region 1 and 73% for Region 2. CONCLUSIONS: Early warning of weather conditions conducive to outbreaks of Ross River virus disease is possible at the regional level with a high degree of accuracy. Our models may have application as a decision tool for health authorities to use in risk-management planning. PMID- 12094093 TI - Improved air quality in reunified Germany and decreases in respiratory symptoms. AB - BACKGROUND: Previous research on air pollution effects has found associations with chronic adverse health effects even at the relatively low levels of ambient particulates currently measured in most urban areas. METHODS: We assessed the impact of declines of total suspended particulates and sulfur dioxide in eastern Germany after reunification on the prevalence of nonallergic respiratory disorders in children. In the 1990s, particle mass (total suspended particulates) and sulfur dioxide declined, whereas number concentrations of nucleation-mode particles (10-30 nm) increased. In three study areas, questionnaires for 7,632 children between 5 and 14 years of age were collected in three phases: 1992-1993, 1995-1996, and 1998-1999. RESULTS: Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for a 50-microg/m3 increment in total suspended particulates were 3.0 (CI = 1.7-5.3) for bronchitis, 2.6 (CI = 1.0-6.6) for sinusitis, and 1.9 (CI = 1.2-3.1) for frequent colds. The effect sizes for a 100-microg/m3 increment in sulfur dioxide were similar. The effect estimates for ambient total suspended particulates and sulfur dioxide were stronger among children not exposed to gas stove emissions, visible molds or dampness, cats, or environmental tobacco smoke. CONCLUSIONS: The decreasing prevalence of nonallergic respiratory symptoms, along with improvements in ambient particle mass and sulfur dioxide (but not in nucleation-mode particles), indicates the reversibility of adverse health effects in children. This adds further evidence of a causal association between combustion-related air pollutants and childhood respiratory symptoms. PMID- 12094094 TI - The incubation period of kuru. AB - BACKGROUND: Kuru is a transmissible spongiform encephalopathy that was identified in Papua New Guinea in the late 1950s. Several thousand cases of the disease occurred during a period of several decades. Epidemiologic investigations implicated ritual endocannibalistic funeral feasts as the likely route through which the infectious agent was spread. METHODS: We estimated the incubation period distribution of kuru using a back-calculation model and explored the relation among sex, age at infection, and incubation period. Key assumptions in the model were that the number of new kuru infections in a year was proportional to the number of kuru cases dying that year, and that the epidemic arose from a single case of sporadic Creutzfeldt-Jakob Disease occurring around 1900. RESULTS: The mean incubation period of kuru was estimated at between 10.3 and 13.2 years. Point estimates of the 90th percentile ranged from 21.1 to 27.0 years. The incubation period in females was estimated to be shorter than that in males. The shortest incubation periods were estimated in adult women, who may have been exposed to the largest doses of infectious material. CONCLUSIONS: Our findings suggest that the relatively young age of cases of variant Creutzfeldt-Jakob disease probably reflects increased levels of exposure in young people, rather than age-dependency in the incubation period. PMID- 12094095 TI - Vitamin C and the risk of preeclampsia--results from dietary questionnaire and plasma assay. AB - BACKGROUND: Oxidative stress plays an important role in the pathophysiology of preeclampsia. METHODS: In a case-control study of 109 women with preeclampsia and 259 controls, maternal dietary and plasma vitamin C in relation to preeclampsia risk were assessed. Dietary intake during the periconceptional period and pregnancy was ascertained using a semiquantitative food frequency questionnaire. Logistic regression procedures were used to derive odds ratios (OR) and 95% confidence intervals (CI). Plasma ascorbic acid was determined using automated enzymatic procedures. RESULTS: After adjusting for maternal age, parity, prepregnancy body mass index, and energy intake, women who consumed <85 mg of vitamin C daily (below the recommended dietary allowance), as compared with others, experienced a doubling in preeclampsia risk (OR = 2.1; 95% CI = 1.1-3.9). The OR for extreme quartiles of plasma ascorbic acid (<42.5 vs > or = 63.3 micromol/liter) was 2.3 (95% CI = 1.1-4.6). Compared with women in the highest quartile, those with plasma ascorbic acid <34.6 micromol/liter (lowest decile) experienced a 3.8-fold increased risk of preeclampsia (95% CI = 1.7-8.8). CONCLUSIONS: Our results, if confirmed, would suggest that current public health efforts to increase intake of fruits and vegetables rich in vitamin C and other antioxidants may reduce the risk of preeclampsia. PMID- 12094096 TI - Perinatal risk factors for infantile autism. AB - BACKGROUND: Etiologic hypotheses in infantile autism suggest a strong genetic component, as well as possible environmental risks linked to early fetal development. We evaluated the association of maternal, pregnancy, delivery, and infant characteristics and risk of infantile autism. METHODS: We conducted a case control study nested within a population-based cohort (all Swedish children born in 1974-1993). We used prospectively recorded data from the Swedish Birth Register, which were individually linked to the Swedish Inpatient Register. Cases were 408 children (321 boys and 87 girls) discharged with a main diagnosis of infantile autism from any hospital in Sweden before 10 years of age in the period 1987-1994, plus 2,040 matched controls. Conditional logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: The risk of autism was associated with daily smoking in early pregnancy (OR = 1.4; CI = 1.1-1.8), maternal birth outside Europe and North America (OR = 3.0; CI = 1.7 5.2), cesarean delivery (OR = 1.6; CI = 1.1-2.3), being small for gestational age (SGA; OR = 2.1; CI = 1.1-3.9), a 5-minute Apgar score below 7 (OR = 3.2, CI = 1.2 8.2), and congenital malformations (OR = 1.8, CI = 1.1-3.1). No association was found between autism and head circumference, maternal diabetes, being a twin, or season of birth. CONCLUSIONS: Our findings suggest that intrauterine and neonatal factors related to deviant intrauterine growth or fetal distress are important in the pathogenesis of autism. PMID- 12094097 TI - Estimating the conditional false-positive rate for semi-latent data. AB - When comparing tests for a disease, it is necessary to know whether individuals are diseased or nondiseased. In practice, the confirmatory (gold standard) procedure is often limited to individuals with positive test results, because the confirmatory procedure is not applied to individuals with negative test results. We present a model for estimating the sensitivity and specificity when two tests are compared and the gold standard classification is unavailable (semi-latent) for those individuals with negative results on both tests. The model does not assume independent error rates, and estimates of specificity conditional on a false-positive result for another test are derived. We use a Bayes approach for estimating the distributions of the performance parameters. PMID- 12094098 TI - Incidence of acute urinary tract infection in young women and use of male condoms with and without nonoxynol-9 spermicides. AB - BACKGROUND: Acute urinary tract infection is one of the most common infections seen in primary care. METHODS: We conducted a nested case-control study among a cohort of 519 women, ages 15-29 years, enrolled in a contraceptive acceptability study to examine whether recent use of male condoms increases urinary tract infection risk. RESULTS: One hundred sixty-five incident urinary tract infections were identified during 12-month follow-up periods in a cohort study that was conducted between 1996 and 1999. After exclusions for urinary tract infection recurrences, pregnancy, antibiotic use, diabetes, diaphragm/cervical cap use, or urinary tract abnormalities, there were 100 cases and 200 controls. Compared with women not using barrier methods (and after adjustment for age, urinary tract infection history, hormonal method use, and frequency of sex) the odds ratio (OR) for any reported use of condoms coated with spermicide (Nonoxynol-9) in the previous 30 days was 2.8 (95% [confidence interval] CI = 1.2-6.5). The OR was 11.5 (95% CI = 2.5-53) for exclusive Nonoxynol-9-coated condom use. The OR for exclusive use of non-Nonoxynol-9-coated condoms was 7.4 (95% CI = 1.6-35). CONCLUSIONS: In this study, use of male condoms was associated with increased urinary tract infection risk; the largest risk was associated with exclusive condom use and use of Nonoxynol-9-coated condoms. PMID- 12094099 TI - Multiple imputation to account for missing data in a survey: estimating the prevalence of osteoporosis. AB - BACKGROUND: Nonresponse bias is a concern in any epidemiologic survey in which a subset of selected individuals declines to participate. METHODS: We reviewed multiple imputation, a widely applicable and easy to implement Bayesian methodology to adjust for nonresponse bias. To illustrate the method, we used data from the Canadian Multicentre Osteoporosis Study, a large cohort study of 9423 randomly selected Canadians, designed in part to estimate the prevalence of osteoporosis. Although subjects were randomly selected, only 42% of individuals who were contacted agreed to participate fully in the study. The study design included a brief questionnaire for those invitees who declined further participation in order to collect information on the major risk factors for osteoporosis. These risk factors (which included age, sex, previous fractures, family history of osteoporosis, and current smoking status) were then used to estimate the missing osteoporosis status for nonparticipants using multiple imputation. Both ignorable and nonignorable imputation models are considered. RESULTS: Our results suggest that selection bias in the study is of concern, but only slightly, in very elderly (age 80+ years), both women and men. CONCLUSIONS: Epidemiologists should consider using multiple imputation more often than is current practice. PMID- 12094100 TI - Lung cancer among rock and slag wool production workers. AB - BACKGROUND: Previous cohort studies have found an elevated risk of lung cancer among rock and slag wool (RSW) production workers. The aim of the present study was to investigate the risk of lung cancer associated with exposure to RSW while controlling for other occupational exposures and tobacco smoking. METHODS: Since 1971, a total of 196 lung cancer cases occurred among men who worked in seven plants in Denmark, Norway, Sweden, and Germany, with start of production between 1937 and 1950. We obtained information on occupational history and extra occupational factors from either the subject or the next of kin for 133 cases and 513 matched controls. We assessed occupational exposure on the basis of interview data combined with information from expert panels set up in each factory. RESULTS: For cumulative exposure to RSW assessed with a 15-year lag, the smoking adjusted odds ratios in the second, third, and fourth quartiles of exposure were 1.3 (95% confidence interval [CI] = 0.7-2.3), 1.0 (CI = 0.5-1.9), and 0.7 (CI = 0.3-1.3). Similar results were obtained when we included only those workers employed for more than 1 year, when we included other indicators of RSW exposure, and after control for co-exposures. CONCLUSIONS: This study provides no evidence of a carcinogenic effect on the lung of rock and slag wool under exposure circumstances in the production industry during the last four to five decades. PMID- 12094101 TI - DDT metabolite and androgens in African-American farmers. AB - BACKGROUND: The ubiquitous dichlorodiphenyltrichloroethane (DDT) metabolite 1,1 dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE) is an androgen receptor antagonist. Data on potential antiandrogenic activity of DDE in humans are limited. METHODS: The relations between concentrations of plasma DDE and several serum androgens (total testosterone, bioavailable testosterone, 5alpha dihydrotestosterone, and free androgen index) were examined in 137 North Carolina black male farmers, using multiple linear regression. RESULTS: Participants ranged in age from 30 to 88 years (mean = 62 years). Most had farmed for about 30 years and 27% reported having used DDT. The median DDE level was 7.7 microg per liter (1213 microg per kg lipid), slightly higher than in other recent studies. Overall, concentrations of DDE and androgens were unrelated. Total testosterone decreased 2% (95% confidence limits [CL] = -9%, 5%) per increase in interquartile distance of lipid-adjusted DDE. The percentage change in other hormones was similarly negligible. However, among those whose DDE level was in the top tenth percentile, compared with all others, total testosterone and free androgen index were lower by 23% (CL= -40%, 1%) and 22% (CL =-41%, 4%) respectively. Plasma androgen levels decreased with age, a relation that has previously been studied only in whites. CONCLUSIONS: Studies of more highly exposed populations may be needed to evaluate effects, if any, of DDE. PMID- 12094102 TI - Estimating attributable fraction in partially ecologic case-control studies. AB - Partially ecologic case-control studies combine group-level exposure data with individual-level data on disease status, group membership, and covariates. If the exposure measure is the exposure prevalence of various groups, the attributable fraction (AF; the estimated proportion of cases that are attributable to exposure) can be estimated by classifying all subjects in groups with exposure prevalence above zero as exposed. Such a threshold AF estimator ([AF]T) is unbiased in confounding-free situations if the threshold is 100% sensitive, but it might be imprecise. We propose an alternative AF estimator, [AF]L, for partially ecologic case-control studies under a linear model for the association between the exposure prevalence and the odds ratio. The proposed estimator can also be applied to situations in which covariate adjustment is necessary. [AF]T and [AF]L are compared with respect to precision and bias. [AF]L is also unbiased when the exposure prevalence is zero in the group(s) assessed as unexposed. Using [AF]L will consistently result in improved precision compared with [AF]T, although the results may not differ substantially. The 95% confidence intervals for both AF estimators show satisfactory coverage in bias-free exposure scenarios. Pronounced negative bias and decreased coverage result for both AF estimators even when small fractions (3-9%) of exposed subjects are included in the group assessed as unexposed. PMID- 12094104 TI - The changing face of epidemiology in the genomics era. PMID- 12094103 TI - Cigarette smoking and the risk of mucinous and nonmucinous epithelial ovarian cancer. AB - BACKGROUND: The association between cigarette smoking and ovarian cancer may vary according to the histologic type of tumor. METHODS: We examined cigarette smoking as a risk factor for both mucinous and nonmucinous tumors in a population-based case-control study comparing 767 incident cases of epithelial ovarian cancer with 1,367 community controls frequency matched to cases by age and race. RESULTS: Smoking was associated with mucinous tumors (odds ratio [OR] = 1.9; 95% confidence interval [CI] = 1.3-2.9) but not with nonmucinous tumors (OR = 1.1; CI = 0.9-1.3). Furthermore, the odds ratios for smokers with mucinous tumors increased with increasing pack-years of smoking (OR = 1.0, 1.9, and 2.7 for <5, 5 24, and > 24+ pack-years, respectively; P for trend = 0.01) CONCLUSIONS: Cigarette smoking appears to be a risk factor for mucinous but not for nonmucinous tumors. PMID- 12094105 TI - Air pollution and asthma among children in Seoul, Korea. AB - BACKGROUND: As information about the health risks associated with air pollution has become available, attention has focused increasingly on susceptible persons such as children and persons with preexisting respiratory diseases, such as asthma. METHODS: We investigated the association between outdoor air pollution and asthma attacks among children under 15 years of age in Seoul, Korea. We estimated the relative risks of hospitalization associated with an interquartile range (IQR) increase in pollutant concentrations and used time series analysis of the counts by means of the generalized additive Poisson model. RESULTS: The estimated relative risk of hospitalization for asthma was 1.07 (95% confidence interval [CI] = 1.04-1.11) for particulate matters less than or equal to 10 microm in aerodynamic diameter (IQR = 40.4 microg/m3); 1.11 (95% CI = 1.06-1.17) for sulfur dioxide (IQR = 4.4 ppb); 1.15 (95% CI = 1.10-1.20) for nitrogen dioxide (IQR = 14.6 ppb); 1.12 (95% CI = 1.07-1.16) for ozone (IQR = 21.7 ppb); and 1.16 (95% CI = 1.10-1.22) for carbon monoxide (IQR = 1.0 ppm). CONCLUSIONS: These findings support the hypothesis that air pollution at levels below the current standards of Korea is harmful to sensitive subjects such as asthmatic children. PMID- 12094106 TI - Maternal fever, multivitamin use, and selected birth defects: evidence of interaction? AB - BACKGROUND: Multivitamin use has been associated with lower risks for some birth defects. We evaluated whether multivitamin use modified birth defect risks associated with febrile illness, a common and possibly teratogenic exposure. METHODS: From the population-based Atlanta Birth Defects Case-Control Study (1968 1980) we selected seven defects (neural tube defects, cleft lip and palate, cardiac outflow tract defects, ventricular septal defects, atrial septal defects, omphalocele, and limb deficiencies) because of their inverse relation with multivitamin supplement use documented in previous analyses. We defined four exposure categories from combinations of multivitamin use (periconceptional use compared with no use) and febrile illness (early pregnancy compared with no illness). The reference category was no multivitamin use and no illness. RESULTS: Febrile illness with no multivitamin use was associated with generally increased risk for the seven defects and the combined group (odds ratio = 2.1, 1.7, 1.5, 1.9, 2.9, 4.4, 3.3, and 2.3, respectively). With multivitamin use, however, the risk estimates associated with febrile illness were generally lower (odds ratio = 0.6, 1.1, 0.0, 1.5, 0.0, 0.8, 0.0, and 0.8, respectively). Some of the associated 95% confidence intervals included one. CONCLUSIONS: The pattern of findings suggests that multivitamin use might decrease the risk associated with febrile illness. PMID- 12094108 TI - Pollution and daily mortality in Amsterdam. PMID- 12094109 TI - No association between major football games and cardiovascular mortality. PMID- 12094110 TI - Sunshine and suicide incidence. PMID- 12094112 TI - Sexual activity and the risk of prostate cancer. PMID- 12094113 TI - General review of maximal aerobic velocity measurement at laboratory. Proposition of a new simplified protocol for maximal aerobic velocity assessment. AB - The maximal aerobic velocity concept developed since eighties is considered as either the minimal velocity which elicits the maximal aerobic consumption or as the "velocity associated to maximal oxygen consumption". Different methods for measuring maximal aerobic velocity on treadmill in laboratory conditions have been elaborated, but all these specific protocols measure V(amax) either during a maximal oxygen consumption test or with an association of such a test. An inaccurate method presents a certain number of problems in the subsequent use of the results, for example in the elaboration of training programs, in the study of repeatability or in the determination of individual limit time. This study analyzes 14 different methods to understand their interests and limits in view to propose a general methodology for measuring V(amax). In brief, the test should be progressive and maximal without any rest period and of 17 to 20 min total duration. It should begin with a five min warm-up at 60-70% of the maximal aerobic power of the subjects. The beginning of the trial should be fixed so that four or five steps have to be run. The duration of the steps should be three min with a 1% slope and an increasing speed of 1.5 km x h(-1) until complete exhaustion. The last steps could be reduced at two min for a 1 km x h(-1) increment. The maximal aerobic velocity is adjusted in relation to duration of the last step. PMID- 12094114 TI - Is velocity-specific strength training important in improving functional performance? AB - A variable considered when designing programs to optimize athletic performance is training velocity. It has been suggested that training at a specific velocity improves strength mainly at that velocity and as velocity deviates from the trained velocity, the less effective training will be. However, the research describing velocity-specific adaptation and the transference of these adaptations to other movement velocities is by no means clear. Compounding the problem in this area is the failure of research to detail the relationship between training velocity and actual movement velocity of a given task or athletic pursuit. In most cases there is a great disparity between training velocity and actual movement velocity. Factors that may better develop and explain velocity-specific adaptation in relation to functional performance are discussed. Developing qualities such as strength, power and rate of force development would appear of greater importance than training at the actual movement velocity of a task. It may be that irrespective of load and limb velocity, the repeated intent to move an isoinertial load as rapidly as possible might be an important stimulus for functional high velocity adaptation. The ability of the nervous system to activate and coordinate agonist, synergist and antagonist activity would seem essential. It was suggested training techniques that simulate the velocity and acceleration profiles associated with the desired functional performance, such as throw or jump training, may optimize functional adaptation. Furthermore combination training that incorporates same session sport specific training with either a heavy load or a mixed training load approach might provide an optimal strategy for promoting intramuscular and intermuscular co-ordination and improving functional performance. PMID- 12094115 TI - Leg strength and stiffness as ability factors in 100 m sprint running. AB - BACKGROUND: The purpose of this study was to determine the importance of leg strength and stiffness relative to i) 100 m sprint performance, ii) mean speed on the three phases of the 100 m race (30-60-100 m) and iii) the speed differences between these phases. METHODS: Nineteen regional to national level male sprinters competed in a 100 m race. Video analysis was used to determine mean velocity parameters. Two subgroups were created since some of the runners decreased their velocity during the third phase (G1), whereas others maintained or accelerated it (G2). Leg strength (concentric half-squats - counter movement jump) and stiffness (hopping) were determined. Simple (r) and multiple regressions (R) were used. RESULTS: The mean performance over 100 m was 11.43 sec (10.72-12.87 sec). The concentric half-squats were related to 100 m (r=0.74, p<0.001) and to the mean speed of each phase (R=0.75, p<0.01). The counter movement jump was related to 100 m (r=0.57, p<0.05) and was the predictor of the first phase (r=0.66, p<0.01). The hopping test was the predictor of the two last phases (R=0.66, p<0.05). Athletes who had the greatest leg stiffness (G1) produced the highest acceleration between the first and the second phases, and presented a deceleration between the second and the third ones. CONCLUSIONS: The concentric half-squats test was the best predictor in the 100 m sprint. Leg stiffness plays a major role in the second phase. PMID- 12094116 TI - Is muscle power related to running speed with changes of direction? AB - BACKGROUND: The purpose of this study was to identify the relationships between leg muscle power and sprinting speed with changes of direction. METHODS: EXPERIMENTAL DESIGN: the study was designed to describe relationships between physical qualities and a component of sports performance. SETTING: testing was conducted in an indoor sports hall and a biomechanics laboratory. PARTICIPANTS: 15 male participants were required to be free of injury and have recent experience competing in sports involving sprints with changes of direction. MEASURES: subjects were timed in 8 m sprints in a straight line and with various changes of direction. They were also tested for bilateral and unilateral leg extensor muscle concentric power output by an isokinetic squat and reactive strength by a drop jump. RESULTS: The correlations between concentric power and straight sprinting speed were non-significant whereas the relationships between reactive strength and straight speed were statistically significant. Correlations between muscle power and speed while changing direction were generally low and non-significant for concentric leg power with some moderate and significant (p<0.05) coefficients found for reactive strength. The participants who turned faster to one side tended to have a reactive strength dominance in the leg responsible for the push-off action. CONCLUSIONS: The relationships between leg muscle power and change-of-direction speed were not consistent. Reactive strength as measured by the drop jump appears to have some importance for lateral change of-direction speed, possibly because of similar push-off actions. It was concluded that reactive strength of the leg extensor muscles has some importance in change-of-direction performance but the other technical and perceptual factors than influence agility performance should also be considered. PMID- 12094117 TI - Effects of lower limbs muscular fatigue on anticipatory postural adjustments during arm motions in humans. AB - BACKGROUND: The purpose of the present experiment was to examine the effects of lower limbs muscular fatigue on the anticipatory postural adjustments during a voluntary arm raising movement. METHODS: Eight adult males (mean age 29+/-6 years) voluntarily participated to the experiment. Subjects' task consisted in performing an unilateral ante-flexion of the right arm over 90 deg, holding a 500 g weight, once having stabilized their upright posture. A similar movement execution was required. Measurements were made in a no-fatigue situation and under a high level of fatigue of lower limbs. Surface EMG of four postural muscles (the ipsilateral and contralateral Gastrocnemius and the ipsilateral and contralateral Semitendinosus) and of one muscle involved in the arm raising (the Anterior Deltoideus of the right arm) was recorded. Body sway was also monitored using a force platform. RESULTS: With fatigue, quantitative EMG analysis showed a decrease of the ipsilateral Semitendinosus activity whereas the temporal EMG analysis showed an increase of its latency with respect to movement onset. CONCLUSIONS: These data suggested a functional adaptation resulting in an invariance of global anticipatory postural adjustments for the two conditions of no-fatigue and fatigue and ensuring an adequate execution of the movement. PMID- 12094118 TI - Influence of changes in nasal ventilation on estimated workload during submaximal field running. AB - BACKGROUND: Breathe Right (BR) external nasal dilator have become increasingly popular over the past few years, however, the physiological basis for using BR remains unclear. We have examined whether alteration in nasal ventilation would influence the metabolic cost of submaximal running. METHODS: Metabolic cost was estimated in 10 male endurance triathletes by measuring heart rate (HR) and exercise perception by measuring the rate of perceived exertion (RPE) during submaximal field running. The protocol consisted of 5 min of running at 80% of their maximal aerobic velocity (MAV(f), previously determined under field conditions) for three randomised experimental conditions, separated by a 10 min rest. The conditions were normal nasal ventilation (N), no nasal ventilation (using a nose clip) (NC) and with a BR. RESULTS: Running with the BR or with NC did not significantly influence HR (N: 173+/-7, BR: 173+/-8, NC: 172+/-7 beat x min(-1); F=0.01, p=0.99) or RPE (N: 12.1+/-1.7, BR: 11.8+/-1.9, NC: 13.2+/-0.8; F=1.88, p=0.18). We conclude that alteration in nasal ventilation produced by using BR or NC do not influence HR or RPE in a group of triathletes running 5 min at 80% of MAV(f). CONCLUSIONS: The present study tended to demonstrate that both nasal ventilation would not influence the total metabolic cost, and that the BR device is not advantageous during high intensity exercise. PMID- 12094119 TI - Jump evaluation of elite volleyball players using two methods: jump power equations and force platform. AB - BACKGROUND: The aim of the present study was to determine the best jump power equation in the evaluation of elite volleyball players using both the force platform and peak power equations. METHODS: Nine elite volleyball players and nine sedentary subjects performed counter-movement jump tests on a force platform. RESULTS: Peak power and height were greater in the volleyball players than in the sedentary subjects, whatever the method used. The results demonstrated that the peak power values obtained on the force platform and those scored from the equations of Lewis, Harman and Sayers et al. were correlated when the whole sample was taken into account. However, a significant equation x level interaction (p<10(-4)) indicated different behaviour as a function of performance level. In sedentary subjects, peak power was significantly underestimated using the Lewis equation (943+/-162 W; p<10(-4)) and did not differ using both the Harman (3004+/-563 W) and Sayers (3400+/-604 W) equations when compared to the peak power noted with the force platform (3372+/-532 W). In contrast, in volleyball players, peak power was underestimated using the three equations (1246+/-78 W, p<10(-4); 4314+/-216 W, p<0.001; 4607+/-251, p<0.005; for the Lewis, Harman and Sayers equations, respectively, versus 5355+/-522 W for the force platform). CONCLUSIONS: The results of the present study demonstrate the difficulty in choosing the most relevant equation in the jump power calculation. PMID- 12094120 TI - Validation of the NFL-225 test for predicting 1-RM bench press performance in college football players. AB - BACKGROUND: The purpose of this study was to evaluate the accuracy of repetitions to-fatigue (RTF) using an absolute load of 102.3 kg (225 lbs) to estimate one repetition maximum (1-RM) bench press performance in college football players using various prediction equations. METHODS: EXPERIMENTAL DESIGN: a prospective study on the association between muscular endurance and muscular strength. PARTICIPANTS: 260 players from NCAA Division IA (n=43), IAA (n=63), II (n=129), and red-shirts (n=25) were evaluated at the conclusion of a minimum of eight weeks of heavy-resistance training during the off-season. MEASURES: all subjects performed a 1-RM bench press and RTF using an absolute load of 102.3 kg. RESULTS: The Mayhew et al. NFL-225 equation nonsignificantly overestimated 1-RM from RTF by 0.5 kg, while the Chapman et al. NFL-225 equation significantly underpredicted by 3.2 kg, although both equations were comparable in the number of players predicted within +/-4.5 kg of actual 1-RM (52% vs 51%, respectively). Only two of nine RTF equations currently in use produced predicted 1-RM values that were not significantly different from actual 1-RM performance. CONCLUSIONS: Specific NFL 225 equations are more accurate in estimating 1-RM bench press from absolute muscle endurance in college football players than previous published RTF equations. The accuracy of prediction decreases at higher repetitions. PMID- 12094121 TI - Physiological factors in middleweight boxing performance. AB - BACKGROUND: We examined, within the middleweight class, the relationship between ranking in boxing competition performance and some physiological factors. METHODS: Eight elite Italian amateur boxers (first series of AIBA ranking) were assessed in 2 testing sessions, a week apart. In the first testing session all subjects underwent anthropometric measurements from which body fat percentage, upper arm and forearm muscle cross-sectional areas were estimated. In the second testing session all subjects performed grip strength measures and a maximal treadmill test to assess oxygen consumption (VO2), blood lactate and heart rate at maximal effort, at individual anaerobic threshold, and at individual ventilatory threshold. The athletes were ranked following the criteria of world amateur AIBA ranking. In this ranking the first ranked boxer had the highest score gained participating in international tournaments. RESULTS: A Spearman rho correlation analysis revealed that the VO2 at individual anaerobic threshold (46.0+/-4.2 ml x kg(-1) x min(-1), r=0.91) and the hand-grip strength (58.2+/-6.9 kg, r=0.87) were highly related (p<0.01) to boxing competition ranking. VO(2max) (57.5+/-4.7 ml x kg(-1) x min(-1), r=0.81) and wrist girth (17.6+/-0.6 cm, r=0.78) were moderately (p<0.05) related. CONCLUSIONS: These data suggest that there are two basic factors related to boxing performance: physical fitness as indicated by individual anaerobic threshold and maximal oxygen consumption, and upper-body muscular strength as indicated by hand-grip strength. PMID- 12094122 TI - Physiological characteristics of Singapore national water polo team players. AB - BACKGROUND: The purpose of this study is to describe the physiological characteristics of members of the Singapore national water polo team. METHODS: EXPERIMENTAL DESIGN: a descriptive study on the physiological characteristics of members of a winning water polo team. SETTING: the study was conducted at the Sports Medicine and Research Centre. PARTICIPANTS: 13 male players (22.5+/-7.2 yrs; 178.5+/-3.9 cm and 71.0+/-8.4 kg). MEASURES: players completed body fat measurements, a submaximal cycling test to determine aerobic fitness and all-out maximal efforts (of arm-cranking and cycling) for 10 and 30 sec duration to determine anaerobic peak power, alactic and lactic acid work capacity for the upper and lower body. RESULTS: The physical measurements were compared with data of national players of various countries. The exercise test results were compared with available published data. CONCLUSIONS: Compared with elite water polo players from other countries, the Singapore water polo players are smaller in stature with a lower body mass. Relative to the limited published data, they are shown to possess a high level of aerobic fitness and anaerobic power and capacity for short-term work. PMID- 12094123 TI - Pre-exercise oral creatine ingestion does not improve prolonged intermittent sprint exercise in humans. AB - BACKGROUND: This investigation determined whether pre-exercise oral Cr ingestion could enhance prolonged intermittent sprint exercise performance. METHODS: EXPERIMENTAL DESIGN: a randomised, double-blind crossover design was employed. SETTING: testing was performed at the Western Australian Institute of Sport and participants were monitored and treated by both scientific and medical personnel. PARTICIPANTS: eight active, but not well-trained males with a background in multiple-sprint based sports acted as subjects for this investigation. INTERVENTIONS: subjects ingested either 15 g Cr.H2O or placebo 120 min and 60 min prior to the start of an 80-min maximal sprint cycling task (10 sets of multiple 6-sec sprints with varying active recoveries). Subjects were retested 14 days later, being required to ingest the alternate supplement and repeat the exercise test. MEASURES: performance variables (work done and peak power) were obtained throughout the exercise challenge. Muscle biopsies (vastus lateralis) were raised to a peak of 2348+/-223 micromol x l(-1) prior to the commencement of exercise after Cr ingestion. There were no significant changes in any cycling performance parameters following Cr ingestion, although blood La- was significantly lower (p<0.05) than placebo at all time points during were taken preexercise as well as immediately and 3 min post-exercise in order to determine concentrations of ATP, PCr, Cr, La- and glycogen. Venous blood was drawn prior to and on four occasions during the exercise test, and analysed for Cr, NH3+, La- and pH. RESULTS: Serum Cr concentrations exercise, and plasma NH3+ accumulation was also significantly reduced (p<0.05) in the Cr condition, but only in the second half of the 80-min exercise test. Muscle ATP and TCr levels as well as postexercise PCr replenishment were unaffected following Cr administration. CONCLUSIONS: The data suggest that although the pre-exercise ingestion of a large Cr dose was shown to have some impact on blood borne metabolites, it does not improve maximal prolonged intermittent sprint exercise performance, possibly due to an insufficient time allowed for uptake of serum Cr by skeletal muscle to occur. Therefore, this form of loading does not provide an alternative method of Cr supplementation to the traditional five-day supplementation regimes established by previous research. PMID- 12094124 TI - Effects of creatine supplementation during recovery from rapid body mass reduction on metabolism and muscle performance capacity in well-trained wrestlers. AB - BACKGROUND: The aim of the study was to test the hypothesis that creatine supplementation with concomitant carbohydrate ingestion during recovery period after rapid body mass reduction accelerates the restoration of body mass and physical performance in well-trained wrestlers. METHODS: A double-blind, placebo controlled cross-over study was conducted on five young healthy male wrestlers, who reduced their body mass by 4.5-5.3% in two series of investigations separated by one month. During 17 hrs recovery period they consumed controlled diet supplemented in random order with glucose (GL trial) or with glucose plus creatine (GL+CR trial). The capacity of the subjects to perform submaximal and maximal (W(max)) intensity work was measured using 5 min intermittent intensity test exercise at the Cybex II device before (Test 1) and after body mass loss (Test 2), also after the recovery (Test 3) on both trials. RESULTS: There was no effect of treatment on the extent of body mass regain during 17 hrs recovery. A significant increase (19.2%) in W(max) from Test 2 to Test 3 was observed in GL+CR trial whereas no change was evident with GL treatment. A strong correlation (r=0.92) was established between the whole body creatine retention and the extent of change in W(max) from Test 2 to Test 3. CONCLUSIONS: The results suggest that creatine supplementation with concomitant glucose ingestion during 17 hrs recovery from rapid body mass loss does not accelerate the restoration of body mass but still stimulates the regain of physical performance in maximal intensity efforts in well-trained wrestlers. PMID- 12094125 TI - Effects of high-calorie supplements on body composition and muscular strength following resistance training. AB - BACKGROUND: Seventy-three healthy, male subjects randomly divided into 3 groups participated in a study to determine the effects of 2 high-calorie nutritional supplements on body composition, body segment circumferences, and muscular strength following a resistance-training (RT) program. METHODS: In addition to their normal diets group 1 (CHO/PRO; n=26) consumed a 8.4 Mj x day(-1) (2010 kcal) high calorie, high protein supplement containing 356 g carbohydrate and 106 g protein. Group 2 (CHO; n=25) consumed a carbohydrate supplement that was isocaloric with CHO/PRO. Group 3 (CTRL; n=22) received no supplement and served as a control. All subjects were placed on a 4-day x week(-1) RT program for 8 weeks. RESULTS: Dietary analysis revealed no significant differences in total energy consumption or nutrients at any time in the non-supplemented diets of the 3 groups. Significant (p= or <0.05) increases in body mass (BM) and fat-free mass (FFM) were observed in CHO/PRO and CHO compared to CTRL. Mean (+/- SD) increases in BM were 3.1+/-3.1 kg and 3.1+/-2.2 kg, respectively. Fat-free mass significantly (p= or <0.05) increased 2.9+/-3.4 kg in CHO/PRO and 3.4+/-2.5 kg in CHO. Muscular strength, as measured by a one-repetition maximum in the bench press, leg press, and lat-pull down increased significantly (p= or <0.05) in all groups. No significant differences in strength measures were observed among groups following training. CONCLUSIONS: Results indicate that high-calorie supplements are effective in increasing BM and FFM when combined with RT. However, once individual protein requirements are met, energy content of the diet has the largest effect on body composition. PMID- 12094126 TI - Video analysis of blows to the head and face at the 1999 World Taekwondo Championships. AB - BACKGROUND: Limited research has been done on head blows that may result in mild traumatic brain injury in Taekwondo. The purpose of this study was to investigate the fighting conditions under which blows to the head commonly take place, with a view to determining the typical conditions under which injury may occur. METHODS: EXPERIMENTAL DESIGN: videotape analysis (retrospective). SETTING: the semi-final and final matches (a total of 48 matches) at the 14th World Taekwondo Championships in 1999. PARTICIPANTS: 64 athletes (32 females and 32 males) who won elimination-round matches (out of 563 competitors), aged 15 to 38 years. MEASURES: frequency, mechanism of head blows, characteristics of situations leading up to and following head blows, frequency of multiple impacts. RESULTS: A total of 35 incidents of head blow occurred (365 blows per 1,000 athlete exposures). All of these head blows were associated with a direct head or face contact and frequently involved: a closed sparring stance, shorter athletes, axe or roundhouse type kicks, attacker's offensive kick, and head-blow-receiver's offensive action with absence of a blocking skill. CONCLUSIONS: To prevent possible brain injury resulting from direct head blows, updated safety education, a complete understanding of concussion for athletes, coaches, and referees, and a rule change in competition Taekwondo are recommended. PMID- 12094127 TI - Combating drug use in competitive sports. An analysis from the athletes' perspective. AB - BACKGROUND: Doping has developed into a widespread problem in competitive and high-performance sports due to increasing professionalism in, and commercialization of sports. In contrast, governments and sports organizations have limited financial resources to support all competitive sports. Therefore, further improvement of anti-doping measures can only be achieved through the inclusion and active participation of the athletes themselves. METHODS: In this study, 101 German athletes who are subject to national and international anti doping tests were asked if doping in sports should be combatted, and which anti doping measures appeared effective from an athlete's perspective. RESULTS: Ninety eight point zero two per cent of those questioned felt that measures should be taken against doping in sports. Improved methods of detection and more information on the health risks were favored, as opposed to more severe punishments. In addition, more than two thirds of the athletes supported the introduction of an anti-doping law. The desire for more frequent drug testing was also expressed, despite the distinct invasion of the athletes' privacy. CONCLUSIONS: An anti-doping law, as requested by the athletes, should include measures for educating the public about the health risks involved with doping. In addition, such a law would also make it possible to develop suitable methods of detection. PMID- 12094128 TI - Application of the Transtheoretical Model to physically active adults. AB - BACKGROUND: A physically activity lifestyle has established health benefits, but interventions to increase exercise adherence have had mixed success. Examining physically active individuals could provide insight into strategies that are effective for exercise maintenance. The purpose of this study was to compare active adults based on exercise stage classification [action (ACT) and maintenance (MT)] according to the Transtheoretical Model (TTM). Physiological and psychosocial differences between stages were hypothesized to be consistent with theory and previous research. METHODS: A cross-sectional design was used to examine components of the TTM, exercise behavior, resting cardiovascular variables, and VO(peak). Data from 57 physically active college students (age= 21.2 +/- 3.7 yrs, 67% female, 71.9% Caucasian) were included in the analysis. TTM variables and self-report of behavior were measured with established questionnaires. VO(peak) was determined from expired gases during a maximal exercise test. RESULTS: Volume of weekly exercise and number of months consistently active were significantly greater for MT (n=35) than for ACT (n=22). When controlling for gender effects, VO(peak) and systolic blood pressure were also greater in MT than ACT. MT also scored higher than ACT on decisional balance (pros-cons of exercise) and 4 processes of change. CONCLUSIONS: Membership in MT and ACT stages was corroborated by aerobic capacity. Predicted differences in TTM components were only partially supported, but behavioral strategies were used more by participants who were consistently active longer. Physically active individuals should be studied over time to determine if targeting behavioral processes of change will enhance long-term exercise adherence. PMID- 12094129 TI - Exercise under hot conditions: a major threat to the immune response? AB - It seems likely that the disturbances of immune response induced by prolonged competitive exercise are exacerbated if athletes also face the stress of hot environmental conditions. We have investigated this question by manipulating the exercise-induced increases of body temperature in a climatic chamber and by submersion of exercisers in a large water-bath. Hot conditions increase the stress of a given bout of exercise, as assessed by personal perceptions, objective (heart rate variability) measures of autonomic nerve balance, and the secretion of "stress" hormones, with a parallel increase in effects upon critical lymphocyte subsets. Changes in the immune response show substantial correlations with plasma concentrations not only of epinephrine (which modulates the adhesiveness of peripherally sequestered lymphocytes), but also with norepinephrine. The latter hormone may mobilize leukocytes from the spleen and lymph glands, or it may act by increasing cardiac output and thus intravascular shear forces. Given the cumulative impact of various environmental stressors upon the immune system, every effort should be made to minimize the athlete's exposure to stresses other than the exercise to be performed. In some circumstances, the use of medications to reduce the overall stress response may also be warranted. PMID- 12094130 TI - TT virus. A review of the literature. AB - In 1997, a new DNA virus was cloned by a Japanese team and designated TT virus (TTV). This virus seemed to be associated with non-A, non-G post-transfusion hepatitis. It was isolated by polymerase chain reaction (PCR) and was presumed to be human Circoviridae. The virus is heterogenous; 16 different genotypes are currently registered, and it can be classified as a "swarm" of at least 5 different viruses. Depending on the PCR technique used, the prevalence of infection ranges from 1.9 to 36% among blood donors, from 11.5 to 71% in hemodialysis patients, from 47 to 82% among patients with non-A, non-B or non-C fulminant hepatic failure, and the most elevated percentage is found in hemophiliacs. Epidemiological studies have established that the routes of TTV infection might be parenteral, oral-fecal, and possibly salivary. Mother-to infant transmission is controversial. TTV may play a role in the pathogenesis of non-A, non-B or non-C fulminant hepatic failure. Patients co-infected with hepatitis C virus (HCV) and TTV have a significantly higher histological grade score than patients with isolated HCV infection. Treatment with interferon seems to decrease TT viremia, according to results obtained outside the context of clinical trials. TTV seems to be a light pathogenic virus. Its widespread presence in the blood of infected subjects contrasts with the apparent absence of pathological symptoms. PCR standardization is needed to clearly establish its real prevalence worldwide. PMID- 12094131 TI - Gram-positive bacterial resistance. A challenge for the next millennium. AB - Penicillin G was first used in 1941. Since then, the trend in bacterial infections has changed. New antibiotics have been developed and bacterial resistance has spread as a consequence. The spread of Gram positive resistant bacteria is related to an inappropriate use of antibiotics. Antibacterial agents are abused or overused in various fields: medicine itself, veterinary science and zootechnics. Now, at the beginning of the third millennium we have been forced to limit our therapeutic options in order to combat these insidious enemies. Selective antibiotic pressure on the microbial population, notably on enterococci and staphylococci, made these two pathogens recalcitrant to traditional chemotherapy. It is a matter of concern that today, vancomycin-resistant Enterococcus spp. (VRE) and vancomycin-intermediate and resistant Staphylococcus aureus (VISA and VRSA) are now being observed worldwide among emerging pathogens. Most pharmaceutical companies are today developing antimicrobial drugs that are active against Gram-positive bacteria. Quinupristin/dalfopristin and linezolid are the most promising drugs and are available only for serious infections; future agents being developed for multi-resistant Gram-positive infections include daptomycin and the glycyclines, although these are still in the development phase. Nevertheless, our group has had the opportunity to treat some serious infections with these drugs and the good results achieved are reported in this review. PMID- 12094132 TI - Clinical role of positron emission tomography (PET) in endocrine tumours. AB - Positron emission tomography (PET) is considered one of the most effective nuclear medicine imaging modality in oncology. In many types of malignant tumours PET has proven to be high sensitive in detecting both primary neoplasm and metastatic disease both before therapy for staging purposes and after surgery, radiotherapy and chemotherapy during follow-up. In particular, PET showed high accuracy in differentiating post-treatment scar from viable tumoural tissue and high sensitivity in visualising very small metastatic foci as those in normal sized (less than 1 cm in diameter) lymph nodes. A large variety of positron emitter tracers have been produced for PET imaging: the most widely used tracer in clinical oncology is 18F-fluoro-2-deoxy-D-glucose (18F-FDG). Deoxy-glucose (DG) is a glucose analog that enters the cell using specific transmenbrane carrier proteins (especially GLUT-1). Once within the cytoplasm, DG is phosphorylated to FG-6-phosphate but does not appear to be further metabolised. In most malignant tumours there is an increase of the glycolityc metabolism which accounts for an increase of the FDG uptake. However, the metabolic cellular activity can be only slightly increased or even normal in well differentiated and slow growing tumours as some endocrine neoplasms. In the present review, the clinical usefulness and limits of PET imaging are discussed in the management of patients with endocrine tumors. PMID- 12094133 TI - The role of hematopoietic stem cells in the treatment of ovarian cancer. AB - In recent years hematopoietic stem cells (HSC) have been the object of new research efforts and scientific advances. Therapeutic strategies have been set up using HSC for the treatment of solid tumors such as ovarian cancer. In this context different approaches have been proposed and clinically investigated. The "autologous" approach refers to the use of HSC as hematologic support to high dose chemotherapy regimens, and to the use of HSC as an abundant source of dendritic cells for cancer vaccination protocols. Our institution has developed a long-term experience in high-dose chemotherapy with autologous HSC transplantation as first-line treatment of advanced ovarian cancer, and in the use of cytokines both for the HSC collection and for the post-transplantation hematopoietic recovery. Moreover, the "allogeneic" approach with HSC consists of the allogeneic transplantation with both myeloablative/standard or nonmyeloablative/reduced conditioning regimens, which has been proposed as a new adoptive immunotherapeutic treatment for different nonhematologic malignancies. Perspectives in the use of HSC in oncology comprise the possibility of an HSC ex vivo expansion, the use of umbilical cord blood HSC, and the development of future HSC-based gene-therapy programs. PMID- 12094134 TI - Treatment-related death from chemotherapy and thoracic radiotherapy for advanced cancer. AB - Pneumonia and sepsis during neutropenia are common complications from chemotherapy and they are sometimes lethal. Radiation pneumonitis is also a common toxicity after thoracic radiotherapy, and severe pneumonitis causes hypoxia and death. However, the incidence and risk factors of treatment-related death from the treatment of advanced cancer using chemotherapy and/or thoracic radiotherapy are not well understood. Between July 1992 and December 1997, 1799 patients were diagnosed as having lung cancer and 784 of 1799 patients received chemotherapy in the National Cancer Center Hospital East. Of 784 patients, 18 (2.3%) died from toxicity of the initial chemotherapy. In the Japan Clinical Oncology Group (JCOG) trials for lung cancer, 29 of 1176 patients (2.5%) were reported to have died from toxicity of the treatments. Several papers have reported the risk factors of early death after chemotherapy or treatment-related death caused by chemotherapy in patients with cancer. Only poor performance status was a vigorous risk factor. Reported mortality rates of radiation pneumonitis range from 0 to 9.9% from 7 reports and 29 of 1244 patients (2.3%) died of pneumonitis after thoracic radiotherapy or chemoradiotherapy. Our previous report suggested that pulmonary fibrosis identified on plain chest X-ray film is a very strong risk factor of treatment-related death from radiation pneumonitis. At least 1-2% mortality should be expected for chemotherapy and thoracic radiotherapy. And in patients with poor PS, the expected mortality rate from chemotherapy is increased. PMID- 12094135 TI - Prostatic intraepithelial neoplasia and prostate cancer. AB - Prostatic intraepithelial neoplasia (PIN) is composed of dysplastic cells with a luminal cell phenotype, expressing the androgen receptor as well as prostate specific antigen. PIN is characterized by progressive abnormalities of phenotype which are intermediate between normal prostatic epithelium (NP) and cancer, indicating impairment of cell differentiation and regulatory control with advancing stages of carcinogenesis. High-grade PIN is considered the most likely precursor of prostatic carcinoma (PCa), according to virtually all available evidence. Androgen deprivation decreases the prevalence and extent of PIN and the degree of capillary vascularization (e.g., angiogenesis) in the surrounding stroma via the suppression of vascular endothelial growth factor (VEGF) production. It is likely that PCa might also arise from precursor lesions other than high-grade PIN (low-grade PIN, atypical adenomatous hyperplasia, malignancy associated foci, and atrophy). PMID- 12094136 TI - New antiepileptic drugs in childhood. AB - In recent years, many new antiepileptic drugs have been available for the treatment of epilepsy in pediatric age. In this review, we describe these new antiepileptic drugs (vigabatrin, lamotrigine, felbamate, gabapentin, tiagabine, topiramate) which can be used together with "old" antiepileptic drugs (e.g. phenobarbital, valproate, etc.). We report their mechanisms of action, effectiveness, dosage and side effects; moreover, some practical advice for their use in pediatric patients is given. PMID- 12094137 TI - Surgery for carcinoma of the gallbladder. Our experience. AB - BACKGROUND: Carcinoma of the gallbladder is a gastrointestinal malignancy with a very poor prognosis. The 5-year survival rate amounts to less than 5% in most series. In this study we reviewed the results of surgical treatment for gallbladder carcinoma with special reference to extended radical procedures. METHODS: Between 1995 and 2000 we enrolled 36 patients (17 males and 19 females), 24 of whom were treated with simple cholecystectomy and 12 with radical resection (partial hepatectomy, regional lymphadenectomy, and common bile duct resection). The tumours were classified by stage using the criteria of the American Joint Committee on Cancer (AJCC). Stages, operative procedures, results of pathologic examinations and the outcome of the resected cases were reviewed. RESULTS: There were 2 postoperative deaths (0.55%). The mean follow-up period was 19.1 months (range 1-60). For stage I and II disease extended cholecystectomy had a better result than simple cholecystectomy: the 5-year survival rates were 38.4 versus 19%, respectively. For the patients with advanced stage III or IV gallbladder carcinoma, a significant advantage of survival resulted in case of liver resection as compared to surgical treatment without liver resection: the 5-year survival rates were 20 and 0%, respectively. CONCLUSIONS: The survival of stage I II patients was good. For the patients in higher stages the prognosis was significantly worse. In these cases more aggressive surgery may be needed. PMID- 12094138 TI - The uncertain primary diagnosis of inflammatory bowel diseases. AB - BACKGROUND: The purpose of this study is to re-evaluate our series of patients affected by a colonic non-neoplastic disease, in order to measure the percentage in whom we were unable to make a correct diagnosis after the first clinical and histological approach and to single-out the reasons for our inability to reach the correct diagnosis. METHODS: During the period 1985-1999 we observed 1228 patients affected by chronic inflammatory colonic diseases. RESULTS: In 859 patients (69.9%) an ulcerative colitis was diagnosed for the first time, and 248 patients (20.1%) were affected by Crohn's colitis. One hundred and twenty-one patients (9.8%) were defined as being affected by an undetermined colitis. Forty three patients of these had a definite diagnosis, afterwards: 27 patients were affected by ulcerative colitis and 16 by Crohn's colitis. Differential diagnosis between inflammatory large bowel diseases (ILBD) and other forms of colitis was set out as follows: 62 cases out of 1228 were consequent on a bacterial infection or parasitosis; in 28 patients a colitis pseudomembranosus was diagnosed. Eighteen cases of ischemic colitis are reported and 14 patients were affected by NSAID-related colitis. In another 6 patients we diagnosed a postradiation colitis. In 22 cases mimicking a Crohn's colitis we ascertained 9 patients affected by intestinal lymphoma, 11 mycobacterium tuberculosis related intestinal infections and 2 cytomegalovirus related colitis. CONCLUSIONS: Despite progress in scientific acquisitions and in diagnostic methods, correct initial diagnosis of ILBD is still difficult, even though it will be defined with time. PMID- 12094139 TI - The "submerged" connective tissue diseases. AB - BACKGROUND: Many patients present with laboratory abnormalities suggestive of connective tissue diseases (CTD), but with a few clinical symptoms, therefore not fulfilling the criteria for a specific diagnosis. This fact led us to suppose the existence of a changeable long "preclinical" phase in CTD development. In the past, we have already dealt with this subject, but the existence of a few reports underlining this concept led us to describe some "paradigmatic" clinical cases, and we suggest the definition of "submerged connective tissue diseases (Sub CTD)". METHODS: During the last 5 years, we observed 25 patients (23 females and 2 males), with a mean age of 32 years, who were enrolled in a prospective study, with a mean follow-up of 23 months. All the patients were ANA-positive and went to our observation presenting only 1 clinical symptom and other laboratory abnormalities suggestive of CTD. RESULTS: The following development of the clinical picture allowed us to make a specific CTD diagnosis in 22 cases. The follow-up study shows the correlation between the starting symptoms, the serological markers (detected at the first examination) and the definitive diagnosis. CONCLUSIONS: From this point of view, we emphasize the importance of the detection of very sensitive and specific serological markers, which are useful for an early diagnosis, particularly in this "submerged" phase of the disease. PMID- 12094140 TI - Phosphate measurements during hypokinesia and phosphate supplements in disclosing phosphate changes in hypokinetic subjects. AB - BACKGROUND: Hypokinesia (diminished movement) induces significant phosphate (P) change, however, little is known about P retention and P depletion during hypokinesia (HK). Measuring P retention and P balance during HK and P supplementation, the objective of this work was to disclose whether HK could contribute to the decreased P retention and consequently to P depletion in normal subjects. METHODS: Studies were done during 30 days pre-HK period and 364 days HK period. Forty normal male individuals aged, 25.3+/-6.4 years were chosen as subjects. They were divided equally into 4 groups: unsupplemented active control subjects (UACS), unsupplemented hypokinetic subjects (UHKS), supplemented active control subjects (SACS) and supplemented hypokinetic subjects (SHKS). Hypokinetic subjects were limited to an average walking distance of 0.5 km day-l, while active control subjects were kept on an average running distance of 9.6 km day-l. Both, SHKS and SACS received daily 14 mmol dicalcium phosphate per kg body weight. RESULTS: Negative P balance, fecal P, urinary calcium (Ca) and P excretion, serum P and total (Cat) level increased significantly (p<0.05) while P retention, serum intact parathyroid hormone (iPTH), 1,25 dihydroxyvitamin D (1,25 (OH)2 D3) and thyrocalcitonin (TC) decreased significantly (p<0.05) in SHKS and UHKS compared with their pre-HK values and their respective active controls (SACS and UACS). However, negative P balance, P retention incapacity, serum, fecal and urinary P level increased significantly (p<0.05) more in SHKS compared with UHKS. Fecal P loss, urinary P and Ca loss, serum P and Cat level, iPTH, TC and 1,25 (OH)2 D3 level, P retention and P balance change insignificantly (p>0.05) in SACS and UACS compared with their pre-HK level. CONCLUSIONS: It was concluded that a significant P excretion in urine and feces in spite of negative P balance and P supplementation may demonstrate reduced P retention, while a significant increase of negative P balance may demonstrate P depletion. Clearly, P intake, regardless of its low or higher dose, was significantly wasted during HK probably due to the decreased ability of the body to retain P. PMID- 12094141 TI - Seroprevalence study of tick borne encephalitis in Turin province. AB - BACKGROUND: Tick borne encephalitis (TBE) virus is diffused in some European countries and it is transmitted by tick bites. In Italy Isodex ricinus represents the main vector of the infection, that rarely produces the neurologic manifestations, characterising the secondary phase of the same. METHODS: In Italy TBE has been little studied and this only in the Middle and Northern regions of the country. Seroepidemiological researches were done prevalently on subjects at high risk of tick bite, such as hunters or forest guards and especially in Trentino and Tuscany. No precise information about TBE virus diffusion was disposable in the Piedmont region and particularly in the Susa valley where, before our investigation failed the data about it. RESULTS: We found that usual hunters and wild boar breeders seem to be particularly exposed to the risk of TBE virus infection, but none neurologic involvement was detected in the anamnesis of the significantly seropositive subjects and also of the borderline ones, that we have studied, despite the limited number of these subjects. CONCLUSIONS: Nevertheless we hope for a following extension of our case report, also in consideration that rare cases of encephalitis of unknown etiology, are signalled in Piedmont. PMID- 12094142 TI - Reliability of bronchoalveolar lavage in the routine clinical assessment of patients with sarcoidosis. A retrospective analysis. AB - BACKGROUND: The aim of the study was to evaluate the reliability of bronchoalveolar lavage in the routine clinical assessment of patients with sarcoidosis. METHODS: STUDY DESIGN: retrospective analysis. SETTING: 2 Divisions of Respiratory Medicine, both in the context of a public general hospital (Ospedale Morelli, Sondalo, Italy and Ospedale Civile, Piacenza, Italy). PATIENTS: 89 patients with definitely diagnosed diffuse infiltrative lung disease, who underwent bronchoalveolar lavage as part of their initial diagnostic work-up in the period 1991-2001. Fifty-one of them had biopsy-proven sarcoidosis. Diagnostic procedures were: transbronchial lung biopsy (n=26), bronchial biopsy (n=11), skin biopsy (n=6), surgical lung biopsy (n=4), superficial lymph node biopsy (n=3), mediastinal lymph node biopsy (n=1). In 38 patients various interstitial lung diseases were diagnosed, according to established criteria. MEASURES: we compared the percentage of BAL lymphocytes and CD4/CD8 ratio in the 2 groups. Sensitivity, specificity, positive and negative predictive value of BAL parameters were calculated with a cut-off value of 28% for BAL lymphocytes and 3.5 for CD4/CD8 ratio. RESULTS: The percentage of lymphocytes was similar in the 2 groups (p=0.19); CD4/CD8 ratio was 5.7+/-4.9 in sarcoidosis and 2.0+/-3.3 in the other patients (p=0.0001). BAL CD4/CD8 ratio in sarcoidosis had sensitivity 58%, specificity 86%, positive predictive value 85%, negative predictive value 61%. CONCLUSIONS: BAL has a low sensitivity in patients with sarcoidosis; CD4/CD8 ratio >or=3.5 strongly suggests sarcoidosis but is not specific enough if considered alone. PMID- 12094143 TI - Vitamin E and carotene status in pre-eclamptic pregnant women from India. AB - BACKGROUND: The imbalance between oxidative stress and the protective antioxidant system of the body enhanced the free radical mediated membrane lipid peroxidation and possibly the vascular endothelial damage due to peroxidation plays a major role in the aetiology of pre-eclampsia. With present day awareness on micronutrient antioxidants, we did investigate vitamin E and carotene status in Indian pre-eclamptic pregnant and full term normotensive pregnant women. Fresh vegetables and oils are considered to be good sources of vitamin E and carotene. The subjects were used to have good intake of fresh vegetable and oil as per Indian standard prescribed by Indian council of Medical research (ICMR) for this sub-continent. METHODS: The blood samples were processed for RBC vitamin E, serum carotene and serum cholesterol analysis. Routine laboratory tests like hemogram, serum urea, urate, malonyldialdehyde, urine sugar and albumin were performed. RESULTS: All pregnant subjects, both cases and control were maternal and gestational age matched. Routine check up showed no significant differences in means of white blood cell count, Hb/hematocrit and platelets. Serum urate and malonyldialdehyde were significantly raised in pre-eclamptic cases. The severely affected pre-eclamptic cases (diastolic BP >100 mmHg with proteinuria 2+ and more) showed markedly low levels of vitamin E and carotene whereas their levels were comparable between mild cases (diastolic BP <100 mmHg with+/-trace albuminuria) and normotensive pregnant control. CONCLUSIONS: The study concluded that the levels of vitamin E and carotene were markedly lowered in severe pre eclamptic pregnant women from India. The mild pre-eclamptic cases did not show noticeable changes from that of control pregnant women. Further studies are needed to verify their therapeutic and prophylatic roles against pre-eclamptic complication suring pregnancy. PMID- 12094144 TI - A case of Kearns-Sayre syndrome with autoimmune thyroiditis and possible Hashimoto encephalopathy. AB - The Kearns-Sayre syndrome (characterized by onset before 20, chronic ophthalmoplegia, pigmentary retinal degeneration and at least 1 of the following symptoms: ataxia, heart block and high protein content in the cerebrospinal fluid) is a severe variant of chronic progressive external ophthalmoplegia (CPEO) with frequent re-arrangements of the mitochondrial DNA (mtDNA). The aim of this paper is to report a sporadic paediatric case of Kearns-Sayre syndrome with mtDNA heteroplasmic deletion, absence of cytochrome c oxidase in many muscle fibers, autoimmune thyroiditis followed by depressive phobic disturbances, slowing EEG, hyperreflexia, tremor and visual hallucinations, in which the diagnosis of possible encephalopathy associated with autoimmune thyroid disease (Hashimoto encephalopathy) was made. We speculated that in this patient, predisposed by mitochondrial deletion, anti-thyroid antibodies may have interfered with mitochondrial cerebral function, causing Hashimoto encephalopathy and facilitating ophthalmoplegia. It seems important to study anti-thyroid antibodies in every case of Kearns-Sayre syndrome, specially if ophthalmoplegia is recent, even in order to the therapy. PMID- 12094146 TI - Treated Wilm's tumor in childhood as potential risk factor for second thyroid cancer. AB - The potential risk of a treatment-induced second neoplasia affecting the thyroid is well known after radiation therapy for several types of cancer, but few cases have been related to incidental irradiation for Wilms' tumor. We report a case of a papillary thyroid carcinoma discovered in a young patient 15 years after treatment of a Wilms' tumor. An 18-year-old man was referred to our Endocrinological Department for a single 3 cm nodule in the right lobe of the thyroid. His past medical history included at the age of 2 years surgical resection, chemotherapy (actinomycin-D and vincristine) and cesium radiation therapy to the right side for a Wilms' tumor in stage III: a total dose of 7700 rads was delivered to an area of 17 x 10 cm in the right flank. After fine-needle demonstration of a follicular thyroid lesion, the patient underwent right lobectomy, followed by total thyroidectomy for histologic diagnosis of a follicular variant papillary cancer. Residual thyroid tissue was ablated by iodine-131 administration (3700 MBq), but scanning after therapeutic iodine showed radioactive uptake in the left regional lymph nodes, with elevated serum thyroglobulin off therapy (830 ng/ml). Magnetic resonance imaging confirmed the presence of lymph node enlargements and bilateral neck dissection was performed, followed by radioiodine treatment (3700 MBq) and thyroxine suppressive therapy. After 3-year follow-up the patient is disease-free. Although few cases of thyroid cancer have been reported in the literature after irradiation for a Wilms' tumor during childhood, this association should be considered in the long-term follow up. PMID- 12094145 TI - Spontaneous regression and recurrence of primary low-grade B-cell gastric lymphoma on the gastric stump 15 and 20 years after gastroresection. AB - The recurrence of primary gastric lymphoma (PGL) on the gastric stump after gastroresection is rare. We describe the case of an 84-year-old man who had recurrences 15 and 20 years after a Billroth I gastrectomy. The concordance of the three gastric biopsies showing a low grade B-cell lymphocytic lymphoma of the mucosa-associated tissue, demonstrated the recurrence of the disease. The patient has serological evidence of Helicobacter pylori infection but the eradication therapy had no effect on the evolution of the disease. The case suggests that PGL is really a particular entity in the non-Hodgkin's lymphoma group, characterized by a long spontaneous natural history, with long lasting spontaneous remissions and recurrences. PMID- 12094147 TI - Dermatomyositis: diagnosis, evaluation and management. AB - Dermatomyositis (DM) is a condition primarily of the skin and muscles, but other systemic features may occur. DM in adults is associated with malignancy and thus a careful evaluation of each patient should be part of their initial and follow up assessments. Patients should also be evaluated for the presence of esophageal, pulmonary and cardiac disease. Corticosteroids, immunosuppressives, biologic agents and/or immune globulin are effective therapies for the myopathy of DM, whereas the skin disease is best managed with sunprotection, topical corticosteroids, antimalarials, methotrexate and/or immune globulin. The prognosis is good except for patients with malignancy, those with severe weakness and those with cardiopulmonary dysfunction. However, many patients are left with residual weakness even after control of the disease. PMID- 12094148 TI - The role of apoptosis in thyroid disease. AB - Increasing evidence suggests that apoptosis plays an important role in the pathogenesis of autoimmune and proliferative thyroid diseases, and that the apoptotic pathways involved are complex and highly regulated. Autoimmune thyroid diseases such as Hashimoto's thyroiditis and Graves' disease have been associated with differential expression of Fas and TRAIL receptor-mediated apoptosis. Thus, the thyroid cell destruction characteristic of autoimmune thyroiditis can be seen as the consequence of inappropriate expression of Fas or TRAIL death pathway molecules and down-regulation of the apoptosis controlling protein Bcl-2, which may be induced by cytokines released locally by infiltrating lymphocytes. In contrast, Graves' thyrocytes are protected from apoptotic death possibly by the anti-apoptotic action of thyrotrophin receptor antibodies or soluble Fas and/or the overexpression of Fas ligand which all create an anti-apoptotic potential for the thyroid cells and favor apoptosis of the infiltrating lymphocytes. On the other hand, an imbalance between thyroid cell proliferation and cell death may be crucial for goiter formation or cancer development and progression. In human thyroid goiter, Fas-mediated apoptosis is suppressed, leading to thyroid cell hyperplasia. Furthermore, malignant thyroid cells may escape immune attack by over expressing Fas ligand and inducing apoptosis in the invading immune cells. However, the exact mechanisms involved in the regulation of apoptosis in thyroid disease remain unclear. Further investigation is needed that may provide new strategies in the prevention and treatment of these diseases. PMID- 12094149 TI - [Ventricular arrhythmias. A potential risk associated with the use of non cardiovascular drugs prolonging the QT interval]. AB - The electrocardiogram shows a sequence of cardiac electrical events generated by individual electrical currents generated mainly by sodium (Na+), potassium (K+) and calcium (Ca++) ions transiting via specialized transport pathways, such as ion channels, inserted in the membranes of excitable cardiac cells. Na+ and Ca++, entering the cells, are messengers of activating, inward, depolarizing currents (INa, ICa), generating the QRS wave of the electrocardiogram, whereas K+, leaving the cells, carries outward repolarizing currents (e.g. Ito, IKr, IKs and IK1), generating the T wave of the electrocardiogram, which drives the cell to a rest condition. Therefore, a significantly prolonged repolarization process results in a prolonged QT interval which can initiate ectopic cardiac beats that may evolve into a potentially lethal ventricular tachyarrhythmia, called torsades de pointes. This implies that the measurement of QT interval, appropriately corrected (QTc) for heart rate values by applying available algorithms, is a current tool of diagnostic investigation to reveal abnormalities of the cardiac repolarisation process. Numerous non cardiovascular drugs in general clinical use (psychotropics, prokinetics, antimalarials, antibiotics, antihistamines, etc.) can prolong QT interval, often by a mechanism not necessary for their therapeutic effects, which is a reduction of the potassium current IKr conveyed by the HERG channel. The torsadogenic potential of these drugs can be facilitated by factors (female gender, bradycardia, electrolyte imbalances, cardiac diseases, simultaneous use of multiple drugs prolonging QT interval, etc.) known to have the propensity to reduce the redundant cardiac repolarization reserve, characterizing the healthy myocardium. Presently, drug candidates are routinely subjected to preclinical and clinical examination for cardiac safety, a property required by health authorities for any new medicine before allowing marketing authorization. Finally, before prescribing a medicinal product prolonging QT interval, a physician should carefully evaluate not only the disease he wants to treat but also the availability of equally effective, alternative drugs. The golden rule, to which such a prescription has always to abide, requires that the beneficial effects expected from a therapy should for each treated patient outweigh any possible adverse consequence, particularly when the latter one could be of lethal nature. PMID- 12094150 TI - The assessment of quality of life as a measure of gold salts treatment efficacy in rheumatoid arthritis. AB - BACKGROUND: In the evaluation of functional status of patients with rheumatoid arthritis (RA) the health related quality of life is currently considered important because of its approach to various components of life, such as social, psychological, and physical aspects. We used the Stanford Health Assessment Questionnaire (HAQ) to assess the improvement of functional status in patients with RA treated with gold salts. METHODS: In a prospective investigation 91 patients with RA in anatomical stage I, II, or III, 66 females and 25 males, with a mean age of 53.17 years, were evaluated during medical treatment in a 1-year follow-up. The treatment consisted in sodium aureothiosulphate, plus corticosteroids and NSAID. The assessments were done during ambulatory visits, at baseline and after 6 and 12 months of treatment, by HAQ as well as by other parameters such as Ritchie Index, visual analog scale (VAS), and morning stiffness. A group of 19 RA patients included by the same criteria and treated only by corticosteroids and NSAID was used as control for the first 6 months of the study. RESULTS: HAQ scores and other parameters were significantly lower (p=0.0001) at the 6th and 12th month measurements when compared with baseline. In the control group only a significant difference in the VAS score was detected. CONCLUSIONS: All the parameters measured in our study were useful in detecting clinical improvement in RA patients treated with sodium aureothiosulphate plus corticosteroids and NSAIDs, but the HAQ provides a more global assessment of the patient's status. PMID- 12094151 TI - [Prevalence and clinical features of fibromyalgia in systemic lupus erythematosus, systemic sclerosis and Sjogren's syndrome]. AB - BACKGROUND: We studied the prevalence of fibromyalgia in 3 different groups of patients affected respectively with systemic lupus erythematosus, systemic sclerosis (scleroderma) and primary Sjogren's syndrome. The typical fibromyalgia findings encountered in these diseases were examined. METHODS: We enrolled 250 consecutive outpatients: 100 with systemic lupus erythematosus, 50 with systemic sclerosis, 100 with primary Sjogren's syndrome and 2 control groups (30 healthy subjects and 75 patients with primary fibromyalgia). Fibromyalgia features were evaluated by algometry, VAS for pain, Mc Gill Pain Questionnaire and Fibromyalgia Impact Questionnaire. RESULTS: Fibromyalgia has been found in 1 case (1%) with systemic lupus erythematosus, 1 case with systemic sclerosis (2%), 22 cases (22%) with primary Sjogren's syndrome and in 1 (3.3%) of the healthy controls. The number of tender points was significantly higher (p<0.01) in the patients with Sjogren's syndrome in comparison with the other groups. Fibromyalgic findings were similar in the patients with primary fibromyalgia and Sjogren's syndrome with fibromyalgia, unless for both poor sleep and low algometric thresholds which were more frequently found in primary fibromyalgia (respectively p<0.001 and p=0.05). CONCLUSIONS: Our study suggests that fibromyalgia is relatively frequent in primary Sjogren's syndrome, while in systemic lupus and systemic sclerosis its prevalence is not different from that found in the healthy controls. Typical fibromyalgia findings, except algometric values, were similar between the cases with Sjogren's syndrome plus fibromyalgia and fibromyalgia alone. PMID- 12094152 TI - [Peptic ulcer hemorrhage in the elderly]. AB - BACKGROUND: The aim of this study was to investigate the efficacy of endoscopic injection therapy on the clinical outcome of elderly patients with peptic ulcer bleeding. METHODS: From January 1995 to December 1998, 738 patients with acute peptic ulcer bleeding were observed in the First Division of General Surgery, University of Verona: 359 aged <70 years and 379 =/>70 years. History, clinical and endoscopic findings and outcome were prospectively collected and analyzed comparing old (=/>70 yrs) and young (<70 yrs) patients. Ulcers with active or sign of recent bleeding were submitted to injection therapy using epinephrine and 1% polidocanol. RESULTS: Coexisting diseases were significantly more present in the elderly group except for liver cirrhosis that affected preferentially young patients (12.3 versus 4.0%; p<0.001). Endoscopic treatment was performed in a similar percentage between young and elderly patients (respectively 64.9 e 61.5%) and also the rebleeding rate (14.2 versus 13.2) and the mean duration of hospitalisation were not different. The overall mortality was 12.7% in the elderly group and 8.3% in the young group (p=0.04), whereas mortality after surgery was significantly higher in the young group (respectively 57.1 versus 8.3%; p=0.037). CONCLUSIONS: The clinical and endoscopic features and reebleeding rate were not different between elderly and young patients. Patients aged 70 years or older have a higher number of associated medical diseases except for liver cirrhosis conditions and a highest overall mortality whereas the risk of death after surgery is lower than in the younger group. PMID- 12094153 TI - [Merkel cell carcinoma of the skin in a patient with myasthenia gravis]. AB - Merkel cell carcinoma is a rare, aggressive neuroendocrine tumor of the skin commonly seen in the elderly on the head, neck and extremities, with a predisposition for local regional and distant spreading. A case of Merkel cell carcinoma occurred in a woman treated with immunosuppressive therapy for myasthenia gravis, is described and the possibility of a link between the immunosuppressive and/or oncogenic therapy and this tumor is suggested. PMID- 12094155 TI - An approach for trauma nurses to identify error-provoking factors. PMID- 12094154 TI - A time to change. PMID- 12094156 TI - Life in recovery: rebuilding from trauma. AB - Trauma can cause visible, often profound physical injuries for patients. The emotional and social drain that families and health care providers experience can also be life-altering. REBUILD is a program designed by a level 1 trauma center that incorporates former patients and their families in a support group to aid previous and new patients, their families, and care providers by sharing mutual experiences and learning from each other. Health care providers have found that participating in the program has been personally gratifying and professionally beneficial by preventing burn-out. PMID- 12094157 TI - Does caring for displaced specialty unit patients affect the critical care nurse's perceptions of ability and job satisfaction? AB - Trauma centers are challenged to share beds with a larger hospital population of critical care patients. Often, this means that patients may be shifted between units when beds are not immediately available in the specialty unit that fits their diagnosis. They are admitted to the first intensive care unit bed that becomes available. This practice results in patients with special care needs being cared for by nursing staff who do not perceive themselves as trained to provide those needs. This practice is referred to as displaced specialty unit (DSU) admission. A review of 2-year data from one large trauma center revealed a total of 1072 DSU patients, of whom 50% were medical patients. A questionnaire given to intensive care unit nurses found that caring for DSU patients did affect their perceptions of their ability to care for such patients and affected their sense of job satisfaction. Strategies to improve nurses' comfort level and competency in treating diverse critical care patients were recommended and implemented. PMID- 12094158 TI - Child maltreatment awareness for prehospital providers. AB - Child abuse continues to go undetected by hospital-based health care providers because clues may exist exclusively in the child's home. Emergency personnel are in the unique position of being able to assess the home environment. An educational intervention was developed to increase the prehospital providers' awareness of how to assess, report, and document suspected child maltreatment based on findings in the home. PMID- 12094160 TI - Traffic safety: ambitious targets and initiatives in Canada and around the world. PMID- 12094159 TI - Should families be allowed in trauma resuscitations? PMID- 12094161 TI - Better health care with quality medical devices: FDA on the cutting edge of device technology. PMID- 12094163 TI - Worldwide injection safety, misuse and overuse. PMID- 12094164 TI - Low-level exercise echocardiography identifies contractile reserve in patients with a recent myocardial infarction: comparison with dobutamine stress echocardiography. AB - Contractile reserve (CR) during dobutamine stress echocardiography (DSE) identifies patients with coronary artery disease whose left ventricular function and prognosis may improve after revascularization. To test the hypothesis that the early stages of upright bicycle-exercise echocardiography (LLEx) result in CR comparable with DSE, 30 patients 1 week after myocardial infarction underwent multistage LLEx and DSE. At low and peak LLEx, heart rate and systolic blood pressure were higher than DSE. Identification of CR by LLEx was concordant with DSE when analyzed by infarct zone or by individual patient. We conclude that CR can be demonstrated during LLEx that correlates with DSE in patients with a recent MI. Low-level exercise is a promising alternative to DSE for identifying contractile reserve after MI. PMID- 12094165 TI - The role of quantitative myocardial contrast echocardiography in the study of coronary microcirculation in athlete's heart. AB - Quantitative myocardial contrast echocardiography was performed with harmonic power Doppler analysis using the background subtraction and Levovist (Schering AG, Berlin, Germany) as contrast agent in a triggered modality. Quantitative analysis of echocontrast was performed offline with PC software, obtaining the transit curves of microbubbles through the coronary capillary system. Coronary microcirculation in athletes showed a behavior substantially comparable with control participants, although at a higher level. Training determines a physiologic left ventricular hypertrophy that counterbalances the dilatation in the left ventricular chambers because of the higher blood volume in athletes compared with control participants. Angiogenesis and several functional adaptations (relaxation of small coronary arteries, increased production of nitric oxide by the coronary endothelium, or both), represent the potential mechanisms that allow an optimal distribution of oxygen and of substrates to the hypertrophied myocardium of the athletes. PMID- 12094166 TI - Evaluation of a new ultrasound contrast agent (AI-700) using two-dimensional and three-dimensional imaging during acute ischemia. AB - BACKGROUND: A new intravenous contrast agent, AI-700, was evaluated to determine whether a bolus injection could be used to detect myocardial perfusion abnormalities during acute ischemia by using 2-dimensional (2D) and 3-dimensional (3D) myocardial contrast echocardiography. METHODS: 2D MCE was performed in 14 closed-chest dogs during coronary occlusion by using both continuous and triggered gray scale harmonic imaging and triggered power Doppler imaging. 3D MCE (open-chest) and nuclear perfusion imaging were performed in 10 of the 14 dogs. Postmortem triphenyl tetrazolium chloride (TTC) staining was performed to verify infarction. RESULTS: Thirteen of the 14 dogs had infarct by TTC; all 10 that had nuclear imaging showed a perfusion defect. Of the 13 dogs that had infarction, perfusion defects were detected in all (13 of 13) by gray scale harmonic imaging (sensitivity = 100%), and in 11 of 13 by power Doppler imaging (sensitivity = 85%). All 10 dogs that had nuclear imaging showed perfusion defects by gray scale harmonic imaging (sensitivity = 100%) and 8 of 10 by power Doppler imaging (sensitivity = 80%). The perfusion defect size, derived from 3D imaging (25% +/- 12%) correlated well with that from nuclear imaging (24% +/- 12%) (y = 0.9x + 3.8, r = 0.96, mean difference = 1.3% +/- 2.6%). The perfusion defect mass by 3D (22 +/- 14 g) also correlated well with the infarct mass by TTC staining (24 +/- 16 g) (y = 0.8x + 2.9, r = 0.89, P <.001, mean difference = -2.8 +/- 7.6 g). CONCLUSION: After a single bolus of AI-700, both 2D and 3D MCE could accurately detect perfusion defects representing the area at risk of infarction during acute ischemia compared with nuclear imaging and predicted the size of infarction as verified by TTC staining. PMID- 12094168 TI - Better prognosis of elderly patients with infectious endocarditis in the era of routine echocardiography and nonrestrictive indications for valve surgery. AB - OBJECTIVE: It has been reported that endocarditis in the elderly may have a poor outcome. Our aim was to assess the different features and prognosis, if any, in the present time. METHODS: Of 103 patients with proven endocarditis, 31 were 65 years or older and 72 were younger than 65 years. Degenerative heart disease was seen more frequently in the elderly (22.5% vs 2.7%, P =.003). Drug abuse and immunodeficiency virus infection were more common in the younger group, as was tricuspid endocarditis (26.3% vs 0%, P <.001). At clinical presentation cardiac failure (41.9 vs 19.4%, P =.02) and leukocytosis (61.2% vs 40.2%, P =.049) were seen more frequently in the elderly. RESULTS: Despite other similar clinical features, it took longer to diagnose older patients (7.2 +/- 6.2 vs 3.2 +/- 3.5 days, P <.001). Enterococcus infected the aged more often (32.2% vs 13.1%, P =.001). During hospitalization, heart failure and embolization tended to be more common in the elderly and the younger group, respectively. There were no significant differences in the incidence of anatomic complications, the need for operation, and overall mortality. CONCLUSION: Although a worse prognosis has been reported in elderly patients with infective endocarditis, the early use of transesophageal echocardiographic examinations and equal therapeutic options provides a similar outcome when compared with younger subjects. PMID- 12094167 TI - Enhanced method for predicting left ventricular reverse remodeling after surgical repair of aortic regurgitation: application of ultrasonic tissue characterization. AB - To predict left ventricular (LV) reverse remodeling after surgical repair of aortic regurgitation, we examined 30 patients with aortic regurgitation accompanying LV dilatation by myocardial tissue characterization with integrated backscatter method. Before and after operation, the magnitude of cyclic variation of integrated backscatter (CVIB) was obtained from anterior septum and posterior wall, and averaged value was calculated in each patient. Before operation, LV end diastolic dimension, fractional shortening, and LV end-diastolic pressure were not significantly different between the patients with (group GR) and without (group PR) decreased LV end-diastolic dimension after operation. Under these conditions, CVIB, which was 9.6 +/- 1.0 dB from healthy volunteers, was significantly greater in group GR, 5.7 +/- 1.4 dB, than that in group PR, 3.8 +/- 0.8 dB (P =.0003). The patients with CVIB >/= 4 before operation were expected to have reverse remodeling after operation with a sensitivity of 79%, a specificity of 82%. These data indicate that preoperative CVIB from the left ventricle provides pivotal information for predicting reverse remodeling after operation for aortic regurgitation in addition to the conventional echocardiographic parameters. PMID- 12094169 TI - Use of pulse wave and color flow Doppler echocardiography in mouse models of human disease. AB - The noninvasive assessment of cardiovascular physiology in mice is challenging because of their small size and extremely rapid heart rates. In this study, we sought to determine the feasibility and utility of pulse wave (PW) and color flow Doppler imaging techniques when applied to mouse models of cardiac remodeling. We performed transverse aortic banding, induced aortic insufficiency (AI), or sham procedures in wild-type mice. Animals were anesthetized and transthoracic echocardiography was performed, including PW and color flow Doppler imaging. In aortic banded mice, color flow Doppler imaging was used to identify flow around the aortic arch, and PW measurements were performed distal to the constriction. A high-velocity jet across the aortic constriction was detected in all 4 banded animals. Both modalities were applied to AI mice in which AI was detected in all 5 mice but in none of the 5 shams. Pulse wave and color flow Doppler imaging were also used to screen senescent mice for valve abnormalities; AI was detected in 3 of 20 and mitral regurgitation in 2 of 20. We demonstrate here that PW and color flow Doppler imaging techniques are useful in the evaluation of mouse models of cardiac remodeling. In addition, this study indicates that these modalities may be potentially useful to screen transgenic mice for valve abnormalities. PMID- 12094170 TI - The mechanism of emergence and clinical significance of apically directed intraventricular flow during isovolumic relaxation. AB - The mechanism of emergence and the clinical significance of apically directed intraventricular flow during isovolumic relaxation were investigated. The relation between the spatial distribution of the flow and left ventricular (LV) apical wall motion abnormality, as well as LV performance, was studied in 97 patients who underwent cardiac catheterization for evaluation of chest pain. According to the distribution of the flow, the patients were classified into the following 3 groups: flow observed in the whole area between the tip of the papillary muscle and the apex (spread flow) (n = 38), flow observed in the same area that did not fill the whole area (localized flow) (n = 15), and no apparent flow observed in the area (without flow) (n = 44). An absence of flow disclosed apical asynergy with a sensitivity of 97% and specificity of 87%. The time constant of LV relaxation was significantly shorter in patients with spread flow than in those without flow. A significant difference was also observed in end systolic volume index (18.8 +/- 6.8 vs 30.9 +/- 7.7 vs 42.3 +/- 20.2 mL/m(2), spread flow < localized flow < without flow, P <.05) among the 3 groups. The propagation velocity of LV early diastolic filling flow was significantly greater in patients with spread flow (47.0 +/- 8.3 cm/s) than in those with localized flow (30.7 +/- 7.8 cm/s) or without flow (28.6 +/- 7.8 cm/s) (P <.001). These findings indicate that the greater magnitude of LV elastic recoil and the faster LV relaxation in patients without LV apical asynergy produce apically directed intraventricular flow during isovolumic relaxation, enhancing the speed of LV early diastolic filling. Apically directed intraventricular flow during isovolumic relaxation may play an important role as a mediator of better LV systolic performance and LV relaxation to LV early diastolic filling. Absence of apically directed intraventricular flow during isovolumic relaxation is a manifestation of LV apical asynergy and global LV dysfunction from end systole to early diastole. PMID- 12094171 TI - Can changes in systolic longitudinal deformation quantify regional myocardial function after an acute infarction? An ultrasonic strain rate and strain study. AB - OBJECTIVES: The aim of this study was to evaluate the additional value of ultrasonic strain rate and strain to myocardial velocity in the identification and quantification of regional asynergy after an acute myocardial infarction (MI). METHODS: Forty patients (59 +/- 13 years) were investigated 3 +/- 2 days after a first infarction and compared with 14 age-matched controls with normally contracting segments (group A, n = 146). Longitudinal myocardial velocities, strain rate (SR) and strain (epsilon) were postprocessed from basal, mid, and apical segments interrogated using apical views. In a subset of patients with coronary angiograms (n = 24), myocardial segments were divided into 3 groups: normally contracting segments supplied by a normal coronary artery (group B1), normally contracting segments supplied by a diseased coronary artery (group B2), and segments with abnormal motion (group B3). Velocities were decreased in patients with myocardial infarction (MI) (P <.05 vs controls) but failed to accurately differentiate normally from abnormally contracting segments. At the opposite end, systolic SR and epsilon decreased significantly with segmental asynergy severity and could identify infarct-involved segments (group B3) with a sensitivity/specificity of 85% (systolic SR and epsilon cutoff values of -0.8 s( 1) and -13%, respectively). CONCLUSION: Strain rate and strain can better assess segmental dysfunction severity than myocardial velocities alone after an acute MI. PMID- 12094172 TI - Errors as a result of metal in the near environment when using an electromagnetic locator with freehand three-dimensional echocardiography. AB - BACKGROUND: Quantitative ventriculography by freehand 3-dimensional (3D) echocardiography with an acoustic spatial locator has been proven to provide highly accurate reproducible measurements of left ventricular volume, mass, and function. It has been shown to be 2 to 3 times better than conventional 2 dimensional echocardiographic techniques. Although accurate, the acoustic spatial locator uses a spark gap to generate hypersound for locating and is somewhat bulky. The Bird direct current electromagnetic locator (Ascension Technology Corp, Burlington, Vt) is a notable alternative locator for the freehand 3D system because it is small and easily portable. However, conductive metals in the near environment may adversely affect electromagnetic locator accuracy. To determine the feasibility of using the electromagnetic locator in a freehand 3D echocardiographic system in the conventional hospital environment, a series of experiments was carried out assessing the accuracy of such a system under various conditions of exposure to conductive metal. METHODS: Using tissue equivalent ellipsoid phantoms of known volumes, we compared volume measurement accuracy of the freehand 3D echocardiographic system equipped with the standard Bird or miniBird electromagnetic locator systems with our freehand acoustic spatial locator 3D echocardiographic system in 3 experiments: (experiment 1) no metal within 30 in (76.2 cm) of the phantoms and electromagnetic locator; (experiment 2) phantoms placed on a standard metal hospital stretcher with conductive metal less than 10 in (25.4 cm) from the phantoms and electromagnetic locator and with the echocardiographic machine greater than 30 in (76.2 cm) from the electromagnetic locator; and (experiment 3) phantoms placed on the same stretcher with conductive metal less than 10 in (25.4 cm) from the phantoms and electromagnetic locator and with the echocardiographic machine in its usual position approximately 10 in (25.4 cm) from the electromagnetic locator. RESULTS: For experiment 1 there was no significant volume error (<1%) by any system; no significant difference among the 3 locator systems (acoustic, Bird, or miniBird). For experiment 2 there was significant volume underestimation error by both electromagnetic locator systems (-10.9%, P <.05). For experiment 3 there was significant and greater volume underestimation error by both electromagnetic locator systems (-14.7%, P <.05) in close proximity to the echocardiographic machine. Interobserver variability was 5.1%. CONCLUSION: For quantitative ventriculography by a freehand 3D echocardiographic system, electromagnetic locator systems should not be used if conductive metal is in the near environment (<30 in [76.2 cm] from the locator). Accurate quantitative ventriculography may be performed with an electromagnetic locator system if the near environment is free of conductive metals. PMID- 12094173 TI - Congenital right coronary artery aneurysm causing myocardial infarction, pseudoaneurysm formation, and right atrial compression. AB - We present a case of a young woman with symptoms of acute myocardial infarction and found to have a nonatherosclerotic coronary aneurysm of the right coronary artery, which ruptured and formed a pseudoaneurysm. The pseudoaneurysm was sufficiently large that it mimicked an atrial mass on diagnostic imaging modalities. With the attendant risk of further rupture and compression of the heart chambers, the patient underwent successful resection of the aneurysm and pseudoaneurysm with single vessel bypass surgery. PMID- 12094174 TI - Transthoracic echocardiographic visualization of coronary artery blood flow and assessment of coronary flow reserve in the right coronary artery: a first report of 3 patients. AB - Assessment of coronary flow reserve (CFR) has proven to be an important diagnostic tool providing useful clinical and physiologic information about coronary artery function. In several studies, the transthoracic echocardiographic assessment of CFR, defined as a ratio of hyperemic to basal coronary flow velocity, was validated in the left anterior descending artery. But so far, the visualization of coronary flow and the measurement of CFR were limited to the mid and distal portion of the left anterior descending artery. Introduction of a modified 2-chamber view enables the recording of coronary blood flow and the assessment of CFR in the posterior descending branch of the right coronary artery in selected patients. This report of 3 cases describes for the first time a method to visualize and measure coronary blood flow in the distal right coronary artery by precordial Doppler echocardiography. PMID- 12094175 TI - False aneurysm of the mitral-aortic intervalvular fibrosa after uncomplicated aortic valve replacement. AB - False aneurysms of the mitral-aortic intervalvular fibrosa are rare and usually complicate aortic valve endocarditis. We report a case of a false aneurysm of the mitral-aortic intervalvular fibrosa after recent bioprosthetic aortic valve replacement in the absence of endocarditis. PMID- 12094176 TI - Mitral valve thrombus mimicking a primary tumor in the antiphospholipid syndrome. AB - Cardiac valvular abnormalities detected by echocardiography are relatively common in patients with the primary antiphospholipid syndrome. Valvular thickening with small vegetations are typical, but reports of lesion histology are rare. We report the case of a 50-year-old man presenting with thromboembolic phenomena who had a large (>2 cm), mobile, pedunculated, mass attached to the mitral valve that had the echocardiographic appearance of a primary valve tumor. Following surgical removal, histopathologic examination demonstrated pure fibrin thrombus and serological testing confirmed the antiphospholipid syndrome. This case highlights an unusual echocardiographic appearance of intracardiac thrombus in this syndrome. PMID- 12094178 TI - Prominent crista terminalis appearing as a right atrial mass on transthoracic echocardiogram. AB - The crista terminalis is a fibromuscular ridge formed by the junction of the sinus venosus and primitive right atrium. In these case reports, transthoracic echocardiography suggested the presence of a right atrial mass. However, subsequent transesophageal echocardiography revealed that the "right atrial mass" was actually a prominent crista terminalis. An understanding of the anatomy and the echocardiographic appearance of a prominent crista terminalis will minimize the misdiagnosis of this structure. PMID- 12094177 TI - Visceroatrial situs solitus with atrioventricular alignment discordance double outlet right ventricle and superoinferior ventricles: fetal and neonatal echocardiographic findings. AB - Visceroatrial situs, ventricular morphology, and atrioventricular and ventriculoarterial alignments can reliably be assessed with fetal echocardiography. We herein describe, to our knowledge, the first reported fetal echocardiographic diagnosis of a fetus with visceroatrial situs solitus, atrioventricular alignment discordance, double outlet right ventricle, and superoinferior ventricles of a 22-year-old gravida III para II woman with a previous history of intrauterine fetal death. She had been evaluated at another center and was referred to our unit because of the suspicion about her diagnosis and underwent fetal ultrasonographic evaluation at 39 weeks gestation. Neonatal echocardiographic evaluation and autopsy findings confirmed the diagnosis. Accordingly there was (solitus, L-loop ventricle, and anterior aorta) double outlet right ventricle, superoinferior ventricles with diminutive right ventricle, noncommitted trabecular ventricular septal defect, and pulmonary hypertension. PMID- 12094179 TI - Cardiac tuberculosis with multiple intracardiac masses: a case report. AB - In this report, clinical, echocardiographic, and pathologic findings of a patient with multiple masses caused by tuberculosis both in the left and right side of the heart are presented. After antituberculosis treatment some of the masses disappeared and some became smaller. Although an intracardiac mass caused by tuberculosis is very rare, it should be considered in the list of masses detected by echocardiography. PMID- 12094180 TI - The state of exercise training: a need for action. PMID- 12094181 TI - Color me variable: the irreproducible nature of color flow Doppler in mitral and aortic regurgitation. PMID- 12094182 TI - Selective L-type, T-type, and nonspecific calcium-channel blockers for stable angina pectoris. PMID- 12094183 TI - Transient atrial mechanical dysfunction (stunning) after cardioversion of atrial fibrillation and flutter. AB - BACKGROUND: Conversion of atrial fibrillation (AFib) and flutter (AFlt) to sinus rhythm results in a transient mechanical dysfunction of atria (atrial stunning). Methods used as a means of assessing atrial stunning, atrial stunning after conversion of atrial fibrillation/flutter, and the cause, mechanisms, determinants of the extent, and drugs affecting atrial stunning were examined. METHODS: Studies on the subject, identified through a comprehensive literature search, were thoroughly evaluated. RESULTS AND CONCLUSIONS: Left atrial (LA) stunning has been reported with all modes of conversion of AFib/AFlt to sinus rhythm. The incidence of LA stunning is 38% to 80%. Spontaneous echocardiographic contrast, LA appendage (LAA) flow velocities and emptying fraction, transmitral inflow velocity of atrial wave (A-wave), time-velocity integral of A-wave, and atrial filling fraction have been used as means of assessing LA stunning. The data on right atrial (RA) stunning are limited, but parallel findings have been reported in the right atrium. Atrial stunning does not develop after the unsuccessful attempts of cardioversion or on delivery of electric current to the heart without AFib/AFlt, and it is a function of the underlying AFib/AFlt manifesting at the restoration of sinus rhythm. Tachycardia-induced atrial myopathy and chronic atrial hibernation are suggested mechanisms. Duration of preceding AFib/AFlt, atrial size, and underlying heart disease are determinants of the extent of atrial stunning. Verapamil, dofetilide, and acetylstrophenathidine have been shown to attenuate or protect from atrial stunning in animal or small human studies. A comprehensive knowledge of atrial stunning would be helpful in selecting the patients for, and the duration of, anticoagulation therapy after cardioversion. PMID- 12094184 TI - Effects of exercise training in patients with heart failure: the Exercise Rehabilitation Trial (EXERT). AB - BACKGROUND: The purpose of this study was to examine the effects of exercise training on functional capacity in patients with heart failure. METHODS: One hundred eighty-one patients in New York Heart Association class I to III, with ejection fraction <40% and 6-minute walk distance <500 meters, were recruited into a randomized, controlled, single-blind trial comparing 3 months of supervised training, then 9 months of home-based training with usual care. RESULTS: There was a significant increase in 6-minute walk distance at 3 and 12 months but no between-group differences. Incremental peak oxygen uptake increased in the exercise group compared with the control group at 3 months (0.104 +/- 0.026 L/min vs 0.025 +/- 0.023 L/min; P =.026) and 12 months (0.154 +/- 0.074 L/min vs 0.024 +/- 0.027 L/min; P =.081). Compared with the control group, significant increases were observed in the exercise group for arm and leg strength. No significant changes were observed in cardiac function or quality of life. Adherence to exercise was good during supervised training but reduced during home-based training. CONCLUSIONS: Exercise training improves peak oxygen uptake and strength during supervised training. Over the final 9 months of the study, there was little further improvement, suggesting that some supervision is required for these patients. There were no adverse effects on cardiac function or clinical events. PMID- 12094185 TI - Diuretic resistance predicts mortality in patients with advanced heart failure. AB - BACKGROUND: In patients with chronic heart failure (CHF), diuretic requirements increase as the disease progresses. Because diuretic resistance can be overcome with escalating doses, the evaluation of CHF severity and prognosis may be incomplete without considering the intensity of therapy. METHODS: The prognostic importance of diuretic resistance (as evidenced by a high-dose requirement) was retrospectively evaluated in 1153 patients with advanced CHF who were enrolled in the Prospective Randomized Amlodipine Survival Evaluation (PRAISE). The relation of loop diuretic and angiotensin-converting enzyme inhibitor doses (defined by their median values) and other baseline characteristics to total and cause specific mortality was determined by proportion hazards regression. RESULTS: High diuretic doses were independently associated with mortality, sudden death, and pump failure death (adjusted hazard ratios [HRs] 1.37 [P =.004], 1.39 [P =.042], and 1.51 [P =.034], respectively). Use of metolazone was an independent predictor of total mortality (adjusted HR = 1.37, P =.016) but not of cause-specific mortality. Low angiotensin-converting enzyme inhibitor dose was an independent predictor of pump failure death (adjusted HR = 2.21, P =.0005). Unadjusted mortality risks of congestion and its treatment were additive and comparable to those of established risk factors. CONCLUSIONS: The independent association of high diuretic doses with mortality suggests that diuretic resistance should be considered an indicator of prognosis in patients with chronic CHF. These retrospective observations do not establish harm or rule out a long-term benefit of diuretics in CHF, because selection bias may entirely explain the relation of prescribed therapy to death. PMID- 12094186 TI - Peripheral endothelium-dependent flow-mediated vasodilatation is associated with left ventricular mass in older persons with hypertension. AB - BACKGROUND: Increased left ventricular (LV) mass is associated with greater cardiovascular disease risk. Recent studies have also shown an association of increased LV mass with attenuated endothelium-dependent coronary flow reserve. Less is known about the association between LV mass and endothelium-dependent flow-mediated dilatation (FMD) in peripheral arteries, a noninvasive measure of endothelial function. METHODS: Sixty-two subjects with untreated mild hypertension, aged 55 to 75 years and otherwise healthy, were examined. Resting blood pressure was obtained by the average of 4 to 5 visits, each at least 1 week apart. LV mass was determined from magnetic resonance imaging and was indexed by body surface area, height and height(2.7). Body composition was assessed with dual energy x-ray absorptiometry. FMD was measured as the percent change of brachial artery diameter during reactive hyperemia by use of high-resolution ultrasound. RESULTS: Median LV mass index was 63 g/m2 (interquartile range, 58 73). In bivariate analysis, LV mass was correlated to lean body mass (r = 0.63, P <.001), diastolic blood pressure (r = 0.35, P <.01), and FMD (r = -0.27, P <.05). In multivariate analysis, 44% of the variance in log-LV mass was explained by lean body mass. An additional 6% of the variance was explained by FMD (P <.05). For each 1% point decrease in FMD, LV mass increased by 1.1%. CONCLUSIONS: In addition to the expected influences of body size, impairment of brachial artery FMD was independently related to LV mass in elderly subjects with mild hypertension who did not yet have LV hypertrophy. Whether mild hypertension is the common mechanism linking LV mass and endothelial function has yet to be determined. PMID- 12094187 TI - The cardiac component of cardiac cachexia. AB - BACKGROUND: Recent evidence suggests the importance of noncardiac mechanisms in the genesis of the syndrome of cardiac cachexia. This raises the question of the relative role of the heart itself in this syndrome. This study sought to assess the cardiac dimensions, mass, and function and changes in these parameters over time in patients with chronic heart failure with and without cachexia. METHODS: Doppler echocardiography was performed in 28 patients with nonedematous weight loss (>7.5% over a period of >6 months) compared with 56 matched patients without weight loss in a ratio of 1:2 (age 71 +/- 13 vs 67 +/- 8 years, P =.07; New York Heart Association class 2.9 +/- 0.7 vs 2.6 +/- 0.6, P =.08). In 18 cachectic and 35 noncachectic patients with previous echocardiographic recordings, we analyzed the changes in left ventricular (LV) dimensions and mass over time. RESULTS: Cardiac dimensions including LV diastolic (69 +/- 9 mm vs 67 +/- 13 mm) and systolic cavity diameter (58 +/- 11 mm vs 55 +/- 15 mm), LV mass (480 +/- 180 g vs 495 +/- 190 g), and LV systolic and diastolic function including fractional shortening (16% +/- 10% vs 18% +/- 10%), isovolumic relaxation time (29 +/- 22 ms vs 36 +/- 27 ms), and E/A ratio (2.7 +/- 1.6 vs 3.3 +/- 2.9) did not differ between cachectic and noncachectic patients (all P >.1). By analyzing changes in LV mass over time, we found an increase (>20%) in 2 (11%) cachectic and 14 (40%) noncachectic patients and a decrease in LV mass (>20%) in 9 (50%) cachectic and 8 (23%) noncachectic patients (chi2 test, P <.05). CONCLUSIONS: Although no specific cardiac abnormality could be detected echocardiographically in cachectic patients compared with patients with noncachectic chronic heart failure in a cross-sectional study, over time a significant loss of LV mass (>20%) occurs more frequently in patients with cardiac cachexia. PMID- 12094188 TI - Perceptions of chest pain differ by race. AB - BACKGROUND: African American patients are less likely to receive thrombolytic therapy and coronary revascularization than are white patients. Delay and clinical presentation may be keys to understanding differences in care. OBJECTIVE: To determine how symptom recognition and perception influence clinical presentation as a function of race, we characterized symptoms and care-seeking behavior in African American and white patients seen in the ED with chest pain. METHODS: The prospective study was conducted from April 1999 to September 1999 among patients who were seen in the ED and were admitted or observed in the ED Chest Pain Unit (n = 215). Interviews were conducted within 48 hours with a structured set of questions. RESULTS: Thirty-one percent of white patients and 8.9% of African American patients were admitted with a diagnosis of acute myocardial infarction (P =.001). African American patients were as likely as white patients to report "typical" objective symptoms but were more likely to attribute their symptoms to a gastrointestinal source rather than a cardiac source (P =.05). Of those patients with the final diagnosis of myocardial infarction (n = 45), 61% of African American patients attributed symptoms to a gastrointestinal source and 11% to a cardiac source, versus 26% and 33%, respectively, for white patients. The median prehospital delay for African American patients was 263 minutes (interquartile range, 120 to 756 minutes), similar to the 247 minutes for white patients (interquartile range, 101 to 825 minutes, P =.72), despite African American patients (80%) being more likely than white patients (66%) to perceive their symptoms as severe/life-threatening at onset (P =.05). CONCLUSION: Racial differences in symptom perception exist. Although the proportion of objectively defined typical symptoms were similar, self-attribution was more often noncardiac in African American patients than in white patients. Self-attribution, in addition to objective clinical findings, is likely to influence caregiver diagnostic approaches and therefore therapeutic approaches, and merits further study. PMID- 12094190 TI - Effect of beta2-adrenergic receptor functioning and increased norepinephrine on the hypercoagulable state with mental stress. AB - BACKGROUND: Procoagulant stress responses may contribute to atherosclerosis development and acute coronary thrombosis. In the present study, we examined the role of beta2-adrenergic receptor function and plasma catecholamines in the stress-induced increase in the 2 hypercoagulability markers thrombin-antithrombin III (TAT) complex and fibrin D-dimer (DD). METHODS: Lymphocyte beta2 adrenoreceptor sensitivity and density were assessed at rest, and plasma levels of TAT, DD, epinephrine, and norepinephrine were measured at rest and in response to a standardized mental stress task in 19 normotensive and mildly hypertensive nonmedicated subjects (mean age 38 years, age range 29 to 48 years). RESULTS: The stressor elicited a significant increase in TAT (P =.024), DD (P =.026), and norepinephrine (P =.005). Resting beta2-adrenoreceptor sensitivity (isoproterenol stimulated cyclic adenosine monophosphate production) plus the norepinephrine change scores (stress minus rest) accounted for 59% of the variance in the absolute TAT increase in response to stress (P =.001). Hypertension status and demographic variables such as sex did not influence the results. CONCLUSIONS: Acute mental stress may trigger a hypercoagulable state evidenced by increased thrombin activity and increased fibrin turnover. Beta2-adrenergic receptor sensitivity and plasma catecholamine activity may mediate the procoagulant response to acute stressors. These mechanisms may help explain the adverse impact of mental stress on the cardiovascular system. PMID- 12094189 TI - Randomized comparison of T-type versus L-type calcium-channel blockade on exercise duration in stable angina: results of the Posicor Reduction of Ischemia During Exercise (PRIDE) trial. AB - BACKGROUND: Mibefradil is a T-type calcium-channel antagonist and arterial vasodilator with negative chronotropic effects. It is not known if T-type calcium channel blockade is superior to L-type calcium-channel blockade in patients with stable angina pectoris. METHODS: A multicenter, randomized, double-blind trial was conducted in patients with documented coronary disease and stable angina to compare a 360 mg dose of diltiazem CD with 100 mg dose of mibefradil. The primary end point was change in time to symptom-limited exercise termination from baseline to 8 weeks. Secondary efficacy parameters included time to onset of persistent ST-segment depression, time to awareness of angina, and change in exercise duration from baseline to 2 and 4 weeks of treatment. RESULTS: A total of 121 patients were randomized to mibefradil and 113 to diltiazem CD. At 8 weeks, the increase in exercise duration was 24.5 seconds greater in the mibefradil group (P =.017; 95% CI 4.4-44.7 seconds). At 8 weeks, time to development of > or =1 mm ST-segment depression was greater by 45.3 seconds (P =.0025; 95% CI 16.2-74.5) with mibefradil, but time to development of angina was not significantly different. CONCLUSION: T-type calcium-channel antagonism with mibefradil improved treadmill exercise parameters compared with diltiazem in patients with chronic stable angina. Further investigation and development of antagonists of T-type calcium channels with fewer adverse drug interactions is warranted and may be promising in the management of ischemic heart disease. PMID- 12094191 TI - Heparin dosing and outcome in acute coronary syndromes: the GUSTO-IIb experience. Global Use of Strategies to Open Occluded Coronary Arteries. AB - BACKGROUND: This study analyzed relationships among heparin dosage, patient characteristics, and 30-day outcome because optimal unfractionated-heparin dosing in acute coronary syndromes remains uncertain. METHODS: Patients (n = 5335) randomized to heparin therapy in the Global Use of Strategies to Open Occluded Coronary Arteries (GUSTO) IIb trial were studied. The heparin dose was adjusted to a target activated partial thromboplastin time (aPTT) and normalized for the patient's weight. Mortality and cardiac (re)infarction within 30 days and their association with patient characteristics and heparin dosing were evaluated. RESULTS: The lowest mortality rate appeared with a heparin dose of approximately 14 U/kg/h or an aPTT of approximately 70 seconds. Heparin dosing was a significant predictor of outcome after adjusting for presenting coronary syndrome; a trend remained after adjusting for other baseline differences. This association was lost when adjusted for the aPTT result. Patients who died early appeared to have lower heparin dosing than those with later mortality (P =.012). Heparin "resistance" with relatively high heparin dosages and low aPTT values did not increase the risk for adverse outcome. CONCLUSIONS: There is a defined, dose associated benefit of unfractionated heparin in acute coronary syndromes similar to that seen previously in thrombolytic-treated infarctions. Heparin therapy is complicated by its complex biologic interactions and relatively crude measures of its effect. Better measures of heparin effectiveness and strategies need to be developed with either better antithrombin agents or adjunctive therapies such as antiplatelet regimens to treat patients who require benefits beyond that supplied by unfractionated heparin. PMID- 12094192 TI - Predicting outcome after thrombolysis in acute myocardial infarction according to ST-segment resolution at 90 minutes: a substudy of the GUSTO-III trial. Global Use of Strategies To Open occluded coronary arteries. AB - BACKGROUND: Resolution of ST-segment elevation after thrombolysis for acute myocardial infarction has been shown to have prognostic significance 3 hours (180 minutes) after the initiation of therapy. Whether prognostically useful information can be achieved as early as 90 minutes after thrombolysis is unknown. METHODS: An electrocardiographic substudy of 2352 patients from the Global Use of Strategies To Open occluded coronary arteries (GUSTO-III) trial was undertaken to compare outcomes according to ST-segment resolution at 90 minutes versus 180 minutes after administration of thrombolytic therapy. RESULTS: Of 2352 patients in the substudy, 2241 had a baseline and 90-minute electrocardiogram, and 2218 had a baseline and 180-minute ECG. Complete ST-segment resolution occurred in 44.2% of patients at 90 minutes and 56.5% of patients at 180 minutes. ST-segment resolution at both 90 and 180 minutes was associated with lower 30-day and 1-year mortality. Multivariate analysis revealed ST-segment resolution at 90 minutes to be an equally strong predictor of 30-day mortality as resolution at 180 minutes. Patients who were at particularly high risk for mortality were those aged >70 years, those who presented with Killip class >1, and those with anterior infarctions. CONCLUSIONS: The presence of ST-segment resolution on standard 12 lead electrocardiographic monitoring 90 minutes after thrombolysis is a useful independent predictor of mortality at 30 days and 1 year. The potential for obtaining prognostic results as early as 90 minutes after thrombolysis sets a new precedent for optimum electrocardiographic monitoring times in these patients. PMID- 12094193 TI - Prognosis after acute myocardial infarction continues to improve in the reperfusion era in the community of Goteborg. AB - BACKGROUND: The objective of this study was to compare the prognosis of nonselected patients who had an acute myocardial infarction (AMI) during 2 time periods in the thrombolytic era and to describe coronary heart disease (CHD) mortality rates in the community of Goteborg during 1990 to 1995. METHODS: Patients aged <75 years who were hospitalized in the community of Goteborg for AMI during 1990 to 1991 (period 1) and 1995 to 1996 (period 2) were compared in terms of history, treatment for AMI, and outcome. Information on CHD mortality rates in the community of Goteborg was gathered from the National Registry of Deaths. RESULTS: The numbers of patients in the 2 cohorts were 926 and 861, respectively. The incidence rate for AMI per 100,000 inhabitants and year was 200 for period 1 and 183 during period 2. During period 2, there was an increased use of percutaneous transluminal coronary angioplasty, coronary artery bypass grafting, angiotensin-converting enzyme inhibitors, heparin, and intravenous nitroglycerin. On the other hand, there was a decreased use of thrombolytic agents, diuretic agents, digitalis, long-acting nitrates, calcium-channel blockers, and lidocaine. The hospital case-fatality rates were 9.4% during period 1 and 6.0% during period 2 (P =.01). The adjusted risk ratio for period 2 versus period 1 was 0.65, with 95% confidence limits of 0.45 to 0.94. The mortality rate over a period of 3 years was 26.5% during period 1 and 17.8% during period 2 (P <.0001). The adjusted risk ratio for period 2 versus period 1 was 0.67, with 95% confidence limits of 0.54 to 0.82. Among inhabitants aged 30 to 74 years in the community of Goteborg, the CHD mortality rate decreased in 1995 as compared with 1990 (age-adjusted odds ratio 0.79, 95% confidence limits 0.68 to 0.92). CONCLUSIONS: For consecutive patients aged <75 years who were hospitalized for AMI in the community of Goteborg, we found that in the thrombolytic era, major changes in medical and nonmedical treatment still took place associated with a continuing decrease in mortality rates during 3 years of follow-up. A similar reduction of CHD mortality rates was seen in the same age group within the community of Goteborg. PMID- 12094194 TI - A comparison of the Framingham and European Society of Cardiology coronary heart disease risk prediction models in the normative aging study. AB - BACKGROUND: A number of prediction models have been developed in an attempt to accurately identify patients at increased risk of a first coronary heart disease event. We sought to determine the ten-year incidence of coronary heart disease events in a healthy cohort with measurable risk factors, and to compare these results with the predicted number of events by use of both the Framingham and European Society of Cardiology risk prediction models. METHODS: We compared the predicted and observed number of events in 5 risk categories in 1393 subjects aged 30 to 74 years who were enrolled in the Normative Aging Study. RESULTS: The risk prediction models reliably stratify populations with regards to relative risk of coronary heart disease events and there is reasonable agreement between the 2 models (weighted kappa = 0.46, P <.01). The Framingham model underestimated the absolute risk of coronary heart disease events in the low-risk group, and both risk prediction models overestimated the absolute risk of events in the high or very-high-risk groups (Framingham c-statistic = 0.60, European Society of Cardiology c-statistic = 0.58). CONCLUSIONS: Despite simplification, the accuracy of the European model was not significantly different from the Framingham model. But the accuracy of absolute risk prediction, particularly at the extremes of risk, is imperfect. Refinement and validation of these risk prediction models is important because they affect the management of individual patients and the allocation of community resources. PMID- 12094195 TI - Impact of sex and its interaction with age on the management of and outcome for patients with acute myocardial infarction in 4 Japanese hospitals. AB - BACKGROUND: Several studies from the United States and from European countries have detected sex and age differences in clinical characteristics, management, and outcomes of acute myocardial infarction. The aim of this study was to determine how sex and age influence the management of and outcome for patients with acute myocardial infarction in Japan. METHODS: A retrospective cohort study was performed by means of patient chart review at 4 teaching hospitals in Japan. There was a total of 482 patients (136 females [28%], 346 males [72%]) admitted consecutively with a diagnosis of acute myocardial infarction between July, 1995 and June, 1996. RESULTS: Female patients were older and had more comorbid diseases than male patients. Female patients also tended to have more cardiac complications during hospitalization and a greater 30-day mortality (10% vs 4%, P <.05). After adjustment for baseline characteristics and age/sex interaction, it was found that female patients were less likely to undergo thrombolytic therapy, cardiac catheterization, or revascularization, and they had a greater 30-day mortality. These sex differences in cardiac catheterization and revascularization were more pronounced for older patients. On the other hand, the sex differences in 30-day mortality were greater for younger patients. CONCLUSIONS: Our data suggest that cardiac catheterization, revascularization and 30-day mortality may have been related to patient sex and age, but further study is needed. PMID- 12094196 TI - Relationship between vascular dysfunction in peripheral arteries and ischemic episodes during daily life in patients with ischemic heart disease and hypercholesterolemia. AB - BACKGROUND: It is well established that endothelial dysfunction is present in patients with ischemic heart disease and hypercholesterolemia. Some of these patients will have signs of transient myocardial ischemia during Holter monitoring. We sought to describe the correlation between daily life ischemia and signs of endothelial dysfunction as assessed by means of brachial vasoreactivity. METHODS: We included in the study 131 patients with documented ischemic heart disease and a serum cholesterol level of > or =5 mmol/L before the institution of lipid-lowering treatment and dietary intervention. RESULTS: Satisfactory 48-hour Holter recordings and ultrasound scans of the brachial artery were obtained in 119 patients. During 5712 hours of ambulatory monitoring, 181 episodes of transient ST-segment depression with a mean duration of 52 +/- 66 minutes were recorded in 31 patients. The mean percentage dilatation of the brachial artery after occlusion was 4.38% +/- 5.66%; after nitroglycerin administration, it was 13.86% +/- 7.06%. By means of Spearman correlation analysis, the number of ischemic episodes and degree of flow-mediated vasodilatation and nitroglycerin mediated vasodilatation were significantly negatively correlated (r = -0.249, P =.006 and r = -0.302, P =.02, respectively). In a linear regression model, the presence of ischemic episodes was a significant predictor of impaired flow mediated vasodilatation (beta = -3.31, P <.01), even after the adjustment for vessel size and classic cardiovascular risk factors. CONCLUSIONS: These results indicate a significant relationship between ischemic episodes and vascular dysfunction in patients with ischemic heart disease and hypercholesterolemia and may justify an aggressive preventive therapy targeted directly at the endothelium. PMID- 12094197 TI - Testing the test: the reliability of echocardiography in the sequential assessment of valvular regurgitation. AB - BACKGROUND AND OBJECTIVE: Substantial variability in serial echocardiographic qualitative assessment of valvular regurgitation may exist. Reader variability is generally well understood, but acquisition variability (portions of variability caused by equipment, sonographers, physiologic changes) has been less frequently assessed, particularly in combination with reader variability. We attempted to determine the relative contributions of acquisition and reader variability as components of total test-retest variability for aortic (AR) and mitral (MR) regurgitation. METHODS: Outpatient echocardiographic study was done at 2 clinical sites. Twenty-three predominantly obese middle-aged females had 3 echocardiograms, 2 performed 14 +/- 3 days apart and the third performed within 1 to 2 hours of the second. Triplets of echocardiograms were evaluated for change in grade of AR and MR. Medical history, anthropometrics, and blood pressures were obtained. RESULTS: Average intrareader variability (percentage of reads for which there is within-reader disagreement) was 5.6% for AR and 16.7% for MR. The average total test-retest variability (percentage of reads for which there is disagreement between visits) was 29.0% for AR and 24.6% for MR. The acquisition variability for AR was 23.4% +/- 7.7%; for MR, it was 7.9% +/- 10.2%. A significant predictor of change for AR/MR was the initial grade. Change in diastolic blood pressure was positively associated with change in AR and MR. CONCLUSIONS: Intrareader agreement was substantial for AR and MR. Components of total test-retest variability found were reader, biological (change in diastolic blood pressure), and regression to the mean. Recommendations for clinical practice include monitoring blood pressure changes and understanding the confidence limits of the clinical test. Test-retest variability and its components should be considered in echocardiography and other diagnostic testing. PMID- 12094198 TI - Iodine-123 metaiodobenzylguanidine scintigraphic analysis of myocardial sympathetic innervation in patients with AL (primary) amyloidosis. AB - BACKGROUND: Although a high incidence of myocardial adrenergic denervation has been reported in patients with familial amyloid polyneuropathy, assessment of cardiac sympathetic nerve function has not been available in patients with AL (primary) amyloidosis. METHODS: To test the hypothesis that myocardial sympathetic nerve innervation might be impaired and variable according to the presence or absence of clinical autonomic abnormalities and congestive heart failure in AL amyloidosis, we examined 25 patients by use of iodine-123 metaiodobenzylguanidine (MIBG) scintigraphy. RESULTS: Ten of the 16 patients without autonomic symptoms and 5 of the 9 patients with autonomic neuropathy showed congestive heart failure. The heart/mediastinal activity (H/M) ratio (1.53 +/- 0.06 vs 1.29 +/- 0.05 at 3 hours, P <.001) and myocardial washout ratio (41.5% +/- 4.8% vs 30.8% +/- 4.0%, P <.001) of MIBG were significantly increased in patients without autonomic symptoms compared with patients showing autonomic neuropathy. In patient groups with and without autonomic dysfunction, patients demonstrating congestive heart failure exhibited a significantly decreased H/M ratio and increased washout compared with patients with no heart failure, and left ventricular fractional shortening was positively correlated with the H/M ratio and inversely correlated with the washout ratio. There were significant correlations between the low-frequency component of the heart rate variability and the H/M ratio and washout ratio in the entire patient population. CONCLUSIONS: Patients with AL amyloidosis and no autonomic dysfunction showed variable degrees of enhanced cardiac adrenergic neuronal activity with presynaptic sympathetic dysfunction. In contrast, patients with AL amyloidosis and autonomic neuropathy exhibited prominent myocardial adrenergic denervation with normal or impaired sympathetic neural function of the heart. This study demonstrates that myocardial uptake and turnover of MIBG in patients with AL amyloidosis are heterogeneous and dependent on the presence or absence of congestive heart failure and cardiac autonomic dysfunction. PMID- 12094199 TI - Correlates of coronary blood flow before and after percutaneous coronary intervention and their relationship to angiographic and clinical outcomes in the RESTORE trial. Randomized Efficacy Study of Tirofiban for Outcomes and REstenosis. AB - BACKGROUND AND OBJECTIVES: Slower blood flow in the setting of acute myocardial infarction (MI) has been related to adverse outcomes, but the relationship of coronary blood flow after percutaneous transluminal coronary angioplasty (PTCA) in the setting of acute coronary syndromes to adverse outcomes and restenosis has not been well described. We sought to evaluate the correlates of pre- and post PTCA coronary blood flow to shed light on potential modifiable determinants. METHODS: The RESTORE trial (Randomized Efficacy Study of Tirofiban for Outcomes and REstenosis) was a randomized, double-blind, placebo-controlled trial of tirofiban in patients undergoing balloon angioplasty or directional atherectomy within 72 hours of occurrence of either unstable angina pectoris or acute MI. Coronary blood flow was assessed with the corrected TIMI frame count (CTFC), and clinical outcomes were assessed at 30 days. RESULTS: In addition to tighter and longer minimum lumen diameters (MLDs), the multivariate correlates of slower flow before PTCA also included the presence of thrombus, collaterals, left coronary artery lesion location, acute MI, and >8F catheter size. As well as the above variables, type C and D dissection grades were related to slower post-PTCA CTFC. Death, or the composite of death/MI/coronary artery bypass graft at 30 days, was more frequent among patients with slower post-PTCA CTFCs and those with post-PTCA thrombus. In a multivariate model correcting for reference segment diameter and MLD, the post-PTCA CTFC was an independent predictor of late lumen loss and the follow-up MLD at 6 months. As a single index that integrates functional and anatomical aspects of the post-PTCA results, the ratio of CTFC/MLD was associated with death/MI by 30 days. CONCLUSIONS: In addition to MLD, variables such as the presence of thrombus, left coronary artery lesion location, and dissection grade also are associated with slower coronary blood flow after PTCA. In turn, post PTCA CTFCs were an independent predictor of late lumen loss and follow-up MLDs. Furthermore, patients who die or who sustain other adverse cardiac events have slower coronary blood flow and greater thrombus burden after PTCA. PMID- 12094200 TI - High shear stress after successful balloon angioplasty is associated with restenosis and target lesion revascularization. AB - BACKGROUND: Vascular wall shear stress (WSS) has been implied in the pathogenesis of atherosclerosis and vascular remodeling. Our aim was to calculate WSS after balloon angioplasty and evaluate its predictive value for long-term outcome. METHODS: WSS was calculated proximal to, in, and distal to the lesion after angiographically successful balloon dilatation and at follow-up with the Hagen Poiseuille formula (WSS = 4 microQ/piR3) in 202 patients. Volumetric blood flow (Q) and lumen radius (R) were derived from Doppler scanning velocities and videodensitometric cross-sectional areas. RESULTS: Postprocedural proximal and in lesion values were higher in vessels that developed restenosis (n = 72; 1.22 +/- 0.61 N/m2 and 3.61 +/- 2.38 N/m2, respectively) than in those without restenosis (n = 130, 1.05 +/- 0.51 N/m2 and 2.46 +/- 1.39 N/m2, respectively; P <.05). In lesion WSS was revealed to be predictive of restenosis by means of logistic regression analysis (odds ratio [OR] = 1.42, P <.05), whereas WSS of the proximal segment was associated with an increased rate of target lesion revascularization (OR = 2.33, P <.005). In the multivariate regression model, WSS of the proximal segment was the only independent predictor when entered with known predictors such as diameter stenosis and coronary flow reserve (OR = 2.15, P <.05). CONCLUSIONS: WSS in the lesion after balloon angioplasty is predictive of angiographic restenosis. Moreover, the proximal value is an independent predictor of target lesion revascularization. This study opens perspectives for the on-line use of WSS as a combined parameter of anatomy and physiology to assess the necessity of adjunctive therapy after successful balloon dilatation. PMID- 12094201 TI - Effects of hospital volume on long-term outcomes after percutaneous transluminal coronary angioplasty after acute myocardial infarction. AB - BACKGROUND: Volume of procedures has been associated with short-term outcome after percutaneous transluminal coronary angioplasty. However, the effect of hospital procedural volume on long-term outcome after PTCA is unknown. METHODS AND RESULTS: We analyzed the physician claims and discharge data of 6635 patients who underwent PTCA after acute myocardial infarction (AMI) between 1991 and 1995 in the province of Quebec, Canada. For each administrative year, hospitals in which PTCA was performed were classified into 3 groups: low-volume, <200 procedures per year; medium-volume, 200 to 399 procedures per year; and high volume, > or =400 procedures per year. Compared with patients in medium-volume and high-volume hospitals, patients in low-volume hospitals were older, had more recent AMI, and were less likely to have been transferred for PTCA. After adjustment for baseline differences, patients in the low-volume and medium-volume groups were more likely to undergo CABG within 6 months compared with patients in the high-volume group (odds ratio [OR] 2.1, 95% CI 1.3-3.3, and OR 1.5, 95% CI 1.2-1.9, respectively). In contrast, patients in the low-volume and medium-volume groups were less likely than patients in the high-volume group to undergo repeat PTCA within 6 months (OR 0.37, 95% CI 0.24-0.58, and OR 0.8, 95% CI 0.70-0.92, respectively). At 6 months, adjusted rates of repeat revascularization, recurrent AMI, or death did not differ between the 3 groups. CONCLUSION: Overall adverse event rates at 6 months after PTCA do not differ between hospital volume groups. The higher rate of CABG in low-volume hospitals and the higher rate of repeat PTCA in high-volume hospitals may represent different physician preferences for the treatment of failed PTCA rather than higher complication rates. PMID- 12094202 TI - Determination of platelet aggregation inhibition during percutaneous coronary intervention with the platelet function analyzer PFA-100. AB - BACKGROUND: A simple device to rapidly evaluate platelet function may aid in optimizing glycoprotein IIb/IIIa inhibition during percutaneous coronary intervention (PCI). We prospectively studied platelet function in 250 patients receiving abciximab or eptifibatide during PCI. METHODS AND RESULTS: The platelet function analyzer PFA-100 (Dade-Behring, Deerfield, Ill) measures platelet function by determining the time to occlusion of an aperture in a biochemically active membrane as whole blood flows under high shear conditions. Platelet aggregation causes aperture occlusion, and results are reported as a closure time (CT). All patients received either abciximab or eptifibatide, along with aspirin and heparin; patients undergoing stent implantation received aspirin and a thienopyridine postprocedure. The CT was measured at baseline and 10 minutes, 4 hours, 12 hours (abciximab-only), and 24 hours after the bolus. Profound inhibition was exhibited in most patients shortly after the platelet inhibitor bolus and during the course of therapy. We observed recovery of platelet function 12 hours after discontinuation of abciximab, with a high degree of interpatient variability, and ongoing profound platelet inhibition 4 to 6 hours after the discontinuation of eptifibatide. Among patients treated with abciximab, patients who were obese recovered from platelet inhibition sooner than patients who were not obese, whereas patients who were elderly had delayed recovery compared with patients who were not elderly. Failure to achieve maximal platelet inhibition (nonclosure) at 10 minutes indicated a possible association with adverse clinical events at the 6-month follow-up examination (60% vs 20%). CONCLUSIONS: PFA-100 is a rapid simple assay used as a means of assessing inhibition of platelet aggregation during PCI performed with glycoprotein IIb/IIIa inhibition. Failure to achieve nonclosure early after the initiation of abciximab therapy warrants further investigation because there may be an association with adverse cardiac events at 6-month follow-up. PMID- 12094203 TI - Cardiac rhythm and symptomatic arrhythmia in right atrial isomerism. AB - BACKGROUND: The conduction system in right atrial isomerism may be complicated by the presence of paired sinus nodes and twin atrioventricular nodes. With a large cohort of infants and children with right atrial isomerism, we sought to investigate the nature of atrial rhythm, its prevalence, factors predisposing patients to symptomatic cardiac arrhythmia, and the relation of arrhythmia to long-term morbidity and mortality. METHODS: Standard 12-lead baseline electrocardiograms were performed in 110 infants and children with right atrial isomerism at a median age of 1 day, and their clinical records were reviewed. The type, timing, and precipitating factors of symptomatic cardiac arrhythmia that occurred in 15 patients, among a cohort of 85 patients who had or were awaiting surgical interventions, were noted. RESULTS: All patients except 1 had a sinus rhythm with intact atrioventricular conduction. Of these, 87% (95/109) had single P-wave morphology, whereas 13% (14/109) had multiple P-wave morphologies. For patients with a single P-wave morphology, the frontal P-wave axis was between 0 degrees and 90 degrees in 62% (59/95), 90 degrees and 180 degrees in 23% (22/95), and superior in 15% (14/95). There was no relation between either P-wave axis or morphology and cardiac anatomy. Symptomatic cardiac arrhythmia occurred in 15 of 85 patients (18%); 11 of the 15 patients had supraventricular tachycardia, and 1 patient each had atrial tachycardia, atrial flutter, ventricular tachycardia, and congenital complete heart block. The arrhythmias occurred before surgery in 4 patients, early after surgery in 5 patients, and late after surgery in 6 patients. Although 3 of the 7 patients who died suddenly had a history of symptomatic arrhythmia, arrhythmia was the documented cause of mortality in only 1 of the 32 fatalities (3.1%). Freedom from arrhythmia at 1, 5, 10, 15, and 20 years was 93% +/- 3%, 86% +/- 4%, 80% +/- 6%, 73% +/- 9%, and 48% +/- 15% (mean +/- SE), respectively. No risk factors for symptomatic arrhythmia were identified by means of logistic regression. CONCLUSIONS: The atrial pacemaker varies in location within and between patients with right atrial isomerism. Although symptomatic cardiac arrhythmias are not uncommon, they do not seem to relate to the overall high mortality rate and occurrence of sudden death in this patient group. Nonetheless, detailed assessment and aggressive management of cardiac arrhythmias once they occur are warranted because of the precarious single ventricular hemodynamics. PMID- 12094204 TI - A novel mechanism for the beneficial vascular effects of high-density lipoprotein cholesterol: enhanced vasorelaxation and increased endothelial nitric oxide synthase expression. AB - BACKGROUND: Low levels of high-density lipoprotein (HDL) cholesterol increase the risk of coronary artery disease (CAD), and recent clinical studies suggest that interventions in low-HDL patients are beneficial. The purpose of this study was to examine the effect of increased HDL levels on endothelium-dependent vasodilation. METHODS: We studied patients with CAD with a low-density lipoprotein (LDL) level of <100 mg/dL. Patients with an HDL level of < or =36 mg/dL were treated with niacin (n = 11), and patients with an HDL level of >36 mg/dL were followed as controls (n = 10). Baseline and 3-month follow-up studies of flow-mediated dilation (FMD) and blood lipid levels were obtained. RESULTS: HDL levels increased from 30.1 +/- 1.2 to 40.5 +/- 1.2 mg/dL in the niacin treated patients (P <.001) but remained unchanged in the control patients. At baseline, FMD was impaired in both the treated (6.5% +/- 1%) and the control (7.3% +/- 1%) patients compared with 10 healthy subjects (16% +/- 2%, P <.01). After 3 months, FMD improved in the niacin-treated patients (11.8% +/- 1%, P =.001) but remained unchanged in the control patients (6.2% +/- 1%). Exposure of cultured human vascular endothelial cells to HDL in vitro enhanced expression of endothelial nitric oxide synthase (eNOS), as shown by immunoblotting. CONCLUSIONS: In patients with CAD and well-controlled LDL levels, elevation of HDL with niacin improves endothelial function. HDL increases eNOS protein expression in cultured vascular endothelial cells. Taken together, these observations suggest that HDL-mediated increases in eNOS expression may contribute to the observed enhancement in vasorelaxation and thus support a previously unrecognized mechanism for the beneficial cardiovascular effects of HDL. PMID- 12094205 TI - Effect of atrial flutter on exercise tolerance in patients with grown-up congenital heart (GUCH). AB - OBJECTIVES: To assess the effect of atrial flutter (AFL) on exercise tolerance in patients with grown-up congenital heart (GUCH), exercise tests with modified Bruce protocol were performed in 20 patients aged 21 to 62 years with GUCH (11 females, 9 males) during symptomatic AFL and again 24 to 48 hours after DC conversion to sinus rhythm (SR). At the same time, cardiac function was assessed by means of transthoracic Doppler echocardiography. RESULTS: Mean exercise duration was significantly less during AFL (6.4 +/- 4.1 min) versus SR (10.9 +/- 3.7 min) (P <.001). Heart rate was faster at rest and peak exercise while in AFL (106 +/- 21 beats/min vs 77 +/- 14 beats/min, P <.001, and 157 +/- 31 beats/min vs 129 +/- 24 beats/min, P <.01, respectively). Systolic blood pressure was lower at peak exercise with AFL (112 +/- 25 mm Hg vs 137 +/- 24 mm Hg, P <.001), as was mean blood pressure increase (5.3 +/- 24.3 mm Hg vs 22.6 +/- 15.8 mm Hg) compared with SR (P <.01). Four of the 6 patients after Fontan surgery had a decrease of 16 mm Hg in systolic blood pressure at peak exercise when in AFL. The reasons for exercise termination during AFL were mainly breathlessness, chest pain, or presyncope, whereas in SR it was caused by fatigue. Echocardiography during AFL showed shorter isovolumic relaxation time (40 +/- 20 ms) compared with SR (50 +/- 20 ms) (P <.05). Ventricular long-axis excursion was reduced (left 1.0 +/- 0.3 cm vs 1.2 +/- 0.4 cm, septal 0.5 +/- 0.2 cm vs 0.7 +/- 0.3 cm, and right 0.7 +/- 0.2 cm vs 0.9 +/- 0.4 cm respectively, P <.001 for all), as were peak pulmonary and aortic flow velocities (85 +/- 30 cm/s vs 105 +/- 50 cm/s, P <.001, and 137 +/- 118 cm/s vs 143 +/- 114 cm/s, P <.02) compared with sinus rhythm. There was a close correlation between exercise duration and blood pressure increase (r = 0.6), left-sided long-axis excursion and blood pressure increase (r = 0.57), and between aortic flow velocity and right-sided long-axis excursion (r = 0.71). CONCLUSIONS: Atrial flutter causes dramatic reduction in exercise tolerance in patients with GUCH, and the combination of fast heart rate and hypotension may contribute to the development of presyncope, particularly in those with Fontan surgery. Marked improvement in effort tolerance and cardiac dynamics occurs after regaining SR. Thus, improving the quality of life in patients with GUCH requires maintaining SR. PMID- 12094206 TI - Angioplasty for coarctation in different aged patients. AB - BACKGROUND: Differences in the indication and outcome of balloon angioplasty for coarctation in children and adults have not been elucidated sufficiently. The results of balloon angioplasty for coarctation are compared between pediatric and adult age groups. METHODS: Balloon angioplasty for coarctation of the aorta was performed in 85 patients who were classified according to age and native coarctation/recoarctation. Groups A (patients aged <16 years, n = 32) and B (patients aged > or =16 years, n = 17) included patients with native coarctations. Groups rCoA A (patients aged <16 years, n = 33) and rCoA B (patients aged > or =16 years, n = 3) included patients with recoarctations. Follow-up included 2-dimensional Doppler scanning echocardiography and additional angiography or magnetic resonance imaging. Gradient reductions in groups were compared by use of the independent-samples t test. Kaplan-Meier and log-rank analyses were performed as a means of comparing long-term outcome. RESULTS: No mortality occurred. Immediate success was equal in groups A, B, and rCoA A (94%). Dilatation was unsuccessful in 2 patients in group rCoA B. Pressure gradients decreased 23 mm Hg in group A, 31 mm Hg in group B, 18 mm Hg in group rCoA A, and 11 mm Hg in group rCoA B. Pressure gradient drops, compared between groups A and B, showed a significant difference (P <.001). The length of hospital stay ranged from 12 to 48 hours. The period of follow-up ranged from 6 months to 12 years (mean, 4.9 years). Kaplan-Meier curves of groups A and B are not different, as determined by means of log-rank analysis. No aneurysm formation was encountered. CONCLUSIONS: The results of balloon angioplasty for native coarctation in both selected children and adults are excellent. In recoarctation, we recommend balloon angioplasty in the pediatric patients. PMID- 12094207 TI - Diversification of synaptic strength: presynaptic elements. AB - Synapses are not static; their performance is modified adaptively in response to activity. Presynaptic mechanisms that affect the probability of transmitter release or the amount of transmitter that is released are important in synaptic diversification. Here, we address the diversity of presynaptic performance and its underlying mechanisms: how much of the variation can be accounted for by variation in synaptic morphology and how much by molecular differences? Significant progress has been made in defining presynaptic structural contributions to synaptic strength; by contrast, we know little about how presynaptic proteins produce normally observed functional differentiation, despite abundant information on presynaptic proteins and on the effects of their individual manipulation. Closing the gap between molecular and physiological synaptic diversification still represents a considerable challenge. PMID- 12094208 TI - Proneural genes and the specification of neural cell types. AB - Certain morphological, physiological and molecular characteristics are shared by all neurons. However, despite these similarities, neurons constitute the most diverse cell population of any organism. Recently, considerable attention has been focused on identifying the molecular mechanisms that underlie this cellular diversity. Parallel studies in Drosophila and vertebrates have revealed that proneural genes are key regulators of neurogenesis, coordinating the acquisition of a generic neuronal fate and of specific subtype identities that are appropriate for the location and time of neuronal generation. These studies reveal that, in spite of differences between invertebrate and vertebrate neural lineages, Drosophila and vertebrate proneural genes have remarkably similar roles. PMID- 12094209 TI - Specification of catecholaminergic and serotonergic neurons. AB - The specification of neurotransmitter phenotype is an important aspect of neuronal fate determination. Substantial progress has been made in uncovering key extracellular signals and transcriptional regulators that control the mode of neurotransmission in several model systems, among which catecholaminergic and serotonergic neurons feature prominently. Here, we review our current knowledge of the regulatory circuits that direct neurotransmitter choice, and discuss the development of well-studied types of catecholaminergic and serotonergic neurons. One emerging concept is that different types of neuron use a similar core programme to control shared modes of neurotransmission, but recruit different factors that are specific for each neuronal type. Another is that most factors that specify neurotransmitter identity also control other features of the neuronal phenotype. PMID- 12094210 TI - Memory use in insect visual navigation. AB - The navigational strategies that are used by foraging ants and bees to reach a goal are similar to those of birds and mammals. Species from all these groups use path integration and memories of visual landmarks to navigate through familiar terrain. Insects have far fewer neural resources than vertebrates, so data from insects might be useful in revealing the essential components of efficient navigation. Recent work on ants and bees has uncovered a major role for associative links between long-term memories. We emphasize the roles of these associations in the reliable recognition of visual landmarks and the reliable performance of learnt routes. It is unknown whether such associations also provide insects with a map-like representation of familiar terrain. We suggest, however, that landmarks act primarily as signposts that tell insects what particular action they need to perform, rather than telling them where they are. PMID- 12094211 TI - A common reference frame for movement plans in the posterior parietal cortex. AB - Orchestrating a movement towards a sensory target requires many computational processes, including a transformation between reference frames. This transformation is important because the reference frames in which sensory stimuli are encoded often differ from those of motor effectors. The posterior parietal cortex has an important role in these transformations. Recent work indicates that a significant proportion of parietal neurons in two cortical areas transforms the sensory signals that are used to guide movements into a common reference frame. This common reference frame is an eye-centred representation that is modulated by eye-, head-, body- or limb-position signals. A common reference frame might facilitate communication between different areas that are involved in coordinating the movements of different effectors. It might also be an efficient way to represent the locations of different sensory targets in the world. PMID- 12094212 TI - The amygdala and reward. AB - The amygdala -- an almond-shaped group of nuclei at the heart of the telencephalon -- has been associated with a range of cognitive functions, including emotion, learning, memory, attention and perception. Most current views of amygdala function emphasize its role in negative emotions, such as fear, and in linking negative emotions with other aspects of cognition, such as learning and memory. However, recent evidence supports a role for the amygdala in processing positive emotions as well as negative ones, including learning about the beneficial biological value of stimuli. Indeed, the amygdala's role in stimulus-reward learning might be just as important as its role in processing negative affect and fear conditioning. PMID- 12094213 TI - Animal models of neurological deficits: how relevant is the rat? AB - Animal models of neurological deficits are essential for the assessment of new therapeutic options. It has been suggested that rats are not as appropriate as primates for the symptomatic modelling of disease, but a large body of data argues against this view. Comparative analyses of movements in rats and primates show homology of many motor patterns across species. Advances have been made in identifying rat equivalents of akinesia, tremor, postural deficits and dyskinesia, which are relevant to Parkinson's disease. Rat models of hemiplegia, neglect and tactile extinction are useful in assessing the outcome of ischaemic or traumatic brain injury, and in monitoring the effects of therapeutic interventions. Studies in rodents that emphasize careful behavioural analysis should continue to be developed as effective and inexpensive models that complement studies in primates. PMID- 12094214 TI - Mechanisms and functions of Eph and ephrin signalling. PMID- 12094215 TI - Remodelling the walls of the nucleus. PMID- 12094216 TI - Synaptotagmin: a Ca(2+) sensor that triggers exocytosis? PMID- 12094217 TI - Protease degradomics: a new challenge for proteomics. PMID- 12094218 TI - A greener world: the revolution in plant bioimaging. PMID- 12094219 TI - Opinion: Building epithelial architecture: insights from three-dimensional culture models. AB - How do individual cells organize into multicellular tissues? Here, we propose that the morphogenetic behaviour of epithelial cells is guided by two distinct elements: an intrinsic differentiation programme that drives formation of a lumen enclosing monolayer, and a growth factor-induced, transient de-differentiation that allows this monolayer to be remodelled. PMID- 12094220 TI - The strange case of the signal recognition particle. PMID- 12094221 TI - BAFF: a fundamental survival factor for B cells. AB - B-cell-activating factor of the tumour-necrosis-factor family (BAFF) enhances B cell survival--a function that is indispensable for B-cell maturation--and has a role in enhancing immune responses. Moreover, the overexpression of BAFF results in severe autoimmune disorders in mice, and elevated serum levels of BAFF occur in some patients who have autoimmune diseases. The elucidation of the role of BAFF has set the stage for a new approach to the treatment of autoimmune disease. PMID- 12094222 TI - VAV proteins as signal integrators for multi-subunit immune-recognition receptors. AB - In recent years, substantial progress has been made towards the identification of intracellular signalling molecules that couple multi-subunit immune-recognition receptors (MIRRs) to their various effector functions. Among these, the VAV proteins have been observed to have a crucial role in regulating some of the earliest events in receptor signalling. VAV proteins function, in part, as guanine-nucleotide exchange factors (GEFs) for the RHO/RAC family of GTPases. This review focuses on the role of VAV proteins in the regulation of lymphocyte development and function, and emphasizes the regulatory roles that these proteins have through both GEF-dependent and -independent mechanisms. PMID- 12094223 TI - Selection and fine-tuning of the autoimmune T-cell repertoire. AB - The immune system must avoid aggressive T-cell responses against self-antigens. But, paradoxically, exposure to self-peptides seems to have an important role in positive selection in the thymus and the maintenance of a broad T-cell repertoire in the periphery. Recent experiments have highlighted situations that allow high avidity self-reactive T cells to avoid negative selection in the thymus. Accumulating evidence indicates that other, non-deleting mechanisms control the avidity with which T cells recognize self-antigens--a phenomenon that is known as 'tuning'. This might maximize the peripheral T-cell repertoire by allowing the survival of T cells that can respond to self, but only at concentrations that are not normally reached in vivo. PMID- 12094224 TI - The immunobiology of schistosomiasis. AB - Schistosomes are parasitic worms that are a prime example of a complex multicellular pathogen that flourishes in the human host despite the development of a pronounced immune response. Understanding how the immune system deals with such pathogens is a daunting challenge. The past decade has seen the use of a wide range of new approaches to determine the nature and function of the immune response to schistosomes. Here, we attempt to summarize advances in our understanding of the immunology of schistosomiasis, with the bulk of the review reflecting the experimental focus on Schistosoma mansoni infection in mice. PMID- 12094225 TI - T-cell-receptor gene therapy. AB - T cells are tightly controlled cellular machines that monitor changes in epitope presentation. Although T-cell function is regulated by means of numerous interactions with other cell types and soluble factors, the T-cell receptor (TCR) is the only structure on the T-cell surface that defines its antigen-recognition potential. Consequently, the transfer of T-cell receptors into recipient cells can be used as a strategy for the passive transfer of T-cell immunity. In this review, I discuss the pros and cons of TCR gene transfer as a strategy to induce defined virus- and tumour-specific T-cell immunity. PMID- 12094226 TI - Smallpox: anything to declare? AB - Smallpox was eradicated in 1977, but it remains a concern owing to the potential use of the causative agent variola virus in bioterrorism. This article provides an overview of the World Health Organization's spectacular success in achieving the eradication of smallpox. It discusses how variola virus could potentially re emerge and how prepared we are to counter such a re-emergence. Finally, the potential threat from other orthopoxviruses that exist naturally or that have been genetically engineered is considered. In the words of Rep. Christopher Shay, 'Better to be scared by the improbable possibility than to be unprepared for the catastrophic reality'. PMID- 12094227 TI - Apoptosis as a therapeutic tool in rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a chronic inflammatory synovitis that is dominated by the presence of macrophages, lymphocytes and synovial fibroblasts, which leads to the destruction of bone and cartilage. The effectiveness of therapies that are directed against tumour-necrosis factor and interleukin-1 has identified macrophages as a crucial target for therapeutic intervention. However, not all patients respond to these therapies, and the benefits of this form of treatment are short lived. Recent work indicates that the insufficient apoptosis of inflammatory cells in the RA joint might contribute to pathogenesis. In this article, I characterize the mechanisms that prevent the apoptosis of chronic inflammatory cells in the RA joint, to identify potential new targets for the treatment of RA. PMID- 12094228 TI - Regulatory phases of early liver development: paradigms of organogenesis. AB - Genetic analysis, embryonic tissue explantation and in vivo chromatin studies have together identified the distinct regulatory steps that are necessary for the development of endoderm into a bud of liver tissue and, subsequently, into an organ. In this review, I discuss the acquisition of competence to express liver specific genes by the endoderm, the control of early hepatic growth, the coordination of hepatic and vascular development and the cell differentiation that is necessary to generate a functioning liver. The regulatory mechanisms that underlie these phases are common to the development of many organ systems and might be recapitulated or disrupted during stem-cell differentiation and adult tissue pathogenesis. PMID- 12094229 TI - Organogenesis: molecular mechanisms of tubulogenesis. AB - As organisms have evolved in size and complexity, tubular systems have developed to enable the efficient transport of substances into and out of tissues. These tubular systems are generated using strategies that are based on common elements of cell behaviour, including cell polarization, tube migration to target sites, cell-fate diversification and localization of specialized cells to different regions of the tube system. Using examples from both invertebrate and vertebrate systems, this review highlights progress in understanding these basic principles and briefly discusses the possible evolution of strategies to regulate the morphogenesis of tubular systems. PMID- 12094230 TI - Pancreatic organogenesis--developmental mechanisms and implications for therapy. AB - The pancreas is a mixed exocrine and endocrine gland that controls many homeostatic functions. The exocrine pancreas produces and secretes digestive enzymes, whereas the endocrine compartment consists of four distinct hormone producing cell types. Studies that further our knowledge of the basic mechanisms that underlie the formation of the pancreas will be crucial for understanding the development and homeostasis of this organ and of the mechanisms that cause diabetes. This information is also pivotal for any attempt to generate functional insulin-producing beta-cells that are suitable for transplantation. PMID- 12094231 TI - Coordinating early kidney development: lessons from gene targeting. AB - The kidney is widely used to study the mechanisms of organogenesis. Its development involves fundamental processes, such as epithelial branching, induced morphogenesis and cytodifferentiation, which are common to the development of many other organs. Gene-targeting experiments have greatly improved our understanding of kidney development, and have revealed many important genes that regulate early kidney organogenesis, some of which have a role in inherited human kidney disorders. Although our understanding of how the kidney is assembled is still limited, these studies are beginning to provide insights into the genetic and cellular interactions that regulate early organogenesis. PMID- 12094232 TI - Patterning the vertebrate heart. AB - The mammalian heart is crafted from a few progenitor cells that are subject to rapidly changing sets of instructions from their environment and from within. These instructions cause them to migrate, expand and diversify in lineage, and acquire form and function. Molecular information from various model systems, combined with increasingly detailed morphogenetic data, has provided insights into some of these key events. Many congenital heart abnormalities might arise from defects in the early stages of heart development, therefore it is important to understand the molecular pathways that underlie the lineage specification and patterning processes that shape this organ. PMID- 12094233 TI - Training for genetic counsellors. AB - Genetic counsellors are uniquely trained to provide support, explanations and guidance to individuals or families who have been diagnosed with a genetic disorder. As our knowledge of the genetic basis of disease increases, so does our ability to diagnose it and so does the demand for appropriately trained genetic counsellors. Despite this growing demand, only a handful of countries provides formal courses in genetic counselling, whereas other countries leave genetic counselling in the hands of medical practitioners or medical geneticists. PMID- 12094234 TI - Genetics education for primary-care providers. AB - In an era of growing knowledge about genetics and health, primary-care physicians will have increasing responsibility for evaluating genetic risk and using genetic tests. Although most have little knowledge of genetics, their expertise in the prudent use of technology is relevant to the task. Successful educational programmes will need to forge partnerships between primary care and genetics. PMID- 12094235 TI - The phosphatidylinositol 3-Kinase AKT pathway in human cancer. PMID- 12094236 TI - Core-binding factors in haematopoiesis and leukaemia. PMID- 12094237 TI - Identification of tumour-associated T-cell epitopes for vaccine development. PMID- 12094238 TI - Roles of heparan-sulphate glycosaminoglycans in cancer. PMID- 12094239 TI - LKB1--a master tumour suppressor of the small intestine and beyond. PMID- 12094240 TI - Chemoprevention: an essential approach to controlling cancer. AB - Mortality that results from the common forms of cancer is still unacceptably high. Despite immense advances in the understanding of the mechanisms of carcinogenesis, in bringing potent new drugs to the clinic and in treating several relatively rare forms of cancer, overall mortality statistics are unlikely to change in a fundamental way until there has been a re-orientation of emphasis in cancer research that will direct greater resources towards prevention of new disease, rather than treatment of end-stage disease. PMID- 12094241 TI - Cancer after nuclear fallout: lessons from the Chernobyl accident. PMID- 12094244 TI - Tyrosine kinase fusion genes in chronic myeloproliferative diseases. AB - With the exception of chronic myeloid leukemia (CML), chronic myeloproliferative disorders (CMPDs) are a heterogeneous spectrum of conditions for which the molecular pathogenesis is not well understood. Most cases have a normal or aneuploid karyotype, but a minority present with a reciprocal translocation that disrupts specific tyrosine kinase genes, most commonly PDGFRB or FGFR1. These translocations result in the production of constitutively active tyrosine kinase fusion proteins that deregulate hemopoiesis in a manner analogous to BCR-ABL. With the advent of targeted signal transduction therapy, an accurate clinical and molecular diagnosis of CMPDs has become increasingly important. Currently, patients with PDGFRB or ABL fusion genes are candidates for treatment with Imatinib (STI571), but it is likely that alternative strategies will be necessary for the treatment of most other patients. PMID- 12094245 TI - Insights from pre-clinical studies for new combination treatment regimens with the Bcr-Abl kinase inhibitor imatinib mesylate (Gleevec/Glivec) in chronic myelogenous leukemia: a translational perspective. AB - Clinical phase I/II studies with the Abl kinase inhibitor imatinib mesylate (Gleevec/Glivec, formerly STI571) for the treatment for chronic myelogenous leukemia (CML) demonstrated the safety and the remarkable efficacy of this molecularly targeted agent. However, a significant proportion of patients treated in the chronic phase of the disease after having failed interferon alpha (IFN) remain predominantly Philadelphia chromosome positive (Ph(+)), suggesting a risk of later relapses. Furthermore, results in blast crisis patients revealed a high frequency of relapses or resistance to imatinib. To circumvent resistance, improve response rates, or prolong survival, pre-clinical evaluations of combinations of imatinib with other agents have been pursued. Some of these have already been translated into clinical studies. Here, we first summarize evidence from pre-clinical studies on new combination regimens with imatinib in the treatment of CML. Second, we analyze preliminary clinical data of ongoing combination studies. Finally, we provide a summary of approaches that use novel antileukemic agents with molecularly characterized modes of action. PMID- 12094246 TI - Safety and efficacy of STI-571 (imatinib mesylate) in patients with bcr/abl positive chronic myelogenous leukemia (CML) after autologous peripheral blood stem cell transplantation (PBSCT). AB - We examined safety and efficacy of STI-571 in 24 bcr/abl-positive patients with CML post PBSCT. At start of STI-571 therapy, nine patients presented in blast crisis (BC) or in accelerated phase (AP), and 15 in chronic phase (CP). Patients were evaluated for hematologic, cytogenetic and molecular response, survival and toxicity. In general, STI-571 was well tolerated in this heavily pretreated group of patients with a non-hematologic and hematologic toxicity profile similar to that observed in a previous phase I trial at comparable doses. Five of nine patients with CML in transformation (AP, BC) were evaluable for hematologic response. Two of five patients had transient reductions in WBC and blasts, and three patients achieved a sustained hematologic response (>4 weeks). Cytogenetic analysis in these patients revealed numerical and/or structural responses. In CML chronic phase, STI-571 induced complete hematologic responses in all patients and major cytogenetic responses in 61% of patients with a complete cytogenetic response rate of 46%. This report indicates that STI-571 is a safe and effective drug in heavily pretreated patients. No apparent additional side-effects were noted in this patient cohort. The high rate of complete hematologic and complete cytogenetic responses in CP patients is remarkable, as intensive treatment approaches plus IFN-alpha failed to be efficient in achieving long-term stabilization of CML in this patient cohort. PMID- 12094247 TI - Chromosomal imbalances in familial chronic lymphocytic leukaemia: a comparative genomic hybridisation analysis. AB - A subset of B cell chronic lymphocytic leukaemia (CLL) is familial. Lack of large families makes it attractive to exploit methods in addition to genetic linkage analysis for the identification of a susceptibility locus. One strategy that can localise regions of the genome that may harbour tumour suppressor genes is to identify regions of chromosomal imbalance using comparative genomic hybridisation (CGH) analysis. We examined 24 familial CLL cases by CGH analysis. Losses that are documented as arising frequently in sporadic CLL were observed at a comparable frequency in familial CLL. However, gains and losses in two regions of the X chromosome - Xp11.2-p21 and Xq21-qter - appear more common in familial CLL than in sporadic CLL. This suggests these regions may harbour a susceptibility locus for CLL. There is also some evidence that chromosome regions 2p12-p14 and 4q11-q21 may harbour predisposition genes. PMID- 12094248 TI - Antigen expression patterns reflecting genotype of acute leukemias. AB - Multi-parameter flow cytometry, molecular genetics, and cytogenetic studies have all contributed to new classification of leukemia. In this review we discuss immunophenotypic characteristics of major genotypic leukemia categories. We describe immunophenotype of: B-lineage ALL with MLL rearrangements, TEL/AML1, BCR/ABL, E2A/PBX1 translocations, hyperdiploidy, and myc fusion genes; T-ALL with SCL gene aberrations and t(5;14) translocation; and AML with AML1/ETO, PML/RARalpha, OTT/MAL and CBFbeta/MYH11 translocations, trisomies 8 or 11 and aberrations of chromosomes 7 and 5. Whereas some genotypes associate with certain immunophenotypic features, others can present with variable immunophenotype. Single molecules (as NG2, CBFbeta/SMMHC and PML/RARalpha proteins) associated with or derived from specific translocations have been described. More often, complex immunophenotype patterns have been related to the genotype categories. Most known associations between immunophenotype and genotype have been defined empirically. Therefore, these associations should be validated in independent patient cohorts before they can be widely used for prescreening of leukemia. Progress in our knowledge on leukemia will show how the molecular-genetic changes modulate the immunophenotype as well as how the expressed protein molecules further modulate cell behavior. PMID- 12094249 TI - Induction therapy by frequent administration of doxorubicin with four other drugs, followed by intensive consolidation and maintenance therapy for adult acute lymphoblastic leukemia: the JALSG-ALL93 study. AB - In order to improve the disappointing prognosis of adult patients with acute lymphoblastic leukemia (ALL), we applied similar induction therapy as that used for acute myeloid leukemia (AML), ie frequent administration of doxorubicin (DOX). DOX 30 mg/m(2) was administered from days 1 to 3 and from days 8 to 10 together with vincristine, prednisolone, cyclophosphamide and L-asparaginase, followed by three courses of consolidation and four courses of intensification. From December 1993 to February 1997, 285 untreated adult patients with de novo ALL were entered. Of 263 evaluable patients (age 15 to 59; median 31), 205 (78%) obtained complete remission (CR). At a median follow-up period of 63 months, the predicted 6-year overall survival (OS) rate of all patients was 33%, and disease free survival (DFS) rate of CR patients was 30%, respectively. By multivariate analysis, favorable prognostic factors for the achievement of CR were age <40 and WBC <50 000/microl; for longer OS were age <30 and WBC <30 000/microl; and for longer DFS of CR patients were FAB L1 and ALT <50 IU/l. Among 229 patients who had adequate cytogenetic data, 51 (22%) had Philadelphia (Ph) chromosome. Ph negative chromosome was a common favorable prognostic factor for CR, longer OS and DFS. DFS was not different between early sequential intensification (n = 48) and intermittent intensification (n = 43) during the maintenance phase. Among CR patients under 40 years old, the 6-year survival was not different between the allocated related allo-BMT group (34 patients) and the allocated chemotherapy group (108 patients). However, among patients with Ph-positive ALL, the survival of patients who actually received allo-BMT was superior to that of patients who received chemotherapy (P = 0.046). PMID- 12094250 TI - Deletion analysis of chromosome 13q14.3 and characterisation of an alternative splice form of LEU1 in B cell chronic lymphocytic leukemia. AB - Heterozygous and homozygous deletions of chromosome 13q14.3 are found in 50% of patients with B cell CLL, suggesting the presence of one or more tumour suppressor genes within the deleted region. To identify candidate genes from the region, we constructed a map of 13q14.3 using a combination of genomic and cDNA library screening. The incidence of deletions in CLL patients was 51.5% encompassing a 265 kb region of minimal deletion (RMD) telomeric to markers D13S319. Two CpG islands were identified within the RMD, the telomeric of which is fully methylated whilst the more centromeric is unmethylated. A novel transcript was identified within the RMD that represents an alternative splice version of Leu1. The nine exons of this transcript span a genomic of 436 kb with exon 1 of Leu1 being the common first exon. The remaining exons were shown to be more frequently deleted than Leu1 itself. All splice forms of this transcript were detectable by RT-PCR but Leu1 detected the most abundant message on Northern blotting. Sequence analysis failed to reveal inactivating mutations in patients with heterozygous deletion of 13q14.3, although a polymorphic T to A variant was identified within exon 1 of Leu1 in leukemic and normal controls. As no mutations have been detected for Leu1 or any other transcript so far described, we cannot exclude the existence of control elements within the RMD that may regulate expression of genes lying in this region. PMID- 12094251 TI - Signals mediated by FcgammaRIIA suppress the growth of B-lineage acute lymphoblastic leukemia cells. AB - We examined Fc receptor expression and function in normal and leukemic human immature B cells. Fc receptor expression increased with normal B cell maturation: CD32(+) cells composed 8.1% +/- 1.2% (mean +/- s.d.) of the least mature (CD34(+)CD10(+)), 19.2% +/- 5.7% of intermediate (CD34(-)CD10(+)), and 82.4% +/- 5.0% of mature (CD34(-)CD10(-)) bone marrow CD19(+) B cells. Forty-five of 57 primary B-lineage acute lymphoblastic leukemia samples and all six cell lines studied expressed Fc receptors. By RT-PCR and antibody staining, FcgammaRIIA was the Fc receptor predominantly expressed in these cells. FcgammaRIIA ligation in RS4;11 and 380 cells induced tyrosine phosphorylation of CD32, CD19, CBL, SYK, P13-K p85 and SHIP, as well as RasGAP association with tyrosine-phosphorylated p62(dok). These signalling events resulted in a marked suppression of leukemia cell growth. After a 7-day exposure to anti-CD32, the recovery of ALL cells cocultured with stroma was reduced to 5.5% +/- 2.8% of control values in 380 cells (n = 14), 19.4% +/- 6.1% (n = 8) in RS4;11, and 4.0% +/- 1.3% (n = 6) in KOPN55bi. CD32 ligation also reduced cell recovery in five of seven CD32(+) primary leukemia samples. Thus, FcgammaRIIA mediates signals that suppress the growth of lymphoid leukemia cells. PMID- 12094252 TI - Non-DNA-binding Ikaros isoform gene expressed in adult B-precursor acute lymphoblastic leukemia. AB - Ikaros, a zinc finger transcription factor, is essential for lymphoid development. Mutant mice expressing dominant-negative Ikaros gene (Ikaros) isoforms develop an aggressive form of lymphoid malignancies. We examined the expression of Ikaros isoforms in 11 leukemic cell lines and adult acute lymphoblastic leukemia cells from 36 patients with B-precursor acute lymphoblastic leukemia (pre-B ALL) and nine with T-precursor acute lymphoblastic leukemia (pre-T ALL), using reverse transcriptase-polymerase chain reaction (RT PCR) analysis. In one pre-B ALL cell line, INC cells, and primary leukemic cells from 16 patients with pre-B ALL, we found the predominant expression of a non-DNA binding Ikaros isoform, Ik-6. However, Ik-6 was not detected in pre-T ALL cells. All of pre-B ALL cells expressing Ik-6 were CD10(+), whereas CD10(-) pre-B ALL cells did not express Ik-6. The expression of Ik-6 was not related to karyotype abnormalities such as t(9;22) and t(4;11). Proteins from the cells that expressed Ik-6 alone failed to bind to the Ikaros protein-specific binding sequence in DNA. Ikaros proteins lacking the DNA binding sequences were detected in the cytoplasm but not in the nucleus of the cells. When INC and primary pre-B ALL cells that express Ik-6 alone were irradiated and cultured in the absence of serum, these cells produced functional Ikaros isoforms, Ik-1 and Ik-2. Purified CD19(+) CD10( ) and CD19(+) CD10(+) cells from normal human bone marrow did not express Ik-6. The predominant expression of Ik-6, which is the result of post-transcription dysregulation, is characteristic of adult pre-B ALL, especially CD10(+) pre-B ALL. PMID- 12094253 TI - Expression pattern of HOXB6 homeobox gene in myelomonocytic differentiation and acute myeloid leukemia. AB - Homeobox genes encode transcription factors known to be important morphogenic regulators during embryonic development. An increasing body of work implies a role for homeobox genes in both hematopoiesis and leukemogenesis. In the present study we have analyzed the role of the homeobox gene, HOXB6, in the program of differentiation of the myeloid cell lines, NB4 and HL60. HOXB6 expression is transiently induced during normal granulocytopoiesis and monocytopoiesis, with an initial induction during the early phases of differentiation, followed by a blockade of expression at early maturation. The enforced expression of HOXB6 in promyelocytic NB4 cells or in myeloblastic HL60 cells elicited inhibition of the granulocytic or monocytic maturation, respectively. Furthermore, HOXB6 was frequently expressed (18 out of 49 cases) in AMLs lacking major translocations while it was expressed at very low frequency (two out of 47 cases) in AMLs characterized by PML/RAR-alpha, AML-1/ETO, CBFbeta/MYH11 fusion and rearrangements of the MLL gene at 11q23. According to these observations, we suggest that a regulated pattern of HOXB6 expression is required for normal granulopoiesis and monocytopoiesis. Abnormalities of the HOXB6 expression may contribute to the development of the leukemic phenotype. PMID- 12094255 TI - Early lymphocyte recovery is a predictive factor for prolonged survival after autologous hematopoietic stem cell transplantation for acute myelogenous leukemia. AB - Absolute lymphocyte count (ALC) recovery correlates with survival after autologous hematopoietic stem cell transplantation (AHSCT) for patients with multiple myeloma, non-Hodgkin's lymphoma, and metastatic breast cancer. The role of ALC recovery in relationship to clinical outcome after AHSCT in patients with acute myelogenous leukemia is unknown. We analyzed 45 patients who underwent AHSCT at Mayo Clinic, Rochester, Minnesota between 1990 and 2000. The ALC threshold was selected at 500 cells/microl on day 15 post-AHSCT based on our previous studies. Thirty-two females and 13 males were included in the study with a median age of 45 years (range 12-75). The median follow-up was 14 months with a maximum of 129 months. The median overall and leukemia-free survival were significantly better for the 23 patients with ALC at day 15 > or =500 cells/microl compared with 22 patients with ALC <500 cells/microl (not yet reached vs 10 months, P < 0.0009; 105 vs 9 months, P < 0.0008, respectively). In conclusion, ALC > or =500 cells/microl on day 15 post-AHSCT is associated with better survival in acute myelogenous leukemia and requires further studies. PMID- 12094254 TI - Overexpression of vascular endothelial growth factor (VEGF) and its cellular receptor KDR (VEGFR-2) in the bone marrow of patients with acute myeloid leukemia. AB - Vascular endothelial growth factor (VEGF) and its cellular receptor VEGFR-2 have been implicated as the main endothelial pathway required for tumor neovascularization. However, the importance of the VEGF/VEGFR-2 system for angiogenesis in hematologic malignancies such as AML remains to be elucidated. In 32 patients with newly diagnosed untreated AML, we observed by immunohistochemical analysis of bone marrow biopsies significantly higher levels of VEGF and VEGFR-2 expression than in 10 control patients (P <0.001). In contrast, VEGFR-1 staining levels in AML patients were in the same range as in the controls. Expression of VEGF and VEGFR-2 was significantly higher in patients with a high degree of microvessel density compared to those with a low degree (VEGF: P =0.024; VEGFR-2: P =0.040) and correlated well with bone marrow microvessel density (r(s)=0.566 and 0.609, respectively; P <0.001). Furthermore, in patients who achieved a complete remission following induction chemotherapy VEGFR-2 staining levels decreased into the normal range. In conclusion, our results provide evidence for increased expression of VEGF/VEGFR-2 of leukemic blasts and correlation with angiogenesis in the bone marrow of AML patients. Thus, VEGF/VEGFR-2 might constitute promising targets for antiangiogenic and antileukemic treatment strategies in AML. PMID- 12094256 TI - Optimized lentiviral transduction of erythroid precursors from healthy adults and patients with myelodysplastic syndromes. AB - Lentivectors, derived from human immunodeficiency virus-1 (HIV-1), represent a novel investigational and therapeutic tool for targeting hematopoietic progenitor cells. We describe a new protocol whereby we achieved a highly efficient lentiviral transduction of erythroid precursor cells originating from the bone marrow of healthy adults and patients with myelodysplastic syndromes (MDS). CD34(+) stem cells from healthy subjects were cultured with erythropoietin, IL-3 and stem cell factor, and thereby expanded approximately 300-fold. When these cultures were transduced with a lentiviral vector expressing GFP as a reporter gene, 70% glycophorin(+) cells were GFP(+). Although proliferation and levels of transduction were reduced in cultures of CD34(+) stem cells from patients with myelodysplastic syndromes, 50% of glycophorin(+) cells became GFP(+), amongst which 30% were sideroblastic erythroid precursors. This study demonstrates that lentiviral vectors are capable of efficiently transducing MDS precursors and offers new perspectives to investigate the influence of specific genes on normal erythroid differentiation. This may eventually help to correct defects in patients suffering from myelodysplastic syndromes. PMID- 12094257 TI - mRNA-electroporated mature dendritic cells retain transgene expression, phenotypical properties and stimulatory capacity after cryopreservation. AB - Genetically modified dendritic cells (DC) are increasingly used in vitro to activate cytotoxic T lymphocyte (CTL) immune responses. Because T cell activation protocols consist of multiple restimulation cycles of peripheral blood lymphocytes with antigen-loaded mature DC, continuous generation of DC is needed throughout the experiment. Therefore, cryopreservation of DC loaded with antigen is a valuable alternative for weekly generation and modification of DC. Recently, we described an antigen loading method for DC based on electroporation of defined tumor antigen mRNA. In this study, we demonstrate that mRNA-electroporated DC can efficiently be prepared for cryopreservation. Using an optimized maturation and freezing protocol after mRNA electroporation, we obtained high transgene expressing viable mature DC. In addition, we showed that these modified cryopreserved DC retain stimulatory capacity in an influenza model system. Therefore, cryopreservation of mature mRNA-electroporated DC is a useful method for continuous availability of antigen-loaded DC throughout T cell activation experiments. PMID- 12094258 TI - Synergistic induction of mitochondrial damage and apoptosis in human leukemia cells by flavopiridol and the histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA). AB - Interactions between the histone deacetylase inhibitor SAHA (suberoylanilide hydroxamic acid) and the cyclin-dependent kinase (CDK) inhibitor flavopiridol (FP) were examined in human leukemia cells. Simultaneous exposure (24 h) of myelomonocytic leukemia cells (U937) to SAHA (1 microM) and FP (100 nM), which were minimally toxic alone (1.5 +/- 0.5% and 16.3 +/- 0.5% apoptosis respectively), produced a dramatic increase in cell death (ie 63.2 +/- 1.9% apoptotic), reflected by morphology, procaspase-3 and -8 cleavage, Bid activation, diminished DeltaPsi(m), and enhanced cytochrome c release. FP blocked SAHA-mediated up-regulation of p21(CIP1) and CD11b expression, while inducing caspase-dependent Bcl-2 and pRb cleavage. Similar interactions were observed in HL-60 and Jurkat leukemic cells. Enhanced apoptosis in SAHA/FP-treated cells was accompanied by a marked reduction in clonogenic surivival. Ectopic expression of either dominant-negative caspase-8 (C8-DN) or CrmA partially attenuated SAHA/FP mediated apoptosis (eg 45 +/- 1.5% and 38.2 +/- 2.0% apoptotic vs 78 +/- 1.5% in controls) and Bid cleavage. SAHA/FP induced-apoptosis was unaffected by the free radical scavenger L-N-acetyl cysteine or the PKC inhibitor GFX. Finally, ectopic Bcl-2 expression marginally attenuated SAHA/FP-related apoptosis/cytochrome c release, and failed to restore clonogenicity in cells exposed to these agents. Together, these findings indicate that SAHA and FP interact synergistically to induce mitochondrial damage and apoptosis in human leukemia cells, and suggest that this process may also involve engagement of the caspase-8-dependent apoptotic cascade. PMID- 12094260 TI - Haplotype study of microsatellites flanking the t(15;17) breakpoint in acute promyelocytic leukemia patients from North Portugal. AB - A higher frequency of acute promyelocytic leukemia (APL) has been noted in countries of Southern Europe and among 'Latino' patients of the United States with acute myeloid leukemia (AML). In order to discover whether there is any genetic predisposition to the disease, we analyzed microsatellites flanking PML and RARalpha genes in 29 t(15;17) APL patients from North Portugal and compared them with a control group of 123 healthy individuals. Fluorescent PCR products were analyzed using an automated capillary electrophoresis system and allele and haplotype frequencies of the two populations were determined. No significant differences were found, suggesting the same genetic origin of patients and healthy individuals. As suggested by the four microsatellites screened, MSI (microsatellite instability) does not explain the increased incidence of t(15;17) APL in this Portuguese population. These results intend to be a new approach to the study of APL, reflecting the particularity of the disease. PMID- 12094259 TI - The antifungal antibiotic clotrimazole alters calcium homeostasis of leukemic lymphoblasts and induces apoptosis. AB - Clotrimazole is an antimycotic imidazole derivative that interferes with cellular Ca(2+) homeostasis. We investigated the effects of clotrimazole on acute lymphoblastic leukemia (ALL) cells. Treatment with 10 microM clotrimazole (a concentration achievable in vivo) reduced cell recovery from cultures of all nine ALL cell lines studied (B-lineage: OP-1, SUP-B15, RS4;11, NALM6, REH, and 380; T lineage: MOLT4, CCRF-CEM, and CEM-C7). After 4 days of culture, median cell recovery was 10% (range, <1% to 37%) of cell recovery in parallel untreated cultures. Clotrimazole also inhibited recovery of primary ALL cells cultured on stromal feeder layers. After leukemic cells from 16 cases of ALL were cultured for 7 days with 10 microM clotrimazole, median cell recovery was <1% (range, <1% to 16%) of that in parallel untreated cultures. Clotrimazole was active against leukemic cells with genetic abnormalities associated with poor response to therapy and against multidrug-resistant cell lines. In contrast, mature T lymphocytes and bone marrow stromal cells were not affected. Clotrimazole induced depletion of intracellular Ca(2+) stores in ALL cells, which was followed by apoptosis, as shown by annexin V binding and DNA fragmentation. Thus, clotrimazole is cytotoxic to ALL cells at concentrations achievable in vivo. PMID- 12094261 TI - Biased Iglambda expression in hypermutated IgD multiple myelomas does not result from receptor revision. AB - Normal IgM(-)IgD(+) CD38(+) B cells and IgM(-)IgD(+) multiple myelomas (MM) are characterized by Cmu deletion, biased Iglambda expression and hypermutated IgV regions. The predominant Iglambda usage has been proposed as resulting from secondary Ig gene rearrangements during extensive clonal expansion in the germinal center environment. Here, four cases of IgDlambda MM were studied to address the question of light chain receptor revision in a 'single cell' model. Detailed analyses of both IGK and IGL alleles of each case were performed by Southern blotting, (RT-) PCR, and sequencing. The expressed IgV genes were extensively mutated and Cmu deletion was confirmed in two cases. In addition, in the four MM a total of six non-functional deletional IGK rearrangements were identified, which proved to be unmutated. We conclude that IgD myelomas indeed originate from (post) germinal center B cells in which, in spite of the fact that they are hypermutated, there is no evidence of receptor revision. PMID- 12094262 TI - The cholesterol lowering drug lovastatin induces cell death in myeloma plasma cells. AB - Lovastatin is an irreversible inhibitor of HMG-CoA reductase and blocks the production of mevalonate, a critical compound in the production of cholesterol and isoprenoids. Isoprenylation of target proteins, like the GTP-binding protein Ras, is essential for their membrane localization and subsequent participation in intracellular signaling cascades. Lovastatin effectively decreased the viability of plasma cells from cell lines (n = 10) and myeloma patients' samples (n = 8) in a dose- and time-dependent way. Importantly, co-incubation of lovastatin with dexamethasone had a synergistic effect in inducing plasma cell cytotoxity. This effect was not the consequence of a change in the protein expression levels of Bcl-2 or Bax induced by lovastatin. The decrease in plasma cell viability was the result of induction of apoptosis and inhibition of proliferation. Mevalonate effectively reversed the cytotoxic and cytostatic effects of lovastatin in plasma cells. The cytotoxic activity of lovastatin was higher in Pgp expressing cell lines, but did not correlate with the multidrug resistance (MDR)-related proteins LRP, Bcl-2 and Bax. Lovastatin treatment resulted in a shift of Ras localization from the membrane to the cytosol that was reversed by mevalonate. The data presented in this paper warrant study of lovastatin alone or in combination with therapeutic drugs, in the treatment of myeloma patients. PMID- 12094263 TI - T cell receptor gamma gene rearrangements as targets for detection of minimal residual disease in acute lymphoblastic leukemia by real-time quantitative PCR analysis. AB - Several studies have shown that quantitative detection of minimal residual disease (MRD) predicts clinical outcome in childhood acute lymphoblastic leukemia (ALL). In this report we investigated the applicablility of T cell receptor gamma (TCRG) gene rearrangements as targets for MRD detection by real-time quantitative PCR analysis. Seventeen children with precursor-B-ALL and 15 children with T-ALL were included in this study. Using an allele-specific (ASO) forward primer in combination with germline Jgamma reverse primers and Jgamma TaqMan probes, a reproducible sensitivity of < or =10(-4) (defined by strict criteria) was obtained in only four out of 19 (21%) TCRG gene rearrangements in precursor-B-ALL patients and in 10 out of 15 (67%) TCRG gene rearrangements in T-ALL patients. The main reason for not obtaining a reproducible sensitivity of < or =10(-4) in approximately 60% of cases was the non-specific amplification of TCRG gene rearrangements in normal T-lymphocytes. A maximal sensitivity of < or =10(-4) (defined by less strict criteria) was obtained in 42% of TCRG gene rearrangements in precursor-B-ALL patients. The number of inserted nucleotides was significantly higher in T-ALL (mean: 8.5) as compared to precursor-B-ALL (mean: 6.8) and appeared to be the most important predictor for reaching a reproducible sensitivity < or =10(-4). The usage of a touchdown PCR or the usage of an ASO reverse primer in combination with Vgamma member forward primers and TaqMan probes did not clearly improve the overall results. Nevertheless, RQ-PCR analysis of TCRG gene rearrangements in follow-up samples obtained from 12 ALL patients showed the applicability of this method for MRD detection. We conclude that RQ PCR analysis of TCRG gene rearrangements can be used for the detection of MRD, but that sensitivities might be limited due to non-specific amplification. This method is applicable in the majority of T-ALL patients and in almost half of precursor-B-ALL patients, particularly when used as second-choice target for confirmation of the MRD results obtained via the first-choice target. PMID- 12094264 TI - Real-time quantitative PCR detection of WT1 gene expression in children with AML: prognostic significance, correlation with disease status and residual disease detection by flow cytometry. AB - The clinical significance of WT1 gene expression at diagnosis and during therapy of AML has not yet been resolved. We analysed WT1 expression at presentation in an unselected group of 47 childhood AML patients using real-time quantitative reverse-transcription PCR. We also showed that within the first 30 h following aspiration RQ-RT-PCR results were not influenced by transportation time. We observed lower levels of WT1 transcript in AML M5 (P = 0.0015); no association was found between expression levels and sex, initial leukocyte count and karyotype-based prognostic groups. There was significant correlation between very low WT1 expression at presentation and excellent outcome (EFS P = 0.0014). Combined analysis of WT1 levels, three-colour flow cytometry residual disease detection and the course of the disease in 222 samples from 28 children with AML showed remarkable correlation. Fourteen patients expressed high WT1 levels at presentation. In eight of them, who suffered relapse or did not reach complete remission, dynamics of WT1 levels clearly correlated with the disease status and residual disease by flow cytometry. We conclude that very low WT1 levels at presentation represent a good prognostic factor and that RQ-RT-PCR-based analysis of WT1 expression is a promising and rapid approach for monitoring of MRD in approximately half of paediatric AML patients. PMID- 12094265 TI - Clonal Ph-negative hematopoiesis in CML after therapy with imatinib mesylate is frequently characterized by trisomy 8. PMID- 12094266 TI - ABL-specific tyrosine kinase inhibitor imatinib as salvage therapy in a child with Philadelphia chromosome-positive acute mixed lineage leukemia (AMLL). PMID- 12094268 TI - The class II tumour suppressor gene RARRES3 maps to 11q12, not 11q23. PMID- 12094271 TI - [Surgical treatment of fractures of the clavicle--Indication, surgical technique and results]. AB - 46 cases of clavicular fracture (25 of the middle third and 21 of the distal third) are reviewed concerning indications for surgery, surgical technique and postoperative results. In 32 cases surgery was necessary due to dislocation (in 7 cases with a rupture of the coracoclavicular ligaments), 3 times due to a complex trauma of the shoulder joint, 3 times due to a local neurovascular involvement, 3 times due to a delayed fracture healing and 3 times due to a pathological fracture. In two cases an open fracture required osteosynthesis. The most frequent surgical procedure was plate osteosynthesis using the following materials: 3.5 mm LCDC plate, the so called "Balser plate" and the "Ulrich plate", 3.5 mm reconstruction plate, 3.5 mm T-plate and compound plate. In 14 cases autogenous bone grafting was performed. Additional suture of the coracoclavicular ligaments was indicated in 7 patients. In every case early physiotherapy followed the surgical procedure. COMPLICATIONS: in 2 cases an avascular pseudarthrosis emerged after plate osteosynthesis without autogenous bone grafting. In one case a non-union developed due to infection. Moreover we found a loosening of a "Balser plate" and in one patient the development of keloid tissue. The functional outcome according to the Constant score was good. 32 patients achieved 89 out of 100 possible points (average follow up 31 months). In distal clavicle fractures we prefer the use of the "Balser plate" combined with a suture of the coracoclavicular ligaments. Due to the occurrance of delayed fracture healing with pseudarthrosis (2/25) in fractures of the middle third, the indication for surgical treatment has to be discussed. In case of local soft tissue trauma and damage of the periostal blood supply, plate osteosynthesis in combination with autogenous bone grafting should be performed. PMID- 12094272 TI - [Reducing approach-associated morbidity in fracture dislocation of the femoral head--a longitudinal study (1982-2000)]. AB - Fractures of the femoral head occur during displacement injuries of the hip joint (Pipkin-type fractures). Reasons are high-energy traumas, usually dash-board injuries. Typical complications are posttraumatic necrosis of the femoral head (prevalence according to the literature: 15-66 %), and arthritis of the hip joint. It is yet uncertain, however, whether the type of surgical approach can influence the rate of necrosis. From June 1982 to December 2000, a total of 30 patients underwent surgery for Pipkin-type fractures, with 28 of them being posterior displacements, and 2 being anterior ones. Average age was 35.8 years, 2/3 were male, and 1/3 female. Total hip prosthesis was implanted primarily in 4 cases. 26 underwent osteosynthesis. Anterior displacements were stabilized via a lateral approach, whereas posterior ones were managed via a posterior approach. Reason for this procedure was the intention to use the one side of the joint capsule for approach, that had been torn already by the displacement-injury. 21 of 26 operatively stabilized patients were followed-up between 6 and 54 months postoperatively. With this regimen of treatment, we had to face no case of necrosis of the femoral head. For fracture displacement of the femoral head we therefore suggest a posterior approach in posterior displacement, as well as an anterior approach for anterior displacement. Using this principle, rate of necrosis of the femoral head may clearly diminish (in our series 0 out of 21). PMID- 12094273 TI - [Combined posterior and anterior endoscopic supported instrumentation of the thoracic and lumbar spine. Arguments, concepts and first clinical results]. AB - The operative therapeutic principles of thoracic and lumbar spine injuries are based on reposition, decompression and stabilization of the unstable area. Secondary loss of correction and consecutive deformation have negative impact on the long-term results after defect fractures of vertebral bodies and will be minimized only by the reconstruction of all involved spinal columns. With dissemination of thoracoscopic techniques at the thoracic spine and minimal invasive retroperitoneal approaches at the lumbar spine a decrease of the morbidity was achieved with equal effectivity for recalibration and fusion. Our experiences with the first consecutive 42 patients, treated minimal invasive are presented. Indications for anterior thoracoscopic and minimal invasive instrumentations after posterior transfixation are remaining osseous defects of the end plates of more than a quarter of the volume of the involved vertebra in case of migration of the vertebral disc, wedging of the vertebral body after posterior reposition of more than 10 degrees and persisting anterior encroachment of the spinal canal of more than 30 %. The decision is based on radiographs and CT-scans, performed after posterior stabilization. PMID- 12094275 TI - [Hip replacement after failed internal fixation in patients with proximal femur fracture--a simple procedure?]. AB - Between January 1995 and December 2000 we operated on 1 437 patients with fractures of the proximal femur. In 789 cases an osteosynthesis was performed and in 648 cases a prosthesis was implanted. In the group of patients with primary osteosynthesis we stabilized 89 fractures with screws, in 319 patients a dynamic hip screw (DHS) and in 381 patients a Proximal-Femurnail (PFN) were implanted. Because of major complications we had to reoperate on 42 patients (5.3 %) with primary osteosynthesis. For analysis of this high complication rate we studied the fracture type, the type of complication, the operation time, the primary and secondary implant type and the time of postoperative mobilisation in this group of 42 patients. We saw 27 type A and 15 type B fractures, according to the AO classification. In the patient group with complications 4 times a primary screw stabilisation was done, 22 dynamic hipscrews (DHS) and 16 proximal femurnails (PFN) were implanted. After changing the primary osteosynthesis into 15 hemiprostheses and 26 total prostheses of the hip, we found in over 30 % major complications. A change of the procedure after primary osteosynthesis has to be regarded as a more difficult operation than the primary implantation of a prosthesis. PMID- 12094274 TI - [Results after resection of juxta-facet-cysts - Which role does a segmental instability play?]. AB - This study demonstrates the results after operative treatment of patients suffering from a lumbar Juxta-Facet-Cyst. We point out diagnostical aspects, possible concomitant problems and deriving therapeutical consequences. Between 01. 01. 1998 and 31. 03. 2001 9 patients were operated on a synovial cyst or a ganglion of the facet joint at our department. 5 patients were female, 4 patients male with a mean age of 61 (45-70) years. The average clinical and radiological follow up was 11 (5-18) months postoperatively. The clinical examination revealed in 5 out of 9 patients a sensible deficit, in 3 out of 9 patients motoric disturbances. There was no positive sign of Lasegue. The resection of the cyst or ganglion was performed in all cases via a dorsal approach. 8 patients underwent for a spinal stenosis and/or an existing instability a laminectomy and a spondylodesis with an internal fixateur. The histological findings showed a synovial cyst in 6 cases and a ganglion cyst in 3 cases. At the follow up all preoperative sensible or motorical deficits had resolved. Juxta-Facet-Cysts are discovered in some cases intraoperatively by chance, the radiological methods (CT, MRT) are especially in case of a concomitant spinal stenosis uncertain. During all operations, which are performed for a spinal stenosis, one should look for a cyst. The resection of the cyst leads to good functional results, a possible instability should by addressed by a spondylodesis. PMID- 12094276 TI - [Evaluation and results of the gliding nail in the elderly patient]. AB - Evaluation of the gliding nail in the elderly patient with unstable fracture of the proximal femur. Primary aims of the study were the description of surgical procedure, the specific complications, postoperative pain, recovered mobility and social status after nailing procedure. The observation period of this prospective non-randomized observational cohort study was from 10/97 to 07/00. 121 patients (90 female, 31 male) with unstable femoral fracture were included. Data evaluation included type of fracture, surgical data, procedure specific complications and pre and post surgical development (pain, mobility and social status). The evaluation of the outcome was performed at least 3 months (median 10.4 months) after surgical intervention by telephone-interview. A subgroup of 20 patients was re-examined. 24 A1-, 65 A2-, 16 A3-, 11 subtrochanteric and 5 other fractures were stabilized with a gliding nail. The mean time of surgery was 77 minutes. In 15 out of 121 patients complications lead to a second surgical intervention. In the follow-up examination we were able to obtain following findings: 33.9 % vs. 56.2 % (preoperative) of patients were self-contained. The need for professional care was unchanged. 23.5 % vs. 58.7 % (preoperative) showed unlimited mobility. 20.3 % vs. 12.8 % complained light or moderate pain. According to our experience the gliding nail is a safe and minimal invasive system with a low complication rate. In consideration of patient based outcome the gliding nail shows good results compared to other treatment options. For the given reasons the authors prefer the gliding nail as treatment of choice for proximal femoral fractures. PMID- 12094277 TI - [The femoral neck fracture in the elderly patient - cemented or cementless hip arthroplasty?]. AB - This is a report on the treatment results of femoral neck fractures with a cemented (group 1) and cementless (group 2) type of hip prosthesis, resp. 72 patients were enrolled. 35 patients were treated with a cemented hip prosthesis (mean age: 78 years), and 37 patients with a cementless modular hip prosthesis (mean age: 77 years). In the cemented group we observed 5 cases of hypotension during insertion of the prosthesis in the femoral shaft. One of these patients required mechanical resuscitation during surgery. In the second group 3 cases of proximal femur fissure and one case of distal femoral fracture were recorded. One year after surgery 43 patients presented for follow up evaluation (cemented group: n = 24; cementless group: n = 19). Both groups revealed comparable results according to the Harris Hip Score (75 versus 78,3 points). No prosthesis loosening was observed in either group. In our view the cemented hip prosthesis is the treatment of choice for femoral neck fractures among the old and very old, if no stabile osteosynthesis can be performed. Patients with cardiopulmonary risk factors, however, may profit from cementless hip arthroplasty to avoid the well known cardiodepression during surgery. PMID- 12094278 TI - [Achilles tendon rupture: early dynamic mobilisation after surgery]. AB - PURPOSE OF THE STUDY: It should be investigated whether the results of operative treatment of Achilles tendon ruptures can be improved by the use of a lower leg splint. PATIENTS AND METHODS: We report on 58 patients with a subcutaneous Achilles tendon rupture, who underwent a standardised suture repair (Bunnell suture) followed by an early functional dynamic treatment with a newly developed lower leg splint. RESULTS: 45 patients, 3 women and 42 men, were examined on average 30 months after the operation. The cause of the rupture was predominantly a sporting activity. There were no postoperative complications, no rerupture and no infection. The ultrasonic investigation showed a very good gliding ability of the operated tendons, in the compartment I with 38 and in the compartment II with 7 of the 45 patients. According to the Trillat classification 27 patients scored excellent and 18 patients good. CONCLUSION: Similar to the treatment concept after surgical repair of hand tendons an early better result can be obtained by a functional dynamic treatment regimen also in operated Achilles tendon ruptures. PMID- 12094279 TI - [Risks and injuries of go-karting]. AB - The demands on leisuretime activities are getting more exacting. So trendy sports just like inline-skating, mountain-biking, paragliding, bungee-jumping or driving go karts become increasing popular. On the other side risks and injuries of this fun sports are not enough represented. We observed in a 16 months period 5 serious accidents with go karts. The injuries included bruise of lower leg, a contusion of the stomach with a pancreatic lesion, an instable fracture of the spinal column, a difficult fracture of the knee and upper tibia and a large sized burnwound. The mean hospitalization time was 32,5 days (17-54 days) and the average age of the patients 27,7 years (9-45 years). The lack of safety devices and the subjective inexperience make go kart driving a dangerous fun. The TUV worked out guidelines for security of indoor kart centres but they don't have any legal liability in case of injury. Therefore everyone carrying out fun sports like go kart driving should be aware of the dangers and should gather information whether the accident insurance will help out in case of injury. PMID- 12094280 TI - [Decubital ulcer--causes, surgical therapy and prognosis]. AB - 7.6 % (n = 134/1 769) of all inpatients in Leuven, but 30.1 % of risk patients suffer of pressure sores (Norton Scale 10.4 points). Predisposed are geriatric patients or those of intensive care units. While fasciocutaneous flap likewise the superior gluteal artery perforator flap are more resistant to mechanical forces, myocutaneous flaps are the working horse for pressure sores with deep tissue pockets or osteitis (Campbell stage > IV). Partial resection of the median sacral crest or ischiectomy are obligatory. In analyzing a 16 years period only 33 percent of myocutaneous flaps healed uneventful, while due to multiple enumerations more than 50 percent presented complications. Partial flap and complete necrosis happened in 6 respectively in 2 percent. Rectus abdominis myocutaneous flap reconstruction as well as the free tissue transfer are useful alternatives to hip disarticulation with total thigh flap coverage or a complete fillet flap of the lower extremity. Independently of the selected closure 50 percent of all decubitus patients will develop recurrences in longterm follow up. Thus prevention is of utmost importance. In Europe the costs for the hospitalisation of a patient with concomitant decubital ulcers are 2.5-times higher than for an average, non-afflicted patient. PMID- 12094281 TI - [The importance of laparoscopy in blunt abdominal trauma]. AB - The importance of laparoscopy in the management of blunt abdominal trauma should be evaluated. Therefore we retrospectively analysed all patients with blunt abdominal trauma treated in the Department of Surgery at the Carl-Thiem-Hospital Cottbus between 1998 and 2000. Within this period a total number of 53 patients with blunt abdominal trauma underwent operative treatment, 20 (37.7 %) of them had primary laparoscopy. Of the 11 cases where laparoscopic operation could be completed without conversion to exploratory laparotomy, 8 patients had intra abdominal injuries and underwent sufficient laparoscopic treatment. The percentage of so called "negative" exploratory laparotomies within this study was 13.2 %. Our analysis suggests that laparoscopy should become firmly established in the diagnostic management and, if indicated, in the treatment of blunt abdominal trauma as well. PMID- 12094282 TI - [Visceral artery aneurysms]. AB - The purpose of this article is to review the etiology, clinical presentation and treatment options of visceral artery aneurysms (VAA) on the basis of our experiences. Visceral artery aneurysms are uncommon lesions with a frequency of 0,1-0,2 % in autopsy statistics. In fact many visceral artery aneurysms still present first with a rupture (22 %) and despite emergency laparotomies the mortality rate is about 8,5 %. The course of disease often is disastrous due to rupture of the aneurysms or thromboembolic complications, emphasizing the importance to be aware of this differential diagnosis of abdominal pain. This article covers 9 patients with VAA. 3 patients each revealed an a. lienalis aneurysm and a. gastrica aneurysm, resp. In the other 3 patients an aneurysm of the a. gastroepiploica, the a. pancreatico-duodenalis and the a. mesenterica superior resp. was proven. In 8 of 9 patients a surgical therapy of the VAA took place. Only 2 patients (22 %) were interventionally treated. 1 patient deceased due to postoperative hemorrhage. Both the surgical and the radiological intervention therapy are available for treatment of the VAA. The decision on the choice of the therapeutic procedure should be made on an individual basis. PMID- 12094283 TI - [Laparoscopic splenectomy by the "Essen-manoeuvre" for treatment of benign and malignant haematological diseases]. AB - Despite its early description, laparoscopic splenectomy has not yet reached the level of a standard operation as a therapeutic option for haematological disease, especially for malignant disorders. We performed laparoscopic splenectomies in a modified 4 port technique and dissection of the splenic vessels by the "Essen Manoeuvre" and report on 68 attempted laparoscopic splenectomies for benign (n = 42) and malignant (n = 26) haematological disorders. Conversion rate was 9 %, 30 days-mortality was 1.4 %, perioperative morbidity was 11 %, respectively. Accessory spleens were found and resected in 17 % of our patients. Laparoscopic splenectomy is a new minimally invasive option for patients with benign and malignant haematological disease fraught with a special risk of intraoperative bleeding. PMID- 12094284 TI - [Lymphadenectomy in gastric carcinoma?]. AB - Lymphadenectomy in gastric carcinoma is still discussed. Based on non-randomised and randomised studies D2-lymphadenectomy is indicated only in R0-resections. D2 lymph-node dissection requires great experience with this technique as well as for the management of possible postoperative complications. Survival advantages can be expected in patients with a limited nodal involvement, a favorable lymph node ratio between resected and involved nodes and in stage II disease. Therefore, survival improvement can be expected in 15-20 % of all R0-resections. Despite these limited prognostic advantages D2-lymphadenectomy should be performed in all R0-procedures since only the pathologic data of the resected specimen supply reliable prognostic parameter. If in the future sentinel lymphadenectomy allows selection criteria for a balanced indication for D2 lymphadenectomy in gastric carcinoma, needs to be awaited. PMID- 12094286 TI - [The iatrogenic small bowel perforation as a complication of laparoscopic cholecystectomy]. AB - Having done 1 125 laparoscopic cholecystectomies we want to report an exceptional case of perioperative bowel lesion. The symptoms began with abdominal pain on the third postoperative day, followed by an evacuation of intestinal contents out of the Robinson-Drainage. The laparotomy showed a 4 mm lesion of the terminal ileum, which could be excised and closed. Any other complications were not recognized. The reason that caused the perforation could not be clarified. Possible mechanisms will be discussed. PMID- 12094287 TI - Effect of caffeic acid on apical transporters' dysfunction of renal proximal tubule cells under oxidative stress in vitro. AB - The protective effect of caffeic acid (CA) against oxidative stress-induced inhibition of proximal tubule apical transporter was investigated. In the present study, 10 (-4) M H2O2 did not affect cell viability regardless of incubation time. However, it decreased apical transporters' activity such as Na (+)/glucose cotransporter, Na (+)/Pi cotransporter, and Na (+)/H(+) antiporter in the proximal tubule cells. CA (>10(-6) M) prevented H2O2-induced inhibition of apical transporters. Thus, we investigated its action mechanism. CA also prevented H2O2 induced lipid peroxides formation, arachidonic acid (AA) release, and Ca(2+) uptake. In conclusion, CA, in part, prevented H2O2-induced inhibition of apical transporter activity via decrease of AA release and Ca(2+) uptake in primary cultured renal proximal tubule cells. PMID- 12094288 TI - Vascular effects of a soy leaves (Glycine max) extract and kaempferol glycosides in isolated rat carotid arteries. AB - We have recently purified genistin, and six kaempferol glycosides from a soy leaves ( Glycine max L. Merr.) butanol extract. Here we report the vascular effects of the extract and purified genistin and kaempferol glycosides on contractions induced by different constricting agonists in isolated rat carotid arteries. The butanol extract relaxed artery rings preconstricted by 9,11-dideoxy 11alpha,9alpha-epoxy-methanoprostaglandin F 2 alpha (U46619) or [5 Z,9alpha,11alpha,13 E,15 S]-9,11,15-trihydroxyprosta-5,13-dienoic acid (PGF 2 alpha ) in a dose-dependent manner and this effect was independent of the presence of endothelium. The extract also inhibited the concentration-dependent contraction to U46619 with a slight reduction of the maximal response. The extract produced partial relaxation of both phenylephrine-preconstricted endothelium-intact and -denuded rings. In contrast, the extract had no effect on the contractile response to 50 mM extracellular K (+). None of the six kaempferol glycosides affected vessel tension induced by U46619. A mixture of kaempferol glycosides prepared according to their relative composition in the extract had no effect either. However, kaempferol relaxed U46619- and high K (+)-contracted rings to the same extent. Endothelium played no role in kaempferol-induced relaxation. Genistein induced concentration-dependent relaxation and this effect was attenuated in the endothelium-denuded rings. Genistin caused a smaller relaxant effect. The present results indicate that a butanol extract from soy leaves causes endothelium-independent relaxation in rat carotid artery rings. Kaempferol glycosides, accounting for approximately 48 % of the extract in weight, are not the ingredients responsible for the extract-induced relaxation. Genistein and genistin also caused relaxation, however, the dose range is beyond that of the extract causing relaxation. PMID- 12094289 TI - Inhibitory effects of glycosides from the leaves of Melaleuca quinquenervia on vascular contraction of rats. AB - Two new glycosides, 3-hydroxy-5-methoxy-4-methylphenyl beta-D-glucopyranoside (1) and 4-benzoyl-2-C-beta-glucopyranosyl-3,5-dihydroxy-6-methylphenyl beta-D glucopyranoside (2), together with four known glycosides, 2-endo-beta-D glucopyranosyloxy-1,8-cineole (3a), 2-exo-beta-D-glucopyranosyloxy-1,8-cineole (3b), roseoside (4), and citroside A (5), were isolated from the methanolic extract of leaves of Melaleuca quinquenervia. Their structures were elucidated on the basis of spectroscopic analysis. Compounds 1, 2a and 3 inhibited contractile response induced by phenylephrine in aortic rings from Sprague-Dawley rats. This inhibition was independent of the endothelium. Compounds 2 and 4 significantly relaxed precontracted aortic rings, in an endothelium-dependent manner. Pretreatment of N(omega)-nitro-L-arginine (L-NNA), a nitric oxide synthase inhibitor, partially attenuated the vasorelaxation induced by both compounds, suggesting that nitric oxide was likely the responsible mediator. The rank-order potency (EC 50 value) of vasorelaxing activities of these compounds is 4 > 2 > 2a > 3 > 1. PMID- 12094290 TI - Ginsenosides from Panax ginseng differentially regulate lymphocyte proliferation. AB - We have examined the immunosuppressive effects of representative ginsenosides (Rb1, Rb2, Re and Rg1) from Panax ginseng C. A. Meyer on CD4+ and CD8+ lymphocyte proliferation. Ginsenosides differentially modulated lymphocyte proliferation induced by concanavalin A (Con A), lipopolysaccharide (LPS), phytohemaglutinin (PHA) and interleukin-2 (IL-2). Thus, Rb1 and Re significantly enhanced Con A induced lymphocyte proliferation, whereas Rg1 did not affect the proliferation. Interestingly, however, Rb2 strongly blocked Con A, LPS and PHA-induced lymphocyte proliferation with the IC50 values of 21.8, 29.0 and 24.0 microM, respectively. Moreover, Rb2 inhibited Con A-stimulated IL-2 production with an IC 50 of 13.3 microM. In the IL-2-stimulated CD8+ T cell (CTLL-2) proliferation assay, Re and Rg1 showed strong suppressive effects with IC50 values of 57.5 and 64.7 microM, respectively. In contrast, neither Rb1 nor Rb2 did inhibit CTLL-2 cell proliferation at tested concentrations. These results suggest that ginsenosides from P. ginseng may modulate lymphocyte proliferation in a different manner. PMID- 12094291 TI - Inhibition of intracellular calcium elevation and blunting of vasopressor response due to serotonin by ginkgolide B. AB - The vascular effects of each individual composition in the extract of Ginkgo biloba have not been clarified. In this work, we have investigated whether ginkgolide B (GKB), a terpene lactone component from Ginkgo biloba, modulates intracellular calcium ([Ca (2+)] i) of vascular smooth muscle cells (smc) and how it influences the vasopressor response in vitro caused by serotonin (5-HT). GKB (3.2 - 9.6 microM) selectively decreased serotonin-induced [Ca (2+)] i elevations in cultured smc from bovine aorta in dose-dependent manner, while it had no effects on both resting [Ca (2+)] i and potassium-elicited [Ca (2+)] i elevations. On the other hand, GKB in a concentration of 3.2 - 9.6 microM moderately decreased the maximal pressor efficacy of serotonin by 14 - 45 % in rat mesenteric vascular beds, and the EC50 remained unchanged. The comparison study also showed GKB had no effects on noradrenaline-elicited pressor reactions. These results implicate that GKB may selectively inhibit serotonin-mediated [Ca (2+)] i mobilization in vascular smc and non-competitively alleviate the vasopressor effect of serotonin in vitro. PMID- 12094292 TI - Inhibition of cultured bovine aortic endothelial cell proliferation by sodium spirulan, a new sulfated polysaccharide isolated from Spirulina platensis. AB - Sodium spirulan (Na-SP) is a sulfated polysaccharide isolated from the blue-green alga Spirulina platensis, which consists of two types of disaccharide repeating units, O-hexuronosyl-rhamnose (aldobiuronic acid) and O-rhamnosyl-3-O methylrhamnose (acofriose) with sulfate groups, other minor saccharides and sodium ion. Vascular endothelial cells are present on the inner surface of blood vessels in a monolayer and have anticoagulant properties. To address the question whether Na-SP influences the maintenance of endothelial cell monolayers, we investigated the proliferation of cultured bovine aortic endothelial cells treated with Na-SP. It was found that Na-SP has an inhibitory activity on endothelial cell proliferation accompanied with suppression of whole protein synthesis but without non-specific cell damage. The inhibitory activity of Na-SP was the strongest when compared to that of heparan sulfate, heparin, dextran sulfate, dermatan sulfate, chondroitin sulfate A/C and hyaluronan. Furthermore, it was shown that the inhibitory activity of Na-SP disappeared by either desulfation or depolymerization. The present data suggest that Na-SP is a unique sulfated polysaccharide that strongly inhibits vascular endothelial cell proliferation, and the inhibitory activity requires polymerization of sulfated O rhamnosyl-acofriose repeating units. PMID- 12094293 TI - Hepatoprotective and therapeutic effects of tetramethylpyrazine on acute econazole-induced liver injury. AB - 2,3,5,6-Tetramethylpyrazine (TMP) is well known as a true calcium antagonist. The aim of this study was to investigate the hepatoprotective and therapeutic effects of TMP on acute econazole-induced liver injury. The hepatological effect of various concentrations of TMP was first assessed by the biochemical assays of SGOT and SGPT and then by hepatohistological microscopic examination. The dose response relationship of liver injury induced by various doses of econazole was observed simultaneously from serum biochemical assay of SGOT and SGPT, and also from hepatohistological microscopic examination, by determination of the hepatoprotective effects of various concentrations of TMP on SGOT and SGPT elevation induced by a hepatotoxic dose of econazole (300 mg/kg). The inhibitory effect of various concentrations of TMP or vitamin E (positive control, 0.5 mM in vitro, 0.69 mM in vivo) on FeCl 2 -induced (in vitro) or econazole-induced (in vivo) lipid peroxidation was also investigated. The superoxide scavenging activity of various concentrations of TMP in econazole-damaged rat liver homogenate was assessed by the cytochrome C reduction method. Results showed that the hepatoprotective effect of TMP might be, at least in part, due to its inhibitory ability on membrane lipid peroxidation and free radical formation, or due to its free radical scavenging ability. Improvement of serum transaminases and MDA levels in rat liver homogenate, hepatohistological microscopic examination, and assessment of free radical scavenging activity by the cytochrome C reduction method were used to detect hepatoprotective and therapeutic effects of TMP on acute econazole-induced liver injury. PMID- 12094295 TI - Flavonoids with activity against methicillin-resistant Staphylococcus aureus from Dalea scandens var. paucifolia. AB - Bioassay-guided fractionation of the ethyl acetate extract of the roots of Dalea scandens (Miller) R. Clausen var. paucifolia led to the isolation of new flavonoids, 2( S)-5'-(-1"',1"'-dimethylallyl)-8-(3",3"-dimethylallyl)-2',4',5,7 tetrahydroxyflavanone, 2( S)-5'-(1"',1"'-dimethylallyl)-8-(3",3"-dimethylallyl) 2'-methoxy-4',5,7-trihydroxyflavanone and 5'-(1"',1"'-dimethylallyl)-8-(3",3" dimethylallyl)-2',4',5,7-tetrahydroxyflavone. Structure elucidation was carried out by spectroscopic methods. All three compounds showed significant activity against both methicillin-susceptible and methicillin-resistant Staphylococcus aureus. PMID- 12094294 TI - Antiulcerogenic activity of some sesquiterpene lactones isolated from Artemisia annua. AB - Artemisinin 1, dihydro-epideoxyarteannuin B 2 and deoxyartemisinin 3 were isolated from the sequiterpene lactone-enriched fraction obtained from the crude ethanolic extract of Artemisia annua L. These compounds were tested on ethanol and indomethacin-induced ulcer models. Compound 1 did not afford cytoprotection under the experimental models tested. Only compounds 2 and 3 decreased the ulcerative lesion index produced by ethanol and indomethacin in rats. These compounds did not demonstrate antiulcerogenic activity when tested on the ethanol induced ulcer model, with previous administration of indomethacin, suggesting that the antiulcerogenic activity is a consequence of prostaglandin synthesis increase. PMID- 12094296 TI - Pharmacologically active ellagitannins from Terminalia myriocarpa. AB - A new ellagitannin, methyl (S)-flavogallonate (14) along with fourteen known compounds, gallic acid, methyl gallate, ethyl gallate, 2,3-di-O-[( S) 4,5,6,4',5',6'-hexahydroxybiphenyl-2,2'-diyldicarbonyl]-(alpha/beta)-D glucopyranose (4), vitexin, isovitexin, orientin, iso-orientin, kaempferol 3-O beta-D-rutinoside, rutin, neosaponarin, ellagic acid, flavogallonic acid (13), and (alpha/beta)-punicalagin (15) have been isolated from the leaves of Terminalia myriocarpa Heurck. Protective effect of the major and structurally related compounds 4, 13, 15 and the new compound 14 against CCl 4 -induced hepatotoxicity has been evaluated and compared, using adult male rats weighing 200-250 g. Serum levels of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), lipid peroxide and nitric oxide production were significantly increased by administration of CCl 4 to rats and then reduced significantly only by treatment with compounds 4, 14 and 15 in a dose-dependent manner. Comparison of the protective properties of these compounds showed that compound 14 is more potent than compound 15 than 4 and that compound 13 has a non significant effect at the used two dose levels. PMID- 12094297 TI - Cytotoxic ent-kaurane diterpenoids from Isodon eriocalyx var. laxiflora. AB - Three new ent-kaurane diterpenoids laxiflorin E (1), laxiflorin H (6) and laxiflorin I (8) were isolated from the leaves of Isodon eriocalyx var. laxiflora, along with nine known diterpenoids, eriocalyxin A (2), laxiflorin C (3), laxiflorin D (4), laxiflorin A (5), maoecrystal S (7), maoecrystal Q (9), eriocalyxin B (10), maoecrystal C (11) and enmelol (12). The structures of the new compounds were determined by spectroscopic methods. Compounds 1-5 are 6,7 seco-ent-kaurane-7,20-olide diterpenoids, and 6-12 belong to 7,20-epoxy-ent kauranoids. The spectral data of enmelol (12) are reported here for the first time. Ten of these compounds were tested for their cytotoxicity toward K562 and T24 human tumor cells. Compounds 1, 3 and 10 showed significant inhibitory effects on K562 cells with IC50 values 0.077, 0.569 and 0.373 microg/mL, and compounds 1 and 10 also demonstrated significant inhibitory activities toward T24 cells with IC50 values 0.709 and 0.087 microg/mL. Compounds 8 and 11 also displayed inhibitory effect on both two kinds of cells with IC 50 value less than 6.5 microg/mL. PMID- 12094298 TI - Alkaloids, amides and antispasmodic activity of Zanthoxylum hyemale. AB - Two quinoline alkaloids, (-)-R-geilbalansine (1) and hyemaline (2), as well as aromatic amide, N-[2-(3,4-dimethoxyphenyl)-2-methoxyethyl)-2 methoxyethyl]benzamide (O-methylbalsamide) (3), were isolated as new natural products from the stem barks of Zanthoxylum hyemale, together with seven known compounds. Their structures were determined on the basis of spectroscopic data (IR, (1)H- and (13)C-NMR, MS). In addition, the antispasmodic activity of the crude extract of Z. hyemale and three other more abundant isolated compounds (4, 5 and 10) were studied in two different antispasmodic test models on isolated rat ileum and only the crude ethanolic extract presented antispasmodic activity. PMID- 12094299 TI - Antiviral and antioxidant activity of flavonoids and proanthocyanidins from Crataegus sinaica. AB - The antiviral and antioxidant activity of some fractions and of a series of flavonoids and proanthocyanidins obtained from Crataegus sinaica (Rosaceae) was evaluated. The O-glycosidic flavonoids and the oligomeric proanthocyanidins exhibited significant inhibitory activity against herpes simplex virus type 1 (HSV-1), which was shown to be due to an extracellular mechanism for procyanidin C-1. Procyanidin C-1 also had the highest antioxidant activity in both the microsomal lipid peroxidation and the hydroxyl radical scavenging assay. In addition to the previously reported phenolic compounds, the pentacyclic triterpenoid ursolic acid (1) and a tetrameric (2) and pentameric procyanidin (3) are reported for the first time. PMID- 12094300 TI - Antibacterial activity of calozeyloxanthone isolated from Calophyllum species against vancomycin-resistant Enterococci (VRE) and synergism with antibiotics. AB - Calozeyloxanthone ( 1) was re-isolated from the root bark of Calophyllum moonii, an endemic species of Sri Lanka, and found to be active against vancomycin resistant Enterococci (VRE) and vancomycin-sensitive Enterococci (VSE) with MIC values of 6.25 microg/ml and 12.5 microg/ml, respectively. Further, a marked synergism between 1 and vancomycin hydrochloride (VCM) against VRE was also observed. These findings suggest that 1 in combination with VCM against VRE may be useful in controlling VRE infections. PMID- 12094301 TI - In vitro antiplasmodial activity of Tithonia diversifolia and identification of its main active constituent: tagitinin C. AB - The antimalarial properties of Tithonia diversifolia, an Asteraceae traditionally used to treat malaria, were investigated in vitro against three strains of Plasmodium falciparum. The ether extract from aerial parts of the plant collected in Sao Tome e Principe, demonstrated good antiplasmodial activity (IC 50 on FCA strain: 0.75 microg/ml). A bioassay guided fractionation of this extract led to the isolation of the known sesquiterpene lactone tagitinin C as an active component against Plasmodium (IC 50 on FCA strain: 0.33 microg/ml), but also possessing cytotoxic properties (IC 50 on HTC-116 cells: 0.706 microg/ml). PMID- 12094302 TI - Inhibitory effects of natural sesquiterpenoids isolated from the rhizomes of Curcuma zedoaria on prostaglandin E2 and nitric oxide production. AB - Beta-turmerone and ar-turmerone, sesquiterpenoids isolated from the rhizome of Curcuma zedoaria, inhibited lipopolysaccharide (LPS)-induced prostaglandin E 2 production in cultured mouse macrophage cell RAW 264.7 in a dose-dependent manner (IC 50 = 7.3 microM for beta-turmerone; IC 50 = 24.0 microM for ar-turmerone). In addition, these compounds exhibited inhibitory effects on LPS-induced nitric oxide production in the cell system. PMID- 12094303 TI - Possible artefacts in the in vitro determination of antimalarial activity of natural products that incorporate into lipid bilayer: apparent antiplasmodial activity of dehydroabietinol, a constituent of Hyptis suaveolens. AB - Dehydroabietinol isolated from Hyptis suaveolens (L.) Poit. was found to inhibit growth of chloroquine-sensitive as well as chloroquine-resistant strains of Plasmodium falciparum cultivated in erythrocytes in vitro (IC 50 26-27 microM). However, erythrocytes exposed to dehydroabietinol were transformed in a dose dependent manner towards spherostomatocytic forms with concomitant formation of endovesicles, as disclosed by transmission electron microscopy. The erythrocyte shape alterations caused by dehydroabietinol correlated well with its apparent IC 50 value. Thus, dehydroabietinol incorporates into the erythrocyte membrane, and since invasion and survival of Plasmodium parasites is known to depend on the function of the erythrocyte membrane, the observed antiplasmodial effect of dehydroabietinol is presumably an indirect effect on the host cell. Because of these findings, microscopic investigations should be generally used to support claims of antimalarial effects of apolar natural products. PMID- 12094304 TI - Apoptosis of human highly metastatic lung cancer cell line 95-D induced by acutiaporberine, a novel bisalkaloid derived from Thalictrum acutifolium. AB - The present study demonstrates for the first time that acutiaporberine, a bisalkaloid isolated from the tranditional Chinese medical herb Thalictrum acutifolium (Hand.-Mazz.) Boivin. (TAB), induces apoptosis of a cultured highly metastatic human lung cancer cell line 95-D. Immunohistochemistry assay (IHC) and Western-blot analysis show down-regulation of bcl-2 gene and up-regulation of bax gene and c-myc gene in the treated cells. These results suggest that acutiaporberine may be a natural potential apoptosis-inducing agent for highly metastatic lung cancer. PMID- 12094305 TI - Cytotoxic constituents from Aquilegia ecalcarata. AB - A new alkaloid, 7-hydroxy-4-(5'-hydroxymethylfuran-2'-yl)-2-quinolone (1), and a new nitrile derivative, 3alpha,4beta-dihydroxy-6-oxo-1-cyclohexene-1-acetonitrile (2), together with three known oxoaporphine alkaloids, were isolated from the whole plant of Aquilegia ecalcarata. Their structures were established on the basis of spectral evidence. Their in vitro cytotoxicity against different classes of cancer cell lines, including GLC-82 and HCT were determined. The new alkaloid 1 [IC 50 (GLC-82) 8.8 +/- 0.2 microM, IC 50 (HCT) 10.1 +/- 0.3 microM] and hernandonine ( 3) [IC 50 (GLC-82) 7.6 +/- 0.5 microM, IC 50 (HCT) 8.2 +/- 0.5 microM] exhibited cytotoxicity towards the cancer cell lines. PMID- 12094306 TI - Cytotoxic anthraquinone derivatives from Picramnia antidesma. AB - Activity-guided investigation of crude extracts prepared from the root bark of Picramnia antidesma, a medicinal plant long used for the treatment of malaria in tropical areas of the Americas, when tested on KB cells led to the isolation of a new compound, 10-epi-uveoside, from a cytotoxic fraction containing a rich mixture of anthrone glycosides. The antiplasmodial activity proved to be a result of the high levels of cytotoxicity displayed by the anthraquinone derivatives and therefore infusions from this crude drug lack the selectivity index needed to be an effective antimalarial agent. PMID- 12094307 TI - Aceriphyllic acid A, A new ACAT inhibitory triterpenoid, from Aceriphyllum rossii. AB - Two new triterpenoids along with three known ones, 3-oxoolean-12-en-27-oic acid (1), 3alpha-hydroxyolean-12-en-27-oic acid (2) and 3beta-hydroxyolean-12-en-27 oic acid (3), were isolated from Aceriphyllum rossii. The structures of the new compounds were determined to be a 3alpha,23-dihydroxyolean-12-en-27-oic acid (4) and a 3alpha,23-dihydroxyolean-12-en-29-oic acid (5) by spectroscopic and chemical methods; they were designated aceriphyllic acids A and B, respectively. Compounds 2, 3 and 4 remarkably inhibited the activity of ACAT. PMID- 12094308 TI - Phagnalon rupestre as a source of compounds active on contact hypersensitivity. AB - The effect of Phagnalon rupestre MeOH extract on dinitrofluorobenzene- and sheep red blood cells-induced hypersensitivity was investigated. Eight compounds were identified: three dimethylallyl-hydroquinone glucosides (1 - 3), 3,5- and 4,5-di O-caffeoylquinic acid methyl esters (4 and 5), their free carboxyl analogues (6 and 7), and luteolin 7-O-beta-glucoside (8). All were tested for dinitrofluorobenzene-induced contact hypersensitivity inhibitory activity. Flavonoid 8 was the most active (49 % and 79 % inhibition at 24 and 96 h, respectively). The hydroquinones 1, 2 and 3 were effective at 96 h after challenge (62 %, 73 % and 60 % inhibition, respectively), while some of the dicaffeoylquinic derivatives (4 and 7) produced slightly lower reduction of the inflammatory reaction. PMID- 12094309 TI - Ferulagone: a new monoterpene ester from Ferulago thirkeana essential oil. AB - Essential oil obtained by micro-distillation and hydrodistillation of the endemic Ferulago thirkeana (Boiss.) Boiss. (Apiaceae) was analysed by GC/MS. An unknown component isolated from the essential oil was characterized by chromato-spectral techniques (1D-, 2D-NMR, HRMS, IR and UV) as (1 S)-2,6,6-trimethyl-4 oxobicyclo[3.1.1]hept-2-enyl(2 E)-2-methylbut-2-enoate (= ferulagone or 1 angeloyloxyverbenone) (1). Biological activities of the essential oil and the purified major component (1) were tested against various human pathogenic microorganisms resulting in moderate inhibition (62.5 - 125 microg/ml). PMID- 12094310 TI - A new diterpene from Tanaecium jaroba. AB - One new diterpene, 2 alpha-hydroxy-12beta-hydroxy-isopimara-8(14), 15-diene, and six known compounds as triterpenes, sterols and fatty acid, were isolated from the stem bark of Tanaecium jaroba (Bignoniaceae), a Bolivian plant used in traditional medicine. Their structures were established mainly by 1D and 2D NMR (COSY, HMQC, HMBC, ROESY) and their antiplasmodial activities were evaluated in vitro against Plasmodium falciparum. PMID- 12094311 TI - Quantitative determination of valepotriates from Valeriana native to South Brazil. AB - Valtrate, DIA-valtrate, acevaltrate, 1-beta-acevaltrate and didrovaltrate have been quantitatively estimated by reversed-phase HPLC in the leaves, flowers, stems and roots of Valeriana glechomifolia Meyer, V. catharinensis Graebn., V. chamaedryfolia Cham. & Schltdl., V. eichleriana (C.A.Mull.) Graebn., V. polysthachya Smith, V. scandens L., V. eupatoria Sobral, V. salicariifolia Vahl and V. tajuvensis Sobral. All plants presented valepotriates being V. glechomifolia the richest one, followed by V. eupatoria, V. eichleriana and V. tajuvensis. PMID- 12094312 TI - Simultaneous analysis of alkamides and caffeic acid derivatives for the identification of Echinacea purpurea, Echinacea angustifolia, Echinacea pallida and Parthenium integrifolium roots. AB - A reversed-phase HPLC method was developed using a computer simulation program for the identification of dried roots of Echinacea purpurea, E. angustifolia, E. pallida and Parthenium integrifolium. Hydrophilic and lipophilic compounds were analysed simultaneously leading to a two-fold decrease in analysis time compared to traditional HPLC methods. PMID- 12094313 TI - Studies on the molecular markers of rhizomes of some Alpinia species. AB - The rhizomes of Alpinia jianganfeng are used as a traditional Chinese medicine, Jian Gan Feng, to cure rheumatism in Guangdong, China. The rhizomes of some other species of the genus Alpinia such as A. japonica, A. suishaensis and A. nanchuanensis are also used as Jian Gan Feng in Southwest China. However, the identification of the original plants of the crude drugs is difficult. The internal transcribed spacers and the 5.8S coding region of nuclear ribosomal DNA of the four species were sequenced and analyzed. The DNA markers have been determined and they can be used for the molecular identification of these medicinal plants. PMID- 12094314 TI - [Surgical approaches for limbal stem cell deficiency]. AB - BACKGROUND: Stem cells are the ultimate source of the rapidly self-renewing corneal epithelium and are located in the basal layer of the limbal epithelium. A variety of diseases can compromise the stem cell pool, causing an entity called limbal stem cell deficiency. Several therapeutic strategies should be taken into account. METHODS AND RESULTS: Ocular surface defence should be restored prior to any kind of stem cell transplantation to assure a sufficient and stable tearfilm. Surgical strategies comprise superficial keratectomy with or without amniotic membrane transplantation, conjunctival-limbal autograft and keratolimbal allograft transplantation. A damaged limbal and corneal stroma can be reconstructed by amniotic membrane as a biological substrate. First encouraging results have been reported using tissue bio-engineered limbal epithelium on amniotic membrane for corneal surface reconstruction. CONCLUSION: The correct diagnosis and a restored ocular surface defence are a prerequisite for subsequent transplantation of limbal stem cells. Optimal patient selection for the respective surgical approach and systemic immunosuppression protocols to improve allograft survival need to be further investigated. PMID- 12094315 TI - [Congenital ptosis: amblyogenic refractive errors, amblyopia, manifest strabismus and stereopsis related to the types of ptosis. Data on 77 patients and review of the literature]. AB - BACKGROUND: Former reports on amblyogenic refractive errors, amblyopia and binocular vision in congenital ptosis usually comprise all forms of ptosis without any differentiation. This study is an analysis of different kinds of ptosis. PATIENTS AND METHODS: 154 eyes (98 ptotic eyes) of 77 patients with congenital ptosis aged > or = 1 year (56 unilateral ptoses: 45 simple, 1 with rectus superior paresis, 7 with Marcus Gunn's syndrome, 2 congenital oculomotor palsies, 1 unilateral fibrosis syndrome; 21 bilateral ptoses: 10 simple, 2 with bilateral double elevator paresis, 7 blepharophimosis syndromes and 2 bilateral fibrosis syndromes) were investigated concerning visual acuity, refractive error (94 % in cycloplegy), strabismus and stereo acuity. As amblyogenic refractive errors were defined: astigmatism > or = ldpt, anisometropia > or = 1 dpt (spherical equivalent) and hyperopia > or = 3 dpt; amblyopia was defined as visual acuity less than 1.0 or a difference between both eyes of at least 0.2. RESULTS: Altogether were found: Hyperopia > or = 3 dpt in 28.6 % (n = 28); astigmatism > or = 1 dpt in 63.3 % (n = 62), anisometropia in 27.3 % (n = 21), amblyopia in 65.3 % (n = 64), strabismus in 29.9 % (n = 23) and stereopsis in 76.6 % (n = 59). In unilateral simple ptosis, astigmatism of the fellow eye was found in 26.8 % (n = 14). In unilateral ptosis, esotropia was 21.4 % (n = 12), in bilateral ptosis, exotropia 19 % (n = 4), as well as astigmatism > or = 3 dpt 26.2 % (n = 11). As was to be expected, ptosis groups with motility disorders were associated with higher rates of strabismus and amblyopia. The blepharophimosis syndrome could be differentiated by the typical lid anomalies. CONCLUSION: The subgroups of congenital ptosis differ in frequency of amblyogenic factors. In unilateral ptosis these are also frequent in the fellow eye. PMID- 12094316 TI - [Primary diagnosis of syphilis by the ophthalmologist]. AB - BACKGROUND: Throughout the developed world the reported incidence of sexually transmitted diseases is rising with previously unreported cases at rates of 40 - 70 %. According to German sources, recent epidemiological data show a considerable increase of syphilis especially among homosexual men in larger cities. PATIENTS: We report on 4 patients (age 38 - 54) who were referred to our outpatient department because of vision loss of unknown origin. All were subsequently tested positive for syphilis. None of the patients (3 men, 1 woman) belonged to a risk group, only one described systemic symptoms (urethritis and arthritis). The ocular manifestations of syphilis were broad: granulomatous and non-granulomatous anterior uveitis, papillitis, and chorioretinitis. Two patients also tested positive for HIV. After systemic antibiotic therapy, the ocular diseases stabilized. CONCLUSION: Diagnosis and therapy of syphilis is cost effective. The rise of syphilis especially in urban areas necessitates a high level of suspicion dealing with patients with intraocular inflammation of unknown origin. Lues serology should be incorporated into routine lab diagnostics to aid in the detection of such cases. PMID- 12094317 TI - [Trabecular aspiration in pseudoexfoliative glaucoma--surgery to primarily reduce intraocular pressure]. AB - BACKGROUND: Trabecular aspiration has been discussed during the past few years as a new surgical method in the treatment of pseudoexfoliative glaucoma. In this procedure PEX-material, pigment and detritus are aspirated from the trabecular meshwork. Trabecular aspiration has been evaluated mainly in combination with cataract extraction. This study reports on our first experiences concerning the IOP-reducing effect of trabecular aspiration as a primary surgical method in the management of pseudoexfoliative glaucoma. MATERIALS: 17 eyes of 14 patients (7 m, 7 f; 12 OD, 5 OS; age 71 +/- 6 years) with pseudoexfoliative glaucoma were included in this study and operated on by standardised trabecular aspiration (vacuum max. 200 mm Hg, 180 - 270 degrees, 5 min). Therapy success was defined as an IOP reduction by more than 20 % and less than 21 mm Hg. RESULTS: Therapy success was 82 % (14 out of 17) on the first postoperative day, 50 % after 30 days (8 out of 16) and 23 % after 180 days (3 out of 13). IOP was 26.8 +/- 8.5 mm Hg before surgical intervention, 18.1 +/- 11.4 mm Hg after 1 day, 19.1 +/- 7.9 mm Hg after 30 days and 19.2 +/- 5.2 mm Hg after 180 days. Mean quantity of antiglaucomatous eye drops application was 3.1 +/- 0.9 preoperatively, 0.9 +/- 1.6 after 1 day, 0.8 +/- 1.2 after 30 days and 1.0 +/- 1.3 after 180 days. CONCLUSIONS: Trabecular aspiration achieves a good short-term effect in the reduction of IOP in patients with pseudoexfoliative glaucoma. However, this effect was limited to a few weeks in most patients. Trabecular aspiration as a primary surgical method in the management of pseudoexfoliative glaucoma does not appear to be suitable for long-term IOP reduction. PMID- 12094318 TI - [Highly refractive silicone lens with sharp optic edge (CeeOn Edge, Model 911): one year results of a multicenter clinical trial on performance and efficacy]. AB - PURPOSE: To investigate the performance and efficacy of a new high refractive index (1.46) silicone intraocular lens with sharp optic edges. MATERIALS AND METHODS: In this open prospective multicenter study 149 patients underwent cataract surgery and implantation of CeeOn Edge in one eye. Best corrected visual acuity was determined at day 1 - 2, week 1 - 2, month 6 and month 12 postoperatively. Posterior capsule was clinically evaluated according to existence or absence of posterior capsule opacification (PCO). PCO was defined as relevant if the visual acuity loss exceeded at least two lines. The intraoperative handling of the lens was assessed concerning foldability, haptic handling, positioning, unfolding and general assessment. RESULTS: Visual acuity increased from preoperatively 0.3 to 0.9 after one year. PCO with impact on visual acuity (loss of 2 lines or more) was recorded in only two patients, after 2 weeks (YAG) resp. 1 year (no YAG). The handling and implantation was considered to be good to very good. No lens-related complications were seen during the one year study period. CONCLUSIONS: CeeOn Edge, Model 911, restores visual acuity after cataract surgery and shows promising handling results. The low rate of PCO and YAG is most probably related to the sharp edge of the lens. PMID- 12094319 TI - [Laser epithelial keratomileusis (LASEK): histological investigation for vitality of corneal epithelial cells after alcohol exposure]. AB - BACKGROUND: Laser epithelial keratomileusis (LASEK) is a new surgical procedure to treat myopia. An epithelial flap is created after the exposure to 20 % alcohol and following the laser ablation the epithelium is repositioned to its original location. The advantage of LASEK is that the ablated corneal surface is covered by a full thickness epithelium immediately after surgery. It is hypothesised that this epithelial coverage inhibits the wound healing response of the cornea. However, this concept will only work if the epithelial cells are still vital after the exposure to alcohol. MATERIAL AND METHODS: The vitality of the corneal epithelial cells was investigated in 6 human cadaver eyes after the exposure to 20 % alcohol over 15 to 60 s. The vitality of the corneal epithelial cells was assessed by soaking the specimen in a 0.1 % trypan blue solution and incubated at 37 degrees C for 2 min. After a wash with BSS the specimen were re-incubated at 37 degrees C for 30 min in culture medium. After one more wash with BSS the cells were observed with a standard light microscope. Cells which retained the blue colour would be dead and vital cells would appear clear. Light microscopy and transmission electron microscopy was used to detect morphological changes. RESULTS: Mainly vital corneal epithelial cells were seen for an alcohol exposure time of up to 30 s. In particular the basal epithelial cells appeared alive. With a exposition time of 60 s most cells were dead. CONCLUSION: In LASEK the exposure time of 20 % alcohol is between 20 and 30 s. Based on our results we can conclude that after such exposure time most cells are alive, which is essential for the postoperative wound healing response. PMID- 12094320 TI - [Foveal cone dystrophy: diagnostic ranking of the multifocal electroretinogram]. AB - BACKGROUND: In case of visual loss without morphologic pathology differentiation between a functional visual loss and a retinal disorder may be difficult. PATIENTS AND METHODS: We report on two women, aged 33 and 44, who were complaining of a progressive visual loss occurring over the past several weeks to months. They were referred to our department with the presumed diagnosis of functional visual loss. Biomicroscopy of the anterior and posterior segments, perimetry (Goldmann), colour (FM-100-Hue-Test) and stereopsis testing (Lang Stereotest), fluorescein angiography (FLAG) and electrophysiological testing (P ERG, VECP, EOG, multifocal electroretinography [mfERG]) were performed. RESULTS: Visual acuity was 0.16 OU and 0.25 OU respectively. A small relative central scotoma (I/1) was detected by dynamic perimetry. Colour vision, random-dot stereopsis and the results of electrophysiological testing except multifocal electroretinography (mfERG) were normal. The mfERG was able to detect a cone dysfunction which was related to the fovea. CONCLUSION: In case of visual loss of unknown etiology electrophysiological testing is indicated. The mfERG as an objective diagnostic method is able to detect a foveal cone dystrophy. PMID- 12094321 TI - [Pyogenic granuloma of the lacrimal sac]. AB - BACKGROUND: Pyogenic granulomata belong to the group of inflammatory pseudotumours. They consist of granulation tissue usually following inflammatory processes or trauma. PATIENT: We report on a 86-year old female suffering from chronic dacryocystitis. In the course of the treatment exstirpation of the lacrimal sac was performed. The removed tissue was analysed histologically. RESULTS: Histological examination revealed a dense lymphoplasmacellular infiltration underlying the epithelium of the lacrimal sac. A mushroom-like tumour rising into the lumen of the lacrimal sac could be observed. This tumour consisted of capillaries and a dense inflammatory infiltrate. The findings were consistent with a pyogenic granuloma of the lacrimal sac. CONCLUSION: Pyogenic granulomata usually arise in the conjunctiva as a sequel of chronic lipogranulomatous inflammation. They are very rare in other areas such as the lacrimal drainage system. PMID- 12094322 TI - [Angiolymphoid hyperplasia with eosinophilia with bilateral involvement of the lacrimal glands]. AB - BACKGROUND: Angiolymphoid hyperplasia with eosinophilia (ALHE) and Kimura's disease are two rarely occurring disorders very similar to each other; however, they are individual nosological entities. For a long time they were considered as a single disease due to the frequency of similar characteristics. The majority of authors have recently considered these diseases as two individual entities with some reciprocal specifications, both clinical as well as morphological. PATIENT: We report on the case of a 76-year old male white European who had suffered for more than six years from subcutaneous tumour formations in different parts of the body. Eyelid oedema on the right side with palpable resistance under the upper temporal edge of the orbit initially occurred five years after the first symptoms of the disease. A similar finding occurred on the left side after eight months. The tumours on both sides were surgically removed and sent for histological analysis. A unilateral recurrence of the finding appeared after one year, followed by surgical intervention and histological examination. RESULTS: Tumour infiltrate in the first two orbita excisions was topically related to lacrimal gland structures; these structures were not found in the third excision. The case was histologically diagnosed as ALHE despite some similar characteristics with Kimura's disease. The diagnosis of ALHE was histologically supported, especially by the absence of lymphatic follicle formations and fibrotisation in the infiltrate, and the identification of the appearance of epitheloid up to histiocytoid of proliferating endothelium, which forms small lumens. The diagnosis of ALHE was additionally supported by the fact that the incidence of Kimura's disease in white Europeans is very rare. CONCLUSIONS: According to our findings, both entities overlap one another, even in some characteristics considered to be distinguishing. The question arises, therefore, whether the strict separation of these nosological entities can indeed be determined with one hundred percent accuracy. PMID- 12094323 TI - [Bilateral nevus of the caruncle]. AB - BACKGROUND: Pathologic processes of the caruncle are relatively rare with a predominance of papillomas, pyogenic granulomas and nevi. PATIENT: A 22 year old Caucasian male patient presented with bilateral hyperpigmentations of the caruncle. They had first been observed at an age of seven and showed slight progression in size and colour with time. Examination revealed hyperpigmentation with clear pseudocystic spaces. The tarsal or bulbar conjunctiva was not involved. An excision of both processes was performed. HISTOLOGY: Light microscopy revealed nests of nevus cells with melanin granules and round basophilic nuclei as well as gland ducts and sebaceous glands, beneath and within the multilayered keratinizing and nonkeratinizing squamous epithelium. The sections revealed a bilateral melanocytic compound nevus of the caruncle. DISCUSSION: To our knowledge, there is no report on bilateral nevi of the caruncle among the Caucasian population. The possibility of a common precursor cell during the embryologic development causing the bilaterality is discussed. PMID- 12094324 TI - [Successful topical cyclosporin A in the therapy of progressive vascularising keratitis in keratitis-ichthyosis-deafness (KID) syndrome (Senter syndrome)]. AB - BACKGROUND: The keratitis-ichthyosis-deafness (KID) syndrome (Senter Syndrome) and its major criteria erythrokeratodermia, neuro-sensoric deafness and vascularising keratitis were defined in 1981. Several cases have been described since 1915, but up to now no causal therapy of this disease with sporadic occurrence has been found. Clinical experience using systemic Cyclosporin A (CsA) in the dermatological therapy of the KID syndrome differs. Up to now there has been no report on the use of topical CsA eye drops in the therapy of vascularising keratitis. HISTORY AND SIGNS: We report on a case of keratitis, ichthyosis and deafness (KID syndrome) in a 36-year old male. Both eyes were suffering from recurrent corneal ulcers (ulcus serpens corneae). No improvement had been observed on topical or systemic medication (including lubricants, antibiotics, steroids, etc.) during a history of more than two years. THERAPY AND OUTCOME: Topical CsA 2 % (eye drops, 3 times daily) was administered in addition to lubricants. After 3 months the deep corneal neovascularisations were significantly reduced. The corneal ulcer was closed and visual acuity increased from 20/400 (OU) to 20/100 (OD) and 20/200 (OS), respectively. CONCLUSION: A dose dependent keratinocyte growth inhibition was found in vitro for cyclosporin A using ichthyosis skin samples. The reduction of activated T-lymphocytes and the resulting reduced HLA-Class-2 expression of conjunctiva and cornea as well as the inhibition of activated T-Cells in the lacrimal gland might be responsible. Nevertheless, the effect of topical CsA on the vascularizing keratitis in the KID syndrome is unknown. PMID- 12094325 TI - [Spontaneous regression of an eyelid tumour]. AB - BACKGROUND: The differential diagnosis of eyelid tumours is often a challenging task. A correct anamnesis can be very important for the further treatment plan. CASE REPORT: A 73-year-old female patient presented with a tumour of her left lower eyelid which occurred two months ago. The suspected clinical diagnosis of a basal cell carcinoma was confirmed. After one month the patient was admitted for tumour excision and eyelid reconstruction, but the tumour showed marked regression. Because of a now clinically suspected keratoacanthoma, we decided to observe the tumour and cancelled surgery. Fourteen days later there was no tumor visible on her left lower eyelid. It seemed to be a spontaneous regression of a keratoacanthoma. CONCLUSIONS: If there is a short case history of a rapidly growing eyelid tumour one should also consider keratoacanthoma as differential diagnosis. In suspected cases initial observation and control can save the patient from extended excision and eyelid reconstruction. However, an unnecessary delay of treatment due to prolonged observation without any signs of regression should be avoided. Unclear situations require a diagnostic excision to exclude a squamous cell carcinoma or basal cell carcinoma. PMID- 12094327 TI - Molecular characterization of the pericentric inversion that causes differences between chimpanzee chromosome 19 and human chromosome 17. AB - A comparison of the human genome with that of the chimpanzee is an attractive approach to attempts to understand the specificity of a certain phenotype's development. The two karyotypes differ by one chromosome fusion, nine pericentric inversions, and various additions of heterochromatin to chromosomal telomeres. Only the fusion, which gave rise to human chromosome 2, has been characterized at the sequence level. During the present study, we investigated the pericentric inversion by which chimpanzee chromosome 19 differs from human chromosome 17. Fluorescence in situ hybridization was used to identify breakpoint-spanning bacterial artificial chromosomes (BACs) and plasmid artificial chromosomes (PACs). By sequencing the junction fragments, we localized breakpoints in intergenic regions rich in repetitive elements. Our findings suggest that repeat mediated nonhomologous recombination has facilitated inversion formation. No addition or deletion of any sequence element was detected at the breakpoints or in the surrounding sequences. Next to the break, at a distance of 10.2-39.1 kb, the following genes were found: NGFR and NXPH3 (on human chromosome 17q21.3) and GUC2D and ALOX15B (on human chromosome 17p13). The inversion affects neither the genomic structure nor the gene-activity state with regard to replication timing of these genes. PMID- 12094328 TI - A CHEK2 genetic variant contributing to a substantial fraction of familial breast cancer. AB - CHEK2 (previously known as "CHK2") is a cell-cycle-checkpoint kinase that phosphorylates p53 and BRCA1 in response to DNA damage. A protein-truncating mutation, 1100delC in exon 10, which abolishes the kinase function of CHEK2, has been found in families with Li-Fraumeni syndrome (LFS) and in those with a cancer phenotype that is suggestive of LFS, including breast cancer. In the present study, we found that the frequency of 1100delC was 2.0% among an unselected population-based cohort of 1,035 patients with breast cancer. This was slightly, but not significantly (P=.182), higher than the 1.4% frequency found among 1,885 population control subjects. However, a significantly elevated frequency was found among those 358 patients with a positive family history (11/358 [3.1%]; odds ratio [OR] 2.27; 95% confidence interval [CI] 1.11-4.63; P=.021, compared with population controls). Furthermore, patients with bilateral breast cancer were sixfold more likely to be 1100delC carriers than were patients with unilateral cancer (95% CI 1.87-20.32; P=.007). Analysis of the 1100delC variant in an independent set of 507 patients with familial breast cancer with no BRCA1 and BRCA2 mutations confirmed a significantly elevated frequency of 1100delC (28/507 [5.5%]; OR 4.2; 95% CI 2.4-7.2; P=.0002), compared with controls, with a high frequency also seen in patients with only a single affected first-degree relative (18/291 [6.2%]). Finally, tissue microarray analysis indicated that breast tumors from patients with 1100delC mutations show reduced CHEK2 immunostaining. The results suggest that CHEK2 acts as a low-penetrance tumor suppressor gene in breast cancer and that it makes a significant contribution to familial clustering of breast cancer-including families with only two affected relatives, which are more common than families that include larger numbers of affected women. PMID- 12094329 TI - Evidence consistent with human L1 retrotransposition in maternal meiosis I. AB - We have used a unique polymorphic 3' transduction to show that a human L1, or LINE-1 (long interspersed nucleotide element-1), retrotransposition event most likely occurred in the maternal primary oocyte during meiosis I. We characterized a truncated L1 retrotransposon with a 3' transduction that was inserted, in a Dutch male patient, into the X-linked gene CYBB, thereby causing chronic granulomatous disease. We used the unique flanking sequence to localize the precursor L1 locus, LRE3, to chromosome 2q24.1. In a cell culture assay, the retrotransposition frequency of LRE3 is greater than that for any other element that has been tested to date. The patient's mother had two LRE3 alleles that differed slightly in the 3'-flanking genomic DNA. The patient had a single LRE3 allele that was identical to one of the maternal alleles; however, the patient's insertion matched the maternal LRE3 allele that he did not inherit. Other data indicate that there is only a small chance that the father (unavailable for analysis) carries the precursor LRE3 allele. In addition, paternal origin of the insertion would have required that an LRE3 mRNA transcribed before meiosis II be carried separately from its precursor LRE3 allele in the fertilizing sperm. Since the mother carries a potential precursor allele and the insertion was on the patient's maternal X chromosome, it is highly likely that the insertion originated during maternal meiosis I. PMID- 12094330 TI - A genetic locus for adolescent idiopathic scoliosis linked to chromosome 19p13.3. AB - Adolescent idiopathic scoliosis (AIS) is one of the most common orthopedic disorders, affecting up to 4% of schoolchildren worldwide. We studied seven unrelated multiplex families of southern Chinese descent with AIS, consisting of 25 affected members. A genomewide scan with >400 fluorescent microsatellite markers was performed. Multipoint linkage analysis by GENEHUNTER revealed significant linkage of the abnormal phenotype to the distal short arm of chromosome 19, with both a maximum multipoint LOD score and a nonparametric LOD score of 4.93. Two-point linkage analysis by MLINK gave a LOD score of 3.63 (recombination fraction theta[m=f]=0.00) at D19S216. Further high-density mapping and informative recombinations defined the AIS critical region in the vicinity of D19S216, flanked by D19S894 and D19S1034, spanning 5.2 cM on the sex-averaged genetic map on chromosome 19p13.3. PMID- 12094333 TI - Present and future treatment of advanced non-small cell lung cancer. AB - Platinum-based chemotherapy is considered the standard treatment for advanced non small cell lung cancer (NSCLC). Several phase II trials using cisplatin in combination with new chemotherapeutic agents, such as gemcitabine, the taxanes, vinorelbine, and irinotecan, showed impressive response rates and suggested an improvement in overall survival. Large phase III trials comparing these second generation cisplatin regimens indicated a substantial equivalence of new combinations, marginally improving the outcome of patients over the first generation platinum-based regimens. Phase III trials have not yet shown dramatic advantages for either multiple-drug regimens, with nonoverlapping mechanisms of action and toxicity, or nonplatinum doublets, with efficacy and/or toxicity profiles superior to those of platinum-based chemotherapy. Chemotherapy in advanced non-small cell lung cancer has reached a plateau, and it is clear that new approaches are required. These should include prevention, screening, and early detection, and the use of novel treatments based on our understanding of the biology and molecular biology of this disease. PMID- 12094332 TI - Expression of target molecules in lung cancer: challenge for a new treatment paradigm. AB - Lung cancer is the leading cause of cancer death in men and women in the United States, accounting for 28% of all cancer fatalities. More than two thirds of patients present with metastatic disease at the time of presentation. Despite improvements in chemotherapy and combined treatment modalities, the survival rate remains below 15%. However, recent advances in our understanding of the biology of lung cancer and carcinogenesis have led to the development of novel therapies directed at tumor-specific targets. These targets are crucial components in important pathways for cell growth, proliferation, and apoptosis. Strategies that interfere with these pathways include monoclonal antibodies directed at growth factors or their receptors, immunotoxins, ligand toxins, antisense molecules, ribozymes, and small-molecule inhibitors. Novel cell surface antigens are being used in vaccines developed to stimulate T-cell-specific immunity. The tumor cells also have specific survival requirements in their local environment that are necessary for invasion, angiogenesis, and metastases. Many new therapeutic strategies are designed to interfere with these requirements. This article reviews many of these recent developments and new therapeutic possibilities; ideally, in the near future, these developments will be implemented in the treatment of lung cancer patients and in early detection and chemoprevention strategies. PMID- 12094334 TI - Incorporation of pemetrexed (Alimta) into the treatment of non-small cell lung cancer (thoracic tumors). AB - Lung cancer is the leading cause of cancer death in the United States and throughout the world. The overall 5-year survival rate for lung cancer is dismal: 14% in the United States and even lower in other parts of the world. Recent developments in the armamentarium of chemotherapeutic agents for lung cancer have shown that two-drug combinations improve survival, relieve symptoms, and improve quality of life; however, complete response rates are still approximately 1% in stage IV disease and less than 20% of advanced stage patients survive 2 years. Therefore, improved therapeutic agents that increase efficacy are sorely needed. Most lung cancers overexpress thymidylate synthase and a variety of genes involved in cell cycle regulation. Previous studies have shown that some inhibitors of DNA synthesis (eg, gemcitabine) can improve the survival of advanced lung cancer patients, especially when combined with other agents such as cisplatin. The multitargeted antifolate, pemetrexed (Alimta; Eli Lilly and Co, Indianapolis, IN) was developed because it inhibits multiple enzymes involved in DNA synthesis including thymidylate synthase, dihydrofolate reductase, and glycinamide ribonucleotide formyl transferase. The early studies of pemetrexed showed that the important dose-limiting toxicities were myelosuppression, mucositis, and diarrhea, all of which are common with any antimetabolite. Subsequent studies described in this article will show that these toxicities can be significantly reduced by the use of vitamin supplementation with folate and B12, and that pemetrexed has considerable activity in non-small cell lung cancer and mesothelioma. PMID- 12094335 TI - Present and future treatment of pancreatic cancer. AB - Gemcitabine has become a new standard for treatment of advanced pancreatic cancer. This development is based not only on drug efficacy but also on a favorable side-effect profile. Combinations of gemcitabine with antitumor drugs such as cisplatin, 5-fluorouracil, docetaxel, irinotecan, oxaliplatin, or capecitabine, and biological agents such as cetuximab or trastuzumab, have yielded promising results in phase II trials. However, none of these combinations has yet reached the level of an evidence-based standard treatment. PMID- 12094336 TI - The present and future of combination chemotherapy in bladder cancer. AB - The combination of methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC) has dominated the landscape of chemotherapy for advanced bladder cancer for over 15 years. Randomized studies have shown its superiority over cisplatin alone or in combination with cyclophosphamide and doxorubicin. However, it exhibits a significant toxicity profile and achieves only a slight impact on overall survival. Gemcitabine is among the new cytotoxic drugs in development for treatment of advanced urothelial cancer. The combination of gemcitabine and cisplatin represents a new standard alternative of treatment in the disease based on similar efficacy to and lower toxicity than the classic MVAC regimen. Future drug development will focus on the clinical usefulness of three-drug regimens (including gemcitabine, paclitaxel or docetaxel, and a platinum salt), and nonplatinum-based combinations, as well as showing the impact of adjuvant postcystectomy chemotherapy on survival. PMID- 12094337 TI - Pemetrexed: translational research in breast cancer. AB - Pemetrexed (Alimta; Eli Lilly and Co, Indianapolis, IN) is a novel antifolate/antimetabolite with activity in breast cancer and has well-defined molecular targets, including thymidylate synthase, dihydrofolate reductase, and glycinamide ribonucleotide formyltransferase. In a phase II trial in patients with T3-4, N0-2 breast cancer, expression of thymidylate synthase, dihydrofolate reductase, glycinamide ribonucleotide formyltransferase, p53, and c-erb-B2 (at the mRNA or protein level) is being examined before and 24 hours after the first dose of neoadjuvant pemetrexed and after three cycles of single-agent treatment to establish correlations of biomarker levels and changes with clinical outcome and toxicity. Full biomarker and clinical data are not yet available from this study; however, clinical responses to pemetrexed treatment have been observed in patients. Results of this trial should provide both an idea of the activity of neoadjuvant pemetrexed in breast cancer and information on biomarker association with clinical performance that can be used in the design of additional clinical studies to assess the predictive value of these markers. PMID- 12094338 TI - Gemcitabine and pemetrexed (Alimta) in thoracic cancers: present and future perspectives. Introduction and current controversies. AB - With more than 1 million new cases annually worldwide, lung cancer remains a major public health problem, and ongoing efforts against tobacco consumption will not modify this situation in the near future. Most patients with lung tumors have advanced disease at the time of presentation and are thus treated with palliative intent. Several new active cytotoxic agents (ie, gemcitabine, irinotecan, the taxanes, and vinorelbine) have emerged during the last decade, and in combination with cisplatin, offer response rates of 35% to 50% and median survival durations of 9 to 11 months. Nevertheless, the results remain disappointing in the majority of cases, and data on long-term survivors are still anecdotal. Several strategies might improve treatment efficacy: administering multidrug combinations to increase antitumor activity, ameliorating the tolerance of chemotherapy, treating patients at an earlier stage of their disease, and developing new agents. This article discusses these promising strategies and places them in proper perspective. PMID- 12094339 TI - Early results of a randomized phase III trial of platinum-containing doublets versus a nonplatinum doublet in the treatment of advanced non-small cell lung cancer: European Organization for Research and Treatment of Cancer 08975. AB - In the European Organization for Research and Treatment of Cancer randomized phase III trial (08975), 480 patients with advanced non-small cell lung cancer received the standard arm of cisplatin/paclitaxel or one of two experimental arms; gemcitabine/cisplatin or gemcitabine/paclitaxel. Cisplatin was given at 80 mg/m(2) on day 1, gemcitabine was administered at 1,250 mg/m(2) on days 1 and 8, and paclitaxel at 175 mg/m(2) as a 3-hour infusion on day 1, every 3 weeks. The cisplatin/paclitaxel and gemcitabine/cisplatin regimens were comparably active. The nonplatinum arm, gemcitabine/paclitaxel, was well tolerated, but had a trend toward lower median, 1-year, and progression-free survival compared with the standard arm. Further follow-up and analysis of quality of life may clarify this possible difference in treatment outcome. PMID- 12094340 TI - Gemcitabine versus gemcitabine/carboplatin in advanced non-small cell lung cancer: preliminary findings in a phase III trial of the Swedish Lung Cancer Study Group. AB - Gemcitabine is an active agent in non-small cell lung cancer, with single-agent treatment producing response rates of approximately 20% and median survivals of approximately 7 to 9 months. In a pilot trial in advanced non-small cell lung cancer, the gemcitabine/carboplatin combination produced a response rate of 43% and median survival of 12 months with good tolerability. Preliminary results of a phase III trial comparing gemcitabine alone with gemcitabine/carboplatin in 332 patients with stage IIIB or IV non-small cell lung cancer are now available. Patients were randomized to receive gemcitabine 1,250 mg/m(2) on days 1 and 8 every 21 days or the same gemcitabine regimen plus carboplatin at an area under the concentration-time curve of 5 mg/mL/min on day 1 for a maximum of six cycles. Hematologic toxicity was more common in the combination arm; grade 4 thrombocytopenia occurred in 23.5% v 5.3% of patients, but infrequently resulted in clinical complications. Nonhematologic toxicity was moderate and similar in frequency in the combination and gemcitabine arms (25% and 28%, respectively). Among 275 patients, overall response rates were 30% (2% complete response and 28% partial response) in the combination arm and 12% (all partial responses) in the gemcitabine arm. Median time to disease progression was 6 months in the combination arm and 4 months in the gemcitabine arm. Median survival in the study population was 9 months, a promising finding given the high proportion of elderly patients in the study (37% >/= 70 years of age). Full mature results of the trial, including comparative survival results and data on quality of life, are awaited. PMID- 12094341 TI - Gemcitabine induction chemotherapy in non-small cell lung cancer. AB - Surgery has long been considered standard treatment in early stage non-small cell lung cancer. Preoperative chemotherapy is a real challenge in the treatment of these stages. Some conclusions can be drawn from the first phase II studies in stage IIIA tumors. Response rates were higher than those observed in stage IV tumors, reaching approximately 70%. Although toxicity seemed acceptable, increased morbidity and mortality have to be taken into account for the choice of preoperative regimens. Two randomized studies that included only a few patients were conducted in stage IIIA disease and showed highly positive survival results. The French randomized study argued in favor of preoperative chemotherapy with an absolute difference in survival rates that remains constant beyond the third year. New studies are ongoing to evaluate the role of the gemcitabine/cisplatin combination. Several phase II studies of this regimen in the preoperative setting or in combination with radiotherapy have been presented at the most recent meetings of the American Society of Clinical Oncology. These studies confirmed both its efficacy and good tolerability. In several ongoing randomized studies, this combination has been chosen to test the concept of preoperative chemotherapy. One such study, which compares two different strategies of preoperative chemotherapy, is by the Intergroupe Francophone de Cancerologie Thoracique. PMID- 12094342 TI - Nasal dermoid sinus cysts in the dog. AB - OBJECTIVE: To describe the clinical and morphologic characteristics of nasal dermoid sinus cysts in the dog. STUDY DESIGN: Retrospective clinical study. ANIMALS: Six client-owned dogs. METHODS: Medical records (1995 to 1999) of 6 dogs that had a discharging sinus in the midline of the nose removed surgically were reviewed for signalment, physical examination, surgical and histopathologic findings, and outcome. RESULTS: Three golden retrievers, 1 springer spaniel, 1 cocker spaniel, and 1 Brittany spaniel with a history of intermittent discharge from a small opening in the midline on the bridge of the nose at the junction between the nasal planum and the skin were identified. The condition was unresponsive to antibiotic therapy, and previous surgical excision had been unsuccessful in 3 dogs. None of the dogs had any other congenital defects, and all dogs responded to complete surgical excision of the tract and cyst. On histopathologic examination of excised tissue, there were adnexal structures along a tract lined with stratified squamous epithelium, consistent with a developmental abnormality of ectodermal tissue. CONCLUSIONS: The lesions were diagnosed as nasal dermoid sinus cysts, similar to the congenital condition described in humans. Nasal dermoid sinus cysts are rare developmental defects related to abnormal development of the pre-nasal space and may extend into the cranial vault causing cerebral abscesses or recurrent meningitis. Complete surgical excision has a good prognosis. CLINICAL RELEVANCE: This is a new condition that should be added to the surgical differential diagnosis for a discharging sinus over the external nares in dogs. PMID- 12094343 TI - Effects of 25% propylene glycol hydrogel (Solugel) on second intention wound healing in horses. AB - OBJECTIVE: To evaluate the effect of a commercially available 25% propylene glycol hydrogel preparation (Solugel; Johnson and Johnson Medical, North Ryde, Australia) on healing of full-thickness skin wounds on the distal aspect of the limb in horses. STUDY DESIGN: Experimental. ANIMALS: Eight Standardbred horses. METHODS: Standardized (2.5 x 2.5 cm) full-thickness skin wounds were created over the mid-dorsomedial aspect of both metacarpi in 8 horses. One wound in each horse was dressed with saline solution (0.9% NaCl) soaked gauze, and one was treated with Solugel under dry regular gauze; wounds were then bandaged with gauze-coated cotton wool and elastic adhesive bandages. Wounds were videorecorded and rebandaged twice weekly until healed. Wound healing variables were measured from the videorecordings using a computer software package and analyzed as a randomized complete block design with repeated measures. Where necessary variables were made positive for analysis; significance was set at P <.05. RESULTS: The area of the wound at the first bandage change did not vary between treated and untreated wounds. Treatment had no effect on the total rate of healing, rate of healing during the retraction phase of healing, rate of healing after the retraction phase was complete, or the amount the wounds retracted. CONCLUSIONS: Using this model of wound healing, Solugel had no effect on second intention healing of distal limb wounds in horses. CLINICAL RELEVANCE: Solugel does not appear to have any beneficial effect on healing of small full-thickness skin wounds to the distal limb of horses. PMID- 12094344 TI - The effect of equine recombinant growth hormone on second intention wound healing in horses. AB - OBJECTIVE: To evaluate the effect of intramuscular administration of recombinant equine growth hormone on healing of full thickness skin wounds on equine limbs. STUDY DESIGN: Experimental. ANIMALS: Nine Standardbred horses. METHODS: In study 1, standardized full thickness skin wounds (2.5 x 2.5 cm) were made over the dorsomedial aspect of the mid-cannon bone of 1 forelimb and 1 hindlimb in 9 horses. Wounds were bandaged without treatment (control subjects) and videorecorded twice weekly until healed. Then, in study 2, similar wounds were created on the opposite limbs; 6 horses were administered intramuscular recombinant equine growth hormone (10 microg/kg daily for 7 days, then 20 microg/kg daily for 49 days), and 3 horses (control subjects) were administered equivalent volumes of sterile water. Wounds were videorecorded twice weekly until healed. Wound healing variables were measured from the videorecordings using a computer software package and analyzed as a randomized complete block design factorial analysis of variance; significance was set at P <.05. RESULTS: No differences in the measured variables were detected between wounds in study 1 and the control wounds in study 2. In recombinant equine growth hormone-treated horses, wounds retracted more during treatment and contracted faster after treatment stopped when compared with wounds from untreated horses. No other treatment effects were detected. CONCLUSIONS: Recombinant equine growth hormone seemingly increases wound retraction. After treatment ceases, wound contraction increases. CLINICAL RELEVANCE: Intramuscular administration of recombinant equine growth hormone (10 microg/kg daily for 7 days, then 20 microg/kg daily for 49 days) does not appear to have any beneficial clinical effect on healing of equine limb wounds. PMID- 12094345 TI - The effect of recombinant equine growth hormone on the biomechanical properties of healing superficial digital flexor tendons in horses. AB - OBJECTIVE: To evaluate the effect of recombinant equine growth hormone (rEGH) on the in vitro biomechanical properties of healing superficial digital flexor tendon (SDFT) in horses. STUDY DESIGN: Completely randomized design. SAMPLE POPULATION: Twelve Standardbred horses, 3 to 7 years of age, with ultrasonographically normal forelimb SDFT. METHODS: One week after induction of collagenase (2,000 U) induced superficial flexor tendonitis, horses were randomly divided into groups of 6. One group was administered intramuscular rEGH (10 microg/kg/day for 1 week, then 20 microg/kg/day for 5 weeks), whereas the other group (control subjects) were administered an equivalent volume of saline (0.9% NaCl) solution. At the end of this 6-week treatment, horses were killed and one forelimb SDFT from each horse was harvested for biomechanical testing under uniaxial tension. Results were analyzed using an unpaired Student's t test; significance was set at P 7 mg/dL was killed at 51 days postoperatively. In the cyclosporine-alone group: 3 dogs with acute rejection were killed at 6 to 9 days and 3 dogs survived to 100 days. In dogs treated with cyclosporine and MNA 715, survival to histologically confirmed acute rejection was significantly longer (P =.044) and the degree of mononuclear cell infiltration was significantly reduced (P =.040), compared with dogs treated with cyclosporine alone. CONCLUSIONS: MNA 715 combined with cyclosporine prolonged allograft survival and reduced the severity of histologic rejection in a clinically relevant renal transplant model. CLINICAL RELEVANCE: An immunosuppressive regimen consisting of MNA715 and microemulsified cyclosporine may be effective in preventing allograft rejection in canine renal transplant patients. PMID- 12094351 TI - Are circular external fixators weakened by the use of hemispheric washers? AB - OBJECTIVES: To compare the axial mechanical stability of 3 circular external fixators systems with and without hemispheric washers. STUDY DESIGN: Experimental study. METHODS: The axial stiffness and load necessary to produce 0.5 and 1 mm of displacement of 10 circular external fixator constructs from 3 manufacturers were tested on a materials testing machine. The constructs tested included the Small Bone fixator (SBF; Hofmann S.a.S., Monza, Italy), the IMEX ring fixator (IMEX Inc., Longview, TX), and the Multiplanar C-Fix (MCF; PanVet Distribuzione, Seriate, Italy). Five configurations were tested for each construct: (1) conventional nut fixation, (2) hemispheric washer fixation with connecting rods offset by 0, (3) 1, and (4) 2 holes, and (5) with a ring placed at maximum angulation. RESULTS: The loads resisted at 0.5 and 1 mm of displacement did not differ when frame configurations were compared (P =.25733 and.33769, respectively). The linear stiffness of the following configurations were decreasingly stiff: standard constructs, hemispheric washers with connecting rods perpendicular to rings, hemispheric washers with connecting rods offset by 1 hole, hemispheric washers with connecting rods offset by 2 holes, and ring offset in relation to bone model. The SBF constructs tested were 34% and 41% more rigid than the IMEX and MCF constructs tested despite the larger diameter of the connecting rods for the IMEX frames (6 mm) compared with the SBF frames (5 mm). The IMEX constructs tested were 6% more rigid than the MCF constructs tested. CONCLUSIONS: Adding hemispheric washers and angling connecting rods in relation to rings did not influence the loads resisted at 0.5 and 1 mm displacement but decreased construct stiffness. CLINICAL RELEVANCE: The use of hemispheric washers had minor effects on the biomechanical performance of fixator frames tested in this study when used to angle a ring in relation to connecting rods for circular external fixators. PMID- 12094352 TI - Transendoscopic electrosurgery of an equine pulmonary granular cell tumor. AB - OBJECTIVE: Description of transendoscopic electrosurgical treatment of a pulmonary granular cell tumor in a horse. STUDY DESIGN: Case report. ANIMALS: A 12-year-old Standardbred mare. METHODS: After endoscopic examination and transendoscopic biopsy, a granular cell tumor nodule was removed transendoscopically from the right principal bronchus of the sedated horse (0.02 mg/kg detomide intravenous) with a monopolar electrosurgical wire snare. Two days later, the surgical area was irradiated transendoscopically using a Nd-YAG laser. RESULTS: After surgery, the horse had no further signs of respiratory disease. Clinical and endoscopic examination and radiography performed 4 times during the following 2 years showed no evidence of recurrence. CONCLUSIONS: Transendoscopic electrosurgical removal of nodular masses located intraluminally in the principal bronchi is possible on the sedated standing horse. CLINICAL RELEVANCE: Granular tumors are rare lung neoplasms that lead to unspecific signs of a low airway disease. Diagnosis requires endoscopic examination of the large bronchi. Early intraluminal stages can be treated transendoscopically. PMID- 12094353 TI - The use of porcine small intestinal submucosa as a biomaterial for perineal herniorrhaphy in the dog. AB - OBJECTIVES: To develop an in vivo perineal hernia model, to develop a technique for using small intestinal submucosa (SIS) in perineal hernia repair, to further elucidate the biological behavior of SIS, and to compare SIS herniorrhaphy with the internal obturator muscle transposition (IOT) technique. STUDY DESIGN: Prospective evaluation comparing SIS herniorrhaphy with IOT. ANIMALS: Twelve adult castrated male, large-breed dogs. METHODS: All dogs had bilateral pelvic diaphragm defects created by complete excision of the levator ani muscle. Each dog had one side repaired using SIS and the other by IOT. Pain and inflammation were subjectively scored. Dogs were killed 2 weeks (n = 4), 12 weeks (n = 4), or 16 weeks (n = 4) after surgery. Each pelvic diaphragm was biomechanically tested to failure. The pelvic diaphragms from 2 normal dogs (n = 4 sides) were also biomechanically tested. Failure site, maximum pressure, displacement at failure, and initial linear stiffness values were determined. Histologic assessment was performed. Statistical analysis was performed with significance set at P <.05 RESULTS: No significant postoperative complications were noted. There were no significant differences in maximum pressure to failure, displacement, or stiffness when comparing normal, SIS, and IOT at any time point. The SIS group had significantly less displacement (P =.004) at 2 weeks than at weeks 12 or 16. For all herniorrhaphy techniques, the failure site was central (n = 22) or at the suture line (n = 2). At 2 weeks, histologic evaluation of tissues from the IOT group showed inflammation, mineralization, and necrosis, which were not present in tissues from the SIS group. Histologic examination at 12 and 16 weeks showed no microscopic differences in cell population or tissue characteristics between the IOT and SIS groups. CONCLUSIONS: SIS herniorrhaphy was successfully performed in this in vivo model of perineal hernia in the dog. CLINICAL RELEVANCE: This study suggests that SIS can be used as a primary means of repair, as augmentation when the internal obturator muscle is thin and friable, or as a salvage procedure in cases of recurrence in dogs with perineal hernia. PMID- 12094354 TI - The effect of nephrotomy on renal function and morphology in dogs. AB - OBJECTIVE: To determine whether bisection or intersegmental nephrotomy adversely affects glomerular filtration rate (GFR) and whether bisection nephrotomy has less adverse effect than intersegmental nephrotomy. STUDY DESIGN: Dogs assigned to 5 groups of 3 dogs each, depending on time of euthanasia at postoperative day 1, 4, 8, 15, or 29. ANIMALS: Fifteen healthy adult intact female dogs. METHODS: Within 4 days of total and single kidney GFR measurements using technetium Tc 99m DTPA, left intersegmental nephrotomy and right bisection nephrotomy were performed. The GFR measurements were repeated the day before dogs were killed except for the day 1 group. Histologic examination was performed on kidneys harvested from day 1, 4, 8, and 29 groups. Severity of lesions was scored on a 0 to 4 scale. Single kidney GFR was compared in each kidney before and after nephrotomy. Histologic and GFR changes were compared between bisection and intersegmental nephrotomy kidneys for each time period. RESULTS AND CONCLUSIONS: Three days after surgery, total GFR was increased from baseline by 176%, with no difference in percent change in single kidney GFR between intersegmental and bisection nephrotomy. By day 7, total and single kidney GFR were not different than baseline. Bisection nephrotomy caused more intrarenal hemorrhage and cortical infarction and inflammation than intersegmental nephrotomy. By 4 weeks, no significant histologic differences were observed between the 2 techniques. CLINICAL RELEVANCE: Neither intersegmental nor bisection nephrotomy adversely affected GFR. As intersegmental nephrotomy requires additional surgical manipulation and time, bisection nephrotomy is the technique of choice for nephrotomy in dogs. PMID- 12094356 TI - Healthy children 2010: implications for pediatric nursing practice. PMID- 12094355 TI - Effects of degree of acetabular rotation after triple pelvic osteotomy on the position of the femoral head in relationship to the acetabulum. AB - OBJECTIVE: To assess the effect of rotating the acetabulum 20 degrees and 30 degrees on percent coverage (PC) of the femoral head by the acetabulum and Norberg angle (NA) after triple pelvic osteotomy (TPO). STUDY DESIGN: Retrospective study. ANIMAL POPULATION: Fifty-six client-owned dogs. METHODS: PC and NA were measured from preoperative, immediate postoperative, first recheck, and second recheck radiographs to compare the effects of 20 degrees and 30 degrees of rotation of the acetabulum after TPO for treatment of hip dysplasia. RESULTS: Within the 20 degrees rotation group, PC and NA values were significantly greater at each subsequent evaluation time starting at the preoperative measurement except for NA between the first and second recheck times. Within the 30 degrees rotation group, PC and NA values were significantly greater at each subsequent evaluation time starting at the preoperative measurement except for PC and NA between the first and second recheck times. No significant differences between the 20 degrees and 30 degrees rotation groups were found at any corresponding times for either PC or NA. CONCLUSIONS: Acetabular rotation of 20 degrees provided the same degree of improvement in acetabular femoral head coverage as 30 degrees of rotation after TPO at the immediate postoperative, first recheck, and second recheck evaluation times. CLINICAL RELEVANCE: Based on PC and NA, 20 degrees rotation of the acetabulum provides as much benefit as 30 degrees of acetabular rotation when performing a TPO. PMID- 12094357 TI - Using the resiliency model to deliver culturally sensitive care to Chinese families. AB - In 1998, 4% of U.S. children were Asian/Pacific Islander; of these, 23% were Chinese. The incidence of infants with congenital heart defects (CHD) in Chinese children approximates that of Caucasian children in the United States. Parents of children with CHD are faced with caring for the children at home after surgery. Parents' roles and responsibilities expand significantly as their children return home with complex care needs. In pediatric nursing, we not only take care of the individual child but also the family as a whole. The family is the primary institution in society that preserves and transmits culture. Understanding Chinese culture and its association with health care practice helps nurses to establish culturally sensitive nursing interventions; develop appropriate discharge plans; decrease family stress; reduce future medical hospitalization costs; increase positive coping, adaptation, and family function; and improve health outcomes of the child and the family. The Resiliency Model was used to examine its applicability in caring for Chinese families in this paper. PMID- 12094358 TI - Patterns of family adaptation to childhood asthma. AB - This study examined patterns in adaptation among parents with a child who had moderate to severe persistent asthma. Specifically, we were interested in examining the differences in adaptation between mothers and fathers in which it was hypothesized that gender effects would be obtained in patterns of coping. Eighty-four parents participated in the study, representing 37 intact families in which both parents were present and 13 single-parent families. Within intact families, mothers exhibited greater efforts than fathers in coping patterns including strategies to acquire social support outside the family, enhance self worth, and decrease psychological tensions. When compared to mothers in single parent families, mothers within intact families had a greater tendency to use coping patterns related to family integration and cooperation. Such findings demonstrate a need for additional support for mothers in their role in caring for the chronically ill child. The implications of these findings for clinical practice are discussed. PMID- 12094359 TI - Maternal factors related to parenting young children with congenital heart disease. AB - The purpose of this study was to compare the early child-rearing practices between mothers of young children with congenital heart disease (CHD) and mothers of healthy children. In addition, maternal stress, parental developmental expectations, and the early behavioral and emotional development of their children were explored. Maccoby's (1992) socialization theory emphasizing the reciprocal nature of mother-child interactions provided the framework for this study. Findings from quantitative self-report measures and videotaped parent child interactions showed a remarkable similarity between mothers of children with CHD and mothers of healthy children. In contrast, qualitative data revealed important differences with mothers of CHD children reporting high levels of vigilance with their children. The important role of promoting the principle of normalization among mothers of children with CHD and ensuring a sufficient support system is discussed. PMID- 12094360 TI - Mother-toddler interaction and the development of self-regulation in a limit setting context. AB - The Prohibition Coding Scheme was used to longitudinally assess observations of maternal and toddler limit-setting interactions at 12, 24, and 36 months. The final sample consisted of 126 mother-toddler dyads. Toddler behavior in a limit setting context exemplified progressing skill in self-regulation from 12 to 36 months. Specifically, toddlers exhibited less persistence, increasing autonomy, and increasing ability to inhibit their behavior, with half of the toddlers classified as "autonomous-compliant" by 36 months of age. As a group, mothers also adapted their socializing behavior during toddlerhood, decreasing their use of directive structure and distractions, and using more cognitive strategies. A maternal "teaching-based", limit-setting pattern was associated with more competent toddler response-to-limits patterns at each age, and more mothers used a teaching-based limit-setting style by 36 months of age. These findings provide validity for the Prohibition Coding Scheme and extend our knowledge base for guiding parents in their socializing strategies relative to developing self regulation. Clinical implications and areas for further research are addressed. PMID- 12094361 TI - The care of my child with cancer: a new instrument to measure caregiving demand in parents of children with cancer. AB - The growing societal trend toward delivering more and more illness-related care in the home, driven both by family preferences and by mandates from third-party reimbursers, places additional responsibilities for increasingly complex caregiving on parents of children with serious illness. This article reports on the development and initial field test of The Care of My Child with Cancer, a caregiving demand instrument specific to the childhood cancer population. The instrument demonstrated strong internal consistency and test-retest reliability, and exploratory factor analysis provided initial evidence for the instrument's construct validity. The instrument will now be applied in a collaborative program of nursing research to further investigate caregiving demand and ultimately to develop nursing interventions to maximize medical and quality of life outcomes for children with cancer and their families. PMID- 12094362 TI - The impact of asthma risk factors on home management of childhood asthma. AB - There have been few studies of childhood asthma among families who live in nonmetropolitan settings. This work is part of the baseline assessment conducted before implementing a health education program to study the impact of asthma risk factors (gender, ethnicity, socioeconomic status, asthma severity) on home asthma management. Data analysis yielded no significant differences in home asthma management performed by parents or children with asthma based on the child's gender, ethnicity, asthma severity, or family socioeconomic status. Factors that define the child's experienced asthma pattern, such as activity limitations, number of allergens, and school absenteeism, were associated with the parent's work of asthma management. Trends in the data for the different ethnic and gender subgroups that have implications for clinical practice were identified. Future directions for research to address questions that emerged in this analysis are discussed. PMID- 12094363 TI - Green birthdays: reducing harmful exposures. PMID- 12094364 TI - Babies sleeping. PMID- 12094365 TI - Suspected child abuse: communicating with a child and her mother. AB - The purpose of this article is to explore and examine how the value judgments of the health professionals involved affected the interaction between themselves and a mother with her injured child who presented at an accident and emergency department. The interaction is depicted by means of a classroom role-play in which the "actors" are nurses. Analysis of the interaction demonstrates clearly how the perspectives of health professionals may be enshrouded by a miasma of suspicion. Indeed, by focusing on the actions of the mother rather than the needs of the child, suspicion gained momentum and finally entered the public domain. Encouraging health professionals to listen to what the child has to say goes some way to challenging the suspicion of abuse. Nonetheless, in the role-play analyzed, the mounting suspicion so antagonized the mother that an angry confrontation with the doctor ensued, the doctor being forced to make an embarrassing apology. Yet the interaction is so irredeemably prejudiced that it remains impossible either to confirm or reject the suspicion of abuse. Analysis of the interaction attempted to unravel some of the knots in which we, as health professionals, unfailingly tie ourselves when communicating with patients-knots of suspicion, labeling, conflict, embarrassment, and power that fail to anchor the humanity of "self" or others. PMID- 12094366 TI - Children's understanding of illness: students' assessments. PMID- 12094368 TI - Clinicopathologic correlations in gastrointestinal stromal tumors. PMID- 12094369 TI - Gastrointestinal stromal tumors: answers and questions. PMID- 12094370 TI - Diagnosis of gastrointestinal stromal tumors: A consensus approach. AB - As a result of major recent advances in understanding the biology of gastrointestinal stromal tumors (GISTs), specifically recognition of the central role of activating KIT mutations and associated KIT protein expression in these lesions, and the development of novel and effective therapy for GISTs using the receptor tyrosine kinase inhibitor STI-571, these tumors have become the focus of considerable attention by pathologists, clinicians, and patients. Stromal/mesenchymal tumors of the gastrointestinal tract have long been a source of confusion and controversy with regard to classification, line(s) of differentiation, and prognostication. Characterization of the KIT pathway and its phenotypic implications has helped to resolve some but not all of these issues. Given the now critical role of accurate and reproducible pathologic diagnosis in ensuring appropriate treatment for patients with GIST, the National Institutes of Health convened a GIST workshop in April 2001 with the goal of developing a consensus approach to diagnosis and morphologic prognostication. Key elements of the consensus, as described herein, are the defining role of KIT immunopositivity in diagnosis and a proposed scheme for estimating metastatic risk in these lesions, based on tumor size and mitotic count, recognizing that it is probably unwise to use the definitive term "benign" for any GIST, at least at the present time. PMID- 12094371 TI - Clinical management of gastrointestinal stromal tumors: before and after STI-571. AB - Gastrointestinal stromal tumor (GIST) is the most common mesenchymal neoplasm of the gastrointestinal tract. Until recently, surgery has been the only effective therapy for GIST. However, even after complete resection of tumor, many patients still eventually die of disease recurrence. Conventional chemotherapy and radiation therapy have been of limited value. Within the last few years, it was discovered that most GISTs have a gain-of-function mutation in the c-kit proto oncogene. This results in ligand-independent activation of the KIT receptor tyrosine kinase and an unopposed stimulus for cell growth. STI-571 is a small molecule that selectively inhibits the enzymatic activity of the ABL, platelet derived growth factor receptor, and KIT tyrosine kinases and the BCR-ABL fusion protein and is a landmark development in cancer therapy. Its clinical development marks a new era of rational and targeted molecular inhibition of cancer that emanates from direct collaborations between scientists and clinicians. It provides proof of the principle that a specific molecular inhibitor can drastically and selectively alter the survival of a neoplastic cell with a particular genetic aberration. The advent of STI-571 has markedly altered the clinical approach to GIST. It has proven to be effective in metastatic GIST and is also under investigation as a neoadjuvant and adjuvant therapy. PMID- 12094372 TI - Evaluation of malignancy and prognosis of gastrointestinal stromal tumors: a review. AB - This paper reviews data on the prognosis of gastrointestinal stromal tumors (GISTs). These tumors are specific KIT-expressing and KIT-signaling-driven mesenchymal tumors, many of which have KIT-activating mutations. GISTs occur in the entire gastrointestinal (GI) tract and may also arise from the omentum, mesenteries, and retroperitoneum. They range from small benign tumors to sarcomas at all sites of occurrence. A KIT tyrosine kinase inhibitor, STI-571 (imatinib [Gleevec]; Novartis, Basel, Switzerland), has recently shown promise in the treatment of metastatic GISTs. Understanding the natural history of GIST before introduction of STI-571 will help assess the impact and position of this new treatment. The frequency of benign versus malignant GISTs varies between sites. Benign GISTs outnumber malignant GISTs in the stomach, whereas malignant GISTs are more common in the intestines. Tumors that have metastasized at presentation have a very poor prognosis. Traditionally, the 3 key prognostic factors have been mitotic rate, tumor size, and site. Tumors that are small (< or =2 cm) and show mitotic activity not exceeding 5 mitoses per 50 high-power fields (HPFs) have an excellent prognosis, probably independent of site, although this has not been shown specifically for all sites. In the stomach, most epithelioid GISTs are benign, provided that mitotic counts do not exceed 5/50 HPFs. However, a small proportion of tumors apparently lacking mitotic activity do metastasize. Tumors with a mitotic rate >5/50 HPFs usually have a malignant behavior. The Ki67 index may help identify tumors with malignant potential, but large site-specific series are not yet available. Genetic markers, including DNA-copy number changes, telomerase activity, and KIT mutation status, may be useful in more accurately identifying tumors with malignant potential. PMID- 12094373 TI - Biology and genetic aspects of gastrointestinal stromal tumors: KIT activation and cytogenetic alterations. AB - Recent studies have done much to reveal the biological and genetic underpinnings of gastrointestinal stromal tumors (GISTs). Constitutive activation of the KIT receptor tyrosine kinase is a central pathogenetic event in most GISTs and generally results from oncogenic point mutations which can involve either extracellular or cytoplasmic domains of the receptor. Oncogenic mutations enable the KIT receptor to phosphorylate various substrate proteins, leading to activation of signal transduction cascades which regulate cell proliferation, apoptosis, chemotaxis, and adhesion. KIT mutations can be broadly assigned to 2 groups, those that involve the "regulatory" regions responsible for modulating KIT enzymatic activity and those that involve the enzymatic region itself. In vitro studies suggest that GISTs with regulatory-region KIT mutations are more likely to respond to STI-571 than are GISTs with enzymatic-region mutations. A minority of GISTs lack demonstrable KIT mutations, but KIT is nonetheless strongly activated. Such GISTs might contain KIT mutations which are not readily detected by conventional screening methods, or alternately, KIT might be activated by nonmutational mechanisms. Most GISTs have noncomplex cytogenetic profiles, often featuring deletions of chromosomes 14 and 22. Additional chromosomal aberrations are acquired as the GISTs progress to higher histologic grade. These cytogenetic aberrations are undoubtedly important in GIST pathogenesis, but currently they do not play a key role as diagnostic adjuncts. PMID- 12094374 TI - Expression of SSX genes in the neoplastic cells of Hodgkin's lymphoma. AB - The cancer/testis antigen (CTA) group of tumor-associated proteins have been reported to be expressed in various cancers and in adult testis but they are essentially not found in any other normal adult nonneoplastic tissues. Prompted by the frequent detection of SSX1 in a previous comprehensive expression profile of the Hodgkin's lymphoma (HL) cell line L428, we analyzed SSX expression by nonnested reverse-transcription polymerase chain reaction (RT-PCR) in 4 HL cell lines (L428, L540, HD-MY-Z, and KM-H2) and 32 tumor samples of HL. The cellular localization of SSX expression in the tumor samples was further analyzed by in situ hybridization (ISH). All 4 HL cell lines were positive by RT-PCR using SSX consensus primers. Using primers specific to individual SSX genes, all 4 cell lines expressed multiple SSX family members. Five tumor samples (15.6%) were positive by RT-PCR using SSX consensus primers and direct sequencing of the RT PCR products showed that 4 of 5 expressed more than 1 SSX family member. ISH confirmed that SSX expression originated in HL cells in all 5 RT-PCR-positive tumor samples. Furthermore, ISH demonstrated SSX-positive HL cells in 6 of 11 cases (55%) that were negative by RT-PCR. Our results suggest that members of the SSX family of CTA are expressed in most HL. This subset of HL may be a candidate for immunotherapy approaches directed at SSX proteins. PMID- 12094375 TI - Increasing expression of gastrointestinal phenotypes and p53 along with histologic progression of intraductal papillary neoplasia of the liver. AB - Intraductal papillary neoplasia of the liver (IPN-L) was recently proposed as the name for intraductal papillary proliferation of neoplastic biliary epithelium with a fine fibrovascular stalk resembling intraductal papillary mucinous neoplasm of the pancreas. We histochemically and immunohistochemically examined IPN-L alone or associated with hepatolithiasis, with an emphasis on the gastrointestinal metaplasia, nuclear p53 expression, and histologic progression. A total of 66 cases of IPN-L were divided into 4 groups: group 1, IPN-L with low grade dysplasia (13 cases); group 2, IPN-L with high-grade dysplasia (20 cases); group 3, IPN-L lined with carcinoma in situ and no or microinvasion (19 cases); and group 4, group 3 with distinct invasive carcinoma (14 cases). It is suggested that IPN-L progresses from group 1 to group 4. As controls, 20 cases of nonneoplastic intrahepatic large bile ducts and 17 cases of nonpapillary invasive intrahepatic cholangiocarcinoma (ICC) were used. Biliary epithelial hypersecretion of sialomucin rather than sulfomucin was prevalent in IPN-L, and this was associated with the progression of INP-L. Immunohistochemically, cytokeratin (CK) 20 and MUC2, a gastrointestinal marker, were expressed more frequently in IPN-L than in nonneoplastic bile ducts and nonpapillary ICC (P <0.01), and their incidence were significantly increased in parallel with the progression of IPN-L (P < 0.01). In contrast, expression of CK 7, a biliary marker, was decreased in IPN-L compared with nonpapillary ICC. Nuclear p53 immunostaining was detected in 30% of IPN-L as a whole and increased in tandem with the progression of IPN-L (P < 0.01). It is suggested that IPN-L forms a spectrum of biliary epithelial neoplasia with frequent gastrointestinal metaplasia, different from the usual nonpapillary ICC, and shows stepwise progression from the perspective of mucin profile, gastrointestinal metaplasia, and p53 nuclear expression. PMID- 12094376 TI - Prognostic significance of atypical papillary urothelial hyperplasia. AB - Typical papillary hyperplasia, a recently recognized precursor lesion to low grade papillary urothelial neoplasms, consists of undulating folds of cytologically benign urothelium. Well-developed, branching fibrovascular cores of a papillary neoplasm are not evident. We have noted lesions with the architectural pattern of papillary hyperplasia; however, the overlying urothelium demonstrated varying degrees of cytologic atypia. We identified 15 cases of atypical papillary hyperplasia (13 males, 2 females, age 55 to 92) with overlying urothelium showing cytologic atypia. Of these cases, 8 (53%) were received in consultation. Of the 15 cases, 8 exhibited overlying flat carcinoma in situ (CIS), 4 had overlying dysplasia, and 3 were transitional between papillary hyperplasia with atypia and the earliest lesions of papillary neoplasia. Of these cases, 5 patients had multiple specimens with atypical papillary hyperplasia (range, 2 to 8) over time. Concurrent to the diagnosis of atypical papillary hyperplasia, there were 25 different urothelial lesions: CIS (n = 11), papilloma (n = 1), papillary neoplasm of low malignant potential with CIS (n = 1), high grade papillary urothelial carcinoma (n = 10; 3 with CIS), small-cell carcinoma (n = 1), and infiltrating urothelial carcinoma (n = 1). Of 11 patients with known prior history, 2 had 12 prior urothelial neoplasms (9 low-grade papillary neoplasms, 2 papillary urothelial neoplasms of low malignant potential, and 1 high-grade papillary cancer). Of 10 patients with atypical papillary hyperplasia and a minimum of 1 year of follow-up, 9 had 19 recurrences: CIS (n = 4), papilloma (n = 1), papillary neoplasm of low malignant potential (n = 1), infiltrating urothelial carcinoma (n = 3; 1 with CIS), and high-grade papillary urothelial carcinoma (n = 10; 5 with invasion and 2 with CIS). Whether the papillary hyperplasia had overlying CIS or dysplasia did not affect the correlation with urothelial neoplasms. Immunohistochemical analysis of p53 and Ki 67 expression in 8 cases demonstrated overexpression of p53 (n = 2; 1 with overlying dysplasia and 1 with overlying CIS), and Ki-67 (n = 5; 2 with overlying dysplasia and 3 with overlying CIS). Taken together, these results suggest that atypical papillary hyperplasia is most frequently associated with CIS and high grade papillary cancer. In some cases, CIS or dysplasia may evolve into atypical papillary hyperplasia, with further progression to high-grade papillary cancer. This process may be analogous to papillary hyperplasia without cytologic atypia progressing to low-grade papillary urothelial neoplasms. PMID- 12094377 TI - Central zone histology of the prostate: a mimicker of high-grade prostatic intraepithelial neoplasia. AB - The central zone (CZ) is located at the base of the prostate adjacent to the seminal vesicles. Its histology as a potential mimicker of high-grade prostatic intraepithelial neoplasia (PIN) has not been formally studied. Three groups were evaluated. Group 1 comprised 30 consecutive radical prostatectomy specimens assessed for the extent of CZ and of Roman arch and/or cribriform formation in the CZ. Group 2 comprised 100 consecutive cases of nonconsult prostate needle biopsies, screened in a random blinded fashion to identify CZ histology and the specificity of its identification on biopsy. Group 3 comprised 34 consult cases (1984 to the present) with CZ histology on needle biopsy. For group 1, the average maximum diameter of CZ histology was 5 mm. Two cases (6.7%) did not contain the classic features of CZ histology. The average amount of cribriform and/or Roman arch formation in the areas with CZ histology was 16.5%. In group 2, 10% of prostate needle biopsy cases had CZ histology. Of these, 80% were located on biopsy specimens designated as the base of the prostate, 10% were located in the base and midportion of the prostate, and 10% were located in the midportion of the prostate. For group 3, CZ histology occupied on average 32% of the involved core. The 2 most common histologic features were eosinophilic cytoplasm (97%) and location at the end of a core (97%). Other features were Roman arch formation (59%), a prominent basal cell layer (32%), cribriform formation (26%), and associated thick muscle bundles typical of bladder neck (24%). On average, cribriform and/or Roman arch formation occupied 22% of the CZ area seen on biopsy. Twenty-six of the consult cases were sent in with preliminary outside diagnoses. Of these, 21 (81%) were either PIN or atypical: 11 (42%) high-grade PIN, 7 (27%) PIN, and 3 (12%) atypical glands. Our findings show that CZ histology is distinctive, as seen in radical prostatectomy specimens. Less frequently it is found on needle biopsy, where the presence of Roman arch and/or cribriform formation mimics PIN. Recognition of the distinctive features of CZ histology (i.e., tall columnar cells with eosinophilic cytoplasm, prominent basal cell layer, and lack of cytologic atypia) can help avoid a misdiagnosis of PIN or "atypia" on needle biopsy. PMID- 12094378 TI - Excessive survivin expression in thyroid lymphomas. AB - Thyroid lymphoma occurs most commonly in the thyroid glands with a background of Hashimoto's thyroiditis. Therefore, it is occasionally difficult to distinguish lymphoma from Hashimoto's thyroiditis because of some cellular and histologic similarities. We have examined whether survivin or human telomerase reverse transcriptase (hTERT) expression can differentiate between the 2 disorders. Surgically removed tissue samples from 6 patients with thyroid lymphoma and 6 patients with Hashimoto's thyroiditis were analyzed for mRNA levels of survivin and hTERT by real-time quantitative reverse-transcription polymerase chain reaction. Expression of survivin protein was examined by immunohistochemical stain using a polyclonal antibody. Survivin mRNA levels were greater in thyroid lymphoma than in Hashimoto's thyroiditis: 49.1 +/-36.4 versus 6.6 +/-2.7 pg/ng rRNA (mean +/- SD) (P <0.005). Immunohistochemical stain confirmed an abundance of survivin protein in lymphoid cells of thyroid lymphoma. The amount of hTERT mRNA did not differ in the 2 disorders. Our study shows that measuring survivin mRNA levels or immunohistochemistry of the protein expression can be useful to aid the diagnosis of thyroid lymphoma when histologic diagnosis is difficult. PMID- 12094379 TI - Clonality of oligoastrocytomas. AB - Oligoastrocytomas (OA) are mixed glial tumors that show morphologic features of both oligodendrogliomas and astrocytomas. The histogenesis of these tumors remains undefined. The aim of this study was to investigate the clonality of OA on the basis of tumor-dependent genetic alterations and tumor-independent X chromosome inactivation. We microdissected 11 biphasic OA and subjected the oligodendroglial and astrocytic components to allelic loss analysis of chromosomes 1p, 9p21, 10q, 13q, 17p, and 19q; TP53 immunohistochemical and mutation analyses; and X-linked HUMARA gene methylation study. On the basis of the genetic findings, we categorized these tumors into 3 groups. Group 1 consisted of 4 tumors that showed identical genetic aberrations in the 2 histologic elements, characterized by allelic loss on 1p and 19q. These results suggest that group 1 tumors are of monoclonal origin and share a precursor cell with oligodendrogliomas. Group 2 consisted of 5 tumors characterized by losses on 1p and 19q, with additional allelic losses on chromosomes 9p, 10q, 13q and/or 17p. Four of these tumors were of the anaplastic type. Thus, group 2 tumors may be regarded as advanced variants of group 1 OA with heterogeneous genetic changes during clonal expansion. The X-chromosome inactivation analysis confirmed the monoclonality of groups 1 and 2 OA. Group 3 consisted of two tumors that showed divergent allelic loss patterns in the 2 histologic components. Mutation and overexpression of TP53 were detectable in the astrocytic components only. These findings raise the possibility that group 3 tumors have a biclonal origin. In conclusion, our results suggest that OA are predominantly of monoclonal origin but that a small subset of tumors may be derived from different precursors. PMID- 12094380 TI - Nup88 (karyoporin) in human malignant neoplasms and dysplasias: correlations of immunostaining of tissue sections, cytologic smears, and immunoblot analysis. AB - Nuclear pore complexes (NPCs) are elaborate macromolecular structures that regulate the bidirectional nucleocytoplasmic traffic system. In vertebrate cells, NPCs include a family of 50 to 100 proteins termed nucleoporins (Nups). The 88-kD Nup has been found to be linked in a dynamic subcomplex with the oncogenic CAN/Nup214. Applying a polyclonal antiserum to Nup88 on paraffin sections, we found that it immunoreacts with numerous malignant neoplasms. All carcinomas reacted irrespective of site, type, or degree of differentiation; often, high grade carcinomas stained more strongly and extensively. Some sarcomas (e.g., fibrosarcomas, leiomyosarcomas, liposarcomas, and rhabdomyosarcomas) reacted intensely; melanomas, gliomas, mesotheliomas, and malignant lymphomas also stained. In situ carcinomas of the colon, stomach, breast, and prostate stained convincingly, as did in situ melanomas; some samples of fetal tissues also reacted. Cytologic smears of some of the aforementioned tumors also stained. In selected samples, enhanced immunostaining of tissue sections and cytologic smears correlated strongly and consistently with immunoblot data. Immunoblots of the same tumors with antibodies to 2 other Nups (Nup214 and Nup153) showed no comparable enhancement. Therefore, it seems that in some malignant tumors, Nup88 overexpression is not parallelled by an overexpression of other Nups. Benign tumors, hyperplasias, and normal tissues showed weak and sporadic staining or absence of staining; immunoblots of the same samples yielded weak signals. Occasional highly proliferative hyperplastic-reactive processes showed focal staining. Thus, our correlative histologic, cytologic, and molecular data indicate that Nup88 may be viewed as a potentially useful, broadly based histodiagnostic and molecular marker of many malignancies and premalignant dysplasias, and further suggest that in some malignant tumors, Nup88 may be selectively overexpressed as compared with other Nups. Thus, we propose that Nup88 be designated as karyoporin. PMID- 12094381 TI - Uroplakin-III to distinguish primary vulvar Paget disease from Paget disease secondary to urothelial carcinoma. AB - Paget disease of the vulva can be mimicked by several disease entities histopathologically, but most of these entities can be clinically distinguished from vulvar Paget disease. However, vulvar Paget disease is in itself a heterogeneous group of epithelial neoplasms that can be similar both clinically and histopathologically. The subtypes of vulvar Paget disease include primary Paget disease arising from a pluripotent stem cell within the epithelium of the vulva, and secondary Paget disease of the vulva. Secondary vulvar Paget disease results from spread of an internal malignancy, most commonly from an anorectal adenocarcinoma or urothelial carcinoma of the bladder or urethra, to the vulvar epithelium. We have recently proposed that these lesions be classified as primary (of cutaneous origin) or secondary (of extracutaneous origin). These subtypes can present similarly as eczematoid skin lesions and may appear similar on routine hematoxylin and eosin-stained slides. Immunohistochemical studies can help differentiate between them. Our current study includes 17 patients with a pathologic diagnosis of vulvar Paget disease. We performed a panel of immunohistochemical stains, including cytokeratin (CK) 7 and 20, carcinoembryonic antigen (CEA), gross cystic disease fluid protein-15 (GCDFP-15), and uroplakin III (UP-III). Of these 17 patients, 14 (80%) had primary intraepithelial cutaneous Paget disease, 13 without invasion and 1 with associated invasion. Three patients had urothelial carcinoma with spread to the vulva, manifesting as secondary vulvar Paget disease. Immunohistochemically, primary vulvar Paget disease is immunoreactive for CK 7 and GCDFP-15, but uncommonly for CK 20. Vulvar Paget disease secondary to anorectal carcinoma demonstrates CK 20 immunoreactivity but is usually nonreactive for CK 7 and consistently nonimmunoreactive for GCDFP-15. Vulvar Paget disease secondary to urothelial carcinoma is immunoreactive for CK 7 and CK 20 but nonimmunoreactive for GCDFP 15. In addition, we propose the use of a new, commercially available antibody, UP III, which is specific for urothelium and, in our experience, is immunoreactive in secondary vulvar Paget disease of urothelial origin. The distinction between these 3 types of Paget and Paget-like lesions is essential in that the specific diagnosis has a significant influence on current treatment. The difference in surgical approach to the subtypes of vulvar Paget disease justifies classifying them into distinct lesions, which may be aided by the use of immunohistochemistry, including UP-III. PMID- 12094382 TI - Vulvar Paget disease of urothelial origin: a report of three cases and a proposed classification of vulvar Paget disease. AB - Extramammary Paget disease is generally considered a distinct entity that can involve the genital tract skin and may be associated with underlying adenocarcinoma. Evidence is presented that vulvar Paget disease represents a heterogeneous group of epithelial neoplasms that can be similar both clinically and histopathologically. Three cases of vulvar Paget-like disease that were manifestations of urothelial carcinoma are investigated. Vulvar Paget disease can be classified based on the origin of the neoplastic Paget cells as either primary (of cutaneous origin) or secondary (of noncutaneous origin). Each classification has 3 subtypes: primary, intraepithelial cutaneous Paget disease of the usual type; intraepithelial cutaneous Paget disease with invasion, and intraepithelial cutaneous Paget disease as a manifestation of underlying skin appendage adenocarcinoma; secondary, Paget disease of anorectal origin, Paget disease of urothelial origin, and Paget disease of other origin. This subclassification is based on a review of the literature and the current study of 3 patients with Paget-like disease of urothelial neoplastic origin. The 3 subtypes of vulvar Paget disease studied here can present similarly as eczematoid skin or vulvar mucosal lesions and may appear similar on routine hematoxylin and eosin-stained slides. Immunohistochemical studies can be used to help differentiate them. The distinction between these 3 types of Paget-like lesions is essential in that the specific diagnosis has a significant influence on current treatment. The difference in surgical approach to the subtypes of vulvar Paget disease justifies classifying them into distinct lesions to avoid potential confusion and unnecessary surgery. PMID- 12094383 TI - Detection of molecular cytogenetic aberrations in langerhans cell histiocytosis of bone. AB - Langerhans cell histiocytosis (LCH) is a disorder of unknown etiology that gives rise to clonal expansion of Langerhans-like cells or their precursors. The molecular basis include aberrant expression of several adhesion molecules and elevated expression of p53, c-myc, and H-ras. To identify new locations of LCH related oncogenes or tumor-suppressor genes, we performed comparative genomic hybridization (CGH) and loss of heterozygosity (LOH) analysis on a series of 7 bone LCH lesions. Recurrent abnormalities were found by the CGH in all cases representing losses of DNA sequences on chromosomes 1p, 5, 6, 7, 9, 16, 17, and 22q. Gain of DNA copy number was seen on chromosomes 2q, 4q, and 12. The CGH data were supplemented by LOH analysis by means of 85 polymorphic microsatellite markers distributed along chromosomes 1, 7, 9, and 22. The highest frequency of LOH was found on 1p region in 3 of 7 informative cases and on chromosome 7 in 4 cases. Allelic loss was also detected on chromosomes 9 in 2 of 7 informative cases and on 22q in 1 of 7 cases. These results indicate that the deleted chromosomal segments may contain genes important in LCH initiation and progression. PMID- 12094384 TI - Kimura's disease with unusual eosinophilic epithelioid granulomatous reaction: a finding possibly related to eosinophil apoptosis. AB - We report and discuss a case of Kimura's disease with an unusual eosinophilic epithelioid granulomatous reaction. A 3-year-old Japanese boy with eosinophilia and a high concentration of IgE developed lymphadenopathy and multiple cervical masses. A lymph node biopsy demonstrated the infiltration of eosinophils in the stroma, which is consistent with the findings of Kimura's disease. Interestingly, a number of apoptotic eosinophils was detected in the infiltrating eosinophils. Multiple epithelioid granulomas with central eosinophilic abscesses and necrosis were also observed. Macrophages and giant cells had phagocytosed the apoptotic eosinophils at the edge of the granulomas. In situ terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) assay showed that the TUNEL-positive eosinophils were both in the macrophages and in the central eosinophilic abscesses of the granulomas. These findings suggest that the eosinophils had undergone an accelerated apoptosis in this case of Kimura's disease, and that the epithelioid granulomas were produced by phagocytosis of the apoptotic eosinophils by macrophages. PMID- 12094385 TI - Lipomedulloblastoma in a child: a controversial entity. AB - Lipomedulloblastoma is regarded as a distinct entity that occurs exclusively in adults and has a low proliferative potential and a favorable outcome. We describe a rare case of lipomedulloblastoma in a 6-year-old female child showing a high labeling index that needs documentation. The various hypotheses of adiposal change are discussed. PMID- 12094386 TI - Gastric carcinoma resembling pancreatic mixed acinar-endocrine carcinoma. AB - A case of gastric carcinoma resembling pancreatic mixed acinar-endocrine carcinoma of 77-year-old female is presented. This type of gastric tumor has not been previously reported. The endoscopic mucosal resection specimen of the fundus contained a 1.2 x 0.9 x 0.3 cm, well-circumscribed, tan, soft nodular tumor with protruded configuration with a central recess. Histologically, the tumor was confined to the mucosa and submucosa and was characterized by three growth patterns; acinar, solid, and glandular. The growth patterns were intermingled. The tumor cells in the acinar component had round nuclei with prominent nucleoli and diastase-resistant, periodic acid-Schiff-positive, eosinophilic cytoplasm. Immunohistochemically, the tumor cells were positive for CAM5.2, cytokeratin (CK) 7, CK 20, trypsin, lipase, alpha-1-antitrypsin, and alpha-1-antichymotrypsin. The tumor cells in the solid component were positive for Grimelius stain and chromogranin A. The findings indicated that the tumor showed acinar and endocrine differentiation. There was no heterotopic pancreas tissue in the specimen. The patient was well without tumor at the 7-month follow-up. It is important to know the existence of this type of gastric cancer and to not confuse it with a metastatic lesion of the pancreatic origin. PMID- 12094387 TI - Adefovir nephrotoxicity and mitochondrial DNA depletion. PMID- 12094388 TI - Low-grade myxoid renal epithelial neoplasms with distal nephron differentiation: a distinct clinicopathologic entity? PMID- 12094390 TI - [Reduction of cardiovascular morbidity and mortality by combined antihypertensive drug therapy in patients with type 2 diabetes mellitus]. AB - The coexistence of diabetes mellitus in hypertensive patients doubles the number of cardiovascular events (relative risk: 1.73-2.77) and cardiovascular mortality (relative risk: 2.25-3.66). Therapeutic interventions concentrating on elevated blood glucose alone for prevention of late complications were not effective in essential reduction of cardiovascular morbidity and mortality. For that reason therapies, focussing on other macrovascular risk factors, did increase significantly in the last decade. Strategies for reduction of the macrovascular risk include the aggressive treatment of hypertension. In the now published hypertension intervention studies in diabetic patients many different objectives were studied. As we already know, lowering of blood pressure does reduce cardiovascular risk. The optimal blood pressure threshold is not known yet, but of major interest. To find the most effective antihypertensive agent and the most effective combination therapy for diabetic patients, the most frequently used antihypertensive agents were compared with each other. Very interesting are the special effects of some substances, which exceed the lowering of blood pressure, like effects on the endothelinn or on coagulation disturbances. These protective and antiatherosclerotic effects could possibly get relevance even in normotensive patients. Concerning this special question, ACE-inhibitors and Angiotensin II receptor antagonists are getting more and more in the focus of interest and seem to be superior compared to other substances. Concluding the existing evidence from hypertension studies the following recommendations can be deduced: Diabetic patients with at least one additional cardiovascular risk factor should get an ACE-inhibitor in combination with other antihypertensive agents or as monotherapy. For combination therapy all the available antihypertensive agents are appropriate and can lower blood pressure adequately. PMID- 12094391 TI - [Intracoronary brachytherapy--an update]. AB - Restenosis following angioplasty represents a major clinical problem in the field of percutaneous coronary interventions. Intravascular brachytherapy reduces risk of restenosis following percutaneous interventions of native lesions and in-stent restenosis up to 50%-60%. This effect can be shown for at least one to two years. This novel therapeutic strategy is limited by a higher rate of target vessel reinterventions, and late coronary thrombosis, when platelet inhibiting drugs has been withdrawn or after implantation of multiple stents. Currently, intracoronary brachytherapy is mainly considered for treatment of in-stent-restenosis. PMID- 12094392 TI - [Cardiovascular manifestations in inflammatory rheumatic diseases]. AB - Inflammatory rheumatic diseases show an increased cardiovascular mortality. The reasons for this are unclear. Possible associations between atherosclerosis and autoimmune diseases like rheumatoid arthritis and systemic lupus erythematosus can be illustrated. PMID- 12094393 TI - Vascular endothelial growth factor (VEGF) in ankylosing spondylitis--a pilot study. AB - INTRODUCTION: Angiogenesis is important for the pathogenesis of chronic inflammatory diseases in joints. Inflammation itself may upregulate the expression of VEGF in rheumatic diseases. Angiogenesis may become a new target for therapeutic intervention in inflammatory joint disease. AIM OF THE STUDY: To examine plasma levels of VEGF in AS patients and to test a possible correlation with serological and/or clinical parameters. PATIENTS AND METHODS: Sixteen consecutive patients with definite AS were recruited from the Gasteiner Heilstollen Hospital and compared to eight healthy probands as controls. VEGF was determined in EDTA plasma samples by using an ELISA kit. Data are given as mean values (+/- SEM). The Spearman two-sided test was used to test possible correlations. RESULTS: EDTA-plasma levels of VEGF were 75.3 +/- 19.0 pg/ml, compared to 13.8 +/- 4.7 pg/ml measured in the control group (P = 0.001). A significant correlation was found between plasma VEGF of AS patients and the BASMI score (r = 0.665, P = 0.013). Whereas VEGF was elevated in patients without treatment or NSAIDs (88.9 +/- 24.2 pg/ml), lower levels up to 43.8 pg/ml were found in patients treated with corticosteroids (34.7 +/- 4.0 pg/ml, P = 0.039). CONCLUSIONS: Disease status of AS appears to be associated with elevated VEGF plasma levels. Whether this reflects inflammation or a truly angiogenic pathomechanism requires further investigation. PMID- 12094394 TI - [Effects of high iodine doses in parenteral roentgen contrast media administration on parameters of the pro-/antioxidant balance]. AB - The aim of this study was to investigate effects of high iodine doses given parenterally as contrast medium on parameters of lipid status and thyroid hormone status as well as on parameters of the pro/antioxidative balance of spa patients. 29 patients with a comparable indication who had to undergo an angiography were chosen. The blood parameters of these patients were determined before and after the angiographic treatment. No provable changes of the thyroid parameters f-T3, f T4 and TSH were found after the angiography. In case of enzyme activities, the protective enzymes SOD and GSHPX showed no changes, while the concentrations of peroxides and MDA were increased significantly. Corresponding to this, the total antioxidative status and the vitamin E level decreased significantly. Altogether these results stand for a moderate deterioration of the antioxidative protective potential by the highly iodine containing contrast medium. PMID- 12094395 TI - [Pericarditis as the initial manifestation of retroperitoneal fibrosis--a case report]. AB - HISTORY AND CLINICAL FINDINGS: A 70-year-old patient with invasive ductal breast cancer underwent conserving surgery of the right breast and right axillary dissection as well as postoperative irradiation therapy. Five months later, she presented with dyspnoea and progressive weakness. INVESTIGATIONS: Clinically, the patient showed anasarca and petechial hemorrhages, laboratory tests revealed thrombopenia, hepatic dysfunction, radiologic investigations showed enlargement of the liver and spleen, effusions of the pleura and pericardium, and ascite. Echocardiography showed pericardial effusion without cardiac tamponade. TREATMENT AND COURSE: Despite supportive therapy the patient's performance status deteriorated significantly, the diagnosis of the underlying disease could not be established, the patient died with the clinical signs of cardiovascular failure. Autopsy revealed progressive retroperitoneal fibrosis with systemic involvement of pleura, pericardium, epicardium, myocardium, lungs, and kidneys and pericarditis. Retrospectively clinical symptoms were interpreted as right heart insufficiency due to pericardial effusion. CONCLUSION: This case report reminds of occurrence of manyfold clinic manifestations of retroperitoneal fibrosis in dependence of particular organic involvement and that retroperitoneal fibrosis represents a differential diagnosis. PMID- 12094396 TI - ["Science between sponsorship and corruption"]. PMID- 12094397 TI - [Sponsorship, conflict of interest and corruption: promoting science in discussion of medical ethics]. AB - Even in health care systems corruption is an increasing problem. Since the German Anti-Corruption-Law (1997) the areas of financial support, sponsoring and corruption in medicine are analyzed much more intensively. This article sketches the crucial points of the debate on medical funding and shows the consequences of the new jurisdiction. Most important is the balance between a transparent documentation and a too-far-reaching bureaucracy, which hinders medical research. Finally, this article presents core principles as helpful rules in practice. PMID- 12094398 TI - [Research support and the pharmaceutical industry in Germany]. AB - Improved life expectancy in the industrial world is a consequence of modern drug development and clinical research. Drug development requires long term investments. The costs for drug development increased considerably, because of higher safety and effectiveness standards. New drugs and the improvement of treatment strategies increased the success-rate and, at the same time, reduced the costs. In oncology drug development Germany ranks only second in global competition. Accelerations in the administrative processes of the universities for clinical research are urgently needed to better compete internationally. Furthermore the legal requirements installed in recent years hampered industrial and clinical research. Politics must see to it that Germany does not fall behind other industrial countries even further in the field of drug research and development. PMID- 12094399 TI - [Aims and background of the anticorruption law]. AB - Corruption not only should be banned sociopolitically, it is a tort of law in Germany. Any employee in public duty has a special responsibility. That is the reason why the law does not tolerate any kind of corruption particularly in the public workplace. On trial the facts have to be established. A medical doctor is liable only if he abused his position to get an advantage. Doctors who are in charge of economic decisional power have to be aware that their responsibility for public money is important and that any personal advantage, which is linked to economic decisions will be problematic. Therefore the principles of transparency and rational economics have to be acknowledged. PMID- 12094400 TI - [Is research in Germany up for sale? Results of a study of Transparency International Germany]. AB - The intransparency of the complex Public Health System opens up the possibility for the abuse of all groups acting in this system. Some of them damage this system without hesitation even by corruption, and keep their focus on the personal interests instead of public interests. Particularly within research there are medical scientists and experts, who work and publish being dependant on grants from the pharmaceutical industries. PMID- 12094401 TI - [Consequences of the German anticorruption law for sponsorship at university hospitals]. AB - The cooperation between the pharmaceutical industry and clinical researchers has become an increasingly important public issue. Concerns are mainly focused around independence of clinical research and the dedication of clinical researches solely to the welfare of their patients. The declining public confidence in self regulation has finally resulted in new legislation that has major impact on clinical research in Germany particularly at academic centers. PMID- 12094402 TI - [Physicians--victims or promoters of corruption?]. AB - According to the media the recent physician bribery scandal in Germany draws ever further sets. The public prosecutor determines against hospital physicians and coworkers of a pharmaceutical firm. The suspicion: Physicians were recompensed for using up medicines particularly with pleasure trips. Which is qualified in Germany regularly as bribery and advantage grant as well as aid for tax evasion, is punishable in Austria as unfaithfulness, gift acceptance as well as bribery. The following contribution lights up--from Austrian view--the criminal page of the narrow burr between permitted sponsoring and undue corruption in the medicine. Bribery is globally punishable in Austria. Allowances to physicians can be for the payee in particular gift acceptance (section 153a StGB) or gift acceptance by leading employees of a public enterprise (section 305 StGB), for the giver in particular bribery (section 307 StGB). Occasional allowances, which are not located in connection to a concrete business, but only promoted the sympathetic consideration of the recipient, are not usually punishable. The punishing frameworks for offensces reach up to three years imprisonment. In addition still the absorption of enriching comes (section 20 StGB). PMID- 12094403 TI - Don't leave patient safety to the roll of the dice. PMID- 12094404 TI - How instruction and feedback can select the appropriate T helper response. AB - The decision of the immune system to trigger immune responses that are, respectively, induced by Th1 or Th2 effectors is a critical one, because it profoundly influences disease outcome. We have recently constructed a mathematical model of Th1-Th2-pathogen interactions that shows that the major decisional events can often be successfully determined by the intrinsic behaviour of the T helper system itself. For certain dangerous types of pathogens, however, which replicate rapidly or have developed strategies to evade the immune response, additional stimuli may be necessary. As a possible mechanism for the decision-making process innate immune recognition has been proposed. Here we present an enlarged version of our model, which incorporates signals created from the innate immune system after pathogen recognition. The model analysis suggests that there is fault-tolerance of the T helper system to incorrect Th1 signals. In the presence of incorrect Th1 stimuli an initial Th1 response is shifted to the correct Th2-dominated response owing to the intrinsic T helper dynamics. By contrast, according to our model there is no fault-tolerance for incorrect Th2 signals. In fact, if timing is unimportant then Th2 signals are superfluous since the intrinsic T helper dynamics provide an automatic switch to Th2 if Th1 effectors fail to control the pathogen. Th2 signals may, however, be required to accelerate the onset of the Th2 response. Additionally, we discuss the role of feedback where successful pathogen destruction leads to up-regulation of activation of the effective T helper type. As one possibility we examine the role of CpG motifs as indicators for successful pathogen destruction. Differences between instructive and feedback mechanisms are highlighted. PMID- 12094405 TI - Asymptotic analysis of noise sensitivity in a neuronal burster. AB - A combination of asymptotic approaches provides a new analysis of the effect of small noise on the bursting cycle of a neuronal burster of elliptic type (type III). The analysis is applied to a stochastic model of an excitable spine, with an activity-dependent stem conductance, that exhibits conditional burst dynamics. First, we give an asymptotic approximation to the probability density for the state of the system. This density is used to compute several quantities which describe the influence of the noise on the transition from the silent to the active phase. Second, we also use a multiscale method to provide a reduced system for analysing the effect of noise on the transition out of the active phase. The combination of these two approaches results in a new framework for a quantitative description of how noise shortens the burst cycle, which measures the significant influence of small noise. For the stochastic spine model, this study suggests that small amplitude noise can significantly influence the activity-dependent morphological plasticity of dendritic spines. The techniques used in this paper combine probabilistic and asymptotic methods, and have been generalized for other noisy nonlinear systems. PMID- 12094406 TI - Spatial effects in modeling pharmacokinetics of rapid action drugs. AB - We develop a model to describe the time course of plasma concentration of neuromuscular blocking agents used as anesthetics during surgery. This model, which overcomes the limitations of the classical compartment models routinely used in pharmacokinetics, incorporates spatial effects due to heterogeneity in the circulation: it takes the form of a dispersion equation on a circular domain, with a time-dependent leakage term. This term is fitted to the functional form desired once first-stage transients have died out. Comparisons are made with clinical data by adjusting three parameters in the model. PMID- 12094407 TI - A numerical approach to the study of spatial pattern formation in the ligaments of arcoid bivalves. AB - In this paper, we employ the novel application of a reaction-diffusion model on a growing domain to examine growth patterns of the ligaments of arcoid bivalves (marine molluscs) using realistic growth functions. Solving the equations via a novel use of the finite element method on a moving mesh, we show how a reaction diffusion model can mimic a number of different ligament growth patterns with modest changes in the parameters. Our results imply the existence of a common mode of ligament pattern formation throughout the Arcoida. Consequently, arcoids that share a particular pattern cannot be assumed, on this basis alone, to share an immediate common ancestry. Strikingly different patterns within the set can easily be generated by the same developmental program. We further show how the model can be used to make quantitatively testable predictions with biological implications. PMID- 12094408 TI - Motion of nanobeads proximate to plasma membranes during single particle tracking. AB - Drag and torque on nanobeads translating within the pericellular layer while attached to glycolipids of the plasma membrane are calculated by a novel hydrodynamic model. The model considers a bead that translates proximate to a rigid planar interface that separates two distinct Brinkman media. The hydrodynamic resistance is calculated numerically by a modified boundary integral equation formulation, where the pertinent boundary conditions result in a hybrid system of Fredholm integrals of the first and second kinds. The hydrodynamic resistance on the translating bead is calculated for different combinations of the Brinkman screening lengths in the two layers, and for different viscosity ratios. Depending on the bead-membrane separation and on the hydrodynamic properties of both the plasma membrane and the pericellular layer, the drag on the bead may be affected by the properties of the plasma membrane. The Stokes Einstein relation is applied for calculating the diffusivity of probes (colloidal gold nanobeads attached to glycolipids) in the plasma membrane. This approach provides an alternative way for the interpretation of in vitro observations during single particle tracking procedure, and predicts new properties of the plasma membrane structure. PMID- 12094409 TI - Quantitative effects of medium hardness and nutrient availability on the swarming motility of Serratia liquefaciens. AB - We report the first controlled measurements of expansion rates for swarming colonies of Serratia liquefaciens under different growth conditions, combined with qualitative observations of the organization of the colony into regions of differentiated cell types. Significantly, the results reveal that swarming colonies of S. liquefaciens can have an increasing expansion rate with time. We compare and contrast the expansion rate results with predictions from a recent mathematical model which coupled key hydrodynamical and biological mechanisms. Furthermore, we investigate whether the swarming colonies grow according to a power law or exponentially (for large times), as suggested by recent theoretical results. PMID- 12094410 TI - Topologically non-linked circular duplex DNA. AB - The discovery of circular DNA, over 30 years ago, introduced an element of uneasiness in what had been, up to that point, the almost picture-perfect story of the elucidation of the molecular biology of heredity. If DNA indeed has the Watson-Crick right-handed helical secondary structure, then in circular DNA, thousands, or perhaps even millions of twists must be removed in each generation, and re-wound in the next generation. Although enzyme systems adequate for this task have long since been found and characterized, there have nevertheless arisen a number of proposals for alternative DNA structures in which the strands are topologically non-linked, so that they might separate during replication without having to be unwound. These structures have generally been put forth as theory only, and have been largely unaccompanied by experimental evidence to support their applicability to native DNA from living systems. Recently, however, a report has emerged suggesting that it might be possible to separate, intact, the individual single-stranded circular half-chromosomes which constitute the double stranded circular chromosomes of certain plasmids. This would not be possible unless the chromosomes had one of the alternative, topologically non-linked structures. It is widely believed that after a half-century of worldwide DNA research, any significant change to the Watson-Crick structure is unlikely to stand up to scrutiny. Nevertheless, the present author has found that in many instances in which the behavior of circular duplex DNA is considered to be explicable only in terms of the topologically linked helical model, it is also possible to explain that same behavior in terms of a topologically non-linked model. It is necessary, in these instances, to make certain logical assumptions which cannot be conclusively proven at the present time. The author herein offers an example of one such instance, namely an examination of the behavior of circular duplex DNA in an alkaline titration experiment, where conformational changes in DNA are deduced from changes in its buoyant density at pH's between 7 and 14. These data have been explained in terms of topological linkage between the DNA strands, but they can also be explained without invoking any such topological linkage, provided that the above-mentioned logical assumptions can be accepted. The principles which emerge from this are applicable to other settings in which knowledge of the topology of DNA is critical to the understanding of observed phenomena. PMID- 12094411 TI - Complex discrete dynamics from simple continuous population models. AB - Nonoverlapping generations have been classically modelled as difference equations in order to account for the discrete nature of reproductive events. However, other events such as resource consumption or mortality are continuous and take place in the within-generation time. We have realistically assumed a hybrid ODE bidimensional model of resources and consumers with discrete events for reproduction. Numerical and analytical approaches showed that the resulting dynamics resembles a Ricker map, including the doubling route to chaos. Stochastic simulations with a handling-time parameter for indirect competition of juveniles may affect the qualitative behaviour of the model. PMID- 12094412 TI - [Individuals and innovative techniques, ethical reflections on the progress of technology in surgery]. PMID- 12094413 TI - [The Longo technique: should it be adopted?]. PMID- 12094414 TI - [Alithiasic cholecystitis in the adult: etiologies, diagnosis and treatment]. AB - Acalculous cholecystitis represents 2% to 14% of cholecystectomies performed for acute cholecystitis. Its main etiology is ischemia of the gallbladder wall, which mainly occurs in critically ill patients, particularly in case of cardiovascular previous disease or diabetes. Acalculous cholecystitis associated with VIH are rare and have a better prognosis. Other etiologies are exceptional. Diagnosis of acalculous cholecystitis is difficult, with a lack of specificity of abdominal ultrasound for the diagnosis of ischemic cholecystitis. In all cases, cholecystectomy is a definitive treatment allowing certain diagnosis. Percutaneous drainage must be reserved to patients whose general condition does not allow general anesthesia. Medical treatment alone is not indicated in acalculous cholecystitis. PMID- 12094415 TI - [Endoscopic harvesting of the latissimus dorsi flap in breast reconstructive surgery. Feasibility study and review of the literature]. AB - STUDY AIM: Breast reconstructive surgery with latissimus dorsi flap is routinely performed with a long dorsal scar ransom. To reduce the scar the authors propose an endoscopic technique to harvest the pure latissimus dorsal flap. We evaluate our first experience. PATIENTS AND METHODS: Between April 2001 and december 2001, 13 patients were operate using an endoscopic pure muscular latissimus dorsal flap for breast reconstruction surgery. RESULTS: Mean operative time is 116 min. No open conversion was necessary. One patient was transfused. The average lymphatic drainage was of 2520 ml with removal of the drainage on postoperative day 15. CONCLUSION: The endoscopic harvesting of the latissimus dorsi pure muscular flap, brings less scar than the open surgery. PMID- 12094416 TI - [IMesenteric and mesocolic cystic lymphangiomas. Diagnostic and therapeutic management]. AB - STUDY AIM: Study of clinical, diagnostic and therapeutic aspects of mesenteric and mesocolic cystic lymphangiomas. MATERIAL AND METHODS: 15 cases were retrospectively analysed: 5 adults (mean age 36.8 years, range 26 to 46) and 10 children (mean age 23 months, range 0 to 5 years). Diagnosis was prenatal in 1 case. Symptoms were: abdominal pain (80%), fever (20%), abdominal mass (46%), occlusive syndrome (33%), chylous ascitis 1 case. Tumours were mesenteric (86%) or mesocolic (13%). RESULTS: Complete resection was performed in 11 cases (including 10 bowel resections), incomplete resections in 3 and doxycycline sclerotherapy once. Mean follow-up is 5 years. One recurrence occurred 6 years after complete resection and 1 tumour increased after incomplete resection. Patient treated by sclerotherapy was non symptomatic with a 3.5 years follow-up after last injection. CONCLUSION: Mesenteric and mesocolic cystic lymphangiomas are congenital benign tumours. Complete resection should be performed whenever possible. Intracystic sclerotherapy with doxycyclin is possible for unresectable lymphangiomas. PMID- 12094417 TI - [Surgery for pulmonary hydatid cyst in Mali]. AB - INTRODUCTION: The pulmonary hydatid disease has not been the subject of many studies in Mali. OBJECTIVE: List the cases hydatid cyst of the lung operated on in Mali, in order to analyze their surgical aspects. METHODS AND PATIENTS: This is a retrospective study of consecutive and non-selected cases. From 1960 to 2000, eleven cases of the pulmonary hydatid disease were operated on in Mali. The earliest case dates back to 1968 and the most recent was in 2000. All of these patients were taken care of at the "Point G Hospital" in Bamako. RESULTS: Patients were aged between 4 and 42 years, with an estimated average at 20. The gender ratio (F/M) was 1.75. The circumstance leading to the discovery stems from 8 cases of cough. Seven patients had at least a sign of complicated cyst. Out of a total of 12 cysts, 9 were located to the right lung. The treatment of the cyst consisted in a kystectomy accompanied by an atypical resection of the parenchyma on 6 patients and a typical resection of the attacked lobe on 5 others. The operating effects were complicated in 5 cases. No mortality was reported. Anatomopathology confirmed pulmonary hydatidosis in all cases. CONCLUSION: Paradoxically, the frequency of pulmonary hydatidosis is low in Mali. Patients were consulted late. This explains in part the high level of lobectomy during surgical treatment. PMID- 12094418 TI - [Intensive care unit hospitalization after colorectal surgery]. AB - OBJECTIVE: Intensive care units (ICU) support critically ill patients during the perioperative period. Few studies exist focusing on ICU hospitalisation after colorectal surgery. The objective of the study was to 1) detect predictive factors of mortality and length of stay in ICU after colorectal procedures, and 2) compare the autonomy status of the patients before and 30 days after their ICU stay. PATIENTS AND METHODS: This study followed a prospective non randomized cohort in our colorectal surgery unit. During a period of one year (January 1st to December 31th, 2000) 351 colorectal procedures were performed and 54 patients were admitted to ICU after surgery. For each patient, 37 parameters were collected on a standardized register. Predictive factors of mortality (30 days after the procedure) and ICU stay (up to 3 days) were studied by univariate and multivariate statistical analysis. Self autonomy before surgery and 30 days after was also investigated. RESULTS: "Multiple-intervention" was the only independent factor influencing mortality. Both "low autonomy status before surgery" and "pulmonary comorbidity" increased the length of stay. Regarding the 48 survivors, 45 (94%) recovered the same autonomy index as in the preoperative period 30 days after the procedure. CONCLUSION: This study highlights the poor predictive factors influencing mortality during or after ICU stay following colorectal surgery, and emphasizes two preoperative parameters increasing the length of stay up to 3 days. This should guide the informations given to the patients families. Finally, this study confirms the good quality of self-sufficiency after ICU stay even for a long time (over 3 days). PMID- 12094419 TI - [Does the size of pheochromocytoma influence the results of its laparoscopic excision?]. AB - BACKGROUND: The tumor size is considered as a limitation for laparoscopic adrenalectomy. The aim of this study was to assess diagnostic characteristics, hemodynamic modifications, and outcome of intraperitoneal laparoscopic adrenalectomy according to the size of pheochromocytoma. METHODS: Retrospective study from January 1997 to December 2000. Results were evaluated according to the size of pathologic study (< or > or = 5 cm). RESULTS: Forty two patients underwent laparoscopic adrenalectomy during this period. Among them, 11 (26%) were operated on for adrenal pheochromocytoma (6 patients < 5 cm and 5 patients > or = 5 cm). Preoperative systolic hypertension was 138 mmHg and 178 mmHg respectively (p = 0.01). Urinary metanephrine/normetanephrine rate was 1.259 and 0.268 respectively (p = 0.08). Capsular tumor effraction rate was 27%. Tumor size was 37% larger than that estimated by CT scan (24% versus 52%). Mean hospital stay was 10 days and 8 days respectively. Morbidity and mortality were 18% and 0%. All these criteria were not statistically significant among the two groups of patients. CONCLUSIONS: Pheochromocytoma size > or = 5 cm do not modify outcome of patients but is responsible for preferential noradrenaline secretion and stronger hemodynamic modifications. Tumor fragmentation rate and CT scan size underestimation seem important in this group of patients. These characteristics have to be integrated to improve laparoscopic adrenalectomy outcome of patients with pheochromocytoma > or = 5 cm. PMID- 12094420 TI - [Adrenocortical carcinoma: prognostic factors for local recurrence and indications for reoperation. A report on a series of 22 patients]. AB - STUDY AIMS: The aim of this retrospective study was to identify prognostic factors on local recurrence in patients with adrenocortical carcinoma and to assess the effect of reoperations. METHODS: From 1985 to 2001, 22 patients were operated for adrenocortical carcinoma. We evaluated the correlation between actuarial survival without local recurrence and tumor staging, Weiss criteria, mitotic index, extensive resection and Op'DDD therapy by univariate analysis. Then we evaluated the effect of reoperations on survival. RESULTS: Local recurrence was observed in 7 patients and the 5-years actuarial survival without local recurrence was 50%. Tumor stage (I et II versus III et IV), Weiss criteria (< or = 6 criteria versus > 6) and mitotic index (< or = 20 mitoses/50 HPF versus > 20) affected survival without local recurrence. Extensive resection could reduce local recurrence rate. Op'DDD therapy was ineffective in prolonging survival without local recurrence. Four patients underwent repeat resections: 2 patients were still alive with disease free, 54 and 8 months after reoperations and 2 patients died, 19 and 25 months after reoperations. Three patients did not have repeat resection. They rapidly died within 8 months. CONCLUSION: Pathologic features (tumor staging and mitotic index) affected local recurrence prognostic. But extensive resection to adjacent organs could facilitate complete resection tumor and reduce the local recurrence rate. Complete repeat resection of local recurrence can improve survival. The disabling effects of Op'DDD were important and its efficacy was not clear. PMID- 12094422 TI - [Mesenteric venous thrombosis complicating acute appendicitis]. AB - Common primary surgical sources of thrombophlebitis of the mesenteric vein are diverticulitis and appendicitis. This is an acute ascending infection with septic thrombophlebitis. C.T. imaging can diagnose this complication at an early stage. Broad spectrum antibiotic therapy and heparin should be started. Surgery is performed electively to eradicate the primary inflammatory process. Early detections of septic ascending pylephlebitis and adequate treatment have decreased the mortality rate. We report a clinical case of thrombophlebitis of the mesenteric vein in acute appendicitis. PMID- 12094421 TI - [Spontaneous cholecystocutaneous fistula: natural history of biliary cholecystitis]. AB - Spontaneous cholecystocutaneous fistula is a rare complication of gallbladder empyema without treatment. The incidence has further decreased with the advent of resuscitation and surgical treatment. These fistulas are like a cholecystostomy. The diagnostic is confirmed with a fistulogram which allows visualization of the fistula tract. Traditional treatment has consisted of broad-spectrum antibiotics administration, cholecystectomy and fistula drainage. One-stage treatment is the preferred option. Laparoscopic approach appears feasible even the conversion rate is high. PMID- 12094423 TI - [Tuberculosis of the thyroid. Diagnosis and treatment]. AB - Thyroid tuberculosis is rare. We present the case of a 25 year-old woman with tuberculosis of the thyroid. Although seldom observed, tuberculosis should be considered in the differential diagnosis of nodular lesions of the thyroid. Diagnosis is made by histological examination and demonstration of the tubercle bacilli from biopsy or aspiration specimen. The efficacy of fine-needle aspiration cytology in diagnosis of tuberculosis of the thyroid is proved. Administration of antituberculous drugs is considered as the treatment of choice. Abscess drainage is sufficient. In rare cases surgery is necessary. The prognosis is good. PMID- 12094424 TI - Radiation-induced angiosarcoma of the breast: case report and self-criticism of therapeutic approach. AB - Angiosarcoma (AS) of the breast is a rare and highly aggressive vascular cancer. It presents as a primitive or radioinduced form. The case of a 46-year-old woman who underwent quadrantectomy of the breast plus axillary lymph node dissection and radiotherapy postoperatively (QUART) for ductal infiltrant carcinoma is reported in the following. Ten years later, the patient underwent mastectomy with immediate reconstruction, for local recurrence that was diagnosed as an AS of the breast at final pathological examination. She did not receive any adjuvant treatment due to local post-operative complications related to breast reconstruction. We criticize our therapeutic approach and we recommend more attention about local recurrence suggesting that tru-cut needle biopsy of local recurrence of the breast after QUART, should be the correct diagnostic approach. PMID- 12094426 TI - [Pierre Marion, 1914-2000. Past president of the National Academy of Surgery. An illustrious figure in the city of Lyons]. AB - The surgical work of Pierre Marion extends over a period of 40 years. It takes part in the history of portal hypertension surgery. Pierre Marion has been in France one of the major craftsmen of heart surgery with Paul Santy. In 1952, he performed the first pulmonary-embolectomy with success and later devoted his time to the development of valves and of an artificial heart. The second part of his life was dedicated to the direction of the Bullukian Foundation, founded in 1983, with the goal of helping medical research, supporting the armenian community in our country and promoting literary, artistic and theatrical creation in Lyons and its surroundings. Pierre Marion has written two remarquable books "Afin que batte le coeur", a history of the cardiac surgery epic and "Le genial bonhomme Ampere", consecrated to the life of this great scholar. PMID- 12094425 TI - [The difficult rectal stump]. AB - During proctectomy for rectal cancer, section and closure of the rectal stump with TA55 before stapled low colorectal anastomosis could be very difficult, especially in obese patient. In this situation, others solutions includes use of TA30 or Roticulator, closure of rectal stump by suture, use of laparoscopic endoGIA. If no device can be used, manuel colo-anal anastomosis can be performed after rectal version, instead of stapled very low colorectal anastomosis. PMID- 12094427 TI - [Arguments for surgical resection of hepatocellular carcinoma greater than 20 cm]. PMID- 12094428 TI - [Cavitary lung cancer and FDG-PET]. PMID- 12094429 TI - [Hepatic encephalopathy]. AB - Hepatic encephalopathy (HE) is a neuropsychiatric syndrome which develops during chronic or acute liver disease. It is functional in nature and potentially reversible and symptoms range from subtle personality changes to deep coma. Diagnosis of manifest HE is made on a clinical basis, whereas psychometric tests are required to diagnose subclinical HE (SHE). Paper-pencil tests are frequently used for diagnosing SHE, but they may be inferior to measurements of critical flicker frequency, which pick minimal and low grade manifest HE as a continuum. Pathogenetically, HE is seen as clinical manifestation of low grade chronic cerebral edema, which is accompanied by alterations in glioneuronal communication. Different factors such as ammonia, inflammatory cytokines, benzodiazepines and electrolyte imbalances may precipitate or aggravate glia edema, thereby explaining precipitation of HE episodes by a variety of unrelated factors. Recognition and rigorous treatment of these precipitating factors is the most important measure in HE therapy, which may be augmented by dietary and medical approaches. PMID- 12094430 TI - [Diagnosis of B and C viral hepatitis: new developments and relevance for general practice]. AB - Diagnosis of hepatitis B and hepatitis C viral infections can be achieved by highly specific and sensitive primary serological screening assays. Still more costly amplification systems on the qualitative and quantitative detection of HBV DNA and HCV-RNA are only used for answering special clinical questions. They are relevant to indicate antiviral therapy or to control the outcome of treatment. They are rarely necessary for diagnostic purposes as it may happen in immunosuppressed persons. While in hepatitis B activities of aminotransferases and liver histology usually present a good correlation this is not dependably seen in hepatitis C. Histologic evaluation still appears the only reliable diagnostic procedure for determining inflammatory activity and fibrosis progression in hepatitis C. Liver biopsy therefore is considered mandatory prior to initiation of treatment especially since only patients suffering from severe disease with progression to liver cirrhosis may benefit from today's standard treatment procedures. PMID- 12094431 TI - [Therapy of chronic hepatitis B infection]. AB - Around 350 million people worldwide are chronically infected with the hepatitis B virus. In adults around 5% and during perinatal contact around 100% of the infections become chronic. The main complications of chronic HBV infection are liver cirrhosis and hepatocellular carcinoma. Therefore after chronic HBV infection has been diagnosed early therapy has to be considered. At present two drugs are available: Interferon-alpha and lamivudine. Therapy with Interferon alpha is successful in 30-40% of the patients. In contrast, the initial response rate during lamivudine therapy is very good. However, in many of the patients mutant HBV strains are selected. Therefore in each patient it has to be considered individually which of the two drugs should be used. In this review these questions will be addressed and a special focus will be the use of lamivudine. Additionally future treatment options will be discussed. PMID- 12094432 TI - [Therapy of hepatitis C]. AB - Hepatitis C virus (HCV) infection is a leading cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma worldwide. Here, we will briefly review current and evolving therapies for chronic hepatitis C. Standard therapy with (pegylated) interferon-alpha and ribavirin achieves sustained response rates of up to about 40% in genotype 1- and about 80% in genotype 2- and 3-infected patients. Recent progress in the molecular virology and pathogenesis of hepatitis C has allowed the identification of novel antiviral targets and therapeutic strategies. These will likely complement existing therapeutic modalities in the near future. PMID- 12094434 TI - [Value of laparoscopy and biopsy]. AB - A liver biopsy can provide important information regarding staging, grading, prognosis and management of patients with chronic liver disease including chronic viral hepatitis. A video-minilaparoscopy is the most accurate diagnostic tool to define liver cirrhosis or liver fibrosis, respectively. PMID- 12094433 TI - [Therapy of hepatitis B and C. Problem situations in chronic HBV and HCV infection]. AB - In the most recent years, therapy of chronic hepatitis has been improved. Nonresponder to an initial treatment is the predominant group of difficult-to treat patients. Nonresponse may be due to host specific factors and/or virus specific properties. It is essential to reconsider the indication for treatment in respect to the presumed beneficial effect before initiating therapy or retreatment in these patients. Optimally, only those patients should be treated who are at significant risk for progression of liver disease. PMID- 12094435 TI - Fundamentally misunderstanding visual perception. Adults' belief in visual emissions. AB - The authors reviewed research about a profound misconception that is present among college students, namely, the belief that the process of vision includes emanations from the eyes, an idea that is consistent with the extramission theory of perception, which was originally professed by early Greek philosophers and which persisted in scholarly circles for centuries. The authors document the strength and breadth of this phenomenon and the object failure of traditional educational techniques to overcome this belief, and they reveal that students are leaving psychology courses with a flawed understanding of one of the most studied processes in the history of psychology--visual perception. Some suggestions are offered for overcoming this misconception in traditional college classroom settings. PMID- 12094436 TI - Presidential reflections. Past and future. AB - Key federal health initiatives and psychology's participation in public policy are briefly reviewed. There has long been agreement that psychologists have a responsibility to affirmatively address society's pressing needs, not only in health care but in other areas that affect the well being of society. The behavioral sciences are key to the United States' science and health policy in the 21st century. The unanswered question is, Has the field matured sufficiently to establish its own programmatic agenda via the public policy process? Community health, lifestyle choices, psychologically managed pharmacological treatment, and the use of telecommunications technology in education and treatment are areas in which psychology can contribute a great deal to the development of a healthier society. PMID- 12094437 TI - Psychology in the public service. AB - Philip G. Zimbardo outlines the challenges and opportunities he faces as the American Psychological Association's (APA's) 110th president. This article expands on remarks made in his introduction to Patrick H. DeLeon's presidential address at the APA's 2001 annual convention in San Francisco, California. Appearing now, mid-term in his presidency, that vision is a working blueprint of his activities and what he hopes to accomplish in his remaining tenure: enhancing psychologists' pride in psychology; developing more productive relationships with all media as gatekeepers to the public; publishing the standard high school psychology textbook; developing a compendium of all psychological research that illustrates how psychologists have made a significant difference in improving various aspects of the quality of life of individuals, groups, communities, and the United States; and encouraging greater unity of purpose and respect among psychologists across their many diverse domains and specialties. PMID- 12094439 TI - Ambivalent sexism inventory: where's the ambivalence? PMID- 12094440 TI - Maybe men and women are different. PMID- 12094441 TI - Ambivalent responses. PMID- 12094442 TI - Condom use reduces risk. PMID- 12094443 TI - Media violence: miscast causality. PMID- 12094444 TI - On media violence: whose facts? Whose misinformation? PMID- 12094445 TI - Media violence and the American public revisited. PMID- 12094446 TI - The value of national standards. PMID- 12094447 TI - Toward an integrative curriculum. PMID- 12094448 TI - What is a psychologist? PMID- 12094449 TI - The nonexistent common core. PMID- 12094450 TI - The struggle for a core curriculum. PMID- 12094451 TI - Revisiting psychology's core curriculum. PMID- 12094452 TI - Contributions of connectionism to postmodern psychology. PMID- 12094453 TI - The deconstruction of constructivism. PMID- 12094454 TI - Truth, method, and postmodern psychology. PMID- 12094455 TI - The dead end of postmodernism. PMID- 12094457 TI - Postmodern psychology and Africa. PMID- 12094456 TI - Attribution errors in the postmodern landscape. PMID- 12094458 TI - Postmodern parlor games. PMID- 12094460 TI - Psychological nescience in a postmodern context. PMID- 12094459 TI - More science, not less. PMID- 12094461 TI - Psychological science: to conserve or create? PMID- 12094462 TI - [Medical ethics and economics in health care: an irreconcilable contradiction?]. PMID- 12094463 TI - [Organ transplantation and insurability]. AB - Major organ transplant today is to be regarded as an established therapeutic procedure for treating irreversible organ failure. The creation of the necessary technological, immunological and ethical foundations in the 20th century brought about a true breakthrough in transplantation medicine. The presentation outlines various clinical pictures that can lead to chronic organ failure based on examples of heart, liver and kidney diseases. The long-term survival rates of patients/transplants are outlined and a comparison of international statistics drawn. Different problems that can arise in connection with a transplant, e.g. rejection reactions or transplant arteriosclerosis, are also broached. The heterogeneous risk profiles of the immunosuppressant drugs that the transplant recipient must take for the rest of his life are also discussed at length. There is no doubt about the fact that major organ transplants constitute a challenge to insurance medicine. The extreme extramortality of transplant recipients in the initial postoperative phase, makes immediate acceptance of transplant recipient applicants for life insurance prohibitive. It is, however, possible today to accept liver or kidney transplant recipients who have experienced a favourable post-transplant course, with adequate postponement. Finally, the presentation touches upon the different focuses of science's attempts to counteract a growing shortage of organs. PMID- 12094464 TI - [Prevention and rehabilitation services in private personal insurance]. AB - Prevention and rehabilitation services within the area of private insurances concerning the inability to work lead to an increased customer orientation, quality assurance and to the reduction of costs. With regard to prevention, different intervention possibilities are pointed out in order to reduce the risk of the event of a performance case. In the case of such the rehabilitation and vocational integration can be optimized by the application of a described procedure. An important component of the procedure represents the analysis of the vocational activity in order to identify the individuality of requirements and thus leading finally to an optimal fit to the abilities of the policyholder. PMID- 12094465 TI - [General medicine malpractice cases]. AB - During the last five years legal proceedings about alleged treatment mistakes in Germany more than doubled. Using a standardized questionnaire about legal proceedings in general medicine, involving liability, an anonymous survey with the members of the workgroup law medicine of the Deutsche Anwaltsverein (DAV) was carried out. The questions included among other things the number and reasons of legal proceedings involving liability. Of 322 questioned lawyers who focused on medicine law 122 (38%) answered. 69.9% of the lawyers think poor information is the main reason for legal proceedings involving liability in general medicine. Three disease groups were mentioned more frequently: diseases of the digestive system (22 mentions), diseases of the circulatory system (21) and diseases of the muscles, skeleton and connective tissue (15). 40 mentions of injections as treatment mistakes build the most frequent therapeutic reason for legal proceedings involving liability. Most of the lawyers think that guidelines do not reduce legal proceedings involving liability. The most common reasons for legal proceedings involving liability such as poor information and insufficient medical examination may point out that the budgetary standards for consultation cannot guarantee enough time for firstly giving a sufficient individual information to the patient and secondly for developing a decision satisfactory for both sides. PMID- 12094466 TI - [Illness related early pensioning of high school teachers]. AB - The problem of premature unfitness for work of (grammar school) teachers is of great social and social-medical interest. The possible causes and suitable measures of intervention remain a subject of controversy. The aim of this study was, on the basis of a random sample of a large number of cases, to gain a differentiated overview of the extent and type of early retirement due to illness of grammar school teachers, in order to be able to develop evidence-based strategies for prevention and intervention. In a prospective evaluation, all the assessments of illness-related unfitness for work drawn up for civil servants and teachers between 1.1.1996 and 31.12.1999 in Bavaria were first of all systematically evaluated. In addition, from the total collective a subgroup was formed of grammar school teachers and this was analysed separately. The answers given in a standardized, anonymous questionnaire provided the database. The evaluation included e.g. socio-demographic and occupational factors, the morbidity spectrum, medical judgement, medical rehabilitation and evaluation of performance. The medical diagnoses of the main and accompanying illnesses were classified according to ICD 10 in organ-related groups. Evaluation was carried out by means of descriptive statistics. Of the 655 grammar school teachers assessed, 65% (n = 429) were men, 35% (n = 226) women. The median age for male teachers was 58 years old (range: 30-64 years old), for female teachers 55 years old (range: 32-63 years old). Previous conflicts at the school workplace were reported in 12% of the cases. Among the main medical diagnoses, psychic/psychosomatic complaints (41%) predominated over muscular/skeletal illnesses (14%) and cardio-vascular diseases (12%). 80% of those examined were judged unfit for work. 56% of the grammar school teachers had participated in at least one medical rehabilitation measure before the proceedings for determining unfitness for work were instigated. Psychic and psychosomatic illnesses took first place among the decisive complaints leading to early retirement, and made up 45% of these. Further differentiation revealed the predominance of depressive illnesses and exhaustion (burnout). The main somatic complaints were far less common: muscular/skeletal illnesses (13%), cardio-vascular diseases (12%) and malignant growths (8%). The performance of 70% of the grammar school teachers judged unfit for work was regarded as so severely reduced that even activity outside of the teaching profession was no longer feasible. In this first representative evaluation of illness-related early retirement in grammar school teachers, the high prevalence and great social-medical importance of psychic and psychosomatic illnesses is noteworthy. In Bavaria, almost every second grammar school teacher therefore becomes unfit for work as a result of such complaints long before they reach the normal retiring age. From a social-medical and occupational-medical point of view, there is urgent need for further research into potential illness-inducing occupational and non-occupational stress factors and the development and implementation of effective and efficient strategies for prevention and intervention. PMID- 12094468 TI - [Musculoskeletal diseases and product warnings in product information provided by the computer industry]. PMID- 12094467 TI - [Communication center in public health]. AB - The Communications Center's portfolio covers areas such as marketing, contacts, distribution of information, sales activities and collection of bills by telephone (encashment). A special emphasis is Customer Care Management (Customer Relationship Management) to the patient and his caregivers (relatives), the customers, especially the physicians who send their patients to the hospital and the hospital doctor. By providing communication centers, the hospital would be able to improve the communication with the G.P.s, and identify the wishes and requirements more accurately and easily from the beginning. Dealing effectively with information and communication is already also of special importance for hospital doctors today. One can assume that the demands on doctors in this respect will become even more complex in the future. Doctors who are involved in scientific research are of course fully aware of the growing importance of the Internet with its new information and communication channels. Therefore analysing the current situation, the demands on a future information management system can be formulated: A system that will help doctors to avoid dealing with little goal oriented information and thus setting up effective communication channels; an information system which is multi-media oriented towards the interests and needs of the patients and patient's relatives and which is further developed continually and directly by those involved. PMID- 12094469 TI - [Occupational disability insurance--evaluating the work capacity in beauty culture regarding Meyerding II L4/5 Spondylolisthesis]. PMID- 12094470 TI - [Comment on J. Fritze: Stroke management in Germany]. PMID- 12094471 TI - [Comment on J. Fritze: Stroke management in Germany]. PMID- 12094472 TI - Basic principles of flow cytometry. AB - Although the basic principles of flow cytometry have changed little in the past quarter century, the applications of this technology have evolved substantially. As in the past, cytometers interrogate individual cells or particles in a stream with a laser as the cells move past a set of stationary detectors. Increasingly, more colors of fluorescence are being detected by cytometers, faster analysis and sorting rates are becoming possible, cytometers capable of multidirectional sorting are being marketed, and more reagents are becoming available for a wide variety of applications. Furthermore, flow cytometry has not stopped evolving. The development of narrow spectrum flourescent probes, the integration of molecular biologic techniques with flow cytometry, and the evaluation of cell free markers such as cytokines will be key components in the continuing evolution of flow cytometry. PMID- 12094474 TI - Acute myeloid leukemia and related conditions. AB - Flow cytometry is the primary tool for phenotyping leukemias and related conditions. With the ever increasing numbers of antibodies commercially available, the ability to study and understand leukemias, myelodysplastic syndromes, and the myelodysplastic/myeloproliferative diseases is improving. Although the data generated by flow cytometry are not comprehensive enough to completely subtype leukemias or myelodysplastic syndromes into their myriad divisions, this wealth of information does provide phenotype, reproducible enumeration of blasts, certain prognostic information, and it can reveal the presence of cell populations with aberrant antigen profiles. An important drawback to flow cytometry as it is performed today is the inability to look at the cells that mark with the antibody panels used. As classifications of leukemia and related conditions evolve, flow cytometry continues to answer many of the questions asked and to provide critical information reliably and quickly. PMID- 12094473 TI - Immunophenotypic analysis of acute lymphocytic leukemia. AB - Acute lymphoblastic leukemia (ALL) is one of the most common hematologic malignancies. Flow cytometry is an integral part of ALL diagnosis and also provides significant patient prognostic information. This article is a practical review of the basic principles of the flow cytometric evaluation of acute leukemias, the interpretation of flow cytometric data, and the management of practical problems such as aberrant antigen, hematogones, bone marrow regeneration, and minimal residual disease. PMID- 12094475 TI - B-cell lymphoproliferative disorders. AB - The differential diagnosis of the B-cell lymphoproliferative disorders can sometimes by tricky. When the morphology and flow cytometric studies are classic, the case should not present a diagnostic challenge. However, when the lesion does not read the book, one must take into consideration the morphology/cytology, flow cytometry, and clinical history. Integrating all three of these will often lead to the correct diagnosis; however, there will be occasional cases where the best that a pathologist can say is "B-cell lymphoproliferative disorder". In those instances, one should not overcall something, but rather should state what the problems are, and what are most likely to be the diagnostic possibilities. PMID- 12094476 TI - Monitoring of cellular resistance to cancer chemotherapy. AB - Cellular resistance to a broad spectrum of natural products used as antitumor drugs is believed to be a major cause for the failure of chemotherapy. Flow cytometry has been used for monitoring the expression of drug resistance markers, determining accumulation of fluorescent drugs, and for screening of drugs that enhance chemosensitivity by blocking efflux and enhancing drug retention. This article reviews recent developments in our understanding of the multiple drug resistance phenotype and the use of flow cytometry for the study of drug efflux and its modulation in human tumor cells. PMID- 12094477 TI - Reticulocyte analysis by flow cytometry and other techniques. AB - Enumeration of peripheral blood reticulocytes is an essential part of the diagnosis and management of anemic patients, since the number of reticulocytes in the peripheral blood reflects the erythrocytic activity of the bone marrow. Reticulocyte enumeration using flow cytometric methodology is rapidly replacing the inaccurate, imprecise manual counting technique used in the past. This article explores the pathophysiology of the reticulocyte, the various means of counting reticulocytes, and the diverse clinical applications of reticulocyte data. PMID- 12094478 TI - Flow cytometry of platelets for clinical analysis. AB - Platelets are small, non-homogenous cells with distinctive surface features important to their essential role in hemostasis. The surface membrane is dynamic, and changes remarkably in lipid asymmetry and receptor expression on triggering of the activation process. There are also extensive and rapid intracellular changes in platelets as a result of biochemical activation through calcium fluxes, phospholipase activity, kinase activity, and phosphorylation mechanisms that lead to release of storage granule contents and generation of fast-acting prostaglandins, all in a matter of seconds after stimulation with a strong agonist. These characteristics make the platelet an interesting but difficult cell to study, and the explosion of knowledge over the last two decades has been fueled in large part by the application of flow cytometry techniques. Clinical applications of flow cytometry analysis of platelets have been pursued in individual specialized medical centers, but have not found widespread practice in clinical laboratories, mostly because of difficulties in standardization of techniques and the inherent biovariability in comparing normal to abnormal platelets. Despite these hurdles, it seems certain that flow cytometry analysis of platelets in pathological states will continue to evolve into more practical and robust procedures that will eventually become standard hematologic assays rather than specialized research tools. PMID- 12094479 TI - Hematopoietic cellular therapy: implications for the flow cytometry laboratory. AB - Advances in hematopoietic stem cell transplantation, gene therapy, and immunotherapy have necessitated a host of novel monitoring procedures. Cell sorting is also coming of age as a clinical procedure designed to organize hematopoietic grafts for specificity of cellular components designed for individual patients or diseases. This article has focused on these novel developments in their historical context. The next generation of transplantation flow cytometry promises to be an exciting one. PMID- 12094481 TI - [Believing in laparoscopy]. PMID- 12094482 TI - [Mechanisms of ureteral contraction-relaxation in nephritic colic]. AB - OBJECTIVE: From the clinical perspective, pharmacological research on ureteral dynamics--the contraction-relaxation mechanisms of the ureter--, has been prompted by renal colic. Studies of the events triggered by the calculus have been conducted to identify the pharmacological aspects that permit pain relief, passage of the calculus and protection of the kidney from injury caused by acute obstruction. METHODS: Many and varied studies on ureteral pharmacology, both in vivo and in vitro, have been carried out since the beginning of the 20th century. Numerous studies on the anatomical, neurophysiological and hormonal aspects, and different neuroactive substances have attempted to identify the behavior of the ureter under normal and pathological conditions. Since 1967, the Urology Department of the 'Jimenez Diaz' Foundation has investigated the mechanisms of ureteropelvic dynamics utilizing techniques developed at this institution, such as pyelomanometry, pressure/flow to constant pressure studies and antegrade pyelography. RESULTS: The different behavior of the ureter and the stimuli it receives according to the zone analyzed, are known. The classical neurohumoral mechanisms (adrenergic and cholinergic), ureteral pacemaker, effects of urinary pH and infection, action of glucagons, prostaglandins, as well as the new mechanisms of action mediated by substances and non-adrenergic non-cholinergic (NANC) receptors (tachykinins, nitric oxide (NO), vasoactive intestinal peptide (VIP), neuropeptide Y, etc.) have led to new concepts of ureteral contraction relaxation. CONCLUSIONS: The current therapeutic strategy in renal colic utilizes non-steroidal antiinflammatory drugs (NSAID) and extracorporeal shock wave lithotripsy (ESWL) in situ for analgesia and resolution. Recent insight into ureteral dynamics attempt to promote the development of new drugs for enhanced and more effective management of renal colic. PMID- 12094480 TI - Emerging technology and future developments in flow cytometry. AB - The authors' view of the future of flow cytometry is based on a belief that the single most important aspect flow cytometry offers to the investigator is high speed interrogation of correlated measurements on a cell-by-cell basis. Over the next several years, an enormous increase in the capabilities of cytometry in general, and flow cytometry in particular, is likely to occur. A brief description of some of those capabilities is the subject of this article. PMID- 12094483 TI - [Biomolecular markers in renal adenocarcinoma]. AB - OBJECTIVE: To review the literature and analyze the main contributions in molecular markers in renal adenocarcinoma. METHODS: We reviewed the literature over the last 10 years on molecular markers in renal adenocarcinoma (Medline 1991 2001). Outstanding papers published before 1991, included in the references of the selected articles, were also reviewed. RESULTS: A large number of biomolecular markers have been studied in renal adenocarcinoma with results that are often contradictory. Some proteins have been associated with different histopathological parameters and with tumor prognosis, although their clinical importance has yet to be established. CONCLUSIONS: Although the traditional prognostic factors, such as tumor grade and stage, provide important information on the outcome of renal adenocarcinoma, they are insufficient to predict the clinical behavior of this tumor type with absolute certainty. The foregoing has prompted the study of a variety of biomolecular markers that might predict the biological behavior of the tumor and identify the patients at a higher risk for tumor recurrence or death from the tumor. Despite the promising results obtained with biomolecular markers in renal adenocarcinoma, further studies are warranted to determine the prognostic value of these markers. PMID- 12094484 TI - [Complications after radical cystectomy in patients with bladder carcinoma]. AB - OBJECTIVE: To describe the frequency and type of complications after radical cystectomy for cancer of the urinary bladder and their relationship with the different types of urinary diversion techniques utilized. METHODS: We conducted a retrospective study of 166 patients who underwent radical cystectomy in the same department of urology over a period of 12 years. The intraoperative, early, late complications and the need for postoperative treatments are analyzed according to percentages, main trends, scatter and confidence interval. RESULTS: 87.3% of the patients were male. The most common tumor stage was pT3 (37.9%). Transintestinal urinary diversion was performed in 81.1%; the most common being cutaneous ureteroileostomy (44%) and external diversion (64.1%). 16.7% presented hyperchloremic metabolic acidosis. Intraoperative, early and late complications were observed in 53.6%, 47.5% and 71.1% of the survivors for each time period, respectively. 28.3% of the patients required some type of medical treatment in the course of their disease and 30.7% required some type of surgical or percutaneous procedure. The most frequent complications were intraoperative bleeding (50%), respiratory complications early postoperatively (12.6%) and chronic urinary infection as late complication (30.7%). CONCLUSIONS: The high percentage of complications in our series is basically due to the fact that all types of complications, whether directly related or not to the disease and surgical procedure, were included. There was a higher incidence of late complications, the survival time being an important factor that influences the presentation of these complications. Complications are directly related to the type of urinary diversion performed, patient age, tumor stage and previous morbidity. The appearance of complications carries a high percentage of long-term complementary medical treatments or surgical and percutaneous procedures. PMID- 12094485 TI - [Value of gray scale ultrasonography in the early diagnosis of urologic complications of renal transplantation]. AB - OBJECTIVE: To determine the significance of gray scale ultrasound as a tool for diagnosis, follow-up and treatment of urological complications of renal transplantation based on the experience of our institution. METHODS: A retrospective, longitudinal and observational study was carried out. We reviewed the patients' perioperative ultrasound scans and their respective urological complications from January 1, 1982 to January 1, 2000. The patients were consecutively taken from the kidney transplant registry of the Urology and Nephrology and Transplant Departments. We describe the ultrasound findings of normal functioning grafts as well as those with urological complications, such as fluid collections (lymphocele, hematoma, urinoma and abscess), uronephrosis and its possible causes, and symptomatic vesicourethral reflux. RESULTS/CONCLUSIONS: Diagnostic and therapeutic algorithms in kidney transplant patients have been changed since the advent of ultrasound in our country in 1981. Rapid diagnosis and better therapeutic options have been the hallmarks of ultrasound. Added advantages are: it is low-cost, non-invasive, not time consuming. It can be performed regardless of kidney function, can be repeated as many times as required, subsequent scans can be compared. It can be carried out in special care units. The superficial location of the graft makes it highly sensitive. Its disadvantages are low specificity to identify either the nature of the fluid collections or the precise site of urinary tract obstruction, apart from depending on the skill of the operator. Specialists should be familiar with the surgical anatomy of the kidney graft and its variations in order to utilize completely its diagnostic and therapeutic potentials. PMID- 12094486 TI - [Therapeutic advantages of rigid transurethral ureteroscopy in ureteral lithiasic pathology: retrospective study of 735 cases]. AB - OBJECTIVE: To present the results achieved by rigid transurethral ureteroscopy for the management of ureteral calculi over a period of 10 years. METHODS: From January 1991 to November 2000, 735 rigid transurethral ureteroscopy procedures for ureteral calculi were performed in our Lithotripsy Unit. The rigid ureteroscopes utilized ranged from 9.5-11.5 F. There was a higher prevalence of male patients (63%). The mean age was 49.9 years. Calculi were more frequently localized in the pelvic ureter (74.2%). The mean maximum diameter of the calculi was 9.6 mm and caused moderate to severe uropathy in 78.6% of the cases and functional impairment in 3.7% of the cases. After performing ureteroscopy, a double-J catheter was left indwelling in 65.8% of the patients. The patients were discharged from hospital usually 12-18 h after the endoscopic procedure. RESULTS: Of the 735 ureteroscopies performed, satisfactory results were achieved in 676 cases (92%); stone resolution was not achieved in 59 cases. For complete resolution of the calculi, mechanical or electrokinetic fragmentation was used in a high proportion of patients (56.3%). Removal of the stone or stone fragments was mainly by forceps (79.7%); the Dormia basket was used in 18.2% of the cases. The complications occurred during or immediately after the procedure and were minor (10.7%), although there were 3 patients with major complications (2 ureteral eversion and one case of ureteral avulsion). CONCLUSIONS: In our view, rigid transurethral ureteroscopy is a useful, safe and efficient technique in the treatment of calculi in the pelvic ureter, and in selected cases of calculi in the sacral or lumbar level, after failed ESWL. Rigid transurethral ureteroscopy achieves excellent resolution and the complications are scanty. PMID- 12094487 TI - [Initiation of laparoscopic nephrectomy]. AB - OBJECTIVE: This work is reported to encourage experienced urologists to take advantage of a laparoscopic nephrectomy technique that has evolved into a remarkably inexpensive, uncomplicated routine procedure almost comparable with the gallbladder operations of general surgery. METHODS: The patient is placed in the lumbotomy position. The unsleeved, uncuffed hand is introduced through a mini McBurney incision in the iliac fossa. The first 12 mm trocar is introduced just above the posterior part of the iliac crest. The second is placed anteriorly some 10 to 12 cm from the first. The kidney is released by extraperitoneal manual dissection. The artery is clipped and sectioned and then, without dissecting the vein, the rest of the pedicle is sectioned en bloc with the endostapler. However, if the artery is not found, then the whole pedicle is sectioned en bloc. Lastly, the ureter is sectioned. The kidney is then introduced into a simple plastic bag (open urine bag) and digitally fragmented for easy removal through the incision. RESULTS/CONCLUSIONS: The oblique McBurney incision is also useful in the event reconversion to an open lumbotomy is necessary. As we described in 1994, manual manipulation significantly simplifies the procedure. Operators soon gain confidence when they appreciate in practice the surprising versatility of the technique. Because most of the procedure is done under visual control, operator precision soon increases and successive operation times shorten considerably. Operation costs are lower because an expensive inflatable cuff or sleeve is unnecessary, only two trocars are required and a standard urine bag serves to receive the kidney. We are sure that once urologists experienced in conventional surgery try the procedure, they will wonder why they did not adopt it earlier. PMID- 12094488 TI - [Anterograde endopyelotomy]. AB - OBJECTIVE: To analyze the results obtained with antegrade endopyelotomy and compare them with those reported in the literature. METHODS: 16 patients with ureteropelvic junction (UPJ) obstruction treated by antegrade endopyelotomy were evaluated. RESULTS: In our series the success rate was 81.3%. Avulsion of the UPJ was observed in one patient and required conversion into an Anderson-Hynes pyeloplasty. Two cases of re-stenosis were diagnosed within the first three months. CONCLUSIONS: Long-term follow-up shows that antegrade endopyelotomy can be indicated as the treatment of choice for primary and secondary UPJ strictures. PMID- 12094489 TI - [Synchronous and primary adenocarcinoma: renal tumor and gallbladder incidentaloma]. AB - OBJECTIVE: To report a case of two synchronous primary adenocarcinomas involving two different organ systems. METHODS/RESULTS: The urological assessment of a patient with hematuria disclosed an intravesicular polypoid mass and a right renal mass that were treated by surgery. CONCLUSIONS: The histopathological findings demonstrated a clear cell adenocarcinoma of the kidney and another primary adenocarcinoma of the gallbladder. The patient has a prolonged survival. PMID- 12094490 TI - [Obstructive urinary symptoms associated with giant cyst of seminal vesicle]. AB - OBJECTIVE: Seminal vesicle cysts are rare diseases and frequently cause no symptoms. An acquired giant seminal vesicle cyst in a 73-year-old man is presented. Bladder outlet obstruction was the main complaint, mimicking that arising from the prostate. METHODS: Ultrasonography and computed tomography revealed a well-defined, homogeneous, retrovesical cystic mass. Urinary tract was normal. RESULTS: A CT-guided drainage of the cystic mass was performed and urinary symptoms disappeared. The cystic mass recurred at 6 months' follow-up. Treatment was by surgery, which confirmed the seminal vesicle origin of the cyst. CONCLUSIONS: Acquired seminal vesicle cysts may cause bladder outlet obstruction. PMID- 12094491 TI - [Intraureterocele transitional tumor synchronous with contralateral ureteral tumor: obstructive uropathy in panurothelial disease]. AB - OBJECTIVE: To present a case of urothelial tumor inside a simple ureterocele with a coexisting contralateral tumor that caused bilateral obstructive uropathy. METHODS: Follow-up evaluation of a 74-year-old male patient with superficial tumor of the urinary bladder diagnosed 4 years earlier, revealed renal failure and bilateral obstructive uropathy. The complementary tests showed a simple left ureterocele containing a lesion suggestive of a tumor and a right ureteral tumor. RESULTS: After biopsy confirmed coexisting urothelial disease at 3 levels (intraureterocele, contralateral ureter and in situ carcinoma of the bladder), the patient was submitted to cystoprostatectomy, nephroureterectomy and en bloc segmental ureterectomy. CONCLUSIONS: Multicentric synchronous urothelial tumors warrant a close and long follow-up of the entire urinary tract. Among the possible sites of presentation, few cases of tumors inside a ureterocele have been reported. The case described herein, with this uncommon site of presentation, is an example of panurothelial disease. PMID- 12094492 TI - [Fulminant sepsis caused by Clostridium perfringens of urologic origin]. AB - OBJECTIVE: To present a case of fulminant sepsis caused by Clostridium perfringens of urological origin. METHODS: An 81-year-old diabetic female (the only factor of immunodepression) presented complicated renal colic two days later with fulminant and fatal sepsis caused by Clostridium perfringens with signs of disseminated intravascular coagulation. RESULTS: The patient died one hour after the presentation of disseminated intravascular coagulation despite attempts to resuscitate the patient in the emergency department. Due to the fulminant course of the condition, it was not possible to demonstrate the presence of massive intravascular hemolysis characteristic of these conditions. Blood cultures obtained immediately after the patient died were positive for Clostridium perfringens. CONCLUSIONS: Sepsis is a possible complication of infection from Clostridium perfringens. It is more frequent in immune-depressed patients and carries a high mortality despite medical and surgical treatment. Although it is not the most frequent, the genitourinary tract is a known portal of entry that should not be forgotten as in the case described herein. PMID- 12094493 TI - [Glands necrosis caused by necrotizing granulomatous arteritis]. AB - OBJECTIVE: To report a case of necrosis of the glans penis caused by necrotizing granulomatous arteritis, an uncommon form of presentation of this vasculitis that basically affects the upper and lower respiratory tracts and glomerulus. Furthermore, it produces ocular inflammatory lesions and in 50% of the cases skin lesions. The mean age at presentation is around 40 years and affects both males and females in the same proportion. METHODS: A 68-year-old male consulted for pain and change in the color of the penis he had noted two months earlier, and gradually increasing difficulty in voiding. Patient examination revealed a hard necrotic lesion in the distal two thirds of the penis with secondary stenosis of the meatus. We review the literature on the different forms of presentation of this condition and the differential diagnosis of ischemic penile lesions. RESULTS/CONCLUSIONS: In patients with an ischemic lesion of the glans penis, we must consider among other causes iatrogenic lesions, use of constricting devices, gangrenous pyoderma and diseases of small vessels. According to the literature, penile involvement due to necrotizing granulomatous arteritis is uncommon. PMID- 12094494 TI - [Neuroendocrine small cell carcinoma of the bladder. Review of the literature and report of a case]. AB - OBJECTIVE: A case of small cell neuroendocrine carcinoma of the urinary bladder is described. Treatment was by transurethral resection of the bladder tumor in two steps and adjuvant chemotherapy with cisplatin (CDDP) plus etoposide (VP-16) and radiotherapy. To date, 152 cases have been reported. The literature on this type of lesion is reviewed. METHODS/RESULTS: A 54-year-old male was referred to our Service with a complaint of gross hematuria. The radiologic examination showed a lesion on the right aspect of the bladder, close to the bladder neck. Urethroscopy revealed a non-papillary broad-based tumor involving the right lateral wall. TUR of the bladder tumor was performed and an initial diagnosis of small cell neuroendocrine carcinoma was made. Complementary studies showed no widespread disease. The patient refused major open surgery and accepted treatment planning at the preoperative diagnosis of T2-3N0M0 as described above. The patient remains disease-free 28 months after the operation. We also report our experience in a patient free of disease diagnosed at stage pT3BN0M0 and 84 months' follow-up, treated with partial cystectomy and adjuvant M-VAC chemotherapy. CONCLUSIONS: Small cell neuroendocrine carcinoma is an uncommon, usually aggressive neoplasia of the bladder. Patients that are treated have a median survival of 13 months. Cisplatin-based chemotherapy appears to be the only factor associated with a better survival. Relationship between clinical stage and survival has not been demonstrated, which suggests that micrometastatic disease is usually present at presentation in clinically localized tumors and systemic metastases are the major cause of mortality. We believe that partial cystectomy with adjuvant chemotherapy is a valid procedure in favorable cases that allows preservation of the bladder and a high quality of life. PMID- 12094495 TI - [Preputial neurilemoma]. AB - OBJECTIVE: To present a case of preputial neurilemoma. METHODS: A 23-year-old man presented with a small, nodular, preputial mass that he had noted three years earlier. RESULTS: The tumor was excised under local anesthesia without complications. Currently, no evidence of tumor recurrence has been observed. CONCLUSIONS: Preputial nuerilemoma is rare. Diagnosis is based on the findings of the pathological study. PMID- 12094496 TI - [Metabolic syndrome and its complexity]. AB - Changes in human lifestyle over the last century have produced a dramatic increase of diabetes and obesity worldwide. These two chronic diseases are components of the metabolic syndrome, which is associated with high incidence of cardiovascular disease. Genetic and environmental factors such as high calories diet and inactivity are clearly associated with the metabolic syndrome. The science of complex adaptative systems provides important concepts and perspectives to better understand the pathophysiology and the multi-dimensional approach of the treatment of "diabesity". New treatment frameworks are needed to improve the effectiveness of clinicians in promoting behavioural changes and therapeutic adherence. Goal setting should focus on the individual with a metabolic syndrome and not only to the biological and hemodynamic components of the disease. We should take into account the life circumstances of the patient, his motivations and his therapeutic priorities. PMID- 12094497 TI - [Metabolic syndrome: from specialists to the first aid physician]. AB - Numerous algorithms originating from evidence-based medicine already exist; in spite of this, metabolic syndrome management remains problematical and needs to be improved. Specialists, working interactively and referring to clinical scenarios, pooled their specific areas of expertise in a transverse perspective of the metabolic syndrome management by the primary care physician. This concerted multidisciplinary approach has clearly revealed that a bio-psycho social approach is absolutely essential in the management of this type of patients. PMID- 12094498 TI - [Dyslipidemia and metabolic syndrome]. PMID- 12094499 TI - [Obesity, overweight, and diet]. PMID- 12094501 TI - [Treatment compliance, key element in the management of arterial hypertension]. PMID- 12094500 TI - [Follow-up, therapeutic education, and pharmacologic treatments of patients with type 2 diabetes. Various critical views]. AB - Several factors have to be taken into account for successful therapeutic patient education: patients' acceptance of the illness; an interactive educational process, the inclusion of therapeutic objectives in the short, medium and long term which should be planned with the patient. There are numerous facets in the treatment of diabetes, but the common denominator is the dietary programme coupled with daily physical activity. This is the condition sine qua non to be included before the introduction of any pharmacological treatment. The benefits of this latter treatment are: 1) insulin substitution with insulin analogue responding especially quickly, thereby stopping the glycaemic level from falling; 2) insulin secretagogues (Sulfonylurea, Glinides, [Novonorm, Starlix]); 3) agents which raise the sensibility to insulin (glitazones, [Avandia], biguanides); 4) molecules which interfere with the absorption of the glucose at the digestive level (biguanides and alphaglucosidase inhibitors). PMID- 12094503 TI - [Resilience: a concept covering new perspectives on long-term patient care]. PMID- 12094504 TI - [Preparation and follow up of candidates for surgical treatment of obesity: pre- and postoperative courses]. PMID- 12094502 TI - [Paradigm of complexity and chronic disease. The case of metabolic syndrome]. PMID- 12094505 TI - [Sexual life, future of the couple, and contraception after voluntary pregnancy termination. Prospective study in Geneva (Switzerland) with 103 women]. AB - OBJECTIVE: Study the impact of termination of pregnancy on women sexuality, contraceptive behaviour and couple relationship. DESIGN: Prospective qualitative and quantitative study. SUBJECTS: 103 women requiring induced abortion by vacuum aspiration who were interviewed one to three weeks before the operation and again six months later. The interview was performed by a questionnaire including open and closed questions and psychological tests (Locke-Wallace and Horowitz' IES). RESULTS: A majority of women (n = 101) had already used contraception. During the cycle that had resulted in pregnancy, more than a half (n = 58) had actually used contraception and 1/3 had not use any contraception at all. Six months later, the majority (n = 86) used contraception, while only 16 did not. Condom was only rarely used (n = 11). After termination of pregnancy, 14 couples of 84 separated. The majority of women (n = 72) did not report changes in their sexual behaviour. On the other hand, 1/5 of women presented a decrease in sexual desire and orgasm troubles. About 1/3 described psychosomatic symptoms, but a majority (n = 92) was not traumatised. CONCLUSIONS: The great majority of women seems to tolerate well the termination of pregnancy. A minority presents sexual dysfunction, psychosomatic symptoms, which persist six months after intervention. As per contraception, the majority of women does protect herself from pregnancy but a percentage still does not protect itself at all or only in a noneffective or less effective way. After termination of pregnancy, the issue of protection from AIDS and sexuality-transmitted diseases seems to be less important. PMID- 12094506 TI - [Hepatitis B vaccination in adolescents: what do students know and who is their key informant?]. PMID- 12094507 TI - [Sexually transmitted diseases: medico-legal and medico-ethical features]. PMID- 12094508 TI - Alters in dissociative identity disorder. Metaphors or genuine entities? AB - How should the different identities (i.e., alters) that are thought to be typical for dissociative identity disorder (DID) be interpreted? Are they just metaphors for different emotional states or are they truly autonomous entities that are capable of willful action? This issue is important because it has implications for the way in which courts may handle cases that involve DID patients. Referring to studies demonstrating that alters of DID patients differ in their memory performance or physiological profile, some authors have concluded that alters are more than just metaphors. We argue that such line of reasoning is highly problematic. There is little consensus among authors about the degree to which various types of memory information (implicit, explicit, procedural) may leak from one to the other alter. Without such theoretical accord, any given outcome of memory studies on DID may be taken as support for the assumption that alters are in some sense "real." As physiological studies on alter activity often lack proper control conditions, most of them are inconclusive as to the status of alters. To date, neither memory studies nor psychobiological studies have delivered compelling evidence that alters of DID patients exist in a factual sense. As a matter of fact, results of these studies are open to multiple interpretations and in no way refute an interpretation of alters in terms of metaphors for different emotional states. PMID- 12094509 TI - Posttraumatic stress disorder following cancer. A conceptual and empirical review. AB - Life-threatening illness has recently been recognized as a stressor that can precipitate posttraumatic stress disorder (PTSD). This development has raised questions over the extent to which the PTSD diagnosis is applicable to the psychological reaction to being diagnosed with cancer. This paper identifies the core conceptual issues pertaining to cancer-related PTSD, critically reviews the empirical literature on PTSD following cancer, and considers the possible mechanisms and course of PTSD following a diagnosis of cancer. Specific issues that need to be considered in the assessment and treatment of cancer-related PTSD are reviewed. This review highlights that there is a need for stronger empirical base to guide clinical management of PTSD in cancer patients. PMID- 12094510 TI - Treatment of behavior problems in preschoolers: a review of parent training programs. AB - The literature for parent training interventions with conduct-problem preschool aged children is reviewed. Although previous reviews have identified the preschool years as a critical point for intervention [J. Clin. Psychol. 21 (1992) 306; School Psychol. Rev. 22 (1993) 437], there still remains a paucity of well designed outcome studies that have focused exclusively on preschoolers. Interventions that utilize parents as active sources of change remain predominant. The review reveals the need for further research to improve accessibility and delivery of interventions. Treatments that have undergone modification in their delivery format (e.g., use of telephone and videotape) were also identified and critically evaluated. Results of the review indicate that many studies are still accompanied by methodological limitations. Although promising interventions exist for young conduct-problem children, it is recommended that treatment effectiveness for preschoolers be evaluated more rigorously. PMID- 12094511 TI - Survivors of childhood brain tumors: behavioral, emotional, and social adjustment. AB - This paper reviews the literature on the psychological adjustment and quality of life in children who survive brain tumors. A total of 31 studies were reviewed. Findings are discussed in terms of the rates of general psychological adjustment, internalizing behavior problems, externalizing behavior problems, social competence, correlates or predictors of adjustment, and quality of life among survivors. Although these survivors appear to be at risk for compromised social competence and long-term quality of life, reports in the literature on rates of psychological adjustment in this population vary widely. Limitations in the current literature are discussed including inadequate assessment techniques, lack of appropriate comparison groups, and small sample sizes. Directions for future research are offered. PMID- 12094512 TI - Memory biases in the anxiety disorders: current status. AB - Information-processing models of emotional disorders suggest that anxious individuals may be characterized by a memory bias for threat-relevant information. This paper reviews and synthesizes evidence for explicit (conscious) and implicit (unconscious) memory biases in the anxiety disorders. Our review suggests variations among the anxiety disorders for explicit memory biases. Specifically, there is support for explicit memory biases for threat-relevant information in panic disorder (PD), particularly when information has been deeply encoded, but not in social phobia (SP) or generalized anxiety disorder (GAD). The few available studies suggest the presence of explicit memory biases in posttraumatic stress disorder (PTSD) and obsessive-compulsive disorder (OCD), but further research is needed. In contrast, some degree of support for implicit memory biases has been demonstrated for each of the anxiety disorders. Inconsistencies in the existing literature, topics worthy of future research attention, and directions for revising existing information-processing models of anxiety are discussed. PMID- 12094513 TI - Opening doors. PMID- 12094514 TI - Managing delirium in the palliative care of older people. PMID- 12094515 TI - Growing old with dignity: a concept analysis. PMID- 12094516 TI - Epilepsy and services for older people. AB - Although epilepsy can occur at any age, almost a quarter of people affected are over 60 years old. It is important for nurses working with older people to understand the full implications of a diagnosis of epilepsy, which can have a devastating effect on quality of life. PMID- 12094517 TI - Assessing the need for restraint in older people. PMID- 12094518 TI - [Prediction of in-stent restenosis by measurement of fractional flow reserve and the influence of diabetes mellitus]. AB - OBJECTIVES: This study tried to predict in-stent restenosis by the measurement of fractional flow reserve and to evaluate the effect of diabetes mellitus control on the relationship between in-stent restenosis and fractional flow reserve. METHODS: Fractional flow reserve was measured in 62 patients (mean age 61 +/- 9 years, 32 with angina pectoris and 30 with myocardial infarction) after stent implantation. The patients were divided into three groups according to the value of HbA1c and fasting blood sugar (FBS): Group P (HbA1c > or = 6.5% or FBS > or = 126 mg/dl, n = 15), Group G (6.5% > HbA1c > or = 5.8% or 126 > FBS > or = 100 mg/dl, n = 12), and Group N (HbA1c < 5.8% and FBS < 100 mg/dl, n = 35). The relationship between fractional flow reserve at the end point of stent implantation and target lesion revascularization (TLR) was evaluated. RESULTS: In all patients, fractional flow reserve was 0.89 +/- 0.17 and percentage diameter stenosis was 17.6 +/- 11.6% after stent implantation. The TLR rate was 21%. Fractional flow reserve was significantly lower in patients with TLR (0.85 +/- 0.08 vs 0.91 +/- 0.06, p < 0.01). Fractional flow reserve was significantly lower in patients with TLR in Groups N and G (Group N: 0.85 +/- 0.04 vs 0.91 +/- 0.06, p < 0.05, Group G: 0.77 +/- 0.06 vs 0.91 +/- 0.05, p < 0.01). Fractional flow reserve showed no significant difference between patients with TLR and without TLR in Group P (0.85 +/- 0.10 vs 0.87 +/- 0.08, p = NS). In patients with optimal results (fractional flow reserve > or = 0.90), TLR rate was 60% in Group P, 0% in Group G and 0% in Group N. CONCLUSIONS: Fractional flow reserve after stent implantation is useful for the prediction of TLR in patients without diabetes mellitus. However, diabetic control in patients with diabetes mellitus might be attributable to TLR rather than fractional flow reserve. PMID- 12094519 TI - [Preischemic infusion of alpha-human atrial natriuretic peptide elicits myoprotection through a nitric oxide-dependent mechanism]. AB - OBJECTIVES: Alpha-human atrial natriuretic peptide (alpha-hANP) has been used to treat patients with heart failure due to its natriuretic and vasodilatory activities. Recent reports have suggested that alpha-hANP generates nitric oxide (NO) that is known to be involved in myoprotective mechanisms. In this study, the effects of preischemic infusion of alpha-hANP against reperfusion injury were evaluated. METHODS: Isolated rat (Sprague-Dawley rat, age 8-10 weeks, weight 260 340 g) hearts were subjected to Langendorff perfusion with buffered Krebs Henseleit solution and were divided into three groups: Six hearts were treated with 0.1 microM of alpha-hANP for 10 min (Group H), six hearts with 1 mM of a NO synthetase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) for 5 min before alpha-hANP (Group L), and six hearts served as the controls with no interventions (Group C). All groups were then subjected to 20 min of global ischemia followed by 120 min of reperfusion. Left ventricular pressures and coronary flow were measured throughout the experiment and infarct size was evaluated at the end of the experiments. RESULTS: Treatment with alpha-hANP significantly reduced infarct size as compared to control hearts whereas pretreatment with L-NAME almost reversed the preventive effect (Group C = 42.7 +/- 2.3%, Group H = 26.1 +/- 2.8% *, Group L = 39.0 +/- 1.6%; * p < 0.01 vs Group C). There were no significant differences in left ventricular pressure and coronary flow between the three groups. CONCLUSIONS: Preischemic infusion of alpha-hANP may provide myoprotective effects against postischemic reperfusion, possibly through a NO-dependent mechanism. PMID- 12094521 TI - [Quadricuspid pulmonary valve with valvular stenosis and regurgitation identified by transthoracic echocardiography: a case report]. AB - A 74-year-old man was admitted to our hospital complaining of dyspnea. Parasternal transthoracic echocardiography showed a quadricuspid pulmonary valve above the aortic valve and a hypoplastic accessory cusp between the right and left cusps of the pulmonary valve. The pulmonary valve ring diameter was normal (26 mm) but the transvalvular peak velocity was 3.5 m/sec, suggesting a pressure gradient of 49 mmHg across the pulmonary valve. The pulmonary valve had thickening and decrease in mobility of the leaflets without complete closure during diastole, and severe pulmonary regurgitation was present. Heart failure was treated successfully with digitalis and diuretics. Quadricuspid pulmonary valve is difficult to identify using transthoracic echocardiography because of the anatomical features. In this case, the dilated main pulmonary artery caused the pulmonary valve orifice to shift anteriorly, allowing visualization of the short-axis view of the pulmonary valve. PMID- 12094520 TI - [Evaluation of coronary flow reserve using Doppler guide wire in patients with ampulla cardiomyopathy: three case reports]. AB - Coronary flow velocity recordings were obtained in three patients with ampulla (Takotsubo) cardiomyopathy using Doppler guide wire. Immediately after admission, coronary angiography demonstrated no significant stenosis and normal coronary flow in the epicardial coronary artery. Left ventriculography showed abnormal wall motion of the left ventricle mimicking ampulla shape in all patients. The coronary flow velocity pattern was normal, but coronary flow reserve of the left anterior descending coronary artery was decreased by adenosine administration in all patients. The coronary flow reserve in the left circumflex and right coronary artery also decreased. Cardiac catheterization was performed during the follow-up period in two patients. Left ventriculography revealed almost normal wall motion of the left ventricle. The coronary flow reserve normalized in one patient. PMID- 12094523 TI - [A 37-year-old man with giant negative T waves and increased serum creatine kinase]. PMID- 12094522 TI - Primary cardiac sarcoma mimicking benign myxoma: a case report. AB - A 53-year-old woman presented with a primary cardiac sarcoma mimicking benign myxoma manifesting as progressive heart failure. Transesophageal echocardiography disclosed two separate tumors in the left atrium which appeared just like myxoma, except for the origin and multiple growth profile. Three separate tumors were identified during the operation arising from the posterior wall of the left atrium, mitral valve orifice and left atrial free wall. The histological diagnosis was malignant undifferentiated sarcoma. Six months later, she noticed dyspnea and arm numbness due to local recurrence of cardiac tumor and brain metastasis. She died suddenly 6 months after the surgery during admission. The clinical and echocardiographic findings are crucial to discriminate malignant cardiac tumors from benign myxoma. Rapidly progressive clinical course, multiple tumor growth and non-septal attachment of the tumor all suggest a malignant profile of the primary cardiac tumors. Careful and precise preoperative evaluation, including echocardiographic survey of the tumor origin and multiple growth profile, are essential to identify malignant cardiac tumor. PMID- 12094524 TI - [A 68-year-old man with acute aortic regurgitation and left-sided heart failure]. PMID- 12094525 TI - The volunteers: the first human biopsy studies of TCDD from Agent Orange exposure. PMID- 12094526 TI - Vietnam and Agent Orange revisited. PMID- 12094527 TI - On the plausible association between environmental conditions and human eye damage. AB - The increase in solar ultraviolet radiation can have various direct and indirect effects on human health, like the incidence of ocular damage. Data of eye damage in residents of three suburban regions in Greece and in two groups of monks/nuns and fishermen are examined here. The statistics performed on these data provides new information about the plausible association between increased levels of solar ultraviolet radiation, air-pollution at ground level, and the development of ocular defects. PMID- 12094528 TI - Detection of indoor PCB contamination by thermal desorption of dust. A rapid screening method? AB - Although PCB in caulking materials has been forbidden for many years in most of Europe, including Denmark, there has been continued interest to measure PCB levels in the air of contaminated buildings and blood of the occupants (Mengon and Schlatter 1993, Fromme et al. 1996, Ewers et al. 1998, Currado and Harrad 1998, Gabrio et al. 2000). The relatively low priority for investigations of this contamination is probably due to the small quantities inhaled compared to exposure via food, and the rapid metabolism of the most volatile congeners demonstrated by low concentrations of all congeners in the blood of exposed persons (Ewers et al. 1998, Gabrio et al. 2000). There is, however, evidence that PCB containing caulking materials have been used even during the '90s (Fromme et al. 1996). In Denmark, it is estimated that 75 t PCB is still in buildings (Organization of Sealant Branch's Manufacturers and Distributors 2000). During an investigation of dust from buildings with excessive microbial growth (including 35 rooms from 9 buildings), the analysis of semivolatile compounds by thermal desorption-GC/MS of samples from a single building surprisingly revealed large amounts of PCBs containing 3, 4 and 5 chlorine atoms, 10-20 times the amounts found in samples from other buildings. Extraction of the dust by SFE followed by GC/ECD analysis for 12 PCB congeners showed that there was approximately 20 times the total PCB concentrations in dust from the polluted building compared to the levels in the other buildings. Subsequent headspace analysis of caulking material from the polluted building revealed this to be the source. Shelf dust functions as a passive sampling medium and, thus, can be used as a screening method to detect PCB and other semivolatile pollution indoors. PMID- 12094529 TI - BTX concentrations near a stage II implemented petrol station. AB - A combined monitoring and dispersion modelling methodology was applied for assessing air quality at three different levels of proximity to the selected service station: (I) next to the fuel pumps, (II) in the surrounding environment, and (III) in the background. Continuous monitoring and passive sampling were used for achieving high temporal and spatial resolution, respectively. A Gaussian dispersion model (CALINE4) was used for assessing the road traffic contribution to the local concentrations under different meteorological conditions. It was established that Stage 2 vapour recovery reduces BTX concentrations not only near the pumps, but also in their surrounding environment. However, there is evidence that the efficiency of the system is wind speed dependent. The modelling simulation of the worst case wind scenario revealed the significance of local traffic emissions. It was shown that the traffic contribution even from a single road in the vicinity of the station can, under certain conditions, be higher than the contribution of the station itself to the local BTX levels. Finally, after comparison with previous studies, the concentrations measured near the service station (which was situated in a rural environment) appear to be lower than those observed in busy street canyons in city centres. It can be concluded, although Stage 2 recovery system effectively reduces working VOC losses in service stations, that it will only have a limited positive impact on local air quality if the service station is located in a heavily polluted area. PMID- 12094530 TI - The distribution of the chlorinated solvents dichloromethane, perchloroethylene, and trichloroethylene in the global atmosphere. AB - Dichloromethane, perchloroethylene, and trichloroethylene are commercially important chlorinated solvents whose health and environmental impacts are under scrutiny in the industrial world. Their distributions in the global atmosphere have been computed based on data from the Reactive Chlorine Emissions Inventory (RCEI) project using the Global Balance Environment (GLOBE) model, a 3-D radiative-dynamical-chemical model. Their atmospheric lifetimes, scaled to an observed methyl chloroform lifetime of 4.8 years, are 158 days, 105 days, and 4.3 days, respectively. They have strong interhemispheric gradients, with maximum zonal mean surface concentrations in the winter mid-latitude northern hemisphere of approximately 40 ppt, 9 ppt, and 2.5 ppt, respectively. Their spatial distributions show significant seasonal variability, and are sensitive to vertical mixing by cumulus convection and horizontal mixing by synoptic-scale turbulence. While the model interhemispheric exchange time (1.0 years) and computed atmospheric lifetimes are very sensitive to sub-grid scale diffusion, interhemispheric gradients of the chlorinated solvents are not. The simulated results suggest a greater importance for oceanic emissions of perchloroethylene and trichloroethylene than has previously been assumed. PMID- 12094531 TI - Stable chlorine intramolecular kinetic isotope effects from the abiotic dehydrochlorination of DDT. AB - INTENTION, GOAL, SCOPE, BACKGROUND: Identifying different sources and following reaction pathways of chlorinated organic contaminants in the environment can be challenging, especially when only their concentrations are available. Compound specific stable chlorine measurements of some contaminants have recently been shown to provide additional information and an increased understanding of their biogeochemistry. These studies, however, have been generally limited to volatile molecules. OBJECTIVE: Here, the stable chlorine isotope ratios of the semi volatile pesticide, 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) were investigated. Specifically, the intramolecular stable chlorine isotopic compositions of DDT and the kinetic isotope effect (KIE) for the abiotic dehydrochlorination of DDT to 2,2-bis(p-chlorophenyl)-1,1-dichloroethene (DDE) were determined. METHODS: Selective chemical oxidation of DDT to dichlorobenzophenone (DCBP) and analysis of each compound was used to calculate the stable chlorine isotope ratios of the alkyl and aromatic chlorines in DDT. To determine the KIE for dehydrochlorination, DDT was reacted in a basic solution to yield DDE at 52 degrees C, 60 degrees C, and 72 degrees C for 3, 5, and 5 days, respectively. RESULTS AND DISCUSSION: Significant intramolecular stable chlorine isotopic differences were observed in one sample of DDT where the alkyl and aromatic delta 37Cl values were -5.76 +/- 0.45 and -2.21 +/- 0.24%@1000, respectively. Dehydrochlorination of DDT to DDE in basic solutions at 52, 60, and 70 degrees C resulted in a substantial intramolecular KIE where the alkyl chlorines of DDE shifted by approximately 3%@1000 relative to the alkyl chlorines in DDT. However, no temperature dependence was observed. The KIE, calculated by an iterative program, was 1.009. CONCLUSIONS: Intramolecular differences in the stable chlorine isotope ratios were observed in DDT and this is the first such finding. Dehydrochlorination of DDT yields a measurable and distinct intramolecular stable chlorine KIE. RECOMMENDATION AND OUTLOOK: The results of this study demonstrate the existence of significant intramolecular differences in chlorinated organic compounds. Many other chlorinated semi-volatile and volatile organic contaminants are synthesized from multiple sources of chlorine, and we recommend that similar studies be performed on many such molecules in order to attain a clear understanding of their intramolecular chlorine isotopic differences. The existence of a measurable KIE for the dehydrochlorination of DDT to DDE shows the potential strength of using isotopic measurements to investigate the biogeochemistry of these important compounds. For example, the isotopically depleted aqueous chloride produced by dehydrochlorination of DDT to DDE may be a useful tracer of these reactions in freshwater environments. PMID- 12094532 TI - Perchlorate as an environmental contaminant. AB - Perchlorate anion (ClO4-) has been found in drinking water supplies throughout the southwestern United States. It is primarily associated with releases of ammonium perchlorate by defense contractors, military operations, and aerospace programs. Ammonium perchlorate is used as a solid oxidant in missile and rocket propulsion systems. Traces of perchlorate are found in Chile saltpeter, but the use of such fertilizer has not been associated with large scale contamination. Although it is a strong oxidant, perchlorate anion is very persistent in the environment due to the high activation energy associated with its reduction. At high enough concentrations, perchlorate can affect thyroid gland functions, where it is mistakenly taken up in place of iodide. A safe daily exposure has not yet been set, but is expected to be released in 2002. Perchlorate is measured in environmental samples primarily by ion chromatography. It can be removed by anion exchange or membrane filtration. It is destroyed by some biological and chemical processes. The environmental occurrence, toxicity, analytical chemistry, and remediative approaches are discussed. PMID- 12094533 TI - European scenarios for EUSES regional distribution model. AB - The regional multimedia distribution model incorporated into EUSES 1.0 is used for the estimation of regionally predicted environmental concentrations in different European scenarios: a scenario representing a typical region in the north of Europe (high fraction connected to sewer systems, lower environmental temperature, high fractions of surface water and natural soil and a low fraction for agricultural soil) and another scenario representing a typical region in the south of Europe (low fraction connected to sewer systems, higher environmental temperature, low fractions for surface water and natural soil, and a high fraction for agricultural soil). The two scenarios are based on average data of countries in Northern and Southern Europe, but are not realistic for any specific country located in these regions. Scenario calculations were carried out using these two scenarios in addition to the generic standard region, given in EUSES 1.0 as a default scenario, and the North-Rhine Westphalian region. The substance properties, including emissions, were left unchanged for all scenarios. For a number of substances, the calculated concentrations in both the North and the South of Europe turned out to be higher than those calculated with the standard generic scenario. Thus, the standard scenario cannot be considered as a 'worst case' scenario per se. Uncertainties due to the regional heterogeneity within Europe are high. It is recommended to use these two additional scenarios for an improved estimation of possible concentration ranges in Europe. PMID- 12094535 TI - Towards sets of hazardous waste indicators. Essential tools for modern industrial management. AB - Decision-makers require useful tools, such as indicators, to help them make environmentally sound decisions leading to effective management of hazardous wastes. Four hazardous waste indicators are being tested for such a purpose by several countries within the Sustainable Development Indicator Programme of the United Nations Commission for Sustainable Development. However, these indicators only address the 'down-stream' end-of-pipe industrial situation. More creative thinking is clearly needed to develop a wider range of indicators that not only reflects all aspects of industrial production that generates hazardous waste but considers socio-economic implications of the waste as well. Sets of useful and innovative indicators are proposed that could be applied to the emerging paradigm shift away from conventional end-of-pipe management actions and towards preventive strategies that are being increasingly adopted by industry often in association with local and national governments. A methodological and conceptual framework for the development of a core-set of hazardous waste indicators has been developed. Some of the indicator sets outlined quantify preventive waste management strategies (including indicators for cleaner production, hazardous waste reduction/minimization and life cycle analysis), whilst other sets address proactive strategies (including changes in production and consumption patterns, eco-efficiency, eco-intensity and resource productivity). Indicators for quantifying transport of hazardous wastes are also described. It was concluded that a number of the indicators proposed could now be usefully implemented as management tools using existing industrial and economic data. As cleaner production technologies and waste minimization approaches are more widely deployed, and industry integrates environmental concerns at all levels of decision-making, it is expected that the necessary data for construction of the remaining indicators will soon become available. PMID- 12094534 TI - EuroBionet: a pan-European biomonitoring network for urban air quality assessment. AB - EuroBionet, the 'European Network for the Assessment of Air Quality by the Use of Bioindicator Plants', is an EU-funded cooperative project currently consisting of public authorities and scientific institutes from 12 cities in 8 countries. In 2000, the bioindicator plants tobacco (Nicotiana tabacum Bel W3), poplar (Populus nigra 'Brandaris'), spiderwort (Tradescantia sp. clone 4430), Italian rye grass (Lolium multiflorum italicum) and curly kale (Brassica oleracea acephala) were exposed to ambient air at 90 monitoring sites according to standardised methods. Visible injuries and growth parameters were assessed and the accumulation of toxic substances in leaves determined. The exposure of tobacco resulted in a gradient with low levels of ozone-induced foliar injury in N and NW Europe, and medium to high values in the southern and central regions. The results of heavy metal and sulphur analyses in rye grass samples generally showed low to very low sulphur and low to medium heavy metal concentrations in leaves. In some cities, however, local hot spots of heavy metal contamination were detected. Analyses of the PAH contents in curly kale leaves gave low to medium values, with locally elevated levels at traffic-exposed sites. PMID- 12094536 TI - Environmental knowledge and awareness in secondary Polish education. PMID- 12094537 TI - Contribution of biotechnology to chemical-weapons destruction. Report on the Fourth International Workshop 'Contribution of Biotechnology to Chemical-Weapons Destruction' (Saratov, Russia, September 6-7, 2001). PMID- 12094538 TI - Intensity modulated radiotherapy (IMRT) decreases treatment-related morbidity and potentially enhances tumor control. AB - Intensity modulated radiation therapy (IMRT), a new form of three-dimensional conformal radiation therapy (3DCRT), optimizes the concept of computer-controlled radiation deposition in tumor (target) while sparing adjacent normal structures. A retrospective review was done on the initial 185 patients with tumors in different sites including prostate cancer, head and neck cancer, pediatric tumors, adult brain tumors, and previously irradiated recurrent tumors treated with IMRT. Preliminary findings indicate that IMRT is a new clinically feasible tool in radiation oncology. Treatment-related morbidity profile was favorable. Tumor response, local control, and the ability to palliate previously irradiated patients are encouraging. Intensity modulated radiation therapy will allow dose escalation, leading to better tumor control. PMID- 12094539 TI - Breast cancer in HIV-positive women: a report of four cases and review of the literature. AB - Only 16 cases of breast carcinoma in human immunodeficiency virus (HIV) seropositive patients have been reported in the literature. We report four cases of breast cancer in women seropositive for the HIV and review the literature. Breast cancer is not an acquired immunodeficiency syndrome (AIDS)-defining disease and its incidence is not increased in HIV patients. Most patients reported had a CD4 count, which was above the threshold considered critical for significant immunosuppression suggesting that, HIV related immunocompromise does not have a direct tumorigenic role. Alternatively, the biology of breast cancer appears to be more aggressive in HIV-positive women suggesting a permissive role of the virus in the progression of the cancer. The aggressiveness of the breast carcinoma in HIV-positive women justifies every effort to preserve the dose intensity of treatment in those patients, especially in the current era of more effective HIV treatment which leads to improved survival. PMID- 12094540 TI - A case-control study on risk of changing food consumption for colorectal cancer. AB - OBJECTIVES: To investigate the relative risk factor of food items for colorectal cancer in four time periods through a case-control study in a Chinese rural area. METHODS: Colorectal cancer patients diagnosed at a county cancer center, Hebei Province, China, and non-cancer outpatients with similar age, sex, and place of residence were selected for cases and controls, respectively. There were 102 (93.6%) colorectal cancer patients and 99 (90.8%) outpatients being the cases and controls, respectively in the present investigation, who agreed to be interviewed about their food intake, during a 20-year period, through a food frequency questionnaire. The risks of intake of different food items and lifestyle for colorectal cancer were compared between cases and controls. RESULTS: During the 20-year period, diets of both cases and controls changed with increase in intake of animal foods and fruits, and alcohol consumption tended to increase. In the food items, milk intake showed a protective effect in both males and females, and the odds ratios were 0.38 (95% CI 0.16-0.90) and 0.28 (95% CI 0.10-0.81) for males and females, respectively. A reduced risk of fruit intake could be seen in males, while a reduced risk of vegetables could be observed in females. Meat intake and saturated fats were the prominent risk factors for colorectal cancer in males and females, respectively. A comparison of life habits, showed that tea drinking had a consistent protective effect in females, and the odds ratios were 0.21 (0.08-0.58), 0.23 (0.08-0.67), 0.25 (0.10-0.64), and 0.11 (0.04-0.30) for periods of 20-, 10-, 5-years ago, and current time, respectively. CONCLUSIONS: These findings indicate that change in food consumption is strongly associated with a change in risk of colorectal cancer, and dietary meat has increased the risk of colorectal cancer. Increase in the consumption of milk and fruits may be a significant measure for colorectal cancer prevention in low-incidence areas. PMID- 12094541 TI - Phase II study of docetaxel in patients with pancreatic cancer previously untreated with cytotoxic chemotherapy. AB - In this study, we estimated the response rate, duration of response, and type, severity and reversibility of toxicities in patients with Stage IV adenocarcinoma of the pancreas treated with docetaxel. Twenty-one patients with locally advanced or metastatic pancreatic cancer, previously untreated or treated with surgery or radiation alone, were treated with 100 mg/m2 docetaxel as a 1 hr infusion once every 21 days. All the patients were pretreated with dexamethasone and diphenhydramine. Twenty patients were assessable for both response and toxicity. One patient was assessable for toxicity alone. However, all the patients were assessed for survival. The major side effect of the drug was neutropenia, which required a dose reduction to 75 mg/m2 in approximately half of the patients. Nine patients were hospitalized with neutropenic fever. Fluid retention was not a significant problem. One patient had a partial response lasting for 21 weeks and 7 patients had stable disease. The remaining patients had progressive disease. The median survival for all the patients was 5.9 months. Docetaxel as a single agent showed limited activity against adenocarcinoma of the pancreas. Since the completion of this study, molecular predictors of in vitro response to docetaxel have been described. Confirmation of the clinical relevance of such predictors in humans could allow for the identification of a subgroup of patients with a higher rate of response to docetaxel. PMID- 12094542 TI - Phase I trial of edatrexate in advanced breast and other cancers. AB - Edatrexate (EDX) (10-ethyl-10-deazaaminopterin or 10-EDAM) is an analogue of methotrexate with improved pre-clinical antitumor activity, more selective cellular uptake, and with more extensive formation of intracellular polyglutamate metabolites. A phase I trial in advanced cancer using EDX was initiated to determine the toxicities associated with the use of a biweekly schedule of intravenous EDX, and to carry out a dose escalation to define the maximum tolerated dose employing this schedule. Thirty-four patients were enrolled in this phase I trial. Thirty-three patients were treated in cohorts of at least three patients (except at one dose level, 210 mg m-2, where there were only two patients). Dose escalations of EDX were administered starting with 100 mg m-2, and progressing through 120, 140, 160, 180, 210, 240, and 270 mg m-2. Edatrexate was administered by intravenous infusion over 20 min, and cryotherapy using ice chips was given prophylactically for 5 min before, during, and 15 min after each EDX treatment. The dose-limiting toxicity could not be reached in this study because it had to be closed on account of competing protocols using EDX in combination regimens. Of note though, was that the delivered dose intensity at the 160 mg m-2 week-1, was higher than the previously used or recommended phase II doses. Anemia was mild and white blood corpuscle toxicity was mostly of grade 1 or 2. One patient had grade 4 neutropenia and one had grade 3 thrombocytopenia. Of the non-hematological toxicities, nausea, vomiting, and diarrhea were mild and tolerable. Mucositis, which was the dose-limiting toxicity in previous studies, was seen in 30% of the patients, but was predominantly a grade 1 toxicity. This could have been due to either the different schedule of EDX used in this study or the use of cryotherapy. Substantial antitumor effects were noted, with two near complete complete responses at the 120 and 160 mg m-2 levels. Additionally, six partial responses and one minor response were observed, and stable disease was observed in seven patients. Despite achieving antitumor activity at different dose levels, a clear-cut dose response was not evident at the levels tested. We feel that the biweekly EDX schedule is a tolerable regimen, which allows a higher dose intensity than weekly administration, and that EDX is an active agent for the treatment of patients with metastatic cancer. PMID- 12094544 TI - The effect of nonsteroidal antiinflammatory drugs ibuprofen, flurbiprofen, and diclofenac on in vitro and in vivo growth of mouse fibrosarcoma. AB - For suppression of primary G:5:113 fibrosarcoma growth, three structurally different cyclooxygenase (COX) inhibitors (ibuprofen, flurbiprofen, and diclofenac) were administered intraperitoneally (i.p.) in two regimens starting on day 5 after tumor-cell inoculation. Repeated application of 0.15 mg/mouse/day during 14 consecutive days significantly suppressed the tumor growth and increased the percentage of surviving mice. Similar tendency, however without significant differences, was observed when animals were given 0.5 mg/day for five consecutive days. These results suggest that a time schedule of drug application is important for the therapeutic effect. Suppressive effect of diclofenac and flurbiprofen on tumor growth was also observed under in vitro conditions. We conclude that suppressive effect of these drugs on tumor growth in vivo comprises both direct effects of COX inhibitors on fibrosarcoma cells and indirect effects that are presumably mediated by extratumoral sources. Our findings encourage the use of COX inhibitors in the therapy of fibrosarcoma. PMID- 12094543 TI - Evaluation of cisplatin in combination with a biologic response modifier in a murine mammary carcinoma model. AB - The purpose of this study was to determine the efficacy of intracavitary cisplatin against local regrowth and metastasis after resection of a murine mammary carcinoma and the ability of a biologic response modifier (Virulizin) to enhance chemotherapy. C3H-HeJ mice were injected with Gollin-B tumor cells. Once growth reached 8 mm, tumors underwent marginal resection and the mice were assigned randomly to intraperitoneal (i.p.) cisplatin, Virulizin, a controlled release cisplatin-impregnated sponge (OPLA-Pt), a combination of treatments or no treatment and were evaluated for local regrowth, metastasis, and toxicity at 14 or 60 days after surgery. A possible beneficial interaction was seen between OPLA Pt and Virulizin at 14 days. All cisplatin groups had significant advantages over controls in all variables measured with OPLA-Pt displaying significant advantages over i.p. cisplatin in local recurrence rate, tumor score, survival time, and delay in regrowth at 60 days. No toxicity related to either cisplatin or Virulizin was observed. PMID- 12094545 TI - Lack of cell-cycle specific effects of tumor necrosis factor-alpha on tumor cells in vitro: implications for combination tumor therapy with doxorubicin. AB - Addition of tumor necrosis factor-alpha (TNF-alpha) to chemotherapy enhances tumor response in several treatment modalities. However, it has been shown that TNF-alpha, and several other cytokines, exert inhibitory effects on cell-cycle progression and by doing so may attenuate sensitivity of these cells to cell cycle dependent cytotoxic drugs (e.g., doxorubicin). Here, we determined the cytotoxic effect of TNF-alpha on several tumor cell lines in vitro in combination with doxorubicin (cell-cycle dependent) or melphalan (cell-cycle independent), and its effect on cell-cycle progression. The rat cell lines were prepared from tumors, which were used previously in animal studies, in which synergy was shown between TNF-alpha and the cytotoxic drugs. Results demonstrate that the addition of TNF-alpha to doxorubicin or melphalan in vitro had no attenuating effect on the cytotoxic drugs. Depending on the cell type used, addition of TNF-alpha induced no or only an additive cytotoxic effect. Only the tested rat osteosarcoma tumor cells demonstrated a cell arrest in the G2 phase, which did not result in attenuation of the cytotoxicity of doxorubicin towards these cells. PMID- 12094546 TI - Altered subcellular localization of p53 in estrogen-dependent and estrogen independent breast cancer cells. AB - LCC2, an estradiol-independent tamoxifen (Tax)-resistant subline of MCF-7 human breast cancer cell line, is resistant relatively towards Tax and methotrexate (Mtx). The purpose of the present study is to evaluate the role of p53 in determining this resistance. While MCF-7 is sensitive to and undergoes apoptosis, as determined by propidium iodide stain, by Tax and Mtx, LCC2 is resistant to apoptosis induction by these agents. Both cell lines undergo apoptosis and are sensitive equally to doxorubicin (Adr). p53 cDNA of both sublines was evaluated by polymerase chain reaction (PCR) amplification and sequencing and was found to be of wild-type. p53 mRNA, as well as protein, are elevated markedly in LCC2 as compared to MCF-7 cells. p53 expression was increased by estradiol and Adr, not changed by Mtx, and decreased by Tax and estradiol-deprivation in both sublines. p53 modulation by the various agents, in both sublines, was evaluated by cytochemical staining and subcellular fractionation. This analysis showed that p53 is localized mainly in the nuclear fraction in MCF-7 cells, and in the cytoplasmatic fraction in LCC2 cells. Doxorubicin induced apoptosis in MCF-7 cells along with increase in its nuclear fraction. In contrast, LCC2 underwent apoptosis by Adr despite its cytoplasmatic sequestration. These experiments demonstrate that p53 is sequestered to cytoplasm in the estrogen-independent, Tax resistant LCC2 cells. However, the differences in apoptotic rate between MCF-7 and LCC2 cells do not seem to be dependent on p53. The LCC2 cell line may serve as a useful model for the study of the mechanism of cytoplasmatic sequestration of wild type (wt) p53, its physiologic consequences, and its relation to estrogen independence or Tax resistance of breast cancer cells. PMID- 12094547 TI - Usefulness of technetium-99m tetrofosmin scintimammography to detect breast cancer in mammographically dense breasts. AB - The purpose of this study was to assess the usefulness of techentium-99m tetrofosmin (Tc-TF) scintimammography to detect breast cancer in 32 female Taiwanese patients with indeterminate mammograpic probability of malignancy because of mammographyically dense breasts. All breast masses were removed and final histopathological diagnoses were obtained in all cases. The results showed that Tc-TF scintimammography findings were true-positive in 20 cases, false positive in 1 case, true-negative in seven cases, and false-negative in four cases. The diagnostic sensitivity, specificity, and accuracy were 83, 88, and 84%, respectively, for detecting breast cancer in mammographically dense breasts. In conclusion, Tc-TF scintimammography is a useful tool for detecting breast cancer in patients with indeterminate mammograms because of mammographically dense breasts. PMID- 12094548 TI - Current diagnosis and management of cervical cancer. PMID- 12094549 TI - The proliferation of Drosophila in cancer research: a system for the functional characterization of tumor suppressors and oncogenes. PMID- 12094550 TI - Mitochondrial DNA alterations in cancer. AB - A number of studies have demonstrated the presence of mitochondrial DNA (mtDNA) mutations in cancer cells. In this article, we review mitochondrial genomic aberrations reported in solid tumors of the breast, colon, stomach, liver, kidney, bladder, head/neck, and lung. The tantalizing association of tumors with mtDNA mutations implicates these mutations in the process of carcinogenesis. Alterations in expression of mtDNA transcripts in a variety of cancer types are also reviewed. In solid tumors, elevated expression of mtDNA-genes coding for subunits of the mitochondrial electron respiratory chain may reflect mitochondrial adaptation to perturbations in cellular energy requirements. The role of mtDNA mutations and altered expression of mitochondrial genes in carcinogenesis is discussed. Mitochondrial DNA mutations can initiate a cascade of events leading to a continuous increase in the production of reactive oxygen species (persistent oxidative stress), a condition that probably favors tumor development. PMID- 12094551 TI - DNA topoisomerase II as a target for cancer chemotherapy. PMID- 12094552 TI - Breast cancer in the setting of HIV infection: cause for concern? PMID- 12094553 TI - Is there life after methotrexate? PMID- 12094554 TI - Is there a role for radionuclide imaging in breast cancer screening? PMID- 12094555 TI - Topoisomerase II inhibitors--teaching old dogs new tricks. PMID- 12094556 TI - The Jewish view of healing. PMID- 12094557 TI - Hypersomnolence and narcolepsy; a pragmatic diagnostic neurophysiological approach. AB - Out of a group of 250 consecutive patients who were examined for various disorders of sleep and waking at Ghent University Hospital within a period of 24 months, 30 patients with hypersomnolence associated with a suspected underlying neurological etiology were selected. The population consisted of 15 males and 15 females with mean age of 36 years (range: 16-60 years). Twenty-one patients had had hypersomnolence for more than 2 years. All patients underwent a single night polysomnography (PSG) and a 4-nap multiple sleep latency test (MSLT). PSG was normal in 23 patients. Sleep onset REM period (SOREMP) was defined as the occurrence of REM sleep within 15 min. after initiation of sleep. PSG demonstrated SOREMP's in only 1 patient and showed evidence of obstructive sleep apnea in 4 patients. Two patients had a low sleep efficiency. MSLT demonstrated hypersomnolence in 17 patients of whom 6 showed SOREMP. Significant hypersomnolence was defined as a mean sleep latency < or = 5 min. 4 patients fulfilled the classical clinical and polygraphic criteria (> or = 2 SOREMP) of narcolepsy. In 8 patients the tentative diagnosis of idiopathic CNS hypersomnolence was made. 13 patients did not sleep during MSLT. These results emphasize the relative importance of MSLT. Our limited 4-nap MSLT protocol proved useful in distinguishing narcolepsy from idiopathic CNS hypersomnolence. PMID- 12094558 TI - Recurrent complex partial status epilepticus associated with tiagabine rechallenge. PMID- 12094559 TI - Neuropathology of human and experimental TSP/HAM: a critical review. AB - Tropical Spastic Paraparesis/HTLV-1 Associated Myelopathy (TSP/HAM) is clinically characterized by chronic insidious spastic paraparesis associated with variable sensory impairment and sphincter symptoms. Neuropathological studies of this condition are based on a few autopsied cases, and on experimental animal models. However, divergent aspects exist between human and experimental animal neuropathology of TSP/HAM, namely, the site of lesions in the spinal cord, the involvement of peripheral nerves and roots, the nature of histological abnormalities, and the cellular reactions. Moreover, unanswered questions as to the preferential site of spinal affection, the temporal inflammatory picture, the selective damage of the corticospinal tract, the sparing of lower motor neurons, the inconsistent affection of sensory tracts, and the involvement of the brain, are outlined. A long-term, chronological, correlated clinical and neuropathological study in HAM experimental animals is suggested. PMID- 12094560 TI - Neuropathology of some hereditary conditions affecting central and peripheral nervous system. AB - Neuropathology plays a crucial role in the phenotypic individualization of hereditary disorders affecting the central and peripheral nervous system even if molecular genetics represents the most essential step in describing the genotypes. The neuropathological description of phenotypes and genotypes can be used for refining clinical skills and understanding many clinical, neurophysiological and neuroradiological features. It contributes to the diagnosis of such disorders. The use of immunohistochemical techniques in combination with molecular genetics improves also our knowledge of their pathogenesis and might participate to the future development of therapeutic strategies. We discuss new features of spino-cerebellar ataxia (SCA) type 7 and of a recently identified SCA17 in order to illustrate the significance of the neuronal intranuclear inclusions (NIIs) described in various CAG/polyglutamine repeat expansion diseases. In the field of the peripheral neuropathies we present data on a newly described autosomal recessive Charcot-Marie-Tooth disease (CMT4F) with mutations in the periaxin gene. We document a dysjunction between myelin loops and axolemma with disappearance of the septate-like junctions or transverse bands. The significance of this dysjunction is not yet elucidated. We hope to show by these examples that the combination of classical and new neuropathological methods is useful in the study of hereditary disorders of the nervous system. PMID- 12094561 TI - Cerebral arteriovenous malformation mimicking acute coronary syndrome. AB - We report a case of cerebral arteriovenous malformation in a 34 year-old male patient presenting with chest pain. Electrocardiographic findings showed ST elevations in the precordial leads. However, a coronary angiogram showed no coronary lesions. Laboratory tests suggested that troponin and cardiac enzymes were within normal limits. Further investigation led to the diagnosis of a cerebral arterio-venous malformation occupying the left temporal and parietal lobes. The chest pain may be explained by the occurrence of a complex partial seizure. PMID- 12094562 TI - Progressive external ophthalmoplegia and multiple mitochondrial DNA deletions. AB - Progressive external ophthalmoplegia (PEO) with secondary accumulation of multiple deletions of mitochondrial DNA (mtDNA) clinically resembles disorders due to primary mutations of mtDNA but follows a Mendelian inheritance pattern. The disorder belongs to an interesting group of diseases in which both the nuclear and the mitochondrial genome are involved in the pathology. Both autosomal dominant (adPEO) and recessive (arPEO) variants of this disorder occur. Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) patients may have multiple mtDNA deletions and/or depletion of mtDNA. Recent reports of mutations in Thymidine Phosphorylase in MNGIE, and of mutations in adenine nucleotide translocator (ANT1), Twinkle and mitochondrial DNA polymerase gamma (POLG) in adPEO, have lead to new insights in the pathogenesis of these disorders of mtDNA maintenance. We also identified POLG mutations in two families with arPEO, which underlines the crucial role of the mtDNA replication machinery for mtDNA maintenance. PMID- 12094563 TI - Therapeutic strategies in the choice of antiepileptic drugs. AB - The choice of treatment of newly diagnosed epilepsy involves many factors such as age, sex, life style, general health and concomitant medication. The seizure type, syndrome, and the pharmacology, efficacy and safety of the antiepileptic drugs (AEDs) should also be considered. Some of the new AEDs appear to provide at least equivalent efficacy with better tolerability. Some of these drugs have the potential to become drugs of first choice in newly diagnosed epilepsy. At the present time, we also must consider the criteria of reimbursement of these drugs. In this paper, we try to describe common and practical strategies to start a treatment of newly diagnosed epilepsy. PMID- 12094564 TI - Micelles and liposomes in metabolic enzyme and glycolipid glycosyltransferase assays. PMID- 12094565 TI - Liposomes and phospholipid binding proteins in glycoprotein biosynthesis. PMID- 12094566 TI - Therapeutic uses of antioxidant liposomes. PMID- 12094567 TI - Targeted gene delivery by virosomes. PMID- 12094568 TI - Preparation and use of liposomes for the study of sphingolipid segregation in membrane model systems. PMID- 12094569 TI - Liposomes containing ligands. Binding specificity to selectins. PMID- 12094570 TI - Preparation and characterization of glycolipid-bearing multilamellar and unilamellar liposomes. PMID- 12094571 TI - Use of liposomes containing carbohydrates for production of monoclonal antibodies. PMID- 12094572 TI - Inhibition of tumor metastasis by liposomes containing glyco-replica peptides. PMID- 12094575 TI - Peptide-induced fusion of liposomes. PMID- 12094574 TI - Preparation, isolation, and characterization of liposomes containing natural and synthetic lipids. PMID- 12094573 TI - Use of phospholipid bilayers and monolayers in binding studies of vitamin K dependent blood coagulation proteins. PMID- 12094576 TI - Liposomes. Applications in protein-lipid interaction studies. PMID- 12094577 TI - Lipids in viral fusion. PMID- 12094578 TI - Liposome-mediated, fluorescence-based studies of sphingolipid metabolism in intact cells. PMID- 12094579 TI - Introduction of dominant inhibitory proteins directed against ROK and MRCK kinases. PMID- 12094580 TI - Effects of Rho family GTPases on cell-cell adhesion. PMID- 12094581 TI - Cell motility and invasion assays. PMID- 12094582 TI - Preparation of GTPases for structural and biophysical analysis. PMID- 12094583 TI - Isolation of regulatory proteins for the Rab3 subfamily GTPases. PMID- 12094584 TI - Analysis and preparation of stable complexes between Rab GTPases, Rab escort protein, and Rab geranylgeranyl transferase. PMID- 12094585 TI - Preparation of myristoylated Arf1 and Arf6 proteins. PMID- 12094587 TI - Arf6 and its role in cytoskeletal modulation. PMID- 12094586 TI - ARF-directed guanine-nucleotide-exchange (GEP) proteins. PMID- 12094588 TI - ARF GTPase-activating protein 1. PMID- 12094589 TI - The use of permeabilized cell systems to study nuclear transport. PMID- 12094590 TI - Analysis of nuclear protein import and export in vitro using fluorescent cargoes. PMID- 12094591 TI - Characterization of the effects of RanGTP on the microtubule cytoskeleton. PMID- 12094592 TI - Using cDNA-representational difference analysis (cDNA-RDA) in combination with microarrays to identify Rac regulated genes. PMID- 12094593 TI - The GTPase cycle. How dominant inhibitory mutants block the biological functions of small GTPases. PMID- 12094595 TI - Ras and Rac as activators of reactive oxygen species (ROS). PMID- 12094594 TI - Fluorescence methods in the study of small GTP-binding proteins. PMID- 12094596 TI - Interfering with Ras signaling using membrane-permeable peptides or drugs. PMID- 12094598 TI - Determination of the activity of Rho-like GTPases in cells. PMID- 12094597 TI - A Ras-based module to generate 32P-labeled fusion proteins for blot overlays. PMID- 12094599 TI - Introduction to thyroid hormone receptors. PMID- 12094600 TI - Gene regulation by thyroid hormone receptors. PMID- 12094601 TI - Molecular analysis of human resistance to thyroid hormone syndrome. PMID- 12094602 TI - Null mutant mice for thyroid hormone receptors. PMID- 12094603 TI - Function of thyroid hormone receptors during amphibian development. PMID- 12094604 TI - Transcriptional regulation by thyroid hormone receptor in chromatin. PMID- 12094605 TI - Gene activation by thyroid hormone receptor in vitro and purification of the TRAP coactivator complex. PMID- 12094606 TI - Transgenic targeting of a dominant negative corepressor to liver and analyses by cDNA microarray. PMID- 12094607 TI - The role of thyroid hormone receptors in the heart. PMID- 12094608 TI - Analysis of thyroid hormone-dependent genes in the brain by in situ hybridization. PMID- 12094609 TI - The v-erbA oncogene. Assessing its differentiation-blocking ability using normal chicken erythrocytic progenitor cells. PMID- 12094610 TI - Molecular targets for nutrients involved with cancer prevention. AB - Dietary nutrients can influence cancer risk by inhibiting or enhancing carcinogenesis through diverse mechanisms of action. The identification and elucidation of their sites of action have been a focus of nutrition and cancer research for more than four decades. Transforming nutrition and cancer research from a predominantly observational to a molecular approach offers exciting opportunities for truly identifying those who will and will not benefit from dietary intervention strategies. The emerging field of nutritional genomics, defined here as the study of any genetic or epigenetic interaction with a nutrient, will be key to this evolution. Unraveling which genetic upregulation or downregulation leads to subsequent phenotype changes will not be easy. There is evidence that genetic polymorphisms can influence the dynamics between nutrients and molecular targets and, thus, contribute to variation in response among individuals. Because many molecular targets will likely be identified, it may be necessary to credential nutrients, that is, to determine which specific nutrient related genetic and epigenetic changes bring about phenotypic changes, to establish which interactions are the most important and under what circumstances. Vitamin D, calcium, folate, selenium, genistein, and resveratrol are highlighted, because they represent specific classes of nutrients and illustrate the need to credential various nutrients to understand their physiological significance in cancer prevention. As the science of nutrition unfolds, a clearer understanding will emerge about how nutrients can modulate cancer risk through molecular interactions and how foods might be changed by agronomic approaches and/or biotechnology. Undeniably, embracing new genomic technologies offers exciting opportunities for advances in the broad area of nutrition, especially those related to cancer prevention. PMID- 12094611 TI - Conjugated linoleic acid does not inhibit development of aberrant crypt foci in colons of male Sprague-Dawley rats. AB - Conjugated linoleic acid (CLA) is a potent inhibitor of the initiation and promotion of mammary carcinogenesis in animal models, but its role in colon carcinogenesis remains unclear. The objective of this study was to determine whether CLA inhibits the promotion of colon carcinogenesis. Forty male Sprague Dawley rats were given a single dose of azoxymethane (20 mg/kg body wt i.p.). After 1 wk, the animals were randomized into two groups (n = 20) and fed a control AIN-93G diet or the control diet supplemented with 1% CLA at the expense of the soybean oil. After 12 wk, the animals were killed, and their colons were stained with methylene blue for aberrant crypt foci (ACF) analysis by light microscopy. The total number of ACF per animal did not differ between the control (174 +/- 11) and CLA (170 +/- 10) groups. Furthermore, CLA did not affect the average crypt multiplicity (crypts/ACF) or the average number of ACF in any size category. However, rats fed the 1% CLA diet had significantly higher serum insulin levels at the time of sacrifice than those fed the control diet. Thus it is possible that the promoting effects of elevated serum insulin on colon carcinogenesis may have counteracted an inhibitory effect of CLA. PMID- 12094612 TI - Effect of simple phenolic compounds on azoxymethane-induced aberrant crypt foci in rat colon. AB - Because complex mixtures of plant polyphenols exert anticancer activity in animal models, we investigated whether low-molecular-weight natural phenolic compounds (2-OH-coumaric acid, 3-OH-coumaric acid, 4-OH-coumaric acid, 3-OH-flavone, 7-OH flavone, 4-OH-benzoic acid, 3-OH-benzoic acid, and 2,3-OH-benzoic acid) affect azoxymethane (AOM)-induced aberrant crypt foci (ACF), which have been suggested to represent preneoplastic lesions, in the colon of rats. Male Fischer 344 rats were fed diets supplemented with 0.1% (wt/wt) of the different phenolic compounds, and after 2 wk they were treated twice (1 wk apart) with AOM (15 mg/kg s.c.); the dietary treatment continued until sacrifice, 7 wk after the first injection with AOM. The results showed that none of these phenolic compounds exerted chemopreventive activity on the ACF assay. On the contrary, 3-OH-flavone slightly, although significantly, increased (P < 0.05), the number of ACF per colon [157 +/- 7 and 198 +/- 14 (SE) in control and 3-OH-flavone groups, respectively, n = 10]. We also found that the number of "large" ACF was significantly increased in the group treated with 4-OH-benzoic acid. In conclusion, none of the phenolic compounds tested demonstrated a suppressive action on ACF induction by AOM. PMID- 12094613 TI - Response of hypothalamic NPY mRNAs to a negative energy balance is less sensitive in cachectic mice bearing human tumor cells. AB - We selected three human cancer cell lines [human melanoma (SEKI), human melanoma (G361), and human neuroepithelioma (NAGAI)] that have an ability to develop cancer cachexia syndrome with and without accompanying anorexia and examined the hypothalamic levels of mRNAs for neuropeptide Y (NPY), melanin-concentrating hormone, and orexin. The body weight of sham-operated mice continued to increase, while mice of all tumor-bearing groups lost weight. Competitive reverse transcription-polymerase chain reaction analysis showed that, regardless of feeding status, NPY mRNA levels were elevated in all tumor-bearing mice compared with sham-operated mice, although to a lesser degree than weight-matched pair weight mice. Melanin-concentrating hormone and orexin mRNA in the hypothalamus followed the same pattern as NPY, although most of the differences did not reach statistical significance. These results support the notion that the response of NPY mRNA to a negative energy balance is less sensitive in these rodent models of cancer cachexia. PMID- 12094614 TI - Green tea catechins and vitamin E inhibit angiogenesis of human microvascular endothelial cells through suppression of IL-8 production. AB - Epidemiological and animal studies have indicated that consumption of green tea and high vitamin E intake are associated with a reduced risk of developing certain forms of cancer. However, the inhibitory mechanism of green tea catechins and vitamin E in angiogenesis, an important process in tumor growth, has not been well established. In the present study, alpha-tocopherol and several major catechins of green tea (catechin, epicatechin, epicatechin gallate, epigallocatechin, and epigallocatechin gallate) were tested for their ability to inhibit tube formation in vitro using a model in which human microvascular endothelial cells were exposed to a constant rate of a physiologically low level of H2O2. In this model, the production of interleukin (IL)-8 by human microvascular endothelial cells at a low level of H2O2 was required for angiogenesis, as assessed by tube formation in three-dimensional gel in culture. Vitamin E (d-alpha-tocopherol, 40 microM) in the culture media significantly reduced IL-8 production and angiogenesis. Among the green tea catechins, epigallocatechin (0.5-1 microM) was the most effective in reducing IL-8 production and inhibiting angiogenesis. These results suggest that consumption of green tea catechins or supplemental intake of vitamin E may have preventive effects on tumor development, mediated, at least in part, through inhibition of angiogenesis via suppression of IL-8 production. PMID- 12094615 TI - Linoleic acid-induced VCAM-1 expression in human microvascular endothelial cells is mediated by the NF-kappa B-dependent pathway. AB - Vascular cell adhesion molecule-1 (VCAM-1) has been reported to play an important role in cancer metastasis via the adhesive interaction between tumor cells and endothelial cells. In this study, we examined the effects of linoleic acid on VCAM-1 expression and its transcriptional regulatory mechanism in human microvascular endothelial cells (HMEC-1). Time- and dose-dependent increases of VCAM-1 mRNA levels were observed in linoleic acid-treated HMEC-1 as detected by reverse transcriptase-polymerase chain reaction. Flow cytometry analysis showed a significant and dose-dependent upregulation of VCAM-1 expression in HMEC-1 stimulated with linoleic acid compared with controls. To clarify the transcriptional regulatory pathway, we investigated the role of nuclear factor kappa B (NF-kappa B) in the expression of VCAM-1 by linoleic acid in HMEC-1. Nuclear extracts from HMEC-1 stimulated with linoleic acid showed a dose dependent increase in binding activity to the NF-kappa B consensus sequences. These effects were preventable by cotreatment with inhibitors of NF-kappa B activity, such as sodium salicylate, aspirin, or pyrrolidine dithiocarbamate. In addition, pretreatment with NF-kappa B inhibitors markedly suppressed the ability of linoleic acid to induce VCAM-1 gene expression. The role of NF-kappa B in linoleic acid-induced VCAM-1 expression was confirmed by functional promoter studies in HMEC-1 transfected with reporter constructs of the VCAM-1 promoter with or without mutated NF-kappa B binding site. These results indicate that linoleic acid upregulates VCAM-1 expression in HMEC-1 through the NF-kappa B dependent pathway. PMID- 12094616 TI - Carnosic acid inhibits proliferation and augments differentiation of human leukemic cells induced by 1,25-dihydroxyvitamin D3 and retinoic acid. AB - Carnosic acid, the polyphenolic diterpene derived from rosemary, is a strong dietary antioxidant that exhibits antimutagenic properties in bacteria and anticarcinogenic activity in various cell and animal models. In the present study, we show that carnosic acid (2.5-10 microM) inhibits proliferation of HL-60 and U937 human myeloid leukemia cells (half-maximal inhibitory concentration = 6 7 microM) without induction of apoptotic or necrotic cell death. Growth arrest occurred concomitantly with a transient cell cycle block in the G1 phase, which was accompanied by an increase in the immunodetectable levels of the universal cyclin-dependent kinase inhibitors p21WAFI and p27Kipl. Carnosic acid caused only a marginal induction of differentiation, as monitored by the capacity to generate superoxide radicals and the expression of cell surface antigens (CD11b and CD14) and receptors for the chemotactic peptide N-formyl-L-methionyl-L-leucyl-L phenylalanine. However, at low concentrations, this polyphenol substantially augmented (100- to 1,000-fold) the differentiating effects of 1,25 dihydroxyvitamin D3 and all-trans retinoic acid. Furthermore, such combinations of carnosic acid and any of these differentiation inducers synergistically inhibited proliferation and cell cycle progression. These results indicate that carnosic acid is capable of antiproliferative action in leukemic cells and can cooperate with other natural anticancer compounds in growth-inhibitory and differentiating effects. PMID- 12094617 TI - Estrogenic activity of naturally occurring anthocyanidins. AB - Anthocyanins, which are natural plant pigments from the flavonoid family, represent substantial constituents of the human diet. Because some other bioflavonoids are known to have estrogenic activity, the aim of this study was to determine the estrogenic activity of the anthocyanine aglycones. Binding affinity to the estrogen receptor-alpha was 10,000- to 20,000-fold lower than that of the endogenous estrogen estradiol. In the estrogen receptor-positive cell line MCF-7, the anthocyanidins induced expression of a reporter gene. The tested anthocyanidins showed estrogen-inducible cell proliferation in two cell lines (MCF-7 and BG-1), but not in the receptor-negative human breast cancer cell line MDA-MB-231. The phytoestrogen-induced cell proliferation could be blocked by addition of the receptor antagonist 4-hydroxytamoxifen. Combination treatments with the endogenous estrogen estradiol resulted in a reduction of estradiol induced cell proliferation. Overall, the tested anthocyanidins exert estrogenic activity, which might play a role in altering the development of hormone dependent adverse effects. PMID- 12094618 TI - Comparison of calcium supplementation or low-fat dairy foods on epithelial cell proliferation and differentiation. AB - Epidemiological evidence suggests that dietary calcium and vitamin D intake are inversely related to incidence of colon cancer. Previous studies have demonstrated that supplementation of the diet with calcium in the form of calcium tablets or low-fat dairy foods alters colonic epithelial cell proliferation from a higher- to a lower-risk pattern. The present study compared relative effects of administration of calcium carbonate at approximately 900 mg/day (calcium) with those of a low-fat dairy food diet providing about the same amount of calcium (dairy) in a cross-over "head-to-head" study of 40 subjects at risk for colonic neoplasia. Dietary intake of macronutrients was similar in the two study periods, except for a slight increase in protein intake during dairy calcium supplementation. Rectal epithelial cell proliferation was studied in flat endoscopically normal-appearing mucosa at baseline and at the end of each of the two study periods and showed a significant reduction in epithelial crypt cell labeling index from 12.5% to 9.1% (calcium) or 9.3% (dairy) as well as in proliferating cells in the upper 40% of the crypt from 0.09 to 0.03 in the calcium- and low-fat dairy-supplemented intervention groups. No significant changes in two epithelial cell differentiation markers, cytokeratin AE1 and acidic mucins, were found. Furthermore, there were no differences in epithelial cell apoptosis or expression of the proapoptotic gene product BAK. These data indicate that increased dietary calcium given as supplements or in the diet in low-fat dairy foods lowers epithelial cell proliferation indexes from a higher- to a lower-risk pattern. Because supplemental calcium has been shown to reduce the recurrence of colonic adenomatous polyps in patients at increased risk for colonic neoplasia, our data suggest that supplemental low-fat dairy foods may also be effective. PMID- 12094619 TI - Butyrate induces glutathione S-transferase in human colon cells and protects from genetic damage by 4-hydroxy-2-nonenal. AB - Butyrate, one of the major products of gut fermentation, is known to inhibit proliferation, induce apoptosis and differentiation, and increase phase II enzyme activities in tumor cells, whereas little information is available on protective effects in less-transformed colon cells. The aim of this study was to investigate whether the chemoprotective mechanism of glutathione S-transferase (GST) induction by butyrate could also play a role in earlier stages of colon carcinogenesis and whether chemoresistance of cells toward the endogenous genotoxic risk factor 4-hydroxy-2-nonenal (HNE) could be a consequence of butyrate treatment. As cell models, we used the human tumor cell lines HT29 and HT29 clone 19A, a differentiated subclone with properties resembling primary colon cells. We determined the expression of GSTP1 protein (enzyme-linked immunosorbent assay), the major GST in HT29, GSTP1 mRNA (Northern blotting), GST activity, intracellular glutathione, and total protein. The genotoxic impact of HNE (100-200 microM) was compared in butyrate-treated and nontreated cells using single-cell microgel electrophoresis. Our results show that GSTP1 mRNA, GSTP1 protein, GST activity, and total protein were increased (1.2- to 2.5-fold) and glutathione levels were maintained after 24-72 h of incubation with 4 mM butyrate. Moreover, a marked reduction of HNE-induced genotoxicity was caused by preincubation with butyrate. Butyrate also induced the phosphorylation of extracellular signal-regulated kinases (ERK1/2, Western blotting) after 5-30 min, which indicates a regulation of GST expression by this signal pathway. Most effects were greater in HT29 parent cells than in clone cells. In conclusion, butyrate enhances expression of GST and other proteins in both cell lines, which leads to an enhanced chemoprotection, reducing the impact of HNE genotoxicity. Thus butyrate could play a role in early and later stages of cancer prevention by reducing exposure to relevant risk factors. PMID- 12094620 TI - Isoflavones inhibit proliferation of ovarian cancer cells in vitro via an estrogen receptor-dependent pathway. AB - Incidence rates of ovarian cancer remain lowest in Asian nations, which consume diets rich in soy products, whereas they remain among the highest in the United States and other Western nations, which consume low amounts of soy foods. The hypothesis of this study is that soy-derived isoflavones inhibit the proliferation of ovarian cancer cells in vitro by regulating cytokine synthesis. Cell proliferation was evaluated by bromodeoxyuridine and 3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium bromide assay. DNA synthesis of Caov-3 and NIH:OVCAR-3, two ovarian cancer cell lines, was significantly inhibited by genistein or daidzein at dietarily relevant concentrations (10(-8)-10(-10) M). Also, the number of viable cells was significantly lower (45-75%) in all isoflavone-treated groups than in the control group (P < 0.01). The addition of ICI-182780, an estrogen antagonist, blocked these inhibitory effects. In addition, interleukin-6 synthesis by these two cell lines was inhibited by genistein or daidzein; production was decreased by approximately 20% compared with the control group (P < 0.05). In contrast, transforming growth factor-beta 1 production in ovarian cancer cells incubated with genistein or daidzein was significantly greater, i.e., by approximately 30%, than in the control group (P < 0.05). Addition of ICI-182780 also neutralized the effects of isoflavones on the production of these two cytokines by ovarian cancer cells. In summary, genistein and daidzein independently modify cytokine production and reduce ovarian cancer cell proliferation via, at least in part, an estrogen receptor-dependent pathway. PMID- 12094621 TI - Cruciferous vegetables and cancer prevention. AB - In recent years, cancer prevention by natural products has received considerable attention. The potential protective role of cruciferous vegetables and active components present in these vegetables, such as isothiocyanates and indole-3 carbinol, has been extensively studied in experimental in vitro and in vivo carcinogenesis models. Results have consistently shown that the chemopreventive agents derived from this class of vegetables of the Cruciferae family influence carcinogenesis during initiation and promotion phases of cancer development. Similarly, reports from epidemiological studies and clinical trials support this notion. However, there is no comprehensive summary of all these aspects of the association between cruciferous vegetables and cancer prevention. We have attempted to summarize experimental carcinogenesis studies as well as clinical trials and studies on the mechanism of action of selective chemopreventive agents isolated and identified within these natural products. Results clearly point toward a positive correlation between cancer prevention of many target organs and consumption of cruciferous vegetable or their active constituents. Yet we are still far from complete understanding of the effects of combinations of chemopreventive phytochemicals present in these cruciferous vegetables and their overall mechanism(s) of action in providing protective effects. PMID- 12094622 TI - Neurotensin-and EGF-induced metabolic activation of colon carcinoma cells is diminished by dietary flavonoid cyanidin but not by its glycosides. AB - Dietary polyphenols, including anthocyanidins and their glycosides anthocyanins, are suggested to be involved in the protective effects of fruits and vegetables against cancer. Very few data are available concerning the effects of anthocyanidins/anthocyanins on cellular processes induced by growth factors such as neurotensin and epidermal growth factor (EGF), which are implicated in the pathophysiology of colon cancer. Here, we show that neurotensin and EGF caused an increase in the extracellular acidification rate, which could reflect the activity of cellular metabolism, in the human carcinoma cell line HT29 clone 19A. Neurotensin and EGF also caused a strong rise in the intracellular Ca2+ concentration, induced phosphorylation of extracellular signal-regulated kinases (ERK1 and ERK2), and stimulated growth of human carcinoma cells. Cyanidin (10 microM), but not its glycosides cyanin and idaein, was able to inhibit the neurotensin- and EGF-induced increased rate of extracellular acidification. In contrast to N-ethyl-N-isopropyl amiloride, an inhibitor of Na+/H+ exchange, cyanidin did not alter the rate of intracellular pH recovery of cells loaded by NH3/NH4+, indicating that cyanidin inhibits cellular metabolism, rather than directly altering Na+/H+ exchange. Cyanidin, but not cyanin and idaein, was able to inhibit an increase in intracellular Ca2+ concentration induced by neurotensin. Neurotensin- and EGF-induced phosphorylation of ERKs was not affected by cyanidin, cyanin, and idaein at < or = 100 microM. Only cyanidin (100 microM), but not cyanin and idaein, was able to inhibit cellular growth induced by EGF. Thus these findings suggest that a dietary polyphenol cyanidin, but not its glycosides, is a potent inhibitor of mitogen-induced metabolic activity, increase in free intracellular Ca2+, and cellular growth of cultured colon carcinoma cells. PMID- 12094623 TI - Abrogation of estrogen-mediated cellular and biochemical effects by indole-3 carbinol. AB - The use of naturally occurring phytoantiestrogens for prevention and therapy of breast cancer is an alternative to synthetic antiestrogens. We have been examining the mechanism of action of the antiestrogen indole-3-carbinol (I3C), a constituent of compounds present in cruciferous vegetables. I3C abrogates the cell-proliferative effect of 17 beta-estradiol (E2), as observed in several different estradiol-responsive breast cancer cell lines and isolated cell clones. Modulation of E2 activity by I3C, in part, was by the induction of the 2 hydroxylation pathway, one of the two competing hydroxylation pathways of estrone conversion that resulted in the formation of metabolites with antiestrogenic properties. I3C-mediated induction of the 2-hydroxylation pathway correlated with a selective induction of cytochrome P-450 1A1 by I3C in E2-responsive human breast cancer cells. Induction of neither the 2-hydroxylation pathway nor cytochrome P-450 1A1 was observed in estrogen-nonresponsive human breast cancer cells. This selective effect warranted a further search for biochemical targets of I3C related to E2 function. To this end, we observed that E2-mediated phosphorylation of the estrogen receptor is inhibited by I3C. Our results are consistent with the hypothesis that I3C exerts its antiestrogenic effect by intervention in the E2-estrogen receptor signal transduction pathways and by alterations in E2 metabolism that resulted in the formation of metabolites with antiestrogenic activity. PMID- 12094625 TI - Vitamin/mineral supplementation among cancer survivors: 1987 and 1992 National Health Interview Surveys. AB - The number of cancer survivors in the United States is increasing, but little is known about this population, including its use of vitamin/mineral supplements. We combined data on vitamin/mineral use from the 1987 and 1992 National Health Interview Survey Cancer Epidemiology Supplement (CES) for cancer survivors: persons reporting a diagnosis of cancer other than nonmelanoma skin cancer > 5 yr before their interviews [n = 461 (1987) and 228 (1992)] and persons reporting no history of cancer [n = 20,851 (1987) and 11,186 (1992)]. For both groups, we calculated gender-specific proportions (adjusted for age, race/ethnicity, education, smoking status, and poverty index) for use of multivitamins, vitamins A, C, and E, and calcium during the year before each survey. Supplement use was similar in survivors and persons reporting no history of cancer. Among survivors, calcium use was significantly higher among women (34.9%) than men (13.8%), and vitamin A use was higher among men than women (P < 0.05). Over three-fourths of both groups used multivitamins, and about one-half used vitamin C. No differences were found in vitamin/mineral use between male survivors and men with no cancer history or between female survivors and women with no cancer history. These first nationally representative estimates suggest that persons who have survived cancer and those who report that they never had the disease do not differ appreciably in their use of vitamin/mineral supplements. Results were based on small numbers of survivors, however, and require replication. PMID- 12094624 TI - Prostaglandin E2 is involved in the increase of cytochrome P-450 2B1 expression by alpha-tocopheryl succinate in primary rat hepatocytes in the presence of phenobarbital. AB - The modulation of cytochrome P-450 2B1 expression by alpha-tocopheryl succinate and whether prostaglandin E2 is involved in this modulation in primary rat hepatocytes in the presence of phenobarbital were investigated. A primary rat hepatocyte culture model that faithfully reproduces the phenobarbital response observed in vivo was used. Intracellular alpha-tocopherol content was dose dependently increased by alpha-tocopheryl succinate incubation. Hepatocytes were demonstrated to have prostaglandin E2-synthesizing capability. alpha-Tocopheryl succinate inhibited prostaglandin E2 synthesis by hepatocytes and increased cytochrome P-450 2B1 expression in the presence of phenobarbital; however, it had little effect on intracellular cAMP level. To mimic the exogenous source of prostaglandin E2 from nonparenchymal cells, various concentrations of prostaglandin E2 were added to the cell culture. High doses of exogenous prostaglandin E2 (100 and 1,000 nM) inhibited the cytochrome P-450 2B1 expression in the presence of phenobarbital compared with low doses (1 and 10 nM); however, the presence of high doses of prostaglandin E2 had no effect on intracellular cAMP level. Forskolin significantly increased intracellular cAMP level and inhibited cytochrome P-450 2B1 expression in the presence of phenobarbital. The results of this study indicate that alpha-tocopheryl succinate increases cytochrome P-450 2B1 expression via its inhibition of prostaglandin E2 synthesis in the presence of phenobarbital; however, changes in intracellular cAMP level are not related to cytochrome P-450 2B1 expression. PMID- 12094626 TI - Tea consumption and risk of cancer of the colon and rectum. AB - The association between tea consumption and risk of colon and rectal cancers was investigated in a population-based case-control study conducted in Iowa (United States). Colon (n = 685) and rectal (n = 655) cancer cases age 40-85 yr were identified through the Iowa Surveillance, Epidemiology, and End Results (SEER) Cancer Registry (86% response rate); controls (n = 2,434) were frequency matched by sex and 5-yr age group (80% response rate). The usual adult consumption of tea (hot and iced), along with other information including dietary data, was self reported using a mailed questionnaire. Total tea consumption (cups/day) was categorized as none (reference category), low (< 3.1), medium (3.1-5.0), and high (> 5.0), with cut points for tea consumers based on the 75th and 90th percentiles of use among controls. Unconditional logistic regression was used to estimate the odds ratios (ORs) and 95% confidence intervals. There was no association between total tea consumption and colon cancer (ORs = 1.0, 1.1, 1.3, and 0.7) or rectal cancer (ORs = 1.0, 0.9, 1.4, and 1.0) after adjustment for age, sex, education, physical activity, smoking history, and intake of coffee, fiber, and fruits and vegetables. Results were similar when hot tea and iced tea were evaluated individually. Further adjustment for other colorectal cancer risk factors did not alter these results. There was no association with proximal or distal colon cancer. There was also no interaction between tea consumption and any of the dietary variables or total fluid on risk of colon or rectal cancer, with the exception of a suggestive positive association between an increasing frequency of tea consumption and colon cancer risk among current smokers (multivariate ORs = 1.0, 1.4, 2.0, and 1.8; P for trend = 0.1), but not among never smokers (multivariate ORs = 1.0, 1.0, 1.1, and 0.4; P for trend = 0.3). These data do not support an overall association, either positive or negative, between tea consumption and risk of colon or rectal cancer in this Mid-western US population. PMID- 12094627 TI - Soy milk intake in relation to serum sex hormone levels in British men. AB - Soy beans contain high levels of the isoflavones genistein and daidzein and their glycosides and have been implicated in the prevention of prostate cancer, possibly via their effects on sex hormone metabolism. The aim of this study was to assess the relation between dietary soy intake and sex hormone levels in a cross-sectional analysis of 696 men with a wide range of soy intakes. Soy milk intake was measured using a validated semiquantitative food frequency questionnaire, and serum hormone concentrations were measured by immunoassay. Multiple regression was used to investigate the association between soy milk intake, an index of isoflavone intake, and hormone levels after adjustment for pertinent confounders. Soy milk intake was not associated with serum concentrations of testosterone, free testosterone, androstanediol glucuronide, sex hormone-binding globulin, or luteinizing hormone. These results suggest that soy milk intake, as a marker of isoflavone intake, is not associated with serum sex hormone concentrations among free-living Western men. PMID- 12094628 TI - Sex hormone levels in premenopausal African-American women with upper and lower body fat phenotypes. AB - Body fat distribution may be a better marker of a hormonal pattern associated with increased breast cancer risk than obesity. This cross-sectional study of 106 healthy premenopausal African-American (AA) women compared the midfollicular phase sex hormone and sex hormone-binding globulin levels in upper body fat (UBF) and lower body fat (LBF) phenotype and obese and nonobese women. Multivariate regression analyses were used to control for various confounders, including dietary factors. UBF phenotype women had 37% (P = 0.02), 50% (P = 0.01), 52% (P = 0.007), and 50% (P = 0.009) higher levels of estradiol (E2), free E2, testosterone (T), and free T, respectively, than LBF phenotype women. Only %free T was higher in obese than in nonobese women (P = 0.02). The levels of E2, free E2, %free E2, T, and free T were higher [by 42% (P = 0.01), 68% (P = 0.001), 18% (P = 0.04), 36% (P = 0.04), and 61% (P = 0.01), respectively] and the level of sex hormone-binding globulin was lower [by 28% (P = 0.04)] in obese UBF than in nonobese LBF phenotype women. These findings support the hypothesis that body fat distribution may be a better marker of a hormonal pattern associated with increased breast cancer risk than obesity. Obese UBF phenotype AA women, in particular, have a high-risk hormonal profile. Future breast cancer studies might consider controlling for measures of obesity and body fat distribution to minimize confounding. PMID- 12094629 TI - Quantitative determination of 3,3'-diindolylmethane in urine of individuals receiving indole-3-carbinol. AB - Indole-3-carbinol (I3C) or, more correctly, its acid condensation products act as chemoprotective agents via several mechanisms. It induces the expression of cytochrome P-450 1A1, which shifts the estrogen metabolic pathway in favor of C-2 hydroxylation and away from the formation of 16 alpha-hydroxyestrone, a suspected endogenous carcinogen. Increased 16 alpha-hydroxylation of estrogen is associated with greater risk of cancer of the cervix, breast, endometrium, and larynx. The production of 4-hydroxyestrone is also inhibited by I3C. I3C can induce a G1 cell cycle arrest in human MCF-7 breast cancer cells. It can suppress aberrant crypt foci. I3C significantly inhibits the cell adhesion, spreading, and invasion associated with an upregulation of PTEN (a tumor suppressor gene) and E-cadherin (a regulator of cell-cell adhesion) expression in T47-D human breast cancer cells. Thus I3C exhibits anticancer activities by suppressing breast tumor cell growth and metastatic spread. A gas chromatography-mass spectrometry method for the quantitation of diindolylmethane, the principal acid condensation product of I3C, has been developed for use in determining compliance in subjects who have been treated with I3C. The method utilizes a 1-ml urine sample. We have used this method to correlate I3C ingestion with regression of cervical intraepithelial neoplasia in a population of women at risk for cervical cancer. The assay provides an objective marker of consumption using a noninvasive biological fluid and illustrates that diindolylmethane may be used as a marker of compliance in I3C dietary intervention studies. PMID- 12094630 TI - Measurement of the systemic inflammatory response predicts cancer-specific and non-cancer survival in patients with cancer. AB - The assessment of prognosis in patients with advanced cancer remains problematical. The value of C-reactive protein concentration in this context has not been clearly defined. Patients with a diagnosis of colorectal (n = 182), gastric (n = 87), breast (n = 99), or bronchogenic (n = 404) cancer and who had measurements of C-reactive protein and albumin were identified. Median survival, from the time of sampling, ranged from 478 days in the colorectal cancer patients to 60 days in patients with bronchogenic cancer. On univariate analysis, there was, in each tumor type, a significant relationship between the duration of survival and both log10 C-reactive protein and albumin concentrations (P < or = 0.0002). On multivariate analysis, in each tumor type, log10 C-reactive protein remained a significant independent predictor of survival (P < or = 0.0002). When all four groups of cancer patients were analyzed (n = 772), the hazard ratio for a 10-fold increase in C-reactive protein concentration in cancer-specific survival was 2.21 (95% confidence interval = 1.92-2.56, P < 0.0001) and the corresponding hazard ratio for non-cancer survival was 5.48 (95% confidence interval = 3.55-8.46, P < 0.0001). The results of the present study indicate that in advanced cancer patients the presence of a systemic inflammatory response and the magnitude of that response predict the duration of cancer-specific and non cancer survival. PMID- 12094631 TI - Increasing dietary supervision can reduce weight loss in oral cancer patients. AB - Weight loss after treatment for intraoral malignancy is common and can impair patient recovery and wound healing. We report a reduction in weight loss in patients undergoing combined-modality treatment (surgery + radiotherapy) after an increase in dietary supervision and changes to the patient dietary protocol. Two groups of patients were compared: Group I received treatment before the protocol changes, and Group II received treatment after the changes took effect. After surgery the average weight loss for Group I was 3.67% compared with 2.42% for Group II (P < 0.05), after radiotherapy the average weight loss for Group I was 6.56% compared with 4.83% for Group II (P < 0.05), and after combined-modality treatment the average weight loss for Group I was 9.83% compared with 6.6% for Group II (P < 0.05). The successful protocol changes included increased supervision of patients by the dedicated head-and-neck team dietitian when patients were undergoing radiotherapy, including the period between completion of surgery and commencement of radiotherapy. The period of time spent without specialist dietary supervision was therefore reduced. Specific guidelines were devised to allow for increased dietary input when required, specifically, if patients lost weight for > or = 2 wk or if they were being fed enterally. These changes were made after an audit that demonstrated that severe weight loss in this group of patients was common. We have demonstrated that regular appraisal of weight loss in this group of patients is valuable and that this weight loss can be reduced if appropriate action is taken. PMID- 12094632 TI - Chemopreventive effects of soy protein and purified soy isoflavones on DMBA induced mammary tumors in female Sprague-Dawley rats. AB - There are conflicting reports on the effect of soy and its components on mammary carcinogenesis in adult female rats, mainly because of different rodent models that are used in chemoprevention studies. The present study was undertaken to compare the tumor-preventative effects of soy protein isolate (SPI) and two of its isoflavones in a "standard" model that had been used for the identification of many chemopreventive agents. Six groups of female Sprague-Dawley rats were provided with modified cornstarch AIN-76A diets supplemented as follows: no additional agents (control), purified genistein (200 mg/kg diet), purified daidzein (200 mg/kg diet), genistein + daidzein (100 mg/kg diet each), SPI containing normal levels of isoflavones (SPI-n), or SPI depleted of isoflavones (SPI-d). Mammary carcinomas were induced by 7,12-dimethylbenz[a]anthracene (DMBA) introduced 1 wk after the animals began consuming the experimental diets. At the end of the study (120 days after DMBA treatment), no significant differences were found among the six groups with respect to tumor incidence or survival, nor was there a significant reduction in tumor multiplicity in the genistein or genistein + daidzein group. However, there was a 32% reduction in tumor multiplicity in the daidzein and SPI-n groups relative to the control group (P < 0.05). The most effective diet was SPI-d, which produced a 50% reduction in tumor multiplicity relative to the control (P < 0.01). The difference between the SPI-d group and the daidzein or SPI-n group was not significant. Median tumor latency was increased from 53 days in the control group to 68 days in the daidzein group and to 72 days in the SPI-d group, but these differences were not statistically significant. These results show that daidzein and SPI (with normal or low levels of isoflavones) are effective inhibitors of DMBA-induced mammary tumors in adult rats. PMID- 12094633 TI - Uptake and metabolism of hydroxymatairesinol in relation to its anticarcinogenicity in DMBA-induced rat mammary carcinoma model. AB - The chemopreventive effects of hydroxymatairesinol (HMR), a lignan extracted from Norway spruce (Picea abies), on the development of mammary carcinoma induced by 7,12-dimethylbenz[a]anthracene (DMBA) was studied in rats. HMR administered via diet in an average daily dose of 4.7 mg/kg body wt starting before DMBA induction reduced tumor volume and tumor growth, but no significant reduction in tumor multiplicity (number of tumors/rat) was observed. The predominant histological type in the control group was type B (well-differentiated adenocarcinoma, 78%). The proportion of type B tumors decreased to 35% in the HMR group, while the type A (poorly differentiated) and type C (atrophic) tumor proportions increased. Anticarcinogenic effects of dietary HMR (4.7 mg/kg) were also evident when the administration started after DMBA induction and was seen as growth inhibition of established tumors. Dietary HMR supplementation significantly increased serum and urinary enterolactone and HMR concentrations but had no significant effect on the uterine weight, suggesting that HMR or its major metabolite enterolactone did not have an antiestrogenic effect. Further studies are warranted to further clarify and verify HMR action and the associated mechanisms in mammary tumorigenesis. PMID- 12094634 TI - Vaccenic acid feeding increases tissue levels of conjugated linoleic acid and suppresses development of premalignant lesions in rat mammary gland. AB - The objective of this report was to determine whether vaccenic acid (t11-18:1) is converted efficiently to conjugated linoleic acid (c9,t11-18:2, CLA) in rats via the delta 9-desaturase reaction and, if so, whether vaccenic acid could substitute for CLA as an anticancer agent. In Study 1, rats were fed 1%, 2%, or 3% vaccenic acid in their diet, and tissue levels of CLA and CLA metabolites were determined in liver and mammary gland. In general, concentrations of CLA and CLA metabolites increased proportionately with an increase in vaccenic acid intake, at least up to the 2% dose level. Beyond this dose, there was clearly a plateauing effect. Thus vaccenic acid concentration increased from an undetectable level in the control to 78.5 nmol/mg lipid in the liver of rats fed a 2% vaccenic acid diet. This was accompanied by an increase in CLA from 2.3 to 33.6 nmol/mg lipid. These changes were also mirrored in the mammary gland, where increases in vaccenic acid (from 27.5 to 163.2 nmol/mg lipid) and CLA (from 17.8 to 108.9 nmol/mg lipid) were similarly observed. Vaccenic acid at 2% produced a CLA concentration in the mammary gland that was historically associated with a positive response in tumor inhibition based on our past experience. This provided the basis for selecting 2% vaccenic acid in Study 2, which was designed to evaluate its efficacy in blocking the development of premalignant lesions in the rat mammary gland. In this experiment, formation of histologically identifiable pathology due to intraductal proliferation of terminal end bud cells of mammary epithelium was used as the end point of analysis at 6 wk after carcinogen administration. Treatment with vaccenic acid reduced the total number of these premalignant lesions by approximately 50%. We hypothesize that the anticancer response to vaccenic acid is likely to be mediated by its endogenous conversion to CLA via delta 9-desaturase. PMID- 12094636 TI - [Remarks on the olfactory perception mechanism]. AB - After presenting the morphology of the olfactory epithelium, the author describe the smell stimulation mechanism of the olfactory receptor. The importance of the connection between the receptor and protein G is emphasised. The author also presents the theories on the possibility of smell identification in the peripheral olfactory system. PMID- 12094635 TI - Inhibition by white tea of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine induced colonic aberrant crypts in the F344 rat. AB - There is growing interest in the potential health benefits of tea, including the anticarcinogenic properties. We report here that white tea, the least processed form of tea, is a potent inhibitor of 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP)-induced colonic aberrant crypts in the rat. Male Fischer 344 rats were treated for 8 wk with white tea (2% wt/vol) or drinking water alone, and on alternating days in experimental Weeks 3 and 4 the animals were given PhIP (150 mg/kg body wt p.o.) or vehicle alone. At the end of the study there were 5.65 +/- 0.81 and 1.31 +/- 0.27 (SD) aberrant crypt foci per colon in groups given PhIP and PhIP + white tea, respectively (n = 12, P < 0.05). No changes were detected in N-acetyltransferase or arylsulfotransferase activities compared with controls, but there was marked induction of ethoxyresorufin O-deethylase, methoxyresorufin O-demethylase, and UDP-glucuronosyltransferase after treatment with white tea. Western blot revealed corresponding increases in cytochrome P-450 1A1 and 1A2 proteins. Enzyme assays and Western blot also revealed induction of glutathione S-transferase by white tea. There was less parent compound and 4' hydroxy-PhIP but more PhIP-4'-O-glucuronide and PhIP-4'-O-sulfate in the urine from rats given PhIP + white tea than in urine from animals given carcinogen + drinking water. The results indicate that white tea inhibits PhIP-induced aberrant crypt foci by altering the expression of carcinogen-metabolizing enzymes, such that there is increased ring hydroxylation at the 4' position coupled with enhanced phase 2 conjugation. PMID- 12094637 TI - [Gene modified tumor vaccines in therapy of malignant melanoma]. AB - Over the years the incidence of malignant melanoma in Poland as well as in other countries has been continuously increasing. Surgery is a treatment of choice in the early stages of primary lesions. Advanced malignant melanoma however is resistant to chemotherapy or radiotherapy. Therefore there is a need for new, more effective treatments. In the last years biotherapy such as immunotherapy is focusing a lot of attention. Unfortunately, systemic administration of immunostimulatory factors is very often associated with severe side effects. Thus, concepts of specific immunotherapies such as immunogene therapy have been developed. Currently, various gene therapy strategies of malignant melanoma are being evaluated in multiple clinical trials carried out all over the world. They include gene modified cancer vaccines (GMTV) modified with genes encoding (i) cytokines or (ii) costimulatory molecules and dendritic cells modified with (iii) genes encoding tumor antigens or (iv) immunostimulatory factors. Since January 1996 in Department of Cancer Immunology USOMS, at GreatPoland Cancer Center in Poznan, Poland a GMTV has been tested in malignant melanoma patients. For the last 6 years more than 220 patients were enrolled into study of GMTV consisting of melanoma cells modified with genes encoding IL-6 and its agonistic soluble receptor (sIL-6R). More than 25% of objective clinical responses and significant life extension were observed. The encouraging results formed a basis for design of a phase III prospective, randomized clinical study. PMID- 12094638 TI - [Comparison of the classical and extended fronto-lateral laryngectomy considering the respiratory function of the larynx]. AB - In Szczecin during the period between 1973-1999, 107 patients were treated with classical fronto-lateral laryngectomy (LFLC) and 112 with extended fronto-lateral laryngectomy (LFLE) due to glottic cancer. We report 30 patients (27 male and 3 female)--13 after LFLC and 17 after LFLE (including 15 cases of LFLE with epiglottoplastic). 72 laryngovideoscopies, 87 spirometric examinations and 18 computerised tomographies were performed. Larynx lumen and 32 spirometric parameters were evaluated. Somewhat higher values of air flow parameters were state in patients qualified for LFLC than LFLE. Average value of the spirometric parameters were also a little bit higher after LFLC than after LFLE. Air flow through the larynx was sufficient to decannulate 11/13 patients after LFLC and 16/17 patients after LFLE. After both types of operations normal air flow was seen in 5 patients after LFLC and 2 patients after LFLE. Spirometric examination with flow-volume loop was the most useful in evaluation of respiratory function of the larynx. PMID- 12094639 TI - [Evaluation of ventilation parameters based on flow volume loop in patients with unilateral vocal cord paralysis]. AB - The vocal cord paralysis as a multidisciplinary problem requires commitment of different medical specialities. The diagnostic difficulties and necessity of laryngoscopic evaluation in cases suspected of vocal cord paralysis were pointed out. The analysis of ventilation parameters (flow-volume loop) was presented. The significant usefulness of inspiratory parameters, especially in upper airway obstruction was emphasized. The most useful parameters and coefficients were shown. Based on another authors data the criteria of airway obstruction classification were presented. The flow volume loop was found as a uniquely suited to the examination of upper airway obstruction. According to this examinations the unilateral vocal cord paralysis was defined as a variable extrathoracic obstruction which does not influence ventilation significantly. PMID- 12094640 TI - [Tracheopathia osteochondroplastica]. AB - The authors presents a very rare disease of trachea and bronchi on the basis of literature and their own case a 49-years old man. The diagnosis was made on the basis of CT trachea and bronchi, tracheobronchoscopy and biopsy with histopathological examination. The etiology of the disease is still unknown. It may be genetically determined. PMID- 12094641 TI - [Delayed treatment of laryngeal and hypopharynx cancer in the North-Eastern part of Poland in years 1986-1999]. AB - The authors estimated larynx and hypopharynx cancer in patients treated in the Department of Otolaryngology Medical Academy in Bialystok between 1996-1999 and compared the observed state to the data from 1986-1995. At the same time, we tried to determine the causes of delayed treatment based on patients' reports and medical case histories divided according to patients and medical procedures. As in years: 1986-1995, a significant percentage of tumour progression as also observed in the present day. The T3 and T4 tumour comprised about 65% of all larynx and hypopharynx cancers in particular five-year periods. The T1 and T2 tumours included only 34% of the cases. Clinical evaluation of the lymph nodes showed that high grade of tumour progression was accompanied by unilateral or bilateral lymph node metastases (N2 and N3--about 67%). Among the causes of the delayed treatment, patients' ignorance of the first symptoms (43-51%) and ignoring follow-up visit (30-32%) are being still the most dominant ones. According to medical procedures the delay ensued mostly because the effect of apparent improvement after the treatment (36%) and, unfortunately, due to wrong diagnosis (30-31%). Nowadays, it seems to be more difficult to obtain medical advice, specifically as far as specialists are concerned. PMID- 12094643 TI - [The evaluation of results of the surgical treatment in patients with otosclerosis]. AB - The authors presents hearing results in 216 patients with otosclerosis surgical treated. Averaged cochlear reserve in relation to the bone conduction determined before operation was used in this study. The follow-up was 2 months-10 years. Good results was proved in more than 93%, no change in 5% and deterioration in less than 2%. The cochlear reserve up to 10 dB obtained in 59% of all patients. The cases when no changes (and deterioration) after surgical treatment correlated to large anomaly evolution in the structure of the hole drum and extensive focus with otosclerosis. PMID- 12094644 TI - [Electrophysiological examinations (ABR and DPOAE) of hearing organ in hemodialysed patients suffering from chronic renal failure]. AB - Deterioration of function of hearing organ is one of the most important clinical problem in uremic patients with chronic renal failure. The present study aimed to assess the function of hearing organ using the brainstem auditory evoked responses (ABR), impedance audiometry and distortion product otoacoustic emission cochlear function (DPOAE) in 31 haemodialysed patients with chronic renal failure (16 females and 15 males, mean age 43.0 years). The control group consisted of 15 healthy subjects. The latency of the waves I, III, V and I-V interpeak in evoked response audiometry were significantly longer in the patients with CRF (chronic renal failure) compared to the control group. Measurement of DPOAE showed decrease of DPOAE level in patients suffering from CRF. A influence of single hemodialysis and treatment of hemodialysis by 6 months on ABR latencies and DPOAE values were not observed. PMID- 12094642 TI - [Labyrinthectomy with preservation of the cochlear duct and a reconstruction of the lateral wall in the vestibule]. AB - The authors described a technique and presented the results of the modified labyrinthectomy in 9 cases. A labyrinthectomy was performed during the removal of neuromas by translabirynthine approach, in advanced unilateral Meniere disease, in a huge perylimphatic fistula and in a case of labyrinthitis latens. In 3 cases of the neuromas removal the residual hearing was preserved. In remained cases of labyrinthectomy hearing was preserved partially. PMID- 12094645 TI - [Incidence of coexisting sensorineural hearing loss and secretory otitis media]. AB - There are currently no standard guidelines for assessing hearing in children who are evaluated for tympanostomy tubes. We describe the results of audiologic testing on 587 children, age 2 months to 17 years admitted to Pediatric Otolaryngology Department Bialystok for treatment of secretory otitis media. Ten children (1.7%) were found to have previously unrecognized sensorineural hearing loss. In four cases total unilateral deafness, in six others moderate to severe sensorineural bilateral hearing loss was diagnosed. Three other children referred to our clinic as sensorineural hearing loss were found to have secretory otitis media as the only or coexisting cause of deafness. Results of our study show the importance of age--appropriate hearing assessment as part of diagnostic procedure for secretory otitis media. PMID- 12094646 TI - [Social integration of children with multichannel cochlear implant in the mainstream education system]. AB - This paper is a preliminary report concerning result of social integration of children after cochlear implantation in the mainstream education system. It has been proofed that the presence of hearing impaired child in the class has an influence on other children. These other became more sensitive, less selfish and able to see needs of other human being. The tests were conducted on the group of 34 children and adolescents implanted in Poznan, learning in schools in the mainstream education system. PMID- 12094647 TI - [Selected marketing aspects in the hearing aids fitting and their distribution]. AB - The developing of free market in Poland gives medical firms of opportunities but also necessity of adoption for more and more individual needs of customers. This article presents some aspects of marketing on the hearing aids market. The conclusions are based on long observation and empirical research. Problems mentioned in it are connected with principles like distribution channels, specific customers services and promotion. It also shows relations between medical firms, specialists, doctors, audiologists and real customers. PMID- 12094648 TI - [Tinnitus in systemic diseases]. AB - Permanent or temporary degenerative changes in the internal ear causing tinnitus may occur with particular intensity in patients suffering from systemic disease (diabetes, hypertension, rheumatic diseases, kidney and thyroid gland diseases). Pathomechanisms of hearing impairment and the risk of tinnitus and its character in particular cases are discussed in the paper. The research was carried out on 1200 patients treated in the Laryngological Rehabilitation Centre in Poznan between 1.1.1998 and 04.2001 due to tinnitus. The analysis included the diagnosis of general health condition, general laryngological examination as well as additional tests. In the examined group 34% suffered from systemic diseases. Among them the highest percentage (47%) suffered from hypertension, 41% from hypercholesterolaemia, 22% from rheumatic diseases and 16% from diabetes. 96% of the patients had a long family history of diseases. The additional factor causing damage of the internal ear may be ototoxic drug used in the treatment of many systemic diseases. PMID- 12094649 TI - [Otoacoustic emissions and auditory brain stem responses in insulin dependent diabetic patients]. AB - The relationship between diabetes mellitus and hearing impairment has been studied for more than 100 years, and is still a matter of controversy. In our study hearing in diabetic patients was studied by tonal audiometry, brain stem auditory evoked responses and evoked otoacoustic emissions. The aim of this study was to evaluate the function of peripheral and central auditory pathway in subjects with insulin dependent diabetes mellitus (IDDM). Possible correlation between auditory function and microangiopathy (retinopathy or nephropathy) also have been investigated. Cochlear activity was evaluated by means of otoacoustic emissions. The functional changes in the brain stem were evaluated by ABR. OAEs and ABR were measured in 42 normally hearing IDDM patients aged between 21 and 42 years, and 33 aged- and sex-matched non-diabetic control subjects. IDDM patients were distributed into two groups, 17 patients without microangiopathy and 25 patients with microangiopathy. RESULTS: Both of the groups (diabetic and control) had normal and undifferentiated results in tonal and impedance audiometry. The mean amplitudes of various DPOAEs were significantly reduced in the diabetic group compared with control subjects. No correlations were found between microangiopathy and DPOAE amplitudes reduced. ABR latencies were longer in diabetic patients when compared with those of control subjects. These findings indicate a central disturbance in the auditory pathway. Microvascular complications (retinopathy or nephropathy) were associated only with the prolongation wave I latency. In conclusion, the combined use of different procedures for monitoring the central and peripheral portions of the auditory pathway in diabetic patients showed the existence of the alterations in the cochlear micromechanics and in the retrocochlear auditory pathway. PMID- 12094650 TI - [Failures, complications and treatment methods in otosclerosis operations]. PMID- 12094651 TI - [A comparison of water and air stimulated bithermal-caloric test and the usefulness of both methods in otologic surgery]. AB - The most commonly used method of testing the responses of the labyrinths is Water Stimulated Bithermal-Caloric Test of Fitzgerald-Hallpike. Before planning an otiatric surgery it is necessary to support by documentary evidence the responses of the vestibular organ in the ear planned to be operated on, however, irrigation with water may cause damage to structures of the ear and may intensify symptoms of the diseases and eventually make the treatment more difficult. In some centers where Water Stimulated Bithermal-Caloric Test were used alone, or a combination of air and water stimulated caloric test were performed, it seemed to be difficult to compare the results of both tests. In this study I tried to establish parameters of stimuli obtained in the Air Stimulated Bithermal-Calorics test that evoked nystagmus, similar to those obtained in the Fitzgerald-Hallpike water stimulated test. It was determined that the duration of nystagmus in both water and air stimulated bithermal-caloric tests were identical when the temperature of "cold" air wave was 26 degrees C, whereas "warm" air wave had the temperature of 48 degrees C and the time of stimulation for both tests was 80 seconds. PMID- 12094653 TI - [Evaluation of the ultrastructure of the vocal folds mucosa in patients with presbyphonia]. AB - Ultrastructure of the vocal folds mucosa was evaluated in 50 elderly patients. Study material included larynx specimens obtained from autopsy and postoperative material after the total laryngectomy due to the cancer of recessus piriformis with unchanged vocal folds. The ultrastructure of tunica mucosa was evaluated by means of the transmission electron microscopy with the use of Opton 900-PC microscope. In the control group the multilayer flat epithelium was found with the folded basal membrane, a large number of pericytes, scarce collagenous fibers in the stroma. The voice disturbances which occur during presbyphonia are conditioned by morphological changes in the epithelium, the basal membrane and the stroma of the vocal folds mucosa. Destruction of the epithelium cells with the enhanced vacuolar degeneration and enlarged intercellular spaces indicated oedemic character of presbyphonia. An increased number of collagenous fibers, vacuolar degeneration of fibroblasts with enlarged granular endoplasmic reticulum and an increased number of blood vessels in the stroma suggested an atrophic form of presbyphonia. PMID- 12094652 TI - [Esthesioneuroblastoma of nasal cavity and ethmoid sinus]. AB - This paper presents a rarely occurring tumor of nasal cavity and ethmoid sinus (esthesioneuroblastoma--2nd grade of the pathological grading system proposed by Hyams). It was observed in a woman aged 68. Moreover, there were described surgical therapy and repair of cerebral spinal fluid leaks using composite bone/mucosal graft of the lower turbinate as a donor material. The treatment was successful. PMID- 12094654 TI - [Rare case of Madelung disease]. AB - Madelung's disease, also known as benign symmetric lipomatosis (BSL), is a rare condition characterized by symmetric, painless fatty deposits arranged around the neck and shoulders girdle. This deformity is usually associated with chronic alcohol. The etiology remains unclear although some authors suggest that mitochondrial dysfunction represents the essential biochemical defect in BSL. This report describes a patient with this deformity. Lipomas were present in the pre-postauricular and nuchal area. PMID- 12094655 TI - [Rhabdomyosarcoma of parotid gland in 4 years old boy]. AB - The author described the case of parotid gland rhabdomyosarcoma in 4 years old boy. As there was no parents agreement to initial, radical surgical treatment, the patient underwent chemo- and radiotherapy, after which temporary remission of the disease was observed. The surgical treatment was performed for local recurrence, but despite radical parotidectomy, which was done, the progression of disease was occurred (with local recurrence and bone metastases). The patient died 3 years after initial diagnosis. PMID- 12094657 TI - [The case of gigantic pleomorphic adenoma of the parotid gland]. AB - The authors describe case of big size pleomorphic adenoma of the parotid gland. This tumor lead to cachexia and to elimination of life in society. Attention was paid on the surgical treatment of pleomorphic adenomas by superficial or total parotidectomies. Possibility of the post operative complications avoidance, even in such big cases was emphasized. Recurrence was not observed during 10 months follow up. Use of surgical treatment make that the patient was able to return to her previous job. PMID- 12094656 TI - [Cancers of nasopharynx in the material of ENT Department of Hospital in Kalisz]. AB - In the nasopharynx we can find malignant and non-malignant tumors. Authors present diagnostic difficulties on the base on literature and seven own cases. CT and NMR is needed to describe infiltration. Besides new methods radiotherapy and chemotherapy prognosis is poor. We have good results in early surgery in non malignant tumors. PMID- 12094658 TI - [Felix Anthony Erbrich (1874-1938)--a cofounder of Polish Otorhinolaryngology Society]. AB - The lived and works are presented of Feliks A. Erbrich (1874-1936), the well-know Warsaw otolaryngologist, a cofounder of Polish Otorhinolaryngological Society. He was a pupil of Alfred M. Sokolowski and Gustaw Killian. Dr Erbrich's achievements in the popularization of knowledge about pathology of tonsils and about broncho- and oesophagoscopy were strongly pointed out. The review of his scientific publications, practice and publishing activity were also described. Erbrich was a chairman of Polish Otorhinolaryngological Society and first chief of Clinic of Otolaryngology in Warsaw. PMID- 12094659 TI - [International otorhinolaryngology specialization programs]. PMID- 12094660 TI - [Analysis of sinusitis in patients of ENT ward from Interior Hospital in Olsztyn, in years 1989-1998]. PMID- 12094661 TI - [The report from the 2nd International Conference "A sound foundation through early amplification." Chicago, November 8-10, 2001]. PMID- 12094662 TI - Activating the community for nutritional improvement. AB - This presentation provides information on community-oriented action research conducted under the leadership of Dr. Rajammal P. Devadas, the recipient of the IUNS International Nutrition Award for 2001. Educating, activating, and energizing the community with empowerment have been the focus of the action oriented research. In the four decades of work presented, massive efforts have been made in the areas of infant and preschool child nutrition; nutritious noon meal programs for children; integration of nutrition, health, and sanitation concepts in the primary school curriculum; use of local foods to eradicate malnutrition; introduction of novel and underexploited foods; food-based approaches to overcome micronutrient malnutrition; equipping women for food and nutrition security; and nutrition education. These efforts have been activated through development of relevant food and educational materials, their introduction to the community, impact evaluation, follow-up, and implementation of the concepts in relevant national programs with massive training efforts. Many of the efforts outlined have formed the basis for regional and nationwide nutrition intervention strategies. The experiences gained and training efforts developed have gone beyond the country-level exposure to training and equipping nutrition workers in other countries. PMID- 12094663 TI - Control of vitamin A deficiency in Vietnam: achievements and future orientation. AB - Vitamin A deficiency is one of the major nutritional deficiencies in Vietnam. The first survey, conducted in 1985-1988 showed that the prevalence of severe xerophthalmia was seven times higher than the cutoff point established by the World Health Organization (WHO) to define vitamin A deficiency as a public health problem. The result of this survey strongly convinced the government to launch a program to control vitamin A deficiency, which started in 1988. The program strategies included nutrition education, universal distribution of high-dose vitamin A capsules to children aged 6 to 36 months in combination with national immunization days, and promotion of production and consumption of vitamin A-rich foods at the family level. The implementation network was set up based on the existing preventive health structure at all administrative levels. Organizations such as the women's union and other social sectors have participated actively in the program. Surveys conducted in 1994 and 1998 showed that the prevalence of clinical xerophthalmia was significantly lower than that identified in the baseline survey and below the WHO criteria for a public health problem. The achievements of our program have demonstrated that an effective vitamin A supplementation program can be implemented successfully by the preventive health network with active community participation. In the coming years, it will be important for our program to develop approaches other than vitamin A supplementation in order to maintain the past achievements. PMID- 12094664 TI - Empowering a tea-plantation community to improve its micronutrient health. AB - This project was designed to convince and empower management and plantation workers to improve their own nutritional health status and productivity. Plantations are generally bypassed by the government's primary health-care system. A nine-month intervention with iron (60 mg of elemental iron) and vitamin A supplementation and iodized salt was performed on the Balanoor Plantations in India. Of the women tea pickers, 99% (n = 334) received the supplements and bought the iodized salt from the plantation ration shop. Their mean hemoglobin level rose significantly from 11.0 to 11.9 g/dl. The women pickers gave the supplements to their families as well as themselves. The results were the same whether iron was given once or twice a week. The mean hemoglobin level of the women pickers rose significantly from 11.1 to 12.0 g/dl with the twice-weekly dose and from 10.9 to 11.8 g/dl with the weekly dose. The prevalence of clinical signs of vitamin A deficiency in the entire plantation population (about 2,500) was reduced significantly (from 19% to 4%), as was iodine deficiency (from 17% to 7%). Common health problems decreased from 88% to 54%. The number of patients referred to larger hospitals decreased significantly from 116 to 86. Absenteeism was not affected. PMID- 12094665 TI - The monetary value of human milk in Francophone west Africa: a PROFILES analysis for nutrition policy communication. AB - Using a simple and conservative methodology, we estimated the volume and monetary value of the human milk produced by lactating women in Francophone West Africa. In that region, children zero to 35.9 months old consume over 1.1 billion liters of human milk per year. However, suboptimal breastfeeding practices account for the loss of 175 million liters of human milk annually. If the human milk consumed by children zero to 35.9 months old were to be adequately replaced using commercial breastmilk substitutes, an annual expenditure of about 2 billion US dollars would be needed. At the household level, the annual replacement cost of human milk would amount to US$412 per infant. This is beyond the reach of most families in West Africa, where as many as 61% of families in some countries live on less than one US dollar per day. Appropriate policies to foster breastfeeding need to be developed and adequately implemented. Such policy action is more likely to occur if decision makers fully appreciate the monetary value of human milk. PMID- 12094666 TI - Targeting performance of three large-scale, nutrition-oriented social programs in Central America and Mexico. AB - This study evaluated whether three nutrition-oriented programs in Central America and Mexico have been successful in targeting those households most vulnerable to undernutrition and poverty. For each country, nationally representative data sets were used to estimate cutoff points dividing the population into 10 equal-sized groups according to child anthropometric measurements (age-standardized height) and household income (per capita household expenditures). Households meeting eligibility criteria were then assessed using special baseline surveys or national data obtained before implementation of the program. Children in these households were classified according to national deciles of height-for-age, and households were classified according to expenditure deciles. In spite of markedly differing targeting strategies, each of the programs was well targeted, with 45% (Honduras and Mexico) and 52% (Nicaragua) of children in eligible households coming from the lowest two deciles of the national distributions, and virtually none from the upper two deciles. Similar results pertained to household income. These experiences demonstrate that vulnerable households can be targeted relatively straightforwardly, and that the need to do this does not in every case imply household-level income screening. PMID- 12094667 TI - Dietary intake and nutritional status of young children in families practicing mixed home gardening in northeast Thailand. AB - One current initiative to assist rural Thai families to increase home food production and security is the implementation of home gardens that produce fish, small animals, and vegetables. This paper presents the results of an investigation comparing seasonal dietary intake and nutritional status among northeastern Thai children in mixed-gardening and nongardening families (n = 30 for each group). Assignment to the gardening group was based on the presence of a mixed garden, whereas nongardening subjects were randomly selected and matched for comparison. Statistical analysis (paired t-test) indicated that there was no significant difference in the observed biochemical variables (serum retinol, ferritin, hemoglobin) between groups at the p < or = .05 level. Nutritional status in regard to height-for-weight, weight-for-age, and weight-for-height Z scores was better among children of gardening families, although the differences were not significant. The small sample size and reported results indicate that the relationship between the practice of mixed home gardening and dietary intake and nutritional status needs further investigation. PMID- 12094668 TI - Evaluation of a rapid field tool for assessing household diet quality in Mozambique. AB - The Nutrition Section of the Mozambican Ministry of Health recently implemented a low-cost, rapid field tool for assessing the quality of household diets. Such tools can play an important part in targeting development assistance in countries where resources are scarce. This paper evaluates how well the tool performs at describing household dietary intakes in northern Mozambique and explores ways to improve it. The food-group classification and scoring system that form the core of the tool were applied to household data (n = 1,140) collected in a previous quantitative diet study in Nampula and Cabo Delgado Provinces. Using mean intakes as a criterion, the diet assessment tool performed well on all nutrients studied, except vitamin A. Those classified by the tool into the top group of diet quality had the highest mean intakes of energy, protein, and iron as well as the highest mean scores on the Mozambican Diet Quality Index, whereas those classified in the bottom group had the lowest intakes. Sensitivity rates for the diet assessment tool could be substantially improved by raising the cutoff point for an acceptable diet from the current threshold of 20 points to 23 points. Regression analysis was used to suggest other possible improvements. Such improvements were only marginal and do not justify field implementation, given the added complexity in classifying and scoring. This paper provides evidence that, with some minor changes, the Ministry of Health diet assessment method can be a useful tool in describing the dietary situation of groups of Mozambican households. Since this tool is both inexpensive and simple to use, there may be interest in adapting it for use in other low-income countries. A series of steps for doing so is outlined at the end of this paper. PMID- 12094669 TI - Effect of amylase-rich flour (ARF) treatment on the viscosity of fermented complementary foods. AB - Grains of cowpea and maize and slices of fresh cassava, cocoyam, plantain, and yam were steep-fermented in water, while flours from the same plant materials were fermented by backslopping for 24 to 30 hours. The pH and apparent viscosity of the gruels from the resulting flours were determined. Loss in weight due to fermentation was higher in fresh tubers than in dry grains. The pH of the flours decreased during fermentation. Measurements showed that the apparent viscosity only of gruels from flours produced by backslopping decreased after 24 hours. The apparent viscosity of gruels from steep-fermented flours was higher than that of the unfermented flours and those produced by backslopping. The apparent viscosity reduction of gruels from steep-fermented flours using amylase-rich flour (ARF) from five-day white sorghum malt was better with the prepared gruels than when applied to the flour-in-tap-water suspension before it was used to prepare gruels. Viscosity reduction using ARF was also better with the fermented gruels than with gruels from unfermented flours. The implications of these results for the formulation of complementary flour blends for infant feeding are discussed. PMID- 12094670 TI - Iodine stability in salt double-fortified with iron and iodine. AB - Deficiencies in small quantities of micronutrients, especially iodine and iron, severely affect more than a third of the world's population, resulting in serious public health consequences, especially for women and young children. Salt is an ideal carrier of micronutrients. The double fortification of salt with both iodine and iron is an attractive approach to the reduction of both anemia and iodine-deficiency disorders. Because iodine is unstable under the storage conditions found during the manufacturing, distribution, and sale of salt in most developing countries, the effects of packaging materials and environmental conditions on the stability of salt double-fortified with iron and iodine were investigated. Salt was double-fortified with potassium iodide or potassium iodate and with ferrous sulfate or ferrous fumarate. The effects of stabilizers on the stability of iodine and iron were followed by storing the salt under three conditions that represent the extremes of normal distribution and sale for salt in developing countries: room temperature (25 degrees C) with 50%-70% relative humidity, 40 degrees C with 60% relative humidity, and 40 degrees C with 100% relative humidity. The effects of stabilizers, such as sodium hexametaphosphate (SHMP), calcium carbonate, calcium silicate, and dextrose were investigated. None of the combinations of iron and iodine compounds was stable at elevated temperatures. Essentially all of the iodine was lost over a period of six months. SHMP effectively slowed down the iodine loss, whereas magnesium chloride, a typical hygroscopic impurity, greatly accelerated this process. Calcium carbonate did not have a sparing effect on iodine, despite contrary indications in the literature. Ferrous sulfate-fortified salts generally turned yellow and developed an unpleasant rusty flavor. Salt fortified with ferrous fumarate and potassium, iodide was reasonably stable and maintained its organoleptic properties, making it more likely to be acceptable to consumers. We confirmed that application of the iodine compounds as solutions resulted in a more even distribution of the iodine throughout the sample. The effect of the packaging materials was overshadowed by the other variables. None of the packaging materials was clearly better than any other. This may have been due to the fact that the polymer bags were not heat sealed, and thus some moisture penetration was possible. The results indicate that with careful control of processing, packaging, and storage conditions, a double-fortified salt could be stabilized for the six-month period required for distribution and consumption. Unfortunately, the processing and storage required are difficult to attain under typical conditions in developing countries. PMID- 12094671 TI - Weaning foods: a review of the Indian experience. AB - Although breastmilk is adequate to meet the energy and nutrient requirements of an infant up to four to six months of age, thereafter it is insufficient to sustain normal growth and needs to be supplemented with other foods, such as weaning foods. However, the capacity of a weaning diet to meet the protein and energy requirements of infants depends on its nutritional quality as well as its dietary bulk. This review highlights varieties of weaning foods, blend formulations, nutrient requirements, process characteristics, physicochemical parameters, quality evaluation, and other important aspects of developing a weaning food that satisfies all the requirements of the infant during the vulnerable transitional stage. Traditional weaning foods of India are briefly summarized. PMID- 12094672 TI - Seasonal undernutrition in rural Ethiopia: magnitude, correlates, and functional significance. PMID- 12094674 TI - Prevalence of bovine dermatophilosis in a tropical highland region of Ethiopia. AB - A study was made of bovine dermatophilosis on 540 animals at a high altitude (2400-2500 m above sea level) in Ethiopia. Eighty-two (15.4%) of the animals were affected by the disease which was more prevalent in local animals than was previously assumed. There was a significant (p < 0.05) difference in the prevalence among different age groups. However, the difference in the prevalence of the disease between males and females was not significant (p > 0.05). Associated risk factors included the relatively high average annual rainfall and humidity, tick infestation (76%) and management, related to the mixed crop livestock production system. The possible impact of the disease on Ethiopian agriculture is indicated. PMID- 12094673 TI - Comparative assessment of the erythrocyte osmotic fragility and of haematological and plasma biochemical values in the Nigerian white Fulani and N'dama breeds of cattle. AB - The blood profiles of the Nigerian White Fulani and N'dama breeds of cattle were compared. The White Fulani cattle had a significantly higher haemoglobin concentration (p < 0.001), mean corpuscular haemoglobin (p < 0.05), plasma sodium (p < 0.02), total protein (p < 0.01), albumin (p < 0.01) and globulin (p < 0.02), but lower neutrophil counts (p < 0.01) and osmotic fragility of erythrocytes than the N'dama cattle. The plasma potassium, calcium, bicarbonate, inorganic phosphate, albumin/globulin ratio, urea, creatinine, packed cell volume and mean corpuscular volume, and the erythrocyte, leukocyte, lymphocyte, eosinophil and monocyte counts were similar in the two breeds. PMID- 12094675 TI - Comparative pathogenesis of experimental infections with Salmonella gallinarum in local and commercial chickens. AB - The pathogenicity of a virulent strain of Salmonella gallinarum was studied in local chickens and commercial layers. Sixty 4-month-old chickens were used. Of these, 50 chickens were orally infected with S. gallinarum, comprising 25 commercial layers and 25 local chickens. Five chickens in each group were used as uninfected negative controls. The clinical signs and pathological features of acute, subacute and chronic fowl typhoid were observed in both groups. Chickens in both groups seroconverted, but the antibody titre was significantly higher (p < 0.001) in the commercial layers. The antibody titre remained high to the end of the experiment in all the surviving chickens. Only one commercial layer chicken died during the course of the experiment. The PCV decreased significantly (p < 0.001) in the infected chickens of both groups as compared to the controls. The viable cell count of S. gallinarum in the liver and spleen reached a maximum on day 9 after infection in both groups. However, there was a significantly higher cell count (p < 0.05) in the commercial layers. The severity of the disease appeared to be slightly greater in the commercial layers, although the susceptibility was similar in both groups. It was concluded that, under experimental conditions, local chickens are just as susceptible to S. gallinarum infection as are commercial layers. PMID- 12094677 TI - Ruminal fermentation kinetics in ewes offered a maize stover basal diet supplemented with cowpea hay, groundnut hay, cotton seed meal or maize meal. AB - The effect on ruminal digestive kinetics of supplementing maize stover (MS) with cowpea hay (CW), groundnut hay (GN), cotton seed meal (CSM) or maize meal (MM) was evaluated in a 5 x 5 Latin square using five Merino ewes with ruminal cannulae. CW and GN were offered at 30% w/w to MS, while CSM and MM were given at 15 g/kg0.75 per day. There were significant time (p < 0.001) and time x diet (p < 0.05) effects on pH but the effect of diet alone was not significant (p > 0.05). There was a significant (p < 0.05) increase in the concentration of ammonia with time on the supplemented diets. There were no significant (p > 0.05) differences in the concentrations of total volatile fatty acids (VFAs). Butyrate increased with supplementation (p < 0.05). Supplementation increased the concentrations of branched-chain and total VFAs with time (p < 0.05). The non-glucogenic ratio was not significantly (p > 0.05) lower on supplemented diets owing to the higher butyrate concentration. The degradation curve constants were not significantly different (p > 0.05), the average effective dry matter degradation being 27.80% +/- 0.618% at 0.05 h-1 flow rate. The chemical and physical characteristics of the cell wall of MS may have presented few sites for microbial colonization, so limiting degradation. These results showed that small quantities of forage supplements can improve the ruminal environment in animals subsisting on low quality forages and form a basis for comparison with bought-in protein and energy sources. PMID- 12094676 TI - A comparison of the prevalence and burdens of helminth infections in growers and adult free-range chickens. AB - Matched samples of 100 chickens of each of growers and adult rural free-range chickens in Morogoro, Tanzania, were purchased from the beginning to the end of the long rainy season. At necropsy, the trachea, the gastrointestinal tract and the oviduct were examined for helminth infections. The helminth species isolated comprised 18 nematodes and 8 cestodes but no trematodes. Tetrameres fissispina is a new record in Tanzania. All the chickens harboured at least three different helminth species. Growers contained 4-14 and adults 3-12 helminth species. The number of species isolated per chicken increased as the rainy season advanced. The prevalence of the following species were significantly higher in growers than in adults (p < 0.05); Ascaridia galli (69% of growers, 29% of adults); Syngamus trachea (14%, 3%); Tetrameres americana (94%, 82%); Trichostrongylus tenuis (43%, 7%); Choanotaenia infundibulum (15%, 6%); Davainea proglottina (9%, 2%); and Raillietina tetragona (36%, 21%). Allodapa suctoria (3%, 20%) and Capillaria annulata (1%, 10%) had a significantly lower prevalence in growers than in adults (p < 0.05). There were significantly higher worm burdens (p < 0.05) in growers than in adults for A. galli, Capillaria caudinflata, R. tetragona, S. trachea, T. americana, T. fissispina and T. tenuis. Conversely, A. suctoria and C. annulata showed significantly higher worm burdens in adults (p < 0.05). The sex of the chickens influenced the burdens of Heterakis brevispiculum (p < 0.05). There was an interaction effect such that growing males and adult females had statistically higher (p < 0.05) burdens of T. tenuis and A. suctoria, respectively. PMID- 12094678 TI - Assessment of molasses-urea blocks for goat and sheep production in the Sultanate of Oman: intake and growth studies. AB - The goal of this study was to develop a molasses-urea block (MUB) for purposes of supplementing trace minerals to domestic ruminant livestock in Oman. To accomplish this, the utility of molasses and date syrup as fermentable energy sources, of straw, date flakes and wheat bran as fibre sources, and of cement and lime as binders were evaluated. The proportion of cement needed for adequate hardening of the block was also studied. Molasses- and date syrup-based blocks hardened equally well. However, the higher cost of date syrup precluded its use. Wheat straw yielded a low-density block that hardened slowly. Date fibre retained moisture and hardened extremely slowly. Wheat bran-based blocks hardened quickly and yielded dense blocks. Hence, wheat bran was judged to be the superior source of fibre. Lime did not effectively bind the blocks. A cement content of 15% allowed hardening of the blocks within 2-3 weeks. A level of 10% cement in the block reduced the hardening rate by about 50%. Sheep and goats consumed both the straw- and wheat bran-based blocks but at different rates. Consumption of the straw-based block by sheep ranged from 50 to 179 g/head per day, whereas the denser wheat bran-based block was consumed at a rate of 8-20 g/head per day. Consumption of the straw-based block by goats was low (8 g/head per day) compared to that of wheat bran-based blocks (16-24 g/head per day). On the basis of the intake of the bran-based block by sheep, a block was designed that would provide approximately 50% of an animal's trace mineral requirements per day. This block consisted of 45% molasses, 10% urea, 5% trace minerals, 2.5% NaCl, 22.5% wheat bran and 15% cement. Sheep consuming this block gained more weight than sheep fed a conventional mineral block or sheep receiving no mineral supplementation. MUBs are inexpensive (9.5 US cents/kg). We conclude that MUBs have utility for providing trace elements in ruminant diets. PMID- 12094679 TI - Studies on the use of Colocasia esculenta (taro cocoyam) in the diets of weaned pigs. AB - Sixty weanling Large White x Duroc pigs were allocated to five groups of 12 pigs per group and fed on one of five diets. The five diets comprised 0, 50% and 100% unboiled, sun-dried taro cocoyam cormels (Colocasia esculenta) and 50% and 100% boiled, sun-dried taro cocoyam cormels as replacements for maize. The levels of some antinutritional factors were also determined in both boiled and unboiled, sun-dried taro cocoyam. Boiling reduced (p < 0.05) the amounts of the antinutritional factors in the taro cocoyam cormels. There were no significant differences (p > 0.05) in feed intake, weight gain or feed efficiency between the diets containing boiled taro cocoyam cormels. However, for unboiled, sun-dried taro cocoyam cormels, there were significant differences (p < 0.05) in weight gain and feed efficiency, these being depressed at more than 50% replacement of maize. This may be due to the relatively high amounts of antinutritional factors in the unboiled, sun-dried taro cormels. Boiled taro cocoyam cormels were comparable to maize as an energy source in the diets of weanling pigs. PMID- 12094680 TI - The effect of mixing and translocating juvenile ostriches (Struthio camelus) in Botswana on the heterophil to lymphocyte ratio. AB - The possibility was investigated that translocation of juvenile ostriches from concrete-paved to sandfloored pens and mixing of batches of ostriches after such translocation constitute a stress strong enough to evoke changes in the ratio of heterophils to lymphocytes. Blood smears were obtained from 15 ostriches out of a group of 25 birds 4 and 2 days before and then 2 and 4 days after translocation. The heterophil to lymphocyte ratio changed from 0.27 and 0.37 on days 4 and 2, respectively, before mixing and translocation to 0.53 and 0.84 on days 2 and 4, respectively, after translocation. Mixing and translocating juvenile ostriches appears to constitute stress. This information is important for the onfarm management of juvenile ostriches to enhance their welfare and productivity. PMID- 12094681 TI - Performance of broilers fed on diets containing different amounts of chaya (Cnidoscolus aconitifolius) leaf meal. AB - The performance and gut measurements of broilers fed on diets containing different amounts of chaya (Cnidoscolus aconitifolius) leaf meal (CLM) were examined in two experiments. In the first experiment, 60 Hubbard chickens (30 males and 30 females; 2 weeks old) were fed on five maize diets; these were formulated using 0, 150 (CLM150), 250 (CLM250) or 350 (CLM350) g CLM/kg, and the fifth diet contained soyabean. In the second experiment, 148 Ross male chicks, 1 day old, were fed on three isonitrogenous and isoenergetic maize-soyabean-based diets, which included 0 (control), 150 (C150) or 250 (C250) g CLM/kg. The diets were offered ad libitum for 2 or 3 weeks in the first and second experiments, respectively. Food intake, weight gain and the food:weight gain ratio were recorded. The weight of the gizzard and intestine and the weight and length of the caeca were also determined in the second experiment. In experiment 1, the birds fed on the maize-soyabean diet had a higher (p < 0.05) weight gain and final weight than birds fed on maize only or on the CLM150 diets. There were no differences for any of the variables studied between the birds fed on the maize soyabean diet and those fed on the CLM250, nor between males and females. In the second experiment, weight gain, food intake and the food:weight gain ratio for birds fed on C250 were lower (p < 0.05) than those in birds fed on either the control or C150 diets. The weights of the gizzard and intestine were the lowest and the highest, respectively, in birds fed on C250 (p < 0.05). The length and weight of the caecum from birds fed on the control diet were lower (p < 0.05) than those of birds fed on either the C150 or C250 diets. The results from this study suggest that CLM may be included up to 150 g/kg in commercial diets without having an adverse effect on poultry performance, and may also be mixed with maize up to 250 g/kg to improve the performance of chickens fed on low-protein diets. PMID- 12094683 TI - [Stereotactic gamma radiosurgery]. PMID- 12094684 TI - [Stereotactic irradiation using linear accelerator]. PMID- 12094685 TI - [Quantitative analysis of plasma concentration of barbiturate for diagnosis of brain death]. AB - Organ transplantation from brain death patients started in Japan in 1997. However it is difficult to diagnose brain death in patients treated with barbiturate therapy. In this study, the influence of long continuous administration of barbiturate on diagnosis of brain death was investigated by measuring plasma concentration of barbiturate. In 15 patients treated with barbiturate therapy, plasma concentrations of thiamylal were measured by liquid chromatographic apparatus every day until it's level decreased below 0.1 microgram/ml after cessation of continuous administration. At the same time, plasma thiamylal levels were checked on the day when burst-suppression (b-s) pattern had disappeared in 9 cases, light reflex of pupil appeared in 7 cases and spontaneous respiration had been detected by trigger lamp in 11 cases. The plasma concentrations of thiamylal on the day when b-s pattern had disappeared differed clearly among the cases in the range of 8.8 to 37.9 micrograms/ml. Those cases in which light reflex of the pupil had been recognized were also different in the range of 17.8 to 57.8 micrograms/ml. The cases in which spontaneous respiration had been detected were in the range of 4.4 to 23.0 micrograms/ml. These concentrations varied about 4, 3 and 5 times among the cases examined. The intervals between cessation of continuous administration of thiamylal and the decrease of plasma concentration to below 0.1 microgram/ml also varied from 2 to 14 days from case to case. The minimum concentration of thiamylal on the day when b-s pattern had disappeared, light reflex of the pupil had been recognized and spontaneous respiration had been detected was 8.8, 17.8 and 4.4 micrograms/ml respectively. These results suggest that diagnosis of brain death in patients treated with barbiturate therapy is able to be made when the plasma thiamylal level is below 4.4 micrograms/ml. PMID- 12094686 TI - [The development of three dimensional video microscope system for clinical application in neurosurgery]. AB - OBJECTIVE: The employment of surgical microscopy in the field of neurosurgery has significantly contributed not only to the advancement of surgical technique, but also the operative outcome as well. This technology has allowed surgeons to meet the demands for increasingly difficult levels of surgery, which usually require a long operative time. However, the surgeon and the assistant are restricted in their posture for a long time, leading to excessive muscular and ocular stress. To solve these problems, we have developed and clinically applied a three dimensional video microscope system for surgery, using a two binocular camera method, that allows surgical procedures to be performed over a video monitor. METHOD: Two 3CCD cameras were installed on the assistant's lens attachment and the camera lens attachment of the microscope for neurosurgery, manufactured by Carl Zeiss Co., Ltd. The surgery was performed by viewing the three dimensional picture on the monitor, which was created by processing the images from the cameras by a time-division method. In November 1998, the prototype was completed and, with numerous modifications, its clinical application has become possible. Currently, we have used it in 42 clinical cases. RESULTS: 1) Instead of viewing the surgical field through a conventional surgical microscope, the surgeon and the assistant were able to perform microsurgery by viewing the stereoscopic vision on the video monitor, through polarized glasses. 2) The assistant is provided with a separate video monitor, so that the assistant can freely adjust the angle of vision 90-180 degrees, according to where he stands. 3) The nurses, resident surgeons and students can view the same three dimensional images displayed to the surgeon. This effectively contributes to the educational environment. CONCLUSION: Even during a long surgical procedure, the posture of surgeon and assistant became less restricted since there was no need to look through the eyepiece of the microscope and led to significant decrease in physical fatigue and ocular strain. In the 42 clinical trials, the operation proceeded uneventfully, without any serious complications. PMID- 12094687 TI - [Rupture of previously documented asymptomatic unruptured aneurysms--aneurysm size: risk factor for aneurysm rupture]. AB - The natural history of asymptomatic unruptured aneurysms is not clear. We conducted a follow up study of 100 patients (since 1993) with 122 asymptomatic unruptured aneurysms that had not been operated on. We report five patients with previously documented asymptomatic unruptured aneurysms smaller than 10 mm in diameter that subsequently ruptured. Among the 100 patients, five had suffered subarachnoid hemorrhage (SAH) due to rupturing of an aneurysm. Of the 5 cases, 1 was male and 4 were female, with ages ranging from 59-73 years (mean age, 68 years). The aneurysms were on the MCA in 3, on the BA-SCA in 1, on the IC-PC in 1. The maximal diameter of the aneurysms at diagnosis ranged from 4.5 to 8 mm. The period from discovery to SAH was from 4 to 69 months and the cumulative rate of rupture of the aneurysms was 1.5 percent per year. Four of the 5 cases increased in size after the rupture. In our series, 2 of the 5 cases showed enlargement and the development of an aneurysmal bleb in the follow up MRA and 3D CTA. The present study demonstrates that five asymptomatic unruptured aneurysms less than 10 mm in diameter subsequently ruptured. We ought to seriously consider the assertion published in the New England Journal of Medicine (Dec. 10, 1998), that unruptured aneurysms less than 10 mm in diameter have a very low probability of subsequent rupture. PMID- 12094688 TI - [111In-DTPA cisterno SPECT for the diagnosis of CSF rhinorrhea: case report]. AB - The authors report on a patient with postoperative CSF (cerebrospinal fluid) leakage subsequent to transsphenoidal surgery where cisterno SPECT (single photon emission computed tomography) demonstrated the precise location of a CSF fistula. Seven months after surgery, the patient suffered from CSF rhinorrhea and headache. MRI (Magnetic Resonance Imaging) revealed significant contrast on T1 weighted images resulting from measurements in the right sphenoid sinus, which were hyperintense relative to CSF. On the basis of signal intensity differences, MRI could not distinguish between CSF leakage and postoperative scarring. Therefore, we performed cisterno SPECT at the same time as RI cisternography with intrathecal lumbar injection of 111In-DTPA which revealed dramatic accumulation of the tracer in the right sphenoid sinus. The patient underwent re-operation via a transsphenoidal approach, and the CSF leakage was repaired using fat-in-fibrin glue and the sella floor was reconstructed by hydroxyapatite platinge. These results suggest that cisterno SPECT may be useful in identifying the precise location of CSF fistulae, while other techniques fail to show evidence of CSF leakage. PMID- 12094689 TI - [Unilateral exophthalmos caused by a prolactin producing ectopic pituitary adenoma: case report]. AB - OBJECTIVE: We report a case of a patient with a prolactin (PRL) producing ectopic pituitary adenoma presenting a unilateral exophthalmos. CASE: This 70-year-old woman presented an ophthalmologist with progressive left-sided exophthalmos over the previous 2 months. Bone window CT scan revealed extensive bony destructions of the skull base including the clivus, sphenoid sinus and medial aspect of the middle cranial fossa. Gd-DTPA MRI revealed an abnormal enhancement lesion in the sphenoid sinus, but no abnormal enhancement was seen in the sella turcica. Since these findings suggested malignant tumors of the cranial base, several biopsies through the transnasal route were carried out to confirm the diagnosis. This procedure caused the complication of cerebrospinal fluid (CSF) leakage. Because the biopsy specimen revealed a PRL producing adenoma (serum PRL-level 645.7 ng/ml), the patient was admitted to our department. On admission neurological examination showed an exophthalmos with external ocular movement disorders and disturbance of visual acuity on the left side. She underwent transsphenoidal surgery to remove the tumor and to reconstruct the sphenoid sinus and the sellar floor. Surgical exploration revealed a yellowish and soft tumor underneath the normal mucous membrane in the sphenoid sinus. The sellar floor was destructed extensively, but the dura mater of the pituitary fossa was intact except for a small pin-hole which was thought to be produced during the several biopsy procedures. No surgical procedure was applied to the intrasellar region. The sphenoid sinus was packed with a piece of fascia and fat applied with the aid of fibrin glue to prevent CSF leakage. RESULT: The patient followed a satisfactory postoperative course. Her visual acuity disturbance and exophthalmos disappeared one year after surgery. Postoperative serum PRL level remained high (66.9 ng/ml), but, subsequently, was normalized (9.5 ng/ml) with a bromocriptine therapy (15 mg daily). CONCLUSION: As far as we are aware, this is the first case report of an ectopic pituitary adenoma causing unilateral exophthalmos. Although it is extremely rare, pituitary adenomas should be kept in mind in a differential diagnosis of exophthalmos. PMID- 12094690 TI - [Successful treatment of a huge meningeal hemangiopericytoma using Preoperative Autologous Transfusion and Hemodilutional Autologous Transfusion: case report]. AB - We report successful operations for a meningeal hemangiopericytoma using sufficient amounts of Preoperative Autologous Transfusion (PAT) and Hemodilutional Autologous Transfusion (HAT). A 23-year-old woman with amenorrhea and bilateral visual field disturbance was found to have a huge intracranial tumor. MRI showed a well-enhanced cystic mass in the left middle fossa, suprasellar, intrasellar, sphenoidal sinus, and cavernous sinus. Preoperatively, the tumor was thought to be a cystic pituitary tumor or meningioma. Surgical removal was planned in three steps. The first operation was carried out via the transsphenoidal approach. Total blood loss was 1348 ml and 2 MAP infusion were required to control bleeding. Histopathological diagnosis was hemangiopericytoma. After preparation of PAT 400 ml and HAT 800 ml, we carried out the second partial removal operation mainly via the interhemispheric approach. Total blood loss was 1829 ml and required autologous transfusion only. After preparation of PAT 1200 ml and HAT 400 ml, the last total removal operation was carried out mainly via the pterional and subtemporal approach. Total blood loss was 1813 ml and required autologous transfusion only. We needed 2 MAP infusion in the first operation, but were able to perform total removal successfully without homologous blood transfusion because a sufficient amount of PAT and HAT had been prepared preoperatively. Hemangiopericytoma required postoperative radiation therapy to avoid local recurrence. After successful removal of the tumor surgically, postoperative radiation therapy was able to be carried out efficiently. PMID- 12094691 TI - [Frontal atypical teratoid/rhabdoid tumor evaluated by single-photon emission computerized tomography and positron emission tomography]. AB - A case of frontal atypical teratoid/rhabdoid tumor (AT/RT) was presented in the findings of single-photon emission computerized tomography (SPECT) using 201Tl chloride (Tl) and 123I-metaiodobenzylguanidine (MIBG), and positron emission tomography using 18F-fluorodeoxyglucose (FDG) and (11C-methyl)-L-methionine (Met). A 16-year-old female had an episode of Jacksonian seizures one month prior to admission. CT scans showed an isodensity mass with heterogeneous enhancement in the left frontal lobe. MR images revealed a tumor with solid and cystic components and perifocal edema. SPECT demonstrated intense accumulation of Tl and MIBG in the enhancing lesion 15 min and 30 min after intravenous injection of tracers, respectively. The 6-hr delayed SPECT showed no retention of MIBG in the enhancing lesion. FDG-PET and Met-PET revealed high uptake of tracers in the enhancing lesion. Met did not accumulate in the frontal white matter, which appeared hyperintense on T2-weighted MR images. The patient underwent an uneventful extirpation of the solid mass, where Met had accumulated. Microscopically, the solid tumor contained rhabdoid cells, spindle-shaped cells resembling mesenchymal cells, and nests of small cells. The tumor cells were immunoreactive for vimentin, cytokeratin, and epithelial membrane antigen. The MIB-1 labeling index was 25%. The histological diagnosis was AT/RT. Postoperative course was uneventful. A dose of 32.4 Gy was administrated to the whole brain and a boost of 27.8 Gy to the T2-hyperintensity lesion. Five months after the radiotherapy, MRI showed neither abnormal enhancing lesions nor the T2 hyperintensity lesion. Multifarious studies using SPECT and PET are useful for differential diagnosis and for choosing optimal therapeutic strategy for this type of tumor. PMID- 12094692 TI - [An intradiploic epidermoid cyst of the skull in infancy: case report]. AB - An intradiploic epidermoid cyst of the skull in infancy is rare. We report a case of a 7-month-old girl with an intradiploic epidermoid cyst of the left parietal bone. The patient was admitted to the department of pediatrics in, our hospital in March, 2000, complaining of a lump in the scalp with a diameter of 1.5 cm. The pediatrician doubted that it was a case of Langerhans cell histiocytosis (LCH). In addition, as a result of further tests there were no systemic findings to suggest LCH. The skull x-ray showed round radiolucency of the left parietal bone. CT scans showed an iso density intradiploic mass with destruction of the outer table. Because of the fact that the mass was enlarging, the patient was admitted to our neurosurgical service in April, 2000. We excised the lesion to confirm the histological findings. The histological diagnosis was epidermoid cyst. We discuss the clinical feature and treatment strategy for intradiploic epidermoid cyst in infancy. PMID- 12094693 TI - [Controversy and reform proposals concerning insured medical care in neurosurgery]. PMID- 12094694 TI - [Molecular diagnosis of cancer--the progress of its applications for diagnosis and treatment of cancer]. PMID- 12094695 TI - [Molecular alterations in esophageal cancer]. AB - The clinicopathological characteristics of esophageal cancer have gradually been clarified using molecular biologic methods developed over the past 20 years. For example, amplification of the c-erb B gene is a prognostic factor and predictive of lymph node involvement, while the amplification of the cyclin D1 gene is also a prognostic factor and predictive of distant organ metastasis. Alteration of the p16 gene is also a prognostic factor and predicts lymph node involvement. As telomerase activity is almost a unique phenomenon of cancer cells, highly sensitive detection of esophageal cancer cells in the peripheral blood can be performed. Recently, such new methods as comparative genomic hybridization analysis and cDNA microarray analysis have been used to determine meaningful genetic changes. For therapeutic purposes, although tailor-made therapy has been proposed for several years, the validity of these approaches should be confirmed in a well-designed clinical trial. As molecular targeted therapies, tyrosine kinase inhibitors of epidermal growth factor receptor (EGFR) and monoclonal antibodies against EGFR are being studied in clinical trials in Western countries. A clinical trial of p53 gene therapy against esophageal cancer is also promising. PMID- 12094696 TI - [Molecular diagnosis of gastric cancer]. AB - Based on the genetic and epigenetic abnormalities of cancer-related genes during the development and progression of gastric cancer, we have established a system of molecular-pathological diagnosis as a routine service in Hiroshima. Approximately 5,000 lesions of the stomach have been diagnosed and useful information on differential diagnosis, grade of malignancy, and tumor multiplicity has been obtained using this system. Further research on systemic gene expression profiles, epigenetic alterations, and genetic polymorphisms will improve the quality and efficiency of diagnosis. With the application of novel knowledge on molecular carcinogenesis and microarray techniques, molecular diagnosis will identify the characteristics of individual cancers and persons, which will lead to the development of personalized medicine. PMID- 12094697 TI - [Recent advances in the diagnosis and treatment of colorectal cancers]. AB - We review recent developments in the diagnosis and treatment of colorectal cancer. In diagnosis, the detection of the APC gene mutation in stool samples has had a strong impact because of its practical usage for mass screening. Several reports have indicated that the molecular detection of a very small number of cancer cells in the peripheral blood, bone marrow, or lymph nodes yields adequate information on recurrent disease. Several new markers were reported as a significant indicators of cancer development, invasion, or metastasis. In treatment, determination of microsatellite instability may be useful to select good candidates for 5-fluorouracil-based chemotherapy. New molecular targets have been reported, including epidermal growth factor receptor inhibitor, matrix metalloproteinase inhibitors, etc. Gene therapy with p53 or FHIT had undergone successful clinical trials. Tumor-specific immunotherapy with dendritic cell vaccination has also been achieved. In conclusion, many efforts to overcome colorectal cancer will soon open new therapeutic windows. PMID- 12094698 TI - [Genetic detection and clinical applications in patients with hepatocellular carcinoma]. AB - This paper provides a review of the known genetic diagnostic indicators of liver cancer. The correlation between the genetic diagnosis and clinical outcome in patients with hepatocellular carcinoma (HCC) has been widely reported, based on the detection of liver-specific mRNA or tumor DNA in the blood. Our results suggest that an alteration of alpha-fetoprotein (AFP) mRNA in peripheral blood from HCC patients during the perioperative period might permit the prediction of early recurrence after surgery. The presence of circulating HCC cells may be indicative of metastasis if liver-specific AFP mRNA is detected in the peripheral blood. However, some studies showed that sensitive RT-PCR might possibly give rise to false positivity because nontumor hepatocytes would also express low levels of AFP mRNA. Recently, quantification of AFP mRNA for HCC cell detection using real-time PCR or semiquantitative RT-PCR has proven useful in the prediction of metastasis/recurrence. On the other hand, circulating liver tumor DNA such as p16 and p15 methylation and mitochondrial mutations in the plasma and serum of liver cancer patients might be useful for monitoring, similar to the tumor markers. In future, HCC-specific genes and genes sensitive to radiation and chemotherapy are expected to have wide-spread clinical applications. PMID- 12094699 TI - [Genetics of pancreatic cancer: recent advances in molecular diagnosis]. AB - Pancreatic cancer is an important cause of death from cancer throughout the world. Until recently, pancreatic cancer was a poorly understood disease. Research in the past decade has shown conclusively, however, that pancreatic cancer is primarily genetic in nature. Inactivation with a variety of tumor suppressor genes such as p53, p16, and DPC4, and genome-maintenance genes such as BRCA2, coupled with the activation of oncogenes such as K-ras, are a few of the mutations that trigger the growth of cancerous cells. The genetic profile of pancreatic cancer has reshaped the nomenclature describing histological progression in pancreatic ductal tumorigenesis. K-ras mutations frequently occur early, whereas changes in the expression and genetic integrity of the p16 gene appear in intermediate lesions, and the inactivation of the p53 and DPC4 genes and activation of telomerase occur late in the neoplastic progression. Although the majority of pancreatic cancers occur sporadically, a minority has been shown to aggregate in families and has aided our understanding of pancreatic tumorigenesis. An improved understanding of the genetics of pancreatic cancer should lead to the development of gene-based screening tests and novel rational therapies. PMID- 12094700 TI - [Genetic investigations for lung carcinoma]. AB - Lung carcinoma is a common malignant disease in adults. Various genetic changes associated with small cell and non-small cell lung carcinoma are now known. There are two types of genetic change which influence tumor growth and patient prognosis. Among oncogenes, c-myc, K-ras, and erbB-2 are considered to play important roles in lung carcinogenesis. The p53 suppressor gene is highly expressed in small and non-small cell carcinoma. To differentiate between double primary lung carcinomas and intrapulmonary metastasis, or between primary lung carcinoma and metastatic lung tumor, the analysis of gene mutations, such as of p53 and K-ras, appears to be very useful. PMID- 12094701 TI - [Molecular diagnosis of breast cancer]. AB - Alterations in protooncogenes and tumor-suppressor genes at the DNA and/or protein level indicate the biological properties of individual breast cancers. Information on these genetic and protein alterations is considered to be useful diagnostically and these alterations are now being applied in routine diagnosis or are under investigation for diagnostic application in the areas of: 1) determination of indications for specific therapies for advanced or metastatic breast cancers; 2) prediction of the response of primary tumors to adjuvant or neoadjuvant systemic therapies; and 3) identification of patient groups at high or low risk of recurrence from node-negative breast cancer. PMID- 12094703 TI - [The reinstatement criteria and follow-up system after returning to work]. AB - It is possible to think that some consideration must be given to various aspects for those returning to work after having taken temporary leave to receive treatment for stress-related disorders or psychiatric disorders. In this study, the policy aimed at solving the problems was examined by pointing out its strengths and weaknesses of the reinstatement criteria and follow-up system after returning to work at M electrical machinery manufacturing group. For the workers to maintain good social adequacy and tertiary preservation it is necessary for the work place to provide a reinstatement system with an attitude of acceptance. Therefore, it is reconfirmed that the most important thing, as a concrete action, is to give good care after reinstatement. Furthermore, it is essential that efforts be made to remove misconceptions or prejudice in society and among workers by not merely managing the system as a mechanical businesslike procedure, but also enlightening people about the meaning of the system. PMID- 12094702 TI - [Thyroid carcinoma: genetics, diagnosis, clinical features, and surgical treatment]. AB - The genes implicated in thyroid carcinoma can be categorized as oncogenes or tumor-suppressor genes. The RET oncogene has well-established roles in the development of both medullary and papillary thyroid carcinoma (PTC). Genetic testing for the germline RET mutation is commonly performed, and prophylactic thyroidectomy is carried out at an early stage. The demonstration of a RET rearrangement in a PTC patient may be prognostic factor. TSH-R and Gs alpha are associated with the development of toxic thyroid adenoma (AFTN). The ras oncogene is implicated in the early stages of development of several tumor types. In conclusion, germline screening for RET mutations is now commonly undertaken in patients with medullary thyroid carcinoma. PMID- 12094704 TI - [A proposed method for evaluating exposure conditions from measured values when sampling duration less than 8 hours]. AB - In order to evaluate occupational exposure conditions, it is necessary to determine the 8-hr time-weighted average (8-hr TWA) of exposure concentrations and compare the 8-hr TWA value with the permissible exposure limit, but 8-hr sampling is difficult for the outside laboratories because of trip time. For working environment measurement, because sampling duration in a unit work area may be one hour, the measurement can be conducted in several work areas in a day. If exposure conditions can be evaluated with the 1-hr TWA values, sampling to determine the exposure concentration would be efficient and able to be completed during the work area sampling. From this point of view, methods for evaluating exposure conditions with 1-hr or more TWA values are proposed in this paper. PMID- 12094705 TI - [How many ergonomics interventions do we have in the field of occupational stress?]. AB - This paper describes handling stress in the workplace from the viewpoint of work conditions and ergonomics. The term expressed in Japanese as "sagyokanri" corresponds to the term "work conditions and ergonomics" in English, which is used internationally in the field. Therefore, "sagyokanri" means considering how to handle work and stress by using ergonomics. The Japanese Journal of Ergonomics is quoted in this discussion of the definition of stress and the lack of countermeasures taken with respect to work conditions and ergonomics as they relate to stress. Finally, it touches on the SACL (Stress Arousal Checklist), one method of evaluating stress which is not very familiar to those in the occupational health field in Japan. PMID- 12094706 TI - [Improvement of work environment]. AB - As a result of a literature review on methodology and the effectiveness of work environment oriented workplace stress reduction, it is suggested that improvement of the work environment is effective in reducing workers' job stressors and psychological and physiological stress reaction, from a limited number of controlled studies including RCT, as well as a number of case studies and before after comparison studies. An expert-guided, supervisor and worker participated approach also seems effective. The author proposes five practical steps toward an effective improvement of the work environment based on the review. Remaining topics include the need for more well-designed studies, investigation of the relationship between intervention type and outcome, as well as modifying effects according to social situations, and instruments for implementation. PMID- 12094708 TI - [Clinical effects of allylestrenol on patients with benign prostatic hyperplasia (BPH) evaluated with criteria for treatment efficacy in BPH]. AB - One hundred and twenty-nine patients with benign prostatic hypertrophy (BPH) were registered and treated with allylestrenol. Allylestrenol was administered at a dose of 50 mg/day given twice a day for 16 weeks. Out of 129 patients with a mean age of 67.8 years old, 92 cases completed the study and 48 cases with moderate symptoms were objectively evaluated with "Criteria for Treatment Efficacy in BPH" proposed by The Japanese Urological Association in 1997. Prostate volume was significantly decreased from 32.7 +/- 11.9 to 27.4 +/- 11.2 ml (mean +/- SD), and maximum flow rate was significantly increased from 8.4 +/- 3.4 to 10.8 +/- 5.0 ml/sec. Residual urine volume was significantly decreased from 62.4 +/- 57.4 to 37.0 +/- 38.7 ml. IPSS was significantly decreased from 15.3 +/- 4.9 to 9.9 +/- 4.0, and QOL index was significantly decreased from 4.4 +/- 0.8 to 2.7 +/- 1.2. The efficacy of allylestrenol was shown by its effects on prostate volume (anatomy), maximum urinary flow rate (function), and symptom scores (symptom) at the end of 16 weeks of treatment. The rates of improvement for symptoms, QOL, function, and anatomy are 68.7% (N = 48), 79.2% (N = 48), 50.0% (N = 48), and 61.0% (N = 41), respectively. Overall efficacy (Good and Fair) was 70.9% (N = 48). During this study, 5 patients (3.9%) complained of loss of libido and 2 patients dropped out. In conclusion, allylestrenol was demonstrated to be a quite effective and safe medical treatment for patients with symptomatic BPH based on the criteria for treatment efficacy in BPH. PMID- 12094707 TI - [Clinical evaluation of the effect of tamsulosin hydrochloride and cernitin pollen extract on urinary disturbance associated with benign prostatic hyperplasia in a multicentered study]. AB - We evaluated the clinical efficacy and safety of tamsulosin hydrochloride and cernitin pollen extract in 243 patients with urinary disturbance associated with benign prostatic hyperplasia. They were assigned randomly to 3 groups, oral tamsulosin hydrochloride, cernitin pollen extract and their combination were administered for 12 weeks. The international prostate symptom score, post-voided residual urine and uroflowmetrogram were obtained before and after treatment. The international prostate symptom score improved in each group and then the maximum flow rate and average flow rate also increased significantly in the tamsulosin hydrochloride-administered groups. In conclusion, the administration of only tamsulosin hydrochloride and the combination of tamsulosin hydrochloride and cernitin pollen extract seemed more effective then the administration of only cernitin pollen extract in the treatment of urinary disturbance associated with benign prostatic hyperplasia. PMID- 12094709 TI - [Testicular sperm extraction for non-obstructive azoospermia: results with conventional and microsurgical techniques]. AB - Testicular sperm extraction (TESE) was performed on patients with non-obstructive azoospermia using the conventional or microdissection technique. First, conventional TESE was attempted on all patients. If the sperm was retrieved successfully, the TESE procedure was terminated. Microdissection TESE was indicated only for patients for whom the conventional sperm retrieval was unsuccessful. Sperm was successfully retrieved with conventional TESE from four (24%) of 17 patients with non-obstructive azoospermia. The remaining 13 patients underwent microdissection TESE and sperm was successfully retrieved from four of them. Application of the microdissection technique resulted in an improvement in sperm retrieval rates from 24% with conventional TESE to 48% when combined with the microdissection technique. None of the patients showed any acute or chronic complications after TESE. Intracytoplasmic sperm injection (ICSI) was performed on five of the eight patients with successful sperm retrieval and the partner of four of these patients became pregnant and progressed to delivery. These results indicate the usefulness of microdissection for improving the sperm retrieval rate in men with non-obstructive azoospermia. PMID- 12094710 TI - [A case of self-mutilation of testis]. AB - A 22-year old man was admitted to the emergency room of our hospital with bleeding from scrotal wound. He had resected his own scrotum and both testes by himself with a small knife, since he thought that his testes had developed necrosis. He had thrown his scrotal skin and both testes away in a lavatory. We closed the wide wound surface and the surrounding skin after ligation of the spermatic cord. The post-operative course was uneventful. The patient was referred to the psychiatric department of our hospital and received supportive psychotherapy under diagnosis of schizophrenia. This is the 23rd case to be reported as self-injury of the external genitalia and the 7th case limited to testis in the Japanese literature. PMID- 12094711 TI - [Saphenous vein grafting for Peyronie's disease: a case report]. AB - We present a case of Peyronie's disease in a 50-year-old male with a progressive, painful induration (3 x 2 x 0.5 cm) in the dorsal region of the penis. In order to correct the high degree of penile curvature, we made a venous patch graft from the saphenous vein under general anesthesia and straightened the penis. Six months after the operation, a recurrent induration was observed in the proximal region of the penis, and the penis became slightly shortened. PMID- 12094712 TI - [Schwannoma with purely cystic form originating from the adrenal area: a case report]. AB - A 62-year-old woman was hospitalized with complaints of right upper abdominal discomfort. Various imaging studies showed an extremely large suprarenal mass with solitary cystic formation. Partial adrenalectomy was successfully performed through the transperitoneal approach. The resected mass measured 12 x 10 x 10 cm and weighed 600 g. A pathological examination showed an Antoni-B predominant-type benign schwannoma containing a large volume of degenerative fluid. Our search of literature yielded few reports of solitary cystic schwannomas in the retroperitoneal cavity or throughout the body. This unusual cystic manifestation is thought to be a terminal stage of degeneration of a long-standing schwannoma. PMID- 12094713 TI - [A case of renal pelvic tumor due to phenacetin abuse]. AB - A 65-year-old female had been taking analgesics containing phenacetin, because of severe headaches since 1958. The total dose of phenacetin that she had taken was calculated to be 8.0 kg. She visited the department of urology in our hospital in August, 1999 complaining of gross hematuria. A solid mass was detected in her left renal pelvis on the abdominal computed tomographic (CT) scan. Under the diagnosis of a left renal pelvic tumor, nephrouretectomy was performed in September, 1999. Histopathological diagnosis was grade 2 transitional cell carcinoma. Interstitial nephritis was also observed. Our case is the twenty second report of an urinary tract tumor associated with phenacetin abuse in Japan. PMID- 12094714 TI - [Partial ureterectomy in a case of intrinsic-type ureteral endometriosis]. AB - We report a case of intrinsic-type ureteral endometriosis in a 28-year-old woman. She visited our hospital, complaining of lower abdominal pain. Drip infusion pyelography (DIP) showed left hydronephrosis, and left retrograde pyelography revealed a filling defect of the left pelvic ureter. Biopsy under ureteroscopy showed chronic ureteritis. It was difficult to exclude a malignant tumor, and we performed left partial ureterectomy and uretero-ureterostomy. The pathological examination showed endometrial tissue in the submucosal and muscle layer, and we diagnosed it as intrinsic-type ureteral endometriosis judging from the findings of the ureteroscopy and the operation. After the operation, drip infusion pyelography showed the improvement of the left hydronephrosis and ureteral passage, and the right ovarian endometriosis was found by gynecologists. She received hormonal therapy with the luteinizing hormone releasing hormone agonist for about half a year, but the right ovarian endometriosis remained unchanged. This is the ninth case in the Japanese literature reported as intrinsic-type ureteral endometriosis. PMID- 12094715 TI - Transitional cell carcinoma of the renal pelvis in a patient with cyclophosphamide therapy for malignant lymphoma: a case report and literature review. AB - Cyclophosphamide is considered to be a bladder carcinogen and there are many reports of secondary bladder cancer, while only a few cases of upper urothelial cancer have been described. A 59-year-old man, who had received cyclophosphamide therapy for malignant lymphoma, was suffering from gross hematuria and consulted our institute. Computerized tomography (CT), intravenous pyelography (IVP) and retrograde pyelography (RP) revealed a left renal pelvic tumor. Urinary cytology showed class V and radical left nephroureterectomy was performed. Histopathological diagnosis of the left renal pelvic tumor was transitional cell carcinoma, invading the renal parenchyma. He is free from recurrence eight months after surgery. To our knowledge, this is the third case of cyclophosphamide induced upper urothelial carcinoma reported in Japan, and the twelfth reported in the English literature. PMID- 12094716 TI - [Retroperitoneal malignant schwannoma: a case report]. AB - A 56-year-old woman was admitted with a complaint of a palpable left abdominal mass. The left kidney was compressed by the tumor which measured 18 x 10 cm in size on computed tomographic scanning. We diagnosed a left renal tumor or retroperitoneal tumor. No metastatic lesions were detected. We performed radical nephrectomy with complete tumor resection on August 28, 1997. The tumor had two satellites and was adherent to the renal capsule. A yellowish-white solid tumor was macroscopically encapsulated by fibrous tissue and had cystic lesions filled with bloody fluid, weighed 940 g and was 18 x 11 x 8 cm in size. Histopathological diagnosis was malignant schwannoma. No lymphnode metastases were observed. For poor responsiveness to chemotherapy and radiation therapy in this type of tumor, we did not perform adjuvant therapy. The patient is alive with no evidence of recurrence more than four years after surgery. PMID- 12094717 TI - [A case of extramammary Paget's disease presenting with anuria]. AB - A 64-year-old man was admitted with dyspnea and edema of the left lower extremity to the Department of Medicine, and referred to our department presenting with anuria and scrotal edema. Physical examinations revealed a large nodular tumor extending from his penis to the lower abdomen. He was diagnosed with penile tumor that had invaded into the retroperitoneal space. Histological findings revealed extramammary Paget's disease. Radiation therapy, low dose chemotherapy, and hormone therapy were performed, but he died of invasion and multiple metastasis of the tumor 5 months later. PMID- 12094718 TI - [A case of thyroid cancer metastasizing to the bilateral kidneys]. AB - A 66-year-old woman was admitted with a chief complaint of macroscopic hematuria. She had a past history of mediastinal tumor that had been surgically treated 11 years earlier and had been pathologically diagnosed as papillary thyroid cancer. Enhanced computed tomography demonstrated slightly enhanced renal tumors in both kidneys. Endoscopic findings showed bleeding from the left ureteral orifice. To control macroscopic hematuria, left nephrectomy was performed. Since, two months late severe bleeding occurred from the right ureteral orifice, right nephrectomy was performed and hemodialysis was initiated. The pathological findings of the bilateral renal tumors were papillary thyroid cancer suggesting metastases from the primary mediastinal tumor. PMID- 12094719 TI - [A case of retroperitoneal serous cyst with ossification]. AB - A 36-year-old woman came to our hospital complaining of right flank pain. Computed tomographic (CT) scanning showed a cystic mass, 6 x 9 cm in size, including homogeneous low-density fluid contents, in the right retroperitoneal space. The cyst wall showed partly high-density epithelium, but there was no contrast enhancement. A 7.5 x 12 cm retroperitoneal cyst was easily removed with yellow serous fluid in it. Cytological examination showed no malignant cells in this fluid. The origin of the cyst was unknown. The histopathological diagnosis was retroperitoneal serous cyst with focal ossification in the lining epithelium. Here we report this rare case of retroperitoneal serous cyst and briefly discuss 57 cases reported in Japan. PMID- 12094720 TI - [A case of cecovesical and rectovesical fistulas secondary to mucinous adenocarcinoma of the cecum]. AB - A hundred and nine cases of cecovesical fistula are described in the literature. Although its origin is usually reported to be appendicitis, we present a rare case of cecovesical and rectovesical fistulas secondary to mucinous adenocarcinoma of the cecum. An 84-year-old man was referred to our hospital with the chief complaints of fever and cloudy urine. An enterovesical fistula was seen on cystography, and cystoscopy showed a lot of mucinous fluid in the bladder and the papillary tumor arising from the enteric mucosa was identified through the fistula. Transurethral biopsy of the tumor revealed a mucinous adenocarcinoma of suspected colonic origin. At the operation, cecovesical and rectovesical fistulas secondary to cecal tumor were removed by right hemicolectomy, low anterior resection of the rectum and partial cystectomy. PMID- 12094721 TI - [Malignant fibrous histiocytoma of perirenal tissue: a case report]. AB - A 75-year-old woman was admitted to our hospital with an extremely large retroperitoneal tumor that had been detected with ultrasound on a routine health check. She had no complaint except lumbar pain. Computed tomography revealed a heterogenous tumor located outside the right kidney which was enhanced gradually. Doppler ultrasound showed mild vascularity in the tumor. We performed tumorectomy and right nephrectomy because the tumor was adherent to the right kidney. The tumor was 16 x 11 x 7 cm in size and weighed 621 g. The histopathological diagnosis was malignant fibrous histiocytoma. The tumor was considered to have arisen from perirenal tissue. PMID- 12094722 TI - Protobranch bivalves. AB - The subclass Protobranchia comprises more than 600 species of bivalves that occur throughout the world ocean. Mostly deposit feeders in soft sediments, they are abundant in the deep sea. Apomorphies that unite them as a group include gill structure, hinge conformation, shell microstructure, larval development, foot morphology, respiratory pigments, trophic mode and digestion. They are relatively small and highly conserved in form, originating in the Cambrian era. They may represent an ancestral, derived or paraphylectic group of the Bivalvia. The protobranchs include two orders, the Nuculoida and Solemyoida, which previously were classified separately in the subclasses Paleotaxodonta and Cryptodonta, respectively. They are of ecological interest and have a unique functional morphology. They feed mostly under the surface of the sediment with highly modified labial palps, but the degree to which they are selective in diet remains difficult to determine. They are important bioturbators in many soft-sediment assemblages; their feeding and locomotion affects sediment structure and community development. Solemyoids are unusual in inhabiting reducing environments and hydrocarbon seeps and in deriving their nutrition from endosymbiotic chemosynthetic bacteria. A variety of species of protobranchs are found in oceanic trenches, near hydrothermal vents, and in submarine caves. Protobranchs produce a lecithotrophic larval stage, the pericalymma, making their development unique among bivalves. The pericalymma remains in the plankton for a short time and presumably has low dispersal ability. Recruitment may be intermittent. Growth is rapid in post-larvae but decreases with age, though rates may not necessarily be slow, especially in continental shelf species. Life spans are commonly 1 to 2 decades, but deep-sea representatives may grow more slowly and live longer. Bottom fish, seastars and gastropods are their major predators and a few parasites and commensals have been documented. The predominance of protobranchs in deep-sea sediments may be a result of deep-sea origin or displacement from shallow waters by lamellibranchs. Their ability to deposit-feed, digest food extracellularly, and develop by means of lecithotrophic larvae make them particularly well adapted to cold and oligotrophic habitats. PMID- 12094723 TI - The Scaphopoda. AB - The Scaphopoda are marine infaunal carnivores that feed on foraminiferans and other microorganisms selected and manipulated by their unique feeding tentacles or captacula. Their tusk-like shell is open at both ends; the burrowing foot and captacula protrude anteriorly, while respiratory currents pass through the posterior opening. Although the scaphopods comprise one of the smallest molluskan classes in terms of species diversity, they have a worldwide distribution ranging from intertidal to depths in excess of 6000 m. Despite detailed monographic work from the late nineteenth and early twentieth centuries, however, the biology of scaphopods is today among the least understood of mollusks. To some extent, this is related to a significant departure from more typical molluscan organization inferred from studies of the larger classes. For example, the mantle cavity is elongate and restricted in space, and several associated organs are lost, including the osphradium and ctenidia; the latter is associated with the loss of auricles from the scaphopod heart. There is also little record of ontogeny in the adult shell, as the older portion is periodically decollated to maintain the posterior aperture for passage of inhalant and exhalant currents. This and other constraints on scaphopod diversification are reflected in the range limits of shell shape within the class. In contrast, the "Dentalium" egg has been a model system for experimental embryology, particularly in studies of cell lineage and early morphogenesis. Later larval development and most organogenesis is nonetheless poorly known, although recent studies have considerably enhanced our knowledge in this area. Global biogeographic patterns of scaphopod diversity have been studied only preliminarily. A general decrease in diversity with depth is broken by small diversity peaks at bathyal depths. There is also evidence for a marked latitudinal diversity gradient in the world's oceans, near equatorial in the Pacific but at approximately 20 degrees N in the Atlantic. Scaphopods have a wide diversity of ecto- and endo-symbiotic associations with other organisms, including commensal bacteria and ciliates, mutualistic anemones and corals, and parasitic algae and platyhelminths. Other documented associations include predation by naticid gastropods and ratfish, and the inhabitation of empty scaphopod shells by a variety of sipunculans and hermit crabs. Phylogeny within the class is still unresolved, although significant progress has been made recently in documenting morphological variation among families, genera, and species for application in a cladistic context. The evolutionary relationships of Scaphopoda within Mollusca are farther from resolution, with every conchiferan class a proposed sister group among modern analyses. Molecular avenues should provide significant progress in scaphopod phylogenetics, as indicated by recent comparative sequence analysis and gene expression patterns. Scaphopods comprise the most recent class of mollusks to appear in the fossil record, dating at least from the Mississippian Carboniferous (362.5 My). They appear from first-order occurrence data to have a substantially documented fossil record, with several sharp extinction and radiation events evident during their comparatively short geologic history. The unresolved position of the Scaphopoda within molluscan phylogeny, and their relatively limited range of morphological disparity but substantial derivation from other conchiferans, renders the scaphopods an enigmatic group. They represent a significant evolutionary avenue within the Mollusca, but one that is poorly examined. Nonetheless, comprehensive phylogenetic analysis and documentation of diversity within this small, monophyletic, class-level taxon, well-preserved in the fossil record, is imminently tractable, and presents a singular opportunity in the study of molluscan evolution. PMID- 12094724 TI - Pleurotomarioidean gastropods. AB - Pleurotomarioidean gastropods are continuously present in the fossil record since the Upper Cambrian and survive into the Recent fauna, thus providing rare insights into the evolutionary history of the class Gastropoda. Pleurotomarioidea achieved greatest numerical and morphological diversity during the Paleozoic, and dominated global shallow water marine gastropod faunas during the Paleozoic and Mesozoic. Only a single family, the Pleurotomariidae, survived the end-Cretaceous Extinction, but was restricted to deep water through most of the Cenozoic. The first living pleurotomariid was discovered during the mid-nineteenth century, along the bathyal zone of the western Atlantic. Subsequently collected specimens of these "living fossils" revealed that these animals comprise a mosaic of primitive and highly derived characters that originally inspired a new model of gastropod evolution, but more recently defy the orderly inclusion of this group within a cladistic framework of gastropod phylogeny. Molecular studies have likewise shown that the 18S rDNA gene of pleurotomariids contains novel insertions, and evolves more rapidly than in related taxa. A number of studies confirm that the Pleurotomarioidea may be included in the clade Vetigastropoda together with the Trochoidea, Fissurelloidea, Haliotoidea, and Scissurelloidea. However, neither the position of the superfamily within Vetigastropoda, nor the position of Vetigastropoda within Gastropoda is yet robustly resolved. The anatomical and molecular data are reviewed; the latter used to produce a well resolved phylogeny of the genera within the family, and to justify the naming of the long-used informal grouping "Perotrochus Group B" as the genus Bayerotrochus. A review of the geographic and bathymetric distributions of pleurotomariids reveals that the higher taxa segregate bathymetrically, while the species within each genus generally segregate geographically, so that most species exist in allopartry. The diet of pleurotomariids is reviewed, based on direct observations as well as analyses of gut contents, and is shown to consist primarily of sponges, but may include stalked crinoids, octocorals and, under aquarium conditions, fish and clam tissue. Despite their thin and relatively fragile shells, Pleurotomariidae survive an extraordinary number of attacks by predators, primarily crustaceans and fish. Their hypobranchial glands are capable of rapidly secreting a white fluid that probably serves as a chemical defense to repel predators. Circumstantial evidence suggests that this defensive mechanism may have originated during the Paleozoic. Despite the great antiquity of this lineage, pleurotomariids possess a substantial number of morphological, molecular and ecological novelties. PMID- 12094725 TI - Shelled opisthobranchs. AB - In his contributions to the monographic series "Manual of Conchology", Henry Pilsbry reviewed the subgroup Tectibranchiata, comprising those opisthobranch snails that (at least primitively) still possess a shell (Pilsbry, 1894-1896). Exemplified by the Cephalaspidea (bubble shells), others included in this group at Pilsbry's time and since were Anaspidea (sea hares) and the shelled members of Notaspidea (side-gilled slugs) and Sacoglossa (leaf slugs). Pilsbry (and others since his time) considered tectibranchs to be the "root stock" from which more advanced gastropods such as Nudibranchia and Pulmonata were derived. Tectibranch systematics is firmly based on conchology and most species were originally described from empty shells. However, soft-anatomical characters were acknowledged quite early on as equally important in tectibranchs, due to the reduction of their shells and their evolutionary proximity to unshelled gastropods. Today, Tectibranchiata is not recognized as a natural taxon although the word "tectibranch" (like "prosobranch" and "mesogastropod") continues in vernacular use. Shelled opisthobranchs have been redistributed among various taxa, including several new ones--the unresolved basal opisthobranchs (Architectibranchia) and the "lower Heterobranchia", an enigmatic and currently much-studied group of families considered basal to all of Euthyneura (Opisthobranchia and landsnails (Pulmonata)). Despite their polyphyletic status, shelled opisthobranchs remain important subjects in evolutionary studies of gastropods--as the most basal members of nearly every opisthobranch clade and as organisms with mosaic combinations of primitive and derived features within evolutionary "trends" (e.g., loss of the shell, detorsion, concentration of the nervous system, ecological specialization, etc.). Although they play a pivotal role, the shelled opisthobranchs have received minimal attention in more comprehensive gastropod studies, often relegated to token representatives at the derived end of prosobranchs or at the basal end of nudibranchs. The choice of this representative in a larger study is critical if its morphology and/or molecules are to adequately exemplify a larger group. This review explores the shelled opisthobranchs, including their history, current status and presumed synapomorphies, and emphasizes the importance of anatomical data to our current understanding of these "transitional" forms. A synthetic phylogenetic analysis, based on a combination of characters used in four published phylogenies involving tectibranchs, shows the current state of our knowledge and emphasizes areas for future study. The results indicate that Opisthobranchia, Cephalaspidea and Sacoglossa are monophyletic taxa, and that Acteon, the traditional basal opisthobranch, is convincingly a lower heterobranch. In most of the resulting cladograms, Anaspidea formed a monophyletic group with Cephalaspidea, as did pleurobranchoidean Notaspidea with Nudibranchia (the latter recently named as Nudipleura Wagele and Willan, 2000). PMID- 12094727 TI - Translocation of colicin from the receptor to the inner cell membrane: function of the peptidoglycan layer. AB - Sensitivity of spheroplasts (prepared in two ways) of a colicin-sensitive strain, of colicin-resistant and of colicin-tolerant mutants and of strains immune to colicins E1 and E2 was estimated and compared. Generally, the removal of the peptidoglycan layer brought about a slight nonspecific support for colicin translocation across the cell wall in sensitive, tolB tolerant and immune bacteria. tolB spheroplasts were colicin E1-sensitive, but E2-insensitive. Spheroplasts were always fragile and lysed spontaneously, especially those produced by lysozyme. Bacteria carrying tolA, tolQ and tolR mutations kept their colicin insensitivity as spheroplasts, just as the resistant ones. Bacteria rendered colicinogenic and hence colicin-immune turned to high colicin sensitivity in spheroplast form. The results indicate a change in plasma membrane associated with the spheroplast formation. PMID- 12094729 TI - Copper complexes with bioactive ligands. Part II--Antifungal activity. AB - Antifungal activity of new copper(II) complexes of 2-methylthionicotinate (2 MeSNic) of the composition Cu(2-MeSNic)2(MeNia)(2).4H2O (where MeNia is N methylnicotinamide), and Cu(2-MeSNic)2(Nia)(2).2H2O (where Nia is nicotinamide) and Cu(2-MeSNic)2L2 (where L is isonicotinamide, iNia, or ethyl nicotinate, EtNic) were tested on various strains of filamentous fungi by the macrodilution method. Most sensitive against copper(II) adducts with bioactive ligands were Rhizopus oryzae and Microsporum gypseum (IC50 1.5-2.3 mmol/L). The adducts with Nia, MeNia and EtNic at 5 mmol/L induced morphological changes in growing hyphae of Botrytis cinerea, mainly their intensive branching attached to release of cytoplasm with partial growth inhibition. Inhibition of sporulation (> 90%) of Alternaria alternata by Cu(2-MeSNic)2.H2O was observed as a change in the color of the colonies. The highest resistance was marked by B. cinerea and Fusarium moniliforme (average IC50 values 4.25 and 3.13 mmol/L, respectively). The presence of all bioactive ligands in copper(II) complexes caused an increase in the inhibition effect against model fungi (except significant inhibition activity of EtNic on R. oryzae). PMID- 12094728 TI - An evaluation of the outer membrane charge and softness of Thiobacillus ferrooxidans by the Ohshima's electrophoretic model of a "soft" particle. AB - The surface charge of bacterial cells plays an important role in their interfacial physiology and adhesion to substrata mediated by the electrostatic double-layer interaction. The surface charge or potential of biological cells is generally calculated from the experimentally measurable electrophoretic velocity of these cells migrating in an external electric field, applying the well-known Smoluchowski equation which is valid for "hard" particles with a sharp interface. However, bacterial cells possessing a structured outer membrane of a finite thickness (dependent on the ionic strength and pH of the surrounding liquid medium) are expected to obey Ohshima's electrophoretic mobility equation derived recently for "soft" particles. The electrophoretic mobility of Thiobacillus ferrooxidans was measured here by the fully automated technique of electrophoretic light scattering, based on the proportionality between the mobility and the Doppler shift in the frequency of light scattered by electrophoresing cells. Agreement was obtained between the experimentally determined electrophoretic mobility expressed as a function of low ionic strength (60-6000 mumol/L) at different pH values and the best-fit theoretical predictions of the "soft" particle electrophoresis theory, which is better than in the case of applying the Smoluchowski formula. The best-fit surface-charge and softness parameters predict a rather rigid and low-charge outer membrane of the bacterium examined, as compared to the parameters obtained for other bacteria in media of high ionic strength. PMID- 12094726 TI - ATP-dependent proteinases in bacteria. AB - Cytoplasmic proteolysis is an indispensable process for proper function of a cell. Degradation of many intracellular proteins is initiated by ATP-dependent proteinases, which are involved in the regulation of the level of proteins with short half-lives. In addition, they remove many damaged and abnormal proteins and thus play also an important role during stress. ATP-dependent proteinases are large multi-subunit assemblies composed of proteolytic core domains and ATPase containing regulatory domains on a single polypeptide chain or on distinct subunits, which can act as molecular chaperones. This review briefly summarizes the data about four main groups of these proteinases in bacteria (i.e. Lon, Clp family, HslUV and FtsH) and characterizes their structure, mechanism of action and properties. PMID- 12094730 TI - Regulation of the activity of intracellular alanylaminopeptidase synthesized by Pseudomonas sp. AB - Activity of purified alanylaminopeptidase of Pseudomonas sp. measured in the presence of the alanine derivative of 2-naphthoic acid (NA-Ala) is inhibited by 1,10-phenanthroline, EDTA, bestatin and amastatin; this finding supports the conclusion that this enzyme is a metallo-aminopeptidase. A decrease of its activity in the presence of iodoacetamide and its activation by thiols points to the significant role of -SH groups in the regulation of its activity. Co2+, Ca2+ and Mg2+ ions increased the enzyme activity while Zn2+, Cd2+ and Pb2+ markedly inhibited the enzyme even at low concentrations. A high thermal stability of alanylaminopeptidase depended on the presence of 1 mmol/L Co2+ and of 1 mmol/L L cysteine in the incubation mixture. PMID- 12094731 TI - Oxidative stress-induced expression of catalases in Comamonas terrigena. AB - When grown under oxidative stress, catalatic as well as peroxidatic activity is increased in the Gram-negative bacterium Comamonas terrigena N3H. Two distinct hydroperoxidases were demonstrated by a specific staining. Based on their molar masses and their sensitivity toward 3-amino-1,2,4-triazole and high temperatures, they were identified as dimeric catalase-1 (Cat-1; 150 kDa), and as a tetrameric catalase-2 (Cat-2; 240 kDa) with enhanced peroxidatic activity, respectively. These two catalases differ in their expression during the bacterial growth; whereas the expression of the smaller enzyme (Cat-1) is induced by 0.5 mmol/L peroxides in the medium, and to a lesser degree by 25 mg/L Cd2+, Cat-2 (typical catalase) is almost specifically induced with cadmium ions. PMID- 12094732 TI - Microorganisms in a high altitude glacier ice in Tibet. AB - Eighty-one strains of viable microorganisms were recovered from 23 samples collected from Ice Core 3 of Malan Glacier (China, 91 degrees 45.3' E, 35 degrees 48.4' N) drilled at high altitude (5620 m). All the strains were prokaryotes--75 of bacteria (including spore-forming ones) and 6 of actinomycetes. The characteristic genera differ from those of Arctic and Antarctic ice, in which many fungi and algae are widely distributed; this shows an difference of environmental conditions between Tibet and polar regions. The variation in number and species of Bacillus in different ice core layers implied changes of environmental conditions in the past. PMID- 12094734 TI - Mycoremediation of PAH-contaminated soil. AB - Out of a number of white-rot fungal cultures, strains of Irpex lacteus and Pleurotus ostreatus were selected for degradation of 7 three- and four-ring unsubstituted aromatic hydrocarbons (PAH) in two contaminated industrial soils. Respective data for removal of PAH in the two industrial soils by I. lacteus were: fluorene (41 and 67%), phenanthrene (20 and 56%), anthracene (29 and 49%), fluoranthene (29 and 57%), pyrene (24 and 42%), chrysene (16 and 32%) and benzo[a]anthracene (13 and 20%). In the same two industrial soils P. ostreatus degraded the PAH with respective removal figures of fluorene (26 and 35%), phenanthrene (0 and 20%), anthracene (19 and 53%), fluoranthene (29 and 31%), pyrene (22 and 42%), chrysene (0 and 42%) and benzo[a]anthracene (0 and 13%). The degradation of PAH was determined against concentration of PAH in non-treated contaminated soils after 14 weeks of incubation. The fungal degradation of PAH in soil was studied simultaneously with ecotoxicity evaluation of fungal treated and non-treated contaminated soils. Compared to non-treated contaminated soil, fungus treated soil samples indicated decrease in inhibition of bioluminescence in luminescent bacteria (Vibrio fischerii) and increase in germinated mustard (Brassica alba) seeds. PMID- 12094733 TI - Characterization of polychlorinated biphenyl-degrading bacteria isolated from contaminated sites in Czechia. AB - Biphenyl-utilizing polychlorinated biphenyls (PCB)-degrading bacteria were isolated from sites highly contaminated by PCBs, and their degradation abilities were determined using GC for typical commercial PCB mixtures (Delor 103 and Delor 106). Out of twelve strains which utilized biphenyl as a sole source of carbon and energy, strains Pseudomonas alcaligenes KP2 and P. fluorescens KP12, characterized by the BIOLOG identification system and the NEFERM test, were shown to significantly co-metabolize the PCB mixture Delor 103. DNA-DNA hybridization was used to compare both strains with well-known PCB-degraders Burkholderia cepacia strain LB400 and Ralstonia eutropha strain H850. The strain KP12 employs the same meta-fission route for degradation of chlorobenzoates as a chlorobiphenyl degrader Pseudomonas cepacia P166. Both isolates KP2 and KP12 belong to different phylogenetic groups, which indicates that the same geographical location does not ensure the same ancestor of degradative enzymes. We confirmed that also highly chlorinated and the most toxic congeners, which are contained in commercial PCB mixtures, can be biotransformed by members of indigenous bacterial-soil community under aerobic conditions. PMID- 12094736 TI - Yeasts and fungi in excrements of the sand hill snail, Theba pisana. AB - Eight microbial species were isolated and identified from excrements of the sand hill snail, Theba pisana, viz. 3 filamentous fungi, 3 basidiomycetous yeasts and 2 yeast-like ascomycetous fungi. PMID- 12094735 TI - GC clusters and the stability of mitochondrial genomes of Saccharomyces cerevisiae and related yeats. AB - The occurrence of GC clusters in Saccharomyces spp. and related yeasts was examined to clarify their association with the stability of intact mitochondrial genome. Abundance of nonspecific or specific GC clusters in these species decreases with phylogenetic distance from S. cerevisiae. Their number but not the number of replication origins correlates with the ability to form respiration deficient mutants induced by ethidium bromide. This effect is not associated with the nuclear background since the cybrids having identical nuclei and mitochondria from different species gave similar results. In contrast to grand genomes, the presence of GC clusters in rho- mutants does not play any role in ethidium bromide induced mtDNA loss. The most plausible explanation for mitotically lost petite mtDNA seems to be dilution during the distribution. PMID- 12094737 TI - Decolorization of industrial dyes by a Brazilian strain of Pleurotus pulmonarius producing laccase as the sole phenol-oxidizing enzyme. AB - The ability of a Brazilian strain of Pleurotus pulmonarius to decolorize structurally different synthetic dyes (including azo, triphenylmethane, heterocyclic and polymeric dyes) was investigated in solid and submerged cultures. Both were able to decolorize completely or partially 8 of 10 dyes (Amido Black, Congo Red, Trypan Blue, Methyl Green, Remazol Brilliant Blue R, Methyl Violet, Ethyl Violet, Brilliant Cresyl Blue). No decolorization of Methylene Blue and Poly R 478 was observed. Of the four phenol-oxidizing enzymes tested in culture filtrates (lignin peroxidase, manganese peroxidase, aryl alcohol oxidase, laccase), P. pulmonarius produced only laccase. Both laccase activity and dye decolorization were related to glucose and ammonium starvation or to induction by ferulic acid. The decolorization in vivo was tested using three dyes--Remazol Brilliant Blue R, Trypan Blue and Methyl Green. All of them were completely decolorized by crude extracellular extracts. Decolorization and laccase activity were equally affected by pH and temperature. Laccase can thus be considered to be the major enzyme involved in the ability of P. pulmonarius to decolorize industrial dyes. PMID- 12094738 TI - Production of lytic enzymes by Trichoderma spp. and their effect on the growth of phytopathogenic fungi. AB - The production of beta-1,3-glucanases and chitinases by three strains of Trichoderma in submerged cultures was determined. The synthesis of enzymes was induced by cell wall biopolymers of phytopathogenic fungi (Botrytis cinerea, Fusarium culmorum and F. oxysporum). T. hamatum produced the highest beta-1,3 glucanase activity; the most effective inducer of enzyme synthesis was the biomass of F. oxysporum. All examined strains of Trichoderma inhibited phytopathogen growth in biotic tests. The diffusion tests showed that the lytic enzymes take part in growth inhibition of phytopathogenic fungi. PMID- 12094739 TI - Decolorization of synthetic textile dyes by lignin peroxidase of Phanerochaete chrysosporium. AB - Neem hull waste (containing a high amount of lignin and other phenolic compounds) was used for lignin peroxidase production by Phanerochaete chrysosporum under solid-state fermentation conditions. Maximum decolorization achieved by partially purified lignin peroxidase was 80% for Porocion Brilliant Blue HGR, 83 for Ranocid Fast Blue, 70 for Acid Red 119 and 61 for Navidol Fast Black MSRL. The effects of different concentrations of veratryl alcohol, hydrogen peroxide, enzyme and dye on the efficiency of decolorization have been investigated. Maximum decolorization efficiency was observed at 0.2 and 0.4 mmol/L hydrogen peroxide, 2.5 mmol/L veratryl alcohol and pH 5.0 after a 1-h reaction, using 50 ppm of dyes and 9.96 mkat/L of enzyme. PMID- 12094740 TI - Efficacy of alkaloid (-)-corypalmine against spore germination of some fungi. AB - Inhibition activity of the alkaloid (-)-corypalmine on spore germination of plant pathogenic and saprophytic fungi (Alternaria solani, A brassicicola, A. brassicae, A. melongenae, Curvularia pallescens, C. lunata, C. maculans, Curvularisa sp., Colletotrichum sp., Helminthosporium speciferum, H. frumentacei, H. pennisetti, Heterosporium sp., Penicillum sp., Ustilago cynodontis) was determined. Spore germination of all the tested fungi was inhibited, Heterosporium sp. and Ustilago cynodontis being the most sensitive (complete inhibition of spore germination was observed at the very low concentration of 200 ppm). Curvularia palliscens, C. maculans and Curvularia sp. were less sensitive; complete inhibition of spore germination occurred at 400 ppm. PMID- 12094741 TI - Genetic heterogeneity of the pathogenic potentials of human and bovine group B streptococci. AB - One-hundred seventy-two B-streptococcal strains of human and bovine origin were analyzed for the presence of 9 genes potentially involved in virulence. Some of genes (glnA, cyl, hylB, scaA and cfb) were revealed in all the strains. However, the presence of others (bca, bac, scpB, lmb) varied from strain to strain. Taken together, 3 and 5 different types of pathogenic potential were found among human and bovine group B streptococci (GBS) strains, respectively, and only one type (bca+ bac scpB+ glnA+ cyl+ hylB+ lmb+ scaA+ cfb+) was common for both kinds of strains. We propose that different virulence genes can be involved in the development of infectious processes in humans and animals. A reliable PCR protocol with 3 pairs of primers (for the genes bca, bac and scpB) in the same reaction mixture was developed for the fast identification of the pathogenic potential of GBS. In comparison with the classical immunological methods this procedure displayed higher specificity and sensitivity as well as a shorter time of analysis. It can be recommended for use in the clinical and veterinary practice for studying the epidemiological relationship between the isolates and the ready identification of the clone causing the infection. PMID- 12094743 TI - Fungal diseases of the respiratory tract. AB - The proportion of Candida and non-Candida species in the clinical material from patients with respiratory-tract diseases was determined. C. albicans was isolated in 102 cases. An additional 89 strains of yeasts, isolated in association with respiratory diseases, belonged to 10 non-albicans Candida spp. and Cryptococcus spp. The prevailing species, which occurred in 47 cases, was C. parapsilosis. C. tropicalis, C. glabrata, and C. guilliermondii were isolated in 12, 10, and 9 cases, respectively. Four strains of C. krusei and three strains of C. lusitaniae and one strain each of C. freyschussii, C. robusta, C. zeylanoides, and Cryptococcus neoformans were also isolated. PMID- 12094742 TI - Detection of anti-Candida antibodies in neonates from a Neonatal Intensive Care Unit. AB - Anti-Candida antibodies were determined in a group of preterm neonates from a neonatal intensive care unit with serious diseases including candidemia. Antibodies to C. albicans blastospores, i.e. antibodies to C. albicans surface mannan and to C. albicans germ tubes were detected. Higher titers of antibodies to blastospores (1:320) occurred in all patients examined while antibodies to C. albicans germ tubes (with the highest titer of 1:160) were present in 32 out of 66 neonates examined. The highest titers of both anti-C. albicans blastospore antibodies and anti-C. albicans germ tube antibodies were detected in neonates with candidemia and disorders of saccharide metabolism. PMID- 12094744 TI - [Currently available blood saving methods in a patient at cardiac surgery under extracorporeal circulation]. AB - Different blood saving methods are analyzed in 2000 cardiac surgical patients undergoing coronary and vascular bypass surgery in 1993 to 2000. The basic blood saving methods are as follows: intraoperative autoreinfusion (normovolemic thermodilution), reinfusion of the patient's blood, preoperative autologous plasma donation in combination with aprotinine, aminocapronic acid, etc. An analysis revealed a decrease in homologous blood components intraoperatively. Red blood cell transfusion decreased from 100% in 1993 to 44% in 2000, fresh frozen plasma and platelet transfusions did from 98 to 39% and from 96 to 1%, respectively. Intraoperative homologous blood transfusion could be avoided in 70% of those undergone coronary bypass surgery. PMID- 12094745 TI - [Electrical stimulation of the spinal cord in cardiovascular disease]. AB - The early and late outcomes of electrical stimulation of the spinal cord are analyzed in chronic critical ischemia of the lower extremities and coronary heart disease with peripheral involvement of coronary arteries. The clinical capacities of this treatment that has enjoyed little use in our country due to its high cost showed atraumaticity of electrical stimulation of the spinal cord; a few cases of complications and no hospital mortality; a rapid positive clinical effect after the initiation of electric neurostimulation, and better life quality in patients in the postoperative periods. PMID- 12094746 TI - [Hypothermal perfusion: protection or damage?]. AB - Ninety six patients were examined during operations on the open heart and great vessels: 25 perfusions were performed at a temperature of 30-32 degrees C; 41 perfusions at 26-29 degrees C; 10 at 23-26 degrees C; 20 at 12-14 degrees C. It was found that with superficial hypothermia and moderate PaO2, blood myoglobin (MG) release was minimal and the count and activity of platelets were optimal. The degree of myoglobinemia increased as PaO2 rose. As the body's temperature lowered, the blood concentrations of MG, its differences smoothed in the subgroups with different PaO2 values. Critical myoglobinemia (over 30 times higher than the baseline values) was noted in a group with superdeep cooling to a temperature of 14 degrees C. By taking into account the fact that the myocardium contains large quantities of MG, loss of this heme-containing protein involves myocardial blood supply disorders and hence decreased myocardial contractility. A considerable platelet loss entails higher postoperative hemorrhagic diathesis and requires efforts in correcting coagulopathies. PMID- 12094747 TI - [Predictors of sudden death in patients with acute myocardial infarction by the results of Holter ECG monitoring]. PMID- 12094748 TI - [Complications after tracheal cicatricial stenosis surgery]. AB - A total of 323 patients (195 males and 128 females) were treated at the Russian Surgery Research Center, Russian Academy of Medical Sciences in 1963 to January 2001. They underwent 692 surgical or endoscopic interventions. Surgical treatment included circular tracheal resection and stepwise T-tube tracheal plastic repair. Three hundred and three patients were discharged, 20 patients died. Hospital mortality was 6.6%. Poor postoperative outcomes and intraoperative complications were noted after 93 (13.4%) interventions. The causes of death were bleeding in 5 patients, respiratory failure in 4, tracheal rupture at bougienage in 3, intraoperative vascular damage in 3, posthypoxic brain edema in 2, thrombotic embolism in 2, and total bronchospasm after tracheal endoscopic bougienage. Care to patients with cicatricial tracheal stenosis is frequently fraught with a risk and requires competence under the conditions of a specialized hospital. The most dangerous complications are those due to anastomotic incompetence, mediastinitis, bleeding, aspiration pneumonia, and respiratory failure. PMID- 12094749 TI - [Forecasting for and prospects of the development of surgery in 21st century]. PMID- 12094750 TI - [Reparative regeneration of the human liver grafted from a cadaver and a relative donor]. AB - The morphological features and dynamics of regeneration of the grafted human liver were studied by using data on 609 needle biopsies obtained for morphological monitoring in 38 recipients during transplantation of the liver from cadavers and relative donors. The biopsy specimens of donor organs taken prior to grafting served as a control. Irrespective of the type of grafting, regenerative processes in the grafted liver were shown to have common regularities and to run by using the mechanisms of both intracellular regeneration and proliferation. Their maximum rate was seen within a month after grafting of a cadaverous organ and within 3 months after grafting a part of the liver from a relative donor. In cadaverous transplantation, regenerative processes depended mainly on the degree of ischemic lesions; in relative donor organ transplantation, that depended on the fitness of graft mass (the index K being close to 0.5), proliferative processes and the time of higher hepatocytic ploidity increased. An adequate evaluation of the status of a cadaverous organ, estimation of the optimum mass of a grafted hepatic part from a relative donor, and effective immunosuppressive therapy ensure required reparation rates and a complete recovery of the structure of a graft. PMID- 12094751 TI - [Transplantation of the right hepatic lobe from an alive relative donor]. AB - The use of relative donors in the transplantation of the liver has shown a good performance as an alternative line in its orthotopical grafting. Shortage of donor organs actively stimulates the development of relative transplantation. The main problem of relative hepatic transplantation is the limited capacity of obtaining a required mass of a hepatic graft for obese recipients. To settle this problem, the Russian Surgery Research Center, Russian Academy of Medical Sciences, has developed an original safe procedure for obtaining the right lobe of the liver from an alive relative donor and for implanting it in a recipient. In November 1997 to October 2001, transplantation of the right hepatic lobe from an alive relative donor was made in 23 recipients (10 males and 13 females aged 9 to 55 (mean 22.3 +/- 3.1) years. Their body weight was 24 to 80 (mean 51.4 +/- 3.0) kg. Indications for surgery were as follows: hepatic cirrhosis (HC) at the end stage of the Wilson-Konovalov disease (n = 10), primary sclerotic cholangitis (n = 4), HC of viral etiology (n = 3), Bailer's disease (n = 2), primary biliary HC (n = 2), HC in the presence of alpha 1-antitrypsin deficiency (n = 1), and secondary biliary HC (n = 1). The donors of the right lobe of the liver were recipients' mothers in 9 cases, their fathers in 6 cases, sisters in 2 cases, sons in 1 cases, their daughter, brother, aunt, cousin in 1 case each. The donors' age ranged from 19 to 49 (mean 37.9 +/- 1.4) years. The donors underwent right hemihepatectomy, complications were absent in them. There were early mortality among the recipients. Two patients died in the late postoperative period. The remaining 21 recipients were survivors and followed up for 1 to 48 (mean 14.9 +/- 2.9) months. Their life quality was good. The use of the right lobe of the liver from an alive relative donor is the optimum alternative to transplantation of the cadaverous liver and partially compensates the shortage of donor organs for children, adolescents, and adults. PMID- 12094752 TI - [Plasmapheresis for the treatment of reperfusion injury during renal transplantation: impact on early and late outcome]. PMID- 12094753 TI - [Limb microcirculation in diabetes. Macro- and micro-hemodynamic relationship]. PMID- 12094754 TI - [The role of the Russian Surgery Research Center, Russian Academy of Medical Sciences, in the postgraduate training of medical staff]. AB - The paper presents a comprehensive analysis of the experience gained by the Russian Surgery Research Center over 40 years in improving the qualification of physicians and middle-level medical workers. It outlines teaching methods, shows the efficiency of flexible and mobile technologies in improving the qualification of medical staff, which are based on an individual differential approach to each student and which are oriented to satisfy the needs of a specific health care facility that has sent a specialist to be trained, provides evidence for that it is expedient and effective to improve the qualification of medical staff on the basis of a research institution, by taking into account the fact that the students will introduce new methods and technologies into practical public health care. PMID- 12094755 TI - [Indications for carotid endarterectomy and its outcomes]. AB - The paper analyzes a correlation between the sonographic and morphological characteristics of the structure of atherosclerotic plaques and the severity of cerebrovascular insufficiency. This significantly expands indications for surgical treatment in patients with carotid atherosclerosis. In the authors' opinion, not only the degree of carotid stenosis, but also the structure of a plaque is an important criterion for assessing indications for surgical treatment. The outcomes of carotid endarterectomy are comparatively evaluated in relation to the mode of carotid plastic repair. A role of intraoperative cerebral oximetry is shown. PMID- 12094756 TI - Proportional hazards models with random effects to examine centre effects in multicentre cancer clinical trials. AB - In randomized clinical trials comparing treatment effects on diseases such as cancer, a multicentre trial is usually conducted to accrue the required number of patients within a reasonable period of time. The fundamental point of conducting a multicentre trial is that all participating investigators must agree to follow the common study protocol. However, even with every attempt having been made to standardize the methods for diagnosing severity of disease and evaluating response to treatment, for example, they might be applied differently at different centres, and these may vary from comprehensive cancer centres to university hospitals to community hospitals. Therefore, in multicentre trials there is likely to be some degree of variation (heterogeneity) among centres in both the baseline risks and the treatment effects. While we estimate the overall treatment effect using a summary measure such as hazard ratio and usually interpret it as an average treatment effect over the centre, it is necessary to examine the homogeneity of the observed treatment effects across centres, that is, treatment-by-centre interaction. If the data are reasonably consistent with homogeneity of the observed treatment effects across centres, a single summary measure is adequate to describe the trial results and those results will contribute to the scientific generalization, the process of synthesizing knowledge from observations. On the other hand, if heterogeneity of treatment effects is found, we should carefully interpret the trial results and investigate the reason why the variation is seen. In the analyses of multicentre trials, a random effects approach is often used to model the centre effects. In this article, we focus on the proportional hazards models with random effects to examine centre variation in the treatment effects as well as the baseline risks, and review the parameter estimation procedures, frequentist approach-penalized maximum likelihood method--and Bayesian approach--Gibbs sampling method. We also briefly review the models for bivariate responses. We present a few real data examples from the biometrical literature to highlight the issues. PMID- 12094757 TI - Techniques for incorporating longitudinal measurements into analyses of survival data from clinical trials. AB - This article reviews existing approaches for joint analysis of longitudinal measurements, possibly measured with error or incompletely observed, and event time data, possibly censored. The models take the form of selection or pattern mixture models; estimation proceeds via the EM algorithm or Bayesian sampling techniques. The models are compared, their estimation and inferential procedures described, and advantages and disadvantages noted. Examples are discussed from several disease areas, including cancer and AIDS. PMID- 12094758 TI - Partitioning and peeling for constructing prognostic groups. AB - We investigate methods for identifying groups of patients with different prognosis. Our focus is on procedures that yield interpretable descriptions for groups of patients and corresponding regions of the predictor space. These strategies include tree-based methods and techniques which construct a single prognostic group through 'peeling' or refining of a larger group patients. The peeling methods are relatively new and are developed in detail. Simulations and examples for describing groups of patients with poor outcomes using data from a clinical trial for patients with multiple myeloma are presented. PMID- 12094759 TI - Added value of three-way methods for the analysis of mortality trends illustrated with worldwide female cancer mortality (1968-1985). AB - Trends in mortality rates are usually presented per tumour site or per country without an overall analysis of the complete data encompassing all three aspects (tumour sites, countries, trends). This paper presents a methodology for such an overall analysis using three-way methods applied to a data set on female mortality rates for 17 tumour sites of 43 countries for the years 1968-1985. Multivariate techniques like biplots and three-mode principal component analysis within an overall three-way analysis-of-variance framework were used. We confirmed the known patterns of comparatively high mortality for women due to cancer of the bladder, intestines, pancreas, rectum, breast, ovary, skin and leukaemia and the relatively low mortality rates for liver cancer in Western and Northern Europe, the USA, Australia and New Zealand. Also, the reverse pattern was observed for Middle and Southern Europe, Hong Kong, Singapore, and in Japan, and in some but not all Latin American countries. The relatively mortality due to cancer was high in the lungs, mouth, larynx and oesophagus in the British Isles, but was much less in other European countries. Mortality due to cancer of the thyroid, uterus, gall bladder and stomach was high in Middle European countries, as was the case in Japan, Chile and Costa Rica. Rates were low for Southern European countries, North America, Australia and New Zealand. Specific deviating patterns in the data were the more rapidly decreasing mortality rates for stomach cancer in Chile and Japan and the more rapidly increasing mortality rates for lung cancer in the USA, Scotland and Denmark. In conclusion, using three-way methods, it was feasible to analyse the cancer mortality data in their entirety. This enabled the simultaneous comparison of trends in relative mortality rates between all countries due to all tumour sites, as well as the identification of specific deviating trends for specific tumour sites in specific countries. PMID- 12094760 TI - [The signalling cascade of fertilization]. AB - The early event of fertilization-induced egg activation is a mobilization of intracellular Ca2+ that originates from the sperm entry point and spreads through the entire egg cytoplasm. Recently, this process has been established to require the sequential activation of Src family kinases, phospholipase C gamma, and inositoltrisphosphate receptor of endoplasmic reticulum. This review summarizes recent findings concerning the signalling pathway of fertilization from sperm-egg interaction to the Ca2+ release with emphasis on the role of tyrosine kinases in the egg activation. PMID- 12094761 TI - [Mode of epithelium-fibroblast interaction in mixed heterotypic cell cultures]. AB - The interaction between epithelium (dog kidney epithelium MDCJ/clone 20) and fibroblasts (diploid human fibroblasts M19 and AG-1523) was studied in mixed heterotypic cell cultures. The mode of cell interaction depends on the manner of their collision. At collision of the epithelium lamella and the lateral side of fibroblast, the lamella was seen to creep under the lateral side to force back the fibroblast. At the frontal collision of epithelium and fibroblast lamellae, the mode of interaction depends on the local situation. With the presence of a free substratum around, the fibroblast formed a new lamella and moved aside from the place of collision. In the case, when the neighboring cells prevented fibroblast from moving, it migrated under the epithelium. In this work, we have first demonstrated the formation of specialized intercellular adhesions between epithelium and fibroblasts. The cultures were studied by phase contrast, interference reflection or video tape recording, using an image processing system (Hamamatsu). For studying adhesion, immuno-fluorescent methods were performed. PMID- 12094762 TI - [The molecular determinants in the serpentine type receptors, responsible for its functional coupling with the heterotrimeric G-protein]. AB - In the review, data of the literature and own results on the functional coupling between the serpentine type receptors and the heterotrimeric G-proteins are analyzed and summarized. The role of cytoplasmic loops and C-tail domain of the receptors in interaction with G-protein alpha-subunits of different types is discussed. On the basis of theoretical analysis it is shown that the second cytoplasmic loop and the proximal to the membrane segments of the third cytoplasmic loop, containing the main G-protein-coupled molecular determinants, have the cationic nature and can form the helical structures. A molecular model of signal transduction from the receptor to G-protein, based on the electrostatic interactions between the cytoplasmic loops of receptors and receptor-binding regions of G-proteins, is developed. PMID- 12094763 TI - [Differences in muscle thermostability between diploid and tetraploid species of green toads (Bufo viridis complex) of Middle Asia]. AB - Significant differences were found between diploid (Bufo viridis) and tetraploid (B. danatensis) toad species in their skeletal muscle thermostability. The lower muscle thermostability in tetraploids may be associated with the adaptation of B. danatensis to colder conditions of arid mountain area in the Middle Asia. PMID- 12094764 TI - [Significance of nitric oxide function in development of cardiac hypertrophy under condition of experimental renal hypertension]. AB - The development of cardiac hypertrophy was studied under condition of experimental renal hypertension on the rat. The number of cardiac nitric oxide synthase (NOS)-positive neurones increased simultaneously with the increase in NOS-activity in these neurones. A connection was found between the development of cardiac hypertrophy and the activity of NOS in cardiomiocytes. The involvement of NO in the development cardiac hypertrophy as auto- and paracrine regulator is supposed. PMID- 12094765 TI - [The "air pouch" model in mice and a study of the wound fluid proteolytic activity]. AB - The experimental model of a cutaneous wound in mice has been offered, aimed to study the proteolytic activity of the wound fluid produced at early stages of wound healing, and to examine the continuous inflammatory state. The presence of metalloproteinases MMP-2 and MMP-9 in the wound fluid matrix was found to correlate with the existence of neutrophils and macrophages in the tissue. PMID- 12094766 TI - [Phosphorylation and dephosphorylation of rna polymerase III holoenzyme are modifications regulating the level of transcription in vitro]. AB - Two subforms of RNA polymerase III-IIIa and IIIb--were identified in human placenta nuclei. These subforms differed in molecular weight of one subunit, and in buoyant density in glycerol concentration gradient. Protein kinase activity, which phosphorylates at least four subunits of RNA polymerase IIIa and three subunits of RNA polymerase IIIb in vitro, was copurified with both the subforms. Protein kinase activity was inhibited by wortmannin, a specific PI3-kinase inhibitor. RNA polymerase III dephosphorylation by alkaline phosphatase in vitro decrease the transcription level on specific Alu-template. The associated protein kinase was not able to phosphorylate dephosphorylated RNA polymerase IIIa and to restore the transcription level up to the control one. PMID- 12094767 TI - [Expression of T-cadherin in the rat carotid artery wall after balloon injury and in different rat organs]. AB - T-cadherin is an unusual glycosilphosphatidylinositol (GPI)-anchored member of the cadherin family of cell adhesion proteins. In contrast to classical cadherins, tissue distribution of T-cadherin so far remained unknown. We examined tissue distribution of T-cadherin in rats using Western blotting and immunohistochemical method. Our results show that T-cadherin is expressed in all types of muscles (cardiac, striated, and smooth muscles), in brain neurons, and spinal cord, in the vessel endothelium, at the apical pole of intestinal villar epithelium, in the basal layer of skin, and eosophagal epithelium. Blood-derived and lymphoid cells as well as connective tissue were T-cadherin-negative. The highest level of T-cadherin expression was revealed in the cardiovascular system. Although T-cadherin was detected in smooth muscle cells, its role in the intimal thickening and restenosis is not known. We examined T-cadherin expression within 1-28 days after balloon injury of rat left carotid arteries. T-cadherin expression was valued immunohistochemically with semiquantitative method. In uninjured arteries, T-cadherin was expressed in endothelial (vWF-positive) cells, and smooth muscle (alpha-actin-positive) cells (SMCs). After denudation of arterial wall, T-cadherin was present both in the media and neointima. We revealed dynamics of T-cadherin expression in the media of injured artery: an essential increase being registered at the stage of cell migration and proliferation in the media and neointima (1-7 days), followed by its decrease to the baseline level (10-28 days). The high upregulation of T-cadherin expression in the media and neointima during migration and proliferation of vascular cells after vessel injury enables us to suggest the involvement of T-cadherin in vessel remodeling after balloon catheter injury. PMID- 12094769 TI - [Serotypes of the lower ciliate Dileptus anser]. AB - The author's data on the serotype composition in various clones of Dileptus anser are reviewed. This species is a novel subject of choice for studying the serotype problem in ciliates. The results obtained for serotypes of D. anser demonstrated several features shared also by some higher ciliates of the genera Paramecium and Tetrahymena, i.e. the well known classical models for serotype research. At the same time, some particular data obtained for D. anser do not fit in with the generally accepted picture of relevant events. Special attention is paid to a poor knowledge of the sexual process in this ciliate, which obviously hampers the true understanding of many events. Nevertheless, D. anser may be regarded as a suitable model for studying such important problems of general biology as epigenetic variability or inheritance. PMID- 12094768 TI - [Activity of toxins produced by Pseudomonas syringae pv. syringae in model and cell membranes]. AB - We studied effects of toxins produced by a bacterium Pseudomonas syringae pv. syringae on the conductance of bilayer lipid membranes (BLM). The used toxins were as follows: syringopeptin 22A (SP22A), syringomycin E (SPE), syringostatin A (SSA), syringotoxin B (STB), and methylated syringomycin E (CH3-SRE). All toxins demonstrated channel-forming activity. The threshold sequence for toxin activity was SP22A > SRE approximately equal to SSA > STB > CH3-SRE, and this sequence was independent of lipid membrane composition, and NaCl concentration (pH 6) in the membrane bathing solution (in the range of 0.1-1.0 M). This sequence correlated with relative bioactivities of toxins. In addition, SRE demonstrated a more potent antifungal activity than CH3-SRE. These findings suggest that ion channel formation may underlie the bioactivities of the above toxins. The properties of single ion channels formed by the toxins in BLMs were found to be similar, which points to the similarity in the channel structures. In negatively charged membranes, bathed with diluted electrolyte solutions (0.1 M NaCl), the channels were seen to open with positive transmembrane potentials (V) (from the side of toxin addition), and close with negative potentials. In uncharged membranes the opposite response to a voltage sign was observed. Increasing the NaCl concentration up to 1 M unified the voltage sensitivity of channels in charged and uncharged membranes: channels opened with negative V, and closed with positive V. With all systems, the voltage current curves of single channels were similarly superlinear in the applied voltage and asymmetric in its sign. It was found that the single channel conductance of STB and SSA was higher than that of other toxin channels. All the toxins formed at least two types of ion channels that were multiple by a factor of either 6 or 4 in their conductance. The results are discussed in terms of the structural features of toxin molecules. PMID- 12094770 TI - [Effect of symbiotic algae on the photoaccumulation capacity of cells of the ciliate Paramecium bursaria]. AB - The character of the effect produced by symbiotic algae on photodependent behavior of their host ciliates, Paramecium bursaria, was determined. Partially alga-freed paramecia showed a reliable increase in the rates of photoaccumulation. The photoaccumulation rate gradually decreased with a further decrease in the number of zoochlorellae. Once the chlorophyll content fell down to 20-25 mu/l, the ciliates lost their capacity for photoaccululation. A mathematical model of photoaccumulation has been constructed. PMID- 12094772 TI - The impact of genomic studies on neuropsychopharmacology. Proceedings of the 2nd European College of Neuropsychopharmacology (ECNP) Workshop. March 15-18, 2001, Nice, France. PMID- 12094771 TI - Acne variants. PMID- 12094773 TI - [Reaction to, "Expensive supplies and the aged patient"]. PMID- 12094774 TI - [Reaction to, "Implants and their future"]. PMID- 12094775 TI - Proceedings of the 19th European Workshop on Cognitive Neuropsychology. Bressanone, 2001. PMID- 12094776 TI - Antimicrobial periodontal therapy. PMID- 12094777 TI - ACOG practice bulletin. Diagnosis and management of preeclampsia and eclampsia. Number 33, January 2002. American College of Obstetricians and Gynecologists. AB - Hypertensive disease occurs in approximately 12-22% of pregnancies, and it is directly responsible for 17.6% of maternal deaths in the United States (1,2). However, there is confusion about the terminology and classification of these disorders. This bulletin will provide guidelines for the diagnosis and management of hypertensive disorders unique to pregnancy (ie, preeclampsia and eclampsia), as well as the various associated complications. Chronic hypertension has been discussed elsewhere (3). PMID- 12094778 TI - Potential outcomes of type Is explained. Conditional accreditation twice in one accreditation cycle triggers PDA rule. PMID- 12094779 TI - Probing and overcoming a mould menace. AB - Thorough investigation of an environmental problem within Brockville General Hospital, Ontario, Canada, was vital. This special report is by William Roth, Chris Rahm and Bruce G. Fraser--all of Jacques Whitford Environment Limited. William Roth is the first point of contact for Jacques Whitford's hospital clients and is a project manager within the Environmental Engineering group of JWEL's Ottawa office. Chris Rahm is a member of the Hazardous Materials group in JWEL's Ottawa office and was the site supervisor for the mould abatement at BGH. Bruce Fraser is the corporate technical director of indoor air quality for JWEL and regularly provides technical assistance for mould related projects carried out by JWEL. PMID- 12094780 TI - Should you fight it out or walk away? AB - Consider the repercussions carefully before you take on a legal battle with your employer. It may be better to simply resign and try something new. PMID- 12094782 TI - Uncertainty in climate models. PMID- 12094781 TI - Re: Immunocytochemical demonstration of caseins in mouse mammary glands. PMID- 12094783 TI - Doubting Bose-Einstein condensation in helium. PMID- 12094784 TI - Dietary restriction and life-span. PMID- 12094785 TI - Ice patch discovery: a joint effort. PMID- 12094786 TI - [Too little scalp hair]. PMID- 12094787 TI - Developing public health indicators in complex emergency response. AB - During the past decade, indicators for the assessment, monitoring, and evaluation of services provided by humanitarian organizations to populations affected by complex emergencies (CEs) were developed to improve the effectiveness and accountability of humanitarian response. The quality of data used to develop individual indicators and their relationship to positive health outcomes varies greatly. This article states the essential characteristics necessary for the development and implementation of effective indicators in CE response and proposes the establishment of an evidence-based grading system. The importance of trend analysis and the modification or addition of various indicators and their thresholds, according to phase and location of CEs, are stressed. Limitations in the development, implementation, and interpretation of these indicators, including those outside of the organizations' control are discussed. More evidence-based research is needed as to the type and thresholds of indicators that lead to improved health outcomes in populations affected by CEs. The use of indicators by non-governmental organizations, and how they affect their program's decision-making in different phases and settings within CEs need further study. Finally, the establishment of a regulating body with the authority to enforce the attainment of standards by use of these indicators is necessary to avoid inappropriate humanitarian assistance causing loss of life in the future. PMID- 12094788 TI - Does sextant prostate biopsy provide adequate sampling for early detection of prostate cancer? PMID- 12094789 TI - Bacchic medicine. Wine and alcohol therapies from Napoleon to the French paradox. Preface. PMID- 12094790 TI - The influence of biomass temperature on biostabilization-biodrying of municipal solid waste. AB - A laboratory study was carried out to obtain data on the influence of biomass temperature on biostabilization-biodrying of municipal solid waste (initial moisture content of 410 g kg wet weight (w.w.)(-1)). Three trials were carried out at three different biomass temperatures, obtained by airflow rate control (A = 70 degrees C, B = 60 degrees C and C = 45 degrees C). Biodegradation and biodrying were inversely correlated: fast biodrying produced low biological stability and vice versa. The product obtained from process A was characterized by the highest degradation coefficient (166 g kg TS0(-1); TS0(-1) = initial total solid content) and lowest water loss (409 g kg W0(-1); W0 = initial water content). Due to the high reduction of easily degradable volatile solid content and preservation of water, process A produced the highest biological stability (dynamic respiration index, DRI = 141 mg O2 kg VS(-1); VS = volatile solids) but the lowest energy content (EC = 10,351 kJ kg w.w.(-1)). Conversely, process C which showed the highest water elimination (667 g kg W0(-1)), and lowest degradation rate (18 g kg TS0(-1)) was optimal for refuse-derived fuel (RDF) production having the highest energy content (EC = 14,056 kJ kg w.w.(-1)). Nevertheless, the low biological stability reached, due to preservation of degradable volatile solids, at the end of the process (DRI = 1055 mg O2 kg VS( 1)), indicated that the RDF should be used immediately, without storage. Trial B showed substantial agreement between low moisture content (losses of 665 g kg W0( 1)), high energy content (EC = 13,558 kJ kg w.w.(-1)) and good biological stability (DRI = 166 mg O2 kg VS(-1)), so that, in this case, the product could be used immediately for RDF or stored with minimum pollutant impact (odors, leaches and biogas production). PMID- 12094791 TI - Fractionation and characterisation of dissolved organic matter from composting green wastes. AB - A new fractionation procedure using membrane ultrafiltration (UF), followed by chemical characterisation--measurement of total organic carbon (TOC), chemical oxygen demand (COD) and organic nitrogen and spectroscopic study--was applied to aqueous extracts of composting green wastes. Three membranes of molecular weight (MW) cut-offs of 1, 10 and 100 kDa were used. The study demonstrated the first step of the transfer of organic matter from the solids to the aqueous biofilm surrounding the solids. The microbiological consumption of the dissolved organic matter mainly used molecules smaller than 1 kDa, while the aromatisation of the organic matter, observed after 100 days composting, involved molecules larger than 10 kDa. PMID- 12094792 TI - Effect of carbon source on compost nitrogen and carbon losses. AB - The effect of C source on N losses by volatilization during composting was measured using four bulking agents, each at three humidity levels and composted in duplicate under passive and active aeration. The bulking agents were pine shavings alone and corrected with soybean, chopped grass hay alone and corrected with urea, long (unchopped) wheat straw and chopped oat straw. The readily available C of each bulking agent was determined by analyzing for BOD5. In 105 l laboratory vessels, the bulking agents were mixed with liquid swine manure and tap water for a C/N of 20 and three humidity levels of 60%, 65% and 70%. While being aerated actively or passively, the mixtures were composted for 21 days. Their initial and final C and N contents were measured to conduct a mass balance analysis and calculate C and N losses. C and N losses were compared to bulking agent BOD5. N losses were compared to C losses. The humidity level and aeration regime had no effect on C and N losses but the N losses were correlated to C losses and only the C losses could be correlated to the BOD5 of the bulking agent. Thus, the N losses are related not only to the availability of C but also to the extent of composting. A relationship established between N and C losses indicated that 85% of the initial total N of the compost was available for microbial degradation and that 70% of the available C was lost as CO2 during the immobilization process. PMID- 12094793 TI - A new procedure to produce lignocellulosic anion exchangers from agricultural waste materials. AB - Two lignocellulosic agricultural waste materials (LCM), sugarcane bagasse (BG) and rice hull (RH), were converted into weak-base anion exchanger and evaluated for their exchanger capacity for nitrate. Pure cellulose (PC) and pure alkaline lignin (PL) were also used as reference materials to elucidate possible reactivity in LCM. Epoxy and amino groups were introduced into BG, RH, PC and PL substrates after the reaction with epichlorohydrin and dimethylamine in the presence of pyridine and an organic solvent N,N-dimethylformamide (DMF). Amino group incorporation into cellulose decreased with the presence of water in the reaction mixture and increased with the reaction time and presence of a catalyst (pyridine). The highest maximum nitrate exchange capacity (Qmax) and yields of the prepared exchangers was obtained from PL (1.8 mmol g(-1) and 412.5%), followed by BG (1.41 mmol g(-1) and 300%), PC (1.34 mmol g(-1) and 166%) and RH (1.32 mmol g(-1) and 180%). The proposed synthetic procedure was effective in modifying PL, PC and LCM chemically resulting in a higher yield and nitrate removal capacity. PMID- 12094794 TI - Effect of UV radiation on thermotolerance, ethanol tolerance and osmotolerance of Saccharomyces cerevisiae VS1 and VS3 strains. AB - After a previous mass screening and enrichment programme for the isolation of thermotolerant yeasts, VS1, VS2, VS3 and VS4 strains isolated from soil samples, collected within the hot regions of Kothagudem Thermal Power Plant, AP, India, had a better thermotolerance, osmotolerance and ethanol tolerance than the other isolates. Among these isolates VS1 and VS3 were best performers. Efforts were made to further improve their osmotolerance, thermotolerance and ethanol tolerance by treating them with UV radiation. Mutants of VS1 and VS3 produced more biomass and ethanol than the parent strains at high temperature and glucose concentrations. The amount of biomass produced by VS1 and VS3 mutants was 0.25 and 0.20 g l(-1) more than the parent strains at 42 degrees C using 2% glucose. At high glucose concentrations VS1 and VS3 mutants produced biomass which was 0.70 and 0.30 g l(-1) at 30 degrees C and 0.10 and 0.20 g l(-1) at 40 degrees C more than the parent strains. The amount of ethanol produced by the mutants (VS1 and VS3) was 8.20 and 1.20 g l(-1) more than the parent strains at 42 degrees C using 150 g l(-1) glucose. More ethanol was produced by mutants (VS1 and VS3) than the parents at high glucose concentrations of 5.0 and 6.0 g l(-1) at 30 degrees C and 13.0 and 3.0 g l(-1) at 42 degrees C, respectively. These results indicated that UV mutagenesis can be used for improving thermotolerance, ethanol tolerance and osmotolerance in VS1 and VS3 yeast strains. PMID- 12094795 TI - Stratification of solids, nitrogen and phosphorus in swine manure in deep pits under slatted floors. AB - Manure slurries stored in pits under slatted floors of both finishing and nursery barns were sampled at four different depths to study stratifications of total solids (TS) and nutrients (nitrogen and phosphorus), and to determine the relationship between the stratification of TS and nutrients. The results obtained can be used to improve the management and handling of swine manure in the under slat storage pits. A management scheme that can be adopted for both the finishing and the nursery barns' pits is the layer-by-layer harvesting of the manure. The thinner manure, which is lower in nutrients, can be spread on land near the production units in larger volumes or it may pumped to land remote from the production units without causing many clogging problems. The thicker manure, higher in nutrients, can be transported to land further away and spread in smaller volumes. The TS content of each stratum can be used to accurately estimate the nitrogen and phosphorus levels in the respective strata so that application rates can easily be adjusted accordingly during the time of land application. PMID- 12094796 TI - Nitrogen mineralisation potential in calcareous soils amended with sewage sludge. AB - Mineralisation of organic N is an important consideration when determining the annual amount of sewage sludge to be applied to agricultural soils. The mineralisation of sludge organic N was studied in two different textured soils (clayey and sandy soil) treated with aerobic and anaerobic sludge at two different rates (30 and 50 g sludge kg(-1) soil). The mineralisation of sludge organic N was determined during 20 weeks incubation period by analysis of inorganic N produced by a non-leached procedure. Sludge organic N mineralisation was influenced by soil type, organic N mineralisation being greater in the sandy soil (from 30% to 41%) than in the clayey soil (from 13% to 24%). Mineralisation rates decreased rapidly the first two weeks, followed by a slower decrease with time. Although total mineralisation increased with sludge addition rate, net mineralisation decreased with sludge addition rate, probably due to denitrificaton losses. The aerobically treated sludge gave higher mineralisation rates than the anaerobically treated one. The values of N0 and k for treated soils varied depending on the type of sludge and soil. PMID- 12094797 TI - Microbial detoxification of waste rubber material by wood-rotting fungi. AB - The extensive use of rubber products, mainly tires, and the difficulties to recycle those products, has resulted in world wide environmental problems. Microbial devulcanisation is a promising way to increase the recycling of rubber materials. One obstacle is that several microorganisms tested for devulcanisation are sensitive to rubber additives. A way to overcome this might be to detoxify the rubber material with fungi prior to the devulcanisation. In this study, 15 species of white-rot and brown-rot fungi have been screened with regard to their capacity to degrade an aromatic model compound in the presence of ground waste tire rubber. The most effective fungus, Resinicium bicolor, was used for detoxification of rubber material. Increase in growth of the desulfurising bacterium Thiobacillus ferrooxidans in presence of the rubber treated with Resinicium bicolor compared to untreated rubber demonstrated that detoxification with fungi is possible. PMID- 12094798 TI - Nitrogen transformations during biological aerobic treatment of pig slurry: effect of intermittent aeration on nitrous oxide emissions. AB - A laboratory scale aeration treatment system was built to study the fate of nitrogen during aeration of pig slurry. For each run evaluated, the nitrogen mass balance was determined including measurement of the nitrous oxide gas emissions. Intermittent aeration led to a nitrogen removal of about 53% of the total nitrogen content of the raw slurry. About 18% of the total nitrogen content of the raw slurry was emitted as N2O during aeration with an aerobic to anoxic ratio equal to 0.625. In contrast, the extension of the anoxic period (aerobic to anoxic ratio = 0.375) allowed complete denitrification and avoided N2O emissions. PMID- 12094799 TI - Microbial production of extra-cellular phytase using polystyrene as inert solid support. AB - Aspergillus ficuum TUB F-1165 and Rhizopus oligosporus TUB F-1166 produced extra cellular phytase during solid-state fermentation (SSF) using polystyrene as inert support. Maximal enzyme production (10.07 U/g dry substrate (U/gds) for A. ficuum and 4.52 U/gds for R. oligosporus) was observed when SSF was carried out with substrate pH 6.0 and moisture 58.3%, incubation temperature 30 degrees C, inoculum size of 1.3 x 10(7) spores/5 g substrate, for 72 h for A. ficuum and with substrate pH 7.0 and moisture 58.3%, incubation temperature 30 degrees C, inoculum size of 1 x 10(6) spores/5 g substrate for 96 h for R. oligosporus. Results indicated scope for production of phytase using polystyrene as inert support. PMID- 12094800 TI - High-rate composting-vermicomposting of water hyacinth (Eichhornia crassipes, Mart. Solms). AB - In an attempt to develop a system with which the aquatic weed water hyacinth (Eichhornia crassipes, Mart. Solms) can be economically processed to generate vermicompost in large quantities, the weed was first composted by a 'high-rate' method and then subjected to vermicomposting in reactors operating at much larger densities of earthworm than recommended hitherto: 50, 62.5, 75, 87.5, 100, 112.5, 125, 137.5, and 150 adults of Eudrilus eugeniae Kinberg per litre of digester volume. The composting step was accomplished in 20 days and the composted weed was found to be vermicomposted three times as rapidly as uncomposted water hyacinth [Bioresource Technology 76 (2001) 177]. The studies substantiated the feasibility of high-rate composting-vermicomposting systems, as all reactors yielded consistent vermicast output during seven months of operation. There was no earthworm mortality during the first four months in spite of the high animal densities in the reactors. In the subsequent three months a total of 79 worms died out of 1650, representing less than 1.6% mortality per month. The results also indicated that an increase in the surface-to-volume ratio of the reactors might further improve their efficiency. PMID- 12094801 TI - Long term effect of municipal waste disposal on soil properties and productivity of sites used for urban agriculture in Abakaliki, Nigeria. AB - An assessment on the impact of long term (20 years) municipal waste disposal on soil physicochemical properties and soil productivity in municipal waste dump sites was carried out in urban Abakaliki, southeastern Nigeria. Profile pits were dug at selected points and auger and core samples were collected from different soil horizons of dump and non-dump sites using free survey technique to choose observation points. There were differences in particle size distribution between dump and non-dump sites. Similarly, soil bulk density was lower by 9-13% while total porosity and hydraulic conductivity were higher by 9-14% and 240-463%, respectively, in the dump site relative to non-dump sites. Soil organic matter, total nitrogen, cation exchange capacity (CEC) and percent base saturation increased from 701 to 743, 646 to 740, 7% to 10% and 5% to 14%, respectively, in the dump site soil relative to non-dump site soil. These results were confirmed by a high coefficient of alienation (degree of unrelatedness, square root(1 - R2)) in bulk density, 0.65; total porosity, 0.58; hydraulic conductivity, 0.87; organic matter, 0.93; total N, 0.82; pH, 0.85 and cation exchange capacity, 0.97 between dump and non-dump soils. Heavy metals (Pb, Cu, Fe and Zn) increased by between 214% and 2040% in dump site soils relative to non-dump site soil. This may lead to increased uptake of metals by some test crops although their transfer ratios differ from crop to crop. Long term dumping of municipal wastes can influence soil properties and productivity at municipal waste dump sites but still may be used for farming provided that ecotoxological risks associated with its usage are continuously assessed and controlled. PMID- 12094802 TI - Use of waste Chinese cabbage as a substrate for yeast biomass production. AB - The possibility of using waste Chinese cabbage (Brassica campestris) as a substrate for microbial biomass production was investigated. The juice from waste Chinese cabbage contains relatively high amounts of reducing sugars suitable for yeast culture. The cell mass and protein content of four species of yeast, Candida utilis, Pichia stipitis, Kluyveromyces marxianus, and Saccharomyces cerevisiae, were determined when cultured in juice extracted from cabbage waste. Compared to YM broth containing the same level of sugar, all the strains except C. utilis showed higher total protein production in cabbage juice medium (CJM). PMID- 12094803 TI - Effect of potassium salts and distillery effluent on carbon mineralization in soil. AB - Distillery effluent, a rich source of potassium, is used for irrigation at many places in the world. A laboratory experiment was conducted to study the influence of potassium salts present in post-methanation distillery effluent (PME) along with two other salts, KCl and K2SO4, on mineralization of carbon in soil. PME oxidized with H2O2, raw PME, KCl and K2SO4 solutions containing K equivalent to 10%, 20%, 40% and 100% of K present in PME were added to the soil separately, maintaining four replications for each treatment and control. Addition of salts up to a certain concentration stimulated C mineralization but a decline was noticed at higher concentrations. All the levels of salts caused higher CO2 evolution than the control suggesting that the presence of K salts enhanced the microbial activity resulting in increased CO2 evolution. The influence of K2SO4 was significantly higher than KCl in stimulating C mineralization in soil. Oxidized effluent had a higher stimulating effect than inorganic salts, showing the influence of other salts accompanying K in the PME. Raw PME, which contained excess organic C, increased CO2 evolution even at the highest salt level (100% PME) signifying the effect of added C on alleviating the salt stress on microbial activity. PMID- 12094804 TI - Effect of thermal and chemical pretreatments on anaerobic ammonium removal in treating septage using the UASB system. AB - This work investigated how thermal and chemical pretreatments influenced the anaerobic ammonium oxidation process in anaerobic digestion using biogranules that had been acclimated to septage. The digestion experiments were performed in serum vials using thermally and chemically pretreated septage as the substrate. The experimental results indicated that heat pretreatment reduced both dinitrogen production and ammonium removal. HCl and NaOH pretreatments increased both values by near 45% and 55% over the control. Alkali addition was more efficient than acid addition in enhancing dinitrogen production and ammonium removal. PMID- 12094805 TI - Comment on "Influence on different natural zeolite concentration on the anaerobic digestion of piggery waste" (Bioresource Technology 80 (2001) 37-43). PMID- 12094806 TI - [Remembrances of the professor Witold Rudowski (1918-2001)]. PMID- 12094807 TI - [Postgraduate training for doctors between the wars in Poland, 1918-1939]. AB - In the years between the Two World Wars in Poland, there have been made some meaningful changes in the approach towards the postgraduate education of the physicians. The widespread conviction about the need of widening the practical knowledge, was replaced in 1930 by a law, which made a year-long hospital practice obligatory for doctors who have accomplished their medical studies. The completion of this obligatory training was the condition of starting an independent professional practice. The aim of this complementary education, which comprised the basic branches of medicine, and which were conducted under the eyes of experienced doctors, was the complement of the theoretical academic knowledge by practical skills. The introduction of a law, which made a one-year-long postgraduate hospital practice obligatory, is undoubtedly one of the important achievements of the period between the Two World Wars, in the realisation of the idea of modern and complete education of the physicians in Poland. PMID- 12094808 TI - [Brain death: biological and ethical aspects]. AB - The article presents briefly historical development of death criteria from the modern times to the present. The criteria which are used for identification and diagnosing death on the base of respiratory and circulatory death definition are described. This work underlines the inadequacy of the definition of the brain death in relation to patients with persistent vegetative state and in relation to anencephalic newborns. The author describes the pathology and clinical and laboratory evidence of the brain stem death, which gave the possibility to justify the thesis that in case of the brain stem death ontological arguments are sufficient for diagnosing the death of a human being. The attention of the ethic of the life sanctity (on the base of halachic's law) and its opposing influence on the evolution of the medical definition of death has been paid. The recognition of the brain as the death of an individual is a cultural shock, which from scientific point of view changed the ways of thinking, almost immediately but did not in the awareness of the society. The work also underlies the fact that utilitarian argumentation can not be a criterion for making a decision concerning the life of an individual. PMID- 12094809 TI - [The surname of Adamkiewicz in the medical terminology in the last three decades of the 20th century]. AB - Nearly eighty years after his death, Albert Adamkiewicz (1850-1921) has still been persisting in both the history of medicine owing to his work and in the medical terminology owing to eponymy: since his flourishing period toward the end of the XIXth century, the surname Adamkiewicz has entered the language of science as a proper-name constituent of anatomical, pathological, neurological, surgical as well as orthopaedic terms, combing with the appellatives stain, corpuscle or demilune, reaction or test, serum, syndrome as well as artery. Estimation of the actual vitality of particular eponymous terms compared with non-eponymous synonyms had to be the aim of the presented search in the scientific literature a century after. In contrast with the inert non-periodical (encyclopaedic) literature, periodicals have revealed all the eponymous terms fallen in oblivion except the 'Adamkiewicz artery' that has only recently been introduced in encyclopaedias although constantly preferred in periodicals of the period under investigation (appearing in 75% articles) over the most frequent non-eponymous synonym 'arteria radicularis magna / great(er) radicular artery' (scarcely 11% articles). Thanks to the 'artery' - joining furthermore several synonyms to appear nearly in 86% articles altogether - the surname Adamkiewicz persists in the living language of science; that is why its bearer ought to be remembered and mentioned even on the threshold of the XXIst century. PMID- 12094810 TI - The role of private hospitals in the California healthcare safety net: a comparison of charitable contributions reported by for-profit and non-profit hospitals in 1998. PMID- 12094811 TI - Delays in treatment. PMID- 12094812 TI - Activation of the CheA kinase by asparagine in Bacillus subtilis chemotaxis. AB - Past experiments have shown that CheA and CheY are required to generate smooth swimming signals in Bacillus subtilis chemotaxis. This study, as anticipated from in vivo experiments, demonstrates in vitro that an attractant-bound chemoreceptor leads to an increase in CheA activity, which in turn leads to an increase in the CheY-P pool that ultimately causes a behavioural change in the bacteria. Asparagine has been found to increase the rate of CheY-P formation in the presence of McpB-containing membranes, CheA, and an excess of CheY. This asparagine effect requires the presence of both CheA and McpB, the latter of which has been shown to be the sole receptor for this attractant. Utilizing membranes from a number of B. subtilis null mutant strains, insight has also been gained into the potential roles of a number of unique chemotaxis proteins in the regulation of CheA activity in the presence and absence of this attractant. PMID- 12094813 TI - [A forgotten discovery: doctor of medicine Ernest Duchesne's thesis (1874-1912)]. AB - It was only in 1946 that Ramon and Richou's publication in the newspaper "le Progres Medical", reminded of Ernest Duchesne's thesis which dealt for the first time with antagonism between germs and mould like penicillium. Thanks to Justin Godard's paper at the Academie de Medecine, Duchesne was known as "the precursor of the antibiotic action of penicillium". From that moment, some papers were written and the thesis was even published in a luxurious edition. The 1983 year of the "Ecole du Service de Sante des Armees" was called Ernest Duchesne who can be considered as "the theoretician precursor of antibiotic therapy" according to Francois Chast. PMID- 12094814 TI - [Louis Maurice Champagne and the "Ecole dentaire francaise"]. AB - The Royal Edict of Louis XIV had created a new category of practitioners, "The Teeth Experts", and at the dawn of the second half of the 19th century, France had no dental school nor specific legislation. The situation worsened when many American dentists settled in France. Thus the creating of the "Ecole dentaire de Paris" in 1880 constituted a significant event which was followed by the opening of the "Ecole Dentaire de France" in 1884 and of the "Ecole Dentaire Francaise" in 1896. Louis Maurice Champagne will be its director from 1926. At the St Ouen Jules Valles Market, we fortuitously discovered a framed document containing three photographs of Louis Maurice Champagne's surgery located, 3 Place de la Madeleine in Paris. Thanks to his son and through various documents, we could trace the exceptional professional carrier of this practitioner who eventually bought the "Ecole Dentaire Francaise" from Dr Louis-Jules Rousseau. Then, we discuss the legal frame work of the 1892 law created the "Chirurgien-dentiste" diploma and we describe the scientific and practical teaching for the "Ecole dentaire francaise". The operating layout of the clinical rooms is presented by opposing the ergonomic aspect of the demonstration surgery of the director and the two large operative dentistry rooms used for students. We conclude by emphasizing Louis Maurice Champagne's determination to overcome adversity which was peculiar to him as a young orphan immigrant without resources. To solve the problem of the "Chirurgien Dentiste" statute considered by its promoters as a stage of transition towards fuller reforms, we present various projects like from Dr Hulin's. This practitioner presided the Dental Section of the Medical Council Order before being voted in the first President of the "Chirurgien Dentiste" National Council Order in 1945. In 1942 he was the first to advocate a legislation based on the independence of the dental profession, still in force nowadays. PMID- 12094815 TI - [Galvanic and Faradic power in Anaesthesia]. AB - Electrical anaesthetic methods have been applied for the first time in Paris (May June 1847) by the physiologist and anatomist scientist Fortune Christophe Ducros. Ducros's scientific research works were not published so they were forgotten. In fact the electrical anaesthetic methods were dropped on account of the difficult conceiving of the electrical devices and the easiness of methods using either and chloroform. In 1858 J.B. Francis, a dentist of Philadelphia, pulled out 164 teeth in presence of official dentists by means of this method. Apolloni Pierre Preterre, a Parisian dental surgeon, introduced again the method in June 1858 and Victor Auguste Francois Morel-Lavallee, Jean-Baptiste Fonssagrives and Leon Athanase Gosselin tested it in some French hospitals without further delay. In December 1859, Jean-Baptiste Georges invented a device to use galvanism and induction particularly for cauterisation in the mouth or for local anaesthesia. Eventually the electrical method failed and was neglected. PMID- 12094816 TI - [The Leroy de la Faudignere, a family of dentists of the 18th century]. AB - Francois Leroy de la Faudignere was the founder of the family. Although he was not an expert in dentistry, he acquired a good reputation of dentist in the second half of the 18th century. He published some books between 1766 and 1780. He lived in Paris and set up in 1780 at the Pavillon Royal, Plave Royale no1 (Place des Vosges nowadays). He was mainly busy with selling an approved analgesic elixir of his own. Out of his 5 children, 2 have been experts in dentistry and one of his daughters, Marie Marianne Francoise, married Jacques Rene Duval, Maitre en Chirurgie. Francois died six weeks after his daughter's marriage but her brothers sued her about the elixir their father had bequeathed to her. She won her case and carried on selling the elixir with Duval. Her daughter married Jean Nicolas Marjolin, a famous surgeon. Their son Rene Nicolas became a surgeon too. PMID- 12094817 TI - [Armauer Hansen (1841-1912), portrait of a Nordic pioneer]. AB - Desending from a Danish family, Gerhard Henrik Armauer Hansen was born in Bergan, Norway, on 29 July, 1841. Graduated in medicine in 1866 from the University at Christiana (the former name of Oslo), he started his medical career as an assistant physician at the National Hospital of Oslo for one year, working afterwards as a medical officer for a fishing company outside the Arctic Circle. Taking not well his routine job, he soon returned to Bergen where he began his work on a disease known as leprosy at the age of 26 and as an assistant of D.C. Danielssen at the Lungegaarden Hospital. There he engaged in research and investigations on the nature of the disease. Studying the various types of leprosy (skin, nerve and visceral lesions), he was awarded, one year after beginning his work, the gold metal of the University for his first publication on the disease. While Danielssen leaned toward heredity as a dominant factor in leprosy, Hansen's conviction was that the disease must have an infectious causal agent. In about 1871, Hansen began to observe tiny little rods in tissue specimens and considered they could be the ethiologic agent of leprosy, the more he found these rods in all the infiltrated nodular lesions in his patients. Finally, he proposed on February 28, 1873, that the rods were bacilli, responsible of leprosy. Promoted to the rank of Chief of the Leprosy Service in 1875, he held his job for 37 years; Hansen's entire adult life was spent at work in leprosy. He edited the journal <>. Hansen was also an eminent zoologist engaged in studies involving mollusks and worms; Since 1874, he was president of the Bergen Museum of Natural History. Armauer Hansen died on February 12, 1912, and the funeral ceremonies took place in the Museum of Bergen where his ashes are still kept. PMID- 12094818 TI - The duality of the brain and the multiplicity of minds: can you have it both ways? AB - Multiple Personality Disorder (MPD), now known as Dissociative Identity Disorder (DID), has been of great interest to the public for over a century. Case histories of MPD can be found in the literature as far back as the eighteenth century; nevertheless, publications from the latter part of the nineteenth century best describe this disorder as we know it today. This paper traces the case history literature of DID (MPD) from the earliest period to the present. This is done in such a way as to illuminate the basic theoretical and epistemological issues that are necessary to understand the process of dissociation (both normal, and abnormal aspects) and the role of hypnosis and its relationship to organic and 'hysterical epilepsy.' The theories of Fanet, Prince and Sidis are the major authorities discussed. The paper concludes with a discussion of the danger inherent in fostering a deterministic or reductionistic theory of consciousness. PMID- 12094819 TI - Focus on amisulpride. AB - Amisulpride is a second-generation antipsychotic, a substituted benzamide. It appears to be an effective agent in treating schizophrenia for what are characterised as positive and negative symptoms. The recommended doses are between 400 mg/day and 800 mg/day. Amisulpride demonstrates a good global safety profile, particularly when compared with first-generation antipsychotics, such as haloperidol. There are interesting studies that point towards amisulpride's antidepressant effect in dysthymia speculative on possible roles in affective psychoses and chronic fatigue syndrome. PMID- 12094820 TI - Association of apolipoprotein E polymorphism with myocardial infarction in Greek patients with coronary artery disease. AB - Studies in several populations have indicated that genetic variation at the apolipoprotein E (apoE) structural locus influences the risk of coronary artery disease (CAD) and myocardial infarction (MI). This study aimed at investigating whether apoE polymorphism has an allelic and/or genotypic impact on the risk of MI in Greek patients with CAD. We compared apoE gene polymorphism in a group of patients with angiographically confirmed CAD but not MI [CAD/MI (-)-group, n = 143] and a group of age and sex-matched CAD patients who had experienced a non fatal Ml [CAD/MI (+)-group, n = 124]. The patients were also compared with a group of healthy younger individuals (n = 240) with no family history of CAD. The apoE genotype distribution differed significantly between the two groups of CAD patients (p = 0.02). The epsilon2 allele was 5.3-fold less frequent in the CAD/ MI (+)-group compared with the CAD/MI (-)-group (1.2% vs. 6.3%, p = 0.01). The frequency of the epsilon2 allele in healthy subjects was 8.1%, which is 6.8-fold higher than in CAD/MI (+)-patients (p = 0.001) and twice as high compared with all CAD patients (p = 0.02). No differences in epsilon4 allele frequencies were observed between CAD/MI (+)- and CAD/MI (-)-patients (10.9% vs. 9.8%), or between patients with CAD and healthy subjects (10.3% vs. 10.2%). In summary, the epsilon4 allele was not found to be associated with an increased risk for CAD or MI. In contrast, a negative association of the epsilon2 allele with Ml was observed among Greek patients with CAD. PMID- 12094821 TI - Combined treatment with fibrates and small doses of atorvastatin in patients with mixed hyperlipidemia. AB - Combined statin and fibrate therapy is often imperative for the improvement of the serum lipid profile in patients with mixed hyperlipidemia. However, the potential risk of myopathy has limited the widespread use of such therapy. Preferably this treatment should involve low optimally tolerable doses of hypolipidemic drugs. Thus, we undertook a study to determine the safety and efficacy of combination therapy with fibrates and small doses of atorvastatin. Twenty-two patients with mixed hyperlipidemia were started on a fibrate regimen (micronised fenofibrate 200mg/day or ciprofibrate 100 mg/day). Because after 12 weeks of therapy the fibrate failed to normalise the serum lipid profile, small doses of atorvastatin (5 mg/day) were added for a further 12 weeks. The administration of the fibrates resulted in a significant decrease in total and LDL-cholesterol levels, as well as in triglycerides, and an increase in HDL cholesterol levels. The addition of atorvastatin (5 mg/day) resulted in a further decrease in total and LDL-cholesterol levels. Consequently, the hypolipidemic therapy target was achieved in most of the patents. Combination therapy was well tolerated and no significant increases in serum liver and muscle enzymes were noticed. We conclude that the careful administration of small doses of atorvastatin in patients with mixed dyslipidemia receiving fibrates is associated with a significant amelioration of lipid abnormalities. PMID- 12094822 TI - Efficacy and safety of rivastigmine in patients with Alzheimer's disease who failed to benefit from treatment with donepezil. AB - BACKGROUND: Selective acetylcholinesterase (AChE) and dual acetyl- and butyrylcholinesterase inhibitors constitute the only approved agents for the symptomatic treatment of Alzheimer's disease (AD). Donepezil is a specific, reversible inhibitor of AChE, while rivastigmine is a slowly reversible (pseudoirreversible) dual cholinesterase (ChE) inhibitor, with brain-regional specificity for the cerebral cortex and hippocampus. According to the European Marketing Authorisations, the clinical benefit of ChE inhibitors should be reassessed on a regular basis and discontinuation should be considered when evidence of a therapeutic effect is no longer present. However, substantial differences in the pharmacological and pharmacokinetic profiles of the available ChE inhibitors suggest that it may be desirable to switch between ChE inhibitors if patients fail to show efficacy, deteriorate or are unable to tolerate their initially prescribed medication. DESIGN: This open-label, six-month study evaluated the efficacy and safety of rivastigmine in 382 AD patients who had previously failed to benefit from treatment with donepezil (80% due to lack of efficacy, 11% due to tolerability problems, 9% both reasons). RESULTS: At the end of the study, 56.2% of patients were responders to rivastigmine, as assessed using a global function scale (the Clinicians' Global Impression of Change). Cognitive performance (measured by the Mini-Mental State Examination) and the ability to perform activities of daily living (measured by the Instrumental Activities of Daily Living scale) were improved/stabilised in 48.9% and 57.0% of patients, respectively. Rivastigmine was generally well tolerated, the most common adverse events being nausea and vomiting, consistent with reports from previous clinical studies. The occurrence of side-effects or lack of efficacy with donepezil treatment was not a predictor of similar problems when treated with rivastigmine. CONCLUSION: Rivastigmine treatment appears to be beneficial in AD patients who have previously failed to benefit from, or were unable to tolerate treatment with, donepezil. PMID- 12094823 TI - The role of losartan in cost-effective hypertension control. AB - Despite recent guidelines emphasising the need for aggressive treatment in patients with elevated blood pressure, the control of hypertension in Europe and the USA is poor, imposing a considerable burden in terms of patient morbidity and mortality, and associated healthcare costs. A major factor contributing to the suboptimal control of hypertension is the failure of patients to adhere to their prescribed therapy. Drug side-effects are an important cause of non-compliance and prescribing a well-tolerated agent that promotes good compliance is therefore the key to the cost-effective management of hypertension. Several studies have demonstrated that patients are more likely to remain on therapy with the angiotensin II antagonist losartan than other antihypertensives. Although the acquisition costs of new antihypertensives such as losartan are greater than for older drugs, such costs represent only a small proportion of the total cost of prescribing antihypertensive therapy. When accessory costs are also considered, the total cost of care with newer antihypertensives is comparable with those for diuretics. The costs involved if therapy has to be switched due to unacceptable side-effects also need to be taken into account when assessing relative cost effectiveness. Furthermore, savings may accrue from the non-haemodynamic benefits of losartan, such as improved cognitive function and renal protection. Further studies will increase awareness of the true cost effectiveness of antihypertensive drugs. PMID- 12094824 TI - Patient and physician satisfaction with aceclofenac: results of the European Observational Cohort Study (experience with aceclofenac for inflammatory pain in daily practice). Aceclofenac is the treatment of choice for patients and physicians in the management of inflammatory pain. AB - A pan-European study involving 23407 patients with pain due to various inflammatory or degenerative rheumatic diseases was undertaken in Austria, Belgium, Germany and Greece, to evaluate overall pain relief and satisfaction with aceclofenac therapy. Aceclofenac was considered by patients to be a highly efficacious treatment with excellent and fast analgesic activity that was maintained throughout the study period. At the conclusion of the study, assessment of patient status, a parameter encompassing both efficacy against inflammatory pain and tolerability, by both patient and physician, was either much improved or improved in 84% of cases. These evaluations were similar irrespective of the country or whether the indication was acute (e.g. post-pain) or chronic pain (e.g. osteoarthritis). Patient satisfaction with, and compliance of, aceclofenac therapy was similarly impressive; 90% of patients were satisfied and over 90% of patients were treatment compliant. In combination with the recently published SAMM study results, the findings of the European Observational Cohort study validate aceclofenac, in everyday clinical practice, as an effective, well-tolerated and well-accepted therapy for both acute and chronic inflammatory and degenerative disease. The availability of a powerful anti inflammatory agent with a low incidence of side-effects is of considerable value to both the patient and physician in the management of inflammatory pain. This objective has been fulfilled with aceclofenac therapy. PMID- 12094825 TI - The effect of fluvastatin on plasma fibrinogen levels. AB - There are conflicting data with regard to the effect of statins, including fluvastatin, on plasma fibrinogen levels. We undertook the present study to examine the influence of fluvastatin (40 mg/day) on plasma fibrinogen levels in hypercholesterolemic non-smoker patients with normal triglyceride levels (< 2.25 mmol/l) (n = 65). Fluvastatin administration was followed by a significant decrease of plasma fibrinogen levels by 8% (from 3.6 g/l to 3.33 g/l median value, p < 0.02). No correlation was found between the change in lipids and that in fibrinogen levels. Furthermore, there was no correlation between plasma fibrinogen levels and the change in fibrinogen concentration after fluvastatin administration. These results are in contrast with previously published data. However, the majority of the previous studies included patients with ischemic heart disease, unlike our study population. Furthermore, in some of these studies, which Included relatively small numbers of patients, there was a trend towards a decrease in plasma fibrinogen concentration after fluvastatin administration. We conclude that fluvastatin can significantly decrease plasma fibrinogen levels. However, more studies using fluvastatin and other drugs of this class are necessary to clarify this issue. PMID- 12094826 TI - Understanding and managing behavioural symptoms in Alzheimer's disease and related dementias: focus on rivastigmine. AB - Behavioural and psychological symptoms of dementia (BPSD) are among the most distressing manifestations of dementia and result in considerable social and economic costs. Practical, non-pharmacological approaches such as environmental and behavioural changes may provide some benefit for patients in managing mild BPSD. In addition, various pharmacological approaches to treatment have been employed, such as neuroleptics and atypical antipsychotics, which differ in neurochemical target and clinical effectiveness. Growing evidence suggests that the neurobiological basis of BPSD in Alzheimer's disease (AD) and related dementias is a loss of cholinergic neurones and a resultant decline in acetylcholine (ACh) in brain regions which regulate behavioural and emotional responses, such as the limbic system. This cholinergic deficit can be partly corrected by inhibiting cholinesterase enzymes (ChEs). Studies of ChE inhibitors have shown positive effects to improve or stabilise existing BPSD and delay the emergence of new behavioural symptoms. In placebo-controlled studies, donepezil has reported efficacy in non-institutionalised moderate to moderately severe patients over a period of 24 weeks, but has failed to demonstrate efficacy in mild to moderate AD and in institutionalised patients with severe disease. Galantamine has been shown to delay the onset of BPSD in mild to moderate AD patients in one placebo-controlled study, and improve BPSD in a similar study of patients with cerebrovascular disease or probable vascular dementia. Studies with rivastigmine have shown efficacy in placebo-controlled studies of mild to moderately severe AD and in patients with Lewy body variant AD. Institutionalised patients with severe disease also show symptomatic benefits in BPSD with rivastigmine, resulting in a reduction in concomitant psychoactive medication use. Symptom complexes responding to ChE inhibitors appear to differ - all agents improve apathy, depression and anxiety, while rivastigmine additionally improves hallucinations and delusions, possibility as a result of dual inhibition of acetylcholinesterase and butyrylcholinesterase. The presence of hallucinations has been shown to predict response to rivastigmine. Accumulating data from studies of ChE inhibitors suggest that early intervention and long-term treatment, in addition to providing cognitive benefits, improves BPSD and offers potential to enhance quality of life. Differences seen between the agents in terms of efficacy in BPSD, tolerability and safety profiles may be the result of differences in neuropharmacological profiles. PMID- 12094827 TI - Beneficial effects of L-carnitine in dialysis patients with impaired left ventricular function: an observational study. AB - BACKGROUND: Recent studies have shown that L-carnitine may improve clinical status and reduce the need for erythropoietin in dialysis patients with cardiovascular diseases. In this observational study, we investigated whether the addition of L-carnitine to conventional therapy might improve cardiac function (as assessed by M-mode and two-dimensional echocardiography) and clinical status in dialysis patients with left ventricular dysfunction. METHODS: Eleven dialysis patients with reduced left ventricular function (EF < 45%) were treated with L carnitine for 8 months. Two-dimensional (2-D) echocardiography was performed at baseline and every 2 months up to the end of the treatment period. The dosage of erythropoietin was also monitored during the study and the patients' clinical status was assessed by a questionnaire. RESULTS: Carnitine increased mean LV ejection fraction from 32.0% to 41.8% (p < 0.05 vs baseline). There was also a slight reduction of erythropoietin dosage and an improvement of clinical status. CONCLUSIONS: Eight months' therapy with carnitine appears to improve LV function and clinical status in dialysis patients with impaired LVF. PMID- 12094828 TI - Latvian community nurses practising in a time of turmoil: a thin line of defence for children at risk. AB - Recent World Health Organization (WHO) statistics rank Latvia below average on a number of health indicators, compared to other European Union (EU) candidate countries. Nurses who visit children in the home see a close-up view of the decline in health. This exploratory study was designed to understand the day-to day experience of a representative sample of 17 Latvian nurses visiting infants and children in 1999, almost 10 years after the collapse of the Soviet Union. Analysis of practice reports from group and individual interviews, using interpretive phenomenologic methods, revealed three consistently emerging themes: the negative impact of societal change and dislocation on clients and nurses; clinical reasoning concerning degree of risk to children and priorities for intervention; and the physical and emotional burden of caring for increasingly vulnerable children. Risk assessment was expressed in the classification of families as 'good little family, a family that was 'simply poor, or a family that was a 'social risk.' With the first two types of families, concern about children was balanced with concern about the family as a whole. With 'social risk' families, the primary concern of the nurses was safeguarding the child. The findings cannot be generalized, but may imbue family health and welfare statistics with a human face, and suggest directions for education, practice and further research. PMID- 12094829 TI - Critiquing bachelor candidates' theses: are the criteria useful? AB - Nursing education programmes should be at an academic level and connected to research. In Sweden, empirical studies are generally required in order to obtain a Bachelor's degree; hence, in some cases, these studies are replaced by a literature review. A study was conducted using 11 criteria. Thirteen theses produced in a department of nursing science were examined, elaborated and reproduced by reviewing international and national literature. Thereafter, the criteria themselves were scrutinized. Principal findings when critiquing the theses were that in eight theses the purpose was dearly identified and well defined in relation to the study accomplished; in three theses the purpose was indistinct and vague; and in two the definitions and research questions were lacking. The topic was relevant for the area of nursing in all theses. General problems identified were poor spelling and grammar, and unsatisfactory thesis structure. This article discusses whether criteria are useful when examining the Bachelor candidates' theses. The authors report that the criteria seemed to be useful, giving some guidance for scrutinizing theses and facilitating correspondence. Criteria could be appropriate guidelines for using to increase the quality of the theses as well as the quality of nursing. PMID- 12094830 TI - What nurses can do to empower young girls. PMID- 12094831 TI - ICN develops principles for ethical recruitment of nurses. PMID- 12094832 TI - INR authors speak out: the effect of war on children. PMID- 12094833 TI - ICN nurse consultant Fadwa Affara retires. PMID- 12094834 TI - Nurses and the tobacco epidenic. PMID- 12094835 TI - Culturally competent care: emphasis on understanding the people of Afghanistan, Afghanistan Americans, and Islamic culture and religion. AB - Since the attacks in New York and Washington, DC, in September 2001, increased racial and religious animosity has left Arabs, other Middle Easterners, Muslims, and those who bear physical resemblance to members of these groups, fearful. This article provides information about the people of Afghanistan, Afghanistan Americans, and Islamic culture and religion, which can greatly assist the nurse who is confronted with persons from diverse cultures during the provision of care. The Giger & Davidhizar Transcultural Assessment Model was first published in the International Nursing Review in 1990. This model is now used worldwide and provides an assessment model to assist in understanding cultural phenomena and individuals from different cultures. PMID- 12094836 TI - The effect of war on children: the children of Europe after World War II. AB - In war, children are inevitably innocent victims. In the carnage that was World War II, more children were killed or orphaned than at any other time in history. This article gives a brief history of the place of children within the conflagration, then describes the effects of war on the children. We concentrate on postwar life, placing children in the context of the environment in which they were living at the time. Our article outlines the work carried out by relief agencies and how Europe began to rebuild itself, how the children were fed and made healthy, and how, where possible, they were reunited with their families. We report briefly on the physical and psychological damage children suffered, both during the war and in its aftermath. History such as this is relevant to nurses in the 21st century, as it provides insight upon which nursing care for both our present ageing population and for children of the future can be based. PMID- 12094837 TI - Cross-mapping the ICNP with NANDA, HHCC, Omaha System and NIC for unified nursing language system development. International Classification for Nursing Practice. International Council of Nurses. North American Nursing Diagnosis Association. Home Health Care Classification. Nursing Interventions Classification. AB - Nursing language plays an important role in describing and defining nursing phenomena and nursing actions. There are numerous vocabularies describing nursing diagnoses, interventions and outcomes in nursing. However, the lack of a standardized unified nursing language is considered a problem for further development of the discipline of nursing. In an effort to unify the nursing languages, the International Council of Nurses (ICN) has proposed the International Classification for Nursing Practice (ICNP) as a unified nursing language system. The purpose of this study was to evaluate the inclusiveness and expressiveness of the ICNP terms by cross-mapping them with the existing nursing terminologies, specifically the North American Nursing Diagnosis Association (NANDA) taxonomy I, the Omaha System, the Home Health Care Classification (HHCC) and the Nursing Interventions Classification (NIC). Nine hundred and seventy-four terms from these four classifications were cross-mapped with the ICNP terms. This was performed in accordance with the Guidelines for Composing a Nursing Diagnosis and Guidelines for Composing a Nursing Intervention, which were suggested by the ICNP development team. An expert group verified the results. The ICNP Phenomena Classification described 87.5% of the NANDA diagnoses, 89.7% of the HHCC diagnoses and 72.7% of the Omaha System problem classification scheme. The ICNP Action Classification described 79.4% of the NIC interventions, 80.6% of the HHCC interventions and 71.4% of the Omaha System intervention scheme. The results of this study suggest that the ICNP has a sound starting structure for a unified nursing language system and can be used to describe most of the existing terminologies. Recommendations for the addition of terms to the ICNP are provided. PMID- 12094838 TI - What we need is a reliable plug. PMID- 12094839 TI - Equimolar doses of balsalazide and mesalamine: are we comparing apples and oranges? PMID- 12094840 TI - Liver biopsy: who should do it...and who will show up in court? PMID- 12094841 TI - Hepatobiliary scintigraphy as the diagnostic test for sphincter of Oddi dysfunction: pan the scan? PMID- 12094842 TI - Quality in the technical performance of colonoscopy and the continuous quality improvement process for colonoscopy: recommendations of the U.S. Multi-Society Task Force on Colorectal Cancer. PMID- 12094843 TI - A critical evaluation of laboratory tests in acute pancreatitis. AB - An ideal laboratory test in the evaluation of a patient with acute pancreatitis (AP) should, in addition to accurately establishing the diagnosis of AP, provide early assessment of its severity and identify the etiology. None of the tests available today meet all these criteria, and presently there is no biochemical test that can be considered the "gold standard" for the diagnosis and assessment of severity of AP. In the diagnosis of AP, serum amylase and lipase remain important tests. Advantages of amylase estimation are its technical simplicity, easy availability, and high sensitivity. However, its greatest disadvantage is its low specificity. A normal amylase would usually exclude the diagnosis of AP, with the exception of AP secondary to hyperlipidemia, acute exacerbation of chronic pancreatitis, and when the estimation of amylase is delayed in the course of the disease. The major advantage of lipase is an increased sensitivity in acute alcoholic pancreatitis and in patients who initially present to the emergency room days after the onset of the disease, as lipase remains elevated longer than amylase. Although once considered to be specific for AP, nonspecific elevations of lipase have been reported in almost as many disorders as amylase, thus decreasing its specificity. Simultaneous estimation of amylase and lipase does not improve the accuracy. Other enzymes for the diagnosis of AP--pancreatic isoamylase, immunoreactive trypsin, and elastase--are more cumbersome and expensive and have no clear role in the diagnosis of AP. No enzyme assay has a predictive role in determining the severity or etiology of AP. Once the diagnosis of AP is established, daily measurements of enzymes have no value in assessing the clinical progress of the patient or ultimate prognosis and should be discouraged. A host of new serological and urinary markers have been investigated in the last few years. Their main use is in predicting the severity of AP. At present, serum C-reactive protein at 48 h is the best available laboratory marker of severity. Urinary trypsinogen activation peptides within 12-24 h of onset of AP are able to predict the severity but are not widely available. Serum interleukins 6 and 8 seem promising but remain experimental. PMID- 12094844 TI - Upper endoscopy as a screening and surveillance tool in esophageal adenocarcinoma: a review of the evidence. AB - Esophageal adenocarcinoma is a rare cancer that is increasing rapidly in incidence. Because gastroesophageal reflux disease (GERD) is a risk factor for the development of this cancer, endoscopic screening of individuals with GERD symptoms and endoscopic surveillance of those who are found to have Barrett's esophagus (BE), the presumed precursor to adenocarcinoma, have been proposed. Although no direct data support endoscopic screening or surveillance, several lines of indirect evidence are available. We apply a set of criteria for the evaluation of screening programs to endoscopic screening of subjects with reflux and endoscopic surveillance of subjects with BE. A critical examination of the data supporting these practices shows that considerable gaps exist in our knowledge regarding endoscopy as a screening test in GERD, making us unable to support this practice based on current evidence. Although no controlled trials exist to substantiate the effectiveness of surveillance programs for subjects with BE, some stronger indirect evidence does support this practice. However, further studies are necessary to substantiate the effectiveness and cost effectiveness of endoscopic surveillance in BE. Based on the currently available data, consideration should be given to expanding the intervals between endoscopic surveillance sessions. PMID- 12094845 TI - Lifestyle factors and Barrett's esophagus. AB - OBJECTIVE: We aimed to investigate lifestyle factors relevant to the development of Barrett's esophagus in the United Kingdom. METHODS: At Ninewells Hospital, Dundee, Scotland, medical records of 136 Barrett's esophagus patients were examined. At Wexham Park Hospital, Slough, Southern England, 50 male and 51 female Barrett's esophagus patients were matched for sex, age, and year of diagnosis (+/- 3 yr) with uncomplicated reflux esophagitis patients. Data were abstracted for tobacco consumption, alcohol intake, and weight. In Dundee, height was also recorded and body mass index calculated. Alcohol and tobacco intake were scored for each patient. RESULTS: In Dundee there is no difference in smoking or drinking habits between men and women under and over 50 yr of age. In Slough there is little difference in drinking or smoking habits between Barrett's esophagus and reflux esophagitis patients and between their mean weights. However, of the Dundee Barrett's esophagus patients younger than 50 yr, 31% of men and 71% of women have body mass indexes over 30 (obese), versus 11% and 13%, respectively, for the general population. In those older than 50 yr, 14% of men and 19% of women have body mass indexes over 30. CONCLUSIONS: There is no difference in smoking or drinking habits in younger and older Barrett's esophagus patients, nor between those with Barrett's esophagus and reflux esophagitis. Obesity is a risk factor for Barrett's esophagus in young people only. PMID- 12094846 TI - Rabeprazole in nonerosive gastroesophageal reflux disease: a randomized placebo controlled trial. AB - OBJECTIVES: Clinical results to date suggest that antisecretory therapy may be less effective in providing symptom relief for patients with nonerosive gastroesophageal reflux disease (GERD) than for patients with erosive disease. This study was carried out to assess the efficacy and rapidity of once-daily rabeprazole (10 mg or 20 mg) in relieving symptoms in endoscopically negative patients with moderately severe GERD symptoms and to evaluate the safety of these doses over 4 wk. METHODS: This placebo-controlled, double blind study enrolled 203 men and women with moderately severe symptoms of GERD. After a 2-wk, single blind placebo run-in phase, patients were randomized to receive 10 mg or 20 mg of rabeprazole or placebo once daily for 4 wk. RESULTS: Rabeprazole rapidly and effectively relieved heartburn, with significant improvements on day 1 of dosing. It also improved most other GERD-related symptoms, including regurgitation, belching, bloating, early satiety, and nausea. Both rabeprazole doses were significantly superior to the placebo with respect to time to the first 24-h heartburn-free interval (2.5 and 4.5 days for 10 mg and 20 mg of rabeprazole, respectively, vs 21.5 days for the placebo) and first daytime or nighttime heartburn-free interval (1.5-3 days for rabeprazole groups vs 12.5-15 days for the placebo), as well as to percentage of time patients were heartburn-free and free of antacid use. Both rabeprazole doses were well tolerated. CONCLUSIONS: Based on these findings and prior studies, rabeprazole reliably relieves GI symptoms equally well in both nonerosive GERD and erosive GERD. PMID- 12094848 TI - Long term results of pneumatic dilation in achalasia followed for more than 5 years. AB - OBJECTIVE: We aimed to evaluate the long term therapeutic outcome in achalasia patients treated with pneumatic dilation, specifically focusing on those patients treated more than 15 yr ago. METHODS: All patients treated in our center whose records were available for review were asked to fill out a questionnaire assessing their degree of dysphagia, retrosternal pain, regurgitation, weight loss, and coughing during the night. The number of dilations was collected from the clinical records. The results of the treatment were classified into four different classes (excellent, good, moderate, poor). For those patients who had died, the cause of death was ascertained from the medical records or from the general practitioner. RESULTS: The questionnaires were distributed to 249 patients, 32 of whom had died. Of the 125 patients who completed the questionnaire, 81 (45 male and 36 female) were treated more than 5 yr ago. The mean follow-up was 12+/-1 yr. The therapeutic success rate was 50%, obtained after a median of four dilations (interquartile range = 3-6). Of this cohort, 25 patients (18 male and seven female, aged 35-84 yr) were treated more than 15 yr ago (mean follow-up = 20.5+/-0.5 yr). The median number of dilations was four (interquartile range = 3-7), with a therapeutic success rate of 40%. Two patients experienced a perforation, and seven were referred for surgery. Six patients out of 32 (19%) died of esophageal cancer. CONCLUSIONS: The long term success rate of pneumatic dilation is rather low, resulting in permanent successful treatment of achalasia in only 40-50% of patients. Achalasia is a risk factor for esophageal cancer. PMID- 12094847 TI - Acid suppression therapy may not alter malignant progression in Barrett's metaplasia showing p53 protein accumulation. AB - OBJECTIVES: Several previous studies have shown that malignant progression in Barrett's metaplasia (BM) occurs even in patients treated with fundoplication or acid suppression therapy (AST). The aim of this study was to test the hypothesis that AST may not alter malignant progression in BM if key genes involved in DNA repair and cell cycle control, particularly p53, are defective. METHODS: Initial and follow-up biopsies from 21 patients with BM treated with AST and observed for 1-13 yr were entered in the study. All biopsies were graded for dysplasia and evaluated for p53 protein accumulation and oxidative DNA damage by immunohiostochemistry, using antibodies to p53 and to 8-hydroxydeoxyguanosine, respectively. DNA ploidy was determined using image analysis. Statistical analysis was performed using Kaplan-Meier curves, log rank test, and multivariate regression. RESULTS: Patients with p53 positive initial biopsies were more likely to have progression in dysplasia grade (p = 0.022) and DNA ploidy status (p = 0.023) than those with p53 negative biopsies. In eight patients AST resulted in significant reduction in oxidative DNA damage in the five patients with p53 negative initial biopsies, but not the three with p53 positive ones (p = 0.0007). CONCLUSIONS: We conclude that failure of AST to alter malignant progression in BM may be due, at least in part, to defects in DNA repair and cell cycle control resulting from p53 gene mutation, present before AST treatment. Although AST may be effective in preventing further DNA damage, it is unlikely to alter progression in genetically unstable cells. PMID- 12094849 TI - Gastroesophageal reflux disease in chronic renal failure patients with upper GI symptoms: multivariate analysis of pathogenetic factors. AB - OBJECTIVE: The association between gastroesophageal reflux disease and end-stage renal disease remains unclear. We aimed to assess the prevalence of gastroesophageal reflux disease and also to identify possible pathogenetic factors in the development of reflux in symptomatic end-stage renal disease patients. METHODS: The study involved 42 end-stage renal disease patients with upper GI symptoms (group I) and 46 age- and sex-matched controls who did not have renal disease but had the same symptoms (group II). Endoscopy, endoscopic biopsies, and 24-h esophageal pH studies were used to diagnose gastroesophageal reflux disease. Subjects were also investigated for Helicobacter pylori gastritis and GI amyloidosis. RESULTS: The prevalences of gastroesophageal reflux disease in the two groups were similar (81% vs 84.8%, p = 0.423). The prevalence of H. pylori infection was significantly lower in group I than in group II (38.1% vs 67.4%, p = 0.01). There were II cases of GI amyloidosis in group I. Multivariate logistic regression analysis in group I showed that GI amyloidosis (OR = 7.28, 95% CI = 1.13-46.93), chronic ambulatory peritoneal dialysis treatment (OR = 5.54, 95% CI = 1.01-30.43), and absence of H. pylori infection (OR = 3.75, 95% CI = 1.01-13.9) were significantly associated with reflux esophagitis. CONCLUSIONS: Upper GI symptoms are important in predicting gastroesophageal reflux disease in end-stage renal disease patients. Chronic ambulatory peritoneal dialysis, GI amyloidosis, and absence of H. pylori infection seem to be risk factors for the development of gastroesophageal reflux disease in end-stage renal disease patients. PMID- 12094850 TI - Daily low-dose versus alternate day full-dose lansoprazole in the maintenance treatment of reflux esophagitis. AB - OBJECTIVE: For the long term maintenance treatment of reflux esophagitis several strategies have been proposed with the aim of reducing the daily dosage or the frequency of drug administration. However, the available clinical studies are scarce and are often not controlled or conducted on a reduced number of cases. We aimed to compare the efficacy of two doses of lansoprazole (15 mg once daily and 30 mg on alternate days) in maintaining endoscopic healing and symptom relief over a 6-month period. METHODS: One hundred thirty-seven patients with Savary Miller grades I-III reflux esophagitis healed after an 8-wk treatment with lansoprazole (30 mg daily) were divided into two main groups for a 6-month maintenance therapy period: lansoprazole, 15 mg once daily (group 15qd) and lansoprazole, 30 mg on alternate days (group 30qod). These two main groups were further subdivided according to the time of drug administration; morning (15qdm and 30qodm) and evening (15qde and 30qode). Each patient underwent esophagogastroduodenoscopy before entry into the study, after 8 wk of acute therapy, and after 6 months of maintenance therapy; 24-h esophageal-gastric pH monitoring was performed at baseline and during the last week of maintenance therapy. RESULTS: At the end of the maintenance period the recurrence of esophagitis was observed in 12.1% of group 15qd patients and in 19.0% of group 30qod patients, without significant differences between the two groups. The frequency of patients without reflux symptoms after the 6-month period was the same for both groups; however, a significant increase of heartburn was observed in group 30qod patients (from 12.1% to 28.6%, p = 0.007). The time of drug administration (morning and evening) had no influence on the outcome of treatment. Both regimens significantly reduced esophageal acid exposure time and increased the median 24-h gastric pH. CONCLUSIONS: Both long term lansoprazole regimens are equally effective in preventing the recurrence of esophagitis, independent of the modality of drug administration. The daily administration seems to have a better effect on the prevention of symptom recurrence. PMID- 12094851 TI - Histological patterns of gastritis in H. pylori-infected individuals with a family history of gastric cancer. AB - OBJECTIVE: Different types of chronic gastritis, including antral predominant, corpus predominant, and multifocal pangastritis, are associated with Helicobacter pylori infection. Specific patterns of H. pylori gastritis that might characterize individuals with family histories of noncardia gastric cancer (GC) were investigated. METHODS: Histopathological changes associated with H. pylori gastritis were assessed in 111 individuals with family histories of GC and in 77 without from a region with high prevalence of H. pylori infection and GC. Gastric biopsies were taken from 12 sites (antrum, five; corpus, six; and cardia, one). RESULTS: Individuals (age < 36 yr) with family histories of GC developed pangastritis and had higher H. pylori bacterial scores (p < 0.05) in the gastric corpus, whereas those without family histories of GC typically had antral predominant gastritis. The correlation between density of polymorphonuclear leukocytes and density of H. pylori at each biopsy site was statistically significant (p < 0.01). Pangastritis was associated with a higher density of lymphoid aggregates and follicles (p < 0.05) in the corpus of younger individuals (age < 36) and in the antrum of older individuals (age > or = 48) with positive family histories of GC. CONCLUSIONS: Pangastritis and high lymphoid follicle density associated with H. pylori infection were found in patients with family histories of GC. Because a family history of gastric carcinoma is associated with increased risk of gastric cancer development, characterization of histological patterns of gastritis may be applicable to gastric cancer screening and surveillance, especially in relatively young at-risk populations. PMID- 12094852 TI - Role of Helicobacter pylori infection in the incidence and clinical course of monoclonal gammopathy of undetermined significance. AB - OBJECTIVES: We aimed to evaluate the hypothesis that chronic antigenic stimulation is an etiological factor of monoclonal gammopathy of undetermined significance (MGUS) with special relevance to Helicobacter pylori infection. METHODS: We performed a retrospective, observational study. The diagnosis of MGUS was based on serum protein electrophoresis, serum IgM levels (<3 gr/dl), urinary M protein, a normal bone marrow biopsy, normal renal function, and a negative skeletal survey. H. pylori infection was ascertained by rapid urease testing and endoscopy with gastric biopsy for histology. Eradication of H. pylori was assessed by urea breath test and stool antigen assay 4 wk after completion of an appropriate treatment regimen. RESULTS: Sixty-nine patients with MGUS were included in the study. Of these, 57 had undergone evaluation for H. pylori infection for various GI symptoms. Thirty-nine of 57 patients (68.42%) with MGUS also had evidence of H. pylori infection. In 11 of these 39 patients (28.21%), eradication of H. pylori infection with an appropriate regimen led to normalization of the serum protein electrophoresis and resolution of the gammopathy. CONCLUSION: The results of our study give increased credibility to the theory that in a proportion of patients the pathogenesis of MGUS involves chronic antigenic stimulation and H. pylori is implicated. The search for H. pylori infection and an attempt to eradicate the bacterium in positive cases seem to be appropriate in patients diagnosed with MGUS. PMID- 12094853 TI - Small adenocarcinomas of the esophagogastric junction: association with intestinal metaplasia and dysplasia. AB - OBJECTIVES: Intestinal metaplasia in Barrett's esophagus predisposes to esophageal adenocarcinoma. Intestinal metaplasia of the cardia is a common finding in persons without cancer. Many adenocarcinomas of the esophagogastric junction are large enough to obliterate any underlying intestinal metaplasia. To estimate how often adenocarcinoma of the esophagogastric junction arises in intestinal metaplasia, we studied small adenocarcinomas of the esophagogastric junction. METHODS: Resection patients had adenocarcinomas 2 cm or smaller, within 2 cm of the esophagogastric junction. Age- and sex-matched controls had resection for squamous carcinoma. Saved and new histological slides from the esophagogastric junction were examined, with additional stains. RESULTS: Intestinal metaplasia was found in 86% (19/22) of adenocarcinoma cases, versus 32% (7/22) of controls (p < 0.001). Intestinal metaplasia with high or low grade dysplasia was associated with 64% (14/22) of adenocarcinomas and with 5% (1/22) of controls (p < 0.001). Excluding four cases with long and three with short Barrett's esophagus, 80% (12/15) of adenocarcinomas had associated intestinal metaplasia, 53% (8/15) with dysplasia. Most adenocarcinoma cases had the incomplete type of intestinal metaplasia with a Barrett type cytokeratin 7/20 staining pattern. Helicobacter pylori were seen in one adenocarcinoma and five control cases. CONCLUSIONS: Most adenocarcinomas of the esophagogastic junction arise in the background of intestinal metaplasia, sometimes in an endoscopically visible Barrett's esophagus, more often in small areas of intestinal metaplasia of the cardia. In cases of adenocarcinoma, the intestinal metaplasia resembled that found in Barrett's esophagus, and was not associated with H. pylori. PMID- 12094854 TI - Endoscopic use of human thrombin in bleeding gastric varices. AB - OBJECTIVE: The management of gastric variceal hemorrhage remains a clinical challenge. Bovine thrombin has been reported to be effective in two small series. We report our experience with human thrombin in the treatment of bleeding gastric varices. METHODS: We reviewed the case records of 12 patients presenting over a 2 yr period with gastric variceal bleeding requiring endoscopic injection of human thrombin. Ten were male and the mean age was 52 yr (range = 26-83). The underlying diagnoses were cirrhosis in nine, portal vein thrombosis in two, and liver metastasis in one. The majority had fundal gastric varices, and none were thought to have bled from their esophageal varices. Eight received thrombin as primary treatment, whereas four had thrombin only after failing transjugular intrahepatic portosystemic shunts. Patients received one to four sessions (mean = 1.9) of thrombin with a mean total dose of 1833 U (range = 800-4000). Mean follow up was 17.8 months for those still alive (range = 7-33). RESULTS: Hemostasis in the acute setting was successful in nine patients all of whom received thrombin within 48 hours of the bleed. In the longer term, nine of the 12 had no further bleeding. Of these, five patients did well with thrombin alone, one died of cancer, and the other three went on electively to more definitive shunt procedures. Three patients rebled from their gastric varices of which one was successfully retreated with thrombin. Only one death was related to variceal bleeding (8%). No adverse reactions were noted. CONCLUSION: Our experience demonstrates that endoscopic therapy with thrombin appears safe and can be effective in the management of gastric variceal bleeding. PMID- 12094855 TI - Endosonography-guided fine needle aspiration biopsy in the evaluation of pancreatic masses. AB - OBJECTIVES: Diagnosis of pancreatic tumors can be problematic. This study aimed to determine the performance of endoscopic ultrasound-guided fine needle aspiration biopsy (EUS FNA) in pancreatic malignancy when prior biopsies performed by CT guidance or ERCP were negative. METHODS: A total of 185 patients with known or suspected pancreatic masses were prospectively evaluated with EUS FNA. Before EUS FNA, all patients were evaluated with abdominal CT (61 with CT guided biopsy) and 91 with ERCP (41 had brushings or biopsy). RESULTS: EUS had greater sensitivity than CT in detecting a mass (99% vs 57%, p < 0.0001). In 58 patients with negative CT-guided biopsies, EUS FNA had 90% sensitivity for malignancy, 50% specificity for benign disease and 84% accuracy. Similarly, in 36 patients with negative ERCP tissue sampling, results for EUS FNA were 94%, 67% and 92%, respectively. Complications were mild and infrequent (0.5%). CONCLUSION: EUS FNA of pancreatic masses safely and accurately diagnoses pancreatic malignancy when prior biopsy techniques have been unsuccessful. PMID- 12094856 TI - Cholecystokinin-stimulated peak lipase concentration in duodenal drainage fluid: a new pancreatic function test. AB - OBJECTIVE: Hormonal stimulation with secretin or cholecystokinin (CCK) is the most sensitive means of assessing pancreatic function. Secretin is not available, and current CCK tests are cumbersome, requiring dual tube intubation and marker perfusion techniques. The aim of this study was to test the efficacy of a new CCK stimulated pancreatic function test measuring peak lipase concentration. METHODS: A Dreiling gastroduodenal tube was inserted to the ligament of Treitz, and fluid was collected on ice for 80 min in four 20-min aliquots. CCK was infused i.v. at a constant rate of 40 ng/kg/h. Gastric aspirations were discarded. Duodenal aspirates were analyzed for volume and enzyme concentration with a clinical laboratory autoanalyzer. RESULTS: Nineteen healthy volunteers and 18 chronic pancreatitis patients were studied. Lipase concentration and secretory volume showed a peak response by 40 min of stimulation, whereas amylase response was variable. The mean peak lipase concentrations (+/-SEM) for normal volunteers and mild, moderate, and advanced chronic pancreatitis patients were 16.9+/-1.9, 7.9+/ 1.7, 3.7+/-1.2, and 2.1+/-0.6 x 10 5 IU/L, respectively. Lower peak lipase concentrations were significantly associated with more advanced chronic pancreatitis (p < 0.001). The receiver operating characteristic curve area for all chronic pancreatitis patients was 0.944 (95% CI = 0.825-0.985). A peak lipase concentration of 780,000 IU/L provided a sensitivity and specificity of 0.833 and 0.867, respectively. This CCK test was well tolerated and without complications. CONCLUSIONS: Lipase concentration in duodenal fluid increases nearly 3-fold from baseline after CCK stimulation in healthy volunteers but is markedly reduced in patients with chronic pancreatic disease. Peak lipase concentration is a significant predictor of chronic pancreatitis and correlates with severity of pancreatic disease. Aspiration of duodenal drainage fluid with a Dreiling tube and analysis with a laboratory autoanalyzer are less cumbersome than marker perfusion and back titration techniques. Measurement of enzyme concentration instead of output could lead to the development of an endoscopic or through-the scope screening method for assessing patients with suspected chronic pancreatitis or chronic abdominal pain. PMID- 12094857 TI - A randomized, double blind, dose-response comparison of balsalazide (6.75 g), balsalazide (2.25 g), and mesalamine (2.4 g) in the treatment of active, mild-to moderate ulcerative colitis. AB - OBJECTIVE: Balsalazide is a new innovative, mesalamine-containing prodrug that is activated by bacteria in the colon. Balsalazide has been shown previously to be well tolerated and effective in the treatment of acute ulcerative colitis. The aim of this study was to determine the dose-response of balsalazide for efficacy and safety in active, mild-to-moderate ulcerative colitis and to compare this profile with that of mesalamine, pH-dependent, delayed-release tablets. METHODS: A multicenter, randomized, active control, double-blind, double-dummy, dose response, parallel-group study was performed comparing balsalazide (6.75 g daily), balsalazide (2.25 g daily), and mesalamine (2.4 g daily), administered for 8 wk to 154 patients with active, mild-to-moderate ulcerative colitis as verified by sigmoidoscopy. RESULTS: Eight weeks of treatment with 6.75 g of balsalazide daily provided significantly greater improvement than did balsalazide (2.25 g daily) in rectal bleeding (64.7% [6.75-g balsalazide] vs 32.4% [2.25-g balsalazide], p < 0.006), stool frequency (58.8% vs 29.4%, p < 0.006), sigmoidoscopic score (78.9% vs 52.5%, p < 0.015), and Physician's Global Assessment (73.7% vs 51.3%, p < 0.03). The efficacy of balsalazide showed a significantly more rapid onset of action than that of mesalamine (2.4 g daily) (2 wk sigmoidocopic score improvement, 54.7% [6.75-g balsalazide] vs 29.4% [2.4-g mesalamine], p = 0.006) with numerically greater improvement at 8 wk in five of seven measured signs and symptoms. Balsalazide (6.75 g daily) was well tolerated, and the safety profile did not differ significantly from that of balsalazide (2.25 g daily) or mesalamine. CONCLUSIONS: Eight weeks of treatment with balsalazide (6.75 g daily) is significantly more effective than balsalazide (2.25 g daily) and more rapid in onset than mesalamine (2.4 g daily) in improving signs and symptoms of acute ulcerative colitis. Balsalazide (6.75 g daily) is well tolerated, and the safety profile does not differ from that of balsalazide (2.25 g daily) and mesalamine (2.4 g daily). PMID- 12094858 TI - Infliximab retreatment in adults and children with Crohn's disease: risk factors for the development of delayed severe systemic reaction. AB - OBJECTIVES: Although effective in the treatment of refractory Crohn's disease, episodic retreatment with the antitumor necrosis factor a chimeric monoclonal antibody infliximab (Remicade, Centocor, Malvern, PA) can be associated with severe acute and delayed systemic reactions. METHODS: We analyzed episodic infliximab retreatment over 30 months in 86 adult and pediatric patients receiving 304 infusions to determine factors associated with the development of severe systemic reaction. RESULTS: Overall, 14% of patients experienced severe systemic reactions with episodic infliximab retreatment. There was a significant difference in the rates of severe systemic reaction observed in adults (11/52 [21%]) and pediatric patients (1/34 [3%]) (p < 0.02). Delayed systemic reactions, characterized by arthralgia, fever, and myalgia requiring corticosteroid treatment, were found exclusively in adults (age > 17 yr) and occurred in eight patients treated for luminal Crohn's disease. Acute systemic reactions, characterized by hypotension, mucosal irritability, and laryngospasm requiring epinephrine, diphenhydramine, and/or methylprednisolone treatment, occurred sporadically in three adults and one child, treated for both luminal and fistulizing disease. Second infliximab infusions were associated with two thirds of severe systemic reactions, and a distant second infusion (> or = 20 wk from first infusion) was poorly tolerated relative to earlier retreatment (p < 0.001). Concomitant medications were similar in adults and children. CONCLUSIONS: Episodic infliximab retreatment--specifically, a distant second infusion--is associated with high rates of severe systemic reaction in adults, but not children. We recommend multiple early infusions of infliximab if retreatment is anticipated in adult patients to avoid the development of delayed severe systemic reactions. PMID- 12094859 TI - Focally enhanced gastritis in children with Crohn's disease and ulcerative colitis. AB - OBJECTIVES: Focally enhanced gastritis (FEG) has been suggested as a specific diagnostic marker for patients with Crohn's disease. However, the utility of FEG for distinguishing Crohn's disease from ulcerative colitis is uncertain in adults, and the occurrence of this lesion in children has not been defined. The aim of this study was to evaluate the occurrence of FEG and other gastric histological abnormalities in children with inflammatory bowel disease (IBD) and to examine the utility of FEG in discriminating between ulcerative colitis and Crohn's disease. METHODS: This is a retrospective, case-controlled study of upper GI histopathological findings in children with IBD. Gastric histopathology was defined and graded according to the Updated Sydney System. RESULTS: FEG was present in 28 of 43 (65.1%) children with Crohn's disease and five of 24 (20.8%) children with ulcerative colitis, compared to three of 132 (2.3%) children without IBD or one of 39 (2.6%) children with Helicobacter pylori infection. There were no differences between those with and without FEG with regard to upper GI symptoms or previous anti-inflammatory drug ingestion (5-aminosalicylic acid compounds or steroids). All patients with H. pylori infection had chronic antral gastritis, but only one child with H. pylori had FEG. In addition, mild to moderate chronic gastritis was present in 15 of 43 (34.9%) children with Crohn's disease and in 12 of 24 (50%) patients with ulcerative colitis. CONCLUSIONS: The presence of FEG suggests underlying IBD. Although FEG is particularly common in children with Crohn's disease, it does not reliably differentiate between Crohn's disease and ulcerative colitis. PMID- 12094860 TI - Induction of apoptosis before shedding of human intestinal epithelial cells. AB - OBJECTIVES: Human intestinal epithelial cells (IECs) derive from stem cells at the crypt base and migrate along the so-called crypt-villus axis toward the intestinal lumen. As they reach the luminal surface in the colon or the villus tip in the small intestine, IECs are shed and their life cycle is terminated. The role of apoptosis during IEC migration along the crypt-villus axis has been subject to studies with conflicting results. In this study we use a novel approach to identify the initiation of apoptosis within normal human IECs. METHODS: Normal mucosa from the large and small human intestine was analyzed employing a novel antibody directed against activated caspase-3--an early marker of apoptosis. RESULTS: IECs initiate the apoptotic cascade as they approach the area of shedding before displaying evident morphological features of apoptosis. IECs of the small bowel also show caspase-3 activation in the small intestinal crypt base, whereas IECs of the colononic crypt base rarely show evidence of ongoing apoptosis. CONCLUSIONS: These findings indicate that apoptosis is initiated in human IECs as they reach the luminal surface/villus tip and before shedding. Furthermore, they show that different sections of the intestinal tract vary significantly in the rate of IEC apoptosis, possibly reflecting their difference in susceptibility to epithelial cell neoplasia. PMID- 12094861 TI - Assessing the quality of life of household members of patients with inflammatory bowel disease: development and validation of a specific questionnaire. AB - OBJECTIVE: Household members of patients with inflammatory bowel disease (IBD) are impacted in their daily lives by close interaction with chronic illness. However, the effect of IBD on the quality of life of household members has not been evaluated because appropriate instruments to measure it are lacking. The aim of this study was to develop and validate a specific instrument to measure the quality of life of household members of patients with IBD. METHODS: Household members were interviewed to generate an initial pool of items. A group of investigators excluded redundant and incomprehensible items, and the resulting preliminary questionnaire was completed by 81 subjects. Further item reduction was performed using the psychometric method. The psychometric properties of the final questionnaire were determined in a group of 106 subjects. RESULTS: Of the initial 67 items, 19 were excluded by investigators. The remaining 48 items were answered by 81 household members, and 32 items were excluded by factor analysis. Two others were excluded because of poor comprehension. The definitive questionnaire consisted of 14 items. CONCLUSIONS: We have developed and validated a questionnaire to measure the quality of life of household members of IBD patients. It will greatly enhance the possibilities of quantifying the effects of IBD on home life and should become a useful tool in health care planning. PMID- 12094863 TI - Differentiation between intestinal tuberculosis and Crohn's disease in endoscopic biopsy specimens by polymerase chain reaction. AB - OBJECTIVES: It is difficult to differentiate intestinal tuberculosis from Crohn's disease because of similar clinical, pathological, radiological, and endoscopic findings. The purpose of this study was to investigate the value of polymerase chain reaction (PCR) assay in the differentiation intestinal tuberculosis from Crohn's disease, and compare the histopathological features of endoscopic biopsy of the two disorders. METHODS: A total of 39 endoscopic biopsy specimens from patients with intestinal tuberculosis and 30 specimens from patients with Crohn's disease were subjected to pathological analysis retrospectively, Ziehl-Neelsen stain, and PCR assay. RESULTS: Except for granuloma with caseation and confluence, which was the characteristic of intestinal tuberculosis, other pathological features of intestinal tuberculosis and Crohn's disease were very similar or were difficult to find in endoscopic biopsy specimens. The positivity rate by PCR in 39 intestinal tuberculosis specimens was 64.1% (25/39), but was zero by PCR in 30 Crohn's disease specimens. Moreover, in the tissues of intestinal tuberculosis with granulomas similar to those of Crohn's disease, there were 71.4% (10/14) positive by PCR, and there were 61.1% (11/18) positive in intestinal tuberculosis tissues without granulomas. CONCLUSIONS: Biopsy is of limited diagnostic value in the differentiation intestinal tuberculosis from Crohn's disease, and PCR is valuable in the differentiation between intestinal tuberculosis and Crohn's disease. PMID- 12094864 TI - Elevated serum eotaxin levels in patients with inflammatory bowel disease. AB - OBJECTIVE: Eotaxin is a recently characterized chemokine with potent and selective chemotactic activity for eosinophils. Previous studies indicating that eosinophils accumulate and become activated in inflammatory bowel disease (IBD) led us to hypothesize that eotaxin is potentially involved in the pathophysiology of IBD and, therefore, that eotaxin would be increased in the serum of patients with IBD. The objective of this study was to test those assumptions. METHODS: We investigated 72 patients with IBD, 35 with ulcerative colitis, and 37 with Crohn's disease. A total of 27 patients had active and 45 inactive disease; 26 were receiving corticosteroids. Eotaxin serum levels were determined by solid phase sandwich ELISA. Lymphocytes, monocytes, and granulocyte subpopulations were determined in fresh blood samples with an automated autoanalyzer. RESULTS: Serum eotaxin levels were significantly higher in patients with Crohn's disease and in those with ulcerative colitis than in the control subjects (p < 0.0001). Patients with inactive Crohn's disease had significantly higher levels of eotaxin than patients with inactive ulcerative colitis (p < 0.05). We did not find significant differences for activity or inactivity of disease, nor for treatment with prednisone. A negative correlation (p < 0.05) was found between eotaxin serum level and eosinophil counts in peripheral blood in patients with Crohn's disease. CONCLUSIONS: There is an increased expression of eotaxin in IBD patients, suggesting that eotaxin may be involved in the pathogenesis of IBD. This increase is more accentuated in Crohn's disease and negatively correlates with the eosinophil number in peripheral blood. Our data support the increasing evidence that eosinophil are functionally involved in the pathophysiology of IBD. PMID- 12094862 TI - Prior bowel resections, perianal disease, and a high initial Crohn's disease activity index are associated with corticosteroid resistance in active Crohn's disease. AB - OBJECTIVES: Some patients with Crohn's disease (CD) do not respond to corticosteroid therapy. Furthermore, corticosteroids frequently cause side effects. Thus, predictive parameters for treatment refractoriness would be helpful for treatment decisions. METHODS: A total of 300 patients with active CD (i.e., with a Crohn's Disease Activity Index [CDAI] >200) entered the study. Treatment started with 60-100 mg/day prednisolone equivalent, which was then tapered to 10-15 mg/day within 6 wk and maintained at that dose for another 4 wk. After 10 wk of treatment, response to steroids was defined by a CDAI <150, steroid resistance by a CDAI always > or =150 and steroid dependency by a relapse after dose reduction. Of 239 eligible patients, 196 were responders, 26 were steroid resistant, and 17 were steroid dependent. RESULTS: Prior bowel resections, a high initial CDAI, and perianal disease were associated with steroid resistance. Of the steroid resistant patients 53.9% were bowel-resected compared to 20.4% of the responders (relative risk = 3.63; 95% CI = 1.79-7.36). Perianal disease was observed in 42.3% of steroid resistant patients versus 21.9% of responders (relative risk = 2.28; 95% CI = 1.12-4.66) and initial CDAI was 347+/-91 in resistant patients versus 301+/-81 in responders (p < 0.05). Parameters for steroid dependent patients were not significantly different from those of responders. CONCLUSIONS: In this study (thus far the largest study for the evaluation of predictive factors for treatment refractoriness to corticosteroids in CD), only prior bowel resection, perianal disease, and a high initial CDAI were found to be predictive of resistance to steroid treatment. PMID- 12094865 TI - Serological markers for prediction of response to anti-tumor necrosis factor treatment in Crohn's disease. AB - OBJECTIVES: The use of monoclonal anti-tumor necrosis factor (TNF) antibodies (infliximab, Remicade) is a new therapeutic approach for severe refractory luminal or fistulizing, Crohn's disease (CD). However, up to 30% of patients do not respond to this treatment. So far, no parameters predictive of response to anti-TNF have been identified. Our aim was to determine whether serological markers ASCA (anti-Saccharomyces cerevisiae antibodies) or pANCA (perinuclear antineutrophil cytoplasmic antibodies) could identify Crohn's patients likely to benefit from anti-TNF therapy. METHODS: Serum samples of 279 CD patients were analyzed for ASCA and pANCA before anti-TNF therapy. A blinded physician determined clinical response at week 4 (refractory luminal CD) or week 10 (fistulizing CD) after the first infusion of infliximab (5 mg/kg). RESULTS: Overall, there was no relationship between ASCA or pANCA and response to therapy. However, lower response rates were observed for patients with refractory intestinal disease carrying the pANCA+/ASCA- combination, although this lacked significance (p = 0.067). CONCLUSIONS: In this cohort of infliximab-treated patients, neither ASCA nor pANCA could predict response to treatment. However, the combination pANCA+/ASCA- might warrant further investigation for its value in predicting nonresponse in patients with refractory luminal disease. PMID- 12094866 TI - Celiac disease and irritable bowel-type symptoms. AB - OBJECTIVES: Previous reports have linked irritable bowel syndrome (IBS) etiologically with various forms of mucosal inflammation, including infectious enterocolitides and inflammatory bowel disease. The mechanism is uncertain but may involve sensitization by inflammatory mediators. The enteropathy of celiac disease has theoretical advantages as a study model because it can be controlled with dietary gluten exclusion; however, whether it also predisposes to functional bowel disorders is unclear. Therefore, we assessed the prevalence of IBS-type symptoms in adult celiac patients and correlated this with dietary compliance with gluten exclusion. METHODS: Adult patients (n = 150; 106 women and 44 men) with confirmed celiac disease were randomly selected from a computerized database of >350 patients, and were asked to complete a bowel questionnaire and the Short Form 36 Health Survey (SF-36). The control group (n = 162; 133 women and 29 men) had no history of celiac disease and had similar mean age and sex profile. RESULTS: Of 150 celiac patients reviewed, 30 of 150 (20%) fulfilled the Rome criteria compared with eight of 162 (5%) ontrols. Of the celiac patients 10 of 46 (22%) had partial or no compliance with a gluten-free diet, whereas 20 of 104 patients (19%) adhered to the diet; this difference did not achieve statistical significance. Celiac patients with IBS-type symptoms had a markedly lower quality of life than those without, all eight domains being impaired to a clinically significant extent. Dietary gluten exclusion improved QOL in four of eight scales measured. CONCLUSIONS: The hypothesis that mucosal inflammation in celiac disease may have a sensitizing effect or may predispose to IBS-type symptoms is supported by these results. Failure to attain optimal subjective well-being is common in celiac patients, particularly in those with coexisting IBS. Compliance with a gluten-free diet confers some benefit. PMID- 12094867 TI - Current liver biopsy practices for suspected parenchymal liver diseases in the United States: the evolving role of radiologists. AB - OBJECTIVE: The aim of this study was to determine current liver biopsy practices in the United States. METHODS: We mailed a questionnaire to 260 randomly selected members of the American Association for the Study of Liver Diseases. Statistical analysis was conducted using the chi2 test. RESULTS: A total of 128 members (49%) responded, with 106 responses suitable for inclusion. Of the respondents, 25% had a radiologist perform >50% of their patients' liver biopsies, whereas 38% personally performed all their patients' biopsies. During training, 87% of respondents were taught to perform liver biopsy using the "blind" technique, 2% with ultrasound (US) guidance, and 11% both. A total of 76% used US guidance during liver biopsy. More private practitioners than those practicing in academic centers used the "blind" technique (35% vs 18%; p = 0.02) and referred >50% of their patients for liver biopsy to Radiology (36% vs 17%; p = 0.01); desire for real-time US guidance was the main reason in 43% of cases. Of the respondents, 75% used US for safety reasons; 77% of those who did not use US believed that it was unnecessary. Overall, 87% of respondents indicated that patient safety was the most important issue during liver biopsy; only 2% thought that personal convenience and cost-effectiveness were most important. CONCLUSIONS: Respondents' chief concern was patient safety, and most used US for this reason. Private practitioners were less likely than those practicing in academic centers to use US guidance and were more likely to refer patients to Radiology. PMID- 12094868 TI - Ultrasound-assisted percutaneous liver biopsy performed by a physician assistant. AB - OBJECTIVE: Percutaneous liver biopsy is an essential diagnostic tool utilized in the management of patients with liver disease. This procedure is generally performed by a physician and has a small but well-defined complication rate. We report on the complication rate and efficiency of ultrasound-assisted percutaneous liver biopsy performed by an experienced physician assistant. METHODS: One thousand eighty-six consecutive outpatient liver biopsies (847 hepatic allografts and 239 native livers) were performed at a single center by a physician assistant between June, 1996 and June, 2000. Patients with hepatic mass lesions, unusual hepatic anatomy, and uncorrectable coagulopathy (international normalized ratio > 1.7, platelet count < 50 x 10(9)/L) were excluded. Bedside ultrasonography was used to determine the optimal site for the liver biopsy. Liver biopsies were performed with a 15-gauge Jamshidi aspiration biopsy needle. Patients were observed for 3 h after biopsy, followed by dismissal with subsequent contact in 24 h to assess outcome and complications. RESULTS: Adequate tissue was obtained in 1084 cases (99.8%), with a mean tissue length of 3.2 cm. After the procedure, narcotic analgesia was necessary in 116 (10%) of the patients undergoing liver biopsies. The overall complication rate requiring hospitalization was 0.6%. Major complications requiring intervention occurred in four patients (0.4%). There were no deaths resulting from liver biopsies. CONCLUSION: We conclude that outpatient percutaneous liver biopsy can be safely and effectively performed by a trained physician assistant. PMID- 12094869 TI - Spontaneous bacterial peritonitis caused by Streptococcus bovis: case series and review of the literature. AB - Streptococcus bovis is the rare cause of spontaneous bacterial peritonitis in decompensated cirrhosis. S. bovis bacteremia has long been known to be associated with colon cancer. We describe seven patients and review the seven previous reports of spontaneous bacterial peritonitis patients with S. bovis infection. Most of the patients had cirrhosis and presented with fever, abdominal pain, abdominal distention, and jaundice. Colonic adenomatous polyps with dysplastic change were found in 18.2% of the patients. The approach to this group of patients requires diagnostic paracentesis, blood cultures, ascitic fluid culture, and treatment with antimicrobial agents. Intravenous penicillin is still the antimicrobial agent of first choice (mean minimum inhibitory concentration for penicillin = 0.05 microg/ml). S. bovis is an infrequent cause of spontaneous bacterial peritonitis. The physician could make a case that colonoscopy is not needed because the patient is very sick and the possibility of GI pathology, especially colonic lesions, has been low. However, it may be that colonoscopy should be done if there are clinical suggestions to do so or the patient is well enough to withstand surgery. PMID- 12094870 TI - Does diabetes mellitus influence the perioperative outcome or long term prognosis after resection of hepatocellular carcinoma? AB - OBJECTIVE: This study aims to evaluate whether diabetes mellitus has a significant influence on the perioperative outcome or long term prognosis after resection of hepatocellular carcinoma (HCC). METHODS: The clinicopathological data and postoperative morbidity and mortality of 62 diabetic and 463 nondiabetic patients who underwent resection of HCC between 1989 and 2000 were compared. The long term overall and disease-free survival results were also compared, and the prognostic impact of diabetes mellitus was assessed by multivariate analysis. RESULTS: The diabetic and nondiabetic groups were comparable in terms of the frequency of cirrhosis, liver function, type of resection, and tumor factors such as size and pTNM stage. Overall complication rate (38.7% vs 37.1%, p = 0.820), 30 day mortality (3.2% vs 3.0%, p = 0.583), and hospital mortality (6.4% vs 6.0%, p = 0.782) were similar in diabetic and nondiabetic patients. There was no significant difference in the overall survival (median = 43.5 vs 43.2 months, p = 0.438) or disease-free survival (median = 18.2 vs 15.0 months, p = 0.418). On multivariate analysis, only tumor pTNM stage, operative blood loss, and preoperative indocyanine green retention at 15 min were significant predictors of overall survival. Tumor pTNM stage, size, and operative blood loss were significant predictors of disease-free survival. CONCLUSIONS: This study indicates that diabetes mellitus does not increase the perioperative morbidity or mortality after resection of HCC, nor does it significantly influence the long term prognosis. Based on the current study data, diabetes mellitus should not be considered an unfavorable factor in the selection of patients for resection of HCC. PMID- 12094871 TI - Genetic complexity and serum reactivity of HVR1 quasispecies of hepatitis C virus in patients with cirrhosis. AB - OBJECTIVE: The RNA genome of hepatitis C virus varies considerably, especially within the hypervariable region 1 (HVR1), a domain located on the 5' end of the E2/NS1 envelope region. Our previous study has suggested there were greater numbers of quasispecies in the liver than in matched serum, independent of the viral load. However, the significance of this finding has not been examined extensively at genetic and serological levels. METHODS: By large scale cloning and sequencing, we studied the genetic complexity of HVR1 quasispecies in two selected patients with cirrhosis. The serum reactivity of peptides representing different HVR1 quasispecies isolated from these cases was also estimated by standard ELISA format. RESULTS: We found the same major (dominant and/or subdominant) viral quasispecies variants in serum and in the cirrhotic liver. Genetic analysis suggested that the evolutionary pressure on HVR1 was higher than on its flanking region in quasispecies derived from the liver, whereas this trend is attenuated in quasispecies from serum. The immunoreactivity to peptides representing different HVR1 quasispecies variants showed considerable cross reactivity with heterologous sera, whereas the reactivity was strongest against the dominant HVR1 peptide over time in homologous sera. CONCLUSIONS: These findings indicate that the formation and selection of HVR1 quasispecies may not be driven solely by humoral immune pressure, at least in these two cases. PMID- 12094872 TI - Is NASH underdiagnosed among African Americans? AB - OBJECTIVE: Obesity and type 2 diabetes mellitus are considered risk factors for nonalcoholic steatohepatitis (NASH) and cryptogenic cirrhosis. Because obesity and type 2 diabetes are prevalent among African American females by the 5th and 6th decades, one would expect an increased number of African Americans among patients with NASH and cryptogenic cirrhosis. METHODS: We determined the percentage of patients of African American and European American descent among all of the patients in our liver disease registry and those with NASH and cryptogenic cirrhosis. We also assessed the ethnicity of patients in our registry with other common liver diseases including hepatitis C, and we determined the ethnicity of patients seen at our center with type 2 diabetes and a primary diagnosis of obesity over a 4-yr period. Using census data, we compared these results to our local and regional ethnic demographics. RESULTS: Overall, 199 of 2,253 patients (9%) in the registry were of African American descent, whereas 1,906 were of European American descent (85%). This distribution is similar to the ethnic mix in central Virginia (12% African American, 86% European American) and Albemarle County (12% African American, 83% European American). The prevalence of African American patients among individuals seen at our center for either type 2 diabetes or a primary diagnosis of obesity was over two times the prevalence of African Americans in the county or regional population. In contrast, of 159 NASH patients only one (0.6%) was of African American descent and 154 (97%) were of European American descent (p < 0.001 compared to the total registry, county, or region). Among 206 cryptogenic cirrhosis patients, only two (1%) were of African American descent, whereas 195 (95%) were of European American descent (p < 0.001 compared to the total registry). With regard to other liver diseases, African American patients were slightly overrepresented among hepatitis C patients and markedly overrepresented among patients with hepatic sarcoidosis, similar to previously reported national figures. CONCLUSION: Although there is overrepresentation of African Americans among patients with major risk factors for NASH, individuals of primarily African American descent are infrequently represented among our patients with NASH or cryptogenic cirrhosis. This could result from underrecognition, underreferral, or a true lower prevalence of these disorders among African Americans. PMID- 12094873 TI - Involvement of the Fas system in liver allograft rejection. AB - OBJECTIVE: Recent studies suggest that apoptosis is an important mechanism of cell death in the rejection of liver allografts and that this process is mediated via Fas. The aim of this study was to analyze the expression of the Fas system during the liver allograft rejection and its evolution after treatment. METHODS: We evaluated 14 patients with liver allograft rejection before and after treatment. Fas immunostaining was performed by the labeled streptavidin-biotin peroxidase method using a 200-fold dilution of a monoclonal antibody. Assessment of apoptosis was determined by the terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) technique on deparaffined liver samples. Serum levels of soluble Fas antigen (sFas) were detected by an enzyme immunoassay procedure. Twelve liver transplant patients without allograft rejection were analyzed as a control group. RESULTS: The number of hepatocytes expressing Fas antigen, the percentage of apoptotic hepatocytes, and the sFas levels were higher in patients with liver allograft rejection than in controls (27.9+/-23.1% vs 1.4+/-1.2%, p < 0.001; 2.2+/-0.9% vs 1.0+/-0.1%, p = 0.02; 24.2+/-39.6 vs 2.8+/-4.0 IU/ml, p = 0.03, respectively). There was a correlation between the levels of sFas, AST (r = 0.86, p < 0.001), ALT (r = 0.78, p = 0.02), and gamma-globulin levels (r = 0.86, p < 0.001). After the rejection treatment we found a significant decrease in the Fas antigen expression (18.6+/-13.3%, p < 0.05), TUNEL index (0.2+/-0.4, p < 0.05), and levels of sFas (9.9+/-30.25 IU/ml, p = 0.005). CONCLUSIONS: 1) The demonstration of hepatocytes with Fas antigen expression and the labeling of the nuclei by the TUNEL assay suggest that apoptosis mediated by the Fas system plays a role in the pathogenesis of liver allograft rejection. 2) The Fas expression and the sFas levels decreased in patients with treatment response. PMID- 12094874 TI - Spontaneous Listeria monocytogenes peritonitis: a population-based study of 13 cases collected in Spain. AB - OBJECTIVES: We aimed to assess the incidence, demographic data, clinical features, and outcome of peritoneal infections due to Listeria monocytogenes in individuals with cirrhosis. METHODS: During a 10-yr study period, 153 cases of invasive listeriosis were recorded in a prospective population-based surveillance project carried out in Barcelona, Spain. RESULTS: Thirteen cases were of spontaneous bacterial peritonitis by L. monocytogenes. Ages of the patients ranged between 29 and 85 yr. In addition to cirrhosis, underlying conditions included diabetes mellitus in four and malignancy in three. Bacteremia was present in six cases (46%). Only one patient with bacteremia developed meningitis. Analysis of the peritoneal fluid showed a mean (SD) protein content of 21.5 (9.6) g/L and leukocyte count of 7,273 (9,171) cells/ml. L. monocytogenes serotype 4b was the serogroup predominantly isolated (61%). The mortality rate was 30.7%. Eight patients received empirical antibiotic treatment with cephalosporins. CONCLUSIONS: In geographical areas with a high incidence of listeriosis, L. monocytogenes should be suspected as a causative pathogen of spontaneous bacterial peritonitis in cirrhosis. Early adjustment of antibiotic therapy is essential to reduce mortality. PMID- 12094875 TI - Cirrhosis does not shift the circadian phase of plasma fibrinolysis. AB - OBJECTIVES: The aim of the present investigation was to study the endogenous circadian clock phase rhythm in cirrhotic patients. METHODS: The study population comprised 13 patients with cirrhosis (seven in Child-Pugh class A and six in classes B/C) and nine healthy controls. Plasma melatonin, tissue plasminogen activator antigen, and plasminogen activator inhibitor 1 antigen were measured at 4-h intervals over a 24-h period. Multiple-components rhythmometry using population mean cosinor methods were employed to analyze the findings. RESULTS: All three variables were characterized in both patients and controls by a statistically significant circadian rhythm, with similar profiles. The peak times of tissue plasminogen activator and plasminogen activator inhibitor 1 antigens were practically identical in controls and cirrhotic patients, irrespective of Child-Pugh class (calculated peak at times 6:52, 6:56, and 7:20 for the inhibitor in controls and Child-Pugh class A and classes B/C patients, respectively; p = ns), whereas the peak of melatonin was delayed in classes B/C patients (at times 2:08, 1:56, and 4:00, respectively; p < 0.05). CONCLUSION: The similar circadian phases of plasminogen activator inhibitor antigen in controls and cirrhotic patients in the present investigation indicates that the output rhythm of the internal timekeeping system is not shifted in this pathological condition. PMID- 12094876 TI - Virological profiles in patients with chronic hepatitis C and overt or occult HBV infection. AB - OBJECTIVES: The virological profiles of hepatitis B and C viruses (HBV and HCV) and their interplay in cases of coinfection are undefined. A suppressed and occult HBV infection may occur in hepatitis B surface antigen (HBsAg) negative patients with chronic hepatitis C. The HCV core protein is able to inhibit HBV "in vitro," and serines at positions 99 and 116 are essential for such inhibition. We aimed to assess the HBV and HCV virological profiles in cases of coinfection and to evaluate the relationship between HCV core gene variability and HBV activity. METHODS: Eighty-two anti-HCV positive patients were examined: 35 cases were HBsAg positive, 24 were HBsAg negative with "occult" HBV infection, and 23 were HBV negative. HBV and HCV viremia levels were evaluated in all cases. HCV genomic region coding for the aminoacid sequence 99-116 of core protein was amplified and sequenced in all HCV RNA positive cases. The entire core gene was amplified and sequenced in three randomly selected cases. RESULTS: Serum HCV RNA was detected in all cases but 13, all HBsAg positive individuals; HCV viremia levels of the other 22 HBsAg positive subjects were similar to those detected in HBsAg negative patients with or without occult HBV infection. Among the 35 HBsAg positive patients both HBV DNA and HCV RNA were detected in five cases, HCV RNA alone in 17, and HBV DNA alone in six, whereas seven cases had undetectable levels of both viruses. Sequencing analyses showed that the HCV core gene was highly preserved in all patients. CONCLUSION: A wide spectrum of HCV and HBV virological patterns may occur in a case of coinfection. HCV core variability is not related to HBV activity "in vivo." PMID- 12094877 TI - New occurrence and recurrence of neoplasms within 5 years of a screening colonoscopy. AB - OBJECTIVE: The fear that colorectal adenomas were missed on initial colonoscopy or that new adenomas have developed is often a rationale for repeating a colonoscopic examination. The aim of this study was to delineate risk factors associated with recurrence of colorectal adenomas after an initial baseline screening colonoscopy. METHODS: The study population comprised 875 subjects who underwent a baseline screening colonoscopy followed by a second examination 1-5 yr later. Multiple logistic regression was used to assess the influence of potential risk factors on the occurrence or recurrence of colorectal adenomas, the strength of the influence being expressed as an OR with a 95% CI. RESULTS: Colorectal adenomas were detected in 484 of all patients (55%) at baseline colonoscopy. Within a 1- to 5-yr time interval, 181 patients (37%) had recurrent adenomas (adenomas were removed during the first colonoscopy) and 73 patients (19%) had newly developed adenomas (adenomas were absent on the first colonoscopy). The occurrence of adenomas at baseline screening colonoscopy was the only factor associated with an increased risk for the recurrence of adenomas at follow-up (OR = 2.51, 95% CI = 1.77-3.55). Recurrence was associated with multiple baseline adenomas (4.45, 2.98-6.64) and baseline adenomas larger than 1 cm (2.62, 1.99-3.11). Recurrence was not associated with histology type or family history of colorectal cancer. There was a significant trend for adenomas to recur in the same proximal or distal segment as the baseline adenomas (p = 0.02). CONCLUSIONS: Colon adenomas tend to recur with greater frequency if the adenomas removed at baseline were either large or multiple. Although patients with large adenomas or multiple adenomas at baseline screening colonoscopy are at a 2.6- to 4.5-fold risk for recurrence of adenomas, the rate of de novo adenoma formation in patients without baseline adenomas may be large enough to warrant repeat colonoscopy at some time in the future. The exact timing of the follow-up colonoscopy needs to be determined. PMID- 12094878 TI - Step section histology of proximal colon polyps that appear hyperplastic by endoscopy. AB - OBJECTIVE: Recent studies have identified a group of polyps, usually located in the proximal colon, with distinct endoscopic features that predict hyperplastic histology. These polyps may occasionally have foci of dysplastic (adenomatous) tissue. We aimed to define the prevalence of adenomatous foci in these polyps. METHODS: Fifty-one consecutive hyperplastic-appearing proximal colon polyps were sectioned at 1-mm intervals for histological analysis. RESULTS: All 51 polyps were hyperplastic, but two (4%) had foci of adenomatous tissue. CONCLUSION: The prevalence of adenomatous foci in proximal colon polyps with endoscopic features of hyperplastic polyps is low. Whether endoscopic resection of polyps with these features is warranted remains uncertain. PMID- 12094879 TI - Impact of physician specialty on the cost of nonvariceal upper GI bleeding care. AB - OBJECTIVE: Upper GI bleeding (UGIB) is a common medical emergency that leads to a high consumption of medical resources and costs. We aimed to analyze the influence of physician specialty on the costs of nonvariceal UGIB care. METHODS: We retrospectively assessed 350 nonvariceal UGIB episodes that were primarily cared for by gastroenterologists (n = 142), internists (n = 67), or surgeons (n = 141). Gastroenterologists followed evidence-based clinical protocols that included early endoscopy and early hospital discharge for uncomplicated bleeding. A risk score system was used to control for severity of illness. Linear regression analyses were performed to find out predictors of costs and the influence of specialist care on length of stay (LOS). RESULTS: The overall mean hospital cost was significantly lower in patients cared for by gastroenterologists (EUR 1,630) than in those managed by internists (EUR 3,745, p < 0.001) or surgeons (EUR 2,513, p < 0.05). The mean LOS was the variable with highest influence on total cost. Patients cared for by gastroenterologists had a mean LOS significantly shorter (7.3 days) than that of those treated by internists (16.2 days, p < 0.001) or surgeons (11 days, p < 0.001). Hospital costs and LOS differences were maintained when adjusting for severity of illness. In caring for low risk patients, nongastroenterologists had a higher probability of having a hospital stay longer than 4 days (odds ratio = 18.4, Cl = 4.6-73.6, p < 0.001). CONCLUSION: The implementation of specific evidence-based protocols by gastroenterologists reduces length of hospital stay and saves medical costs in patients with nonvariceal UGIB, especially those at low risk. PMID- 12094880 TI - Medicine spoon. PMID- 12094881 TI - Severe hemolysis and renal failure in glucose-6-phosphate dehydrogenase deficient patients with hepatitis E. AB - Hemolytic anemia as a complication of acute hepatitis is not uncommon in patients with glucose-6-phosphate dehydrogenase deficiency. However, severe hemolysis in these patients is rare. We report a cohort of five patients with acute viral hepatitis E who developed severe intravascular hemolysis and unusually high levels of bilirubin. All five patients had severe, complicated, protracted courses of illness. Four patients developed acute renal failure, and two of these required hemodialysis. To the best of our knowledge this is the first report of a cohort of patients with glucose-6-phosphate dehydrogenase deficiency and acute viral hepatitis E with severe intravascular hemolysis. We emphasize the fact that intravascular hemolysis should be suspected in patients with acute viral hepatitis E with marked bilirubinemia and anemia. Measures to prevent renal failure should be taken in such cases. PMID- 12094882 TI - Botulinum toxin for the treatment of gastroparesis: a preliminary report. AB - Gastroparesis is a disorder of gastric motility that results in delayed gastric emptying. Common symptoms include early satiety, postprandial fullness, epigastric pain, nausea, vomiting, and weight loss. The underlying etiologies of gastroparesis are many and include diabetes, prior gastric surgery, collagen vascular disorders, and a previous viral illness. Up to one third of cases are classified as idiopathic. Treatment typically consists of a change in diet to small volume, frequent meals and the use of the prokinetic agents metoclopramide, cisapride, erythromycin, or domperidone. Botulinum toxin has recently been shown to be effective in treating disorders of smooth muscle hypertonicity in the GI tract. This case report describes three patients with severe gastroparesis whose symptoms persisted despite dietary changes and the use of high dose prokinetic agents. All three were treated with intrasphincteric injection of the pylorus with botulinum toxin and all had significant symptomatic improvement afterwards. Possible mechanisms of action of botulinum toxin on the pylorus and its effects in patients with gastroparesis are discussed. PMID- 12094883 TI - Hydatid disease in pregnancy. PMID- 12094884 TI - Nutcracker esophagus: GERD or an esophageal motility disorder. PMID- 12094885 TI - Antibiotic prophylaxis in acute necrotizing pancreatitis. PMID- 12094886 TI - The approach to high grade dysplasia in Barrett's esophagus. PMID- 12094887 TI - Implication of detecting mucosal dysplasia in a patient with ulcerative colitis. PMID- 12094888 TI - Economic impact of hospitalization for end-stage liver disease. PMID- 12094889 TI - Chronological change in H. pylori antibody titers after treatment and its utility in follow-up. PMID- 12094890 TI - Stump appendicitis diagnosed by colonoscopy. PMID- 12094891 TI - Screening for Barrett's esophagus: searching for a new technique. PMID- 12094892 TI - Successful management of acute portal vein thrombosis by low molecular weight heparin and oral anticoagulation. PMID- 12094893 TI - Autoimmune hepatitis preceded by acute hepatitis A infection. PMID- 12094894 TI - Precipitation of dysphagia due to achalasia cardia by hemorrhagic stroke. PMID- 12094895 TI - HFE gene mutations prevent sustained virological response to interferon plus ribavirin in chronic hepatitis C patients with serum markers of iron overload. PMID- 12094896 TI - Gangrenous cholecystitis after colonoscopy. PMID- 12094897 TI - Endoscopic diagnosis of malignant melanoma metastatic to the stomach. PMID- 12094898 TI - Is low male sex ratio in offspring of celiacs an advantage? PMID- 12094899 TI - Nitric oxide production in rectal dialysate is a marker of disease activity and location in children with inflammatory bowel disease. PMID- 12094900 TI - Helicobacter pylori and SIDS: the jury is in at last! PMID- 12094901 TI - Early phase mantle cell lymphoma: macroscopic similarities to terminal ileal Crohn's disease. PMID- 12094902 TI - Pantoprazole-induced recurrent anaphylactic shock. PMID- 12094903 TI - A preliminary study on the reliability of physical performance measures in older day-care center clients with dementia. AB - BACKGROUND: Decline in physical functional ability is an intrinsic component of the dementia syndrome. Reductions in muscle mass and strength represent a major factor in the loss of functional ability Although resistance exercise has been studied as a method for maintaining/recovering function in populations of frail older adults, people with dementia have been systematically excluded because of uncertainty about the reliability of outcome measurements. OBJECTIVE: The purpose of this study was to determine the test-retest reliability of a battery of established performance-based measures of strength and function among subjects with dementia. SETTING: A hospital-affiliated adult day-care facility. SUBJECTS: Twelve of 28 older subjects with dementia of various etiologies were available for two assessments prior to implementation of a resistance-exercise intervention. METHODS: Subjects underwent an assessment of lower extremity strength and physical function consisting of two recorded trials of bilateral isometric strength of the knee extensor, hip flexor, and dorsiflexor muscles, as well as hand grip strength; repeated chair stands, evaluation of usual- and maximal safe-gait speed over a 6-m course, and the Timed-Up-and-Go Test. The entire assessment was repeated approximately 1 week later. An average of the trials for each measurement was computed for each of the two assessment periods, and intraclass correlation coefficients (ICCs) for these paired measurements were estimated using STATA. RESULTS: ICCs ranged from .56 for left iliopsoas to .77 for left dorsiflexors among the strength measures whereas measures of function ranged from .80 for number of steps in usual gait to .95 for time of fast gait. CONCLUSIONS: Performance-based measures of strength and function can be reliably assessed in older people with dementia, although measures of function appear to be more reliable than measures of strength. PMID- 12094904 TI - Impact of training dementia caregivers in sensitivity to nonverbal emotion signals. AB - Ninety-one mid- to late-stage dementia patients residing in nursing homes, along with their staff caregivers, participated in a study designed to assess whether training caregivers in sensitivity to nonverbal communication could enhance mood and reduce symptoms in patients and improve psychological well-being in caregivers. Patients and staff at three nursing homes comprised three groups that were randomly assigned to either a nonverbal sensitivity group, a behavioral placebo group that received instruction in the cognitive and behavioral aspects of dementia, and a wait-list control. Training consisted of 10 one-hour sessions taught by a clinical psychologist using prepared materials. Patient measures, which were taken at baseline and at 4 three-week intervals, included patient symptomatology (depression, agitation, behavioral symptoms), as reported by the staff caregivers, and positive and negative facial expressions of emotion elicited during a face-to-face interview and coded by trained research staff. Results indicated that positive affect increased sharply during the first 6 weeks after intervention in the nonverbal group, with the placebo and wait-list controls showing no change. There was also a decline in negative affect across time for all groups. Effects with respect to patient symptomatology did not reach significance. Caregivers in both training groups showed a decline in symptomatology, whereas the wait-list control group did not. PMID- 12094905 TI - Public health models of mental health care for elderly populations. PMID- 12094906 TI - Carer impressions of participation in Alzheimer's disease clinical trials: what are their hopes? And is it worth it? AB - BACKGROUND: We aimed to assess why carers seek participation for their relatives in clinical trials of Alzheimer's disease (AD) medications, and to assess carer impressions of the value of trial participation. METHOD: A retrospective questionnaire was sent to 31 carers of participants in clinical trials for AD conducted at the Mental Health Research Institute and the National Aging Research Institute between January 1997 and December 1999. RESULTS: Twenty-five questionnaires (81%) were returned, completed to an extent to permit analysis. Helping a relative feel better, contributing to medical science, and hoping for a cure were the main reasons for trial participation. In general, carers found the regular consultation with research staff supportive and would recommend trial participation to others. CONCLUSION: A greater understanding of the motivations of carers may aid the recruitment and retention of participants in clinical trials. PMID- 12094907 TI - Spouse caregivers' perceptions of influence of dementia on marriage. AB - OBJECTIVES: To investigate what kind of changes spouse caregivers of demented patients experience after the onset of dementia (a) in the general atmosphere, happiness, and relations of marriage and (b) in the sexual side of marriage. DESIGN: Semistructured telephone interviews of spouse caregivers of demented patients. SETTING: Community-living demented patients and their spouse caregivers in eastern Finland. PARTICIPANTS: The spouse caregivers of 42 demented patients recruited from a previous intervention study. MEASURES: The questionnaire covered different areas of marriage from the time before and after the onset of dementia. RESULTS: A statistically significant decline had occurred in extent of happiness (p = .012), in equal relations (p = .001), and in patients' expressions of sexual needs (p < .001) when compared the time before and after dementia. Twenty-five (60%) of the caregivers reported that the demented patient had shown at least one negative sexual behavioral change during the course of dementia. Seven male patients (24%) had shown the behavioral symptom of constantly expressing need for making love. One in 10 caregivers had experienced positive sexual behavioral changes. In one third of the patients, the expressions of tenderness towards the caregiver had increased. Dementia did not affect significantly the general atmosphere of the marriage. Out of those still in home care, at 3 years from the onset of dementia, 19 couples (46%) continued to practice intercourse, at 5 years the number was 15 couples (41%), and at 7 years it had declined to 7 couples (28%). CONCLUSIONS: Dementing illness has a major negative impact on many dimensions of marriage. However, there are also positive changes and preserved aspects of marriage. Dementia seems to have a surprisingly little impact on whether the couple continues to have intercourse when compared with the general aging population. PMID- 12094908 TI - Volumetric MRI measurements can differentiate Alzheimer's disease, mild cognitive impairment, and normal aging. AB - BACKGROUND: Volumetric magnetic resonance imaging (MRI) has been extensively studied in the last decade as a method to help with the clinical diagnosis of Alzheimer's disease (AD). In recent years, researchers have also started investigating if that technique would be useful to identify individuals with mild cognitive impairment (MCI), differentiating them from AD patients and from normal elderly controls. This research project was planned to assess the accuracy of volumetric MRI to differentiate those groups of individuals. METHOD: The investigation involved 39 patients with diagnosis of mild to moderate dementia in AD, according to the criteria of the NINCDS-ADRDA, DSM-III-R, and ICD-10; 21 subjects with complaints of cognitive decline without other psychiatric disorders (MCI); and 20 normal elderly controls. All the subjects were submitted to a standard protocol, including volumetric MRI evaluations. RESULTS: The results indicated that all regions of interest measured (amygdala, hippocampus, and parahippocampal gyrus) were significantly different (p < .005) in AD patients compared to MCI subjects and controls. The left volumetric measures (amygdala, hippocampus, and parahippocampal gyrus) were also significantly different between the MCI subjects and controls (p < .05). The discriminant function analysis correctly classified 88.14% of the AD patients and controls, 81.67% of AD patients and MCI subjects, and 80.49% of the MCI subjects and controls. CONCLUSIONS: The results suggest that measures of medial temporal lobe regions are useful to identify mild to moderate AD patients and MCI subjects, separating them from normal elderly individuals. PMID- 12094909 TI - Endogenous antioxidant activities in relation to concurrent vitamins A, C, and E intake in dementia. AB - Previous reports on the activities of essential endogenous antioxidants such as superoxide dismutase, catalase, and glutathione in dementia patients have not included a simultaneous quantitative assessment of dietary antioxidant intake. This is important because the reported differences in endogenous antioxidant levels among dementia patients may have reflected variations in the total antioxidants' intake. In this study we measured the levels of antioxidant vitamins A, C, and E in the diet of 81 dementia patients and controls at the same time as assessing blood levels of three endogenous antioxidants. Results showed a significant decrease in the intake of vitamins C (p < .001) and E (p < .01) in patients with severe Alzheimer's disease (AD) when compared to controls. Patients with mild/moderate AD differed from controls only in the intake of vitamin C (p < .01). The blood levels of catalase but not superoxide dismutase and glutathione were significantly decreased in the patients with severe AD when compared to controls (p < .01), patients with mild/moderate AD (p < .0 1), and patients with dementia with Lewy bodies (p < .05). The blood catalase levels of dementia patients, as a whole, were significantly and positively associated with the intake of vitamins A (p < .05), C (p < .01), and E (p < .05). The results indicated that dietary intake of vitamins A, C, and E may influence blood levels of catalase possibly through their antioxidant effects on free radicals. The data underscore the importance of concurrent quantitative assessment of nutritional intake when measuring endogenous antioxidant activities and support a role for antioxidant supplementation in the treatment of dementia disorders. PMID- 12094910 TI - Sleep/wake cycle disturbance in Alzheimer's disease: how much is due to an inherent trait? AB - Major advances in understanding the physiology and genetics of circadian rhythm in the past decade challenge the researcher of sleep/wake disorders in Alzheimer's disease (AD) to distinguish patient characteristics stable across the course of illness ("traits") from characteristics that vary with stage of illness ("states"). A components-of-variance approach with a repeated measures model was used to examine the between-subjects variance over time ("trait") vs. within subjects ("state") variance in 42 patients with probable AD followed, on average, over 2 years on actigraphic sleep/wake measures. Mental status scores indexed stage of illness. Actigraphic measures of sleep efficiency and circadian rhythmicity appeared predominantly "trait," with between-individual differences accounting for over 55% of variance compared to the less than 5% of variance related to stage of cognitive impairment. We discuss how "state-trait" analyses can be helpful in identifying areas of assessment most likely to be fruitful objectives of physiologic and genetic research on sleep/wake disturbance in AD. PMID- 12094911 TI - Everyday problem solving in African Americans and European Americans with Alzheimer's disease: an exploratory study. AB - In this exploratory study, we compared the performance of 10 African American and 26 European American persons with early- to mid-stage Alzheimer's disease (AD) to 20 nondemented elderly (NE), using a shortened version of the Test of Problem Solving (TOPS). The TOPS measures verbal reasoning to solve everyday problems in five areas: explaining inferences, determining causes, answering negative why questions, determining solutions, and avoiding problems. Six linguistic measures were also examined: total utterances, abandoned utterances, length of utterances, maze words, questions, and total words. NE performed better than AD subjects on all but one measure of verbal reasoning ability. AD subjects also showed a trend to use more total utterances and abandoned utterances than NE. For the AD group, no ethnic differences were found for verbal reasoning or linguistic measures. The findings from this preliminary investigation suggest that, compared to European Americans, African American persons with AD demonstrate similar everyday problem solving and linguistic skills. Thus, assessments such as TOPS that examine everyday problem solving may be a useful nonbiased evaluation tool for persons with AD in these two ethnic groups. PMID- 12094912 TI - Elderly patients with deliberate self-poisoning treated in an Australian general hospital. AB - OBJECTIVE: To examine the demographic, prescription, ingestion, and psychiatric diagnostic factors that distinguished elderly from nonelderly patients treated for deliberate self-poisoning (DSP). METHOD: A prospective case series study of 2,667 patients presenting to a regional referral center for poisoning (Newcastle Mater Hospital, NSW, Australia), January 1991 to July 1998. The sample was stratified into two groups, 65 years or greater (n = 110) and 64 years or less (n = 2,557) at the time of index admission. The groups were compared using a forward stepwise logistic regression model. Uncontrolled comparisons were analyzed by chi square statistic with Bonferroni-adjusted p values and controlled comparisons by odds ratio (OR) with 95% confidence interval (CI). RESULTS: The elderly group represented 4.1% of the total. The logistic regression analysis found the elderly DSP group was more likely to have a longer length of stay (OR 5.90, CI 3.87 9.00), to have been prescribed "other" drugs (neither benzodiazepines, mood treatment drugs, nor paracetamol) before admission (OR 5.32, CI 3.34-8.48), to have been prescribed benzodiazepines (OR 3.15, CI 2.03-4.89), and to be diagnosed with major depression (OR 2.17, CI 1.41-3.36) than the younger group. The elderly group was less likely to have ingested paracetamol (OR 0.28, CI 0.14-0.54) or "other" drugs (neither benzodiazepines nor mood treatment drugs) in the DSP episode (OR 0.33, CI 0.20-0.54). DISCUSSION: Elderly DSP patients differ in several important respects from younger patients. They have higher morbidity as a result of the DSP. Major depression plays a more important role. The strong relationship between benzodiazepine prescription and DSP in the elderly raises questions and possible prevention strategies. PMID- 12094913 TI - Acid-base management--is it relevant for the study design of hypothermic neuroprotection? PMID- 12094914 TI - Pathophysiology of cocaine abuse. AB - Cocaine abuse is increasing, particularly among young people. It is no longer a drug used exclusively by higher socio-economic groups. This review highlights the dangers of both acute and chronic cocaine abuse and its lethal potential even on the first encounter. The systemic effects of cocaine are described along with the possible pathology that may present itself to the attending clinician. PMID- 12094915 TI - Operating room management and strategies in Switzerland: results of a survey. AB - BACKGROUND AND OBJECTIVE: Operating room management structures and interrelationships both within the operating suite and with other departments in the hospital can be very complex. Several different professional and support groups are represented that often have infrastructures of their own that may compete or conflict with the management hierarchy in the operating room. Today, there is often little actual management of the operating suite as an entity. We surveyed current operating room management in Switzerland. METHODS: A questionnaire was sent to the chief anaesthesiologists of all public hospitals in Switzerland. It asked for information about the structure, organization and management of operating rooms as well as respondents' opinions and expectations about management. Derived from both the literature and the results of the survey, a 'stages of excellence' model of best practice was developed. RESULTS: The overall response rate was 70%. Most anaesthesiologists were unsatisfied with current management policies and structures in their operating rooms. Of the hospitals questioned, 40% did not have an information system at all for the operating rooms. The remaining 60% had an information system that allowed rough scheduling in 71%, but only a few had more sophisticated systems that enabled dynamic scheduling (19%), user-defined conflict checking (5%), administration of a subsequent patient transfer station (postanaesthesia care units, intensive medical care, intensive care units) (10%) or other more advanced uses. All hospitals questioned offered some type of ambulatory surgery in a 'hospital integrated' manner (i.e. use of the same operating room for both in- and outpatient surgery), but none had implemented a more efficient system where outpatient surgery was performed in separate facilities. CONCLUSIONS: Current management of the operating room in Switzerland is far from best-practice standards. PMID- 12094916 TI - Augmented spontaneous breathing and pulmonary gas exchange during pneumoperitoneum. AB - BACKGROUND AND OBJECTIVE: Ventilation of the lungs with positive end-expiratory pressure during pneumoperitoneum has been shown to improve the arterial partial pressure of oxygen. The implications of spontaneous breathing on pulmonary gas exchange remain unknown in this setting. We therefore sought to examine the influence of pressure-support ventilation with spontaneous breathing on gas exchange during simulated laparoscopy. METHODS: Ten pigs were subjected to pneumoperitoneum at a pressure of 15 cmH2O. Animals received, in a random order, pressure-support and pressure-controlled ventilation for 60 min per mode. Inert gas and haemodynamic measurements were performed before changing to a subsequent mode. RESULTS: Pressure-support ventilation was more efficient than pressure controlled ventilation regarding perfusion of normal V(A)/Q lung areas (78 +/- 4% vs. 72 +/- 5%) (P < 0.05), alveolar-arterial partial pressure of oxygen difference (9.73 +/- 1.3 vs. 11.2 +/- 1.2 kPa) and arterial partial pressure of oxygen (14.93 +/- 1.6 vs. 13.7 +/- 2.0 kPa) (P < 0.05). CONCLUSIONS: Pressure support ventilation resulted in significantly better gas exchange than pressure controlled ventilation in this model of simulated laparoscopy. PMID- 12094917 TI - Effects of induction of anaesthesia with sufentanil and positive-pressure ventilation on the intra- to extrathoracic volume distribution. AB - BACKGROUND AND OBJECTIVE: Induction of general anaesthesia in combination with positive-pressure ventilation is often associated with a significant decrease of arterial pressure. A decreased preload may contribute to this phenomenon. The aim was to investigate whether a change in cardiac filling occurs following the induction of general anaesthesia with sufentanil under typical clinical conditions. METHODS: Fifteen patients scheduled for elective coronary bypass grafting were studied immediately before surgery. In addition to standard monitors, a transpulmonary double-indicator dilution technique measured in vivo intrathoracic blood volume, global end-diastolic volume and total circulating blood volume. For induction of anaesthesia 2 microg kg(-1) sufentanil was given. Measurements were performed awake and after the induction of anaesthesia, intubation and mechanical ventilation of the lungs. RESULTS: To maintain arterial pressure during the induction period within -20% of baseline pressure, on average 22 +/- 6mLkg(-1) crystalloids and 8 +/- 6mLkg(-1) colloids were given. Despite these amounts of fluid, cardiac filling was decreased, whereas circulating blood volume increased significantly. Both central venous pressure and pulmonary capillary wedge pressure increased. CONCLUSIONS: Induction of general anaesthesia with positive-pressure ventilation is regularly associated with a blood volume shift from intra- to extrathoracic compartments. Even in low-dose opioid monoanaesthesia with sufentanil--often regarded as relatively inert in haemodynamic terms--the phenomenon could be demonstrated as the primary cause of the often-observed decrease of arterial pressure. It seems, therefore, rationally justified to restore cardiac filling by generous administration of intravenous fluids, at least in patients with unaffected cardiac pump function. During induction of anaesthesia, central venous pressure and pulmonary capillary wedge pressure do not reliably indicate cardiac filling. PMID- 12094918 TI - Psychomotor performance after short-term anaesthesia. AB - BACKGROUND AND OBJECTIVE: The aim was to examine the immediate effects of short term anaesthesia on the different components of psychomotor performance of the upper extremity and cognitive functions, and to find out if there were any differences in the sensitivities of the different tests. The measured psychomotor aspects were simple reaction time, choice reaction time, speed of movement, index finger-tapping speed, co-ordination, visual spatial memory capacity, digit-symbol substitution and the Maddox Wing test. METHODS: The subjects were 30 female patients aged 24-50 yr who had been through a minor gynaecological operation. Anaesthesia had been induced with propofol and alfentanil. The measurements were mainly made with the HPM/BEP system, and the tests were performed 1 h before the anaesthesia and immediately after the wake-up. RESULTS: Short-term anaesthesia prolonged the simple reaction time by 7% and the choice reaction times by 25% (one-choice) and 7% (two-choice) and decreased the speed of movement by 10% (one choice) and 19% (two-choice), index finger-tapping speed by 7% and co-ordination by 7%. In addition, visual spatial memory capacity decreased by 21%, digit-symbol substitution increased by 5% and the Maddox Wing test increased by 68%. CONCLUSIONS: Based on the results, it seems that short-term anaesthesia reduces both signal processing at the central level, and motor control and co-ordination of movements at the peripheral level, and has a decreasing effect on motor performance in the above-mentioned aspects measured immediately after wake-up. PMID- 12094919 TI - Haemodynamic and splanchnic organ blood flow responses during sevoflurane-induced hypotension in dogs. AB - BACKGROUND AND OBJECTIVE: The safety of hypotension induced by sevoflurane and splanchnic organ blood flow remains to be clarified. The aim was to investigate the effects of sevoflurane-induced hypotension on systemic haemodynamics and splanchnic organ blood flows in dogs. METHODS: Mean arterial pressure was maintained at 60 mmHg by increasing sevoflurane concentrations. The renal, hepatic and pancreatic blood flows were measured by using the hydrogen clearance method. RESULTS: Hypotension induced by sevoflurane resulted in a 50% decrease of mean arterial pressure due to a 30% reduction in systemic vascular resistance associated with a 30% decrease in cardiac index. The mechanisms causing the lower cardiac index were produced by the decreases in heart rate and left ventricular dP/dt(max). Renal, hepatic and pancreatic blood flow were reduced, but the whole body oxygen consumption did not change during the hypotensive period. CONCLUSIONS: The haemodynamic changes induced by sevoflurane were caused by the suppression of arterial baroreflexes and myocardial depression, but splanchnic organ blood flows, though reduced, could provide adequate peripheral perfusion to meet the decrease in oxygen supply. PMID- 12094920 TI - Predictors of successful neuraxial block: a prospective study. AB - BACKGROUND AND OBJECTIVE: The epidural and subarachnoid spaces should be identified at the first attempt, since multiple punctures increase the risk of postdural puncture headache, epidural haematoma and neural trauma. The study aimed to describe the predictors of successful neuraxial blocks at the first attempt. METHODS: After institutional Review Board approval, 1481 patients undergoing spinal or epidural anaesthesia were prospectively enrolled. For each block we recorded: gender, age, height, weight, body habitus, anatomical landmarks (palpability of the spinous processes), spinal anatomy, patient positioning, premedication, needle type and gauge, approach, spinal level of the block, and the provider's level of experience. Retrieval of cerebrospinal fluid or loss of resistance to saline or air identified the subarachnoid and epidural spaces, respectively. The outcome variable was the first attempt success or failure (whether or not the needle was correctly located with one skin puncture and produced adequate surgical anaesthesia). Backward stepwise logistic regression tested its association with the other variables. RESULTS: The first attempt rate of success was 61.51%. Independent predictors (Odds ratio, 95% confidence limits) were the quality of anatomical landmarks (1.92 (1.57; 2.35)), the provider's level of experience (1.24 (1.15; 1.33)) and the adequacy of patient positioning (3.84 (2.84; 5.19)). CONCLUSIONS: The successful location of the subarachnoid or the epidural space at the first attempt is influenced by the quality of patients' anatomical landmarks, the adequacy of patient positioning and the provider's level of experience. PMID- 12094922 TI - Bacterial contamination of propofol: the effects of temperature and lidocaine. AB - BACKGROUND AND OBJECTIVE: The intravenous anaesthetic propofol may become contaminated once the ampoules have been opened. The effect of lidocaine and cooling was tested on the bacterial contamination of propofol. METHODS: The study was performed in two parts. In Part 1,1920 aliquots of propofol alone, and of a propofollidocaine mixture, were drawn into sterile syringes and stored at room temperature (24-26 degrees C) or in the refrigerator (12-14 degrees C). In Part 2, 1200 aliquots from opened ampoules of propofol alone, or as a propofol lidocaine mixture, were stored at room temperature or in the refrigerator. Samples were aerobically cultured at 0, 1, 2, 4, 8 and 12 h. RESULTS: In Part 1, diphtheroid bacillus was isolated from one aliquot (0.06%). In Part 2, there was bacterial growth in both groups; the number of contaminated ampoules increased with time and it was 20-26% at 12 h. Diphtheroid bacilli and coagulase-negative staphylococci were the most frequent micro-organisms. CONCLUSIONS: When propofol is stored in opened ampoules, the bacterial contamination rate is high. Adding lidocaine, or storing opened ampoules at 12-14 degrees C, does not affect the contamination rate, except during the first few hours. It is advisable to draw propofol aseptically into a syringe in an amount that can be used during one procedure. PMID- 12094921 TI - Effect of aminophylline on recovery from sevoflurane anaesthesia. AB - BACKGROUND AND OBJECTIVE: In this randomized, double-blind study, we aimed to investigate the effect of aminophylline on recovery from sevoflurane. METHODS: One-hundred ASA I-II patients scheduled for elective surgery were randomly divided into two groups receiving either NaCl 0.9% (Group 1, n = 50) or aminophylline 5mg kg(-1) (Group 2, n = 50). All patients were premedicated with atropine 0.01 mgkg(-1) and midazolam 0.06mgkg(-1) intramuscularly. Anaesthesia was induced with propofol 2mg kg(-1) for muscle relaxation, and atracurium 0.5 mg kg(-1) was maintained with sevoflurane 2% in 50% oxygen and nitrous oxide. Further atracurium (0.1 mgkg(-1) was given when needed. Aminophylline or saline was given after sevoflurane was discontinued. Heart rate, mean arterial pressure, peripheral oxygen saturation, the duration of anaesthesia and recovery times (eye opening, verbal response, extubation and successful performance of arithmetical calculations) were recorded. RESULTS: There were no statistically significant differences in mean arterial pressure, peripheral oxygen saturation and anaesthesia time between the two groups. Heart rate increased significantly (P < 0.05) after aminophylline and was also higher than in the placebo group. Recovery times were significantly shorter (P < 0.001) in the patients receiving aminophylline. CONCLUSIONS: Aminophylline speeded recovery after sevoflurane anaesthesia and it may have some advantage in anaesthesia practice for patients. PMID- 12094923 TI - Differentiating massive carbon dioxide embolism from tension pneumothorax during laparoscopic cholecystectomy. PMID- 12094924 TI - Ventricular asystole and complete heart block after thoracic epidural analgesia for thymectomy. PMID- 12094925 TI - Rapid, ultra-sensitive detection of gas phase elemental mercury under atmospheric conditions using sequential two-photon laser induced fluorescence. AB - We have examined the sensitivity of sequential two photon laser induced fluorescence (LIF) detection of elemental mercury, Hg(0) in the gas phase. The most sensitive approach involves an initial laser excitation of the 6(3)P1-6(1)S0 transition at 253.7 nm, followed by excitation with a second laser to the 7(1)S0 level. Blue shifted fluorescence is observed on the 6(1)P1-6(1)S0 transition at 184.9 nm. The excitation scheme, involving sequential excitation of two atomic transitions, followed by detection of the emission from a third is extremely specific and precludes detection of anything other than atomic mercury. Using our 10 Hz laser system we have achieved a detection sensitivity of 0.1 ng m(-3) at a sampling rate of 0.1 Hz, i.e. averaging 100 laser shots at a pressure of one atmosphere in air. At low concentrations we sampled simultaneously with an automated mercury analyzer (Tekran 2537A), to ensure accuracy. We have examined the linearity of the technique, generating flows containing mercury concentrations between 1 and 10,000 ng m(-3) using a permeation tube and dynamic dilution, but relying on the concentrations given by the Tekran at low levels and the concentration calculated from dilution at high levels. We find that the detection is linear over the five orders of magnitude that we were able to vary the concentration. Our measured detection limits in He and Ar are much lower as these gases are inefficient fluorescence quenchers. PMID- 12094926 TI - The 21st century analyst: developments in data analysis and visualisation. PMID- 12094927 TI - The heterogeneous composition of working place aerosols in a nickel refinery: a transmission and scanning electron microscope study. AB - Size, morphology and chemical composition of individual aerosol particles collected in a nickel refinery were analyzed by scanning electron microscopy and energy-dispersive X-ray microanalysis (EDX). The phase composition was determined by selected area electron diffraction and EDX in a transmission electron microscope. Most particles are heterogeneous on a nanometer scale and consist of various phases. Nickel phases observed in the roasting and anode casting departments include metallic nickel, bunsenite (NiO), trevorite (Ni,Cu)Fe2O4, heazlewoodite Ni3S2, godlevskite (Ni,Cu)9S8, orthorhombic NiSO4 and an amorphous Ni,Cu.Al,Pb sulfate of variable composition. Additional phases encountered include corundum (Al2O3), murdochite (PbCu6O8), hexagonal Na2SO4, anhydrite (CaSO4), graphite (C) and amorphous carbon. The implications of the occurrence of the different Ni phases and their nanometer size for the study of adverse health effects are explored. PMID- 12094928 TI - Evaluation of the NIOSH draft method 5525 for determination of the total reactive isocyanate group (TRIG) for aliphatic isocyanates in autobody repair shops. National Institute for Occupational Safety and Health. AB - This paper evaluates the performance of the NIOSH draft method 5525 for analysis of monomeric and TRIG aliphatic isocyanates in autobody repair shops. It was found that an optimized pH gradient enhanced noticeably the resolution and, therefore, identification of aliphatic isocyanates. Samples proved to be very stable for at least a year when stored at -13 degrees C in the freezer, and no major stability problems were found for the MAP reagent. The detector response factor RSD for selected MAP ureas was 40% in the fluorescence (FLD), 3% in the UV at 254 nm (UV254), and 1% in the UV at 370 nm (UV370). The mean FLD/UV254 and UV254/UV370 detector response ratios of standards were 31.7 (RSD = 37.8) and 17.1 (RSD = 5.4), respectively. The FLD/UV254 ratio in bulks varied from 0.41 to 1.97 times the HDI monomer ratio. The mean UV254/UV370 ratio in bulks was 16.1 (range 14.1 to 19.2, N = 38). Mean (range) recovery of 92 (91.2-93.2)% was found for the N3300 (isocyanurate) spiked on 25 mm quartz fiber filters in the range 0.07 to 2.2 microg NCO ml(-1). Mean (range) recovery for impingers was 100.7 (91.7 106.0)% for N3300 in the concentration range of 0.018 to 2.5 microg NCO ml(-1) and 81.0 (76.1-89.1)% for IPDI in the concentration range of 0.016 to 1.87 microg NCO ml(-1). Analytical method precision was 3.4% and mean bias 7.4% (range = 0 25%). The NIOSH draft method 5525 provides flexibility, enhanced sensitivity and specificity, powerful resolution, and very small compound-to-compound variability in the UV254, resulting in a more reliable identification and quantification of aliphatic isocyanates. PMID- 12094929 TI - Low-flow active and passive sampling of VOCs using thermal desorption tubes: theory and application at an offset printing facility. AB - While air sampling techniques using adsorbent-based collection, thermal desorption and chromatographic analysis have found a niche in ambient air sampling, occupational applications have been more limited. This paper evaluates the use of thermal desorption techniques for low flow active and passive sampling configurations which allow conveniently long duration sampling in occupational settings and other high concentration environments. The use of an orifice enables flows as low as 0.5 ml min(-1) and sampling periods up to several days without significant biases. A model is used to predict sampling rates of a passive sampler encompassing an orifice, a void space, glass wool, and the adsorbent. Laboratory and field tests conducted at a commercial offset printing facility, which contained a variety of volatile organic compounds (primarily aromatic but also a few chlorinated and terpene compounds at levels from 1 to 67,000 microg m( 3)), are used to evaluate the approach. Tenax GR and Carbosieve SIII, both singly and together, were employed as adsorbents. Side-by-side tests comparing high flow, low flow and passive samplers show excellent agreement and high linearity (r = 0.95) for concentrations spanning nearly five orders of magnitude. Active samplers were tested at flows as low as 0.5 ml min(-1), compared to typical flows up to 40 ml min(-1). Passive samplers demonstrated a linear range and agreement with predictions for adsorbate loadings from approximately 1 ng to nearly 10 microg. Using a chemical mass balance receptor model, concentrations in the facility were apportioned to solvents, inks and other indoor and outdoor sources. Overall, the use of low flow active and passive sampling approaches employing thermal desorption techniques provides good performance and tremendous flexibility that facilitates use in many applications, including workplace settings. PMID- 12094930 TI - Novel integrative passive samplers for the long-term monitoring of semivolatile organic air pollutants. AB - Two types of passive sampler were developed for the long-term monitoring of semivolatile organic compounds (SOCs) in air. They consist of poly(dimethylsiloxane) (PDMS)-coated stir bars (type A) or silicone tubing (type B), acting as a solid receiving medium, enclosed in a heat-sealed low-density polyethylene (LDPE) membrane. These samplers combine the advantages of integrative passive sampling with those of analysing accumulated analytes by thermodesorption-GC-MS, whilst avoiding the use of solvents and expensive sample preparation and cleanup steps. The performance of these samplers was investigated for the integrative sampling of SOCs, including alpha- and gamma hexachlorocyclohexanes, hexachlorobenzene, 2,4,4'-trichlorobiphenyl, 2,2',5,5' tetrachlorobiphenyl and fluoranthene, in laboratory exposure experiments under controlled conditions. For both types of sampler, the uptake of all the analytes investigated was linear over an exposure period of 15 days. The sampling rates calculated ranged from 70 to 320 ml h(-1) (sampler A) and 630 to 4300 ml h(-1) (sampler B). The passive samplers are able to detect low time-weighted average air concentrations in the pg m(-3) range. The small, robust and inexpensive sampling devices were tested successfully for the long-term air monitoring of semivolatile organic pollutants in a polluted area over an exposure period of up to 28 days. PMID- 12094931 TI - Evaluation of PCR primers and PCR conditions for specific detection of common airborne fungi. AB - We examined the selectivity of 53 sets of primers for environmental monitoring of indoor air quality. Thirty-six fungal strains, representing 26 species from 14 genera of commonly occurring fungi, and 16 different bacterial strains, representing both gram-negative and gram-positive species, were included in the experiment. We verified the specificity of 28 of the 53 sets of primers, which were classified as universal fungal, universal bacterial, group or species specific. The PCR conditions required for optimal specificity were also determined. These results can serve as a guide for the step-wise PCR-based detection and identification of airborne fungi commonly found in indoor environments. PMID- 12094932 TI - Toxic equivalency factors for PAH and their applicability in shellfish pollution monitoring studies. AB - Fish and shellfish are exposed to a wide range of polycyclic aromatic hydrocarbons (PAH) following oil spills at sea, and can become contaminated as a result. Finfish have a more effective mixed-function oxidase enzyme system than shellfish, and are therefore able to metabolise and excrete PAH more effectively than the invertebrates. Thus, contamination by high-molecular weight PAH, including those with carcinogenic potential and so of concern with regard to human consumers, is therefore usually observed in shellfish, and particularly in bivalve molluscs. Oil spills are not the sole source of PAH, however, as parent compounds are also generated by a wide range of combustion processes. In this paper, consideration is given to monitoring data gathered following recent oil spills (both of crude oil and diesel fuel), alongside data from other studies. These include studies conducted around a former gasworks site and downstream of an aluminium smelter in the UK, and from mussel monitoring studies undertaken in the UK and the USA (including the Exxon Valdez oil spill and the National Status and Trends programme), and in other countries in Europe. For comparative purposes the PAH concentrations are summed and also expressed as benzo[a]pyrene equivalents, their relative concentrations being weighted in relation to the carcinogenic potential of individual PAH compounds using toxic equivalency factors (TEF). Our aim was to assess the utility of this approach in fishery resource monitoring and control following oil spills. Certainly this approach seems useful from the data assessed in this study. and the relative ranking of the various studies seems to reflect the relative degree of concern for human consumers due to the differing contamination sources. As a simple tool for control purposes it is equally applicable to PAH derived from oil spills, and from industrial and combustion sources. PMID- 12094933 TI - Enantiomer ratios, patterns and levels of toxaphene congeners in human milk from Germany. AB - High resolution gas chromatography (HRGC) coupled to quadrupole negative ion chemical ionization (NICI) mass spectrometry (MS) was used to investigate congener patterns, levels and enantiomer distribution of selected toxaphene congeners (#26, #41, #44, #50, #63, B7-1453) in human milk from Germany. #50 and #26 were the most abundant congeners. Furthermore, the identification of B7-1453, B8-1412, #41, #42, #44 and #63 was possible. Levels for the sum of #26, #41, #44 and #50 ranged from 7 to 24 microg kg(-1) milk fat and contributed between 2 and 9% to the total burden of investigated organochlorines such as selected compounds of the chlordane group, HCH, DDT and polychlorinated biphenyls (PCB). Capillaries coated with heptakis-(2,3,6-O-tert-butyldimethylsilyl)-beta-cyclodextrin (TBDMS CD) or octakis-(2,3,6-tri-O-ethyl)-gamma-cyclodextrin (TEG-CD) were used for the enantioselective separations. Enantiomer ratios (ER) and enantiomer fractions (EF) of the abundant toxaphene congeners #26 and #50 as well as of B7-1453, #41 and #63 were determined. Greatest deviations from a racemic composition in individual human milk samples were found for #41 (1.54-2.37), #50 (1.37-1.72) and #63 (0.53-0.71) whereas ERs for #26 were close to 1. Compared to wildlife biota such as fish and raptors ER changes were more pronounced in human milk but comparable to human adipose tissue. PMID- 12094934 TI - Monitoring dioxins and furans in the vicinity of an old municipal waste incinerator after pronounced reductions of the atmospheric emissions. AB - In order to get an overall picture of the environmental impact of an old municipal solid waste incinerator (MSWI) from S. Adria del Besos (Barcelona, Catalonia, Spain), a monitoring program addressed at determining the levels of a number of pollutants in the vicinity of the facility was initiated in 1998. In March 1999, an adaptation was carried out due to EU legislation on pollutant emissions from the stack. As a result, emissions of polychlorinated dibenzo-p dioxins (PCDDs) and dibenzofurans (PCDFs) were notably reduced, and a significant (30%) decrease was found in the levels of PCDD/F in herbage samples collected in the vicinity of the MSWI. In March 2001, herbage samples were collected at the same sampling points and the PCDD/F levels measured again. The current PCDD/F concentrations range from 0.23 to 1.43 ng I-TEQ kg(-1) (dry matter), with median and mean values of 0.58 and 0.66 ng I-TEQ kg(-1) (dry matter), respectively, while in the 2000 survey the PCDD/F concentrations ranged from 0.22 to 1.20 ng I TEQ kg(-1) (dry matter), with median and mean values of 0.57 and 0.61 ng I-TEQ kg(-1) (dry matter), respectively. Although the current PCDD/F concentrations in herbage samples are comparable to those found in recent surveys carried out in various places of Catalonia, an exhaustive evaluation of the data, including principal component analysis, indicates that other emission sources of PCDD/Fs also have a notable environmental impact on the area under direct influence of the MSWI. PMID- 12094935 TI - Sources of lead and zinc associated with metal smelting activities in the Trail area, British Columbia, Canada. AB - The spatial distribution and deposition of lead and zinc emitted from the Trail smelter, British Columbia, Canada, was studied by strategically locating moss bags in the area surrounding the smelter and monitoring the deposition of elements every three months. A combined diffusion/distribution model was applied to estimate the relative contribution of stack-emitted material and material emitted from the secondary sources (e.g., wind-blown dust from ore/slag storage piles, uncovered transportation/trucking of ore, and historical dust). The results indicate that secondary sources are the major contributor of lead and zinc deposited within a short distance from the smelter. Gradually, the stack emissions become the main source of Pb and Zn at greater distances from the smelter. Typical material originating from each source was characterized by SEM/EDX, which indicated a marked difference in their morphology and chemical composition. PMID- 12094937 TI - Uptake of cadmium by the invasive perennial weeds Ranunculus repens and Geranium robertianum under laboratory conditions. AB - The objective of this study was to compare the accumulation and partitioning of cadmium (Cd) in a fibrous versus a tap root weed, Ranunculus repens and Geranium robertianum respectively. To meet this objective, we compared the accumulation by and the partitioning of Cd in R repens versus G. robertianum grown in soils spiked with 0.015 grams of Cd for a period of three weeks. The rate of Cd uptake was also compared by following the fate of 109Cd within the root, stem and leaf of the two weeds. Prior to Cd exposure, leaf and stem of control R. repens contained significantly greater amounts of Cd as compared to G. robertianum, whereas Cd concentrations in roots of the control plants for the two species were not significantly different (p > 0.05, student's t-test). Post Cd exposure the two species contained similar amounts of Cd in leaf and stem, however, roots of R. repens contained almost two-fold the amounts of Cd as compared to G. robertianum. Comparison of k (h(-1), rate of 109Cd uptake) for stem, leaf and root of the two species indicated that G. robertianum accumulated 109Cd over the first 24-48 h at a faster rate as compared to R. repens. For both species and all three organs, maximum accumulation of 109Cd occurred within the first 24-48 h. Our findings indicate that the fate of Cd within these two species is quite different with the fibrous root of R. repens serving to accumulate and store Cd whereas in G. robertianum, Cd is rapidly taken up and tends to be accumulated within its leaf. PMID- 12094936 TI - Should foliar cadmium concentrations be expressed on a dry weight or dry ash weight basis? AB - Foliar analysis is a valuable tool for evaluating the pollution status of forests. However, the use of foliar diagnosis in large-scale surveys is a complicated process owing to the high variability within the crown. The method used to express foliar concentrations has often been found to diminish the variability. The effect of the method used to express element concentrations on the spatial variability of cadmium (Cd) in the leaves of crack willow (Salix fragilis L.) was investigated by sampling the leaves of one willow at 292 locations in the crown, each sampling location having a volume of 0.027 m3 (0.3 m x 0.3 m x 0.3 m). Cadmium showed a distinct spatial trend in the crown of the tree. Concentrations as low as 2.4 mg kg(-1) dry weight (DW) or 23.1 mg kg(-1) dry ash weight (DAW) were obtained in the top of the crown, and 10.6 mg kg(-1) DW or 73.0 mg kg(-1) DAW in the bottom of the crown. The lower relative standard deviation and weaker correlation with the sampling height support the use of DAW in large-scale surveys especially. The lower variability of the DAW Cd concentration makes this variable less sensitive to fluctuations caused by differences in growing conditions and sampling methodology. However, the majority of publications in this field report metal concentrations on a DW basis. Therefore, the restrictions set on the use of results expressed on a DAW basis in large-scale surveys of foliar metal concentrations have to be offset against the advantages offered by a reduction of the variability in metal concentrations. PMID- 12094938 TI - Survey of four marine antifoulant constituents (copper, zinc, diuron and Irgarol 1051) in two UK estuaries. AB - A field survey of antifoulant concentrations was undertaken in two UK estuaries (Hamble and Orwell) in 1998 and 1999. The two locations offered variations in physical aspects (Orwell estuary being significantly larger than the Hamble) as well as differences in boat densities (Hamble having almost twice as many vessels moored in the estuary and marinas). Samples were analysed for copper, zinc, diuron and Irgarol 1051, and were collected in summer and winter in order to identify potential seasonal variations in concentrations. The effect that different marina types (e.g. locked marina, one located in a natural inlet and pontooned ones in the open estuary) had on antifoulant concentrations were also investigated. Concentrations of the organic booster biocides, diuron and Irgarol 1051 in the marinas and estuaries were mainly influenced by leaching from antifoulant paints applied to the hulls of leisure craft, and so levels reflected the number of vessels present in the water. As a consequence significantly higher concentrations were found in marinas (up to ca. 900 ng l(-1) for diuron and 240 ng l(-1) for Irgarol 1051) compared with estuaries (up to ca. 400 ng l(-1) for diuron and 100 ng l(-1) for Irgarol 1051) and in summer compared with winter. Sediment concentrations of Irgarol 1051 and diuron were rarely detectable other than in the marinas where high concentrations were detected near slipways assumed to be derived from washed off paint chips. Dissolved concentration profiles for copper and zinc in the estuaries and marinas were different from those for the organic booster biocides partly because other sources of these metals contributed to estuarine and marina loads. In particular, riverine loads and inputs from sacrificial anodes attached to leisure craft, exhibited a major influence of estuarine levels of zinc. Consequently, only in the Hamble estuary for copper was there a clear distinction between summer (typically 3-4 microg l(-1)) and winter dissolved values (typically 1-2 microg l(-1)) that could be largely attributable to the leaching of antifoulant paints. Sediment concentrations for both metals were similar for both estuaries, with little variation between winter and summer values (Zn ranging from 28 to 614 mg kg(-1) and Cu from 6 to 1016 mg kg(-1)) as with the organic booster biocides highest levels were measured at the base of slipways in marinas. PMID- 12094939 TI - Organotin compounds in the marine environment of the Bay of Piran, northern Adriatic Sea. AB - In the year 2000, organotin pollution was investigated in the Bay of Piran, Slovenia, at the northern extremity of the Adriatic Sea. Gas chromatography-mass spectrometry with single ion monitoring GC/MS-SIM was applied for speciation analysis of pentylated organotin compounds in water and mussels. Sampling of marine waters was performed in summer and autumn at 8 locations. Mussels Mytilus galloprovincialis were collected only in summer at 3 locations. The highest concentrations of tributyltin (TBT) in marine water ranged from 500 to 630 ng L( 1) (as Sn) in summer and from 180 to 230 ng L(-1) (as Sn) in autumn. TBT concentration in mussels varied from 500 to 3500 ng g(-1) dry weight (d.w.) (as Sn). The study indicated remarkable butyltin pollution of the Slovenian coastline. Accuracy of the analytical procedure was verified by BCR CRM 477. PMID- 12094940 TI - Distribution of HCH and DDT in the coastal marine environment of Mumbai, India. AB - Organochlorine pesticides (OCPs) in sediment, water and biota samples from the coastal marine environment of Mumbai were analysed to elucidate their distribution in various environmental compartments. HCH isomers, dichlorodiphenyltrichloroethane (DDT) and its metabolites were the predominantly identified compounds in all the samples. High ratios of DDT to dichlorodiphenyldichloroethylene (DDE) were found in seawater samples. This indicates the presence of a significant source of DDT in this region. In sediment samples, the gamma-isomer contributed almost 55% to the total HCH, indicating a high affinity of the gamma-isomer towards the sediment. The levels of organochlorines in fish obtained from this region were found to be lower than the levels of organochlorines in fish in temperate regions. PMID- 12094941 TI - Transport and fate of organochlorine pesticides in the River Wuchuan, southeast China. AB - Persistent organic pollutants (POPs) such as chlorinated pesticides are of global concern due to their widespread occurrence, persistence, bioaccumulation and toxicity to animals and human. This paper summarises recent research on 18 chlorinated pesticides in an important catchment in China, by determining their concentrations and behaviour in water, sediment, soil and plants. The concentrations of the total pesticides were in the ranges 187-893 ng l(-1) in river water, 8.53-210 ng g(-1) dry weight in soil, 2.66-13.45 ng g(-1) dry weight in river sediment, and 651-2823 ng g(-1) dry weight in plants. The predominance of beta-HCH as the major isomer of HCHs in all water, soil, sediment and plant samples was clearly observed, due to beta-HCH's resistance to biodegradation. On average beta-HCH accounted for 44%, 53%, 50%, and 46% of the total HCH concentration in water, soil, sediment and plant, respectively. Of the DDTs, DDE accounted for 48%, 43%, 53%, 55% of the total DDT, which suggested that DDT had been transformed to its metabolites, DDE and DDD, of which DDE was the more stable. The chlorinated pesticide levels in the River Wuchuan were generally below the guideline values in China, but some sites displayed levels in excess of EC Environmental Quality Standards for HCHs and DDTs. The results therefore provide important information on the current contamination status of a key agricultural watershed in China, and point to the need for urgent actions to evaluate the long-term fate and toxicity of such persistent compounds and an appropriate remediation strategy. PMID- 12094942 TI - Development of an enzyme-linked immunosorbent assay for screening contamination by chlorophenols in environmental waters. AB - The development of an immunoassay for screening contamination by chlorophenols is presented. Two haptens were synthesized and conjugated to immunizing proteins to raise rabbit polyclonal antibodies. The antibody-coated format (direct) gave better sensitivity than the conjugate-coated format (indirect) if 2,4,6 trichlorophenol is used as target analyte. The measurement range was 86.4 microg l(-1) to 0.7 microg l(-1), with an average I50 of 7.8 microg l(-1) and a detection limit of 0.2 microg l(-1). The assay detects the presence of trichloropyridinol and other chlorophenols such as di-, tetra- and pentachlorophenols constituting thus a suitable tool for the early warning of the presence of such family contaminants. The optimized method permits the detection of the most important chlorophenols in a fast and reproducible way using no more than one antibody and a single assay. The results achieved with water samples spiked with different chlorophenols fit with a multiple linear regression model when expressing the total concentration of chlorophenols as equivalent of 2,4,6 trichlorophenol (P < 0.01), demonstrating the usefulness of the assay as a screening tool to detect contamination by chlorophenols. PMID- 12094943 TI - Identification of new photoreduction products of pentachlorophenol. AB - Pentachlorophenol (PCP), a fungicide and insecticide, leaves behind considerable residue in the environment in the form of unreacted molecule, metabolites and photoreduction products. Most of the previously reported photoproducts have resulted from dechlorination of the parent molecule. We have shown that photoreduction of PCP also proceeds by photoreduction of the parent molecule at carbons 2 and 4. These products were identified and characterized as 2-dehydro 2,3,4,5,6-pentachlorocyclohexanone (II) and 2,3,4,5,6-pentachlorocyclohexanone (IV). Both of them display keto-enol tautomerism. Identification and characterization of new photoproducts of PCP is significant for determining its overall environmental impact in the ecosystem. PMID- 12094944 TI - Dechlorination of pentachlorophenol in supercritical carbon dioxide with zero valent palladium-magnesium bimetallic mixture. AB - A stream of substrate pentachlorophenol [PCP, 5 mg min(-1) in water-methanol (1 + 4, v/v)] was merged with 1.5 ml min(-1) of supercritical carbon dioxide (scCO2) and delivered to a reactor column (25 cm x 1 cm) of zero-valent palladium magnesium mixture. The resulting dechlorinations, although very efficient, were not quantitative. For continuous operation at 400 degrees C for 6 h, phenol was the principal product, with lesser quantities of methylated products and only traces of chlorinated products (principally monochlorinated species). PCP deoxygenation was not observed and ring methylation was decreased relative to analogous reactions in hydroxylic organic solvent. With time, the reactor column slowly lost dechlorination activity. Reducing the loading of Pd0 on Mg0 from 2% to 1% (w/w) apparently did not change the course of the reaction; however, the dechlorination capacity was decreased correspondingly. None the less, over 6 h or 5 h of continued operation, the dechlorination efficiency was 0.995 for the 2% (w/w) loading of Pd0 on Mg0 and 0.984 for the 1% (w/w) loading. PMID- 12094945 TI - Turbidimetric determination of chloride in different types of water using a single sequential injection analysis system. AB - A sequential injection analysis system for the turbidimetric determination of chloride in different types of water is proposed. The determination is based on the reaction of chloride with silver ions and the subsequent measurement of the turbidity caused by silver chloride precipitation. In this method, the use of toxic reagents, such as mercury thiocyanate, commonly employed in most spectrophotometric techniques for chloride determination, is avoided. The main feature of the developed system is the use of a single configuration to carry out the determination over a wide concentration range (2-400 mg L(-1)) by changing only the aspirated sample volume. This characteristic allows the determination of chloride in ground, surface and wastewaters using the same manifold. In addition, a considerable saving of precipitating reagent is achieved due to non-continuous consumption. The results obtained with the developed system were statistically indistinguishable from those of the potentiometric titration reference method. Relative standard deviations for ten consecutive injections were lower than 3.7%, with a sampling frequency of between 55 and 57 determinations per hour. PMID- 12094946 TI - Regeneration of granular activated carbon loaded with explosives. AB - Granular activated carbon (GAC) loaded with trinitrotoluene (TNT) and nitrobenzene (NB) were subjected to different atmospheres, such as nitrogen, oxygen, air, steam and an air-steam mixture, at varying temperatures and for different time intervals for regeneration. The air-steam mixture proved to be the best regenerating media for the explosive-loaded GACs. The regeneration of the activated carbon was found to be above 85% for TNT and above 90% for NB loaded carbons. PMID- 12094947 TI - Current strategies in nitrite detection and their application to field analysis. AB - The various analytical protocols that have been developed to aid the measurement of nitrite within environmental samples have been critically appraised and their applicability to field measurement assessed. The present communication presents a perspective on current techniques and technologies encompassing spectroscopic, electrochemical and chromatographic methodologies and highlights those that are liable to emerge in the near future. Commercial devices have been included and where appropriate the advantages and limitations posed by their operation within field contexts have been described. PMID- 12094948 TI - Grading in ovarian cancer. PMID- 12094949 TI - Radioimmunodetection of cervical carcinoma xenografts with (111)In-labeled MAb Cx 99 detected by a hand-held gamma detector. AB - PURPOSE: To establish a radioimmunodetection (RAID) system for localization of cervical cancer by labeling 111-indium ((111)In) to a monoclonal antibody against cytokeratin 19 (MAb Cx-99), and detecting it with a hand-held gamma detector in an animal model. METHODS: MAb Cx-99 was labeled with 111-Indium by the DTPA chelating method. From the second day to the seventh day after injection of this immunoconjugate into athymic nude mice bearing cervical cancer cell line CC7T xenografts, the biodistribution ratios of tumor and non-tumor radioactivity were detected by a hand-held gamma detector. Data were also correlated with the data detected by the conventional gamma counter. RESULTS: The labeling efficiency of this (111)In-labeled MAb Cx-99 and (111)In-labeled MOPC was 91.6% and 95.5%, respectively. After injection, the liver, kidney and lung were initially noticed to have high radioactivity, but the localization of tumor/tissue ratios increased progressively as time passed, indicating the effect of delayed detection for distinguishing tumor from non-tumor tissues. Except for the spleen, the range of tumor/tissue ratios was 1.18-32.7 and 1.14-39.35 for the fourth day and the seventh day, respectively. The tumor/spleen ratio remained low until the seventh day after injection, thus indicating that the spleen might have a different excretion rate. CONCLUSION: This study indicated the feasibility of a hand-held detection system in the localization of cervical cancer after injection of (111)In-labeled MAb Cx-99. The effect of delayed detection was obvious by the decreasing high bindings in the liver, spleen and kidney, with the applicable detection time being four to seven days after injection. PMID- 12094950 TI - Renin-producing serous cystoadenocarcinoma of the ovary: a case report. AB - BACKGROUND: Only a few renin-producing ovarian tumors have been reported, and most such ovarian tumors have been sex cord/stromal tumors. Renin-producing ovarian epithelial tumors are quite rare. CASE: A 46-year-old woman presented with hypertension and hypokalemia. Examinations of the patient revealed elevated plasma renin activity, hyperaldosteronism and a pelvic mass. Subsequently, a right ovarian tumor mass was resected. Microscopic observation of the tumor revealed a well-differentiated serous cystadenocarcinoma. Immediately after surgery, blood pressure, serum potassium, plasma renin activity and plasma aldosterone levels returned to normal ranges. RT-PCR analysis and immunohistochemical staining of this tumor indicated that it was producing renin. CONCLUSION: This is the first report of a renin-producing ovarian serous cystadenocarcinoma. PMID- 12094951 TI - Familial and hormonal risk factors for papillary serous uterine cancer. AB - OBJECTIVES: To identify genetic and non-genetic risk factors for papillary serous uterine cancer. METHODS: A case-control study was conducted. Case women with papillary serous uterine cancer were compared with two control groups: 1) women with endometrioid uterine cancer and 2) healthy women with no past history of cancer. Cases and controls were matched for age (within two years) and ethnic group. All study subjects completed a questionnaire addressing family history. The cases and healthy controls were assessed for factors associated with estrogen exposure. RESULTS: The risks of breast cancer (RR 1.84, CI 1.03-3.31) and of prostate cancer (RR 2.21, CI 0.77-6.37) were higher among the relatives of patients with papillary serous uterine cancer, than among relatives of those with endometrioid uterine cancer. Other significant risk factors included weight at 18 years (p = 0.04) and the use of estrogen replacement therapy (p = 0.04). CONCLUSION: Relatives of women with papillary serous cancer of the uterus had an increased risk of breast and prostate cancer. Hormonal exposure also increases the risk for this cancer. These findings suggest that predisposing genetic factors, possibly related to hormone metabolism, may be common to the three forms of cancer. PMID- 12094952 TI - Sclerosis of gross cysts of the breast: a three-year study. AB - Breast cysts can be separated into two types: Type I cyst with a lining epithelium which shows apocrine metaplasia, and Type II cyst with an epithelium which is markedly attenuated or absent. The risk of subsequent breast cancer among patients with Type I cysts can be up to 4. The standard treatment is fine needle aspiration, but 20% of the cysts recur. Pharmacological treatment has been tried, which reduces size and volume, but has side-effects and a high recurrence rate post-treatment occurs. The objectives of this prospective study were to sclerose the cyst, induce its regression and prevent or reduce recurrence rate, with the administration of a sclerosing solution (Sclerovein) within the cyst post-aspiration. Fifty-seven patients were followed in the study, 37 with Type I cysts and 20 with Type II cysts. At the end of six months all patients with Type II cysts had no detectable cyst. On the other hand, two patients still had a residual Type I cyst. At the end of three years our recurrence rate appears to be less than 2%, with one patient with a possible recurrence. No significant side effects were observed. The use of Sclerovein is a simple and safe alternative in the treatment of recurring cysts. PMID- 12094953 TI - Primary malignant melanoma of the vagina: case report and review of literature. AB - Primary vaginal malignant melanoma is rare, with < 250 reported cases to date. It accounts for < 1% of all melanomas in women, < 10% of all female genital tract melanomas, and < 3% of all vaginal malignancies. Its clinical behavior is notoriously more aggressive than that of cutaneous and vulvar melanoma, with a 5 year survival rate ranging from 5% to 25%. Tumor size is the strongest predictor of survival, whereas tumor thickness is a weak predictor of survival. A case of F.I.G.O. stage I vaginal melanoma encircling and embracing the entire circumference of the middle third of the vagina is described. Tumor size and thickness were 6 cm and 9 mm, respectively. The patient was treated by wide local excision and pelvic radiotherapy, and to date, 11 months after surgery, she is alive and with no evidence of disease. It is concluded that wide local excision followed by pelvic radiotherapy is an appropriate treatment for melanoma confined to the vagina. PMID- 12094954 TI - Desmoplastic small round cell tumor: report of a case presenting as a primary ovarian neoplasm. AB - BACKGROUND: Desmoplastic small round cell tumor (DSRCT) is an intra-abdominal malignancy that typically has extensive peritoneal spread at the time of diagnosis. We report a case of DSRCT with involvement of the ovary and omentum as well as an elevated CA-125 level at presentation. CASE: A 23-year-old female presented to another institution with a pelvic mass and a CA-125 level of 140 U/ml. During tumor reductive surgery the right ovary, omentum, and liver were found to be involved. Initial histologic examination favored an undifferentiated small cell carcinoma of the ovary. The patient received two cycles of Taxol and cisplatin chemotherapy and was referred to the University of Texas M. D. Anderson Cancer Center. Upon review of the pathology material at the time of the referral, a diagnosis of DSRCT was made. Despite two additional cycles of chemotherapy, the tumor progressed, and the patient returned home. CONCLUSION: DSRCT may mimic an ovarian primary tumor by presenting with involvement of the ovary and an elevated CA-125 level, and should be included in the differential diagnosis of ovarian neoplasms in young patients. PMID- 12094955 TI - Localized distribution of human papillomavirus genotypes in the uterine cervix. AB - INTRODUCTION: The localization and distribution of single or multiple HPV genotypes in the uterine cervix has not been studied thus far. The present study was undertaken to determine whether single or multiple HPV genotypes detected in cervical smears originate from a single (dysplastic) area, or from different areas (dysplastic or normal) of the uterine cervix. METHODS: Of eight patients with moderate or severe dysplasia, 31 colposcopically guided biopsies of different dysplastic lesions of the uterine cervix, as well as of normal epithelium were investigated. A highly sensitive, broad spectrum, short fragment polymerase chain reaction (SPF-10 PCR) HPV detection method in combination with a line probe assay (LiPA) for simultaneous genotyping was used. RESULTS: In the uterine cervix of four of the eight patients, multiple HPV genotypes were detected. These multiple HPV genotypes were detected in different biopsies as well as within a single biopsy. In three patients, all with carcinoma in situ or microinvasive carcinoma, only a single HPV genotype, HPV 16, was found all over the cervix including in the normal epithelium. CONCLUSION: Different HPV genotypes can be detected in different dysplastic lesions as well as within single lesions, especially in patients with severe dysplasia. The severity of the lesion may possibly have a relation with the distribution of the HPV genotypes. The low number of patients and biopsies does not allow definite conclusions. However, the impact of these findings on the outcome of screening and vaccination programs remains to be elucidated. PMID- 12094956 TI - Outcome of pregnancies after cold-knife conization of the uterine cervix during pregnancy. AB - OBJECTIVE: To determine the incidence of cervical intraepithelial neoplasia (CIN) in pregnancy and to determine the outcome of pregnancies in women treated by cold knife conization during pregnancy. METHODS: The authors retrospectively studied the cases of 19,807 pregnant patients, who presented to the 1st Department of Obstetrics and Gynecology Semmelweis University Faculty of Medicine between January 1, 1993 and December 31, 1997. Of these, there were 1,513 spontaneous abortions, 6,170 artificial abortions and 12,124 deliveries. Cytological evidence of a high-grade squamous intraepithelial lesion (LGSIL) was found in 48 pregnant patients. A low-grade squamous intraepithelial lesion (LGSIL) was found in 55 pregnant patients. All pregnant patients with HGSIL including those with satisfactory and nonsatisfactory colposcopical examinations underwent diagnostic cold knife conization during pregnancy in the second trimester. A gynecologic pathologist (Dr. Zs. Csapo) reviewed the histopathological sections. RESULTS: Overall, 44 patients had CIN, three patients had microinvasive carcinoma (FIGO stage I/A1), and one patient had neither CIN nor invasive carcinoma in the HGSIL group. Among the 48 pregnant patients with HGSIL, there were two spontaneous abortions occurring during the 21st and the 22nd gestational weeks respectively, so the pregnancy loss rate was 4.2%. Forty patients (83%) delivered at term and six patients (12.5%) at preterm. Twenty-four patients (52%) delivered vaginally and 22 (48%) by cesarean section. Comparing this data with the control group consisting of pregnant patients who had LGSIL, the difference between the two group as regards the total number of adverse pregnancy outcome cases was not significant. In the three patients with microinvasive cervical carcinoma (FIGO stage I/A1), elctive cesarean section was performed, followed by an immediate abdominal hysterectomy at term. CONCLUSION: The incidence of CIN in pregnancy was 0.22%. The incidence of microinvasive cervical carcinoma was 0.015%. Pregnant patients with CIN who underwent cold knife conization during pregnancy were not at increased risk of adverse pregnancy outcome, however they were at increased risk of cesarean delivery. PMID- 12094957 TI - Papnet-assisted cytological diagnosis intensifies the already marked variability among cytological laboratories. AB - OBJECTIVE: The main objective was to assess the sensitivity, specificity and reliability of PAPNET-assisted diagnosis in comparison with conventional screening. SETTING: Seven Italian and one English University or Research Institutes, and a random sample of an other 20 Italian Laboratories of the Italian National Health Service (INHS) provided the cervical smears. METHODS: During the training phase every center examined in rotation four sets of slides for a total of 300 representative slides. Afterwards, 900 "positive" slides were added to the 3,100 slides which were collected consecutively without any selection or exclusion. The eight main centers were divided into four couples and each couple of centers examined 775 slides with the PAPNET system, "blindly" to the original diagnosis. An expert cytopathologist (M.A.) of the National Institute of Health (NIH) reassessed 40% of the slides with an original negative diagnosis to evaluate the false negative rate. Two expert NIH cytopathologists (M.A., G.M.) re-examined all slides where a disagreement had been observed between the original and one or both of the study diagnoses. The main analyses concerned the following three main categories: WNL and unsatisfactory for evaluation; ASCUS, AGUS and LSIL; HSIL and carcinoma. A special algorithm was devised to define the reference diagnosis for sensitivity and specificity assessment. RESULTS: Laboratories, even belonging to the same couple, classified as "no review" a very different proportion of slides ranging from 35% to 74%. The index of kappa agreement between the members of couples examining the same sets of slides was low or very low, ranging from 0.30 to 0.03. The sensitivity of the review classification was particularly low in some laboratories. Surprisingly, only a small correlation was observed between the sensitivity of the review classification and the proportion of slides classified as "review". The "tentative" diagnosis on PAPNET tiles of the "review" slides was almost as reliable as the microscopic diagnosis. In the overall performance, there were many significant differences among the eight laboratories. The best laboratory had a sensitivity of 95% and a specificity of 96%. At least three laboratories displayed unacceptably low sensitivity and one a very low specificity. CONCLUSION: Altogether these results seem to confirm that there are wide differences among cytological laboratories per se, and that these differences are intensified by the use of an instrument like PAPNET. The huge variation in performance may be explained by differences in basic skills and by different training, but it is difficult to understand exactly what could have been done to reduce it. PMID- 12094958 TI - Identification of high risk patients with endometrial carcinoma. Prognostic assessment of endometrial cancer. AB - OBJECTIVES: To verify the importance of DNA ploidy on clinical outcome in endometrial carcinoma and to investigate whether the prognostic information obtained by this variable is independent from other clinical-pathologic features. MATERIALS AND METHODS: Univariate and multivariate analysis of clinical and pathologic prognostic factors obtained from 203 consecutive cases of endometrial cancer, that had been surgically treated in our hospital, were performed. RESULTS: Significant prognostic factors according to the Kaplan-Meier method were age at the time of diagnosis, grade of differentiation, peritoneal cytology, node involvement, vascular invasion, myometrial infiltration and ploidy. At multivariate analysis only DNA ploidy resulted to be an independent variable. CONCLUSIONS: In our analysis DNA content is the only parameter which preserved prognostic significance in multivariate analysis. PMID- 12094959 TI - Different enzyme activities of sialyltransferases in gynecological cancer cell lines. AB - PURPOSE: Due to rarity of a relationship between sialyltransferase enzyme activities and gynecological cancers, we arranged the study to evaluate sialyltransferase enzyme activity in the various kinds of gynecological cancer cell lines. METHODS: Ten cell lines from various kinds of gynecological cancers and two cell lines from normal tissue were enrolled in this study. The activities of each subtype of sialyltransferases were detected using Gal beta1,3GalNAc acetyl-lactosamine)-Obz1 (acceptor for ST2,3Gal I), Gal beta1,3GlcNAc beta1,3Gal beta1,4GlcNAc (acceptor for ST2,3Gal III), Gal beta1,4GlcNAc (acceptor for ST2,3Gal IV), asialo-bovine submaxillary mucin (acceptor for ST2,6GalNAc I), asialo-fetuin (acceptor for STalpha2,6GalNAc II), and fetuin (acceptor for ST2,6GalNAc III), respectively. The amounts of sialic acids were measured using fluorescein-conjugated Sambucus nigra agglutinin (SNA) specific for alpha2,6 sialic acids and fluorescein-conjugated Maackia Amurensis agglutinin (MAA) specific for alpha2,3-sialic acids. RESULTS: The activities of two sialyltransferase subtypes, ST3Gal I & ST6GalNAc II, were significantly higher in nearly all cell lines. More specifically, cervical cancer cell line-ME 180, ovarian cancer cell line-ES-2, and choriocarcinoma cell line-BeWo showed high levels of ST3Gal I enzyme activity; all gynecological cancer cell lines except endometrial cancer cell line-RL95-2 had high levels of ST6GalNAc II enzyme activity when compared with a normal control cell line--fibroblast cell line (CCD 966Sk). Cell lines tested in this study have diverse levels of surface alpha2,6 sialic acid sugar chains (enhanced SNA binding) when compared with alpha2,3 sialic acid sugar chains (enhanced MAA binding) but we found that some cell lines such as Ca Ski (cervical cancer cell line), CC7T (cervical cancer cell line), PA I (ovarian cancer cell line), and BeWo showed significantly altered cell surface alpha2,6-sialic acid sugar chains. CONCLUSION: Increasing enzyme activity of ST3Gal I and ST6GalNAc II might be important in various kinds of gynecological cancers. More specifically, enhanced activity of sialyltransferases involving alpha2,6-sialic acid sugar chains might be more important in cancer development. Future studies will investigate whether the enzyme activity of these sialyltransferases can be helpful for clinical practice. PMID- 12094960 TI - Extramedullary hematopoiesis in the uterine isthmus: a case report and review of the literature. AB - Extramedullary hematopoiesis (EH) is the ectopic production of myeloid, erythroid and megakaryocytic elements. In postfetal life it usually occurs in conditions with hyperactive, depleted or infiltrated marrow; it is extremely rare in the genital tract. We report a case of EH in the uterine isthmus (UI) which was found incidentally in a 40-year-old patient who presented with a right ovarian cyst and a history of a right modified radical mastectomy for infiltrating invasive lobular carcinoma one year earlier. The patient underwent total abdominal hysterectomy with bilateral salpingo-oophorectomy. Microscopic examination of UI revealed foci of hematopoiesis, consisting of white blood cell precursors confirmed by positive chloroacetate-esterase staining. Bone marrow biospy showed diffuse infiltration secondary to breast cancer. Further work-up, including a bone scan, showed multiple metastases suggesting that the high degree of bone marrow infiltration by neoplasmatic cells had stimulated the EH. In conclusion, the unusual finding of EH in UI heralded widespread infiltration of bone marrow. PMID- 12094962 TI - Different types of hysterectomy in the radio-surgical treatment of early cervical cancer (FIGO Ib-IIa). AB - PURPOSE OF INVESTIGATION: Surgery and radiotherapy achieved equivalent results for FIGO stages Ib-IIa invasive cervical carcinoma. The integration of radiotherapy and surgery provided the same results for a selected series of patients without increasing the rate of complications. The aim of the study was to verify if, applying a radio-surgical protocol, the reduction of the surgery extension on the parametrium in one of two consecutive series might achieve the same results in terms of survival and recurrence rates with fewer complications. MATERIAL AND METHODS: We analysed actuarial survival (with >10-year follow-up), local control rates and morbidity of 390 patients who had different the kinds of surgery applied in the radio-surgical treatment protocol: Protocol A: brachytherapy plus type III radical hysterectomy vs Protocol B: brachytherapy plus type II radical hysterectomy. Patients were all included in an analysis of complications according to the French-Italian glossary. RESULTS: Analyses showed no differences in terms of survival, patterns of recurrences and onset time within the two protocols. Urinary complications were more frequent and severe in protocol A vs protocol B (G2: 26.5% vs. 6.1%; G3: 5.3% vs. 3.6%). CONCLUSION: Our study pointed out that the reduction of the surgery extension allowed the same overall survival and relapses with fewer complications particularly in terms of grade of severity. PMID- 12094961 TI - Evaluation of DNA mismatch repair system in cervical dysplasias and invasive carcinomas related to HPV infection. AB - The aim of this study was to answer the question whether the products of hMSH2 and hMLH1 genes take part in the mutation track of cervical carcinoma. METHODS: IgG1 monoclonal antibodies (Pharmingen) detecting epitopes characteristic of hMLH1 and hMSH2 were used in the present study. The value of the half quantitative H-score coefficient was calculated. Its threshold value was 0.4. Identification of 16 and 18 HPV types was performed by PCR. RESULTS: An intensified hMLH1 protein expression was observed both in the squamous epithelial carcinomas and cervical adenocarcinomas (H-score of 1.44 and 0.98, respectively) as compared to the control (H-score of 0.9). However, a decreased expression of hMSH2 protein was observed in the analysed cases of carcinoma (0.9 and 0.7) as compared to the control group (1.2). An intensified expression in G3 for hMLH1 and higher hMLH1 in comparison to hMSH2 was observed. CONCLUSIONS: 1. A considerable expression of hMLH1 and hMLH1 proteins was observed in the tissues with invasive cervical carcinoma not only within epithelial but also in stromal cells. 2. More intense expression of hMLH1 and hMSH2 was observed in invasive carcinomas and CIN than in the non-neoplastic cervical tissue lesions (erosion). 3. A stronger expression was observed for the hMLH1 than for the hMSH2 proteins- contrary to the cases of carcinomas of the uterine corpus and endometrial carcinoma. PMID- 12094963 TI - The significance of local cellular immune response of women 50 years of age and younger with endometrial carcinoma. AB - OBJECTIVE: Local cellular immune response and traditional histopathologic parameters in endometrial carcinoma patients 50 years old and younger were compared with those of patients older than 50 years of age. MATERIALS AND METHODS: We retrospectively compared the clinicopathologic factors and outcomes of 24 younger and 82 older women who were operated on for endometrioid type endometrial carcinoma at our institution. RESULTS: No significant difference was determined in survival with respect to age and menopausal status in patients with endometrial carcinoma. Younger and older age groups had a similar distribution of most pathologic features including myometrial invasion, cervical involvement, lymph node metastasis, vascular invasion, perivascular lymphocytic infiltrates and tumor infiltrating lymphocytes. While older women had higher-grade tumors, younger women had more frequent ovarian metastasis. Tumor infilrating lymphocytes were seen more frequently in the postmenopausal group than those of the premenopausal group. CONCLUSION: The distribution of local cellular immune response, most histopathologic parameters and survival were the same for younger and older women. PMID- 12094964 TI - Chlamydia trachomatis infection in cervical intraepithelial neoplasia and invasive carcinoma. AB - We analyzed 149 women (81 with cervical intraepithelial neoplasia and with invasive carcinoma of the cervix and 68--as a control group). The influence of Chlamydia trachomatis (Cht) infection into expression of EGFR, TGF-alpha, Ki 67, HPV 16 and 18 was examined. IS-PCR was used to measure the level of antibodies in the serum. We detected that chlamydial infection may cause cervical hypertrophy in women with and without cervical intraepithelial neoplasia and invasive carcinoma. Infections of both Cht and HPV correlate with high expession of Ki 67 in epithelium. Cht infection also increased the expression of HPV16 in CIN I. These results suggest that Cht infection modifies the activity of viruses. In our research we have confimed that Cht infection increases the expression of EGFR and TGF-alpha. These facts may explain variants other than the HPV-mechanism of cervical carcinogenesis. PMID- 12094965 TI - Malignant ovarian germ cell tumors: the KK Hospital experience. AB - Ovarian germ cell malignancies pose a therapeutic challenge especially amongst young patients. This is a retrospective review of 49 patients treated for such malignancies at KK Women's and Children's Hospital over a 13-year period. The relative proportion of such tumors was 6.2%. Age at presentation ranged from 14 to 51 years (mean 25.4 years). Forty-nine percent of tumors were immature teratomas and 81.6% had stage I disease. All patients had surgery initially and 67.3% required postoperative adjuvant chemotherapy. The patients were followed-up for one to 145 months (mean 51.6 months). All the 87.8% of patients on follow-up are alive and disease-free. There was one recurrence. Five patients had eight successful pregnancies, with no congenital anomalies. Mean duration when menstruation was resumed and regular was 2.5 and 3.5 months, respectively. With combination chemotherapy and conservative surgery, the outlook for patients is excellent, with emphasis on preservation of ovarian function and fertility. PMID- 12094966 TI - Ovarian cysts in postmenopausal tamoxifen-treated breast cancer patients with endometrial thickening detected by transvaginal sonography. AB - OBJECTIVE: To investigate the frequency of ovarian cysts in tamoxifen-treated postmenopausal breast cancer patients with endometrial thickening detected by transvaginal sonography. METHODS: Medical records and transvaginal sonographies of 38 postmenopausal women treated for breast cancer with adjuvant tamoxifen therapy who had undergone endometrial sampling due to abnormal endometrial thickness were reviewed retrospectively. RESULTS: During the study period five of 38 tamoxifen-treated postmenopausal patients (13.2%) had ovarian cysts. The mean tamoxifen treatment interval of the patients with an ovarian cyst was 22.4 +/- 18.4 months (p = 0.17). The mean endometrial thickness of the patients with an ovarian cyst was 12.6 +/- 5.9 mm (p = 0.17). Endometrial biopsy detected six cases of abnormal endometria, including endometrial carcinoma (n = 1), endometrial polyp (n = 1) and simple endometrial hyperplasia without atypia (n = 4). Three patients with ovarian cysts underwent laparatomy revealing simple cysts on histopathological examination. Two patients with ovarian cysts declined laparatomy and are currently under follow-up. CONCLUSION: Ovarian cysts a common side-effect of tamoxifen treatment in postmenopausal tamoxifen-treated breast cancer patients. Transvaginal sonography should be performed to detect any concomitant endometrial pathology. PMID- 12094967 TI - Expression of NM23 and tenascin in invasive ductal carcinomas of the breast. AB - OBJECTIVE: In this series of ductal carcinoma of the breast, immunoexpression of antimetastasis gene nm23 and tenascin was examined and the role in prognosis was investigated by correlation with the tumour grade and stage, and ER/PR immunoexpression. MATERIAL AND METHODS: In this study 27 ductal carcinomas of the breast were analysed for expression of tenascin and nm23 antimetastasis genes by immunohistochemistry. RESULTS: The results of our study revealed a statistically significant correlation between nm23-H1 immunoexpression and lymph node metastasis. We also found a statistically significant correlation between tenascin and nm23-H1 immunoexpression. Our results suggest that tenascin limits tumour spread. CONCLUSION(S): Antimetastasis gene expression can be used in predicting lymph node metastasis in ductal carcinomas of the breast. PMID- 12094968 TI - Abdominal skin metastasis of endometrial adenocarcinoma: case report. AB - Skin metastasis from endometrial adenocarcinoma in the radiotherapy field is reported. A 60-year-old woman with FIGO stage IB, grade 2 endometrial adenocarcinoma presented 24 months after initial surgery with skin metastasis located on the abdomen. PMID- 12094969 TI - The effect of voxel size on the accuracy of dose-volume histograms of prostate 125I seed implants. AB - Cumulative dose-volume histograms (DVH) are crucial in evaluating the quality of radioactive seed prostate implants. When calculating DVHs, the choice of voxel size is a compromise between computational speed (larger voxels) and accuracy (smaller voxels). We quantified the effect of voxel size on the accuracy of DVHs using an in-house computer program. The program was validated by comparison with a hand-calculated DVH for a single 0.4-U iodine-125 model 6711 seed. We used the program to find the voxel size required to obtain accurate DVHs of five iodine 125 prostate implant patients at our institution. One-millimeter cubes were sufficient to obtain DVHs that are accurate within 5% up to 200% of the prescription dose. For the five patient plans, we obtained good agreement with the VariSeed (version 6.7, Varian, USA) treatment planning software's DVH algorithm by using voxels with a sup-inf dimension equal to the spacing between successive transverse seed implant planes (5 mm). The volume that receives at least 200% of the target dose, V200, calculated by VariSeed was 30% to 43% larger than that calculated by our program with small voxels. The single-seed DVH calculated by VariSeed fell below the hand calculation by up to 50% at low doses (30 Gy), and above it by over 50% at high doses (>250 Gy). PMID- 12094970 TI - Direct aperture optimization: a turnkey solution for step-and-shoot IMRT. AB - IMRT treatment plans for step-and-shoot delivery have traditionally been produced through the optimization of intensity distributions (or maps) for each beam angle. The optimization step is followed by the application of a leaf-sequencing algorithm that translates each intensity map into a set of deliverable aperture shapes. In this article, we introduce an automated planning system in which we bypass the traditional intensity optimization, and instead directly optimize the shapes and the weights of the apertures. We call this approach "direct aperture optimization." This technique allows the user to specify the maximum number of apertures per beam direction, and hence provides significant control over the complexity of the treatment delivery. This is possible because the machine dependent delivery constraints imposed by the MLC are enforced within the aperture optimization algorithm rather than in a separate leaf-sequencing step. The leaf settings and the aperture intensities are optimized simultaneously using a simulated annealing algorithm. We have tested direct aperture optimization on a variety of patient cases using the EGS4/BEAM Monte Carlo package for our dose calculation engine. The results demonstrate that direct aperture optimization can produce highly conformal step-and-shoot treatment plans using only three to five apertures per beam direction. As compared with traditional optimization strategies, our studies demonstrate that direct aperture optimization can result in a significant reduction in both the number of beam segments and the number of monitor units. Direct aperture optimization therefore produces highly efficient treatment deliveries that maintain the full dosimetric benefits of IMRT. PMID- 12094971 TI - Deterministic calculations of photon spectra for clinical accelerator targets. AB - A method is proposed to compute photon energy spectra produced in clinical electron accelerator targets, based on the deterministic solution of the Boltzmann equation for coupled electron-photon transport in one-dimensional (1-D) slab geometry. It is shown that the deterministic method gives similar results as Monte Carlo calculations over the angular range of interest for therapy applications. Relative energy spectra computed by deterministic and 3-D Monte Carlo methods, respectively, are compared for several realistic target materials and different electron beams, and are found to give similar photon energy distributions and mean energies. The deterministic calculations typically require 1-2 mins of execution time on a Sun Workstation, compared to 2-36 h for the Monte Carlo runs. PMID- 12094972 TI - Monte Carlo calculations of dosimetry parameters of the urocor prostaseed 125I source. AB - This report presents the results of Monte Carlo calculations of the dosimetric parameters of the Urocor ProstaSeed 1251 seed source. This source contains five spherical silver markers, of 0.5 mm diameter, with 1251 deposited on the spheres through ion exchange. The silver spheres are then encapsulated within a 0.8 mm in diameter and 4.5 mm long cylindrical shell of titanium, as is common to this type of sources. Physical dimensions of the source are confirmed by measurement. Four (4) geometric models of the source, based on different assumptions on the locations of the silver spheres within the seed, were used in dose rate calculations. Monte Carlo photon transport simulation was used to calculate the dosimetric parameters of dose rate constant, radial dose function, and anisotropy function using these geometric models of the source. The Monte Carlo calculated dose rate constant of the ProstaSeed source was found equal to 0.925 cGy/Uh with approximate uncertainties of 5%. Radial dose function values and anisotropy function values were derived from the relative dose distribution around the source calculated by the Monte Carlo code. The calculated values of these dosimetric parameters agree with previously published thermoluminescence dosimeter-measured values for this source after consideration of measurement and calculation uncertainties. PMID- 12094973 TI - Benchmarking of the dose planning method (DPM) Monte Carlo code using electron beams from a racetrack microtron. AB - A comprehensive set of measurements and calculations has been conducted to investigate the accuracy of the Dose Planning Method (DPM) Monte Carlo code for dose calculations from 10 and 50 MeV scanned electron beams produced from a racetrack microtron. Central axis depth dose measurements and a series of profile scans at various depths were acquired in a water phantom using a Scanditronix type RK ion chamber. Source spatial distributions for the Monte Carlo calculations were reconstructed from in-air ion chamber measurements carried out across the two-dimensional beam profile at 100 cm downstream from the source. The in-air spatial distributions were found to have full width at half maximum of 4.7 and 1.3 cm, at 100 cm from the source, for the 10 and 50 MeV beams, respectively. Energy spectra for the 10 and 50 MeV beams were determined by simulating the components of the microtron treatment head using the code MCNP4B. DPM calculations are on average within +/- 2% agreement with measurement for all depth dose and profile comparisons conducted in this study. The accuracy of the DPM code illustrated in this work suggests that DPM may be used as a valuable tool for electron beam dose calculations. PMID- 12094975 TI - Considerations of anthropometric, tissue volume, and tissue mass scaling for improved patient specificity of skeletal S values. AB - It is generally acknowledged that reference man (70 kg in mass and 170 cm in height) does not adequately represent the stature and physical dimensions of many patients undergoing radionuclide therapy, and thus scaling of radionuclide S values is required for patient specificity. For electron and beta sources uniformly distributed within internal organs, the mean dose from self-irradiation is noted to scale inversely with organ mass, provided no escape of electron energy occurs at the organ boundaries. In the skeleton, this same scaling approach is further assumed to be correct for marrow dosimetry; nevertheless, difficulties in quantitative assessments of marrow mass in specific skeletal regions of the patient make this approach difficult to implement clinically. Instead, scaling of marrow dose is achieved using various anthropometric parameters that presumably scale in the same proportion. In this study, recently developed three-dimensional macrostructural transport models of the femoral head and humeral epiphysis in three individuals (51-year male, 82-year female, and 86 year female) are used to test the abilities of different anthropometric parameters (total body mass, body surface area, etc.) to properly scale radionuclide S values from reference man models. The radionuclides considered are 33P, 177Lu, 153Sm, 186Re, 89Sr, 166Ho, 32P, 188Re, and 90Y localized in either the active marrow or endosteal tissues of the bone trabeculae. S value scaling is additionally conducted in which the 51-year male subject is assigned as the reference individual; scaling parameters are then expanded to include tissue volumes and masses for both active marrow and skeletal spongiosa. The study concludes that, while no single anthropometric parameter emerges as a consistent scaler of reference man S values, lean body mass is indicated as an optimal scaler when the reference S values are based on 3D transport techniques. Furthermore, very exact patient-specific scaling of radionuclide S values can be achieved if measurements of spongiosa volume and marrow volume fraction (high resolution CT with image segmentation) are known in both the patient and the reference individual at skeletal sites for which dose estimates are sought. However, the study indicates that measurements of the spongiosa volume alone may be sufficient for reasonable patient-specific scaling of S values for the majority of radionuclides of interest in internal-emitter therapy. PMID- 12094974 TI - Theoretical analysis and experimental evaluation of a Csl(TI) based electronic portal imaging system. AB - This article discusses the design and analysis of a portal imaging system based on a thick transparent scintillator. A theoretical analysis using Monte Carlo simulation was performed to calculate the x-ray quantum detection efficiency (QDE), signal to noise ratio (SNR) and the zero frequency detective quantum efficiency [DQE(0)] of the system. A prototype electronic portal imaging device (EPID) was built, using a 12.7 mm thick, 20.32 cm diameter, Csl(Tl) scintillator, coupled to a liquid nitrogen cooled CCD TV camera. The system geometry of the prototype EPID was optimized to achieve high spatial resolution. The experimental evaluation of the prototype EPID involved the determination of contrast resolution, depth of focus, light scatter and mirror glare. Images of humanoid and contrast detail phantoms were acquired using the prototype EPID and were compared with those obtained using conventional and high contrast portal film and a commercial EPID. A theoretical analysis was also carried out for a proposed full field of view system using a large area, thinned CCD camera and a 12.7 mm thick CsI(TI) crystal. Results indicate that this proposed design could achieve DQE(0) levels up to 11%, due to its order of magnitude higher QDE compared to phosphor screen-metal plate based EPID designs, as well as significantly higher light collection compared to conventional TV camera based systems. PMID- 12094976 TI - A method for geometrical verification of dynamic intensity modulated radiotherapy using a scanning electronic portal imaging device. AB - In order to guarantee the safe delivery of dynamic intensity modulated radiotherapy (IMRT), verification of the leaf trajectories during the treatment is necessary. Our aim in this study is to develop a method for on-line verification of leaf trajectories using an electronic portal imaging device with scanning read-out, independent of the multileaf collimator. Examples of such scanning imagers are electronic portal imaging devices (EPIDs) based on liquid filled ionization chambers and those based on amorphous silicon. Portal images were acquired continuously with a liquid-filled ionization chamber EPID during the delivery, together with the signal of treatment progress that is generated by the accelerator. For each portal image, the prescribed leaf and diaphragm positions were computed from the dynamic prescription and the progress information. Motion distortion effects of the leaves are corrected based on the treatment progress that is recorded for each image row. The aperture formed by the prescribed leaves and diaphragms is used as the reference field edge, while the actual field edge is found using a maximum-gradient edge detector. The errors in leaf and diaphragm position are found from the deviations between the reference field edge and the detected field edge. Earlier measurements of the dynamic EPID response show that the accuracy of the detected field edge is better than 1 mm. To ensure that the verification is independent of inaccuracies in the acquired progress signal, the signal was checked with diode measurements beforehand. The method was tested on three different dynamic prescriptions. Using the described method, we correctly reproduced the distorted field edges. Verifying a single portal image took 0.1 s on an 866 MHz personal computer. Two flaws in the control system of our experimental dynamic multileaf collimator were correctly revealed with our method. First, the errors in leaf position increase with leaf speed, indicating a delay of approximately 0.8 s in the control system. Second, the accuracy of the leaves and diaphragms depends on the direction of motion. In conclusion, the described verification method is suitable for detailed verification of leaf trajectories during dynamic IMRT. PMID- 12094978 TI - Head insulation and heat loss in naked and clothed newborns using a thermal mannequin. AB - In newborns, large amounts of heat are lost from the head, due to its high skin surface area. Insulating the head (for example, with a hat or bonnet) can be a simple and effective method of reducing dry heat loss. In the present study, we evaluated the safety aspects of insulating the head of low-birth-weight naked or clothed newborns by using a heated mannequin that simulates a low-birth-weight newborn. Experimental conditions (comprising a nude and three clothed setups) were performed in a closed incubator at three different air temperatures (29 degrees C, 32 degrees C, and 34 degrees C) and with and without the head being covered with a bonnet in each case, i.e., 24 experimental conditions in all. The study shows that added clothing elements and insulating the head decreases the total heat loss of the mannequin as a whole. As regards the dry heat exchange from the head, wearing a bonnet decreases the local heat loss by an average of 18.9% in all clothed and thermal conditions. This phenomenon could be at the origin of brain overheating in heavily dressed newborns, when unrestricted heat loss is limited to the face only. Our results suggest that--apart from accidental hypothermia-in order to achieve thermal equilibrium of the body, it is preferable to leave the head unprotected and to increase the level of clothing insulation over the rest of the body. PMID- 12094977 TI - Comparisons of point and average organ dose within an anthropomorphic physical phantom and a computational model of the newborn patient. AB - Pediatric radiographic examinations yield medical benefits and/or diagnostic information that must be balanced against potential risk from patient radiation exposure. Consequently, clinical tools for measuring internal organ dose are needed for medical risk assessment. In this study, a physical phantom and Monte Carlo simulation model of the newborn patient were developed based upon their stylized mathematical expressions (ORNL and MIRD model series). The physical phantom was constructed using tissue equivalent substitutes for soft tissue, lung, and skeleton. Twenty metal-oxide-semiconductor field effect transistor (MOSFET) dosimeters were then inserted at three-dimensional positions representing the centroids of organs assigned in the ICRP's definition of the effective dose. MOSFET-derived point estimates of organ dose were shown to be in reasonable agreement with Monte Carlo estimates for representative newborn head, chest, and abdomen radiographic exams. Ratios of average organ dose assessed via MCNP simulations to the MOSFET-derived point doses (point-to-organ dose scaling factors, SF(POD)) are tabulated for subsequent use in clinical irradiations of the newborn phantom/MOSFET system. Values of SF(POD) indicate that MOSFET measurements of point dose for in-field exposures need to be adjusted only to within 10% to report volume-averaged organ dose. Larger adjustments to point doses are noted for organs out-of-field. For walled organs, point estimates of organ dose at the content centroid are shown to underestimate the average wall dose when the organ is within the primary field: SF(POD) of 1.19 for the stomach (AP chest exam), and SF(POD) of 1.15 for the urinary bladder (AP abdomen exam). PMID- 12094979 TI - Multitapered x-ray capillary optics for mammography. AB - X-ray mammography is currently the primary tool used for breast cancer detection. However, studies have shown that 5%-15% of breast cancers are not visualized mammographically. The long term goal of this project is to improve the x-ray mammographic imaging system using capillary optics. A post-patient capillary optic lens has the potential to increase spatial resolution and eliminate the detection of scattered x rays, thereby improving image contrast and the signal-to noise ratio (SNR). Several individual and two prototype multitapered optics were studied to determine the feasibility of a full-field multitapered optic. Scatter fraction, contrast, transmission, uniformity, and the modulation transfer function (MTF) were measured for a Mo target tube/computed radiography (CR) imaging system when this prototype was applied. The results were compared with standard grid and airgap techniques. The multitapered optic lens removed 85% of the scattered photons as compared to 66% and 39% for the air gap and grid methods, respectively. This resulted in an improvement of contrast by approximately 80% for the optics, 51% for the air gap, and 30% for grid methods. The single optic lens improved the limiting resolution (5% MTF level) of the CR detector by 78% due to magnification with very little focal spot blurring, while the multitapered prototype improved resolution significantly, but not as much as the single optic. These measurements have shown that it is feasible to create a multitapered optic lens that significantly improves system MTF and virtually eliminates scatter. With continued improvements in fabrication techniques, a full field multitapered lens will be feasible. PMID- 12094980 TI - Investigation of the use of MOSFET for clinical IMRT dosimetric verification. AB - (Received 22 October 2001; accepted for publication 26 March 2002; published 22 May 2002) With advanced conformal radiotherapy using intensity modulated beams, it is important to have radiation dose verification measurements prior to treatment. Metal oxide semiconductor field effect transistors (MOSFET) have the advantage of a faster and simpler reading procedure compared to thermoluminescent dosimeters (TLD), and with the commercial MOSFET system, multiple detectors can be used simultaneously. In addition, the small size of the detector could be advantageous, especially for point dose measurements in small homogeneous dose regions. To evaluate the feasibility of MOSFET for routine IMRT dosimetry, a comprehensive set of experiments has been conducted, to investigate the stability, linearity, energy, and angular dependence. For a period of two weeks, under a standard measurement setup, the measured dose standard deviation using the MOSFETs was +/- 0.015 Gy with the mean dose being 1.00 Gy. For a measured dose range of 0.3 Gy to 4.2 Gy, the MOSFETs present a linear response, with a linearity coefficient of 0.998. Under a 10 x 10 cm2 square field, the dose variations measured by the MOSFETs for every 10 degrees from 0 to 180 degrees is +/- 2.5%. The percent depth dose (PDD) measurements were used to verify the energy dependence. The measured PDD using the MOSFETs from 0.5 cm to 34 cm depth agreed to within +/- 3% when compared to that of the ionization chamber. For IMRT dose verification, two special phantoms were designed. One is a solid water slab with 81 possible MOSFET placement holes, and another is a cylindrical phantom with 48 placement holes. For each IMRT phantom verification, an ionization chamber and 3 to 5 MOSFETs were used to measure multiple point doses at different locations. Preliminary results show that the agreement between dose measured by MOSFET and that calculated by Corvus is within 5% error, while the agreement between ionization chamber measurement and the calculation is within 3% error. In conclusion, MOSFET detectors are suitable for routine IMRT dose verification. PMID- 12094981 TI - Effect of multileaf collimator leaf width on physical dose distributions in the treatment of CNS and head and neck neoplasms with intensity modulated radiation therapy. AB - The purpose of this work is to examine physical radiation dose differences between two multileaf collimator (MLC) leaf widths (5 and 10 mm) in the treatment of CNS and head and neck neoplasms with intensity modulated radiation therapy (IMRT). Three clinical patients with CNS tumors were planned with two different MLC leaf sizes, 5 and 10 mm, representing Varian-120 and Varian-80 Millennium multileaf collimators, respectively. Two sets of IMRT treatment plans were developed. The goal of the first set was radiation dose conformality in three dimensions. The goal for the second set was organ avoidance of a nearby critical structure while maintaining adequate coverage of the target volume. Treatment planning utilized the CadPlan/Helios system (Varian Medical Systems, Milpitas CA) for dynamic MLC treatment delivery. All beam parameters and optimization (cost function) parameters were identical for the 5 and 10 mm plans. For all cases the number of beams, gantry positions, and table positions were taken from clinically treated three-dimensional conformal radiotherapy plans. Conformality was measured by the ratio of the planning isodose volume to the target volume. Organ avoidance was measured by the volume of the critical structure receiving greater than 90% of the prescription dose (V(90)). For three patients with squamous cell carcinoma of the head and neck (T2-T4 N0-N2c M0) 5 and 10 mm leaf widths were compared for parotid preservation utilizing nine coplanar equally spaced beams delivering a simultaneous integrated boost. Because modest differences in physical dose to the parotid were detected, a NTCP model based upon the clinical parameters of Eisbruch et al. was then used for comparisons. The conformality improved in all three CNS cases for the 5 mm plans compared to the 10 mm plans. For the organ avoidance plans, V(90) also improved in two of the three cases when the 5 mm leaf width was utilized for IMRT treatment delivery. In the third case, both the 5 and 10 mm plans were able to spare the critical structure with none of the structure receiving more than 90% of the prescription dose, but in the moderate dose range, less dose was delivered to the critical structure with the 5 mm plan. For the head and neck cases both the 5 and 10 x 2.5 mm beamlets dMLC sliding window techniques spared the contralateral parotid gland while maintaining target volume coverage. The mean parotid dose was modestly lower with the smaller beamlet size (21.04 Gy v 22.36 Gy). The resulting average NTCP values were 13.72% for 10 mm dMLC and 8.24% for 5 mm dMLC. In conclusion, five mm leaf width results in an improvement in physical dose distribution over 10 mm leaf width that may be clinically relevant in some cases. These differences may be most pronounced for single fraction radiosurgery or in cases where the tolerance of the sensitive organ is less than or close to the target volume prescription. PMID- 12094982 TI - Experimental determination of the anisotropy function for the model 200 103Pd "light seed" and derivation of the anisotropy constant based upon the linear quadratic model. AB - Since the publication of the AAPM Task Group 43 report in 1995, Model 200 103Pd seed, which has been widely used in prostate seed implants and other brachytherapy procedures, has undergone some changes in its internal geometry resulting from the manufacturer's transition from lower specific activity reactor produced 103Pd ("heavy seeds") to higher specific activity accelerator-produced radioactive material ("light seeds"). Based on previously reported theoretical calculations and measurements, the dose rate constants and the radial dose functions of the two types of seeds are nearly the same and have already been reported. In this work, the anisotropy function of the "light seed" was experimentally measured and an averaging method for the determination of the anisotropy constant from distance-dependent values of anisotropy factors is presented based upon the continuous low dose rate irradiation linear quadratic model for cell killing. The anisotropy function of Model 200 103Pd "light seeds" was measured in a Solid Water phantom using 1 X 1 x 1 mm micro LiF TLD chips at radial distances of 1, 2, 3, 4, 5, and 6 cm and at angles from 0 to 90 degrees with respect to the longitudinal axis of the seeds. At a radial distance of 1 cm, the measured anisotropy function of the 103Pd "light seed" is considerably lower than that of the 103Pd "heavy seed" reported in the TG 43 report. Our measured values at all radial distances are in excellent agreement with the results of a Monte Carlo simulation reported by Weaver, except for points along and near the seed longitudinal axis. The anisotropy constant of the 103Pd "light seed" was calculated using the linear quadratic biological model for cell killing in 30 clinical implants. For the model 200 "light seed," it has a value of 0.865. However, our biological model calculations lead us to conclude that if the anisotropy factors of an interstitial brachytherapy seed vary significantly over radial distances anisotropy constant should not be used as an approximation for anisotropy characteristics of a brachytherapy seed. PMID- 12094983 TI - Dose optimization via index-dose gradient minimization. AB - This paper presents an iterative optimization algorithm based on gradient minimization of index dose, defined as the product of physical dose and a numerical index. Acting as a template the index distribution is designed to represent the dosimetry that meets the dose volume histogram-based optimization objectives. The treatment dosimetry is optimized when the uniformity of the index dose distribution is maximized. Prior to optimization the user can select all or only some of the beams to be intensity modulated. The remaining unmodulated beams can be either open or wedged photon beams, electron beams, or beams of previous treatments. The optimization result and treatment delivery efficiency can often be enhanced by including not only the IM photon beams but also all suitable fixed beams available on the linac in the treatment plan. In addition, the doses from previous treatments can also be considered in the optimization of current treatment. Five clinical examples with different complexities in optimization objective are presented. The effects of two nonoptimization variables, beam setup and initial beam weights, on the quality of the dose optimization are also presented. The results are analyzed in terms of isodose distribution, dose volume histograms, and a dose optimization quality factor. The optimization algorithm, implemented in our in-house TPS PLanUNC, has been used in clinical application since 1996. The primary advantages of our optimization algorithm include computational efficiency, intensity modulation selection choice, and performance reliability for a wide range of clinical beam setups and optimization objectives. PMID- 12094984 TI - Medical physicists need professional malpractice insurance. For the proposition. PMID- 12094986 TI - Portal dose image prediction for dosimetric treatment verification in radiotherapy. II. An algorithm for wedged beams. AB - A method is presented for calculation of a two-dimensional function, T(wedge)(x,y), describing the transmission of a wedged photon beam through a patient. This in an extension of the method that we have published for open (nonwedged) fields [Med. Phys. 25, 830-840 (1998)]. Transmission functions for open fields are being used in our clinic for prediction of portal dose images (PDI, i.e., a dose distribution behind the patient in a plane normal to the beam axis), which are compared with PDIs measured with an electronic portal imaging device (EPID). The calculations are based on the planning CT scan of the patient and on the irradiation geometry as determined in the treatment planning process. Input data for the developed algorithm for wedged beams are derived from (the already available) measured input data set for transmission prediction in open beams, which is extended with only a limited set of measurements in the wedged beam. The method has been tested for a PDI plane at 160 cm from the focus, in agreement with the applied focus-to-detector distance of our fluoroscopic EPIDs. For low and high energy photon beams (6 and 23 MV) good agreement (approximately 1%) has been found between calculated and measured transmissions for a slab and a thorax phantom. PMID- 12094987 TI - A simple method for bremsstrahlung spectra reconstruction from transmission measurements. AB - A new method for evaluation of bremsstrahlung spectra from transmission measurements has been developed. In this method some very well known facts relating to thick target bremsstrahlung spectra are a priori included in the calculation procedure. Some characteristics of the method are preliminarily illustrated on a 6 MV therapy linear accelerator. PMID- 12094988 TI - Experimental determination of the TG-43 dosimetric characteristics of EchoSeed model 6733 I25I brachytherapy source. AB - Recently an improved design of a 125I brachytherapy source has been introduced for interstitial seed implants, particularly for prostate seed implants. This design improves the in situ ultrasound visualization of the source compared to the conventional seed. In this project, the TG-43 recommended dosimetric characteristics of the new brachytherapy source have been experimentally determined in Solid Water phantom material. The measured dosimetric characteristics of the new source have been compared with data reported in the literature for other source designs. The measured dose rate constant, A, in Solid Water was multiplied by 1.05 to extract the dose rate constant in water. The dose rate constant of the new source in water was found to be 0.99 +/- 8% cGy h(-1) U( 1). The radial dose function was measured at distances between 0.5 and 10 cm using LiF TLDs in Solid Water phantom. The anisotropy function, F(r, theta), was measured at distances of 2, 3, 5, and 7 cm. PMID- 12094989 TI - Favorable noise uniformity properties of Fourier-based interpolation and reconstruction approaches in single-slice helical computed tomography. AB - Volumes reconstructed by standard methods from single-slice helical computed tomography (CT) data have been shown to have noise levels that are highly nonuniform relative to those in conventional CT. These noise nonuniformities can affect low-contrast object detectability and have also been identified as the cause of the zebra artifacts that plague maximum intensity projection (MIP) images of such volumes. While these spatially variant noise levels have their root in the peculiarities of the helical scan geometry, there is also a strong dependence on the interpolation and reconstruction algorithms employed. In this paper, we seek to develop image reconstruction strategies that eliminate or reduce, at its source, the nonuniformity of noise levels in helical CT relative to that in conventional CT. We pursue two approaches, independently and in concert. We argue, and verify, that Fourier-based longitudinal interpolation approaches lead to more uniform noise ratios than do the standard 360LI and 180LI approaches. We also demonstrate that a Fourier-based fan-to-parallel rebinning algorithm, used as an alternative to fanbeam filtered backprojection for slice reconstruction, also leads to more uniform noise ratios, even when making use of the 180LI and 360LI interpolation approaches. PMID- 12094990 TI - Incorporating multi-leaf collimator leaf sequencing into iterative IMRT optimization. AB - Intensity modulated radiation therapy (IMRT) treatment planning typically considers beam optimization and beam delivery as separate tasks. Following optimization, a multi-leaf collimator (MLC) or other beam delivery device is used to generate fluence patterns for patient treatment delivery. Due to limitations and characteristics of the MLC, the deliverable intensity distributions often differ from those produced by the optimizer, leading to differences between the delivered and the optimized doses. Objective function parameters are then adjusted empirically, and the plan is reoptimized to achieve a desired deliverable dose distribution. The resulting plan, though usually acceptable, may not be the best achievable. A method has been developed to incorporate the MLC restrictions into the optimization process. Our in-house IMRT system has been modified to include the calculation of the deliverable intensity into the optimizer. In this process, prior to dose calculation, the MLC leaf sequencer is used to convert intensities to dynamic MLC sequences, from which the deliverable intensities are then determined. All other optimization steps remain the same. To evaluate the effectiveness of deliverable-based optimization, 17 patient cases have been studied. Compared with standard optimization plus conversion to deliverable beams, deliverable-based optimization results show improved isodose coverage and a reduced dose to critical structures. Deliverable-based optimization results are close to the original nondeliverable optimization results, suggesting that IMRT can overcome the MLC limitations by adjusting individual beamlets. The use of deliverable-based optimization may reduce the need for empirical adjustment of objective function parameters and reoptimization of a plan to achieve desired results. PMID- 12094991 TI - Comparison of characteristics of photon and electron beams generated by Philips/Elekta and Varian linear accelerators. AB - Commissioning data of the common energy of 6 MV photon beams are analyzed for seven Philips/ Elekta linacs of five different models: SL 75/5, SL 15, SL 25, SLi Precise, and SLi. The observed beam quality %dd(10)x of 67.7% +/- 0.3 agrees with the manufacturer's specification of 67.5% +/- 2% but with a much smaller variation, suggesting the possibility of a more accurate beam quality guideline for acceptance commissioning of these linac models. Field size factors are less uniform, especially for the biggest field size of 40 x 40 cm2, but all agree within 1.1% between the five models. These 6 MV photon beams are also shown to be 0.4 MV harder than the Varian 2100C model. For commissioning electron beams generated by the Philips/Elekta models SL 15, SLi Precise and SLi, guidelines can be found from this work for the following parameters: actual field size at 100 SSD (up to +/- 0.6%) (different from manufacturer specified values at 95 SSD), and virtual source distance (up to +/- 0.7%). Significant differences are seen for R50 (up to 3.7 mm), sigma-theta-x, effective source distance, and cone size factor. However, except for R50 where Varian 2100C datasets are not available for comparison, the differences between the different Philips/Elekta models are comparable to those between the same Varian 2100C model linacs measured by us and Watts. PMID- 12094992 TI - A performance comparison of flat-panel imager-based MV and kV cone-beam CT. AB - The use of cone-beam computed tomography (CBCT) has been proposed for guiding the delivery of radiation therapy, and investigators have examined the use of both kilovoltage (kV) and megavoltage (MV) x-ray beams in the development of such CBCT systems. In this paper, the inherent contrast and signal-to-noise ratio (SNR) performance for a variety of existing and hypothetical detectors for CBCT are investigated analytically as a function of imaging dose and object size. Theoretical predictions are compared to the results of experimental investigations employing largearea flat-panel imagers (FPIs) at kV and MV energies. Measurements were performed on two different FPI-based CBCT systems: a bench-top prototype incorporating an FPI and kV x-ray source (100 kVp x rays), and a system incorporating an FPI mounted on the gantry of a medical linear accelerator (6 MV x rays). The SNR in volume reconstructions was measured as a function of dose and found to agree reasonably with theoretical predictions. These results confirm the theoretically predicted advantages of employing kV energy x rays in imaging soft-tissue structures found in the human body. While MV CBCT may provide a valuable means of correcting 3D setup errors and may offer an advantage in terms of simplicity of mechanical integration with a linear accelerator (e.g., implementation in place of a portal imager), kV CBCT offers significant performance advantages in terms of image contrast and SNR per unit dose for visualization of soft-tissue structures. The relatively poor SNR performance at MV energies is primarily a result of the low x-ray quantum efficiencies (approximately a few percent or less) that are currently achieved with FPIs at high energies. Furthermore, kV CBCT with an FPI offers the potential of combined volumetric and radiographic/fluoroscopic imaging using the same device. PMID- 12094993 TI - Direct conversion detectors: the effect of incomplete charge collection on detective quantum efficiency. AB - Direct conversion detectors offer the potential for very high resolution and high quantum efficiency for x-ray imaging, however, variations in signal can arise due to incomplete charge collection. A charge transport model was developed to describe the signal and noise resulting from incomplete charge collection. This signal to noise ratio (SNR) reduction was incorporated into the cascaded systems model for a simple x-ray detector. A new excess noise factor, A(c) (termed the collection noise factor) is introduced to describe the reduction in detective quantum efficiency (DQE). The DQE is proportional to the product of the quantum efficiency and the collection noise factor. If the trapping cross sections for electrons and holes are very different, and if the detector is biased improperly, the collection noise factor can drop to as low as 50%. In addition, the signal loss due to incomplete charge collection will reduce the DQE in the presence of added noise. Because of this, the DQE generally does not continue to improve with greater detector thickness. The collection noise factor and DQE are predicted for CdZnTe, PbI2, and Se. The optimization of detector thickness should be based not only on quantum efficiency but also on the charge collection statistics, which are influenced by bias field and polarity. PMID- 12094994 TI - A leaf sequencing algorithm to enlarge treatment field length in IMRT. AB - With MLC-based IMRT, the maximum usable field size is often smaller than the maximum field size for conventional treatments. This is due to the constraints of the overtravel distances of MLC leaves and/or jaws. Using a new leaf sequencing algorithm, the usable IMRT field length (perpendicular to the MLC motion) can be mostly made equal to the full length of the MLC field without violating the upper jaw overtravel limit. For any given intensity pattern, a criterion was proposed to assess whether an intensity pattern can be delivered without violation of the jaw position constraints. If the criterion is met, the new algorithm will consider the jaw position constraints during the segmentation for the step and shoot delivery method. The strategy employed by the algorithm is to connect the intensity elements outside the jaw overtravel limits with those inside the jaw overtravel limits. Several methods were used to establish these connections during segmentation by modifying a previously published algorithm (areal algorithm), including changing the intensity level, alternating the leaf sequencing direction, or limiting the segment field size. The algorithm was tested with 1000 random intensity patterns with dimensions of 21 x 27 cm2, 800 intensity patterns with higher intensity outside the jaw overtravel limit, and three different types of clinical treatment plans that were undeliverable using a segmentation method from a commercial treatment planning system. The new algorithm achieved a success rate of 100% with these test patterns. For the 1,000 random patterns, the new algorithm yields a similar average number of segments of 36.9 +/- 2.9 in comparison to 36.6 +/- 1.3 when using the areal algorithm. For the 800 patterns with higher intensities outside the jaw overtravel limits, the new algorithm results in an increase of 25% in the average number of segments compared to the areal algorithm. However, the areal algorithm fails to create deliverable segments for 90% of these patterns. Using a single isocenter, the new algorithm provides a solution to extend the usable IMRT field length from 21 to 27 cm for IMRT on a commercial linear accelerator using the step and shoot delivery method. PMID- 12094995 TI - Quantification of the effect of kvp on edge-enhancement index in phase-contrast radiography. AB - This study was performed to measure the dependence of edge-enhancement in polychromatic phase-contrast radiography on x-ray tube operating voltage. Measurements of edge enhancement were made at tube voltages from 40 to 86 kVp using a tungsten anode x-ray tube with a nominal focal spot size of 100 micrometers. A relatively weak attenuating, sharp edge consisting of a thin lucite sheet (3 mm) in air was imaged utilizing phase-contrast radiography (PC R). PC-R images were acquired at different radiographic techniques in which x-ray tube voltage was varied from 40 to 86 kVp. The image receptor was a single emulsion x-ray mammography cassette. Optical density profiles across the edge of the object were obtained using a film digitizer and edge-enhancement indices were calculated. Increasing kVp resulted in a gradual decrease of the edge-enhancement index. Even at the highest kVp (86), however, important edge-enhancement effects were evident. While there is some degradation in the edge-enhancement effect of phase-contrast radiography at higher kVps, the decrease from 40 to 86 kVp is relatively small (11%). Our results suggest that further investigation into the role of phase-contrast imaging at higher kVp values for the purpose of patient dose reduction while still realizing the advantage of phase-contrast effects for improved soft-tissue detectability is warranted. PMID- 12094996 TI - Optimizing long-term response to methadone maintenance treatment: a 152-week follow-up using higher-dose methadone. AB - We report the clinical course over 152 weeks of 245 patients in methadone maintenance treatment: 144 high dose (HD) patients (> or = 100 mg/d, mean 211 mg/d), and 101 control (C) patients (< 100 mg/d, mean 65 mg/d). After 152 weeks the mean methadone doses were 284.9 mg/d (range 13-1100 mg/d) and 94.0 mg/d (range 10-500 mg/d), respectively. Overall retention in treatment was 59% over the 152 weeks, with the HD group having significantly better retention (61.1% vs. 46.3%) and lower rates of positive urine toxicologies (16.0% vs. 36.6%). Mortality was statistically the same for the HD group (2/144, 1.4%) and the C group (2/101, 1.9%) over the 152-week period. We conclude that doses of methadone exceeding 100 mg/d are safe and effective in long-term maintenance treatment. We attribute the favorable outcomes we report to a model of treatment that emphasizes medication management in the treatment of opioid addiction. PMID- 12094998 TI - The impact of treatment on criminalized substance addicts. AB - The objective of this research is to evaluate the impact of a treatment program specialized with criminalized addicts. In all, 119 criminalized addicts were admitted to the experimental procedure (specialized treatment) while 30 were admitted to a comparison group (general treatment). Both groups were contacted at five, eight, and eleven month intervals following the onset of the treatment process. Results showed that both groups improved. Those admitted to the unspecialized treatment improved earlier (in the first five months) than clients referred to the specialized treatment facility, however, after eight months, the improvement was similar for both groups. PMID- 12094997 TI - The use of levo-alpha-acetylmethadol (LAAM) in methadone patients who have not achieved heroin abstinence. AB - Levo-alpha-acetylmethadol (LAAM) pharmacotherapy was offered to twelve patients who continued illicit opioid abuse after > or = eleven months in methadone maintenance treatment. After 6-8 weeks on LAAM, plasma concentrations of the norLAAM metabolite varied significantly by LAAM dosing day, plasma adrenocorticotropin (ACTH) concentrations were significantly increased compared to methadone, and two of the seven subjects remaining in LAAM treatment were free of illicit opioids and nonprescribed methadone. After one year, one of five remaining subjects was using illicit opioids, and three were using non-prescribed methadone. While subject acceptance of LAAM was high, subjects were not in a "steady-state," with evidence of ongoing illicit opioid abuse. PMID- 12094999 TI - Comorbid psychiatric and alcohol abuse/dependence disorders: psychosocial stress, abuse, and personal history factors of those in treatment. AB - Factors related to comorbid versus only substance disorders are essential to understanding and treating these complex problems. Medical records of sixty-nine inpatients at a private rehabilitation hospital in Southern California were reviewed to determine the associations between personal history factors and (1) comorbid psychiatric and substance abuse disorders and (2) participant's self assessed progress in treatment. Results revealed significant differences between dual diagnosis patients (alcohol abuse/dependence and an affective disorder) and alcohol abuse/dependence only in regard to gender, previous diagnosis, length of illness, suicide attempts, psychotropic medication history, maternal emotional, physical and sexual abuse, paternal abuse, legal difficulties, and psychosocial stressors. No significant differences between substance abusing patients and dually diagnosed patients were found in terms of self-assessment of progress in treatment. Significant correlations were found between self-assessed progress in treatment and major depression (versus bipolar disorder), use of psychotropic medication, and less abuse from mother or primary caretaker. Identification of these personal history factors may be useful in developing and implementing treatment plans. PMID- 12095000 TI - Correlates of HIV transmission risk factors and considerations for interventions in homeless, chemically addicted and mentally ill patients. AB - A study was conducted to ascertain correlates of HIV high risk behaviors and attitudes toward HIV. A questionnaire was administered to 103 men living in a modified therapeutic community (TC) for homeless, chemically addicted and mentally ill men. The psychiatric diagnoses of the sample population included psychotic disorders (48%), depressive disorders (36%), and bipolar disorders (16%). Forty-two percent reported that their primary substance of abuse was cocaine and another 40% named alcohol as the substance to which they were most addicted. Two logistic regression analyses were conducted, one with needle sharing as the outcome measure and one with endorsement of the need for lifestyle changes to reduce risk of HIV transmission. Cocaine users were 3.4 times more likely to have shared needles than the rest of the sample. Patients who had a history of sexually transmitted diseases (STDs) were 17 times more likely to endorse the need for lifestyle changes. The level of HIV transmission knowledge was unrelated to HIV risk behaviors or attitudes. PMID- 12095001 TI - Schizoid phenomena in substance abusers. AB - It is hypothesized that the spectrum of schizoid disorders, schizoid phenomena, and the underlying psychodynamics can often be found in the gamut of addictions and stand in the way of recovery. Features of schizoidness, the varieties of schizoid presentations, the etiology and pathogenesis of drug/alcohol abuse in the schizoid, and readily clinically apparent psychodynamic features are discussed. Schizoid phenomena can be dealt with effectively with an informed psychotherapy. PMID- 12095002 TI - Physician behavior towards male and female problem drinkers: a controlled study using simulated patients. AB - BACKGROUND: Evidence suggests that physicians are less likely to identify alcohol problems in females than in males. PURPOSE: To compare the performance of family medicine residents with male and female simulated patients (SPs) posing as problem drinkers. METHODS: Fifty-six family medicine residents completed a baseline survey on knowledge and attitudes towards problem drinkers. Each resident was then visited by one male and female unannounced SP. The male and female roles were similar with respect to presenting complaint (in somnia or hypertension), age, social class, and drinking history. RESULTS: Residents expressed slightly more positive attitudes towards female than male patients (3.32 vs. 3.09, p < .001). Residents scored higher with undetected male than with undetected female SPs on the assessment checklist (5.1 vs. 3.2, p < .045), the management checklist (4.4 vs. 3.2, p = .032), and an interpersonal rating scale (the Alcohol Skills Rating Form; 5.5 vs. 4.7, p = .023). CONCLUSION: Educational programs should focus on improving physicians' clinical skills in the identification and treatment of alcohol problems in women. PMID- 12095003 TI - Transforming a health care system by transforming professional practice: a conversation with Gail Wolf, RN, DNS, FAAN. PMID- 12095004 TI - Nursing work force in practice and education: what can we learn from the current crisis? AB - A major factor in the growing crisis in health care is the serious shortage of nurses in practice and education. The current work force shortage is different and more critical than previous cyclical deficits. Because it is projected to be of unprecedented severity and to endure long into the future, it demands significant attention and innovation. Nurses in clinical practice and in faculty roles are growing older, and the nursing work force in general is becoming relatively less educated. A strong nursing work force of the future will require new approaches to recruitment, preparation, and retention of nurses, interdisciplinary partnerships, and infusion of support from a variety of sources. PMID- 12095005 TI - Developing a shared leadership model at the unit level. AB - As the nursing profession again faces a serious shortage, many are seeking ways to attract and retain talented people in the nursing profession. Creating both empowering and healthy work environments is a way to achieve this goal. Shared leadership is a nursing management model that supports staff nurses in extending their influence about decisions that affect their practice, work environment, professional development, and self-fulfillment. It is a way to strengthen continuous learning and enhance relationships, which are the foundation upon which nurses can develop a new type of relationship with management and with each other. PMID- 12095006 TI - Root cause analysis in perinatal care: health care professionals creating safer health care systems. AB - Medical error is a leading cause of death in the United States. The root cause analysis (RCA) process provides the framework for analysis of sentinel events and other medical errors. A thorough understanding of this process is a key component in promoting safety within the health care setting. Risk reduction strategies are presented to make this process meaningful and efficient while promoting the ultimate goal of safer health care systems. PMID- 12095007 TI - Managing women's and children's services: contemporary models as a template for the future. AB - Historically hospitals have struggled with organizational design and management of services within a single institution. The traditional design has been a bureaucracy with a hierarchical management structure. As individual hospitals develop new business relationships to form health systems, there is a need for innovative solutions that provide the flexibility and responsiveness necessary for successful health care management. One traditional business model, the SBU, or strategic business unit, may serve as a template for the development of product or service lines. The SBU has application for selected portions of a health care organization. The SBU will be discussed as it specifically relates to women's and children's health care services. PMID- 12095008 TI - Designing and delivering neonatal care in single rooms. AB - This article will outline the phases of the design process and describe the outcomes as related to a specific neonatal intensive care design project. The single-room concept allows for increased privacy and confidentiality, as well as decreased stress of family, staff, and especially patients. The improvement in family space facilitates improved opportunities for encouraging parent and sibling participation in care. Specific developmental and technological improvements have also been gained and are outlined throughout the article. PMID- 12095009 TI - Nursing work force of the future: the administrative perspective. AB - The nursing work force shortage is having a major impact on an industry that is already reeling from major assaults driven by inadequate payment and reimbursement. This article evaluates the current work force shortage as it relates to the competitive marketplace, the data regarding the shortages, the work environment, and the intergenerational issues that sabotage teams. There are recommendations for action in the areas of recruitment and retention, training and education, leadership and management, and improving the work environment. PMID- 12095010 TI - Cholinesterase estimations revisited: the clinical relevance. PMID- 12095011 TI - Central neuraxial blocks and anticoagulation: a review of current trends. AB - Patients receiving anticoagulants offer a challenge to anaesthesiologists. The issue of spinal haematoma following central neuraxial block in such patients is a contentious issue. Although rare, with an estimated incidence of < 1:150,000 for epidural blocks and 1:220,000 for spinal anaesthetics in patients with normal coagulation status, this is an emergency situation with a potentially grave prognosis. The review presents cases of spinal haematomata that have occurred in the last 5 years, both spontaneously and after central neuraxial blockade. Of the 60 cases reported in the literature, 33% occurred following central neuraxial block and, of these, 55% were associated with concomitant use of anticoagulants. The pharmacology of the newer and older anticoagulants is also described. The variety of risk factors and diverse recommendations that have been described in these patients are reviewed. PMID- 12095012 TI - Effects of normovolaemic haemodilution on middle cerebral artery blood flow velocity and oxygen delivery. AB - BACKGROUND AND OBJECTIVE: Assessment of the effects of normovolaemic haemodilution on middle cerebral artery blood flow velocity with transcranial Doppler ultrasonography, intracranial pressure, cerebral perfusion pressure, arterial oxygen content and cerebral oxygen delivery. METHODS: Normovolaemic haemodilution was induced in rabbits under general anaesthesia, and the haematocrit was allowed to decrease to 30% in Group 1 (n = 6) and to 20% in Group 2 (n = 6). Peak systolic and diastolic velocities, mean blood flow velocity, and pulsatility and resistance indices of the middle cerebral artery were measured by transcranial Doppler ultrasonography. Changes in intracranial pressure, cerebral perfusion pressure, arterial oxygen content and cerebral oxygen delivery were also assessed. RESULTS: In Group 2, middle cerebral artery blood flow velocity increased from 0.4 +/- 0.01 to 0.51 +/- 0.02 m s(-1) after the induction of normovolaemic haemodilution (P = 0.04), while arterial oxygen content decreased from 16.2 +/- 0.1 to 8.5 +/- 0.1 mLdL(-1) (P = 0.002). The decrease in cerebral oxygen delivery from 6.5 +/- 0.2 to 4.3 +/- 0.2 was also significant (P = 0.02). However, no associated changes in intracranial pressure and cerebral perfusion pressure could be demonstrated. CONCLUSIONS: Normovolaemic haemodilution resulted in an increase in the mean blood flow velocity of the middle cerebral artery. However, this increase did not compensate for the consequences of the altered oxygen delivery to the brain when the haematocrit was reduced to 20%. PMID- 12095013 TI - Cardiac output assessed by arterial thermodilution during exsanguination and fluid resuscitation: experimental validation against a reference technique. AB - BACKGROUND AND OBJECTIVE: The arterial thermodilution technique offers the ability to measure cardiac output using only central venous and arterial catheters. However, the technique has been reported to overestimate cardiac output because of a higher loss of cold indicator due to the increased distance between the sites of injection and measurement. In this study, the two techniques were compared with respect to conditions of low cardiac output in which a longer passage time may further increase loss of indicator. METHODS: Seventeen anaesthetized dogs were studied during hypovolaemic shock and fluid resuscitation. Cardiac output measurements were carried out simultaneously by arterial and pulmonary artery thermodilution techniques. RESULTS: One-hundred-and two measurements were performed. The mean cardiac output was 2.28 +/- 1.4Lmin(-1) by the pulmonary arterial technique and 2.29 +/- 1.56Lmin(-1) by the arterial thermodilution technique. The correlation coefficient between the two measurements was 0.95, the precision -0.04 +/- 0.41 Lmin(-1) and the limits of agreement from -0.86 to 0.78Lmin(-1). The agreement was also consistent at low cardiac outputs. CONCLUSIONS: The arterial thermodilution technique may serve as a less invasive cardiac output monitor in conditions of severe bleeding and shock. PMID- 12095014 TI - Fluoride ion toxicity in rabbit kidney thick ascending limb cells. AB - BACKGROUND AND OBJECTIVE: Some halogenated agents, especially methoxyflurane, because of a higher level of fluoride production, induce a renal concentrating defect that could be related to an ascending limb impairment. We investigated the mechanisms of fluoride toxicity on an immortalized cell line. METHODS: Cells were cultured for 2, 6 or 24 h in the presence of fluoride. Toxicity evaluation was based on: cell numbers, protein content, leucine-incorporation, lactate dehydrogenase (LDH) and N-acetyl-beta-glucosaminidase (NAG) releases, Na-K-ATPase and Na-K-2Cl activities, electron microscope studies. Infrared analysis and fluoride microdetermination allowed crystal components. RESULTS: At 5 mmol after 24 h, fluoride decreased cell numbers (-14%, *P < 0.05), protein content (-16%*), leucine incorporation (-54%*), Na-K-2Cl activity (-84%*), increased LDH (+145%*) and NAG release (+190%*). Na-K-ATPase was more sensitive and impaired from 1 mmol for 24h and after 2 h at 5 mmol. Crystal formation in mitochondria occurred after 6 h at 5 mmol. Infra-red analysis and fluoride microdetermination established that crystals contained sodium, phosphate and fluoride. CONCLUSIONS: The results suggest that the Na-K-ATPase pump is a major target for fluoride toxicity in Henle's loop. PMID- 12095015 TI - Comparison of the effects of remifentanil or fentanyl on anaesthetic induction characteristics of propofol, thiopental or etomidate. AB - BACKGROUND AND OBJECTIVE: This prospective, randomized, double-blinded study was designed to compare the effects of remifentanil or fentanyl on anaesthetic induction characteristics of propofol, thiopental or etomidate. METHODS: Seventy two patients were enrolled in six groups of 12 individuals each. In three groups, fentanyl was given as a bolus dose of 1.5 microg kg(-1), whereas the others received a remifentanil infusion at 0.5 microg kg(-1) min(-1). Five minutes later, propofol, thiopental or etomidate were titrated to a state of unresponsiveness. Assessment included the amounts of drug necessary for induction, haemodynamics and the times to apnoea, loss of eyelash reflex, and the release of a water-filled syringe held in the patient's hand. RESULTS: Induction times to loss of the eyelash reflex were significantly shorter in the remifentanil than in the fentanyl groups: with propofol 50.7 +/- 13.6s (mean +/- SD) versus 74.9 +/- 27.0s (P < 0.01), with thiopental 42.9 +/- 16.8s versus 77.2 +/- 27.8s (P < 0.01) and with etomidate 54.7 +/- 17.6s versus 72.3 +/- 24.0s (P < 0.05). The times to respiratory arrest or for the syringe to fall were significantly shorter with remifentanil than with fentanyl for propofol and for thiopental, but not for etomidate. In terms of dosages per kg body weight necessary to achieve unresponsiveness, less propofol (-29%, P < 0.05), thiopental (-25%, P < 0.05) or etomidate (-32%, P < 0.01) was necessary with remifentanil than with fentanyl. Haemodynamic responses to tracheal intubation were controlled more effectively with remifentanil. However, within the remifentanil groups, mean arterial pressure significantly decreased during induction: -26% with propofol, 181% with thiopental and -14% with etomidate (all P < 0.01). CONCLUSIONS: During anaesthetic induction, a remifentanil infusion of 0.5 microg kg(-1) min(-1) over 5 min is a suitable alternative to a 1.5 microg kg(-1) bolus dose of fentanyl: induction times are shorter with reduced amounts of propofol, thiopental or etomidate. PMID- 12095016 TI - Delivery performance of commercial target-controlled infusion devices with Diprifusor module. AB - BACKGROUND AND OBJECTIVE: We measured the physical performance error of commercial syringe pumps incorporating the Diprifusor module to deliver propofol by target-controlled infusion in a laboratory experiment. METHODS: Three infusion devices (Graseby 3500; Fresenius Vial Master TCI; Alaris TIVA-TCI) were studied when infusing propofol at six different target concentrations (1, 1.5, 2, 3, 5, 8 microg mL(-1)) over 10 min. This procedure was repeated five times to average each target. Precision measurements were based on the volume delivered and a mean performance error was calculated. RESULTS: The mean performance error bandwidth for all pumps was from -2.07 to 4.22% with a 95% confidence interval always <4.5%. There was a higher variability at lower target concentrations associated with lower infusion rates. The Alaris TIVA-TCI pump showed the largest overall bias whereas the Fresenius Vial Master TCI pump had the smallest divergence and the lowest variation across all target concentrations. The Graseby 3500 pump had the smallest range in mean performance error with an acceptable variation (95% CI <2%). CONCLUSIONS: Despite the differences between the pumps, which may be related to the synchronization of the hardware components, the absolute inaccuracies in the physical performance were low and presumably negligible from a clinical point of view. PMID- 12095017 TI - Succinylcholine improves the time of performance of a tracheal intubation in prehospital critical care medicine. AB - BACKGROUND AND OBJECTIVE: The study aimed to evaluate and improve airway management in the prehospital setting, i.e. physicians working on board ambulances. A quality control programme focusing on anaesthesia was instituted to improve the time taken to perform endotracheal intubation. METHODS: All consecutive patients requiring tracheal intubation were prospectively analysed before (first period) and after the training programme focusing on anaesthetic protocols for tracheal intubation (second period). The number of attempts at laryngoscopy, the time taken to achieve tracheal intubation, the difficulties encountered and the related complications of the anaesthetic technique were recorded. At the end of the first period, the results were reported to the whole staff of the unit and the anaesthesia protocols were then modified by introducing succinylcholine into the induction sequence, as part of a training programme. RESULTS: Two-hundred-and-eighty patients were evaluated (97 in the first period, 183 in the second). All patients were successfully intubated in both periods. The percentage of difficult intubations (as assessed by the physician) was lower in the second period (20 versus 35%, respectively; P < 0.01). The success rate at the first attempt was significantly higher (74% [68-80] 95% CI versus 55% [45-65] 95% CI, P < 0.01) and the duration of intubation was significantly shorter in the second period than in the first (1.4 +/- 3.2 vs. 4.1 +/- 6.7 min, respectively; P < 0.001). The incidence of complications (hypoxaemia, laryngospasm, bronchospasm) was lower in the second period (15 versus 31%, P < 0.01). CONCLUSIONS: The time to perform tracheal intubation can be improved by the introduction of succinylcholine into the prehospital anaesthetic protocol. Rapid sequence induction should be taught as a way of improving tracheal intubation in the prehospital setting. PMID- 12095018 TI - Intravenous single-dose tramadol versus meperidine for pain relief in renal colic. AB - BACKGROUND AND OBJECTIVE: Comparison of the effectiveness of tramadol with meperidine given intravenously to emergency patients with suspected renal colic. METHODS: A double-blind, randomized clinical trial was performed in the Emergency Department of a tertiary-care university hospital. Consecutive patients with suspected renal colic (n = 47) were randomized to receive intravenously an initial dose of tramadol 50 mg (n = 23) or meperidine 50 mg (n = 24). After 30 min, additional doses of meperidine 50 mg were given intravenously as a rescue medication in an open fashion. Pain relief was assessed using a 10 cm visual analogue scale, the primary outcomes being pain relief at 15 and 30 min after the analgesics. Secondary outcomes were the frequency of rescue meperidine use and the development of side-effects. RESULTS: Visual analogue scale pain scores after 15 and 30 min decreased in both tramadol and meperidine groups (P < 0.05). However, pain relief was better in the meperidine group at the 15 and 30 min evaluations (P < 0.05). Only 11 patients (48%), initially receiving meperidine, needed more meperidine compared with 16 patients (67%) initially receiving tramadol. Both drugs were well tolerated with no adverse effects occurring in either group. CONCLUSIONS: Meperidine 50 mg was superior to tramadol 50 mg for acute pain relief in patients with suspected renal colic when given intravenously. Because many patients in both groups received supplemental meperidine and the response to tramadol alone cannot be predicted, clinicians may want to choose higher doses of meperidine alone or other alternative combinations. PMID- 12095019 TI - Conditions for insertion of the laryngeal mask airway: comparisons between sevoflurane and propofol using fentanyl as a co-induction agent. A pilot study. AB - BACKGROUND AND OBJECTIVE: To compare the conditions for insertion of the laryngeal mask airway using sevoflurane or propofol plus fentanyl. We evaluated the haemodynamic changes and cost of induction of anaesthesia in both groups. METHODS: Sixty patients were equally and randomly divided into two groups. Both groups received fentanyl 1 microg kg(-1). Patients in the sevoflurane group were induced with 8% sevoflurane and those in the propofol group with propofol 2.5 mg kg(-1). Conditions for insertion were graded on a three-point scale using six variables. Overall, conditions were assessed as excellent, satisfactory or poor based on the total score in each group. Systolic and diastolic arterial pressure and heart rate were recorded for 6 min after mask insertion. The financial cost of induction in both groups was calculated. RESULTS: The mean (+/- SD) time taken from induction to successful laryngeal mask insertion was significantly shorter with propofol (68.70 +/- 22.60 s) compared with sevoflurane (149.83 +/- 55.25 s). Excellent or satisfactory conditions were observed in 30 (100%) patients in the propofol group and in 29 (96.66%) in the sevoflurane group. Systolic and diastolic arterial pressures were significantly lower in the propofol group. The cost of sevoflurane used was 3.95 euros +/- 1.48 (Rs 216.23 +/- 64.66) (P < 0.05) compared with that of propofol, which was 3.23 euros +/- 0.65 (Rs 141.00 +/- 28.20). CONCLUSIONS: Although there was a faster induction with propofol fentanyl, conditions for insertion of the laryngeal mask airway were similar in both groups. Haemodynamic stability was better with sevoflurane-fentanyl. The propofol-fentanyl combination was more cost-effective. PMID- 12095020 TI - Aetiology of convulsions due to stellate ganglion block: a review and report of two cases. AB - Stellate ganglion block is a selective sympathetic block that affects the ipsilateral head, neck, upper extremity and upper part of the thorax. Convulsions are a recognized complication of intra-arterial injection during stellate ganglion block. As central nervous system toxicity depends ultimately on the concentration of the local anaesthetics presented to the brain, the likely causative factors are discussed as well as the types of toxic symptoms and their onset times. The paper considers the aetiological factors of such convulsions resulting from stellate ganglion block in two patients. PMID- 12095021 TI - Monitoring in neuroanaesthesia: update of clinical usefulness. PMID- 12095022 TI - Gum elastic bougie, capnography and apnoeic oxygenation. PMID- 12095023 TI - Systemic heparinization during postoperative pulmonary embolism induces fatal complications. PMID- 12095024 TI - Intravenous regional blocks with guanethidine and prilocaine combined with physiotherapy: two children with complex regional pain syndrome, type 1. PMID- 12095025 TI - Health risks and psychosocial outcomes of early childbearing: a review of the literature. AB - This article presents a review of the literature on the obstetric, medical, and neonatal health risks as well as psychosocial outcomes associated with early childbearing. An important concern in adolescent pregnancies is the increased risk for infant morbidity and mortality as a result of higher incidence of preterm births and low-birthweight infants. Implications for prenatal and postpartum nursing care are discussed. Promising new models to improve care of young mothers and their children, such as home visitation by public health nurses, are presented and features of successful intervention programs outlined. PMID- 12095026 TI - The theater of birth: scenes from women's scripts. AB - An analogy between theater and birth is drawn from analyses of women's birth stories to describe birth from a fresh perspective. Birth and theater are compared using the theatrical production elements: setting, casting, props, set, behind the scenes, script, and roles. Selected examples from women's birth stories highlight each element. Nurses' roles are significant during labor and birth, but nurses' abilities to fulfill these roles are threatened. This analogy promotes rethinking of nursing actions in the theater of birth. Implications for clinical practice are provided, including altering the birth environment, offering choices, and maintaining the woman's role as star. PMID- 12095027 TI - Eating disorders, fertility, and pregnancy: relationships and complications. AB - Women are becoming heavier with each generation although the ideal female image emphasizes slimness. This focus results in the development of eating disorders in a significant number of women. The most common eating disorders are anorexia nervosa and bulimia nervosa. Eating disorder behaviors during pregnancy are associated with complications such as preterm delivery, low birthweight, intrauterine growth restriction, Caesarean birth, and low Apgar scores. Increasing the understanding of eating disorders assists health care professionals to accurately assess and intervene to improve a woman's nutritional status, monitor eating behaviors that may negatively affect a woman's health and fertility, and promote positive outcomes during pregnancy. PMID- 12095028 TI - Struggling to mother: a consequence of inhibitive nursing interactions in the neonatal nursery. AB - This paper reports on one aspect of a grounded theory study of women's experiences of mothering in the nursery. Over 60 hours of interview data with 28 Australian women were analyzed. The analysis revealed that mothering in the nursery was a three-way interaction. "Struggling to mother" was the label given to the major category that represented how women responded to unsatisfactory nursing encounters. The findings illuminate how inhibitive nursing interactions relegate women to the periphery of care and engender an array of negative emotional responses that leave women feeling inconsequential to the welfare of their infant. Ultimately this has implications for how women take up their role as mothers in the nursery and for their sense of confidence, competence, and connection with their infant. PMID- 12095029 TI - Mothers' strategies to meet their needs in the newborn intensive care nursery. AB - In this prospective, critical ethnographic study using a total of 448 hours of participant observations incorporating in-depth interviews, the author explored with 12 mothers of premature babies their descriptions and interpretations of their own needs in a tertiary-level newborn nursery (NICU) and their actions to address these needs. In the NICU the mothers demonstrated a repertoire of actions to vigilantly "watch over" their premature babies, which they extended to address their own needs as well. The mothers employed these actions in such a way as to minimize the potential for diverting resources from their babies and to minimize conflict with caregivers. PMID- 12095030 TI - AJT 2001: the genesis. PMID- 12095032 TI - Clinical liver transplantation. PMID- 12095031 TI - Transplantation in the HIV+ patient. AB - The historical exclusion from transplantation of HIV-infected people was based on the logical premise that immunosuppression required for organ transplantation would exacerbate an immunocompromised state. However, the prognosis for people with HIV infection has dramatically improved with the clinical use of highly active antiretroviral (ARV) therapy (HAART). Clinical trials of ARV agents have demonstrated significant virologic, immunologic and survival benefits associated with the use of protease inhibitor (PI) or non-nucleoside reverse transcriptase inhibitor (NNRTI) containing regimens, when combined with two nucleoside analogues. The incidence of opportunistic infections and hospitalizations has decreased with the use of HAART. In combination with historical data suggesting that a subpopulation of HIV+ transplant recipients tolerate immunosuppression and have allograft survival comparable to that of HIV- transplant recipients, these results indicate that the medical community should readdress HIV infection as a contraindication to transplantation. PMID- 12095033 TI - T-cell-mediated rejection of vascularized xenografts in the absence of induced anti-donor antibody response. AB - T cells are considered to play a major indirect role in the pathogenesis of xenograft vascular rejection, by promoting the induction of anti-donor antibodies that trigger complement- and antibody-dependent cell cytotoxicity. However, how vigorous the T cell xenoresponse is in vivo, and whether, besides their helper function, T cells are capable of directly affecting the graft is still unclear. We have previously shown that cyclosporine A (CsA) withdrawal in accommodated cardiac xenograft recipient allows for a rapid and dense T-cell infiltration, concomitant to an acute graft rejection. In this paper we further characterize the role of T cells in this rejection process and we demonstrate that adoptive transfer of CD4+ T cells in irradiated recipients of long-term cardiac xenografts is sufficient to trigger acute rejection, in the absence of any detectable induced anti-hamster antibody response. Therefore, our data suggest that unusually strong T-cell response will be another major barrier to xenotransplantation, even if antibody-mediated vascular rejection is controlled. PMID- 12095034 TI - Initial inhibition of tissue factor signalling reduces chronic vascular changes in isogenic rat aortic transplants. AB - Vascular changes are considered the major histopathological indicator of chronic allograft dysfunction. These changes are characterized by intimal thickening caused by accumulation of primarily smooth muscle cells. Contributing factors may be of both immunological and nonimmunological origin. Cold ischemia has been shown to trigger intimal proliferation in the absence of alloantigen in an isogenic rat aortic transplant model. We have used this model to investigate the impact of inhibition of tissue factor (TF) signalling on the progression of intimal thickening. Group 1 was treated with recombinant FVIIa inhibited in its active site (rFVIIai), and group 2 served as untreated controls. At 8 weeks the intimal area was measured with image analysis. Medial areas and the proportion of medial necrosis were determined. Animals treated with rFVIIai showed significantly less intimal thickening compared with controls: median 0.147 vs. 0.256 mm2, respectively (p = 0.008). A positive correlation between intimal hyperplasia and medial necrosis (r(s) = 0.79, p = 0.01), as well as adventitial inflammation (r(s) = 0.83, p = 0.009), was found. TF mRNA was not detected in the neointima at 8 weeks, as determined by in situ hybridization. We conclude that active site inhibited FVIIa (rFVIIai) given prior to and directly after implantation of aortic transplants significantly reduces intimal hyperplasia caused by nonimmunological factors in this model. PMID- 12095035 TI - Different mechanisms of cardiac allograft rejection in wildtype and CD28 deficient mice. AB - Although CD28 blockade results in long-term cardiac allograft survival in wildtype mice, CD28-deficient mice effectively reject heart allografts. This study compared the mechanisms of allogeneic responses in wildtype and CD28 deficient mice. Adoptive transfer of purified CD28-deficient T cells into transplanted nude mice resulted in graft rejection. However, this model demonstrated that the allogeneic T cell function was severely impaired when compared with wildtype T cells, despite similar survival kinetics. Cardiac allograft rejection depended on both CD4+ and CD8+ T cell subsets in CD28 deficient mice, whereas only CD4+ T cells were necessary in wildtype recipients. These results suggested that CD8+ T cells were more important in CD28-deficient than wildtype mice. In addition to the CD8+ T cell requirement, allograft rejection in CD28-deficient mice was dependent on a sustained presence of CD4+ T cells, whereas it only required the initial presence of CD4+ T cells in wildtype mice. Taken together, these data suggest that CD4+ T cells from CD28-deficient mice have impaired responses to alloantigen in vivo, thus requiring long-lasting cooperation with CD8+ T cell responses to facilitate graft rejection. These results may help to explain the failure to promote graft tolerance in some preclinical and clinical settings. PMID- 12095036 TI - From first principles--tubulitis in protocol biopsies and learning from history. PMID- 12095037 TI - An analysis of early renal transplant protocol biopsies--the high incidence of subclinical tubulitis. AB - To investigate the possibility that we have been underestimating the true incidence of acute rejection, we began to perform protocol biopsies after kidney transplantation. This analysis looks at the one-week biopsies. Between March 1 and October 1, 1999, 100 adult patients undergoing cadaveric kidney or kidney/pancreas transplantation, or living donor kidney transplantation, underwent 277 biopsies. We focused on the subset of biopsies in patients without delayed graft function (DGF) and with stable or improving renal function, who underwent a biopsy 8.2+/-2.6 d (range 3-18 d) after transplantation (n = 28). Six (21%) patients with no DGF and with stable or improving renal function had borderline histopathology, and 7 (25%) had acute tubulitis on the one-week biopsy. Of the 277 kidney biopsies, there was one (0.4%) serious hemorrhagic complication, in a patient receiving low molecular weight heparin; she ultimately recovered and has normal renal function. Her biopsy showed Banff 1B tubulitis. In patients with stable or improving renal allograft function early after transplantation, subclinical tubulitis may be present in a substantial number of patients. This suggests that the true incidence of rejection may be higher than is clinically appreciated. PMID- 12095038 TI - A study of the pharmacokinetic profile of low-dose hepatitis B immune globulin in long-term liver transplant recipients for chronic hepatitis B infection. AB - Post-transplant protocols for hepatitis B (HBV) prophylaxis using high-dose intravenous hepatitis B immune globulin (10,000 IU) with or without lamivudine are commonly reported. Our centre has previously reported a low-dose intramuscular (i.m.) protocol and lamivudine with excellent results. There have been, however, no pharmacokinetic studies of i.m. hepatitis B immune globulin (HBIG) in this setting. The objective of this study was to determine the pharmacokinetic profile of i.m. HBIG in long-term post-liver-transplant recipients to determine a rational dosing protocol. Six stable liver transplant recipients receiving monthly i.m. HBIG injections for greater than one year were enrolled in this study. All patients had no detectable HBV DNA levels. HBIG titers (anti-HBs) were measured predose, then three times weekly for four weeks and then twice weekly until the serum HBIG titers were 100 IU/L or less. The pharmacokinetic parameters were calculated using noncompartment methods. The mean time to maximum concentration was 10.5 d (range 4-20 d) and the mean half-life was 20 d (range 13.5-23.5 d). Based on these pharmacokinetic parameters in stable long-term post-transplant patients, a rational dosing protocol was developed that allows for more appropriate utility of HBIG and improved patient convenience. PMID- 12095039 TI - Rejection profile of recent pediatric renal transplant recipients compared with historical controls: a report of the North American Pediatric Renal Transplant Cooperative Study (NAPRTCS). AB - Historically, higher acute rejection rates, earlier first rejection, and an inability to reverse the rejection characterize pediatric renal transplantation. In recent years, short-term (1-year) graft survival of pediatric renal transplants has steadily improved. To test the hypothesis that these improvements were mediated by changes in acute rejection, we considered the rejection profile of patients who received a renal allograft between 1987 and 1989 (cohort A) and compared it with recipients transplanted between 1997 and 1999 (Cohort B). Cohort A comprised 1469 transplants and cohort B comprised 1189 transplants. Restricting the data to the first year of follow-up, rejection ratios were 1.6 and 0.7, respectively (p < 0.001). Sixty per cent of the later cohort (B) were rejection free at 1 year, compared with 29% for the earlier cohort (A) (p < 0.001). Controlling for donor source, the rejection reversal rate for the later cohort was significantly better than that of the early cohort (p < 0.001). Cumulative distribution of times to first rejection was significantly better for cohort B (p < 0.001). One-year graft survival for cohort B at 94% was significantly better than 80% for cohort A (p < 0.001). We conclude that the improved short-term graft survival is mediated by improvements in the rejection profile in more recently transplanted patients and that this may translate into a better half-life for pediatric renal transplant recipients who received an allograft in the years 1997 99. PMID- 12095040 TI - In search of the elusive Holy Grail: the mechanisms and prospects for achieving clinical transplantation tolerance. PMID- 12095041 TI - A revised consideration on the use of very aged donors for liver transplantation. AB - The upper age limit for organ donation for liver transplantation has increased over the past few years. A retrospective case control study was carried out to evaluate the outcome of 36 liver transplants (group A) performed with grafts procured from donors over 70 years old in the period 1996 to April 2000, matched with 36 transplants (group B) chronologically performed thereafter with organs procured from donors below the age of 40 yr. The groups were comparable as regards main clinical characteristics. Mean follow-up was 14.5 months. Clinical and laboratory parameters of the donors, cold ischemia period, intraoperative blood transfusions, 30-d mortality, incidence of primary graft nonfunction, acute rejection episodes, arterial complications and long-term survival of recipients were considered. The main postoperative biochemical parameters were also collected and compared. A liver biopsy was obtained in 20/36 old donors, revealing less than 25% of steatosis in all but one, which showed steatosis involving 70% of the hepatocytes. There were two postoperative deaths (5.6%) in group A and one (2.8%) in group B (p = NS). Seven postoperative arterial complications (19.4%) occurred in group A, leading to the patient's death because of rupture of the hepatic artery in one case, to successful surgical revascularization in three cases and to retransplantation in three cases. Only one patient in group B (2.8%) experienced hepatic artery thrombosis (p = 0.055). One-year patient survival rates were 77.4% for group A and 88.8% for group B (p = NS); 1-yr graft survival rates were 73.3% for group A and 85.7% for group B (p = NS). In conclusion, donors over 70 should not be excluded a priori for liver transplantation in elective settings. Great attention should be paid to the pathological conditions of arterial vessels caused by atherosclerosis, i.e. the presence of calcified plaques on the hepatic artery, which might represent the source of severe complications. PMID- 12095042 TI - Effect of extended coverage of immunosuppressive medications by medicare on the survival of cadaveric renal transplants. AB - Between 1993 and 1995, Medicare extended its coverage of maintenance immunosuppression medications following renal transplantation from 1 to 3 years. We hypothesized that Medicare's extension of immunosuppressive coverage would improve graft survival among low-income transplant recipients. We merged patient level clinical data from the USRDS-distributed UNOS registry of kidney transplants throughout the USA with median family income for each patient's ZIP code from the 1990 Census. We were able to merge median incomes to 10,837 first cadaveric renal transplants performed in 1992-93 and 16,732 performed in 1995-97. Each of these chronological cohorts was divided into two groups, those with family incomes above (high-income group) and those below (low-income group) $36,033. There were no differences in graft survival at 1 year based on income in either chronological era. However, when Medicare covered immunosuppression medications for just 1 year, the low-income group of 1-year graft survivors had a 4.5% lower graft survival at the end of 3 years post-transplant (p < 0.001). During the 1995-97 period, during which Medicare provided 3 years' immunosuppression coverage, the low-income and high-income groups had equivalent graft survival at 3 years post-transplant. PMID- 12095043 TI - Attitudes and behavior of young European adults towards the donation of organs--a call for better information. AB - Public perception of organ donation critically affects the availability of organ transplantation in the Western world. To assess the attitude of young adults towards the donation of organs and to investigate potential factors influencing their knowledge and actual behavior regarding organ transplantation, we evaluated a handout questionnaire survey of all Swiss-Italian recruits during six of the years 1989-98 (n = 7272). The attitude of recruits towards organ donation did not change significantly within the 10-year survey period: 61% of young men would personally donate their organs in the case of brain death, 13% would refuse, and 26% had not made up their mind. If they had to decide for close relatives, 50% would consent; 60% of recruits neither knew their next of kin's attitude nor had informed them about their own opinion; 80% felt they were insufficiently informed about organ transplantation. A significantly more positive attitude towards organ donation was found among men who felt they were sufficiently informed, who had close next of kin who were aware of their personal attitude (p < 0.0001), who had contacts with transplanted persons (p < 0.015), or who believed in an existence after death (p < 0.001; chi2-test). Our results suggest that there is potentially large support towards organ donation in this population. To minimize the high rate of indecisiveness, young adults need more appropriate information on the subject and they ask for it. PMID- 12095044 TI - Serial protocol biopsies to quantify the progression of chronic transplant nephropathy in stable renal allografts. AB - AIM: To evaluate the utility of intimal thickness and interstitial width as a primary efficacy variable in the design of clinical trials aimed to modify the natural history of chronic allograft nephropathy. METHODS: A donor and a 4-month protocol biopsy were evaluated in 40 stable grafts according to the Banff schema. In 27 patients, a second protocol biopsy was done at 1 yr. Arterial intimal volume fraction (Vvintima/artery) and cortical interstitial volume fraction (Vvinterstitium/cortex) were estimated with a point counting technique. RESULTS: Chronic Banff scores increased during follow-up, while acute scores reached its peak at 4 months. Vvintima/artery and Vvinterstitium/cortex significantly increased at 4 months, but not at 1 yr. Vvintima/artery at 4 months correlated with donor Vvintima/artery (r = 0.57, p < 0.001), histocompatibility (r = 0.38, p = 0.01) and serum cholesterol (r = 0.31, p = 0.047). Vvinterstitium/cortex at 4 months correlated with recipient body surface area (r = 0.44, p = 0.004) and delayed graft function (p = 0.016). Power calculations showed that Vvintima/artery and Vvinterstitium/cortex allow an important reduction in minimum sample size of a hypothetical trial aimed to prevent chronic allograft nephropathy. CONCLUSIONS: Intimal thickening and interstitial widening progresses rapidly during the first 4 months after transplantation and slowly thereafter. These parameters can be considered as a primary efficacy variable in trials aimed to prevent chronic allograft nephropathy. PMID- 12095045 TI - Simultaneous heart and kidney transplantation in patients with end-stage heart and renal failure. AB - Combined simultaneous organ transplantation has become more common as selection criteria for transplantation have broadened. Broadening selection criteria is secondary to improved immunosuppression and surgical techniques. The kidney is the most common extrathoracic organ to be simultaneously transplanted with the heart. A series of 13 patients suffering from both end-stage heart and renal failure underwent 14 simultaneous heart and kidney transplantations at Temple University Hospital between 1990 and 1999. This is the largest series reported from a single center. Three patients died during the initial hospitalization for an in-hospital mortality of 21%. Of 10 patients who left the hospital, 1-year survival was 100% and 2-year survival 75%. One patient required retransplant for rejection within the first year. Overall mortality at 1 and 2 years was 25 and 41%, respectively. Four out of nine (44%) patients greater than 5 years post transplant were alive. Of the 10 patients who left the hospital, 66% were alive at 5 years. One patient succumbed to primary nonfunction of the cardiac allograft, while the four other deaths were secondary to bacterial or fungal sepsis. The patient's racial backgrounds were equally divided between African American and white. These results are similar to those reported in a United Network of Organ Sharing Database (UNOS) registry analysis of 84 simultaneous heart and kidney transplants that found 1- and 2-year survival to be 76 and 67%, respectively. Simultaneous heart and kidney transplantation continues to be a viable option for patients suffering from failure of these two organ systems, although the results do not match those of heart transplant alone. PMID- 12095047 TI - Addressing minority issues in renal transplantation: is more equitable access an achievable goal? PMID- 12095046 TI - Fatal disseminated aspergillosis following sequential heart and stem cell transplantation for systemic amyloidosis. AB - Infectious complications are a major cause of morbidity and mortality in transplant recipients. We describe a case of fatal disseminated aspergillosis immediately following autologous peripheral stem cell reconstitution in a patient who had undergone orthotopic heart transplantation for systemic amyloidosis. The case described suggests that the infectious risks in patients undergoing these sequential procedures may be distinct from those occurring in patients undergoing either procedure independently. Potential prophylactic and therapeutic interventions are discussed. Since this experimental and evolving approach for the management of systemic amyloidosis is potentially applicable to a limited number of patients, multicenter collaboration may be needed to further define the infectious risks in this unique subset of transplant recipients. PMID- 12095048 TI - Cyclosporine microemulsion and tacrolimus are associated with decreased chronic allograft failure and improved long-term graft survival as compared with sandimmune. AB - Tacrolimus and cyclosporine in the microemulsion formulation Neoral have demonstrated improvements in acute rejection rates after renal transplantation compared with conventional cyclosporine formulation, Sandimmune. To evaluate whether these drugs are also associated with improvements in chronic allograft failure (CAF) rates, we retrospectively analyzed 32,040 primary renal allograft recipients reported to the United States Renal Data System (USRDS) between 1994 and 1997. Graft loss secondary to CAF was defined as graft loss beyond 6 months post-transplant, censored for death, acute rejection, thrombosis, infections and noncompliance. A Cox proportional hazard model was used to investigate the relationship between graft loss secondary to CAF and the use of conventional cyclosporine formulation, as opposed to cyclosporine microemulsion and tacrolimus (Prograf). The analysis was corrected for confounding variables, such as acute rejection, sex, race, human leukocyte antigen (HLA) mismatch, % panel reactive antibodies (PRA), delayed graft function (DGF), cold ischemia time, induction therapy, dialysis time, etiology of end-stage renal disease, cytomegalovirus (CMV) risk group, donor source, era effect, and mycophenolate mofetil (MMF) use. Cyclosporine microemulsion use was associated with a significantly lower relative risk (RR = 0.6, Cl = 0.5-0.7) for CAF as opposed to conventional cyclosporine formulation. Likewise tacrolimus as compared with conventional cyclosporine formulation was associated with a significantly lower relative risk (RR = 0.7, CI = 0.6-0.8) for CAF. Conventional cyclosporine formulation treatment was associated with a 87.6% adjusted CAF-free survival rate at 4 years. Both tacrolimus and cyclosporine microemulsion were associated with a significantly better adjusted CAF-free survival at 4years (91.4 and 92.4%, respectively). Both cyclosporine microemulsion and tacrolimus are associated with improved graft survival and a decreased relative risk for CAF when compared with the older conventional cyclosporine formulation. This association is independent of the use of MMF or changes in era. PMID- 12095049 TI - Positron-emission tomography used to diagnose tuberculosis in a renal transplant patient. AB - We describe the case of a 22-year-old Portuguese renal transplant patient whose post-transplant course was complicated by prolonged delayed graft function and pyrexia of unknown origin. Conventional imaging techniques were not definitive, but positron-emission tomography (PET) scanning identified abnormalities in the lung and mediastinum that led to a diagnostic biopsy demonstrating mycobacterial infection. PMID- 12095050 TI - The role of TNF receptor and TNF superfamily molecules in organ transplantation. AB - The rapid increase in the number of molecules demonstrated to regulate immune responses has provided new opportunities for manipulation of the recipient immune response to transplanted organs. Molecules belonging to the TNF receptors and TNF superfamily are increasingly recognized as playing a major role in the regulation of immune responses to tumor, viral, and autoantigens. The mechanisms by which these molecules regulate immune responses are diverse. TNF receptor-related molecules have been shown to control the development of secondary lymphoid organs, affect the activation and survival of T cells and antigen presenting cells, and alter cytokine and chemokine production. An increasing amount of data suggest that some TNFR superfamily members are particularly important for the function of CD8+ T cells. Based on our current understanding of these molecules it seems highly likely that strategies that target selected TNFR/TNF superfamily molecules will be useful for controlling or preventing the rejection of transplanted organs and tissues. PMID- 12095051 TI - Steroid-free immunosuppression in kidney transplantation: an editorial review. PMID- 12095052 TI - Polyomavirus BK nephropathy: a (re-)emerging complication in renal transplantation. AB - Persisting polyomavirus replication is now widely recognized as a (re-)emerging cause of renal allograft dysfunction. Up to 5% of renal allograft recipients can be affected about 40weeks (range 6-150) post-transplantation. Progression to irreversible failure of the allograft has been observed in up to 45% of all cases. The BK virus strain is involved in the majority of the cases. Risk factors may include treatment of rejection episodes and increasing viral replication under potent immunosuppressive drugs such as tacrolimus, sirolimus or mycophenolate. The diagnosis requires the histological demonstration of nuclear polyomavirus inclusions in affected tubular epithelial cells. Interstitial inflammatory infiltrates and fibrosis become more prominent in the persisting disease and may be difficult to distinguish from (coexisting) rejection. Detection of polyomavirus-inclusion bearing cells ('decoy cells') in the urine and quantification of BK virus DNA in the plasma have been proposed as surrogate markers for polyomavirus replication and allograft disease, respectively. Antiviral treatment is not yet established; however, reports of treatment with cidofovir are encouraging. Current management aims at the judicious modification and/or reduction of immunosuppression which, in view of preceding or concurrent rejection, is not without risk. PMID- 12095054 TI - A tribute to Thomas E. Starzl, MD, PHD. PMID- 12095053 TI - Effect of anti-IL-2Ralpha antibody on IL-2-induced Jak/STAT signaling. AB - Acute allograft rejection is driven by production of cytokines such as interleukin-2 (IL-2) that activate and expand alloreactive T cells by ligating high-affinity IL-2 receptors composed of three subunit chains: alpha, beta, gamma The alpha chain, expressed only on activated T cells, has become an important therapeutic target. Monoclonal antibodies (mAbs) that bind IL-2Ralpha chains significantly decrease transplant rejection. We examined the ability of the humanized anti-IL-2Ralpha antibody daclizumab to block high-affinity IL-2Rs and interrupt T-lymphocyte signaling. Our evaluation focused on a pathway critical for T-cell proliferation, the Jak/STAT pathway. Daclizumab markedly inhibited phosphorylation of the Jak1, Jak3 and STAT5a/b components of the IL-2R-dependent pathway. Suppression by daclizumab was associated with internalization of IL 2Ralpha but not IL-2Rbetagamma chains. High IL-2 doses overcame daclizumab induced blockade of Jak/STAT phosphorylation despite absent cell surface highaffinity IL-2Rs. Under these circumstances, IL-2-mediated Jak/STAT pathway activation might be generated through residual intermediate affinity IL 2Rbetagamma receptors, and this was demonstrated by complete blockade of signaling when anti-IL-2Rbeta monoclonal antibody was added. Humanized antibodies are an important part of strategies to induce alloantigen tolerance. Understanding the molecular events associated with their beneficial clinical effect is critical to design of future immunosuppressive strategies. PMID- 12095055 TI - CTLA-41g in combination with anti-CD40L prolongs xenograft survival and inhibits anti-gal ab production in GT-Ko mice. AB - The generation of GT-Ko mice has provided unique opportunities to study allograft and xenograft rejection in the context of anti-alpha1,3-Gal antibody (anti-Gal Ab) responses. In this study we used the allotransplantation model of C3H hearts into galactosyltransferase-deficient (GT-Ko) mice and the xenotransplantation model of baby Lewis rat hearts into GT-Ko mice to investigate the ability of CTLA 41g in combination with anti-CD40L mAb to control graft rejection and anti-Gal Ab production. Murine CTLA-41g or anti-CD40L monotherapy prolonged allograft survival, and the combination of these reagents was most immunosuppressive. However short-term treatment with murine cytotoxic T lymphocyte associated antigen-4 (muCTLA-41g) and/or CD40 ligand (CD154) monoclonal antibodies (anti CD40L mAbs) was unable to induce indefinite allograft survival. CTLA-4 immunoglobulin fusion protein (CTLA-41g) or anti-CD40L monotherapy only marginally prolonged xenograft survival; the combination of human CTLA-41g and anti-CD40L significantly prolonged xenograft survival (74days), while the combination of murine CTLA-41g and anti-CD40L resulted in graft survival of >120days. CTLA-41g or anti-CD40L monotherapy or the combination of these agents inhibited the production of alloAbs, including anti-Gal Abs. CTLA-41g or anti CD40L monotherapy partially controlled xenoAb and anti-Gal Ab production, while the combination was more effective. These observations corroborate our previous observations that humoral, including anti-Gal Ab, responses and rejection following allograft or concordant xenograft transplantation in GT-Ko mice are T cell dependent and can be controlled by costimulation blockade. PMID- 12095056 TI - Immunoprophylaxis with basiliximab compared with antithymocyte globulin in renal transplant patients receiving MMF-containing triple therapy. AB - Acute graft rejection remains a major problem in renal transplant recipients, and there is no consensus on the optimal immunosuppressive strategy. Immunoprophylaxis with Thymoglobulin or basiliximab has significantly reduced the incidence of acute rejection episodes and graft loss following kidney transplantation. This open, randomized, multicenter study investigated the efficacy and tolerability of basiliximab (20mg day 0-day 4) plus early cyclosporine from day 0 (n = 50) compared with Thymoglobulin plus delayed cyclosporine (n = 50) in adult kidney transplant patients. In addition, all patients received steroids and mycophenolate mofetil (MMF) at standard doses from day 0. Patient and graft survival rates at 12 months were 98 and 94% in the basiliximab group, respectively, compared with 100 and 96% in the Thymoglobulin' group. The incidences of biopsy-confirmed acute rejection (8.0% in each group) and treatment failure (14% in the basiliximab group vs. 8% in the Thymoglobulin group) were comparable in the two groups. There was a non-significant tendency to more dialysis (14 vs. 6%), and fewer cytomegalovirus (CMV) infections (p = 0.005) in the basiliximab group, but the percentage of clinical CMV was not different between the two groups (6 vs. 12%). Both strategies give excellent results, despite the differences in patterns, in nonhyperimmunized patients receiving their first cadaveric renal allograft. PMID- 12095057 TI - Thyroid hormone in the treatment of post-transplant acute tubular necrosis (ATN). AB - Delayed graft function (DGF) in cadaver kidney transplants is a common problem and is often due to acute tubular necrosis (ATN). DGF in transplants may have a deleterious effect on long-term graft survival. Since thyroid hormone has been shown to hasten recovery from ATN in experimental models, we designed a trial to determine if a defined course of triiodothyronine (T3) would improve the short- or long-term outcome of patients with DGF in cadaveric transplants. A prospective, randomized, placebo controlled, double blind trial of T3 was carried out in patients with DGF in cadaveric renal transplants. End-points were percentage requiring dialysis, percentage recovering function, time to recovery and length of hospital stay. Long-term outcomes were percentage grafts functioning at 1 year and mean serum creatinine at 1 year. Forty-four patients were randomized to receive either T3 or placebo. Three patients were dropped from each group when early biopsies disclosed that DGF was due to rejection. The groups were well matched by age, cold ischemia time of the graft, and percentage reactivity to a random panel of antigens. Baseline thyroid function studies, including T3, reverse T3 (rT3), and thyroid stimulating hormone (TSH) levels, were similar between the two groups and typical of 'euthyroid-sick syndrome'. T3 had no effect on percentage requiring dialysis, time to recovery, percentage recovering function, or length of stay. At 1 year follow-up, graft function was similar in both groups and significantly lower than that seen in patients with good initial function. Thyroid hormone, given early in the course of DGF in cadaver kidney recipients, had no effect on the course of DGF. Long-term graft function is impaired in patients who experience post-transplant DGF compared to those who have good initial function. PMID- 12095058 TI - Treatment of osteoporosis and osteopenia in long-term renal transplant patients with alendronate. AB - Bone mineral density (BMD) and biochemical markers of bone-turnover were evaluated in a 2-year study in 58 long-term renal transplant recipients with good renal function. In the first year of study, data were collected and patients with osteoporosis and parameters of high bone turnover were classified as being at high risk for on-going bone loss (Group A; n = 29). Patients with lesser degrees of bone loss or without biochemical parameters of high bone turnover were followed longitudinally (Group B; n = 29). Group A patients were then placed on alendronate 10mg/day and both groups were followed for an additional year. Changes in regional BMD and bone-turnover markers between the first and second year within each group were analyzed using paired tests. BMD in Group A, which had declined at the lumbar spine (- 1.6 +/- 0.5%) and total femur (-1.5 +/- 0.4%) during the first year of the study, increased on alendronate therapy at both the lumbar spine (+3.4 +/- 0.6%, p = 0.001) and total femur (+1.6 +/- 0.6%, p <0.001). These patients also experienced a significant decline in levels of serum alkaline phosphatase, osteocalcin, urinary levels of deoxypyridinoline and pyridinoline. In contrast, neither BMD nor biochemical markers changed significantly over 2 years in Group B. The current results demonstrate that renal transplant patients with osteoporosis and biochemical parameters of high bone turnover are at continued risk for bone loss. Therapy with a bisphosphonate can reverse this bone loss and even increase bone mass in these patients. Whether patients with lesser degrees of bone loss and/or patients without parameters of high bone turnover can also benefit from bisphosphonate therapy deserves further study. PMID- 12095059 TI - Effect of donor factors on early graft survival in adult cadaveric renal transplantation. AB - Previous studies of the effect of donor factors on renal transplant outcomes have not tested the role of recipient body mass index, donor/recipient weight ratios and age matching, and other factors. We analyzed 20,309 adult (age 16 or older) recipients having solitary cadaveric renal transplants from adult donors from 1 July 1994 to 30 June 1998 in an historical cohort study (the 2000 United States Renal Data System) of death censored graft loss by the Cox proportional hazards models, which corrected for characteristics thought to affect outcomes. The only independently significant findings in Cox Regression analysis were a high donor/ recipient age ratio (> or = 1.10, e.g. a 55-year-old donor given to a recipient age 50years or younger, adjusted hazard ratio (AHR) 3.22, 95% confidence interval (CI) 2.36-4.39) and African American donor kidneys (AHR 1.64, 95% CI, 1.24-2.17). African American recipients and older donors were not at independently increased risk of graft failure in this model. Among donor factors, older donor kidneys given to younger recipients and donor African American kidneys were independently associated with graft loss in recipients of cadaver kidneys. The task for the transplant community should be to find the best means for managing all donor organs without discouraging organ donation. PMID- 12095060 TI - Highlights of the Festschrift in honor of Thomas E. Starzl, MD, PHD. PMID- 12095061 TI - Interleukin 1 and chronic rejection: possible genetic links in human heart allografts. AB - Chronic rejection is a leading cause of graft loss in thoracic transplant recipients. Studies on the pathogenesis of chronic rejection have suggested a contributory role for certain cytokines and growth factors. The activity of these mediators is subject to genetic variation if a polymorphism alters expression, or function, of the ligand or its receptor. Here we have asked if certain cytokine and growth factor gene polymorphisms correlate with chronic rejection in recipients of thoracic allografts. In a retrospective analysis of 179 recipients of thoracic organ transplants (128 heart; 36 heart-lung; and 15 lung), polymorphisms in 8 genes that influence the inflammatory process, namely IL1B, IL1R1, IL1RN, IL6, IL10, TNFA, TGFB1 and FCGRIIA, were examined. Genotypic data from recipients who had either died or been re-transplanted as a result of chronic rejection (n = 96) were then compared to those of recipients who had a functioning graft for more than 11 years (n=83). In the heart graft recipients, only those polymorphisms that influenced expression of the IL1 receptor antagonist gene had a significant correlation with graft survival, with homozygosity for the IL1RN*1 allele being associated with rejection. The alternative, less frequent IL1RN alleles emerged as genomic predictors of long term allograft survival. This association was especially strong when IL1 region haplotypes were considered, particularly when analysis was confined to heart transplant recipients who had had multiple acute rejection episodes (OR>20). This case-control study indicates that gene polymorphisms which influence IL1 bioactivity also influence the progression of chronic rejection in heart grafts. PMID- 12095063 TI - Increased access to transplantation for blood group B cadaveric waiting list candidates by using A2 kidneys: time for a new national system? AB - Since blood group B end-stage renal disease (ESRD) patients have less access to donor kidneys and a higher minority composition than any other blood group, the United Network for Organ Sharing (UNOS) approved a voluntary national kidney allocation variance to allow organ procurement organizations (OPOs) to preferentially allocate A2 and A2B kidneys to B candidates. The Midwest Transplant Network OPO has preferentially allocated and transplanted kidneys from blood group A2 and A2B donors to our blood group B waiting list candidates for more than 7 years to increase access to kidneys for the B candidates on our OPO wide waiting list. Between 1994 and 2000, a total of 121 blood group B ESRD patients from our OPO-wide cadaveric kidney waiting list were transplanted. Thirty-four per cent (41/121) of those B candidates received either an A2 or an A2B kidney. One- and 5-year graft survival rates for the group of B recipients of A2 or A2B kidneys were 91 and 85% (died with functioning graft [DWFG] censored), respectively, which were not significantly different from those of 91 and 80% for the 80 B recipients of B or O kidneys (Wilcoxon = 0.48; log-rank = 0.55). These data support the national trial for additional OPOs to voluntarily allocate A2 and A2B kidneys preferentially to B waiting list candidates, thus increasing access of blood group B patients to renal transplantation. PMID- 12095062 TI - Four causes of cadaveric kidney transplant failure: a competing risk analysis. AB - Multiple pathological processes are responsible for renal transplant loss. Development of effective prophylactic regimens requires that risks associated with different outcomes be assessed when several processes act together. A multistate competing risk model that derived multiple endpoint probabilities without averaging impacts of covariates over different outcomes or overestimating fractions of survivors was used to estimate the influence of 16 factors simultaneously on four failure types (acute rejection, chronic rejection, death with a functioning graft and all other causes combined). We analyzed 12,413 non Black and 5,121 Black patients undergoing primary cadaveric kidney transplantation reported to UNOS during 1994-96. Black recipients generally demonstrated higher rates of failure, but the method provided joint probabilities for each failure type and covariate level. To illustrate, a middle-aged Black recipient had probabilities of 6.5%, 5.9%, 2.6% and 6.6% to experience acute rejection failure, chronic rejection failure, death with a functioning graft and other failures, respectively, within 3 years. The corresponding non-Black's chances were 2.9%, 2.5%, 2.7% and 3.6%. The overall 3-year survival equaled 78.4% [100%-(6.5% + 5.9% + 2.6% + 6.6%)] and 88.3% for Blacks and non-Blacks, respectively. The growing complexity of transplantation will benefit from new analytic approaches that accommodate multiple transitions along diverse paths to eventual graft loss or death. PMID- 12095064 TI - Once-daily tazarotene 0.1 % gel versus once-daily tretinoin 0.1 % microsponge gel for the treatment of facial acne vulgaris: a double-blind randomized trial. AB - The efficacy and tolerability of tazarotene 0.1% gel and tretinoin 0.1% microsponge gel were evaluated in a multicenter, double-blind, randomized, parallel-group study in patients with mild-to-moderate inflammatory facial acne vulgaris. A total of 169 patients were randomized to once-daily applications of one of these topical retinoids for 12 weeks. Both agents were associated with significant reductions from baseline in the noninflammatory and inflammatory lesion counts. Tazarotene treatment was associated with a significantly greater incidence of treatment success (defined as > or = 50% global improvement [67% vs 49%; P=.03]) and significantly greater reductions in overall disease severity (36% vs 26%; P=.02) and noninflammatory lesion count (60% vs 38% at week 12; P=.02) than tretinoin microsponge treatment. Both drugs were well tolerated, with mean levels of dryness, burning, pruritus, erythema, and peeling generally being no more than trace throughout the study. There were no clinically significant between-group differences in these measures of tolerability. Two patients in each group (2%) discontinued because of treatment-related adverse events. The mean amount of medication applied by the patients was 0.28 g per application with tazarotene and 0.41 g per application with tretinoin microsponge, resulting in cost-effectiveness ratios of $81.45 per treatment success with tazarotene and $108.24 per treatment success with tretinoin microsponge. Tazarotene was observed to have greater efficacy and comparable tolerability and to be a cost-effective alternative to tretinoin 0.1% microsponge gel. PMID- 12095066 TI - A multicenter, double-blind, randomized comparison study of the efficacy and tolerability of once-daily tazarotene 0.1% gel and adapalene 0.1% gel for the treatment of facial acne vulgaris. AB - The efficacy and tolerability of tazarotene 0.1% gel and adapalene 0.1% gel were compared in a multicenter, double-blind, randomized, parallel-group study in 145 patients with mild-to-moderate facial acne vulgaris. Both treatments were applied once daily in the evenings for up to 12 weeks. Compared with adapalene, treatment with tazarotene was associated with a significantly greater incidence of treatment success (> or = 50% global improvement) (78% vs 52%; P=.002) and significantly greater reductions in overall disease severity (P<.0001), noninflammatory lesion count (P<.0001), and inflammatory lesion count (P=.0002). In the early weeks of treatment, tazarotene was associated with transiently greater levels of burning, pruritus, erythema, and peeling compared with adapalene (P<.01). However, mean levels of these parameters were consistently less than mild in both treatment groups and, at the end of treatment, patients considered both treatments to be comparably well tolerated (the proportion of patients in each group who rated the comfort of their treated skin as comfortable or very comfortable was 76% with tazarotene and 69% with adapalene). Mean usage of study medication was 0.32 g per application of tazarotene and 0.42 g per application of adapalene, which resulted in cost-effectiveness ratios of $79.95 per treatment success for tazarotene and $107.88 per treatment success for adapalene. Sensitivity analyses suggest that these cost-effectiveness results are robust across a range of cost and efficacy assumptions. In conclusion, tazarotene 0.1% gel was more effective than adapalene 0.1% gel and was also a more cost effective treatment option. PMID- 12095067 TI - Aminolevulinic acid hydrochloride with photodynamic therapy: efficacy outcomes and recurrence 4 years after treatment. AB - The safety and efficacy of treating individuals who presented with multiple actinic keratosis (AK) lesions with 5-aminolevulinic acid (ALA) in combination with photodynamic therapy (PDT) were documented in a phase III trial. This report highlights results of this phase III trial and reviews 4 specific cases of sustained AK lesion clearance 4 years after treatment with ALA/PDT Long-term recurrence data were collected from patients who participated in clinical trials of ALA/PDT Long-term evaluation extended to 36 to 48 months (4 years) supports primary efficacy findings of the phase III pivotal trial, with a low incidence of AK recurrence in patients treated with ALA/PDT PMID- 12095065 TI - Optimizing the use of tazarotene for the treatment of facial acne vulgaris through combination therapy. AB - A multicenter, investigator-masked, randomized, parallel-group study was performed in 440 patients with mild-to-moderate facial acne vulgaris to compare the efficacy and tolerability of tazarotene monotherapy with 3 combination regimens--tazarotene plus benzoyl peroxide gel, tazarotene plus erythromycin/benzoyl peroxide gel, and tazarotene plus clindamycin phosphate lotion. An additional treatment group-monotherapy with clindamycin phosphate lotion-also was included as a reference arm. The only combination therapy to achieve a significantly greater global improvement than tazarotene monotherapy was tazarotene plus clindamycin. For reducing noninflammatory lesions specifically, none of the combination regimens offered significant benefit over tazarotene monotherapy (though tazarotene plus clindamycin and tazarotene plus erythromycin/benzoyl peroxide were significantly more efficacious than clindamycin monotherapy). For reducing inflammatory lesions, tazarotene plus erythromycin/benzoyl peroxide was significantly more efficacious than all the other regimens. Although tazarotene plus clindamycin and tazarotene plus benzoyl peroxide reduced the incidence of adverse effects compared with tazarotene monotherapy, the difference did not achieve statistical significance. PMID- 12095068 TI - The evolving role of aminolevulinic acid hydrochloride with photodynamic therapy in photoaging. AB - The safety and efficacy of treating individuals who presented with multiple actinic keratosis (AK) lesions with 5-aminolevulinic acid (ALA) in combination with photodynamic therapy (PDT) were documented in a phase III trial. This report highlights results of this phase III trial and reviews 2 specific case studies treated with ALA/PDT who presented with both multiple nonhyperkeratotic AK lesions and moderate-to-severe photodamage. Treatment consisted of a 2-step process performed by the investigator. In addition to the pretreatment evaluation, each subject was evaluated at 1- and 2-month intervals after each treatment of ALA/PDT. Clinical findings reveal complete healing at the site of ALA/PDT without scarring or changes in pigmentation, as well as significant changes in signs of photodamage, such as improvement in skin elasticity and reduction in skin thickening. PMID- 12095069 TI - Reduction in preference for saccharin by repeated unpredictable stress in mice and its prevention by imipramine. AB - The present study set out to establish the chronic mild stress (CMS) animal model of depression in male CD-1 mice, a commonly used mouse strain. Mice were exposed to a series of mild stressors (e.g. soiled bedding, paired housing, cage tilt, white noise) presented in a continuous unpredictable fashion. Intermittently, CMS was discontinued and the mice were presented with both water and a palatable saccharin solution (0.1% w/v) in a two-bottle choice test overnight (15 h). Repeated exposure of these mice to the stressors led to a reduction in preference for the saccharin solution. This change in preference was attributed to an increase in the consumption of water rather than a decrease in the consumption of saccharin solution. Over time and with extensive testing, CMS no longer affected performance in the two-bottle saccharin preference test. Treatment with the tricyclic antidepressant imipramine (20 mg/kg i.p., once daily) had a varied effect on the CMS-induced change in preference for saccharin, dependent on the timing of initiation of imipramine treatment. In the first instance, following 5 weeks of CMS where a reduction in saccharin preference was established, treatment with imipramine for a further 5 weeks maintained the stress-induced deficit in saccharin preference. However, using a different approach, pre-treatment with imipramine once daily for 2 weeks, prior to onset of CMS, and co-treatment thereafter, attenuated CMS-induced changes in saccharin preference. Finally, when imipramine treatment was scheduled to begin with the CMS procedure, imipramine failed to prevent the CMS-induced reductions in saccharin preference. Changes in behaviour observed after exposure to CMS may be linked to a stress-induced deterioration of the sensitivity of the mice to a rewarding stimulus. Treatment with imipramine can reduce these behavioural changes but is only effective when given repeatedly prior to onset of CMS. PMID- 12095070 TI - Inhibition by venlafaxine of the increase in norepinephrine output in rat prefrontal cortex elicited by acute stress or by the anxiogenic drug FG 7142. AB - Venlafaxine is an antidepressant drug that inhibits the reuptake of serotonin and norepinephrine with different efficacies. The effects of repeated administration of this drug on the increase in the extracellular concentration of norepinephrine in the prefrontal cortex, induced by stress or by the anxiogenic drug FG 7142, were studied in freely moving rats. Exposure to foot-shock stress induced a marked increase (+120%) in the extracellular norepinephrine concentration in the prefrontal cortex of control rats. Long-term administration of venlafaxine (10 mg/kg i.p., once a day for 21 days) reduced the effect of stress on norepinephrine output by 75%. This effect of venlafaxine persisted for at least 5 days after discontinuation of drug treatment. Acute administration of FG 7142 (20 mg/kg i.p.), a benzodiazepine receptor inverse agonist, increased norepinephrine output (+90%) in control rats. Chronic treatment with venlafaxine prevented the effect of FG 7142. In contrast, the acute administration of this antidepressant had no effect on the stress- or FG 7142-induced increase in norepinephrine output. These plastic changes in the sensitivity of norepinephrine neurones to foot-shock stress and to an anxiogenic drug may reveal an important neuronal mechanism for the physiological regulation of emotional state. Furthermore, this mechanism might be relevant to the anxiolytic and antidepressant effects of venlafaxine. PMID- 12095071 TI - Prevention of the stress-induced increase in the concentration of neuroactive steroids in rat brain by long-term administration of mirtazapine but not of fluoxetine. AB - The effects of acute and chronic administration of fluoxetine on the basal and stress-induced increases in cerebrocortical and plasma concentrations of allopregnanolone (3alpha,5alpha-tetrahydroprogesterone; 3alpha,5alpha-TH PROG) and tetrahydrodeoxycorticosterone (3alpha,5alpha-TH DOC) were compared with those of mirtazapine, an antidepressant that (unlike fluoxetine) is not a selective serotonin reuptake inhibitor. A single injection (20 mg/kg i.p.) of fluoxetine or mirtazapine resulted in significant increases in the cerebrocortical and plasma concentrations of 3alpha,5alpha-TH PROG and 3alpha,5alpha-TH DOC. In contrast, long-term administration (10 mg/kg i.p., once daily for 2 weeks) of fluoxetine, but not that of mirtazapine, induced marked decreases in the cortical and plasma concentrations of these neuroactive steroids. Chronic treatment with fluoxetine, however, did not inhibit the increases in the cortical and plasma concentrations of 3alpha,5alpha-TH PROG and 3alpha,5alpha-TH DOC induced by acute foot-shock stress. In contrast, chronic treatment with mirtazapine prevented or significantly reduced the stress-induced increases in neurosteroid concentrations in the cerebral cortex and plasma, respectively. These results show that mirtazapine, similar to fluoxetine, initially increases the cortical concentration of neuroactive steroids; however, chronic administration of this drug modulates the plasma and brain availability of these hormones in a manner distinct from that of fluoxetine. PMID- 12095072 TI - Inhibition of methamphetamine self-administration in rats by cannabinoid receptor antagonist AM 251. AB - Cannabinoids are drugs that are frequently abused not only alone, but also in combination with other drugs. The present study investigated possible functional interactions between the psychostimulant methamphetamine and the cannabinoid receptor agonists anandamide, or R-(+)-methanandamide and cannabinoid antagonist AM 251 in the rat model of i.v. drug self-administration. In rats trained to self administer methamphetamine, the intake was significantly decreased by the cannabinoid antagonist and tended to be dose-dependently increased by pre treatment with cannabinoid receptor agonists. Possible mechanisms for these drug interactions are discussed and the use of the cannabinoid antagonist for the treatment of drug abuse is considered. PMID- 12095073 TI - Effect of chronic fluoxetine and WAY-100635 treatment on serotonergic neurotransmission in the frontal cortex. AB - Clinical augmentation strategies have shown that some improvement in antidepressant efficacy can be achieved by combining the beta adrenergic/serotonin (5-HT)1A/1B receptor antagonist (+/-)pindolol with a selective serotonin reuptake inhibitor (SSRI). This has lead to the hypothesis that a combination of a 5-HT1A receptor antagonist with an SSRI will lead to a faster onset of antidepressant action. Although there is a significant accumulation of acute preclinical data supporting this rationale, until recently, there have been no investigations examining the chronic effects of combining an SSRI with a 5-HT1A receptor antagonist. Here, we determined the chronic effects of fluoxetine (10 mg/kg o.d.), administered in combination with the selective 5 HT1A receptor antagonist WAY-100635 (1 mg/kg b.i.d.), on serotonergic neurotransmission in the frontal cortex using in-vivo microdialysis. Following chronic administration of fluoxetine +/- WAY-100635, functional changes in serotonergic neurotransmission, as well as 5-HT1A autoreceptors, were assessed by administering fluoxetine or (+/-) 8-hydroxy-2-(di-n-propylamino)tetralin [(+/-) 8 OH-DPAT] 24 h after the last chronic dose. Chronic administration of WAY-100635 alone produced no detectable change in the functional status of the 5-HT1A receptor. However, fluoxetine alone produced a time-dependent adaptation in serotonergic transmission such that fluoxetine (acutely administered on day 15) was able to produce a two-fold increase in extracellular 5-HT levels but the decrease in response to 8-OH-DPAT was completely attenuated. These data indicate that the fluoxetine-induced adaptation was mediated by desensitization of the 5 HT1A receptor. WAY-100635 given chronically in combination with fluoxetine blocked the SSRI-induced desensitization of the 5-HT1A receptor. Furthermore, chronic treatment with this combination produced no tolerance in terms of its ability to acutely increase forebrain 5-HT levels. These data suggest that augmentation of an SSRI by combined pharmacotherapy with a 5-HT1A antagonist would be effective upon prolonged exposure. PMID- 12095074 TI - Nimodipine prevents scopolamine-induced impairments in object recognition. AB - The effects of acute administration of the dihydropyridine calcium channel antagonist, nimodipine, were studied on the actions of scopolamine in the object recognition test. Scopolamine at 0.125 mg/kg decreased the difference in the time spent exploring novel and familiar objects when given either 15 min before, or immediately after, exposure to objects. Administration of nimodipine at 10 mg/kg, or 1 mg/kg, at the same time as the scopolamine completely prevented the deleterious effects on memory in this task. This effect was seen when nimodipine and/or scopolamine were given prior to the object exposure and also when the drugs were given after the experience of seeing the objects. Nimodipine had no effects on performance when given in the absence of scopolamine. This lack of change in total time spent exploring the objects indicated that the effects of scopolamine and nimodipine were not due to changes in motor coordination or alertness. The results are discussed in the light of the role of cholinergic transmission in memory and the known actions of dihydropyridines on brain function. PMID- 12095076 TI - The cost-effectiveness of clozapine: a controlled, population-based, mirror-image study. AB - A retrospective cohort study, with a mirror-image design, was used to measure inpatient service utilization in 63 consecutive patients started on clozapine from a geographical catchment area compared to a control group matched for previous inpatient service use. An intent-to-treat analysis, including those patients (n = 28) who discontinued clozapine during the study period, showed a significant reduction in number of admissions and total time spent in hospital in the 2 years following clozapine initiation compared to the previous 2 years and to the follow-up period in the control group. This translated into a reduction of 7,300 pounds in hospitalization costs per patient started on clozapine, over the 2-year period. In those patients who continued clozapine treatment for the whole of the 2-year period, there was a two-thirds reduction in number of admissions and total time spent in hospital compared to no change in the clozapine discontinuers. These findings suggest that clozapine is a clinically and cost effective intervention for severe schizophrenia in routine clinical settings. PMID- 12095075 TI - Cocaine-primed craving and its relationship to depressive symptomatology in individuals with cocaine dependence. AB - Several lines of evidence suggest a link between cocaine-primed craving and depressive symptomatology. The purpose of this study was to directly relate these two clinical phenomena. Thirty-three cocaine-dependent subjects were rated on the Hamilton Rating Scale for Depression (HRSD) at baseline and then administered an i.v. bolus of cocaine (0.2 mg/kg). Multiple regression analysis revealed that only the HRSD score was an independent predictor of cocaine-primed craving (F= 4.09; d.f. = 10,22; r = 0.81, p < 0.003) when baseline spontaneous craving during early withdrawal, age, gender, frequency of use, time since last use, monetary expenditure on cocaine and the Addiction Severity Index Drug Composite Scores were considered. These data support the hypothesis that depressive symptomatology affects cocaine-primed craving and that this relationship is relatively specific to symptoms defined by the HRSD and is not seen with a number of other clinical and demographic variables. PMID- 12095077 TI - Side-effects of risperidone therapy mimicking cerebrospinal fluid shunt malfunction: implications for clinical monitoring and management. AB - Risperidone is an atypical antipsychotic used in the treatment of several psychiatric disorders in both children and adults. We present two patients with hydrocephalus and learning difficulties who were admitted to the neurosurgical unit with a suspected cerebrospinal fluid shunt malfunction and raised intracranial pressure. They had both been commenced on risperidone for the treatment of aggressive outbursts. Twelve days after commencing risperidone, the first patient developed symptoms of headache, nausea, vomiting, drowsiness, lethargy and two episodes of collapse. The second patient presented with similar symptoms 4 days after his risperidone dose was increased. An unnecessary shunt exploration was averted in both cases when it was noted that the side-effect profile of risperidone mimicked exactly those of shunt malfunction. Discontinuation of the drug resulted in complete resolution of all symptoms within 72 h. Many patients with shunted hydrocephalus have associated developmental disorders that may warrant treatment with risperidone. Clinicians should be aware of the potential symptom overlap between shunt malfunction and risperidone side-effects in these patients. PMID- 12095078 TI - A possible association between high normal and high dose olanzapine and prolongation of the PR interval. AB - Two cases of light-headedness or 'fainting' in patients with schizophrenia treated with olanzapine are presented. Electrocardiogram investigation revealed first-degree heart block on high normal and high dose olanzapine, which normalized after dose reduction. PMID- 12095079 TI - Effects of 10 mg and 15 mg oral procyclidine on critical flicker fusion threshold and cardiac functioning in healthy human subjects. AB - The critical flicker fusion threshold (CFFT) is thought to index alertness and cortical arousal. Sedative drugs reduce CFFT while psychostimulants increase it. Procyclidine is an anticholinergic that is used to control the extrapyramidal side-effects of antipsychotics in schizophrenia. This study examined the effects of clinically relevant doses of oral procyclidine administration on CFFT and heart rate in two separate experiments (Experiment 1, drug dose: 10 mg, n = 16; Experiment 2, drug dose: 15 mg, n = 12) involving healthy subjects using a double blind, placebo-controlled, cross-over design. 10 mg procyclidine had no significant effect on CFFT, heart rate or self-ratings of mood, but the 15 mg dose significantly lowered CFFT at 1 h and 2 h after procyclidine administration, increased drowsiness ratings and produced a drop in heart rate. The effects observed in this study may have implications for treatment compliance of schizophrenic patients, choice of antipsychotics, prescribing to patients with heart disease and monitoring of cardiac function under treatment. Further investigations are required to quantify the effects of procyclidine on CFFT and cardiac function in patients with schizophrenia. PMID- 12095080 TI - What is a dose of ecstasy? PMID- 12095082 TI - Equine herpesvirus 1 and 4 infections: an update. AB - Equine herpesvirus 1 (EHV1) and equine herpesvirus 4 (EHV4) are important ubiquitous equine viral pathogens, causing much damage to the horse industry. EHV1 strains are associated with respiratory disease, abortion, and paresis/paralysis, whereas EHV4 strains are predominantly associated with respiratory disease. In the past decades much research effort has been put into improving knowledge about these viruses. In this paper the current state of knowledge of these viruses and the most important aspects of these virus infections, e.g. epidemiology, clinical aspects, pathogenesis and pathology, immunity, diagnosis, preventive management and management in the course of an outbreak and vaccination, is reviewed. Because we performed some research ourselves in the areas of diagnosis, epidemiology and vaccinology these aspects are reviewed in more depth than the other aspects. Still many questions have remained and new questions have risen. Consequently, research priorities should be made in an attempt to answer these questions. Therefore, this review ends with some personal recommendations for important priorities for future research. PMID- 12095081 TI - Probable serotonin syndrome variant in a patient receiving a selective serotonin reuptake inhibitor and a 5-HT3 receptor antagonist. PMID- 12095084 TI - Porcine reproductive and respiratory syndrome (PRRS) with special reference to clinical aspects and diagnosis. A review. AB - After a short introduction on Porcine Reproductive and Respiratory Syndrome (PRRS) regarding the history, the first occurrence in several countries, and the causal virus, designated Lelystad virus, a description is given of the clinical aspects and several diagnostic methods. After some general remarks on the clinical aspects, the epidemic and the endemic phase of the disease are described. Regarding the diagnosis, special attention is paid to the detection of antibodies and of the PRRS Virus (PRRSV). Regarding the detection of antibodies, a description is given of three tests: the immunoperoxidase monolayer assay, the enzyme-linked immunosorbent assay, and the serum neutralization test. Concerning the detection of PRRSV, attention is paid to the isolation of the virus, the demonstration of PRRSV antigens in frozen or fixed tissue using immunohistochemistry or immunofluorescence, the in situ hybridisation technique and the Polymerase Chain Reaction (PCR). PMID- 12095083 TI - Equine influenza virus infections: an update. AB - Equine influenza is one of the most economically important contagious respiratory diseases of horses. In this paper the current state of knowledge of equine influenza virus and the most important aspects of these virus infections, e.g. epidemiology, clinical aspects, pathogenesis and pathology, immunity, diagnosis, treatment, management and vaccination, are reviewed with an emphasis on epidemiology, diagnosis and vaccinology. Many questions have remained and with the advent of improved technology new questions have arisen. Consequently, research priorities should be set in an attempt to answer them. Therefore, this review ends with some personal recommendations for important priorities for future research. PMID- 12095085 TI - Facilitating children's understanding of misinterpretation: explanatory efforts and improvements in perspective taking. AB - The authors investigated children's understanding of how mistaken beliefs can arise through misinterpretation of ambiguous information. Children (N = 91), aged 4 to 5 years, were given pre- and posttests on their ability to infer a puppet's interpretation of a restricted-view drawing after the puppet had been led to an erroneous expectation about the drawing's identity. Before the posttest, the children received either self-explanation training or other-explanation training in which they explained the source of their own or a puppet's misinterpretations of drawings; a control group received no training. The children who received training improved from pre- to posttest, and those who had practiced explaining misinterpretations by referring to previously viewed pictures or to features of a target picture showed the greatest improvement. These results indicate that learning to explain misinterpretations can help children recognize situations in which misinterpretations are likely to occur. PMID- 12095086 TI - Thinking styles and academic achievement among Filipino students. AB - The authors' objective in this study was to determine whether the precepts of R. J. Sternberg's (1988, 1997) theory of mental self-government apply to a non Western culture. They administered R. J. Sternberg and R. K. Wagner's (1992) Thinking Styles Inventory, which is based on the theory of mental self government, to 429 Filipino university students. The results of item analysis, scale intercorrelations, and factor analysis were consistent with the general provisions of the theory. Correlational analysis between thinking styles and grade point average showed that thinking styles are related to acade micachievement. The results are explained with respect to the concepts and practices of Philippine culture and schools and discussed in relation to the developmental assumptions of the theory of mental self-government. PMID- 12095087 TI - Chinese and Dutch parents' perceptions of their children's personality. AB - The authors coded Chinese (n = 401) and Dutch (n = 324) parents' free descriptions of their 3- to 14-year-old children's personalities using a 14 category coding system partially based on the Big Five. Of the Chinese and Dutch personality descriptors, 86% and 77%, respectively, could be classified in the first 5 main categories resembling the five-factor model of adult personality. No significant differences were found for gender, socio-economic status, or city in these categories. Chinese parents of school age children generated many more descriptors, mostly critical, in the domain of conscientiousness. The findings reflect Chinese high achievement orientation and show that the classification system, which presently serves as a basis for developing indigenous questionnaires for personality assessment of children in China and some Western countries, is sensitive to cultural differences. PMID- 12095088 TI - Thinking styles and cognitive development. AB - Using R. J. Sternberg's (1988, 1997) theory of thinking styles and W. G. Perry's (1970) theory of cognitive development, the author investigated the nature of thinking styles as they relate to cognitive development. Eighty-two Hong Kong university students (44 male, 38 female) responded to the Thinking Styles Inventory (R. J. Sternberg & R. K. Wagner, 1992) and the Zhang Cognitive Development Inventory (L. F. Zhang, 1997). Statistical analyses provided varying degrees of support for the prediction of an overlap between the thinking style and cognitive development constructs. In general, students who reasoned at a higher cognitive developmental level tended to use a wider range of thinking styles than students who reasoned at a lower cognitive developmental level. Implications of results are discussed in relation to education and research. PMID- 12095089 TI - Gender influences on preschool children's social problem-solving strategies. AB - The authors investigated gender influences on the nature and competency of preschool children's social problem-solving strategies. Preschool-age children (N = 179; 91 boys, 88 girls) responded to hypothetical social situations designed to assess their social problem-solving skills in the areas of provocation, peer group entry, and sharing or taking turns. Results indicated that, overall, girls' responses were more competent (i.e., reflective of successful functioning with peers) than those of boys, and girls' strategies were less likely to involve retaliation or verbal or physical aggression. The competency of the children's responses also varied with the gender of the target child. Findings are discussed in terms of the influence of gender-related social experiences on the types of strategies and behaviors that may be viewed as competent for boys and girls of preschool age. PMID- 12095090 TI - Bartlett revisited: reconfiguration of long-term memory in young and older adults. AB - This study was performed after the tradition of F. C. Bartlett (1932), who demonstrated that memory reconfigures over time. The authors investigated the memory of young and older adults to examine the degree to which the aging process influences reconfigurative tendencies. From an initial sample of 53 participants, 20 young and 19 older adults completed 6 tests of recall for Bartlett's original text materials over an 84-day period. Consistent with the broad conclusions of Bartlett's study, reconfiguration was observed: Both young and older adults introduced errors into memory. Older adult recall was lower overall than that of young adults, and recall performance diminished over time. However, there was no difference between the performances of young and older adults with respect to incorrectly recalled intrusive elements. PMID- 12095091 TI - Firstborn preference and attitudes toward using sex selection technology. AB - The authors investigated sex preferences for firstborn children and (a) pro- technology attitudes and willingness to use sex selection technology (SST) and (b) social conformity. College students (N = 469; 239 women, 230 men) answered demographic questions and questions on technology usage and personal preferences and completed a measure of conformity. Findings revealed a significant overall preference for firstborn males, a specific preference of potential SST users for firstborn sons, and a greater willingness of African American students, compared with White students, to use SST. No correlation was found between conformity and potential SST use. Logistic regression results suggested that protechnology attitudes may be predictive of firstborn preferences. Implications of the findings are discussed in relation to gender equality and population sex ratios. PMID- 12095092 TI - When astronomy, biology, and culture converge: children's conceptions about birthdays. AB - The authors investigated the development of children's understanding of birthdays using structured interviews of 102 Israeli children aged 4 to 9 years. To fully comprehend the concept of birthday, children must grasp the relationship between the social occasion (the birthday party), irreversible biological growth, and the cyclical nature of the calendar. The authors' findings affirmed that a child's early conception is wholly social and self-contained (birthday parties confer a new age) and that young children believe that age can be affected by multiplying or skipping birthdays. The mature conception is socially based, but it is integrated with additional conceptual subsystems: the irreversible and independent unfolding of biological growth and the cyclical aspect of time. This enables the child to go beyond a magical approach to birthday rituals. PMID- 12095093 TI - The big picture. PMID- 12095094 TI - Induction of proliferative vitreoretinopathy by a unique line of human retinal pigment epithelial cells. AB - BACKGROUND: The most widely used models of proliferative vitreoretinopathy (PVR) rely on injection of cells into the vitreous of animals. Using retinal pigment epithelial (RPE) cells from human PVR membranes may produce a more accurate model of human PVR. We performed a study to determine whether human RPE cells derived from a single epiretinal membrane (ERM) are capable of inducing the same disease in the rabbit eye, and whether the induced ERMs had cellular components similar to those of human PVR membranes. METHODS: Cells were harvested from a human ERM obtained at surgery for PVR. RPE cells were cultured from the membrane and injected into the right eye of 24 New Zealand albino rabbits. The left eyes served as controls. The eyes were examined by indirect ophthalmoscopy over 4 weeks. The enucleated eyes were then examined by means of microscopy and histochemical analysis. RESULTS: By day 7, PVR had developed in all but 1 of the 24 experimental eyes, with 8 progressing to localized tractional retinal detachment. By day 21, localized tractional retinal detachment had developed in 17 eyes; 1 eye progressed to extensive tractional retinal detachment by day 28. Immunostaining showed that mostly RPE cells, but also myofibroblasts, glial cells and collagen, were present in the newly formed rabbit PVR membranes. INTERPRETATION: Human RPE cells cultured from a PVR membrane appear to be capable of inducing PVR in rabbits. The resultant ERMs are similar to those formed in human PVR and consist mainly of RPE cells. PMID- 12095095 TI - Membrane conductance of Muller glial cells in proliferative diabetic retinopathy. AB - BACKGROUND: It is not known whether the membrane features of human Muller cells are altered in proliferative diabetic retinopathy (PDR). We performed a study to investigate the expression of several distinct forms of membrane conductance in Muller glial cells from a patient with PDR compared to cells from healthy donors (control cells). METHODS: Muller cells were isolated 2 hours after vitreoretinal surgery in the case of the patient and within 24 hours in the case of the autopsy eyes. Whole-cell voltage-clamp recordings were made. The results for the two groups were compared with the Mann-Whitney U test. RESULTS: As assayed by the whole-cell membrane capacitance, the cells from the patient with PDR showed hypertrophy in comparison to the control cells (mean 85.1 pF [standard deviation (SD) 19.7 pF] vs. 54.3 pF [SD 13.8 pF]). The cells from the patient displayed strong downregulation of inwardly rectifying potassium ion (Kir) currents (mean 0.41 [SD 0.24] pA/pF, compared to 3.43 [SD 1.86] pA/pF for the control cells). The Kir current downregulation was accompanied by a less negative membrane potential (-57.3 mV [SD 16.9 mV], compared with -82.3 mV [SD 5.3 mV] for the control cells). Both the number and the amplitude of voltage-gated sodium ion currents were enhanced in cells from the patient. When P2X7 receptors were activated by 2'-/3'-O-(4-benzoylbenzoyl)-adenosine triphosphate, cells in both groups displayed opening of a cation conductance and, simultaneously, an increase in currents through calcium ion-activated potassium ion channels. INTERPRETATION: Changes in Muller cell membrane conductance in PDR are similar to those described in proliferative vitreoretinopathy. The down-regulation of active Kir channels and the membrane depolarization likely disturb voltage-dependent Muller cell functions, such as regulation of local ion concentrations and uptake of neurotransmitters. The enhanced entry of calcium ions from the extracellular space and the subsequent stimulation of calcium-activated potassium channels support Muller cell proliferation in PDR. PMID- 12095096 TI - Treatment of acute bacterial conjunctivitis: 1% fusidic acid viscous drops vs. 0.3% tobramycin drops. AB - BACKGROUND: A frequent cause of conjunctivitis is an acute bacterial infection, presenting with mucopurulent discharge and conjunctival hyperemia. The authors compared the clinical and microbiologic efficacy, safety and acceptability of 1% fusidic acid viscous drops (Fucithalmic) with 0.3% tobramycin ophthalmic solution (Tobrex) in the treatment of suspected bacterial conjunctivitis. METHODS: Patients were recruited at 20 sites in Ontario, Saskatchewan and Alberta from October 1995 to December 1998. Patients who presented to their primary care physician with suspected bacterial conjunctivitis, as identified by conjunctival hyperemia and purulent or mucopurulent discharge, were eligible for the study. Patients were randomly assigned to receive 7 days of treatment with either 1% fusidic acid (one drop applied twice daily) or 0.3% tobramycin (one to two drops applied four to six times daily). The investigators were blinded as to treatment status. Bacteriologic samples were taken from the inferior conjunctival cul-de sac on day 0 and at the end of treatment. Signs and symptoms of conjunctivitis were assessed at baseline and after 3 and 7 days of treatment. The acceptability of treatment was assessed by having the patient or the parent or guardian complete a questionnaire on degree of compliance and ease of use after 3 and 7 days of treatment. RESULTS: Conjunctival swabs were obtained from 484 patients (410 over 9 years of age and 74 aged 2 to 9 years) to determine baseline bacteriology. Of the 484, 319 (65.9%) (63% of the older patients and 80% of those aged 2 to 9 years) had positive results of culture for bacteria. Ninety-four patients (19%) (63 [15%] of the older patients and 31 [42%] of those aged 2 to 9 years) had per-protocol pathogens as defined by quantitative bacteriology criteria. There was a direct correlation between the presence of mucopurulent discharge and the presence of per-protocol pathogens. There were no significant differences in clinical or bacteriologic efficacy between the treatment groups. Treatment compliance was similar between the treatment groups for the older patients; however, for those aged 2 to 9 years, compliance was significantly better in the fusidic acid group than in the tobramycin group (85% vs. 47%) (p < 0.001). Significantly more patients in the fusidic acid group than in the tobramycin group rated treatment as convenient or very convenient, particularly among younger patients (97% vs. 54%) (p < 0.001). INTERPRETATION: The clinical and bacteriologic efficacy of fusidic acid viscous drops combined with the convenience of a twice-daily dosage regimen establishes this antibiotic as first line treatment for suspected acute bacterial conjunctivitis and a favourable alternative to other broad-spectrum antibiotics. PMID- 12095097 TI - Tarsal strip combined with modified Quickert-Rathbun sutures for involutional entropion. AB - BACKGROUND: Involutional entropion is a common problem in the elderly population. The author describes a method of repair of involutional entropion that is a combination and modification of two existing surgical techniques. METHODS: Review of the cases of 20 consecutive patients (22 lids) who underwent surgery for involutional entropion. All surgical procedures were performed by the author. Repair consisted of a tarsal strip procedure combined with a modification of the Quickert-Rathbun suture technique. All patients had at least 1 year of follow-up. RESULTS: Surgery was successful in 21 (95%) of the 22 eyelids. One patient was noted to have recurrent entropion at 21 months. The average length of follow-up was 33.3 months (range 12 to 79 months). INTERPRETATION: The surgical technique described is straightforward and reliable for the correction of involutional entropion. PMID- 12095098 TI - Monocanalicular lacerations: to reconstruct or not? PMID- 12095099 TI - Melanocytoma of the choroid: atypical presentation and review of the literature. PMID- 12095100 TI - Eye injuries in Canadian sports and recreation, 1972-2002. PMID- 12095101 TI - The effects of supraoptimal temperatures on population growth and cortical patterning in Tetrahymena pyriformis and Tetrahymena thermophila: a comparison. AB - In this investigation, we compare the multiplication rates and morphogenetic responses of the two most studied Tetrahymena species, T. pyriformis and T. thermophila, at supraoptimal temperatures. Although the upper temperature limits differ greatly in the two species, the pattern of growth responses to high temperature is for the most part similar, with some differences in detail. The transient recovery of cell division at the highest temperature that allows cell division, characteristic of T. pyriformis, is observed in a less distinct form in T. thermophila. Moreover, there is a remarkable difference in developmental response, with drastic abnormalities in patterning of oral structures during the transient recovery of cell division in T. pyriformis, and far more limited abnormalities under similar conditions in T. thermophila. The abnormalities result from spatial disorder in the alignment and orientation of basal body pairs within the early oral primordium, followed by failures in the realignment that normally occurs as oral structures (membranelles and undulating membrane) mature. Both the initial spatial disorder and the failures in realignment are far more severe in T. pyriformis than in T. thermophila. PMID- 12095102 TI - An evaluation of Hsp90 as a mediator of cortical patterning in Tetrahymena. AB - This study asks two questions: 1) whether Hsp90 is involved in the regulation of cortical patterning in Tetrahymena, and 2) if it is, whether specific defects in this regulation can be attributed to functional insufficiency of the Hsp90 molecule. To address question 1, we compared the effects of a specific inhibitor of Hsp90, geldanamycin, on population growth and on development of the oral apparatus in two Tetrahymena species, T. pyriformis and T. thermophila. We observed that geldanamycin inhibits population growth in both species at very low concentrations, and that it has far more severe effects on oral patterning in T. pyriformis than in T. thermophila. These effects are parallel to those of high temperature in the same two species, and provide a tentative affirmative answer to the first question. To address question 2, we ascertained the base sequence of the genes that encode the Hsp90 molecules which are induced at high temperatures in both Tetrahymena species, as well as corresponding sequences in Paramecium tetraurelia. Extensive comparative analyses of the deduced amino acid sequences of the Hsp90 molecules of the two Tetrahymena species indicate that on the basis of what we currently know about Hsp90 both proteins are equally likely to be functional. Phylogenetic analyses of Hsp90 amino acid sequences indicate that the two Tetrahymena Hsp90 molecules have undergone a similar number of amino acid substitutions from their most recent common ancestor, with none of these corresponding to any known functionally critical region of the molecule. Thus there is no evidence that the Hsp90 molecule of T. pyriformis is functionally impaired; the flaw in the control of cortical patterning is more likely to be caused by defects in mechanism(s) that mediate the response to Hsp90, as would be expected from the "Hsp90 capacitor" model of Rutherford and Lindquist. PMID- 12095103 TI - Diversity and phylogeny of insect trypanosomatids based on small subunit rRNA genes: polyphyly of Leptomonas and Blastocrithidia. AB - With the aim of further investigating phylogenetic relationships in insect trypanosomatids, we have determined the sequences of small subunit rRNA genes from ten isolates, which were originally classified as Leptomonas, Blastocrithidia, and Wallaceina based on their morphology in the hosts. The inferred maximum likelihood, parsimony, and distance trees indicate that the Leptomonas and Blastocrithidia are polyphyletic, and confirm the polyphyly of Herpetomonas and Crithidia. Blastocrithidia triatoma and Leptomonas collosoma were among the earliest branching lineages among the insect trypanosomatids, while most other isolates were found within a closely related terminal clade, which also included Crithidia fasciculata. This analysis has clearly demonstrated that the morphological classification system of insect trypanosomatids does not always reflect their genetic affinities warranting its revision in the future. PMID- 12095104 TI - Experimental pathogenicity of a presumed monoxenous trypanosomatid isolated from humans in a murine model. AB - Two strains of a presumed lower trypanosomatid isolated from immunocompetent and HIV-infected humans in French West Indies were investigated in vitro and in vivo in a murine experimental model. The ability of parasites to grow in vitro in bone marrow-derived macrophages and their virulence in vivo were assessed. For in vivo infection, two groups of BALB/c mice were inoculated either by the subcutaneous or intravenous route with 10(7) promastigotes at day 0. Infection was monitored by measuring parasite load in liver, spleen, foot pad, popliteal, and mesenteric lymph nodes and brain from day 7 to day 150 post-infection using a microtitration technique. Parasites multiplied in mouse macrophages in vitro. In vivo, both strains proved infective to mice and capable of visceralization and dissemination in the popliteal and mesenteric lymph nodes, liver, spleen, and even brain. Both strains elicited a strong humoral response against trypanosomatid antigen in mice, which cross-reacted with Leishmania antigen. Contrasting with the straightforward dissemination of parasites, the infection was strikingly well tolerated by the murine host with no clinical signs and minimal tissue changes around parasitized macrophage infiltrates. PMID- 12095105 TI - Invasion of different cell types by sporozoites of Eimeria species and effects of monoclonal antibody 1209-C2 on invasion of cells by sporozoites of several apicomplexan parasites. AB - Sporozoites of avian Eimeria species differed markedly in their ability to invade cells in vitro. Invasion by E. tenella and E. adenoeides was significantly greater in baby hamster kidney (BHK) and chicken cecal cell (CC) cultures than in primary chicken (PCK) or turkey kidney (PTK) cell cultures. Moreover, invasion of BHK cell cultures by E. adenoeides was significantly greater than that of other Eimeria species, and invasion by E. acervulina sporozoites was significantly lower. Monoclonal antibody 1209-C2 (MAb 1209-C2) reacted by immunofluorescent labeling (IFA) with refractile bodies of sporozoites of 5 species of Eimeria and Caryospora bigenetica, but not with sporozoites of Toxoplasma gondii, Hammondia hammondi, or Cryptosporidium parvum, which have no refractile bodies. The MAb also cross-reacted with formalin-fixed BHK, CC, turkey cecal (TC) cells, and PTK. Pretreatment of BHK cells with MAb 1209-C2 significantly reduced invasion of the cells by sporozoites of E. tenella, E. acervulina, E. meleagrimitis, and C. bigenetica, but did not alter invasion by T. gondii, C. parvum, or H. hammondia. Apparently, reactivity of MAB 1209-C2 with the sporozoites was required for inhibition of invasion despite the fact that the inhibition resulted from pre treatment of the host cell. Conversely, although MAb 1209-C2 also reacted moderately with PTK and TC cells, pre-treatment of these cell cultures with the MAb did not inhibit invasion by either MAB 1209-C2-reactive or -nonreactive parasites. Collectively, the data indicated that refractile body antigens of sporozoites of Eimeria and Caryospora, which are recognized by MAb 1209-C2, may function in cellular invasion, but also suggest that cellular invasion is probably not mediated by interactions between the conserved epitopes in sporozoites and cultured host cells that are recognized by the MAb. PMID- 12095107 TI - Redescription of the rare heterotrichid ciliate, Copemetopus subsalsus Villeneuve Brachon, 1940. AB - The morphology and infraciliature of the rare heterotrichid ciliate Copemetopus subsalsus Villeneuve-Brachon, 1940 were studied in live and protargol-impregnated specimens and also by scanning electron microscopy. It is characterized by a dumbbell-shaped macronucleus, a specific dorsal brush cilia, an oblique peristome, and a distinct S-shaped adoral zone of membranelles equipped with whip shaped anterior adoral membranelles. PMID- 12095106 TI - Pneumocystis carinii f. sp. carinii synthesizes de novo four homologs of ubiquinone. AB - Ubiquinone, coenzyme Q, plays a pivotal role in electron transport and is a target for chemotherapy against a number of eukaryotic infectious agents, including Pneumocystis carinii. Coenzyme Q10 was previously identified as the major ubiquinone homolog in P. carinii isolated and purified from rat lungs; CoQ9 was also present. In contrast, CoQ9 and CoQ8 (but not CoQ10) were detected in the lungs of uninfected rat controls. These observations suggested that the pathogen synthesizes CoQ10, and perhaps CoQ9 as well. In the present study, CoQ biosynthesis in P. carinii was examined in greater detail. Radiolabeled mevalonate, a precursor of the CoQ polyprenyl chain, was incorporated in vitro into P. carinii ubiquinones. Incorporation of radiolabeled mevalonate into P. carinii CoQ was not enhanced by treating cells with lovastatin, suggesting that the cells did not transport the drug, or that a lovastatin-insensitive pathway for de novo synthesis of isoprenoids may also function in this organism. Radiolabeled precursors of the ring moiety, including shikimic acid, p hydroxybenzoic acid, and tyrosine were also incorporated into P. carinii CoQ. Unexpectedly, it was found that not only CoQ9 and CoQ10, but also CoQ7, and CoQ8, were metabolically radiolabeled by all the precursors tested, indicating that the organism synthesizes CoQ7, CoQ8, CoQ9, and CoQ10. Metabolic radiolabeling of ubiquinones in rat lung controls was not detected in experiments using either radioactive mevalonate or p-hydroxybenzoate. Thus the incorporations measured using purified P. carinii preparations were due to the enzymes of the organism. PMID- 12095108 TI - Enzymatic characterization of phospholipase D of protozoan Tetrahymena cells. AB - Phospholipase D (PLD), which is present in plant, bacterial, and mammalian cells, has been proposed to be involved in a number of cellular processes including transmembrane signaling and membrane deterioration. We demonstrated the existence of evolutionally related PLD activity in the unicellular eukaryotic protozoan Tetrahymena. The partial characterization of this enzyme showed that PLD in Tetrahymena cells was a neutral phospholipase, which catalyzed both transphosphatidylation and hydrolysis reac tions. The activity was markedly stimulated by phosphatidylinositol 4, 5-bisphosphate (PIP2) but was insensitive to phorbol 12-myristate 13-acetate (PMA) and guanosine 5'-3-O-(thio)triphosphate (GTPgammaS), suggesting that it is a PIP2-dependent PLD and that protein kinase C (PKC) and GTP-binding proteins are not implicated in the regulation of this enzyme. For its maximal activity Ca2+ was not required. This enzyme was also capable of hydrolyzing phosphatidylcholine (PC) but not phosphatidylethanolamine (PE), implying that PC was a preferred substrate. Subcellular fractionation showed that PLD-like activity localized mainly to the membrane fraction, especially microsomes. As an initial step to explore the functions of PLD in Tetrahymena, the PLD-like activity was determined during the different culture phases, and it was found to be significantly and transiently elevated in the early logarithmic phase, indicating its possible role in the development of Tetrahymena. PMID- 12095109 TI - Comparative morphology of the euglenid pellicle. II. Diversity of strip substructure. AB - The morphological diversity associated with the strip substructure of the euglenid pellicle was examined, and after identifying characters and states, we outlined hypotheses about their evolution. We have attempted to standardize terms necessary for analytical comparisons of strips by providing a glossary and comparing published synonyms. Most of the substructural diversity found in euglenids is demonstrated with 13 representative taxa. Strips are generally composed of two subcomponents: frames and projections. Frames support the basic shape of strips and many can be described as either S-shaped, plateau-shaped, M shaped, or A-shaped. Projections branch laterally from the frames, are usually periodic, and can be described as thread-like structures, an indented plate, tooth-like structures, and plate-like structures. The ancestral state included strips that were few in number, flat, and fused. The strips became S-shaped and disjoined in the lineage leading to most euglenid taxa. These strips became secondarily flattened and fused in one lineage. In some lineages of phototrophs, the strips became increasingly robust. Two strips of different morphology formed the repeating pellicular unit or doublet in four taxa. These doublets evolved convergently at least three times and may provide insights into developmental patterns of the cytoskeleton. PMID- 12095110 TI - The Euplotes crassus conjugation-specific conN1 gene encodes a transcription elongation factor TFIIS-like protein. AB - The Euplotes crassus macronuclear DNA molecule containing the conjugation specific conN1 gene has been sequenced, along with a cDNA clone. The results indicate that the conN1 gene encodes a protein similar to the transcription elongation factor TFIIS proteins identified in other eukaryotes. PMID- 12095111 TI - Organization of telomeres during the cell and life cycles of Trypanosoma brucei. AB - The genome of Trypanosoma brucei contains about 120 chromosomes, which do not visibly condense during mitosis. We have analyzed the organization and segregation of these chromosomes by in situ hybridization using fluorescent telomere probes. At the onset of mitosis, telomeres migrate from their nuclear peripheral location and congregate into a central zone. This dense group of telomeres then splits into two entities that migrate to opposite nuclear poles. Segregation continues until the double-sized nucleus divides and, before cytokinesis occurs, the telomeres reorganize into the discrete foci observed at interphase. During migration, the telomeres are located at the free end of the mitotic spindle. Treatment with the microtubule polymerization inhibitor rhizoxin prevents telomere clustering and chromosomal segregation. In the insect-specific procyclic form as well as in the non-dividing bloodstream stumpy form, telomeres tend to cluster close to the nuclear periphery at interphase. In contrast, in the proliferative bloodstream slender form the telomeres preferentially locate in the central zone of the nucleus. Thus, telomeres are closer to the nuclear periphery during those life cycle stages where the telomeric expression sites for the variant surface glycoprotein are all inactive, suggesting that transcriptional inactivation of these sites is related to their subnuclear localization. PMID- 12095112 TI - Pseudoloma neurophilia n. g., n. sp., a new microsporidium from the central nervous system of the zebrafish (Danio rerio). AB - An unusual xenoma-forming microsporidium was discovered in the central nervous system of moribund zebrafish from a laboratory colony in Eugene, Oregon. Infected fish were often emaciated and lethargic, and histological examination commonly revealed severe myelitis and myositis associated with the infection. Based on its structure, development, and small subunit ribosomal DNA sequence it is unique among fish microsporidia. Spores are uninucleate, ovoid to pyriform, with a prominent posterior vacuole. Spores average 5.4 x 2.7 microm with 13-16 coils of the polar filament. The microsporidium produces xenomas within the spinal cord and hindbrain of fish, and xenomas contained sporophorous vesicles with up to 16 spores. Sporoblasts and presporoblast stages (probably sporonts) are found occasionally in small aggregates dispersed randomly throughout xenomas. It clustered in the "Ichthyosporidium group" along with other fish microsporidian genera based on rDNA sequence analysis. The rDNA sequence of the zebrafish microsporidium was most similar to that of Ichthyosporidium, but showed only 12.1% similarity and therefore this microsporidium can be considered a distinct genus and species, which we have named Pseudoloma neurophilia n. g., n. sp. PMID- 12095113 TI - Fine oral filaments in Paramecium: a biochemical and immunological analysis. AB - In Paramecium, several kinds of the oral networks of fine filaments are defined at the ultrastructural level. Using the sodium chloride-treated oral apparatus of Paramecium as an antigen to produce monoclonal antibodies, we have begun to identify the proteins constituting these networks. Immunoblotting showed that all positive antibodies were directed against three bands (70-, 75-and 83-kD), which corresponded to quantitatively minor components of the antigen; there was no antibody specific for the quantitatively major components (58- and 62-kD). Immunolocalization with four of these antibodies directed against one or several of these three bands showed that these proteins are components of the fine filaments supporting the oral area; a decoration of the basal bodies and the outer lattice was also observed on the cortex. Immunofluorescence on interphase cells suggested that the three proteins colocalized on the left side of the oral apparatus, whereas only the 70-kD band was detected on the right side. During division, the antigens of the antibodies were detected at different stages after oral basal body assembly. The antibodies cross-reacted with the tetrins, which are oral filament-forming proteins in Tetrahymena, demonstrating that tetrin related proteins are quantitatively minor components of the oral and the somatic cytoskeleton of Paramecium. PMID- 12095114 TI - Balance and role playing. PMID- 12095115 TI - Failure of hallux MP preservation surgery for rheumatoid arthritis. AB - Eight patients underwent surgery on 15 feet for rheumatoid forefoot problems. Thirteen of the 15 feet that were operated upon had an attempt to preserve the hallux metatarsophalangeal joint while resectional arthroplasty was performed on the lesser MP joints. All of the 13 feet that had the MP joint preserved had a well-preserved joint space preoperatively and no active signs of inflammation at the time of this procedure. Eight feet underwent a distal Chevron osteotomy to realign the great toe, two feet underwent an IP fusion as only the IP joint had evidence of erosive changes, and one foot underwent a combination of a Chevron osteotomy and a proximal phalangeal osteotomy (Akin procedure). Two patients had no surgery on their first ray as it was well aligned with no evidence of erosive changes. Of the 13 feet that did not have a fusion performed, 11 had development of a valgus deformity or inflammatory erosions. The average time to failure was 24 months (range, six to 36 months). The Chevron/Akin procedure remained successful at 18 months and one of the IP fusions was successful at six years after surgery. Although patients with rheumatoid forefoot disease may on occasion have a well-preserved hallux MP joint with minimal or no deformity and no active inflammation, with severe lesser toe involvement, most of these patients will fail a surgical procedure which does not involve fusion of the hallux MP joint. PMID- 12095116 TI - The arterial supply of the lesser metatarsal heads: a vascular injection study in human cadavers. AB - The possibility of avascular changes of the metatarsal heads following forefoot surgery has been previously documented. The aim of this study was to investigate the arterial supply of the lesser metatarsal heads with regard to osteotomies of these bones. We used epoxy resin injections and a modified Spalteholz technique in human cadaveric specimens to demonstrate the intraosseous and extraosseous blood supply of the lesser metatarsals. The metatarsal heads had two arterial sources: 1. The dorsal metatarsal arteries, which arose from the dorsalis pedis artery, and 2. The plantar metatarsal arteries, which are branches of the posterior tibial artery. These two vessels typically anastomosed at two sites about the metatarsal heads, forming a vascular ring and provided an extensive extraosseous arterial network around the metatarsal heads. Small arterial branches of this network run distally on the metatarsal cortex to enter the bone of the metatarsal head. The nutrient arteries traversed the cortex of the metaphysis close to the capsular and ligamentous insertions to provide multiple branches for the supply of the subchondral bone. Extensive capsular stripping during metatarsal head osteotomies results in damage to the medial and lateral head vessels. PMID- 12095117 TI - First metatarsophalangeal arthrodesis: a clinical, pedobarographic and gait analysis study. AB - This study investigated the results of first metatarsophalangeal (MTP) arthrodesis in terms of clinical outcome measures, plantar pressure distribution, and gait patterns. Ten feet in nine patients with severe hallux rigidus (HR) who underwent first MTP arthrodesis were studied. The preoperative evaluation included a subjective questionnaire, physical exam, AOFAS hallux score, radiographs and dynamic pedobarography (EMED). At follow-up (average 34 months) these were repeated, and gait analysis studies were obtained. Patients showed significant clinical improvement based on the subjective criteria. The mean AOFAS score improved from 38 preoperatively to 90 postoperatively. Postoperative EMED analysis showed restoration of the weightbearing function of the first ray, with greater maximum force carried by the distal hallux at toe-off. Kinematic and kinetic gait analysis from each patient's operative limb were compared to the unaffected contralateral limb and to age- and sex-matched healthy subjects. The kinematic data indicated a significantly shorter step length with some loss in ankle plantar flexion at toe-off on the fused side. The kinetic data indicated a reduction in both ankle torque and ankle power at push-off. Clinical results indicated effective pain relief and a high level of patient satisfaction, consistent with previous reports in patients with symptomatic Hallux Rigidus. PMID- 12095118 TI - Chevron (Austin) distal metatarsal osteotomy for hallux valgus: comparison of pre and post-surgical characteristics. AB - The purpose of the study was to compare the range of motion, perceived pain and plantar loading characteristics of the Chevron (Austin) corrective procedures in treating hallux valgus (HV) 12 months postsurgically. Twenty-five female participants with the diagnosis of mild to moderate HV deformity were studied. All participants had a distal metatarsal osteotomy (Chevron (Austin)) to correct their deformity. First metatarsophalangeal and talocrural joint range of motion (ROM) and a 10-point analog pain scale were measured presurgically and 12 months post-surgically on each participant. Radiographic measures of hallux valgus and intermetatarsal (IM) angles were taken preoperatively and six weeks postoperatively for comparison. Five pressure distribution measurements were recorded of barefoot walking using the EMED-SF presurgically and 12 months postsurgically. Statistical analyses revealed that plantar loading is still altered 12 months postsurgically despite a decrease in perceived pain and adequate first metatarsophalangeal joint ROM. Greater loading occurred in the central forefoot (CFF) region with decreases in some of the loading parameters in the medial toe (MT) region postsurgically. Loading parameters in the lateral forefoot (LFF), heel (HL), midfoot (MF), and lateral toe (LT) were unchanged 12 months postsurgery compared to presurgery. PMID- 12095119 TI - Benefits of early prosthetic management of transtibial amputees: a prospective clinical study of a prefabricated prosthesis. AB - To evaluate the use of an immediate postoperative prosthesis (IPOP) for transtibial amputees, we compared patient outcomes from a prospective clinical study of 19 patients managed with an IPOP with those of a retrospective review of a matched historic control group of 23 patients managed with standard soft dressings. Data were analyzed with the Student's t-test, and significance was set at P = 0.05. The IPOP patients had no surgical revisions, whereas the patients with standard soft dressings had 11. This was a significant difference. IPOP patients also had significantly fewer postoperative complications and shorter times to custom prosthesis than did controls. PMID- 12095120 TI - Elongation behavior of calcaneofibular and cervical ligaments in a closed kinetic chain: pathomechanics of lateral hindfoot instability. AB - Numerous reconstructive procedures are performed to correct both ankle and subtalar instability after trauma although the precise pathology which results in this chronic instability and pain is not yet known. This study examined the role of the calcaneofibular (CLFL) and cervical ligaments (CRVL) during physiologic loading and demonstrated the effect of CLFL deficiency on the CRVL. Talar and subtalar tilt as well as inversion range of motion before and after CLFL sectioning were studied. Eleven osteoligamentous fresh frozen cadaver legs were used in which each foot was taken through six positions: neutral, 35 degrees plantarflexion, dorsiflexion, inversion, plantarflexion-inversion, and dorsiflexion-inversion. The CLFL and CRVL stretched the greatest in dorsiflexion inversion. The most interesting finding was that the CRVL was elongated relative to neutral in all other test positions of the foot. However, the CLFL was shortened relative to neutral in plantarflexion and plantarflexion-inversion. In the CLFL deficient state, CRVL ratios demonstrated significant increases in length of the CRVL. Talar tilt increased on average more than 9 degrees with CLFL deficiency (p < 0.008) while subtalar tilt did not change significantly. The maximum tibiocalcaneal angle, recorded for dorsiflexion-inversion, increased more than 5 degrees after sectioning the CLFL (p < 0.05). PMID- 12095121 TI - Osteolysis of structural autograft after calcaneocuboid distraction arthrodesis for stage II posterior tibial tendon dysfunction. AB - Surgical treatment of stage II posterior tibial tendon dysfunction that fails nonoperative treatment is amenable to operative treatment. This commonly consists of a medial soft-tissue reconstruction and lateral column lengthening. We report on 32 patients undergoing distraction calcaneocuboid arthrodesis using an autogenous tricortical iliac crest graft. Almost 50% of patients did not go on to complete and uneventful incorporation of the graft at the arthrodesis site. Two different types of failure were noted. The first was classic nonunion which maintained structural integrity of the graft. The second was osteolysis and collapse of the graft accompanying the nonunion. Risk factors included smoking, K wire fixation and possibly larger deformities requiring Achilles tendon lengthening. Complications of nonunion were salvageable with reoperation. PMID- 12095122 TI - Distal tarsal tunnel release with partial plantar fasciotomy for chronic heel pain: an outcome analysis. AB - This study evaluated the effectiveness of distal tarsal tunnel release with a partial plantar fasciotomy for chronic subcalcaneal pain syndrome in patients who failed nonoperative management through a retrospective review of all patients undergoing this procedure between 1994 and 1999. Distal tarsal tunnel release and a partial plantar fasciotomy were offered only to those patients with disabling heel pain and were performed under general anesthesia or ankle block. Seventy five patients (80 heels), averaging 20 months of nonoperative treatment, were identified (group I). Postoperative outcome questionnaires (SF-36 and Foot Function Index [FFI]) were sent to these patients and 44 (46 heels) responded (group II). In group I, 88% of patients had a good to excellent result. Many continued with mild to moderate residual symptoms, which typically did not limit their activity. In group II, 91% of patients were somewhat to very satisfied with their outcome. Visual analogue scale scores for pain were reduced by a mean of 55. SF-36 scores, matched against a control group of patients receiving just nonoperative treatment, showed a statistically significant improvement in all pain and functioning subcategories. We conclude that a distal tarsal tunnel release with a partial plantar fasciotomy may successfully increase function and decrease pain in patients who fail nonoperative treatment. PMID- 12095123 TI - An evaluation of two foot-specific, health-related quality-of-life measuring instruments. AB - In the past decade, health-related quality-of-life (HRQoL) assessment has become an important tool to measure change as a result of treatment in clinical trials. The aim of this project was to compare and contrast two foot-specific HRQoL questionnaires: the Foot Function Index (FFI) and the Foot Health Status Questionnaire (FHSQ). To assist in this process, a quasi-experimental trial was performed to evaluate the effectiveness of foot orthoses in improving HRQoL in people suffering from plantar fasciitis. The results from this study found that the FFI is generally less responsive to change, particularly in the domain of Activity Limitation. Nevertheless, significant improvements were found for two out of the three domains (Pain and Disability, but not Activity Limitation) for the FFI, as well as the overall score of FFI. In contrast, all four domains (Pain, Function, Footwear and General Foot Health) of the FHSQ showed significant improvement. Our experience with the questionnaires also revealed that certain questions in the Activity Limitation domain of the FFI can lead to inconsistent scoring, thus casting doubts over the suitability of this domain for use with patients with plantar fasciitis. Therefore, the FHSQ has several advantages when evaluating HRQoL in patients being treated with foot orthoses for plantar fasciitis, and should be viewed as the preferred questionnaire. PMID- 12095124 TI - Anatomical study of the medial ankle ligament complex. AB - The purpose of this in vitro study was to determine insertion area, length and thickness of the various bundles and their anatomical relationship with inter individual differences. Twelve ankles from human cadavers (ages 56 to 95 years, from nine men and three women) were dissected to the capsuloligamentous structures. Marked inter-individual differences were found for the five main ligaments (tibiospring, tibiocalcaneal, posterior and anterior deep tibiotalar and superficial posterior tibiotalar). The tibionavicular ligament is a thickened fibrous layer of the ankle capsule. The tibiocalcaneal and tibiospring ligaments are the longest, and the tibiocalcaneal and posterior deep tibiotalar ligaments are the thickest of these ligaments. Fibrils run in the direction of the tibia or dorsally. Knowledge of the deltoid ligament complex is necessary for anatomically and biomechanically correct reconstruction that provides stability without hazard to biomechanics of function. PMID- 12095125 TI - Visualization of ankle tendons and ligaments with MR imaging: influence of passive positioning. AB - PURPOSE: To determine if passive positioning of the ankle influences the MR imaging appearance of ankle tendons and ligaments. METHODS: A positioning device was used during imaging of 10 volunteers. Axial and coronal T1-weighted images were acquired in six positions of the ankle, and the appearance of the tendons and lateral ligaments was subjectively evaluated. RESULTS: The peroneus longus (PL), peroneus brevis (PB), anterior tibialis (AT), extensor digitorum longus (EDL), and extensor hallucis longus (EHL) tendons were best visualized in 20 degrees of plantarflexion and 20 degrees of inversion (p < 0.0001). The anterior talofibular (ATF) and calcaneofibular (CF) ligaments were best seen in 20 degrees of plantarflexion (p < 0.0001). Ten degrees of dorsiflexion was the least useful imaging plane for all of the aforementioned structures. CONCLUSION: When there is clinical concern regarding pathology of a specific structure about the ankle, passive positioning may allow optimal evaluation. PMID- 12095126 TI - Effect of dopamine-induced changes in splanchnic blood flow on MEGX production from lidocaine in septic and cardiac surgery patients. AB - The production of monoethylglycine xylidide (MEGX) from lidocaine is thought to be dependent on hepatic blood flow. We assessed the relationship between hepatosplanchnic blood flow, lidocaine uptake, and the production of MEGX from lidocaine in seven patients after cardiac surgery and in nine septic patients. Systemic (pulmonary artery catheter) and splanchnic (hepatic vein catheter and dye dilution) hemodynamics and arterial and hepatic venous lidocaine and MEGX concentrations were measured after a lidocaine bolus injection (1 mg/kg) before and 90 min after increasing cardiac output by at least 25% with dopamine. Dopamine infusion [in cardiac surgery patients 4.2 (1.4-8.5) microg x kg(-1) x min(-1) (median, range) and in septic patients 4.0 (2.1-9.0) microg x kg(-1) x min(-1)] increased splanchnic blood flow in cardiac surgery patients from 0.65 (0.12) L x min(-1) x m(-2) to 0.84 (0.14) L x min(-1) x m(-2) mean (standard deviation) P = 0.018) and in septic patients from 0.91 (0.32) L x min(-1) x m(-2) to 1.12 (0.40) L x min(-1) x m(-2) (P = 0.038). Splanchnic MEGX production for the 30 min after lidocaine injection was higher in cardiac surgery patients than in septic patients both at baseline [4130 (1100) microg x m(-2) vs. 930 (420) microg x m(-2) (P < 0.005)] and afterdopamine infusion [4480 (1000) microg x m( 2) vs. 1090 (620) microg x m(-2) (P = 0.005)]. We found no correlation between changes in MEGX production and changes in splanchnic blood flow. Patients with sepsis have severe impairment of cytochrome P450-dependent liver function, which is not influenced by acute changes in hepatosplanchnic blood flow. MEGX production cannot be used as an estimate of changes in splanchnic blood flow. PMID- 12095127 TI - Bioactive oxidized lipids in the plasma of cardiac surgical intensive care patients. AB - Critical illness is associated with increased oxidative stress that may give rise to the formation of lipid hydroperoxides (LOOH) and various secondary degradation products such as fragmented phosphatidylcholine (FPC) and lipids related to the platelet-activating factor (PAF). Because some oxidized phospholipids are potent proinflammatory agents, we measured the concentration of LOOH, FPC, and PAF-like activity in blood plasma of 36 patients who had undergone cardiac surgery and developed postoperative complications associated with systemic inflammatory response syndrome (SIRS) or multiple organ failure (MOF). These patients were compared to two control groups, namely preoperative patients scheduled for cardiac surgery (n = 13), and postoperative patients without complications (n = 19). Postoperative patents had higher concentrations of LOOH and lower concentrations of FPC than preoperative patients (P < 0.01). However, SIRS and MOF had no significant effect on the concentration of oxidatively modified lipids. This is despite the fact that MOF patients showed evidence of increased lipid peroxidation (7-fold higher ratio of alpha-tocoquinone/alpha-tocopherol compared to control). LOOH correlated positively with the white blood cell count. Postoperative patients had 4-fold higher plasma activities of phospholipase A2 and this activity was further increased in patients with SIRS (P < 0.04). Phospholipase A2 activity correlated negatively with the concentration of FPC. The data suggest that oxidatively modified lipids do not accumulate in patients with SIRS and MOF, perhaps because enhanced peroxidation of lipids is offset by enhanced lipolytic activity. PMID- 12095128 TI - Attenuated monocyte IL-10 production in glucose-6-phosphate dehydrogenase deficient trauma patients. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of the world's most common known human genetic polymorphisms, but the pathophysiology of the defect remains largely unknown. In the present study, we compared hematology parameters and ex vivo monocyte cytokine responses in non-deficient and G6PD-deficient trauma patients. Deficient and non-deficient, moderately injured, trauma patients exhibited similar hematology profiles at the time of hospital admission. In contrast to non-deficient patients, G6PD-deficient patients were anemic 2 days post-injury. Monocytes from deficient individuals produced 50% less interleukin 10 (IL-10) in response to LPS and >90% less IL-10 in response to PMA, compared with non-deficient patients, 2 days post-injury. The presence of phenylhydrazine treated, opsonized, autologous RBC (OX-RBC), alone had no effect on IL-10 production by non-deficient or deficient monocytes, whereas IL-10 responses to lipopolysaccharide (LPS) were augmented by OX-RBC in both groups. However, IL-10 production was markedly lower by monocytes from G6PD-deficient than non-deficient patients after stimulation with LPS plus OX-RBC. TNF-alpha production following PMA was similar in deficient and non-deficient patients, and the differences following LPS or LPS plus OX-RBC stimulation were moderate between deficient and non-deficient samples. Interferon (IFN)-gamma production ex vivo was doubled by OX-RBC treatment alone, but it was not stimulated by LPS treatment. IFN-gamma production was similar in non-deficient and G6PD-deficient patients. These data suggest that the observed differences in IL-10 responses between G6PD-deficient and non-deficient patients are not attributable to differences in TNF-alpha or IFN-gamma production. Taken together, our data suggest that a reduction in the capacity to produce IL-10 may be an intrinsic characteristic of G6PD-deficient monocytes. An attenuated IL-10 production may be a contributing mechanism in the previously observed augmented inflammatory response in severely injured G6PD deficient compared with non-deficient trauma patients. PMID- 12095129 TI - Early interleukin-10 treatment improves survival and enhances immune function only in males after hemorrhage and subsequent sepsis. AB - Recent studies have demonstrated gender differences in the immune response following hemorrhagic shock with an enhanced immune function and lower mortality following subsequent sepsis in females. Early interleukin-10 (IL-10) treatment has been shown to have beneficial effects on the depressed immune function in males, but not in females following shock. However, it remains unclear if the observed gender-related effect of IL-10 treatment results in an advantage following subsequent polymicrobial sepsis. To study this, male and female CBA/J mice (age 2-3 months) were subjected to hemorrhage (35 +/- 5 mmHg for 90 min and fluid resuscitation). At resuscitation, each received either 10 microg of recombinant murine IL-10 or placebo i.p.. At 48 h after resuscitation, either peritoneal macrophages (pMphi) and plasma were harvested, or polymicrobial sepsis was induced by cecal ligation and puncture (CLP). Following CLP, either survival over 10 days was measured, or pMphi and plasma were harvested 4 h after CLP to assess TNF-alpha, IL-6, IL-10, and prostaglandin E2 (PGE2) release of pMphi and plasma levels of IL-10, free testosteron, and 17-beta estradiol. Early IL-10 treatment restored depressed proinflammatory immune response in males (TNF-alpha and PGE2), which was associated with an enhanced survival (P < 0.05) following subsequent sepsis as compared with placebo-treated mice (8/20 and 1/20, respectively). In contrast, the immune response and survival in females receiving IL-10 was not significantly changed, although females treated with IL-10 had a trend towards higher mortality (7/15 and 2/15, respectively; P = 0.08). Thus, early IL-10 anti-inflammatory treatment following hemorrhage has potential beneficial effects only in males associated with enhanced survival following subsequent sepsis. PMID- 12095130 TI - Hyperosmolarity abrogates neutrophil cytotoxicity provoked by post-shock mesenteric lymph. AB - Hypertonic saline (HTS) resuscitation inhibits acute lung injury in animal models of shock, but some argue this may simply represent more efficient fluid resuscitation. Inflammatory mediators within mesenteric lymph have been identified as a link between splanchnic hypoperfusion and acute respiratory distress syndrome (ARDS). We hypothesize that HTS resuscitation abrogates post shock lymph-mediated neutrophil (PMN) priming and PMN-mediated human endothelial cell cytotoxicity. Mesenteric lymph was collected from rats (n = 5) before (control), during non-lethal hemorrhagic shock, defined as a mean arterial pressure (MAP) of 40 mmHg for 30 min, and after resuscitation (shed blood + 2 x lactated Ringers (LR) versus 7.5% NaCl, 4 cc/kg, over 5 min). Isolated human PMNs were primed with physiologic concentrations (5% v:v) of lymph either from animals resuscitated with LR or HTS and activated with either PMA or fMLP. In a separate set of experiments, human PMNs were primed with LR lymph after incubation with HTS (180 mM NaCl). The maximal rate of superoxide production was measured by reduction of cytochrome C. In addition, the effect of HTS pretreatment on PMN adherence to human pulmonary microvascular endothelial cells (HMVEC) and PMN mediated cytotoxicity was determined after lymph-mediated PMN priming. PHSML primed isolated PMNs above buffer controls and pre-shock lymph in a normotonic environment; HTS resuscitation abrogated this effect. HTS preincubation of isolated PMNs inhibited PHSML-induced PMN priming, adherence to HMVECs, and PMN mediated HMVEC cytotoxicity. Hypertonic resuscitation (HTS) abrogates PHSML pniming of the PMN and PMN-mediated HMVEC cytotoxicity. Furthermore, incubation of PMNs in clinically relevant HTS (180 mM NaCl) prevents PHSML PMN priming and PMN:HMVEC interactions. These studies suggest inhibition of PMN signal transduction is a mechanism whereby HTS resuscitation abrogates acute lung injury. PMID- 12095131 TI - The stress response decreases NF-kappaB activation in liver of endotoxemic mice. AB - Recent studies suggest that the stress (heat shock) response protects cells and tissues from inflammatory and other noxious insults. The transcription factor nuclear factor-kappa B (NF-kappaB), normally sequestered in the cytoplasm by its inhibitory protein IkappaB, regulates many genes involved in the inflammatory response to critical illness. Endotoxemia is associated with increased NF-kappaB activity in liver but the effect of the stress response on endotoxin-induced NF kappaB activation in the liver is not known. We hypothesized that the stress response inhibits NF-kappaB DNA binding activity in liver during endotoxemia. The stress response was induced in mice by hyperthermia (42 degrees C for 3 min) or sodium arsenite (10 mg/kg) and resulted in increased hepatic heat shock protein 72 levels. After induction of the stress response, mice were injected subcutaneously with endotoxin (12.5 mg/kg) or a corresponding volume of sterile saline. NF-kappaB DNA binding activity in the nuclear fraction of liver tissue increased and cytoplasmic IkappaB-alpha levels decreased after endotoxin injection, with a maximal effect seen at 60 min. The endotoxin-induced increase in NF-kappaB DNA binding activity and decrease in IkappaB-alpha levels were inhibited by prior induction of the stress response. In additional experiments, treatment of mice with sodium arsenite after induction of endotoxemia blunted the increase in NF-kappaB activity, indicating a therapeutic potential of sodium arsenite, in addition to its preventive effect. The present results suggest that the protective effects of the stress response in vivo may, at least in part, be due to inhibited NF-kappaB activation. PMID- 12095132 TI - Activation of stress-responsive pathways by the sympathetic nervous system in burn trauma. AB - We have shown previously that bum trauma activates the stress responsive proteins, p38 mitogen-activated protein kinase (MAPK), c-jun NH2-terminal kinase (JNK), and NF-kappaB, and we have shown further that p38 MAPK is an important mediator of cardiomyocyte TNF-alpha secretion and cardiac dysfunction in burn trauma. Since burn trauma causes a rise in circulating catecholamine levels, we hypothesized that this increased sympathetic activity may function as an upstream activator of the p38 MARK pathway in burn trauma. This study determined whether the alpha1-adrenergic receptor ligand phenylephrine could mimic burn trauma activation of p38 MAPK, JNK, and NF-kappaB nuclear translocation; and the effect of the alpha1-adrenergic receptor antagonist prazosin on either phenylephrine or burn-mediated activation of the stress response pathway was examined. Sprague Dawley rats were divided into seven groups: Group 1, controls; Group 2, phenylephrine-treated (2 microg/kg, i.v.) control rats; Group 3, phenylephrine treated plus prazosin-treated (1 mg/kg, i.v.) control rats; additional rats were given burn over 40% total body surface area (TBSA) and received vehicle (1 mL of 2% sucrose, p.o.) plus fluid resuscitation (Group 4), while in Group 5, burn rats were given prazosin (1 mg/kg, p.o.) plus fluid resuscitation. In Groups 6 and 7, sham-burned rats were given either vehicle (1 mL of 2% sucrose, p.o.) or prazosin (1 mg/kg, p.o.) to provide appropriate controls. Administration of phenylephrine to rats caused a significant activation of cardiac p38 MAPK/JNK activities (Western blot) and cardiac NF-kappaB nuclear translocation (electrophoretic mobility shift assay, EMSA). Prazosin blocked phenylephrine mediated changes in p38 MAPK/JNK activities. Burn trauma activated cardiac p38 MAPK/JNK and NF kappaB, increased TNF-alpha secretion by cardiomyocytes, and impaired cardiac function. Prazosin treatment in burns interrupted the burn-mediated signaling cascade, decreasing TNF-alpha secretion by cardiomyocytes and preventing post burn cardiac contractile dysfunction. Thus, burn trauma-related sympathetic activity likely activates the stress-responsive cascade, which regulates myocardial TNF-alpha transcription/translation and culminates in cardiac contraction and relaxation defects. PMID- 12095133 TI - Epinephrine as a mediator of pulmonary neutrophil sequestration. AB - Neutrophil-mediated lung injury is a potential complication of trauma and sepsis. Concomitant with trauma and sepsis, there is an immediate and sustained systemic elevation of catecholamines including epinephrine. In the absence of trauma or sepsis, we examined whether epinephrine contributes to the accumulation of neutrophils in the lung. Eight- to 12-week-old male CF-1 mice were injected i.p. with 0.2 mL of normal saline or epinephrine (0.1-5.0 mg/kg). An unmanipulated control group was included to examine the stress of i.p. injection. Animals were sacrificed at predetermined time points, and lung and spleen were harvested. PMN accumulation was assessed by using a myeloperoxidase (MPO) assay, which is an indirect marker for neutrophil presence. Morphometric analysis of lung tissue was performed by a pathologist blinded to the groups. Increasing epinephrine doses resulted in a significantly increased accumulation of pulmonary neutrophils compared with normal saline. The stress of normal saline injection also resulted in a significantly greater pulmonary neutrophil accumulation than unmanipulated controls. The effects of epinephrine on pulmonary neutrophil accumulation were greatest at 2 h, but they were not significantly different from saline-injected controls by 12 h. These results correlated with histological analysis. There were no significant differences in spleen MPO activity between groups, suggesting an organ-specific mechanism of epinephrine-induced pulmonary neutrophil sequestration. In the absence of trauma, shock, or infection, epinephrine results in the accumulation of neutrophils in murine lungs. The finding that "injection stress" increased lung neutrophil sequestration suggests the possibility that this mechanism may be physiologically relevant. Thus, epinephrine release in trauma may set the stage for development of neutrophil-mediated acute lung injury. PMID- 12095134 TI - Monocyte adhesion and transmigration induce tissue factor expression: role of the mitogen-activated protein kinases. AB - The expression of tissue factor (TF) by monocytes that have transmigrated across the endothelium to sites of extravascular inflammation acts both to focus and amplify the inflammatory response. Because clustering of the integrins responsible for endothelial adhesion and transmigration induces tyrosine phosphorylation and activation of the mitogen-activated protein (MAP) kinases, we postulated that transmigration might lead to monocyte activation and TF production. Monocytes were migrated across TNFalpha-primed ECV304 cells grown on fibronectin-coated Transwell chambers in response to FMLP (10(-8) M). After transmigration, monocytes showed a time-dependent increase in surface TF expression and biological procoagulant activity. TF expression was dependent on monocyte adhesion to ECV304 cells. Specifically, TF was not induced by FMLP treatment of suspended monocytes, by migration across fibronectin alone, or by soluble factors induced during migration, whereas monocyte-ECV304 adhesion was sufficient to stimulate TF. Antibodies against CD29 (beta1 integrin), but not against CD18 (beta2 integrin) or CD31 (PECAM-1), inhibited TF expression. Monocyte adhesion to ECV304 cells induced tyrosine phosphorylation of cellular proteins and specifically of the ERK and p38 MAP kinases. Tyrosine kinase inhibition with genistein (10 microg/mL) blocked transmigration, whereas selective ERK inhibition with PD98059 (50 microM) or p38 inhibition with SB203580 (20 microM) did not. However, both ERK and p38 inhibition dose dependently abolished TF expression. These studies suggest that an extravascular focus of infection or inflammation can promote both intravascular thrombosis and extravascular fibrin deposition during the process of adhesion and transmigration across the endothelial barrier. The selective inhibition of the mitogen-activated protein kinases may offer a novel therapeutic means of modulating this inflammatory sequence. PMID- 12095135 TI - Intracellular antioxidant activity is necessary to modulate the macrophage response to endotoxin. AB - The tissue-fixed macrophage (Mphi) is a key cell in the coordination of the excessive systemic immunoinflammatory response underlying the adult respiratory distress syndrome (ARDS). Macrophage-generated reactive oxygen intermediates (ROIs) are involved in both tissue destruction via lipid peroxidation and in the activation of these inflammatory cells. It is unclear whether oxidant-induced activation involves an extracellular effect and membrane destabilization or occurs through intracellular alteration of the redox state and direct involvement as second messengers. In this study, we compare the differential effects of known intracellular vs. extracellular antioxidants on the Mphi response to endotoxin. Rabbit alveolar Mphi were obtained by bronchoalveolar lavage and exposed to either the extracellular antioxidants [vitamin C (VC) (10-1000 microM), Trolox (100-1000 microM, superoxide dismutase (SOD) (10-500 microM))] or the intracellular antioxidants [N-acetylcysteine (NAC) (0.1-10 mM) or butylated hydroxyanisole (BHA) (10-200 microM)] for 1 h. Cells were subsequently stimulated with lipopolysaccharide at 10 ng/mL. After 18 h, supernatants were analyzed for tumor necrosis factor (TNF) and F2 isoprostane (F2ISP) production and cellular monolayers for procoagulant activity (PCA). A dose response inhibition of both TNF and PCA production was demonstrated after both NAC and BHA pretreatment but not with VC, Trolox, or SOD. In addition, northern blots revealed inhibition of TNF mRNA production by both NAC and BHA. F2ISP, a marker of membrane lipid peroxidation, was inhibited by BHA and Trolox but not NAC, VC, or SOD. In conclusion, antioxidants that are incorporated intracellularly are expected to be beneficial in the treatment of excessive inflammatory responses through the interruption of redox dependent signal transduction pathways and subsequent modulation of the Mphi proinflammatory response. PMID- 12095136 TI - Carboxy-PTIO, a scavenger of nitric oxide, selectively inhibits the increase in medullary perfusion and improves renal function in endotoxemia. AB - The acute renal failure associated with septic shock is associated with a high mortality despite dialytic therapies. Endotoxemia leads to marked changes in the distribution of intrarenal perfusion that may be independent of alterations in total renal blood flow or systemic hemodynamics. Modulation of this intrarenal redistribution may protect against acute renal failure. This study examines the effect of carboxy-2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (carboxy-PTIO), a scavenger of nitric oxide (NO), on systemic and intrarenal hemodynamics measured by laser Doppler flowmetry following the induction of endotoxemia in the anesthetized rat. Infusion of lipopolysaccharide (LPS) led to a prompt reduction in inulin clearance at 60 min, which remained reduced for 6 h in saline-treated rats. Administration of carboxy-PTIO led to a sustained increase in inulin clearance over 360 min post-LPS. During endotoxemia, cortical perfusion fell acutely by 29 +/- 8%, whereas medullary perfusion increased by 71 +/- 11%. The increase in medullary perfusion was potently and selectively inhibited by carboxy-PTIO. We propose that inhibition of medullary hyperemia maintains glomerular hydrostatic pressure, thus leading to the improved renal function during endotoxemia and that scavenging of NO may prove to be a useful therapeutic option in the acute renal failure associated with septic shock. PMID- 12095137 TI - Differences in neutrophil death among beta-lactam antibiotics after in vitro killing of bacteria. AB - Antibiotic therapy is an essential treatment for gram-negative bacterial infections. Antibiotic-induced endotoxin release and subsequent production of inflammatory cytokines reportedly depend on the type of antibiotic action. This study examined the effects of various beta-lactam antibiotics on cell death of human polymorphonuclear neutrophils (PMNs) cocultured with Escherichia coli (E. coli) in vitro. E. coli morphology after antibiotic treatment was determined. PMNs and E. coli were cocultured with antibiotics for 0, 4, or 12 h. Levels of endotoxin and cytokines (TNF-alpha, IL-1beta, and IL-6) in the supernatants were measured. The filtrates of antibiotic-treated E. coli supernatants were cocultured with PMNs for 0, 4, or 12 h. In all experiments, ampicillin (ABPC), cefazolin sodium (CEZ), cefoperazone sodium (CPZ), latamoxef sodium (LMOX), imipenem (IPM), and polymyxin B sulfate (PLB) were used at 30 microg/mL. PMNs were isolated from healthy volunteers. PMN cell death was assessed by flow cytometry and light microscopy. ABPC, CEZ, CPZ, and LMOX, which induce bacterial filament formation with lysis, caused PMN necrosis when cocultured with E. coli. In contrast, IPM, which induces bacterial spheroplast formation with lysis, caused PMN apoptosis. Levels of endotoxin, TNF-alpha and IL-6 in the supernatants with IPM and PLB were significantly lower than in those with other beta-lactam antibiotics. The filtrates of IPM- and PLB-treated E. coli supernatants induced PMN apoptosis, whereas those treated with other beta-lactam antibiotics increased PMN necrosis. Beta-lactam antibiotics have different impacts on the types of PMN cell death after E. coli killing. Underlying mechanisms and the clinical relevance of IPM-induced PMN apoptosis in severe gram-negative infection warrant further investigation. PMID- 12095138 TI - The effect of tumor necrosis factor-alpha on microvascular permeability in an isolated, perfused lung. AB - This study examines the hypotheses that TNF-alpha causes a dose-dependent increase in the microvascular permeability of ex vivo buffer perfused lungs that is quantitatively similar to that caused by lipopolysaccharide (LPS) or thromboxane A2 (TxA2). We also postulated that TNF-alpha potentiates the effect of interleukin-1beta (IL-1beta) or TxA2 receptor activation on pulmonary microvascular permeability. Lungs harvested from Wistar rats were perfused ex vivo with Krebs-Henseleit buffer containing 0, 10, 100, or 1000 ng/mL recombinant rat TNF-alpha. Twenty minutes later pulmonary microvascular permeability was determined by measuring the capillary filtration coefficient (Kf) using a gravimetric technique. The effect of TNF-alpha (100 ng/mL) on pulmonary Kf was compared with that of lungs exposed to LPS (400 microg/mL; E. coli 0111:B4) or a TxA2 receptor agonist (U-46619; 7 x 10(-8)). In other experiments, perfused lungs were exposed to TNF-alpha plus IL-1beta (1 ng/mL) or TNF-alpha plus U-46619 after which Kf was measured. Exposure of ex vivo buffer perfused lungs to 10-1000 ng/mL TNF-alpha had no effect on Kf whereas LPS and U-46619 was associated with a two- and six-fold increase in Kf, respectively (P < 0.05). The Kf of lungs exposed to TNF-alpha plus IL-1 was similar to that of lungs exposed to TNF-alpha alone. Lastly, the Kf of lungs exposed to TNF-alpha plus U-46619 was not different than that of lungs exposed to U-46619 alone. In conclusion, TNF-alpha at least when administered for a relatively brief period of time does not affect microvascular permeability in an isolated, buffer-perfused lung model. PMID- 12095139 TI - Effects of dopamine on systemic and regional blood flow and metabolism in septic and cardiac surgery patients. AB - Dopamine is used in the clinical setting to support cardiac output and blood pressure and to improve diuresis. Experimental studies suggest that dopamine may reduce splanchnic perfusion and redistribute blood flow locally. To assess the effects of dopamine on splanchnic perfusion, we used dopamine to increase cardiac output by 25% in nine septic patients and 11 patients after cardiac surgery. Systemic (pulmonary artery catheter) and splanchnic (hepatic vein catheter and dye dilution) hemodynamics and oxygen transport were measured at baseline and 90 min after increasing the cardiac output. Dopamine infusion [in cardiac surgery patients 4.2 (1.4-8.5) microg x kg(-1) x min(-1) (median, range) and in septic patients 4.0 (2.1-9.0) microg x kg(-1) x min(-1)] increased splanchnic blood flow in cardiac surgery patients from 0.61 (0.13) L x min(-1) x m(-2) to 0.82 (0.13) L x min(-1) x m(-2) [mean (standard deviation; SD); P = 0.018] and in septic patients from 0.91 (0.32) L x min(-1) x m(-2) to 1.12 (0.40) L x min(-1) x m(-2) (P = 0.038). Splanchnic oxygen consumption increased in cardiac surgery patients from 39 (5) mL x min(-1) x m(-2) to 46 (6) mL x min(-1) x m(-2) (P = 0.003) but decreased in septic patients from 61 (19) mL x min(-1) x m(-2) to 51 (17) L x min(-1) x m(-2) (p = 0.021). Because of the unexpected results, we compared these data post hoc with data obtained from another group of 15 septic patients with acute lung injury, where dobutamine was used to increase cardiac output in a similar design. Dobutamine in these patients [6.4 (4.2-9.5) microg x kg(-1) x min(-1)] increased splanchnic blood flow from 1.20 (0.44) L x min(-1) x m(-2) to 1.43 (0.57) L x min(-1) x m(-2) (P = 0.008), while splanchnic oxygen consumption did not change 72 (25) mL x min(-1) x m(-2) vs. 76 (22) mL x min(-1) x m(-2) (not significant)]. The reduction of splanchnic oxygen consumption by dopamine in sepsis suggests an impairment of hepatosplanchnic metabolism despite an increase in regional perfusion. The safety and indications of dopamine use in sepsis should be re-evaluated. PMID- 12095140 TI - The role of p38 map kinase in tumor necrosis factor-induced redistribution of vascular endothelial cadherin and increased endothelial permeability. AB - The mechanism by which tumor necrosis factor alpha (TNFalpha) increases endothelial permeability is unclear. Vascular endothelial (VE) cadherin (cadherin 5) is an important contributor to endothelial monolayer integrity. The purpose of our study was to determine the effect of TNFalpha on VE-cadherin cell-surface expression and to identify the signaling pathways involved in TNF-induced changes in cadherin expression. Human umbilical vein endothelial cell monolayer permeability was measured by enzyme-linked immunosorbent assay for biotin-labeled albumin. Immunofluorescence, laser confocal microscopy, and Western immunobloting were used to assess VE cadherin distribution. Mitogen-activated protein kinase (MAPK) activity was determined using functional kinase assays and was inhibited with the compounds SB202190 and PD98059. TNFalpha significantly increased permeability and induced p38 and ERK MAPK activation compared with controls (P < 0.05). These changes were associated with a loss of membrane-associated VE cadherin. Inhibition of p38 but not ERK MAPK significantly reduced the effect of TNFalpha on endothelial permeability and cell-surface VE cadherin expression. p38 MAP kinase activation appears to be an important upstream signaling event associated with increased endothelial permeability and vascular endothelial cadherin redistribution. PMID- 12095141 TI - The effect of intestinal ischemia and reperfusion injury on ICAM-1 expression, endothelial barrier function, neutrophil tissue influx, and protease inhibitor levels in rats. AB - Multiple organ dysfunction syndrome (MODS) is mediated by complex mechanisms in which interactions between activated leukocytes and endothelial cells play a central role. ICAM-1 (intercellular adhesion molecule-1) mediates firm adhesion and transendothelial migration of activated leukocytes from postcapillary venules into the tissue. The present study evaluated the ICAM-1 expression in various organs after 40 min of intestinal ischemia and 1, 3, 6, 12 h of reperfusion (I/R) in the rat, using a dual monoclonal antibody technique (n = 36). Endothelial barrier permeability, using the vascular leakage of radiolabeled human serum albumin was also assessed (n = 12). Neutrophil sequestration in the lungs was quantitated by myeloperoxidase activity and plasma protease inhibitor levels were measured with electroimmunoassay. Significant regional differences were found in ICAM-1 expression between organs, both constitutively and after I/R-injury. The highest constitutive levels were observed in the liver and lungs, followed by the kidneys. The constitutive ICAM-1 expression in the intestines and in the heart was about 1/20 compared with that found in the liver and lungs. The brain and muscle had levels of about 1/150 of that in the liver and lungs. After intestinal I/R, significant increases (17-45%) were found in the lungs, intestines, brain, heart, and muscle. Albumin leakage index (ALI) in all examined organs and myeloperoxidase activity in the lungs increased after I/R-injury. Serum levels of albumin and most protease inhibitors decreased significantly after I/R challenge. Intestinal I/R results in an increase of systemic ICAM-1 expression with marked organ variability. The upregulation of ICAM-1 could represent a crucial step in the adherence- and migration process of activated leukocytes and potentially in the development of tissue injury. PMID- 12095142 TI - Peroxynitrite enhances the ability of Salmonella dublin to invade T84 monolayers. AB - In the intestine, epithelial cells continually produce and secrete low levels of nitric oxide (NO). Salmonella sp. invade epithelium by responding to environmental stimuli. The aims of this study were to determine the effect of reactive nitrogen intermediates (RNIs) on S. dublin and S. typhimurium growth and invasion of T84 epithelial monolayers. Intracellular NO formation was inhibited by 7-nitroindazole (7-NI) or N(G)-monomethyl-L-arginine, monoacetate (L-NMMA); extracellular NO and peroxynitrite were scavenged with ferro-hemoglobin or urate. The effect of authentic peroxynitrite (ONOO-); 3-morpholino-sydnonimine (SIN-1), which releases ONOO- via NO and superoxide; spermine NONOate, which releases only NO; or superoxide generated by xanthine oxidase and pterin on S. dublin and S. typhimurium growth and invasion were examined. Inhibition of NO synthesis and scavenging of extracellular NO or peroxynitrite reduced S. dublin invasion into T84 monolayers and enhanced bacterial growth. Pre-exposure of S. dublin to ONOO- and SIN-1 increased subsequent bacterial invasion into T84 monolayers. Conversely, exposure of bacteria to spermine NONOate or superoxide did not affect S. dublin invasion. In contrast, S. typhimurium invasion was not affected by pre treatment with NO donors. In conclusion, exposure of S. dublin to ONOO- enhances the ability of the bacteria to invade epithelial cells. These results suggest that luminal ONOO- may have a novel role as an extracellular signal between invasive bacteria and epithelial cells. PMID- 12095144 TI - Cell cycle entry of hematopoietic stem and progenitor cells controlled by distinct cyclin-dependent kinase inhibitors. AB - The therapeutic promise of hematopoietic stem cells in medicine has been expanded as broader differentiation potential of the cells has gained experimental support. However, hurdles for stem cell manipulation in vitro and tissue regeneration in vivo remain because of lack of the molecular biology of the stem cells. In particular, elucidating the molecular control of cell cycle entry is necessary for rational stem cell expansion strategies. Understanding how the stem and progenitor cell populations are controlled by negative regulators of cell cycle entry may provide one basis for manipulating these cells. In this mini review, we focus on the rationale of targeting the cyclin-dependent kinase inhibitors (CKIs) in stem cell biology. Two CKI members, p21(Cip1/Waf1) (p21) and p27kip1 (p27), have been shown to govern the pool sizes of hematopoietic stem and progenitor cells, respectively. Of note, their inhibitory roles in primitive hematopoietic cells are distinct from the action of the inhibitory cytokine, transforming growth factor-beta1 (TGF-beta1). Therefore, the distinct roles of p21, p27, and TGF-beta1 in hematopoietic cells offer attractive targets for specific manipulation of the stem or progenitor cell populations in therapeutic strategies. PMID- 12095143 TI - The notch pathway: modulation of cell fate decisions in hematopoiesis. AB - The hematopoietic system is maintained by a rare population of hematopoietic stem cells (HSC) that are thought to undergo self-renewal as well as continuously produce progeny that differentiate into the various hematopoietic lineages. However, the mechanisms regulating cell fate choices by HSC and their progeny have not been understood. Results of most studies support a stochastic model of cell fate determination in which growth factors support only the survival or proliferation of the progeny specified along a particular lineage. In other developmental systems, however, Notch signaling has been shown to play a central role in regulating fate decisions of numerous types of precursors, often inhibiting a particular (default) pathway while permitting self-renewal or differentiation along an alternative pathway. There is also accumulating evidence that the Notch pathway affects survival, proliferation, and cell fate choices at various stages of hematopoietic cell development, including the decisions of HSC to self-renew or differentiate and of common lymphoid precursors to undergo T- or B-cell differentiation. These data suggest that the Notch pathway plays a fundamental role in the development and maintenance of the hematopoietic system. PMID- 12095147 TI - Abnormal NK cell lymphocytosis detected after splenectomy: association with repeated infections, relapsing neutropenia, and persistent polyclonal B-cell proliferation. AB - We report the case of a boy with hereditary spherocytosis who presented with mild microcytic hypochromic anemia and recurrent leg ulcers that had been present since childhood. Chronic natural killer (NK) cell and B-cell lymphocytosis was detected 1 year after therapeutic splenectomy during investigation of recurrent episodes of neutropenia and persistent lymphocytosis. NK cells proved to be abnormal at immunophenotyping studies, and B-cells were polyclonal and displayed a normal immunophenotype. Genotypic analysis of T-cell receptor (TCR)-beta and TCR-gamma genes showed a germ-line pattern. The clinical course of this patient was characterized by multiple pulmonary infections and amygdalitis. We discuss the potential roles of persistent immune stimulation due to chronic hemolysis and severe leg ulcers and of splenectomy in the origin of NK cell lymphocytosis and the relationship between NK cells and recurrent infections, relapsing neutropenia, and polyclonal B-cell response. PMID- 12095145 TI - Transcriptional regulation of myelopoiesis. AB - A common myeloid progenitor gives rise to both granulocytes and monocytes. The early stages of granulopoiesis are mediated by the C/EBPalpha, PU.1, RAR, CBF, and c-Myb transcription factors, and the later stages require C/EBPepsilon, PU.1, and CDP. Monocyte development requires PU.1 and interferon consensus sequence binding protein and can be induced by Maf-B, c-Jun, or Egr-1. Cytokine receptor signals modulate transcription factor activities but do not determine cell fates. Several mechanisms orchestrate the myeloid developmental program, including cooperative gene regulation, protein:protein interactions, regulation of factor levels, and induction of cell cycle arrest. PMID- 12095146 TI - Molecular control of megakaryopoiesis and thrombopoiesis. AB - Megakaryopoiesis and subsequent thrombopoiesis occur through complex biologic steps: megakaryocyte precursors that developed from hematopoietic stem cells initially proliferate, then differentiate into mature polyploid megakaryocytes, and finally release platelets. Although a number of growth factors can augment megakaryopoiesis in vitro, thrombopoietin is a physiologic and the most potent regulator of megakaryopoiesis in vitro and in vivo. Thrombopoietin induces the growth of megakaryocyte precursors through activation of multiple signaling cascades, including Ras/mitogen-activated protein kinase (MAPK), signal transducers and activators of transcription 5 (STAT5), phosphatidylinositol 3 kinase (PI3-K)/Akt, and protein kinase C, whereas it induces megakaryocytic maturation primarily through the Ras/MAPK pathway. During the maturation step, megakaryocytes undergo polyploidization characterized by repeated rounds of DNA replication without concomitant cell division. During these rounds of replication, cytokinesis is neglected because of the down-regulated expression of AIM-1, and DNA replication occurs through the increased expression of D-type cyclins. As for transcriptional regulation during megakaryopoiesis, GATA-1 plays a central role in the lineage commitment of hematopoietic stem cells toward erythroid/megakaryocytic lineage and subsequent maturation. p45 NF-E2 is essential for platelet release from terminally differentiated megakaryocytes. At present, mutations of GATA-1, AML1, and HOXA11 genes have been found in hereditary diseases accompanying thrombocytopenia among humans. PMID- 12095148 TI - Effect of continuous subcutaneous administration of a low dose of G-CSF on stem cell mobilization in healthy donors: a feasibility study. AB - We investigated the effects of low-dose granulocyte colony-stimulating factor (G CSF) on the mobilization of stem cells in 6 healthy subjects. When G-CSF was administered by continuous subcutaneous infusion at a rate of 72 microg/day for 5 days, the numbers of white blood cells and granulocytes rapidly increased to maximal levels. CD34+ cells were mobilized to the peripheral blood in 3 days, and the maximal level was reached 4 or 5 days after the start of treatment. We attempted to determine whether the levels of mobilized stem cells that we could obtain using this method would be sufficient for peripheral blood stem cell transplantation. Two of the 6 subjects complained of mild bone pain 4 or 5 days after the start of treatment, but the pain did not affect their daily activities. Only 1 abnormal result (for serum alkaline phosphatase) was found in the laboratory data. The present preliminary results have provided us with a framework for a prospective study comparing low-dose continuous infusion with conventional mobilization procedures. PMID- 12095149 TI - Long-term follow-up of allogeneic bone marrow transplantation after reduced intensity conditioning in patients with chronic myelogenous leukemia in the chronic phase. AB - Although allogeneic transplantation is a curative therapy for chronic myelogenous leukemia (CML), treatment-related mortality is still a major cause of death after transplantation, especially in older patients. We investigated the safety and efficacy of reduced-intensity conditioning consisting of low-dose (600 cGy) total body irradiation and cytosine arabinoside (1 g/m2) together with a continuous infusion of granulocyte colony-stimulating factor and cyclophosphamide (120 mg/kg) in patients with CML in the chronic phase. Fractionated splenic irradiation (5 Gy) was also administered as part of the conditioning treatment. Eight patients older than 40 years underwent allogeneic bone marrow transplantation from an HLA-matched sibling following this conditioning. Regimen related toxicities (equal to or greater than grade III) were not observed. Rapid restoration of 100% donor chimerism was confirmed by fluorescence in situ hybridization methods in 5 sex-mismatched transplant recipients. One patient died from severe acute graft-versus-host disease and another from Pneumocystis carinii pneumonia early in the course of transplantation. A sustained engraftment was achieved in 5 long-term survivors; in 1 case, the graft was rejected but the Philadelphia chromosome and BCR/ABL-negative autologous hemopoiesis were restored. After a minimum follow-up period of 60 months, 6 patients, including the patient with restored autologous hemopoiesis, were still alive and in remission with 100% donor chimerism. Six years after the transplantation, 1 patient experienced a cytogenetic relapse, which was successfully treated with donor lymphocyte infusions. In summary, this reduced-intensity conditioning resulted in a cure with markedly reduced regimen-related toxicities in this relatively older cohort of patients with CML. PMID- 12095150 TI - An addition to geographic hematology: chronic myeloproliferative diseases are infrequent in Mexican Mestizos. AB - BACKGROUND: The chronic myeloproliferative diseases (CMPDs) include chronic myelogenous leukemia (CML), primary (essential) thrombocythemia (PT), agnogenic myeloid metaplasia (AMM), and polycythemia vera (PV). Certain hematological malignancies have a different prevalence in our country than in countries with Caucasian populations. Data indicate that the prevalence of CML in our country is similar to that found in Caucasians; however, the prevalence of the other CMPDs has not been studied. METHODS: In a total of 8069 individuals studied between June 1983 and March 2001 in the Centro de Hematologia y Medicina Interna de Puebla, we assessed the prevalence of CML, PT, AMM, and PV. Some of the clinical features of these individuals were also assessed. RESULTS AND CONCLUSIONS: Forty nine patients with CML, 14 with PT, 7 with AMM, and 3 with PV were identified. The clinical presentations of these CMPDs were not different from those described in Caucasians. CONCLUSIONS: We found that CML was more than 3 times more frequent than PT, that both PV and AMM were exceptional, and that PT, AMM, and PV were significantly less frequent in Mexican than in Caucasian populations (P < .01). PMID- 12095151 TI - Fusion of MLL and MSF in adult de novo acute myelomonocytic leukemia (M4) with t(11;17)(q23;q25). AB - The MLL gene at chromosome band 11q23 is frequently rearranged and fused to partner genes in acute leukemias. Previously, the MSF gene, also called AF17q25, has been cloned as a fusion partner of the MLL gene in therapy-related or infant acute myelogenous leukemias with t(11;17)(q23;q25). MSF belongs to the septin family of proteins, which includes other MLL fusion partners, hCDCrel1 and Septin 6, and has also been implicated in the pathogenesis of human ovarian tumor and murine T-cell lymphoma. We describe here a 64-year-old man with de novo acute myelomonocytic leukemia (French-American-British subtype M4) with t(11;17)(q23;q25). His leukemia was successfully induced into a first remission, which, however, lasted only briefly. A second remission was never attained, and the patient died of sepsis 16 months after the diagnosis of leukemia. Examination of his leukemic cells at diagnosis revealed an MLL gene rearrangement, by Southern blotting, and an MLL-MSF fusion transcript, by the reverse transcriptase polymerase chain reaction (RT-PCR) method. Sequence analysis of the RT-PCR product further revealed that MLL exon 5 was fused in-frame to MSF exon 3. Further clinical and molecular analyses of acute leukemias with the MLL-MSF transcript may shed more light on the clinical characteristics and molecular mechanisms of the MLL-septin type leukemias. PMID- 12095152 TI - Activation of extracellular signal-regulated kinase through B-cell antigen receptor in B-cell chronic lymphocytic leukemia. AB - B-cell chronic lymphocytic leukemia (B-CLL) cells express on their surface membranes immunoglobulin (Ig) M or IgD, both of which normally function as B-cell antigen receptors (BCRs). However, in contrast to normal B-cells, in B-CLL cells several important signaling pathways, such as the activation of protein tyrosine kinase via BCR, are defective. We have examined whether the activities of mitogen activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), p38 MAPK, and Akt kinase, are functional in B-CLL cells, because these kinases play critical roles in activation in response to BCR stimulation, tumor cell growth, and survival. In B CLL cells, BCR cross-linking neither induced activation nor enhanced the activities of Lyn, Syk, p21ras, JNK, p38 MAPK, or Akt kinases, whereas p38 MAPK and Akt were constitutively active. In contrast, BCR cross-linking resulted in ERK activation, although the activation in quiescent cells was case dependent. These results suggest that some signaling pathways, such as the activation of ERK through BCR, are functional in B-CLL cells despite the extensive impairment of signaling pathways. PMID- 12095153 TI - Pure red cell aplasia developing into myeloproliferation with myelodysplasia and subsequent leukemia after cyclosporin A therapy. AB - We describe a very rare case of a patient who presented with red cell aplasia that later developed into myeloproliferation with myelodysplasia and eventually leukemia. A 63-year-old man presented with anemia and reticulocytopenia in May 1997. A bone marrow examination revealed erythroid aplasia with normal production of myeloid cells and megakaryocytes with a normal karyotype. After the diagnosis of pure red cell aplasia was made, the patient was treated with prednisolone and then with cyclosporin A (CyA). Two weeks after the initiation of CyA treatment, the peripheral reticulocyte count began to increase with a regrowth of erythroid cells in the bone marrow. Meanwhile, the peripheral white blood cell and platelet counts also increased to more than 10,000/microL and 1,000,000/microL, respectively. Examination of a bone marrow aspirate in December 1997 revealed myelodysplastic changes with trisomy 8. Despite the discontinuation of CyA and the administration of 1-beta-D-arabinofuranosylcytosine stearyl monophosphate, leukemia developed in August 1998. In September 1998, the patient died of sepsis during a neutropenic period that followed remission-induction therapy. In the mechanism of pathogenesis, CyA may induce upon pure red cell aplasia a secondary myeloproliferative disorder with myelodysplasia and leukemia. An alternative possibility is that CyA reduces autoimmune-mediated suppression of the underlying stem cell disorder and that the result of this reduction is the manifestation of myeloproliferation and leukemia. PMID- 12095154 TI - Flexible low-intensity combination chemotherapy for elderly patients with acute myeloid leukemia. AB - Twenty-five patients aged 57 to 88 years (median, 70 years) with acute myeloid leukemia were treated with a flexible low-intensity treatment regimen comprising mitozantrone (mitoxantrone) 6 mg/m2 administered by intravenous infusion x3 days, cytarabine 10 mg/m2 subcutaneously every 12 hours x7 to 14 days, and etoposide 100 mg orally x7 to 14 days. Seventeen of these patients had a preexisting myelodysplastic syndrome. The clinical response was correlated to the results of cytogenetic studies (23 patients) and of viability studies of leukemic blasts (7 patients). Eleven of the 25 patients achieved complete remission (CR), 8 achieved partial remission (PR), and 4 showed no response. There was 1 toxic death, and 1 patient died soon (1 week) after presentation. Treatment was well tolerated. Although myelotoxicity occurred regularly, the recovery time was < or = 3 weeks for most of the responding patients. Duration of survival for patients who had CR has ranged from 4+ to 43+ months and for patients who had PR, 3 to 16 months. Irrespective of the remission status (CR or PR), responding patients with favorable (n = 1) or intermediate (n = 10) cytogenetic findings had a significantly better survival time (median, 14 months) than did those with unfavorable (n = 7) cytogenetic findings (median, 5 months). In vitro studies showed a progressive reduction in the number of circulating blasts. The number of viable blasts 3 days after initiation of therapy appeared to give an early indication of clinical response. Treatment with a flexible low-intensity protocol seems to achieve results comparable with those reported for intensive antileukemia therapy and has much less toxicity. PMID- 12095155 TI - Resveratrol, a natural product derived from grapes, is a new inducer of differentiation in human myeloid leukemias. AB - A natural product, resveratrol (3,4,40-trihydroxy-trans-stilbene), a phytoalexin found in grapes and other food products, is known as a cancer chemopreventive agent. We studied the in vitro biological activity of this compound by examining its effect on proliferation and differentiation in myeloid leukemia cell lines (HL-60, NB4, U937,THP-1, ML-1, Kasumi-1) and fresh samples from 17 patients with acute myeloid leukemia. Resveratrol (20 microM, 4 days) alone inhibited the growth in liquid culture of each of the 6 cell lines. Resveratrol (10 microM) enhanced the expression of adhesion molecules (CD11a, CD11b, CD18, CD54) in each of the cell lines except for Kasumi-1. Moreover, resveratrol (25 microM, 4 days) induced 37% of U937 cells to produce superoxide as measured by the ability to reduce nitroblue tetrazolium (NBT). The combination of resveratrol (10 microM) and all-trans-retinoic acid (ATRA) (50 nM, 4 days) induced 95% of the NB4 cells to become NBT-positive, whereas <1% and 12% of the cells became positive for NBT after a similar exposure to either resveratrol or ATRA alone, respectively. In U937 cells exposed to resveratrol (25 microM, 3 days), the binding activity of nuclear factor-kappaB (NFkappaB) protein was suppressed. Eight of 19 samples of fresh acute leukemia cells reduced NBT after exposure to resveratrol (20 microM, 4 days). Taken together, these findings show that resveratrol inhibits proliferation and induces differentiation of myeloid leukemia cells. PMID- 12095156 TI - Synergistic growth inhibition of YM529 with biologic response modifiers (BRMs) in myeloma cells. AB - Bisphosphonates (BPs) are effective in the management of bone disease in patients with multiple myeloma. Recent reports have suggested that they may also have an antitumor activity. YM529 is a new synthetic BP with more than 1000 times the bone resorption inhibitory activity of pamidronate. To clarify the direct effects of YM529 on myeloma cells, the cell proliferation and cell cycle perturbation were analyzed using 12 myeloma cell lines established in our laboratory. The growth inhibition was dose dependent. The cells accumulated in [2n<<4n] of the cell cycle and subsequently formed an apoptotic sub-G1 fraction. Combined treatment with all-trans retinoic acid, thalidomide, or interferon-alpha enhanced the growth inhibitory effects of YM529 on these cells. However, there were no remarkable effects of YM529 on the messenger RNA expression for angiogenic factors, cell cycle regulators, or cytokines related to myeloma cells. These results indicate that YM529 is beneficial not only to bone lesions but also for its direct antitumor effects on myeloma cells. PMID- 12095157 TI - A comparative double-blind randomized trial of activated protein C and unfractionated heparin in the treatment of disseminated intravascular coagulation. AB - A randomized prospective double-blind trial was performed to compare the safety and efficacy of human activated protein C (APC) and unfractionated heparin for the treatment of disseminated intravascular coagulation (DIC). One hundred thirty two patients with DIC were enrolled in this study: 63 patients received APC (12.5 U [2.5 microg]/kg body wt per hour) and 69 patients received heparin (8 U/kg body wt per hour) by intravenous infusion for 6 days. Forty-nine APC-treated patients and 55 heparin-treated patients were evaluated for efficacy, and 52 APC-treated patients and 55 heparin-treated patients were evaluated for safety. The 2 groups were similar with respect to sex, age, body weight, underlying diseases, and coagulation/fibrinolysis parameters before treatment. Aggravation of bleeding was seen after treatment in 8 patients receiving heparin, but in none of the patients receiving APC. The number of patients who showed alleviation of bleeding was significantly higher in the APC group than the heparin group (P = .009). The effects on DIC-related organ dysfunction were not significantly different between the 2 groups. Fibrinogen-fibrin degradation products, D-dimer, thrombin antithrombin complex (TAT), and plasmin-plasmin inhibitor complex (PIC) were all significantly decreased by treatment in both groups. Fibrinogen, protein C, and antithrombin were significantly increased in the APC group, whereas only protein C was significantly increased in the heparin group. Platelet count in the nonleukemic group was significantly increased in those patients receiving APC but not increased in those patients receiving heparin. Improvement of coagulation/fibrinolysis was assessed by scoring 4 parameters (soluble fibrin monomers, D-dimer, TAT, and PIC), and the results indicated that the APC group showed significantly greater improvement than the heparin group (P = .046). There was, however, no significant difference in the rate of complete recovery from DIC between the 2 groups. The rate of death from any cause within 28 days after treatment was 20.4% in the APC group, significantly lower than the 40% death rate observed in the heparin group (P < .05). There were no severe adverse events in either group. These results suggest that APC in a relatively small dosage can improve DIC more efficiently than can heparin, without increasing bleeding, and may be a better alternative. PMID- 12095158 TI - PLAIDD, a type II death domain protein that interacts with p75 neurotrophin receptor. AB - We describe the cloning and characterization of a rat single transmembrane protein that is homologous to the common neurotrophin receptor p75NTR in its death domain and the transmembrane region but dissimilar outside these regions. We have dubbed this protein PLAIDD, for p75-like apoptosis-inducing death domain protein. PLAIDD messenger RNA, which is ubiquitously distributed, is highly expressed in the embryo, but downregulated in adult tissues. Alternative splicing within the extracellular region of PLAIDD generates four RNA species, but only two of them are translated, PLAIDD_L and PLAIDD_S (long and short isoforms, respectively). While the amino acid sequence of the intracellular region of PLAIDD displays 41% identity with the intracellular region of p75NTR, the extracellular region of PLAIDD does not reveal any homology with p75NTR. Overexpression of each isoform of PLAIDD led to cytotoxicity in superior cervical ganglion neurons and in human embryonic kidney 293T cells. Both isoforms of PLAIDD could be co-immunoprecipitated with p75NTR, suggesting an interaction between these molecules. PMID- 12095159 TI - Zinc inhibits the nuclear translocation of the tumor suppressor protein p53 and protects cultured human neurons from copper-induced neurotoxicity. AB - High concentrations of the trace metal zinc (Zn) have previously been shown to provide transient protection of cells from apoptotic death. The molecular mechanisms responsible for this protection are not known. Thus, this work explored the ability of Zn to protect human neurons in culture (NT2-N) from Cu mediated death and tested the hypotheses that the tumor-suppressor protein p53 plays a role in Cu-induced neuronal death and is part of the mechanism of Zn protection. Copper toxicity (100 microM) resulted in significant apoptotic neuronal death by 12 h. Addition of 100 microM Zn to Cu-treated cells increased neuronal death. However, the addition of 700 microM Zn to Cu-treated cells resulted in neuronal viability that was not different from untreated controls through 24 h. p53 mRNA abundance, while increased by the addition of Cu and 100 microM Zn, was decreased to 50% of control with the addition of 500 microM Zn in Cu-treated cells, and to 10% of control with 700 microM Zn. Consistent with its role as a transcription factor, both Western analysis and immunocytochemistry showed significant increases in nuclear p53 protein levels in Cu toxicity. The role of p53 in Cu-mediated apoptosis was further confirmed by elimination of apoptosis in Cu-treated cells that had been transfected with a dominant-negative p53 construct to prevent p53 expression. Furthermore, the addition of 500-700 microM Zn prevented the movement of p53 into the nucleus suggesting that Zn not only protects neurons from Cu toxicity by regulating p53 mRNA abundance but also by preventing the translocation of p53 to the nucleus. PMID- 12095160 TI - Cytochrome C and caspase-9 expression in Huntington's disease. AB - There is increasing evidence implicating apoptosis-mediated cell death in the pathogenesis of neurodegenerative diseases. One important event in the apoptotic cascade is the release of cytochrome c by mitochondria into the cytoplasm, activating caspase-9, leading to the subsequent activation of downstream executioner caspases. In the present study, we examined the distribution of cytochrome c and caspase-9 in Huntington's disease (HD) patients and in a transgenic model of HD (R6/2 line). Neuronal cytochrome c immunoreactivity increased with neuropathological severity in HD patients. Concomitant with this finding, Western-blot analysis showed a shift in the distribution of cytochrome c from the mitochondrial to the cytosolic fraction with incremental cytosolic expression associated with greater striatal degeneration. Active caspase-9 immunoreactivity was present in both HD striatal neurons and in Western blots of severe-grade specimens. Similar findings were observed in the R6/2 mice. There was a temporal increase in expression and shift of cytochrome c from the mitochondrial to the cytosolic fraction from 4-13 wk of age. Activated caspase-9 and caspase 3 activities were present only at endstage disease. Although the present results provide evidence that key components of the intrinsic mitochondrial apoptotic pathway are activated in both HD patients and a transgene murine model of HD, these phenomena are prominent in only severe neuropathological grades in HD patients and HD mice, suggesting that apoptosis may play a greater role in neuronal death at endstage disease. PMID- 12095163 TI - Effects of the ethanol extract of Cichorium intybus on the immunotoxicity by ethanol in mice. AB - Effects of the ethanol extract of Cichorium intybus (CIEE) on the immunotoxicity of ethanol (EtOH) were investigated in ICR mice. Mice were divided into four groups, and CIEE at dose of 300 mg/kg was orally administered to mice daily for 28 consecutive days, and normal mice were given vehicle. Mice treated with EtOH were given freely with 20% w/v EtOH solution. The results of this study are summarized as follows: The combination of CIEE and EtOH showed significant increases in the circulating leukocytes and the relative weights of liver, spleen and thymus, as compared with those in mice treated with EtOH alone. However, the body weight gain was not affected. Splenic plaque forming cells (PFC) and hemagglutination (HA) titers to sheep red blood cells (SRBC), and the secondary IgG antibody response to bovine serum albumin (BSA) were markedly enhanced by CIEE plus EtOH treatment as compared with the treatment of EtOH alone. In mice receiving the combination of CIEE and EtOH when compared with EtOH alone-treated mice, there were also significant increases in delayed-type hypersensitivity (DTH) reaction, phagocytic activity, natural killer (NK) cell activity and cell proliferation as well as interferony (IFN-gamma) secretion. In the case of interleukin-4 (IL-4) content, however, an insignificant induction observed by CIEE plus EtOH treatment. These findings indicate that the immunotoxicity induced by EtOH is significantly restored or prevented by CIEE treatment. PMID- 12095162 TI - Behavioral and neurological correlates of ALS-parkinsonism dementia complex in adult mice fed washed cycad flour. AB - Consumption of cycad seed products (Cycas circinalis) is one of the strongest epidemiological links to the Guamian neurological disorder amyotrophic lateral sclerosis-parkinsonism-dementia complex (ALS-PDC), however, the putative toxin which causes neurodegeneration has never been identified definitively. To reexamine this issue, 6-7-mo-old, male CD-1 mice were assessed for motor and cognitive behaviours during and following feeding with pellets made from washed cycad flour. Cycad-fed animals showed early evidence of progressive motor and cognitive dysfunctions. Neurodegeneration measured using TUNEL and caspase-3 labeling was found in neocortex, various hippocampal fields, substantia nigra, olfactory bulb, and spinal cord. In vitro studies using rat neocortex have identified toxic compounds in washed cycad flour that induce depolarizing field potentials and lead to release of lactate dehydrogenase (LDH), both blocked by AP5. High-performance liquid chromatography (HPLC)/mass spectrometry of cycad flour samples failed to show appreciable amounts of other known cycad toxins, cycasin, MAM, or BMAA; only trace amounts of BOAA were present. Isolation procedures employing these techniques identified the most toxic component as beta sitosterol beta-D-glucoside (BSSG). The present data suggest that a neurotoxin, or a toxic metabolite, not previously identified in cycad, is able to gain access to central nervous system (CNS) resulting in neurodegeneration of specific neural populations and in motor and cognitive dysfunctions. These data are consistent with a number of major features of ALS-PDC in humans. PMID- 12095161 TI - 5-HT2A receptor activation leads to increased BDNF mRNA expression in C6 glioma cells. AB - It has recently been suggested that an increase in brain-derived neurotrophic factor (BDNF) expression may mediate some of the therapeutic effect of antidepressant drugs, via their effects on the neurotransmitter 5 hydroxytryptamine (5-HT). However, because it is unclear whether 5-HT manipulations directly affect BDNF expression, we examined BDNF mRNA levels in C6 glioma cells following incubation with 5-HT using reverse transcription polymerase chain reaction (RT-PCR) and Northern blot analysis. Incubation of C6 glioma cells with 5-HT increased the BDNF mRNA expression approx twofold. The effect of 5-HT (100 microM) was inhibited by a 5-HT2A receptor antagonist (ketanserin; 1 microM). The RNA synthesis inhibitor (actinomycin D; 10 microg/mL), but not a protein synthesis inhibitor (cycloheximide; 0.5 microg/mL) blocked the effect of 5-HT. Furthermore, incubation of C6 glioma cells with EGTA (1 mM), a protein kinase inhibitor (staurosporine; 1 microM), the Ca2+ ATPase inhibitor thapsigargin (1 microM), or a calcium/calmodulin-dependent kinase inhibitor (KN 62; 1 microM) inhibited the response to 5-HT. Our data show that 5 HT increases de novo BDNF mRNA synthesis following direct activation of the 5 HT2A receptor, via a calcium-dependent and protein kinase-dependent pathway. PMID- 12095164 TI - Histamine H1 receptors are involved in mouse nasal allergic responses: a demonstration with H1 receptor-deficient mice. AB - The role of histamine H1 receptors in nasal allergic symptoms (sneezing and nasal rubbing) were studied using histamine H1 receptor-deficient mice. Intranasal instillation of histamine solution resulted in significant increases in sneezing and nasal rubbing in wild-type mice, whereas no increases were observed in histamine H1 receptor-deficient mice. The histamine H1 receptor agonist 2 pyridylethylamine induced sneezing and nasal rubbing in a dose-dependent-manner in wild-type mice, but no such increase was found in histamine H1 receptor deficient mice. On the other hand, the histamine H2 receptor agonist dimaprit did not increase sneezing and nasal rubbing in wild-type mice. Histamine H1 receptor antagonists such as chlorpheniramine and epinastine significantly inhibited nasal allergic symptoms caused by histamine, but the histamine H2 receptor antagonists cimetidine and famotidine showed no effect. No additional effects were observed by combined use of chlorpheniramine and cimetidine or famotidine compared with cimetidine or famotidine alone. These results suggested that histamine H1 receptors play an important role in nasal allergy symptoms induced by histamine. PMID- 12095165 TI - Inhibition of antigen specific lymphocyte proliferation and cytokine stimulation by peptidomimetic opioid compound. AB - In sequel to our preliminary observations with peptidomimetic opioid compounds, we have further investigated immunomodulatory activity of one peptidomimetic compound (Tyr-NH-CH2-CH2-O-Phe-NH2) with peripheral blood mononuclear cells (PBMCs) of healthy volunteers/tuberculosis patients. This peptidomimetic compound was evaluated for its effect on purified protein derivative (PPD) stimulated lymphocyte proliferation in vitro, production of Th1 and Th2 cytokines by ELISA and ribonuclease protection assay. Our study shows the immunosuppressive potential of above synthetic peptidomimetic compound. This compound inhibited PPD stimulated human lymphocyte proliferation and this inhibition was reversed by opioid receptor antagonist, naloxone. Its immunosuppressive effect was further demonstrated by inhibition of interleukin-9 (IL-9), IL-10 but failed to influence IL-2, IL-15 and interferon-y (IFN-gamma) in PPD stimulated human PBMCs. PMID- 12095166 TI - Side-stream cigarette smoke accentuates immunomodulation during murine AIDS. AB - Side-stream cigarette smoke has become a hotly debated social, political, and scientific health and safety issue for nonsmokers. The harmful influences of side stream cigarette smoke on human health are its adverse effects on the immune system, especially when already compromised by other agents. Acquired immune deficiency syndrome (AIDS) is a clinical disorder caused by human immunodeficiency virus (HIV). To facilitate studies, murine AIDS was induced in C57BL/6 mice by LP-BM5 murine leukemia virus infection, which mimics human AIDS. After 2 weeks of retroviral infection, the mice were exposed to side-stream cigarette smoke for 30 min, 5 days/week for 12 weeks using a side-stream cigarette smoke exposure system. Murine retrovirus infection reduced the in vitro proliferation of T lymphocytes stimulated by concanavalin A, increased the release of pro-inflammatory cytokine interleukin-6 (IL-6) tumor necrosis factor alpha (TNF-alpha), increased the hepatic lipid peroxidation and decreased the alpha-tocopherol levels in liver, lung and heart. Concomitant side-stream cigarette smoke exposure for 12 weeks further inhibited the proliferation of T cells, increased the release of TNF-alpha, IL-6 cytokines and enhanced the hepatic lipid peroxidation from retrovirus infected mice. The loss of alpha tocopherol was also further enhanced by side-stream cigarette smoke exposure during retrovirus infection. Our conclusions are that side-stream cigarette smoke induced increasing oxidative stress, reducing nutrient concentrations and suppressing immune function could make mice with murine AIDS more susceptible to opportunistic infections, potentially accelerating murine AIDS progression. Thus, the reduction of side-stream cigarette smoke exposure is an important health issue in AIDS patients to improve the quality and quantity of their lives. PMID- 12095167 TI - Resveratrol modulates rat macrophage functions. AB - This study investigated the effects of trans-resveratrol (trans-3,4',5 trihydroxystilbene, RESV), a natural polyphenol from grapes with known antioxidant activity, on the respiratory-burst responses and phagocytic activity of rat macrophages. RESV at concentrations of 1-10 microM significantly and dose dependently inhibited (a) the extracellular production of reactive oxygen intermediates (ROls) by resident peritoneal macrophages stimulated with phorbol 12-myristate 13-acetate (PMA) (a potent activator of protein kinase C, PKC) and (b) intracellular production of ROIs after opsonin-independent phagocytosis of Kluyveromyces lactis cells. Over the 10-100 microM concentration ranges, RESV likewise inhibited the production of reactive nitrogen intermediates (RNIs) by macrophages stimulated with thioglycollate. RESV concentrations above 10 microM also dose-dependently inhibited the phagocytosis of K. lactis cells. The results obtained demonstrate that RESV is a potent inhibitor of the antipathogen responses of rat macrophages and, thus, suggest that this agent may have applications in the treatment of diseases involving macrophage hyperresponsiveness. PMID- 12095169 TI - Efficacy of vaccination with plasmid DNA encoding for HER2/neu or HER2/neu-eGFP fusion protein against prostate cancer in rats. AB - Despite early diagnosis and improved therapy, 31,500 men will die from prostate cancer (PC) this year. The HER2/neu oncoprotein is an important effector of cell growth found in the majority of high-grade prostatic tumors and is capable of rendering immunogenicity. The antigenicity of this oncoprotein might prove useful in the development of PC vaccines. Our goal is to prove the principle that a single DNA vaccine can provide reliable immunity against PC in the MatLyLu (MLL) translational tumor model. The parental rat MatLyLu PC cell line expresses low to moderate levels of the rat neu protein. To simulate in vivo human PC, MatLyLu cells were transfected with a truncated sequence of human HER2/neu cDNA cloned into the pCI-neo vector. This HER2/neu cDNA sequence encodes the first 433 amino acids of the extracellular domain (ECD). MatLyLu cells were also transfected with the same HER2/neu cDNA sequence cloned into the N1-terminal sequence of EGFP reporter gene to produce a fusion protein. The partial ECD sequence of HER2/neu includes five rat major histocompatibility (MHC)-II-restricted peptides with complete human-to-rat cross-species homology. The HER2/neu protein overexpression was documented by Western Blot analysis, and the expression of fusion protein was monitored by confocal microscopy and fluorimetry. Vaccination with a single injection of HER2/neu cDNA protected 50% of animals against HER2/neu-MatLyLu tumors (P < 0.01). When the tumor cells were engineered to express HER2/neu-EGFP fusion protein, the antitumor immunity was enhanced, as following vaccination with HER2/neu-EGFP cDNA, 80% of these rats rejected HER2/neu-EGFP-MatLyLu (P<0.001). Both vaccines induced HER2/neu-specific antibody titers. Rats vaccinated with EGFP-cDNA rejected 80% of EGFP-MatLyLu tumors and, interestingly, 40% of HER2/neu-MatLyLu tumors. None of the cDNA vaccines induced immunity against parental MatLyLu cells. Our data clearly demonstrate that a single injection of HER2/neu-EGFP cDNA is a very effective vaccine against PC tumors expressing the cognate tumor-associated antigen (TA). The antitumor immunity is significantly more pronounced if the tumors express xenogeneic HER2/neu-EGFP fusion protein as opposed to only the syngeneic HER2/neu oncoprotein. Our data suggests that the HER2/neu-EGFP-MatLyLu tumor is a potential animal tumor model for investigating therapeutic vaccine strategies against PC in vivo and demonstrates the limitations of a cDNA vaccine only encoding for MHC-II restricted HER2/neu-ECD sequence peptides. PMID- 12095168 TI - Increased efficiency of Lewis lung carcinoma chemotherapy with a macrophage stimulator--yeast carboxymethyl glucan. AB - The efficiency of chemotherapy of Lewis lung carcinoma with cyclophosphamide was affected by administration of the water-soluble yeast polysaccharide derivative- carboxymethylated (1 --> 3)-beta-D-glucan (CMG)-a well-known macrophage stimulator. It was found that while cyclophosphamide showed 57% growth inhibition of the intramuscular tumor implants in comparison with the control group, its combined administration with CMG led to 75-90% inhibition. Similarly, increased inhibition of occurrence of lung metastases (up to 92-94%) was observed using the combined application of the two compounds. The stimulatory effect of CMG is not associated with the changed cellularity of peripheral blood, but is rather due to the obviously increased concentration of the intracellular inhibitor of cysteine proteases-stefin A and cystatin C in tumor tissue. PMID- 12095170 TI - Modulation of rat macrophage function by the Mangifera indica L. extracts Vimang and mangiferin. AB - Vimang is an aqueous extract of Mangiferia indica L., traditionally used in Cuba as an anti-inflammatory, analgesic and antioxidant. In the present study, we investigated the effects of Vimang and of mangiferin (a C-glucosylxanthone present in the extract) on rat macrophage functions including phagocytic activity and the respiratory burst. Both Vimang and mangiferin showed inhibitory effects on macrophage activity: (a) intraperitoneal doses of only 50-250 mg/kg markedly reduced the number of macrophages in peritoneal exudate following intraperitoneal injection of thioglycollate 5 days previously (though there was no significant effect on the proportion of macrophages in the peritoneal-exudate cell population); (b) in vitro concentrations of 0.1-100 microg/ml reduced the phagocytosis of yeasts cells by resident peritoneal and thioglycollate-elicited macrophages; (c) in vitro concentrations of 1-50 microg/ml reduced nitric oxide (NO) production by thioglycollate-elicited macrophages stimulated in vitro with lipopolysaccharide (LPS) and IFNgamma; and (d) in vitro concentrations of 1-50 microg/ml reduced the extracellular production of reactive oxygen species (ROS) by resident and thioglycollate-elicited macrophages stimulated in vitro with phorbol myristate acetate (PMA). These results suggest that components of Vimang, including the polyphenol mangiferin, have depressor effects on the phagocytic and ROS production activities of rat macrophages and, thus, that they may be of value in the treatment of diseases of immunopathological origin characterized by the hyperactivation of phagocytic cells such as certain autoimmune disorders. PMID- 12095171 TI - Inmunoferon, a glycoconjugate of natural origin, regulates the liver response to inflammation and inhibits TNF-alpha production by an HPA axis-dependent mechanism. AB - In this report, we have examined the mechanism of action of Inmunoferon, a patented glycoconjugate (GC) of natural origin with immunomodulatory properties, in the regulation of TNF-alpha expression induced by LPS challenge in a rodent model. GC was found to be dependent on a normal HPA response to exert its regulatory effect on TNF-alpha expression. Adrenalectomized mice were unresponsive to the drug in terms of TNF-alpha levels after LPS challenge, whereas control mice showed lower serum TNF-alpha levels when treated with GC than vehicle-treated animals. GC treatment also induced the expression of acute phase proteins but it did not alter the normal metabolism or viability of hepatic cells. These data suggested the employment of GC as a novel adjuvant during antibacterial treatment without disadvantages such as side effects in the metabolism of the liver, thus, acting as an enhancer of the host response against infection. PMID- 12095172 TI - Detrimental implication of B1 receptors in myocardial ischemia: evidence from pharmacological blockade and gene knockout mice. AB - OBJECTIVE: The aim of this study was to evaluate the contribution of kinin B1 receptors in myocardial ischemia using both pharmacological blockade and gene knockout mice. MATERIAL AND METHODS: Hearts (n = 6-8 per group) from wild type or homozygous B1 receptor gene knockout mice were isolated and perfused using the Langendorff technique. After a 30-min stabilisation period, the left coronary artery was occluded for 30 min followed by 60 min of reperfusion. In two separate groups of wild type hearts, B1 and B2 receptors were blocked with 3 nM of (des Arg9, Leu8)-bradykinin and 10 nM of Hoe 140, respectively, (started 15 min before ischemia and stopped before the reperfusion). RESULTS: Infarct size to risk zone (I/R) ratio was significantly reduced in hearts of knockout mice (11.3 +/- 2.1%) compared to those of wild type mice (25.7 +/- 1.7%). Furthermore, in wild type mice, I/R was significantly reduced in hearts perfused with the B1 receptor antagonist (12.8 +/- 2.4%) but not in hearts perfused with the B2 receptor antagonist (36.3 +/- 4.4%) compared to untreated hearts. Finally, a RT-PCR technique showed an activation of kinin B1 receptor gene transcription, in wild type hearts, subjected to the ischemia-reperfusion sequence. CONCLUSION: This study demonstrates that B1 receptors are induced during myocardial ischemia where they could play a detrimental role in mice. PMID- 12095174 TI - Protection by methyl inosine monophosphate (MIMP) against aerosol influenza virus infection in mice. AB - Methyl inosine monophosphate (MIMP) is a novel thymomimetic purine immunomodulator capable of enhancing a wide variety of immune responses. Intravenous (i.v.) administration of MIMP 1 day prior to influenza virus infection could confer partial protection and significantly increase the mean survival of treated mice. Intranasal (i.n.) administration of MIMP improved survival rates and incorporation of MIMP in squalane-saline emulsion 1 day prior to or 1 h after influenza infection conferred complete protection. Mice administered MIMP had reduced levels of lung virus and lower titers of circulating serum hemagglutination-inhibition (HI) antibodies. Complete survival and reduction of viral load after influenza challenge infection suggests effective stimulation by MIMP of protective responses against influenza virus. PMID- 12095173 TI - Estrogen deficiency results in enhanced expression of Smoothened of the Hedgehog signaling in the thymus and affects thymocyte development. AB - Aromatase is an essential enzyme for estrogen synthesis. We investigated the role of estrogen in thymocyte development using aromatase-deficient (ArKO) mice. Like its role as a regulator of bone metabolism through regulating osteoprotegerin (OPG) production, estrogen is involved in the processes of thymocyte development although aromatase mRNA was not detectable in the thymus. Thymic regression and reduced cellularity were evident in ArKO mice. The major difficulties in thymocyte development of ArKO mice were observed during the CD44+ CD25- stage at the cortico-medullary junction and during the CD44- CD25- stage at the subcapsular region where the estrogen receptor was expressed in the stromal cells. The proportion of thymocytes during the CD44+ CD25- stage was reduced. The progression of CD44- CD25+ cells to the CD44- CD25- stage was accelerated in ArKO mice possibly due to insufficient osteoprotegerin production in estrogen deficiency. However, the expression of Smoothened of the Hedgehog signaling was enhanced in CD4- CD8- double negative cells. This enhancement may result in impaired progression of CD44- CD25- cells to the CD4+ CD8+ double positive stage and impaired proliferation of CD4+ CD8+ double positive cells since Smoothened (Smo) is known to arrest cells as non-proliferating cells. This could be the reason why the proportion of CD3+ TCRbeta(high) cells during the late phase of thymocyte maturation was reduced in ArKO mice. From these observations, we propose that estrogen supports thymocyte development and maturation at many stages through many regulatory pathways including the sonic hedgehog- and the osteoprotegerin ligand (OPGL)-mediated signaling. PMID- 12095175 TI - Synthesis of factor D by gastric cancer-derived cell lines. AB - Synthesis of complement components in vitro by four human gastric cancer-derived cell lines, MKN28, MKN74, MKN45 and KATO-III, was studied. When these cells were cultured for 3 days without addition of any stimulator, 0.94 +/- 0.49, 2.10 +/- 0.59, 7.29 +/- 5.94 and 2.47+/- 1.34 ng of factor D/10(6) cells were detected in supernatants of MKN28, MKN74, MKN45 and KATO-III, respectively. Factor D production by these cells was reversibly inhibited by the presence of cycloheximide. Factors B, C3 and C2 were also detected in protein-free culture medium of these cell lines. Addition of tumour necrosis factor (TNF) to culture enhanced C3 and factor B secretion but depressed C2 secretion, without any distinct effect on factor D secretion. Since all cell lines tested secreted significant amounts of factor D without addition of any stimulator in medium, it is possible that factor D may be synthesized by gastric epithelial cells physiologically and constitutively. From a quantitative analysis of factor D secretion by these cells, factor D secreted by gastric tissue is likely to contribute to the factor D level in circulating blood. The possible mechanism of participation of complement system in inflammation of gastric epithelium was proposed. Thus, the present study may be significant for clarification of the mode of extrahepatic complement synthesis participating in mucosal immunity. PMID- 12095176 TI - Effect of (-)-epigallocatechin-3-gallate on respiratory burst of rat macrophages. AB - The toxic effects derived from overproduction of oxygen radicals [reactive oxygen species (ROS)] by immune cells can be partially abolished by the antioxidant activities of plant polyphenols. In the present study, we investigated the antioxidant action of a catechin, (-)-epigallocatechin-3-gallate (EGCG), on the respiratory-burst responses of rat peritoneal macrophages. EGCG at concentrations of 50-200 microM blocked the production of nitric oxide by macrophages stimulated in vivo with sodium thioglycollate then 5 days later in vitro with lipopolysaccharide and gamma-interferon. At 1-100 microM, EGCG also inhibited the extracellular liberation of oxygen radicals by resident peritoneal macrophages stimulated with the protein kinase C activator phorbol 12-myristate 13-acetate (PMA). At low concentrations (1-5 microM), EGCG increased the reduction of nitro blue tetrazolium (NBT) by the superoxide anions generated in the non-enzymatic system NADH/PMS, acting as a pro-oxidant agent, while at concentrations above 10 microM, EGCG acts as a scavenger of superoxide anions. These results show that EGCG is capable of modulating ROS production during the respiratory burst of rat peritoneal macrophages by acting as a superoxide anion scavenger. EGCG may therefore be useful in the prevention and treatment of diseases due to increased free radical production. PMID- 12095177 TI - Allergic bronchopulmonary aspergillosis: a review of a disease with a worldwide distribution. AB - Allergic bronchopulmonary aspergillosis (ABPA), an immunologically mediated lung disease, occurs predominantly in patients with asthma. This chronic relapsing disorder ranges clinically from mild asthma to fatal destructive lung disease and is caused by hypersensitivity to colonized Aspergillus fumigatus (Af). The immunopathogenesis of the disease is yet to be understood clearly. Specific IgE Af and IgG-Af, the serological markers, contribute to the diagnosis. Radiologically, ABPA is characterized by fleeting pulmonary infiltrates often confused with pulmonary tuberculosis. However, central bronchiectasis on computed tomography is considered to be the hallmark of the disease. Early diagnosis and therapy with prednisolone, the cornerstone of management, could alter the course of the disease and prevent the development of end-stage lung fibrosis. PMID- 12095178 TI - Antitussive effect of the leukotriene receptor antagonist zafirlukast in subjects with cough-variant asthma. AB - Cough-variant asthma (CVA) occurs in a subgroup of asthmatics whose sole or predominant respiratory symptom is cough. Although bronchodilators are often sufficient to treat CVA, refractory cough may require therapy with inhaled or systemic corticosteroids. In a randomized, double-blind, placebo-controlled, crossover study, we examined the effect of a 14-day course of the leukotriene receptor antagonist zafirlukast on subjective cough score and cough-reflex sensitivity to inhaled capsaicin in eight subjects with CVA refractory to inhaled beta agonists, and in five subjects refractory to inhaled corticosteroids. Seven of eight subjects experienced significant subjective and objective improvement in cough after treatment with zafirlukast. Mean (+/- SEM) cough score improved from 7.75 +/- 0.56 to 3.25 +/- 0.84 (p = 0.0006). Cough sensitivity to capsaicin was suppressed by zafirlukast in all subjects. Patients with CVA may represent a distinct subgroup of asthmatics whose afferent cough receptors within the respiratory epithelium are hypersensitive relative to those of patients with the typical form of asthma. Zafirlukast appears to be particularly effective in treating CVA by inhibiting the sensitivity of these receptors. Leukotriene receptor antagonists may offer an alternative to corticosteroids for the treatment of CVA refractory to inhaled bronchodilators. PMID- 12095179 TI - Reliability and validity of a system for coding asthma outcomes from medical records. AB - To evaluate the reliability and validity of a standardized asthma outcome coding system, we obtained medical records for 182 asthmatic children. Records were coded by trained staff using explicit and detailed criteria. Outcome variables coded included number of corticosteroid bursts, asthma-related physician contacts, emergency room visits, hospitalizations, and number of asthma episodes. Interrater reliability was excellent. Patterns of associations between the coded variables and other independently obtained outcome measures supported concurrent and construct validity. Given the intense scrutiny of health outcomes in the current managed-care marketplace, use of this system may foster further clinical research examining asthma outcomes. PMID- 12095180 TI - Relationship of wheezing to airflow obstruction in asthmatic children and a history of cough-variant asthma. AB - The aim of this study was to examine the relationship of wheezing to airflow obstruction during acute episodes of asthma in patients who had CVA (Cough variant asthma). Two groups of asthmatic children, one group with a past history of CVA (n = 13) and the other group without such a history (n = 14), were followed longitudinally for 12 months. During that time, they were evaluated for the presence of wheezing and the severity of airflow obstruction during acute episodes of asthma. Significant airflow obstruction occurred free of wheezing more frequently and the presence of clinical wheezing was associated with more severe airflow limitation, in asthmatic patients with a past history of CVA than in those without such a history. We conclude that asthmatic patients who have experienced CVA develop the wheezing symptom at a higher level of airflow obstruction. PMID- 12095181 TI - Serum B12 tryptase level as a marker of allergic airway inflammation in asthma. AB - Tryptase is a specific marker of mast-cell activation and plays a part in the pathophysiology of various allergic diseases including asthma, but little is known of the spillover of this enzyme into the systemic circulation. Therefore, we measured serum levels of mast-cell-derived tryptase in 21 patients with mild to moderate asthma and 20 healthy, subjects, using a B12 monoclonal antibody based immunofluoroassay that detects both monomers and tetramers of alpha- and beta-tryptases. There was a good correlation between serum and sputum tryptase levels, and, compared with healthy subjects (1.68 +/- 0.31 ng/ml), asthma patients had higher concentrations of serum tryptase (atopic asthma, 4.18 +/- 0.95 ng/ml, p = 0.022; nonatopic asthma, 3.93 +/- 0.82 ng/ml, p = 0.031). Although serum tryptase levels did not correlate with asthma symptom scores, peak expiratory flow, or forced expiratory volume in 1 s, they positively correlated with mast-cell and eosinophil counts (p = 0.041 and p = 0.025, respectively) and eosinophil cationic protein contents (p = 0.029) in induced sputum. These results suggest that serum tryptase detected with B12 antibody is a marker of allergic airway inflammation in asthma. PMID- 12095182 TI - Inhalant allergens as risk factors for the development and severity of mild-to moderate asthma in Hong Kong Chinese children. AB - Sensitization to aeroallergens has been found to be a dominant risk factor for asthma in population-based studies. Similar results in asthmatic children being managed in hospital-affiliated outpatient clinic have not been established. We therefore conducted a case-control study on asthmatic children attending a university hospital-based outpatient clinic to investigate the pattern of aeroallergen sensitization and to assess the correlation between asthma control and the presence of allergen-specific IgE-s. Asthmatic patients underwent a questionnaire assessment of their asthma control, skin prick tests (SPT) for allergen sensitization, and spirometric evaluation. Peripheral blood was collected from all subjects for in vitro serum specific IgE assay (RAST) to common aeroallergens. One hundred and seventy asthmatics (aged 9.8 +/- 3.7 years) and 57 age-matched control subjects were enrolled. Our patients had a median of two asthmatic attacks within 6 months prior to evaluation, and their median Disease Severity Score was 13. The median FEV1 in our asthmatic children was 98%, whereas serum logarithmic total IgE concentrations in patients and controls were 2.57 and 2.09, respectively (p < 0.0001). More than 85% of our asthmatic children were sensitized to house-dust mite (HDM), and sensitization to HDM and cat, as well as elevated serum total IgE concentration, was a significant risk factor for the development of asthma in this cohort. Several objective measures of asthma severity, as well as FEV1, correlated significantly with sensitization to HDM, pets, and cockroaches. In conclusion, indoor aeroallergens are one of the risk factors for the development and severity of mild-to-moderate asthma in Chinese children in Hong Kong. PMID- 12095183 TI - Isolated chronic cough with sputum eosinophilia caused by Humicola fuscoatra antigen: the importance of environmental survey for fungus as an etiologic agent. AB - We report here a 35-year-old man with isolated chronic cough associated with sputum eosinophilia in which Humicola fuscoatra (H. fuscoatra) antigen was an etiologic agent. He was admitted for the diagnosis and the treatment of his severe nonproductive cough. Although 80% of the nucleated cells in his induced sputum were eosinophils, he did not have bronchial hyperresponsiveness to methacholine or heightened bronchomotor tone. Bronchodilator therapy was not effective against his coughing. His cough worsened on his return home, suggesting the existence of some etiologic agent in his house. H. fuscoatra was isolated from his house, and the bronchoprovocation test with H. fuscoatra antigen was positive: i.e., development of coughing and decrease in capsaicin cough threshold (capsaicin concentration causing five or more coughs) from the prechallenge value of 31.3 microM to 1.95 microM at 6 and 48 hr, respectively, after the challenge. In addition, repeated environmental survey for fungi was suggestive of the importance of H. fuscoatra in the sputum eosinophilia. This is the first report concerning chronic cough with sputum eosinophilia caused by allergic reaction to H. fuscoatra antigen. PMID- 12095184 TI - Mouthpiece versus facemask for delivery of nebulized salbutamol in exacerbated childhood asthma. AB - We compared the bronchodilator response to salbutamol (albuterol) delivered by a compressed air nebulizer through a mouthpiece and via a facemask in 18 asthmatic children, to determine the most appropriate delivery method. Patients using a mouthpiece had significantly better mean percent increases in forced expiratory volume in 1 sec (FEV1) and in forced vital capacity (FVC) than those using a facemask 30 min after inhalation (FEV1, 56.4 +/- 32.6% vs. 28.9 +/- 19.1%, FVC: 34.4 +/- 26.4% vs. 7.5 +/- 14.9%, respectively). Nebulized therapy plays an important role in the management of bronchial asthma in children and should be delivered by a mouthpiece whenever possible in cases of exacerbated asthma. PMID- 12095185 TI - Towards excellence in asthma management (TEAM): a populational disease-management model. AB - Asthma management is not always optimal, and deficiencies such as inadequate treatment and insufficient patient education are often reported. Towards Excellence in Asthma Management (TEAM) is a four-phase disease management program of the Quebec Asthma Education Network (QAEN), to be carried out over a 5-year period. The program aims to achieve a continuous improvement of asthma management by caregivers and patients. The first phase, completed in January 2000, consisted of determining the actual level of asthma-associated morbidity and mortality in various Quebec regions. The second phase, which began in September 1999, included three parts: 1. Definition of the burden of asthma, taking into account the socioeconomic consequences of the disease and the quality of life of the patients, 2. Comparison of current medical practices with the Canadian Asthma Consensus Guidelines for adult and pediatric populations, 3. Evaluation of the level of compliance with medical treatment and with the environmental changes recommended to asthmatic patients. This phase is carried out via a cohort study of physicians, mainly general practitioners and pediatricians, generating a patient cohort study, in addition to substudies evaluating specific aspects of asthma care. Once the care gap is identified, it will be possible to define, apply, and evaluate a series of interventions for physicians, other health professionals, and patients. The interventions will be particularly targeted at regions where asthma incidence and morbidity are higher. We hope that this model of disease management will progressively reduce the burden associated with asthma, and potentially other chronic diseases, and will result in the more effective use of health services. PMID- 12095187 TI - Decomposing global self-esteem. AB - We argue in this paper for distinguishing two dimensions of global self-esteem, self-competence and self-liking. Studies 1 and 2 identify a corresponding pair of factors in Rosenberg's (1965) Self-Esteem Scale. Studies 3 and 4 examine the predictive value of the two-dimensional approach to self-esteem as reflected in the unique associations of self-competence and self-liking with negative life events and word recognition. PMID- 12095186 TI - Health-care resource use by asthmatics in Croatia. AB - We report a survey of the impact of asthma on daily life and on health-care resource use by adult asthma patients in four counties in Croatia. Over a 1-year period beginning November 1998, 185 general practitioners recruited 504 asthma patients, who completed a questionnaire about their drug therapy, health-care resource use, symptoms, limitations in daily activities, and work loss due to asthma. Over a 2-week period, inhaled corticosteroids were used by 51.5% of patients, short-acting and long-acting beta-agonists by 70.8% and 53.5%, respectively, and oral extended-release theophyllines by 29.8%. Nocturnal awakenings were experienced by 58.7% of patients, 75.5% experienced daytime symptoms, and 42.5% were limited in their daily activities. In the previous year, 12.5% of the patients were hospitalized, 33% missed work, and 70% reported reduced effectiveness in the workplace. In the previous 6 months, 35.2% of the patients visited an emergency room due to worsening of asthma symptoms. The human and socioeconomic burden of asthma in Croatia is considerable. These results can help guide the development of asthma policy in Croatia and raise awareness of asthma as a public health issue. PMID- 12095188 TI - Ego development and the ethics of care and justice: the relations among them revisited. AB - This study examined the links among ego development and the ethics of care and justice in 144 Norwegian men and women, 15 to 48 years old, taking into consideration age, sex, education, and verbal intelligence. As expected, the relationship between Loevinger's model of ego development and care-based moral reasoning as measured with Skoe's Ethic of Care Interview (ECI) was significantly stronger than the one between ego development and justice as measured with Rest's Defining Issues Test (DIT). Both ethics correlated significantly with verbal ability. Analyses showed that beyond its overlap with verbal intelligence, the variance shared between the ECI and ego development was substantial. By contrast, when verbal intelligence was controlled, the DIT was not significantly related to ego development or to the care ethic. PMID- 12095189 TI - When a grandiose self-image is threatened: narcissism and self-concept clarity as predictors of negative emotions and aggression following ego-threat. AB - Two studies examined the relation between narcissism, self-concept clarity, negative emotions, and aggression based on theoretical assumptions proposed by Baumeister, Smart, and Boden (1996). Narcissism and self-concept clarity were examined as predictors for anger, depression, and verbal aggression following ego threat, which was operationalized by a bogus performance feedback on an intelligence test. The second study also examined the mediating effects of participants' negative emotions to provide an additional explanation for the aggressive reactions after failure. As expected, narcissism and self-concept clarity were significant predictors of negative emotions and aggression after failure. In accordance with our hypothesis, high narcissists with low self concept clarity reacted with anger and aggression after failure, whereas less narcissistic individuals with high self-concept clarity showed feelings of depression and no aggression. The results also indicated that aggression was always directed toward the source of the ego-threatening feedback. Additionally, anger and depression could predict the aggressive response after failure but they did not mediate the relation between narcissism, self-concept clarity, performance feedback, and aggression. PMID- 12095190 TI - Delay of gratification: impulsive choices and problem behaviors in early and late adolescence. AB - High school students were offered a monetary incentive for participating in research. They were given a choice between a smaller fee immediately or a larger fee one week later. Compared to students who delayed gratification, those who chose the immediate fee showed more self-regulatory deficits. They showed greater involvement with cigarettes, alcohol, and marijuana, had a poorer self-concept and underperformed academically. A replication study with middle-school students and different reward parameters yielded equivalent results. Younger adolescents who chose the immediate monetary incentive showed a similar pattern of problem behaviors as the high school students. The findings indicate that this simple choice-delay procedure yields an unobtrusive behavioral measure of self regulation and offers a developmentally appropriate extension of the delay-of gratification paradigm for use with older children and adolescents. PMID- 12095191 TI - Inhibited power motivation and persuasive communication: a lens model analysis. AB - The authors tested the hypothesis that after motive arousal, individuals with an inhibited power motive (IPM) would excel at a persuasive task and explored the behavioral strategies IPM individuals use to that purpose. Sixty-eight participants presented their point of view on a controversial subject to another person. Power motivation and inhibition were both assessed by a picture-story test. Prior to their presentation, half of the participants imaginatively explored the ensuing task. The other half was assigned to a no-imagery control condition. Lens model analysis of videotaped presentations revealed that IPM participants in the imagery condition were judged to be the most persuasive of all participants. This interactive effect of power motivation, inhibition, and imagery condition was accounted for by three behavioral cues: verbal fluency, gesturing, and eyebrow lifts. No comparable effects emerged among no-imagery participants. PMID- 12095192 TI - Effects of rod activity on color perception with light adaptation. AB - To investigate the effect of rod activity on color perception with light adaptation, chromaticity shifts of monochromatic test lights were measured as a function of background field intensity at 17 deg in the nasal field of view. The measurements were performed both after complete dark adaptation and during the cone-plateau period at a mesopic test intensity level of 15 photopic trolands. To clarify the mechanisms underlying the chromaticity shifts obtained, six supplementary experiments were performed. The results of the experiments strongly suggest that at scotopic background intensities, light adaptation of rods, both within and adjacent to the test area, may reduce rod signals triggered by the test light and thereby produce marked chromaticity shifts with light adaptation. At mesopic background intensities, cones in the background field become activated and may influence the chromaticity shift with light adaptation both by suppressing signals from rods elicited by the test light and by producing a selective chromatic adaptation. PMID- 12095193 TI - Contrast discrimination with pulse trains in pink noise. AB - Detection performance was measured with sinusoidal and pulse-train gratings. Although the 2.09-cycles-per-degree pulse-train, or line, grating contained at least eight harmonics all at equal contrast, it was no more detectable than its most detectable component. The addition of broadband pink noise designed to equalize the detectability of the components of the pulse train made the pulse train approximately a factor of 4 more detectable than any of its components. However, in contrast-discrimination experiments, with a pedestal or masking grating of the same form and phase as the signal and with 15% contrast, the noise did not affect the discrimination performance of the pulse train relative to that obtained with its sinusoidal components. We discuss the implications of these observations for models of early vision, in particular the implications for possible sources of internal noise. PMID- 12095194 TI - Contrast discrimination with sinusoidal gratings of different spatial frequency. AB - The detectability of contrast increments was measured as a function of the contrast of a masking or "pedestal" grating at a number of different spatial frequencies ranging from 2 to 16 cycles per degree of visual angle. The pedestal grating always had the same orientation, spatial frequency, and phase as the signal. The shape of the contrast-increment threshold versus pedestal contrast (TvC) functions depends on the performance level used to define the "threshold," but when both axes are normalized by the contrast corresponding to 75% correct detection at each frequency, the TvC functions at a given performance level are identical. Confidence intervals on the slope of the rising part of the TvC functions are so wide that it is not possible with our data to reject Weber's law. PMID- 12095195 TI - Tip-tilt error for extremely large segmented telescopes: detailed theoretical point-spread-function analysis and numerical simulation results. AB - We present an analysis of point-spread functions for segmented mirrors affected by random tip-tilt errors on each segment. In addition to Strehl ratio evaluation, this analysis considers key characteristics such as the intensity and the location of speckles and secondary peaks and the relative energy distribution between these features. We develop a method to describe the shape of a nonaveraged point-spread function and deduce the final expressions for ensemble averaged characteristics. Based on Keck-type hexagonal segmentation geometry, our study is extended to an arbitrary number of segments, and we describe qualitatively the transition from the case of a mirror with few segments to that of a mirror with several hundred segments--prototype of the next generation of Extremely Large Telescopes. PMID- 12095196 TI - Statistical-information-based performance criteria for Richardson-Lucy image deblurring. AB - Iterative image deconvolution algorithms generally lack objective criteria for deciding when to terminate iterations, often relying on ad hoc metrics for determining optimal performance. A statistical-information-based analysis of the popular Richardson-Lucy iterative deblurring algorithm is presented after clarification of the detailed nature of noise amplification and resolution recovery as the algorithm iterates. Monitoring the information content of the reconstructed image furnishes an alternative criterion for assessing and stopping such an iterative algorithm. It is straightforward to implement prior knowledge and other conditioning tools in this statistical approach. PMID- 12095197 TI - Quantitative assessment of image quality enhancement due to unsharp-mask processing in x-ray fluoroscopy. AB - Spatial unsharp-mask processing and its variants are commonly used in x-ray radiography to enhance image contrast. We investigated the effect of three unsharp-masking filter kernels of different sizes on the detection of an advanced guidewire tip in simulated x-ray fluoroscopy image sequences. To isolate the effect of visual temporal processing, we repeated the experiments on single images. Filter gains were selected so that all three kernels increased the contrast of a 0.018-in. (0.457-mm) guidewire by a factor of 2 but had different effects on image noise and signal profiles. There was no statistically significant effect of unsharp masking on human-observer performance in single images. However, all three kernels significantly improved average performance in image sequences, and the guidewire contrast required for detection was reduced by 32%-40%. A prewhitening channelized observer model predicted the disparity between sequences and single images and fitted measurements at different kernel sizes well. A nonprewhitening observer model did not. We conclude that unsharp masking is a simple and effective method of improving guidewire visualization in fluoroscopically guided interventional procedures and that quantitative image quality studies are essential for evaluation of image-processing techniques in sequences such as x-ray fluoroscopy. PMID- 12095198 TI - Imaging perfectly conducting objects as support of induced currents: Kirchhoff approximation and frequency diversity. AB - The problem of determining the shape of perfectly conducting objects from knowledge of the scattered electric field is considered. The formulation of the problem accommodates the nature of the distribution of the induced surface current density. Thus, as the unknown representing the object's contour, a single layer distribution is chosen so that the contour of the scatterer is described by its support. The nonlinear unknown-data mapping is then linearized by means of the Kirchhoff approximation, and the problem is recast as the inversion of a linear operator acting on a distribution space. An extension of the singular value decomposition approach to solve the linearized problem is provided and numerical results are presented. PMID- 12095199 TI - Discrete Green's methods and their application to two-dimensional phase unwrapping. AB - A fully self-contained discrete framework with discrete equivalents of Stokes's, Gauss's, and Green's theorems is presented. The formulation is analogous to that of continuous operators, but totally discrete in nature, and the exact relationships derived are shown to hold provided that a set of predefined rules is followed in building discrete contours and domains. The method allows for an analytical rigor that is not guaranteed if one translates the classical continuous formulations onto a discretized approximated framework. We clarify several issues related to the use of discrete operators, which may play a crucial role in specific applications such as the two-dimensional phase-unwrapping problem, chosen as our main application example, and we show that reconstruction on irregular domains and/or in the presence of undersampling and noise is better formulated in the discrete framework than in the continuous domain. PMID- 12095200 TI - Phase retrieval, error reduction algorithm, and Fienup variants: a view from convex optimization. AB - The phase retrieval problem is of paramount importance in various areas of applied physics and engineering. The state of the art for solving this problem in two dimensions relies heavily on the pioneering work of Gerchberg, Saxton, and Fienup. Despite the widespread use of the algorithms proposed by these three researchers, current mathematical theory cannot explain their remarkable success. Nevertheless, great insight can be gained into the behavior, the shortcomings, and the performance of these algorithms from their possible counterparts in convex optimization theory. An important step in this direction was made two decades ago when the error reduction algorithm was identified as a nonconvex alternating projection algorithm. Our purpose is to formulate the phase retrieval problem with mathematical care and to establish new connections between well established numerical phase retrieval schemes and classical convex optimization methods. Specifically, it is shown that Fienup's basic input-output algorithm corresponds to Dykstra's algorithm and that Fienup's hybrid input-output algorithm can be viewed as an instance of the Douglas-Rachford algorithm. We provide a theoretical framework to better understand and, potentially, to improve existing phase recovery algorithms. PMID- 12095201 TI - Guided-mode resonant grating filter with an antireflection structured surface. AB - We describe a new structure of guided-mode resonant grating (GMRG) filters with low sideband reflectance. This GMRG filter consists of a high-index thin film on an antireflective structured surface called "moth-eye structure." Since the high index film undulates along the surface structure, the film acts as a modulated optical waveguide. An incident light wave satisfying a resonant condition is reflected by the GMRG filter, and nonresonant light waves pass through the filter. This GMRG filter is valid for reducing reflection of nonresonant light waves in a wide spectral range. The resonant reflection of this new filter was investigated by numerical calculation based on an electromagnetic grating analysis. In the case of a triangular antireflective surface structure whose thickness is 2x greater than its period, the sideband reflectance for nonresonant light waves was lower than 0.5% for TM-polarized light in a wide range of wavelengths. PMID- 12095202 TI - Metallic inductive and capacitive grids: theory and experiment. AB - We present theoretical modeling and experimental validation of both capacitive (dot) and inductive (hole) metallic crossed gratings in the mid-infrared (2-5 microm). The gratings are fabricated by use of interferometric lithography and modeled by use of rigorous coupled-wave analysis. Our experimental and numerical investigations of the transmittance spectra of these gratings suggest that, as in inductive grids, the behavior of capacitive grids is described by the coupling of the incident light into surface plasma waves. PMID- 12095203 TI - Self-imaging properties of polychromatic Gaussian beams: application to wavelength demultiplexing and space routing. AB - We propose an analytical model of the self-imaging of Gaussian beams and discuss the impact of the finite aperture and the beam truncation on the quality of the self-image reconstruction. Extension to polychromatic operation is then presented in the context of wavelength division multiplexing systems. From the first point we derive conditions for good self-imaging, compatible with the requirements of the telecommunication environment. A basic optical setup is then proposed to implement both wavelength demultiplexing and routing in a lensless configuration. PMID- 12095204 TI - Generalized beam-propagation factor of partially coherent beams propagating through hard-edged apertures. AB - The second-order intensity moments and beam-propagation factor (M2 factor) of partially coherent beams have been generalized to include the case of hard-edged diffraction. A laser beam with amplitude modulation and phase fluctuation and a Gaussian Schell-model beam are taken as two typical examples of partially coherent beams. Analytical expressions for the generalized M2 factor are derived. PMID- 12095205 TI - Analyzing the power coupled between partially coherent waveguide fields in different states of coherence. AB - A procedure is described for calculating the power coupled between partially coherent waveguide fields that are in different states of coherence. The method becomes important when it is necessary to calculate the power transferred from a distributed source S to a distributed load L through a length of multimode metallic, or dielectric, waveguide. It is shown that if the correlations between the transverse components of the electric and magnetic fields of S and L are described by coherence matrices M and M', respectively, then the normalized average power coupled between them is (eta) = Tr[MM']/Tr[M]Tr[M'], where Tr denotes the trace. When the modal impedances are equal, this expression for the coupled power reduces to an equation derived in a previous paper [J. Opt. Soc. Am. A 18, 3061 (2001)], by use of thermodynamic arguments, for the power coupled between partially coherent free-space beams. PMID- 12095206 TI - Photon path distribution in inhomogeneous turbid media: theoretical analysis and a method of calculation. AB - The photon path distribution (PPD) is a measure that I have developed to express optical responses in inhomogeneous turbid media in the time and frequency domains. The PPD is defined by local photon pathlengths of possible photons having total zigzag pathlengths I between the points of light input and detection. Such a distribution is independent of absorption and is uniquely determined for the medium under quantification. I show that the PPD is derived through the local photon count of the possible photons arising from an optical impulse incident on an imaginary medium having the same optical properties as the medium under quantification, except for the absence of absorption. The formulas derived can be used to calculate the PPD simultaneously with, for example, the numerical calculation of a diffusion equation. PMID- 12095207 TI - Extended analysis of curvature sensing. AB - Curvature sensors are used in adaptive optics to measure the wave-front aberrations. In practice, their performance is limited by their nonlinear behavior, which we characterize by solving simultaneously the irradiance transport equation and the accompanying wave-front transport equation. We show how the presence of nonlinear geometric terms limits the accuracy of the sensor and how diffraction effects limit the spatial resolution. The effect of photon noise on the sensor is also quantified. PMID- 12095208 TI - Method for deriving the complete solution set for three-mirror anastigmatic telescopes with two spherical mirrors. AB - The "plate-diagram" method of quantifying and manipulating the Seidel aberrations of an optical system has been used to develop a procedure that has successfully determined the complete solution set of three-mirror anastigmats in which two surfaces are left strictly spherical. The procedure also readily identified solutions in which the Petzval sum is zero, and four distinct families of flat field three-mirror anastigmats with two mirrors strictly spherical have thus been found. The success of the method is strong support for the argument that algebraic approaches to optical design can yield results distinctly superior to currently favored optimization-based design methods, at least for some types of optical systems. PMID- 12095209 TI - Effects of source coherence and aperture array geometry on optical sectioning strength in direct-view microscopy. AB - Laser sources offer a possible solution to the problem of low light throughput in direct-view microscopes (DVMs). However, coherent source DVMs have been shown to suffer from problems such as increased sidelobes in the depth response because of coherent cross talk between neighboring apertures. We explore theoretically how source coherence affects the depth responses of DVMs by employing various aperture spacings and number of apertures. We show that, contrary to expectation, closely spaced apertures can result in decreased full width at half-maximum of the depth response curve. We explain this as an effect of destructive interference when cross talk between neighboring apertures occurs. Using apertures arranged in a square grid as an example, we move on to show that the use of aperture arrays that consist of regularly arranged apertures can accentuate the problematic sidelobes of the depth response. We show that arranging pinholes in a rectangular grid rather than a square grid can improve the optical sectioning strength significantly. Finally, by examination of the depth responses corresponding to the infinite-pinhole-array limit, we make some general statements about source coherence and the characteristics of arrays that are likely to perform well. PMID- 12095210 TI - Analysis of accuracy of measurement of quadratic electro-optic coefficients in uniaxial crystals: a case study of KDP. AB - Taking KDP as an example of a uniaxial crystal, we analyze contributions to its quadratic electro-optic response with a view to explaining differences in published values of certain quadratic coefficients. In an eigenvalue theory of light propagation we show that the linear electro-optic coefficient contributes to the quadratic response, even under ideal laboratory conditions. In addition, the effect of imperfect crystal cutting and alignment is investigated by means of computer calculations based on the Jones calculus. It is found that, for relatively small inaccuracies, the calculated values of the quadratic coefficient g(xyxy) are approximately two orders of magnitude greater when measured with a static field than with a dynamic one. This finding could explain the observed spread in some results for KDP-type crystals. PMID- 12095211 TI - Nonparaxial description of reflection and transmission at the interface between an isotropic medium and a uniaxial crystal. AB - Angular spectra of reflected and transmitted fields, induced by an arbitrary electromagnetic beam passing through the planar interface between a homogeneous medium and a uniaxially anisotropic medium, are derived and related to the incident medium. By using these formulas, we obtain the expressions for paraxial and slightly nonparaxial fields. The reflected paraxial field is related to the incident one by means of Fresnel relations; the transmitted paraxial field is the superposition of an ordinary and an extraordinary beam, multiplied by the Fresnel coefficient. We find that the nonparaxial corrections, owing to the medium discontinuity, are larger than their free-propagation counterparts and that they are very simply related to the paraxial solutions of the incident beam. The case of two homogeneous media with different refractive indices is also discussed. The general expressions obtained are applied to the case of a nonparaxial Gaussian beam. PMID- 12095212 TI - Terahertz multistatic reflection imaging. AB - We describe a new imaging method using single-cycle pulses of terahertz (THz) radiation. This technique emulates the data collection and image processing procedures developed for geophysical prospecting and is made possible by the availability of fiber-coupled THz receiver antennas. We use a migration procedure to solve the inverse problem; this permits us to reconstruct the location, the shape, and the refractive index of targets. We show examples for both metallic and dielectric model targets, and we perform velocity analysis on dielectric targets to estimate the refractive indices of imaged components. These results broaden the capabilities of THz imaging systems and also demonstrate the viability of the THz system as a test bed for the exploration of new seismic processing methods. PMID- 12095213 TI - Convergence of electromagnetic field components across discontinuous permittivity profiles: comment. AB - The inverse rule that is described in a recent paper [J. Opt. Soc. Am. A 17, 491 (2000)] is not a multiplication rule for multiplying two infinite series, because it does not address how the terms of two series being multiplied are combined to form the product series. Furthermore, it is not the one that is being used in numerical practice. Therefore the insight that the paper provides into why the inverse rule yields correct results at the points of complementary discontinuities is questionable. PMID- 12095214 TI - Evanescent and propagating electromagnetic fields in scattering from point-dipole structures: comment. AB - I comment on the recent paper by Setala et al. [J. Opt. Soc. Am. A 18, 678 (2001)]. In that paper the calculation is entirely and exactly adopted from our previous work except the evanescent-field propagator. I shall demonstrate that the replacement of the propagator flaws the calculation. In their paper the numerical results show rich subwavelength information at absolute far-field distances (four times larger than the wavelength). I shall point out that the evanescent resolution at far-field distances cannot be of subwavelength. In the paper by Setala et al. the description of previous work is inaccurate and misleading. I shall give what I consider to be the correct guidelines for readers to learn about the standing controversy regarding the evanescent field of a dipole. PMID- 12095215 TI - Cell transplantation for Parkinson's disease. PMID- 12095216 TI - Intrathecal spinal progenitor cell transplantation for the treatment of neuropathic pain. AB - Injury to, or dysfunction of, the nervous system can lead to spontaneous pain, hyperalgesia, and/or allodynia. It is believed that the number and activity of GABAergic neurons gradually decreases over the dorsal horn. Glutamic acid decarboxylase (GAD) immunocompetence has been demonstrated on spinal progenitor cells (SPCs) cultivated in vitro. The intrathecal implantation of these cultivated progenitor cells may provide a means of alleviating neuropathic pain. Chronic constriction injury (CCI) of the sciatic nerve was used to induce chronic neuropathic pain in the hind paw of rats. SPCs (1 x 10(6)) were implanted intrathecally on the third day after the CCI surgery. The behavioral response to thermal hyperalgesia was observed and recorded during the 14 days postsurgery. Various techniques were utilized to trace the progenitor cells, confirm the differentiation, and identify the neurotransmitters involved. GAD immunoactivity was revealed for 65% of the cultivated spinal progenitor cells in our study. We also determined that transplanted cells could survive more than 3 weeks postintrathecal implantation. Significant reductions were demonstrated for responses to thermal stimuli for the CCI rats that had received intrathecal SPC transplantation. A novel intrathecal delivery with SPCs reduced CCI-induced neuropathic pain. PMID- 12095217 TI - Alternative sources of neurons and glia from somatic stem cells. AB - Stem cell populations have been shown to be extremely versatile: they can generate differentiated cells specific to the tissue in which they reside and descendents that are of different germ layer origin. This raises the possibility of obtaining neuronal cells from new biological source of the same adult human subjects. In this study, we found that epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) cooperated to induce the proliferation, self renewal, and expansion of neural stem cell-like population isolated from several newborn and adult mouse tissues: muscle and hematopoietic tissues. This population, in both primary culture and secondary expanded clones, formed spheres of undifferentiated cells that were induced to differentiate into neurons, astrocytes, and oligodendrocytes. Brain engraftment of the somatic-derived neural stem cells generated neuronal phenotypes, demonstrating the great plasticity of these cells with potential clinical application. PMID- 12095218 TI - Injection of chemotherapeutic microspheres and glioma. III: Parameters to optimize efficacy. AB - Injectable microspheres may provide a means of providing local, sustained exposure of glioma to chemotherapeutics to improve patient survival. Using a rodent model of surgically resected glioma, we previously demonstrated that direct injections of chemotherapeutic microspheres into the tissue surrounding a resection cavity provide superior survival effects over injections of the same microspheres directly into the surgical cavity. The present experiments extended this novel observation by exploring several parameters related to the use of intraparenchymal injections of chemotherapeutic microspheres to treat glioma. Using a rat model of resected glioma, several principles regarding the use of local sustained release carboplatin microspheres were established. First, an inverted U dose-response was observed, wherein further dose escalation beyond the optimal dose was not efficacious and indeed produced significant local toxicity. Second, it was necessary to expose approximately 40% of the tumor margin to sustained release carboplatin in order to increase survival in this model. Survival was not enhanced when the proportion of the tumor margin exposed to carboplatin was only 20%. Third, the distribution of the chemotherapeutic microsphere injections along the tumor perimeter was shown to be important, requiring that the entire perimeter be proportionately exposed to the chemotherapeutic agent. Together, these data continue to support the development of chemotherapeutic microspheres for treating glioma. However, they also caution that a number of fundamental parameters can profoundly influence the efficacy that might be expected from local sustained delivery. Careful attention to these principles is not only required if chemotherapeutic microspheres are to be used efficaciously, but these principles should provide a foundation to further optimize the potential of this and other polymeric delivery systems under development for local, intraparenchymal drug delivery to glioma. PMID- 12095219 TI - Injection of chemotherapeutic microspheres and glioma. IV: Eradicating tumors in rats. AB - Polymer microspheres can be easily injected into the brain to provide a local and sustained delivery of chemotherapeutics to a tumor or surrounding tissue subject to high rates of tumor recurrence following surgery. Building on previous studies that established the clear advantage of local, peritumoral injections of sustained release microspheres, the following experiments utilized two different approaches for maximizing the survival benefit in glioma-bearing rats. In the first experiment, a previously grown cortical tumor was debulked and animals received either one or two treatments with carboplatin-loaded microspheres (either 200 or 800 microg total carboplatin per treatment). In each case, the microspheres were injected along the perimeter of the resection cavity with each treatment separated by 20 days. Survival studies clearly demonstrated that two, temporally spaced injections were superior to a single series of injections. At the lowest dose tested (200 microg), median survival was increased an additional 40% over that in animals receiving one treatment. At the higher dose (800 microg), one third of the animals receiving two separate treatments were long term survivors (> 150 days) and showed complete eradication of the tumor on histological examination. In the second experiment, we directly compared the efficacy produced by sustained release carboplatin or 1,3-bis[2-chloroethyl]-1 nitrourea (BCNU) alone versus injecting carboplatin and BCNU-loaded microspheres blended together as a single suspension. Carboplatin and BCNU both enhanced survival, with BCNU being significantly less effective than carboplatin. However, the greatest improvements in survival were seen when a blended suspension of carboplatin and BCNU microspheres was injected around the surgical cavity. In contrast, spatially alternating injections of BCNU and carboplatin microspheres was significantly less effective and the increase in survival was no greater than that achieved with BCNU alone. These data offer further support for the potential utility of local, sustained release chemotherapeutic microspheres for treating glioma. Moreover, they suggest that injectable chemotherapeutic microspheres may offer important advantages by (a) permitting multiple, temporally spaced injections to be made, as needed, and (b) providing the opportunity to deliver combinations of several different efficacious drugs directly to the tumor site to enhance survival beyond what can be achieved with delivery of any single chemotherapeutic agent. PMID- 12095221 TI - Long-term insulin independence following repeated islet transplantation in totally pancreatectomized diabetic pigs. AB - Clinical islet transplantation (Tx) in type I diabetic patients has been successful so far only in a minority of cases, probably because of multiple factors, partly immunologic and partly nonimmunologic in nature. Preclinical studies of islet Tx in large animals are still needed to clarify the reasons and find possible solutions. In this study, we tested the feasibility of noninvasive, repeated intrahepatic allo-Tx of porcine pancreatic islets obtained from multiple donors, in pigs rendered diabetic by total pancreatectomy (Pct). In group I Yucatan miniature swine (n = 6), after induction of diabetes by Pct, repeated islet allo-Tx of > or = 80% pure islets was performed. Islets obtained from two pigs of the Hanford breed were injected twice a week, half freshly isolated and half 48-h cultured, over a period of 11 days, for a total of 23,647 +/- 1617 islet equivalents (IE)/kg recipient body weight (BW). In group II Yucatan miniature swine (n = 3), after Pct, a single allo-Tx of > or = 80% pure islets, previously obtained from two donors of the Hanford breed, was performed, using a total of 22,416 +/- 1124 IE/kg BW. In group III Yucatan miniature swine (n = 3), auto-Tx of 60-75% pure islets, averaging 2980 +/- 424 IE/kg BW, was performed a few hours after Pct. Group IV Yucatan mini pigs (n = 3) underwent Pct and were used as diabetic controls. Group V animals (n = 3) were normal control Yucatan mini pigs. Porcine islets were isolated by a modification of the standard collagenase digestion and Ficoll gradient purification method. Donors and recipients were chosen on the basis of moderate to high mutual alloreactivity in mixed lymphocyte culture (MLC). In groups I and II, cyclosporine A (CsA) was started 4 days before allo-Tx, at the dose of 15 mg/kg IM, and then gradually reduced to 4 mg/kg IM. In all group I animals, normal fasting blood glucose (FBG) was restored within 2-3 weeks. Two normoglycemic pigs died of acute pneumonia at 33 and 112 days, respectively, and one animal became progressively hyperglycemic at 100 days. After 3 months, discontinuation of CsA treatment resulted in FBG increase in two group I animals. In one pig, CsA was stopped after 151 days, and normoglycemia persisted until euthanasia, after 8 months. In group II pigs, normoglycemia lasted 4-20 days, with a progressive increase of insulin requirement thereafter. In group III animals, after islet auto-Tx, normoglycemia lasted 7-10 days, while insulin daily requirement progressively increased thereafter, stabilizing at 0.4 IU/kg/day, corresponding to about one third of the amount required in diabetic controls. The single most important result in this series of experiments is that intraportal allo-Tx of a sufficient islet mass, divided in multiple subtherapeutic doses, produced a better metabolic long-term control in comparison to a single injection of the same amount of islets. The technique of multiple-donor repeated islet Tx may prove useful to overcome the problem of primary nonfunction or early graft failure, currently limiting the success of clinical islet Tx in most cases. PMID- 12095220 TI - Xenografts of MHC-deficient mouse embryonic mesencephalon improve behavioral recovery in hemiparkinsonian rats. AB - The limited availability of human embryonic tissue for dopamine cell transplants in Parkinson's patients has led to an increased interest in using xenogeneic donor tissue. Unfortunately, without aggressive immunosuppression, such brain xenografts are rejected by the host immune system. Chronic brain xenograft rejection is largely mediated by helper T cells, which require presentation of xenoantigens by major histocompatability complex (MHC) class II for their activation. We examined survival and function of xenografts of E13 mouse mesencephalon deficient in either MHC class I, class II, or both after transplantation into adult hemiparkinsonian rats without immunosuppression. Recipients received grafts from C57BL/6 mice that were either: 1) wild-type (wt), 2) MHC class I knockout (KO), 3) MHC class II KO, 4) MHC class I and II double KO, or 5) saline sham transplants. At 6 weeks after transplantation, recipients of MHC class I KO, class II KO, and double KO xenografts significantly reduced methamphetamine-induced circling rate while rats with wt xenografts and sham operated rats showed no improvement. MHC class II KO grafts had the greatest number of surviving dopamine neurons. All transplants, including saline sham controls, contained infiltrating host MHC class II-positive cells. Saline sham grafts and MHC class II KO xenografts contained significantly fewer infiltrating host MHC class II-positive cells than did wt grafts. Our results show that MHC class II-deficient xenografts survive transplantation for at least 6 weeks in the absence of immunosuppression, reduce rotational asymmetry, and provoke lesser immune reaction than wt grafts. PMID- 12095222 TI - Newly designed compliant hierarchic hybrid vascular grafts wrapped with a microprocessed elastomeric film--I: Fabrication procedure and compliance matching. AB - The object of this study was to develop a compliant hybrid vascular graft minimally supported by an elastomeric scaffold for arterial replacement. The hybrid vascular grafts designed were composed of three layers: an inner surface lined with endothelial cells (ECs); a hybrid medial tissue composed of a collagenous gel embedded with smooth muscle cells (SMCs); and an outer layer made of a laser-processed micropored segmented polyurethane (SPU) film with the circular pore size (diameter 150 microm) but different film thickness (50-200 microm) and different pore-to-pore distances (1 or 4 mm). The approximate dimensions of the hybrid vascular graft without the SPU film were as follows: inner diameter, 5 mm; length, 5 cm; thickness, 50 microm. The intraluminal pressure-external diameter relationship was measured by infusion of a phosphate buffer solution into the hybrid vascular graft. Canine carotid arteries and commercially available ePTFE grafts served as controls. Decrease in the thickness of the SPU film and increase in the pore density of the SPU film increased the pressure-dependent distensibility of the hybrid vascular grafts. The thinner the film and higher the pore density, the more compliant was the hybrid graft. The pressure-induced distensibility of the designed hybrid graft was found to be close to that of native carotid arteries. PMID- 12095223 TI - Newly designed compliant hierarchic hybrid vascular graft wrapped with microprocessed elastomeric film--II: Morphogenesis and compliance change upon implantation. AB - Hierarchic structured hybrid tubular vascular media composed of endothelial cells (ECs), which covered the luminal surface, and smooth muscle cells (SMCs), which resided in the tubular collagen gel, were wrapped with thin segmented polyurethane elastomeric films designed to provide compliance matching with native arteries and transmural tissue permeability using a laser-directed ablation technique to provide different pore densities. Two hybrid grafts with high and low pore densities (inner diameter: 150 microm and length: 4 cm), and exhibiting pressure-dependent distensibility in response to pulsatile pressure, were bilaterally implanted into canine common arteries for up to 6 months. Irrespective of the pore density, high patency was achieved and no dilation and bursting occurred. Maintenance of full endothelialization during the entire course of implantation period was observed for the graft wrapped with the film with higher pore density. On the other hand, the graft wrapped with the film with lower pore density exhibited markedly reduced endothelialization at a later period of implantation, probably due to delamination of neoarterial tissue from the segmented polyurethane (SPU) surface. There were some differences in transmural tissue ingrowth between the two grafts. At anastomotic sites, neoarterial thickness for type A graft was smaller than that for type B graft regardless of the implantation period. Slightly reduced compliance was observed for both types of grafts at the sixth month of the implantation period. This study indicates that a hybrid vascular graft minimally supported with a thin elastomeric film can be used to replace diseased arteries if micropores are well designed for tissue permeability and anchoring. PMID- 12095224 TI - Involvement of glutathione and glutathione-related enzymes in the protection of normal and trisomic human fibroblasts against daunorubicin. AB - We measured the glutathione content, and the activity of glutathione-related enzymes and DT-diaphorase in cultured normal (cell line: S-126) and trisomic (cell lines: S-158, S-240) human fibroblasts exposed to daunorubicin (DNR). Determination of reduced and total glutathione levels, and measurement of the activity of glutathione peroxidase, glutathione reductase, glutathione-S transferase and DT-diaphorase were performed spectrophotometrically. Human fibroblasts were exposed to 4 microm DNR for 2 h, and the cells placed in drug free medium for 6, 12, 24, 48, and 72 h. Cellular levels of GSH and total glutathione decreased following exposure to DNR. However, the ratio of GSH to total glutathione returned to control levels only in trisomic cells. These changes were concomitant with increasing glutathione-S-transferase and glutathione reductase activities. DNR also significantly increased the activity of Se-independent peroxidase and DT-diaphorase in trisomic fibroblasts. Marked increases in the activity of Se-dependent peroxidase and DT-diaphorase alone were seen in normal cells. The results provide the first evidence that DNR can induce alterations in the level of glutathione and glutathione-dependent enzymes in trisomic fibroblasts as compared to normal cells, which may provide additional protection against daunorubicin-induced oxidative stress in trisomic fibroblasts. PMID- 12095225 TI - DNA plasmid transmission in yeast is associated with specific sub-nuclear localisation during cell division. AB - Circular plasmids in yeast carrying only an origin of DNA replication (ARS) exhibit maternal inheritance bias (MIB) and are poorly transmitted from mother to daughter cell during division. A variety of different sequences that overcome MIB have been described, including centromeric sequences (CEN), telomere-associated repeats, silencer sequences and a specific system encoded by the endogenous 2 micron circle plasmid requiring the cis-acting locus STB and the proteins Rep1 and Rep2. In each case, DNA segregation between mother and daughter cells is dependent on DNA-protein interactions. Using plasmids carrying multiple copies of a lac repressor binding sequence, we have localised DNA molecules in the yeast nucleus using a green fluorescent protein (GFP)-lac repressor fusion protein. We compared GFP localised plasmids carrying a centromere sequence with plasmids based on 2 micron circle carrying or lacking the STB sequences required for their segregation. We show that GFP localised plasmid carrying the complete STB locus co-localises with the plasmid proteins Rep1 and Rep2 to discrete chromatin sites. These sites are distinct from both the telomeres and from sites of cohesin binding. Deletion of the region of STB essential for the stability of the plasmid, leads to a loss of plasmid association with chromatin, relocalisation of plasmids towards the nuclear periphery, and a decrease in the Rep1 protein associated with the plasmid. We conclude that specific plasmid localisation is likely to be important in the overcoming of MIB in yeast. PMID- 12095226 TI - The effects of oxidative stress and altered intracellular calcium levels on vesicular transport of apoE-EGFP. AB - Apolipoprotein E (apoE) plays a role in the distribution of lipid within many organs and cell types in the human body, including the central nervous system (CNS). The apoE4 isoform is also an established risk factor for late-onset Alzheimer's disease (AD), however its role in the aetiology of the disease remains largely unknown. Therefore, as AD is a late-onset disease, we sort to investigate how conditions hypothesised to model ageing affect apoE metabolism, such as the transport of apoE along the secretory pathway. Two of these models include oxidative stress and calcium deregulation. Using apoE-EGFP-expressing astrocytoma cell lines we established that vesicle number and velocity are up regulated under oxidative stress conditions, and slowed under KCl induced calcium deregulation. Although these findings apply to cells in general under these two stress conditions, the up-regulation of apoE in particular may be a response to cell injury with implications for neurodegeneration such as that found with late onset AD. PMID- 12095228 TI - Effect of antisense hTERT mRNA oligodeoxynucleotide on telomerase activity of leukemic cells. AB - The effect of antisense hTERT mRNA oligodeoxynucleotide on telomerase activity of leukemia cells was explored and investigated in the present study. Telomerase activity was measured by the telomerase PCR ELISA assay kit (TRAP); hTERT mRNA expression by reverse transcription polymerase chain reaction (RT-PCR) assay and gel-image system, hTERT protein by immunochemistry and flowcytometry. Results showed Incubation of leukemic cells (HL-60 and K562 cell lines) with 10 micromol/l AS PS-ODN would significantly reduce the their mRNA levels and in vitro expression of hTERT protein 24 h later, so that the telomerase activity would be significantly down-regulated or inhibited. In conclusion, the hTERT AS PS-ODN is an excellent inhibitor for telomerase activity. PMID- 12095227 TI - Sulphate influx in the erythrocytes of normotensive, diabetic and hypertensive patients. AB - This study aimed to show that modifications in intracellular metabolism are implicated in the pathophysiology of diabetes mellitus and essential hypertension. In fact, total magnesium, calcium, sodium and potassium concentrations, measured in the erythrocytes of normotensive, diabetic and hypertensive patients, have given the following results: a lower intracellular potassium concentration in the erythrocytes of diabetic and hypertensive patients than the erythrocytes of normotensive patients and a more elevated sodium, magnesium, calcium concentrations in the erythrocytes of diabetic and hypertensive patients than the normotensive. Because of the importance of Mg2+ and Ca2+ in metabolic enzyme regulation and their interaction with both Hb and band 3 protein, we examined SO4(2-) kinetic influx in the erythrocytes of normotensive, hypertensive and diabetic patients. The kinetic plots showed different profiles over the three groups and the fluxes were found to be 0.024, 0.061 and 0.072 mmol x (l cells x min)(-1) in normotensive, hypertensive and diabetic patients, respectively. We also found that the Vmax and Km of sulphate influx, obtained by Hofstee plots, increased in the erythrocytes of hypertensive and diabetic patients compared with control cells. In contrast, sulphate influx in the erythrocytes of diabetic and hypertensive patients in the presence of Nifedipine, a calcium antagonist, showed no difference either in the rate constants or in the kinetic profiles, compared to the normotensive control subjects. PMID- 12095229 TI - Discordance between the effect of modulators of calcium on staurosporine-induced apoptosis and staurosporine-induced actin degradation. AB - Staurosporine produced DNA fragmentation characteristic of apoptosis and a dramatic alteration of cell structure that include alterations of cytoskeletal actin and cytoplasmic condensation with vacuolization. These changes were not induced by chelerythrine, a more specific PKC inhibitor than staurosporine. The calcium chelator, BAPTA, significantly reduced staurosporine-induced DNA fragmentation but did not affect staurosporine-induced changes in cytoskeletal actin. These data suggest that DNA fragmentation and actin degradation in apoptosis, induced by staurosporine, are under different regulatory control by calcium. PMID- 12095230 TI - Changes in elastin-binding protein in fibroblasts derived from cardinal ligaments of patients with prolapsus uteri. AB - Prolapsus uteri in pelvic supportive disorders are common in elderly women, and their etiology remains unclear. We examined elastin-binding proteins (EBPs) and binding sites in cultured cardinal ligament fibroblasts derived from elderly patients with prolapsus uteri (HPLiF) and compared them with those from age matched control subjects (HCLiF). Cell attachment to alpha-elastin was significantly lower in HPLiF than in HCLiF. Elastin suppressed the higher proliferative activity at near confluency in HPLiF. The 67-kDa EBP was detectable in HCLiF, whereas HPLiF expressed a 59-kDa EBP. The expression of EBP was significantly lower in HPLiF. The synthetic peptide Val-Gly-Val-Ala-Pro-Gly (VGVAPG), which contains a recognition sequence for the elastin receptor, inhibited the adhesion of HCLiF to alpha-elastin at 10(-5)-10(-4) M, but showed no inhibitory activity on the adhesion of HPLiF at 10(-5) M. These results suggest that fibroblasts derived from elderly women with prolapsus uteri can recognize alpha-elastin through interactions with the low-molecular-size (59-kDa) EBP for the sequence VGVAPG with low affinity and may contribute to the loss of supportive function in uterine connective tissues. PMID- 12095231 TI - Solubility partitioning of C/EBPbeta on the rat hepatocyte nuclear matrix by hydrophobic interactions. AB - The greatest part of nuclear C/EBPbeta (a major 35 kD protein, 30 and 38 kD isoforms) was observed to partition with the nuclear matrix. Cross-linking experiments with formaldehyde suggested that the association reflected the in situ juxtapositioning of C/EBPbeta to nuclear matrix proteins in isolated nuclei. The association of C/EBPbeta with the nuclear matrix resisted RNase and DNase treatment and extraction with protein sulfhydryl reducing agents combined with high ionic strength salt. C/EBPbeta displayed a proclivity to extensively reassemble with the filament-forming nuclear matrix proteins after a cycle of solubilization with urea, followed by its removal by dialysis. These findings suggest that the C/EBPbeta moieties were anchored to the nuclear matrix through hydrophobic protein-protein interactions with the lamins. Subsequent separation of nuclear matrix-associated C/EBPbeta into insoluble, reassembling, and soluble nuclear matrix protein (SNMP) fractions after a cycle of solubilization/reassembly pointed to the sub-partitioning of C/EBPbeta on the nuclear matrix. DNA affinity chromatography using the rat haptoglobin gene cis element and SNMP revealed the binding of p35 during basal transcription, and p35 and p30 during elevated haptoglobin gene transcription in the course of the acute phase (AP) response. It was concluded that the appearance of cis -element-binding p30 in the SNMP fraction resulted from its increased solubility (decreased hydrophobicity) and inability to reassociate with the lamins during urea removal. The observed solubility partitioning of C/EBPbeta on the nuclear matrix framework could represent a level of control of the general availability of regulatory proteins for establishing interactions with DNA. PMID- 12095232 TI - Direct evidence for a role of beta-catenin/LEF-1 signaling pathway in induction of EMT. AB - Epithelial-mesenchymal transformation (EMT) is an important process in development that is characterized by loss of E-cadherin, beta-catenin relocalization, and acquisition of elongated cell shape and ability to invade ECM. beta-catenin has been shown to activate LEF-1 transcription during EMT induced in vitro by c-Fos. Here, we ask whether or not LEF-1 directly introduced into epithelial cells in an adenovirus construct can induce EMT. In normal epithelial cell lines, such as HCE and MDCK cells, that contain functional APC, nuclear beta-catenin induced by exogenous LEF-1 is rapidly exported and EMT is not induced. Leptomycin-B blocks beta-catenin nuclear export, but no EMT occurs due to toxicity. Addition of Wnt-1 to normal epithelial cell lines stabilizes cytoplasmic beta-catenin that LEF-1 then transports to nuclei, causing a small amount of EMT. Our experiments demonstrated, however, that overexpressed LEF-1 upregulates nuclear beta-catenin and promotes dramatic EMT in DLD-1 epithelial tumors that retain nuclear beta-catenin. This EMT is reversible if the LEF-1 virus is removed. Thus, our results demonstrate that LEF-1 can induce EMT directly when its transcription activity is activated by stable nuclear beta catenin. Normal adult epithelial cells appear to use APC to keep beta-catenin out of the nucleus, thereby avoiding pathologies such as metastases due to LEF/beta catenin-induced EMT. PMID- 12095233 TI - The training of cell biologists. PMID- 12095234 TI - Infestation of the blue mussel Mytilus galloprovincialis by the copepod Pseudomyicola spinosus and its relation to size, density, and condition index of the host. AB - The copepod Pseudomyicola spinosus infests marine mussels and other commercial bivalve species. There is a lack of information on the infestation process and on its relationship to size, density, and health of the host. To obtain this information, an infestation study of the copepod in Mytilus galloprovincialis in field conditions was carried out. Results showed that the intensity of infestation is closely related to host size [F((2,810))=198.33; p<0.001], but not to density [F((2,810))=0.96; p<0.38]. After an initial colonization of the host by the infested copepodite stages, an equilibrium between the intensity of the infestation and host size is maintained; mean intensity in small mussels was 1.57, in medium mussels it was 4.23, and in large mussels it was 9.86. Adult stages were observed after a period of 30 days, and ovigerous females were observed after a period of 45 days. Copepods may be found on the branches and mantle (89.5%) and inside the digestive tract (10.5%); in this last case and at the end of the study period some histological disorders were found such as obstruction of the intestine, rupture of the digestive epithelia, and encapsulations around digestive tissues. Most infested mussels showed the lowest meat weight in relation to shell weight; however, the reproductive stage may affect this observation. PMID- 12095235 TI - Bacteria in ovarioles of females from maleless families of ladybird beetles Adalia bipunctata L. (Coleoptera: Coccinellidae) naturally infected with Rickettsia, Wolbachia, and Spiroplasma. AB - Ovarioles were found to be infected with Spiroplasma, Wolbachia, and Rickettsia in Adalia bipunctata females with maleless progeny in different natural populations. Ooplasm was infected with few Wolbachia bacteria. In ooplasm infected by Rickettsia, bacteria were present in small foci. Spiroplasmas were found encapsulated into ooplasm from the wider intercellular spaces between epithelial and oocyte cells. The cytoplasm of follicular epithelia infected with Rickettsia was heavily destroyed, but the nucleus was intact and free from bacteria. The essential feature of follicular epithelium cells from Spiroplasma and Wolbachia infected A. bipunctata females was inclusions of three types: crystalline, filaments, and concentric myelin-like lamellae. Observations of smears prepared from ovaries of A. bipunctata from natural populations revealed a low concentration of bacteria within a microscopy field (less 10 bacteria) in more than 90% of specimens, and only a few ovaries were heavily infected. Two different ways of bacterial invasion of the oocyte are suggested: Spiroplasma like, through the intercellular spaces in the epithelium and Rickettsia-like, through the cytoplasm of follicular epithelium cells. Bacteria were not found in germarium zones and we suggest that each follicle is infected from haemolymph. PMID- 12095237 TI - Effects of gamma irradiation on the development of Trypanosoma rangeli in the vector Rhodnius prolixus. AB - Studies on the effects of gamma radiation on the infectivity of Trypanosoma rangeli (strain H14) for the vector Rhodnius prolixus revealed that (i) the LD(50) (lethal dose for 50% of bugs) for uninfected insects was 4147 rads; (ii) irradiated insects with a dose of 1200 rads subsequently infected with the flagellates exhibited a mortality of 45%, while uninfected irradiated insects showed a mortality of 5%, and infected nonirradiated insects exhibited 10% mortality; (iii) flagellates were present in the hemolymph of irradiated insects 7 days postinfection (p.i.), while in nonirradiated insects the parasites appeared in the hemocoel 18 days p.i.; (iv) T. rangeli infection decreased the number of hemocytes significantly and induced the formation of nodules in the hemolymph of both irradiated and nonirradiated insects; and (v) gamma irradiation affected the ultrastructural organization of the epithelial cells of the small intestine, principally the perimicrovillar membranes and microvilli. In this paper, we discuss the significance of the intestinal microenvironment of R. prolixus with regard to its interaction with T. rangeli. PMID- 12095236 TI - Field and laboratory studies of Ophryoglena sp. (Ciliata: Ophryoglenidae) infection in zebra mussels, Dreissena polymorpha (Bivalvia: Dreissenidae). AB - This study, conducted in the Dnieper-Bug Canal in Belarus, is the first to monitor the seasonal (June-November) dynamics of infection with the parasitic ciliate Ophryoglena sp. in a zebra mussel (Dreissena polymorpha) population. Mean population prevalence and intensity of infection varied, respectively, from 11 to 62% and from 0.9 to 24.1 ciliates/mussel. Mean prevalence was highly correlated with mussel length in mussels <20 mm (R(2)=0.97) and was lower in larger mussels. Mean infection intensity in mussels 1-25 mm long was similarly correlated with their size (R=0.98), reached a maximum in the 20-25 mm size-class, and then sharply decreased, thus providing evidence, albeit limited, that high intensity of infection might be lethal. Transinfection of zebra mussels by Ophryoglena sp. was achieved in the laboratory-a first for a protozoan parasite of D. polymorpha; from an initial complete lack of infection, mean prevalence and intensity rose, respectively, to 86.7% and 8.3 ciliates/mussel. PMID- 12095238 TI - Modulation by eicosanoid biosynthesis inhibitors of immune responses by the insect Manduca sexta to the pathogenic fungus Metarhizium anisopliae. AB - Metarhizium anisopliae conidia (spores) reduced weight gain and caused death when injected into Manduca sexta larvae. When the fungus was co-injected with the eicosanoid biosynthesis inhibitor dexamethasone, larval weight gain was further reduced and mortality increased. These effects were reversed when dexamethasone was given together with the eicosanoid precursor arachidonic acid (AA). Similarly, treatment with other eicosanoid biosynthesis inhibitors (esculetin, phenidone, ibuprofen, and indomethacin) with differing modes of action enhanced the reduction in weight gain caused by mycosis. Injection of M. anisopliae conidia induced nodule formation in vivo; nodule numbers were reduced by dexamethasone, and restored by AA. Incubation of hemocytes with conidia caused microaggregation of hemocytes (indicative of nodule formation) in vitro and this was inhibited by dexamethasone, suggesting that dexamethasone acts directly on hemocytes, although inhibition was only partially reversed by AA. We suggest that the M. sexta immune response to fungal pathogens is normally modulated by physiological systems that include eicosanoid biosynthesis. This is the first demonstration that the virulence of a fungal entomopathogen can be enhanced by compromising the insect host's immune system. PMID- 12095239 TI - Effects of two acanthocephalan parasites on the fecundity and pairing status of female Gammarus pulex (Crustacea: Amphipoda). AB - Acanthocephalan parasites are known to alter the reproductive biology and physiology of their hosts in various ways. In this study we investigated the influence of two acanthocephalan parasites, Pomphorhynchus laevis and Polymorphus minutus, on the fecundity and pairing success of female Gammarus pulex. The results show that P. laevis and P. minutus affect female intermediate host reproduction in different ways. Females infected with P. minutus were totally castrated, whereas those infected with P. laevis only showed reduced fecundity. The oocytes of P. laevis-infected females showed a similar structure to those of uninfected females, although infected females had a higher proportion of oocytes that had failed to reach complete maturity. In comparison, the oocytes of P. minutus-infected females demonstrate a clearly altered structure that suggests a major disruption to the process of vitellogenesis. In the field, males paired more frequently with uninfected females than with infected ones, and is a stronger effect for P. minutus-infected females than P. laevis-infected females. We suggest that the difference in pairing success of P. minutus-infected and P. laevis-infected females is a direct result of the different effects that the two parasites have on female fecundity. PMID- 12095240 TI - Coelomocyte locomotion in the sipunculan Themiste petricola induced by exogenous and endogenous chemoattractants: role of a CD44-like antigen--HA interaction. AB - Cell migration is a key event in the invertebrate immuno-defense system. Microbial products like lipopolysacharide (LPS) and formyl-methyl-leucyl phenylalanine (fMLP) promote cell recruitment to sites of infection. In mammals, complement activation by factors such as zymosan induces C5a production, which influences leukocyte migration. The endogenous factor hyaluronic acid (HA), an extracellular matrix component, also promotes cell migration through its receptor CD44. We evaluated whether coelomocytes from the sipunculan worm T. petricola migrated towards LPS, fMLP, or zymosan treated plasma (ZTP) and if HA was involved in coelomocyte migration and adhesion. We also evaluated if antibodies specific for mouse HA receptor CD44 inhibited any of the effects induced by HA. Using microchemotaxis chambers we found that coelomocytes migrated towards exogenously and endogenously derived chemoattractants. We also observed that HA was a potent chemotactic signal and that coelomocytes adhered strongly to plates coated with LMW-HA but not with HMW-HA. In addition we found that these HA mediated effects were blocked by the monoclonal antibody IM7 directed to mouse CD44, suggesting that a CD44-like cross-reactive antigen might play a role in HA mediated coelomocyte locomotion. PMID- 12095241 TI - Presence of Wolbachia endosymbionts in different silkworm species and races and in their uzi fly parasites. PMID- 12095243 TI - Pathogenic Bacillus from a larva of the Simulium tuberosum species complex (Diptera: Simuliidae). PMID- 12095242 TI - The effect of heavy metal ions and their complexons upon the growth, sporulation and pathogenicity of the entomopathogenic fungus Verticillium lecanii. PMID- 12095244 TI - A venom protein from the endoparasitoid wasp Pimpla hypochondriaca is similar to snake venom reprolysin-type metalloproteases. PMID- 12095245 TI - Alternative method for preservation of mosquito larvae to study binding mechanisms of Bacillus sphaericus binary toxin. PMID- 12095246 TI - Insecticidal activity of the Bacillus sphaericus Mtx1 toxin against Chironomus riparus. PMID- 12095247 TI - Interference between the PHA-4 and PEB-1 transcription factors in formation of the Caenorhabditis elegans pharynx. AB - PHA-4 is a forkhead/winged helix transcription factor that acts as an organ identity factor in the development of the Caenorhabditis elegans pharynx. PEB-1 is a novel DNA-binding protein also involved in pharyngeal morphogenesis. PHA-4 and PEB-1 bind at overlapping sites on the C183 sequence element that controls pharynx-specific expression of the C. elegans myo-2 gene. It has been suggested that PHA-4 and PEB-1 act cooperatively on the C183 sequence. In this study, we test this model and assess the C183-dependent transcriptional activity of PHA-4 and PEB-1, both individually and in combination. We show that PHA-4 and PEB-1 are both modest transcriptional activators in yeast but that co-expression of the two factors does not result in significantly increased expression of a C183-regulated reporter gene. Electrophoretic mobility-shift assays provide no evidence for the formation of a PHA-4/PEB-1 complex in vitro but rather show that PHA-4 and PEB-1 cannot bind C183 simultaneously. As we have reported previously, ectopic expression of PHA-4 in C. elegans causes ectopic expression of a C183-regulated reporter gene. We show that ectopic expression of PEB-1 cannot cause ectopic expression of the same reporter but rather ectopic PEB-1 inhibits reporter gene activation by PHA-4. Overall, our results do not support a model in which PHA-4 and PEB-1 synergize in vivo but rather support a model in which PEB-1 may negatively modulate PHA-4's ability to activate transcription through C183 during formation of the C. elegans pharynx. PMID- 12095248 TI - Structural Modelling of the Sm-like Protein Hfq from Escherichia coli. AB - The Hfq polypeptide of Escherichia coli is a nucleic acid-binding protein involved in the expression of many proteins. Derivation of its three-dimensional structure is important for our understanding of its role in gene regulation at the molecular level. In this study, we combined computational and biophysical analysis to derive a possible structure for Hfq. As a first step towards determining the structure, we searched for possible sequence-structure compatibility, using secondary structure prediction and protein domain and fold recognition methods available on the WEB. One fold, essentially beta sheet in character, the Sm motif of small nuclear ribonucleoproteins, even though it initially fell well below the confidence thresholds, was proposed and further validated by a series of biophysical and biochemical studies. The Hfq hexamer structure was modelled on the human Sm D3B structure using optimised sequence alignments and molecular mechanics methods. This structure accounts for the physico-chemical properties of Hfq and highlights amino acid residues that could interact with RNA. PMID- 12095249 TI - Proteins with class alpha/beta fold have high-level participation in fusion events. AB - Now that complete genome sequences are available for a variety of organisms, the elucidation of potential gene products function is a central goal in the post genome era. Domain fusion analysis has been proposed recently to infer the functional association of the component proteins. Here, we took a new approach to the analysis of the structural features of the proteins involved in fusion events. An exhaustive survey of fusion events within 30 completely sequenced genomes and subsequent structure annotations to the component proteins at a SCOP superfamily level with hidden Markov models was carried out. A domain fusion map was then constructed. The results revealed that proteins with the class alpha/beta fold are frequently involved in fusion events, around 86% of the total 676 assigned single-domain fusion pairs including at least one component protein belonging to the alpha/beta fold class. Moreover, the domain fusion map in our work may offer an attractive framework for designing chimeric enzymes following Nature's lead, and may give useful hints for exploring the evolutionary history of proteins. (c) 2002 Elsevier Science Ltd. PMID- 12095250 TI - A neutral molecule in a cation-binding site: specific binding of a PEG-SH to acetylcholinesterase from Torpedo californica. AB - The crystal structure of acetylcholinesterase from Torpedo californica complexed with the uncharged inhibitor, PEG-SH-350 (containing mainly heptameric polyethylene glycol with a terminal thiol group) is determined at 2.3 A resolution. This is an untypical acetylcholinesterase inhibitor, since it lacks the cationic moiety typical of the substrate (acetylcholine). In the crystal structure, the elongated ligand extends along the whole of the deep and narrow active-site gorge, with the terminal thiol group bound near the bottom, close to the catalytic site. Unexpectedly, the cation-binding site (formed by the faces of aromatic side-chains) is occupied by CH(2) groups of the inhibitor, which are engaged in C-H...pi interactions that structurally mimic the cation-pi interactions made by the choline moiety of acetylcholine. In addition, the PEG-SH molecule makes numerous other weak but specific interactions of the C-H...O and C H...pi types. PMID- 12095251 TI - Widespread and extensive editing of mitochondrial mRNAS in dinoflagellates. AB - We report evidence of extensive substitutional editing of mitochondrial mRNAs in the dinoflagellate species Pfiesteria piscicida, Prorocentrum minimum and Crypthecodinium cohnii, based on a comparison of genomic and corresponding cDNA sequences determined for two mitochondrial DNA-encoded genes, cox1 (cytochrome oxidase subunit 1) and cob (apocytochrome b). In the cox1 mRNA, we identify 72 substitutions at 40 sites in 39 codons, whereas in cob mRNA, we infer 86 editing events at 51 sites in 48 codons. Editing, which takes place in distinct clusters, changes approximately 2% of the total sequence, occurs predominantly at first and second positions of codons, and involves mostly (but not exclusively) A-->G (47%), U-->C (23%) and C-->U (17%) substitutions. In all but four of the 158 cases, editing changes the identity of the specified amino acid. At 21 (cox1) and 26 (cob) sites, the same nucleotide change is observed at the same position in at least two of the species investigated. At about one-third of the sites, editing results in an amino acid change that increases similarity between the dinoflagellate Cox1 and Cob sequences and their homologs in other organisms; presumably editing at these sites is of particular functional significance. Overall, about half of the editing events either maintain or increase similarity between the dinoflagellate protein sequences and their non-dinoflagellate homologs, while a further one-third of the alterations are "dinoflagellate specific" (i.e. they involve a change to an amino acid residue selectively conserved in at least two of the dinoflagellate species at a given position). The nature, pattern and phylogenetic distribution of the inferred edits implies either that more than one type of previously described editing process operates on a given transcript in dinoflagellate mitochondria, or that a mechanistically unique type of mitochondrial mRNA editing has evolved within the dinoflagellate lineage. PMID- 12095252 TI - Kinetic model of DNA replication in eukaryotic organisms. AB - We formulate a kinetic model of DNA replication that quantitatively describes recent results on DNA replication in the in vitro system of Xenopus laevis prior to the mid-blastula transition. The model describes well a large amount of different data within a simple theoretical framework. This allows one, for the first time, to determine the parameters governing the DNA replication program in a eukaryote on a genome-wide basis. In particular, we have determined the frequency of origin activation in time and space during the cell cycle. Although we focus on a specific stage of development, this model can easily be adapted to describe replication in many other organisms, including budding yeast. PMID- 12095253 TI - Evolution of enzymes in metabolism: a network perspective. AB - Several models have been proposed to explain the origin and evolution of enzymes in metabolic pathways. Initially, the retro-evolution model proposed that, as enzymes at the end of pathways depleted their substrates in the primordial soup, there was a pressure for earlier enzymes in pathways to be created, using the later ones as initial template, in order to replenish the pools of depleted metabolites. Later, the recruitment model proposed that initial templates from other pathways could be used as long as those enzymes were similar in chemistry or substrate specificity. These two models have dominated recent studies of enzyme evolution. These studies are constrained by either the small scale of the study or the artificial restrictions imposed by pathway definitions. Here, a network approach is used to study enzyme evolution in fully sequenced genomes, thus removing both constraints. We find that homologous pairs of enzymes are roughly twice as likely to have evolved from enzymes that are less than three steps away from each other in the reaction network than pairs of non-homologous enzymes. These results, together with the conservation of the type of chemical reaction catalyzed by evolutionarily related enzymes, suggest that functional blocks of similar chemistry have evolved within metabolic networks. One possible explanation for these observations is that this local evolution phenomenon is likely to cause less global physiological disruptions in metabolism than evolution of enzymes from other enzymes that are distant from them in the metabolic network. PMID- 12095254 TI - The nuclease A inhibitor represents a new variation of the rare PR-1 fold. AB - Nuclease A (NucA) from Anabaena sp. is a non-specific endonuclease able to degrade single and double-stranded DNA and RNA. The endonucleolytic activity is inhibited by the nuclease A inhibitor (NuiA), which binds to NucA with 1:1 stoichiometry and picomolar affinity. In order to better understand the mechanism of inhibition, the solution structure of NuiA was determined by NMR methods. The fold of NuiA is an alpha-beta-alpha sandwich but standard database searches by DALI and TOP revealed no structural homologies. A visual inspection of alpha-beta alpha folds in the CATH database revealed similarities to the PR-1-like fold (SCOP nomenclature). The similarities include the ordering of secondary structural elements, a single helix on one face of the alpha-beta-alpha sandwich, and three helices on the other face. However, a major difference is in the IV helix, which in the PR-1 fold is short and perpendicular to the I and III helices, but in NuiA is long and parallel to the I and III helices. Additionally, a strand insertion in the beta-sheet makes the NuiA beta-sheet completely antiparallel in organization. The fast time-scale motions of NuiA, characterized by enhanced flexibility of the extended loop between helices III and IV, also show similarities to P14a, which is a PR-1 fold. We propose that the purpose of the PR-1 fold is to form a stable scaffold to present this extended structure for biological interactions with other proteins. This hypothesis is supported by data that show that when NuiA is bound to NucA significant changes in chemical shift occur in the extended loop between helices III and IV. PMID- 12095255 TI - Structural basis for mobility in the 1.1 A crystal structure of the NG domain of Thermus aquaticus Ffh. AB - The NG domain of the prokaryotic signal recognition protein Ffh is a two-domain GTPase that comprises part of the prokaryotic signal recognition particle (SRP) that functions in co-translational targeting of proteins to the membrane. The interface between the N and G domains includes two highly conserved sequence motifs and is adjacent in sequence and structure to one of the conserved GTPase signature motifs. Previous structural studies have shown that the relative orientation of the two domains is dynamic. The N domain of Ffh has been proposed to function in regulating the nucleotide-binding interactions of the G domain. However, biochemical studies suggest a more complex role for the domain in integrating communication between signal sequence recognition and interaction with receptor. Here, we report the structure of the apo NG GTPase of Ffh from Thermus aquaticus refined at 1.10 A resolution. Although the G domain is very well ordered in this structure, the N domain is less well ordered, reflecting the dynamic relationship between the two domains previously inferred. We demonstrate that the anisotropic displacement parameters directly visualize the underlying mobility between the two domains, and present a detailed structural analysis of the packing of the residues, including the critical alpha4 helix, that comprise the interface. Our data allows us to propose a structural explanation for the functional significance of sequence elements conserved at the N/G interface. PMID- 12095256 TI - Solution structure of the pro-hormone convertase 1 pro-domain from Mus musculus. AB - The solution structure of the mouse pro-hormone convertase (PC) 1 pro-domain was determined using heteronuclear NMR spectroscopy and is the first structure to be obtained for any of the domains in the convertase family. The ensemble of NMR derived structures shows a well-ordered core consisting of a four-stranded antiparallel beta-sheet with two alpha-helices packed against one side of this sheet. Sequence homology suggests that the other eukaryotic PC pro-domains will have the same overall fold and most of the residues forming the hydrophobic core of PC1 are highly conserved within the PC family. However, some of the core residues are predicted by homology to be replaced by polar amino acid residues in other PC pro-domains and this may help to explain their marginal stability. Interestingly, the folding topology observed here is also seen for the pro-domain of bacterial subtilisin despite little or no sequence homology. Both the prokaryotic and eukaryotic structures have hydrophobic residues clustered on the solvent-accessible surface of their beta-sheets although the individual residue types differ. In the bacterial case this region is buried at the binding interface with the catalytic domain and, in the eukaryotic PC family, these surface residues are conserved. We therefore propose that the hydrophobic patch in the PC1 pro-domain is involved in the binding interface with its cognate catalytic domain in a similar manner to that seen for the bacterial system. The PC1 pro-domain structure also reveals potential mechanisms for the acid-induced dissociation of the complex between pro- and catalytic domains. PMID- 12095257 TI - Solution structure of the PDZ2 domain from cytosolic human phosphatase hPTP1E complexed with a peptide reveals contribution of the beta2-beta3 loop to PDZ domain-ligand interactions. AB - The solution structure of the second PDZ domain from human phosphatase hPTP1E in complex with a C-terminal peptide from the guanine nucleotide exchange factor RA GEF-2 has been determined using 2D and 3D heteronuclear NMR experiments. Compared to previously solved structures, the hPTP1E complex shows an enlarged interaction surface with the C terminus of the bound peptide. Novel contacts were found between the long structured beta2/beta3 loop of the PDZ domain and the sixth amino acid residue from the C terminus of the peptide. This work underlines the importance of the beta2/beta3 loop for ligand selection by PDZ domains. PMID- 12095258 TI - Native state hydrogen exchange study of suppressor and pathogenic variants of transthyretin. AB - Transthyretin (TTR) is an amyloidogenic protein whose aggregation is responsible for numerous familial amyloid diseases, the exact phenotype being dependent on the sequence deposited. Many familial disease variants display decreased stability in vitro, and early onset pathology in vivo. Only subtle structural differences were observed upon crystallographic comparison of the disease associated variants to the T119M interallelic trans-suppressor. Herein three human TTR single amino acid variant homotetramers including two familial amyloidotic polyneuropathy (FAP) causing variants (V30M and L55P), and a suppressor variant T119M (known to protect V30M carriers from disease by trans suppression) were investigated in a residue-specific fashion by monitoring (2)H (1)H exchange employing NMR spectroscopy. The measured protection factors for slowly exchanging amide hydrogen atoms reveal destabilization of the protein core in the FAP variants, the core consisting of strands A, B, E and G and the loop between strands A and B. The same core exhibits much slower exchange in the suppressor variant. Accelerated exchange rates were observed for residues at the subunit interfaces in L55P, but not in the T119M or V30M TTR. The correlation between destabilization of the TTR core strands and the tendency for amyloid formation supports the view that these strands are involved in amyloidogenicity, consistent with previous (2)H-(1)H exchange analysis of the WT-TTR amyloidogenic intermediate. PMID- 12095259 TI - On the role of periodism in the origin of proteins. AB - Two different views have been proposed for origins of genes (or proteins). One is that primordial genes evolved from random sequences. This view underlies the concept of modern in vitro evolution experiments that functional molecules (even proteins) evolved from random sequence-libraries. On the contrary, the second view reminds that "random sequences" would be an unusual state in which to find RNA or DNA, because it is their inherent nature to yield periodic structures during the course of semi-conservative replication. In this second view, the periodicity of DNA (or RNA) is responsible for emergence of primordial genes. Although recent reports on the variety of periodicities present in proteins, genes and genomes are consistent with the second view, it has yet to be experimentally tested. We assessed the significance of periodicities of DNA in the origin of genes by constructing such periodic DNAs. The results showed that periodic DNA produced ordered proteins at very high rates, which is in contrast to the fact that proteins with random sequences lack secondary structures. We concluded that periodicity played a pivotal role in the origin of many genes. The observation should pave the way for new experimental evolution systems for proteins. PMID- 12095261 TI - Sequence and structure classification of kinases. AB - Kinases are a ubiquitous group of enzymes that catalyze the phosphoryl transfer reaction from a phosphate donor (usually ATP) to a receptor substrate. Although all kinases catalyze essentially the same phosphoryl transfer reaction, they display remarkable diversity in their substrate specificity, structure, and the pathways in which they participate. In order to learn the relationship between structural fold and functional specificities in kinases, we have done a comprehensive survey of all available kinase sequences (>17,000) and classified them into 30 distinct families based on sequence similarities. Of these families, 19, covering nearly 98% of all sequences, fall into seven general structural folds for which three-dimensional structures are known. These fold groups include some of the most widespread protein folds, such as Rossmann fold, ferredoxin fold, ribonuclease H fold, and TIM beta/alpha-barrel. On the basis of this classification system, we examined the shared substrate binding and catalytic mechanisms as well as variations of these mechanisms in the same fold groups. Cases of convergent evolution of identical kinase activities occurring in different folds are discussed. PMID- 12095260 TI - Temperature-dependent dynamics of the villin headpiece helical subdomain, an unusually small thermostable protein. AB - (15)N spin relaxation experiments were used to measure the temperature-dependence of protein backbone conformational fluctuations in the thermostable helical subdomain, HP36, of the F-actin-binding headpiece domain of chicken villin. HP36 is the smallest domain of a naturally occurring protein that folds cooperatively to a compact native state. Spin-lattice, spin-spin, and heteronuclear nuclear Overhauser effect relaxation data for backbone amide (15)N spins were collected at five temperatures in the range of 275-305 K. The data were analyzed using a model-free formalism to determine generalized order parameters, S, that describe the distribution of N-H bond vector orientations in a molecular reference frame. A novel parameter, Lambda=dln(1-S)/dln T is introduced to characterize the temperature-dependence of S. An average value of Lambda=4.5 is obtained for residues in helical conformations in HP36. This value of Lambda is not reproduced by model potential energy functions commonly used to parameterize S. The maximum entropy principle was used to derive a new model potential function that reproduces both S and Lambda. Contributions to the entropy, S(r), and heat capacity, C(r)(p), from reorientational conformational fluctuations were analyzed using this potential energy function. Values of S(r) show a qualitative dependence on S similar to that obtained for the diffusion-in-a-cone model; however, quantitative differences of up to 0.5k, in which k is the Boltzmann constant, are observed. Values of C(r)(p) approach zero for small values of S and approach k for large values of S; the largest values of C(r)(p) are predicted to occur for intermediate values of S. The results suggest that backbone dynamics, as probed by relaxation measurements, make very little contribution to the heat capacity difference between folded and unfolded states for HP36. PMID- 12095262 TI - The structure of Rhodothermus marinus Cel12A, a highly thermostable family 12 endoglucanase, at 1.8 A resolution. AB - Cellulose is one of the most abundant polysaccharides in nature and microorganisms have developed a comprehensive system for enzymatic breakdown of this ubiquitous carbon source, a subject of much interest in the biotechnology industry. Rhodothermus marinus produces a hyperthermostable cellulase, with a temperature optimum of more than 90 degrees C, the structure of which is presented here to 1.8 A resolution. The enzyme has been classified into glycoside hydrolase family 12; this is the first structure of a thermophilic member of this family to have been solved. The beta-jelly roll fold observed has identical topology to those of the two mesophilic members of the family whose structures have been elucidated previously. A Hepes buffer molecule bound in the active site may have triggered a conformational change to an active configuration as the two catalytic residues Glu124 and Glu207, together with dependent residues, are observed in a conformation similar to that seen in the structure of Streptomyces lividans CelB2 complexed with an inhibitor. The structural similarity between this cellulase and the mesophilic enzymes serves to highlight features that may be responsible for its thermostability, chiefly an increase in ion pair number and the considerable stabilisation of a mobile region seen in S. lividans CelB2. Additional aromatic residues in the active site region may also contribute to the difference in thermophilicity. PMID- 12095263 TI - Crystal structure of a transcarbamylase-like protein from the anaerobic bacterium Bacteroides fragilis at 2.0 A resolution. AB - A transcarbamylase-like protein essential for arginine biosynthesis in the anaerobic bacterium Bacteroides fragilis has been purified and crystallized in space group P4(3)2(1)2 (a=b=153.4 A, c=94.8 A). The structure was solved using a single isomorphous replacement with anomalous scattering (SIRAS) and was refined at 2.0 A resolution to an R-factor of 20.6% (R-free=25.2%). The molecular model is trimeric and comprises 960 amino acid residues, two phosphate groups and 422 water molecules. The monomer has the consensus transcarbamylase fold with two structural domains linked by two long interdomain helices: the putative carbamoyl phosphate-binding domain and a binding domain for the second substrate. Each domain has a central parallel beta-sheet surrounded by alpha-helices and loops with alpha/beta topology. The putative carbamoyl phosphate-binding site is similar to those in ornithine transcarbamylases (OTCases) and aspartate transcarbamylases (ATCases); however, the second substrate-binding site is strikingly different. This site has several insertions and deletions, and residues critical to substrate binding and catalysis in other known transcarbamylases are not conserved. The three-dimensional structure and the fact that this protein is essential for arginine biosynthesis suggest strongly that it is a new member of the transcarbamylase family. A similar protein has been found in Xylella fastidiosa, a bacterium that infects grapes, citrus and other plants. PMID- 12095265 TI - DNA methylation analysis in mammalian cells. PMID- 12095266 TI - Identification and resolution of artifacts in bisulfite sequencing. AB - Bisulfite sequencing has become the most widely used application to detect 5 methylcytosine (5-MeC) in DNA, and provides a reliable way of detecting any methylated cytosine at single-molecule resolution in any sequence context. The process of bisulfite treatment exploits the different sensitivity of cytosine and 5-MeC to deamination by bisulfite under acidic conditions, in which cytosine undergoes conversion to uracil while 5-MeC remains unreactive. In this article, we address the more commonly encountered experimental artifacts associated with bisulfite sequencing, and provide methods for the detection and elimination of these artifacts. In particular, we focus on conditions that inhibit complete bisulfite-mediated conversion of cytosines in a target sequence, and demonstrate the necessity of complete protein removal from DNA samples prior to bisulfite treatment. We also include a brief summary of the experimental protocol for bisulfite treatment and tips for designing polymerase chain reaction (PCR) primers to amplify from bisulfite-treated DNA. PMID- 12095267 TI - Methylation sequencing from limiting DNA: embryonic, fixed, and microdissected cells. AB - It is frequently useful to determine the methylation state of samples containing limited amounts of DNA such as from embryos, or from fixed tissue samples in which DNA is degraded or difficult to isolate. By modification of the standard protocols for DNA preparation and bisulfite treatment, it is possible to obtain DNA methylation sequence data for such samples. We present methods for bisulfite treatment of embryos, fixed sections, and samples obtained by laser capture microdissection, and discuss the additional experimental considerations required when working with small numbers of cells or degraded DNA samples. PMID- 12095268 TI - Conversion-specific detection of DNA methylation using real-time polymerase chain reaction (ConLight-MSP) to avoid false positives. AB - Methylated cytosines appear as sequence variations following bisulfite treatment and polymerase chain reaction (PCR) amplification. By using methylation-specific PCR (MSP), it is possible to detect methylated sequences in a background of unmethylated DNA with a high level of sensitivity. MSP is frequently used to identify methylated alleles in carcinogenesis, and may be combined with the TaqMan real-time PCR system, which uses fluorescence-based detection of amplification products during the amplification phase of the PCR and increases the sensitivity of detection (MethyLight). Sequences that have been incompletely converted during the bisulfite treatment are frequently coamplified during MSP, resulting in an overestimation of DNA methylation. The presence of amplified sequences originating from partially unconverted material may be determined by sequencing or by restriction digests or Southern blots of MSPs. Alternately, we have developed a method where the PCR and conversion assay are combined within a single TaqMan reaction by using an additional fluorescent probe directed against unconverted DNA (ConLight-MSP). We recommend that MSP detection always should include a step to detect unconverted DNA to avoid overestimation of the frequency or level of methylated DNA in the sample. PMID- 12095269 TI - Profiling DNA methylation by melting analysis. AB - The idea of modifying DNA with bisulfite has paved the way for a variety of polymerase chain reaction (PCR) methods for accurately mapping 5-methylcytosine at specific genes. Bisulfite selectively deaminates cytosine to uracil under conditions where 5-methylcytosine remains unreacted. Following conventional PCR amplification of bisulfite-treated DNA, original cytosines appear as thymine while 5-methylcytosines appear as cytosine. Because the relative thermostability of a DNA duplex increases with increasing content of G:C base pairs, PCR products originating from DNA templates with different contents of 5-methylcytosine differ in melting temperature, i.e., the temperature required to convert the double helix into random coils. We describe two methods that resolve differentially methylated DNA sequences on the basis of differences in melting temperature. The first method integrates PCR amplification of bisulfite-treated DNA and subsequent melting analysis by using a thermal cycler coupled with a fluorometer. By including in the reaction a PCR-compatible, fluorescent dye that specifically binds to double-stranded DNA, the melting properties of the PCR product can be examined directly in the PCR tube by continuous fluorescence monitoring during a temperature transition. The second method relies on resolution of alleles with different 5-methylcytosine contents by analysis of PCR products in a polyacrylamide gel containing a gradient of chemical denaturants. Optimal resolution of differences in melting temperature is achieved by a special design of PCR primers. Both methods allow resolution of "heterogeneous" methylation, i.e., the situation where the content and distribution of 5-methylcytosine in a target gene differ between different molecules in the same sample. PMID- 12095270 TI - Quantitative methylation analysis using methylation-sensitive single-nucleotide primer extension (Ms-SNuPE). AB - Methylation-sensitive single-nucleotide primer extension (Ms-SNuPE) is a technique that allows for rapid and simultaneous quantitation of the degree of methylation at several CpG sites. Treatment of genomic DNA with sodium bisulfite is used to convert unmethylated cytosine to uracil while leaving 5-methylcytosine unchanged. A strand-specific polymerase chain reaction product is then generated to provide a suitable DNA template for quantitative methylation analysis using Ms SNuPE. Single-nucleotide primer extension is performed with oligonucleotide(s) designed to hybridize immediately upstream of the CpG site(s) being analyzed. The Ms-SNuPE technique can be adapted for high-throughput methylation analysis and therefore represents a novel approach for rapid quantitation of cytosine methylation suitable for a wide range of biological investigations. PMID- 12095271 TI - Screening for and analysis of methylation differences using methylation-sensitive single-strand conformation analysis. AB - Methylation-sensitive single-strand conformation analysis (MS-SSCA) is a method of screening for methylation changes at CpG sites in a region of DNA. After bisulfite modification, the region of interest is amplified using primers specific for bisulfite-modified sequences. The amplified products are denatured and run on a nondenaturing polyacrylamide gel. The sequence differences caused by methylation lead to the formation of different secondary structures (conformers) with different mobilities. MS-SSCA is a convenient and rapid method for screening large numbers of samples for methylation. Individual bands can readily be isolated and sequenced allowing more detailed analysis of methylation changes. In this article, we present a protocol for MS-SSCA and outline strategies for the design of primers for amplifying bisulfite-modified DNA sequences. PMID- 12095272 TI - Analysis of the methylation status of imprinted genes based on methylation specific polymerase chain reaction combined with denaturing high-performance liquid chromatography. AB - A procedure for the analysis of the methylation status of imprinted genes is described. The method offers a rapid and reliable alternative to conventional methods such as Southern blots and methylation-specific polymerase chain reaction (PCR) (i.e., allele-specific methylation-specific PCR). The efficient resolution of the differentially methylated alleles is demonstrated for three human imprinted genes: SNRPN, LIT1 (alias KCNQ1OT1), and H19. Abnormal imprinting of SNRPN is associated with the Angelman/Prader-Willi syndromes, and that of LIT1 and H19 with the Beckwith-Wiedemann syndrome. The method is based on methylation specific PCR followed by denaturing high-performance liquid chromatography (MSP/DHPLC). Briefly, genomic DNA is initially subjected to an in vitro bisulfite treatment, whereby unmethylated cytosines are deaminated. Subsequent PCR amplifications, using primers specific for modified DNA, are aimed at DNA segments that show parent-of-origin-specific methylation. PCR conditions are chosen that allow an efficient amplification of both alleles. The PCR products representing the two alleles are identical in size; they differ, however, at a number of positions within the amplified DNA segment. The DHPLC analysis allows very efficient resolution of the two populations of PCR products, providing qualitative and quantitative results. PMID- 12095273 TI - Restriction landmark genome scanning. AB - Restriction landmark genome scanning (RLGS) is a quantitative approach that is uniquely suited for simultaneously assessing the methylation status of thousands of CpG islands. RLGS separates radiolabeled NotI fragments (or other CpG containing restriction enzyme fragments) in two dimensions and allows distinction of single-copy CpG islands from multicopy CpG-rich sequences. The methylation sensitivity of the endonuclease activity of NotI provides the basis for differential methylation analysis, and NotI sites occur primarily in CpG islands and genes. RLGS has been used to identify novel imprinted genes, novel targets of DNA amplification and methylation in human cancer, and to identify deletion, methylation, and gene amplification in a mouse model of tumorigenesis. Such massively parallel analyses are critical for pattern recognition within and between tumor types, and for estimating the overall influence of CpG island methylation on the cancer cell genome. RLGS is also a useful method for integrating methylation analyses with high-resolution gene copy number analyses. PMID- 12095274 TI - Identification of DNA methylation differences during tumorigenesis by methylation sensitive arbitrarily primed polymerase chain reaction. AB - The ability to detect methylation changes associated with oncogenic transformation is of critical importance in understanding how DNA methylation may contribute to tumorigenesis. We have developed a simple and reproducible fingerprinting method called methylation-sensitive arbitrarily primed polymerase chain reaction (AP-PCR) to screen for DNA methylation changes. This technique relies on digesting genomic DNA with methylation-sensitive and -insensitive restriction enzymes (e.g., HpaII and MspI) prior to AP-PCR amplification. Matched normal and tumor DNAs were compared to identify differential methylation. After the PCR products were resolved on high-resolution polyacrylamide gels, regions of genomic DNA that showed hypo- and hypermethylation associated with tumors were detected. These fragments were then isolated, cloned, and sequenced. Novel CpG islands were found to be frequently hypermethylated in bladder and colon tumors. We have demonstrated that this technique is a rapid and efficient method that can be used to screen for altered methylation patterns in genomic DNA and to isolate specific sequences associated with these changes. PMID- 12095275 TI - Measurement of genome wide DNA methylation by reversed-phase high-performance liquid chromatography. AB - A method to determine the extent of cytosine methylation in DNA is described. The technique involves the enzymatic hydrolysis of DNA to its deoxyribonucleotide components and subsequent separation and quantification of the nucleotides by isocratic reversed-phase high-performance liquid chromatography (HPLC). The system gives highly reproducible results and, under suitable conditions, is capable of measuring 5-methylcytosine levels in as little as 1 microg of DNA. PMID- 12095276 TI - Use of CpG island microarrays to identify colorectal tumors with a high degree of concurrent methylation. AB - We provide a comprehensive description of our microarray-based technique for the simultaneous detection of multiple CpG islands in cancer. Amplicons from tumor and control samples were pools of differentially methylated CpG island fragments hybridized to a panel of approximately 8000 CpG island tags. Data analysis identified 694 CpG island loci hypermethylated in a group of 14 colorectal tumors. The Stanford hierarchical cluster algorithm segregated the tumors into two subgroups, one of which exhibited a high level of concurrent hypermethylation while the other had little or no methylation. This is in agreement with previous observations of a CpG island methylation phenotype present in colorectal tumors. The present study demonstrates that this microarray-based technique is useful in classifying tumors according to their methylation profiles. PMID- 12095277 TI - Analysis of complex methylation data. AB - The development of multiple DNA methylation analysis techniques, including higher throughput assays, has resulted in data structures of increasing complexity and diversity. Here, we discuss the general principles of DNA methylation analysis and propose a nomenclature for the various types of methylation analysis. We briefly outline several DNA methylation analysis techniques and discuss how these different technologies affect the structure of the resulting methylation data. We then describe the basic statistics and bioinformatic principles relevant to the analysis of simple and complex methylation data. PMID- 12095278 TI - DNA methylation and epigenetic inheritance. AB - Mammalian cell lines silence genes at low frequency by the methylation of promoter sequences. These silent genes can be reactivated at high frequency by the demethylating agent 5-azacytidine (5-aza-CR). The inactive and active epigenetic states of such genes are stably inherited. A method for silencing genes is now available. It involves treatment of permeabilized cells with 5 methyl deoxycytidine triphosphate (5-methyl dCTP) which is incorporated into DNA. The methylation of promoter sequences has been confirmed using the bisulfite genomic sequencing procedure. Methylated oligonucleotides homologous to promoter sequences might be used to specifically target and silence given genes, but results so far have not been conclusive. Treatments that silence or reactivate genes by changing DNA methylation can be referred to as epimutagens, as distinct from mutagens that act by changing DNA sequences. The epimutagen 5-aza-CR reactivates genes but has little mutagenic activity, whereas standard mutagens (such as ethyl methane sulfonate and ultraviolet light) have little reactivation activity. Nevertheless, much more information is required about the effects of DNA-damaging agents in changing DNA methylation and gene activity and also about the role of epimutations in tumor progression. PMID- 12095279 TI - Utilization of antisense oligonucleotides to study the role of 5-cytosine DNA methyltransferase in cellular transformation and oncogenesis. AB - A large body of data point toward 5-cytosine DNA methyltransferase 1 (DNMT1) as a critical component of oncogenic programs. The study of the role of DNMT1 in cancer has been hindered by the lack of specific inhibitors. A different approach to study the role of DNMT1 in cancer is to use sequence-specific antisense oligonucleotides against DNMT1 mRNA. This paper discusses methods used to identify sequence-specific antisense oligonucleotides and to assess their DNA methylation inhibitory properties. Antisense oligonucleotides are applied to determine whether DNMT1 plays a causal role in specific cancer models ex vivo as well as in vivo. PMID- 12095295 TI - Hypothesis-driven science. PMID- 12095296 TI - Gene therapy and the germ line. PMID- 12095297 TI - Baculovirus as mammalian cell expression vector for gene therapy: an emerging strategy. AB - The monopoly of insect cells to host baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) as a eukaryotic gene expression system has been shattered with the growing evidence that it also infects mammalian cells in culture. Although AcMNPV fails to replicate in vertebrate cells, it does express alien genes with levels of expression that are dependent on the strength of the promoter used to drive transcription of the foreign gene. It also has been reported that the recombinant AcMNPV enters human hepatic cells in culture preferentially and specifically in comparison with the other mammalian cells of different origin and sources. This has resulted in the use of AcMNPV as a potent mammalian cell delivery system as a xenovector for gene therapy, more precisely liver-specific gene delivery in vitro and in vivo. PMID- 12095298 TI - Neurotrophin-3 transduction attenuates cisplatin spiral ganglion neuron ototoxicity in the cochlea. AB - Ototoxicity is a major dose-limiting side effect of cisplatin chemotherapy for cancer patients. We previously demonstrated in vitro that herpes simplex type 1 (HSV-1) amplicon-mediated delivery of a neurotrophin-3 (NT-3)/myc chimera protects spiral ganglion neurons (SGNs) in murine cochlear cultures from cisplatin-induced ototoxicity. To extend these findings, a newly constructed amplicon vector (HSVnt-3myc/SV40lac) that expresses the NT-3myc chimera and the Escherichia coli beta-galactosidase (lacZ) reporter gene under separate transcriptional control was initially tested in vitro and then was delivered to the cochlea of aged mice that were subsequently treated with cisplatin. Successful transduction with the new amplicon was observed in vitro as determined by its capacity to infect SGNs and to express NT-3myc mRNA and protein. To determine whether amplicon-directed NT-3myc overexpression could abrogate the ototoxicity in vivo, two groups of aged mice (CBA) were inoculated with HSVnt 3myc/SV40lac or control vector, HSVSV40lac, preceding administration of cisplatin. Cochleas inoculated with HSVnt-3myc/SV40lac harbored significantly greater numbers of surviving SGNs and showed lower incidence of cisplatin-induced apoptosis or necrosis than those injected with the control virus. These data demonstrate that HSV amplicon-mediated NT-3 delivery can attenuate the ototoxic actions of cisplatin in the peripheral auditory system of the aged mouse. The potency of NT-3 in SGN neuroprotection suggests that in vivo neurotrophin-based gene therapy is a promising preventative treatment for chemical-induced hearing disorders, and potentially for hearing degeneration due to normal aging. PMID- 12095299 TI - Biodistribution and toxicity studies of VSVG-pseudotyped lentiviral vector after intravenous administration in mice with the observation of in vivo transduction of bone marrow. AB - Lentiviral vectors can confer high levels of gene transfer and transgene expression in a variety of cell types. However, the biodistribution and toxicity after intravenous administration have not been reported. To address these issues of biodistribution and toxicity, an HIV-1-based vector, HR'cmvGFP, was administered to normal BALB/c mice by tail-vein injection. Nine different organs and bone marrow were evaluated by real-time quantitative PCR (QPCR) assay capable of a broad range of quantitation (5-log fold) to detect as few as one copy of the green fluorescent protein gene (GFP) per 10(5) cells. Four days after vector administration, high levels of transgene and gene expression were observed in liver, spleen, and bone marrow in all animals. By 40 days after injection, GFP levels had decreased in liver and spleen, but bone marrow exhibited a consistently high level of transgene. This finding was consistent with the increase in both GFP frequency and expression levels observed in peripheral blood by fluorescence-activated cell-sorting (FACS) analysis. Between 0 and 1% transgene was detected in all other organs. No significant pathologic lesions were found attributable to vector in any of the tissues examined. The observation of bone marrow transduction after intravenous vector administration suggests the possibility of an in vivo approach to stem cell gene therapy. PMID- 12095300 TI - Optimizing aptamer activity for gene therapy applications using expression cassette SELEX. AB - RNA aptamers against a variety of clinically relevant target proteins have been generated. For example, we previously isolated an RNA aptamer that inhibits the function of the E2F family of transcription factors that play a critical role in the control of cell proliferation. However, the development of this and other aptamers for gene therapy applications has been complicated by the fact that expression of RNA aptamers in the context of flanking sequences can inhibit the ability of an aptamer to fold into its functional conformation. Insertion of the E2F aptamer into a tRNA expression cassette resulted in the production of high levels of chimeric tRNA that contains a misfolded and inactive aptamer in transfected mammalian cells. To overcome this problem, we randomized the sequence flanking the aptamer and selected for chimeric tRNAs that retained high affinity binding to E2F1. This expression cassette SELEX strategy yielded RNAs that bind E2F with high affinity (IC50 of 15 nM) and which can be expressed at high levels in mammalian cells. Moreover, these chimeric tRNA-E2F aptamers are functional and can inhibit E2F-mediated transactivation by up to 80% in human 293 cells. Expression cassette SELEX should greatly facilitate the use of aptamers for a variety of gene therapy applications. PMID- 12095301 TI - Deoxyribonuclease I-like III is an inducible macrophage barrier to liposomal transfection. AB - Extra- and intracellular nucleases are predicted to decrease the in vivo efficiency of liposomal transfection. DNASE1 (D1) has been proposed as the main nuclease barrier, yet liposome-complexed DNA and in vitro lipofection are generally immune to D1. In contrast, medium conditioned by the macrophage enzyme DNASE1-like 3 (DNASE1L3 or D3) erects a potent in vitro barrier to liposomal transfection. Although homologous to D1 over its amino-terminal sequence, D3 has a distinct, highly basic carboxy terminus, which resembles polylysine stretches often found in polycationic liposomal reagents. If this domain is truncated from D3, the resulting enzyme has more nuclease activity against naked DNA ("free DNA" nuclease activity), yet does not block transfection. C-terminal fusion of this domain to D1 forms a chimeric protein able to block transfection. D3 can be immunodetected in both serum and macrophage lysates. Macrophage-conditioned medium contains both "free DNA"-nuclease activity and the ability to block transfection, and by zymogram only a 28-kDa DNASE, consistent by size with D3, is present. Thus, medium containing D3 confers to cells an in vivo shield to the nuclear acquisition of exogenous DNA. Modulation and further elucidation of this activity are likely to have importance for both gene therapy and autoimmune disorders. PMID- 12095302 TI - Comparison of surfactant and perfluorochemical liquid enhanced adenovirus mediated gene transfer in normal rat lung. AB - Both surfactant- and perfluorochemical (PFC)-based vehicles enhance adenovirus mediated gene transfer in the lung. To compare the relative effects of surfactant and PFC liquid, we infected orotracheally intubated Sprague-Dawley rats with 4 x 10(9) pfu of an E1a(-)/E3(-) adenovirus expressing either an Escherichia coli lacZ (AdlacZ) mini-gene or no cDNA (Adnull). Surfactant-mediated delivery was achieved via instillation of four, 200-microl aliquots of virus suspended in a 50% surfactant (Survanta) vehicle over a 15-minute period. PFC rats received virus in 100 microl of saline followed by instillation of the PFC liquid FC-75 (10 cc/kg body weight) over a 2- to 3- minute period. Lungs were collected 3 days later for measurement of beta-galactosidase (beta-gal) expression and indices of inflammation. Both PFC liquid and surfactant-based vehicles produced widespread beta-gal expression and increased total beta-gal activity over that observed with instillation of vector alone. Both vehicles comparably increased bronchoalveolar lavage fluid (BALF), total cell counts, neutrophils, total protein, and IFN(gamma). FC-75 was also associated with increased BALF IL1beta. In conclusion, surfactant and FC-75 are similarly effective vehicles for adenovirus-mediated gene transfer to the lung. PMID- 12095303 TI - Tumor growth inhibition by interferon-alpha using PEGylated protein or adenovirus gene transfer with constitutive or regulated expression. AB - Inducible synthesis and secretion of therapeutic proteins following gene transfer could be a viable strategy to deliver biopharmaceuticals that currently require parenteral administration. Evaluating the protein pharmacokinetics and biological responses generated by different delivery modalities will provide a better understanding of the advantages and disadvantages of each strategy. The interferon-alpha (IFN-alpha) family of proteins, used clinically for infectious and malignant diseases, has a short half-life, and IFN-alpha therapy requires frequent administration of the drug by injection. Subcutaneous xenograft tumors were inhibited by weekly administration of polyethylene glycol modified (PEGylated) IFN-alpha protein or by a single administration of an adenovirus constitutively expressing IFN-alpha (IACB). Both treatment modalities inhibited tumor growth in a dose-dependent manner, suggesting that increasing exposure to IFN-alpha could result in effective tumor control. A single adenovirus that encodes the components necessary for tetracycline induction (IADR) expressed IFN alpha in a ligand-dependent manner. Adding doxycycline to the drinking water of mice treated intravenously with the inducible adenovirus IADR inhibited tumor growth by 85% compared with mice that were not given doxycycline. The correlation between serum IFN-alpha concentration and the degree of tumor growth inhibition did not depend on the delivery technology used. It is likely that it will be feasible to control expression of IFN-alpha by oral administration of small molecule drugs after gene delivery to induce therapeutic concentrations of proteins. PMID- 12095304 TI - On the kinetics of polyplex endocytic trafficking: implications for gene delivery vector design. AB - Synthetic gene therapy vectors must be designed to safely and efficiently escort DNA from outside the cell to the nucleus and to overcome several physical barriers that are obstacles to internalization, escape from endocytic vesicles, movement through the cytoplasm, and transport into the nucleus. By providing a firm foundation for polymer design, a quantitative understanding of polymer mediated gene delivery mechanisms may allow more efficient and timely design of new vectors. We have used a flow-cytometry-based assay for investigating endocytic trafficking by quantitation of polyplex pH microenvironments. We investigated polyethylenimine (PEI)- and poly-L-lysine (PLL)-DNA trafficking, with and without the endosomotropic agent chloroquine. PLL-DNA complexes appear to be localized to early endocytic vesicles and are not trafficked to lysosomes. Further, chloroquine appears to facilitate PLL-mediated gene delivery by a mechanism other than buffering of endocytic compartments. Additionally, PEI does not appear to buffer endocytic compartments, but requires exposure to an acidic environment for efficient gene delivery. PMID- 12095305 TI - Antisense gene therapy of brain cancer with an artificial virus gene delivery system. AB - Therapeutic genes are delivered to the nuclear compartment of cancer cells following intravenous administration with a non-immunogenic "artificial virus" gene delivery system that uses receptor-specific monoclonal antibodies (MAb) to navigate the biological barriers between the blood and the nucleus of the cancer cell. Mice implanted with intracranial U87 human glial brain tumors are treated with a nonviral expression plasmid encoding antisense mRNA against the human epidermal growth factor receptor gene (EGFR). The plasmid DNA is packaged within the interior of polyethylene glycol-modified (PEGylated) immunoliposomes, and delivered to the brain tumor with MAbs that target the mouse transferrin receptor (TRFR) and the human insulin receptor (INSR). The mouse TRFR MAb enables transport across the tumor vasculature, which is of mouse brain origin, and the INSR MAb causes transport across the plasma membrane and the nuclear membrane of the human brain cancer cell. The lifespan of the mice treated weekly with an intravenous administration of the EGFR antisense gene therapy packaged within the artificial virus is increased 100% relative to mice treated either with a luciferase gene or with saline. PMID- 12095306 TI - Noninvasive, repetitive, quantitative measurement of gene expression from a bicistronic message by positron emission tomography, following gene transfer with adenovirus. AB - Gene therapy protocols are hampered by the inability to monitor the location, magnitude, and duration of ectopic gene expression following DNA delivery. Consequently, it is difficult to establish quantitative correlations and/or causal relationships between therapeutic gene expression and phenotypic responses in treated individuals. One approach to monitor "therapeutic gene" expression indirectly is to incorporate reporter genes that can be imaged in vivo into bicistronic transcription units, along with the therapeutic genes. Expression of the dopamine D2 receptor (D2R) and herpes simplex virus thymidine kinase (HSV1 TK) can both be monitored, in vivo, by positron-emission tomography (PET). We created ad.DTm, an adenovirus containing a cytomegalovirus (CMV) early promoter driven transcription unit, in which the D2R gene is placed proximal to an encephalomyocarditis virus internal ribosomal entry site (IRES) and a modified HSV1-tk gene is placed distal to the IRES. Following intravenous ad.DTm injection into mice, correlated hepatic D2R and HSV1-sr39tk PET reporter gene expression was demonstrated. Repeated microPET scanning quantitated both D2R-dependent sequestration of a positron-emitting ligand and HSV1-TK-dependent sequestration of a positron-emitting product. It is possible, in living mice, to investigate noninvasively and to measure quantitatively and repeatedly correlated expression of two coding regions from a bicistronic transcription unit over a 3-month period following adenovirus delivery. PMID- 12095307 TI - Kinetics of fluorescence expression in nonhuman primates transplanted with GFP retrovirus-modified CD34 cells. AB - Downregulation and loss of proviral expression have been demonstrated to occur in a variety of in vitro studies and in mouse models. Here we evaluated the kinetics of proviral expression after transplantation in a competitive repopulating model in the baboon. Transgene persistence and green fluorescent protein (GFP) expression in peripheral blood leukocytes (PBL) were analyzed in four animals by semiquantitative PCR and flow cytometry for up to 80 weeks (range 17-80). All animals were transplanted with cells transduced with EGFP or EYFP reporters driven by Moloney murine leukemia virus (MoMuLV) or a modified promoter/enhancer, (MND) respectively. Simultaneous dual-color analysis of fluorescence levels in granulocyte and lymphocyte subsets following hematopoietic reconstitution demonstrated progressive loss of fluorescence intensity occurring predominantly early after transplant in cells transduced with both retrovirus backbones and at serial time points. In addition, we carried out PCR analysis of DNA extracted from sorted EGFP(-)/EYFP(-) cells and confirmed the presence of cells genetically marked by either vector in this population, indicating the persistence of cells that have downregulated or lost retroviral gene expression. In comparison to mouse studies, however, we did not detect substantial differences between MND and MoMuLV backbones. PMID- 12095308 TI - An adenoviral expression system for AAV rep78 using homologous recombination. AB - The construction and amplification of adenoviral (Ad) vectors expressing biologically active transgenes that are cytotoxic or inhibit Ad replication can be extremely difficult, if not impossible. In this study, we harnessed the ability of Ad genomes to undergo efficient homologous recombination to reconstitute the adeno-associated virus (AAV) rep78 gene, a cytotoxic gene that strongly inhibits Ad replication, which was divided between two parental, first generation Ad vectors. A functional open reading frame was generated by recombination only upon co-infection of both parental vectors and after the onset of viral DNA replication. We were able to amplify both parental rep78 vectors to normal titers without any signs of inhibition or toxicity and could use them to generate progeny vectors containing a functional rep78 gene without any Ad genes. Using this vector recombination system in AAV rescue assays demonstrated that no Ad protein was essential for Rep78 mediated rescue of AAV ITR flanked DNA from plasmid or Ad backbones; the amount of rescue product generated was substantially greater in the presence of Ad infection; neither cellular nor viral DNA replication was necessary for rescue to occur; and progeny vector genomes were efficiently co-replicated along with conventional, first-generation Ad vectors. PMID- 12095309 TI - Restoration of a functional open reading frame by homologous recombination between two adenoviral vectors. AB - In this study, we examined the ability of adenoviral (Ad) vectors to undergo homologous recombination. The lacZ gene was divided between two parental, first generation vectors such that neither encoded a functional product but both shared 494 bp in common. The open reading frame could only be restored by homologous recombination. We observed beta-galactosidase activity only upon co-infection of both parental vectors and after the onset of viral DNA replication, creating a delay in expression of 24-36 hours in HeLa cells. At peak efficiency, this recombination vector system resulted in beta-galactosidase activity levels 100x above background and just 18x less than a conventional, first-generation vector in HeLa cells. After recombination, the resultant progeny vector genomes containing reconstituted expression cassettes were devoid of all viral genes and contained two packaging signals. These progeny genomes were efficiently packaged, could be separated from their parental vectors based on their lighter buoyant densities in CsCl gradients, and were subsequently used as functional gene transfer vectors. This novel recombination vector system should be useful for transferring large transgenes (because the carrying capacity of two Ad vectors can be exploited) or expressing any cytotoxic or Ad replication inhibitory protein (because the parental vectors exhibit no background expression). PMID- 12095310 TI - Improved method of recombinant AAV2 delivery for systemic targeted gene therapy. AB - A major hurdle in most current gene therapy modalities is the ability to transduce target tissues at very high efficiencies that ultimately lead to therapeutic levels of transgene expression. We have developed a novel method of recombinant adeno-associated virus 2 (rAAV) delivery that results in increased vector transduction efficiencies using microspheres reversibly conjugated to rAAV vectors. We hypothesize that conjugation to microspheres should result in a higher effective concentration of vector as well as longer relative exposure time of vector to target cells as it moves through the tissue vasculature. In vitro experiments demonstrate that the same level of transduction seen with free vector can be achieved using 1% of vector when conjugated to microspheres. In addition, using magnetic microspheres, the region of infection can be targeted. In vivo, we demonstrate that microsphere-mediated delivery of rAAV vector results in higher transduction efficiencies than delivery with free vector alone when administered either intramuscularly or intravenously. Furthermore, we demonstrate targeting of transgene expression to specific tissues by retention of microsphere-bound vector in the capillary bed. These studies demonstrate a novel method to deliver rAAV vectors more effectively that could prove to be a successful alternative mode of virus-mediated human gene therapy. PMID- 12095311 TI - Blockerette-ligated capture T7-amplified RT-PCR, a new method for determining flanking sequences. AB - We have developed a highly sensitive PCR-based technique termed blockerette ligated capture T7-amplified reverse-transcription PCR (BCT-RT-PCR), which can be used to characterize unknown proviral flanking sequences from a broad range of samples and depends only on knowing the retroviral sequence. This method incorporates several essential elements to make it both sensitive and specific, including a "blockerette" linker, magnetic capture of target sequences, and exponential replication of potentially rare sequences using a nested promoter for T7 RNA polymerase, followed by nested RT-PCR. This linkage of methods was designed to increase sensitivity by decreasing DNA complexity in favor of specific amplification. The resulting PCR products can be directly sequenced to determine integration sequences. We have successfully determined the integration sequences from as little as 30 pg of provirus-containing DNA in the background of 30 ng of untransduced DNA, representing a 0.1% transduction rate. We also show this technique to have single-cell resolution even in the background of 5000 cells. We describe here for the first time the combined use of BCT-RT-PCR and laser-capture microscopy (LCM) for precise isolation of retrovirally transduced cells followed by determination of the 3' retroviral flanking sequence at the single-cell level. This method will aid significantly in determining clonality both in transplant experiments and in the generation of clonal cell populations. PMID- 12095312 TI - Evidence of CFTR function in cystic fibrosis after systemic administration of 4 phenylbutyrate. AB - Most individuals with cystic fibrosis (CF) carry one or two mutations that result in a maturation defect of the full-length protein. One such mutation, deltaF508, results in a mutant membrane glycoprotein that fails to progress to the apical membrane, where the wild-type protein normally functions as a cyclic AMP regulated chloride channel. 4-Phenylbutyrate (Buphenyl), an orally bioavailable short chain fatty acid, modulates heat shock protein expression and restores maturation of the deltaF508 protein in vitro and in vivo. We performed a randomized, double-blind, placebo-controlled, dose-escalation and safety study of Buphenyl in 19 adults with CF (homozygous deltaF508) to test the hypothesis that Buphenyl would be safe, well-tolerated, and associated with an increase in chloride transport in nasal epithelia. Three dose levels (20, 30, or 40 g divided t.i.d.) of drug or placebo were given for 1 week. Serial measurements of chloride transport by nasal potential difference (NPD) testing and metabolic safety testing were performed. A maximum tolerated dose of 20 g was defined based on minimal adverse reactions, the safety profile, and a statistically significant induction of chloride transport that was maximal by day 3. This short-term phase I/II study demonstrates proof of principle that modulation of deltaF508 CFTR biosynthesis and trafficking is a viable therapeutic approach for cystic fibrosis. PMID- 12095313 TI - Increased vascularity detected by digital subtraction angiography after VEGF gene transfer to human lower limb artery: a randomized, placebo-controlled, double blinded phase II study. AB - Vascular endothelial growth factor (VEGF) gene therapy may be useful for the treatment of lower-limb ischemia. The objectives of this study were to evaluate safety and angiographic and hemodynamic responses of local catheter-mediated VEGF gene therapy in ischemic lower-limb arteries after percutaneous transluminal angioplasty (PTA). For this study, we recruited patients with chronic lower-limb ischemia and atherosclerotic infrainguinal occlusion or stenosis suitable for PTA. In the study, 18 patients received 2x10(10) plaque-forming units (pfu) VEGF adenovirus (VEGF-Ad), 17 patients received VEGF-plasmid/liposome (VEGF-P/L; 2000 microg of VEGF plasmid, 2000 microl of DOTMA:DOPE), and 19 control patients received Ringer's lactate at the angioplasty site. Digital subtraction angiography (DSA) was used to evaluate vascularity before, immediately after, and 3 months after the PTA. Clinical follow-up data, basic laboratory tests, and ankle-brachial index (ABI) were evaluated. Primary endpoint was DSA analysis of vascularity, and secondary endpoints were restenosis rate, Rutherford class, and ABI after 3 months follow-up. No major gene transfer-related side effects or differences in laboratory tests were detected between the study groups. However, anti-adenovirus antibodies increased in 61% of the patients treated with VEGF-Ad. For the primary endpoint, follow-up DSA revealed increased vascularity in the VEGF-treated groups distally to the gene transfer site (VEGF-Ad P=0.03, VEGFP/L P=0.02) and in the VEGF-Ad group in the region of the clinically most severe ischemia (P=0.01). As for the secondary endpoints, mean Rutherford class and ABI showed statistically significant improvements in the VEGF-Ad and VEGF-P/L groups, but similar improvements were also seen in the control patients. We conclude that catheter-mediated VEGF gene therapy is safe and well tolerated. Angiography demonstrated that VEGF gene transfer increased vascularity after PTA in both VEGF Ad- and VEGF-P/L-treated groups. PMID- 12095314 TI - Properly designed modular asymmetric synthesis for enantiopure sulfinamide auxiliaries from N-sulfonyl-1,2,3-oxathiazolidine-2-oxide agents. AB - Simple and practical asymmetric synthesis of functionally differentiated aminoindanol based endo-N-sulfonyl 1,2,3-oxathiazolidine-2-oxide as sulfinyl transfer agents are developed. The importance of these new and unique sulfinyl transfer reagents are exemplified by the expedient production of several sulfinamide ligands, including either enantiomer of (R)-tert-butanesulfinamide in excellent yields and enantiopurities. PMID- 12095315 TI - Regio- and enantiospecific rhodium-catalyzed allylic etherification reactions using copper(I) alkoxides: influence of the copper halide salt on selectivity. AB - The transition metal-catalyzed allylic etherification represents a fundamentally important cross-coupling reaction for the construction of allylic ethers. We have developed a new regio- and enantiospecific rhodium-catalyzed allylic etherification of acyclic unsymmetrical allylic alcohol derivatives using copper(I) alkoxides derived from primary, secondary and tertiary alcohols. This study demonstrates that the choice of copper(I) halide salt is crucial for obtaining excellent regio- and enantiospecificity, providing another example of the effect of halide ions in asymmetric transition metal-catalyzed reactions. Finally, the ability to alter the reactivity of the alkali metal alkoxides in this manner may provide a useful method for related metal-catalyzed cross coupling reactions involving heteroatoms. PMID- 12095317 TI - Highly stereoselective intramolecular hydroamination/cyclization of conjugated aminodienes catalyzed by organolanthanides. AB - Efficient intramolecular hydroamination/cyclization of primary and secondary conjugated aminodienes can be effected by using organolanthanide precatalysts of the type Cp'2LnCH(TMS)2 (Cp' = eta5-Me5C5; Ln = La, Sm, Y; TMS = SiMe3) and CGCSmN(TMS)2 (CGC = Me2Si(eta5-Me4C5)(tBuN)). The transformation proceeds cleanly (>/= 90% conversion) at 25-60 degrees C with good rates and high regioselectivities, and with electronic effects leading to significant rate enhancements. Some features of the reaction parallel monosubstituted aminoalkene hydroamination/cyclization, including rate law (zero order in [aminodiene]), and rate enhancements observed with larger lanthanide ionic radii and/or more open catalyst ligation structures. Good to excellent diastereoselectivity is obtained in the synthesis of 2,5-trans-disubstituted pyrrolidines (80% de) and 2,6-cis disubstituted piperidines (99% de) with using the corresponding alpha-methyl aminodiene precursors. Formation of 2-(prop-1-enyl)piperidine with the chiral C1 symmetric precatalyst (S)-Me2Si(OHF)(CpR*)SmN(TMS)2 (OHF = eta5 octahydrofluorenyl; Cp = eta5-C5H3; R* = (-)-menthyl) proceeds with up to 69% ee. PMID- 12095316 TI - A phototriggered molecular spring for aerobic catalytic oxidation reactions. AB - The first catalytic oxidation reactions with cofacial bisporphyrins using molecular oxygen as the terminal oxidant are presented. The photocatalytic reactions proceed under mild conditions (ambient temperature and pressure) without the need for a co-reductant. A systematic reactivity study of the photooxidation of dimethyl sulfide (DMS) with a homologous series of bisiron(III) mu-oxo porphyrins containing dibenzofuran, xanthene, or no bridge reveals that the facility of these processes is markedly dependent on the vertical flexibility of the catalyst to provide a phototriggered molecular spring. PMID- 12095318 TI - Fluorine- and trimethylsilyl-containing phenoxy--imine Ti complex for highly syndiotactic living polypropylenes with extremely high melting temperatures. AB - A fluorine- and trimethylsilyl-containing phenoxy-imine titanium complex was synthesized and the structure was determined by an X-ray analysis. The complex on activation with MAO initiates highly controlled syndiospecific living propylene polymerization to form extremely high Tm syndiotactic polypropylenes (Mw/Mn = 1.05-1.08, Tm = 156-152 degrees C) at 0 or 25 degrees C. Moreover, at 50 degrees C, the complex afforded monodisperse syndiotactic polypropylene with very high Tm's of 149, 150 degrees C. In contrast, complexes having a t-Bu group instead of the silyl group provided lower tacticity polymers with much lower Tm's. In addition, we revealed the substituent effect that plays a key role for the highly controlled syndiospecific polymerization displayed by the catalyst. PMID- 12095319 TI - Tandem intramolecular silylformylation-crotylsilylation: highly efficient synthesis of polyketide fragments. AB - Polyketide fragments may be rapidly and efficiently assembled in the tandem intramolecular silylformylation-crotylsilylation of alkenes and alkynes. The reactions establish up to three new stereocenters with good-to-excellent diastereoselectivity, and the use of the reaction in an iterative sense is demonstrated. In addition, a new dihydrosilane alcoholysis reaction has been developed, leading to a highly efficient sequence. PMID- 12095320 TI - Layered paving of vesicular nanoparticles formed with cerasome as a bioinspired organic-inorganic hybrid. AB - A three-dimensional packed vesicular assembly was successfully prepared by using an organic-inorganic hybrid, the Cerasome. This assembly was achieved by using an alternate layer-by-layer adsorption technique with the anionic and the cationic Cerasome derived from corresponding organoalkoxysilane amphiphiles. Adsorption quantities of each Cerasome layer were evaluated by employing a quartz crystal microbalance. The surface structure of the Cerasome paving on a substrate obtained in this way was observed by atomic force microscopy. The Cerasome particles closely packed like a stone pavement were clearly observed in both layers. In addition, the difference in the particle size for each layer indicates the cationic and anionic Cerasomes undoubtedly formed the layer-by-layer assembly. The layered paving of the vesicular nanoparticles was seen in every layer at least up to ten adsorption steps. PMID- 12095321 TI - The enantioselective organocatalytic 1,4-addition of electron-rich benzenes to alpha,beta-unsaturated aldehydes. AB - The first enantioselective organocatalytic alkylation of electron-rich benzene rings with alpha,beta-unsaturated aldehydes has been accomplished. The use of iminium catalysis has provided a new strategy for the enantioselective construction of benzylic stereogenicity, an important chiral synthon for natural product and medicinal agent synthesis. The (2S,5S)-5-benzyl-2-tert butylimidazolidinone amine catalyst has been found to mediate the conjugate addition of a wide variety of substituted and unsubstituted anilines to unsaturated aldehydes. A diverse spectrum of aldehyde substrates can also be accommodated in this new organocatalytic transformation. While catalyst quantities of 10 mol % were generally employed in this study, successful alkylations conducted with catalyst loadings as low as 1 mol % are described. PMID- 12095322 TI - The iconoclastic dynamics of the 1,2,6-heptatriene rearrangement. AB - CASSCF(8,8)/6-31G* and AM1-SRP direct dynamics trajectory calculations have been run on the rearrangement of 1,2,6-heptatriene to 3-methylene-1,5-hexadiene. They show that the experimental results of Roth et al. on this reaction can be explained without the need to invoke a concerted, pericyclic mechanism. Instead, bifurcation occurs at the transition state for conversion of the reactant to 2 methylenecyclohexane-1,4-diyl. Some trajectories leaving the transition state do enter the PES local minimum for the intermediate, but others, differing only in the phases of the real-frequency vibrational modes, bypass the intermediate and proceed over a second transition state to the product. A significant fraction of the trajectories that do enter the biradical local minimum proceed on to the product in <500 fs, despite the fact that the minimum is some 12 kcal/mol deep at the CASSCF level. This nonstatistical behavior seems to be due in part to a resonance between key C-H bending and C-C stretching vibrations in the intermediate. PMID- 12095323 TI - Bistable ferroelectric liquid crystal photoswitch triggered by a dithienylethene dopant. AB - The spontaneous polarization (PS) of a ferroelectric liquid crystal is modulated reversibly by photocyclization of the dopant 1,2-bis[5'-(4' '-heptyloxyphenyl)-2' methylthien-3'-yl]perfluorocyclopentene. The magnitude of PS photomodulation increases with dopant concentration up to 3 mol %, and the resulting photoswitch is fatigue resistant and bistable. To the best of our knowledge, this is the first example of a bistable ferroelectric liquid crystal photoswitch to be reported in the literature. PMID- 12095324 TI - Ruthenium-catalyzed cycloaddition of propargylic alcohols with phenol derivatives via allenylidene intermediates: catalytic use of the allenylidene ligand as the C(3) unit. AB - Novel ruthenium-catalyzed cycloaddition of propargylic alcohols with 2-naphthols and phenols bearing electron-donating groups via allenylidene intermediates has been developed to give the corresponding 1H-naphtho[2,1-b]pyrans and 4H-1 benzopyrans, respectively, in moderate to excellent yields with complete regioselectivity. PMID- 12095325 TI - Molecular recognition in methanol: the first example of hydrogen-bond-mediated self-association of a calix[4]arene in polar, protic solvent. AB - Tetraalaninecalixarene was prepared by coupling of tetraaminocalix[4]arene with alanine. It dimerizes in methanol, providing the first example of a substituted calixarene that undergoes self-association through hydrogen bonding in polar, protic solvent. The association constant in 24:1 MeOH:H2O was determined to be 29 000 M-1. Addition of arginine or lysine results in disruption of the dimer and formation of a 1:1 complex between the amino acid and the tetraalaninecalixarene. The preparation of a peptidocalixarene that associates in polar solvent opens new doors for the use of calixarenes for molecular recognition in biologically relevant environments. PMID- 12095326 TI - A short synthetic route to (+)-austamide, (+)-deoxyisoaustamide, and (+) hydratoaustamide from a common precursor by a novel palladium-mediated indole --> dihydroindoloazocine cyclization. AB - The first synthesis of (+)-austamide (1), (+)-deoxyisoaustamide (2), and (+) hydratoaustamide (10) by a very direct route is described (Scheme 1). Starting from tryptophan methyl ester (3) intermediate 5 is generated in two steps in >98% overall yield. The key step in the synthesis is a novel cyclization of 5 involving organopalladium intermediates which gives the dihydroazocine 6. From this key intermediate the target structures are accessible in just a few steps as shown in Scheme 1. The remarkable conversion of 5 --> 6 can be rationalized by the mechanistic pathway shown in Scheme 2 that involves a multistep sequence which includes palladation, cyclization, and rearrangement. PMID- 12095327 TI - A 20-A-thick interwoven sheet consisting of nanotubes. AB - Slow diffusion of AgNO3 with Me2Si(4-Py)2 yields an interwoven 2-nm-thick sheet consisting of the building block [Ag3(Me2Si(4-Py)2)4](NO3)3.H2O. The interweaving induces unique nanotunnels with a 16 x 18 A2 cross section with a 7 x 8 A2 square pore. The anion NO3- of the skeletal sheet can be reversibly exchanged with appropriately sized anions without the collapse of its structural integrity. The compound has a melting point (140 degrees C) and exists as the first 2D ionic liquid up to 193 degrees C. PMID- 12095328 TI - Probing polymer/polymer interfaces. AB - Infrared-visible sum frequency generation spectroscopy (SFG) has been used to study the interface between poly(vinyl-N-octadecylcarbamate-co-vinyl acetate) (Comb) and deuterated or hydrogenated polystyrene (dPS or hPS) films. Strong methyl symmetric and Fermi resonance bands associated with the alkyl side chains of the Comb polymer are observed in the SFG spectra. In addition, for Comb/hPS spectra, symmetric and asymmetric vibration modes of phenyl groups are observed. The presence of asymmetric modes indicates the phenyl rings are tilted with respect to the interface normal. PMID- 12095329 TI - Gas-phase metal trications in protic solvent complexes. AB - In the first observation of triply charged metal ions microsolvated by protic ligands, gas-phase complexes of 11 different metal trications with diacetone alcohol were generated using electrospray ionization. These species were found to exist for at least five to eight ligands, depending on the metal. For smaller sizes, charge reduction to dications occurs spontaneously via dissociative proton transfer. This is the largest minimum size reported for any multiply charged metal cation/ligand pair. PMID- 12095330 TI - A strategy for improving the room-temperature luminescence properties of Ru(II) complexes with tridentate ligands. AB - Several ruthenium(II) complexes with new tridentate polypyridine ligands have been prepared, and their photophysical properties have been studied. The new tridentate ligands are tpy-modified systems (tpy = 2,2':6',2' '-terpyridine) in which aromatic substituents designed to be coplanar with the tpy moiety are introduced, with the aim of enhancing delocalization in the acceptor ligand of the potentially luminescent metal-to-ligand charge-transfer (MLCT) state and increasing the MLCT-MC energy gap (MC = metal-centered excited state). Indeed, the Ru(II) complexes obtained with this new family of tridentate ligands exhibit long-lived luminescence at room temperature (up to 200 ns). The enhanced luminescence properties of these complexes support this design strategy and are superior to those of the model Ru(tpy)22+ compound and compare favorably with those of the best Ru(II) complexes with tridentate ligands reported so far. PMID- 12095331 TI - Self-assembly of cobalt nanoparticle rings. AB - Weakly ferromagnetic cobalt nanoparticles can assemble spontaneously into nanosized "bracelets" when dispersed in organic solvents containing resorcinarenes as surfactants. Bracelet self-assembly occurs in solution and is directed by magnetic dipolar interactions, whereas nanoparticle rings with larger diameters are produced by evaporation-driven flow on wetted surfaces. PMID- 12095332 TI - Optically detected magnetic resonance in the photoexcited triplet states of Ti(IV) and Zr(IV) cylopentadienyl complexes. AB - The class of compounds (RCp)2MX2, where M is a group IV metal, Cp is cyclopentadienyl, R is an alkyl, and X is a halide, has been of continuing interest as precursors for olefin coordination polymerization catalysts. In this communication, we demonstrate that the technique of optically detected magnetic resonance (ODMR) reveals subtle differences in the composition of the frontier molecular orbitals associated with the nature of the alkyl substituents on the Cp rings. PMID- 12095333 TI - The vibrational reorganization energy in pentacene: molecular influences on charge transport. AB - The reorganization energy in pentacene is reported on the basis of a joint experimental and theoretical study of pentacene ionization using high-resolution gas-phase photoelectron spectroscopy, semiempirical intermediate neglect of differential overlap calculations, and first-principles correlated quantum mechanical calculations at MP2 and density functional theory levels. The reorganization energy upon positive ionization of pentacene is determined both experimentally and theoretically to be remarkably low. This is one key element that allows one to rationalize the extremely high hole mobilities recently measured in pentacene single crystals. PMID- 12095334 TI - Strained silacycles in organic synthesis: a new reagent for the enantioselective allylation of aldehydes. AB - A new reagent for the enantioselective allylation of aliphatic aldehydes has been developed. The reagent is easily prepared in a single step from commercially available materials and may be stored without significant decomposition. The reactivity of the reagent is attributed to strain-release Lewis acidity. PMID- 12095335 TI - A chemoselective reduction of alkynes to (E)-alkenes. AB - The trans reduction of all types of alkynes to give (E)-olefins is achieved through a two-stage trans hydrosilylation and protodesilylation. Reaction of an alkyne and a silane with the ruthenium catalyst [Cp*Ru(MeCN)3]PF6 results in clean hydrosilylation to give only the (Z)-trans addition product at ambient temperature with catalyst loadings of 1-5 mol %. The crude vinylsilane products are then protodesilylated by the action of cuprous iodide and TBAF at rt-35 degrees C. The reaction is compatible with many sensitive functional groups and provides a general trans-alkyne reduction not possible by other means. PMID- 12095336 TI - Structure-specific DNA cleavage on surfaces. AB - The structure-specific invasive cleavage reaction is a useful means for sensitive and specific detection of single nucleotide polymorphisms, or SNPs, directly from genomic DNA without a need for prior target amplification. A new approach integrating this invasive cleavage assay and surface DNA array technology has been developed for potentially large-scale SNP scoring in a parallel format. Two surface invasive cleavage reaction strategies were designed and implemented for a model SNP system in codon 158 of the human ApoE gene. The upstream oligonucleotide, which is required for the invasive cleavage reaction, is either co-immobilized on the surface along with the probe oligonucleotide or alternatively added in solution. The ability of this approach to unambiguously discriminate a single base difference was demonstrated using PCR-amplified human genomic DNA. A theoretical model relating the surface fluorescence intensity to the progress of the invasive cleavage reaction was developed and agreed well with experimental results. PMID- 12095338 TI - Electron-induced switching of the supramolecular chirality of optically active polythiophene aggregates. AB - A chiral regioregular polythiophene (PT), poly[3-[4-((R)-4-ethyl-2-oxazolin-2 yl)phenyl]thiophene] (poly-1), forms chiral aggregates which exhibit a unique induced circular dichroism (ICD) in the pi-pi transition region derived from the supramolecular chirality in the presence of various poor solvents or metal salts in chloroform. We report here that the chirality of such supramolecular aggregates can be switched ("on" and "off") through electron transfer. We have found that upon the addition of copper(II) trifluoromethanesulfonate [Cu(OTf)(2)] to the chiral aggregates of poly-1 in a chloroform-acetonitrile mixture, the ICD disappears because of the oxidative doping of the poly-1 main chain, while a further addition of amines such as triethylenetetramine (TETA) induces undoping of the poly-1 which results in the reappearance of the ICDs. Therefore, the supramolecular chirality of the poly-1 assemblies was reversibly controlled by the addition or removal of an electron from the poly-1 main chain. This may be the first example of a reversible supramolecular chirality switch on chiral PT aggregates. We investigated the mechanism of the chirality switch through the doping and undoping process on the polymer main chain by means of absorption and CD spectroscopies, ESR, cyclic voltammetry, X-ray diffraction, and AFM measurements. PMID- 12095337 TI - Slow exchange in the chromophore of a green fluorescent protein variant. AB - Green fluorescent protein and its mutants have become valuable tools in molecular biology. They also provide systems rich in photophysical and photochemical phenomena of which an understanding is important for the development of new and optimized variants of GFP. Surprisingly, not a single NMR study has been reported on GFPs until now, possibly because of their high tendency to aggregate. Here, we report the (19)F nuclear magnetic resonance (NMR) studies on mutants of the green fluorescent protein (GFP) and cyan fluorescent protein (CFP) labeled with fluorinated tryptophans that enabled the detection of slow molecular motions in these proteins. The concerted use of dynamic NMR and (19)F relaxation measurements, supported by temperature, concentration- and folding-dependent experiments provides direct evidence for the existence of a slow exchange process between two different conformational states of CFP. (19)F NMR relaxation and line shape analysis indicate that the time scale of exchange between these states is in the range of 1.2-1.4 ms. Thermodynamic analysis revealed a difference in enthalpy (Delta)H(0) = (18.2 +/- 3.8) kJ/mol and entropy T(Delta)S(0) = (19.6 +/- 1.2) kJ/mol at T = 303 K for the two states involved in the exchange process, indicating an entropy-enthalpy compensation. The free energy of activation was estimated to be approximately 60 kJ/mol. Exchange between two conformations, either of the chromophore itself or more likely of the closely related histidine 148, is suggested to be the structural process underlying the conformational mobility of GFPs. The possibility to generate a series of single-atom exchanges ("atomic mutations") like H --> F in this study offers a useful approach for characterizing and quantifying dynamic processes in proteins by NMR. PMID- 12095339 TI - Metal ion-catalyzed nucleic acid alkylation and fragmentation. AB - Nucleic acid microarrays are a growing technology in which high densities of known sequences are attached to a substrate in known locations (addressed). Hybridization of complementary sequences leads to a detectable signal such as an electrical impulse or fluorescence. This combination of sequence addressing, hybridization, and detection increases the efficiency of a variety of genomic disciplines including those that profile genetic expression, search for single nucleotide polymorphisms (SNPs), or diagnose infectious diseases by sequencing portions of microbial or viral genomes. Incorporation of reporter molecules into nucleic acids is essential for the sensitive detection of minute amounts of nucleic acids on most types of microarrays. Furthermore, polynucleic acid size reduction increases hybridization because of increased diffusion rates and decreased competing secondary structure of the target nucleic acids. Typically, these reactions would be performed as two separate processes. An improvement to past techniques, termed labeling-during-cleavage (LDC), is presented in which DNA or RNA is alkylated with fluorescent tags and fragmented in the same reaction mixture. In model studies with 26 nucleotide-long RNA and DNA oligomers using ultraviolet/visible and fluorescence spectroscopies as well as high-pressure liquid chromatography and mass spectrometry, addition of both alkylating agents (5-(bromomethyl)fluorescein, 5- or 6-iodoacetamidofluorescein) and select metal ions (of 21 tested) to nucleic acids in aqueous solutions was critical for significant increases in both labeling and fragmentation, with >or=100-fold increases in alkylation possible relative to metal ion-free reactions. Lanthanide series metal ions, Pb(2+), and Zn(2+) were the most reactive ions in terms of catalyzing alkylation and fragmentation. While oligonucleotides were particularly susceptible to fragmentation at sites containing phosphorothioate moieties, labeling and cleavage reactions occurred even without incorporation of phosphorothioate moieties into the RNA and DNA target molecules. In fact, LDC conditions were found in which RNA could be fragmented into its component monomers, allowing simultaneous sequencing from both the 5'- and the 3'-termini by mass spectrometry. The results can be explained by alkylation of the (thio)phosphodiester linkages to form less hydrolytically stable (thio)phosphotriesters, which then decompose into 2',3'-cyclic phosphate (or 2' phosphate) and 5'-hydroxyl terminal products. Analysis of fragmentation and alkylation products of Mycobacterium tuberculosis (Mtb) ribosomal RNA (rRNA) transcripts by polyacrylamide gel electrophoresis was consistent with the model studies. Building upon these results, I found that products from Mtb rRNA amplification products were processed with fluorescent reporters and metal ions in a single reaction milieu for analysis on an Affymetrix GeneChip. Mild conditions were discovered which balanced the need for aggressive alkylation and the need for controlled fragmentation, advantageously yielding GeneChip results with greater than 98% of the nucleotides reported correctly relative to reference sequences, results sufficient for accurately identifying Mtb from other Mycobacterium species. Thus, LDC is a new, straightforward, and rapid aqueous chemistry that is based on metal ion-catalyzed alkylation and alkylation catalyzed fragmentation of nucleic acids for analysis on microarrays or other hybridization assays and that, possibly, has utility in similar processing of other appropriately functionalized biomolecules. PMID- 12095340 TI - Hydrogenation of carbon dioxide catalyzed by ruthenium trimethylphosphine complexes: the accelerating effect of certain alcohols and amines. AB - A trace amount of alcohol cocatalyst and a stoichiometric amount of base are required during the hydrogenation of CO(2) to formic acid catalyzed by ruthenium trimethylphosphine complexes. Variation of the choice of alcohol and base causes wide variation in the rate of reaction. Acidic, nonbulky alcohols and triflic acid increase the rate of hydrogenation an order of magnitude above that which can be obtained with traditionally used methanol or water. Similarly, use of DBU rather than NEt(3) increases the rate of reaction by an order of magnitude. Turnover frequencies up to 95,000 h(-1) have now been obtained, and even higher rates should be possible using the cocatalyst and amine combinations identified herein. Preliminary in situ NMR spectroscopic observations are described, and the possible roles of the alcohol and base are discussed. PMID- 12095341 TI - Reactivity and selectivity of the N-acetyl-Glu-P-1, N-acetyl-Glu-P-2, N-acetyl MeIQx, and N-acetyl-IQx nitrenium ions: comparison to carbocyclic N-arylnitrenium ions. AB - The model ultimate carcinogens 1a-d, related to the metabolites of the food derived carcinogenic heterocyclic amines Glu-P-1, Glu-P-2, MeIQx, and IQx, spontaneously decompose in neutral aqueous solution to generate the heterocyclic nitrenium ions, 2a-d. The less reactive esters 1a and 1b also undergo acid catalyzed ester hydrolysis to generate the corresponding hydroxamic acids at pH <2, while the more reactive 2c and 2d are prone to rearrangement in nonaqueous solvents. The reactions of the nitrenium ions with AcO(-), HPO(4)(2-), N(3)(-), and 2'-deoxyguanosine (d-G) were characterized in aqueous solution by using a combination of competitive trapping methods and product isolation and identification. The reactions with N(3)(-) and d-G generally follow patterns previously established for carbocyclic nitrenium ions, but the reactions with AcO(-) and HPO(4)(2-) are unusual. Similar reactions have previously only been reported for heterocyclic 1-alkyl-2-imidazolium ions. The N(3)(-)/solvent selectivities of these ions (5.1 x 10(6) M(-1) for 2a, 2.3 x 10(6) M(-1) for 2b, 1.2 x 10(5) M(-1) for 2c, and 5.2 x 10(4) M(-1) for 2d) are comparable to those of highly selective carbocyclic nitrenium ions. If k(az) for these ions is diffusion limited at ca. 5 x 10(9) M(-1) s(-1) the aqueous solution lifetimes of these ions range from 10 micros (2d) to 1 ms (2a). These ions are also highly selective for trapping by d-G, but comparisons to other nitrenium ions show that they are 10- to 50-fold less selective for trapping by d-G than they would be if both the N(3)(-) and d-G reactions were diffusion limited. This is not a consequence of their heterocyclic structures. Several carbocyclic ions show similar behavior. The relatively inefficient trapping of 2c and 2d by d-G may account for the observation of the unusual minor N-2 d-G adduct that is isolated for both of these nitrenium ions, but has not previously been observed for the reactions of other nitrenium ions with monomeric d-G. PMID- 12095342 TI - Highly enantioselective intra- and intermolecular [2 + 2] photocycloaddition reactions of 2-quinolones mediated by a chiral lactam host: host-guest interactions, product configuration, and the origin of the stereoselectivity in solution. AB - The [2 + 2] photocycloaddition of 4-alkoxy-2-quinolones was conducted in the presence of the chiral lactams 5 or ent-5. At -60 degrees C in toluene as the solvent the intramolecular reaction of quinolones 6 and 8 as well as the intermolecular photocycloaddition of various alkenes 13 to quinolone 12 proceeded with excellent enantioselectivity (81-98% ee) and in high yields (61-89%). Styrene (13d) reacted sluggishly in the intermolecular reaction (29% yield, 83% ee). The absolute configuration of the intramolecular photocycloaddition products 7 and 9 was elucidated by single-crystal X-ray crystallography of the corresponding diastereomeric N-menthyloxycarbonyl derivatives. The relative configuration of the intermolecular photocycloaddition products 14 and 15 was assigned on the basis of NOESY experiments and on crystallographic evidence. The differentiation of the enantiotopic faces in the prochiral quinolones 6, 8, and 12 can be explained by assuming a coordination of these substrates to the lactams 5 or ent-5 via two hydrogen bonds. Upon binding to 5 the si-face is shielded by the bulky tetrahydronaphthalene backbone, and the re-face is exposed to an intra- or intermolecular attack. On the basis of the association constant (K(a)) for the coordination of quinolone to host 5 an interpretation of the observed enantiomeric excess has been put forward. The parent quinolone 17 was employed as substrate for microcalorimetric and NMR titration experiments. From the data obtained for K(a) and DeltaH(a) the expected enantiomeric excess was calculated for two given temperatures (-15 and -60 degrees C). The calculated values fit the observed data within reasonable limits and prove that two-point hydrogen bonding can be sufficient to achieve a preparatively useful face differentiation in solution phase photochemistry. PMID- 12095343 TI - Stereochemical effects in supramolecular self-assembly at surfaces: 1-D versus 2 D enantiomorphic ordering for PVBA and PEBA on Ag(111). AB - We present investigations on noncovalent bonding and supramolecular self-assembly of two related molecular building blocks at a noble metal surface: 4-[trans-2 (pyrid-4-yl-vinyl)]benzoic acid (PVBA) and 4-[(pyrid-4-yl-ethynyl)]benzoic acid (PEBA). These rigid, rodlike molecules comprising the same complementary moieties for hydrogen bond formation are comparable in shape and size. For PVBA, the ethenylene moiety accounts for two-dimensional (2-D) chirality upon confinement to a surface; PEBA is linear and thus 2-D achiral. Molecular films were deposited on a Ag(111) surface by organic molecular beam epitaxy and characterized by scanning tunneling microscopy. At low temperatures (around 150 K), both species form irregular networks of flat lying molecules linked via their endgroups in a diffusion-limited aggregation process. In the absence of kinetic limitations (adsorption or annealing at room temperature), hydrogen-bonded supramolecular assemblies form which are markedly different. With PVBA, enantiomorphic twin chains in two mirror-symmetric species running along a high-symmetry direction of the substrate lattice form by diastereoselective self-assembly of one enantiomer. The chirality signature is strictly correlated between neighboring twin chains. Enantiopure one-dimensional (1-D) supramolecular nanogratings with tunable periodicity evolve at intermediate coverages, reflecting chiral resolution in micrometer domains. In contrast, PEBA assembles in 2-D hydrogen-bonded islands, which are enantiomorphic because of the orientation of the supramolecular arrangements along low-symmetry directions of the substrate. Thus, for PVBA, chiral molecules form 1-D enantiomorphic supramolecular structures because of mesoscopic resolution of a 2-D chiral species, whereas with PEBA, the packing of an achiral species causes 2-D enantiomorphic arrangements. Model simulations of supramolecular ordering provide a deeper understanding of the stability of these systems. PMID- 12095345 TI - Adjacent versus opposite type di-aromatic ring-fused phthalocyanine derivatives: synthesis, spectroscopy, electrochemistry, and molecular orbital calculations. AB - A series of adjacent and opposite type di-aromatic ring-fused phthalocyanines (Pc's) of varying size have been prepared and characterized spectroscopically and electrochemically, and most of their properties have been reasonably reproduced by molecular orbital (MO) calculations. The adjacent isomers alone were obtained preferentially by using a diphthalonitrile unit linked via a short aryl chain. The main results are summarized as follows. (i) The Q-band shifts to longer wavelength and its intensity increases, but the degree of change decreases, with increasing molecular size. On the bases of the experiments and MO calculations, setting the size of the effect of benzene directly fused to the tetraazaporphyrin (TAP) skeleton at unity, the effect of the second and third benzene units is roughly about 0.75-0.80 and 0.48 +/- 0.06, respectively. As a result of this, among compounds having an isomeric pi-system, the Q-band of a D(4h) type species lies at longer wavelength than those of adjacently and oppositely di-aromatic ring-fused species. (ii) The Q-band of adjacently substituted species does not split appreciably, while that of the oppositely substituted species splits substantially, the extent having a parallel relationship with the ratio of long to short axes in the molecule. In general, the larger the ratio, the larger the splitting. (iii) The Q-band of oppositely dibenzo-fused and bis(dialkyl) substituted TAP does not show explicit splitting because of the large coefficients of the carbons substituted with alkyl groups in the MOs. (iv) Interestingly, the first oxidation in adjacently and oppositely dibenzo-fused CoTAP occurs at the cobalt and ligand, respectively, although they are isomers to each other. PMID- 12095344 TI - Palladium-catalyzed cross-coupling reaction of bis(pinacolato)diboron with 1 alkenyl halides or triflates: convenient synthesis of unsymmetrical 1,3-dienes via the borylation-coupling sequence. AB - The synthesis of 1-alkenylboronic acid pinacol esters via a palladium-catalyzed cross-coupling reaction of bis(pinacolato)diboron (pin(2)B(2), pin = Me(4)C(2)O(2)) with 1-alkenyl halides or triflates was carried out in toluene at 50 degrees C in the presence of KOPh (1.5 equiv) and PdCl(2)(PPh(3))(2)-2Ph(3)P (3 mol %). The borylation of acyclic and cyclic 1-alkenyl bromides and triflates was achieved in high yields with complete retention of configuration of the double bonds. The method was applied to the one-pot synthesis of unsymmetrical 1,3-dienes via the borylation-coupling sequence. PMID- 12095346 TI - Mono-aromatic ring-fused versus adjacently di-aromatic ring-fused tetraazaporphyrins: regioselective synthesis and their spectroscopic and electrochemical properties. AB - A method which preferentially produces adjacently di-aromatic ring-substituted tetraazaporphyrins (TAPs) has been developed, and their electrochemical and spectroscopic properties have been studied and compared with those of the corresponding series of mono-aromatic ring-fused TAPs. Mono-aromatic ring-fused TAPs show a split Q-band, and the splitting energy increases with increasing size of the aromatic ring. In addition, for the split Q-bands, the relative intensity of the band at longer wavelength decreases with increasing molecular size of the fused aromatics, compared with the shorter wavelength band. In the di-aromatic ring-fused TAPs, this kind of splitting is not seen, and only a shift of the band is observed. The intensity and band position of the split or unsplit Q-bands are quantitatively evaluated by simultaneous band deconvolution analysis, using both electronic absorption and magnetic circular dichroism spectra. The preparation of these TAP compounds has made it possible to adjust the Q-band position in a stepwise manner between ca. 600 and 750 nm. The first reduction and oxidation potentials of the TAP ring shift negatively with increasing number and size of the fused aromatics. The extent of the shift is found to be very small for the LUMOs but significant for the HOMOs. These spectroscopic and electrochemical properties are almost perfectly reproduced by molecular orbital calculations within the framework of the Pariser-Parr-Pople approximation. In particular, a small variation of the LUMO level and large destabilization of the HOMO level on ring expansion are rationalized from the extent of stretch of molecular orbitals: i.e., since the LUMOs are localized in the central TAP moiety irrespective of the molecular size, while the HOMOs have appreciable coefficients even over the fused aromatics, the HOMO level destabilizes while the LUMO level remains constant with increasing molecular size. In one CoTAP derivative, Co(III/II) and the first ligand oxidation couples occur experimentally at the same potential. PMID- 12095347 TI - Octaphenylbiphenylene and dodecaphenyltriptycene. AB - Octaphenylbiphenylene, the expected dimer of tetraphenylbenzyne, has been prepared in low yield by diazotization of 3,4,5,6-tetraphenylanthranilic acid, and its X-ray structure has been determined. The X-ray structure of a second, abnormal dimer of tetraphenylbenzyne, 1,2,3,8,9,10 hexaphenyldibenzo[fg,op]naphthacene has also been determined; this is a saddle shaped polycyclic aromatic hydrocarbon. 1,2,3,4,5,6,7,8,13,14,15,16 Dodecaphenyltriptycene, perhaps the most crowded triptycene derivative yet prepared, has been made by the reaction of tetraphenylbenzyne with 1,2,3,4,5,6,7,8-octaphenylanthracene, which in turn was synthesized in two steps from commercial starting materials. The X-ray structure of the dodecaphenyltriptycene nonabenzene solvate is a remarkable channel containing structure in which more than 50% of the unit cell volume is occupied by the benzene molecules. PMID- 12095348 TI - Comparison of the binding of cadmium(II), mercury(II), and arsenic(III) to the de novo designed peptides TRI L12C and TRI L16C. AB - Designed alpha-helical peptides of the TRI family with a general sequence Ac G(LKALEEK)(4)G-CONH(2) were used as model systems for the study of metal-protein interactions. Variants containing cysteine residues in positions 12 (TRI L12C) and 16 (TRI L16C) were used for the metal binding studies. Cd(II) binding was investigated, and the results were compared with previous and current work on Hg(II) and As(III) binding. The metal peptide assemblies were studied with the use of UV, CD, EXAFS, (113)Cd NMR, and (111m)Cd perturbed angular correlation spectroscopy. The metalated peptide aggregates exhibited pH-dependent behavior. At high pH values, Cd(II) was bound to the three sulfurs of the three-stranded alpha-helical coiled coils. A mixture of two species was observed, including Cd(II) in a trigonal planar geometry. The complexes have UV bands at 231 nm (20 600 M(-1) cm(-1)) for TRI L12C and 232 nm (22 600 M(-1) cm(-1)) for TRI L16C, an average Cd-S bond length of 2.49 A for both cases, and a (113)Cd NMR chemical shift at 619 ppm (Cd(II)(TRI L12C)(3)(-)) or 625 ppm (Cd(II)(TRI-L16C)(3)(-)). Nuclear quadrupole interactions show that two different Cd species are present for both peptides. One species with omega(0) = 0.45 rad/ns and low eta is attributed to a trigonal planar Cd-(Cys)(3) site. The other, with a smaller omega(0), is attributed to a four-coordinate Cd(Cys)(3)(H(2)O) species. At low pH, no metal binding was observed. Hg(II) binding to TRI L12C was also found to be pH dependent, and a 3:1 sulfur-to-mercury(II) species was observed at pH 9.4. These metal peptide complexes provide insight into heavy metal binding and metalloregulatory proteins such as MerR or CadC. PMID- 12095349 TI - Targeted strand scission of DNA substrates by a tricopper(II) coordination complex. AB - A trinuclear copper complex, [Cu(3)(II)(L)(H(2)O)(3)(NO(3))(2)](NO(3))(4).5H(2)O (1) (L = 2,2',2' '-tris(dipicolylamino)triethylamine), with pyridyl and alkylamine coordination exhibits a remarkable ability to promote specific strand scission at junctions between single- and double-stranded DNA. Strand scission occurs on the 3' overhang at the junction of a hairpin or frayed duplex structure and is not dependent on the identity of the base at which cleavage occurs. Target recognition minimally requires a purine at the first unpaired position and a guanine at the second unpaired position on the 5' strand. Incorporation of the necessary recognition elements into an otherwise unreactive junction resulted in specific strand scission at that new target and helped to confirm the predictive nature of this complex. Selective strand scission requires both a reductant and dioxygen, suggesting activation of O(2) by the reduced form of 1. The reaction utilizing the trinuclear complex does not appear to involve a diffusible radical species as suggested by its high specificity of target oxidation and its lack of sensitivity to radical quenching agents. Comparisons between the trinuclear copper complex, mononuclear analogues of 1, and [Cu(OP)(2)](2+) (OP = 1,10 phenanthroline) indicate that recognition and reactivity described in this report are dependent on the multiple metal ions within the same complex which together support its unique activity. PMID- 12095350 TI - Exciplex intermediates in photoinduced electron transfer of porphyrin-fullerene dyads. AB - The photoinduced electron transfer in differently linked zinc porphyrin-fullerene dyads and their free-base porphyrin analogues was studied in polar and nonpolar solvents with femto- to nanosecond absorption and emission spectroscopies. A new intermediate state, different from the locally excited (LE) chromophores and the complete charge-separated (CCS) state, was observed. It was identified as an exciplex. The exciplex preceded the CCS state in polar benzonitrile and the excited singlet state of fullerene in nonpolar toluene. The behavior of the dyads was modeled by using a common kinetic scheme involving equilibria between the exciplex and LE chromophores. The scheme is suitable for all the studied porphyrin-fullerene compounds. The rates of reaction steps depended on the type of linkage between the moieties. The scheme and Marcus theory were applied to calculate electronic couplings for sequential reactions, and consistent results were obtained. PMID- 12095351 TI - Gas-phase structure of (1,1,1,5,5,5-hexafluoro-2,4-pentanedionato)(eta(2)-1,5 cyclooctadiene)copper(I), Cu(1,5-cod)(hfac), an important precursor for vapor deposition of copper. AB - The molecular structure of Cu(1,5-cod)(hfac) in the gas phase has been determined by electron diffraction, restrained by parameters calculated ab initio (MP2/AE1 level) or using Density Functional Theory (BP86/AE1 level). The most stable structure is one in which one olefinic group of the cyclooctadiene ligand is coordinated to the square-planar copper atom [refined Cu-C distances 194.0(13) and 194.4(9) pm]. The second C=C double bond is weakly associated with the copper atom [Cu...C distances 267.2(23) and 276.9(25) pm], and the cyclooctadiene ligand has a twist-boat conformation, so that the complex has C(1) symmetry. The nature of the bonding between copper and each of the two olefin moieties has been assessed by topological analysis of the BP86/AE1 total electron density. A form with C(2) symmetry, lying between 2 and 7 kJ mol(-1) above the ground state, is a transition state for exchange of the two olefinic groups. There are also two higher energy conformers, both 10 kJ mol(-1) or more above the ground state. In one of these the cyclooctadiene ligand retains the twist-boat conformation, but the Cu(hfac) moiety is coordinated in the exo position with respect to the noncoordinated olefin, instead of endo, as in the most stable conformer. The molecular symmetry is C(1) in this isomer. In the remaining form the ligand has the chair conformation, and the molecular symmetry is C(s). PMID- 12095353 TI - A (13)C INADEQUATE and HF-GIAO study of C(60)H(2) and C(60)H(6) identification of ring currents in a 1,2-dihydrofullerene. AB - The hydrofullerenes C(60)H(2) (1) and C(60)H(6) (2) have been prepared in (13)C enriched form and 2D INADEQUATE NMR spectra were measured. These spectra have provided unambiguous (13)C assignments for 2, and nearly unambiguous assignments for 1. In both cases, the most downfield resonances are immediately adjacent to the sp(3) carbons, despite the fact that these carbons are the least pyramidalized carbons in the molecule. Typically, (13)C chemical shifts move downfield with increasing pyramidalization (THETA(p)), but in these systems there is no strong correlation between THETA(p) and delta. HF-GIAO calculations are able to predict the chemical shifts, but provide little chemical insight into the origin of these chemical shifts. London theory reveals a significant paramagnetic ring current in 1, a feature that helps explain the (1)H shifts in these compounds and may contribute to the (13)C chemical shifts as well. PMID- 12095352 TI - On the Debye-Waller factor of hexagonal ice: a computer simulation study. AB - We investigate by molecular dynamics (MD) simulations the temperature dependence of the Debye-Waller (DW) factor of hexagonal ice with 25 different proton disordered configurations. Each initial configuration is composed of 288 water molecules with no net dipole moment. The intermolecular interaction of water is described by TIP4P potential. Each production run of the simulation is 15 ns or longer. We observe a change in slope of the DW factor around 200 K, which cannot be explained within the framework of either classical or quantum harmonic approximation. Configurations generated by MD simulations are subjected to the steepest descent energy minimization. Analysis of the local energy minimum structures reveals that water molecules above 200 K jump to other lattice sites via some local energy minimum structures which contain some water molecules sitting on the locations other than the lattice sites. As time evolves, these defect molecules move back and forth to the lattice sites yielding defect-free structures. Those motions are responsible for the unusual increase in the DW factor at high temperatures. In making a transition from an energy-minimum structure to another one, a small number of water molecules are involved in a highly cooperative fashion. The larger DW factor at higher temperature arises from jump-like motions of water molecules among these locally stable configurations which may or may not be a family of the proton-disordered ice forms satisfying the "ice rule". PMID- 12095354 TI - Self-oscillation of polymer chains with rhythmical soluble-insoluble changes. AB - Self-oscillation of polymer chains in an aqueous solution has been achieved. The ruthenium catalyst for the Belousov-Zhabotinsky reaction was polymerized by using N-isopropylacrylamide and dissolved into the solution containing the BZ substrates. Periodical soluble-insoluble changes of the polymer chain were spontaneously induced by the BZ reaction. The conformational oscillations of the polymer were measured as the optical transmittance changes of the solution. This is the first report that rhythmical and reversible soluble-insoluble changes of polymer chains are realized under constant and homogeneous conditions. The transducing system from chemical energy of the BZ reaction to optical information has been constructed. PMID- 12095355 TI - Do nonplanar distortions of porphyrins bring about strongly red-shifted electronic spectra? Controversy, consensus, new developments, and relevance to chelatases. AB - TDDFT calculations confirm a controversial proposal by DiMagno and co-workers that ruffling distortions, by themselves, do not bring about sizable red shifts in the electronic absorption spectra of "simple" nontransition-metal porphyrins. We now report that the same also holds for saddling distortions. The situation is more complex for transition metal porphyrins. For example, ruffling does bring about strong red shifts in the electronic spectra of nickel porphyrins because of a specific metal(d)-porphyrin(pi) orbital interaction. PMID- 12095356 TI - Electronic absorption, resonance Raman, and electrochemical studies of planar and saddled copper(III) meso-triarylcorroles. Highly substituent-sensitive Soret bands as a distinctive feature of high-valent transition metal corroles. AB - We present here a first systematic study of substituent effects in metallocorroles, based on electronic absorption, resonance Raman (RR), and infrared (IR) spectroscopic studies and electrochemical measurements on 10 copper(III) meso-triarylcorroles, Cu(III)[beta-Y(8)TArC], where the beta substituent Y = H or Br and the meso-aryl group Ar = C(6)F(5) or p-X-C(6)H(4) and X = CF(3), H, CH(3), and OCH(3). The results afford a number of significant inisights. (1) The RR (and IR) results show that at least two and possibly more high-frequency bands in the 1400-1550 cm(-1) region exhibit significant frequency downshifts on beta-octabromination and, thus, qualify as structure-sensitive marker bands. DFT geometry optimizations indicate that the saddled conformation should be clearly preferred for the beta-octabromo-meso-triarylcorrole derivatives studied and that beta-octabromination results in expansion of a number of skeletal bond distances of the corrole macrocycle, consistent with observed frequency downshifts. (2) Electrochemical measurements on planar Cu(III)[TArC] derivatives have shown that the para substituents on the meso-aryl groups exert a strong influence on the half-wave potentials for oxidation (rho(ox) = DeltaE(1/2ox)/Delta(3sigma) = 95 mV), suggesting that oxidation involves removal of an electron from the corrole "b(1)" HOMO, which has significant amplitudes at the meso postions and crudely resembles a porphyrin a(2u) HOMO in shape. In contrast, the Hammett rho(ox) is much lower for the nonplanar Cu(III)[Br(8)TArC] derivatives and we suggest that this ultimately results from a b(1)-to-a(2) HOMO reversal which in turn stems from a metal (d(x2 y2)-corrole ("b(1)") orbital interaction that becomes symmetry-allowed under a saddle distortion of the corrole macrocycle. In contrast to what has been observed for metallotetraphenylporphyrins, beta-octabromination dramatically raises the half-wave potential for one-electron oxidation of the triarylcorrole derivatives studied. This appears to be due to the fact that both the "a(2)" and "b(1)" HOMOs of a corrole (in C(2v) notation) have significantly higher amplitudes at the beta positions, compared to a porphyrin a(2u) HOMO. Thus, although many metallocorroles are significantly more easily oxidizable than analogous metalloporphyrins, certain beta-octahalogeno-meso-triarylcorrole derivatives can indeed be extremely electron deficient and oxidation resistant and may, therefore, find use as rugged catalysts or reagents under highly oxidizing conditions. (3) Finally, the Soret absorption maxima of high-valent metallotriarylcorroles exhibit a uniquely sensitive dependence on the substituents on the meso-aryl groups. Thus, on going from Cu(III)[T(p-CF(3)-P)C] (T(p-CF(3)-P)C = meso-tris((p-trifluoromethyl)phenyl)corrolato) to Cu(III)[T(p-OM P)C] (T(p-OM-P)C = meso-tris(p-methoxyphenyl)corrolato), the Soret maximum red shifts by 26 nm, from 407 to 433 nm. Similarly, on going from Cu(III)[Br(8)T(p CF(3)-P)C] (Br(8)T(p-CF(3)-P)C = beta-octabromo-meso-tris((p trifluoromethyl)phenyl)corrolato) to Cu(III)[Br(8)T(p-OM-P)C] (Br(8)T(p-CF(3)-P)C = beta-octabromo-meso-tris(p-methoxyphenyl)corrolato), the Soret maximum red shifts by 34 nm, from 434 to 468 nm. Time-dependent DFT calculations suggest that this substituent dependence reflects significant ligand-to-metal charge-transfer character of certain transitions in the Soret region. The optical spectra of free base and non-high-valent transition metal tetrapyrroles, in general, do not exhibit a similar substituent dependence. PMID- 12095357 TI - Electronic structure of high-valent transition metal corrolazine complexes. The young and innocent? AB - This is a first quantum chemical study of corrolazine complexes. DFT calculations suggest that despite their extremely contracted central cavities, compared with porphyrins, a variety of corrolazine complexes may be expected to exist as stable compounds. The calculations also indicate that corrolazine complexes may be regarded as strongly electron-deficient analogues of corrole complexes. Thus, the calculated valence ionization potentials of P(V) and Cu(III) corrolazine derivatives are dramatically higher than those of analogous corrole derivatives. In addition, DFT calculations on Fe(IV) and Mn(IV) corrole and corrolazine derivatives suggest that compared with the often noninnocent corrole ligands, corrolazines are electronically more innocent and stabilize "purer" high-valent states of transition metal ions. PMID- 12095358 TI - Symmetry-breaking phenomena in metalloporphyrin pi-cation radicals. AB - Density functional theory (DFT) calculations of the energetics, molecular structures, and spin density profiles of metalloporphyrin pi-cation radicals suggest that the common practice of describing these radicals in terms of a universal A(1u)/A(2u) dichotomy is often not justified, confirming a possibility first foreseen by Prendergast and Spiro (ref 15) over a decade ago on the basis of vibrational spectroscopy and semiempirical calculations. Because of near degeneracy of the a(1u) and a(2u) HOMOs of many metalloporphyrins, the cation radicals derived from these compounds undergo a pseudo-Jahn-Teller (pJT) distortion and are, therefore, best described as (2)A(u) with reference to the C(4h) point group, rather than as (2)A(1u) (D(4h) or (2)A(2u) (D(4h)). We find that the porphyrin cation radicals undergo a pJT distortion if the energy difference between the (2)A(1u) and (2)A(2u) pi-cation radicals, optimized under D(4h) symmetry constraints, is less than 0.15 eV. According to this criterion, metallo-porphine and metallo-OEP pi-cation radicals should always be pJT distorted and metallo-meso-tetrahalogenoporphyrin radicals should not. For [Zn(TPP(*))](+), the (2)A(1u)/(2)A(2u) energy difference is almost exactly at the threshold of 0.15 eV, consistent with the experimental observation of both symmetry-broken and undistorted structures for this species. The (2)A(1u)/(2)A(2u) energy difference (when the molecular geometries are optimized under a D(4h) symmetry constraint) also appears to govern whether the real pJT distorted cation radical is more A(1u)- or A(2u)-like in terms of its spin density profile. Because many metalloporphyrin pi-cation radicals exist as cofacial dimers in the crystalline phase, we examined the symmetries and structures of the model compounds [[Zn(P)](2)](+,2+) by means of DFT geometry optimizations. The results showed that dimerization has relatively little impact on the bond length alternation in the individual rings. A final interesting result, consistent with experiment, is that the bond length alternation in the delocalized mixed-valence dimer [[Zn(P)](2)](+) is about half that found for [[Zn(P)](2)](2+). PMID- 12095359 TI - Characterization of the interface dipole at organic/ metal interfaces. AB - In organics-based (opto)electronic devices, the interface dipoles formed at the organic/metal interfaces play a key role in determining the barrier for charge (hole or electron) injection between the metal electrodes and the active organic layers. The origin of this dipole is rationalized here from the results of a joint experimental and theoretical study based on the interaction between acrylonitrile, a pi-conjugated molecule, and transition metal surfaces (Cu, Ni, and Fe). The adsorption of acrylonitrile on these surfaces is investigated experimentally by photoelectron spectroscopies, while quantum mechanical methods based on density functional theory are used to study the systems theoretically. It appears that the interface dipole formed at an organic/metal interface can be divided into two contributions: (i) the first corresponds to the "chemical" dipole induced by a partial charge transfer between the organic layers and the metal upon chemisorption of the organic molecules on the metal surface, and (ii) the second relates to the change in metal surface dipole because of the modification of the metal electron density tail that is induced by the presence of the adsorbed organic molecules. Our analysis shows that the charge injection barrier in devices can be tuned by modulating various parameters: the chemical potential of the bare metal (given by its work function), the metal surface dipole, and the ionization potential and electron affinity of the organic layer. PMID- 12095360 TI - The elusive oxidant species of cytochrome P450 enzymes: characterization by combined quantum mechanical/molecular mechanical (QM/MM) calculations. AB - The primary oxidant of cytochrome P450 enzymes, Compound I, is hard to detect experimentally; in the case of cytochrome P450(cam), this intermediate does not accumulate in solution during the catalytic cycle even at temperatures as low as 200 K (ref 4). Theory can play an important role in characterizing such elusive species. We present here combined quantum mechanical/molecular mechanical (QM/MM) calculations of Compound I of cytochrome P450(cam) in the full enzyme environment as well as density functional studies of the isolated QM region. The calculations assign the ground state of the species, quantify the effect of polarization and hydrogen bonding on its properties, and show that the protein environment and its specific hydrogen bonding to the cysteinate ligand are crucial for sustaining the Fe-S bond and for preventing the full oxidation of the sulfur. PMID- 12095361 TI - Electron-mediating Cu(A) centers in proteins: a comparative high field (1)H ENDOR study. AB - High field (W-band, 95 GHz) pulsed electron-nuclear double resonance (ENDOR) measurements were carried out on a number of proteins that contain the mixed valence, binuclear electron-mediating Cu(A) center. These include nitrous oxide reductase (N(2)OR), the recombinant water-soluble fragment of subunit II of Thermus thermophilus cytochrome c oxidase (COX) ba(3) (M160T9), its M160QT0 mutant, where the weak axial methionine ligand has been replaced by a glutamine, and the engineered "purple" azurin (purpAz). The three-dimensional (3-D) structures of these proteins, apart from the mutant, are known. The EPR spectra of all samples showed the presence of a mononuclear Cu(II) impurity with EPR characteristics of a type II copper. At W-band, the g( perpendicular) features of this center and of Cu(A) are well resolved, thus allowing us to obtain a clean Cu(A) ENDOR spectrum. The latter consists of two types of ENDOR signals. The first includes the signals of the four strongly coupled cysteine beta-protons, with isotropic hyperfine couplings, A(iso), in the 7-15 MHz range. The second group consists of weakly coupled protons with a primarily anisotropic character with A(zz) < 3 MHz. Orientation selective ENDOR spectra were collected for N(2)OR, M160QT0, and purpAz, and simulations of the cysteine beta-protons signals provided their isotropic and anisotropic hyperfine interactions. A linear correlation with a negative slope was found between the maximum A(iso) value of the beta-protons and the copper hyperfine interaction. Comparison of the best-fit anisotropic hyperfine parameters with those calculated from dipolar interactions extracted from the available 3-D structures sets limit to the sulfur spin densities. Similarly, the small coupling spectral region was simulated on the basis of the 3-D structures and compared with the experimental spectra. It was found that the width of the powder patterns of the weakly coupled protons recorded at g(perpendicular) is mainly determined by the histidine H(epsilon)(1) protons. Furthermore, the splitting in the outer wings of these powder patterns indicates differences in the positions of the imidazole rings relative to the Cu(2)S(2) core. Comparison of the spectral features of the weakly coupled protons of M160QT0 with those of the other investigated proteins shows that they are very similar to those of purpAz, where the Cu(A) center is the most symmetric, but the copper spin density and the H(epsilon)(1)-Cu distances are somewhat smaller. All proteins show the presence of a proton with a significantly negative A(iso) value which is assigned to an amide proton of one of the cysteines. The simulations of both strongly and weakly coupled protons, along with the known copper hyperfine couplings, were used to estimate and compare the spin density distribution in the various Cu(A) centers. The largest sulfur spin density was found in M160T9, and the lowest was found in purpAz. In addition, using the relation between the A(iso) values of the four cysteine beta-protons and the H-C-S-S dihedral angles, the relative contribution of the hyperconjugation mechanism to A(iso) was determined. The largest contribution was found for M160T9, and the lowest was found for purpAz. Possible correlations between the spin density distribution, structural features, and electron-transfer functionality are finally suggested. PMID- 12095362 TI - Accelerating rare reactive events by means of a finite electronic temperature. AB - The range of chemical problems that are directly accessible to first-principles molecular dynamics simulations based on density functional theory is extended with a novel method apt to accelerate rare reactive events. The introduction of a finite electronic temperature within the Mermin formalism leads to a lowering of chemical activation barriers and thus to an exponential enhancement of the rate at which these reactions are observed during a first-principles molecular dynamics simulation. The method presented here makes direct use of the intrinsic chemical information encoded in the electronic structure, and is therefore able to lower selectively chemically relevant activation energies even in systems where many competing low-energy pathways for conformational transitions or diffusive motions are present. The performance of this new approach is demonstrated for a series of prototypical chemical reactions in gas and in condensed phase. A typical acceleration that can be achieved is, for example, a factor of 10(5) for the cis-trans isomerization of peroxynitrous acid in aqueous solution at room temperature. PMID- 12095363 TI - Oligosilsesquioxanes as versatile building blocks for the preparation of self assembled thin films. AB - A self-assembly approach to the preparation of nanocomposite siliceous thin films by using oligosilsesquioxanes as building blocks is presented. Poly(styrene-4 sulfonate), PSS, and octa(3-aminopropyl)silsesquioxane, NSi8, were layer-by-layer (LbL) assembled onto planar substrates and polystyrene (PS) particles, thus forming composite multilayers. We have clarified the binding properties of NSi8 to PSS by examining the pH influence on film buildup by microelectrophoresis (zeta-potential) and quartz crystal microgravimetry (QCM). The regular growth of PSS/NSi8 multilayers on planar supports was confirmed by surface plasmon resonance (SPR) spectroscopy and QCM. By applying the LbL coating procedure to spherical templates, we prepared compact, microporous hollow silica spheres by calcining PS spheres coated with (poly(allylamine hydrochloride) (PAH)/PSS)(2)/(NSi8/PSS)(n) (n varying from 3 to 12), at 750 degrees C, because of sintering of the octameric clusters (NSi8). Hollow spheres derived from coatings with n = 3 drastically altered in size (relative to the template core), depending on the size of the PS particles used. The novelty of this method for the nanofabrication of siliceous films stems from the use of well-defined and discrete building blocks, such as NSi8, leading to homogeneous organic-silica composite films as well as individual siliceous particles of variable size and shape. PMID- 12095365 TI - A piece of my mind. Waiting. PMID- 12095368 TI - Cheaper HIV drugs for poor nations bring a new challenge: monitoring treatment. PMID- 12095370 TI - AIDS knowledge gap. PMID- 12095369 TI - MTCT-Plus program has two goals: end maternal HIV transmission + treat mothers. PMID- 12095372 TI - HIV prevention apathy? PMID- 12095374 TI - From the World Health Organization. Health: A pathway to sustainable development. PMID- 12095375 TI - Dignity-conserving care at the end of life. PMID- 12095377 TI - Loss of HIV-1-specific immunity during treatment interruption in 2 chronically infected patients. PMID- 12095378 TI - HIV seroprevalence of adult males incarcerated for a sexual offense in Rhode Island, 1994-1999. PMID- 12095379 TI - Safety and tolerability of intermittent rifampin/pyrazinamide for the treatment of latent tuberculosis infection in prisoners. PMID- 12095380 TI - Policies for exposure to bloodborne pathogens among US-based international voluntary medical organizations. PMID- 12095381 TI - Dual vs single protease inhibitor therapy following antiretroviral treatment failure: a randomized trial. AB - CONTEXT: Management of antiretroviral treatment failure in patients receiving protease inhibitor (PI)-containing regimens is a therapeutic challenge. OBJECTIVE: To assess whether adding a second PI improves antiviral efficacy of a 4-drug combination in patients with virologic failure while taking a PI containing regimen. DESIGN: Multicenter, randomized, 4-arm trial, double-blind and placebo-controlled for second PI, conducted between October 1998 and April 2000, for which there was a 24-week primary analysis with extension to 48 weeks. SETTING: Thirty-one participating AIDS (acquired immunodeficiency syndrome) Clinical Trials Units in the United States. PARTICIPANTS: A total of 481 human immunodeficiency virus (HIV)-infected persons with prior exposure to a maximum of 3 PIs and viral load above 1000 copies/mL. INTERVENTION: Selectively randomized assignment (per prior PI exposure) to saquinavir (n = 116); indinavir (n = 69); nelfinavir (n = 139); or placebo twice per day (n = 157); in combination with amprenavir, abacavir, efavirenz, and adefovir dipivoxil. MAIN OUTCOME MEASURES: Primary efficacy analysis involved the proportion with viral load below 200 copies/mL at 24 weeks. Other measures were changes in viral load and CD4 cell count from baseline, adverse events, and HIV drug susceptibility. RESULTS: Of 481 patients, 148 (31%) had a viral load below 200 copies/mL at week 24. The proportions of patients with a viral load below 200 copies/mL in the saquinavir, indinavir, nelfinavir, and placebo arms were 34% (40/116), 36% (25/69), 34% (47/139), and 23% (36/157), respectively. The proportion in the combined dual-PI arms was higher than in the amprenavir-plus-placebo arm (35% [112/324] vs 23% [36/157], respectively; P =.002). Overall, a higher proportion of nonnucleoside reverse transcriptase inhibitor (NNRTI)-naive patients had a viral load below 200 copies/mL compared with NNRTI-experienced patients (43% [115/270] vs 16% [33/211], respectively; P<.001). Baseline HIV-1 hypersusceptibility to efavirenz (< or = 0.4-fold difference in susceptibility compared with reference virus) was associated with suppression of viral load at 24 weeks to below 200 copies/mL (odds ratio [OR], 3.49; 95% confidence interval [CI], 1.62-7.33; P =.001), and more than 10-fold reduction in efavirenz susceptibility, with less likelihood of suppression at 24 weeks (OR, 0.28; 95% CI, 0.09-0.87; P =.03). CONCLUSIONS: In this study of antiretroviral-experienced patients with advanced immunodeficiency, viral load suppression to below 200 copies/mL was achieved in 31% of patients with regimens containing 4 or 5 new drugs. Use of 2 PIs, being naive to NNRTIs, and baseline hypersusceptibility to efavirenz were associated with a favorable outcome. PMID- 12095382 TI - Time trends in primary HIV-1 drug resistance among recently infected persons. AB - CONTEXT: Transmission of multiclass drug-resistant human immunodeficiency virus type 1 (HIV-1) may increase with wider use of antiretroviral therapy. OBJECTIVE: To determine trends in prevalence of HIV-1 drug resistance among recently infected individuals in a geographic area with a high penetration of antiviral treatment. DESIGN, SETTING, AND PATIENTS: Consecutive case series of 225 patients referred to a San Francisco, Calif, hospital with recent HIV-1 infection from June 1996 through June 2001. MAIN OUTCOME MEASURE: Time trends in the prevalence of genotypic and phenotypic primary drug resistance. RESULTS: Mutations associated with resistance to nonnucleoside reverse transcriptase inhibitors (NNRTIs) steadily increased from 0% in 1996-1997 to 12 (13.2%) in 2000-2001 (P =.01). There was 1 mutation associated with protease inhibitor resistance in 1996 1997 (2.5%) and there were 7 (7.7%) in 2000-2001 (P =.25). Genotypic resistance to nucleoside reverse transcriptase inhibitors (NRTIs) initially decreased and then returned to prior levels (P =.007 for test of homogeneity). Genotypic resistance to 2 or more classes of drugs increased from 1 (2.5%) to 12 (13.2%) (P =.004), but only 1 infection (1.2%) in the latter period was resistant to all 3 classes of agents (P =.58). Primary phenotypic resistance decreased for NRTIs from 21% to 6.2% (P =.03) and increased for NNRTIs from 0 to 8 (9.9%) (P =.02). Phenotypic resistance increased for protease inhibitors from 2.6% to 6.2% (P =.32). Median time to virologic suppression (<500 copies/mL) during therapy was 12 weeks for patients with genotypic evidence of resistance compared with 5 weeks for patients with drug-sensitive infections (P =.02). CONCLUSIONS: The frequency of primary resistance to NNRTIs is increasing, although resistance to all available classes of antiretroviral therapy remains rare. Genotypic resistance testing in recently infected persons predicts time to viral suppression during therapy. PMID- 12095383 TI - Two-dose intrapartum/newborn nevirapine and standard antiretroviral therapy to reduce perinatal HIV transmission: a randomized trial. AB - CONTEXT: A 2-dose intrapartum/newborn nevirapine regimen reduced perinatal human immunodeficiency virus (HIV) transmission in Ugandan women not receiving antenatal antiretroviral therapy (ART). However, it is unknown whether the addition of the 2-dose nevirapine regimen to standard ART would further reduce perinatal HIV transmission. OBJECTIVE: To determine whether a 2-dose nevirapine regimen can decrease perinatal transmission of HIV in nonbreastfeeding women receiving standard ART. DESIGN AND SETTING: International, blinded, placebo controlled, phase 3 trial enrolling women between May 1997 and June 2000 at clinical sites providing care for HIV infection throughout the United States, Europe, Brazil, and the Bahamas. PARTICIPANTS: A total of 1270 women received nevirapine (n = 642) or placebo (n = 628). Infants were followed up for 6 months to determine HIV-infection status, which was available for 1248 deliveries. INTERVENTION: A 200-mg dose of oral nevirapine to women after onset of labor and a 2-mg/kg dose of oral nevirapine to newborns between 48 and 72 hours after birth. MAIN OUTCOME MEASURES: Detection of HIV infection in infants and grade 3 and 4 toxic effects in women and newborns. RESULTS: After review by the data and safety monitoring board, the trial was stopped early because the overall transmission rates were significantly lower than assumed for the study design. Antenatal ART included zidovudine alone in 23%; combinations without protease inhibitors in 36%; and combinations with protease inhibitors in 41%. Thirty-four percent of women had elective cesarean delivery. No significant safety concerns were identified for women or infants. Detection of HIV infection occurred in 9 (1.4%; 95% confidence interval [CI], 0.6%-2.7%) of 631 nevirapine group deliveries and 10 (1.6%; 95% CI, 0.8%-2.9%) of 617 placebo group deliveries. The 95% CI for the difference in transmission rate (-0.2) between the 2 study arms ranged from -1.5% in favor of nevirapine to 1.2% in favor of placebo (P =.82, Fisher exact test). The transmission rate was higher in women with lower baseline CD4 cell counts and higher delivery HIV RNA levels, but there was no significant difference between treatment arms in any subgroup. CONCLUSION: Risk of perinatal HIV transmission was low and no benefit from additional intrapartum/newborn nevirapine was demonstrated when women received prenatal care and antenatal ART, and elective cesarean section was made available. PMID- 12095384 TI - Hepatitis C and progression of HIV disease. AB - CONTEXT: Conflicting reports exist regarding the effect of hepatitis C virus (HCV) on the progression of human immunodeficiency virus (HIV) disease. OBJECTIVE: To assess the effect of HCV infection on clinical and immunologic progression of HIV disease and immunologic response to highly active antiretroviral therapy (HAART). DESIGN: Prospective cohort study. SETTING: University-based, urban HIV clinic in the United States. PATIENTS: There were 1955 patients enrolled between January 1995 and January 2001 who were eligible for analysis because of having at least 1 return visit to the clinic and being free of acquired immunodeficiency syndrome (AIDS) at enrollment. Median (interquartile range) length of follow-up was 2.19 (1.00-3.50) years for HCV infected and 2.00 (1.00-3.00) years for HCV-uninfected patients. MAIN OUTCOME MEASURES: Progression to an AIDS-defining illness, survival, and progression to a CD4 cell count below 200/microL; CD4 cell count change following initiation of effective HAART (resulting in a viral load of <400 copies/mL recorded at > or = 75% of measurements). RESULTS: No difference was detected in the risk of acquiring an AIDS-defining illness (HCV-infected patients, 231 events [26.4%] and HCV-uninfected patients, 264 events [24.4%]; relative hazard [RH], 1.03; 95% confidence interval [CI], 0.86-1.23) or in the risk of death (HCV-infected patients, 153 deaths [17.5%] and HCV-uninfected patients, 168 deaths [15.5%]; RH, 1.05; 95% CI, 0.85 -1.30). Although an increased risk of death was detected in the subgroup of 429 HCV-infected patients with a baseline CD4 cell count of 50/microL through 200/microL (RH, 1.51; 95% CI, 1.01-2.27), after adjustment for exposure to HAART and its effectiveness in a multivariate Cox regression analysis, death was not independently associated with HCV infection in this subgroup (RH, 1.01; 95% CI, 0.65-1.56). Similarly, in those receiving effective HAART (n = 208), there was no difference in the increase in CD4 cell count or CD4 percentage during HAART in HCV-infected compared with HCV-uninfected patients. CONCLUSIONS: Among patients in this urban US cohort, we did not detect evidence that HCV infection substantially alters the risk of dying, developing AIDS, or responding immunologically to HAART, especially after accounting for differences in its administration and effectiveness. PMID- 12095385 TI - Abacavir substitution for nucleoside analogs in patients with HIV lipoatrophy: a randomized trial. AB - CONTEXT: Peripheral lipoatrophy may complicate antiretroviral therapy of human immunodeficiency virus (HIV) infection, often related to duration and type of nucleoside analog therapy, and may have a mitochondrial pathogenesis. No proven therapy exists for lipoatrophy, but abacavir is a nucleoside analog that may be less toxic to mitochondria. OBJECTIVE: To determine if substitution of stavudine or zidovudine with abacavir improves HIV lipoatrophy without affecting control of HIV replication. DESIGN: Randomized, open-label 24-week study. SETTING: Seventeen hospital HIV outpatient clinics and primary care centers in Australia and England, with randomization from June 2000 through January 2001. PARTICIPANTS: A total of 111 adults (109 men) with moderate or severe lipoatrophy who were receiving stavudine (n = 85) or zidovudine (n = 26) and had stable plasma HIV RNA levels below 400 copies/mL and no prior abacavir therapy. INTERVENTION: Patients were randomly assigned to switch from stavudine or zidovudine to abacavir, 300 mg twice per day, while continuing all other antiretroviral therapy (n = 54) or to continue all antiretroviral therapy (n = 57). MAIN OUTCOME MEASURES: The primary end point was limb fat mass, measured by dual-energy x-ray absorptiometry; key secondary end points were plasma HIV RNA levels, adverse events, physician assessed (via subjective measures) lipodystrophy severity, total and central fat mass, and fasting metabolic (lipid, glycemic, and lactate) levels. RESULTS: There was a significant increase in limb fat in the abacavir group relative to the stavudine/zidovudine group (0.39 vs 0.08 kg; mean difference, 0.31; 95% confidence interval [CI], 0.06-0.57 kg), as well as significant relative increases in subcutaneous thigh (P =.01), arm (P<.001), and abdominal (P =.001) fat areas on computed tomography. Switching had no significant effect on secondary end points, including plasma HIV RNA (for unadjusted comparison between groups at week 24, odds ratio, 1.38; 95% CI, 0.48-3.96). Change in limb fat mass at week 24 did not correlate with change in subjectively determined perceived lipoatrophy severity (r = -0.06; P =.53 by Spearman correlation). Hypersensitivity to abacavir was seen in 5 patients (10%). CONCLUSIONS: In this sample of lipoatrophic HIV-infected adults, switching from stavudine or zidovudine to abacavir for 24 weeks led to significant, albeit modest, objectively measured increases in limb fat. Clinical lipoatrophy, as assessed subjectively, did not resolve, however, and at the rate of increase observed may take years to resolve with use of this strategy. Longer-term follow-up is needed. PMID- 12095386 TI - Real-time, universal screening for acute HIV infection in a routine HIV counseling and testing population. AB - CONTEXT: Acute human immunodeficiency virus (HIV) infection cannot be diagnosed by routine antibody tests and is rarely diagnosed in clinical practice. However, HIV nucleic acid-based testing is widely used to screen for antibody-negative acute infection among low-risk blood donors. OBJECTIVE: To assess the feasibility of screening in high-volume laboratories for acute and long-term HIV infection in a routine HIV testing population, in which HIV infection prevalence is low, using specimen pooling and HIV RNA reverse transcriptase-polymerase chain reaction (RT PCR) tests. DESIGN AND SETTING: Clinical diagnostic performance evaluation at a state-funded public health virology and serology laboratory. PARTICIPANTS: A total of 8505 consecutive individuals presenting for routine HIV counseling and testing during a total of 20 business days to simulate a month of testing in August and December 2001 at 110 publicly funded testing sites in North Carolina. MAIN OUTCOME MEASURES: Prevalence of acute and long-term HIV infection. Serum specimens negative by HIV enzyme immunoassay (EIA) were screened in pools by an ultrasensitive HIV RNA RT-PCR test. Results for individual HIV RNA-positive specimens were reclassified as true or false according to results of confirmatory testing. RESULTS: Of the 8505 individuals screened, 8194 had not previously tested HIV positive and had sufficient serum to complete the testing protocol. Of those, 39 had long-term HIV infection (prevalence, 47.6 per 10,000 at-risk persons [95% confidence interval, 33.8-65.0 per 10,000]). Of the 8155 at-risk individuals whose antibody tests were negative, 5 were HIV RNA positive. Four of those had true-positive acute infection (prevalence, 4.9 per 10,000 [95% confidence interval, 1.3-12.5 per 10,000]). All 4 were women; 2 developed symptoms consistent with an acute retroviral syndrome in the week after testing. Screening all specimens required 147 HIV RNA tests. Overall specificity of the strategy was 0.9999. CONCLUSIONS: These findings suggest the widespread diagnosis of acute HIV infections in a routine testing population is not only possible but feasible using specimen pooling and nucleic acid testing. These additional procedures may increase diagnostic yield by approximately 10% compared with conventional HIV antibody testing. PMID- 12095387 TI - Antiretroviral treatment for adult HIV infection in 2002: updated recommendations of the International AIDS Society-USA Panel. AB - OBJECTIVE: New information warrants updated recommendations for the 4 central issues in antiretroviral therapy: when to start, what drugs to start with, when to change, and what to change to. These updated recommendations are intended to guide practicing physicians actively involved in human immunodeficiency virus (HIV)- and acquired immunodeficiency syndrome (AIDS)-related care. PARTICIPANTS: In 1995, physicians with specific expertise in HIV-related basic science and clinical research, antiretroviral therapy, and HIV patient care were invited by the International AIDS Society-USA to serve on a volunteer panel. In 1999, others were invited to broaden international representation. The 17-member panel met regularly in closed meetings between its last report in 2000 and April 2002 to review current data. The effort was sponsored and funded by the International AIDS Society-USA, a not-for-profit physician education organization. EVIDENCE AND CONSENSUS PROCESS: The full panel was convened in late 2000 and assigned 7 section committees. A section writer and 3 to 5 section committee members (each panel member served on numerous sections) identified relevant evidence and prepared draft recommendations. Basic science, clinical research, and epidemiologic data from the published literature and abstracts from recent (within 2 years) scientific conferences were considered by strength of evidence. Extrapolations from basic science data and expert opinion of the panel members were included as evidence. Draft sections were combined and circulated to the entire panel and discussed in a series of full-panel conference calls until consensus was reached. Final recommendations represent full consensus agreement of the panel. CONCLUSIONS: Because of increased awareness of the activity and toxicity of current drugs, the threshold for initiation of therapy has shifted to a later time in the course of HIV disease. However, the optimal time to initiate therapy remains imprecisely defined. Availability of new drugs has broadened options for therapy initiation and management of treatment failure, which remains a difficult challenge. PMID- 12095388 TI - An unequal epidemic in an unequal world. PMID- 12095389 TI - Emerging resistance to nonnucleoside reverse transcriptase inhibitors: a warning and a challenge. PMID- 12095390 TI - Does hepatitis C virus infection increase the risk of HIV disease progression? PMID- 12095391 TI - Preventing mother-to-child HIV transmission: the first year of Thailand's national program. AB - CONTEXT: Each year in Thailand, about 10,000 children are born at risk for mother to-child human immunodeficiency virus (HIV) transmission. In 2000, Thailand implemented a national program to prevent mother-to-child HIV transmission. OBJECTIVE: To describe the results of implementation of the program. DESIGN: Monthly collection of summary data from hospitals. SETTING: Public health hospitals (n = 822) in all 12 regions of Thailand, representing 75 provinces, excluding Bangkok. PARTICIPANTS: Women giving birth from October 2000 through September 2001, including HIV-seropositive women and their neonates. MAIN OUTCOME MEASURES: Percentages of women giving birth who were tested for HIV, HIV seropositive women giving birth who received antenatal prophylactic antiretroviral drugs, and HIV-exposed neonates who received prophylactic antiretroviral drugs and infant formula. RESULTS: Among 573,655 women (range, 27,344-77,806 by region) giving birth, 554,912 (96.7%) received antenatal care (range, 91.9%-98.8% by region). Of 554,912 women giving birth who had antenatal care, 517,488 (93.3%) were tested for HIV (range, 87.7%-99.4% by region) before giving birth; of 18,743 women giving birth who did not have antenatal care, 13,314 (71.0%) were tested for HIV (range, 21.7%-92.9% by region). Of 6646 HIV seropositive women giving birth, 4659 (70.1%) received prophylactic antiretroviral drugs before delivery (range, 55.3%-81.2% by region). Of 6475 neonates of HIV-seropositive women, 5741 (88.7%) received prophylactic antiretroviral drugs (range, 67.4%-96.9% by region) and 5386 (83.2%) received infant formula (range, 65.3%-100% by region). CONCLUSIONS: Major program components of Thailand's national program for preventing mother-to-child HIV transmission were implemented. Thailand's experience may encourage other developing countries to implement or expand similar national programs. PMID- 12095397 TI - Effect of linsidomine on the human radial artery. AB - The response of the human radial artery to a direct NO donor, linsidomine or 3 morpholino-sydnonimine (Sin 1), in the therapeutic management of peri-operative spasm may increase the patency rates of these grafts in the short, medium and long term. Evaluation of the effects of Sin 1 on the human radial artery is of even greater interest as it has not been published previously. Ninety-six human radial artery rings were studied with two protocols. Rings were mounted in an isolated organ bath between two stainless steel metallic rods connected to stress gauges. Protocol 1 studied the vasorelaxant effect of Sin 1 and nitroglycerin (TNT). Protocol 2 studied the reactivity of the radial artery to the vasoconstrictor agents arginine vasopressin (AVP) and angiotensin II (ANG II). The vasorelaxant effect of Sin 1 on the human radial artery was comparable with that of TNT, but with no tolerance effect. After Sin 1 pre-incubation, the vasoconstrictor effect of ANG II was abolished, whereas AVP induced maximum vasoconstriction similar to that of the control (not statistically significant), but with a shift in the EC50 to higher concentrations, EC50=15+/-20 nM. Sin 1 vasorelaxation of rings precontracted by ANG II was maximal, whereas after contraction by AVP, relaxation remained less than 70%; Sin 1 is a potent vasorelaxant on the human radial artery, which does not exhibit cross-tolerance with nitrates. This compound may be used pre- or post-operatively, and would undoubtedly be of benefit in the peri-operative preparation bath. PMID- 12095398 TI - Empirical estimates of mean aortic pressure: advantages, drawbacks and implications for pressure redundancy. AB - Mean arterial pressure (MAP) is estimated at the brachial artery level by adding a fraction of pulse pressure (form factor; =0.33) to diastolic pressure. We tested the hypothesis that a fixed form factor can also be used at the aortic root level. We recorded systolic aortic pressure (SAP) and diastolic aortic pressure (DAP), and we calculated aortic pulse pressure (PP) and the time averaged MAP in the aorta of resting adults (n=73; age 43+/-14 years). Wave reflection was quantified using the augmentation index. The aortic form factor (range 0.35-0.53) decreased with age, MAP, PP and augmentation index (each P<0.001). The mean form factor value (0.45) gave a reasonable estimation of MAP (MAP=DAP+0.45PP; bias=0+/-2 mmHg), and the bias increased with MAP (P<0.001). An alternative formula (MAP=DAP+PP/3+5 mmHg) gave a more precise estimation (bias=0+/-1 mmHg), and the bias was not related to MAP. This latter formula was consistent with the previously reported mean pulse wave amplification of 15 mmHg, and with unchanged MAP and diastolic pressure from aorta to periphery. Multiple linear regression showed that 99% of the variability of MAP was explained by the combined influence of DAP and SAP, thus confirming major pressure redundancy. Results were obtained irrespective of whether the marked differences in heart period and extent of wave reflection between subjects were taken into account. In conclusion, the aortic form factor was strongly influenced by age, aortic pressure and wave reflection. An empirical formula (MAP=DAP+PP/3+5 mmHg) that is consistent with mechanical principles in the arterial system gave a more precise estimate of MAP in the aorta of resting humans. Only two distinct pressure powered functions were carried out in the (SAP, DAP, MAP, PP) four-pressure set. PMID- 12095399 TI - Increased nitric oxide elimination from the airways after smoking cessation. AB - Smokers have been found to have low exhaled nitric oxide (NO) levels. The aim of the present study was to investigate where in the respiratory system the decrease in NO occurs, and whether this decrease was affected by smoking cessation. Measurements of exhaled NO were carried out in smokers (n=20) and non-smoking control subjects (n=30). In nine of the smokers, exhaled NO was analysed 1, 2 and 4 weeks after smoking cessation. The level of exhaled NO at a flow rate of 0.1 litre/s was significantly lower in smokers (4+/-2 p.p.b.) than in non-smokers (7+/-5 p.p.b.; P=0.007). A calculation of the contributions from different areas of the lung showed that the NO flux from the airways was significantly lower (14+/-10 compared with 36+/-26 nl/min; P=0.0001) and the alveolar fraction was significantly higher (2.1+/-0.8 compared with 1.5+/-0.9 p.p.b.; P=0.006) in smokers than in non-smokers. Nine smoking subjects refrained from smoking for 4 weeks, and this resulted in increased NO flux from the airways of 28+/-17 nl/min, which was no longer significantly different from controls. In conclusion, endogenous production of NO in the airways is decreased in smokers, but can be restored to normal values by 4 weeks after cessation of smoking. Smokers have an increased alveolar fraction of NO, and this might be a diagnostic sign of lung damage. Thus NO monitoring can be used to indicate improvements when a smoker decides to stop smoking. PMID- 12095400 TI - Disruption of the elastin gene in adult Williams syndrome is accompanied by a paradoxical reduction in arterial stiffness. AB - Although the aetiology of Williams syndrome (WS) is related to elastin gene disruption, its pathogenesis remains unknown, particularly that of vascular lesions. The aim of the present study was to compare the elastic properties of three WS patients with age- and gender-matched normotensive and hypertensive controls. Common carotid arteries of WS patients had a higher distensibility, a thicker intima-media and a lower elastic modulus. Electron microscopy studies of one WS renal artery showed major abnormalities of the elastic fibres, which displayed a reticular structure and a thickening of the internal elastic lamina, whereas the ultrastructure of elastic fibres was normal in a control subadventitial muscular fibrodysplasia. In this WS arterial stenosis, we studied the expression patterns of several major smooth muscle (SM) phenotypic markers using immunofluorescence and used a normal renal artery as a control. In WS, SM alpha-actin- and myosin-heavy-chain-positive cells contained low amounts of heavy caldesmon, and laminin-beta1 chain was expressed into the basement membranes, indicating a less differentiated phenotype. In conclusion, in WS patients, the carotid artery wall was abnormally distensible and thick, and major ultrastructural abnormalities of elastic fibres were observed in association with smooth muscle cell de-differentiation. These results indicate that the haplo insufficiency of the elastin gene in WS patients leads to abnormal elastic fibre assembly within the media. Arterial wall hypertrophy found with a primary defect in elastin may represent a major factor responsible for increased distensibility. We suggest that, in WS, the increased proliferative response and the associated de-differentiation process represent two important mechanisms underlying the matrix accumulation and the development of arterial stenosis. PMID- 12095401 TI - Altered L-arginine metabolism results in increased nitric oxide release from uraemic endothelial cells. AB - Results regarding the nitric oxide (NO) system in uraemia are contradictory. L arginine, the precursor of NO, is also metabolized by arginase to form ornithine and urea. In the present study, endothelial NO production and arginine metabolism in uraemia were assessed. In addition an in vivo model was used to examine excess consumption of NO in uraemia. NO and amino acid measurements were made from basal and stimulated (by bradykinin) uraemic and control endothelial cells. Reverse transcriptase PCR was used to assess endothelial NO synthase (eNOS) and inducible NOS (iNOS) expression. Finally, aortae of uraemic rats were stained for nitrotyrosine (a marker of peroxynitrite). Basal uraemic cells produced more NO than the control cells. L-arginine levels were greater in uraemic (supernatants/cells), but ornithine levels were higher in control (supernatants/cells). Following stimulation, NO levels in supernatants were similar, but the rise in NO production was greater in control compared with uraemic cells; l-arginine levels still remained higher in uraemic supernatants/cells. Differences in ornithine concentration (supernatants/cells) disappeared following bradykinin stimulation, due to a rise in ornithine levels in the uraemic group. There was no difference in eNOS expression, nor was iNOS detected in either group. Only aortae from uraemic rats showed evidence for nitrotyrosine staining. These studies demonstrated increased basal NO release in uraemic endothelial cells, perhaps by inhibition of arginase and hence diversion of arginine to the NO pathway. The increased NO produced under basal conditions may be inactive due to excessive consumption, resulting in peroxynitrite formation. Interestingly, bradykinin appears to restore arginase activity in uraemia, resulting in normalization of NO production. PMID- 12095402 TI - Impaired sympathetic regulation of cerebral blood flow in patients with cirrhosis of the liver. AB - Continuous recording of mean cerebral blood flow velocity (MCBFV) by Doppler ultrasound allows detection of low-frequency (LF) oscillations, which reflect sympathetic activity in the cerebral circulation. To establish whether the sympathetic drive to the cerebral circulation is altered in patients with compensated cirrhosis, and, if so, where alterations take place, LF oscillations of MCBFV, heart rate (RR interval) and systolic arterial pressure (SAP) were analysed in 10 patients with cirrhosis and 10 control subjects during supine rest and on stimulation of carotid baroreceptors using a neck chamber applying sinusoidal suction. Bivariate analysis was used to study the relationship between pairs of oscillations. In the case of a significant association, the delay in the appearance of the oscillation in MCBFV, SAP and RR was calculated. Baroreceptor stimulation induced significant increases in SAP LF and RR LF power in both groups, while MCBFV LF power increased only in controls. During baroreceptor stimulation, the lag phase between SAP LF and MCBFV LF power was significantly lower in cirrhotic patients than in control subjects (0.96 compared with 1.59 rad; P<0.01), indicating altered sympathetic regulation of the cerebral circulation. The baroreflex arc was intact, as indicated by the similar pattern of RR-SAP interval in patients and controls. Plasma noradrenaline levels increased significantly in both groups in response to head-up tilt. These results indicate that patients with cirrhosis have an altered sympathetic regulation of the cerebral circulation that is characterized by an inadequate response of resistance microvessels, despite adequate baroreceptor function. PMID- 12095403 TI - Serum from healthy pregnant women reduces oxidative stress in human umbilical vein endothelial cells. AB - This study was conducted to compare the effects of serum from healthy pregnant women and that from pregnant women with pre-eclampsia on oxidative stress in endothelial cells in culture. Human umbilical vein endothelial cells (HUVECs) were incubated with serum from 18 pre-eclamptic, 18 healthy pregnant and 18 healthy non-pregnant women for 24 h. The levels of reduced glutathione (GSH) and lipid peroxides (LPOs) were measured in endothelial cell lysates. Measurement of malondialdehyde in combination with 4-hydroxyalkenals has been used as an indicator of LPOs. Serum from healthy pregnant women decreased significantly the LPO content in HUVECs in comparison with serum from pre-eclamptic women and healthy non-pregnant women (30.7+/-6.6 compared with 39.3+/-10.9 and 41.0+/-12.7 pmol/mg of protein respectively; P<0.003 and P<0.01 respectively). No differences in GSH content between the three groups (18.3+/-2.1 nmol/mg of protein for healthy pregnant, 19.2+/-3.3 nmol/mg for pre-eclamptic and 18.3+/-2.0 nmol/mg for healthy non-pregnant women) were found. Thus serum from normal pregnant women contains a factor(s) that decreases oxidative stress in human endothelial cells. This mechanism might be altered in pre-eclampsia. PMID- 12095404 TI - Reproducibility of derived central arterial waveforms in patients with chronic renal failure. AB - Arterial stiffness potently predicts mortality in dialysis patients. Pulse-wave analysis permits the non-invasive assessment of indices of arterial stiffness and the central pressure waveform by applanation tonometry. The aim of this study was to assess the reproducibility of pulse-wave analysis in patients with chronic renal failure. A total of 188 subjects (23 healthy controls, along with 71 pre dialysis, 67 dialysis and 27 transplant patients) took part. Duplicate measurements were recorded of brachial blood pressure using the semi-automated Omron 705 device and of the radial artery pressure waveform using applanation tonometry. The central pressure aortic waveform was then obtained by application of a transfer function incorporated into the SphygmoCor software. Central aortic mean blood pressure (MBP), indices of arterial stiffness [augmentation index (AIx) and time to reflection (TR)] and the subendocardial viability ratio (SEVR) were analysed for intra-observer, inter-observer and long-term reproducibility using Bland-Altman plots. The mean (+/-S.D.) intra-observer difference was 0+/-4% for AIx, 0+/-20 ms for TR, 0+/-3 mmHg for aortic MBP and 0+/-18% for the SEVR. Inter-observer mean differences were 0+/-3% for AIx, 1+/-7 ms for TR, 1+/-4 mmHg for aortic MBP and 1+/-9% for the SEVR. For the long-term study, the mean differences were -1+/-9% for AIx, -2+/-13 mmHg for aortic MBP, -2+/-12 ms for TR and 1+/-29% for the SEVR. Pulse-wave analysis showed excellent reproducibility in all the studies, and is therefore suitable for use in all patients with chronic renal failure. Further prospective and interventional studies are now required to assess whether AIx and TR are important prognostic indices of cardiovascular events, and therefore relevant surrogate indices of arterial stiffness in this susceptible population. PMID- 12095405 TI - Differential mechanisms of endothelium-dependent vasodilator responses in human myometrial small arteries in normal pregnancy and pre-eclampsia. AB - Pre-eclampsia is a pregnancy-specific disorder associated with hypertension and proteinuria, characterized by alterations in endothelial cell function. In the present study we have compared responses to the endothelium-dependent vasodilator, bradykinin, in small myometrial arteries from normal pregnant and non-pregnant women and women with pre-eclampsia, in order to assess the relative contributions of nitric oxide, endothelium-derived hyperpolarizing factor (EDHF) and prostanoids in mediating endothelium-dependent vasodilatation. Bradykinin induced concentration-dependent relaxation in arteries isolated from the three subject groups did not differ with regard to sensitivity or maximum response. Responses to bradykinin in all three groups were unaffected by cyclo-oxygenase inhibition alone, and were similarly unaffected by partial depolarization. The nitric oxide synthase (NOS) inhibitor, N-nitro-l-arginine methyl ester, significantly attenuated the responses to bradykinin in arteries from non pregnant women and almost abolished responses in arteries from women with pre eclampsia. However, in arteries from normal pregnant women, bradykinin-induced responses were maintained in the presence of NOS inhibition. Inhibition of NOS combined with partial depolarization abolished responses to bradykinin in these vessels. These results support the suggestion that, in the absence of NO, an EDHF can mediate vasodilator responses to bradykinin during normal pregnancy, an effect not apparent in arteries from non-pregnant women or women with pre eclampsia. The up-regulation of EDHF-type function may represent a vascular adaptation to normal pregnancy that is absent in pre-eclampsia, and this might contribute to the clinical features of the disease. PMID- 12095406 TI - Plasma brain natriuretic peptide as a predictor of haemodynamically significant patent ductus arteriosus in preterm infants. AB - Patent ductus arteriosus (PDA) is an important cause of morbidity in extremely preterm infants. As increased plasma brain natriuretic peptide (BNP) is a common feature of adult cardiac disease, we investigated the value of plasma BNP concentration as a predictor of haemodynamically significant PDA in very preterm infants. We studied 18 preterm infants (12 male) of median gestational age 30 weeks (range 24-34), median birth weight 1.46 kg (0.54-2.13) and 11 healthy term controls. Plasma BNP levels were measured by double-antibody radioimmunoassay on days 3, 5 and 7 of life, and an echocardiogram was performed on day 7. Six infants of median gestation 26 weeks (26-30), median birth weight 0.92 kg (0.54 1.04) had PDA proven by echocardiography on day 7. BNP concentrations (pg/ml) on day 3 were significantly higher in these infants than in the remaining twelve [median 2012 (786-2759) versus 42 (7-704), P<0.001]. In four infants PDA was treated successfully (one surgically, three with non-steroidal anti-inflammatory drugs). Two had haemodynamically insignificant closing ducts. In these infants with therapeutic or spontaneous resolution of a PDA, plasma BNP fell to normal values [median after treatment 9 pg/ml (8-27)]. Early measurement of plasma BNP in the first few days of life is a useful method for predicting those preterm infants who may require intervention for PDA. PMID- 12095407 TI - Natriuretic peptides as markers of patent ductus arteriosus in preterm infants. PMID- 12095408 TI - Measuring baroreflex sensitivity from the gain function between arterial pressure and heart period. AB - We tested an innovative approach for estimating baroreflex sensitivity (BRS) from the gain function between spontaneous oscillations of systolic arterial pressure (SAP) and heart period (HP). The major goal was to assess the practical implications of abandoning the classical coherence criterion (> or =0.5) as regards measurability of BRS, and agreement with values of BRS obtained using the phenylephrine test (Phe-BRS). We studied 19 normal subjects, 44 patients with a history of previous myocardial infarction (MI) and 45 patients with chronic heart failure (CHF). The experimental protocol included recording of SAP and HP for 10 min of supine rest, and evaluation of Phe-BRS. From resting SAP and HP, the gain and coherence functions were computed. The new BRS index was obtained in all subjects by averaging the gain function over the whole low-frequency band (0.04 0.15 Hz) (whole-band average BRS, WBA-BRS). WBA-BRS was 7.4 (5.8-10.8) ms/mmHg [median (25th-75th percentile)] in normal controls, 3.1 (1.4-5.4) ms/mmHg in MI patients (P<0.001 compared with normals) and 5.0 (3.2-6.9) ms/mmHg in CHF patients (P<0.01 compared with normals). Using the coherence criterion, BRS could be measured in only 43% and 49% of MI and CHF patients respectively, and the proportion of the low-frequency band contributing to the measurement was 21% (14 47%) and 29% (16-35%) respectively. The correlation between WBA-BRS and Phe-BRS was 0.47, 0.63 and 0.36 in the normal, MI and CHF groups respectively (all P<0.001). The relative bias of WBA-BRS was -5.2 ms/mmHg (P<0.001) in normals, 1.4 ms/mmHg (P=0.004) in MI patients and -1.0 ms/mmHg (P=0.11) in CHF patients. The limits of agreement were -13 to 2.6, -7.4 to 4.6 and -9.3 to 7.3 ms/mmHg in the normal, MI and CHF groups respectively. Thus the WBA-BRS method standardizes the computation of BRS among subjects, and dramatically increases its measurability in subjects with pathology compared with the classical spectral technique based on the coherence criterion. Compared with Phe-BRS, WBA-BRS tends to give negatively biased results. The correlation and the magnitude of the limits of agreement between the two methods are similar to those observed previously using coherence-based spectral methods. PMID- 12095409 TI - Assessing spontaneous baroreflex function: methodological considerations. PMID- 12095410 TI - Enhanced blood insulin overcomes pyruvate dehydrogenase derangements that reflect systemic insulin resistance in obese adolescents. AB - Pyruvate dehydrogenase (PDH) has low activity in the circulating lymphocytes (CL) of obese adolescents and adults. In vitro, it is unresponsive to insulin at 5 micro-units/ml and is activated at 50 micro-units/ml, in contrast with activation and inhibition respectively at these concentrations in CL from controls. These changes are seen as being indicative of a molecular disorder underlying insulin resistance. The aims of the present study were to determine whether a substantial enhancement of blood insulin levels restores the PDH activity in CL from obese adolescents and abolishes the in vitro alterations, and whether PDH activity and indices of insulin resistance are correlated. Six obese adolescents and six normal-weight controls underwent a 4 h frequently sampled intravenous glucose test with minimal model analysis, to bring about a sharp rise in blood insulin and provide a reliable index of insulin sensitivity (S(I)). PDH activity was evaluated in CL obtained from blood samples at set times before and after their exposure to insulin in vitro. Insulin levels rose in all subjects in the first 10 min, although to a much greater extent in the obese group, and then decreased until the end of the test (240 min; t(240)). PDH activity in CL paralleled the insulin pattern in the control subjects, whereas in the obese subjects it was below normal 3 min before the start of the test (t(-3)), but rose significantly throughout the test. PDH responses in vitro to insulin in CL taken from the control subjects at t(-3) and t(240) and in CL taken from the obese subjects at t(-3) were as reported above, but were normal (i.e. the same as in control CL) in CL taken from the obese subjects at t(240). Baseline PDH activity was inversely correlated with body mass index and with fasting insulin, and directly correlated with S(I). These results show that a brief and sharp enhancement of blood insulin overcomes derangements in PDH that reflect systemic insulin resistance in obese adolescents. PMID- 12095411 TI - Angiotensinogen M235T gene polymorphism and recurrent restenosis after repeated percutaneous transluminal coronary angiography. AB - The present study was designed to prospectively test the hypothesis that gene polymorphisms of the renin-angiotensin system are associated with recurrent restenosis after repeated percutaneous transluminal coronary angioplasty. Five hundred and eleven patients after first successful angioplasty were characterized with respect to the angiotensinogen M235T, angiotensin-converting enzyme insertion/deletion and angiotensin II type 1 receptor A1166C gene polymorphisms. In 164 of these patients repeated angioplasty on a restenotic lesion was performed. After repeated angioplasty, 46 patients had recurrent restenosis as defined by a greater than 50% progression of residual stenosis. In the recurrent restenosis group there was a statistically significant higher percentage of patients receiving cholesterol-lowering drugs compared with the group of patients without recurrent restenosis. The two groups of patients did not differ with respect to procedural and angiographic parameters. There were significantly more carriers of the angiotensinogen 235T allele in the recurrent restenosis group than in the control group without recurrent restenosis. No differences between the two groups were found with respect to the other gene polymorphisms investigated. According to the results of a multifactorial analysis of variance, only the 235T allele of the angiotensinogen gene and not cholesterol drug therapy independently affected the increase of stenosis at follow-up angiography. In conclusion, the angiotensinogen 235T allele may be an independent predictor for recurrent restenosis after repeated angioplasty. PMID- 12095412 TI - Is circulating nitrite a directly acting vasodilator? PMID- 12095414 TI - Expression of the leucocyte common antigen-related (LAR) tyrosine phosphatase is regulated by cell density through functional E-cadherin complexes. AB - The leucocyte common antigen-related phosphatase (LAR) has been implicated in receptor tyrosine kinase signalling pathways while also displaying cell-density dependency and localization to adherens junctions. Whereas physiological substrates for LAR have not been identified unequivocally, beta-catenin associates with LAR and is a substrate in vitro. With the implication that LAR may play a role in regulating E-cadherin-dependent cell-cell communication and contact inhibition, the relationship of LAR with E-cadherin was investigated. LAR expression increased with cell density in the human breast cancer cell line MCF-7 and in Ln 3 cells derived from the 13762NF rat mammary adenocarcinoma. LAR protein levels decreased rapidly when cells were replated at a low density after attaining high expression of LAR at high cell density. COS-7 cells displayed comparable density-dependent regulation of LAR expression when transiently expressing exogenous LAR under the control of a constitutively active promoter, indicating that the regulation of expression is not at the level of gene regulation. Disrupting homophilic E-cadherin complexes by chelating extracellular calcium caused a marked decrease in LAR protein levels. Similarly, blocking E cadherin interactions with saturating amounts of E-cadherin antibody (HECD-1) also led to a rapid and pronounced loss of cellular LAR. In contrast, mimicking cell-surface E-cadherin engagement by plating cells at low density on to dishes coated with HECD-1 resulted in a 2-fold increase in LAR expression compared with controls. These results suggest that density-dependent regulation of LAR expression is mediated by functional E-cadherin and may play a role in density dependent contact inhibition by regulating tyrosine phosphorylation in E-cadherin complexes. PMID- 12095415 TI - The metalloendopeptidase nardilysin (NRDc) is potently inhibited by heparin binding epidermal growth factor-like growth factor (HB-EGF). AB - Nardilysin (N-arginine dibasic convertase, or NRDc) is a cytosolic and cell surface metalloendopeptidase that, in vitro, cleaves substrates upstream of Arg or Lys in basic pairs. NRDc differs from most of the other members of the M16 family of metalloendopeptidases by a 90 amino acid acidic domain (DAC) inserted close to its active site. At the cell surface, NRDc binds heparin-binding epidermal growth factor-like growth factor (HB-EGF) and enhances HB-EGF-induced cell migration. An active-site mutant of NRDc fulfills this function as well as wild-type NRDc, indicating that the enzyme activity is not required for this process. We now demonstrate that NRDc starts at Met(49). Furthermore, we show that HB-EGF not only binds to NRDc but also potently inhibits its enzymic activity. NRDc-HB-EGF interaction involves the 21 amino acid heparin-binding domain (P21) of the growth factor, the DAC of NRDc and most probably its active site. Only disulphide-bonded P21 dimers are inhibitory. We also show that Ca(2+), via the DAC, regulates both NRDc activity and HB-EGF binding. We conclude that the DAC is thus a key regulatory element for the two distinct functions that NRDc fulfills, i.e. as an HB-EGF modulator and a peptidase. PMID- 12095416 TI - Rapid insertion of GLUT2 into the rat jejunal brush-border membrane promoted by glucagon-like peptide 2. AB - A possible role for GLUT2 transiently expressed in the rat jejunal brush-border membrane (BBM) under the influence of glucagon-like peptide 2 (GLP-2) was investigated using in vivo perfusion of the intestinal lumen as well as isolation of membrane proteins and immunohistochemistry. A 1 h vascular infusion of GLP-2 in vivo doubled the rate of fructose absorption and this increase could be blocked by luminal phloretin. Immunohistochemistry of frozen sections of rat jejunum showed the expression of GLUT2 in both the basolateral and BBMs of mature enterocytes. Perfusion of the intestinal lumen with 50 mM D-glucose or vascular infusion of 800 pM GLP-2 for 1 h increased the expression of GLUT2 in the BBM. Quantification of these changes using Western blotting of biotinylated surface exposed protein showed a doubling of the expression of GLUT2 in the BBM, but the effects of glucose and GLP-2 were not additive. These results indicate that vascular GLP-2 can promote the insertion of GLUT2 into the rat jejunal BBM providing a low-affinity/high-capacity route of entry for dietary hexoses. PMID- 12095417 TI - Genistein inhibits CCAAT/enhancer-binding protein beta (C/EBPbeta) activity and 3T3-L1 adipogenesis by increasing C/EBP homologous protein expression. AB - The tyrosine kinase inhibitor genistein inhibits 3T3-L1 adipogenesis when present during the first 72 h of differentiation. In this report, we investigated the underlying mechanisms involved in the anti-adipogenic effects of genistein. We found that genistein blocked the DNA binding and transcriptional activity of CCAAT/enhancer-binding protein beta (C/EBPbeta) during differentiation by promoting the expression of C/EBP homologous protein, a dominant-negative member of the C/EBP family. Loss of C/EBPbeta activity was manifested as a loss of differentiation-induced C/EBPalpha and peroxisome-proliferator-activated receptor gamma protein expression and a dramatic reduction in lipid accumulation. Further, we documented for the first time that C/EBPbeta was tyrosine-phosphorylated in vivo during differentiation and in vitro by activated epidermal growth factor receptor. Genistein inhibited both of these events. Collectively, these results indicate that genistein blocks adipogenesis and C/EBPbeta activity by increasing the level of C/EBP homologous protein and possibly by inhibiting the tyrosine phosphorylation of C/EBPbeta. PMID- 12095418 TI - Activation of macrophages by silicones: phenotype and production of oxidant metabolites. AB - BACKGROUND: The effect of silicones on the immune function is not fully characterized. In clinical and experimental studies, immune alterations associated with silicone gel seem to be related to macrophage activation. In this work we examined in vivo, phenotypic and functional changes on peritoneal macrophages early (24 h or 48 h) and late (45 days) after the intraperitoneal (i.p.) injection of dimethylpolysiloxane (DMPS) (silicone). We studied the expression of adhesion and co-stimulatory molecules and both the spontaneous and the stimulated production of reactive oxygen intermediates and nitric oxide (NO). RESULTS: The results presented here demonstrate that the fluid compound DMPS induced a persistent cell recruitment at the site of the injection. Besides, cell activation was still evident 45 days after the silicone injection: activated macrophages exhibited an increased expression of adhesion (CD54 and CD44) and co stimulatory molecules (CD86) and an enhanced production of oxidant metabolites and NO. CONCLUSIONS: Silicones induced a persistent recruitment of leukocytes at the site of the injection and macrophage activation was still evident 45 days after the injection. PMID- 12095419 TI - A transcriptional response to Wnt protein in human embryonic carcinoma cells. AB - BACKGROUND: Wnt signaling is implicated in many developmental decisions, including stem cell control, as well as in cancer. There are relatively few target genes known of the Wnt pathway. RESULTS: We have identified target genes of Wnt signaling using microarray technology and human embryonic carcinoma cells stimulated with active Wnt protein. The ~50 genes upregulated early after Wnt addition include the previously known Wnt targets Cyclin D1, MYC, ID2 and betaTRCP. The newly identified targets, which include MSX1, MSX2, Nucleophosmin, Follistatin, TLE/Groucho, Ubc4/5E2, CBP/P300, Frizzled and REST/NRSF, have important implications for understanding the roles of Wnts in development and cancer. The protein synthesis inhibitor cycloheximide blocks induction by Wnt, consistent with a requirement for newly synthesized beta-catenin protein prior to target gene activation. The promoters of nearly all the target genes we identified have putative TCF binding sites, and we show that the TCF binding site is required for induction of Follistatin. Several of the target genes have a cooperative response to a combination of Wnt and BMP. CONCLUSIONS: Wnt signaling activates genes that promote stem cell fate and inhibit cellular differentiation and regulates a remarkable number of genes involved in its own signaling system. PMID- 12095420 TI - Nucleolar localization of an isoform of the IGF-I precursor. AB - BACKGROUND Alternative exons encode different isoforms of the human insulin-like growth factor-I (IGF-I) precursor without altering mature IGF-I. We hypothesized that the various IGF-I precursors may traffic IGF-I differently. Chimeric IGF-I precursors were made with green fluorescent protein (GFP) cloned between the signal and mature IGF-I domains. RESULTS Chimeras containing exons 1 or 2 were located in the cytoplasm, consistent with a secretory pathway, and suggesting that both exons encoded functional signal peptides. Exon 5-containing chimeras localized to the nucleus and strongly to the nucleolus, while chimeras containing exon 6 or the upstream portion of exon 5 did not. Nuclear and nucleolar localization also occurred when the mature IGF-I domain was deleted from the chimeras, or when signal peptides were deleted. CONCLUSIONS We have identified a nucleolar localization for an isoform of the human IGF-I precursor. The findings are consistent with the presence of a nuclear and nucleolar localization signal situated in the C-terminal part of the exon 5-encoded domain with similarities to signals in several other growth factors. PMID- 12095421 TI - A memory-efficient dynamic programming algorithm for optimal alignment of a sequence to an RNA secondary structure. AB - BACKGROUND: Covariance models (CMs) are probabilistic models of RNA secondary structure, analogous to profile hidden Markov models of linear sequence. The dynamic programming algorithm for aligning a CM to an RNA sequence of length N is O(N3) in memory. This is only practical for small RNAs. RESULTS: I describe a divide and conquer variant of the alignment algorithm that is analogous to memory efficient Myers/Miller dynamic programming algorithms for linear sequence alignment. The new algorithm has an O(N2 log N) memory complexity, at the expense of a small constant factor in time. CONCLUSIONS: Optimal ribosomal RNA structural alignments that previously required up to 150 GB of memory now require less than 270 MB. PMID- 12095422 TI - The limit fold change model: a practical approach for selecting differentially expressed genes from microarray data. AB - BACKGROUND: The biomedical community is developing new methods of data analysis to more efficiently process the massive data sets produced by microarray experiments. Systematic and global mathematical approaches that can be readily applied to a large number of experimental designs become fundamental to correctly handle the otherwise overwhelming data sets. RESULTS: The gene selection model presented herein is based on the observation that: (1) variance of gene expression is a function of absolute expression; (2) one can model this relationship in order to set an appropriate lower fold change limit of significance; and (3) this relationship defines a function that can be used to select differentially expressed genes. The model first evaluates fold change (FC) across the entire range of absolute expression levels for any number of experimental conditions. Genes are systematically binned, and those genes within the top X% of highest FCs for each bin are evaluated both with and without the use of replicates. A function is fitted through the top X% of each bin, thereby defining a limit fold change. All genes selected by the 5% FC model lie above measurement variability using a within standard deviation (SDwithin) confidence level of 99.9%. Real time-PCR (RT-PCR) analysis demonstrated 85.7% concordance with microarray data selected by the limit function. CONCLUSION: The FC model can confidently select differentially expressed genes as corroborated by variance data and RT-PCR. The simplicity of the overall process permits selecting model limits that best describe experimental data by extracting information on gene expression patterns across the range of expression levels. Genes selected by this process can be consistently compared between experiments and enables the user to globally extract information with a high degree of confidence. PMID- 12095423 TI - The use of alternative therapies in the Saskatchewan stroke rehabilitation population. AB - BACKGROUND: Many patients use alternative therapies. The purpose of this study was to determine the percentage of stroke rehabilitation patients in Saskatchewan using alternative therapies, whether patients found these therapies effective in alleviating stroke-related symptoms, how often those patients who used alternative therapies discuss this fact with their primary care doctor and the main reason why patients might not do so. METHODS: Telephone questionnaire surveys were conducted with 117 patients who had suffered a stroke and undergone inpatient or outpatient rehabilitation at Saskatoon City Hospital. RESULTS: The study revealed that 26.5% of 117 stroke rehabilitation patients visited alternative practitioners at least once or used some form of unconventional therapy. Only 16.1% of patients found that alternative therapy made them feel much better. Of those who used alternative therapy, 61.3% did not discuss this fact with their primary physician. Many of the respondents (47.3%) who did not inform their physician stated that they did not see the necessity of talking about these treatments and 21.1% did not discuss the issue with their physician because they felt that he or she might disapprove of alternative therapies. CONCLUSION: A relatively small percentage of stroke patients found alternative therapies beneficial. Doctors should be aware that a significant number of patients will try alternative treatment without discussion with their primary care physician or specialist. The current study suggests that after completing routine questioning, doctors should also ask their patients about their use of alternative therapies and, when appropriate, review issues of safety and efficacy. PMID- 12095424 TI - Distribution of serotonergic and dopaminergic nerve fibers in the salivary gland complex of the cockroach Periplaneta americana. AB - BACKGROUND: The cockroach salivary gland consists of secretory acini with peripheral ion-transporting cells and central protein-producing cells, an extensive duct system, and a pair of reservoirs. Salivation is controlled by serotonergic and dopaminergic innervation. Serotonin stimulates the secretion of a protein-rich saliva, dopamine causes the production of a saliva without proteins. These findings suggest a model in which serotonin acts on the central cells and possibly other cell types, and dopamine acts selectively on the ion transporting cells. To examine this model, we have analyzed the spatial relationship of dopaminergic and serotonergic nerve fibers to the various cell types. RESULTS: The acinar tissue is entangled in a meshwork of serotonergic and dopaminergic varicose fibers. Dopaminergic fibers reside only at the surface of the acini next to the peripheral cells. Serotonergic fibers invade the acini and form a dense network between central cells. Salivary duct segments close to the acini are locally associated with dopaminergic and serotonergic fibers, whereas duct segments further downstream have only dopaminergic fibers on their surface and within the epithelium. In addition, the reservoirs have both a dopaminergic and a serotonergic innervation. CONCLUSION: Our results suggest that dopamine is released on the acinar surface, close to peripheral cells, and along the entire duct system. Serotonin is probably released close to peripheral and central cells, and at initial segments of the duct system. Moreover, the presence of serotonergic and dopaminergic fiber terminals on the reservoir indicates that the functions of this structure are also regulated by dopamine and serotonin. PMID- 12095425 TI - Preconceptional factors associated with very low birthweight delivery in East and West Berlin: a case control study. AB - BACKGROUND: Very low birthweight, i.e. a birthweight < 1500 g, is among the strongest determinants of infant mortality and childhood morbidity. To develop primary prevention approaches to VLBW birth and its sequelae, information is needed on the causes of preterm birth, their personal and social antecedents, and on conditions associated with very low birthweight. Despite the growing body of evidence linking sociodemographic variables with preterm delivery, little is known as to how this may be extrapolated to the risk of very low birthweight. METHODS: In 1992, two years after the German unification, we started to recruit two cohorts of very low birthweight infants and controls in East and West Berlin for a long-term neurodevelopmental study. The present analysis was undertaken to compare potential preconceptional risk factors for very low birthweight delivery in a case-control design including 166 mothers (82 East vs. 84 West Berlin) with very low birthweight delivery and 341 control mothers (166 East vs. 175 West). RESULTS: Multivariate logistic regression analysis was used to assess the effects of various dichotomous parental covariates and their interaction with living in East or West Berlin. After backward variable selection, short maternal school education, maternal unemployment, single-room apartment, smoking, previous preterm delivery, and fetal loss emerged as significant main effect variables, together with living in West Berlin as positive effect modificator for single mother status. CONCLUSION: Very low birthweight has been differentially associated with obstetrical history and indicators of maternal socioeconomic status in East and West Berlin. The ranking of these risk factors is under the influence of the political framework. PMID- 12095427 TI - Increased risk of traffic accidents in subjects with latent toxoplasmosis: a retrospective case-control study. AB - BACKGROUND: The parasite Toxoplasma gondii infects 30-60% of humans worldwide. Latent toxoplasmosis, i.e., the life-long presence of Toxoplasma cysts in neural and muscular tissues, leads to prolongation of reaction times in infected subjects. It is not know, however, whether the changes observed in laboratory influence the performance of subjects in real-life situations. METHODS: The seroprevalence of latent toxoplasmosis in subjects involved in traffic accidents (N=146) and in the general population living in the same area (N=446) was compared by a Mantel-Haenszel test for age-stratified data. Correlation between relative risk of traffic accident and a level of anti-Toxoplasma antibody titre was evaluated with the Cochran-Armitage test for trend. RESULTS: A higher seroprevalence was found in the traffic accident set than in the general population (Chi2MH=21.45, p<0.0001). The value of the odds ratio (OR) suggests that subjects with latent toxoplasmosis had a 2.65 (C.I.95= 1.764.01) times higher risk of an accident than the toxoplasmosis-negative subjects. The OR significantly increased with level of anti-Toxoplasma antibody titre (p<0.0001), being low (OR=1.86, C.I.95=1.14-3.03) for the 99 subjects with low antibody titres (8 and 16), higher (OR=4.78, C.I.95=2.39-9.59) for the 37 subjects with moderate titres (32 and 64), and very high (OR=16.03, C.I.95=1.89-135.66) for the 6 subjects with titres higher than 64. CONCLUSION: The subjects with latent toxoplasmosis have significantly increased risk of traffic accidents than the noninfected subjects. Relative risk of traffic accidents decreases with the duration of infection. These results suggest that 'asymptomatic' acquired toxoplasmosis might in fact represent a serious and highly underestimated public health as well as economic problem. PMID- 12095428 TI - Anthrax of the gastrointestinal tract. AB - When swallowed, anthrax spores may cause lesions from the oral cavity to the cecum. Gastrointestinal anthrax is greatly underreported in rural disease-endemic areas of the world. The apparent paucity of this form of anthrax reflects the lack of facilities able to make the diagnosis in these areas. The spectrum of disease, ranging from subclinical infection to death, has not been fully recognized. In some community-based studies, cases of gastrointestinal anthrax outnumbered those of cutaneous anthrax. The oropharyngeal variant, in particular, is unfamiliar to most physicians. The clinical features of oropharyngeal anthrax include fever and toxemia, inflammatory lesion(s) in the oral cavity or oropharynx, enlargement of cervical lymph nodes associated with edema of the soft tissue of the cervical area, and a high case-fatality rate. Awareness of gastrointestinal anthrax in a differential diagnosis remains important in anthrax endemic areas but now also in settings of possible bioterrorism. PMID- 12095429 TI - Emergence of Usutu virus, an African mosquito-borne flavivirus of the Japanese encephalitis virus group, central Europe. AB - During late summer 2001 in Austria, a series of deaths in several species of birds occurred, similar to the beginning of the West Nile virus (WNV) epidemic in the United States. We necropsied the dead birds and examined them by various methods; pathologic and immunohistologic investigations suggested a WNV infection. Subsequently, the virus was isolated, identified, partially sequenced, and subjected to phylogenetic analysis. The isolates exhibited 97% identity to Usutu virus (USUV), a mosquito-borne Flavivirus of the Japanese encephalitis virus group; USUV has never previously been observed outside Africa nor associated with fatal disease in animals or humans. If established in central Europe, this virus may have considerable effects on avian populations; whether USUV has the potential to cause severe human disease is unknown. PMID- 12095430 TI - First human isolate of Hantavirus (Andes virus) in the Americas. AB - We isolated Andes virus (formal name: Andes virus [ANDV], a species in the genus Hantavirus), from serum of an asymptomatic 10-year-old Chilean boy who died 6 days later of hantavirus pulmonary syndrome (HPS). The serum was obtained 12 days after his grandmother died from HPS and 2 days before he became febrile. No hantavirus immunoglobulin (Ig) G or IgM antibodies were detected in the serum sample. After three blind passages, ANDV antigens were detected in Vero E6 cells by immunofluorescence assay and enzyme-linked immunosorbent assay, and ANDV RNA was detected by reverse transcription-polymerase chain reaction. A fragment of the virus genome showed 96.2% nucleotide identity with that of prototype ANDV. To our knowledge, this is the first isolation of any agent of hemorrhagic fever with HPS from a human and the first such isolation of hantavirus before symptoms of that syndrome or HPS began. PMID- 12095431 TI - Ecologic niche modeling and potential reservoirs for Chagas disease, Mexico. AB - Ecologic niche modeling may improve our understanding of epidemiologically relevant vector and parasite-reservoir distributions. We used this tool to identify host relationships of Triatoma species implicated in transmission of Chagas disease. Associations have been documented between the protracta complex (Triatoma: Triatominae: Reduviidae) with packrat species (Neotoma spp.), providing an excellent case study for the broader challenge of developing hypotheses of association. Species pairs that were identified coincided exactly with those in previous studies, suggesting that local interactions between Triatoma and Neotoma species and subspecies have implications at a geographic level. Nothing is known about sylvatic associates of T. barberi, which are considered the primary Chagas vector in Mexico; its geographic distribution coincided closely with that of N. mexicana, suggesting interaction. The presence of this species was confirmed in two regions where it had been predicted but not previously collected. This approach may help in identifying Chagas disease risk areas, planning vector-control strategies, and exploring parasite-reservoir associations for other emerging diseases. PMID- 12095432 TI - Ehrlichia ewingii infection in white-tailed deer (Odocoileus virginianus). AB - Two closely related zoonotic ehrlichiae, Ehrlichia chaffeensis and E. ewingii, are transmitted by Amblyomma americanum, the lone star tick. Because white-tailed deer (Odocoileus virginianus) are critical hosts for all mobile stages of A. americanum and are important vertebrate reservoirs of E. chaffeensis, we investigated whether deer may be infected with E. ewingii, a cause of granulocytotropic ehrlichiosis in humans and dogs. To test for E. ewingii infection, we used polymerase chain reaction and inoculation of fawns with whole blood from wild deer. Of 110 deer tested from 20 locations in 8 U.S. states, 6 (5.5%) were positive for E. ewingii. In addition, natural E. ewingii infection was confirmed through infection of captive fawns. These findings expand the geographic distribution of E. ewingii, along with risk for human infection, to include areas of Kentucky, Georgia, and South Carolina. These data suggest that white-tailed deer may be an important reservoir for E. ewingii. PMID- 12095426 TI - The glial growth factors deficiency and synaptic destabilization hypothesis of schizophrenia. AB - BACKGROUND: A systems approach to understanding the etiology of schizophrenia requires a theory which is able to integrate genetic as well as neurodevelopmental factors. PRESENTATION OF THE HYPOTHESIS: Based on a co localization of loci approach and a large amount of circumstantial evidence, we here propose that a functional deficiency of glial growth factors and of growth factors produced by glial cells are among the distal causes in the genotype-to phenotype chain leading to the development of schizophrenia. These factors include neuregulin, insulin-like growth factor I, insulin, epidermal growth factor, neurotrophic growth factors, erbB receptors, phosphatidylinositol-3 kinase, growth arrest specific genes, neuritin, tumor necrosis factor alpha, glutamate, NMDA and cholinergic receptors. A genetically and epigenetically determined low baseline of glial growth factor signaling and synaptic strength is expected to increase the vulnerability for additional reductions (e.g., by viruses such as HHV-6 and JC virus infecting glial cells). This should lead to a weakening of the positive feedback loop between the presynaptic neuron and its targets, and below a certain threshold to synaptic destabilization and schizophrenia. TESTING THE HYPOTHESIS: Supported by informed conjectures and empirical facts, the hypothesis makes an attractive case for a large number of further investigations. IMPLICATIONS OF THE HYPOTHESIS: The hypothesis suggests glial cells as the locus of the genes-environment interactions in schizophrenia, with glial asthenia as an important factor for the genetic liability to the disorder, and an increase of prolactin and/or insulin as possible working mechanisms of traditional and atypical neuroleptic treatments. PMID- 12095433 TI - Time-space clustering of human brucellosis, California, 1973-1992. AB - Infection with Brucella spp. continues to pose a human health risk in California despite great strides in eradicating the disease from domestic animals. Clustering of human cases in time and space has important public health implications for understanding risk factors and sources of infection. Temporal spatial clustering of human brucellosis in California for the 20-year period 1973 1992 was evaluated by the Ederer-Myers-Mantel, Moran's I, and population-adjusted Moran's I procedures. Cases were clustered in concentrated agricultural regions in the first 5-year interval (1973-1977). Time-space clustering of human brucellosis cases in California late in the 20-year study period may reflect the distribution of Hispanic populations. Public health programs in California should focus on educating Hispanic populations about the risk of consuming dairy products, such as soft cheeses, made from unpasteurized milk. PMID- 12095434 TI - Persistent high incidence of tuberculosis in immigrants in a low-incidence country. AB - Immigration from areas of high incidence is thought to have fueled the resurgence of tuberculosis (TB) in areas of low incidence. To reduce the risk of disease in low-incidence areas, the main countermeasure has been the screening of immigrants on arrival. This measure is based on the assumption of a prompt decline in the incidence of TB in immigrants during their first few years of residence in a country with low overall incidence. We have documented that this assumption is not true for 619 Somali immigrants reported in Denmark as having TB. The annual incidence of TB declined only gradually during the first 7 years of residence, from an initial 2,000 per 100,000 to 700 per 100,000. The decline was described by an exponential function with a half-time of 5.7 (95% confidence interval 4.0 to 9.7) years. This finding seriously challenges the adequacy of the customary practice of screening solely on arrival. PMID- 12095435 TI - Automatic electronic laboratory-based reporting of notifiable infectious diseases at a large health system. AB - Electronic laboratory-based reporting, developed by the UPMC Health System, Pittsburgh, Pennsylvania, was evaluated to determine if it could be integrated into the conventional paper-based reporting system. We reviewed reports of 10 infectious diseases from 8 UPMC hospitals that reported to the Allegheny County Health Department in southwestern Pennsylvania during January 1-November 26, 2000. Electronic reports were received a median of 4 days earlier than conventional reports. The completeness of reporting was 74% (95% confidence interval [CI] 66% to 81%) for the electronic laboratory-based reporting and 65% (95% CI 57% to 73%) for the conventional paper-based reporting system (p>0.05). Most reports (88%) missed by electronic laboratory-based reporting were caused by using free text. Automatic reporting was more rapid and as complete as conventional reporting. Using standardized coding and minimizing free text usage will increase the completeness of electronic laboratory-based reporting. PMID- 12095437 TI - Temporal changes in prevalence of antimicrobial resistance in 23 US hospitals. AB - Antimicrobial resistance is increasing in nearly all health-care-associated pathogens. We examined changes in resistance prevalence during 1996-1999 in 23 hospitals by using two statistical methods. When the traditional chi-square test of pooled mean resistance prevalence was used, most organisms appear to have increased in prevalence. However, when a more conservative test that accounts for changes within individual hospitals was used, significant increases in prevalence of resistance were consistently observed only for oxacillin-resistant Staphylococcus aureus, ciprofloxacin-resistant Pseudomonas aeruginosa, and ciprofloxacin- or ofloxacin-resistant Escherichia coli. These increases were significant only in isolates from patients outside intensive-care units (ICU). The increases seen are of concern; differences in factors present outside ICUs, such as excessive quinolone use or inadequate infection-control practices, may explain the observed trends. PMID- 12095436 TI - Infection by Ralstonia species in cystic fibrosis patients: identification of R. pickettii and R. mannitolilytica by polymerase chain reaction. AB - The frequency of respiratory tract infections caused by Ralstonia species in persons with cystic fibrosis (CF) and the role of these species in CF pulmonary disease are not well documented. In part, this lack of documentation may be attributed to the difficulty in accurately identifying Ralstonia species; R. mannitolilytica and R. pickettii in particular may be misidentified as other closely related species, particularly those of the Burkholderia cepacia complex. We used polyphasic analyses to identify 42 Ralstonia isolates from sputum cultures from 38 CF patients. Several isolates that could not be identified to the species level may belong to novel Ralstonia species. We demonstrated chronic colonization by using genotyping of serial isolates recovered from the same patient. To facilitate identification of R. mannitolilytica and R. pickettii, we developed 16S ribosomal DNA-based polymerase chain reaction assays that allow sensitive and specific identification of these species. PMID- 12095438 TI - Monitoring antimicrobial use and resistance: comparison with a national benchmark on reducing vancomycin use and vancomycin-resistant enterococci. AB - To determine if local monitoring data on vancomycin use directed quality improvement and decreased vancomycin use or vancomycin-resistant enterococci (VRE), we analyzed data from 50 intensive-care units (ICUs) at 20 U.S. hospitals reporting data on antimicrobial-resistant organisms and antimicrobial agent use. We compared local data with national benchmark data (aggregated from all study hospitals). After data were adjusted for changes in prevalence of methicillin resistant Staphylococcus aureus, changes in specific prescriber practice at ICUs were associated with significant decreases in vancomycin use (mean decrease -48 defined daily doses per 1,000 patient days, p<0.001). These ICUs also reported significant decreases in VRE prevalence compared with those not using unit specific changes in practice (mean decrease of 7.5% compared with mean increase of 5.7%, p<0.001). In this study, practice changes focused towards specific ICUs were associated with decreases in ICU vancomycin use and VRE prevalence. PMID- 12095439 TI - Prevalence, distribution, and host range of Peste des petits ruminants virus, Turkey. AB - Peste des petits ruminants virus (PPRV, genus Morbillivirus), which causes a severe disease in sheep and goats, has only recently been officially declared to be present in Turkey. We carried out a study to determine the prevalence, distribution, and host range of PPRV in Turkey. A total of 1,607 animals, reared in 18 different locations, were monitored for the presence of antibodies to PPRV and the related virus of large ruminants, Rinderpest virus (RPV). Only two farms had animals that were free of antibody responses to either disease. Prevalence for PPRV infection varied (range 0.87%-82.6%) and was higher in sheep (29.2%) than in goats (20%). The overall antibody responses to PPRV and RPV were 22.4% and 6.28%, respectively. Two PPRVs of lineage 4, which comprises many other PPRVs whose origins are in the Middle East, the Arabian Peninsula, and southern Asia, were isolated from Turkish sheep. PMID- 12095440 TI - Antibiotic resistance of gram-negative bacteria in rivers, United States. AB - Bacteria with intrinsic resistance to antibiotics are found in nature. Such organisms may acquire additional resistance genes from bacteria introduced into soil or water, and the resident bacteria may be the reservoir or source of widespread resistant organisms found in many environments. We isolated antibiotic resistant bacteria in freshwater samples from 16 U.S. rivers at 22 sites and measured the prevalence of organisms resistant to beta-lactam and non-beta-lactam antibiotics. Over 40% of the bacteria resistant to more than one antibiotic had at least one plasmid. Ampicillin resistance genes, as well as other resistance traits, were identified in 70% of the plasmids. The most common resistant organisms belonged to the following genera: Acinetobacter, Alcaligenes, Citrobacter, Enterobacter, Pseudomonas, and Serratia. PMID- 12095441 TI - Bear Canyon virus: an arenavirus naturally associated with the California mouse (Peromyscus californicus). AB - Thirty-four rodents captured in southern California were studied to increase our knowledge of the arenaviruses indigenous to the western United States. An infectious arenavirus was isolated from 5 of 27 California mice but none of the 7 other rodents. Analyses of viral nucleocapsid protein gene sequence data indicated that the isolates from the California mice are strains of a novel Tacaribe serocomplex virus (proposed name "Bear Canyon") that is phylogenetically most closely related to Whitewater Arroyo and Tamiami viruses, the only other Tacaribe serocomplex viruses known to occur in North America. The discovery of Bear Canyon virus is the first unequivocal evidence that the virus family Arenaviridae is naturally associated with the rodent genus Peromyscus and that a Tacaribe serocomplex virus occurs in California. PMID- 12095442 TI - Entomologic and serologic evidence of zoonotic transmission of Babesia microti, eastern Switzerland. AB - We evaluated human risk for infection with Babesia microti at a site in eastern Switzerland where several B. microti-infected nymphal Ixodes ricinus ticks had been found. DNA from pooled nymphal ticks amplified by polymerase chain reaction was highly homologous to published B. microti sequences. More ticks carried babesial infection in the lower portion of the rectangular 0.7-ha grid than in the upper (11% vs. 0.8%). In addition, we measured seroprevalence of immunoglobulin (Ig) G antibodies against B. microti antigen in nearby residents. Serum from 1.5% of the 396 human residents of the region reacted to B. microti antigen (>1:64), as determined by indirect immunofluorescence assay (IgG). These observations constitute the first report demonstrating B. microti in a human biting vector, associated with evidence of human exposure to this agent in a European site. PMID- 12095443 TI - Rickettsialpox in North Carolina: a case report. AB - We report a case of rickettsialpox from North Carolina confirmed by serologic testing. To our knowledge, this case is the first to be reported from this region of the United States. Including rickettsialpox in the evaluation of patients with eschars or vesicular rashes is likely to extend the recognized geographic distribution of Rickettsia akari, the etiologic agent of this disease. PMID- 12095445 TI - Role of electronic data exchange in an international outbreak caused by Salmonella enterica serotype Typhimurium DT204b. AB - From July through September 2000, patients in five European countries were infected with a multidrug-resistant strain of Salmonella Typhimurium DT204b. Epidemiologic investigations were facilitated by the transmission of electronic images (Tagged Image Files) of pulsed-field gel electrophoresis profiles. This investigation highlights the importance of standardized protocols for molecular typing in international outbreaks of foodborne disease. PMID- 12095444 TI - Mycobacterium avium subsp. paratuberculosis infection in a patient with HIV, Germany. AB - Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of Johne disease in ruminants, has been incriminated as the cause of Crohn disease in humans. We report the first case of human infection with MAP in a patient with HIV; infection was confirmed by obtaining isolates from several different specimen types. PMID- 12095446 TI - Novel measles virus genotype, East Timor and Australia. AB - Measles outbreaks in 1999 in Queensland and Victoria, Australia, were caused by a novel strain of clade G virus (proposed name g3). Epidemiologic and molecular evidence supports independent circulation of this virus in Queensland, northern Australia, in addition to importation of the virus by East Timor refugees seeking safe haven in Australia. PMID- 12095447 TI - First outbreak of dengue hemorrhagic fever, Bangladesh. AB - During the first countrywide outbreak of dengue hemorrhagic fever in Bangladesh, we conducted surveillance for dengue at a hospital in Dhaka. Of 176 patients, primarily adults, found positive for dengue, 60.2% had dengue fever, 39.2% dengue hemorrhagic fever, and 0.6% dengue shock syndrome. The Dengue virus 3 serotype was detected in eight patients. PMID- 12095448 TI - Detection of West Nile virus in oral and cloacal swabs collected from bird carcasses. AB - We evaluated if postmortem cloacal and oral swabs could replace brain tissue as a specimen for West Nile virus (WNV) detection. WNV was detected in all three specimen types from 20 dead crows and jays with an average of >10(5) WNV PFU in each. These findings suggest that testing cloacal or oral swabs might be a low resource approach to detect WNV in dead birds. PMID- 12095449 TI - Smallpox research activities: U.S. interagency collaboration, 2001. PMID- 12095450 TI - High risk for tuberculosis in hospital physicians, Peru. PMID- 12095451 TI - First documented human Rickettsia aeschlimannii infection. PMID- 12095452 TI - Cost-effective screening for trichomoniasis. PMID- 12095454 TI - Hot-tub-associated mycobacterial infections in immunosuppressed persons. PMID- 12095455 TI - A method for comparing and combining cost-of-illness studies: an example from cardiovascular disease. AB - This paper describes a method for comparing and combining the results of various cost-of-illness (COI) studies. The method consists of seven steps: identify the study design; stratify according to the cost components; create concatenated cost components; adjust for inflation; adjust for population growth; compare cost estimates; and combine cost estimates. Based on this method, and using published data from 1986, 1993 and 1994, the cost of cardiovascular disease was estimated to be $20.1 billion in Canada in 2000, or $653 per person per year. One cost component, premature mortality, was found to have significantly decreased over time. The method described in this paper is sophisticated yet simple to use, and provides an efficient way to update, compare and combine cost estimates. By analyzing changes in cost components over time, it contributes to the projection methodology of cost information from multiple COI studies. The method greatly facilitates economic impact analyses to provide up-to-date information for healthy public policies. PMID- 12095456 TI - Cancer incidence in young adults in Canada: preliminary results of a cancer surveillance project. AB - Surveillance of cancer in young adults has been neglected, despite Sir Richard Doll's having emphasized its importance a decade ago. This report describes the patterns, time trends and regional variation in cancer incidence in Canada's young adults. In 1987 96, 97,469 cancers were diagnosed in Canadians aged 20 44, with almost two-thirds in females. Ten types of cancer accounted for 83% of diagnoses in women and 74% in men. The most common cancers in young women were breast, cervix, melanoma, thyroid and ovary, and in young men were testis, non- Hodgkin's lymphoma, melanoma, colorectal and lung. Although incidence rose only slightly for total cancer between 1969 and 1996, it increased dramatically for several specific types of cancer: lung (women), melanoma, testis, thyroid and non Hodgkin's lymphoma. Incidence declined for a few cancers (colorectal, lung (men), cervix and ovary). Lung cancer incidence was significantly lower than the Canadian average in Prairie women and non-significantly high in Quebec (both sexes), while the rate of melanoma was significantly low in Quebec (both sexes) and high in women in the Pacific region. PMID- 12095457 TI - Use of focus group methodology in the development of an Ontario farmers' sun safety survey. AB - Farmers are at higher risk for skin cancer; US studies indicate that they do not use adequate sun protection. Little data on Canadian farmers' sun exposure are available, and a literature review suggests a strong need to develop a comprehensive, easy to complete farmers' sun safety survey in order to identify sun safety issues in the farming community. A literature review contributed to the development of a draft farmers' sun safety survey. Preliminary testing of the survey with 207 Ontario farmers supported the usefulness of the questionnaire, but weaknesses remained in phrasing and missed concepts. To augment the questionnaire's development, focus groups were held with farmers in four Ontario communities to clarify the phrasing of survey questions concerning the amount of sun exposure, the use of sun protection practices, family/personal history of skin cancer, and skin cancer attitudes and knowledge. This paper reports on what was learned substantively from these focus groups. PMID- 12095458 TI - An integrated exploration into the social and environmental determinants of health: the Saskatchewan Population Health and Evaluation Research Unit (SPHERU). AB - The Saskatchewan Population Health and Evaluation Research Unit (SPHERU) is a new interdisciplinary research institute established by the Universities of Saskatchewan and Regina. SPHERU developed four of its research programs using a hierarchic model of health determining conditions and contexts. In descending order these programs include: Economic and Environmental Globalization, Governance and Health Community/Environment as a Health Determinant Multiple Roles, Gender and Health Determinants of Healthy Childhood Development A fifth program researching the determinants of health of indigenous peoples spans all four levels. Two research projects, one on power, control and health, and another on community capacity building approaches to human service programs, assist SPHERU in developing the theoretical linkages between its programs. This article describes SPHERU's research model and the Unit's approach to research and summarizes each of its current research programs and projects. PMID- 12095459 TI - Horse kicks, anthrax and the Poisson model for deaths. PMID- 12095460 TI - Peripheral and central sensitization in fibromyalgia: pathogenetic role. AB - Characteristic symptoms of fibromyalgia syndrome include widespread pain, fatigue, sleep abnormalities, and distress. Patients with fibromyalgia show psychophysical evidence of mechanical, thermal, and electrical hyperalgesia. Peripheral and central abnormalities of nociception have been described in fibromyalgia. Important nociceptor systems in the skin and muscles seem to undergo profound changes in patients with fibromyalgia through unknown mechanisms. They include sensitization of vanilloid receptor, acid-sensing ion channel receptors, and purino-receptors. Tissue mediators of inflammation and nerve growth factors can excite these receptors and cause extensive changes in pain sensitivity, but patients with fibromyalgia lack consistent evidence for inflammatory soft tissue abnormalities. Therefore, recent investigations have focused on central nervous system mechanisms of pain in fibromyalgia. PMID- 12095461 TI - Neuropharmacologic targets and agents in fibromyalgia. AB - Chronic widespread pain is a primary feature of fibromyalgia and is a symptom that is poorly managed in many patients. In addition, patients often experience fatigue, sleep disturbances, and anxiety. Its etiology is largely unknown. The successful clinical management of this syndrome relies on a multidisciplinary approach, employing pharmacologic and nonpharmacologic treatments. Clinical evidence on the symptoms and characteristics of fibromyalgia suggests a central mechanism behind the pathogenesis of this syndrome. It is likely that drugs with central actions will prove to be effective against a number of symptoms. This article aims to outline some of the potential spinal pharmacologic targets that may be used to treat this condition. PMID- 12095462 TI - Myofascial pain syndrome and its suggested role in the pathogenesis and treatment of fibromyalgia syndrome. AB - Myofascial pain syndrome is a chronic muscle pain disorder in one or more muscles or groups of muscles accompanied by local and referred pain, decreased range of motion, weakness, and often autonomic phenomena. Patients are readily recognized by their history of muscle pain and the presence of myofascial trigger points, which are specific areas of hyperirritability in a muscle that cause local and referred pain on palpation. Failure to recognize MPS often leads to over investigation, unnecessary medical intervention, and iatrogenic harm with serious cost implications. The purpose of this review is to present clinically relevant data regarding myofascial pain syndrome and to discuss its possible role in the pathophysiology and optimal treatment of fibromyalgia syndrome. PMID- 12095463 TI - Rheumatic mimics and selected triggers of fibromyalgia. AB - Fibromyalgia is a chronic pain syndrome of unknown etiology characterized by diffuse pain and tender points, which have been present for more than 3 months. Many patients with systemic illnesses can have diffuse pain similar to that found in fibromyalgia, including rheumatic diseases such as polymyalgia rheumatica, rheumatoid arthritis, idiopathic inflammatory myopathy, systemic lupus erythematosus, and joint hypermobility. Osteomalacia and thyroid disease are also in the differential diagnosis of diffuse pain and are imminently treatable. In addition, there has been interest throughout the past 10 years in infectious diseases including hepatitis C, Lyme disease, coxsackie B, HIV, and parvovirus infection, which may cause or trigger fibromyalgia. This paper provides a framework to use when identifying these diseases as part of the evaluation of a patient with chronic widespread musculoskeletal pain. PMID- 12095465 TI - The relationship of neck injury and post-traumatic headache. AB - Although there may be several causes of post-traumatic headache, neck injury is perhaps the most common. This paper primarily reviews the relationship of neck injury, whiplash, and post-traumatic headache. Mechanisms may include structural damage from acceleration or extension of the neck, development of myofascial pain and trigger points, interaction of the trigeminal nociceptive system with the upper cervical (occipital) nerves, and psychologic and emotional factors. Although some healing will occur, the outcome may depend on a number of human factors (awareness of an impending collision) and the fact that repaired tissue is different from normal, uninjured tissue. PMID- 12095464 TI - Neuroendocrine abnormalities in fibromyalgia. AB - Fibromyalgia is a disorder of unknown etiology characterized by chronic, widespread musculoskeletal pain and symptoms such as fatigue, poor sleep, gastrointestinal complaints, and psychologic problems that are similar to those experienced by patients with hormone deficiencies. This review summarizes the available data on the neuroendocrine function in fibromyalgia, including data on hormone secretion, circadian phase, and autonomic nervous system function. Studies suggest that there may be lower activity of a number of hypothalamic pituitary-peripheral gland axes and altered autonomic nervous system function in patients with fibromyalgia. These reductions in activity are mild to moderate and do not result from alterations in circadian rhythms. The reduced hormonal and autonomic responses appear to reflect an impairment in the hypothalamic or central nervous system response to stimuli rather than a primary defect at the level of the pituitary gland or the peripheral glands. A combination of multiple, mild impaired responses may lead to more profound physiologic and clinical consequences as compared with a defect in only one system, and could contribute to the symptoms of fibromyalgia. PMID- 12095466 TI - Cervicogenic headache: interventional, anesthetic, and ablative treatment. AB - Cervicogenic headache is becoming an accepted clinical syndrome in which headache pain is thought to originate from the cervical spine. Unfortunately, there are no diagnostic imaging techniques of the cervical spine and associated structures that can determine the exact source of pain. Therefore, diagnosis and treatment are based on the major accepted criteria of clinical presentation and the use of diagnostic nerve blocks to identify the source of the pain generator before considering further interventional or neuroablative treatment. This suggests that consistent reproducible anatomic and neurophysiologic pathways exist for the reproduction of typical clinical pain patterns and the ability of neuroblockade to consistently interrupt these pain pathways. This article describes the essential anatomy required to understand the use of diagnostic nerve blocks, and their predictive value in anticipating response to neuroablative and interventional therapy with a review of the major interventional, anesthetic, and ablative techniques for cervicogenic headache. PMID- 12095468 TI - The use of botulinum toxin in the treatment of headaches. AB - The use of botulinum toxin for movement disorders and cosmesis led to an accidental discovery of its beneficial effect on headaches. Extensive anecdotal evidence and several controlled trials suggest that intermittent and chronic migraines and chronic tension headaches may respond to this treatment. The effect of a single treatment, which is simple to administer, can last for 3 months. Botulinum toxin does not cause systemic or any other serious side effects. Prophylactic pharmacotherapy of migraine headaches is limited in its efficacy and has a potential for systemic side effects. This makes botulinum toxin a preferred treatment for many patients. The large controlled trials that are underway may lead to a wider acceptance of this treatment by neurologists and pain specialists. PMID- 12095467 TI - Inflammatory mechanisms in cervicogenic headache: an integrative view. AB - Cervicogenic headache is a relatively common and still controversial form of headache that originates from the neck structures. The pathophysiology probably results from various local pain-producing factors, such as intervertebral dysfunction, but the frequent coexistence of a history of head traumas still plays an important role. This report represents a series of pathophysiologic studies performed for patients with cervicogenic headache and the results achieved by a new pharmacologic treatment for the disease. PMID- 12095469 TI - Cervical myofascial pain and headache. AB - Myofascial pain is a common cause of regional chronic pain. Myofascial trigger points can refer pain to the head and face in the cervical region, thus contributing to cervicogenic headache. When identified properly, cervical myofascial pain is a treatable component of headache management. This article reviews current literature on the pathophysiology, diagnosis, and management of cervical myofascial pain. PMID- 12095471 TI - Nausea and Vomiting. AB - Nausea and vomiting are rather stereotyped symptoms. The challenge is that nausea and vomiting have many different causes and, in some patients, management may be rather complex. The clue is to determine the causal factor early. It helps to separate acute vomiting (<48 hours onset) from chronic vomiting. In acute vomiting, the causal factor or factors are most often evident. Symptomatic treatment with parenteral central-type antiemetics is the preferred treatment. Histamine-1 receptor antagonists, phenothiazines, butyrophenones, and corticosteroids are suitable drugs. For specific types of acute vomiting, for example, chemotherapy-related vomiting, the 5-HT3 receptor antagonists are costlier but effective drugs with minimal side effects. Sometimes, oral and parenteral administration of the above-mentioned drugs may be combined. The origin of chronic vomiting is often obscure, requires specialized investigation, and the causative factor may be uncorrectable. Symptomatic treatment requires a value judgement. If delayed gastric emptying is a contributing factor, prokinetic agents (metoclopramide, erythromycin, cisapride in special cases, if authorized) may prove useful. Otherwise, symptomatic treatment with central antiemetics is the only recourse. Some patients with unexplained vomiting present with psychological disturbances that act as magnifying or contributing factors and may be helped by psychotherapeutics. PMID- 12095470 TI - Headache and Chiari I malformation: clinical presentation, diagnosis, and controversies in management. AB - The Chiari I malformation is a cerebellar anomaly of uncertain origin, defined in part as tonsillar herniation of at least 3 to 5 mm below the foramen magnum; it is associated with crowding of the craniocervical junction. It is the most frequent of the Chiari malformations and is associated with headaches, syringohydromyelia, and hydrocephalus. The clinical manifestations are related to direct compression of neural tissues and abnormal cerebrospinal fluid dynamics. Common presentation is in adulthood, but there is increasing recognition in childhood. Incidental magnetic resonance imaging discovers Chiari I in one third of patients who do not have clinical symptoms. Headaches in the occipital suboccipital region or those that are of cough-type suggest symptomatic Chiari I malformation. Suboccipital craniectomy is performed for patients with Chiari I malformation who have neural compression syndromes of the brainstem and spinal cord, select headache types, and other uncommon conditions that are not the topic of this review. PMID- 12095472 TI - Functional Heartburn. AB - Functional heartburn is defined as the presence of a retrosternal burning sensation in the absence of pathologic gastroesophageal reflux. The underlying pathophysiology seems to be heterogeneous. In a subset of patients, esophageal hypersensitivity to physiologic esophageal acid exposure is involved, and this is likely to respond to intensive antireflux treatment. Antireflux surgery was not studied systematically in these patients and should be considered only in exceptional cases. In the remaining patients, non-acid reflux, esophageal mechanosensitivity, and psychological factors may be involved. Treatment options in these patients are limited owing to a lack of studies. Emerging therapies include selective serotonin reuptake inhibitors and relaxation therapy. PMID- 12095474 TI - Irritable Bowel Syndrome. AB - Because treatment of irritable bowel syndrome (IBS) patients can be frustrating to the clinician and patient as well, the physician should strive to gain the patient's confidence with a concise, appropriate work-up and by offering reassurance and education that IBS is a functional disorder without significant long-term health risks. First-line treatment should be aimed at treating the most bothersome symptom. Tricyclic antidepressants are superior to placebo in reducing abdominal pain scores, as well as improving global symptom severity. Loperamide is superior to placebo in managing IBS-associated diarrhea. Whereas fiber has a role in treating constipation, its value for IBS or, specifically, in the relief of abdominal pain or diarrhea associated with IBS is controversial. Although certain antispasmodics have demonstrated superiority over placebo in managing abdominal pain, none of these agents are available in the United States. Probiotic therapy using Lactobacillus plantarum has demonstrated superiority to placebo in improving pain, regulating bowel habits, and decreasing flatulence. As studied in a recent placebo-controlled prospective study, Chinese herbal medicines significantly improved bowel symptom scores and global symptom profile, and reduced IBS-related quality of life impairment. Some of the most promising emerging therapies in IBS revolve around targeted pharmacotherapeutic modulation of serotonin receptors (ie, 5-HT3 and 5-HT4 subtypes), which are involved in sensory and motor functions of the gut. Other investigational agents that are also being explored include cholecystokinin antagonists, alpha2-adrenergic agonists (eg, clonidine), serotonin reuptake inhibitors (eg, citalopram), and neurokinin antagonists. IBS is best understood through the biopsychosocial paradigm, and therefore, its effective management requires a comprehensive multidisciplinary approach based on patient education and reassurance, enhanced by diet recommendations and lifestyle modifications, and complemented by pharmacotherapy and psychosocial intervention in more severe cases. PMID- 12095473 TI - Aerophagia and Intestinal Gas. AB - Aerophagia refers to a rather rare disorder that may occur in both children and adults that features repetitive air swallowing and belching and that may result in abdominal distention. There are few, if any, controlled studies to guide therapy, which remains largely supportive but may include behavioral therapy and psychotherapy. Bloating, distention, and other gas-related symptoms are common in functional gastrointestinal disorders, including the irritable bowel syndrome; their pathophysiology remains, for the most part, poorly understood. Two separate phenomena need to be distinguished in these disorders: gas production and gas perception. Thus, whereas gas production, which relates most closely to flatus emissions, is probably within the normal range in most patients with irritable bowel syndrome, gas transport or transit through the gut may be impaired and may lead to the retention of gas within segments of the gut. Visceral hypersensitivity, a common phenomenon in all functional disorders, may exacerbate the sensation of distention and contribute to other "gas-related" symptoms. Few controlled studies have addressed any of these issues. Although, on an empiric basis, dietary therapy may be partially effective in some situations, there is at present no data to support the use of any form of pharmacologic, endoscopic, or surgical therapy for any of these symptoms. PMID- 12095475 TI - Slow Transit Constipation. AB - The diagnosis of slow transit functional constipation is based upon diagnostic testing of patients with idiopathic constipation who responded poorly to conservative measures such as fiber supplements, fluids, and stimulant laxatives. These tests include barium enema or colonoscopy, colonic transit of radio-opaque markers, anorectal manometry, and expulsion of a water-filled balloon. Plain abdominal films can identify megacolon, which can be further characterized by barium or gastrografin studies. Colonic transit of radio-opaque markers identifies patients with slow transit with stasis of markers in the proximal colon. However, anorectal function should be characterized to exclude outlet dysfunction, which may coexist with colonic inertia. Because slow colonic transit is defined by studies during which patients consume a high-fiber diet, fiber supplements are generally not effective, nor are osmotic laxatives that consist of unabsorbed sugars. Stimulant laxatives are considered first-line therapy, although studies often show a diminished colonic motor response to such agents. There is no evidence to suggest that chronic use of such laxatives is harmful if they are used two to three times per week. Polyethylene glycol with or without electrolytes may be useful in a minority of patients, often combined with misoprostol. I prefer to start with misoprostol 200 mg every other morning and increase to tolerance or efficacy. I see no advantage in prescribing misoprostol on a TID or QID basis or even daily because it increases cramping unnecessarily. This drug is not acceptable in young women who wish to become pregnant. An alternative may be colchicine, which is reported to be effective when given as 0.6 mg TID. Long-term efficacy has not been studied. Finally, biofeedback is a risk-free approach that has been reported as effective in approximately 60% of patients with slow transit constipation in the absence of outlet dysfunction. Although difficult to understand conceptually, it is worth attempting and certainly so in patients with associated pelvic floor dyssynergia. Subtotal colectomy with ileorectal anastomosis is often effective in those patients with colonic inertia, normal anorectal function, and lack of evidence of generalized intestinal dysmotility. However, morbidity is significant both early and late in the disease process and must be balanced against current disability. Ileostomy is preferred in the presence of anorectal dysfunction or with associated impairment of continence mechanisms. Similar considerations apply to the patient with disabling functional megacolon. An alternative approach is ileostomy with disconnection of the colon, which is more acceptable to some patients who may hope for future reconnection if recovery occurs. An additional alternative approach for patients with colonic inertia or megacolon who are not good surgical risks is tube cecostomy (or in children, use of the appendix as a conduit to the cecum). This permits either decompression (in megacolon) or antegrade enemas (in colonic inertia). Our surgeons are not enthusiastic about this approach, and I have little experience with it. In general, the use of partial resections of the colon should be discouraged, because marker studies do not define pathophysiology in patients with slow transit constipation. PMID- 12095476 TI - Biliary Dyskinesia. AB - Biliary dyskinesia is a motility disorder that affects the gallbladder and sphincter of Oddi. The motility disorder of the gallbladder is called gallbladder dyskinesia. Patients with this condition present with biliary-type pain, and investigations show no evidence of gallstones in the gallbladder. The diagnosis is made by performing a gallbladder ejection fraction, which is a radionuclide investigation. An abnormal gallbladder ejection fraction has a value less than 40%. Patients with an abnormal gallbladder ejection fraction should undergo cholecystectomy. This procedure has been shown to be effective in curing the symptoms in over 90% of patients. Motility disorder of the sphincter of Oddi is called sphincter of Oddi dysfunction. This disorder is categorized as two distinct types--biliary sphincter of Oddi dysfunction and pancreatic sphincter of Oddi dysfunction. Typically, patients with biliary sphincter of Oddi dysfunction present with biliary-type pain on average 4 to 5 years after having undergone cholecystectomy. Sphincter of Oddi manometry is essential in making a diagnosis of abnormal motility of the sphincter. On manometry, diagnosis of a sphincter of Oddi stenosis should lead to division of the sphincter. Sphincterotomy results in long-term relief of symptoms in more than 80% of patients. Pancreatic sphincter of Oddi dysfunction clinically presents with recurrent episodes of pancreatitis of unknown cause. Having ruled out all of the common causes of pancreatitis, sphincter of Oddi manometry of the pancreatic duct sphincter should be performed. When manometric stenosis is diagnosed, these patients should undergo division of both the biliary and pancreatic duct sphincter. This treatment results in relief of symptoms in more than 80% of patients. PMID- 12095477 TI - Serotonergic Agents in the Treatment of Functional Gastrointestinal Disorders. AB - Functional gastrointestinal disorders are exceptionally common, cause a great deal of suffering, and are extremely challenging to treat. Current therapies, usually targeted at possible abnormalities of gut contractility, are often disappointing and there is a real need for a new approach to treatment. Over the last decade, there has been an increase in our understanding of potential pathophysiologic processes involved in these conditions resulting in novel therapeutic agents based on this information. Serotonin receptor modulators are one such example of this approach, but their development has met with a number of difficulties mainly centered around issues of safety and trial design. It is becoming increasingly apparent that a drug universally effective in all forms of functional gastrointestinal disorders is unlikely to emerge unless some centrally acting compound can achieve this goal. Thus, the current approach of targeting specific subgroups is probably the best strategy. Many patients with functional gastrointestinal disorders do not even bother to consult the medical profession because of the lack of benefit from any currently available treatments. Thus, if a new effective drug is developed, even for a subgroup, demand is likely to be substantial, leading to an even greater burden on already stretched drug budgets. PMID- 12095478 TI - Pharmacologic Management of Gastroesophageal Reflux Disease. AB - The burden of gastroesophageal reflux disease (GERD) results from its widespread prevalence and the unfavorable impact of its symptoms on well-being and quality of life. Whereas abnormalities of the antireflux barrier (lower esophageal sphincter) are important in the pathophysiology of GERD, pharmacologic therapy for GERD is based on suppression of acid, which is responsible for the majority of the symptoms and for epithelial damage. Proton pump inhibitors (PPIs) are the agents of choice for achieving the goals of medical therapy in GERD, which include symptom relief, improvement in quality of life, and healing and prevention of mucosal injury. As a class, these drugs are extremely safe. The newest PPI, esomeprazole, brings a statistically significant increase in healing of mucosal injury and symptom relief in patients with erosive esophagitis, compared with omeprazole and lansoprazole. This article reviews the role of medical therapy in the short- and long-term management of symptomatic patients with or without erosive esophagitis, including extraesophageal presentations, GERD during pregnancy, and Barrett's esophagus. Management of refractory patients is addressed. PMID- 12095479 TI - Probiotics and Functional Foods in Gastrointestinal Disorders. AB - Probiotics are live microbial food supplements that benefit the host animal by improving intestinal microbial balance. When they are fed in yogurts, they can fall into the category of functional foods. Functional foods include these probiotics, prebiotics, and, to a certain extent, dietary fiber. Prebiotics are nondigestible food ingredients or supplements that alter the intestinal flora and stimulate the growth of healthy bacteria. Dietary fibers are part of plant foods that are nonstarch polysaccharides and are poorly digested or not digested by human enzymes. The physiologic process in which probiotics and functional foods affect the intestinal flora is through the balance of the intestinal microecology. This review looks at the four major components of intestinal microecology and describes the probiotics in use today and their clinical relevance. Although probiotics hold great promise and appear to be useful in some settings, more clinical study is needed to firmly establish the relevance of probiotic therapy. PMID- 12095481 TI - Molecular aspects of embryonic hemoglobin function. AB - In order to provide the appropriate level of oxygen transport to respiring tissues, we need to produce a molecular oxygen transporting system to supplement oxygen diffusion and solubility. This supplementation is provided by hemoglobin. The role of hemoglobin in providing oxygen transport from lung to tissues in the adult is well-documented and functional characteristics of the fetal hemoglobin, which provide placental oxygen exchange, are also well understood. However the characteristics of the three embryonic hemoglobins, which provide oxygen transport during the first three months of gestation, are not well recognized. This review seeks to describe the state of our understanding of the temporal control of the expression of these proteins and the oxygen binding characteristics of the individual protein molecules. The modulation of the oxygen binding properties of these proteins, by the various allosteric effectors, is described and the structural origins of these characteristics are probed. PMID- 12095482 TI - Interactions of cytokines, growth factors, and the extracellular matrix in the cellular biology of uterine leiomyomata. AB - OBJECTIVE: To review the available information regarding the role of cytokines, growth factors, and the extracellular matrix in the pathophysiology of uterine leiomyomata and to integrate this information in a suggested model of disease at the cellular level. DESIGN: A thorough literature and MEDLINE search was conducted to identify the relevant studies in the English literature published between January, 1966 and October, 2001. A model of disease at the cellular level was developed using the most likely cytokines to be involved in the pathogenesis of leiomyomata as determined by our assessment of the available literature. RESULT(S): A number of cytokines and growth factors, including transforming growth factor-beta (TGF-beta), epidermal growth factor, monocyte chemotactic protein-1, insulin-like growth factors 1 and 2, prolactin, parathyroid-hormone related peptide, basic fibroblast growth factor, platelet-derived growth factor, interleukin-8, and endothelin, have been investigated in myometrium and leiomyoma. Among these cytokines, TGF-beta appears to be the only growth factor that has been shown to be overexpressed in leiomyoma vs. myometrium, be hormonally-regulated both in vivo and in vitro, and be both mitogenic and fibrogenic in these tissues. In addition to the cytokines, extracellular matrix components such as collagen, fibronectin, proteoglycans, matrix metalloproteinases, and tissue inhibitors of metalloproteinases seem to play pivotal roles in the pathogenesis of leiomyomata. CONCLUSION(S): We believe that, given the extent and depth of the current research on the cellular biology of leiomyomata, the cellular mechanisms responsible in the pathogenesis of leiomyomata will be identified clearly within the foreseeable future. This will enable researchers to develop therapy directed against the molecules and mechanisms at the cellular level. PMID- 12095483 TI - Assisted reproductive technology in developing countries: why should we care? PMID- 12095484 TI - Prospective, randomized, blinded evaluation of donor semen quality provided by seven commercial sperm banks. AB - OBJECTIVE: To evaluate variability in donor semen quality between seven commercial donor sperm banks, within sperm banks, and between intracervical insemination and intrauterine insemination. DESIGN: Prospective, randomized, blind evaluation of commercially available donor semen samples. SETTING: An academic andrology laboratory. PATIENT(S): Seventy-five cryopreserved donor semen samples were evaluated. INTERVENTION(S): Samples were coded, then blindly evaluated for semen quality. MAIN OUTCOME MEASURE(S): Standard semen quality parameters, including concentration, motility parameters, World Health Organization criteria morphology, and strict criteria morphology. RESULT(S): Significant differences were observed between donor semen banks for most semen quality parameters analyzed in intracervical insemination samples. In general, the greatest variability observed between banks was in percentage progressive sperm motility (range, 8.8 +/- 5.8 to 42.4 +/- 5.5) and normal sperm morphology (strict criteria; range, 10.1 +/- 3.3 to 26.6 +/- 4.7). Coefficients of variation within sperm banks were generally high. CONCLUSION(S): These data demonstrate the variability of donor semen quality provided by commercial sperm banks, both between banks and within a given bank. No relationship was observed between the size or type of sperm bank and the degree of variability. The data demonstrate the lack of uniformity in the criteria used to screen potential semen donors and emphasize the need for more stringent screening criteria and strict quality control in processing samples. PMID- 12095485 TI - Use of the total motile sperm count to predict total fertilization failure in in vitro fertilization. AB - OBJECTIVE: To evaluate the capacity of baseline characteristics and total motile sperm count (TMC) to predict total fertilization failure (TFF) in patients undergoing IVF. DESIGN: Retrospective cohort study. SETTING: University hospital. PATIENT(S): Eight hundred ninety-two couples with a total of 1,569 consecutive IVF cycles. INTERVENTION(S): Prewash and postwash TMC during fertility workup and at the time of ovum pickup (OPU). MAIN OUTCOME MEASURE(S): Analysis of logistic regression and the receiver operating characteristic curve were used to determine which variables could be used to predict TFF. RESULT(S): The area under the curve (AUC) for prewash TMC during fertility workup was 0.72, similar to a combination of pre- and postwash TMC. At the time of OPU, both pre- and postwash TMC had an AUC of 0.73. A model based on selected baseline characteristics (male age, number of IVF cycles, indication for IVF, and prewash TMC during fertility workup) had an AUC of 0.75. A model at the time of OPU, including the number of oocytes, had an AUC of 0.80. CONCLUSION(S): The use of both models, one before start of the IVF cycle and one at the time of OPU, allows an accurate prediction of the chance of TFF and is useful in counseling patients on whether to opt for IVF or ICSI. PMID- 12095486 TI - Success of intrauterine insemination in women aged 40-42 years. AB - OBJECTIVE: To determine how advancing female age decreases successful outcomes of intrauterine insemination (IUI) alone or combined with ovarian stimulation. DESIGN: Retrospective review. SETTING: Academic fertility center. PATIENT(S): Infertile men and women. INTERVENTION(S): Intrauterine insemination alone or combined with ovarian stimulation. MAIN OUTCOME MEASURE(S): Pregnancy rates, miscarriage rates, and live birth rates per insemination cycle according to female age. RESULT(S): The 1,117 cycles of IUI resulted in 217 pregnancies, for an overall pregnancy rate for all female ages of 19.4% and a live birth rate of 12.9% per cycle inseminated. The overall live birth rate per insemination declined with advancing maternal age. CONCLUSION(S): Advancing female age decreases successful outcomes with IUI. The live birth rate with IUI for women 40 42 years old (n = 82) was 9.8% per insemination and may demonstrate that IUI is an appropriate treatment for this age group of women. PMID- 12095487 TI - Comparison of stimulation with clomiphene citrate in combination with recombinant follicle-stimulating hormone and recombinant luteinizing hormone to stimulation with a gonadotropin-releasing hormone agonist protocol: a prospective, randomized study. AB - OBJECTIVE: To compare IVF-ET outcome with a new stimulation protocol using clomiphene citrate (CC) with recombinant FSH and LH to stimulation with the standard long GnRH-a protocol. DESIGN: Prospective randomized study. SETTING: Outpatient infertility clinic in Vienna, Austria. PATIENT(S): Two hundred ninety four infertile women undergoing IVF-ET; 154 IVF cycles stimulated with CC + recombinant FSH + recombinant LH (group A) and 140 cycles with long GnRH-a suppression + recombinant FSH (group B). INTERVENTION(S): Controlled ovarian hyperstimulation, egg retrieval, and ET. MAIN OUTCOME MEASURE(S): Cycle parameters (number of oocytes, fertilization, number of embryos) and outcome (pregnancy rate, cancellation rate, ovarian hyperstimulation syndrome [OHSS]). RESULT(S): Pregnancy rate per ET was 42.9% (implantation rate, 21.3%) in group A and 36.6% (17.4%) in group B. Cancellation rates were similar. The OHSS occurred in four cases (3%) in group A and 12 cases (10%) in group B. CONCLUSION(S): Stimulation with CC + recombinant FSH + recombinant LH leads to comparable pregnancy rates vs. the long protocol. With this new stimulation, less gonadotropins are used and there is less need for monitoring (lower cost for patient and clinic). The risk of OHSS is reduced as well. Therefore, this protocol should be regarded as the first-line treatment. PMID- 12095488 TI - Factors influencing the cumulative conception rate and discontinuation of in vitro fertilization treatment for infertility. AB - OBJECTIVE: To examine the cumulative conception rate and live birth rate in women undergoing IVF and to assess the influence of prognostic factors on cumulative conception rate and discontinuation of treatment. DESIGN: Retrospective analysis of data from couples undergoing IVF. SETTING: Assisted conception unit of a university hospital. PATIENT(S): Two thousand fifty-six patients undergoing 2708 cycles of IVF from April 1992 to March 1999. MAIN OUTCOME MEASURE(S): Cumulative conception rate by age, number of oocytes retrieved, and embryos transferred, and the influence of these factors on dropout rates. RESULT(S): The cumulative conception rate and cumulative live birth rate after four attempts were 75% and 66%, respectively. The cumulative conception rate differed significantly between women 35 years of age or younger and those older than 35 years who had five or more oocytes retrieved (83% vs. 63%). When fewer than five oocytes were retrieved in women 35 years of age or younger, the cumulative conception rate decreased to 33%. Overall, 36% of patients continued treatment after the first attempt; these patients were more likely to have more than five oocytes retrieved and more than two embryos available for transfer. CONCLUSIONS: The cumulative conception rate was greater when the female partner was 35 years of age or younger and had more than five oocytes retrieved and more than two embryos were available for transfer. These factors influenced dropout rates. PMID- 12095489 TI - Elevated serum progesterone-to-estradiol ratio during gonadotropin stimulation for intrauterine insemination or in vitro fertilization is not associated with diminished ovarian reserve. AB - OBJECTIVE: To determine whether an elevated serum P:E2 ratio on the day of hCG administration during gonadotropin stimulation for intrauterine insemination (IUI) or IVF is associated with diminished ovarian reserve. DESIGN: Retrospective chart review. SETTING: Tertiary fertility center. PATIENT(S): Two hundred eighty six women undergoing a fertility evaluation. INTERVENTION(S): Clomiphene citrate challenge test (CCCT). MAIN OUTCOME MEASURE(S): E2 and P concentrations on the day of hCG administration. RESULT(S): For the IUI group (n = 98), 74 women had a normal CCCT, while 24 had an abnormal CCCT. For women undergoing IVF (n = 188), 171 had a normal CCCT and 17 had an abnormal CCCT. For women undergoing IUI with a normal CCCT, 33 (44%) had a [(P/E2) x 1000] ratio <1 on the day of hCG, while 12 (50%) (abnormal CCCT) had a [(P/E2) x 1000] ratio <1. Similarly, for women doing IVF with a normal CCCT, 96 (56%) had a [(P/E2) x 1000] ratio <1 on the day of hCG, and 9 (44%) (abnormal CCCT) had a [(P/E2) x 1000] ratio <1 on the day of hCG. The frequencies were similar for all comparable groups. For both groups, there was no association between serum (P/E2) x 1000 on the day of hCG administration and day 3 or 10 FSH during the CCCT. For women with a normal CCCT, there were no differences in delivery rates based on the (P/E2) x 1000 ratio. CONCLUSION(S): In women stimulated with hMG for IUI or IVF, the serum P:E2 ratio on the day of hCG administration is not associated with diminished ovarian reserve, as demonstrated by the CCCT, or pregnancy outcome. PMID- 12095490 TI - Self-selected women with polycystic ovary syndrome are reproductively and metabolically abnormal and undertreated. AB - OBJECTIVE: To determine whether self-selected women with polycystic ovary syndrome (PCOS) are abnormal compared with a control population. DESIGN: Case control. SETTING: Support group meeting organized and initiated by patients. PATIENT(S): Forty-five self-selected women with PCOS and 80 control women. INTERVENTION(S): Self-selected women with PCOS at a peer support conference completed a questionnaire, had a brief physical, and gave a fasting blood sample. MAIN OUTCOME MEASURE(S): Historical, biometric, and assay results. RESULT(S): Sixty percent of the women attending the conference participated in the study. Most had been diagnosed with PCOS on the basis of ovarian morphology (35%). They were more likely to be nulliparous and have a history of oligomenorrhea (96%). They were hyperandrogenemic (significantly elevated testosterone and DHEAS levels) compared with control women. Self-selected women with PCOS displayed multiple metabolic abnormalities compared with control women, including elevations in blood pressure, waist-hip ratio, fasting insulin, fasting total cholesterol, and fasting low-density lipoprotein cholesterol levels, as well as a significant decrease in fasting glucose-insulin ratio and high-density lipoprotein cholesterol levels. CONCLUSION(S): Self-selected women with PCOS have reproductive and metabolic abnormalities. The majority of these women received inadequate treatment despite having risk factors for endometrial cancer, diabetes, and/or heart disease. Our study also suggests that women attending or participating in a PCOS support group are willing and likely to participate in clinical studies. PMID- 12095491 TI - Hereditary thrombophilias are not associated with a decreased live birth rate in women with recurrent miscarriage. AB - OBJECTIVE: To assesses the live birth rate without treatment in women with hereditary thrombophilia who have recurrent miscarriage and women without thrombophilia who have recurrent miscarriage. DESIGN: Prospective observational study. SETTING: Tertiary referral unit in university hospital. PATIENT(S): One hundred twenty women with thrombophilia and 65 women without thrombophilia. MAIN OUTCOME MEASURE(S): Number of live births or repeated miscarriages. RESULTS: Of the 185 patients, 44 with thrombophilia and 26 without thrombophilia have conceived. Nineteen of the 44 pregnancies (43.2%) in thrombophilia patients have terminated in live births, compared with 8 of 26 pregnancies (30.8%) in patients without thrombophilia. This difference is not statistically significant. CONCLUSIONS: Hereditary thrombophilia did not seem to affect the live birth rate in women with recurrent miscarriage. PMID- 12095492 TI - Bone metabolism in postmenopausal women who were treated with a gonadotropin releasing hormone agonist and tibolone. AB - OBJECTIVE: To study the bone metabolism in postmenopausal women who have been treated with gonadotropin-releasing hormone agonist (GnRH-a) and tibolone. DESIGN: Prospective, open, controlled clinical trial. SETTING: Department of Gynecology and Obstetrics, University of Catanzaro, Catanzaro, Italy. PATIENT(S): One hundred twenty perimenopausal women with symptomatic uterine leiomyomas (groups A and B), and 40 healthy control women who underwent a normal spontaneous menopause (group C). INTERVENTION(S): Treatment for 12 months with leuprolide acetate plus tibolone (group A) or hysterectomy with bilateral oophorectomy (group B). MAIN OUTCOME MEASURE(S): Lumbar spine bone mineral density (BMD) and bone turnover markers at entry into the study, after medical treatment (only group A), and 12 months after discontinuation medical treatment (group A) or after surgery (group B). The same parameters were noted in healthy women before and 12 months after menopause (retrospective control group, group C). RESULT(S): At the women's entry into the study, no significant difference in BMD and bone turnover markers was detected between groups A and B. In group A, no significant variation in BMD or bone turnover markers was observed 12 months after medical treatment in comparison with baseline. At 12 months after discontinuation of treatment (in women who had achieved menopause) and after surgery, we observed a statistically significant decrease in BMD and in bone turnover markers in both groups in comparison with baseline. At 12 months after they became menopausal, we also observed a statistically significant reduction in BMD and in bone turnover markers in control group C. At the same 12-month follow-up visit, a statistically significant difference in BMD and in bone turnover markers was detected when comparing groups A and B with group C. CONCLUSION(S): Women previously treated with GnRH-a and tibolone similar to women who are menopausal as a result of surgery, have higher bone loss after menopause. PMID- 12095493 TI - Marked elevation of macrophage migration inhibitory factor in the peritoneal fluid of women with endometriosis. AB - OBJECTIVE: To evaluate the presence of macrophage migration inhibitory factor (MIF) in the peritoneal fluid of normal fertile women and patients with endometriosis and its growth-promoting activity toward human endothelial cells. DESIGN: Retrospective study using ELISA to measure peritoneal fluid MIF, and [3H] thymidine incorporation into the DNA of human endothelial cells to assess its mitogenic activity. SETTING: Gynecology clinic and human reproduction research laboratory. PATIENT(S): Thirty-six healthy women and 57 women with endometriosis. INTERVENTION(S): Peritoneal fluid samples were obtained at laparoscopy. MAIN OUTCOME MEASURE(S): Macrophage migration inhibitory factor concentrations in the peritoneal fluid samples and [3H]-thymidine incorporation into the DNA of human microvascular endothelial cells to assess proliferation. RESULT(S): This study demonstrated the presence of MIF in the peritoneal fluid and a 238% increase of MIF levels in women with endometriosis as compared with healthy women. Both fertile and infertile women with endometriosis had significantly higher MIF concentrations than did fertile women with normal gynecological status, but the difference was more significant in infertile endometriosis patients. Anti-MIF antibody significantly inhibited proliferation of human microvascular endothelial cells in response to peritoneal fluids from healthy women and women with endometriosis stages I-II and III-IV, as assessed by [3H]-thymidine incorporation. CONCLUSION(S): This study revealed the presence of MIF in the peritoneal fluid and its increased levels in endometriosis and suggests that MIF may be involved in endometriosis-associated infertility and angiogenesis. PMID- 12095494 TI - Follicular development, ovulation, and corpus luteum formation in cryopreserved human ovarian tissue after xenotransplantation. AB - OBJECTIVE: To assess the competency of human frozen/thawed ovarian follicles matured in xenografts to form functioning corpora luteae after human chorionic gonadotropin (hCG) administration. DESIGN: Prospective controlled animal study. SETTING: University research laboratory. PATIENT(S): Three women (19, 28, and 36 years) who underwent oophorectomy. ANIMAL(S): Nineteen female severe combined immunodeficient (SCID) mice. INTERVENTION(S): Cryopreserved human ovarian tissues were grafted into the s.c. space of bilaterally oophorectomized SCID mice. All the animals were stimulated with pregnant mare's serum gonadotropin (PMSG) for 4 weeks starting from 16 weeks after transplantation. Twelve animals were injected with hCG at the end of gonadotropin stimulation. MAIN OUTCOME MEASURE(S): [1] The rate of grafts with growing follicles, with antral follicles, and/or with corpora luteae. [2] The histologic assessment of follicles and corpora luteae. [3] The serum progesterone and estradiol level in animals with corpus luteum in the grafts. RESULT(S): [1] The rate of grafts with growing follicles and with corpora luteae was 33% to 100%, and 28% to 50%, respectively. [2] Corpora luteae in xenografts were all morphologically normal. [3] The progesterone levels were all above 3.0 ng/mL. CONCLUSION(S): This study showed that the cryopreserved human ovarian follicles can be matured to a stage at which they can form functioning corpora luteae in the host animal. PMID- 12095495 TI - Influence of low-dose oral contraception on peripheral blood lymphocyte subsets at particular phases of the hormonal cycle. AB - OBJECTIVE: To investigate the effects of low-dose oral hormonal contraception on the immune system during certain phases of the hormonal cycle. DESIGN: Prospective, nonrandomized, controlled study. SETTING: Academic research setting. PATIENT(S): Women with regular menstrual cycle using hormonal oral contraception (OC; Cileste, 250 microg of norgestimat and 35 microg of ethinylestradiol, or Marvelon, 150 microg of desogestrel and 30 microg of ethinylestradiol) and women not using hormonal or other forms of contraception. INTERVENTION(S): Peripheral blood lymphocyte subsets were determined by flow cytometry on the first day of menstruation (day 1), in the follicular phase (day 8), midcycle (day 15), and in the luteal phase (day 22). MAIN OUTCOME MEASURE(S): Levels of lymphocyte subpopulations. RESULT(S): Women using OC had significantly higher levels of CD3+ CD8+ cells throughout their pill cycle compared to controls. Furthermore, women taking Cileste had lower levels of natural killer (NK) cells during their cycle and also women taking Marvelon but only from days 8-15. Within the pill cycle of Cileste we observed an increase in CD20+ and CD20+ CD5+ cells from days 1-8. CONCLUSION(S): Cytotoxic lymphocytes, which are responsible for first-line immune defense, and B cells, which are involved in autoimmune disorders, are affected by OC. PMID- 12095496 TI - Mean versus individual hormonal profiles in the menstrual cycle. AB - OBJECTIVE: To evaluate hormonal profiles of normal menstrual cycles. DESIGN: Prospective, descriptive study of a case series. SETTING: University-based natural family planning center. PATIENT(S): Twenty-five natural family planning users for three or more cycles (n = 78). These women were healthy, contraception free, parous, with regular ovulatory cycles. INTERVENTION(S): Immunoassays for estrone glucuronide, LH, and pregnanediol glucuronide were done in daily timed and measured samples of early morning urine. MAIN OUTCOME MEASURE(S): Estrone glucuronide, LH, and pregnanediol glucuronide levels were measured during the menstrual cycle. RESULT(S): All cycles showed an ovulatory pattern configuring classic hormonal mean curves. Most (77%) differed from the mean curve pattern. All had estrone glucuronide peaks, LH peaks, and pregnanediol glucuronide increases. Estrone glucuronide and LH peaks were not always clear; some lasted more than 1 day (long peak: estrone glucuronide 19%, LH 9%) or fluctuated (double peak: estrone glucuronide 4%, LH 6%; small LH peak: 19%). There were also prepeak estrone glucuronide surges, and pre- and postpeak LH surges. Pregnanediol glucuronide increased more clearly (6% fluctuated 1 day). Some women had repeated cycles with long estrone glucuronide peaks (16%) and fluctuations in LH surge (44%). CONCLUSION(S): Normal menstrual cycle hormonal profiles generally differ from mean curves, which are usually considered standard. PMID- 12095497 TI - Menstrual symptometrics: a simple computer-aided method to quantify menstrual cycle disorders. AB - OBJECTIVE: To validate a menstrual symptometrics device that can quantify menstrual blood loss, dysmenorrhea, and the premenstrual syndrome against traditional methods of collecting data on symptoms. DESIGN: Validation study. SETTING: Academic research clinic for menstrual cycle disorders. PARTICIPANT(S): Women 18-50 years of age who presented with menstrual cycle disorders. Controls were recruited from lists of patients requesting sterilization and from hospital staff. INTERVENTION(S): Participants were asked to complete the menstrual symptometrics device and to record pain, blood loss, and premenstrual symptoms by using traditional methods (paper-based scales and the alkaline hematin method) for two cycles. MAIN OUTCOME MEASURE(S): Agreement between traditional methods of quantifying menstrual cycle disorders and data obtained from the menstrual symptometrics device, and acceptability of the latter technique to patients. RESULT(S): A high level of agreement was observed between the traditional methods and the menstrual symptometrics device in quantifying and diagnosing menorrhagia, dysmenorrhea, and the premenstrual syndrome. Most patients preferred the menstrual symptometrics device as a data collection tool. CONCLUSION(S): The menstrual symptometrics device is a rapid and accurate method of quantifying blood loss, pain, and premenstrual symptoms. It has a high level of patient acceptability and can provide instant pictorial feedback on symptoms for patients and clinicians. PMID- 12095498 TI - Effects of initiation day of clomiphene citrate on the endometrium of women with regular menstrual cycles. AB - OBJECTIVE: To investigate the effects of initiation time of clomiphene citrate (CC) on the endometrium of women with regular menstrual cycles. DESIGN: Randomized, double-blind, cross-over study. SETTING: Department of Obstetrics and Gynecology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand. PATIENT(S): Thirty-three healthy female volunteers with regular menstrual cycles. INTERVENTION(S): The volunteers were randomized to receive either 100 mg of CC on days 1-5 and placebo on days 5-9 (study group) or placebo on days 1-5 and CC on days 5-9 (control group). After a wash-out period of 1 month of CC treatment, the medication was switched in each group. Ultrasonography was performed daily after day 10 of the cycle to detect ovulation. Ultrasonography for endometrial appearance and thickness, endometrial sampling, and blood samples obtained for determination of E(2) and P levels were performed 7 days after ovulation in both groups. MAIN OUTCOME MEASURE(S): Morphometric analysis, histologic dating, and ultrasonographic appearance and thickness of the endometrium. RESULT(S): Morphometric parameters, histologic dating, and ultrasonographic appearance and thickness of the endometrium were similar in both groups. CONCLUSION(S): Starting CC on either day 1 or day 5 of the menstrual cycle did not have any differential effects on the endometrium of women with regular menstrual cycles, particularly regarding the morphometric analysis, histologic dating, or ultrasonographic appearance. PMID- 12095499 TI - Effects of follicle-stimulating hormone (FSH) and human chorionic gonadotropin in individuals with an inactivating mutation of the FSH receptor. AB - OBJECTIVE: To study the gonadal steroid responses to FSH and hCG in individuals with the inherited Finnish-type inactivating Ala189Val mutation of the FSH receptor gene. DESIGN: Prospective clinical and descriptive study. SETTING: University hospital. PATIENT(S): Two women and one man homozygous for the Ala189Val mutation of the FSH receptor gene, and ovarian biopsies from four affected and four healthy women, and four normal fetuses. INTERVENTION(S): Individuals were treated with increasing doses of recombinant FSH (300 IU/day start, 900 IU/day final) and/or a single dose of hCG (5000 IU). Ovarian biopsies were used in immunohistochemical analyses for detection of aromatase cytochrome P450 and transcription factor GATA-4. In situ 3'-end labeling analyses were used for detection of apoptosis. MAIN OUTCOME MEASURE(S): Measurements of serum concentrations of follicle-stimulating hormone, leuteinizing hormone, inhibin A and B, estradiol, testosterone (T), androstenedione, and prolactin, immunostaining for ovarian aromatase, GATA-4, and apoptosis. RESULT(S): Administration of FSH had no effect on production of the steroids. Similarly, human chorionic gonadotropin (hCG) treatment, alone or after FSH administration, failed to raise serum steroid concentrations. Ovarian apoptosis was absent, and the expression of transcription factor GATA-4 and aromatase was negligible in the ovarian biopsies from Ala189Val homozygous individuals. CONCLUSION(S): The Ala189Val mutation of the FSH receptor gene results in a complete block of FSH action in vivo. Furthermore, the failure of hCG to increase both ovarian estradiol and testosterone secretion emphasizes the possible contribution of FSH in regulating ovarian androgen synthesis, and supports the concept that both gonadotropins are necessary for appropriate ovarian steroidogenesis in humans. PMID- 12095500 TI - Insights from gene arrays on the development and growth regulation of uterine leiomyomata. AB - OBJECTIVE: To use microarray analysis as an unbiased approach to identify genes involved in the induction and growth of uterine leiomyomata. DESIGN: Screen by arrays for up to 12,000 genes in leiomyoma (L) and control myometrium (M) from nine patients. SETTING: University research laboratories. PATIENT(S): Nine patients in the follicular and luteal phases of the menstrual cycle. INTERVENTION(S): mRNA from L and M was converted to biotin-labeled cRNA and hybridized to cDNA oligonucleotide sequences on the arrays. MAIN OUTCOME MEASURE(S): Greater than two-fold change in gene expression between leiomyoma and matched myometrium. RESULT(S): Prominent among the 67 genes overexpressed in L relative to M were dlk or Pref-1, doublecortin, JM27, ionotropic glutamate receptor subunit 2, apolipoprotein E3, IGF2, semaphorin F, myelin proteolipid protein, MEST, frizzled, CRABP II, stromelysin-3, and TGFbeta3. The genes dlk, IGF2, and MEST are paternally expressed imprinted genes, and the others are involved in tissue differentiation and growth. Prominent among the 78 genes down regulated in L relative to M were alcohol dehydrogenases 1alpha-gamma, tryptase, dermatopontin, thrombospondin, coxsackievirus receptor, nur77, and c-kit. CONCLUSION(S): Arrays offer large-scale screening of mRNA expression, which will help us differentiate between the genes and metabolic pathways necessary for leiomyoma growth and those regulating myometrial contractions. PMID- 12095501 TI - Medroxyprogesterone acetate does not antagonize estrogen-induced increases in endothelium-dependent vasodilation: potential clinical implications. AB - OBJECTIVE: Determine the effect of combined 17beta-estradiol benzoate (E2) and medroxyprogesterone acetate (MPA) administration on endothelium-dependent vasorelaxation to acetylcholine (ACh). DESIGN: Prospective, ex vivo study. SETTING: Academic research laboratory. ANIMAL(S): Mature female rats. INTERVENTION(S): Ovariectomized rats received one of the following interventions daily for 3 days: [1] corn oil via IM injection, [2] E2 (20 microg/kg IM), or [3] E2 (20 microg/kg IM) and MPA (10 mg/kg IM). MAIN OUTCOME MEASURE(S): Basal release of nitric oxide (NO) and endothelium-dependent and endothelium independent vasodilation from thoracic aortas obtained from each group. RESULT(S): Estradiol treatment potentiated the endothelium-dependent relaxation to ACh when compared with the control group. Administration of MPA with E2 did not antagonize the beneficial effect of E2 on endothelium-dependent relaxation. Estradiol treatment alone or in combination with MPA did not affect endothelium independent vasodilation as compared with the case of the control group. Administration of E2 resulted in increased basal NO release (assessed indirectly by measuring the constrictor response to N(G)-nitro-L-arginine [methyl ester (L NAME)]) when compared with the case of the control group, and the addition of MPA to E2 did not alter the effect of E2 on basal NO release. CONCLUSION(S): Estradiol potentiates endothelium-dependent relaxant responses and increases basal endothelial NO release. Medroxyprogesterone acetate does not antagonize these effects of E2. PMID- 12095502 TI - Phorbol myristate acetate induces ruffling of the acrosome of human sperm. AB - OBJECTIVE: To determine the effect of phorbol myristate acetate (PMA) on human acrosome morphology and the acrosome reaction. DESIGN: Controlled experiments on sperm and unfertilized oocytes from volunteers. SETTING: Academic research and teaching tertiary hospital. PATIENT(S): Sperm samples were from normospermic men and unfertilized oocytes from IVF patients. MAIN OUTCOME MEASURE(S): Acrosome morphology was assessed by using transmission and scanning electron microscopy. The acrosome reaction was assessed by using fluorescein-labeled Pisum sativum agglutinin. RESULT(S): PMA induced acrosome ruffling, indicated by a marked wavy appearance. A significant correlation was found between PMA-induced ruffling and PMA enhancement of the zona pellucida-induced acrosome reaction. Protein kinase C inhibitors bisindolylmalemide I and sangivamycin had no effect on PMA-induced acrosomal ruffling, but actin polymerization inhibitors cytochalasin B and cytochalasin D significantly decreased PMA-induced acrosomal ruffling. In contrast, bisindolylmalemide I, sangivamycin, cytochalasin B, and cytochalasin D significantly decreased both the zona pellucida-induced acrosome reaction and the PMA enhancement of the zona pellucida-induced acrosome reaction. CONCLUSION(S): PMA-induced acrosomal ruffling involves actin polymerization, possibly independent of conventional protein kinase C. Acrosomal ruffling is involved in the PMA augmentation of the zona pellucida-induced acrosome reaction. PMID- 12095503 TI - Apoptosis and proliferation of human peritoneal fibroblasts in response to hypoxia. AB - OBJECTIVE: To determine the effect of hypoxia on apoptosis and proliferation of fibroblasts cultured from normal peritoneum or adhesions. DESIGN: Prospective experimental study. SETTING: University medical center. PATIENT(S): Primary cultures of fibroblasts established from peritoneal and adhesion tissues of the same patients. INTERVENTION(S): Hypoxia treatment of the primary cultured fibroblasts. MAIN OUTCOME MEASURE(S): Apoptosis was assessed by the TUNEL assay and by the BCL-2-BAX expression ratio, as determined using multiplex reverse transcription polymerase chain reaction. The proliferation rate of these cells was determined by measuring the proliferative fraction using the flow cytometry in adhesion and peritoneal fibroblasts under normal and hypoxic conditions. RESULT(S): Compared with fibroblasts from normal peritoneum, fibroblasts from adhesions had significant complementary decreases in apoptosis and an increase in proliferation. In response to hypoxia, the Bcl-2-bax mRNA ratio was significantly higher in fibroblasts from adhesions as compared with fibroblasts from normal peritoneum. The degree of apoptosis induced by hypoxia correlated with the increase in the bcl-2-bax ratio in fibroblasts derived from both normal peritoneum and adhesions. CONCLUSION(S): Hypoxia induces proliferation while inhibiting apoptosis in fibroblasts from adhesion, thereby creating a phenotype that would favor adhesion development. PMID- 12095504 TI - Hypoxia-induced irreversible up-regulation of type I collagen and transforming growth factor-beta1 in human peritoneal fibroblasts. AB - OBJECTIVE: To determine whether restoration of normoxia after a hypoxic insult returns the molecular expression of type I collagen and TGF-beta1 to baseline levels. DESIGN: Prospective experimental study. SETTING: University medical center. PATIENT(S): Primary cultures of fibroblasts established from peritoneal tissues of five patients. INTERVENTION(S): Hypoxia treatment of the primary cultured fibroblasts. MAIN OUTCOME MEASURE(S): Cultured human peritoneal fibroblasts (HPF) were maintained under hypoxic conditions (2% oxygen) for 24 hours and then transferred into normal culture conditions (normoxia) for another 24 hours. Total cellular RNA from cells was collected and subjected to multiplex reverse transcription polymerase chain reaction to quantitate type I collagen and transforming growth factor (TGF)-beta1 mRNA levels in response to these treatments. RESULT(S): Hypoxia treatment resulted in 30% and 50% increases in type I collagen and TGF-beta1 expression, respectively. Restoration of normoxia after hypoxia treatment failed to restore type I collagen and TGF-beta1 expression to their baseline levels. CONCLUSION(S): These data support the hypothesis that hypoxia induces irreversible molecular changes in peritoneal fibroblasts that produce a phenotype that increases extracellular matrix expression and thereby would promote adhesion development. Thus once a phenotype consistent with increased adhesion development is manifested, restoration of oxygen supply does not reverse the stimulation of HPF type I collagen and TGF beta1 expression. This observation may in part explain the clinical observation that adhesion reformation is more difficult to prevent than de novo adhesion formation. PMID- 12095506 TI - Transforming growth factor-beta1 modulates expression of adhesion and cytoskeletal proteins in human peritoneal fibroblasts. AB - OBJECTIVE: To determine the effects of TGF-beta1 on the expression of the alpha1, alpha2, alpha5, alpha(v), and alpha6 integrin subunits and on vinculin, and F actin in human peritoneal fibroblasts. DESIGN: Descriptive study using cell culture, reverse transcriptase-polymerase chain reaction (RT-PCR), and immunofluorescent and confocal microscopy. SETTING: Academic medical center. PATIENT(S): Gynecological surgery patients. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Effects of TGF-beta1 on the steady state levels of alpha5, alpha(v), and alpha6 integrin transcripts were examined in the normal peritoneal fibroblasts using RT-PCR. Expression levels of the alpha1, alpha2, alpha5, alpha(v), and alpha6 integrin subunits and of F-actin were measured by immunofluorescent microscopy. The distribution pattern of the integrin subunits, vinculin, and F-actin were examined using confocal microscopy. RESULT(S): TGF beta1 significantly up-regulated the expression levels of the alpha5, alpha(v), and alpha6 integrin subunits and modulated their expression pattern, resulting in relatively higher levels of these subunits in the focal contacts of peritoneal fibroblasts. It allocated vinculin expression primarily to the focal contacts of cells and caused distortion of F-actin structure. The transcript levels of the alpha5, alpha(v), and alpha6 integrin subunits were not altered by TGF-beta1 treatment. CONCLUSION(S): TGF-beta1 may promote postoperative adhesion formation by inducing the migration of peritoneal fibroblasts by altering the expression levels and patterns of specific integrin subunits, vinculin, and F-actin. PMID- 12095505 TI - Expression of vascular endothelial growth factor and thrombospondin-1 mRNA in patients with endometriosis. AB - OBJECTIVE: To investigate the expression of vascular endothelial growth factor (VEGF) mRNA and thrombospondin-1 (TSP-1) mRNA in endometriosis. DESIGN: Molecular studies in human tissue. SETTING: Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, P. R. China. PATIENT(S): Patients undergoing laparoscopy for infertility or other benign gynecologic conditions. INTERVENTION(S): Biopsies were taken from endometriotic lesions (red peritoneal lesion, ovarian endometrioma, and unterosacral ligament nudule) and eutopic endometrium during laparoscopy. MAIN OUTCOME MEASURE(S): mRNA expression from endometriotic lesion and eutopic endometrium was analyzed by reverse transcriptase polymerase chain reaction (PCR) and Northern blotting. RESULT(S): Among the endometriotic lesions, red peritoneal lesions expressed higher levels of VEGF mRNA and lower levels of TSP-1 mRNA, whereas ovarian endometrioma expressed lower levels of VEGF mRNA and higher levels of TSP-1 mRNA. Eutopic endometrium of women with endometriosis had higher expression levels of VEGF mRNA and lower expression levels of TSP-1 mRNA than that of women without endometriosis. CONCLUSION(S): The expression of VEGF and TSP-1 in endometriotic lesions appears to be associated with the extent of their neovascularization. The imbalance in expression of VEGF and TSP-1 in the endometrium may play a role in the development of endometriosis. PMID- 12095507 TI - Regulation of insulin-like growth factor binding protein secretion by human granulosa luteal cells in a polycystic ovary-like environment. AB - OBJECTIVE: To determine the effect of androstenedione (A), insulin, and LH on secretion of insulin-like growth factor binding proteins (IGFBPs) from human granulosa luteal cells. DESIGN: Human granulosa cells were cultured for a total of 4 days in serum-free medium containing A (10(-6) mol/L), with or without insulin (100 microU/mL-800 microU/mL), LH (1 microU/mL-10 microg/L), and A (10( 5) mol/L). SETTING: Granulosa cells were obtained from IVF procedures. PATIENT(S): Women undergoing IVF for tubal disease. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Immunoassay and autoradiographs of Western ligand blotting detected IGFBP accumulations in the medium. RESULT(S): Cultured granulosa cells secreted IGFBP-1 through IGFBP-4. Insulin (100 microU/mL-800 microU/mL), LH (1 microg/L-10 microg/L), and A (10(-5) mol/L) caused a significant decrease in IGFBP-1 accumulation in the medium both alone and when added in combination. The release of IGFBP-2 and IGFBP-4 was significantly stimulated by insulin, whereas LH had no effect. Elevated levels of androgen (10(-5) mol/L) significantly stimulated the secretion of IGFBP-2, whereas the release of IGFBP-4 was reduced. CONCLUSION(S): These results demonstrate that androgen and insulin are important regulators of IGFBP release and that elevated levels of the two hormones may contribute to the altered IGFBP profile found in PCOS follicles, compared with the case of estrogen-dominant follicles. PMID- 12095508 TI - Effectiveness of photodynamic ablation for destruction of endometrial explants in a rat endometriosis model. AB - OBJECTIVE: To evaluate the potential of photodynamic therapy with aminolevulinic acid (ALA-PDT) for ablation of endometrial explants in a rat endometriosis model and to compare the effect of ALA-PDT, electrosurgery, and surgical resection on normal peritoneum. DESIGN: Prospective controlled experimental trial. SETTING: University medical center. ANIMAL(S): Mature Sprague-Dawley female rats. INTERVENTION(S): Induction of endometriosis and subsequent treatment with ALA PDT; electrosurgery, and simple resection, and ALA-PDT of normal peritoneum. MAIN OUTCOME MEASURE(S): Histopathological assessment. RESULT(S): Systemic ALA followed by exposure to photoactivating light for 10 or 15 minutes resulted in ablation of all explants harvested 3-4 days after treatment. Permanent destruction was confirmed by absence of regrowth by week 3. Exposure of normal peritoneum to ALA-PDT resulted in initial necrosis, with complete recovery by day 16. Adhesions were present on day 16 in 50% of cases after electrosurgery and in 100% of cases after resection. No adhesions were present in ALA-PDT-treated animals. CONCLUSION(S): Systemic ALA followed by exposure to photoactivating light at relatively low power densities for periods as brief as 10 minutes resulted in ablation of endometriotic explants. Exposure of normal peritoneum to ALA-PDT resulted in complete resurfacing. Both electrosurgery and surgical resection resulted in a greater incidence of surface adhesions. PMID- 12095509 TI - Differential diagnosis of septate and bicornuate uterus by sonohysterography eliminates the need for laparoscopy. AB - OBJECTIVE: To determine whether sonohysterography can differentiate septate from bicornuate uterus. DESIGN: Prospective study. SETTING: Outpatient infertility clinic of a university hospital. PATIENT(S): Twenty patients with history of recurrent pregnancy loss and hysterosalpingographic diagnosis of septate or bicornuate uterus. INTERVENTION(S): Sonohysterography was done in all patients. Laparoscopy was performed to confirm diagnosis in 10 patients and was not done in 10 patients. All patients with septate uterus underwent hysteroscopic metroplasty; simultaneous laparoscopy was done in women who had diagnostic laparoscopy. Abdominal metroplasty was performed in patients with bicornuate uterus. MAIN OUTCOME MEASURE(S): Shape of the uterine cavity after surgery and pregnancy outcome. RESULT(S): The shape of the uterine cavity was normal in seven cases in each group and almost normal in six other patients after surgery. There were two pregnancy losses after metroplasty, one in a woman who had laparoscopy and one in a woman who did not have laparoscopy. One case of bicornuate uterus occurred in each group. CONCLUSION(S): Sonohysterography can differentiate septate and bicornuate uterus and may eliminate the need for laparoscopy to diagnose these uterine anomalies. PMID- 12095510 TI - Laser-assisted hatching of embryos is better than the chemical method for enhancing the pregnancy rate in women with advanced age. AB - OBJECTIVE: Assisted hatching may enhance embryo implantation. This study was conducted to examine the efficacy of the laser- and chemical-assisted hatching for promotion of implantation (IR), pregnancy (PR), and delivery rate (DR) in older women undergoing IVF cycles. DESIGN: Prospective study. SETTING: An IVF unit of a medical center. PATIENT(S): A total of 601 embryos from 141 women aged > or =38 years underwent controlled ovarian hyperstimulation (COH) and assisted hatching. INTERVENTION(S): The study population was divided into two groups: group 1 had laser-assisted hatching (n = 85) and group 2 had chemical-assisted hatching (n = 56). Before the transfer, the day 3 embryos were hatched by using a 1.48-microm noncontact diode laser or acid Tyrode's solution. MAIN OUTCOME MEASUREMENT(S): The IR, PR, and DR between the groups were compared. RESULT(S): There were no statistical differences between groups in age, E2 concentrations during hCG administration, gonadotrophin dosage, embryo grade, the numbers of oocytes retrieved, oocytes fertilized, and embryos transferred. Higher IR, PR, and DR were noted in the laser-assisted hatching group. The IR, PR, and DR were: group 1, 8.2%/31.8%/24.7% and group 2, 3.8%/16.1%/10.7%, respectively. CONCLUSION(S): Laser-assisted hatching of embryos is more effective than the chemical method in enhancing the IR and PR of women with advanced age. The laser system allows an easier, faster, and safer micromanipulation of the zona pellucida, which provided a better method in zona drilling. PMID- 12095511 TI - Pulmonary endometriosis in a patient with unicornuate uterus and noncommunicating rudimentary horn. AB - OBJECTIVE: To report a rare case of a patient with catamenial hemoptysis, secondary infertility, and endometriosis associated with a unicornuate uterus and noncommunicating rudimentary horn. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 29-year-old woman who developed progressive catamenial hemoptysis and secondary infertility was evaluated at the University Hospital of Crete. INTERVENTION(S): The complete history, laboratory data, laparoscopic findings, and chest magnetic resonance image of this patient were analyzed. A GnRH agonist, leuprolide acetate, was successfully administered. MAIN OUTCOME MEASURE(S): Diagnosis and appropriate treatment of pulmonary endometriosis in a patient with rudimentary uterine horn. RESULT(S): Treatment with a GnRH agonist achieved suppression of both menstruation and hemoptysis. After 6 months of normal menstrual activity, the symptoms reappeared. The patient was again treated with leuprolide acetate (3.75 mg/mo IM) for 6 months and remained asymptomatic. In fact, the patient became pregnant after cessation of therapy. Finally, the patient was treated successfully with removal of the rudimentary uterine horn during cesarean section. Three-year follow-up showed disappearance of the chest symptoms. CONCLUSION(S): Pulmonary endometriosis and unicornuate uteri are rare. To our knowledge, this is the first case of catamenial hemoptysis with a congenital mullerian anomaly. We describe successful management using a combination of GnRH agonist and surgical resection of the rudimentary uterine horn. PMID- 12095512 TI - A comparative genomic hybridization study in a 46,XX male. AB - OBJECTIVE: To identify Y chromosome material in an azoospermic male with an XX karyotype. DESIGN: Case report. SETTING: Faculty of medicine and Centro de Patologia Celular (CPC) medical center. PATIENT(S): A 33-year-old man with infertility. INTERVENTION(S): G-banding, fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR), and comparative genomic hybridization (CGH). MAIN OUTCOME MEASURE(S): FISH for X and Y chromosomes, PCR for the SRYgene and amelogenin gene in the Xp (AMGX) and (AMGY), and losses or gains with CGH. RESULT(S): FISH analysis using X and Y chromosome-specific probes showed an X chromosome containing Y chromosome sequences on the top of the short arm; this Y chromosome region was not visible by conventional cytogenetic analysis. PCR amplification of DNA showed the presence of the sex-determining region of the Y chromosome (SRY) and the amelogenin gene in the pseudoautosomal boundary of the X chromosome (AMGX). CGH confirmed the presence of the chromosome region Yp11.2 pter and detected the presence of the two otherwise normal X chromosomes. CONCLUSION(S): The two Xpter (XPAR1) pseudoautosomal regions present in this XX male suggest the need to reevaluate XX males using CGH and PCR to characterize the clinical variability in XX males due to genes other than those located on the Y chromosome. PMID- 12095513 TI - Transabdominal-transperitoneal ultrasound-guided oocyte retrieval in a patient with mullerian agenesis. AB - OBJECTIVE: To describe the approach of transabdominal-transperitoneal ultrasound guided oocyte retrieval undertaken in a patient with mullerian agenesis and ovarian malposition who underwent a successful gestational carrier treatment cycle. DESIGN: Case report. SETTING: A tertiary-referral reproductive medicine unit. PATIENT(S): A 26-year-old patient with mullerian agenesis. Her 44-year-old mother served as the gestational carrier. INTERVENTION(S): Controlled ovarian hyperstimulation, transabdominal-transperitoneal ultrasound-guided oocyte retrieval, embryo transfer. MAIN OUTCOME MEASURE(S): Oocyte recovery rate, fertilization rate, pregnancy test. RESULT(S): Six oocytes were retrieved using a percutaneous transperitoneal needle puncture under ultrasound guidance. Two cleavage-stage embryos were transferred to the gestational carrier, resulting in a twin pregnancy. CONCLUSION(S): For various reasons, patients with mullerian agenesis may not be candidates for standard transvaginal ultrasound-guided oocyte retrieval. Although laparoscopic oocyte retrieval has been frequently used in this setting, the approach of transabdominal-transperitoneal ultrasound-guided oocyte retrieval may offer further advantages in select cases. PMID- 12095514 TI - Nonrobertsonian translocation t(6;11) is associated with infertility in an oligoazoospermic male. AB - OBJECTIVE: To analyze an unusual nonrobertsonian translocation t(6;11) found in an infertile, oligoazoospermic, but otherwise apparently healthy man. DESIGN: Case report with review of the scientific literature. SETTING: Academic research environment. PATIENT(S): Infertile, oligoazoospermic, but otherwise apparently healthy man. INTERVENTION(S): Peripheral blood lymphocytes were obtained for karyotyping, and metaphases were studied by standard GBG, RBG, and CBG banding procedures. MAIN OUTCOME MEASURE(S): Sperm count, morphology, and GBG, RBG, and CBG banding. RESULT(S): A karyogram revealed a nonrobertsonian balanced translocation t(6;11) with breakpoints at 6q15 and 11p15, as shown by R-, G-, and C-banding. CONCLUSION(S): Physical contact of unpaired autosomal material with sex chromosomes, which leads to spermatogenic arrest, may be one of the factors that accounts for infertility in men with autosomal aberrations. PMID- 12095515 TI - Cyclical expression of the DNA mismatch repair enzyme hMLH1 in normal endometrium. PMID- 12095516 TI - Ovarian reserve after uterine artery embolization for leiomyomata. PMID- 12095517 TI - Use of infrared functional imaging to detect impaired thermoregulatory control in men with asymptomatic varicocele. PMID- 12095518 TI - Impact of cryopreservation and subsequent embryo transfer on the outcome of in vitro fertilization in patients at high risk for ovarian hyperstimulation syndrome. PMID- 12095519 TI - The importance of acknowledged error. PMID- 12095521 TI - Appropriate credit. PMID- 12095523 TI - The purity of granulosa cell preparation? PMID- 12095525 TI - Surgical management of endometriomas--compromised ovarian function? PMID- 12095531 TI - Aging and infertility in women: a committee opinion. PMID- 12095532 TI - Inhibitory effects of 1,4-DHP antagonists on synaptic GABA release modulated by BAY-K 8644 in mechanically dissociated rat substantia innominata. AB - The effects of dihydropyridine (1,4-DHP) agonist and antagonists on miniature inhibitory postsynaptic currents (mIPSCs) were investigated in mechanically dissociated rat substantia innominata neurons attached to native GABAergic presynaptic nerve terminals, namely 'synaptic bouton preparation', using nystatin perforated patch recording mode under voltage-clamp conditions. BAY-K 8644 (BAY K), an L-type Ca(2+) channel agonist, reversibly and concentration dependently facilitated the GABAergic mIPSC frequency without altering the distribution of current amplitudes. Removal of extracellular Ca(2+) completely suppressed the facilitatory effect of BAY-K on mIPSC frequency. The facilitatory effect of BAY-K on mIPSC frequency was maintained even in the presence of selective N-, P- and Q type Ca(2+) channel antagonists, such as 3 x 10(-6) M omega-conotoxin-GVIA (omega CgTX-GVIA), 3 x 10(-8) M omega-agatoxin-IVA (omega-AgTX-IVA) and 3 x 10(-6)M omega-conotoxin-MVIIC (omega-CmTX-MVIIC). However, nicardipine (3 x 10(-6) M) and nimodipine (3 x 10(-6) M), 1,4-DHP antagonists, significantly inhibited the mIPSC frequency enhanced by BAY-K by 37 +/- 5 and 42 +/- 6%, respectively. These results suggest the possible existence of L-type Ca(2+) channels in GABAergic presynaptic nerve terminals. PMID- 12095533 TI - Nipradilol induces vasodilation of canine isolated posterior ciliary artery via stimulation of the guanylyl cyclase-cGMP pathway. AB - We examined the effect of nipradilol on contraction of the posterior ciliary artery induced by high potassium or norepinephrine and on cyclic GMP (cGMP) levels in the posterior ciliary artery of dogs. Nipradilol caused dose-dependent relaxation of KCl-and norepinephrine-induced contractions of posterior ciliary artery. The relaxant effect of nipradilol on norepinephrine-contracted ciliary artery was significantly greater than that on KCl-contracted ciliary artery. In KCl-contracted ciliary artery, N(G)-nitro-L-arginine methyl ester hydrochloride (L-NAME, 10(-4) M) did not alter the relaxant effect of nipradilol, whereas 1H 1,2,4-oxadiazolo-4,3-a-quinoxalin-1-one (ODQ, 10(-6) M) significantly inhibited this effect. Ethacrynic acid at 10(-5) M, sulfasalazine at 10(-4) M and S decylglutathione at 10(-4) M (glutathione S-transferase inhibitors) did not inhibit the relaxant effect of nipradilol. In addition, nipradilol produced dose dependent increases in cGMP levels in the canine posterior ciliary artery. These findings indicate that nipradilol-induced vasorelaxation in the canine posterior ciliary artery occurs via stimulation of the guanylyl cyclase-cGMP pathway. PMID- 12095534 TI - Effects of maternal deprivation on adrenal and behavioural responses in rats with anterodorsal thalami nuclei lesions. AB - There is evidence that repeated maternal isolation of neonatal rats may influence both emotional behavior and Hypothalamic-Pituitary Adrenal (HPA) activity. On the other hand the Anterodorsal Thalami Nuclei (ADTN) exerts an inhibitory influence on the hypophyso-adrenal system under basal and stressful conditions. In the present work we investigated whether neonatal maternal deprivation produces long term effects on the ADTN regulation of behavioral patterns (open field test) and on HPA axis activity. Specifically, we sought to determine whether adult female rats with ADTN lesions, previously isolated for 4.5 hours daily during the first 3 weeks of life, react in endocrinologically and behaviourally distinct manner as compared to controls. The examined groups were: non maternally deprived (NMD)/sham lesioned, NMD/lesioned, maternally deprived (MD)/sham lesioned, MD/lesioned with and without the open field test. At 3 months MD/sham lesioned animals showed a marked decrease in ambulation (P < 0.01), and with ADTN lesion, the rearing values were lower (P < 0.01) and grooming higher (P < 0.05) than NMD. This last data would indicate a high emotional index. Regarding the activity of the HPA axis, maternal deprivation induced a significant decrease in plasma ACTH concentration both in sham and lesioned animals (P < 0.001), and plasma Corticosterone (C) increased in sham animals (P < 0.001). This data would indicate a higher sensitivity of the adrenal glands. After the open field test ACTH and C were different between deprived and non-deprived animals depending on the ADTN lesion. Taking into consideration the increase of ACTH levels in sham lesioned MD animals exposed to the test, we could conclude that this new situation was a stressful situation. Finally in the present work, it was very difficult to relate the behavioral parameters with the endocrine data. It is known that depending on the context, corticosteroids may produce opposite effects on emotional behavior via different receptors in the brain.In summary, neonatal maternal deprivation induced alterations of behavioral patterns and affected the ADTN inhibitory influence on ACTH and C secretion. PMID- 12095535 TI - Role of D and E domains in the migration of vascular smooth muscle cells into fibrin gels. AB - The structure of fibrin plays an important role in the organization of thrombi, the development of atherosclerosis, and restenosis after PTCA. In this study, we examined the mechanisms of the migration of vascular smooth muscle cells (SMCs) into fibrin gels, using an in vitro assay system. Cultured SMCs from bovine fetal aortic media migrated into fibrin gels prepared with thrombin, which cleaves both fibrinopeptides A and B from fibrinogen, without other chemotactic stimuli. Both desA fibrin gels prepared with batroxobin, which cleaves only fibrinopeptide A, and desB fibrin gels prepared with Agkistrodon contortrix thrombin-like enzyme (ACTE), which cleaves only fibrinopeptide B, similarly induced the migration of SMCs compared to fibrin gels prepared with thrombin. These results suggest that the cleavage of fibrinopeptides is not necessary, but rather that the three dimensional structure of the gel may be important for the migration of SMCs. Furthermore, gels prepared with protamine sulfate, which forms fibrin-like gels non-enzymatically, similarly induced the migration of SMCs compared to the gels prepared with thrombin. Both anti-fibrin(ogen) fragment D and anti-fibrin(ogen) E antibodies inhibited the migration of SMCs into fibrin gels, suggesting that both the D and E domains of fibrin(ogen) are involved in the migration of SMCs into fibrin gels. The addition of GRGDS, a synthetic RGD-containing peptide, but not that of GRGES, a control peptide, partially inhibited the migration of SMCs into fibrin gels, suggesting that the migration of SMCs into fibrin gels is at least in part dependent on the RGD-containing region of the alpha chain. The migration of SMCs into fibrin gels was also inhibited by a monoclonal antibody for integrin alpha v beta 3 and alpha 5 beta 1, indicating that migration is dependent on these integrins. Furthermore, both fibrin(ogen) fragments D and E inhibited the migration of SMCs into fibrin gels, suggesting that these fragments, generated during fibrino(geno)lysis, may be relevant in the regulation of SMC migration into fibrin gels. PMID- 12095536 TI - The effect of grapefruit juice and seville orange juice on the pharmacokinetics of dextromethorphan: the role of gut CYP3A and P-glycoprotein. AB - The objective of this study was to determine the effects of grapefruit juice and seville orange juice on dextromethorphan (DM) pharmacokinetics. Eleven healthy volunteers were studied over a 3-week period consisting of 5 study days each separated by a three-day washout. All subjects refrained from drinking caffeine containing beverages (coffee, soda, etc.) 8 h before orally taking DM (30 mg) with 200 ml water, 200 ml grapefruit juice, 200 ml water, 200 ml seville orange juice, and 200 ml water on Study Days 1 to 5. Aliquots of urine samples were assayed and analysed for DM, and the DM metabolites dextrorphan, 3 methoxymorphinan and 3-hydroxymorphinan using a validated HPLC method employing a phenyl column and a fluorescence detection. Results suggests that DM could provide some useful information on P-glycoprotein or related membrane efflux protein activity in the human gastro-intestinal tract. Bioavailability (F) of DM increased significantly with grapefruit and seville orange juice, but only returned to half the baseline value after three days of washout. This confirms that grapefruit and seville orange juice are long-lasting and perhaps irreversible inhibitors of gut CYP3A/P-glycoprotein. Grapefruit and seville orange juice appeared to have the same overall effect on DM pharmacokinetics. In addition, this paper presents a novel method of phenotyping for CYP2D6, CYP3A and P-glycoprotein using DM as a probe. PMID- 12095537 TI - Growth, morphological and biochemical changes in oxa-spermine derivative-treated MCF-7 human breast cancer cells. AB - The growth inhibitory properties of two oxa-spermine derivatives named compound 1 and compound 2, representatives of a novel type of polyamine derivatives, were studied. Dose-response growth inhibitory curves obtained after 48h drug exposure demonstrated the much higher cytotoxic activity of compound 1 towards MCF-7 human breast cancer cells. Further experiments with compound 1 showed that this oxa spermine derivative exhibited considerable cytotoxicity with IC(50) values of 3.74 microM and 2.93 microM after 24h and 48h drug exposure respectively. In MCF 7 cells, after 8h drug (10 microM) exposure it caused shrinkage, chromatin condensation and nuclear fragmentation. However, no clear DNA laddering was detected in treated cells. Drug treatment provoked an increase in polyamine oxidase (PAO) activity. This enzyme is able to produce cytotoxic H(2)O(2) and 3 acetamidopropanal, catalyzing the oxidative deamination of N(1)-acetylated derivatives of spermine and spermidine to spermidine and putrescine respectively. Taken together these data demonstrate that the novel oxa-polyamine derivative compound 1 has considerable cytotoxic activity towards MCF-7 cells and indicate that an induction of PAO may be involved in its cytotoxic and apoptotic effects. PMID- 12095538 TI - Upregulation of small GTPase RhoA in the basilar artery from diabetic (mellitus) rats. AB - The goal of this study was to determine whether RhoA, a small GTPase, might be involved in the development of cerebral pathogenesis in diabetes. Male SD rats (n = 120) were divided into six groups: diabetic for 2, 4, 8 weeks, and an age matched control group. Diabetes was induced by intravenous injection of streptozotocin (50 mg/kg). RhoA mRNA expression in basilar artery was measured by competitive RT-PCR. RhoA mRNA level was significantly increased in 4 weeks (184.1 +/- 28.5%, n = 7) and 8 weeks (218.7 +/- 24.5%, n = 7) after STZ injection compared to the age matched control basilar arteries (P < 0.05). Western blot was used to measure the membrane binding RhoA level to represent the activity of RhoA. We found that RhoA activity was strikingly increased in the diabetic basilar artery (n = 10 in each groups) compared to control basilar artery after STZ injection. Our data demonstrated that there was an upregulation of RhoA in the basilar artery of STZ induced diabetic rats, suggesting that RhoA might be involved in the cerebral vascular pathogenesis during diabetes mellitus. PMID- 12095539 TI - Ovarian follicular differentiation with prepubertal gonadotropin surges and gonadotropin priming in mice. AB - Preantral follicles were mechanically isolated from the ovaries of 1.5 to 8 week old mice and cultured in vitro for 4 days in the presence or absence of either activin A or FSH. Plasma gonadotropin, estradiol and immunoreactive (IR) inhibin levels were measured. Cultured follicles showed stepwise changes in response to recombinant human (rh) FSH, with no response until 11 days, a gradual increase from 2 weeks, culminating in a strong response to rhFSH at 8 weeks. The response to activin A was vice versa. It enhanced the effect of rhFSH on preantral follicular growth of up to 4-week-old mice, but inhibited the effect of rhFSH in 8-week-old mice. The peak of the prepubertal gonadotropin surge was observed on day 11. Seven-day-old mice were treated with either luteinizing hormone releasing hormone (LHRH) or rhFSH or human chorionic gonadotropin (hCG) for 3 consecutive days from day 7, and follicles were collected on day 11. Those follicles showed enhanced response to rhFSH, no response to activin A, and an enhanced response to the combination of rhFSH and activin A, suggesting that the chronological changes in follicular response are a result of the prepubertal gonadotropin surge. PMID- 12095540 TI - Activation of CFTR Cl(-) channel by tyrphostins via a protein tyrosine kinase independent pathway in forskolin-stimulated renal epithelial A6 cells. AB - We studied effects of tyrphostin A23 (an inhibitor of protein tyrosine kinase; PTK) and tyrphostin A63 (an inactive analog of tyrphostin A23) on forskolin activated cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channels and Cl(-) secretion in renal epithelial A6 cells. Tyrphostin A23 and A63 had no effects on the basal CFTR Cl(-) channel and Cl(-) secretion. However, under the forskolin-stimulated condition, tyrphostin A23 and A63 stimulated Cl(-) secretion by activating CFTR Cl(-) channels. These observations suggest that: 1) tyrphostin A23 and A63 stimulate the cAMP-activated CFTR Cl(-) channel via a PTK independent, structure-dependent mechanism, and 2) tyrphostin A23 and A63 do not stimulate the basal CFTR Cl(-) channel. These lead us to an idea that: 1) cAMP might cause a conformational change of CFTR Cl(-) channel which is accessible by tyrphostins, and 2) tyrphostins would stimulate translocation of the cAMP modified channel to the apical membrane by binding to the channel. PMID- 12095541 TI - The response of the lamb ductus arteriosus to endothelin: developmental changes and influence of light. AB - Endothelin-1 (ET-1) is a putative messenger of oxygen in the ductus arteriosus. Since the ability of the vessel to contract to oxygen increases with gestation, we wished to ascertain whether ET-1 action is also developmentally regulated. A corollary objective was to assess whether any gestational variation in the ET-1 contraction is due to a change in the ET(A)-mediated action or to a shift in the balance between opposing, contractile (ET(A) - mediated) and relaxant (ET(B) mediated), actions. Experiments were performed with isolated ductal strips from preterm (0.7 gestation) and near-term fetal lambs. ET-1 contracted the ductus dose-dependently (10(-10)-10(-7) M) at both ages; however, the peak contraction was about double in magnitude at term. Regardless of age, ET-1 contraction was greater with preparations kept in the dark compared to those exposed to light. This effect of light was not seen after removing the endothelium or when treating the intact tissue with the ET(B) antagonist BQ788 (1 microM). In the dark, however, BQ788 did not modify significantly the ET-1 response at either age. We conclude that ET-1 becomes a stronger ductus constrictor with fetal age, conceivably by acting on ET(A) receptors. Hence, the concept of ET-1 mediating the oxygen contraction is further validated. Peculiarly, the ET-1 contraction is curtailed by light through a hitherto undefined ET(B) receptor-linked process. PMID- 12095543 TI - Management of supraglottic carcinoma. PMID- 12095542 TI - Intra- and inter-individual relationships between central and peripheral serotonergic activity in humans: a serial cerebrospinal fluid sampling study. AB - Data are lacking concerning the longitudinal covariability and cross-sectional balance between central and peripheral 5-HIAA concentrations in humans and on the possible associations between tobacco smoking or post-traumatic stress disorder (PTSD) and CSF and plasma 5-HIAA concentrations. Using serial cerebrospinal fluid (CSF) and blood sampling, we determined the concentrations of 5-HIAA in CSF and plasma over 6 h, and examined their relationships in healthy volunteers and patients with PTSD-both smokers and nonsmokers. Patients with PTSD and healthy volunteers had very similar CSF 5-HIAA concentrations. Significant and positive correlations between CSF and plasma 5-HIAA levels were observed within individuals, but this CNS-peripheral 5-HIAA relationship was significantly reduced in smokers (nonsmokers: mean r = 0.559 +/- 0.072; smokers: mean r = 0.329 +/- 0.064 p < 0.038). No significant cross-sectional, interindividual correlation of mean CSF and mean plasma 5-HIAA was seen (r = 0.094). These data show that changes in CSF 5-HIAA levels within an individual over time are largely reflected in plasma 5-HIAA, albeit significantly less so in smokers. The present results therefore suggest that clinically, longitudinal determination of plasma 5-HIAA concentrations within an individual patient can be used to make inferences about relative changes in integrated CSF 5-HIAA concentrations. However, plasma 5-HIAA concentrations provide no significant information about absolute levels of the serotonin metabolite in the CSF. PMID- 12095544 TI - Contributions of L. H. Gray to radiation physics, biology, and oncology. PMID- 12095545 TI - The Gray Lecture 2001: coming technical advances in radiation oncology. AB - PURPOSE: To review the current limits on the efficacy of radiotherapy (RT) due to technical factors and to assess the potential for major improvements in technology. METHODS AND MATERIALS: The method of this review was to assess the efficacy of current RT in general terms; strategies for improving RT; historical record of technological advances; rationale for further reductions of treatment volume; and importance of defining and excluding nontarget tissues from the target volume. The basis for the interest in proton beam RT is developed, and the relative dose distributions of intensity-modulated radiotherapy (IMRT) and intensity-modulated proton RT (IMPT) are discussed. The discovery of the proton and the first proposal that protons be used in RT is described. This is followed by a brief mention of the clinical outcome studies of proton RT. Likely technical advances to be integrated into advanced proton RT are considered, specifically, four-dimensional treatment planning and delivery. Finally, the increment in cost of some of these developments is presented. RESULTS: For definitive RT, dose limits are set by the tolerance of normal tissues/structures adjacent or near to the target. Using imaging fusion of CT, MRI, positron emission tomography, magnetic resonance spectroscopic imaging, and other studies will result in improved definition of the target margins. Proton beams are likely to replace photon beams because of their physical characteristics. Namely, for each beam path, the dose deep to the target is zero, across the target it is uniform, and proximal to the target it is less. Proton therapy can use as many beams, beam angles, noncoplanar, and dynamic, as well as static, intensity modulation, as can photon plans. The ability for much greater accuracy in defining the target position in space and then maintaining the target in a constant position in the radiation beam despite target movement between and during dose fractions will be possible. The cost of proton RT will be modestly higher than comparable high technology photon therapy. CONCLUSION: The technology of RT is clearly experiencing intense and rapid technical developments as pertains to treatment planning and dose delivery. It is predicted that radical dose RT will move to proton beam technology and that the treatment will be four dimensional (the fourth dimension is time). The impact will be higher tumor control probability and reduced frequency and severity of treatment-related morbidity. PMID- 12095546 TI - Analysis of radiation-induced liver disease using the Lyman NTCP model. AB - PURPOSE: To describe the dose-volume tolerance for radiation-induced liver disease (RILD) using the Lyman-Kutcher-Burman (LKB) normal tissue complication probability (NTCP) model. METHODS AND MATERIALS: A total of 203 patients treated with conformal liver radiotherapy and concurrent hepatic arterial chemotherapy were prospectively followed for RILD. Normal liver dose-volume histograms and RILD status for these patients were used as input data for determination of LKB model parameters. A complication was defined as Radiation Therapy Oncology Group Grade 3 or higher RILD < o r =4 months after completion of radiotherapy. A maximal likelihood analysis yielded best estimates for the LKB NTCP model parameters for the liver for the entire patient population. A multivariate analysis of the potential factors associated with RILD was also completed, and refined LKB model parameters were obtained for patient subgroups with different risks of RILD. RESULTS: Of 203 patients treated with focal liver irradiation, 19 developed RILD. The LKB NTCP model fit the complication data for the entire group. The "n" parameter was larger than previously described, suggesting a strong volume effect for RILD and a correlation of NTCP with the mean liver dose. No cases of RILD were observed when the mean liver dose was <31 Gy. Multivariate analysis demonstrated that in addition to NTCP and the mean liver dose, a primary hepatobiliary cancer diagnosis (vs. liver metastases), bromodeoxyuridine hepatic artery chemotherapy (vs. fluorodeoxyuridine chemotherapy), and male gender were associated with RILD. For 169 patients treated with fluorodeoxyuridine, the refined LKB model parameters were n = 0.97, m = 0.12, tolerance dose for 50% complication risk for whole organ irradiated uniformly [TD50(1)] = 45.8 Gy for patients with liver metastases, and TD50(1) = 39.8 Gy for patients with primary hepatobiliary cancer. CONCLUSION: These data demonstrate that the liver exhibits a large volume effect for RILD, suggesting that the mean liver dose may be useful in ranking radiation plans. The inclusion of clinical factors, especially the diagnosis of primary hepatobiliary cancer vs. liver metastases, improves the estimation of NTCP over that obtained solely by the use of dose-volume data. These findings should facilitate the application of focal liver irradiation in future clinical trials. PMID- 12095547 TI - Precise and real-time measurement of 3D tumor motion in lung due to breathing and heartbeat, measured during radiotherapy. AB - PURPOSE: In this work, three-dimensional (3D) motion of lung tumors during radiotherapy in real time was investigated. Understanding the behavior of tumor motion in lung tissue to model tumor movement is necessary for accurate (gated or breath-hold) radiotherapy or CT scanning. METHODS: Twenty patients were included in this study. Before treatment, a 2-mm gold marker was implanted in or near the tumor. A real-time tumor tracking system using two fluoroscopy image processor units was installed in the treatment room. The 3D position of the implanted gold marker was determined by using real-time pattern recognition and a calibrated projection geometry. The linear accelerator was triggered to irradiate the tumor only when the gold marker was located within a certain volume. The system provided the coordinates of the gold marker during beam-on and beam-off time in all directions simultaneously, at a sample rate of 30 images per second. The recorded tumor motion was analyzed in terms of the amplitude and curvature of the tumor motion in three directions, the differences in breathing level during treatment, hysteresis (the difference between the inhalation and exhalation trajectory of the tumor), and the amplitude of tumor motion induced by cardiac motion. RESULTS: The average amplitude of the tumor motion was greatest (12 +/- 2 mm [SD]) in the cranial-caudal direction for tumors situated in the lower lobes and not attached to rigid structures such as the chest wall or vertebrae. For the lateral and anterior-posterior directions, tumor motion was small both for upper- and lower-lobe tumors (2 +/- 1 mm). The time-averaged tumor position was closer to the exhale position, because the tumor spent more time in the exhalation than in the inhalation phase. The tumor motion was modeled as a sinusoidal movement with varying asymmetry. The tumor position in the exhale phase was more stable than the tumor position in the inhale phase during individual treatment fields. However, in many patients, shifts in the exhale tumor position were observed intra- and interfractionally. These shifts are the result of patient relaxation, gravity (posterior direction), setup errors, and/or patient movement.The 3D trajectory of the tumor showed hysteresis for 10 of the 21 tumors, which ranged from 1 to 5 mm. The extent of hysteresis and the amplitude of the tumor motion remained fairly constant during the entire treatment. Changes in shape of the trajectory of the tumor were observed between subsequent treatment days for only one patient. Fourier analysis revealed that for 7 of the 21 tumors, a measurable motion in the range 1-4 mm was caused by the cardiac beat. These tumors were located near the heart or attached to the aortic arch. The motion due to the heartbeat was greatest in the lateral direction. Tumor motion due to hysteresis and heartbeat can lower treatment efficiency in real-time tumor tracking-gated treatments or lead to a geographic miss in conventional or active breathing controlled treatments. CONCLUSION: The real-time tumor tracking system measured the tumor position in all three directions simultaneously, at a sampling rate that enabled detection of tumor motion due to heartbeat as well as hysteresis. Tumor motion and hysteresis could be modeled with an asymmetric function with varying asymmetry. Tumor motion due to breathing was greatest in the cranial caudal direction for lower-lobe unfixed tumors. PMID- 12095548 TI - Influence of tumor volume on survival in patients irradiated for non-small-cell lung cancer. AB - PURPOSE: To investigate the importance of CT-defined total tumor volume (TTV) on overall survival (OS) in patients with unresectable or medically inoperable non small-cell lung carcinoma (NSCLC). METHODS AND MATERIALS: Between 1991 and 1998, 150 evaluable patients with Stage I-IIIB NSCLC were treated with three dimensionally planned conformal radiotherapy and curative intent at Duke University Medical Center. On the treatment-planning CT, the primary tumor and nodal volumes were identified and subsequently combined to form the TTV. The TTV was compared with the stage and outcome with respect to OS, local progression free survival, and distant failure-free survival using the Kruskall-Wallis analysis of variance and Kaplan-Meier actuarial method. To account for the potentially confounding effects of therapeutic and patient-specific covariates on survival, the Cox proportional hazard regression model was used. RESULTS: The TTVs in patients with Stage I disease (median 19 cm3) were smaller than in patients with Stage II (median 80 cm3) or Stage III (median 97 cm3; p <0.001) disease. The Stage II TTVs were not significantly different from those of Stage III (post-hoc test according to Bonferroni). Prolonged OS was independently associated with a small TTV (<80 vs. >80 cm3 [median]; p = 0.01), young age (<60 vs. > or =60 years; p = 0.03), high Karnofsky performance status (< o r =70 vs. >70; p = 0.04), and female gender (p = 0.04). Both stage (p = 0.7) and T stage (p = 0.06) were of less importance for OS than was the TTV, according to multivariate modeling. Increased local progression-free survival (p = 0.001) and distant failure-free survival (p = 0.03) were independently associated with a small TTV (i.e., <80 cm3). The results were unchanged if the TTV was analyzed as a continuous variable. CONCLUSION: A strong independent association between a small CT-defined TTV and prolonged survival in patients with NSCLC selected for curative/definitive RT was found. Future therapeutic studies in NSCLC should consider stratifying/adjusting for differences in TTV to avoid confounding effects on survival from variations in the TTV at baseline. PMID- 12095549 TI - Quality of life after curative radiotherapy in Stage I non-small-cell lung cancer. AB - PURPOSE: The aim of this study was to investigate changes in quality of life (QOL) among medically inoperable Stage I non-small-cell lung cancer (NSCLC) patients treated with curative radiotherapy. PATIENTS AND METHODS: The study sample was composed of 46 patients irradiated for Stage I NSCLC. Quality of life was assessed before, during, and after radiotherapy using the European Organization for the Research and Treatment of Cancer QLQ-C30 and QLQ-LC13. Changes in symptom and QOL scores over time were evaluated with a repeated measurement analysis of variance using the mixed effect modeling procedure, SAS Proc Mixed. Twenty-seven patients were treated only at the primary site, whereas for 19 patients, the regional lymph nodes were included in the target volume as well. RESULTS: The median follow-up time of patients alive was 34 months. The median survival was 19.0 months. None of the locally treated patients developed regional recurrence. A significant, gradual increase over time was observed for dyspnea, fatigue, and appetite loss. A significant, gradual deterioration was observed also for role functioning. No significant changes were noted for the other symptoms or the functioning scales. Significantly higher levels of dysphagia, which persisted up to 12 months, were observed in those in which the regional lymph nodes were treated, as compared to the locally treated patients. Radiation-induced pulmonary changes assessed with chest radiograph were more pronounced in the group treated with locoregional radiotherapy. CONCLUSIONS: After curative radiotherapy for Stage I medically inoperable NSCLC, a gradual increase in dyspnea, fatigue, and appetite loss, together with a significant deterioration of role functioning, was observed, possibly because of pre existing, slowly progressive chronic obstructive pulmonary disease and radiation induced pulmonary changes. Taking into account the low incidence of regional recurrences after local irradiation, the higher incidence and severity of radiation-induced changes, and the higher levels of dysphagia persisting up to 12 months, local irradiation of the primary tumor without elective irradiation of the regional lymph nodes may be the most appropriate treatment for patients with small, peripherally located tumors. PMID- 12095550 TI - Expression of hypoxia-inducible factor-1alpha in cervical carcinomas: correlation with tumor oxygenation. AB - PURPOSE: To investigate the relations between hypoxia-inducible factor-1 (HIF-1), tumor oxygenation, and clinical correlates in patients with locally advanced carcinoma of the uterine cervix. METHODS AND MATERIALS: Biopsies from 42 patients with invasive cervical carcinoma and previous polarographic O2 measurements were assessed for the expression of HIF-1alpha using digitized microscopic imaging and analysis. RESULTS: The HIF-1alpha expression levels ranged from <0.1% to 10.7% of the total tumor area; the positive staining was localized exclusively to the nuclei. Three distinct arrangement patterns of HIF-1alpha-positive cells in relation to blood vessels were identified using spatial image mapping: (1) most HIF-1alpha-positive cells were located within the typical oxygen diffusion distance in tissue (< or =150 microm to the nearest blood vessel); (2) most HIF 1alpha-positive cells were located in the vicinity (< or =60 microm) of the blood vessels; and (3) no apparent spatial relationship was found between HIF-1alpha positive cells and blood vessels. A statistically significant association was found between HIF-1alpha expression and tumor oxygenation (Spearman correlation coefficient = 0.4, p <0.01), as determined with the Eppendorf pO2 histograph. No correlation was found between the level of HIF-1alpha expression and patient outcome, using disease-free survival as the end point. CONCLUSION: Our results suggest that HIF-1alpha expression may represent a useful biologic marker for hypoxia in uterine cervical cancer. PMID- 12095551 TI - Stage II endometrial carcinoma: prognostic factors and risk classification in 170 patients. AB - PURPOSE: Factors affecting outcome in patients with surgicopathologic Stage II endometrial cancer are poorly defined. The purpose of this study was to determine prognostic factors in a series of patients treated according to standardized protocols at a single institution. METHODS AND MATERIALS: One hundred and seventy patients referred to Princess Margaret Hospital after hysterectomy between 1984 and 1995 were retrospectively reviewed. One hundred and twenty patients received postoperative external beam radiotherapy and brachytherapy, 18 received external beam radiotherapy alone, five received brachytherapy alone, and 27 had no radiotherapy. RESULTS: With a median follow-up of 5.1 years, overall and disease free survival (DFS) at 5 years was 77% and 68%, respectively, and 24% of patients had relapsed. Significant independent adverse factors for DFS included age >65 (p = 0.0001), FIGO Stage IIB (p = 0.02), and capillary-lymphatic space (CLS) involvement (p = 0.0007). Prognostic factors for relapse were age (p = 0.0008), histologic grade (p = 0.01), and CLS (p = 0.01). A prognostic model based on the number of adverse prognostic factors (0-3) revealed that the 5-year survival rates for the four groups were as follows: 0%-85%, 1%-77%, 2%-55%, and 3%-11%. Combining the groups with 0 or 1 adverse factors resulted in a three-group variable that was strongly related to DFS (p < 0.0001). CONCLUSIONS: Patient age, stage, and CLS were significant factors for DFS, and age, grade, and CLS predicted time to relapse in Stage II endometrial cancer. A prognostic model for DFS using these factors can provide clinically meaningful outcome predictions. PMID- 12095552 TI - Breast-conserving therapy for ductal carcinoma in situ of the breast: the French Cancer Centers' experience. AB - PURPOSE: To assess the long-term outcome for women with ductal carcinoma in situ of the breast treated in current clinical practice by conservative surgery with or without definitive breast irradiation. METHODS AND MATERIALS: We analyzed 705 cases of ductal carcinoma in situ treated between 1985 and 1995 in nine French regional cancer centers; 515 underwent conservative surgery and radiotherapy (CS+RT) and 190 CS alone. The median follow-up was 7 years. RESULTS: The 7-year crude local recurrence (LR) rate was 12.6% (95% confidence interval [CI] 9.4 15.8) and 32.4% (95% CI 25-39.7) for the CS+RT and CS groups, respectively (p <0.0001). The respective 10-year results were 18.2% (95% CI 13.3-23) and 43.8% (95% CI 30-57.7). A total of 125 LRs occurred, 66 and 59 in the CS+RT and CS groups, respectively. Invasive or microinvasive LRs occurred in 60.6% and 52% of the cases in the same respective groups. The median time to LR development was 55 and 41 months. Nine (1.7%) and 6 (3.1%) nodal recurrences occurred in the CS+RT and CS groups, respectively. Distant metastases occurred in 1.4% and 3% of the respective groups. Patient age and excision quality (final margin status) were both significantly associated with LR risk in the CS+RT group: the LR rate was 29%, 13%, and 8% among women aged < or =40, 41-60, and > or =61 years (p <0.001). Even in the case of complete excision, we observed a 24% rate of LR (6 of 25) in women <40 years. Patients with negative, positive, or uncertain margins had a 7 year crude LR rate of 9.7%, 25.2%, and 12.2%, respectively (p = 0.008). RT reduced the LR rate in all subgroups, especially in those with comedocarcinoma (17% vs. 59% in the CS+RT and CS groups, respectively, p <0.0001) and mixed cribriform/papillary tumors (9% vs. 31%, p <0.0001). In the multivariate Cox regression model, young age and positive margins remained significant in the CS+RT group (p = 0.00012 and p = 0.016). Finally, the relative LR risk in the CS+RT group compared with the CS group was 0.35 (95% CI 0.25-0.51, p = 0.0001). Subsequent contralateral breast cancer occurred in 7.1% and 7.5% of the patients in the CS+RT and CS groups, respectively. CONCLUSION: Despite the absence of randomization, our results are extremely consistent with the updated National Surgical Adjuvant Breast Project B17 and European Organization for Research and Treatment of Cancer 10853 trials. We also noted that the LR risk was very high in women <40 years and/or in the case of incomplete excision. PMID- 12095553 TI - Pathologic tumor size and lymph node status predict for different rates of locoregional recurrence after mastectomy for breast cancer patients treated with neoadjuvant versus adjuvant chemotherapy. AB - PURPOSE: To compare the pathologic factors associated with postmastectomy locoregional recurrence (LRR) in breast cancer patients not receiving radiation who were treated with neoadjuvant chemotherapy (NEO) vs. adjuvant chemotherapy (ADJ). METHODS AND MATERIALS: We retrospectively analyzed the rates of LRR of subsets of women treated in prospective trials who underwent mastectomy and received chemotherapy but not radiation. These trials were designed to answer chemotherapy questions. There were 150 patients in the NEO group and 1031 patients in the ADJ group. In the NEO group, 55% had clinical Stage IIIA or higher vs. 9% in the ADJ group (p <0.001, chi-square test). RESULTS: Despite the more advanced clinical stage in the NEO group, the pathologic size of the primary tumor and the number of positive lymph nodes (+LNs) were significantly less in the NEO group than in the ADJ group (p <0.001 for both comparisons). However, the 5-year actuarial LRR rate was 27% for the NEO group vs. 15% for the ADJ group (p = 0.001, log-rank). The 5-year risk for LRR was higher in the NEO patients for all pathologic tumor sizes: 0-2 cm (18% vs. 8%, p = 0.011), 2.1-5 cm (36% vs. 15%, p <0.001), and >5 cm (46% vs. 28%, p = 0.028). The risk of LRR by the number of +LNs was similar in the NEO and ADJ groups, except for the subset of patients with > or =4 +LNs (53% vs. 23%, p <0.001). The rates of LRR in the patients with primary tumors measuring < or =2.0 cm and 1-3 +LNs were similar in both groups. However, for the patients with a pathologic tumor size of 2.1-5.0 cm and 1-3 +LNs, the LRR was higher in the NEO group than in the ADJ group (30% vs. 15%, p = 0.016). Most failures in this NEO subgroup had clinical Stage III disease. In a subset of NEO and ADJ patients matched for clinical stage, no significant differences were found in the rates of LRR according to primary tumor size and number of +LNs when these variables were analyzed independently. Again, however, differences were found in the subgroup of patients with tumors pathologically measuring 2.1-5.0 cm with 1-3 +LNs (32% NEO vs. 8% ADJ, p = 0.030). CONCLUSION: The rates of postmastectomy LRR for any pathologic tumor size are higher for patients treated with initial chemotherapy than for patients treated with initial surgery. Radiotherapy should be offered to all patients with > or =4 +LNs, tumor size >5 cm, or clinical Stage IIIA or greater disease, regardless of whether they receive neoadjuvant or postoperative chemotherapy. The information assessing LRR rates in patients with clinical Stage II disease who receive neoadjuvant chemotherapy, particularly if 1-3 lymph nodes remain pathologically involved, is insufficient to determine whether these patients should receive radiotherapy. PMID- 12095554 TI - Preliminary results of a phase I/II study of HDR brachytherapy alone for T1/T2 breast cancer. AB - PURPOSE: To investigate the feasibility, toxicity, cosmetic outcome, and local control of high-dose-rate (HDR) brachytherapy alone without whole breast external beam irradiation for early-stage breast carcinoma. METHODS AND MATERIALS: Between June 1997 and August 1999, 32 women diagnosed with a total of 33 AJCC Stage I/II breast carcinomas underwent surgical breast excision and postoperative irradiation using HDR brachytherapy interstitial implantation as part of a multi institutional clinical Phase I/II protocol. Eligible patients included those with T1, T2, N0, N1 (< or =3 nodes positive), and M0 tumors of nonlobular histologic features with negative surgical margins, no extracapsular lymph node extension, and a negative postexcision mammogram. Brachytherapy catheters were placed at the initial excision, reexcision, or either sentinel or full-axillary sampling. Direct visualization, surgical clips, and ultrasound and/or CT scan assisted in the delineation of the target volume, defined as the excision cavity plus a 2-cm margin. High-activity 192Ir (3-10 Ci) was used to deliver 340 cGy/fraction, 2 fractions/d, for 5 consecutive days, to a total dose of 34 Gy to the target volume. Source position and dwell times were calculated using standard volume optimization techniques. RESULTS: The median follow-up of all patients was 33 months, and the mean patient age was 63 years. The mean tumor size was 1.3 cm, and 55% had an extensive intraductal component. Three patients had positive axillary nodes. Two patients experienced moderate perioperative pain that required narcotic analgesics. No peri- or postoperative infections occurred. No wound healing problems and no significant skin reactions related to the implant developed. The Radiation Therapy Oncology Group late radiation morbidity scoring scheme was applied to the entire 33-case cohort. In the assessment of the skin, 30 cases were Grade 0-1 and 3 cases were Grade 2. Subcutaneous toxicity was scored as 11 patients with Grade 0, 3 with Grade 1, 8 with Grade 2, 3 with Grade 3, and 8 with Grade 4. Clinically evident fat necrosis occurred in 8 patients at a median of 7.5 months after HDR brachytherapy completion. The only variables significantly associated with Grade 3-4 toxicity were the number of source dwell positions and the volume of tissue encompassed by the prescription isodose shell. The global cosmetic scores after a minimum of 18 months' follow-up were 0 cases with poor, 4 with fair, 5 with good, and 24 with excellent scores. One case of ipsilateral breast tumor recurrence was diagnosed 23 months after HDR brachytherapy, for a 4-year actuarial recurrence rate of 3%. This failure appeared to be a new primary tumor, because it was histologically distinct from the initial tumor and was located 9 cm from the initial tumor bed and 3 cm from the edge of the implant volume. CONCLUSION: Radiotherapy of the tumor bed alone with HDR interstitial brachytherapy is associated with a 33% incidence of Grade 3 4 s.c. toxicity, but with generally favorable overall cosmetic results. The risk of toxicity appears to be primarily related to the implant volume. With limited follow-up, the incidence of ipsilateral breast tumor recurrence was low. PMID- 12095555 TI - Using PSA, biopsy Gleason score, clinical stage, and the percentage of positive biopsies to identify optimal candidates for prostate-only radiation therapy. AB - PURPOSE: An identification of prostate cancer patients most likely to benefit from prostate-only radiation was made based upon the pretreatment prostate specific antigen (PSA), biopsy Gleason score, clinical stage, percentage of positive biopsies, and the 5-year postoperative PSA outcome. METHODS: Between 1989 and 2000, 2099 patients underwent radical prostatectomy for clinically localized prostate cancer. The primary end points were pathologic evidence of seminal vesicle invasion 2(SVI), extracapsular extension (ECE) with or without positive surgical margins, and the 5-year postoperative PSA outcome. RESULTS: Pretreatment PSA, biopsy Gleason score, and clinical stage were used to assign patients to low-, intermediate-, and high-risk groups. These risk groups were stratified by the percentage of positive biopsies and the primary pathologic and biochemical outcomes examined. The rates of SVI, ECE with positive margin, and no biochemical evidence of disease (bNED) for low-risk patients with < or =50% positive biopsies were 2%, 7%, and 93%, respectively. Patients with >50% positive biopsies had higher rates of SVI and ECE (5% and 11%, respectively) and 52% bNED (p < 0.0001). For intermediate-risk patients with < or =17% positive biopsies, the rates of SVI, ECE with positive margin, and bNED were 3%, 9%, and 90%, respectively. As the percentage of positive biopsies increased above 17% in intermediate-risk patients, there was a statistically significant increase in SVI and ECE and a significant decrease in bNED. CONCLUSIONS: Low-risk patients with < or =50% positive biopsies and intermediate-risk patients with < or =17% positive biopsies had a very low risk of SVI and ECE with positive surgical margins. Given that the presence of SVI and ECE with positive surgical margins was uncommon (<10%) with a > or =90% PSA failure-free survival after radical prostatectomy, these patients may be optimal candidates for radiation therapy directed at the prostate only (prostate gland + 1.5-cm margin). PMID- 12095556 TI - Preliminary observations on biochemical relapse-free survival rates after short course intensity-modulated radiotherapy (70 Gy at 2.5 Gy/fraction) for localized prostate cancer. AB - PURPOSE: To compare the preliminary biochemical relapse-free survival rates between short-course intensity-modulated radiotherapy (SCIM-RT) delivering 70 Gy in 28 fractions and three-dimensional conformal radiotherapy (3D-CRT) delivering 78 Gy in 39 fractions. METHODS AND MATERIALS: Between January 1998 and December 1999, 166 patients were treated with SCIM-RT and 116 with 3D-CRT. The SCIM-RT cases were treated to 70 Gy (2.5 Gy/fraction) using 5 intensity-modulated fields using a dynamic multileaf collimator. The BAT transabdominal ultrasound system was used for localization of the prostate gland in all SCIM-RT cases. The 116 3D CRT cases were treated to 78.0 Gy (2.0 Gy/fraction). The study sample therefore comprised 282 cases; 70 Gy in 28 fractions is equivalent to 78 Gy in 39 fractions for late-reacting tissues, according to the linear-quadratic model. The median follow-up for all cases was 25 months (range 3-42). The median follow-up was 21 months for the SCIM-RT cases (range 3-31) and 32 months for the 3D-CRT cases (range 3-42). The follow-up period was shorter for the SCIM-RT cases, because SCIM-RT was started only in October 1998. Biochemical relapse was defined as 3 consecutive rising prostate-specific antigen levels after reaching a nadir. The analysis was then repeated with a more stringent definition of biochemical control: reaching and maintaining a prostate-specific antigen level of < or =0.5 ng/mL. Radiation Therapy Oncology Group toxicity scores were used to assess complications. RESULTS: For the 282 patients, the biochemical relapse-free survival rate at 30 months was 91% (95% confidence interval 88-95%). The biochemical relapse-free survival rate at 30 months for 3D-CRT vs. SCIM-RT was 88% (95% confidence interval 82-94%) vs. 94% (95% confidence interval 91-98%), respectively. The difference was not statistically significant between the two treatment arms (p = 0.084). The multivariate time-to-failure analysis using the Cox proportional hazards model for clinical parameters showed the pretreatment prostate-specific antigen level (p <0.001) and biopsy Gleason score (p <0.001) to be the only independent predictors of biochemical relapse. Clinical T stage (p = 0.66), age (p = 0.15), race (p = 0.25), and neoadjuvant androgen deprivation (p = 0.66) were not independent predictors of biochemical failure. SCIM-RT showed only a trend toward a better outcome on multivariate analysis (p = 0.058). Late rectal toxicity was limited; the actuarial combined Grade 2 and 3 late rectal toxicity rate at 30 months was 5% for SCIM-RT vs. 12% for 3D-CRT (p = 0.24). Grade 3 late rectal toxicity (rectal bleeding requiring cauterization) occurred in a total of 10 patients. The actuarial Grade 3 late rectal toxicity rate at 30 months was 2% for the SCIM-RT cases and 8% for the 3D-CRT cases (p = 0.059). Late urinary toxicity was rare in both groups. CONCLUSION: With the currently available follow up period (< or =30 months), the hypofractionated intensity-modulated radiotherapy schedule of 70.0 Gy delivered at 2.5 Gy/fraction had a comparable biochemical relapse profile with the prior 3D-CRT schedule delivering 78.0 at 2.0 Gy/fraction. The late rectal toxicity profile has been extremely favorable. If longer follow-up confirms the favorable biochemical failure and low late toxicity rates, SCIM-RT will be an alternative and more convenient way of providing dose escalation in the treatment of localized prostate cancer. PMID- 12095557 TI - Association of rectal toxicity with thermal dose parameters in treatment of locally advanced prostate cancer with radiation and hyperthermia. AB - PURPOSE: Although hyperthermia has been used for more than two decades in the treatment of pelvic tumors, little is known about the potential impact of heat on rectal toxicity when combined with other treatment modalities. Because rectal toxicity is a concern with radiation and may be exacerbated by hyperthermia, definition of the association of thermal dose parameters with rectal toxicity is important. In this report, we correlate rectal toxicity with thermal dose parameters for patients treated with hyperthermia and radiation for prostate cancer. METHODS AND MATERIALS: Thirty patients with T2b-T3b disease (1992 American Joint Committee On Cancer criteria) enrolled in a Phase II study of external beam radiation +/- androgen-suppressive therapy with two transrectal ultrasound hyperthermia treatments were assessed for rectal toxicity. Prostatic and anterior rectal wall temperatures were monitored for all treatments. Rectal wall temperatures were limited to 40 degrees C in 19 patients, 41 degrees C in 3 patients, and 42 degrees C in 8 patients. Logistic regression was used to estimate the log hazard of developing National Cancer Institute Common Toxicity Criteria Grade 2 toxicity based on temperature parameters. The following were calculated: hazard ratios, 95% confidence intervals, p values for statistical significance of each parameter, and proportion of variability explained for each parameter. RESULTS: Gastrointestinal toxicity was limited to Grade 2. The rate of acute Grade 2 proctitis was greater for patients with an allowable rectal wall temperature of >40 degrees C. In this group, 7 of 11 patients experienced acute Grade 2 proctitis, as opposed to 3 of 19 patients in the group with rectal wall temperatures limited to 40 degrees C (p = 0.004). Preliminary assessment of long term toxicity revealed no differences in toxicity. Hazard ratios for acute Grade 2 proctitis for allowable rectal wall temperature, average rectal wall Tmax, and average prostate Tmax were 9.33 (p = 0.01), 3.66 (p = 0.03), and 2.29 (p = 0.08), respectively. A model combining these three parameters explained 48.6% of the variability among groups. CONCLUSION: Rectal toxicity correlates with maximum allowable rectal wall temperature, average rectal wall Tmax, and average prostate Tmax for patients undergoing transrectal ultrasound hyperthermia combined with radiation for treatment of advanced clinically localized prostate cancer. Further definition of this association of thermal dose parameters with rectal toxicity in treatment of pelvic malignancies with hyperthermia should advance the goal of delivering thermal therapy in an effective yet safe manner. PMID- 12095558 TI - Clinical significance of increased gelatinolytic activity in the rectal mucosa during external beam radiation therapy of prostate cancer. AB - PURPOSE: Rectal toxicity (proctitis) is a dose-limiting factor in pelvic radiation therapy. Mucosal atrophy, i.e., net extracellular matrix degradation, is a prominent feature of radiation proctitis, but the underlying mechanisms are not known. We prospectively examined changes in matrix metalloproteinase (MMP)-2 and MMP-9 (gelatinase A and B) in the rectal mucosa during radiation therapy of prostate cancer, as well as the relationships of these changes with symptomatic, structural, and cellular evidence of radiation proctitis. METHODS AND MATERIALS: Seventeen patients scheduled for external beam radiation therapy for prostate cancer were prospectively enrolled. Symptoms of gastrointestinal toxicity were recorded, and endoscopy with biopsy of the rectal mucosa was performed before radiation therapy, as well as 2 and 6 weeks into the treatment course. Radiation proctitis was assessed by endoscopic scoring, quantitative histology, and quantitative immunohistochemistry. MMP-2 and MMP-9 were localized immunohistochemically, and activities were determined by gelatin zymography. RESULTS: Symptoms, endoscopic scores, histologic injury, and mucosal macrophages and neutrophils increased from baseline to 2 weeks. Symptoms increased further from 2 weeks to 6 weeks, whereas endoscopic and cellular evidence of proctitis did not. Compared to pretreatment values, there was increased total gelatinolytic activity of MMP-2 and MMP-9 at 2 weeks (p = 0.02 and p = 0.004, respectively) and 6 weeks (p = 0.006 and p = 0.001, respectively). Active MMP-2 was increased at both time points (p = 0.0001 and p = 0.002). Increased MMP-9 and MMP-2 at 6 weeks was associated with radiation-induced diarrhea (p = 0.007 and p = 0.02, respectively) and with mucosal neutrophil infiltration (rho = 0.62). CONCLUSIONS: Pelvic radiation therapy causes increased MMP-2 and MMP-9 activity in the rectal mucosa. These changes correlate with radiation-induced diarrhea and granulocyte infiltration and may contribute to abnormal connective tissue remodeling in radiation proctitis. PMID- 12095559 TI - Penile bulb imaging. AB - PURPOSE: Because of the apparent relationship between potency loss and radiation doses to the erectile bodies, there is increasing rationale for incorporating penile bulb dosimetry into treatment planning and posttreatment evaluation. Because the location and shape of the penile bulb have not been described in detail on various imaging modalities, we herein describe the anatomic boundaries of the penile bulb on computed tomography (CT), magnetic resonance imaging (MR), and transrectal ultrasound (TRUS), before and after brachytherapy. METHODS AND MATERIALS: Nonenhanced axial CT images were taken on a CTi CT Scanner (General Electric Medical Systems, Milwaukee, WI) with the patient in the supine position. Settings were at 300 ma, 140 kvp, 4-s scan time per slice, and collimation of 3 mm with data obtained at 3-mm intervals. Nonenhanced MR images were obtained with a 1.5 Tesla Signa Horizon LX Scanner using fast spin-echo T1-weighted (TR/TE, 466/20) and T2-weighted (TR/TE, 8000/90) images, with a slice thickness of 2 mm and an interslice gap of 0.5 mm. TRUS images were obtained with a Siemens SONOLINE Prima ultrasound machine at 6.0 MHz and a Winston-Barzell stepper unit. RESULTS: The penile bulb is best visualized on T2-weighted MR images in the axial, sagittal, and coronal planes, appearing as an oval-shaped, hyperintense midline structure. On axial CT imaging, the bulb of the penis is typically readily identifiable, bounded by the paired crura laterally, the corpora spongiosum anteriorly, and the levator ani posteriorly. The penile bulb is typically well visualized on transverse TRUS, but usually only faintly seen on sagittal TRUS. The bulb is partially obscured on postimplant CT and MR images, presumably because of implant-related edema. Bulb volumes vary markedly from patient to patient, ranging from 5.6 to 12.4 cc (median: 8.1 cc). CONCLUSION: Closer attention to penile erectile tissue doses should lead to improved external beam radiation and brachytherapy delivery. It will benefit the radiation oncology community to become familiar with these imaging findings, so that penile bulb dosimetry can be incorporated into our daily practice. PMID- 12095560 TI - The use of preferences to measure the benefit of adjuvant radiation therapy for stage I seminoma. AB - PURPOSE: In Stage I seminoma, treatment with radiation therapy (RT) after radical inguinal orchiectomy reduces the likelihood of relapse by 15%, but does not improve survival, thus making quality of life an important outcome measure. The purpose of this study was to use utilities to assess the quality of life benefits associated with adjuvant RT in this setting. MATERIALS AND METHODS: One hundred healthy men were interviewed using a utility assessment tool. Utilities for five health states were measured using the standard gamble technique: (A) adjuvant RT with 5% recurrence risk; (B) recurrence after RT, salvaged with chemotherapy; (C) orchiectomy alone with 20% recurrence risk; (D) recurrence after orchiectomy alone, salvaged with RT; and (E) recurrence after orchiectomy alone, salvaged with chemotherapy. RESULTS: The median age was 25. Utilities were highest for nonrecurrent health states, and lowest for recurrence salvaged with chemotherapy. All differences in utilities between health states were significant, except between states A and C and B and E. Variability in utilities was not explained by the sociodemographic factors examined. CONCLUSIONS: Our results suggest that healthy males do not value the 15% reduction in recurrence risk achievable with adjuvant RT. However, they do predict that an actual recurrence, especially one requiring salvage chemotherapy, will lead to significant decline in quality of life. We intend to use these utilities to further evaluate the cost-effectiveness of RT in this setting. PMID- 12095561 TI - Phase II study of tolerance and efficacy of hyperfractionated radiotherapy and 5 fluorouracil, cisplatin, and paclitaxel (Taxol) in stage III and IV inoperable and/or unresectable head-and-neck squamous cell carcinoma: A-2 protocol. AB - PURPOSE: To determine the toxicity and efficacy of concurrent 5-fluorouracil (5 FU), cisplatin, and paclitaxel (Taxol) and hyperfractionated radiotherapy in locally advanced squamous cell carcinoma of the head and neck. METHODS AND MATERIALS: Twenty-seven patients were entered into this Phase II trial. Eligible patients had Stage III or IV head-and-neck squamous cell carcinoma arising from the oral cavity, hypopharynx, oropharynx, nasopharynx, or larynx. The plan of treatment consisted of hyperfractionated radiotherapy (74.4 Gy at twice daily fractions of 1.2 Gy). Chemotherapy was given on Weeks 1, 5, and 8 as follows: 5 FU at 750 mg/m2 as a constant infusion for 24 h for 3 days; cisplatin at 50 mg/m2 in 250-500 mL D5 0.5 NS or NS infusion during 2-4 h, and paclitaxel at 70 mg/m2 infused in 500 mL NS during 3 h. RESULTS: The overall survival rate of the entire group was 81.5%, 66.7%, and 63% at 1, 2, and 3 years, respectively. The median follow-up was 40.2 months (range 30-62). Of the 27 patients, 19 (70%) had a complete response and an overall survival rate of 100% at 1 year and 94% at 2 and 3 years. The disease-free survival rate of the latter group was 95% at 1 year and 84% at 2 and 3 years. Of the 27 patients, 18 (67%) maintained the complete response until the last follow-up visit or death. Percutaneous endoscopic gastrostomy dependency occurred for a median of 7.1 months. Grade 3 and 4 mucositis occurred in 20 and 3 patients, respectively. Six patients were hospitalized for leukopenic fever. Late toxicities included L'Hermitte syndrome (n = 3), osteoradionecrosis (n =1), hypothyroidism (n = 4), paresthesias (n = 1), aspiration pneumonia (n = 3), and esophageal strictures (8 patients underwent dilation). CONCLUSION: Combining hyperfractionated radiotherapy concurrently with 5-FU, cisplatin, and paclitaxel results in acceptable efficacy and toxicity. However, although a locoregional control benefit is suggested by the preliminary results of this trial, it needs to be confirmed in a prospective randomized trial. PMID- 12095562 TI - Supraglottic carcinoma: patterns of failure and salvage treatment after curatively intended radiotherapy in 410 consecutive patients. AB - PURPOSE: In a series of consecutive patients with squamous cell carcinoma of the supraglottic larynx, in which almost all were treated by primary radiotherapy, the study describes the path from diagnosis to cure or death, and evaluates the patterns of failure and the treatment of recurrences. METHODS AND MATERIALS: The analysis included 410 patients, 104 females and 306 males, treated between 1963 and 1991. Most patients were in Stage I (33%), and the remaining were in Stage II (18%), III (23%), and IV (26%). Primary intended curative treatment was delivered in 398 (radiotherapy, 394; surgery, 4) of 410 cases (98%). RESULTS: Initial radical treatment resulted in 173 cured patients and 225 patients with a recurrence. Curatively intended salvage could be applied in 158 patients: surgery in 154 patients (74 cured) and radiotherapy in 4 (none cured). Overall, 247 patients (60%) obtained tumor control, 179 (44%) without a laryngectomy. Sixty three patients had a total laryngectomy, and five had a partial laryngectomy. The 5-year locoregional tumor control, disease-specific survival, and overall survival rates were 43%, 61%, and 47%, respectively. With a follow-up of 20 years posttreatment, 91 new primary malignant tumors were detected. CONCLUSIONS: Radiotherapy is effective in the treatment of supraglottic laryngeal carcinoma, and the patients have a relatively good prognosis. Many patients retained their larynx intact. Recurrence after primary radiotherapy can be treated by surgery, with a high success rate even in advanced stages. Development of second primary cancer is a growing problem. PMID- 12095563 TI - Health-related quality of life for nasopharyngeal carcinoma patients with cancer free survival after treatment. AB - PURPOSE: To evaluate the health-related quality of life (HR-QOL) of nasopharyngeal cancer (NPC) patients with cancer-free survival after treatment and to investigate the factors correlated with their HR-QOL. METHODS AND MATERIALS: One hundred eighty-two NPC patients with cancer-free survival of more than 2 years after treatment were enrolled in the study. Data from the same number of people without a history of cancer, who had come to the hospital for health checkups, were also collected for comparison. The Chinese SF-36 questionnaire and nine items about head-and-neck functional impairments (HNFI) were self-reported by all participants at the clinics. Data relating to sociodemographic factors, cancer stage, and treatment of NPC survivors were analyzed. RESULTS: Psychometric tests revealed the excellent internal reliability (Cronbach's alpha: 0.87-0.96) and discriminative validity of the Chinese SF-36 used in Taiwan. Most functional domains of the Chinese SF-36 and all nine HNFI items were significantly worse in NPC survivors than in control subjects. No cancer or treatment-related variables significantly correlated with any functional domains of SF-36 or any items of HNFI for NPC survivors. Economic status, educational level, occupational status, and the number of comorbidities were the variables that significantly correlated with most functional domains of SF-36 for NPC survivors. Patients with more sufficient economic status, higher educational levels, with employment, or without comorbidity tended to enjoy better HR-QOL as detected by the SF-36. Salivation, hearing, and swallowing dysfunctions were the top three HNFI that disturbed NPC survivors. Economic status remained the most significant variable correlated with HNFI, including salivation, swallowing, neck stiffness, taste, and phonation. Survivors with better economic status reported less severe HNFI. CONCLUSIONS: NPC survivors had worse HR-QOL than healthy control subjects in the study. Socioeconomic status was the most significant variable that correlated with the HR-QOL of NPC survivors. This result might indicate that patients' superior individual characteristics and financial resources are important variables determining their ability to cope with cancer and treatment complications affecting their HR-QOL. PMID- 12095564 TI - Limitations of conventional doses of chemoradiation for unresectable biliary cancer. AB - PURPOSE: To determine, in a retrospective review, the limitations of definitive chemoradiation in the treatment of patients with unresectable extrahepatic cholangiocarcinoma and generate testable hypotheses for future prospective clinical trials. METHODS AND MATERIALS: Between 1957 and 2000, 52 patients with localized, unresectable cholangiocarcinoma were treated with radiotherapy (RT) with or without concurrent chemotherapy. Unresectable disease was defined, by evidence on imaging studies or at surgical exploration, as localized tumor abutting or involving the main portal vein, tumor involvement of secondary biliary radicals, or evidence of nodal metastases. Patients were grouped according to the RT dose: 27 patients received a total dose of 30 Gy (Group 1), 14 patients received 36-50.4 Gy (Group 2), and 11 patients received 54-85 Gy (Group 3). 192Ir intracavitary boosts (median 20 Gy) were delivered in 3 patients, and an intraoperative boost (20 Gy) was used in 1 patient. Of the 52 patients, 38 (73%) received concomitant protracted venous infusion of 5 fluorouracil (200-300 mg/m2 daily, Monday through Friday). Kaplan-Meier analysis was used to calculate the actuarial 1-year and median overall survival (OS), radiographic local progression, symptomatic progression, and distant failure. Treatment-related variables and prognostic factors were evaluated using the log rank test. RESULTS: The first site of disease progression was local in 72% of cases. The actuarial local progression rate at 12 months for all patients was 59%. The median time to radiographic local progression was 9, 11, and 15 months in Groups 1, 2, and 3, respectively (p = 0.48). Fifteen percent of all patients developed metastatic disease (1-year OS rate 18%). The median survival rate for all patients was 10 months (1-year OS rate 44%). The RT dose, use of concurrent chemotherapy, histologic grade, initial extent of liver involvement, and extent of vascular involvement had no influence on radiographic local progression or OS. Grade 3 or greater toxicity was similar in all dose groups (22% vs. 14% vs. 27%, p = 0.718). CONCLUSION: The primary limitation of definitive chemoradiation was local progression. Although the small patient numbers limited the statistical power of this study, a suggestion of improved local control was found with the use of higher RT doses. To address this pattern of failure, future prospective investigation using high-dose conformal RT with novel cytotoxic and/or biologic agents with radiosensitizing properties is warranted. PMID- 12095565 TI - Final results of a prospective study of the prognostic value of the time to develop motor deficits before irradiation in metastatic spinal cord compression. AB - PURPOSE: Metastatic spinal cord compression often requires urgent treatment selection, which could be facilitated by strong prognostic factors. Because only the type of primary tumor and pretreatment ambulatory status are known as prognostic factors for functional outcome, we investigated the prognostic value of the time of motor deficit development before radiotherapy (RT). METHODS AND MATERIALS: Ninety-eight patients were included between November 1998 and April 2000. Three subgroups were formed, according to time of motor deficit development before RT: 1-7 days (n = 31), 8-14 days (n = 31), and >14 days (n = 36). Ambulatory rates and motor function were evaluated for < or =24 weeks after RT. In a multivariate analysis, all three prognostic factors and radiation parameters were included. RESULTS: In the >14-day subgroup, improvement occurred significantly (p < 0.001) more often than in the other subgroups (86% vs. 29% and 10%) and the posttreatment ambulatory rate was significantly higher (86% vs. 55% and 35%, p = 0.026). Multivariate analysis revealed the time of development of motor deficits before RT to be the strongest prognostic factor. CONCLUSION: Functional outcome is significantly better with slower development of motor deficits before RT. This new, independent, prognostic factor must be considered in future trials aiming to define an optimal RT schedule. PMID- 12095566 TI - Phase II study of topotecan plus cranial radiation for glioblastoma multiforme: results of Radiation Therapy Oncology Group 9513. AB - PURPOSE: A Phase II trial was conducted by the Radiation Therapy Oncology Group (RTOG) to compare the survival of patients with glioblastoma multiforme treated with topotecan combined with standard cranial radiotherapy (RT) for matched patients treated in prior RTOG studies. A secondary objective was to document the acute and late toxicities of this combination of chemotherapy and RT. METHODS AND MATERIALS: Eighty-seven patients with histologically confirmed glioblastoma multiforme received standard cranial RT (60 Gy/30 fractions in 6 weeks) plus topotecan 1.5 mg/m2 per day i.v. for 5 d/wk every 3 weeks for 3 cycles. Eighty four patients were evaluated, of whom 60 (71%) were > or =50 years, 44 (52%) were men, and 61 (73%) had a Karnofsky performance status of > or =80. Twenty-nine percent of patients had undergone biopsies, 48% partial resections, and 21% gross total resections. Two resections were unspecified as to the extent of tumor removal. Fourteen percent of patients were recursive partitioning analysis Class III, 46% were Class IV, 35% were Class V, and 5% were Class VI. RESULTS: The median survival was 9.3 months. Sixty-seven patients (80%) had progression. The 1 year survival rate was 32%. One patient remained alive without recurrence. RTOG 9513 patients were matched with patients in an RTOG clinical trial database from previous clinical trials. The matching variables were age, Karnofsky performance status, mental status, and prior surgery. No statistically significant difference was found between the survival of the study patients and that of the matched patients from the RTOG database. Fifty-four percent of patients had Grade IV acute toxicity. The toxicity was primarily hematologic. Four patients had Grade III late central nervous system toxicities. CONCLUSION: Topotecan administered at a dose of 1.5 mg/m2 per day i.v. for 5 d/wk every 3 weeks for 3 cycles given concurrently with standard cranial RT for glioblastoma does not produce a statistically significant survival advantage over previously tested therapies. Other methods of administration of topotecan or other camptothecins may provide more effective radiosensitization. PMID- 12095567 TI - Fractionated stereotactic radiotherapy for the treatment of vestibular schwannomas: combined experience of the Toronto-Sunnybrook Regional Cancer Centre and the Princess Margaret Hospital. AB - PURPOSE: To evaluate the efficacy and toxicity of fractionated stereotactic radiotherapy (FSRT) for vestibular schwannomas in patients treated at two university-affiliated hospitals. METHODS AND MATERIALS: Thirty-nine patients were treated between April 1996 and September 2000. The median age was 56 years (range: 29-80), and median maximal tumor diameter was 20 mm (range: 9-40). A total of 11 patients had fifth and/or seventh cranial nerve dysfunction before irradiation; 2 patients had only facial weakness, 5 patients had only facial numbness, and 4 patients had both facial weakness and numbness. Thirty-three patients were treated with primary FSRT, and 6 patients were treated for recurrent or persistent disease after previous surgery. All patients were treated with 6-MV photons using a stereotactic system with a relocatable frame. The 39 patients received 50 Gy in 25 fractions over 5 weeks. Median follow-up was 21.8 months (range: 4.4-49.6). RESULTS: Local control was achieved in 37 patients (95%). Two patients experienced deterioration of their symptoms at 3 and 20 months as a result of clinical progression in one case and tumor progression in the other and underwent surgery post FSRT. A total of 19/28 (67.9%) patients preserved serviceable hearing after FSRT. Deterioration of the facial and trigeminal nerves was observed in only 2 patients who were treated with surgery post FSRT. CONCLUSION: FSRT provided excellent tumor control with minimal morbidity and good hearing preservation in this cohort of patients. Longer follow up is required to confirm long-term control rates. PMID- 12095568 TI - The role of Gamma Knife radiosurgery in the management of cavernous sinus meningiomas. AB - PURPOSE: To evaluate the efficacy of Gamma Knife (GK) radiosurgery in terms of neurologic improvement and tumor growth control (TGC) in a large series of patients with cavernous sinus meningioma (CSM). METHODS AND MATERIALS: One hundred thirty-eight patients with CSM (28 males, 110 females; mean age: 56.2 years) were treated with GK between February 1993 and February 2001. GK was used as a first-choice treatment in 68/138 patients and as postoperative adjuvant therapy in 70/138. In 32 patients, it was possible to compare the size of the planned treatment volume to tumor volume using the conformity index (CI); optimal CI values were taken to be < or =1.5 (range: 0.94-2.24). RESULTS: A follow-up (FU) period of at least 12 months was available for 111 patients (median: 48.2 months, range: 12.1-84.5 months). Clinical conditions were improved or stable in 107/111 patients (96.5%). Neurologic recovery was observed in 76% of cases treated by GK alone and in 56.5% of adjuvant treatments (p < 0.03). Adequate TGC was documented in 108/111 tumors (97%), with shrinkage/disappearance in 70/111 (63%) and no variation in volume in 38/111 (34%); the overall actuarial progression-free survival rate at 5 years was 96%. Tumor size regression was observed in 79.5% of patients with FU >30 months, compared with 47.5% of patients with FU <30 months (p < 0.001). One hundred percent TGC was shown in treated patients with a CI < or =1.5 (20/32), compared with 92% TGC in cases with a CI >1.5 (p < 0.15, NS). Radiosurgical sequelae were transient in 4/111 cases (3.5%) and permanent in one case (1%). CONCLUSIONS: For the FU period of our series (median: >4 years), GK radiosurgery seems to be both safe (permanent morbidity 1%) and effective (96% neurologic improvement/stability, 97% overall TGC, 96% actuarial TGC at 5 years) and might be considered as a first-choice treatment for selected patients with CSM. PMID- 12095569 TI - An evaluation of the relationship between the quality of prophylactic cranial radiotherapy in childhood acute leukemia and institutional experience: a Quality Assurance Review Center-Pediatric Oncology Group study. AB - PURPOSE: The Pediatric Oncology Group Protocol 9404 was a prospective clinical trial of two forms of chemotherapy in childhood T-cell acute lymphoblastic leukemia and advanced stage T-cell lymphoblastic non-Hodgkin's lymphoma. The protocol called for prophylactic C1 whole brain external beam irradiation, 18 Gy in 2 Gy/fraction for 9 fractions. We hypothesized that a correlation would be found between the number of children irradiated on protocol by an institution and the compliance rate of that institution with radiotherapy quality assurance (QA) guidelines. We also hypothesized that QA compliance would improve as the study progressed. METHODS AND MATERIALS: We scored the radiation dose as a minor deviation from protocol guidelines if the dose to the prescription point differed from the protocol by 6-10%, and a major deviation if it differed from protocol by > 10%. Treatment volumes were scored as a minor deviation if the margins were less than specified or the fields were excessively large. A major deviation was defined as the transection of a potential leukemia-bearing volume such as would be caused by blocking the cribriform plate, optic nerve, or temporal lobe. When the treating physician submitted a treatment plan and simulator film at the initiation of therapy to the Quality Assurance Review Center (QARC), a rapid turn around review of the plan and suggestions for improvement was provided. At the end of therapy, all simulator and port films were reviewed at the QARC. RESULTS: We reviewed the data from 353 patients treated at 73 institutions in the United States, Canada, and Europe. Of these patients, 2% (n = 7) were not assessable for QA because of incomplete information. Minor quality deviations were found in 27.7% of patients (n = 98) and major deviations in 7.9% (n = 28). The frequency of major deviations for institutions placing 1-4 patients on study was 11% vs. 5.5% for institutions placing > or =5 patients (p < 0.09). The frequency of minor deviations was 28.6% for institutions placing 1-4 patients on study vs. 27.1% for institutions placing >5 patients (p not significant). The frequency of major deviations fell over time (1996-1997, 15.5% vs. 1998-2001, 4.7%, p < 0.001). The frequency of minor deviations did not (1996-1997, 29.9% vs. 1998-2001, 26.9%, p not significant). CONCLUSION: For a relatively simple radiotherapy field, with clearly written protocol guidelines and rapid turn-around corrections from QARC, the rate of minor deviations was no different between institutions placing 1-4 patients on study and those placing > or =5. A trend (p < 0.09) was noted, however, for major deviations to decrease as a function of institutional experience, as well as over time (p < 0.001), supporting the validity of the hypothesis that pediatric clinical experience matters in QA for C1 whole brain leukemia radiotherapy. PMID- 12095571 TI - Modeling volume effects of experimental brachytherapy in the rat rectum: uncovering the limitations of a radiobiologic concept. AB - PURPOSE: To analyze the significance of volume effects in experimental brachytherapy, based on modeling normal tissue complication probability. METHODS AND MATERIALS: Experimental brachytherapy in the rat rectum was based on an eight step 2.5-mm step size source configuration for 192Ir, afterloaded into an unshielded polystyrene applicator. Volume effects were studied using a half circumferential lead-shielded applicator and a shorter (two-step) source configuration. The main end point was rectal stenosis. RESULTS: Rectal stenosis was always caused by a radiation ulcer. With the shielded configuration, single dose ED50 (50% incidence of rectal stenosis) increased from 23 Gy to 36.5 Gy. Single-dose ED50 for the short configuration was 77.9 Gy. The data showed a reasonable fit to a three-parameter version of the biophysical model described by Jackson et al. (1995). This model assumes that organs consist of a large number of radiobiologically independent subunits and that radiation causes a complication if the fraction of the organ damaged is greater than its functional reserve. The fraction of the organ damaged is calculated summing over fractions of the organ damaged at each dose level. The calculated mean functional reserve (nu50) of the rat rectum, assuming a cumulative functional reserve distribution in the group of experimental rats, was 0.53. CONCLUSIONS: The volume effect observed within small brachytherapy volumes agreed well with clinical experience of large tolerance doses in contact X-ray therapy. However, the nu50 value was comparable to the high functional reserve value reported for liver. Experimental volume effects probably reflect repair processes originating in the areas adjacent to small radiation fields of brachytherapy more than the radiobiologic characteristics of the cells in the irradiated volume. PMID- 12095570 TI - Self-expandable covered metallic esophageal stent impregnated with beta-emitting radionuclide: an experimental study in canine esophagus. AB - PURPOSE: A specially designed self-expandable covered metallic stent impregnated with the beta-emitting radioisotope 166Ho (166Ho, energy: 1.85 and 1.76 MeV, T12: 26.8 h) was developed at our institute for the purpose of intraluminal palliative brachytherapy, as well as for treating malignant esophageal stricture and swallowing difficulty. The aim of this study was to evaluate the tissue response to brachytherapy and the safety of the radioactive metallic stent with regard to the normal canine esophagus before clinical application. METHODS AND MATERIALS: 166Ho was impregnated into the polyurethane membrane (50 micron thickness) covering the outer surface of a self-expandable metallic stent (diameter, 18 mm; length, 40 mm). Stents with radioactivity 4.0-7.8 mCi (Group A, n = 15), 1.0-1.8 mCi (Group B, n = 5), and 0.5-0.7 mCi (Group C, n = 5) were placed in the esophagi of 25 healthy beagle dogs, and the stents were tightly anchored surgically to prevent migration. The estimated radiation dose calculated by Monte Carlo simulation was 194-383 Gy in Group A, 48-90 Gy in Group B, and 23-32 Gy in Group C. The dogs were killed 8-12 weeks after insertion of the stents, and histologic examinations of the esophageal walls were performed. RESULTS: In Group A, 3 of 15 dogs died of wound infection, so specimens were obtained from only 12 dogs; all 12 cases showed esophageal stricture with mucosal ulceration. Microscopically, severe fibrosis and degeneration of the muscular propria were found in 3 dogs, complete fibrosis of the entire esophageal wall was found in 7 dogs, and esophageal fibrosis with radiation damage within periesophageal soft tissue was found in 2 dogs. However, esophageal perforation did not develop, despite extremely high radiation doses. In Group B, glandular atrophy and submucosal fibrosis were found, but the muscular layer was intact. In Group C, no histologic change was found in 3 dogs, but submucosal inflammation and glandular atrophy with intact mucosa were found in 2 dogs. CONCLUSIONS: A radioactive, self expandable covered metallic stent can be used as an alternative therapeutic modality for the palliative treatment of malignant esophageal stricture. PMID- 12095572 TI - Electron conformal radiotherapy using bolus and intensity modulation. AB - PURPOSE: Conformal electron beam therapy can be delivered using shaped bolus, which varies the penetration of the electrons across the incident beam so that the 90% isodose surface conforms to the distal surface of the planning target volume (PTV). Previous use of this modality has shown that the irregular proximal surface of the bolus causes the dose heterogeneity in the PTV to increase from 10%, the typical dose spread of a flat-water surface to approximately 20%. The present work evaluates the ability to restore dose homogeneity by varying the incident electron intensity. METHODS AND MATERIALS: Three patients, one each with chest wall, thorax, and head-and-neck cancer, were planned using electron conformal therapy with bolus, with and without intensity modulation. Resulting dose distributions and dose-volume histograms were compared with non-intensity modulated bolus plans. RESULTS: In all cases, the DeltaD(90%-10%) for the PTV was reduced; for example, for the head-and-neck case, the DeltaD(90%-10%) for the PTV was reduced from 14.9% to 9.2%. This reduction in dose spread is a direct result of intensity modulation. CONCLUSIONS: The results showed that intensity-modulated electron beams could significantly improve the dose homogeneity in the PTV for patients treated with electron conformal therapy using shaped bolus. PMID- 12095573 TI - Conformal radiotherapy optimization with micromultileaf collimators: comparison with radiosurgery techniques. AB - PURPOSE: Conformal radiotherapy (CRT) consists of irradiating the target volume while avoiding the healthy peripheral tissues and organs at risk as far as possible. One technique used to treat intracranial tumors consists of using micromultileaf collimators (MMLCs). Given the dose constraints involved, it is of interest to optimize MMLC irradiation parameters and compare the results of this technique with those of conventional radiosurgery (RT) techniques (Gamma Knife and linear accelerator stereotactic RT). METHODS AND MATERIALS: MMLC protocols are optimized in two stages. The orientation of the fields, delimited by a beam's eye view technique, is determined using a genetic algorithm method. The weighting of the fields and subfields when using intensity modulation and the position of the leaves are optimized using a simulated annealing method. We compared the results obtained for 8 clinical cases using 5 intensity-modulated fields with those obtained using the two radiosurgery techniques. The comparison indexes are those defined by the Radiation Therapy Oncology Group (RTOG). RESULTS: The results of this study demonstrated the advantages of using intensity modulation and the improvement obtained for the RTOG indexes in the case of CRT with MMLC, although the healthy peripheral tissues were less exposed to radiation with the radiosurgery techniques. The results also highlight the difficulty encountered with radiosurgery techniques in obtaining satisfactory dose homogeneity when the protocol is defined with numerous iosocenters. CONCLUSION: In CRT with MMLC, intensity modulation makes it possible to reduce the number of fields used. It is especially useful to optimize the orientations in the case of target volumes of complex shape or when volumes at risk are in the vicinity of the target. If used correctly, MMLC can be a valuable alternative to conventional radiosurgery techniques. PMID- 12095574 TI - Image fusion between 18FDG-PET and MRI/CT for radiotherapy planning of oropharyngeal and nasopharyngeal carcinomas. AB - PURPOSE: Accurate diagnosis of tumor extent is important in three-dimensional conformal radiotherapy. This study reports the use of image fusion between (18)F fluoro-2-deoxy-D-glucose positron emission tomography (18FDG-PET) and magnetic resonance imaging/computed tomography (MRI/CT) for better targets delineation in radiotherapy planning of head-and-neck cancers. METHODS AND MATERIALS: The subjects consisted of 12 patients with oropharyngeal carcinoma and 9 patients with nasopharyngeal carcinoma (NPC) who were treated with radical radiotherapy between July 1999 and February 2001. Image fusion between 18FDG-PET and MRI/CT was performed using an automatic multimodality image registration algorithm, which used the brain as an internal reference for registration. Gross tumor volume (GTV) was determined based on clinical examination and 18FDG uptake on the fusion images. Clinical target volume (CTV) was determined following the usual pattern of lymph node spread for each disease entity along with the clinical presentation of each patient. RESULTS: Except for 3 cases with superficial tumors, all the other primary tumors were detected by 18FDG-PET. The GTV volumes for primary tumors were not changed by image fusion in 19 cases (89%), increased by 49% in one NPC, and decreased by 45% in another NPC. Normal tissue sparing was more easily performed based on clearer GTV and CTV determination on the fusion images. In particular, parotid sparing became possible in 15 patients (71%) whose upper neck areas near the parotid glands were tumor-free by 18FDG-PET. Within a mean follow-up period of 18 months, no recurrence occurred in the areas defined as CTV, which was treated prophylactically, except for 1 patient who experienced nodal recurrence in the CTV and simultaneous primary site recurrence. CONCLUSION: This preliminary study showed that image fusion between 18FDG-PET and MRI/CT was useful in GTV and CTV determination in conformal RT, thus sparing normal tissues. PMID- 12095575 TI - Evaluation of deep inspiration breath-hold lung treatment plans with Monte Carlo dose calculation. AB - PURPOSE: To evaluate dosimetry of deep inspiration breath-hold (DIBH) relative to free breathing (FB) for three-dimensional conformal radiation therapy of lung cancer with 6-MV photons and Monte Carlo (MC) dose calculations. METHODS AND MATERIALS: Static three-dimensional conformal radiation therapy, 6-MV plans, based on DIBH and FB CT images for five non-small-cell lung cancer patients, were generated on a clinical treatment planning system with equivalent path length tissue inhomogeneity correction. Margins of gross to planning target volume were not reduced for DIBH plans. Cord and lung toxicity determined the maximum treatment dose for each plan. Dose distributions were recalculated for the same beams with an MC dose calculation algorithm and electron density distributions derived from the CT images. RESULTS: MC calculations showed decreased target coverage relative to treatment-planning system predictions. Lateral disequilibrium caused more degradation of target coverage for DIBH than for FB (approximately 4% worse than expected for FB vs. 8% for DIBH). However, with DIBH higher treatment doses could be delivered without violating normal tissue constraints, resulting in higher total doses to gross target volume and to >99% of planning target volume. CONCLUSIONS: If DIBH enables prescription dose increases exceeding 10%, MC calculations indicate that, despite lateral disequilibrium, higher doses will be delivered to medium-to-large, partly mediastinal gross target volumes, providing that 6-MV photons are used and margins are not reduced. PMID- 12095576 TI - On the dosimetric accuracy of a Sievert integration model in the proximity of 192Ir HDR sources. AB - PURPOSE: To investigate the efficacy of a Sievert integration model in dosimetry close to 192Ir high-dose-rate brachytherapy sources and validate its accuracy and potential to resolve dosimetric differences between these sources in the cm and mm distance ranges relevant to interstitial and intravascular brachytherapy applications, respectively. METHODS AND MATERIALS: The dosimetric quantities of the generalized Task Group 43 formalism, as well as dose rate profiles in polar and Cartesian coordinates, are calculated, and results are compared to corresponding Monte Carlo data in the literature. RESULTS: Sievert calculations were found in excellent agreement with corresponding Monte Carlo published results. Dose rate polar angle profiles in the cm distance range depended significantly on corresponding anisotropy function data, whereas in the mm distance range, dose rate polar angle profiles are governed by the corresponding geometry function profiles, because anisotropy proved insignificant. Radial dose functions of the sources were found comparable. A simple equation for the calculation of the dose rate constant of the sources within clinically acceptable accuracy is provided. CONCLUSIONS: The particular Sievert model proved capable of resolving dosimetric differences of the sources and provides results within clinical accuracy. Therefore, it constitutes a useful tool for dosimetry in clinical practice and especially in intravascular applications, where there is currently a lack of available dosimetric data. PMID- 12095577 TI - In regard to Purdy JA, Michalski JM. Does the evidence support the enthusiasm over 3D conformal radiation therapy and dose escalation in the treatment of prostate cancer? Int J Radiat Oncol Biol Phys 2001;51:867-870. PMID- 12095579 TI - In regard to Giraud et al.: Evaluation of microscopic tumor extension in non small-cell lung cancer for three-dimensional conformal radiotherapy planning. IJROBP 2000;48:1015-1024. PMID- 12095581 TI - In regard to Sherertz et al., IJROBP 2001; 51:1241-1245. PMID- 12095583 TI - In regard to Intensity-Modulated Radiotherapy Collaborative Working Group, IJROBP 2001; 51:880-914. PMID- 12095584 TI - Consensus statement on scientific data from clinical trials and investigators' responsibilities and rights. PMID- 12095586 TI - Standardized patient care guidelines reduce infectious morbidity in appendectomy patients. AB - BACKGROUND: Surgical wound infection and intra-abdominal abscess remain common infectious complications after appendectomy, especially in the setting of a perforated or gangrenous appendix. We therefore developed a clinical protocol for the management of appendicitis to decrease postoperative infectious complications. METHODS: Between January 1, 1999, and December 31, 1999, 206 patients with appendicitis were treated on protocol. Retrospectively, the charts were reviewed for all protocol patients as well as for 232 patients with appendicitis treated in the year prior to protocol initiation. Data were collected on surgical wound infections and intra-abdominal abscesses. RESULTS: There were significantly fewer infectious complications in the protocol group than in the nonprotocol group (20 [9%] versus 8 [4%]; P <0.05). In patients with a perforated or gangrenous appendix, the infectious complication rate was reduced from 33% to 13% (P <0.05). CONCLUSIONS: The incidence of infectious complications after appendectomy can be significantly reduced with a standardized approach to antibiotic therapy and wound management. PMID- 12095588 TI - Preoperative evaluation by magnetic resonance imaging in patients with bowel obstruction. AB - BACKGROUND: Bowel obstruction is a problematic condition because the main clinical issue is to determine whether emergency laparotomy or observation with a long tube is required. The recent development of imaging diagnostic modalities such as magnetic resonance imaging (MRI) is thought to be promising to support therapeutic decisions in patients with bowel obstruction. METHODS: Twenty-seven patients with bowel obstruction who underwent laparotomy were evaluated by plain x-ray film, computed tomography (CT) scan, and MRI preoperatively with regard to the presence or absence of bowel obstruction, and the site and cause of bowel obstruction. Diagnostic accuracies were compared among these radiological modalities. RESULTS: The presence of bowel obstruction was detected in 22 (81.5%) of 27 patients by plain abdominal x-ray film, in 24 (92.3%) of 26 patients by CT scan, and in 25 (92.6%) of 27 patients by MRI. The sites of obstruction were consistent with surgical findings in 25 (92.6%) of 27 patients by MRI, and in 15 (57.7%) of 26 patients by CT scan. The causes of bowel obstruction were accurately diagnosed by MRI in 25 (92.6%) of 27 patients, and in 23 (88.5%) of 26 patients by CT scan. CONCLUSIONS: MRI could identify the presence and the site and cause of bowel obstruction in most of the cases. MRI is assumed to be superior to CT scan in the preoperative diagnosis of bowel obstruction. PMID- 12095589 TI - The establishment of a surgical interest society for medical students. AB - With the advent of initiatives by many medical schools to attract students to generalist fields, the curriculum at these institutions has undergone substantial change. In many instances these changes include the abbreviation of exposure to specialty fields such as surgery. Consequently, the exposure of medical students to the surgical discipline and surgeons may be decreased at these institutions. These changes are particularly concerning in light of studies that suggest that these important interactions are the primary influences that lead students to pursue a career in specific fields. It is also interesting to note that these trends in decreasing exposure to surgical fields seem to correlate with recent increases in the number of unfilled categorical residency positions in general surgery. This article focuses on the experiences of a group of students and faculty mentors at the University of Virginia School of Medicine as they set about creating an extracurricular opportunity for students to explore interests in surgical fields. We shall present the thoughts and rationale we used in planning the establishment of our own student surgical interest society, as well as the manner in which we ultimately went about constructing this organization. It is our hope that this information will provide some ideas for the creation of similar societies at other institutions. PMID- 12095590 TI - The timing of fracture treatment in polytrauma patients: relevance of damage control orthopedic surgery. AB - Information illustrating the benefits of fracture stabilization after multiple trauma has been gathering for almost a century. At the turn of the last century, the introduction of the Thomas splint clearly demonstrated the importance of skeletal stabilization in the management of these patients. The introduction of standardized surgical treatment for fractures in the early 1950s is considered today as the turning point in the care of the polytraumatized patient. With the knowledge acquired, the application of early operative fixation of fractures in severely injured patients in the 1980s has yielded to the concept of early total care of all fractures. Yet, in distinct patient subgroups with severe thoracic injuries and very high injury severity scores, this concept has been associated with adverse outcomes. Therefore, in a further era that began in the 1990s, a different approach has been favored for these subgroups. It recommends early (initial) temporary stabilization followed by secondary definitive osteosynthesis of major fractures in patients at high risk of developing systemic complications. In the last decade, attempts have been made to determine which patients benefit from early total care and which ones should undergo a secondary definitive approach. This manuscript provides a historical overview on the changing treatment of fractures and summarizes the evolution of "damage control orthopedic surgery." PMID- 12095591 TI - Multimodal strategies to improve surgical outcome. AB - OBJECTIVE: To evaluate the effect of modifying perioperative care in noncardiac surgical patients on morbidity, mortality, and other outcome measures. BACKGROUND: New approaches in pain control, introduction of techniques that reduce the perioperative stress response, and the more frequent use of minimal invasive surgical access have been introduced over the past decade. The impact of these interventions, either alone or in combination, on perioperative outcome was evaluated. METHODS: We searched Medline for the period of 1980 to the present using the key terms fast track surgery, accelerated care programs, postoperative complications and preoperative patient preparation; and we examined and discussed the articles that were identified to include in this review. This information was supplemented with our own research on the mediators of the stress response in surgical patients, the use of epidural anesthesia in elective operations, and pilot studies of fast track surgical procedures using the multimodality approach. RESULTS: The introduction of newer approaches to perioperative care has reduced both morbidity and mortality in surgical patients. In the future, most elective operations will become day surgical procedures or require only 1 to 2 days of postoperative hospitalization. Reorganization of the perioperative team (anesthesiologists, surgeons, nurses, and physical therapists) will be essential to achieve successful fast track surgical programs. CONCLUSIONS: Understanding perioperative pathophysiology and implementation of care regimes to reduce the stress of an operation, will continue to accelerate rehabilitation associated with decreased hospitalization and increased satisfaction and safety after discharge. Developments and improvements of multimodal interventions within the context of "fast track" surgery programs represents the major challenge for the medical professionals working to achieve a "pain and risk free" perioperative course. PMID- 12095592 TI - The use of the Kugel mesh in ventral hernia repairs. AB - BACKGROUND: The management of ventral hernias is a common problem. Goals of hernia repair include reduction of the contents, reperitonealization of the abdomen, repair of the fascial defect, and restoration of the normal abdominal wall contour. Repairs frequently require the use of mesh to reduce the incidence of recurrence. A variety of repair techniques have been described, often associated with significant pain, prolonged recovery, and complications. METHODS: We describe a technique using a partial component separation, reperitonealization of the abdominal cavity with the hernia sack, and insertion of the Kugel mesh for bridging the gap. RESULTS: We have used this technique on 65 ventral hernias. Fifty-two patients went home from the recovery room and 11 stayed less than 23 hours. One patient had a wound infection and 2 had seromas. There have been no recurrences to date. CONCLUSIONS: Repair of ventral hernias with the Kugel mesh using a partial component separation is safe, effective and durable. PMID- 12095593 TI - Adenoid cystic breast cancer. AB - BACKGROUND: Adenoid cystic carcinoma is a rare type of breast cancer that is generally reported in individual case reports or as series from major referral centers. To characterize early diagnostic criteria for adenoid cystic carcinoma and to determine whether breast-preserving surgery with radiotherapy is as effective as mastectomy for eradicating the disease, we reviewed clinical records of a large series of patients treated for adenoid cystic carcinoma of the breast at a large health maintenance organization (HMO) that includes primary care facilities and referral centers. METHODS: Using the data bank of the Northern California Cancer Registry of the Kaiser Permanente Northern California Region (KPNCR), we retrospectively reviewed medical records of patients treated for adenoid cystic carcinoma of the breast. Follow-up also was done for these patients. RESULTS: Adenoid cystic carcinoma of the breast was diagnosed in 22 of 27,970 patients treated for breast cancer at KPNCR from 1960 through 2000. All 22 patients were female and were available for follow-up. Mean age of patients at diagnosis was 61 years (range, 37 to 94 years). In 17 (77%) of the women, a lump in the breast led to initial suspicion of a tumor; in 4 (23%) of the 22 patients, mammography led to suspicion of a tumor. Median tumor size was 20 mm. Pain was a prominent symptom. Surgical management evolved from radical and modified radical mastectomy to simple mastectomy or lumpectomy during the study period, during which time 1 patient died of previous ordinary ductal carcinoma of the contralateral breast, and 7 died of unrelated disease. At follow-up, 12 of the 13 remaining patients were free of disease; 1 patient died of the disease; and 1 patient remained alive despite late occurrence of lymph node and pulmonary metastases. CONCLUSIONS: Whether breast-preserving surgery with radiotherapy is as effective as mastectomy for treating adenoid cystic carcinoma of the breast has not been determined. PMID- 12095594 TI - Conservative surgery and radiation therapy for macroscopically multiple ipsilateral invasive breast cancers. AB - BACKGROUND: The presence of macroscopically multiple ipsilateral invasive breast cancer (MMIIBC) has been considered a contraindication for breast conservation. Early series reported high rates of local recurrence. A treatment regimen was developed to accommodate patient requests for breast conservation in MMIIBC. METHODS: We reviewed medical records of the 15 MMIIBC patients who underwent partial mastectomy followed by radiation between 1989 and 1997. All patients had 2 or more separate macroscopic tumors greater than 2 mm in diameter. After tumor excision, all specimens were evaluated; the protocol required surgical margins of at least 2 mm. RESULTS: As of June 2000 (median follow-up 76 months), 14 patients (93%) were alive without evidence of disease. One patient died of systemic disease without local recurrence. CONCLUSIONS: In selected cases, the combination of breast conservative surgery and radiation therapy with systemic therapy results in acceptable local-regional control. Patients who present with MMIIBC with clear surgical margins should be considered for breast conservation. PMID- 12095595 TI - The effect of improving communication competency on the certifying examination of the American Board of Surgery. AB - BACKGROUND: Since 1991 the authors have offered a course that identifies content deficits, but only provides instruction directed at improving verbal and nonverbal behaviors. We report the outcome of this 10-year effort as success on the certifying examination of the American Board of Surgery between 1991 and 2001. METHODS: Sixteen 5-day courses were scheduled over 10 years. Participants included those who had not taken the oral examination or had failed at least once and invited senior faculty (n = 26). Sites were chosen to replicate the actual examination setting. RESULTS: There were 122 participants, with follow-up data available on 88. Success in the certifying examination after completing the course is 96 percent. CONCLUSIONS: Evaluation of communication deficits and training to improve them is strongly associated with success. Clearly, this course is effective at identifying communication behaviors that are interfering with success on the certifying examination of the American Board of Surgery. PMID- 12095596 TI - The medical student as patient navigator as an approach to teaching empathy. AB - BACKGROUND: When empathy was presented in a lecture, medical students did not appreciate the topic or have opportunities to practice empathic techniques. The Patient Navigator Project was designed to foster understanding of a patient's experiences and encourage empathic communication. METHODS: The project uses experiential learning in an outpatient setting in the first year of training. Each student was to "shadow" a patient during a visit to a surgical oncologist and could observe the patient throughout treatment. Patient consent was obtained. Assignments were made before the appointments. Afterward, students met in small groups to reflect on their experiences. RESULTS: Patients were willing to let medical students observe their care. Most students reported a positive learning experience and that they learned to see patients as people, not as numbers or diseases. CONCLUSIONS: Requiring individualized patient-centered activities in the preclinical curriculum is challenging owing to the large number of students and the unpredictable nature of patients. PMID- 12095597 TI - Are multiple objective measures of student performance necessary? AB - BACKGROUND: This study examines the effect of using multiple modalities to evaluate medical students. METHODS: Thirty-four students were evaluated by a complex model utilizing National Board of Medical Examiners (NBME) shelf examination, Objective Structured Clinical Examination (OSCE), Computer Patient Simulation (CPS), faculty and peer evaluation. Results were compared with a traditional model based on NBME and faculty evaluation alone. RESULTS: Reliability (coefficient alpha) of the complex and traditional models were 0.72 and 0.47, respectively. Item correlations suggested that NBME was most discriminating (r = 0.75), followed by OSCE (r = 0.52), peer evaluation (r = 0.43), CPS (r = 0.39), and faculty evaluation (r = 0.32). Rank order correlation (Spearman's rho) between scores calculated using each model was 0.87. CONCLUSIONS: Although the complex model has improved reliability, both models rank students similarly. However, neither model fully captures and reflects the information provided by each of the specific evaluation methods. PMID- 12095598 TI - Gastrinoma. PMID- 12095599 TI - The role of laparoscopic cholecystectomy in the management of acute cholecystitis in patients with sickle cell disease. AB - BACKGROUND: As emergency surgery in sickle cell disease patients is associated with high morbidity, the aim of the study was to assess the safety of laparoscopic cholecystectomy in the acute state for these patients. METHODS: Over a 5-year period, April 1994 till December 1998, 35 sickle cell patients with acute cholecystitis had laparoscopic cholecystectomy within the first 5 days of presentation. A retrospective study of these was undertaken. RESULTS: Thirty-five patients were diagnosed as having acute cholecystitis with sickle cell disease. There were 26 female and 9 male patients; 5 patients needed preoperative and 1 patient needed postoperative endoscopic retrograde cholangiopancreatography. Twenty-seven patients needed simple transfusion and 8 needed partial exchange; conversion was necessary in two cases (5.7%). The mean hospital stay was 5.3 days and the complication rate was 17.5%. CONCLUSIONS: Because of the lack of significant complications, we believe that laparoscopic cholecystectomy for acute cholecystitis is safe and recommended in experienced hands with adequate preoperative preparation for patients with sickle cell disease. PMID- 12095600 TI - Intraoperative neuromonitoring of surgery for benign goiter. AB - BACKGROUND: Recurrent laryngeal nerve (RLN) palsy is one of the most serious complications in thyroid surgery. No prospective studies are available that evaluate if the additional use of intraoperative neuromonitoring reduces the rate of RLN palsy. METHODS: Between January 1 and December 31, 1998, surgery for histologically benign goiter with intraoperative identification with and without additional intraoperative RLN neuromonitoring was performed on 4,382 patients in 45 hospitals. Data were collected prospectively by questionnaire. RESULTS: The rate of transient and permanent RLN palsy based on nerves at risk were 1.4% and 0.4% with intraoperative neuromonitoring. These rates were significantly lower (P <0.05) compared with intraoperative visual RLN identification without intraoperative neuromonitoring which resulted in rates of 2.1% and 0.8%, respectively. A multivariate logistic regression analysis confirmed that the use of intraoperative neuromonitoring decreases the rate of postoperative transient (P <0.008) and permanent (P <0.004) RLN palsies as an independent factor by 0.58 and 0.30, respectively. CONCLUSIONS: Intraoperative neuromonitoring of the RLN in thyroid surgery is recommended because of significantly lower rates of transient and permanent RLN palsy rates in comparison with conventional RLN identification. PMID- 12095601 TI - Analysis of the relationships between clinicopathologic factors and survival time in intrahepatic cholangiocarcinoma. AB - BACKGROUND: This study elucidated the relationships between various clinicopathologic factors and the outcome of patients with intrahepatic cholangiocarcinoma (ICC) treated by hepatic resection. METHODS: A total of 37 ICC patients were treated by hepatic resection in our department between March 1979 and March 2001. Eleven clinicopathological variables (age, sex, preoperative jaundice, operative curability, number of tumors, UICC [Union Internationale Contre le Cancer] pT factor, UICC pN factor, UICC pM factor, histological tumor type, 10-year period during which they initially examined, and adjuvant therapy) were selected for univariate and multivariate analysis to evaluate their influence on the outcome. RESULTS: The actuarial 1-, 3-, and 5-year survival rates in the 37 resected cases were 54.1%, 34.0%, and 23.9%, respectively. The stage of the ICC influenced their overall survival rate. The univariate analysis revealed that curative resection (P = 0.0018), UICC pT factor (P = 0.0445), pN factor (P = 0.0029), pM factor (P = 0.0022), and histological type (P = 0.0030) were significant risk factors for survival. Multivariate analysis revealed that noncurative resection, lymph node metastasis, and less differentiated histological type were significant risk factors for poor outcome. All 6 of the 37 patients who survived more than 5 years had undergone curative resection, all of their tumors were well differentiated, and none had lymph node metastasis. CONCLUSIONS: Curative surgical resection remains the only effective approach to the treatment of ICC. Extensive resection is not indicated if lymph node metastasis can be identified preoperatively or intraoperatively. Current adjuvant therapy is ineffective, and it will be necessary to assess the efficacy of new adjuvant therapy strategies or the addition of new agents in terms of the outcome of ICC. PMID- 12095602 TI - Nuclear morphometric analysis of T2 lesions of the rectum--a simple, reproducible method for predicting malignancy potential. AB - BACKGROUND: Using computerized nuclear morphometry, we searched for common, objective parameters for use in predicting potential malignancy of primary T2 tumors of the rectum. METHODS: In 119 surgically resected T2 rectal cancers, the following parameters of the cancer cell nuclei were microscopically measured: mean nuclear area (NA), mean perimeter (PM), largest-to-smallest diameter ratios (LS), circularity factor (CF), and coefficient of variation of nuclear area (NACV). The calculated parameters were retrospectively analyzed and compared with patients' outcome. RESULTS: NA, PM, and NACV correlated with recurrence. NA and NACV were significantly associated with disease-free survival. High NACV correlated with lymph node metastasis. In node-negative cases, high NACV correlated with recurrence and tended to be associated with poor prognosis. CONCLUSIONS: These findings suggest that computerized morphometry is useful for predicting malignancy potential of T2 lesions of the rectum. PMID- 12095603 TI - Subtotal gastrectomy for cancer located in the greater curvature of the middle stomach with prevention of the left gastric artery. AB - BACKGROUND: A novel distal subtotal gastrectomy was performed in 5 patients with macroscopically node-positive gastric cancer located in the greater curvature of the middle stomach. In these patients, total gastrectomy or standard distal subtotal gastrectomy has been typically performed. In these typical gastrectomies, the hepatic and the coeliac branches of the vagi are removed en bloc with the left gastric artery and the whole of the lesser omentum because the lymphatics along the left gastric artery are in the lower stream-regions of lymph flow from the cancer and metastases exist potentially. METHODS: During novel distal subtotal gastrectomy the activated carbon method confirmed that the lymphatics along the ascending branch of the left gastric artery were not in the lower stream-region of lymph flow from the cancer. Then, we preserved the hepatic and coeliac branches of the vagi as well as the ascending branch of the left gastric artery and the upper part of the lesser omentum. The other arteries feeding the stomach were removed with the surrounding lymphatics. In novel distal subtotal gastrectomy the remnant stomach was fed only by the ascending branch of the left gastric artery, while in standard distal subtotal gastrectomy the remnant stomach was fed by the short gastric arteries. CONCLUSIONS: Although further examinations are necessary, novel distal subtotal gastrectomy may have superior merit such as good function of gallbladder because of the preservation of the vagal nerve system, compared with total gastrectomy or standard distal subtotal gastrectomy. PMID- 12095604 TI - Colonic anastomosis with the nickel-titanium temperature-dependent memory-shape device. AB - BACKGROUND: This study was prompted by our successful experience with a compression anastomosis clip (CAC) in an animal model. METHODS: The study sample included 20 patients scheduled for colonic resections, of whom 10 underwent anastomosis with the CAC and 10 with staplers. RESULTS: Neither group had anastomotic or other complications, except for 1 patient in the CAC group in whom a subphrenic infected hematoma developed after left hemicolectomy with splenectomy. CONCLUSIONS: Our study demonstrates that this CAC is safe and simple to use, coming close to the "no touch surgery concept" and is of low cost compared with the staples used today. PMID- 12095605 TI - The effect of robotic assistance on learning curves for basic laparoscopic skills. AB - BACKGROUND: We hypothesized that laparoscopic tasks performed with ZEUS robotic assistance would be done with greater precision and with a different learning curve than when performed in a standard laparoscopic trainer. METHODS: Participants were divided into the surgically experienced (n = 11) and the surgically naive (n = 17). Two laparoscopic tasks (bead transfer and rope pass) were repeated for five repetitions. RESULTS: For all drills and participants, completion time and error rate decreased across the five repetitions for each platform. Precision averaged 97% for both platforms over all drills. For both groups, completion time for tasks was shorter on the laparoscopic platform. ZEUS allowed for greater consistency in performance. CONCLUSIONS: Compared with performance on a standard laparoscopic trainer, robotic assistance allows for increasing speed and consistency while maintaining precision over multiple repetitions. Understanding how robotics affects learning curves will allow for modifications in the training experience with this new technology. PMID- 12095609 TI - Ethanol cycle in an ethanologenic bacterium. AB - A novel redox cycle is suggested, performing interconversion between acetaldehyde and ethanol in aerobically growing ethanologenic bacterium Zymomonas mobilis. It is formed by the two alcohol dehydrogenase (ADH) isoenzymes simultaneously catalyzing opposite reactions. ADH I is catalyzing acetaldehyde reduction. The local reactant ratio at its active site probably is shifted towards ethanol synthesis due to direct channeling of NADH from glycolysis. ADH II is oxidizing ethanol. The net result of the cycle operation is NADH shuttling from glycolysis to the membrane respiratory chain, and ensuring flexible distribution of reducing equivalents between the ADH reaction and respiration. PMID- 12095608 TI - Does chloroquine really act through oxidative stress? AB - To assess whether molecular oxygen and oxidative stress contribute to chloroquine activity, we cultivated strains of Plasmodium falciparum in erythrocytes with carboxyhemoglobin and an atmosphere containing 2% CO, 5% CO(2) and 93% N(2). Results indicate that, contrary to common belief, oxygen is not involved in the activity of chloroquine. Reactive radicals formation is suggested. PMID- 12095610 TI - Amino acid determinants of alpha-synuclein aggregation: putting together pieces of the puzzle. AB - Parkinson's disease is the second most common neurodegenerative disease, and results from loss of dopaminergic neurons in the substantia nigra. The aggregation and fibrillation of alpha-synuclein in the form of intracellular proteinaceous aggregates (Lewy bodies and Lewy neurites) have been implicated as a causative factor in this disease, as well as in several other neurodegenerative disorders, including dementia with Lewy bodies, Lewy body variant of Alzheimer's disease, multiple system atrophy and Hallervorden-Spatz disease. Thus, the aggregated forms of alpha-synuclein play a crucial role in the pathogenesis of the synucleinopathies. However, the molecular mechanisms underlying alpha synuclein aggregation into specific filamentous inclusions remained unknown until recently. Data on the aggregation and fibrillation properties of human alpha-, beta- and gamma-synucleins, mouse alpha-synuclein and familial Parkinson's disease mutants of human alpha-synuclein (A30P and A53T) are analyzed in order to shed light on the amino acid determinants of synuclein aggregation. PMID- 12095611 TI - Plasticity of proton pathways in haem-copper oxygen reductases. AB - Oxygen reductases are the final enzymes in the aerobic respiratory chains catalysing the reduction of dioxygen to water, with the concomitant translocation of protons through the bacterial cytoplasmatic or mitochondrial membranes. Most of these enzymes belong to the family of haem-copper oxygen reductases. Intraprotein proton-conducting pathways are needed for the chemical reaction and for the translocated protons. Based on sequence and structural analyses, and site directed mutagenesis, two proton channels were established for the mitochondrial like oxygen reductases. However, the amino acid residues forming these channels are not conserved among the family members. Most importantly, many oxygen reductases do not contain ionisable amino acid residues in the putative proton pathways nor in alternative positions. The diversity of channels in haem-copper oxygen reductases exemplifies the plasticity of proton pathways that occurred throughout evolution, and strongly suggests a substantial role for water as the main proton carrier. PMID- 12095613 TI - Discriminant analysis to evaluate clustering of gene expression data. AB - In this work we present a procedure that combines classical statistical methods to assess the confidence of gene clusters identified by hierarchical clustering of expression data. This approach was applied to a publicly released Drosophila metamorphosis data set [White et al., Science 286 (1999) 2179-2184]. We have been able to produce reliable classifications of gene groups and genes within the groups by applying unsupervised (cluster analysis), dimension reduction (principal component analysis) and supervised methods (linear discriminant analysis) in a sequential form. This procedure provides a means to select relevant information from microarray data, reducing the number of genes and clusters that require further biological analysis. PMID- 12095612 TI - The expression of native and oxidized LDL receptors in brain microvessels is specifically enhanced by astrocytes-derived soluble factor(s). AB - Ex vivo rat brain microvessels express receptors for native as well as for oxidized low-density lipoproteins. In brain microvessels-derived endothelial cells, the expression levels of both receptors were enhanced by co-cultivation with rat astrocytes, even in the absence of actual contact between the two cell types, suggesting a soluble factor(s)-based mechanism of induction. No modulation effect could be evidenced in a heterologous cellular system. Since both receptors were found to be expressed also in astrocytes, these cells are likely to contribute substantially to the lipoprotein management at the blood-brain barrier and in the brain compartment. PMID- 12095614 TI - Direct addition of BimL to mitochondria does not lead to cytochrome c release. AB - Pro-apoptotic members of the Bcl-2 family can be subdivided in two classes according to their structure: a group including Bax, Bak, and Bok that display Bcl-2 homology (BH) 1, BH2 and BH3 domains and a second group including Bid (BH3 interacting domain death agonist), Bad, Bim (Bcl-2 interacting mediator of cell death) and several others that contain only a BH3 domain, the BH3-only proteins. The BH3-only proteins have been proposed to activate pro-apoptotic members of the Bax subfamily to trigger a mitochondrial pathway that leads to the release of cytochrome c and other apoptogenic factors. Here we report that the mechanism of action of Bim is different from that of Bid. Although overexpression of Bid or Bim in cells leads to cytochrome c release, only Bid is able to trigger the release of cytochrome c through Bax activation when added directly to isolated mitochondria. Bim(L), although unable to activate Bax, can directly inhibit Bcl-2 or Bcl-x(L). Our data suggest two functional classes of BH3-only proteins: those such as Bid which directly activate Bax-like proteins leading to mitochondrial membrane permeability and apoptosis and those such as Bim which inhibit anti apoptotic proteins and render the cells more susceptible to apoptogenic stimuli. PMID- 12095615 TI - Characterization of bacterial homocitrate synthase involved in lysine biosynthesis. AB - In Thermus thermophilus homocitrate synthase (HCS) catalyzes the initial reaction of lysine biosynthesis through alpha-aminoadipic acid, synthesis of homocitrate from 2-oxoglutarate and acetyl-CoA. HCS is strongly inhibited by lysine, indicating that the biosynthesis is regulated by the endproduct at the initial reaction in the pathway. HCS also catalyzes the reaction using oxaloacetate in place of 2-oxoglutarate as a substrate, similar to citrate synthase in the tricarboxylic acid cycle. Several other properties of Thermus HCS and an evolutionary relationship of the biosynthetic pathway in the bacterium to other metabolic pathways are also described. PMID- 12095617 TI - The gene expression and deficiency phenotypes of Cockayne syndrome B protein in Caenorhabditis elegans. AB - The Caenorhabditis elegans Cockayne syndrome B protein homologue is encoded by 10 exons of the predicted open reading frame F53H4.1. The gene is expressed in germ cells and all somatic cells of the embryonic to adult stage. Although the gene expression was ubiquitous, its expression level was relatively higher in dividing cells and cells that play fundamental roles in essential physiological functions such as feeding, sensation, and reproduction. RNA interference of the gene hypersensitized C. elegans to UV radiation, as observed in enhanced germ cell proliferation arrest and apoptosis, and increased embryonic lethality, suggesting its role in nucleotide excision repair. PMID- 12095616 TI - Naturally occurring heterologous trans-splicing of adenovirus RNA with host cellular transcripts during infection. AB - The impact of viral infection on normal host RNA processing remains largely unexplored. We postulated that the high abundance of virally derived nuclear RNA in infected cells could impact host cell RNA splicing and viability. To test for aberrant RNA splicing we examined trans-splicing following infection with the replication-competent adenovirus mutant d11520 that lacks E1B 55 kDa protein. Trans-splicing was observed between viral RNA and several cellular precursor mRNAs, including beta-actin and glyceraldehyde-3-phosphate dehydrogenase. Using a tetracycline-inducible model system simulating viral trans-splicing activity we observed that overexpression of a trans-splicing RNA specifically inhibited cell proliferation. These results demonstrate that heterologous trans-splicing occurs naturally during adenovirus infection and suggest that trans-splicing may contribute to disruption of cell function. PMID- 12095618 TI - Transposon-like Correia elements: structure, distribution and genetic exchange between pathogenic Neisseria sp. AB - Correia elements are a prominent feature of all four Neisseria genome sequences. We report an in silico analysis of the structure and genomic distribution of these elements and some preliminary biochemical data. Correia elements fall into four major families, distinguished by a 50 bp internal deletion and five point mutations. The elements resemble a transposon with 25 bp inverted repeats and a TA duplication at the target site. Within the element there is a functional integration host factor binding site. The genomic distribution of Correia elements is essentially random except for some small Correia-less regions apparently acquired by horizontal transfer. Phylogenetic analysis suggests that their presence predates the divergence of Neisseria meningitidis and Neisseria gonorrhoeae. PMID- 12095619 TI - Suppression of DTT-induced aggregation of abrin by alphaA- and alphaB crystallins: a model aggregation assay for alpha-crystallin chaperone activity in vitro. AB - The eye lens small heat shock proteins (sHSP), alphaA- and alphaB-crystallins, have been shown to function like molecular chaperones, both in vitro and in vivo. It is essential to assess the protective effect of alphaA- and alphaB-crystallins under native conditions to extrapolate the results to in vivo conditions. Insulin and alpha-lactalbumin have widely been used to investigate the chaperone mechanism of alpha-crystallin under native conditions. Due to its smaller size, insulin B-chain may not represent the binding of putative physiological substrate proteins. As it stands, the aggregation of alpha-lactalbumin and binding of alpha crystallin to it varies under different experimental conditions. Abrin, a ribosome inactivating protein isolated from the seeds of Abrus precatorius, consists of a 30 kDa A-chain and a lectin-like B-chain of 33 kDa joined by a single disulfide bond. Reduction of the disulfide link between the two chains of abrin leads to the aggregation of the B-chain. In this study, we demonstrate that dithiothreitol (DTT)-induced aggregation of abrin B-chain could be monitored by light scattering similar to that of insulin. Moreso, this process could be suppressed by recombinant human alphaA- and alphaB-crystallins in a concentration dependent manner, notably by binding to aggregation prone abrin B-chain. SDS-PAGE and HPLC gel filtration analysis indicate that there is a soluble complex formation between alpha-crystallin and abrin B-chain. Interestingly, in contrast to insulin, there is no significant difference between alphaA- and alphaB crystallin in suppressing the aggregation of abrin B-chain at two different temperatures (25 and 37 degrees C). HSP26, an another small heat shock/alpha crystallin family protein, was also able to prevent the DTT-induced aggregation of abrin. These results suggest that due to relatively larger size of its B-chain (33 kDa), compared to insulin B-chain (about 3 kDa), abrin may serve as a better model substrate for in vitro chaperone studies of alpha-crystallin and as well as other sHSP. PMID- 12095620 TI - PACAP protects neuronal PC12 cells from the cytotoxicity of human prion protein fragment 106-126. AB - Misfolding of the prion protein yields amyloidogenic isoforms, and it shows exacerbating neuronal damage in neurodegenerative disorders including prion diseases. Pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) potently stimulate neuritogenesis and survival of neuronal cells in the central nervous system. Here, we tested these neuropeptides on neurotoxicity in PC12 cells induced by the prion protein fragment 106-126 [PrP (106-126)]. Concomitant application of neuropeptide with PrP(106-126) (5x10(-5) M) inhibited the delayed death of neuron-like PC12 cells. In particular, PACAP27 inhibited the neurotoxicity of PrP(106-126) at low concentrations (>10(-15) M), characterized by the deactivation of PrP(106-126) stimulated caspase-3. The neuroprotective effect of PACAP27 was antagonized by the selective PKA inhibitor, H89, or the MAP kinase inhibitor, U0126. These results suggest that PACAP27 attenuates PrP(106-126)-induced delayed neurotoxicity in PC12 cells by activating both PKA and MAP kinases mediated by PAC1 receptor. PMID- 12095621 TI - Quantification of NF1 transcripts reveals novel highly expressed splice variants. AB - Previously, we have shown that the NF1 gene gives rise to multiple novel splice variants. In the present study, nine NF1 variants were quantified by real-time PCR in various human tissues. Some of these variants were expressed at low to moderate low levels and possible implications of these findings are discussed. Interestingly, two variants (NF1-DeltaE4b and NF1-DeltaE43) were shown to be highly expressed in specific tissues. NF1-DeltaE43 lacks a nuclear targeting sequence and might be functionally different from full-length NF1. These novel NF1 splice variants might expand our understanding of the role of neurofibromin. PMID- 12095622 TI - Temperature-induced selective death of the C-domain within angiotensin-converting enzyme molecule. AB - Somatic angiotensin-converting enzyme (ACE) consists of two homologous domains, each domain bearing a catalytic site. Differential scanning calorimetry of the enzyme revealed two distinct thermal transitions with melting points at 55.3 and 70.5 degrees C. which corresponded to denaturation of C- and N-domains, respectively. Different heat stability of the domains underlies the methods of acquiring either single active N-domain or active N-domain with inactive C-domain within parent somatic ACE. Selective denaturation of C-domain supports the hypothesis of independent folding of the two domains within the ACE molecule. Modeling of ACE secondary structure revealed the difference in predicted structures of the two domains, which, in turn, allowed suggestion of the region 29-133 in amino acid sequence of the N-part of the molecule as responsible for thermostability of the N-domain. PMID- 12095623 TI - Modification of heme c binding motifs in the small subunit (NrfH) of the Wolinella succinogenes cytochrome c nitrite reductase complex. AB - The two multiheme c-type cytochromes NrfH and NrfA form a membrane-bound complex that catalyzes menaquinol oxidation by nitrite during respiratory nitrite ammonification of Wolinella succinogenes. Each cysteine residue of the four NrfH heme c binding motifs was individually replaced by serine. Of the resulting eight W. succinogenes mutants, only one is able to grow by nitrite respiration although its electron transport activity from formate to nitrite is decreased. NrfH from this mutant was shown by matrix-assisted laser desorption/ionization mass spectrometry to carry four covalently bound heme groups like wild-type NrfH indicating that the cytochrome c biogenesis system II organism W. succinogenes is able to attach heme to an SXXCH motif. PMID- 12095624 TI - Transcriptional regulation of nuclear orphan receptor, NOR-1, by Ca(2+)/calmodulin-dependent protein kinase cascade. AB - We found that transcription of NOR-1 (neuron-derived orphan receptor-1) is induced by the Ca(2+)/calmodulin-dependent protein kinase IV (CaM-KIV) cascade by differential hybridization screening of a cDNA array using probes prepared from SH-SY5Y cells infected with recombinant adenoviruses expressing Ca(2+)/CaM independent mutants of CaM-KIV and CaM-K kinase (CaM-KK). Induction of NOR-1 mRNA expression by the CaM-KIV cascade was confirmed by RT-PCR. Promoter analyses indicate that CaM-KIV cascade response element is located between -162 bp and -42 bp in the 1.7 kb NOR-1 promoter containing triple cAMP response elements. Disruption of each element significantly reduced the promoter activity, indicating the direct transcriptional regulation of NOR-1 by CaM-KIV cascade. PMID- 12095625 TI - Inhibition of ubiquitin-dependent proteolysis by a synthetic glycine-alanine repeat peptide that mimics an inhibitory viral sequence. AB - The glycine-alanine repeat (GAr) of the Epstein-Barr virus nuclear antigen-1 is a cis-acting transferable element that inhibits ubiquitin/proteasome-dependent proteolysis in vitro and in vivo. We have here examined the effect of a synthetic 20-mer GAr oligopeptide on the degradation of iodinated or biotin labeled lysozyme in a rabbit reticulocyte lysates in vitro assay. Micromolar concentrations of the GA-20 peptide inhibited the hydrolysis of lysozyme without significant effect on ubiquitination. Addition of the peptide did not inhibit the hydrolysis of fluorogenic substrate by purified proteasomes and did not affect the ubiquitination of lysozyme. An excess of the peptide failed to compete for binding of a synthetic tetra-ubiquitin complex to the S5a ubiquitin-binding subunit of the 19S regulator, confirming that the GAr does not block the access of ubiquitinated substrates to the proteasome. Our data suggest that the GAr may act by destabilizing the interaction of ubiquitinated substrates with the proteasome and promote the premature release of the substrate. PMID- 12095626 TI - Increased early atherogenesis in young versus old hypercholesterolemic rabbits by a mechanism independent of arterial cell proliferation. AB - We sought to determine the relative importance of aging and hypercholesterolemia on atherosclerosis. Although plasma cholesterol levels increased similarly in young and old rabbits fed an atherogenic diet for 2 months, aortic atherosclerotic lesions were more prominent in young animals. This finding was associated with an age-dependent reduction in the DNA-binding activity of the proinflammatory nuclear factor kappaB (NF-kappaB) in aortic tissue. Atherosclerotic lesions consisted mostly of macrophages, which displayed a similar proliferative response in both age groups. Independently of the age, medial cell proliferation was low and increased as a function of intimal lesion size. Thus, higher atherogenicity in young rabbits exposed to extreme hypercholesterolemia compared to old counterparts is associated with higher activity of NF-kappaB in the juvenile vessel wall without apparent age-dependent changes in arterial cell proliferation. PMID- 12095627 TI - Protein disulfide isomerase of Toxoplasma gondii is targeted by mucosal IgA antibodies in humans. AB - Mass spectrometric analysis identified a 49 kDa antigen from Toxoplasma gondii as protein disulfide isomerase (PDI). This antigen is generally recognized by IgA in tears of healthy humans. We determined the complete open reading frame and expressed PDI recombinantly. Recombinant PDI was recognized by IgA in human tears known to contain antibodies specific for the 49 kDa antigen. High expression level and similarity to other protozoan PDIs suggest that T. gondii PDI might be a suitable target for recently described anti-protozoan drugs. PDI-specific antibodies clearly constitute part of the mucosal antibody repertoire possibly involved in defence against parasites. PMID- 12095628 TI - A polygalacturonase of animal origin isolated from the root-knot nematode Meloidogyne incognita. AB - The first animal polygalacturonase (PG, EC 2.1.15) encoding cDNA, Mi-pg-1, was cloned from the plant parasitic nematode Meloidogyne incognita. The enzymatic activity of MI-PG-1 was confirmed after heterologous expression in Escherichia coli. The presence of a predicted signal peptide on the MI-PG-1 sequence together with the specific localization of the transcripts of the Mi-pg-1 gene in the oesophageal glands of infective juveniles imply that MI-PG-1 could be secreted into plant tissues. The potential role of MI-PG-1 in parasitism is discussed. PMID- 12095629 TI - Control of capillary formation by membrane-anchored extracellular inhibitor of phospholipase A(2). AB - Secretory phospholipase A(2) (sPLA(2)) has been reported to be involved in cell proliferation in general and in endothelial cell migration, processes required for capillary formation. Subsequently, we examined the potential control of angiogenesis by sPLA(2) inhibition, using a cell-impermeable sPLA(2) inhibitor composed of N-derivatized phosphatidyl-ethanolamine linked to hyaluronic acid. This inhibitor effectively inhibits the proliferation and migration of human bone marrow endothelial cells in a dose-dependent manner, and suppresses capillary formation induced by growth factors involved in vascularization of tumors and of atherosclerotic plaques. It is proposed that sPLA(2) inhibition introduces a novel approach in the control of cancer development and atherosclerosis. PMID- 12095630 TI - Evidence for two interaction regions for phosphatidylinositol(4,5)-bisphosphate on mammalian profilin I. AB - The binding of phosphatidylinositol(4,5)-bisphosphate (PI(4,5)P(2)) to profilin at a region distinct from the actin interaction surface is demonstrated by experiments with covalently cross-linked profilin:beta-actin. The result is in agreement with observations made with several mutant profilins and provides strong evidence for two regions on mammalian profilin mediating electrostatic interaction with phosphatidylinositol lipids; one close to the binding site for poly(L-proline), and one partially overlapping with the actin-binding surface. Congruent with this, two plant profilins, which have a reduced number of positive amino acids in one of these regions, displayed a dramatically lower binding to PI(4,5)P(2) compared to human profilin I. PMID- 12095632 TI - Role of intracellular loops of cannabinoid CB(1) receptor in functional interaction with G(alpha16). AB - The cannabinoid CB(1) but not the CB(2) receptor was demonstrated to couple via G(alpha16) to activate phospholipase C after co-expression in COS7 cells. Chimeric CB(1)/CB(2) receptors were used as a model to study receptor-G(alpha16) interaction. Sequences of the second and third intracellular loops and the carboxy-terminus were substituted from the CB(1) into the CB(2) receptor. Only the triple mutant with all three regions replaced activated phospholipase C to a similar extent as the CB(1) receptor, suggesting that all three intracellular regions are required for interacting with G(alpha16). Several sub-domains within the third intracellular loop were identified for receptor-G(alpha16) interaction. PMID- 12095631 TI - Hypoxia activates a background conductance in freshly isolated pulmonary arterial endothelial cells of the rat. AB - Utilising the patch-clamp recording technique we have demonstrated for the first time the effects of hypoxia on the background current in pulmonary arterial endothelial cells. Electrophysiological studies revealed the presence of a novel oxygen-sensitive, non-selective cation conductance (I(NSC)) in these cells. The inward component of I(NSC) was significantly potentiated by hypoxia. Both the inward and outward components of I(NSC) were inhibited by both La(3+) and Gd(3+). Hypoxic activation of I(NSC) may provide an important Ca(2+) influx pathway essential for the release of a pulmonary-selective vasoconstrictor pivotal to the sustained phase of hypoxic pulmonary vasoconstriction. PMID- 12095634 TI - Flavinylation of the precursor of mitochondrial dimethylglycine dehydrogenase by intact and solubilised mitochondria. AB - The flavinylation and the presequence processing of the mitochondrial matrix enzyme dimethylglycine dehydrogenase (Me(2)GlyDH) were investigated with the reticulocyte lysate translated precursor (pMe(2)GlyDH) added to solubilised mitoplasts of rat liver mitochondria. The flavinylation of pMe(2)GlyDH was strictly dependent on the addition of mitochondrial protein(s), among which the mitochondrial flavinylation stimulating factor [Brizio C., et al. (2000) Eur. J. Biochem 267, 4346-4354], that actively promotes holo-Me(2)GlyDH formation. The precursor processing, that accompanies the biogenesis of the enzyme, was not required to allow the flavinylation to proceed. The comparison of the time course of the flavinylation and the processing of pMe(2)GlyDH demonstrated that the covalent attachment of the flavin moiety preceded the presequence processing by mitochondrial processing peptidase. PMID- 12095633 TI - Effects of salt and nickel ion on the conformational stability of Bacillus pasteurii UreE. AB - UreE, a urease accessory protein, is proposed to be a metallochaperone assisting the nickel incorporation into the urease active site. We investigated the effects of salt and nickel on the conformational stability of the UreE from Bacillus pasteurii (BpUreE), by circular dichroism (CD) and nuclear magnetic resonance spectroscopy accompanying a thermodynamic inspection. Far-UV CD spectra of BpUreE showed that both salt and nickel stabilized the ordered structure of the protein. The thermal denaturing of BpUreE showed a bimodal feature with an aggregation process before thermal unfolding. This thermally induced aggregation could be suppressed by the addition of salt up to 50 mM, and the further addition of salt increased the thermal resistance of the protein. The nickel addition also elevated the thermal resistance of BpUreE, although it could not prevent the aggregating process. Additionally, the stoichiometry of a specific nickel binding to BpUreE was revealed as one nickel per dimer. Altogether, the present results establish a rather detailed characterization of the thermostability and nickel binding property of BpUreE. PMID- 12095635 TI - Contribution of basic residues of the A helix of heparin cofactor II to heparin- or dermatan sulfate-mediated thrombin inhibition. AB - Inhibition of thrombin by heparin cofactor II (HCII) is accelerated 1000-fold by heparin or dermatan sulfate. To investigate the contribution of basic residues of the A helix of HCII to this activation, we constructed amino acid substitutions (K101Q, R103L, and R106L) by site-directed mutagenesis. K101Q greatly reduced heparin cofactor activity and required a more than 10-fold higher concentration of dermatan sulfate to accelerate thrombin inhibition compared with wild-type recombinant HCII. Thrombin inhibition by R106L was not significantly stimulated by dermatan sulfate. These results provide evidence that basic residues of the A helix of HCII (Lys(101) and Arg(106)) are necessary for heparin- or dermatan sulfate-accelerated thrombin inhibition. PMID- 12095636 TI - The action of N-acetylglucosaminyltransferase-V is prevented by the bisecting GlcNAc residue at the catalytic step. AB - Using a purified protein and bisected acceptor oligosaccharides, we demonstrate that N-acetylglucosaminyltransferase (GnT)-V transfers a N-acetylglucosamine residue via a beta1,6-linkage to the bisected oligosaccharides. We also kinetically characterized the substrate specificity of GnT-V with respect to the bisected oligosaccharide. Although the K(m) values for the bisected acceptors were comparable to that for a non-bisected acceptor, the V(max) values for the bisected acceptors were much lower than that for the non-bisected acceptor. These findings suggest that the acceptor specificity of GnT-V is determined by the catalytic process rather than by its binding to the substrate. It was also found that the presence of the 2-N-acetyl group in the bisecting monosaccharide moiety plays a critical role in determining the catalytic efficiency of the enzyme. PMID- 12095637 TI - Different roles of the IGF-I Ec peptide (MGF) and mature IGF-I in myoblast proliferation and differentiation. AB - The physiological function of a recently cloned splice variant of insulin-like growth factor-I (IGF-I; mechano growth factor (MGF)) was studied using an in vitro cell model. Unlike mature IGF-I, the distinct E domain of MGF inhibits terminal differentiation whilst increasing myoblast proliferation. Blocking the IGF-I receptor with a specific antibody indicated that the function of MGF E domain is mediated via a different receptor. The results provide a basis for localized tissue adaptation and helps explain why loss of muscle mass occurs in the elderly and in dystrophic muscle in which MGF production is markedly affected. PMID- 12095638 TI - Induction of p73beta by a naphthoquinone analog is mediated by E2F-1 and triggers apoptosis in HeLa cells. AB - Recently, p73 was identified as a structural and functional homolog of p53. The p73 protein activates the transcription of genes downstream of p53 and induces apoptosis when overexpressed in several cell lines, similar to the tumor suppressor p53. However, the extracellular stimuli and molecular mechanisms regulating p73 activity remain to be elucidated. In this paper, we present evidence that the naphthoquinone analog, 2,3-dichloro-5,8-dihydroxy-1,4 naphthoquinone (NA), is a novel apoptotic stimulus that induces p73beta expression. Treatment with NA induced the expression of p73beta mRNA and protein and its downstream genes, p21 and bax, in HeLa cells. Similar results were obtained in MCF7 cells (p53(+/+), p73(+/+)). In the MCF7 cells, p53 protein level was rather decreased by NA treatment. Overexpression of p73beta led to the apoptosis of HeLa cells and enhancement of NA-induced cell death. Expression of p73beta was mediated by E2F-1, which was activated via release from pRB after exposure of cells to NA. We additionally observed that overexpression of pRB inhibited NA-induced apoptosis. These results imply that p53-independent p73beta dependent p21 expression is involved in NA-induced apoptosis of HeLa cells. PMID- 12095639 TI - PUVA-induced apoptosis involves mitochondrial dysfunction caused by the opening of the permeability transition pore. AB - The mechanism of cell death was investigated in Jurkat cells exposed to the combination of psoralen and UVA irradiation (PUVA). Apoptosis was by far prevailing over necrosis and involved mitochondrial dysfunction. The collapse of mitochondrial membrane potential, appears to be caused by the opening of the mitochondrial permeability transition pore since its inhibitor, cyclosporin A, prevented mitochondrial dysfunction and largely attenuated apoptosis. Apoptosis also occurred in cells treated with the photoproducts generated by irradiating psoralen in vitro with an oxygen-dependent process. Thus, the involvement of reactive oxygen species in the onset of PUVA-induced apoptosis appears mostly related to psoralen photooxidation. PMID- 12095640 TI - Identification of the E2-binding residues in the N-terminal domain of E1 of a prokaryotic pyruvate dehydrogenase complex. AB - Pyruvate dehydrogenase (E1p) is one of the components of the pyruvate dehydrogenase multienzyme complex (PDHC). Previously, it was shown that the N terminal domain of E1p is involved in its binding to the core component (E2p) of PDHC. We constructed point mutations in this domain (D17Q, D17R, E20Q, E20R, D24Q and D24R) to identify the specific residues involved in these interactions. Kinetic and binding studies show that D17 is essential for the binding of E1p to E2p. D24 is involved in the binding, but not essential, whereas E20 is not involved. None of the mutations affects the folding or dimerisation of E1p. PMID- 12095641 TI - Association of HSP70 with endonucleases allows the expression of otherwise silent mutations. AB - A subpopulation of the 70 kDa heat shock protein (HSP70) found within the mitochondria of Saccharomyces cerevisiae functions as a stable binding partner of the endonuclease SceI. We have previously found that the SceI endonuclease monomer recognizes and cleaves a unique, 26 bp sequence in vitro. Dimerization with HSP70 changes the specificity of SceI, allowing it to cleave at multiple sequences. This study shows that SuvI, an ortholog of SceI isolated from a different yeast strain, contains two amino acid substitutions, yet it shows the same uni-site specificity in its monomeric form. Binding of HSP70 to the SuvI monomer confers multi-site specificity that is different from that exhibited by the HSP70/SceI heterodimer. Mutation of single residues of SceI to the corresponding residue in SuvI provides enzymes with specificities intermediate between SceI and SuvI when complexed with HSP70. These results suggest that HSP70 interaction with certain endonucleases allows the expression of otherwise silent mutations in them, causing a change in enzyme cleavage specificity. PMID- 12095642 TI - Gene expression profiling of cell lines derived from T-cell malignancies. AB - The expression profiles of eight cell lines derived from T-cell malignancies were compared to CD4-positive T-cells using cDNA microarray technology. Unsupervised hierarchical clustering of 4364 genes demonstrated substantial heterogeneity resulting in four distinct groups. While no genes were found to be uniformly up- or down-regulated across all cell lines, we observed 111 over-expressed genes (greater than two-fold) and 1118 down-regulated genes (greater than two-fold) in the lymphomas as a group when compared to CD4-positive T-cells. These included genes involved in cytokine signaling, cell adhesion, cytoskeletal elements, nuclear transcription factors, and known oncogenes and tumor suppressor genes. Quantitative fluorescent reverse transcription-polymerase chain reaction analysis demonstrated 70% concordance with the microarray results. While freshly isolated malignant cells may differ in their individual expression patterns relative to established cell lines from the same diagnoses, we feel that the variety of different lymphocytic cell lines that we examined provides a representative picture of the molecular pathogenesis of T-cell malignancies. PMID- 12095643 TI - Melatonin prevents the formation of pyrrolized proteins in human plasma induced by hydrogen peroxide. AB - This report presents a model of oxidative stress, which includes formation of pyrrolized proteins in human plasma. Pyrroles were determined using Ehrlich's reagent under acid conditions. Adduct formation in plasma proteins was induced by hydrogen peroxide (H(2)O(2)) in a dose-dependent manner. The simultaneous addition of melatonin to the incubation medium with 100 mM H(2)O(2) prevented the formation of Ehrlich adducts. This inhibitory effect of melatonin on adduct formation was significant at concentrations higher than 10 nM melatonin. When melatonin was added to the medium 60 min before adding H(2)O(2), no significant difference was observed. The effect of melatonin was also compared with other known antioxidants such as vitamin E (Trolox) and vitamin C; the order of potency of the antioxidants was melatonin>vitamin E>vitamin C. In conclusion, the results presented in this paper add new perspectives to other well known antioxidant properties of melatonin such as prevention of protein pyrrolization. PMID- 12095644 TI - Co-localization of 5-HT 2A -receptor- and GABA-immunoreactivity in neurones in the periaqueductal grey matter of the rat. AB - The dorsal half of the midbrain periaqueductal grey matter (dPAG) functions as a midbrain aversive system. Serotonin exerts anti-aversive effects in the dPAG that are mediated by actions at 5-hydroxytryptamine (5-HT)(1A)- and 5-HT(2A) receptors. Since at the cellular level, 5-HT(2A)-receptor activation has been shown to evoke excitatory responses in the dPAG, it is possible that anti aversive 5-HT(2A)-mediated effects are mediated indirectly by activation of inhibitory gamma-aminobutyric acid (GABA)ergic interneurones. In rats, immunoreactivity for 5-HT(2A)-receptors was present on the soma and dendrites of neurones throughout the PAG. Co-localization studies revealed that the majority (>90%) of 5-HT(2A)-receptor-labelled cells also showed immunoreactivity for GABA. These findings may provide an anatomical substrate for 5-HT(2A)-mediated anti aversive effects in the dPAG. PMID- 12095645 TI - Selective loss of 20S proteasome alpha-subunits in the substantia nigra pars compacta in Parkinson's disease. AB - The proteolytic activities of 26/20S proteasomes are impaired in the substantia nigra pars compacta (SNc) in sporadic Parkinson's disease (PD). In the present study, we examined the structural integrity of the proteasome by determining the levels of the beta- and alpha-subunits which together normally constitute the catalytic core of 26/20S proteasomes. Western blot analyzes and immunohistochemical staining revealed a major and selective loss of alpha subunits in dopaminergic neurons of the SNc but not in other brain regions in sporadic PD. This defect is known to cause the proteasome to become unstable and prevents its assembly with resultant impairment of enzymatic activity. Thus, structural and function defects in 26/20S proteasomes may underlie protein accumulation, formation of proteinaceous Lewy bodies and dopaminergic neuronal death in the SNc in sporadic PD. PMID- 12095646 TI - Age-related change of calbindin D-28k immunoreactive neurons in the rat main olfactory bulb. AB - We examined the aged-related changes of calbindin D-28k (CB)-immunoreactive (IR) neurons in the rat main olfactory bulb (MOB). The localization of CB-IR neurons was found to be almost entirely restricted to the glomerular layer. However, a few CB-IR neurons were observed in the granular layer. Prior to the 6th postnatal month (PM 6), the CB-IR neurons had long processes, and the population of CB-IR neurons had increased significantly compared to PM 12 and 24. At PM 12, CB-IR neurons showed a tendency to be smaller and have fewer dendrites than at guess. This phenomenon became remarkable at PM 24. The distinct reduction of CB-IR dendrites in glomeruli may be due to age-related functional restrictions. Increased calcium levels in the MOB neurons may be induced to allow cytotoxic event detection in the MOB neurons, and unbalanced Ca(2+)/Mg(2+)-ATPase may also induce aging-related MOB morphological changes. PMID- 12095647 TI - The involvement of 5-HT3 and 5-HT4 receptors in two models of gastrointestinal transit in mice. AB - Our aim was to study the involvement of 5-hydroxytryptamine (5-HT)(3) and 5-HT(4) receptors in two models of gastrointestinal transit (GIT) in mice: the 5 hydroxytryptophan (5-HTP)-induced diarrhea and intestinal inflammation produced by an irritant agent, croton oil (CO). 5-HTP (10 mg/kg) produced diarrhea that was significantly inhibited after pretreatment with ondansetron (5-HT(3) antagonist) or RS 39604 (5-HT(4) antagonist) (1-5 mg/kg). The GIT speed was increased after CO and 5-HTP administration. 5-HT(3-4) antagonists decreased GIT after 5-HTP-treatment but not after CO-treatment. Our results show that 5-HT(3) and 5-HT(4) receptors are involved in 5-HTP-induced diarrhea. This may be the reason why 5-HT(3-4) antagonists could be useful in the treatment of carcinoid syndrome diarrhea. 5-HT(3-4) antagonists were not effective in the modifications of GIT; nevertheless, they could be useful in the treatment of inflammatory bowel diseases because some symptoms as abdominal pain, discomfort or abnormal bowel function are modulated via 5-HT(3). PMID- 12095648 TI - Functional relation between caffeine- and muscarine-sensitive Ca2+ stores and no Ca2+ releasing action of cyclic adenosine diphosphate-ribose in guinea-pig adrenal chromaffin cells. AB - In voltage-clamped guinea-pig chromaffin cells, muscarine (50 microM) or caffeine (30 mM) produced a transient intracellular Ca(2+) concentration ([Ca(2+)](i)) increase, catecholamine release and an outward K(+) current mediated through Ca(2+) released from internal Ca(2+) stores at a holding potential of -40 mV. Caffeine followed by muscarine failed to evoke these responses, while muscarine followed by caffeine was effective in producing about 30% of [Ca(2+)](i) increase and catecholamine secretion. In cells dialyzed with inositol 1,4,5-trisphosphate (IP(3)), caffeine failed to produce the [Ca(2+)](i) increase. Intracellular application of cyclic adenosine 5'-diphosphate-ribose (cADP-ribose) or 8-bromo cADP-ribose exerted no effect on the resting [Ca(2+)](i) and the caffeine-induced [Ca(2+)](i) increase. These results suggest that IP(3)-sensitive stores are functionally divided into two subpopulations, sensitive and insensitive to caffeine, and it is unlikely that cADP-ribose plays a role as a Ca(2+) releaser in guinea-pig adrenal chromaffin cells. PMID- 12095649 TI - Association between intercellular adhesion molecule-1 E/K gene polymorphism and probable vascular dementia in humans. AB - Intercellular adhesion molecule-1 (ICAM-1) is implicated in the pathogenesis of ischemic cardiovascular disorders, including cerebral ischemia. A common polymorphism of the ICAM-1 gene (K469E) has been recently reported. In this case control study, we evaluated the association between this polymorphism and vascular dementia (VD) by studying 107 patients affected by probable VD and 115 age- and sex-matched controls. The frequency of the EE genotype was significantly higher in VD patients than controls (P=0.009). Logistic regression analysis indicated that the presence of the EE genotype significantly increased the risk of VD (odds ratio 3.25, P=0.024). Our findings support the hypothesis that ICAM-1 plays a role in the physiopathology of ischemic cerebrovascular disorders and suggest that genetic polymorphisms of ICAM-1 might be clinically important in the development and progression of neurodegenerative diseases. PMID- 12095650 TI - Coxsackievirus B4-induced neuronal apoptosis in rat cortical cultures. AB - Enterovirus infections of the central nervous system (CNS) are common and important causes of morbidity in immunocompromised children and adults. In this study we identify and characterize coxsackievirus B4-induced neuronal death. To investigate the CNS pathophysiology resulting from this viral infection, cultured rat neurons were infected with coxsackievirus B4 (CVB4) and nuclear morphology, phosphatidylserine (PS) externalization, and the effects of Actinomycin D or cycloheximide (CHX) were examined. CVB4 induced neuronal cell death within 24 h while PS externalization was apparent in cell bodies 16 h after CVB4 infection. Actinomycin D or CHX significantly reduced CVB4 induced-neuronal cell death in a dose-dependent manner. Pretreatment with CHX or actinomycin D also inhibited nuclear condensation, which occurred after CVB4 infection. However, the changes were relatively unresponsive to zVAD-fmk. These results suggest that CVB4 induces CHX- and actinomycin D-sensitive, but zVAD-fmk-insensitive neuronal apoptosis. PMID- 12095651 TI - Reduction of Alamar Blue, a novel redox indicator, is dependent on both the glycolytic and oxidative metabolism of glucose in rat cultured neurons. AB - Recently, it has been hypothesized that neurons mainly utilize lactate produced by astrocytes to generate energy (i.e. ATP). Regardless of numerous studies, whether lactate rather than glucose fuels neurons in vivo remains a matter of controversy. In the present study we have examined the substrate-preference of the reduction of Alamar Blue, a novel redox indicator, by cultured rat astroglia and neurons. Alamar Blue reduction is dependent on cellular reducing equivalents. Therefore, the substrate-preference of Alamar Blue reduction could reflect that for cellular energy production. Rates of Alamar Blue reduction by 2 mM glucose were two-fold higher in astroglia than those in neurons. Alamar Blue reduction induced by 2 mM lactate was 78% of that by 2 mM glucose in neurons while only 20% in astroglia. Nevertheless, glucose elicited larger Alamar Blue reduction than lactate both in astroglia and neurons. These results indicate that neuronal energy production in vivo depends on both the glycolytic and oxidative metabolism of glucose even though astroglia rely on glycolysis as has been reported. PMID- 12095652 TI - Ischemia-induced glutamate release in rat frontoparietal cortex after chronic alcohol and withdrawal. AB - High doses of ethanol increase stroke risk: in this context, a role for excitatory amino acids has been proposed. The present results show that, in frontoparietal cerebral cortex, chronic ethanol treatment (10% v/v in drinking water for 28 days) was able to slightly reduce glutamate release (evaluated through transdialysis coupled with high-pressure liquid chromatography) following focal ischemia as regards non-treated ischemic rats. This reduction was, however, not associated with decreased cerebral damage. In 24-h withdrawing rats, histological and morphometric analyzes showed an exacerbated cerebral damage coupled with higher glutamate and aspartate release compared to controls. These results suggest that adaptive changes following chronic ethanol consumption lead to an increased excitotoxicity that is particularly evident during the withdrawal condition. PMID- 12095653 TI - The Th1/E47cs-G apolipoprotein E (APOE) promoter allele is a risk factor for Alzheimer disease of very later onset. AB - Several polymorphisms in the apolipoprotein E (APOE) promoter region have been recently described. Of interest, APOE gene expression is increased in association with the -491AT polymorphism T-allele and decreased in relation to the Th1/E47cs polymorphism G-allele. In the present study we have investigated both polymorphisms in a series of 183 Alzheimer disease (AD) patients and 169 controls divided into age at onset/age dependent subgroups and the data obtained have been corrected for the presence of both expression-changing alleles in APOE homozygous individuals. Subsequently, the associations among APOE promoter polymorphisms, APOE4, and AD were assessed by chi-square and logistic regression analyses. Significantly, patients whose age at onset of AD was 80 years or more showed an association between the Th1/E47cs-G allele and AD that was independent of the APOE4 allele. PMID- 12095654 TI - Myenteric neurons and interstitial cells of Cajal of mouse colon express several nitric oxide synthase isoforms. AB - Information on equipment and subcellular distribution of nitric oxide synthase (NOS) isoforms in myenteric neurons and pacemaker cells (ICC) might help to identify nitric oxide (NO) pathway(s) acting on gastrointestinal motility. In sections of mouse colon labelled with neuronal (n)NOS, endothelial (e)NOS and inducible (i)NOS antibodies, all myenteric neurons co-expressed eNOS and iNOS and a subpopulation of them co-expressed nNOS. ICC co-expressed nNOS and eNOS. In the neurons, nNOS-labeling was intracytoplasmatic, in the ICC at cell periphery. In both cell types, eNOS-labeling was on intracytoplasmatic granules, likely mitochondria. In conclusion, myenteric neurons and ICC co-express several NOS isoforms with specific subcellular distribution. Different nNOS splice variants are presumably present: intracytoplasmatic nNOSbeta and nNOSalpha producing neurogenic NO, plasma membrane-bound nNOSalpha producing ICCgenic NO. eNOS might be implicated in mitochondrial respiration and, in ICC, also in pacemaker activity. Neurons express iNOS also in basal condition. PMID- 12095655 TI - The cannabinoids R(-)-7-hydroxy-delta-6-tetra-hydrocannabinol-dimethylheptyl (HU 210), 2-O-arachidonoylglycerylether (HU-310) and arachidonyl-2-chloroethylamide (ACEA) increase isoflurane provoked sleep duration by activation of cannabinoids 1 (CB1)-receptors in mice. AB - Cannabinoids produce antinociception via specific cannabinoid receptor activation, but there are also non-receptor mediated effects like for example the activation of the arachidonic acid cascade. Here we investigate the influence of cannabinoids (CB) on sleep duration after isoflurane anesthesia. We found that the CB receptor agonists R(-)-7-hydroxy-delta-6-tetra-hydrocannabinol dimethylheptyl (HU-210) (0.1 mg/kg), 2-O-arachidonoylglycerylether (30 mg/kg) and arachidonyl-2-chloroethylamide (3 mg/kg) significantly prolong the duration of isoflurane induced sleep in mice (P<0.05). This effect was absent when co injecting the selective CB(1) antagonist N-(piperidin-1-yl)-5-(4-iodophenyl)-1 (2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (1 mg/kg). Furthermore, HU-210 was ineffective in CB(1) receptor knockout mice (CB(1)-/-). Our behavioral tests (tail flick, rotarod) indicate that the sleep latency can be prolonged even at low drug dosages which do not influence thermal nociception. In the chosen dosages thimerosal (20 mg/kg), 2-AG (10 mg/kg), R(1)-methanandamide (R(1)-MAEA) (10 mg/kg) and flurbiprofen (27 mg/kg) were ineffective to increase sleep duration. PMID- 12095656 TI - Cyclic guanosine 5' monophosphate (GMP) prevents expression of neuronal nitric oxide synthase and apoptosis in motor neurons deprived of trophic factors in rats. AB - Deprivation of trophic factors induces expression of neuronal nitric oxide synthase (NOS) and nitric oxide production in cultured motor neurons, leading to apoptosis. Motor neuron apoptosis requires the simultaneous production of nitric oxide and superoxide and is associated with increased nitrotyrosine immunoreactivity. Nitric oxide also stimulates cyclic guanosine 5' monophosphate (cGMP) synthesis, which enhances the survival of motor neurons treated with brain derived trophic factor (BDNF). Here we report that cGMP analogs blocked neuronal NOS induction, nitrotyrosine accumulation, and prevented apoptosis for up to 3 day of motor neurons deprived of trophic factors. Low concentrations of exogenous nitric oxide (<100 nM), which are not toxic for BDNF-treated cultures, reversed the protective effect of cGMP. These results suggest that elevation of cGMP could decrease nitric oxide production, and thereby preventing motor neuron apoptosis. PMID- 12095657 TI - Dexamethasone induces dendritic alteration but not apoptosis in the neurons of the hippocampus in postnatal rats. AB - One day old rats received daily injections of dexamethasone and were sacrificed 24 h after the 1st and 7th injections. Neuronal death by apoptosis in the hippocampus was investigated by immunohistochemistry using bcl2, bax and caspase3 antibodies. The immunoreactivity expressed by the pyramidal neurons and the dentate granule cells with these antibodies was comparable in the dexamethasone treated and control rats injected with saline. At the ultrastructural level, the dendrites showed vacuolation indicative of degeneration in the dexamethasone administered rats. Results of reverse transcriptase-polymerase chain reaction analysis showed that bcl2 and bax mRNA was constitutively expressed in the hippocampus of control rats and showed no significant change in the dexamethasone treated rats. The results of this study indicate that dexamethasone induces degeneration of the dendrites but does not induce neuronal apoptosis in the hippocampus of postnatal rats. PMID- 12095658 TI - Immunohistochemical study of synphilin-1 in brains of patients with dementia with Lewy bodies - synphilin-1 is non-specifically implicated in the formation of different neuronal cytoskeletal inclusions. AB - Dementia with Lewy bodies brains were immunohistochemically investigated using anti-synphilin-1 antibodies. The alpha-synuclein-positive brainstem type and well defined cortical type Lewy bodies (LB) were positive for synphilin-1, while ill defined LB and LB-related neurites were negative, suggesting that synphilin-1 does not directly associate with alpha-synuclein. Synphilin-1-positive LB were double-positive for phosphorylated neurofilament. In addition, tau-positive neurofibrillary tangles (NFT) were positive for synphilin-1, while neuropil threads were negative. Immunoelectron microscopically, synphilin-1 was located on filamentous components in cortical type LB and on paired helical filaments in NFT. It is likely that synphilin-1 accumulates in the cell body according to the axonal transport blockage, and associates with abnormal cytoskeltons during the formation of LB or NFT, suggesting that synphilin-1 is non-specifically implicated in the formation of different neuronal cytoskeletal inclusions. PMID- 12095659 TI - Effects of diltiazem and MK-801 on morphine analgesia and pharmacokinetics in mice. AB - The effects of diltiazem, an L-type calcium channel blocker, and MK-801, a non competitive N-methyl-D-aspartate (NMDA) receptor antagonist on morphine analgesia and pharmacokinetics were examined in mice. Mice received a subcutaneous injection of morphine (3.2 mg/kg) 30 min after a subcutaneous injection of diltiazem or MK-801. Diltiazem (20-60 mg/kg) potentiated morphine analgesia and increased serum morphine levels in a dose-dependent manner. MK-801 (0.3 mg/kg) significantly attenuated morphine analgesia but had no significant effect on serum or brain morphine levels. These results suggest that a modification of morphine metabolism is involved, at least in part, in the ability of diltiazem to enhance morphine analgesia, whereas MK-801 attenuates morphine analgesia without affecting morphine pharmacokinetics. PMID- 12095660 TI - 4-MTA: a new synthetic drug on the dance scene. AB - 4-MTA (p-methylthioamphetamine) is a new synthetic sulphur derivative of amphetamine that has been associated with six deaths since it was first identified in Europe in 1997. Sold as 'ecstasy' or 'Flatliners', the drug like MDMA is a potent serotonin releaser. Using a self-nominated sampling methodology we accessed over 1000 dance drug users through a magazine survey. Ten percent of respondents thought that they had used 4-MTA. Those with experience of 4-MTA tended to come from a sub-population of heavy 'ecstasy'-users. Responses to the effects of the drug were mixed, although about a quarter of those who believed that they had tried 4-MTA thought that they would use it again. PMID- 12095661 TI - Treating hepatitis C in methadone maintenance patients: an interim analysis. AB - OBJECTIVE: This study evaluates the safety, tolerability, and efficacy of interferon/ribavirin combination therapy in methadone maintenance (MM) patients with active hepatitis C (HCV). End-of-treatment results are presented as an interim analysis of efficacy. METHODS: Fifty eligible MM patients with active HCV and concomitant liver fibrosis were treated with interferon/ribavirin combination therapy using standard dosing regimens. Patients with active drug or alcohol use at the start of treatment, severe or untreated psychiatric illness, and/or decompensated liver disease were excluded. RESULTS: Treated MM patients were older, had a longer history of HCV infection, a high prevalence of psychiatric illness, and had substantially more liver fibrosis than in previous studies of HCV treatment in non-opioid-dependent populations. Seventy-eight percent completed the 6-12 month course of treatment. The end-of-treatment virologic response rate was 64% in patients completing treatment and 54% on an intent-to treat basis. CONCLUSION: Methadone maintenance patients exhibit a number of factors that make HCV treatment more difficult: they are older, have a higher prevalence of psychiatric illness, and show evidence of more advanced liver disease. Despite this, their end-of-treatment response rate to interferon/ribavirin combination therapy is similar to that of patients without a history of IDU. Further analysis of the sustained virologic response (SVR) rate is planned at the study's completion. These preliminary results show that MM patients are in need of timely HCV diagnosis, and should be considered good candidates for referral and HCV treatment. PMID- 12095662 TI - Abstinence reinforcement maintenance contingency and one-year follow-up. AB - BACKGROUND: Relapse to drug use is often seen when contingencies designed to reduce drug use are discontinued. This paper reports on a stepdown maintenance contingency and 1-year follow-up in 110 patients who were maintained on methadone (50 or 70 mg/day) and who had completed a contingency management trial targeted to decreasing their opiate use. In the prior study (induction phase, 8 weeks) participants received vouchers for each opiate-negative urine screen or noncontingently. METHODS: In this study (maintenance phase, 12 weeks), participants were rerandomized to receive vouchers and take-home methadone doses contingent on providing opiate-negative urine specimens (N=55) or noncontingently (N=55). Since participants had been rerandomized from induction-phase contingencies, most study data were analyzed as if from a 2 x 2 (inductionxmaintenance) design. Follow-up interviews were conducted at 3, 6, and 12 months after study participation. RESULTS: Patients who received the maintenance contingency following an 8-week induction contingency had better outcomes than those who received noncontingent incentives in either the maintenance or induction phases of the trial. Good outcome at follow-up was predicted by enrollment in methadone maintenance after the study. Significantly more participants in the maintenance contingency group transferred directly to another methadone program. CONCLUSION: These findings support the therapeutic value of extending the duration of contingency management and long-term methadone maintenance. PMID- 12095663 TI - Cognitive functioning among infants of alcoholic fathers. AB - Although there is considerable evidence that suggests that children of alcoholic fathers are at risk for maladaptive developmental outcomes, there is a large amount of heterogeneity in these outcomes. This study examined this hypothesis using broad measures of mental and language development during the toddler period as a function of fathers' alcoholism and associated risk factors. Participants included 102 families in which the father was alcoholic, 20 families in which the father was alcoholic and the mother was a heavy drinker and 104 control families matched in terms of maternal education, race/ethnicity, child gender, marital status and number of children. Families, in which the mother engaged in significant prenatal drinking or drug use, were excluded. Infants were tested at 12, 18 and 24 months old. Analyses revealed no significant differences between controls and infants of severe alcoholics, antisocial alcoholics, depressed alcoholics or family history positive alcoholics. There were also no differences between controls and infants in families with an alcoholic father/heavy drinking mother. The results suggest that overall mental development was not associated with paternal alcoholism. This raises the possibility that later problems in these areas may reflect more specific cognitive processes or the impact of parenting and behavioral problems among these children. PMID- 12095665 TI - Acute effects of oral cocaine on inhibitory control of behavior in humans. AB - Studies of humans show that individuals with histories of cocaine abuse display reduced inhibitory control over behavioral impulses. The present study tested the effects of oral cocaine on the ability to inhibit behavior in humans. Eight adult volunteers (seven men and one woman) with a history of cocaine abuse participated as in-patient volunteers. Response inhibition and response execution were measured by a stop-signal paradigm using a choice reaction time task that engaged subjects in responding to go-signals when stop-signals occasionally informed them to inhibit the response. Subjects' performance on the task was tested just before and 1 h after a randomized, double-blind administration of 0 mg (placebo), 50, 100, and 150 mg of oral cocaine HCl. Cocaine reduced subjects' ability to inhibit responses to stop-signals. By contrast, no effect of cocaine was observed on the ability to execute responses in terms of their speed and accuracy. Subjective and physiological effects of cocaine were also observed. Together, the findings indicate that acute administration of cocaine can impair the ability to inhibit behavioral responses at doses that do not affect the ability to respond. These findings are important because they identify a specific disinhibiting effect of cocaine that could help explain the documented association between long-term cocaine use and poor impulse control. PMID- 12095664 TI - Comparison of the subjective, physiological, and psychomotor effects of atomoxetine and methylphenidate in light drug users. AB - This study compared the subjective, physiological, and psychomotor effects of atomoxetine and methylphenidate with placebo in healthy volunteers. Sixteen non dependent light drug users participated in six experimental sessions, receiving placebo, atomoxetine (20, 45 and 90 mg) and methylphenidate (20 and 40 mg) using a double-blind, Latin square design. Subjective drug effects were assessed using Visual Analog Scales (VAS), the Addiction Research Center Inventory (ARCI) and Adjective Rating Scales (ARS). Psychomotor performance was evaluated using the Digit Symbol Substitution Test (DSST). Physiological measures were also collected throughout the sessions. Assessments were conducted before drug administration and 30, 60, 90, 120, 150, 180 and 240 min following dosing. Forty milligrams methylphenidate produced significant increases on the stimulant portions of the VAS and ARS and the benzedrine, amphetamine, morphine-benzedrine and lysergic acid diethylamine (LSD) subscales of the ARCI relative to placebo. Ninety mg atomoxetine was reported to be unpleasurable relative to placebo as indicated by significant increases on the 'bad' and 'sick' portions of the VAS, and on the LSD subscale of the ARCI. Compared with placebo, both methylphenidate doses significantly increased systolic blood pressure (BP) and heart rate (HR). For atomoxetine, 90 mg increased diastolic BP, 45 and 90 mg increased systolic BP, and all three doses increased HR relative to placebo. Neither compound produced significant differences from placebo on DSST performance. These results suggest that atomoxetine does not induce subjective effects similar to methylphenidate and suggest that it is unlikely that atomoxetine will have abuse liability. PMID- 12095666 TI - Prison based detoxification for opioid dependence: a randomised double blind controlled trial of lofexidine and methadone. AB - This paper reports results from the first controlled trial of opioid withdrawal treatment in the UK using lofexidine in a prison setting. Seventy-four opioid dependent male inmates at a Southern England prison were randomised to receive either methadone (the standard prison treatment) or lofexidine using a randomised double-blind design. No significant statistical difference between the treatment groups was found in relation to the primary variable of severity of withdrawal symptoms (effect size=0.12). No discernible difference was found in the sitting blood pressure or heart rate of the two groups during the trial. These results provide support for the use of lofexidine for the management of opioid detoxification in the prison setting. PMID- 12095667 TI - A randomized, double-blind, placebo-controlled safety study of high-dose dextromethorphan in methadone-maintained male inpatients. AB - The NMDA antagonist dextromethorphan hydrobromide (DM) may be useful in the treatment of opioid dependence, particularly as a means of reducing tolerance to methadone during replacement therapy. As a prelude to clinical efficacy studies, a randomized, double-blind, placebo-controlled study examined the safety of DM in combination with methadone in inpatient, opiate-dependent volunteers. Male participants received daily methadone (50-70 mg/day) and either DM (n=10) or placebo (n=5) during the 12-day active medication phase of the study. DM participants received doses of 120, 240, and 480 mg/day in increasing order (4 days each). DM at high doses caused mild elevations of heart rate, blood pressure, temperature, and plasma bromide. However, none of these effects was clinically significant. DM caused no significant changes in respiration, pupil diameter, or subjective drug effects measured by standard scales. Participants in the DM group reported many more adverse events than did subjects on placebo (173 vs. 21), but these effects were not clinically serious. The most commonly reported side effects were sleepiness and drowsiness. Several participants reported intoxicating effects at the highest dose. Overall, DM was well-tolerated by the methadone-maintained opiate-dependent subjects studied here. These results support the further exploration of DM as an adjunct medication during methadone replacement therapy. PMID- 12095669 TI - Decision biases and persistent illicit drug use: an experimental study of distributed choice and addiction. AB - This experiment tested the hypothesis that differences in drug use are correlated with differences in decision making. The subjects were 22 drug clinic patients who had used either opiates or stimulants for an average of 10 years, and 21 community residents who reported that they had rarely used illicit addictive drugs. The procedure consisted of a series of binary choices with two consequences; they earned money and determined the intervals that separated choice trials. Each choice earned the same amount of money, but one initiated a shorter delay to the next trial, whereas the other initiated a shorter delay as averaged over the next two trials. Shorter delays were advantageous in that they increased the overall rate of earnings and they reduced the time spent waiting for the next trial. Thus, one choice was better from the perspective of the current trial, while the other choice was better from the perspective of two or more consecutive trials. Drug-clinic patients were more likely to favor the one trial solution compared with control subjects, who were more likely to favor the two-trial solution. There were five different choice games, with different versions varying in the magnitude of the advantage for switching from the two trial to the one-trial solution. Drug clinic and control subjects differed most in the games in which the immediate advantage of the one-trial solution was larger, and all subjects were more likely to choose the global solution when the incentive for switching to the one-trial solution was lower. The results support the view that individual differences in decision making influence the course of illicit drug use. PMID- 12095668 TI - Comparing attentional bias to smoking cues in current smokers, former smokers, and non-smokers using a dot-probe task. AB - Much evidence documents that individuals with emotional and drug-use disorders demonstrate biased attention toward stimuli associated with their disorder. This bias appears to diminish following successful treatment. Two studies examined whether current cigarette smokers show biased attention toward smoking-related images compared with non-smokers (Studies 1 and 2) and whether this bias is less pronounced in former smokers (Study 2). Attentional bias toward cigarette-related photographs was examined using the dot-probe task. Pairs of images (one smoking related) appeared side by side for 500 ms on a computer screen prior to the presentation of a probe (an asterisk) replacing one of the photographs. Subjects struck a key as quickly as possible to indicate the probe location. Attentional bias was defined as faster reaction times when the probe replaced the smoking related image. In both studies, current smokers displayed significantly greater attentional bias toward cigarette stimuli than did non-smokers. Former smokers in Study 2 displayed an intermediate level of bias, but did not differ significantly in bias score from either of the other groups. These results support further use of the dot-probe task as a measure of attentional bias in non-abstinent smokers and in individuals undergoing smoking cessation treatment. PMID- 12095670 TI - 'Diagnostic orphans' among young adult cannabis users: persons who report dependence symptoms but do not meet diagnostic criteria. AB - OBJECTIVE: To examine the characteristics of 'diagnostic orphans' among cannabis users-those who report one or two symptoms of DSM-IV dependence but do not meet diagnostic criteria for DSM-IV abuse or dependence. METHOD: Data were collected from a representative population cohort of 1601 young adults aged 20-21 years. Those who reported that they had used cannabis at least weekly at some point within the past year were assessed for symptoms of DSM-IV cannabis abuse and dependence using the Composite International Diagnostic Interview. RESULTS: Approximately 2.8% of the cohort could be classified as diagnostic orphans, with another 3.0 and 7.5% meeting criteria for abuse and dependence, respectively. Diagnostic orphans were: similar to those who met criteria for cannabis abuse or dependence in terms of demographic characteristics; similar to those who met criteria for cannabis abuse in terms of cannabis use patterns; and similar to those who met criteria for abuse and dependence in their rates of heavy alcohol use and DSM-IV alcohol dependence. However, they did not appear to have elevated rates of illicit drug use or mental health problems compared to non users. CONCLUSIONS: Diagnostic orphans reported using cannabis in a manner similar to persons meeting criteria for cannabis abuse, and had similar rates of alcohol dependence and other illicit drug use. Strict adherence to DSM-IV diagnoses of abuse and dependence may overlook a substantial proportion of young persons who experience cannabis-related problems. There is a need to consider (a) subthreshold levels of cannabis-related problems among those seeking treatment for other problems; and (b) interventions for this group to prevent escalation of such problems. PMID- 12095671 TI - Bystander resuscitation attempts at heroin overdose: does it improve outcomes? AB - STUDY OBJECTIVE: To document the characteristics and effectiveness of cardiopulmonary resuscitation (CPR) at non-fatal heroin overdose events in Melbourne, Australia. METHODS: A retrospective analysis of a computerised database of ambulance attendance records at non-fatal heroin overdose cases for the period 1/12/1998 to 31/7/2000 was undertaken. MAIN OUTCOME MEASURES: The main outcome measure was the rate of patient hospitalisation. The rate of CPR administration at heroin overdose cases was also examined, along with characteristics of the attendance, such as the age and sex of the overdose case, the relationship of person providing CPR to the overdose case as well as the location, time and date of the event. RESULTS: CPR was administered prior to ambulance arrival in 579 heroin overdose cases (9.4% of total heroin overdose cases attended) between 1/12/98 and 31/7/2000. A greater proportion of female overdose cases were administered CPR than males and CPR administrations were evenly distributed across attendances occurring in private and public locations. Bystander administration of CPR prior to ambulance attendance resulted in a significantly lower rate of heroin user hospitalisation (14.5%) compared to cases where bystander CPR was not administered (18.8%). CONCLUSIONS: While CPR administration prior to ambulance attendance at heroin overdose events is relatively uncommon (especially compared to out-of-hospital cardiac arrest), such administration was associated with a statistically significant improvement in clinical outcomes in cases of non-fatal heroin overdose. These findings suggest that the provision of CPR training to people likely to come into contact with heroin overdose events may be an effective strategy at minimising consequent overdose-related harm. PMID- 12095672 TI - Mediating mechanisms for the impact of bupropion in smoking cessation treatment. AB - Several studies have documented the effectiveness of bupropion for smoking cessation, yet little is known about the mechanisms by which it facilitates abstinence. In this placebo-controlled randomized trial. We examined whether bupropion's effects on cessation were mediated by changes in withdrawal and/or negative or positive affect (PA). Two hundred and fifty-one smokers received 10 week treatment with bupropion or placebo, plus behavioral counseling. Changes in affect and withdrawal symptoms from pre-quit to 1 week post quit were examined as mediating variables in structural equation models. Cotinine-verified 7-day point prevalence cessation rates at the end of treatment (8-weeks post quit date) were 48% for bupropion and 29% for placebo (P=0.001). There were significant treatment effects on withdrawal and negative affect (NA); however, only change in NA predicted cessation. In a path model, change in NA was a significant mediator of bupropion's effects on cessation. However, the proportion of variance accounted for by this mediator was small, suggesting that other unmeasured factors play an important role. Laboratory-based paradigms may be useful to identify other mediators of bupropion's effects, thereby pointing to mechanisms of effect that can be bolstered in future treatment studies. PMID- 12095673 TI - Emx2: a gene responsible for cortical development, regionalization and area specification. AB - Mouse Emx2 homeobox gene is a very good dorsal marker for the developing cerebral cortex, as it is mainly expressed in this area from the very beginning of corticogenesis. Its cortical expression includes proliferating neuroblasts of the neuroepithelium, or ventricular zone, and the postmitotic Cajal-Retzius cells, known to control neuronal radial migration. Analysis of the phenotype of Emx2 null embryos has shown that this transcription factor plays important roles in neuroblast proliferation, migration and differentiation, as well as in the development of the diencephalon, where it has been shown to cooperate with Otx2. Moreover, the graded distribution of EMX2 homeoprotein along the antero-posterior and medial-lateral cortical axis, is responsible for the patterning of the forebrain, in particular for the specification process that defines cortical territories and area identity during neocortical development. Emx2 participates to this process as Pax6 and COUP-TF1. Finally Emx2 is very interesting from the evolutionary point of view, as it has been shown to share a high degree of homology in its sequence and function with Vax1, another homeobox gene regulating basal forebrain development. This homology traces back to Emx and Vax gene families, which are strongly related, as they are thought to derive from a common ancestor gene. PMID- 12095674 TI - Genomic organization and chromosomal localization of the mouse protein kinase C alpha gene. AB - Protein kinase C (PKC) is a family of ten isoforms of phospholipid-dependent serine/threonine kinases, which participate in many cellular responses including cell growth, differentiation, and tumorigenesis. Of the isoforms, PKC alpha is distributed ubiquitously in almost all tissues and involved in various signal transductions. Furthermore, PKC alpha plays an important role in the growth and malignant progression of some tumor cell lines. Elucidating the roles of PKC alpha in vivo would lead to understanding of the mechanism of tumorigenesis and other biological functions. In this study, we isolated and characterized genomic DNA clones of the mouse PKC alpha gene (Prkca). The Prkca gene was a unigene consisting of 17 exons and spanning at least 116 kb. All the exon-intron boundaries followed the GT/AG rule. The genomic structure of PKC alpha was markedly conserved among the mouse, human, and fly. By radiation hybrid mapping, the Prkca gene was closely linked to sequence-tagged site marker D11Mit258 that locates 65.0 cM from the centromere of chromosome 11, and its transcription was towards the centromere. This study shows that generation of PKC alpha-mutant mice may reveal the PKC alpha function in vivo. PMID- 12095675 TI - Characterization and tissue-specific expression of human GSK-3-binding proteins FRAT1 and FRAT2. AB - We have isolated the entire coding sequence of human FRAT2 (frequently rearranged in advanced T-cell lymphomas-2). It exhibits appreciable amino acid identity to FRAT1 (77%) which was initially isolated as frequently being overexpressed in a murine leukemia virus insertion model in murine tumors. FRAT proteins are thought to play a role in Wnt signaling. They can bind to glycogen synthase kinase-3 (GSK 3) and Dishevelled, two proteins involved in Wnt signal transduction. Both hFRAT1 and hFRAT2 are intronless genes localized to the same portion of chromosome 10q24.1 and separated by only 10.7 kb. In a broad range of human tissues FRAT1 and FRAT2 are readily detected and expressed in a near identical pattern. Both species are repressed when the human embryonal carcinoma cell line, NT2/D1, is induced to differentiate with all-trans retinoic acid (RA). This treatment had no appreciable effect on FRAT levels in two other RA-sensitive cell lines that were not of germ cell tumor origin. The overlapping expression patterns suggest these two genes share a regulatory region. Both FRAT genes exhibited three species of mRNA, which varied in representation between tissues. When transiently overexpressed in COS-1 cells, the FRAT proteins were detected in the cytosol and concentrated in the nucleus. Both hFRAT1 and hFRAT2 are implicated in the selective modulation of GSK-3 activity via the Wnt signaling pathway. This study provides a foundation from which to examine the role these proteins play in Wnt dependent and -independent processes. PMID- 12095676 TI - Isolation, characterization and mapping of the mouse and human RB1CC1 genes. AB - RB1CC1 (RB1-inducible Coiled-Coil 1), a putative transcription factor implicated in the regulation of RB1 (retinoblastoma 1) expression, was recently identified in a screen for genes involved in multi-drug resistance to anticancer agents. Information about the RB1CC1 gene is limited, however, and its biological function is not determined. Here we report the isolation, characterization and mapping of the mouse RB1CC1 gene (Rb1cc1), together with further characterization of the human RB1CC1 gene. Mouse Rb1cc1 encodes 1588 amino acids, sharing 89% identity and key sequence motifs with its human counterpart. Rb1cc1 is expressed abundantly in heart and testis, with lower levels detected in lung and spleen. Immunohistochemical analysis revealed the Rb1cc1 and Rb1 proteins are co localized in the cell nuclei of NIH3T3-3 cell and various mouse tissues. The human and mouse RB1CC1 genes, both of which contain 24 exons, span 74 kb on chromosome 8q11.2 and 57 kb on chromosome 1A2-4, respectively. Conserved sequence motifs and nuclear localization suggest that the RB1CC1 proteins function as transcription factors. PMID- 12095677 TI - The murine pantothenate kinase (Pank1) gene encodes two differentially regulated pantothenate kinase isozymes. AB - Pantothenate kinase (PanK) is a rate-determining enzyme in coenzyme A (CoA) biosynthesis. The mouse murine pantothenate kinase (Pank1) gene consists of seven introns and eight exons and is located on chromosome 19 (19C2-3). Two biochemically distinct PanK1 protein isoforms, PanK1 alpha and PanK1 beta, are encoded by the Pank1 gene. Both proteins have the same 363 amino acid catalytic domain encoded by exons 2 through 7. The PanK1 beta transcript begins with exon 1 beta and translates into a ten-residue amino terminus plus the catalytic domain. The PanK1 alpha transcript initiates at an alternate upstream site at exon 1 alpha which is spliced with exon 2, excluding exon 1 beta. Exon 1 alpha encodes a 184-residue regulatory domain at the amino terminus of the PanK1 alpha protein that confers feedback inhibition by free CoA and long-chain acyl-CoA, and increases the regulation of PanK enzyme activity by acetyl-CoA and malonyl-CoA. Differential expression of the PanK1 alpha and PanK1 beta transcripts would alter the amount of CoA produced in cells as a function of the ratio of free CoA to acetyl-CoA, a reflection of the metabolic status of the tissue. PMID- 12095679 TI - Fructose-1,6-bisphosphatase genes in animals. AB - A comparison of the amino acid sequences of the liver and muscle fructose-1,6 bisphosphatase (FbPase) isoforms in primates and rodents suggested an ancient duplication event leading to the corresponding genes. We investigated the presence of both genes in the rabbit (order lagomorphs) and in species belonging to further distantly related metazoan taxa. By an analysis of the available complete genomes and proteomes of the nematode Caenorhabditis elegans and of Drosophila melanogaster only one sequence homologous to known FbPases was found in each species. The corresponding mRNAs were characterized by cDNA sequencing. We then carried out reverse transcription-polymerase chain reactions to amplify central fragments of the FbPase cDNAs from liver and muscle of Gallus gallus, Xenopus laevis, and Esox lucius, respectively. Their sequencing revealed that (i) the livers of chicken, frog, and fish contain mRNAs which are closely related to mammalian liver FbPase mRNAs, (ii) chicken muscle contains an mRNA which is most homologous to mammalian muscle FbPase mRNAs, (iii) frog muscle contains both a liver-type and a muscle-type FbPase mRNA, while (iv) in fish muscle no FbPase mRNA could be detected by our approach despite the doubtless presence of the enzyme in this organ. An alignment of the partial amino acid sequences of the different FbPases showed that the residues that are thought to be in contact with the substrate, fructose-2,6-bisphosphate, and Mg(2+) are totally conserved, while some amino acids having contact with adenosine monophosphate were found to vary among several species. The question of what might be the advantage of having more than one gene coding for FbPase per haploid genome is discussed. PMID- 12095678 TI - Cloning and characterization of a levanbiohydrolase from Microbacterium laevaniformans ATCC 15953. AB - An extracellular levanbiohydrolase gene, levM, from Microbacterium laevaniformans ATCC 15953 was cloned and its nucleotide sequence was determined. Nucleotide sequence analysis of this gene revealed a 1863 bp open reading frame coding for a protein of 621 amino acids. The deduced amino acid sequence of the levM gene exhibited 28-47% sequence identities with levanases, levanfructotransferases, and inulinases. The LevM was overexpressed by using a T7 promoter in Escherichia coli BL21 (DE3) and purified 24-fold from culture supernatant. The molecular weight of this enzyme was 68,800 Da based on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimum pH and temperature of this enzyme for levan degradation was pH 6.0 and 30 degrees C, respectively. Thin-layer and high performance liquid chromatography analyses proved that the enzyme produced mostly levanbiose from levan in an exo-acting manner. The recombinant enzyme also hydrolyzed inulin, 1-kestose, and nystose, indicating that the enzyme cleaves not only beta-2,6-linkage of levan but also beta-2,1-linkage of fructooligosaccharides. This is the first report on a gene encoding a levanbiohydrolase that produces levanbiose as a major degradation product. PMID- 12095680 TI - The ADAM1a and ADAM1b genes, instead of the ADAM1 (fertilin alpha) gene, are localized on mouse chromosome 5. AB - Fertilin is reported to be a heterodimeric protein composed of A Disintegrin And Metalloprotease 1 (ADAM1, fertilin alpha) and ADAM2 (fertilin beta) located on the sperm surface. In the process of clarifying the molecular basis of mouse ADAM1, we have identified two intron-less mouse genes encoding different isoforms of ADAM1, termed ADAM1a and ADAM1b. The amino acid sequences of ADAM1a and ADAM1b deduced from the DNA sequences were homologous to each other (99% identity) in the pro- and metalloprotease domains, whereas the C-terminal half region of ADAM1a, including the disintegrin and Cys-rich domains, shared only a low degree of identity (37%) with that of ADAM1b. These two genes were both localized on mouse chromosome 5 as a single copy gene, and were expressed specifically in the testis. These data demonstrate the presence of the ADAM1a (Adam1a) and ADAM1b (Adam1b) genes in mouse, instead of the ADAM1 gene, and may imply different roles of ADAM1a and ADAM1b in spermatogenesis, sperm maturation, and/or fertilization. PMID- 12095681 TI - Cloning and characterization of a gene (avaA) from Aspergillus nidulans encoding a small GTPase involved in vacuolar biogenesis. AB - Several genes that play roles in vacuolar biogenesis and targeting of proteins to vacuoles have been characterized in Saccharomyces cerevisiae. Although the vacuole is one of the most prominent compartments, little is known about molecular mechanism of vacuolar biogenesis in filamentous fungi. Vam4/Ypt7p, a small GTPase of the Rab/Ypt family in S. cerevisiae, plays a vital role in homotypic vacuole fusion. We describe the isolation of the avaA gene from Aspergillus nidulans as a homologue of the VAM4/YPT7. Predicted 205 amino acids protein encoded by the avaA showed 67 and 72% identity with yeast Vam4/Ypt7p and human Rab7, respectively. The avaA disruptants exhibited highly fragmented vacuoles. We introduced mutations into the avaA gene, which alter nucleotide binding characteristics of the gene products. Replacement of the avaA gene to the GDP-bound form mutant resulted in fragmentation of vacuoles. Overexpression of the GTP-bound form of avaA severely inhibited the hyphal growth and caused abnormal swelling of vacuoles. These results suggest that A. nidulans AvaA functions in the vacuolar biogenesis. PMID- 12095682 TI - Isolation of Drosophila activin and follistatin cDNAs using novel MACH amplification protocols. AB - With the genomic sequence of multicellular organisms such as Caenorhabditis elegans, Drosophila melanogaster, and Homo sapiens completed and others to be finished in the near future, the focus has shifted from accumulating sequence information to the prediction and analysis of genes within the completed genomes. Unfortunately, presently available computer programs do not always accurately predict gene structure such as mRNA and translation start sites or intron/exon boundaries. The only way to be certain about a gene's structure is to isolate and characterize its cDNA. Since the screening of libraries is a time-consuming, labor-intensive process that sometimes fails to yield the desired clone, we searched for faster, more efficient ways to isolate cDNAs. In this study, we describe two methods for amplification and isolation of cDNAs from plasmid libraries that requires no hybridization (MACH). With the polymerase chain reaction-based MACH-2 protocol, we present a strategy that requires little DNA sequence information to selectively isolate the longest cDNA variant from plasmid libraries in about 3 days. Our protocols were used to isolate cDNAs for the Drosophila activin and follistatin genes. PMID- 12095683 TI - Identification of putative interaction partners for the Xenopus Polycomb-group protein Xeed. AB - The extra sex combs (esc) gene of Drosophila and its mammalian homologue embryonic ectoderm development (eed) play pivotal roles in establishing Polycomb group (Pc-G) mediated transcriptional silencing of regulatory genes during early development. We have carried out a two-hybrid screen in yeast to identify maternally expressed proteins that interact directly with the product of the Xenopus eed homologue, Xeed. Xeed-interacting proteins that were recovered in this screen included a maternal Xenopus histone deacetylase (HDACm), the Xeed protein itself, and a Xenopus homologue of Enhancer of zeste (XEZ) - a second member of the Pc-G that is closely related by sequence similarity to histone methyltransferases. We have also identified a novel interaction between Xeed and a component of the Xenopus basal transcription machinery, TAF(II)32. We show for the first time that each of these proteins interacts with the Xeed polypeptide, both in the yeast two-hybrid assay and in vitro using purified recombinant proteins. XEZ, HDACm and TAF(II)32 mRNAs are all strongly co-expressed with Xeed mRNA in the fertilized egg, further suggesting that their encoded proteins could interact with Xeed during early embryonic development. Our observations support a multi-step model for the onset of transcriptional silencing in which Xeed binds to and inhibits the function of the transcription initiation complex and also recruits proteins that mediate the acquisition by associated chromatin of epigenetically heritable, post-translational modifications. PMID- 12095684 TI - Analysis of transcriptomes of human malaria parasite Plasmodium falciparum using full-length enriched library: identification of novel genes and diverse transcription start sites of messenger RNAs. AB - Now that the sequencing of the complete genome of the human malaria parasite Plasmodium falciparum is now underway, importance of analyses of complementary DNAs (cDNAs) is looming up. We constructed a full-length-enriched cDNA library from erythrocytic stage P. falciparum using the 'oligo-capping' method (Nucleic Acids Res. 29 (2001) 70). In this report we describe the novel genes identified using this library and detailed characterization of transcriptional start site of knob-associated histidine rich protein gene. Contrary to the previous report we conclude all the transcripts of plasmodium genes have diverse start sites. Sequence comparisons between the cDNAs and the complete sequences of chromosomes 2 identified three novel genes that had been missed by computational predictions. Moreover, analysis of transcriptional start sites revealed that the average length of the 5' untranslated region was 346 nt, which is much longer than that in humans. The transcriptional start sites of all the genes studied were far more diverse than those of human genes. These observations may reflect unique mechanism(s) of gene expression in this organism, which has an extremely AT-rich genome. PMID- 12095685 TI - Human aldolase C gene expression is regulated by adenosine 3',5'-cyclic monophosphate (cAMP) in PC12 cells. AB - We have examined the effects of an adenosine 3',5'-cyclic monophosphate (cAMP) analog on human aldolase C gene expression in the rat pheochromocytoma cell line PC12. Incubation for 4 h with 500 microM 8-Br-cAMP increased aldolase C mRNA expression 2.5-fold and the expression was still above basal level 24 h later. Using transient transfection experiments we demonstrate that the distal element D in the promoter region of the human aldolase C gene, which binds a transcriptional activator (NGFI-B), is involved in this regulation. NGFI-B mRNA and protein expression were promptly (15 min) increased after 8-Br-cAMP treatment and precedes aldolase C mRNA increase (30 min). After 4 h of 8-Br-cAMP treatment, the binding of NGFI-B protein to the distal element D in the distal promoter region was increased twofold and this correlates with the increased expression of the clone that contains distal element D. These results indicate that the distal element D in the promoter region of the human aldolase C gene is the target of a cAMP-dependent regulation pathway. PMID- 12095686 TI - Genomic structure, organization, and promoter region analysis of murine guanylyl cyclase/atrial natriuretic peptide receptor-A gene. AB - We have determined the complete genomic nucleotide sequence and analyzed the promoter region of murine guanylyl cyclase/natriuretic peptide receptor-A gene (Npr1,coding for NPRA). The gene spans about 17.8 kb and contains 22 exons interrupted by 21 introns. All the exon-intron boundaries possess the consensus GT/AG splice junctions. Four different types of short interspersed nuclear elements (ten mouse B1 elements, seven mouse B2-B4 elements, one ID and one MIR element) and one medium reiteration frequency repeats have been found in the non coding regions of the gene. Eleven tandem repeats, including three in the promoter region of the gene, have been identified. The transcription start site, 362 bp upstream from the start codon, was determined by 5'- rapid amplification of cDNA ends. The 1.98 kb 5'-flanking region contains three potential SP1 binding sites and one inverted CCAAT box but lacks the TATA box. This region also contains several putative cis-acting motifs known to bind kidney specific nuclear protein HFH-3, cAMP-responsive element binding protein (CREB) and AP-4. In addition, the binding sites for a variety of transcription factors: AML-1 alpha, SRY, Nkx-2.5, LyF-1, p300, GATA-1/2, HNF-3 beta, c/EBP alpha/beta and USF have been localized in the promoter region of Npr1 gene. The analyses and characterization of the genomic structure of murine Npr1 gene should yield important insights into the species-specific regulation of this important gene family. PMID- 12095687 TI - Structure, upstream promoter region, and functional domains of a mouse and human Mix paired-like homeobox gene. AB - Mix/Bix proteins represent a vertebrate subgroup of paired-like homeodomain proteins which are known to function around the time of gastrulation. Here we report the structures of the genomic and upstream promoter regions of a mouse and human Mix-like gene. Both genes map to syntenic regions of chromosome 1 and contain two coding exons, with the paired-type homeodomain split between the exons within helix 3. Differentiating mouse embryonic stem cells transcribe a messenger RNA of approximately 2.6 kb. The first exon encodes the translation initiation codon and a 5' untranslated region of approximately 90 bp. Sequence analysis of the 960 bp upstream of the transcription start site of the mouse Mix gene revealed the presence of a putative initiator region and TATA box as well as potential Smad, FoxH1/FAST, T-box, COUP-TF, C/EBP, GATA, HNF3 binding sites and retinoic acid response elements. A number of these sites are conserved in the human Mix promoter. We find that most paired-related homeodomain proteins, including mouse and human Mix, contain a proline-rich region within their amino termini which may interact with other proteins. Mouse and human Mix proteins contain highly conserved carboxy-terminal polar/acidic regions with the potential to form an amphipathic helix and the ability to activate transcription in yeast. Mouse Mix expressed in COS cells or in vitro binds a DNA consensus sequence identified previously for paired class homeodomain proteins. These studies suggest that a number of features of paired-like protein structure and function are conserved across diverse species and provide a useful framework for studying the function and regulation of the mouse Mix gene. PMID- 12095688 TI - The half-life of the transcript encoding the folate receptor alpha in KB cells is reduced by cytosolic proteins expressed in folate-replete and not in folate depleted cells. AB - The KB cell, a transformed human cell line, constitutively expresses a high level of the glycosylphosphatidylinositol (GPI) anchored folate receptor protein alpha (FR alpha) and thereby can grow in medium containing <1 nM folate. When transferred from a folate-replete (FR) medium to one folate-deficient (FD), intracellular folate diminishes about 50-fold and expression of the FR alpha increases 6-fold. This up-regulation is mediated by a 4.5-fold increase in the initial transcription rate and a 2.4-fold prolongation of the mRNA half-life that together provide a higher level of the steady-state mRNA abundance. An RNA gel shift assay of a 5' region of the mRNA that includes all of the non-coding and 24 nt of coding sequence, and a 3' region comprised only of coding sequence, identified unique complexes with cytosolic proteins from the FR-KB cells that were not observed with the cytosol from FD-KB cells. It appears, therefore, that expression of these folate-dependent cytosolic trans-active proteins function to maintain a shorter half-life of the mRNA in the FR-KB cells by binding to 5' and 3' cis elements, reducing the stability of this transcript. PMID- 12095689 TI - Genomic characterization of the RUNX2 gene of Fugu rubripes. AB - A 105 kb Fugu rubripes genomic region containing the RUNX2 ortholog (frunx2) was sequenced and analysed. Spanning 32 kb, frunx2 is seven times smaller than its human orthologue (223 kb). By comparison of Fugu and human genomic environment a stretch of conserved synteny, comprising the neighbouring genes on both sides, was identified. Except one exon that is alternatively spliced in human RUNX2, all other seven exons could be identified in frunx2. The predicted protein sequence of frunx2 shows a high degree of sequence conservation compared with RUNX2 (83% identity). Like all human paralogues, frunx2 possesses two promoter regions separated by a large intron. Both promoter regions are conserved between the two species and contain several RUNX binding sites pointing to a self-regulatory function. Three further conserved non-coding regions were identified possibly functioning as enhancer elements for tissue-specific expression of RUNX2. PMID- 12095691 TI - Translational machinery of channel catfish: I. A transcriptomic approach to the analysis of 32 40S ribosomal protein genes and their expression. AB - Ribosomal protein (RP) genes have become widely used as markers for phylogenetic studies and comparative genomics. However, they have not been available for evolutionary studies in fish although teleosts are the largest group of vertebrates with more than 23,000 species. Using a transcriptomic approach, we have cloned and sequenced 32 40S RP complementary DNAs (cDNAs) from channel catfish (Ictalurus punctatus), making them one of the most complete sets of 40S RP gene sequences from a single organism. Most 40S RPs in channel catfish are highly similar to their orthologues in mammalian species, but S19, S21, and S25 are highly divergent. Only one type of cDNA was found for all RP genes except S26 and S27, for which two cDNAs were found in channel catfish. Alternatively spliced transcripts for the S3 and alternatively polyadenylated transcripts for S19 and S21 were found. The 32 40S RP genes are generally highly expressed and together they account for 5.33-11.42% of expression depending on the tissues. Expression levels of the RP genes were highly variable both within a single tissue among different RP genes and among tissues with regard to a single RP gene. Taken together, these data strongly suggest post-transcriptional regulation of RP gene expression, particularly in consideration of the stoichiometry of their representation in ribosomes. PMID- 12095690 TI - Regulation of transcription of the murine gamma-glutamyl hydrolase gene. Delineation of core promoter A and the role of LYF-1, E2F and ETS-1 in determining tumor-specific expression. AB - Our earlier studies (Gene 268 (2001) 183) showed that transcription of the mouse gamma-glutamyl hydrolase (gamma GH) gene is under the control of two separate promoters widely distributed within the genome. We now report on further studies examining the functional characteristics of the more efficient promoter (promoter A) which is contiguous with an exon 1 (exon A1a) alternate within the main body of the gene. Functional deletion analysis of promoter A in pGL3 transfected in NIH3T3 cells defined a 189 bp region of sequence 26 bp upstream of exon A1a as mediating core promoter transcriptional activity. Further functional deletion analysis and site-directed mutagenesis attributed the activity of this core region to the presence of five SP1 sites within the most upstream 130 bp of sequence and the presence of three cis-active elements, Ets-1, Lyf-1 and E2F in the remaining downstream 59 bp of sequence. Electrophoretic gel-mobility shift assays showed that differences in the binding of trans-acting factors to these three elements accounts for the tumor-specific expression of this gene. Finally, analysis by fluorescent in situ hybridization revealed that promoter A and the main body of the mouse gamma GH gene are located between A3 and A5 on chromosome 4. This is an interesting finding in light of our earlier results which located promoter B and two associated alternates of exon 1 of this gene on chromosome 17. PMID- 12095692 TI - Characterization of a fission yeast subunit of an RNA polymerase I essential transcription initiation factor, SpRrn7h/TAF(I)68, that bridges yeast and mammals: association with SpRrn11h and the core ribosomal RNA gene promoter. AB - Production of eukaryotic ribosomal RNAs (rRNAs) entails sequence-specific recognition of regulatory sequences in the rRNA gene promoter. A putative subunit of the Schizosaccharomyces pombe essential transcription initiation factor for rRNA synthesis has been identified that shares homology with both murine TAF(I)68 and Saccharomyces cerevisiae Rrn7p, subunits of their species' transcription initiation factor. Affinity purified putative SpRrn7h and associated factors, including a putative Rrn11p homolog, SpRrn11h, bear RNA polymerase I transcription initiation factor activity, and recombinant SpRrn7h associates with S. pombe core rDNA promoter sequences. In the first widespread search for putative homologs of SpRrn7h/murine TAF(I)68, and SpRrn11h/murine TAF(I)48, multiple ones were identified across eukaryotes. Analysis of residues conserved between the fission yeast and murine essential initiation factor subunits aided in these identifications. Sequences in the core rRNA gene promoter contributing to transcriptional activation were investigated, including a perfect TATAAA element located at -35. PMID- 12095693 TI - Transcriptional regulation of the mouse fatty acid amide hydrolase gene. AB - Fatty acid amide hydrolase (FAAH) is a membrane-bound enzyme that inactivates a family of fatty acid amide molecules which are implicated in physiological processes such as pain and sleep. We cloned a 1.9 kb fragment of the 5' untranslated region of the mouse FAAH gene into the pGL3 basic luciferase reporter vector and showed that this sequence has promoter activity in vitro. By primer extension analysis, we have determined the transcription start site to be 200 bases upstream of the ATG initiation codon and found that a TATA motif was absent. A number of putative response elements, including those for estrogen and glucocorticoids, were identified in this sequence. We have demonstrated that the estrogen and glucocorticoid receptors down-regulate transcriptional activity independent of their ligand. These data should help in understanding the mechanisms of FAAH gene transcription. PMID- 12095695 TI - Human mitochondrial transcription factor A (mtTFA): gene structure and characterization of related pseudogenes. AB - Mitochondrial transcription factor A (mtTFA or Tfam) is a 25 kDa protein encoded by a nuclear gene and imported to mitochondria, where it functions as a key regulator of mammalian mitochondrial (mt) DNA transcription and replication. The coding sequence of the human mtTFA gene is reported in the literature and the sizes of few introns are known. In this paper we present the genomic structure of the human mtTFA gene along with the complete sequence of its six intronic regions. Three of the introns (I, III, VI) have been found to be less than 600 bp, while the other three were greater than 1.8 kb. In the course of this work, we discovered that, in addition to the active copy, different homologous sequences identified as processed pseudogenes psi h-mtTFA have been isolated and sequenced. Using an 'in silico' mapping approach we determined their locations on chromosomes 7, 11 and X. psi h-mtTFA locations are different from that of the gene, previously reported on chromosome 10. Transcription analysis by means of reverse transcriptase-polymerase chain reaction has shown that other than the RNA corresponding to the full-length transcript, an isoform lacking 96 bp is also present. Among the three sequenced pseudogenes only one of them located on chromosome 11 has been found to be transcribed in Jurkat cells under these culture conditions, even though transcription initiation and binding sites for different transcription factors have also been found upstream from the other two pseudogenes. PMID- 12095694 TI - Differential conservation of transcriptional domains of mammalian Prophet of Pit 1 proteins revealed by structural studies of the bovine gene and comparative functional analysis of the protein. AB - The Prophet of Pit-1 (PROP1) gene encodes a paired class homeodomain transcription factor that is exclusively expressed in the developing mammalian pituitary gland. PROP1 function is essential for anterior pituitary organogenesis, and heritable mutations in the gene are associated with combined pituitary hormone deficiency in human patients and animals. By cloning the bovine PROP1 gene and by comparative analysis, we demonstrate that the homeodomains and carboxyl termini of mammalian PROP1 proteins are highly conserved while the amino termini are diverged. Whereas the carboxyl termini of the human and bovine PROP1 proteins contain potent transcriptional activation domains, the amino termini and homeodomains have repressive activities. The bovine PROP1 gene has four exons and three introns and maps to a region of chromosome seven carrying a quantitative trait locus affecting ovulation rate. Two alleles of the bovine gene were found that encode distinct protein products with different DNA binding and transcriptional activities. These experiments demonstrate that mammalian PROP1 genes encode proteins with complex regulatory capacities and that modest changes in protein sequence can significantly alter the activity of this pituitary developmental transcription factor. PMID- 12095696 TI - The identification of a novel human homologue of the SH3 binding glutamic acid rich (SH3BGR) gene establishes a new family of highly conserved small proteins related to Thioredoxin Superfamily. AB - The SH3 binding glutamic acid-rich (SH3BGR) gene was cloned in an effort to identify genes located to human chromosome 21, within the congenital heart disease region, and expressed in the developing heart. After the identification of SH3BGR, two human homologous genes, SH3BGRL and SH3BGRL3, were identified and mapped to chromosome Xq13.3 and 1p34.3-35, respectively. SH3BGRL and SH3BGRL3 code for small proteins similar to the N-terminal region of the SH3BGR protein. SH3BGRL3 protein shows a significant similarity to Glutaredoxin 1 of Escherichia coli, and all the three proteins are predicted to belong to Thioredoxin-like protein Superfamily. Here we describe the identification and characterization of an additional human homologue of SH3BGR, named SH3BGRL2. The SH3BGRL2 gene maps to chromosome 6q13-15 and its messenger RNA has a large 3' untranslated region containing several AUUUA repeats. SH3BGRL2 codes for a protein of 107 amino acids, which, like SH3BGRL and SH3BGRL3 proteins, is highly homologous to the N terminal region of the SH3BGR protein and appears to be related to Glutaredoxins and to PKC-interacting cousin of thioredoxin homology domain. We propose that the identification of SH3BGRL2 establishes a novel family of human genes, coding for highly conserved small proteins belonging to Thioredoxin-like protein Superfamily. PMID- 12095697 TI - Molecular cloning, mapping and characterization of a novel mouse RING finger gene, Mrf1. AB - With a combined approach of database search, heterologous polymerase chain reaction (PCR), reverse transcription-PCR, rapid amplification of complementary DNA ends and genomic library screening, we have successfully cloned a mouse RING finger gene, mouse RING finger 1 (Mrf1). The Mrf1 gene has two exons of 63 and 2665 bp, respectively, and one intron of over 13 kb. An open reading frame was identified exclusively in exon 2, which encodes a putative protein of the RING-B box-coiled coil or the tripartite motif type of 403 amino acids. Mrf1 is moderately expressed in the spleen, brain and heart as a single 3.0 kb product and very highly expressed in the testis as two transcripts of 3.0 and 1.5 kb, respectively. The Mrf1 gene was mapped to mouse chromosome 3, between markers D3Mit70 and D3Mit277. Western blotting analysis indicated that an expected protein of approximately 44 kD was detected in the brain extracts of mouse, rat and human. The possible functions of Mrf1 are discussed in the contexts of protein-protein interactions, oncogenesis and ubiquitination. PMID- 12095698 TI - Effect of the DNA topoisomerase II inhibitor VP-16 on illegitimate recombination in yeast chromosomes. AB - Etoposide and teniposide, derivatives of podophyllotoxin, are inhibitors of DNA topoisomerase II and are potent anticancer agents. An adverse effect linked to the use of these drugs is the development of acute myeloid leukemia, a disorder usually associated with chromosomal translocation. To examine podophyllotoxin induced DNA rearrangement, we developed an assay system to measure illegitimate recombination in Saccharomyces cerevisiae chromosomes. This approach uses juxtaposed CAN1-CYH2 negative selection markers that are introduced into the LEU2 locus, which is located on chromosome III, in a yeast strain carrying the mutated can1 and cyh2 genes. Upon formation of a deletion over the active CAN1-CYH2 genes, a cell becomes resistant to both canavanine and cycloheximide. To introduce drugs into the cell, we used a yeast strain carrying an ISE2 mutation, thereby making the cell drug-permeable. Here we show that treatment of cells with etoposide (VP-16) increases the rate of illegitimate recombination in yeast, indicating that VP-16 stimulates DNA topoisomerase-mediated illegitimate recombination. Structural analysis of the resulting recombinants indicate that most are formed by deletion mutations on chromosome III, which take place between short homologous regions of DNA. We propose a model for illegitimate recombination, in which VP-16 facilitates formation of a cleavable complex between DNA topoisomerase II and DNA, thus promoting DNA double-strand breakage with the resulting DNA ends joined by a non-homologous mechanism. PMID- 12095699 TI - Distal and proximal cis-linked sequences are needed for the total expression phenotype of the mouse alcohol dehydrogenase 1 (Adh1) gene. AB - Mouse alcohol dehydrogenase 1 (Adh1) gene expression occurs at high levels in liver and adrenal, moderate levels in kidney and intestine, low levels in a number of other tissues, and is undetectable in thymus, spleen and brain by Northern analysis. In transgenic mice, a minigene construct containing 10 kb of upstream and 1.5 kb of downstream flanking sequence directs expression in kidney, adrenal, lung, epididymis, ovary and skin but promotes ectopic expression in thymus and spleen while failing to control expression in liver, eye, intestine and seminal vesicle. Cosmids containing either 7 kb of upstream and 21 kb of downstream or 12 kb of upstream and 23 kb of downstream sequence flanking genetically marked Adh1 additionally promotes seminal vesicle expression suggesting downstream or intragenic sequence controls expression in this tissue. However, expression in liver, adrenal, or intestine is not promoted. The Adh1(a) allele on the cosmid expresses an enzyme electrophoretically distinct from that of the endogenous Adh1(b) allele, and presence of the heterodimeric enzyme in expressing tissues confirms that transgene activity occurs in the same cell-type as the endogenous gene. Transgene expression levels promoted by cosmids were at physiologically relevant amounts and exhibited greater copy-number dependence than observed with minigenes. Transgene mRNA expression correlated with expression measured at the enzyme level. A bacterial artificial chromosome containing 110 kb of 5'- and 104 kb of 3'-flanking sequence surrounding the Adh1 gene promoted expression in tissues at levels comparable to the endogenous gene most importantly including liver, adrenal and intestinal tissue where high level Adh1 expression occurs. Transgene expression in liver was in the same cell types as promoted by the endogenous gene. Although proximal elements extending 12 kb upstream and 23 kb downstream of the Adh1 gene promote expression at physiologically relevant levels in most tissues, more distal elements are additionally required to promote normal expression levels in liver, adrenal and intestinal tissue where Adh1 is most highly expressed. PMID- 12095700 TI - Major intestinal coactivator p300 strongly activates peroxisome proliferator activated receptor in intestinal cell line, Caco-2. AB - We have previously reported that several genes related to intestinal fatty acid and vitamin A metabolism are coordinately regulated by peroxisome proliferator activated receptor (PPAR) [Arch. Biochem. Biophys. 389 (2001) 41; Biochim. Biophys. Acta 1531 (2001) 68]. In this study, we demonstrated that PPAR alpha and PPAR delta interacted with endogenous coactivators in intestinal cell line, Caco 2 in a ligand specific manner. We isolated rat cDNA clones encoding the nuclear receptor interaction domains of the two transcriptional coactivators, CREB binding protein (CBP) and p300. Expression level of CBP mRNA was relatively low in the small intestine, while p300 mRNA was ubiquitously expressed in various tissues including the small intestine in the rat. Southern blot analysis revealed that these coactivators were encoded by different genes. Mammalian two-hybrid assays in Caco-2 cells revealed that p300 interacted with PPAR alpha or PPAR delta in the presence of their specific ligands more efficiently than CBP did. These results suggest that the major intestinal coactivator, p300 strongly interacts with PPAR alpha and PPAR delta. PMID- 12095701 TI - cDNA cloning, expression pattern and RNA binding analysis of human selenocysteine insertion sequence (SECIS) binding protein 2. AB - Selenocysteine and selenoprotein synthesis require a complex molecular machinery in mammals. Among the key players is the RNA-protein complex formed by the selenocysteine insertion sequence (SECIS) binding protein (SBP2) and the SECIS element, an RNA hairpin in the 3' untranslated regions of selenoprotein messenger RNAs (mRNAs). We have isolated the DNA complementary to mRNA of the human SBP2, enabling us to establish that it differs from a previously reported human SBP2 like protein. Examination of the expression pattern revealed that the human SBP2 protein is encoded by a 4 kb long mRNA that is over-expressed in testis. Compared to the rat SBP2 sequence, the human SBP2 protein displays two highly conserved domains with 92 and 95% amino acid identity, the latter one containing the RNA binding domain. The inter-domain section carries 55% sequence identity, the remainder of the SBP2 sequences showing about 65% identity, values lower than expected for two mammalian proteins. Interestingly, we could show that the binding of human SBP2 to the SECIS RNA is stimulated by the selenoprotein specialized elongation translation factor mSelB/eEFsec. PMID- 12095703 TI - Identification of a novel HLA-B-35-restricted cytotoxic T-lymphocyte epitope within human immunodeficiency type 1 (HIV-1) P17(Gag). PMID- 12095704 TI - Immunological consequence of sulfur mustard exposure. AB - Global reports indicate that chemical weapons still impose a serious threat to world security and health. Sulfur mustard is a chemical compound with devastating short and long-term effect on human health. PMID- 12095702 TI - Structural characterization of Rasgrf1 and a novel linked imprinted locus. AB - Imprinted genes in mammals are expressed either from the maternally or the paternally inherited allele. Previously, a genome wide scan identified novel imprinted genes based on their association with differentially methylated regions (DMRs). One of the identified genes, Rasgrf1, showed paternal expression in neonatal brain and was located on mouse chromosome 9. This gene is associated with a DMR, located about 30 kb upstream of Rasgrf1 exon 1. In order to better understand and identify novel elements involved in the regulation of this gene we have isolated and characterized genomic clones coding for mouse and human Rasgrf1 and RASGRF1, respectively. The mouse gene consists of 26 exons spanning approximately 140 kb of genomic DNA while the human gene has 28 exons. The human gene has an additional 39 bp exon inserted between exons 13 and 14 and exon 18 is split in two separate exons in human. The major transcription start site of Rasgrf1, as identified by primer extension, is 1324 bp upstream of the ATG translation start codon. Finally, a genomic region upstream of exon 1, spanning 489 bp, was determined to possess the essential promoter activity for Rasgrf1 gene. A second gene, A19, located 10 kb upstream of the DMR has been characterized. A19 is mainly expressed in testis and at lower levels in neonatal and adult brain tissue. The A19 transcript is non-coding and expressed in mouse testis and brain. A19 is imprinted with expression occurring from just the paternal allele in brain. PMID- 12095705 TI - Peptide immunomodulators versus infection; an analysis. AB - A disease gets manifested only when the host immune system is not strong enough to fight off the infective agents. A number of small peptides both from natural and synthetic origin are found to be capable of modulating the immune response. While immune adjuvants are known to strengthen the immune response and help the host not to give way to the pathogens thereby preventing their establishment, the immunosuppressors are found useful in autoimmune conditions as well as in facilitating the organ transplants. Recent understanding of immune network, however, reveals its cross connectivity with the endocrine and central nervous systems as well. Thus, the inhibition and control of disease by planned restoration of homeostatis in these systems through immunomodulation is also possible. PMID- 12095706 TI - Regulation on maturation and function of dendritic cells by Ganoderma lucidum polysaccharides. AB - Ganoderma lucidum polysaccharides (Gl-PS) exhibits a variety of immunomodulatory activities, and dendritic cells (DC) are professional antigen presenting cells, which are pivotal for initiation of primary immune response. In this study, the regulatory effects of Gl-PS on maturation and function of cultured murine bone marrow derived DC were investigated in vitro. Gl-PS (0.8, 3.2, or 12.8 microg/ml) could increase the co-expression of CD11c and I-A/I-E molecules on DC surface, promote mRNA expression of cytokine IL-12 p40 in DC and augment protein production of IL-12 P40 in culture supernatants. The lymphocyte proliferation of mixed lymphocyte culture (MLC) induced by mature DC was enhanced by Gl-PS, either. Gl-PS was shown to promote not only the maturation of cultured murine bone marrow derived DC in vitro, but also the immune response initiation induced by DC. PMID- 12095707 TI - Differentiation of the human monocyte cell line, U937, with dibutyryl cyclicAMP induces the expression of the inhibitory Fc receptor, FcgammaRIIb. AB - FC receptor for IgG receptor (Fcgamma) mediated activation of macrophages is essential for the clearance of immune complexes and control of inflammation. However, activated macrophages play an integral role in tissue destruction associated with autoimmune and inflammatory disease processes. Understanding the mechanisms which balance activating and inhibitory signals generated by immune complexes are therefore of critical importance to human disease. Here, we demonstrate that differentiation of the human monocytic U937 cell line to a macrophage phenotype with dibutyryl cyclicAMP induces both mRNA and protein expression of the inhibitory IgG receptor, FcgammaRIIb1. We further demonstrate that, following receptor aggregation, FcgammaRII transiently recruits the 5' inositol phosphatase, SHIP. These data define a role for FcgammaRIIb in the modulation of immune complex mediated macrophage activation in a human model system. PMID- 12095708 TI - Inhibition of arachidonic acid release from human peripheral mononuclear cells by heat shock treatment and geldanamycin. AB - The objective of this study was to investigate the effects of heat shock (HS) treatment and geldanamycin (GA) on the release of arachidonic acid (AA) from human peripheral blood mononuclear cells (PBMC), monocytes and lymphocytes. Mononuclear cells prepared from blood of healthy subjects were preincubated with (3)H-AA. The release of (3)H-AA incorporated into the membrane was studied after pretreatment of cells by HS (43 degrees C, 1 h) and GA. The activation of AA producing enzymes was achieved by the addition of phorbol 12-myristate 13-acetate (PMA) or by the combination of PMA+calcium ionophore A-23187. Treatment of cells by HS inhibited the release of AA. Furthermore, the release of AA by PBMC was dose dependently inhibited by GA. The combination of treatments by HS and GA augmented the inhibition of AA release. The HS response involves a diminished release of AA from PBMC. The inhibitory effect of GA on the AA release is a new element in the antiinflammatory pharmacological ability of this drug. PMID- 12095709 TI - Characterization of monoclonal antibodies and epitope mapping of the NS4 protein of hepatitis C virus. AB - Recombinant DNA containing sequences of HCV NS4 protein was expressed in Escherichia coli cells. Six hybridoma clones producing monoclonal antibodies (MAB) to recombinant NS4 protein (rNS4), aa 1677-1756, were developed. Mapping with a panel of 33 peptides and reciprocal competitive EIA have shown that MAB obtained revealed five antigen determinants, not described earlier: MAB 3F11 and 3F12-one genotype-independent epitope of NS4A (aa 1700-1707) common for genotypes 1, 2 and 3; MAB 1D11-genotype-independent epitope (aa 1713-1728) and MAB 1D3 genotype (subtype 1b)-specific epitope of NS4B (aa 1711-1731); MAB 6B11 and C1 two conformation-dependent determinants in 5-1-1 region. These data indicate that the 5-1-1 region of NS4 protein has a complex antigenic structure and contains at least eight epitopes, including five, revealed in the present work. MAB obtained recognized native viral protein in the cytoplasm of liver cells of patients with chronic hepatitis C. The positive rates of the immunostaining for NS4 antigen using MAB 6B11, 1D11 and 3F12 were 64, 59 and 50%, respectively. It was found that 6B11 MAB to a conformation-dependent epitope much more actively interacts with native NS4 than with the recombinant protein to which MAB was developed. The epitope recognized by 6B11 MAB is highly immunogenic since it induces the B-cell response in all patients investigated with identified anti-NS4 antibodies in blood serum. The MAB panel obtained in this study may become a useful tool for the diagnostic purposes, for the investigation of NS4B function and for the host viral interactions at the cell level. PMID- 12095710 TI - Regulation of the expression of mouse TAP-associated glycoprotein (tapasin) by cytokines. AB - The expression of antigen presenting MHC class I molecules can be enhanced through cytokines, e.g. upon infection with bacteria or viruses, either directly by enhancing class I gene transcription or by increasing the amounts of accessory proteins of the loading complex. Tapasin plays a significant role in the peptide loading of class I molecules. Here, we describe recognition motifs of cytokine inducible transcription factors in the promoter region of the mouse tapasin gene, most of them clustered within the 140 base pairs upstream of the start codon. Tapasin mRNA was strongly induced in vivo after infection with the facultatively intracellular bacterium Listeria monocytogenes in an IFN-gamma-dependent fashion. Accordingly, both tapasin mRNA and protein were strongly induced in a time and dose dependent manner in embryonic fibroblasts treated with the cytokines IFN gamma and IFN-beta, and weakly induced after treatment with TNF-alpha. Co stimulation of tapasin by TNF-alpha and IFN-gamma resulted in a weak synergistic effect. Using fibroblasts either lacking IRF-1 or inhibited in protein synthesis we show that secondary transcription factors are necessary for a maximal stimulation of tapasin expression upon IFN-gamma stimulation. The sequential induction of TAP1, LMP2, and tapasin before the stimulated expression of class I heavy chain is discussed. PMID- 12095712 TI - Microanatomical localization of PD-1 in human tonsils. AB - PD-1 is an immunoinhibitory receptor, which belongs structurally to the CD28 family. PD-1-deficient mice show breakdown of peripheral tolerance and manifest multiple autoimmune symptoms. We previously described expression of PD-1 on activated T and B lymphocytes and myeloid cells. However, little is known about the microanatomical distribution of PD-1 in lymphoid organs. In this study, we performed immunohistochemistry using monoclonal antibodies against human PD-1. In human tonsils, PD-1 was expressed on most of T cells and a small subset of centrocytes in the light zone of germinal centers (GCs), where clonal selection of centrocytes takes place. These results suggest that PD-1 may play an important role in GC reaction. PMID- 12095711 TI - Production of interleukins 10 and 12 by activated peripheral blood monocytes/macrophages in patients suffering from chronic hepatitis C virus infection with respect to the response to interferon and ribavirin treatment. AB - Circulating monocytes/macrophages are important for the initiation of immune responses to hepatitis C virus (HCV). Their presentation capacities and production of immunoregulatory cytokines enable them to activate cellular immune responses which is critical in determining the outcome of infection. We used flow cytometry to examine the expression of a CD80 costimulatory molecule on the surface of peripheral blood CD14+ monocytes/macrophages and to analyse the production of IL10 and IL12 by these cells. Forty-three individuals (6 asymptomatic HCV carriers, 37 patients with chronic hepatitis C (CHC)) were enrolled in this study. Thirty-seven patients with CHC (23 responders and 14 non responders, NR) received combination (interferon+ribavirin) treatment for 52 weeks. The baseline percentage of CD14+CD80+ peripheral blood monocytes/macrophages was high in patients with CHC (P<0.001) and returned to normal after the treatment. All patients with CHC showed significantly high production of IL10 (P<0.001). In asymptomatic HCV carriers production level of this cytokine tended to be higher than in patients with CHC (P<0.001). A baseline production of IL12 was higher in asymptomatic HCV carriers and patients with CHC compared to healthy controls (P<0.001). The level of IL12 production was increased in treatment responders whereas in NR returned to normal value. Our data argue against functional impairment of circulating monocytes/macrophages during HCV infection. Furthermore, the positive therapeutic outcome following combination treatment might associate with increased production of IL12 by these cells. PMID- 12095713 TI - Cellular specificity related to monoglyceride-induced cell death. AB - We have recently observed that monoglycerides (MGs), a family of lipids consisting of a single fatty acid moiety attached to a glycerol backbone, induce rapid dose-dependent apoptosis in murine thymocytes. In this work, we evaluated the sensitivity of various normal and malignant immune and non-immune cells to MGs. We demonstrate that the propensity to MG-induced death displayed by both T and B lymphocytes is clearly modulated according to their differentiation and activation status. For instance, the earliest T and B cell precursors are refractory to MG-mediated cell death. In the T-cell lineage, immature thymocytes are the most susceptible to MG treatment, while B cells from peripheral lymphoid organs appear more sensitive than B-cell subsets from the bone marrow. On the other hand, both activated T and B cells are more resistant to MG exposure than their non-activated counterparts. In addition, other hematopoietic lineages such as natural killer cells, macrophages, and erythroid cells are quite resistant to MG-induced death. Furthermore, using various immortalized cell lines from different tissues, we found that lymphomas and thymomas are the most sensitive among all lineages tested, while epithelial cells and fibroblasts are unaffected by MG treatment. Finally, MG-induced death was shown to be independent of Fas/Fas ligand (FasL) interactions. Altogether, our findings indicate that there is a cellular specificity related to MG-mediated cell death biased towards T and B lymphocytes. This suggests that MGs could potentially be used in the treatment of specific lymphoid disorders by bypassing the requirement for the Fas/FasL system. PMID- 12095714 TI - Sequence variation and consensus sequence of V3 loop on HIV-1 gp120. AB - Mutation in the V3 loop of HIV-1 gp120 could affect syncytium formation, virus infectivity and neutralization. To acquire more information of the V3 loop mutation, we analyzed amino acid sequences of the V3 loop of 24504 isolates from most HIV-1 clades (including A, B, C, D, E, F, G and H clades). The consensus sequence of the V3 loop of each subtype with the highest frequency emerging on each position is constituted and the conservation of each amino acid in this region is also calculated. Exploring the restricted mutation of the V3 region could help to understand mechanism of HIV entry and to develop new strategy against HIV-1. PMID- 12095716 TI - Outcome of hip fracture in older Irish women: a 2-year follow-up of subjects in a case-control study. AB - To assess outcome after hip fracture in older Irish women, 106 consecutive females aged over 50 years admitted to a general hospital with a hip fracture were compared to 89 age- and gender-matched controls from the same catchment area. Interview-based data were collected on socio-demographic factors, mobility and activities of daily living before recruitment and 2 years later. Information was also collected on residence, further falls and fractures and use of health and community support services during the 2-year period. Mortality at 2 years was higher in cases (23.6%) compared to controls (10.1%; P = 0.01). Cases were significantly less mobile and more dependent in the activities of daily living. Of the cases who were community dwellers at baseline, 26.6% were institutionalised at 2 years compared with 9.2% of controls (P = 0.01). During the 2 years cases were significantly more likely to have multiple falls and a further hip or pelvic fracture. Hospital and nursing home admissions and use of physiotherapy, day centre and home help services were also significantly greater among cases. The marked adverse impact of hip fracture reported in this study underlines the importance of public health strategies to prevent these injuries in older people. PMID- 12095715 TI - A survey of the treatment of displaced intracapsular femoral neck fractures in the UK. AB - A survey was undertaken to investigate the treatment of displaced intracapsular femoral neck fractures across the UK. The usual practice at 223 hospitals was recorded for two groups of patients, active and frail. Management of stereotyped fractures, in similar patients, varied between hospitals. There was also variation between specialists within some hospitals: two or more of the alternative methods of treatment were in routine use for active patients at 22% of hospitals and for frail patients at 27%. Overall, for active patients, bipolar hemiarthroplasty was undertaken at 41% of hospitals, internal fixation at 37%, unipolar hemiarthroplasty at 32% and total hip replacement at 16%. Cemented prostheses were used in 74% of arthroplasties for active patients. For frail patients, hemiarthroplasty with an Austin Moore or Thompson prostheses was undertaken at 94% of hospitals; bipolar prostheses were used at 8%; internal fixation was undertaken at 1%. Cement was used in 46% of hemiarthroplasties. These findings indicate a lack of consensus in aspects of the treatment of displaced intracapsular femoral neck fractures and are likely to reflect difficulties in determining "best practice." PMID- 12095717 TI - Sequential hip fractures in elderly patients. AB - Osteoporotic hip fractures are an increasing cause of disability to elderly patients and expenditure to the NHS. The aim of this work was to determine whether the rate of sequential hip fractures in elderly patients has changed over the past 20 years. The design was to do a retrospective study of 186 patients admitted with a diagnosis of femoral neck fracture in order to determine how many had suffered a previous contralateral hip fracture. The results indicated that the number of patients who had had previous surgery for a contralateral hip fracture was 11.8%. In conclusion, the rate of sequential hip fractures in elderly patients has not changed significantly over the past two decades. PMID- 12095718 TI - The AO/ASIF proximal femoral nail (PFN) for the treatment of unstable trochanteric femoral fracture. AB - The AO/ASIF proximal femoral nail (PFN) is a new device designed for the treatment of the unstable trochanteric femoral fracture. This study reports the outcome in such fractures treated using the PFN at a District General Hospital. All patients presenting to our department with unstable trochanteric femoral fractures were treated operatively using the PFN. A total of 76 patients were included in the study and were followed up to fracture union or fixation failure. A case documentation form and follow-up form were used to collect the data which included the Salvati and Wilson assessment of hip function. The majority of the procedures were reported by the operating surgeon as "easy" or "usual". Distal locking was difficult in three patients. In one patient, the fixation failed because the screws were wrongly positioned and was revised to a THR. Mortality rate, during the first 3 months, was 27%. Of the surviving patients, screws cut through the femoral head in four patients (8%), however, fractures united in all the patients. There was one incidence of fracture around the tip of the nail. Seventy-eight percent of the patients at the final follow-up scored >20 points (out of 40 points), using the Salvati and Wilson hip function scoring system. According to the patients and/or their carers, outcome was described as good or very good in 94% of the patients and the level of function was similar to pre injury level in 50% of the patients. We conclude that the PFN is a useful device in the treatment of the unstable trochanteric femoral fracture. It is a relatively easy procedure and a biomechanically stable construct allowing early weight bearing. Femoral neck screws positioning is critical. PMID- 12095719 TI - The treatment of unstable, extracapsular hip fractures with the AO/ASIF proximal femoral nail (PFN)--our first 60 cases. AB - The most widely used implant in unstable proximal femoral fractures is the dynamic hip screw (DHS). Intramedullary implants include the intramedullary hip screw (IHMS), gamma nail and proximal femoral nail (PFN). There have not yet been any randomised trials comparing the PFN and DHS. We report our experience of stabilising 60 consecutive proximal femoral fractures with the PFN. Fifty fractures were unstable trochanteric fractures, seven were subtrochanteric (32A), one segmental, and two were at the level of a DHS plate. The mean age was 79 years and there were 12 male and 48 female patients. Twenty-eight patients were graded as ASA 3 or 4. Two patients had multiple injuries, all fractures were closed and there were five pathological fractures. The patients were followed up for a minimum of 4 months. Within this period, there were 12 deaths due to cardiopulmonary complications. Of the remaining 48 patients, two were lost to follow-up and 39 had united by 4 months. The local complications were: four implant cut-outs, two high energy fractures below the implants and one implant failure at 7 months due to delayed union. We believe that the PFN is a good choice for stabilising subtrochanteric fractures. We also believe that the use of the PFN for unstable trochanteric fractures is very encouraging. A large, randomised controlled trial against the DHS would be helpful to clarify the relative risks and benefits. PMID- 12095720 TI - IMHS clinical experience in the treatment of peritrochanteric fractures. The results of a multicentric Italian study of 981 cases. AB - This retrospective study evaluates the results obtained in five Italian departments of traumatology in the treatment of peritrochanteric (pertrochanteric and subtrochanteric) fractures by the intramedullary hip screw (IMHS; Smith & Nephew Richards, Memphis, TN, USA) nail. One thousand two hundred and seventy three patients were treated with the IMHS nail between March 1992 and February 2000. The results of these operations were evaluated clinically and radiological in 981 patients. The 90.3% of patients could walk unaided or with simple support. Because of the low complication rate requiring re-operation (postoperative shaft fractures, screw penetrated the acetabulum, cut out and non-union) (1.7%), we think that this device is an advance in the treatment of peritrochanteric fractures. PMID- 12095722 TI - Austin Moore hemiarthroplasty: technical aspects and their effects on outcome, in patients with fractures of the neck of femur. AB - In order to determine which technical aspects of the Austin Moore hemiarthroplasty procedure affect the outcome, we reviewed 243 patients with a non-pathological intracapsular femoral neck fracture who had, Austin Moore uncemented hemiarthroplasty. The immediate post-operative X-rays were assessed for alignment of the prosthetic stem, calcar seating, length of the neck remnant, leg length discrepancy and size of the head, compared with the contralateral femur. All patients were followed-up for 1 year. Significant pain at 1 year and/or revision of the prosthesis for loosening were considered as unfavourable outcomes. Inadequate calcar seating was significantly associated with pain and revision of the prosthesis (P = 0.04 and 0.01, respectively). Length of the neck remnant was also significantly associated with these two outcomes (P = 0.05 and 0.023, respectively). Difference in head size was associated with pain, but not with loosening (P = 0.01 and 0.08, respectively). The rest of the parameters were not significantly associated with the outcome. We recommend that when inserting an Austin Moore hemiarthroplasty, particular attention must be paid to the seating of the collar of the prosthesis on the calcar and correct choice of head size. PMID- 12095721 TI - Percutaneous compression plating (PCCP) versus the dynamic hip screw for pertrochanteric hip fractures: preliminary results. AB - The percutaneous compression plate (PCCP) is a new implant for the minimally invasive treatment of pertrochanteric hip fractures that might reduce blood loss, wound problems and prevent devascularization of bone fragments. A quicker operation with minimal blood loss is better in the older patients. We performed a prospective, randomized clinical trial to compare the PCCP with the well-known dynamic hip screw (DHS). A total of 71 patients with an Evans type 1A-D pertrochanteric hip fractures were included. We measured the operation duration, blood loss, wound healing, complications, fracture healing and functional outcome. In total, 33 PCCP and 38 DHS were implanted. The mean operation times were 69.2 and 46.6 min for DHS and PCCP, respectively (P = 0.000). Blood transfusions were given in 24 DHS patients compared with six PCCP patients (P = 0.000). There were 27 haematomas in the DHS group and eight in the PCCP group (P = 0.000). There were no differences in fracture healing and the functional outcome between the two implants (P = 0.767, ns). Although this is a preliminary study with a relatively small number of patients and short follow-up, the PCCP seems similar to the DHS in relation to bone healing and stability, but with significant advantages for blood loss, soft tissue healing and operation time. PMID- 12095723 TI - Austin-Moore hemiarthroplasty for failed osteosynthesis of intracapsular proximal femoral fractures. AB - We studied the outcome of 100 uncemented Austin-Moore hemiarthroplasties used as a revision procedure for failed osteosynthesis of intracapsular femoral fractures. This group was compared with 730 patients in whom an uncemented Austin Moore prosthesis had been used as the primary treatment for an intracapsular femoral fracture. The results indicate that the study group had more pain at 1 year post-fracture. There were also significantly more revision procedures in those who had the arthroplasty performed as a salvage procedure. In general, the results of uncemented Austin-Moore hemiarthroplasty used as a revision procedure for failed osteosynthesis are inferior to that for primary hemiarthroplasty. We would advocate that for any case in which there are signs of acetabular damage a total hip replacement might be a better procedure. For the remainder, an alternative arthroplasty should be considered with an uncemented Austin-Moore prosthesis reserved for only the very frail. PMID- 12095724 TI - Predictive geometric factors in a standardized model of femoral neck fracture. Experimental study of cadaveric human femurs. AB - An experimental study was performed on 20 cadaveric human proximal femurs to investigate both the reproducibility of their mechanical behavior under uniaxial compression and the correlation of mechanical properties with geometric dimensions and bone density. These variables were assessed by radiography, physical measurement, and dual-energy X-ray absorptiometry (DEXA). The specimens were immobilized, loaded to the point of fracture, and analyzed with the help of a materials testing machine. The fractures invariably showed a similar pattern both in location (i.e. at the femoral neck) and in nature (i.e. "axial-shear" type according to the AO classification system). The mechanical properties of the tested proximal femurs correlated negatively with age (r = -0.39) and positively with sex (male femurs were stronger and correlated more closely than female femurs, P = 0.005). They showed a strong positive correlation with head diameter (r = 0.713, P = 0.0004), a moderate positive correlation with both neck axis length (r = 0.63) and neck diameter (r = 0.502), and a slight positive correlation with femoral neck-shaft angle (r = 0.326). All DEXA-based densitometry measures turned out to be powerful predictors of fracture force. The establishment of this solidly reproducible fracture model based on standardized loading conditions should have implications for future research on osteopenia/osteoporosis and preventive stabilization techniques. PMID- 12095725 TI - The role of laser Doppler flowmetry in assessing the viability of bone fragments in an open fracture. AB - The management of open fractures requires excision of all devitalised tissues, both bony and soft tissue, and failure to do so is likely to increase the risk of infection. This study evaluated the applicability of laser Doppler flowmetry for the objective evaluation of fracture fragment viability in an experimental open ballistic fracture over a period of 12 h. The results indicate that this technique could not be used to distinguish between vascularised and non vascularised fragments at any time, and did not aid the surgeon in their decision making at the time of wound excision. Subjective evaluation, based upon the degree of soft tissue attachment of fragments, was a far better indicator of fragment vascularity, although it had a relatively low specificity. There remains the need for education and training for trauma surgeons in the evaluation of fragment viability to ensure adequate wound excision as part of fracture management. PMID- 12095726 TI - Pathomechanism of shock wave injuries on femoral artery, vein and nerve. An experimental study in dogs. AB - PURPOSE: To study the pathomechanism of shock wave injuries to the femoral artery, vein and nerve in dogs. MATERIALS AND METHODS: High-energy shock wave (0.47 mJ/mm(2) energy flux density) were applied to the right femoral artery, vein and nerve in nine mongrels, and the left side was used as control. Macroscopic examinations including structure integrity, edema, discolouration and extravasation; and microscopic examinations including detachment of the intima layer, disruption of the medium layer, capillary congestion, neutrophil margination and extravasation of the outer layer and the surrounding tissues were performed in 2 h, and in 4 days, respectively, after shock wave application. RESULTS: The most serious injuries included disruption of the medium layer with separation from the adventitia layer of the femoral artery. The injuries to the femoral vein and nerve predominantly involved the outer adventitia layer and the surrounding tissues. Mild nerve bundle swelling was noted in most cases. There were capillary congestion, neutrophil margination and extravasation indicative of inflammatory tissue reaction after shock wave application. CONCLUSION: High energy shock waves caused serious injuries to the femoral artery, vein and nerve, especially the femoral artery and inflammatory reaction to the surrounding tissues. PMID- 12095727 TI - Bone blood flow response to surgical trauma. AB - The evolution of bone plate design has been with a view to reducing the interface contact between the plate and the underlying bone thereby limiting the perfusion deficiency that develops. Little consequence, however, has been attributed to the drilling of holes in the steps prior to bone plate application; the work present herein attempts to define the vascular response of bone to the trauma of drilling holes. Anaesthetized sheep underwent the creation of drill holes in both tibiae and metatarsi. Animals were then heparinized and euthanatized. Utilizing femoral cannula, perfusion of the vasculature ensued; spalteholz (India ink), disulphine blue and radiocontrast material (barium sulfate). Decalcified histology was performed and correlated with the perfusion studies. Regions of perfusion deficiency were observed immediately adjacent to, and removed from, the drill hole site. Radiographic images showed substantial haematoma formation and vascular disruption in the vicinity of the drill hole site. Histologically, blood vessels immediately adjacent and distal to the drill hole site, contained a proteinaceous/cellular material occluding the vascular space. Our findings support the proposal of a short-term obstruction in cortical blood flow which may contribute to later adaptational osteopaenia following bone plate application. We have observed an acute vascular insufficiency in cortical bone directly related to the trauma of drilling screw holes. PMID- 12095728 TI - Gunshots to the neck: selective angiography as part of conservative management. AB - Trauma units all over the world are faced with an ever-increasing number of gunshot injuries. While the traditional view is that exploration is mandatory for all gunshot wounds to the neck, this issue is now often debated amongst trauma surgeons. The aim of this particular study was to assess the outcome of gunshot wounds to the neck using a selective conservative approach. Haemodynamically stable patients were investigated with angiography. Only when this proved to be positive, the patient was surgically explored. The records of 116 patients presenting with a gunshot to the neck to our trauma unit over a 3-year-period were reviewed. We studied demographics, bullet track, clinical findings, diagnostic investigations, methods of treatment, time in hospital and outcome. Seventy of the 116 patients sustained a direct hit to the neck, in 46 patients the bullet traversed the face or chest first. Eighty-five patients presented with vascular injury, 61 with an injury to the airway, 32 with an injury to the pharynx or oesophagus, and 12 with sustained neurological damage. Angiography was performed in 89 patients and was positive in 12 patients. Lesions occurred in the common carotid artery (seven), the internal carotid artery (three), the external carotid artery (three), the vertebral artery (two) and the subclavian artery (one). Five patients had more than one lesion. In total 18 patients were treated operatively by performing a neck exploration. Four patients had emergency surgery for exsanguinating bleed. Fourteen had surgery after a positive diagnostic study; 12 after angiography, 2 after another positive investigation. Ten (8.6%) patients died; three during resuscitation, three during emergency exploration, two due to respiratory failure, one postoperative and one from the adult respiratory distress syndrome (ARDS). Our results suggest that selective conservative management is a good treatment for gunshot wounds of the neck. In our experience angiography plays a key role in the detection of a major vascular injury requiring surgical exploration. Careful clinical assessment enhanced with the appropriate investigations is the cornerstone for deciding to explore a gunshot wound to the neck. PMID- 12095729 TI - Unrecognised septic arthritis following fracture of the proximal femur in patients awaiting surgery. PMID- 12095730 TI - Minimally invasive plate osteosynthesis (MIPO) combined with distraction osteogenesis in the treatment of bone defects. A new technique of bone transport: a report of two cases. AB - This study reports two cases of fracture of the femur with segmental bone loss that were treated by a new technique of bone transport. A minimally invasive plate osteosynthesis (MIPO) bridge from the proximal to distal femur served as a fixation system. A separate Wagner distraction apparatus was used for distraction osteogenesis. After the transport segment docked with the bone fragment, it was fixed to the plate by a percutaneous screw fixation. The Wagner lengthening device was then removed. The total time of external fixation was significantly reduced. A satisfactorily functional result and bone segment regeneration was achieved. The external fixation index was 0.31 and 0.37 month/cm, and the limb length discrepancy was 0.9 and 1.2cm. The advantages of this technique include a decrease in the duration of external fixation, protection against refracture and earlier rehabilitation. PMID- 12095801 TI - Wavefront analysis in eyes with nuclear or cortical cataract. AB - PURPOSE: To compare the higher-order aberrations of the oculus (whole eye) and cornea in eyes with mild cortical or nuclear cataract and to estimate the effect of ocular higher-order aberrations on the loss of contrast sensitivity using wavefront analysis. DESIGN: Observational case series. METHODS: Six eyes of four patients with mild nuclear cataract, 18 eyes of 14 patients with mild cortical cataract, and nine eyes of nine normal patients were examined. Wavefront aberrations of the oculus and cornea for central 6 mm diameter were measured using the Hartmann-Shack (HS) aberrometer. Higher-order aberrations were calculated with Zernike polynomials up to sixth order. The relationship between average lens density (ALD) measured by the Scheimpflug camera and the ocular total higher-order aberration (OTHA) was investigated. The relationship between contrast sensitivity (CS) and the OTHA or ALD was also examined. RESULTS: The OTHA was significantly larger in cataracts compared with normal subjects, while corneal total higher-order aberration did not differ between cataracts and normal subjects. The polarity of spherical aberration was negative in all eyes with nuclear cataract while positive in all eyes with cortical cataract. The correlation between ALD and OTHA was not significant in eyes with cataracts. The CS highly correlated with OTHA while it moderately correlated with ALD. CONCLUSIONS: The HS aberrometer is useful to objectively evaluate the deterioration of images in eyes with mild cataract and it revealed that the polarity of spherical aberration was different between nuclear and cortical cataract. It was also suggested that in mild nuclear or cortical cataract, not only light scattering, but also optical aberration of the lens contributes to the loss of contrast sensitivity. PMID- 12095802 TI - Posterior capsule opacification after cataract surgery in patients with diabetes mellitus. AB - PURPOSE: To examine the degree of posterior capsule opacification (PCO) after cataract surgery in patients with diabetes and in nondiabetic age-matched control patients. DESIGN: Case-control study. METHODS: The PCO density value in 100 consecutive diabetic patients and in 100 nondiabetic age-matched control patients who underwent cataract surgery was measured using a Scheimpflug videophotography system at 1 week and at 3, 6, 12, 18, 24, 30, and 36 months after surgery. The incidence of neodymium:yttrium-aluminum-garnet (Nd:YAG) laser posterior capsulotomy was also evaluated. Furthermore, the stage of diabetic retinopathy and systemic condition of the diabetes were correlated with the degree of PCO. RESULTS: No significant difference between the two groups was observed in the mean value of the PCO for up to 12 months after cataract surgery. However, at 18 months and later, the PCO value in the diabetic group increased significantly and was significantly greater than in the control group. Kaplan-Meier survival curves showed that diabetic patients were significantly more likely to require Nd:YAG capsulotomy than control patients (P =.0139, Mantel-Cox log rank test). Among the diabetic patients, there was no significant correlation of PCO value with the stage of retinopathy. Furthermore, type of treatment, duration of diabetes, and hemoglobin A(1C) did not correlate with the PCO value. CONCLUSIONS: Diabetic patients developed significantly greater PCO after cataract surgery than did nondiabetic patients, but among the diabetics, the stage of diabetic retinopathy and systemic status of the diabetes did not correlate with the degree of PCO. PMID- 12095803 TI - Pathological study of cases with secondary open-angle glaucoma due to sarcoidosis. AB - PURPOSE: To investigate the cause of secondary open-angle glaucoma due to sarcoidosis. DESIGN: Observational case series studied by histological methods. METHODS: Seven trabeculectomy specimens from six patients with secondary open angle glaucoma due to ocular sarcoidosis (trabeculectomy group) and anterior parts of seven autopsy eyes from four patients (autopsy eye group) diagnosed as confirmed sarcoidosis were processed for light and transmission electron microscopy. Pathological changes of outflow routes were investigated. RESULTS: Granulomata were found in three eyes of the trabeculectomy group and in three eyes of the autopsy eye group. Part of the Schlemm canal was occluded and replaced by fibrotic tissue and it became narrow in four eyes of the trabeculectomy group and three eyes of the autopsy eye group. The spaces of the trabecular meshwork appeared wide, even in the area close to granulomata and peripheral anterior synechia of the iris. Infiltration of lymphocytes, monocytes, and macrophages around the Schlemm canal was found in all eyes of the trabeculectomy group and in four eyes of the autopsy eye group. The infiltration of these cells was observed not only in the inner wall, but also in the posterior outer wall of the Schlemm canal and the collector channels. CONCLUSION: "Schlemm canalitis" is proposed from the results of inflammatory cell infiltration around the wall of the canal. The occlusion of the Schlemm canal by granulomata or fibrotic tissue replacement of the canal may play an important role in secondary open-angle glaucoma due to sarcoidosis. PMID- 12095804 TI - Correction for corneal polarization axis improves the discriminating power of scanning laser polarimetry. AB - PURPOSE: Corneal polarization axis (CPA) has been reported to affect retardation measurements obtained with scanning laser polarimetry (SLP). The purpose of this investigation was to prospectively determine whether correction for CPA improves the discriminating power of SLP for detection of mild-to-moderate glaucoma. DESIGN: Cross-sectional analysis of normal and glaucomatous eyes. METHODS: We constructed a noninvasive slit-lamp-mounted device incorporating two crossed linear polarizers and an optical retarder to measure the slow axis of corneal polarization. Complete ocular examination, standard automated perimetry, SLP imaging, and CPA measurements were performed on normal and glaucomatous eyes. One eye/subject was enrolled; if both eyes of a patient were eligible for the study, the right eye was selected. For each of the 13 SLP parameters, logistic regression was used to determine if including CPA in the model influenced the ability to discriminate between normal and glaucomatous eyes. RESULTS: Forty three normal eyes (average visual field mean defect, -0.53 +/- 1.4 dB) and 33 glaucomatous eyes (average visual field mean defect, -5.93 +/- 6.5 dB) were enrolled. CPA was significantly correlated with summary retardation parameters (average thickness and integral values) in normal (r = 0.72-0.83, P <.001 for all values) and glaucomatous eyes (r = 0.43-0.62, P =.013 to <.001). Including CPA in the model improved the ability to discriminate between normal and glaucomatous eyes for five retardation parameters quantifying retinal nerve fiber layer (RNFL) thickness (range of P values: 0.045-0.001). For inferior average thickness, area under the receiver operating characteristic (ROC) curve increased significantly (P =.002) from 0.70 to 0.78 after accounting for CPA; with a sensitivity set at 80% specificity improved from 33% to 72%. Correlations between visual field corrected pattern standard deviation and average thickness, ellipse average, superior average, and inferior average significantly increased (range of P values,.018-.001) after adjustment for CPA (r = -0.35 and -0.45, -0.38 and -0.47, -0.46 and -0.57, and -0.42 and -0.49, respectively). CONCLUSIONS: Correction for CPA significantly increases the correlation between retinal nerve fiber layer structural damage and visual function and significantly improves the discriminating power of SLP for detection of mild-to-moderate glaucoma. PMID- 12095805 TI - A prospective, randomized trial of pyrimethamine and azithromycin vs pyrimethamine and sulfadiazine for the treatment of ocular toxoplasmosis. AB - OBJECTIVE: To compare the effects of two treatment regimens, one of which included azithromycin, for the treatment of sight-threatening (near optic disk or fovea) ocular toxoplasmosis. DESIGN: Prospective, randomized open-labeled multicenter study, masked in part with regard to evaluation. METHODS: PARTICIPANTS TOTAL ENROLLMENT: 46 patients with sight-threatening ocular toxoplasmosis; pyrimethamine and azithromycin group: 24 patients; pyrimethamine and sulfadiazine group: 22 patients. INTERVENTION: Patients were randomized into two treatment regimens. Group 1 was treated with pyrimethamine and azithromycin complemented with folinic acid and the addition of prednisone from day 3. Group 2 was treated with pyrimethamine and sulfadiazine complemented with folinic acid and the addition of prednisone from day 3. Patients used study medications daily for 4 weeks. Ocular and laboratory examinations were performed at least weekly during the observation period. The study was masked in part with regard to evaluation. MAIN OUTCOME MEASURES: An assessment was made of the time to resolution of the intraocular inflammatory activity, the size of the retinochoroidal lesion, and visual acuity before and after the treatment as well as all adverse effects of treatments. RESULTS: Adverse effects were more frequent in the pyrimethamine/sulfadiazine group (P <.04), and three patients in this group had to discontinue treatment. The time to resolution of inflammatory activity, decrease in size of retinochoroidal lesions, and optimal visual acuity did not differ between the two treatment groups. The number of patients who developed recurrences during the first year after treatment was similar for both groups. CONCLUSIONS: The efficacy of the multidrug regimen with pyrimethamine and azithromycin was similar to the standard treatment with pyrimethamine and sulfadiazine. However, the frequency and severity of adverse effects was significantly lower with a regimen containing pyrimethamine and azithromycin. Multidrug therapy with the combination of pyrimethamine and azithromycin appears to be an acceptable alternative for treatment of sight-threatening ocular toxoplasmosis. PMID- 12095806 TI - The effect of long-term intermittent trimethoprim/sulfamethoxazole treatment on recurrences of toxoplasmic retinochoroiditis. AB - PURPOSE: To determine the effect of long-term intermittent trimethoprim/sulfamethoxazole treatment on recurrences of toxoplasmic retinochoroiditis. DESIGN: Prospective randomized open-labeled interventional clinical trial. METHODS: A total of 124 patients with a history of recurrent toxoplasmic retinochoroiditis were randomized to treatment with one tablet of trimethoprim (160 mg)/sulfamethoxazole (800 mg) (Bactrim F; Roche Pharmaceuticals, Rio de Janeiro, Brazil) every 3 days (61 patients) or to observation without treatment (63 patients) and were followed monthly for up to 20 consecutive months for clinical signs of disease recurrence. A recurrence was defined as a new focus of necrotizing retinochoroiditis with active inflammation either adjacent to or remote from preexisting retinochoroidal scars. RESULTS: Recurrences developed in four (6.6%) treated patients and in 15 (23.8%) controls (P =.01). Treatment was discontinued prematurely in four patients because of mild drug reactions. CONCLUSION: Long-term intermittent treatment with trimethoprim/sulfamethoxazole can reduce the rate of recurrent toxoplasmic retinochoroiditis. PMID- 12095807 TI - Clinical characteristics of serpiginous choroidopathy in North India. AB - PURPOSE: To describe clinical characteristics of serpiginous choroidopathy in the North Indian population. DESIGN: Retrospective cohort-based study. METHODS: Records of 86 patients who obtained a diagnosis of serpiginous choroidopathy at presentation or after a minimal follow-up of 2 years were retrieved. Group I included 62 patients who were diagnosed as having serpiginous choroidopathy at the initial presentation. Group II included 20 patients who initially presented as having multifocal placoid pigment epitheliopathy resembling acute posterior multifocal placoid pigment epitheliopathy (APMPPE) and had progressed over years to serpiginous choroidopathy, while group III had 4 patients with mixed presentations. Statistical analysis was done using the chi-square and Mann Whitney U test. A review of the clinical, historical, fundus photographic, and fluorescein angiographic features was performed. The demographic and clinical features, number of recurrences, final visual acuity, and final description of healed lesions were documented. RESULTS: The median age at presentation was 30 years in group I, 29 years in group II, and 31 years in group III. The disease was bilateral in five (15%) patients in group I, 12 (60%) in group II, and all four in group III. All the affected eyes in groups II and III and 58 eyes (87%) in group I had recurrences over 2 to 8 years of follow up. Compared with group I, group II was characterized by more bilaterality (P =.001), less central foveal involvement (P =.001), better final visual acuity (P =.001), the presence of healed multifocal scars (P =.001), and less subretinal fibrosis (P =.02). Four patients in group III showed mixed features. CONCLUSIONS: Serpiginous choroidopathy in our population was seen in young patients and had three distinct presentations that seemed to affect the choriocapillaris primarily. Our patients appeared to have a variation of serpiginous choroidopathy, typical of the Asian Indian population, that had some important differences from that reported in Caucasians. PMID- 12095808 TI - The role of hyperhomocysteinemia and methylenetetrahydrofolate reductase (MTHFR) C677T mutation in patients with retinal artery occlusion. AB - PURPOSE: Hyperhomocysteinemia has been established as an important risk factor for cardiovascular diseases. The aim of the present study was to investigate whether hyperhomocysteinemia and/or homozygosity for the methylenetetrahydrofolate reductase (MTHFR) C677T mutation are associated with an increased risk for retinal artery occlusion (RAO). DESIGN: Retrospective case control study. METHODS: We studied 105 consecutive patients with retinal artery occlusion and 105 age and sex-matched control subjects. Fasting plasma homocysteine levels were determined by high-performance liquid chromatography, while genotypes of the MTHFR C677T mutation were determined by polymerase chain reaction. RESULTS: Mean plasma homocysteine levels were significantly higher in patients with RAO compared with control subjects (12.2 +/- 4.8 micromol/l vs 10.3 +/- 3.4 micromol/l; P =.003). Hyperhomocysteinemia was defined by the 95th percentile of control plasma homocysteine levels as 15.8 micromol/l. Twenty (19.1%) patients with RAO exceeded this level and were therefore classified as hyperhomocysteinemic compared with 5 (4.8%) control subjects (P =.003). The odds ratio for these patients was calculated at 4.7 (95% confidence interval [CI], 1.5 15.1). Mean plasma folate levels were significantly lower in patients than in the control group (5.6 +/- 2.3 ng/ml vs. 6.3 +/- 2.5 ng/ml; P =.04). The prevalence of the homozygous genotype of methylenetetrahydrofolate reductase C677T mutation did not significantly differ between patients and controls. CONCLUSIONS: Our results suggest that hyperhomocysteinemia, but not homozygosity, for the MTHFR C677T mutation is associated with RAO. PMID- 12095809 TI - Treatment of idiopathic subfoveal choroidal neovascular lesions using photodynamic therapy with verteporfin. AB - PURPOSE: To report the effects of photodynamic therapy using verteporfin in the treatment of patients with subfoveal idiopathic choroidal vascularization (CNV). DESIGN: Interventional case series. METHODS: In a retrospective study, eight eyes of eight consecutive patients aged 55 years or younger with subfoveal idiopathic CNV treated with photodynamic therapy using verteporfin were evaluated. Visual acuity was considered to be improved if the visual angle was halved, while acuity was thought to be worse if the visual angle doubled. RESULTS: The eight patients included three men and five women with a mean (+/- SD) age of 34.6 (+/- 9.7) years (range 25-53 years). The mean follow-up time was 13.5 months. At the end of the follow-up period the visual acuity improved in five eyes (62.5%), remained unchanged in one (12.5%), and decreased in two (25%). The mean acuity improvement was 3.6 lines of Snellen acuity by the end of the follow-up period, a change that was statistically significant (P =.027, Wilcoxon signed-rank test). No patient had any complication from the treatment. CONCLUSIONS: There is no widely accepted method for treating subfoveal idiopathic CNV and all previously investigated methods have had a significant number of serious side effects. Although the follow-up time and the number of patients in this pilot study were limited, the encouraging results and lack of complications suggest that further study is indicated. PMID- 12095810 TI - Inferior limited macular translocation for subfoveal choroidal neovascularization secondary to age-related macular degeneration: 1-year visual outcome and recurrence report. AB - PURPOSE: To report the 1-year visual outcomes and incidence of persistent and recurrent choroidal neovascularization (CNV) after limited macular translocation (LMT) for subfoveal CNV in patients with age-related macular degeneration (ARMD). DESIGN: Interventional case series. METHODS: Retrospective review of 102 consecutive eyes of 101 patients that had the inferior limited macular translocation procedure for subfoveal choroidal neovascularization secondary to ARMD. The outcome measures were visual acuity at 12 months after surgery, change in visual acuity from baseline, the proportion of eyes with moderate (3 or more lines) or severe (6 or more lines) visual acuity loss, and cumulative incidence of persistent or recurrent CNV and its impact on visual acuity. Cumulative incidence was estimated using Kaplan-Meier survival analysis methods. Association between persistence and recurrence of CNV and the Snellen visual acuity recorded at each follow-up visit was evaluated using the Wilcoxon rank-sum test. RESULTS: Eighty-six (84.3%) of 102 eyes completed the 1-year follow-up. By 12 months postoperatively, 35 (40.7%) of the 86 eyes achieved visual acuity of 20/100 or better while 34 (39.5%) of the 86 eyes experienced 2 or more Snellen lines of visual improvement. In the 52 eyes with effective translocation and complete laser photocoagulation of the CNV complex with sparing of the sensory fovea, the estimated incidence of recurrence was 34.6% at 12 months (95% confidence interval of 21%-48%). Sixty-five percent of the recurrences were subfoveal and caused a decrease in visual acuity. There was a trend toward worse median change in visual acuity in eyes with persistent or recurrent CNV. CONCLUSIONS: Limited macular translocation for the treatment of subfoveal CNV secondary to ARMD is associated with improvement in visual acuity in approximately 39.5% of eyes and enables complete laser photocoagulation of the neovascular complex with sparing of the sensory macula in approximately 60.4% of eyes that complete 1 year follow-up. Persistence and recurrence of CNV are common after LMT and are important causes of vision loss. Further studies are warranted to more precisely evaluate the risks and benefits of LMT in ARMD. PMID- 12095811 TI - Effects of humidified and dry air on corneal endothelial cells during vitreal fluid-air exchange. AB - PURPOSE: To report the immediate anatomic and functional alterations in corneal endothelial cells following use of humidified air and dry air during vitreal fluid-air exchange in rabbits. DESIGN: Experimental study. METHODS: Rabbits undergoing pars plana vitrectomy and lensectomy were perfused with either dry or humidified air during fluid-air exchange for designated durations. Three different experiments were performed. First, control and experimental corneas were examined by scanning electron microscopy (SEM). Second, corneas were stained with Phalloidin-FITC and examined by fluorescein microscopy. Finally, third, transendothelial permeability for carboxyfluorescein was determined using a diffusion chamber. RESULTS: While different from the corneal endothelial cells, those cells exposed to humidified air were less stressed than cells exposed to dry air by SEM. Actin cytoskeleton was found highly disorganized with dry air exposure. Humidified air maintained the normal actin cytoskeleton throughout the 20 minutes of fluid-air exchange. Paracellular carboxyfluorescein leakage was significantly higher in dry air insufflated eyes compared with that of the humidified air after 5, 10, and 20 minutes of fluid-air exchange (P =.002, P =.004, and P =.002, respectively). CONCLUSIONS: Dry air stress during fluid-air exchange causes significant immediate alterations in monolayer appearance, actin cytoskeleton, and barrier function of corneal endothelium in aphakic rabbit eyes. Use of humidified air largely prevents the alterations in monolayer appearance, actin cytoskeleton, and barrier function of corneal endothelial cells. PMID- 12095812 TI - Long-term prognosis in patients with vasculopathic sixth nerve palsy. AB - PURPOSE: To better define the long-term prognosis in patients with a vasculopathic sixth nerve palsy (6NP), specifically addressing the degree of recovery and incidence of recurrent similar episodes. DESIGN: Observational case series. METHODS: Retrospective chart review. SETTING: An outpatient neuroophthalmic practice. STUDY POPULATION: Patients with one or more vascular risk factors and an acute, isolated 6NP that spontaneously recovered. OBSERVATION PROCEDURE: Information regarding resolution of the 6NP, subsequent vascular events and recurrent ocular motor nerve palsy was obtained from chart review of follow-up clinic visits, mailed questionnaires and telephone interviews. The duration of follow-up ranged from 2 to 13 years. MAIN OUTCOME MEASURES: Resolution of 6NP (complete or incomplete) and incidence of recurrent ocular motor nerve palsy. RESULTS: Fifty-nine patients were identified with a mean age of 65.3 years +/- 11.6 (range 34-90 years). Fifty-one patients (86%) experienced complete resolution of their first episode of vasculopathic 6NP and eight patients (14%) had incomplete resolution. A subsequent episode of ocular motor mononeuropathy occurred in 18 of 59 (31%) patients. The number of recurrences ranged from one (in 14 patients) to four (in one patient). There was no association between any risk factor and recurrence of ocular motor nerve palsy. Similarly, incomplete resolution of the vasculopathic 6NP was not associated with any risk factor. CONCLUSIONS: Patients with a vasculopathic 6NP usually have complete resolution of their ophthalmoplegia, but nearly one third of patients in our study later experienced at least one episode of recurrent vasculopathic ocular motor nerve palsy. PMID- 12095813 TI - Dural carotid cavernous fistula: definitive endovascular management and long-term follow-up. AB - PURPOSE: To describe the endovascular treatment and clinical outcome in patients with indirect carotid cavernous fistulas (CCFs) over a 15-year period. To our knowledge, this is the largest series in the medical literature. DESIGN: Interventional case series. METHODS: A retrospective evaluation of 135 consecutive patients who underwent examination and treatment for indirect CCF was performed. Patients received independent evaluations by ophthalmologists, neurologists, or neuro-ophthalmologists before, during, and after endovascular treatment. Patients initially received noninvasive imaging followed by cerebral arteriography for definitive diagnosis and stratification by angiographic risk factors. Endovascular treatment was performed in 133 (98%) patients and clinical follow-up was achieved in 135 (100%) patients on an average of 56 +/- 4.3 months (range: 2 months-14 years). Angiographic follow-up was performed in 72 (54%) patients with ongoing symptoms or a history of fistula with high-risk angiographic features. Arteriographic cure with long-term clinical outcome is summarized by modified Rankin scale (mRS) and Barthel index (BI). RESULTS: At a mean follow-up of 56 months, 121 (90%) patients were clinically cured. At latest clinical follow-up, 131 (97%) patients showed good recovery (mRS, 1-2; BI 90 100), one (1%) had moderate disability (mRS, 3; BI, 50-60), and three (2%) (mRS, 4; BI, 40-50) were severely disabled. Procedure-related permanent morbidity was 2.3%. There was no operative mortality. CONCLUSIONS: With the observed favorable outcomes and low rate of procedural morbidity in this patient population with long-term angiographic and clinical follow-up, endovascular therapy should be the primary treatment for patients with indirect (dural) fistulas of the cavernous sinus. PMID- 12095814 TI - Relation between optical coherence tomography and optical pachymetry measurements of corneal swelling induced by hypoxia. AB - PURPOSE: To determine the relation between optical coherence tomography (OCT) and optical pachymetry (OP) measurements of corneal swelling induced by hypoxia. DESIGN: Experimental study. METHODS: One randomly selected eye of 20 noncontact lens wearers (10 males and 10 females, age 35.6 +/- 9.6 years) was patched during 3 hours of soft contact lens (SCL) wear while the contralateral eye acted as control. Central corneal thickness of both eyes was measured before and after SCL wear using OCT and OP in randomized order. RESULTS: Baseline central corneal thickness was 523.6 +/- 33.0 microm (mean +/- standard deviation [SD]) measured with OCT and 490.6 +/- 25.5 microm with OP. Immediately after contact lens removal, corneal thickness measured with OCT increased by 13.8 +/- 2.3% compared with 12.1 +/- 1.8% (paired t test: P <.001) measured with OP. Thereafter, corneal thickness decreased at the rate of 5.6% per hour for OCT and 5.4% per hour for OP. The difference in thickness between instruments before lens insertion, which was 33 microm compared with the difference after lens removal (edematous cornea), which ranged from 46 to 41 microm. The difference between instruments decreased during the corneal deswelling period after lens removal. The correlation coefficient between OCT and OP was 0.914 before lens insertion and 0.932 after lens removal. CONCLUSION: This study has demonstrated the difference of corneal thickness measured with OCT and OP. Although both instruments are correlated highly in all conditions tested, OCT may overestimate corneal thickness in normal and edematous corneas. PMID- 12095815 TI - Treatment of juxtafoveal and extrafoveal choroidal neovascularization in the era of photodynamic therapy with verteporfin. PMID- 12095816 TI - An update on current practices in the management of ocular toxoplasmosis. AB - PURPOSE: To update information that was published by the AMERICAN JOURNAL OF OPHTHALMOLOGY in 1991 about treatment practices for ocular toxoplasmosis by uveitis specialists. DESIGN: Physician survey. METHODS: A written questionnaire was distributed to all physician-members (n = 147) of the American Uveitis Society. The questionnaire was modeled after a similar device used to survey uveitis specialists in 1991. Information contained on 96 returned questionnaires was tabulated. RESULTS: Among 79 respondents who evaluate and manage patients with ocular toxoplasmosis, 15% treat all cases regardless of clinical findings (in contrast to 6% in 1991). The major indications for treatment among other respondents were severe inflammatory responses and proximity of retinal lesions to the fovea and optic disk. The majority of clinical factors considered in five categories (vision, lesion location, lesion size, lesion characteristics, and vitreous inflammatory reaction) were identified to be relative or absolute indications for treatment by a greater proportion of respondents in the current survey than in the 1991 survey. A total of nine drugs (or commercially available combinations) were used in 24 different regimens as treatments of choice for typical cases of recurrent toxoplasmic retinochoroiditis, with the combination of pyrimethamine, sulfadiazine, and prednisone being the most commonly used regimen (29% of respondents). CONCLUSIONS: Uveitis specialists appear to be more likely to treat patients with ocular toxoplasmosis in 2001 than in 1991. Although the majority of survey respondents adhere to a traditional approach to the management of toxoplasmic retinochoroiditis (a discrete course of systemic drug treatment during active disease using multiple antiparasitic drugs with or without corticosteroids), there is still no consensus regarding the choice of antiparasitic agents for treatment regimens. Survey results provide useful information for treating physicians and for clinical investigators interested in therapy. PMID- 12095817 TI - Effect of tear film break-up on higher-order aberrations measured with wavefront sensor. AB - PURPOSE: To investigate whether optical wavefront aberrations vary with tear film break-up. DESIGN: Observational case series. METHODS: Higher-order aberrations were examined for 20 eyes of 20 normal subjects with a Hartmann-Shack wavefront sensor before and after tear film break-up. RESULTS: Higher-order aberrations for photopic vision (central 4 mm diameter) after tear film break-up increased 1.44 fold compared to higher-order aberrations before tear film break-up (P =.001, paired t-test). Higher-order aberrations after tear film break-up for scotopic vision (central 6 mm diameter) were also 1.23 times higher than those before break-up (P =.005, paired t-test). CONCLUSION: Wavefront sensing enabled us to evaluate the induced irregular astigmatism caused by tear film break-up quantitatively. Wavefront aberrations should be measured carefully to avoid the effects of tear film break-up, especially in wavefront-guided refractive surgery. PMID- 12095818 TI - Conjunctival mycosis masquerading as melanoma. AB - PURPOSE: To report a patient with a melanotic lesion of the conjunctiva that mimicked melanoma but proved to be a fungal infection on biopsy. DESIGN: Interventional case report. METHODS: A 75-year-old white man was examined for a 3 month history of a deeply pigmented lesion of the conjunctiva at the limbus of the right eye that closely resembled conjunctival melanoma. The lesion was completely excised. RESULTS: Histologic examination revealed fungal elements without evidence of tumor. CONCLUSION: Although rare, fungi should be included in the differential diagnosis of pigmented lesions of the conjunctiva. PMID- 12095819 TI - Intraocular caterpillar setae without subsequent vitritis or iridocyclitis. AB - PURPOSE: To report a case of caterpillar setae embedded in the corneal stroma and inferotemporal retina with minimal inflammation. DESIGN: Observational case report. METHODS: A 4-year-old boy developed a red eye after playing with a caterpillar. He was placed on topical tobramycin/dexamethasone and referred for evaluation of embedded setae in his conjunctiva, cornea, iris, and retina. Examination revealed no iridocyclitis or vitritis. RESULTS: Because of the lack of intraocular inflammatory response, no invasive intervention was conducted to remove or destroy the setae and he was tapered off the tobramycin/dexamethasone. At 4-month follow-up he remained asymptomatic with the setae still present in both his cornea and retina. In addition, vitreous membranes had formed in the immediate vicinity of the intraretinal setae. CONCLUSION: Intraretinal and corneal setae can be embedded with minimal inflammation and can be tolerated without need for surgical intervention. PMID- 12095820 TI - Donor-to-host transmission of Candida albicans after corneal transplantation. AB - PURPOSE: To report donor-to-host transmission of Candida albicans after penetrating keratoplasty. DESIGN: Interventional case report. METHODS: A 15-year old boy who underwent penetrating keratoplasty for keratoconus with donor tissue from a drowning victim developed keratitis and a lenticular abscess 26 days postoperatively. RESULTS: Candida albicans was cultured from the donor rim and the recipient cornea. Antifungal sensitivity profiles were identical for the two isolates. DNA profiles were identical for both isolates, confirming the donor as the source of the infection. CONCLUSION: This case demonstrates the value of routine culture of corneal donor rims and the advisability of close follow-up and possible antifungal prophylaxis when donor rims are positive for fungus. PMID- 12095821 TI - Refractory intraocular pressure increase after phakic posterior chamber intraocular lens implantation. AB - PURPOSE: To describe a complication resulting from the implantation of a posterior chamber intraocular lens in a phakic eye. DESIGN: Interventional case report. METHODS: We examined a 37-year-old woman with severe myopia who had implantation of a phakic posterior chamber intraocular lens. RESULTS: The patient developed pigmentary glaucoma with refractory increase in intraocular pressure, despitemedical therapy and intraocular lens removal. Trabeculectomy was required to reduce the pressure. CONCLUSION: This case demonstrates that pigmentary glaucoma secondary to implantation of a phakic posterior chamber intraocular lens can lead to filtering surgery to decrease intraocular pressure. PMID- 12095822 TI - Retinal visualization in an eye with corneal crystals using indocyanine green videoangiography. AB - PURPOSE: To report a patient in whom clear imaging of the retina, impossible with conventional methods, was obtained using indocyanine green (ICG) videoangiography. DESIGN: Interventional case report. METHODS: A 31-year-old patient with nephropathic cystinosis and complaints of decreased vision in the left eye underwent a complete ophthalmologic evaluation, including conventional photography, fluorescein angiography, and ICG videoangiography. RESULTS: With conventional methods (direct and indirect ophthalmoscopy, photography, and fluorescein angiography), the view of the left retina was obscured by densely packed corneal cystine crystals. During ICG angiography, clear imaging of the retina was obtained with near-infrared illumination. CONCLUSION: Indocyanine green videoangiography can be a useful tool in cases with cystinosis and other corneal opacities, where visualization and imaging of the retina are important but impossible with conventional methods. PMID- 12095823 TI - Sequential multifocal electroretinogram findings in a case of Purtscher-like retinopathy. AB - PURPOSE: To report the multifocal electroretinographic findings of a patient with pancreatitis-associated Purtscher-like retinopathy. DESIGN: Observational case report. METHODS: A 20-year-old man with a history of alcohol abuse and acute pancreatitis underwent multifocal electroretinography (mfERG; 103-scaled hexagons, 8-minute recording time) at 1 week, 4 weeks, and 6 months after the onset of the retinopathy. RESULTS: At 1 week, the patient had extensive cotton- wool spots in the posterior pole bilaterally, and visual acuity was significantly reduced. Both the A-waves and B-waves of the mfERG were depressed in the corresponding areas. At 4 weeks, neither visual acuity nor mfERG showed improvement, although the cotton-wool spots had resolved. Interestingly, at 6 months, visual acuity had improved significantly in the left eye, consistent with increased ratios of mfERG responses for the central area. CONCLUSION: There was no selective reduction of b-waves of the mfERG as anticipated in isolated inner retinal pathology. In this case, both the A-waves and B-waves were reduced, suggesting damage to both outer and inner retinal layers. PMID- 12095824 TI - Autoantibodies to transducin in a patient with melanoma-associated retinopathy. AB - PURPOSE: To report novel immunoreactivity in a patient with melanoma-associated retinopathy. DESIGN: Retrospective case report and experimental study. METHODS: A 32-year-old woman with a history of metastatic melanoma presented with bilateral decreased visual acuity. Electroretinography, Goldmann perimetry, immunohistochemistry, and Western blotting of her serum were performed. RESULTS: Electroretinography showed a "negative" B-wave. Paracentral and central scotomas were observed on Goldmann perimetry. Antibodies to a retinal transducin were demonstrated by Western blotting. No immunoreactivity to retinal bipolar cells was detected by immunohistochemistry. CONCLUSION: Melanoma-associated retinopathy can be related to a variety of antiretinal antibodies. Recognition of transducin, a novel melanoma-associated retinopathy antigen, may be important for identifying and treating patients with night blindness and melanoma. PMID- 12095825 TI - Rheumatoid hyperviscosity syndrome: reversibility of microvascular abnormalities after treatment. AB - PURPOSE: To report a case of rheumatoid hyperviscosity syndrome involving both retinal and choroidal circulation that resolved after treatment. DESIGN: Interventional case report. METHODS: A 58-year-old woman with clinical and serologic evidence of an inflammatory connective tissue disease without any visual complaints was referred for a funduscopic evaluation. RESULTS: Funduscopic examination revealed marked dilation and beading of the venous system, microaneurysms, and telangiectatic capillary beds in the posterior pole. Fluorescein angiography disclosed delayed choroidal filling, prolonged arteriovenous transit time, and areas of capillary nonperfusion. These findings were accompanied by a severe polyclonal hypergammaglobulinemia and a 10-fold increase in serum viscosity. The ocular findings were reversible after plasmapheresis and steroid treatment. CONCLUSION: Rheumatoid hyperviscosity syndrome can involve both retinal and choroidal circulation. The prominent microvasculopathy is reversible after appropriate treatment. PMID- 12095826 TI - Retinal and choroidal vascular occlusion after posterior sub-tenon triamcinolone injection. AB - PURPOSE: To report a case of retinal and choroidal vascular occlusion occurring as a complication after posterior sub-Tenon triamcinolone injection for treatment of uveitic cystoid macular edema. DESIGN: Interventional case report. METHODS: Retrospective study. A 32-year-old woman with uveitis and cystoid macular edema underwent a right posterior sub-Tenon injection of triamcinolone (40 mg/ml, 1 ml total) through a superotemporal approach after topical anesthesia. After the procedure, the patient experienced severe eye pain, orbital ecchymosis, and globe proptosis consistent with retrobulbar hemorrhage. RESULTS: Dilated fundus examination of the right eye (OD) demonstrated multiple intraretinal hemorrhages with particulate white emboli occluding the retinal and choroidal vessels. Visual acuity was no light perception. Ocular massage and hypotensive therapy was initiated for an intraocular pressure of 50 mm Hg. Canthotomy and cantholysis were performed. A total of 39 months post-incident, her visual acuity improved to 20/100. CONCLUSION: Posterior sub-Tenon triamcinolone injection can rarely result in retinal and choroidal occlusion. Immediate intervention may preserve limited visual acuity. PMID- 12095827 TI - Choroidal neovascularization in primary hyperoxaluria. AB - PURPOSE: To report a case of primary hyperoxaluria in which choroidal neovascularization developed bilaterally. DESIGN: Observational case report. METHODS: A 22-year-old man with a history of type I primary hyperoxaluria complained of a slow but progressive loss of vision in both eyes for the preceding 8 months. The clinical, fluorescein, and ocular coherence tomography findings are reported. RESULTS: Fluorescein angiography confirmed the presence of choroidal neovascularization in both eyes at the edge of previous macular scars. Ocular coherence tomography scans were obtained to better characterize the clinical pathology. CONCLUSION: In a patient with type 1 primary hyperoxaluria, the presence of choroidal neovascularization in both eyes outside but adjacent to an area of previous macular scarring is reported. These findings are in harmony with the assumption that mechanical factors from the oxalate deposition promote choroidal neovascularization. PMID- 12095828 TI - Photodynamic therapy with verteporfin for subfoveal choroidal neovascularization in Vogt-Koyanagi-Harada syndrome. AB - PURPOSE: To assess the role of photodynamic therapy using verteporfin in the treatment of subfoveal choroidal neovascularization in Vogt-Koyanagi-Harada syndrome. DESIGN: Interventional case report. METHODS: A 9-year-old patient with subfoveal choroidal neovascularization received a single photodynamic therapy with verteporfin session (one eye) and was prospectively followed with fluorescein angiography. RESULTS: A complete regression of the lesion was achieved within 1 week after treatment. Visual acuity improved from 20/800 to 20/320 by 6 months of follow-up. Fluorescein angiography disclosed unexpected retinal pigment epithelium alteration within the treatment area. CONCLUSION: Although regression of the choroidal neovascularization occurred, unpredicted findings involving normal retina in the vicinity of the lesion suggest that further studies are required to assess the clinical value of this treatment for subfoveal choroidal neovascularization in Vogt-Koyanagi-Harada syndrome. PMID- 12095829 TI - Severe hypotony after macular translocation surgery with 360-degree retinotomy. AB - PURPOSE: To report a case of severe hypotony after macular translocation with 360 degree retinotomy. DESIGN: Interventional case report. METHODS: A 50-year-old woman with myopic neovascular maculopathy underwent macular translocation with 360-degree retinotomy in her left eye. RESULTS: After the second procedure of silicone oil removal, severe hypotony developed. No clear sign of leakage was found. Pure perfluoropropane gas tamponade was then performed, which resulted in temporal resolution of severe hypotony, but the hypotony recurred as the gas bubble was absorbed. Ten weeks after the second surgery, the hypotonous eye was refilled with silicone oil. No apparent cyclitic membrane was observed intraoperatively. After this procedure, the choroidal and retinal folds regressed; intraocular pressure has been between 5 and 7 mm Hg for more than 4 months thereafter. CONCLUSION: Severe hypotony can occur as a complication of otherwise uneventful macular translocation with 360-degree retinotomy. PMID- 12095830 TI - Bilateral optic disk edema and blindness as initial presentation of acute lymphocytic leukemia. AB - PURPOSE: To report bilateral optic disk edema and blindness as the unusual initial presentation of acute lymphocytic leukemia (ALL) in an adult. METHODS: A 19-year-old man presented with a history of headaches, back pain, and 10 days of worsening vision that progressed to blindness. Ocular examination revealed light perception acuity in the right eye and no light perception in the left eye. Fundus examination revealed bilateral profound optic disk edema, tortuous vessels, and retinal hemorrhages. Acute lymphocytic leukemia was diagnosed with complete blood count and bone marrow biopsy. Head computed tomography and magnetic resonance imaging, were normal. Lumbar puncture revealed normal opening pressure. Ocular ultrasonography showed bilateral optic nerve enlargement. DESIGN: Interventional case report and literature review. RESULTS: The presumptive diagnosis of leukemic infiltration of the optic nerves was made, and urgent radiotherapy, intrathecal methotrexate, and intravenous daunorubicin were instituted. Visual acuity improved to hand motions in the right eye. CONCLUSIONS: Acute lymphocytic leukemia can rarely present in adults as visual changes due to leukemic optic nerve infiltration. Radiation treatment should be considered as an urgent treatment modality for this rare condition. PMID- 12095831 TI - The detection of a relative afferent pupillary defect. AB - PURPOSE: To describe a simple technique that detects a subtle relative afferent pupillary defect. DESIGN: Observational report. METHODS: With the patient seated at a slit lamp and the slit illumination turned off, a transilluminator light is swung back and forth between the eyes. The examiner observes the direct pupil light response and the consensual light response under slit-lamp magnification. CONCLUSION: Using slit-lamp magnification, the examiner may observe small amplitudes of pupillary movement in the evaluation of a relative afferent pupillary defect. PMID- 12095832 TI - Discordant pursuit asymmetry and esotropia in monozygous twins. AB - PURPOSE: To report the relationship between infantile esotropia and asymmetry of monocular pursuit tracking. DESIGN: Observational family case series. METHODS: A twin pair discordant for infantile esotropia was confirmed by DNA analysis to be monozygous at 99.95% probability. Digitally sampled electro-oculographic recordings of monocular horizontal pursuit eye movements were performed for the twins and their orthophoric father. RESULTS: Mean directional asymmetry for right and left eyes was 31% and 5% for the esotropic twin, 10% and 8% for the orthophoric twin, and 5% and 6% for the father. The asymmetry was statistically significant only for the deviating right eye of the esotropic twin (P = 0.0217). CONCLUSION: In this family, monocular pursuit asymmetry is probably not a heritable predisposition to infantile esotropia, but a consequence of early strabismus. Discordance of infantile esotropia and pursuit asymmetry in monozygous twins implicates environmental in addition to genetic causes. PMID- 12095834 TI - Skew deviation and inferior oblique palsy. PMID- 12095833 TI - Loss of fragile histidine triad (FHIT) expression and microsatellite instability in periocular sebaceous gland carcinoma in patients with Muir-Torre syndrome. AB - PURPOSE: To report fragile histidine triad expression and microsatellite instability in periocular sebaceous gland carcinoma. DESIGN: Interventional case series. METHODS: Biopsy specimens of periocular sebaceous gland carcinoma obtained from six patients (mean age, 60 +/- 17 years; range, 38 to 83 years, 5 male, 1 female) with Muir-Torre syndrome and histopathologically proven sebaceous gland carcinoma were studied immunohistochemically for the presence of fragile histidine triad protein. Polymerase chain reaction-based analysis of the markers BAT25, BAT26, D2S123, D5S346, and D17S250 was performed for microsatellite instability in tumorous and matching normal tissues. RESULTS: Fragile histidine triad protein was detectable in the sebaceous gland carcinoma from one patient with microsatellite instability. It was not detectable in sebaceous gland carcinoma specimens from five patients without any evidence of microsatellite instability. CONCLUSION: Inactivation of fragile histidine triad tumor suppressor gene or inactivation of the mismatch-repair system resulting in microsatellite instability may contribute to the development of periocular sebaceous gland carcinoma in Muir-Torre syndrome. PMID- 12095836 TI - Predictive value of a 1-day, 1-eye trial on the 1-month ocular hypotensive response of latanoprost 0.005%. PMID- 12095838 TI - Retinal pigment epithelial changes after macular hole surgery with indocyanine green-assisted internal limiting membrane peeling. PMID- 12095854 TI - Comparative cardiovascular development: improving the conceptual framework. AB - Immature vertebrates-either as an embryo in an egg, as free-living larva, or as an in utero fetus, are clearly not just small versions of adults. Their cardiovascular physiology (and doubtlessly other aspects of physiology) differs from that of adults both qualitatively and quantitatively. Yet, comparative cardiovascular physiologists have been relatively conservative in constructing a new (or at least modified) conceptual framework for the understanding of developmental cardiovascular physiology. We recommend that this framework rely less on the established cardiovascular truisms for adult cardiovascular physiology that are proving to be less useful and in instances even inaccurate for interpreting development of the heart and vasculature. We have suggested that three methodologies in particular be incorporated to a greater extent in studies of comparative cardiovascular development: (a) emphasis on multivariate approaches; (b) differentiation between absolute (extrinsic) and relative (intrinsic) time for development, and; (c) employment of time lines for both intra- and interspecific comparisons of the ontogeny of cardiovascular processes. While certainly none of these approaches are novel and others have previously dwelt at length on their importance in other contexts, we feel that the emerging framework for investigating cardiovascular physiological development would benefit from incorporating these and other approaches into experimental design as well as data analysis. Failing to do so results in a heavy dependence on analytical approaches typically used for adults, and thus under-appreciates the novelty and complexity of the developing vertebrate cardiovascular system. PMID- 12095855 TI - Development of cardiac rhythms in birds. AB - Heart rate (HR) in avian embryos developing inside an eggshell has been measured by various means while maintaining adequate gas exchange through the eggshell. This is an important requirement in order to avoid adverse effects of impeding gas exchange on the cardiac rhythms of developing embryos. The present report is a review of our ontogenetic study on embryonic HR, which was measured with fulfillment of the above requirement and also hatchling HR measured non invasively. Firstly, we reviewed measurements of daily changes (developmental patterns) in embryonic mean heart rate (MHR), which were determined from a short term measurement of HR once a day, in 34 species of altricial and precocial birds. The allometric relationship between the MHR during pipping in altricial birds and their fresh egg masses was the same as that between the MHR at 80% of incubation duration and fresh egg masses in pre-cocial birds. Secondly, we presented the developmental patterns of MHR in chick embryos and hatchlings, which were determined from long-term, continuous measurement of HR before, during and after hatching. The ultradian and circadian rhythms of HR were clearly shown in embryos and hatchlings, respectively. Thirdly, we summarized instantaneous HR fluctuations: HR variability and HR irregularities, in chick embryos and hatchlings. The distinctive patterns were shown in pre-pipped and pipped embryos and newly hatched chicks, individually, which were partly related to autonomic nervous functions and physiological functions. PMID- 12095856 TI - Ontogeny of excitation-contraction coupling in the mammalian heart. AB - The neonate mammalian heart is phenotypically different from the adult heart in many respects. Understanding these phenotypic differences are a fundamental component of understanding the mechanisms of congenital heart disease and its treatment. Differences in excitation-contraction (E-C) coupling of the neonatal heart from that of the adult include less reliance on intercellular sources of Ca(2+) such as that from sarcoplasmic reticulum (SR). Electron micrographs indicate that these immature cardiomyocytes lack transverse tubules and the SR is sparse. This paper focuses on the changes in the phenotype of E-C coupling during ontogeny in the mammalian heart and the molecular mechanisms underlying these changes. PMID- 12095857 TI - Perspectives on mammalian cardiovascular aging: humans to molecules. AB - Age-related changes in cardiovascular function and structure in healthy adult volunteer community dwelling subjects (from 20 to 85 years) is remarkable for changes in pump function [impaired left ventricular (LV) ejection reserve capacity manifest by a reduced ejection fraction and accompanied by diminished cardioacceleration, LV dilation at end diastole and an altered diastolic filling pattern] and increased vascular afterloading. There is also evidence for a reduction in the number of cardiac myocytes with advancing age. Subcellular changes with aging (best understood in rodents) include certain regulatory factors of excitation-contraction-relaxation coupling (i.e. calcium handling), modulation by adrenergic receptor (AR) stimulation, and changes in the generation and sensitivity to the damaging effects of ROS. Coordinated changes in gene expression and/or protein function with aging result in a prolonged action potential (AP), Ca(i) transient, and contraction. L-type Ca(2+) current (I(Ca)) inactivates more slowly, and outwardly-directed K(+) currents are reduced, and likely contribute to AP-prolongation. The rate of Ca(2+) sequestration by the sarcoplasmic reticulum (SR) decreases in the senescent myocardium, in part underlying the prolonged Ca(i) transient. An age-associated reduction in transcription of the SERCA2 gene, coding for the SR Ca(2+) pump, accounts in part for a decrease in the SR pump site density. The contractile response to both beta(1)-AR and beta(2)-AR stimulation diminishes with aging due to decreased adrenergic augmentation of I(Ca), and thus the Ca(i) transient, in senescent vs. young hearts. The age-associated reduction in the postsynaptic response of myocardial cells to beta(1)-AR stimulation appears to be due to multiple changes in molecular and biochemical receptor coupling and post-receptor mechanisms. An increased basal production of ROS is paralleled by increased ROS-sensitivity, markers of chronic ROS damage and mitochondrial functional decline. Overall, these changes lead to a diminished (but not necessarily exhausted) capacity of the heart to adapt to physiological or pathological stress with advancing age. PMID- 12095858 TI - Coronary arteriosclerosis in salmon: growing old or growing fast? AB - A review is presented of what we know and what we suspect regarding the formation of coronary arteriosclerotic lesions in salmonids. Coronary lesions are a fact of life for both Atlantic and Pacific species of migrating salmon. Severe forms of lesions, usually restricted to the main coronary artery, are typically found in the majority of a salmon population when they are spawning. Vascular injury to the coronary artery, as a result of the bulbus arteriosus being excessively distended, is proposed as an initiating mechanism for coronary lesion formation, possibly explaining why severe lesions are restricted primarily to the main coronary artery. Evidence is presented that coronary arteriosclerosis in salmonids develops in immature fish, well before maturation, and progresses with age. Growth and growth rate are implicated in lesion progression. A faster growth rate could produce a more stressful life style, which in turn initiates more coronary vascular injury. Dietary factors, especially polyunsaturated fatty acids (and their metabolites), can significantly stimulate vascular smooth muscle proliferation in the salmon coronary artery, but a possible linkage to the progression of coronary lesions has yet to be studied. Whether coronary lesions negatively impact blood flow to the salmon heart has not been properly studied. Nevertheless, the coronary blood supply to the heart has functional importance when salmon exercise and the coronary flow reserve may be reached when fish swim under mild hypoxic conditions. If coronary arterial lesions do adversely affect blood flow to the heart, the selective effects would be most prominent in years when upstream migration conditions are particularly severe. PMID- 12095860 TI - Climate variations and the physiological basis of temperature dependent biogeography: systemic to molecular hierarchy of thermal tolerance in animals. AB - The physiological mechanisms limiting and adjusting cold and heat tolerance have regained interest in the light of global warming and associated shifts in the geographical distribution of ectothermic animals. Recent comparative studies, largely carried out on marine ectotherms, indicate that the processes and limits of thermal tolerance are linked with the adjustment of aerobic scope and capacity of the whole animal as a crucial step in thermal adaptation on top of parallel adjustments at the molecular or membrane level. In accordance with Shelford's law of tolerance decreasing whole animal aerobic scope characterises the onset of thermal limitation at low and high pejus thresholds (pejus=getting worse). The drop in aerobic scope of an animal indicated by falling oxygen levels in the body fluids and or the progressively limited capacity of circulatory and ventilatory mechanisms. At high temperatures, excessive oxygen demand causes insufficient oxygen levels in the body fluids, whereas at low temperatures the aerobic capacity of mitochondria may become limiting for ventilation and circulation. Further cooling or warming beyond these limits leads to low or high critical threshold temperatures (T(c)) where aerobic scope disappears and transition to an anaerobic mode of mitochondrial metabolism and progressive insufficiency of cellular energy levels occurs. The adjustments of mitochondrial densities and their functional properties appear as a critical process in defining and shifting thermal tolerance windows. The finding of an oxygen limited thermal tolerance owing to loss of aerobic scope is in line with Taylor's and Weibel's concept of symmorphosis, which implies that excess capacity of any component of the oxygen delivery system is avoided. The present study suggests that the capacity of oxygen delivery is set to a level just sufficient to meet maximum oxygen demand between the average highs and lows of environmental temperatures. At more extreme temperatures only time limited passive survival is supported by anaerobic metabolism or the protection of molecular functions by heat shock proteins and antioxidative defence. As a corollary, the first line of thermal sensitivity is due to capacity limitations at a high level of organisational complexity, i.e. the integrated function of the oxygen delivery system, before individual, molecular or membrane functions become disturbed. These interpretations are in line with the more general consideration that, as a result of the high level of complexity of metazoan organisms compared with simple eukaryotes and then prokaryotes, thermal tolerance is reduced in metazoans. A similar sequence of sensitivities prevails within the metazoan organism, with the highest sensitivity at the organismic level and wider tolerance windows at lower levels of complexity. However, the situation is different in that loss in aerobic scope and progressive hypoxia at the organismic level define the onset of thermal limitation which then transfers to lower hierarchical levels and causes cellular and molecular disturbances. Oxygen limitation contributes to oxidative stress and, finally, denaturation or malfunction of molecular repair, e.g. during suspension of protein synthesis. The sequence of thermal tolerance limits turns into a hierarchy, ranging from systemic to cellular to molecular levels. PMID- 12095861 TI - Beating the cold: the functional evolution of troponin C in teleost fish. AB - The sensitivity of the cardiac myocyte contractile element for Ca(2+) decreases with temperature. As myocyte contractility is regulated by changes in cytosolic [Ca(2+)], this desensitizing effect represents a challenge for temperate fish such as the rainbow trout, Oncorhynchus mykiss, living in environments where temperatures are low and variable. To allow cardiac function in a temperate environment it is thought that the comparatively high Ca(2+) sensitivity of trout cardiac myocytes compensates for the effects of low temperature on myocyte contractility. The high Ca(2+) sensitivity of the trout myocyte is due, at least in part, to changes in the amino acid sequence of the thin filament protein, cardiac troponin C (cTnC). cTnC is the Ca(2+)-activated switch that triggers myocyte contraction. The isoform of cTnC cloned from trout ventricle (ScTnC) is 92% identical to mammalian cTnC (McTnC) and is significantly more sensitive to Ca(2+). This result suggests that ScTnC has evolved in trout to allow cardiac function at low temperatures. cTnC also appears to play a role in maintaining cardiac function when temperatures change. Increasing myofibrillar pH according to alpha-stat regulation, as would occur when temperature decreases, increases Ca(2+) sensitivity. A similar increase in pH also sensitizes cTnC to Ca(2+). ScTnC therefore appears critical in maintaining cardiac function in trout at low temperatures as well as during changes in temperature. PMID- 12095862 TI - Temperature and angiogenesis: the possible role of mechanical factors in capillary growth. AB - This review examines the effect of prolonged cold exposure on muscle capillary supply in mammals and fishes. In rats and hamsters, the response to a simulated onset of winter is to conserve the microcirculation and maintain a constant capillary to fibre ratio (C:F), implying either an unaltered vacular bed or angiogenesis matched by muscle hyperplasia, while chronic acclimation to low environmental temperature induces a variable degree of muscle atrophy, which in turn increases capillary density (CD). In striped bass and rainbow trout, cold induced angiogenesis results in an increase in C:F, but also a cold-induced fibre hypertrophy that is accompanied by a powerful angiogenic response such that CD is much less sensitive to changes in fibre size. Endothelial cells can act as mechanotransducers such that angiogenesis may be initiated by changes in their physical environment. It is hypothesised that in mammals, the metabolic consequences of cold exposure increases the luminal shear stress, while in fishes the stimulus for angiogenesis is abluminal stretch following an increase in fibre size. PMID- 12095863 TI - Metabolic cold adaptation of polar fish based on measurements of aerobic oxygen consumption: fact or artefact? Artefact! AB - Whether metabolic cold adaptation in polar fish, based on measurements of aerobic standard metabolic rate, is a fact or an artefact has been a dispute since Holeton asked the question in 1974. So far polar fish had been considered to be metabolically cold adapted because they were reported to have a considerably elevated resting oxygen consumption, or standard metabolic rate, compared with oxygen consumption values of tropical or temperate fish extrapolated to similar low polar temperatures. Recent experiments on arctic and Antarctic fish, however, do not show elevated resting aerobic oxygen consumption values, or standard metabolic rate, and hence it is concluded that that metabolic cold adaptation in the traditional sense is an artefact. PMID- 12095864 TI - Cardiorespiratory performance in salmonids during exercise at high temperature: insights into cardiovascular design limitations in fishes. AB - Studies in the laboratory with salmonids and now in the field with wild salmon clearly show that critical swimming performance has an optimum temperature. This temperature optimum is coincident with maximum aerobic scope and maximum cardiac scope. At a temperature that is higher than this optimum, however, whole animal performance declines abruptly. Evidence is presented here to suggest that this is directly associated with a decline in cardiac scope which limits oxygen supply to tissues. It is further suggested that the decline in maximum cardiac performance could reflect problems with the heart's own oxygen supply. The reasoning behind this suggestion is that, at temperatures at or below the optimum and probably because of a limitation on oxygen diffusion in skeletal muscle during exercise, venous oxygen does not fall below a threshold level during exercise, and so the heart receives just enough oxygen for its own muscular activity via the cardiac circulation (i.e. the venous return to the heart). However, because high temperature favours increased oxygen extraction by skeletal muscle, which consequently lowers venous oxygen, cardiac oxygen supply may become insufficient to meet cardiac oxygen demand. The hypoxic myocardium then cannot maintain cardiac scope and internal oxygen delivery to tissue declines. PMID- 12095865 TI - The force-frequency relationship in fish hearts--a review. PMID- 12095866 TI - Plasticity of excitation-contraction coupling in fish cardiac myocytes. AB - Ultrastructure, molecular composition and electrophysiological properties of cardiac myocytes and functional characteristics of the fish heart suggest that cycling of extracellular Ca(2+) is generally more important than intracellular cycling of Ca(2+) stores of the sarcoplasmic reticulum (SR) in activating contraction of fish cardiac myocytes. This is especially true for the ventricle. However, prominent species-specific differences exist in cardiac excitation contraction coupling and in the relative roles of extracellular and intracellular Ca(2+) sources among the teleostean fish. In fact, in some fish species (tunas, burbot) the SR of atrial myocytes, under certain circumstances, may act as the major source of systolic Ca(2+). These interspecific differences are obviously an outcome of evolutionary adaptation to different habitats and modes of activity in these habitats. There is also substantial intraspecific variation in the SR Ca(2+)-release-to-SL-Ca(2+) influx ratio depending on acute and chronic temperature changes. Consequently excitation-contraction coupling of the fish cardiac myocytes is not a fixed entity, but rather a highly variable and malleable process that enables fish to have an appropriate cardiac scope to exploit a diverse range of environments. PMID- 12095868 TI - Innate visual object recognition in vertebrates: some proposed pathways and mechanisms. AB - Almost all vertebrates are capable of recognizing biologically relevant stimuli at or shortly after birth, and in some phylogenetically ancient species visual object recognition is exclusively innate. Extensive and detailed studies of the anuran visual system have resulted in the determination of the neural structures and pathways involved in innate prey and predator recognition in these species [Behav. Brain Sci. 10 (1987) 337; Comp. Biochem. Physiol. A 128 (2001) 417]. The structures involved include the optic tectum, pretectal nuclei and an area within the mesencephalic tegmentum. Here we investigate the structures and pathways involved in innate stimulus recognition in avian, rodent and primate species. We discuss innate stimulus preferences in maternal imprinting in chicks and argue that these preferences are due to innate visual recognition of conspecifics, entirely mediated by subtelencephalic structures. In rodent species, brainstem structures largely homologous to the components of the anuran subcortical visual system mediate innate visual object recognition. The primary components of the mammalian subcortical visual system are the superior colliculus, nucleus of the optic tract, anterior and posterior pretectal nuclei, nucleus of the posterior commissure, and an area within the mesopontine reticular formation that includes parts of the cuneiform, subcuneiform and pedunculopontine nuclei. We argue that in rodent species the innate sensory recognition systems function throughout ontogeny, acting in parallel with cortical sensory and recognition systems. In primates the structures involved in innate stimulus recognition are essentially the same as those in rodents, but overt innate recognition is only present in very early ontogeny, and after a transition period gives way to learned object recognition mediated by cortical structures. After the transition period, primate subcortical sensory systems still function to provide implicit innate stimulus recognition, and this recognition can still generate orienting, neuroendocrine and emotional responses to biologically relevant stimuli. PMID- 12095867 TI - Effects of temperature and hypercapnia on ventilation and breathing pattern in the lizard Uromastyx aegyptius microlepis. AB - In most reptiles, the ventilatory response to hypercapnia consists of large increases in tidal volume (V(T)), whereas the effects on breathing frequency (f(R)) are more variable. The increased V(T) seems to arise from direct inhibition of pulmonary stretch receptors. Most reptiles also exhibit a transitory increase in ventilation upon removal of CO(2) and this post hypercapnic hyperpnea may consist of changes in both V(T) and f(R). While it is well established that increased body temperature augments the ventilatory response to hypercapnia, the effects of temperature on the post-hypercapnic hyperpnea is less described. In the present study, we characterise the ventilatory response of the agamid lizard Uromastyx aegyptius to hypercapnia and upon the return to air at 25 and 35 degrees C. At both temperatures, hypercapnia caused large increases in V(T) and small reductions in f(R), that were most pronounced at the higher temperature. The post-hypercapnic hyperpnea, which mainly consisted of increased f(R), was numerically larger at 35 compared to 25 degrees C. However, when expressed as a proportion of the levels of ventilation reached during steady-state hypercapnia, the post-hypercapnic hyperpnea was largest at 25 degrees C. Some individuals exhibited buccal pumping where each expiratory thoracic breath was followed by numerous small forced inhalations caused by contractions of the buccal cavity. This breathing pattern was most pronounced during severe hypercapnia and particularly evident during the post hypercapnic hyperpnea. PMID- 12095869 TI - The effect of water deprivation on the expression of atrial natriuretic peptide and its receptors in the spinifex hopping mouse, Notomys alexis. AB - This study investigated the mRNA expression of the atrial natriuretic peptide (ANP) system (peptide and receptors) during water deprivation in the spinifex hopping mouse, Notomys alexis, a native of central and western Australia that is well adapted to survive in arid environments. Initially, ANP, NPR-A and NPR-C cDNAs (partial for receptors) were cloned and sequenced, and were shown to have high homology with those of rat and mouse. Using a semi-quantitative multiplex PCR technique, the expression of cardiac ANP mRNA and renal ANP, NPR-A, and NPR-C mRNA was determined in 7- and 14-day water-deprived hopping mice, in parallel with control mice (access to water). The levels of ANP mRNA expression in the heart remained unchanged, but in the kidney ANP mRNA levels were increased in the 7-day water-deprived mice, and were significantly decreased in the 14-day water deprived mice. NPR-A mRNA levels were significantly higher in 7-day water deprived mice while no change for NPR-A mRNA expression was observed in 14-day water-deprived mice. No variation in NPR-C mRNA levels was observed. This study shows that water deprivation differentially affects the expression of the ANP system, and that renal ANP expression is more important than cardiac ANP in the physiological adjustment to water deprivation. PMID- 12095870 TI - Wetness of the nest environment influences cardiac development in pre- and post natal snapping turtles (Chelydra serpentina). AB - We dissected hearts from near-term embryos and hatchlings of common snapping turtles (Chelydridae: Chelydra serpentina) whose eggs had incubated on wet or dry substrates, and then dried and individually weighed the heart and yolk-free carcass from each animal. Hearts and carcasses of prenatal and neonatal animals grew at different rates, and the patterns of growth by both heart and carcass differed between wet and dry environments. Hearts grew faster, both in actual mass and in mass adjusted for variation in body size, in embryos and hatchlings whose eggs were incubated on dry substrates than in animals whose eggs were held on wet media. This finding is consistent with a hypothesis that embryos incubating in dry settings experience hypovolemia secondary to dehydration and that enlargement of the heart compensates, in part, for the associated increase in viscosity of the blood. Embryonic turtles seemingly exhibit the same plasticity and response that would be expected from other vertebrate ectotherms subjected to the physiological challenges associated with desiccation and an associated reduction in blood volume. PMID- 12095871 TI - Effect of dietary fatty acids on lipoprotein lipase gene expression in the liver and visceral adipose tissue of fed and starved red sea bream Pagrus major. AB - Juvenile red sea bream Pagrus major were fed either a commercial diet (diet 1) or diets supplemented with 10% oleate (diet 2), 5% oleate+5% linoleate (diet 3) or 5% oleate+5% n-3 polyunsaturated fatty acid mixture (diet 4) for 4 weeks. Following the conditioning period, the effects of dietary fatty acids on lipoprotein lipase (LPL) gene expression in the liver and visceral adipose tissue of fed (5 h post-feeding) and starved (48 h post-feeding) fish were investigated by competitive polymerase chain reaction. Fish liver showed substantial LPL mRNA expression that is not found in adult rat liver. When compared with diet 1, diets 2-4 tended to increase the LPL mRNA level in the liver, but tended to decrease it in the visceral adipose tissue under the fed condition. The reciprocal regulation of the liver and visceral adipose LPL mRNA abundance by dietary fatty acids was comparable to that of rat brown and white adipose tissue, respectively. The change in the LPL mRNA level by fatty acids was not completely consistent with the degree of fatty acid unsaturation. Our results indicate that the regulatory effect of dietary fatty acids on LPL gene expression was tissue-specific and related to feeding conditions, but was not solely dependent on the degree of unsaturation of fatty acids. PMID- 12095872 TI - Latex allergy: a health care problem of epidemic proportions. AB - Latex allergy is one of the major health concerns of the decade. The induction of latex allergy commonly occurs after exposure of skin or mucous membrane to natural rubber latex. It is usually a type I, immediate hypersensitivity reaction mediated by IgE which can range from mild skin erythema to anaphylaxis and death. To manage latex allergy appropriately, prompt and correct diagnosis is essential. This depends primarily on clinical history, latex specific IgE RAST (in vitro) and skin prick test (in vivo). It affects people who are frequently exposed to products made of natural rubber latex, such as, health care workers, workers in the rubber industry and those who have had multiple surgical operations. The prevalence in the general population was estimated at 0.7% in a population study in France, but this can be higher in health care workers (up to 17%) and in patients with spina bifida to almost 65%. Prevention strategies should therefore target at risk population, rather than the population as a whole. At present this primarily involves providing non-powdered latex gloves in hospitals and primary health care centres. However, once latex allergy has been established, the situation becomes more complicated. Health care workers either have to be moved to a latex free environment or in extreme circumstances may have to give up their occupation. For patients undergoing operations most hospitals have adopted a latex free protocol. It is important that this is understood and followed by every member of the staff. Those who have had serious adverse reactions should wear Medi-Alert bracelets and in exceptional circumstances may need to keep epinephrine injections on their person. Future strategies focus on the reduction of allergens during latex manufacture, development of suitable non latex gloves and immunotherapy including desensitisation of latex allergic individuals and development of candidate vaccine. PMID- 12095873 TI - Experimental cutaneous Leishmaniasis: a powerful model to study in vivo the mechanisms underlying genetic differences in Th subset differentiation. AB - The murine model of infection with Leishmania major has allowed the demonstration in vivo of the importance CD4+ T cell subsets, distinguishable by the pattern of cytokines they produce, on the outcome of infectious diseases. Genetically determined resistance and susceptibility to infection with this parasite are the result of the development of Th1 and Th2 response, respectively. In this short paper, we present some results obtained in our group pertaining to the analysis of the mechanisms, operational during the early phase of this infection, responsible for the maturation of these functionally distinct CD4+ responses. PMID- 12095874 TI - Segmental lentiginosis with ipsilateral nevus depigmentosus: another example of twin spotting? AB - We report a patient with unilateral lentiginosis and ipsilateral nevus depigmentosus. She had no other cutaneous or extracutaneous abnormalities. The coexistence of segmental lentiginosis and nevus depigmentosus may be explained as a twin-spot phenomenon. PMID- 12095875 TI - Contact sensitivity to hair dyes can be detected by the consumer open test. AB - To avoid allergic reactions to hair dyes, an open test ("skin sensitivity test" or "dab test") is recommended 48 hours before the hair colouring procedure. We have investigated in a multicenter study, under standardized conditions and medical supervision, the validity of this test as a practical method to detect allergy to paraphenylenediamine (PPD)-containing hair dyes in 30 PPD patch test positive and 30 PPD patch test-negative subjects. A defined volume of a marketed hair colouring product containing 1.8% PPD was applied in an open patch test to the retroauricular area. The grading method comprised the dermatologist's clinical evaluation and objective numerical scores. Allergic reactions were elicited in all PPD-positive subjects with a maximal intensity on Day 2; no allergic reactions were elicited in PPD-negative subjects. Increased severity of reactions by the dermatologist's clinical evaluation was correlated strongly with increasing numerical scores. The "skin sensitivity test" can be considered as an effective method to detect type IV hair dye allergy and as such, as an important factor in its secondary prevention. PMID- 12095876 TI - Diphencyprone immunotherapy alters anti-hair follicle antibody status in patients with alopecia areata. AB - Alopecia areata (AA) is a relatively common reversible hair loss disorder usually manifesting as patchy areas of complete hair loss on the scalp and other body parts that can progress to complete loss of all body hair. This condition is now generally assumed to be an autoimmune disease with the hair follicle (HF) as the principal target tissue. AA may be passively transferred by T cells and there is some evidence that serum IgG may also disturb hair cycling. Here, we examine whether the status of anti-HF antibody reactivity is altered during hair regrowth associated with topical immunotherapy using the contact sensitizer diphencyprone. Eleven patients with severe AA of the scalp were treated with diphencyprone on one side of the scalp and serum was obtained from each patient before the start of therapy, after unilateral hair regrowth, during continuing hair regrowth and in some cases after complete and sustained regrowth. The presence and titer of circulating antibodies to HF was assessed by indirect immunofluorescence and immunoblotting analysis. A striking reduction was detected in both the titer and range of HF components/antigens targeted by anti-hair follicle IgG antibodies in those patients that exhibited complete and sustained hair regrowth after DCP treatment. By contrast, unilateral hair regrowth was associated with no change, or even an increase, in anti-HF antibody titer and reactivity. Therefore we can conclude that the down-regulation of antibody reactivity is likely to be a result rather than the cause of hair regrowth induction by topical immunotherapy. As this immunotherapy is associated with a reduction in the titer/pattern of anti-HF antibodies, these may hold the key to the identity of the HF antigen targets in AA. Moreover, the presence/titer of anti-HF antibodies may be a marker of clinical disease activity or opportunity for spontaneous regrowth. PMID- 12095877 TI - The antipsoriatic effect of thiamazole is not accompanied either by significant changes in blood lymphocyte subsets nor by serum concentration of TNF-alpha. AB - It has been reported that the antithyroid thiourethylenes, propylthiouracil and methimazole can bring about significant clinical improvement in patients with psoriasis vulgaris. In this paper, we reconfirmed the clinical usefulness of the antithyroid drug thiamazole, which is similar to methimazole, in Japanese patients with psoriasis vulgaris. We further asked whether the clinical improvement was linked to the changes in some immunological parameters or not. To answer this question, we analyzed peripheral blood lymphocyte subsets and measured serum concentrations of TNF-alpha and IL-1beta in 13 patients with psoriasis vulgaris before and during thiamazole administration. The mean percentage of CD4+ HLA-DR+ lymphocytes in the patients was constantly elevated before and 12 weeks after the administration. The patients treated with thiamazole showed a significant increase in the ratio of CD4+/CD8+ at the 4th week, though it declined at the 8th and 12th week. Other lymphocyte subsets showed no significant change during the treatment. The concentrations of serum TNF-alpha exhibited no significant change during thiamazole administration. Serum concentration of IL-1beta was under the detection level. These results indicate that thiamazole may improve the psoriatic lesion via other immune mechanisms, or the effects can not be reflected by the peripheral lymphocyte subsets and cytokine which we investigated. PMID- 12095878 TI - Effect of activated human mast cells and mast cell-derived mediators on proliferation, type I collagen production and glycosaminoglycans synthesis by human dermal fibroblasts. AB - Although an increased number of mast cells in fibrotic tissues such as scleroderma, keloid or healing wound has been highlighted, it is still unclear whether or not mast cells are fibrogenic. The aim of the present study is to determine whether functionally active human mast cells can provide human dermal fibroblasts directly with fibrogenic properties. In order to examine the effects of IgE-mediated mast cell activation on fibroblast proliferation and synthesis of type I collagen, we utilized an in vitro defined system in which cultured human mast cells were co-cultured with human dermal fibroblasts. We also employed a three-dimensional fibroblast culture system using supplementation of L-ascorbic acid as an assay system to investigate the effects of mast cell-derived mediators on synthesis of glycosaminoglycans by human fibroblast. Fibroblast proliferation was actively stimulated with IgE-activated mast cells. However, this stimulatory effect was canceled in co-cultures with a higher number of IgE-activated mast cells. In the presence of a higher number of activated mast cells, the fibroblast cell layer was destroyed, in contrast to an intact cell layer in the presence of same number of the mast cells without activation. Type I collagen synthesis was unchanged in fibroblasts co-cultured with mast cells. The total amount of main disaccharide units, particularly DELTADi-HA, was increased when fibroblasts were exposed to histamine. Thus, we conclude that other factors, in addition to mast cells, are important in the development of human tissue fibrosis or sclerosis. PMID- 12095879 TI - Treatment of non-genital warts with topical imiquimod 5% cream. AB - Common warts (verrucae vulgaris) are associated with human papillomavirus infection and are routinely treated by ablative procedures such as cryotherapy, electrodessiccation and salicylic acid. We report 10 cases of recurrent warts treated with a potential new topical therapy, imiquimod 5% cream. Nine of the 10 patients were successfully treated with imiquimod 5% cream applied, under occlusion, once daily for 4 weeks. No recurrences were reported during 3 months of follow up. PMID- 12095880 TI - Randomised, controlled trial of the efficacy and safety of adapalene gel 0.1% and tretinoin cream 0.05% in patients with acne vulgaris. AB - BACKGROUND: Previous clinical trials have shown that adapalene gel produces less irritation than tretinoin gels and tretinoin 0.025% cream. Short term results have shown that adapalene is less irritating than tretinoin gels and creams. This study is the first to compare the 0.1% formulation of adapalene gel with the 0.05% strength of tretinoin cream in a formal clinical trial. OBJECTIVE: To investigate the efficacy and tolerability of adapalene gel 0.1% compared with tretinoin cream 0.05% in patients with mild-to-moderate acne vulgaris. METHODS: Ten-week, multicentre, randomised, investigator-masked, active-controlled, parallel group study in 409 patients with acne vulgaris. RESULTS: Adapalene gel 0.1% demonstrated equivalent efficacy in reduction of acne lesion counts and global improvement of acne severity over 10 weeks' treatment and was significantly better tolerated than tretinoin cream 0.05% in terms of erythema, dryness, desquamation and stinging/burning. CONCLUSION: Adapalene gel 0.1% showed equivalent efficacy and was significantly better tolerated than tretinoin cream 0.05% in patients with mild-to-moderate acne vulgaris. PMID- 12095881 TI - Paraneoplastic pemphigus associated with Castleman's disease and asymptomatic bronchiolitis obliterans. AB - We describe a 19-year-old male patient who presented with recalcitrant erosions limited to the orolabial and genital mucosa for 18 months. The clinicopathologic diagnosis of paraneoplastic pemphigus was confirmed by indirect immunofluorescence staining of murine bladder epithelium, and the presence of IgG autoantibodies against envoplakin, periplakin, and 170 kDa protein. Enzyme-linked immunosorbent assay (ELISA) with desmoglein 3 (Dsg3) and desmoglein 1 (Dsg1) recombinant proteins showed that the patient's sera were positive against Dsg3. Castleman's tumour was found in the pelvic cavity and resected completely. The orogenital erosions disappeared slowly after the resection of Castleman's tumour with the concomitant decrease in intercellular antibody titre and index values of Dsg ELISA. Although the patient was unaware of dyspnea on exertion, a notable air flow obstruction persisted over 17 months. The expiratory images of high resolution computed tomography showed air trapping, indicating the presence of asymptomatic but gradually progressive bronchiolitis obliterans. PMID- 12095882 TI - Livedoid vasculopathy and skin ulcers in patients with inherited thrombophilia. PMID- 12095883 TI - Graft-versus-host disease-like eruption preceding a nodal CD30+ lymphoma. PMID- 12095884 TI - Tinea capitis due to Microsporum gypseum. PMID- 12095885 TI - Piezogenic pedal and palmar papulosis. PMID- 12095886 TI - Erythema multiforme after resolution of herpes zoster by acyclovir. PMID- 12095887 TI - Pyogenic granuloma within port-wine stains: an alarming clinical presentation. PMID- 12095889 TI - Hypersensitivity to mosquito bites with natural-killer cell lymphocytosis: the possible implication of Epstein-Barr virus reactivation. AB - Hypersensitivity to mosquito bites (HMB) has been recognized as one of the wide variety of Epstein-Barr virus (EBV)-associated lymphoproliferative disorders, some of which overlap with chronic active EBV infection and hydroa vacciniforme. It frequently predisposes to hemophagocytic syndrome or lymphoid malignancy with a clonal EBV replication. Here we report an 8-year-old Japanese boy who presented with a 3-year history of HMB. Every summer, he had repeated severe local skin reactions with high-grade fever, lymphadenopathy, and mild liver dysfunction after mosquito bites. Histologically, lymphoid cells overexpressing EBV-encoded small nuclear RNAs (EBER), a marker of latent EBV infection, and eosinophils infiltrated diffusely into the lesional skin. His blood tests showed a marked increase in the CD11b+ 16+ 56+ natural-killer cell population (~ 50% of the peripheral lymphocytes), but neither a clonal proliferation of EBV nor abnormal antibody titers to EBV. Quantitative PCR using a cloned EBV-determined nuclear antigen (EBNA)-1 vector revealed that a high copy number of the EBV genome is integrated into his peripheral blood cells even during the mosquito-free winter. Based on this evidence, he was diagnosed as being in a smoldering stage of HMB, in which EBV-associated NK cell lymphoproliferation might participate in its pathogenesis. PMID- 12095888 TI - Parotideal lymph node metastasis in squamous cell carcinoma of the skin. AB - BACKGROUND: Metastatic involvement of the parotideal lymph nodes from cutaneous squamous cell carcinoma is rare in occurrence, but has a high prognostic value. The aim of the present study was to define a patient group with a high risk for development of regional metastasis and to determine the follow-up course and therapy of metastasis in these patients. MATERIAL AND METHODS: Nineteen patients treated with malignancies of the parotid gland over a time period of four years were analyzed prospectively. RESULTS: In 6 out of 19 patients the parotideal tumor proved to be a lymph node metastasis of previously treated poorly differentiated squamous cell carcinoma of the skin. The diameter of the primary tumor was at least 1.5 cm in 5 out of 6 cases. The time interval between detection of metastatic involvement of the parotid gland and diagnosis of a preexisting skin cancer was approximately 7 months. Metastastic infiltration of cervical lymph nodes could be shown in 4 patients. In one patient pulmonary metastases were detected. CONCLUSION: On the basis of data from the literature and the results presented here, patients who are at high risk for regional metastasis were defined. Clinical examination of the parotid gland and cervical lymph nodes should be performed frequently in these patients at least for 18 months after primary tumor diagnosis. Parotideal lymph node metastases of a squamous cell carcinoma of the head skin should have similar treatment to primary squamous cell carcinoma of the parotid gland provided that a curative option exists. PMID- 12095890 TI - Drug eruption and liver injury caused by terfenadine and oxatomide. PMID- 12095891 TI - Use of 0.1% tacrolimus ointment in patients with various forms of lupus erythematosus. PMID- 12095893 TI - Anatomy, histology and immunohistochemistry of normal human skin. AB - The skin is the largest organ of the body, accounting for about 15% of the total body weight in adult humans. It exerts multiple vital protective functions against environmental aggressions, rendered possible thanks to an elaborate structure, associating various tissues of ectodermal and mesodermal origin, arranged in three layers, including (from top to bottom) the epidermis (and its appendages), the dermis and the hypodermis. This article reviews the main data concerning the anatomy, histology and immunohistochemistry of normal human skin. PMID- 12095894 TI - Theme issue: suicidal behaviors in older adults. PMID- 12095895 TI - Respectful decisions at the end of life. PMID- 12095896 TI - The illusion of patient choice in end-of-life decisions. AB - After passage of the Patient Self-Determination Act and the Cruzan decision by the Supreme Court, honoring individual patient choice has become the primary means by which we have sought to improve the quality of life of the dying patient. However, the decision-making capacity of the dying patient is usually compromised, and advance directives have not consistently improved the dying process. We respect patient autonomy in order to respect the patient as a person; patient autonomy should be respected to the degree that it is intact. When autonomy is significantly diminished, as it usually is in dying patients, respecting autonomy reconstructed from documents or proxies may not be the best way to respect the dying person. We rather need to seek social consensus about when patients are dying, the nature of a "good death," and when it is preferable to a longer life. PMID- 12095897 TI - Methodological aspects of comparative research in the epidemiology of Alzheimer disease. AB - In recent years, major advances have been made in the investigation of the incidence and prevalence of Alzheimer disease in diverse social and cultural settings. Examples of issues being addressed are the methods used to estimate incidence and prevalence, the procedures for data collection and its harmonization, the definitions of numerator (cases) and denominator (population at risk), and the modeling of cognitive decline. This type of research will eventually lead to a better understanding of how protective and modifiable risk factors interact with the genotype to produce this disease. PMID- 12095898 TI - Religion, senescence, and mental health: the end of life is not the end of hope. AB - The authors review epidemiological and survey research relevant to the relationships between religiousness/spirituality and mental health in people at the end of life, with the end of helping psychiatrists, psychologists, and other mental health professionals dealing with older Americans. They give special attention to well-being, religious coping, cognitive dysfunction, anxiety, depression, and suicide, and consider the extent to which hope is a mediator of the purported salutary effects of religiousness. Studies were selected from the comprehensive and systematic review of 20th-century scientific literature concerning religion and health. Authors also review current studies relevant to religion and end-of-life issues. Religious persons reported generally higher levels of well-being. The review also found fairly consistent inverse associations of religiousness with rates of depression and suicide. There was some negative association between religious participation and cognitive dysfunction, but the association with anxiety was inconsistent, with some studies showing a correlation between higher levels of religion and anxiety. Religion's effects on mental health are generally protective in direction but modest in strength. PMID- 12095899 TI - Risk factors for late-life suicide: a prospective, community-based study. AB - Despite the fact that people age 65 and older have the highest rates of suicide of any age-group, late-life suicide has a low prevalence, making it difficult to conduct prospective studies. The authors examined risk factors for late-life suicide on the basis of general information collected directly from older subjects participating in a community-based prospective study of aging, the Established Populations for Epidemiologic Studies of the Elderly. Demographic variables, presence of a relative or friend to confide in, alcohol use, and sleep quality were assessed at baseline interview. Baseline and follow-up data were used to determine physical, cognitive, and affective functioning, as well as medical status. Of 14,456 people, 21 committed suicide over the 10-year observation period. Depressive symptoms, perceived health status, sleep quality, and absence of a relative or friend to confide in predicted late-life suicide. Suicide victims did not have greater alcohol use and did not report more medical illness or physical impairment. This study provided additional information about the context of late-life depression that also contributes to suicidal behavior: poor perceived health, poor sleep quality, and limited presence of a relative or friend to confide in. PMID- 12095900 TI - Access to firearms and risk for suicide in middle-aged and older adults. AB - Elderly white men are at the highest risk for suicide. Firearms are the most common method of suicide used by both men and women in later life, and a greater proportion of older than younger suicide victims use a gun. This psychological autopsy study aimed to test hypotheses concerning the risk for suicide associated with access to and storage of firearms. Subjects included 86 suicide victims age 50 years of age and over and 86 community control subjects individually matched on age, sex, race, and county of residence. Presence of a firearm in the home was associated with increased risk for suicide, even after controlling for psychiatric illness. Elevated risk was accounted for by access to handguns rather than long guns and was more pronounced in men than women. Among subjects who kept a gun in the home, storing the weapon loaded and unlocked were independent predictors of suicide. Findings support the potential benefit for suicide prevention of restricting access to handguns. Education programs for older persons, their families, and healthcare providers concerning the risks of having a gun in the home and reinforcement of rules for safe storage may contribute to reducing the rate of suicide in older people. PMID- 12095902 TI - Cognitive functioning and geriatric suicide ideation: testing a mediational model. AB - The authors evaluated a structural model of the relationship between cognitive functioning and geriatric suicide ideation, mediated by Depression/Hopelessness, a latent construct characterized by a combination of the two. A heterogeneous sample of 90 elderly participants completed the Geriatric Suicide Ideation Scale, a new, multidimensional self-report measure of suicide ideation in seniors, and measures of cognitive functioning, depression, and both global and social forms of hopelessness. The results demonstrated significant associations between suicide ideation and the various predictor variables. The results of structural equation modeling supported the proposed mediational model, indicating that the Depression/Hopelessness construct mediates the relationship between cognitive functioning and suicide ideation among older persons. These findings have implications for the conceptualization and treatment of potentially suicidal elderly persons. PMID- 12095901 TI - Suicidal and death ideation in older primary care patients with depression, anxiety, and at-risk alcohol use. AB - The authors identified correlates of active suicidal ideation and passive death ideation in older primary care patients with depression, anxiety, and at-risk alcohol use. Participants included 2,240 older primary care patients (age 65+), who were identified in three mutually exclusive groups on the basis of responses to the Paykel suicide questions: No Ideation, Death Ideation, and Suicidal Ideation. Chi-square, ANOVA, and polytomous logistic regression analyses were used to identify characteristics associated with suicidal ideation. The highest amount of suicidal ideation was associated with co-occurring major depression and anxiety disorder (18%), and the lowest proportion occurred in at-risk alcohol use (3%). Asians have the highest (57%) and African Americans have the lowest (27%) proportion of suicidal or death ideation. Fewer social supports and more severe symptoms were associated with greater overall ideation. Death ideation was associated with the greatest medical comorbidity and highest service utilization. Contrary to previous reports, authors failed to find that active suicidal ideation was associated with increased contacts with healthcare providers. Accordingly, targeted assessment and preventive services should be emphasized for geriatric outpatients with co-occurring depression and anxiety, social isolation, younger age, and Asian or Caucasian race. PMID- 12095903 TI - Suicidality in older African Americans: findings from the EPOCH study. AB - The authors examined the current frequency of suicidality and associated characteristics in a sample of 835 African-American older adult residents of six urban public housing developments who consented to participate in an intervention trial of mobile outreach. The frequency of passive and active suicidal ideation was 2.5% and 1.4%, respectively. Characteristics of individuals with both active and passive suicidality included elevated anxiety, social dysfunction, somatic symptoms, low social support, lack of a confidant, and low religiosity. Characteristics of those with passive, but not active, ideation also included older age, lower levels of education, elevated depressive symptoms, poorer cognitive functioning, and having recently discussed emotional problems with a healthcare provider. The characteristics of those reporting active, but not passive, ideation included having a history of mental health treatment and reporting no instrumental support. Multivariate analyses indicated that depression and religiosity were uniquely associated with passive suicidal ideation, and life satisfaction and religiosity were uniquely associated with active suicidal ideation. The authors discuss implications of these findings and offer suggestions for research and clinical practice. PMID- 12095904 TI - Psychiatric disorders among bereaved persons: the role of perceived circumstances of death and preparedness for death. AB - The authors evaluated the association between bereaved persons' perceptions of the death (e.g., extent of suffering) and preparedness for the death and psychiatric disorders. The sample was 122 persons who received a baseline interview at approximately 4 months post-loss and a follow-up interview 5 months later, at approximately 9 months post-loss. In addition to bivariate associations between perceptions of circumstances of death and psychiatric outcomes, authors used multivariate logistic-regression models to determine the extent to which perceptions of the death were associated with the likelihood of psychiatric disorders in survivors at baseline and follow-up. Perception of the death as more violent was associated with major depressive disorder at baseline. Perception of lack of preparedness for the death was associated with complicated grief at baseline and at follow-up. These preliminary results suggest that perceptions of the death and feelings of lack of preparedness for it may be indicators of persons at risk of developing psychiatric morbidity secondary to bereavement. Further studies are needed to confirm these findings. PMID- 12095905 TI - Predictors of two-year mortality in a prospective "UPBEAT" study of elderly veterans with comorbid medical and psychiatric symptoms. AB - Medical inpatients of nine VA medical centers (N=2,657) were screened for symptoms of depression, anxiety, and alcohol abuse and followed for 24 months. Survivors were compared with deceased subjects on the severity of symptoms of depression, anxiety, alcohol abuse, and self-rated health. Mortality was predicted by the length of hospitalization, as well as poor self-rated health at baseline. The severity of depressive symptoms and poor self-rated health measured at the time closest to the time of death also predicted mortality. Lack of improvement in symptoms of depression and anxiety at 6 months was associated with higher rates of mortality. PMID- 12095906 TI - Treatment of homebound mentally ill elderly patients: the multidisciplinary psychiatric mobile team. AB - Authors evaluated the general feasibility of using a multidisciplinary psychiatric mobile team model for homebound elderly patients. Information was collected with a semistructured interview on 235 homebound elderly patient referrals over a 2.5-year period. Ninety-three subjects were accepted into the program; 12.9% of those accepted required psychiatric hospitalization during the follow-up. Maintaining individuals in the community was achieved with intense case management and significant increases in home-care hours. The multidisciplinary team approach appeared to lessen psychiatric disability in homebound mentally ill elderly patients. PMID- 12095907 TI - Valproate-induced hyperammonemia as a cause of altered mental status. AB - The authors describe valproate-induced hyperammonemia and mental status changes in an 88-year-old man, the first known reported case in an elderly patient. They discuss this underrecognized complication of valproate use and the implications for treating elderly patients, in whom valproate use is increasing. PMID- 12095909 TI - Suicide and Alzheimer disease. PMID- 12095908 TI - Alzheimer disease, serotonin systems, and tryptophan depletion. PMID- 12095910 TI - Sertraline-related night sweats. PMID- 12095912 TI - The DNA mismatch-repair MLH3 protein interacts with MSH4 in meiotic cells, supporting a role for this MutL homolog in mammalian meiotic recombination. AB - The mismatch-repair (MMR) system plays a central role in maintaining genetic stability and requires evolutionarily conserved protein factors, including MutS and MutL homologs. Since the discovery of a link between the malfunction of post replicative MMR and human cancers, a number of works have focused on the function of MutS and MutL homologs in the correction of replication errors. However, several MutS-like and MutL-like proteins also participate in meiotic recombination. The MutL homolog MLH3 has been recently identified in mammals. Several pieces of evidence support a role for this protein in post-replicative MMR. To investigate whether MLH3 also acts during meiotic recombination, we analyzed its expression in mammalian germ cells. The MLH3 gene is expressed in mouse meiotic cells and in human testis, and, as revealed by immunoprecipitation assays, the MLH3 protein is found in mouse spermatocytes. We further demonstrate that the meiosis-specific MSH4 protein, known to participate to meiotic recombination, is co-immunoprecipitated with MLH3 from mouse meiotic cell extracts. In addition, the two MLH3 protein isoforms potentially expressed in human testis (hMLH3 and hMLH3 Delta 7) interact in vitro with the hMSH4 protein. These interaction data suggest that MLH3 is associated with MSH4 in mammalian meiotic cells, and strongly support the possibility that MLH3 plays a role in mammalian meiotic recombination. PMID- 12095911 TI - PTEN blocks insulin-mediated ETS-2 phosphorylation through MAP kinase, independently of the phosphoinositide 3-kinase pathway. AB - The tumor suppressor PTEN possesses lipid and protein phosphatase activities. It has been well established that the lipid phosphatase activity is essential for its tumor-suppressive function via the phosphoinositide 3-kinase (PI3K) and Akt pathways. The precise role of the protein phosphatase activity is still unclear. In the current study, we demonstrate that overexpression of wild-type PTEN in the MCF-7 breast cancer line results in phosphatase activity-dependent decreases in the phosphorylation of ETS-2, which is a transcription factor whose DNA-binding ability is controlled by phosphorylation. Exposure of MCF-7 cells to insulin, insulin-like growth factor 1 (IGF-1) or epidermal growth factor (EGF) can lead to the phosphorylation of ETS-2, Akt and ERK1/2. The MEK inhibitor PD590089 abrogates insulin-stimulated phosphorylation of ETS-2. In contrast, the PI3K inhibitor LY492002 has no effect on insulin-stimulated phosphorylation of ETS-2, despite the fact that it diminishes insulin-stimulated phosphorylation of Akt. Interestingly, overexpression of PTEN in MCF-7 leads to blockade of insulin stimulated, but not EGF-stimulated, phosphorylation of ERK, accompanied by dramatic decreases in ETS-2 phosphorylation. We further show that the relationship of PTEN and ETS-2 has functional significance by demonstrating that PTEN abrogates activation of the uPA Ras-responsive enhancer, a target of ETS-2 action, in a phosphatase-dependent manner, irrespective of the presence or absence of insulin. Our observations, therefore, suggest that PTEN blocks insulin stimulated ETS-2 phosphorylation through inhibition of the ERK members of the MAP kinase family independently of PI3K, and that the PTEN effect on the phosphorylation status of ETS-2 may be mediated through PTEN's protein phosphatase activity. PMID- 12095913 TI - Allele-specific expression analysis by RNA-FISH demonstrates preferential maternal expression of UBE3A and imprint maintenance within 15q11- q13 duplications. AB - 15q11- q13 contains many imprinted genes, and undergoes duplicon-mediated rearrangements, including deletions, duplications and triplications, and generation of marker chromosomes. Abnormal phenotypes, including language delays and autism spectrum disorders, are primarily observed with maternal 15q11- q13 duplication. To determine possible epigenetic effects on expression within duplicated 15q11- q13 regions, we utilized RNA-FISH to directly observe gene expression. RNA-FISH, unlike RT-PCR, is polymorphism-independent, and it also detects relative levels of expression at each allele. Unamplified, gene-specific RNA signals were detected using cDNA probes. Subsequent DNA-FISH confirmed RNA signals and assigned parental origin by colocalization of genomic probes. SNRPN and NDN expression was detected primarily from paternal alleles. Control Dystrobrevin transcripts were detected equally from both alleles; however, maternal-UBE3A signals were consistently larger than paternal signals in normal fibroblasts and in neural-precursor cells. Larger UBE3A signals were also observed on one or both maternal alleles in a cell line carrying a maternal interstitial duplication, on both alleles of a maternally derived marker(15) chromosome, and occasionally on a paternal allele in a cell line carrying a paternal interstitial duplication. Expression of NDNL2, just distal to the duplicated region, was not markedly altered but paralleled changes in UBE3A expression. Excess total maternal-UBE3A RNA was confirmed by Northern blot analysis of cell lines carrying 15q11- q13 duplications or triplications. These results demonstrate that: (1) UBE3A is imprinted in fibroblasts, lymphoblasts and neural-precursor cells; (2) allelic imprint status is maintained in the majority of cells upon duplication both in cis and in trans; and (3) alleles on specific types of duplications may exhibit an increase in expression levels/loss of expression constraints. PMID- 12095914 TI - Stable micro-dystrophin gene transfer using an integrating adeno-retroviral hybrid vector ameliorates the dystrophic pathology in mdx mouse muscle. AB - The ability to transfer the dystrophin gene stably to the skeletal muscle of DMD patients is a major confounding issue in establishing an effective gene therapy for this disease. To overcome this problem, we have examined the ability of muscle fibres from mdx mice to act as in situ factories of retroviral vector production. Tibialis anterior (TA) muscles from 4-week-old mdx mice were injected with an adenoviral vector expressing LacZ within a retroviral expression cassette (AdLZIN). Retroviral vector production was induced by the inclusion of two additional adenoviral vectors expressing retroviral gag-pol (AdGagPol) and 10A1 env genes (Ad10A1). Upon introduction of infected muscles into cell culture, colonies of beta-galactosidase-expressing myotubes formed only in cultures where the muscle was injected with AdLZIN, AdGagPol and Ad10A1, but not from muscle injected with AdLZIN only. Muscles from mdx/nude mice producing retroviral vector displayed a 4.6-fold increase in beta-galactosidase-positive myofibres after 1 month, compared with contralateral muscle in the same animal injected with AdLZIN and AdGagPol only. By constructing a hybrid adeno-retroviral vector expressing a truncated micro-dystrophin construct (AdmicroDyIN), we were able to partially correct the mdx dystrophic phenotype. AdmicroDyIN-mediated expression of micro dystrophin in mdx TA muscle restored the formation of the dystrophin-associated glycoprotein complex and significantly reduced the level of muscle degeneration over uninjected controls. By stimulating in situ production of retroviral vector expressing micro-dystrophin, we achieved 92%+/-6% transduction of myofibres in the TA muscle by 4 weeks. Strikingly, by 3 months post injection, micro dystrophin was still expressed to high levels in nearly all the myofibres of the TA muscle. By comparison, there was a pronounced drop in the levels of micro dystrophin expressed by muscles injected with AdmicroDyIN only. Finally, using a novel PCR approach, we detected reverse-transcribed, integrated proviral sequences in TA muscle genomic DNA by 4 weeks post injection, the levels of which were found to increase after 3 months. PMID- 12095916 TI - Identification and characterization of an imprinted antisense RNA (MESTIT1) in the human MEST locus on chromosome 7q32. AB - Imprinted gene(s) on human chromosome 7 are thought to be involved in Russell Silver syndrome (RSS), based on the fact that approximately 10% of patients have maternal uniparental disomy of chromosome 7. However, involvement of the known imprinted genes (GRB10 at 7p12, PEG10 at 7q21.3 and MEST at 7q32) in RSS has yet to be established. To screen for new imprinted genes, we are initially using somatic cell hybrids containing a paternal or maternal human chromosome 7. Transcripts located between D7S530 and D7S649 (a 1.5 Mb interval encompassing MEST ) were subjected to RT-PCR analysis using somatic cell hybrids. One transcript named MESTIT1 (for MEST intronic transcript 1) reproducibly showed paternal-specific expression. Upon further analysis, we found MESTIT1 to be (1) paternally (and not maternally) expressed in all fetal tissues and fibroblasts examined, (2) to be located in an intron of one of the two isoforms of MEST but transcribed in the opposite direction, (3) to be composed of at least two exons without any significant open reading frame, and (4) to exist as a 4.2 kb transcript in many fetal and adult tissues. We could also identify two isoforms of the mouse Mest gene as observed in humans, but it is still unknown if a murine ortholog of MESTIT1 exists. We also examined the imprinting status of MEST isoforms as a first step in assessing whether MESTIT1 might influence the allelic expression pattern of the sense transcript. MEST isoform 1 was determined to be exclusively expressed from the paternal allele in all fetal tissues and cell lines examined, whereas MEST isoform 2 was only preferentially expressed from the paternal allele in a tissue/cell-type-specific manner. Our results suggest that MESTIT1 is a paternally expressed non-coding RNA that may be involved in the regulation of MEST expression during development. MESTIT1 (also known as PEG1-AS) is now the third independent transcript (with MEST and COPG2IT1) identified at human chromosome 7q32 demonstrating paternal chromosome-specific expression. PMID- 12095915 TI - Pioglitazone improves the phenotype and molecular defects of a targeted Pkd1 mutant. AB - Mutations of either PKD1 or PKD2 are associated with autosomal dominant polycystic kidney disease (ADPKD). The molecular function of the gene product of PKD1, polycystin-1, in vitro has been elucidated recently, but the molecular pathological consequences of the loss of polycystin-1 in vivo have remained unclear. We have generated a mouse with a targeted deletion of exons 2-6 of Pkd1 to study the molecular defects in Pkd1 mutants. Homozygote embryos (Pkd1(-/-)) developed hydrops, cardiac conotruncal defects and renal cystogenesis. Total protein levels of beta-catenin in heart and kidney and c-MYC in heart were decreased in Pkd1(-/-) embryos. In the kidneys of Pkd1(-/-), the expression of E cadherin and PECAM in basolateral membranes of renal tubules was attenuated, and tyrosine phosphorylation of epidermal growth factor receptor and Gab1 were constitutively enhanced when cystogenesis started on embryonic day (E) 15.5-16.5. Maternally administered pioglitazone, a thiazolidinedione compound, resolved these molecular defects of Pkd1(-/-). Treatment with pioglitazone improved survival of Pkd1(-/-) embryos and ameliorated the cardiac defects and the degree of renal cystogenesis. Long-term treatment with pioglitazone improved the endothelial function of adult Pkd1(+/-). These data indicated that molecular defects observed in Pkd1(-/-) embryos contributed to the pathogenesis of ADPKD and that thiazolidinediones had a compensatory effect on the pathway affected by the loss of polycystin-1. Pathways activated by thiazolidinediones may provide new therapeutic targets in ADPKD. PMID- 12095917 TI - A common protein interaction domain links two recently identified epilepsy genes. AB - Until recently, all genes found to be mutated in hereditary idiopathic epilepsies encoded subunits of ion channels, leading to the view of this class of diseases as channelopathies. Two apparent exceptions to this rule are the MASS1 gene, which is mutated in the Frings mouse model of audiogenic epilepsy, and the LGI1 gene, which is mutated in autosomal dominant partial epilepsy with auditory features (ADPEAF). Careful sequence analysis of the two protein products encoded by those genes shows a common feature: both sequences harbour a novel homology domain consisting of a 7-fold repeated 44-residue motif. The architecture and structural features of this new domain make it a likely member of the growing class of protein interaction domains with a seven-bladed beta-propeller fold. In the MASS1 gene product, which has recently been shown to be a fragment of the very large G-protein-coupled receptor VLGR1, this EAR domain (for epilepsy associated repeat) is part of the ligand-binding ectodomain. LGI1, as well as a number of newly identified LGI1 relatives, is predicted to be a secreted protein, and consists of an N-terminal leucine-rich repeat region and a C-terminal EAR region. The known portion of the human genome encodes six EAR proteins, some of which map to chromosome regions associated with seizure disorders. The EAR domain is likely to play an important role in the pathogenesis of epilepsy, either by binding to an unknown anti-epileptic ligand, or more likely by interfering with axon guidance or synaptogenesis. PMID- 12095918 TI - EDA targets revealed by skin gene expression profiles of wild-type, Tabby and Tabby EDA-A1 transgenic mice. AB - Mutations in the EDA gene cause anhidrotic ectodermal dysplasia (EDA), with lesions in skin appendage formation. To begin to analyze EDA pathways, we have used expression profiling on 15,000-gene mouse cDNA microarrays, comparing adult mouse skin from wild-type, EDA-defective (Tabby) mice, and Tabby mice supplemented with the EDA-A1 isoform, which is sufficient to rescue multiple Tabby phenotypes. Given the sensitivity of the current microarray system, 8500 genes (60%) were estimated to be expressed, including transcription factors and growth-regulatory genes that had not previously been identified in skin; but only 24 (0.16%), one-third of them novel, showed significant differences between wild type and Tabby. An additional eight genes not included in the 15,000 gene set were shown to have expression differences by real-time RT-PCR. Sixteen of 32 affected genes were restored significantly toward wild-type levels in EDA-A1 transgenic Tabby mice. Significant up-regulation in Tabby skin was observed for several dermal matrix genes, including Col1a1, Col1a2, Col3a1 and SPARC: In contrast, down-regulation occurred for the NEMO/NF-kappa B pathway, already implicated in skin appendage formation, and even more markedly for a second pathway, JNK/c-jun/c-fos and their target genes, that has not previously been clearly associated with skin development. These data are consistent with the regulation of the NF-kappa B pathway by EDA, and support its involvement in the regulation of the JNK pathway as well. PMID- 12095919 TI - Osteopenia and male-specific sudden cardiac death in mice lacking a zinc transporter gene, Znt5. AB - We isolated a mammalian gene whose expression transiently increased in response to intimal denudation of rabbit aorta. It was identical to a gene encoding a zinc transporter, ZNT5, reported very recently by others. Mice deficient for this gene showed poor growth and a decrease in bone density due to impairment of osteoblast maturation to osteocyte. More than 60% of male null mice died suddenly because of the bradyarrhythmias. Analysis of gene-expression profiles in murine hearts by means of an oligonucleotide microarray disclosed that a subset of genes encoding immediate-early response factors (IEGs) and heat shock proteins (HSPs) were down regulated in Znt5-null mice. These results indicate that Znt5 protein plays an important role in maturation of osteoblasts and in maintenance of the cells involved in the cardiac conduction system, partly owing to dysregulated expression of IEGs and HSPs. PMID- 12095920 TI - SMN, the spinal muscular atrophy protein, forms a pre-import snRNP complex with snurportin1 and importin beta. AB - The survival of motor neuron (SMN) protein is mutated in patients with spinal muscular atrophy (SMA). SMN is part of a multiprotein complex required for biogenesis of the Sm class of small nuclear ribonucleoproteins (snRNPs). Following assembly of the Sm core domain, snRNPs are transported to the nucleus via importin beta. Sm snRNPs contain a nuclear localization signal (NLS) consisting of a 2,2,7-trimethylguanosine (TMG) cap and the Sm core. Snurportin1 (SPN) is the adaptor protein that recognizes both the TMG cap and importin beta. Here, we report that a mutant SPN construct lacking the importin beta binding domain (IBB), but containing an intact TMG cap-binding domain, localizes primarily to the nucleus, whereas full-length SPN localizes to the cytoplasm. The nuclear localization of the mutant SPN was not a result of passive diffusion through the nuclear pores. Importantly, we found that SPN interacts with SMN, Gemin3, Sm snRNPs and importin beta. In the presence of ribonucleases, the interactions with SMN and Sm proteins were abolished, indicating that snRNAs mediate this interplay. Cell fractionation studies showed that SPN binds preferentially to cytoplasmic SMN complexes. Notably, we found that SMN directly interacts with importin beta in a GST-pulldown assay, suggesting that the SMN complex might represent the Sm core NLS receptor predicted by previous studies. Therefore, we conclude that, following Sm protein assembly, the SMN complex persists until the final stages of cytoplasmic snRNP maturation and may provide somatic cell RNPs with an alternative NLS. PMID- 12095922 TI - The Amazon Project: helping the patient to become an instrument of her own recovery. AB - The Amazon Project combines science, myth, and theater to explore problems linked to cancer and its surgical management in women from a social, political, and cultural point of view. This Project is concerned with change in the hope that one day it will be possible to discuss, along with the Science of Medicine, the Science of Patients as well. PMID- 12095923 TI - Gene therapy of hepatocellular carcinoma and gastrointestinal tumors. AB - Primary liver cancer and liver metastases from gastrointestinal tumors lack effective therapy. Gene therapy is a promising therapeutic approach and is based on the introduction of genetic material into cells to generate a curative biological effect. Adenoviral vectors can very efficiently transduce a wide variety of malignant epithelial cells both in vitro and in vivo. A variety of gene therapy-based anticancer strategies have been effective in animal tumor models, including replacement of tumor suppressor genes, selective activation of prodrugs, genetic immunotherapy, and antiangiogenic actions. Enzymes used for genetic activation include viral thymidine kinase (tk), which may activate nucleoside analogs such as ganciclovir. We and others have demonstrated the efficacy of the tk/ganciclovir system in the treatment of hepatocellular carcinoma and metastatic colorectal cancer in experimental models. Also, this strategy can be safely applied to patients with liver tumors. Interleukin-12 (IL 12) is among the most potent cytokines in stimulating antitumor immunity. In models of primary and metastatic liver cancer we showed that intratumoral administration of recombinant adenovirus encoding IL-12 activates natural killer cells, induces specific antitumor immunity, and displays a powerful antiangiogenic effect, resulting in tumor regression. There is a synergistic effect with the gene transfer of the chemokine IP-10. Also, intratumoral injection of either dendritic cells transfected ex vivo with recombinant adenovirus encoding IL-12 (Ad.IL-12) or an adenovirus coding for the CD40 ligand have shown an intense antitumor effect against experimental colorectal cancer. In summary, a variety of gene therapy strategies have been effective against animal models of gastrointestinal tumors. Clinical trials should determine whether human patients can be treated safely and effectively by such strategies. PMID- 12095924 TI - Epidemiology, risk factors, and natural history of hepatocellular carcinoma. AB - The incidence of hepatocellular carcinoma is increasing in many countries. The estimated number of new cases annually is over 500,000, and the yearly incidence comprises between 2.5 and 7% of patients with liver cirrhosis. The incidence varies between different geographic areas, being higher in developing areas; males are predominantly affected, with a 2:3 male/female ratio. The heterogeneous geographic distribution reflects the epidemiologic impact of the main etiologic factors and environmental risk, which are the hepatitis B (HBV) and hepatitis C (HCV) viruses. The percentage of cases of hepatocellular carcinoma attributable to HBV worldwide is 52.3% and is higher in Asia where the seroprevalence of HBsAg in the population is high. However, the vaccination campaign against this virus in some eastern countries has tended to lower the incidence of new cases of hepatocellular carcinoma. The percentage of cases of hepatocellular carcinoma attributable to HCV is 25%, and it is more prevalent in Japan, Spain, and Italy where the association between hepatocellular carcinoma and antibodies to HCV ranges between 50 and 70%. In most cases hepatocellular carcinoma develops in cirrhotic livers, where the persistent proliferation of liver cells represents the key factor of progression to hepatocellular carcinoma independent of the etiology. Another minor risk factor is aflatoxin B1 consumption, which is responsible for most cases of hepatocellular carcinoma in Africa, where the consumption of contaminated foods is common. Other known risk factors are some hereditary diseases, such as hemochromatosis, porphyria cutanea tarda, hereditary tyrosinemia, and alpha1 anti-trypsin deficiency. The natural history of hepatocellular carcinoma is heterogeneous and is influenced by nodule dimension, the mono- or plurifocality of lesions at diagnosis, the growth rate of the tumor, and the stage of the underlying cirrhosis. Available data to date suggest that tumor growth in a cirrhotic liver is variable and that the time in which a lesion in undetectable until it becomes 2 cm is between 4 and 12 months. Therefore, the suggested interval for surveillance screening with ultrasound in patients with liver cirrhosis has been set at 6 months. Patients who should benefit from screening programs are those who would be treated with curative therapy if diagnosed with hepatocellular carcinoma. Thus, the ideal target population should be limited to Child-Pugh's class A cirrhotic patients without significant comorbidity. PMID- 12095925 TI - Genetic alterations and oncogenic pathways in hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is a major type of primary liver cancer and one of the rare human neoplasms etiologically linked to viral factors. Chronic infections with the hepatitis B virus (HBV) and the hepatitis C virus (HCV) have been implicated in about 80% of cases worldwide, and other known environmental risk factors, including alcohol abuse and dietary intake of aflatoxin B1, might synergize with viral infections. Recent insight into the molecular mechanisms leading to HCC development has been provided by the identification of major genetic abnormalities revealed by genomewide allelotype studies and molecular cytogenetic analysis. Moreover, several oncogenic pathways have been implicated in malignant transformation of liver cells. Inactivation of the p53 tumor suppressor gene by mutations and allelic deletions in about 30% of HCC cases has been associated predominantly with exposure to aflatoxin B1 and HBV infection. By contrast, a mutation in the beta-catenin gene in around 22% of HCCs is more rare in HBV-associated tumors. Activation of cyclin D1 and disruption of the Rb pathway are also commonly involved in liver tumorigenesis. New major challenges include the identification of candidate genes located in frequently altered chromosomal regions and that of oncogenic pathways driven by different risk factors. This search might shed some light on the tumorigenic role of HBV and HCV. It might also permit accurate evaluation of major targets for prognostic and therapeutic intervention. PMID- 12095926 TI - Hepatocellular carcinoma: role of estrogen receptors in the liver. AB - Experimental and clinical evidence indicates that estrogens have a relevant role in the pathogenesis of cancer of hormone-sensitive organs. Estrogen receptors (ERs) are present in liver cells. Normal liver expresses almost exclusively wild type ERs derived from the full-length transcript of the gene. During progression of liver disease to hepatocellular carcinoma, variant forms of ERs have been demonstrated that greatly influence the course of the disease and the possibility of palliative treatment. Peritumoral cirrhotic tissue of patients with hepatocellular carcinoma, especially males, expresses a variant form of ER (vER) with an exon 5 deletion. In hepatocellular carcinoma, vER largely predominates and sometimes becomes the only form expressed. That the occurrence of vER alone is limited almost exclusively to males suggests that it could be one of the molecular events that eventually lead to the preferential development of hepatocellular carcinoma in males. In addition, the presence of vER appears most frequently in patients infected with the hepatitis B virus. The growth rate of hepatocellular carcinoma in patients with vER is also significantly higher than that in patients with tumors expressing wtER. PMID- 12095927 TI - Circulating IL-6 and sIL-6R in patients with hepatocellular carcinoma. AB - Interleukin-6 plays a central role in regulating the immune system, hematopoiesis, and acute phase reaction. It interacts with a receptor complex consisting of a specific ligand-binding protein (IL-6R, gp80) and a signal transduction protein (gp130). In this report, serum levels of IL-6 and a soluble form of the interleukin-6 receptor (sIL-6R) were evaluated in patients with hepatocellular carcinoma. The correlation between IL-6 and sIL-6R values, the stage of hepatocellular carcinoma, and main liver function tests was also studied. PMID- 12095928 TI - Expression of HIP/PAP mRNA in human hepatoma cell lines. AB - The present study attempts to shed more light on the role of hepatocarcinoma intestine-pancreas/pancreatic associated protein (HIP/PAP) in hepatoma cells. We initially examined, by reverse transcription-polymerase chain reaction (RT-PCR), the HIP/PAP transcripts present in human hepatoma cell lines of different origins and with different grades of differentiation and genetic profiles. We also used DNA sequencing analysis to investigate the structure of the HIP/PAP gene. Further investigation is necessary to define the role of HIP/PAP during the development of human hepatocellular carcinoma and to ascertain whether the use of different transcripts is helpful in regulating HIP/PAP expression in transformed liver cells. PMID- 12095929 TI - Neuroblastoma: a challenge for pediatric oncology of the third millennium. AB - Neuroblastoma is the only cancer of childhood considered in this conference on hormone-related tumors, because the percentage of deaths in children with this rare type of cancer is still very high. Pediatricians feel the need for help from basic researchers to better understand the biological nature of this disease and to improve protocols and the challenge of cure. PMID- 12095930 TI - Amplified MYCN in human neuroblastoma: paradigm for the translation of molecular genetics to clinical oncology. AB - Increase in the dosage of cellular oncogenes by DNA amplification is a frequent genetic alteration of cancer cells. In neuroblastoma, amplification of the gene MYCN has been associated with aggressively growing cancers and is an indicator for poor prognosis. MYCN amplification is of predictive value in identifying patients with neuroblastoma who require specific therapeutic regimens and those who do not benefit from chemotherapy. PMID- 12095931 TI - Linkage analysis in families with recurrent neuroblastoma. AB - Neuroblastoma is a neural crest-derived tumor of childhood with a serious prognosis; only 20% of patients with stage 4 disease survive 5 years from diagnosis. Mechanisms involved in neuroblastoma development are unclear, but the engagement of many neuroblastoma-related gene(s) is suggested by specific chromosomal alterations. Most prominent among these is the amplification of the MYCN oncogene and the deletion of the 1p36 region. Other genetic aberrations have been discovered over the years such as deletions of 11q and 14q and gain of 17q. Although tumor aggressiveness greatly depends on the most frequent genetic abnormalities, to date no neuroblastoma-related gene has been discovered. Neuroblastoma usually occurs sporadically, but 1.5% of all diagnosed cases show familial recurrence with an autosomal dominant inheritance and incomplete penetrance. A comparison between hereditary and sporadic neuroblastomas led Knudson and Strong to gather that the two-hit hypothesis, proposed for retinoblastoma, could be applied to neuroblastoma. To determine if the 1p36 region harbors a predisposition gene for familial neuroblastoma, we carried out linkage analysis at 1p36 loci in two families with recurrent neuroblastoma. Similarly, we analyzed loci of chromosome 16, where a predisposition locus was recently mapped. We also analyzed markers located close to several candidate genes (RET, NF1, GDNF, GFRA1, EDNRB, and EDN3) involved to a different extent in other neurocristopathies. Our findings indicate that the candidate chromosomal regions and genes analyzed are not in linkage with neuroblastoma. PMID- 12095932 TI - Breast cancer registry in Palermo and its Province: incidence in 1999. AB - The incidence of breast cancer in the city of Palermo and its Province was investigated. The cancer rate was higher in the city of Palermo (100.8/100,000/year), a great southern urban area, than in the 81 municipalities of the Province (79.2/100,000/year). Rates were also compared with those in other geographic areas of Italy, showing a smaller than expected negative north-south gradient in incidence, especially in the young age group, as shown by the cumulative risk observed in the 0-54-year-old group. These findings confirm the role of recent life style changes in the cancer risk distribution. PMID- 12095933 TI - New approaches to breast cancer: oxaliplatin combined with 5-fluorouracil and folinic acid in pretreated advanced breast cancer patients: preliminary reports. AB - Oxaliplatin is a platinum compound that inhibits DNA synthesis. This drug has a broad spectrum of antineoplastic activity, and its results in breast cancer are promising. We began a phase II study in pretreated advanced breast cancer patients using oxaliplatin together with 5-fluorouracil and folinic acid, a combination based on the efficacy of both drugs in breast cancer and their different toxicity profiles. Seventeen patients with advanced breast cancer were treated with oxaliplatin, 5-fluorouracil, and folinic acid, and preliminary data were analyzed. The mean number of courses per patient was 2.82 (range 1-8). The main toxicity was gastrointestinal, with nausea and vomiting G2-3 in 53% of the patients. Hematologic toxicity was moderate with neutropenia G2-3 in 13% of the patients. Among 10 evaluable patients we obtained partial response in one and stabilized the disease in two patients. No data on survival were evaluated. The small number of enrolled and evaluable patients does not permit any conclusions to be drawn. The study is ongoing. PMID- 12095934 TI - Ligand binding and cytochemical analysis of estrogen and progesterone receptors in relation to follow-up in patients with breast cancer. AB - Soluble and nuclear estrogen receptor (ER) content was measured by ligand binding assay, and estrogen and progesterone receptors by immunohistochemical assays (ER ICA and PR-ICA) in 214 patients with breast cancer recruited at the "M. Ascoli" Cancer Hospital Centre in Palermo, Sicily, to assess the discriminant and predictive value of these parameters. On follow-up, data from both ER-ICA and PR ICA showed a statistically significant difference, PR-positive patients having longer disease-free (DSF) and overall (OS) survival than PR-negative ones. Conversely, ER status did not correlate significantly with both DFS (P = 0.6) and OS (P = 0.2). In particular, PR-positive patients had 59 +/- 18 months DFS and 67 +/- 12 months OS, compared to 51 +/- 22 months DFS and 57 +/- 17 months OS of PR negative cases. The present evidence implies that a PR-negative status identifies breast cancer patients with early relapse, as also suggested by previous studies. It also agrees with the results of ligand binding assay of ER, where ER status is a good discriminant and predictor of response to endocrine treatment, but is unable to anticipate early relapse in breast cancer patients. Evidence that PR status is a statistically significant prognostic indicator deserves further study to ascertain whether or not PR should be regarded as an ER-dependent parameter or be related to other biological variables such as growth factor (e.g., EGF), oncogene (e.g., Her2/Neu), or tumor suppressor gene (e.g., p53) products. PMID- 12095935 TI - Modulation of epidermal growth factor receptor in endocrine-resistant, estrogen receptor-positive breast cancer. AB - An increasing body of evidence demonstrates that growth factor networks are highly interactive with estrogen receptor signaling in the control of breast cancer growth. As such, tumor responses to antihormones are likely to be a composite of the estrogen receptor and growth factor inhibitory activity of these agents. The modulation of growth factor networks during endocrine response is examined, and in vitro and clinical evidence is presented that epidermal growth factor receptor signaling, maintained in either an estrogen receptor-dependent or a receptor-independent manner, is critical to antihormone-resistant breast cancer cell growth. The considerable potential of the epidermal growth factor receptor selective tyrosine kinase inhibitor Iressa (ZD 1839) to efficiently treat, and perhaps even prevent, endocrine-resistant breast cancer is highlighted. PMID- 12095936 TI - Molecular mechanisms of RET activation in human cancer. AB - Mutations that produce oncogenes with dominant gain of function target receptor protein tyrosine kinases (PTKs) in cancer and confer uncontrolled proliferation, impaired differentiation, or unrestrained survival to the cancer cell. However, insufficient PTK signaling may be responsible for developmental diseases. Gain of function of the RET receptor PTK is associated with human cancer. At the germline level, point mutations of RET are responsible for multiple endocrine neoplasia type 2 (MEN2A, MEN2B, and FMTC). Mutations of extracellular cysteines are found in MEN2A patients, and a Met918Thr mutation is responsible for most MEN2B cases. At the somatic level, gene rearrangements juxtaposing the tyrosine kinase domain of RET to heterologous gene partners are found in papillary carcinomas of the thyroid. These rearrangements generate the chimeric RET/PTC oncogenes. Both MEN2 mutations and PTC gene rearrangements potentiate the intrinsic tyrosine kinase activity of RET and, ultimately, the RET downstream signaling events. A multidocking site of the C-tail of RET is essential for both mitogenic and survival RET signaling. Such a site is involved in the recruitment of several intracellular molecules, such as the Shc, FRS2, IRS1, Gab1/2, and Enigma. The different activating mutations not only potentiate the enzymatic activity of the RET kinase but also may alter qualitatively RET signaling properties by: (1) altering RET autophosphorylation (in the case of the MEN2B mutation), (2) modifying the subcellular distribution of the active kinase, and (3) providing the active kinase with a scaffold for novel protein-protein interactions (as in the case of RET/PTC oncoproteins). This review describes the molecular mechanisms by which the different genetic alterations cause the conversion of RET into a dominant transforming oncogene. PMID- 12095938 TI - Antiestrogenic regulation of transforming growth factor beta receptors I and II in human breast cancer cells. AB - To obtain more information about the interactions between antiestrogens and transforming growth factor beta (TGF-beta) signaling, we analyzed the influence of different types of antiestrogens on the expression of TGF-beta receptors I (TbetaRI) and II (TbetaRII). Our results show that antiestrogens selectively induce TbetaRII but not TbetaRI mRNA. PMID- 12095937 TI - Proteomic patterns of cultured breast cancer cells and epithelial mammary cells. AB - Breast cancer is one of the leading causes of death from cancer among women in western countries. The different types of breast cancer are grouped into invasive and noninvasive forms. Among the invasive types, ductal infiltrating carcinoma (DIC) is the most common and aggressive form. Using an in vitro model consisting of a DIC-derived cell line (8701-BC) and a nontumoral mammary epithelial cell line (HB2), we used the proteomics approach to search for homology and differences in protein expression patterns between tumoral and nontumoral phenotypes. Within an analysis window comprising 1,750 discernible spots we have currently catalogued 140 protein spots of potential interest. Fifty-eight of them were identified by gel matching with reference maps, immunodetection, or N terminal microsequencing and classified into four functional groups. Twelve proteins were found differentially expressed in two cell lines: four were uniquely present in the neoplastic cell proteome and eight in epithelial cells. In addition, 53 proteins displayed different relative expression levels between the two cell lines, that is, 44 were more elevated in cancer cells and 9 in HB2 cells. Among proteins with greater relative abundance in cancer cells we identified glycolytic enzymes (or their isoforms), which may indicate that the known metabolic dysregulation in cancer can reflect oncogenic-related defects of glycolytic gene expression. PMID- 12095939 TI - Objective response to treatment as a potential surrogate marker of survival in breast cancer. AB - Until recently, objective tumor response to chemotherapy was used as the primary endpoint in phase II trials aimed at assessing the anti-tumor activity of new drugs. However, it was not accepted as a surrogate endpoint of survival for efficacy trials in solid tumors, and it was not believed to be associated with survival benefit. The recent demonstration that objective response is indeed a valid surrogate endpoint of survival in colorectal cancer, together with strong indirect evidence supporting a similar role of response in breast cancer, opens new possibilities for both the design of trials in metastatic breast cancer and the clinical decision in individual patients with the disease. PMID- 12095940 TI - Nutrition and prostate cancer: a review. AB - Despite intense efforts, little is known about the etiology of prostate cancer, and treatment of advanced forms of the disease has had limited success. Nonetheless, epidemiologic studies combined with animal model and in vitro experiments indicate that natural components of the diet, including n-3 PUFA, the carotenoid lycopene, and the trace element selenium, may serve as chemopreventive agents that suppress the growth and dissemination of neoplastic prostate cells. Until further study, however, soy isoflavones should be viewed with some caution, especially as adjuvant's to chemotherapy, in patients with hormone-refractory prostate cancer. Future studies, using different forms and doses of selenium and tomato carotenoids, may shed new light on the etiology and prevention of prostate cancer. PMID- 12095941 TI - Intercellular communication and human prostate carcinogenesis. AB - Gap-junction-mediated intercellular communication (GJIC) is required for completion of embryonic development, tissue homeostasis, and regulation of cell proliferation and death. Although, as emphasized in several reports, defects or disruption of GJIC may be important in carcinogenesis, the potential role of GJIC in the onset and progression of human prostate cancer remains ill-defined. The gap junction channel-forming connexins (Cx) comprise a multigene family of highly conserved proteins that are differentially expressed in a tissue- and development specific manner; changes in connexin expression are also commonly seen during cellular differentiation. However, when multiple connexins are concurrently expressed, gap junction channels may consist of more than one connexin species. This is important, because only certain pairings give rise to functional channels. In our studies, we investigated GJIC in a panel of both nontumorigenic (RWPE-1) and malignant (RWPE-2, LNCaP, DU-145) human prostate epithelial cells, compared to a normal rat liver epithelial F344 (WB-1) cell line, as it was found to be junctionally proficient. In addition, expression and regulation of Cx43 and Cx32 were also inspected using western blot analysis. The ability of hormones, antihormones, and the antihypertensive drug forskolin to restore GJIC in nontumorigenic and malignant human prostate epithelial cells was examined by the scrape-loading/dye transfer (SL/DT) or fluorescence recovery after photobleaching (FRAP) methods using an Ultima laser cytometer. Results from both assays showed that neither nontumorigenic nor malignant prostate cells have functional GJIC. However, both estrone (E1) and forskolin (FK) induced a significant increase (4.4 and 2.8-fold, respectively) in cell-cell communication only in the RWPE-1 cells. Interestingly, the use of Matrigel, a solubilized basement membrane, as substrate for cell attachment and growth resulted in the rescue of GJIC activity in RWPE-1 cells, as revealed by the SL/DT method. Furthermore, E1 induced a twofold increase in connexin 43 (Cx43), whereas forskolin caused a 50% reduction in Cx32 expression in RWPE-1 cells. These data suggest that agents that increase Cx43:Cx32 ratio may restore GJIC in junctionally deficient cells, providing a basis for the development of new strategies for the prevention and treatment of human prostate cancer. PMID- 12095942 TI - Precancerous lesions and conditions of the prostate: from morphological and biological characterization to chemoprevention. AB - Prostatic intraepithelial neoplasia (PIN) is composed of dysplastic cells with a luminal cell phenotype, expressing the androgen receptor as well as prostate specific antigen. PIN is characterized by progressive abnormalities of phenotype that are intermediate between normal prostatic epithelium and cancer, indicating impairment of cell differentiation and regulatory control with advancing stages of carcinogenesis. High-grade PIN is considered the most likely precursor of prostatic carcinoma, according to virtually all available evidence. Androgen deprivation decreases the prevalence and extent of PIN and the degree of capillary vascularization (e.g., angiogenesis) in the surrounding stroma via suppression of vascular endothelial growth factor production. Prostatic carcinoma is also likely to arise from precursor lesions other than high-grade PIN such as low-grade PIN, atypical adenomatous hyperplasia, malignancy-associated foci, and atrophy. PMID- 12095943 TI - Src is an initial target of sex steroid hormone action. AB - Recent observations that steroids use pathways universally known to be regulated by growth factors and interleukins highlight the following points: (1) Steroid stimulation of the canonical pathway Src/Ras/Erk signaling from membrane to nuclei or its single members has been observed in different cell types including human cancer-derived cells, neurons, osteoblasts, osteocytes, and endothelial cells. This stimulation has been reconstituted and analyzed in transiently transfected cells. (2) Cellular context and intracellular localization of receptors are crucial in determining the biological effects evoked by this hormonal stimulation: proliferation, protection from apoptosis, and vasorelaxation. (3) Classical steroid receptors localized in the extranuclear compartment directly and, in some cases, simultaneously interact with Src. They are capable of unexpected cross talks responsible for the observed effects. (4) Other signaling pathways including P13K/AKT are also stimulated by steroids. The aim of future work will be to arrive at an integrated general view of the different signaling pathways activated by steroids and to analyze the concert between these pathways and the hormonal transcriptional action. This general view should be simultaneously verified in different cell contexts, under different physiologic and pathologic conditions. We expect that the new technologies, above all gene and protein microarray, will make this goal feasible. PMID- 12095944 TI - From castration-induced apoptosis of prostatic epithelium to the use of apoptotic genes in the treatment of prostate cancer. AB - Current knowledge of the mechanisms regulating androgen-ablation-induced apoptosis is reviewed, and our efforts to develop a system in which genes of the apoptotic pathway are used to induce therapeutic apoptosis in experimental models of prostate cancer are described. PMID- 12095945 TI - Biological selection criteria for radical prostatectomy. AB - Tumors clinically confined to the prostate gland (T1-2) are heterogeneous with respect to pathological staging and outcome after definitive radical surgery (radical prostatectomy). The preoperative prognostic factors that could predict pathological stage and outcome of individual patients with clinically localized prostate cancer are reviewed. New preoperative factors have been identified by histological analysis of needle biopsy prostate specimens in addition to Gleason grading score, serum markers (PSA), and clinical staging. These factors are related to tumor volume, zonal origin of the tumor, and spread into the gland and surrounding tissues. Other biological factors are identified by molecular and immunohistochemical analysis (neuroendocrine differentiation, DNA content, microvessel density, and perineural invasion). Biomolecular factors can also be assessed preoperatively on serum samples (free/total PSA ratio, PSA RT-PCR). Although only a few of these factors have a role in predicting treatment failure and/or disease recurrence, the neural network analysis seems to be the most important tool for identifying patients with more aggressive disease. A combination of these new factors, also using neural networks, could be relevant in the preoperative management of patients with prostate cancer to identify those with confined disease and to select those suitable for a "nerve sparing radical prostatectomy" to preserve sexual function and to achieve definitive cancer control. PMID- 12095946 TI - Connexin expression in nonneoplastic human prostate epithelial cells. AB - Expression of gap-junction proteins connexins (Cx), specifically Cx43, Cx32, and Cx26, in both nontumorigenic (RWPE-1) and tumorigenic (RWPE-2) human prostate epithelial cells as well as in two cell clones (WPEI-7 and WPEI-10) originating from the RWPE-1 cell line was investigated. The aim was to determine whether individual connexins are differentially expressed in cultured cells. Western blot analysis revealed striking differences in the expression of individual connexins in the cell lines studied. In particular, Cx43 is largely expressed in RWPE-1 and WPEI-10 cells, whereas Cx32 is expressed predominantly in RWPE-2 and WPEI-7 cells. In addition, both forskolin and estrone increase Cx43 expression levels in WPEI-10 cells, with no apparent effect on WPEI-7 cells. Conversely, forskolin and especially estrone induce a marked increase of Cx32 in WPEI-7 cells, whereas Cx32 expression is limitedly affected by both agents in WPEI-10 cells. Overall, expression levels of Cx43 and Cx32 appear to be inversely related, with RWPE-1 and WPEI-10 cells having a significantly higher Cx43 to Cx32 ratio than that observed in RWPE-2 and WPEI-7 cells. We recently reported that junctional communication could be rescued in RWPE-1 cells by either forskolin or estrone and that restoration of GJIC is associated with an increase of Cx43 or a decrease of Cx32, or both, eventually leading to a marked rise of the Cx43 to Cx32 ratio. Studies are currently ongoing in our laboratories to assess the potential effect of agents increasing the Cx43 to Cx32 ratio on GJIC activity in these systems. PMID- 12095947 TI - Methods to obtain more clinical and pathologic information from needle core biopsy of the prostate gland. AB - Prostate needle biopsy can disclose important clinical information on tumor extension and grading, useful prognostic parameters for therapeutic choices and prognostic definition. To obtain more histopathologic information on specimens and clinical prognostic parameters, we used a new method to handle and embed tissue fragments of prostate biopsy as well as a new trans-rectal probe (TRUS) with an end-fire convex ultrasound transducer to guide ultrasonically precise needle placement in the prostate gland. In our experience these methods combined can be useful in obtaining reliable clinical and prognostic information in the management of patients with prostate cancer. PMID- 12095948 TI - Human type 1 estrogen sulfotransferase: catecholestrogen metabolism and potential involvement in cancer promotion. AB - Using purified human type 1 estrogen sulfotransferase (hEST1), we show that the best substrate for this enzyme is 2-hydroxy-catecholestrogen. The enzyme also catalyzes the transformation of 4-hydroxy-estrogens and 16-hydroxy-estrogens, but with a lower affinity. We also present evidence to indicate that estrogen sulfotransferase may play a role in processes other than the detoxification and elimination of steroids. Indeed, hEST1 may also be involved in the production of stable precursors for local steroid biosynthesis or in the activation of promutagenic estrogen metabolites into carcinogens. PMID- 12095949 TI - Prevention and treatment of breast cancer by suppressing aromatase activity and expression. AB - Estrogen promotes the proliferation of breast cancer cells. Aromatase is the enzyme that converts androgen to estrogen. In tumors, expression of aromatase is upregulated compared to that of surrounding noncancerous tissue. Tumor aromatase is thought to stimulate breast cancer growth in both an autocrine and a paracrine manner. A treatment strategy for breast cancer is to abolish in situ estrogen formation with aromatase inhibitors. In addition, aromatase suppression in postmenapausal women is being evaluated as a potential chemopreventive modality against breast cancer. One area of aromatase research in this laboratory is the identification of foods and dietary compounds that can suppress aromatase activity. In vitro and in vivo studies have found that grapes and mushrooms contain chemicals that can inhibit aromatase. Therefore, a diet that includes grapes and mushrooms would be considered preventative against breast cancer. Another area of our aromatase research is the elucidation of the regulatory mechanism of aromatase expression in breast cancer tissue. Increased aromatase expression in breast tumors is attributed to changes in the transcriptional control of aromatase expression. Whereas promoter I.4 is the main promoter that controls aromatase expression in noncancerous breast tissue, promoters II and I.3 are the dominant promoters that drive aromatase expression in breast cancer tissue. Our recent gene regulation studies revealed that in cancerous versus normal tissue, several positive regulatory proteins (e.g., nuclear receptors and CREB1) are present at higher levels and several negative regulatory proteins (e.g., snail and slug proteins) are present at lower levels. This may explain why the activity of promoters II and I.3 is upregulated in cancerous tissue. In addition, our in vitro transcription/translation analysis using plasmids containing T7 promoter and the human snail gene as a reporter capped with different untranslated exon Is revealed that exon PII-containing transcripts were translated more effectively than were exon I.3-containing transcripts. An understanding of the molecular mechanisms of aromatase expression between noncancerous and cancerous breast tissue, at both transcriptional and translational levels, may help in the design of a therapy based on suppressing aromatase expression in breast cancer tissue. PMID- 12095951 TI - Diet and breast cancer. AB - The preponderance of evidence suggests a role for fat and alcohol as risk factors for breast cancer. The role of milk is more controversial with some studies suggesting that milk is a risk factor and others that consumption of milk is protective against breast cancer. No other major nutrient appears to play a significant role in increasing breast cancer risk. On the other hand, there is increasing evidence that a variety of micronutrients and hormones appear to have significant anticancer activity. These range from steroids such as dehydroepiandrosterone (DHEA) and its analysis to indoles, isothiocyanates, and isoflavone derivatives. These compounds act directly by interfering with cyclins and promoting apoptosis as well as indirectly by altering estrogen metabolism in a favorable direction. These effects are not merely theoretical actions in cell culture and tissue explants; they have been demonstrated in human patients as a range of studies have demonstrated. PMID- 12095950 TI - Anti-aromatase chemicals in red wine. AB - Estrogen synthesized in situ plays a more important role in breast cancer cell proliferation than does circulating estrogen. Aromatase is the enzyme that converts androgen to estrogen and is expressed at a higher level in breast cancer tissue than in surrounding noncancer tissue. A promising route of chemoprevention against breast cancer may be through the suppression of in situ estrogen formation using aromatase inhibitors. A diet high in fruits and vegetables may reduce the incidence of breast cancer, because they contain phytochemicals that can act as aromatase inhibitors. In our previous studies, we found that grapes and wine contain potent phytochemicals that can inhibit aromatase. We show that red wine was more effective than white wine in suppressing aromatase activity. Interestingly, our results from white wine studies suggest a weak inductive effect of alcohol on aromatase activity. On the other hand, the potent effect of anti-aromatase chemicals in red wine overcomes the weak inductive effect of alcohol in wine. Several purification procedures were performed on whole red wine to separate active aromatase inhibitors from non-active compounds. These techniques included liquid-liquid extraction, silica gel chromatography, various solid phase extraction (SPE) columns, and high performance liquid chromatography. An active Pinot Noir red wine SPE C18 column fraction (20% acetonitrile:water) was more effective than complete Pinot Noir wine in suppressing aromatase assay. This red wine extract was further analyzed in a transgenic mouse model in which aromatase was over-expressed in mammary tissue. Our gavaged red wine extract completely abrogated aromatase-induced hyperplasia and other neoplastic changes in mammary tissue. These results suggest that red wine or red wine extract may be a chemopreventive diet supplement for postmenopausal women who have a high risk of breast cancer. Further research is underway to purify and characterize the active compounds in red wine that are responsible for the inhibition of aromatase. PMID- 12095952 TI - Effects of weight control and physical activity in cancer prevention: role of endogenous hormone metabolism. AB - Excess body weight and/or lack of physical activity are increasingly recognized as major risk factors for cancer of the colon, breast, endometrium, and prostate. This paper reviews the effects of excess body weight and physical inactivity on endogenous hormone metabolism (insulin, the IGF-I/IGFBP system, and sex steroids) and of endocrine alterations with risk of cancer of the endometrium, breast, prostate, and colon. PMID- 12095953 TI - The Mediet Project. AB - Preliminary evidence from a case control study of healthy postmenopausal women living in Palermo, Sicily, is presented to investigate the potential impact of a traditional Mediterranean diet on the risk of developing breast cancer. Of the 230 women who fulfilled specific eligibility criteria, 115 were enrolled in the study based on serum testosterone values equal to or greater than the median population value (0.14 microg/ml). Women were then individually randomized into a diet intervention (n = 58) and a control (n = 55) group. Women in the intervention group attended a weekly "cooking course" for 1 year, being trained by professional chefs in the correct use of the natural ingredients of the traditional Mediterranean diet, including whole cereals, legumes, seeds, fish, cruciferous vegetables, and many others. The intervention group was subsequently instructed to follow the learned diet at home, while the control group was only advised to increase the consumption of fruits and vegetables, as recommended by WHO. The following measures were taken at the beginning, middle, and end of the study: (a) fasting blood and 12-hour urine samples to assay defined hormonal endpoints; (b) height, weight, and circumference of the waist and hip; and (c) a food frequency and computerized 24-hour dietary recall questionnaire. After 1 year, both the control and the intervention groups showed satisfactory compliance rates (81 and 85%, respectively). In addition, preliminary results so far obtained reveal an unequivocal trend towards weight loss, a strong reduction in cholesterol levels, and a psychophysical feeling of well-being by women adopting the Mediterranean diet. The study is currently ongoing to verify the association of changes in serum and urine hormone levels and breast cancer risk in the intervention group. PMID- 12095954 TI - Murine models of paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal disorder characterized by chronic intravascular hemolysis, cytopenia, and an increased tendency to thrombosis. All patients with PNH studied so far have a somatic mutation of phosphatidyl inositol glycan complementation group A (PIG-A), an X linked gene involved initially in the biosynthesis of the glycosyl phosphatidylinositol (GPI) molecule, which serves as an anchor for many cell surface proteins. The mutation occurs in a hematopoietic stem cell, and consequently, all cells derived from the mutated stem cell are devoid of GPI linked proteins. The absence of GPI-linked proteins explains some clinical symptoms of the disease but not the mechanism that allows the expansion of the mutated clone. By using targeted disruption of the PIG-A gene in mouse embryonic stem cells, some mice models of PNH have been generated. These animals have a discrete proportion of blood cells devoid of GPI-linked proteins, and although not anemic, they have evidence of hemolysis. The clinical course of these animals is benign, and there are no signs of a substantial expansion of the PNH clone, as observed in human patients. The fact that these animals do not develop the disease strongly supports the notion that a mutation of PIG-A is not sufficient per se to cause PNH and that another factor, namely, bone marrow failure, is necessary to allow proliferation and expansion of the PNH clone. PMID- 12095955 TI - Molecular genetics of acute myeloid leukemia. AB - Recurring chromosomal abnormalities are detected in most patients with acute myeloid leukemia (AML). They may be associated with a distinct AML FAB subtype or may identify distinct clinicobiological entities within the same FAB subtype. Therefore, cytogenetic investigation has a pivotal role in AML diagnosis. In addition, it is one of the most valuable prognostic determinants of the disease, as recently demonstrated. The development of new molecular techniques, such as reverse transcriptase polymerase chain reaction and fluorescence in situ hybridization, has allowed perfect definition of the chromosome regions containing genes with a crucial role in normal hemopoiesis and leukemia. Understanding the action of such genes provides new insights into AML pathogenesis and has led us to envisage new therapeutic options. PMID- 12095956 TI - From genes to therapy: the case of Philadelphia chromosome-positive leukemias. AB - The Philadelphia chromosome (Ph-chromosome) has long represented the only cytogenetic abnormality known to be associated with a specific malignant disease in humans, being present in more than 95% of patients with chronic myelogenous leukemia. This abnormality is the result of a reciprocal translocation between the long arms of chromosome 9 and 22, t(9;22)(q34;q11), and its presence is not restricted to chronic myelogenous leukemia, but can also be found in 30% of cases of acute lymphoblastic leukemia in adults. In the 1980s, the molecular counterpart of the chromosomal rearrangement was identified to consist of the juxtaposition of parts of the BCR and ABL genes to form a BCR-ABL hybrid gene. The resulting chimeric proteins (P210 and P190), which retain constitutively activated tyrosine kinase activity, have demonstrated a causative role in the genesis of the leukemic process. Although many aspects of the BCR-ABL driven transformation remain unsolved, great advances in understanding the molecular pathology of Ph-positive leukemias resulted in meaningful improvement in the clinical setting. Molecular tools to diagnose disease (PCR, FISH, and southern blot) and to monitor minimal residual disease after potential curative treatment are now in current practice, and new powerful therapeutic tools have emerged that target the molecular oncogenic pathways activated in Ph-positive cells. Among them, specific ABL tyrosine kinase inhibitors recently obtained extraordinary results in many clinical protocols. This review summarizes the most recent advances in this field with special focus on the putative mechanisms of the transformation and progression of chronic myelogenous leukemia and on the major impact that understanding the molecular biology of these diseases is having in clinical practice. PMID- 12095957 TI - Multidimensional flow cytometry immunophenotyping of hematologic malignancy. AB - Immunophenotyping of hematologic malignancies represents one of the most relevant clinical applications of flow cytometry. Classically, leukemic/lymphomatous cells have been considered to reflect the immunophenotypic characteristics of different precursors and mature healthy cells blocked at certain differentiation stages. Recently, accumulating evidence has shown that neoplastic cells display several aberrant phenotypic patterns. These aberrant phenotypes are believed to reflect genetic abnormalities present in pathologic cells, and recent data have shown that at least in acute leukemias, myelodysplastic syndromes, chronic lymphoproliferative disorders, and plasma cell dyscrasias, they may be present in almost all patients. The aim of this work is to review recent advances in flow cytometry and the role of gating strategies more useful in the identification and characterization of neoplastic cells of different hematologic malignancies. PMID- 12095958 TI - Androgen receptor status in nontumoral and malignant human colorectal tissues. AB - Data on androgen receptor (AR) status of nontumoral and malignant human colorectal tissues are compared using ligand binding assay in 22 patients who underwent surgery for colorectal cancer at the "M. Ascoli" Cancer Hospital Centre in Palermo, Sicily. In nontumoral tissues, ARs were predominantly (67%) positive, with 25% of cases having a 0/+ status. Conversely, malignant tissues showed only 32% of cases with a positive (+/+) AR status, with a proportional increase of 0/+ cases (from 25% to 55%); the extent of AR-negative (0/0) cases remained fairly constant (8-9%). Overall, our evidence indicates that nontumoral colorectal tissues have a predominantly positive (+/+) AR status and that this condition shifts towards a significant decrease of AR-positive cases in cancer tissues. Studies on the relation between status of sex steroid receptors and specific biomolecular markers in human colorectal tumors are currently being carried out in our laboratories. PMID- 12095959 TI - Correlation between Helicobacter pylori infection and IL-18 mRNA expression in human gastric biopsy specimens. AB - Our data indicate that H. pylori infection is associated with active interleukin 18 production in patients with chronic gastritis. Different cell types appear to be involved in this activity and may play a role in the development of immunopathologic damage. PMID- 12095960 TI - p53 and anti-p53 antibodies as possible markers of a switch towards a neoplastic phenotype in patients infected by Helicobacter pylori. AB - Helicobacter pylori is a definite carcinogen whose mechanism of action is still unknown. The aim of this work was (1) to determine the presence of p53 protein and related antibodies in patients affected by various gastric pathologies and chronically infected with H. pylori, and (2) to try to discover a test to be used as a marker of a possible switch towards a neoplastic phenotype. PMID- 12095961 TI - Cytokine induction in murine bladder tissue by type 1 fimbriated Escherichia coli. AB - The local cytokine response to uropathogenic phenotype Escherichia coli KBC211 infection exhibits characteristics of both TH1 and TH2 profiles. Interleukin (IL) 6, MIP-2, IL-12, IL-18, and tumor necrosis factor-alpha (TNF-alpha) are expressed, but IL-4, IL-5, and IL-10 are also present at low levels. This is clearly a complex response that should be explored more fully. The relative contributions of the bladder epithelium and other cells of the bladder wall should also be determined. Epithelial cytokine responses may be considerable, and because these cells are the first to encounter the pathogen, they will be of great importance in the immune response to pathogenic E. coli. PMID- 12095963 TI - Honoring Walter Lawrence, Jr., MD: Society of Surgical Oncology President 1979 80. PMID- 12095964 TI - Long-term complications of breast-conservation therapy: can the incidence be reduced? PMID- 12095965 TI - Integrating surgery and radiotherapy to reduce toxicity while maintaining local control for breast cancer: a fine balance. PMID- 12095966 TI - Lymphatic mapping of nodal micrometastasis in colon cancer: putting the cart before the horse? PMID- 12095967 TI - Preoperative chemoradiation for rectal cancer: "Apples & Pears, Tables and Chairs. . .". PMID- 12095968 TI - Preoperative chemoradiation treatment strategies for localized sarcoma. AB - BACKGROUND: Over the past 2 decades, there has been increasing interest in chemoradiation treatment strategies for patients with soft tissue sarcomas. Investigators have evaluated: (1) the optimal route for chemotherapy administration (intra-arterial vs. intravenous); (2) the possible advantages of protracted infusion of the radiosensitizer versus brief infusion; (3) the efficacy and toxicity of various intravenous and oral radiation sensitizers; and (4) the efficacy of sequential versus concurrent combined modality treatment. METHODS: The English-language literature addressing chemoradiation for localized and locally advanced extremity and retroperitoneal sarcomas was reviewed. RESULTS: All studies have been pilot, phase I, or phase II designs. The most commonly used radiosensitizer for concurrent chemoradiation has been doxorubicin, administered intravenously in most recent reports. In the studies that have included assessment of recurrence-free survival, preoperative chemoradiation combined with surgery has resulted in favorable local control rates, often in excess of 90% for patients with localized and locally advanced extremity sarcomas. CONCLUSIONS: The toxicities and recurrence-free outcome with chemoradiation plus surgery for soft tissue sarcoma still need to be compared to these with surgery and pre- or postoperative radiation. However, the generally favorable local control rates reported for chemoradiation justify continued investigation of preoperative chemoradiation strategies for localized sarcoma. PMID- 12095969 TI - Long-term complications associated with breast-conservation surgery and radiotherapy. AB - BACKGROUND: Breast-conservation surgery plus radiotherapy has become the standard of care for early-stage breast cancer; we evaluated its long-term complications. METHODS: We selected patients treated with surgery and radiotherapy between January 1990 and December 1992 (an era in which standard radiation dosages were used) with follow-up for at least 1 year. Patients were prospectively monitored for treatment-related complications. Median follow-up time was 89 months. RESULTS: A total of 294 patients met the selection criteria. Grade 2 or higher late complications were identified in 29 patients and included arm edema in 13 patients, breast skin fibrosis in 12, decreased range of motion in 4, pneumonitis in 2, neuropathy in 2, fat necrosis in 1, and rib fracture in 1. Arm edema was more common after lumpectomy plus axillary node dissection than after lumpectomy alone. Arm edema occurred in 18% of patients who underwent surgery plus irradiation of the lymph nodes and 10% who underwent surgery without nodal irradiation. CONCLUSIONS: Breast-conservation surgery plus radiotherapy was associated with grade 2 or higher complications in only 9.9% of patients. Half of these complications were attributable to axillary dissection, it is hoped that lower complication rates can be achieved with sentinel lymph node biopsy. Breast conservation surgery and radiotherapy is associated with grade 2 or greater complications in only 9.9% of patients. Nearly half of these complications are attributable to axillary dissection. PMID- 12095970 TI - Validation of sentinel node mapping in patients with colon cancer. AB - BACKGROUND: Sentinel lymph node (SLN) mapping techniques have been validated in breast cancer and melanoma. This study summarizes our experience with SLN mapping for colon cancer. METHODS: Fifty-five patients with colon cancer underwent intraoperative SLN mapping. One mL of 1% isosulfan blue was injected subserosally around the tumor. The first nodes highlighted with blue were identified as the SLNs. SLNs underwent multiple sectioning and immunohistochemical staining for cytokeratin. The overall learning curve was calculated. RESULTS: Lymphatic mapping adequately identified at least 1 SLN in 45 patients (82%). SLNs adequately predicted regional status in 44 of 45 (98%) cases. In 9 of 45 cases (20%), the SLNs were the only sites of metastases. Among the 14 cases that were SLN positive, 6 of 55 patients (11%) were positive only by immunohistochemistry. Of the 31 cases with negative SLNs, 1 case had a 3.5-mm pericolonic tumor replaced non-SLN (3% false-negative rate). The overall learning curve stabilized after five cases. CONCLUSIONS: Intraoperative SLN mapping is a feasible technique, with a quick learning curve, and had a reasonable SLN identification rate. Negative SLNs accurately predict the status of non-SLNs 97% of the time. Eleven percent of patients were upstaged by demonstration of micrometastases and may benefit from adjuvant chemotherapy. PMID- 12095971 TI - hMLH1 and hMSH2 gene mutation in Brazilian families with suspected hereditary nonpolyposis colorectal cancer. AB - BACKGROUND: The aim of this study was to search for mutations in the human mutS homolog 2 (hMSH2) and human mutL homolog 1 (hMLH1) genes in 25 unrelated Brazilian kindreds with suspected hereditary nonpolyposis colorectal cancer (HNPCC). METHODS: The families were grouped according to the following clinical criteria: Amsterdam I or II; familial colorectal cancer (CRC); an early age of onset of CRC in the proband only; or with at least one or two relatives who had HNPCC-related cancers; CRC in the proband only. All patients were studied with direct sequencing. RESULTS: Ten mutations were detected (10 of 25 [40%]); of nine different mutations, seven were novel. The hMLH1 gene had a higher mutation detection rate than hMSH2 (8 of 25 [32%] vs. 2 of 25 [8%]). Only 3 of these 10 families fulfilled the Amsterdam criteria. Two different polymorphisms were detected in the hMLH1 gene and four in the hMSH2 gene. CONCLUSIONS: The hMLH1 gene had a higher mutation detection rate than hMSH2. The physician who deals with CRC must take into consideration the heredity issue with patients who present with an early age of onset or a familial history of CRC- or HNPCC-related cancers, including gastric cancer, even if they do not fulfill the former Amsterdam criteria. PMID- 12095972 TI - Prognostic impact of lymphatic and/or blood vessel invasion in patients with node negative advanced gastric cancer. AB - BACKGROUND: Heterogeneous survival rates in patients with similar clinicopathologic characteristics or molecular prognostic markers have been noted. This study was conducted to evaluate the prognostic effect of lymphatic and/or blood vessel invasion (LBVI), identified by routine hematoxylin and eosin staining, on the outcome of patients with node-negative advanced gastric cancer. METHODS: A total of 280 patients who underwent curative gastrectomy for advanced gastric cancer without lymph node metastasis were retrospectively reviewed. Univariate and multivariate analyses of the clinicopathological features, recurrences, and prognoses of patients with and without LBVI were performed. RESULTS: Lymphatic vessel invasion (LVI) was noted in 20.0%, blood vessel invasion (BVI) in 5.4%, and either LVI or BVI in 22.5%. None of the clinicopathologic features was related to LBVI. Patients with LBVI had a recurrence rate of 26.8%, whereas patients without LBVI had a recurrence rate of 13.5% (P =.018). The 5-year survival rates were 82.4% for patients without LBVI and 67.1% for patients with LBVI (P =.0222). LBVI was shown to be an independent risk factor for recurrence (relative risk, 2.30; 95% confidence interval, 1.06 4.99) and poor prognosis (relative risk, 1.88; 95% confidence interval, 1.07 3.29). CONCLUSIONS: LBVI is an adverse prognostic indicator and the presence of LBVI seems to provide useful information for the prognosis and clinical management of patients with node-negative advanced gastric carcinoma. PMID- 12095973 TI - Chemoradiation instead of surgery to treat mid and low rectal tumors: is it safe? AB - BACKGROUND: The main treatment for rectal carcinoma is surgery. Preoperative chemoradiation (CRT) is advocated to reduce local recurrence and improve resection of mid and low tethered rectal tumors. METHODS: Fifty-two patients with mid or low rectal tumors underwent CRT (external beam radiation plus 5 fluorouracil plus folinic acid). Patients who had low rectal tumors with complete response (CR) were not submitted to surgical treatment. All other patients were submitted to surgery, independently of the response. Mean follow-up was 32.1 months. RESULTS: Five-year overall survival was 60.5%. Clinical evaluation after CRT showed CR in 10 cases (19.2%), all low tumors; incomplete response (>50%) in 21 (40.4%); and no response (<50%) in 19 (36.6%). Among the 10 cases with CR, 8 presented with local recurrence within 3.7 to 8.8 months. Two patients were not submitted to surgery and are still alive without cancer after 37 and 58 months. Thirty-nine patients had radical surgery. Seven had local recurrences after CRT plus surgery (17.9%). Overall survival was negatively affected by lymph node metastases (P =.017) and perineural invasion (P =.026). CONCLUSIONS: Exclusive CRT approach is not safe to treat patients with low infiltrative rectal carcinoma. PMID- 12095974 TI - Placement of self-expanding metal stents for acute malignant large-bowel obstruction: a collective review. AB - BACKGROUND: The purpose of this study was to review our experience with self expanding metal stents as the initial interventional approach in the management of acute malignant large-bowel obstruction. METHODS: Twenty-six patients who underwent placement of colonic stents at our institution between June 1994 and June 2000 were identified and reviewed. RESULTS: In 14 patients, the stents were placed for palliation, whereas in 12, they were placed as a bridge to surgery. In 22 patients (85%), stent placement was successful on the first occasion. In the remaining four individuals, one was successfully stented at the second occasion, and three required emergency surgery. Nine of the 12 patients (75%) in the bridge to-surgery group underwent elective colon resection. In the palliative group, four patients (29%) had reobstruction of the stents, and in one (9%), the stent migrated. In the remaining nine patients (64%), the stent was patent until the patient died or until the time of last follow-up (median, 156 days). CONCLUSIONS: In our experience with 26 patients who developed a complete bowel obstruction as a consequence of a malignant tumor, placement of colonic stents to achieve immediate nonoperative decompression proved to be both safe and effective. Subsequent elective resection was accomplished in the majority of resectable cases. PMID- 12095975 TI - Genetic alterations in normal epithelium of colorectal cancer patients may be a useful indicator for subsequent metachronous tumor development. AB - BACKGROUND: We attempted to identify areas of microsatellite alterations specific to histologically normal colorectal epithelium and to clarify the correlations among those molecular events and clinicopathologic features. METHODS: We conducted a prospective observation study on 51 colorectal cancer patients. Preoperative blood and microdissected histologically normal colorectal epithelium and neoplastic tissues were collected. Microsatellite analyses with seven microsatellite loci were performed to examine the genetic potential of individual tumors and histologically normal colorectal epithelium. RESULTS: In the sporadic colorectal cancer group, p53 LOH in the neoplastic epithelium had a significant correlation with the maximum tumor diameter and the preoperative serum cancer antigen 19-9 level, but not with the depth of invasion of the primary tumor. Among the patients who had p53 LOH in the histologically normal colorectal epithelium, four additional tumors were discovered within 30 months after curative surgery. For those patients, microsatellite alterations in normal colorectal epithelium were more sensitive than tumor markers. CONCLUSIONS: For accurate LOH analysis, nonmalignant lymphocytes from blood should be used as the appropriate normal DNA sample. Focusing on the identification of high-risk patients for microsatellite alterations in histologically normal colorectal epithelium can be a useful indicator of subsequent metachronous tumor development after colorectal surgery. For accurate LOH analysis, nonmalignant lymphocytes from blood should be used as the appropriate normal DNA sample. Focusing on the identification of high-risk patients for microsatellite alterations in histologically normal colorectal epithelium can be a useful indicator of subsequent metachronous tumor development after colorectal surgery. PMID- 12095976 TI - Presence of the human leukocyte antigen class II gene DRB1*1101 predicts interferon gamma levels and disease recurrence in melanoma patients. AB - BACKGROUND: Increased interferon gamma (IFN-gamma) levels are an independent predictor of melanoma recurrence. Human leukocyte antigen (HLA) class II genes can regulate cytokine production; we investigated whether these genes would predict IFN-gamma levels and recurrence in melanoma patients. METHODS: Of 591 patients who presented with localized melanoma, 579 underwent identification of HLA class II alleles; 233 melanoma patients and 90 controls underwent determination of plasma IFN-gamma levels. HLA class II genes were examined for association with IFN-gamma levels and disease recurrence. RESULTS: After a median follow-up of 60 months, melanoma patients with IFN-gamma levels above the mean control value were more likely to have developed disease recurrence compared with patients with levels below the mean. The HLA class II gene HLA-DRB1*1101 was the strongest predictor of recurrence, and HLA-DRB1*1101-positive melanoma patients had increased levels of IFN-gamma compared with patients lacking the gene. CONCLUSIONS: Among patients with localized melanoma, both HLA-DRB1*1101 and increased IFN-gamma levels were associated with an increased risk for recurrence; HLA-DRB1*1101-positive patients had relatively increased levels of IFN-gamma. HLA class II genes may mediate cytokine production in melanoma patients, and this mechanism may help determine the risk of disease recurrence. PMID- 12095977 TI - Radiofrequency ablation of the porcine liver with complete hepatic vascular occlusion. AB - BACKGROUND: We studied the effects of radiofrequency ablation, relative to hepatic blood flow, on the volume and shape of the resulting tissue necrosis. The extent of necrosis is directly proportional to the size of the electrode and inversely related to blood flow, which dissipates the heat generated. METHODS: Two areas of necrosis were created in each of eight porcine livers, which were assigned to four groups according to blood flow occlusion: no occlusion, occlusion of the hepatic artery and portal vein, occlusion of the hepatic veins, and complete hepatic vascular occlusion. After 25 minutes of liver reperfusion, the animals were euthanized, and the livers were examined. RESULTS: Complete vascular occlusion resulted in the greatest area of necrosis (28.6 +/- 3.4 cm(3)), followed by occlusion of the hepatic artery and portal vein (19.2 +/- 5.9 cm(3)), occlusion of hepatic veins (14.4 +/- 2.6 cm(3)), and no occlusion (4.9 +/ 1.5 cm(3)). The volume of the necrotic areas created during complete vascular occlusion were significantly greater than those created with no occlusion, as well as those created with only the hepatic artery and portal vein occluded (P <.05). CONCLUSIONS: Complete vascular occlusion, combined with radiofrequency ablation, increases the volume of necrosis and creates a more spherical ablative area. PMID- 12095979 TI - The expression of the developmentally regulated proto-oncogene Pax-3 is modulated by N-Myc. AB - N-Myc is a member of the Myc family of transcription factors that have been shown to play a pivotal role in cell proliferation and differentiation. In this report, we have investigated the relationship between N-Myc and the developmental control gene Pax-3. Using transient transfection assays, we show that the Pax-3 promoter is activated by both N-Myc-Max and c-Myc-Max. Moreover, we show that Myc regulation of Pax-3 promoter activity is dependent upon a noncanonical E box site in the 5' promoter region of Pax-3. In addition, we show that ectopic expression of both N-Myc and c-Myc leads to increased expression of Pax-3 mRNA. Furthermore, we show that Pax-3 mRNA expression is cell cycle-regulated and that the 5' promoter region of Pax-3 (bp -1578 to +56) can direct cell cycle-dependent gene expression with kinetics similar to that of the endogenous transcript. Site directed mutagenesis of the E box site within the Pax-3 promoter significantly altered the pattern of expression through the cell cycle. These results suggest that the Myc family of transcription factors may modulate Pax-3 expression in vivo. PMID- 12095978 TI - Intratumoral dihydropyrimidine dehydrogenase messenger RNA level reflects tumor progression in human colorectal cancer. AB - BACKGROUND: Determination of intratumoral dihydropyrimidine dehydrogenase (DPD) is of clinical interest because increased DPD levels can influence the tumor response to 5-fluorouracil-based chemotherapy through increased inactivation of the agent in tumor cells. METHODS: DPD messenger RNA (mRNA) levels were evaluated in 80 consecutive patients undergoing surgery for primary colorectal cancer and 12 cases of liver metastasis. RESULTS: Higher DPD mRNA levels were associated with higher pathologic classification, corresponding to the T categories (r =.267; P =.003). The DPD mRNA level was statistically higher in tumors with microscopic lymph node metastasis than in those without (P =.002). Hence, the DPD mRNA level increased in accordance with Dukes' classification (r =.387; P =.0001). The DPD mRNA level of the liver metastasis from colorectal cancer was significantly higher than that of primary lesions (P =.002). In eight patients, the DPD mRNA level of the liver metastasis was significantly higher than that of the matched primary tumor (P =.017). CONCLUSIONS: Increases of the DPD mRNA level in cancerous tissue seem to reflect tumor progression. High DPD mRNA levels in liver metastasis and advanced colorectal cancer may have clinical importance for 5-fluorouracil-based chemosensitivity. PMID- 12095980 TI - Signal transduction from N-cadherin increases Bcl-2. Regulation of the phosphatidylinositol 3-kinase/Akt pathway by homophilic adhesion and actin cytoskeletal organization. AB - Associated with the metastatic progression of epithelial tumors is the dynamic regulation of cadherins. Whereas E-cadherin is expressed in most epithelium and carcinomas, recent studies suggest that the up-regulation of other cadherin subtypes in carcinomas, such as N-cadherin, may function in cancer progression. We demonstrate that a signal transduction cascade links the N-cadherin.catenin adhesion complex to up-regulation of the anti-apoptotic protein Bcl-2. In suspension, aggregates of DU-145 cells, an E-cadherin expressing human prostate carcinoma line, survive loss of integrin-dependent adhesion by a different anti apoptotic signaling pathway than the N-cadherin expressing lines PC3 and PC3N. N cadherin intercellular adhesion mediates a 3.5-fold increase in Bcl-2 protein expression, whereas the level of the proapoptotic protein Bax remains constant. Only N-cadherin ligation in PC3 cells, which express both N-cadherin and E cadherin, is sufficient to induce activation of Akt/protein kinase B. N-cadherin homophilic ligation initiates phosphatidylinositol 3-kinase-dependent activation of Akt resulting in Akt phosphorylation of Bad on serine 136. Following N cadherin homophilic adhesion phosphatidylinositol 3-kinase was identified in immunoprecipitates of the N-cadherin.catenin complex. The recruitment of phosphatidylinositol 3-kinase to the adhesion complex is dependent on ligation of N-cadherin and an organized actin cytoskeleton because cytochalasin D blocks the recruitment. We propose that N-cadherin homophilic adhesion can initiate anti apoptotic signaling, which enhances the Akt cell survival pathway in metastatic cancer. PMID- 12095981 TI - Transcellular proteolysis demonstrated by novel cell surface-associated substrates of dipeptidyl peptidase IV (CD26). AB - Proteolytic enzymes contribute to the regulation of cellular functions such as cell proliferation and death, cytokine production, and matrix remodeling. Dipeptidyl peptidase IV (DP IV) catalyzes the cleavage of several cytokines and thereby contributes to the regulation of cytokine production and the proliferation of immune cells. Here we show for the first time that cell surface bound DP IV catalyzes the cleavage of specific substrates that are associated with the cellular surface of neighboring cells. Rhodamine 110 (R110), a highly fluorescent xanthene dye, was used to synthesize dipeptidyl peptidase IV (DP IV/CD26) substrates Gly(Ala)-Pro-R110-R, thus facilitating a stable binding of the fluorescent moiety on the cell surface. The fixation resulted from the interaction with the reactive anchor rhodamine and allowed the quantification of cellular DP IV activity on single cells. The reactivity, length, and hydrophobicity of rhodamine was characterized as the decisive factor that facilitated the determination of cellular DP IV activity. Using fluorescence microscopy, it was possible to differentiate between different DP IV activities. The hydrolysis of cell-bound substrates Xaa-Pro-R110-R by DP IV of neighboring cells and by soluble DP IV was shown using flow cytometry. These data demonstrate that ectopeptidases such as DP IV may be involved in communication between blood cells via proteolysis of cell-associated substrates. PMID- 12095982 TI - Three adjacent serines in the extracellular domains of the CaR are required for L amino acid-mediated potentiation of receptor function. AB - The extracellular calcium (Ca(2+)(o))-sensing receptor (CaR) is a key player in Ca(2+)(o) homeostasis. The activity of CaR can be potentiated by various l-amino acids. In this study, we examined whether conserved amino acid residues involved in the binding of glutamate to metabotropic glutamate receptors (mGluRs) also participate in the potentiation of the activity of CaR by l-phenylalanine. Ser 170 corresponding to Thr-188 in rat mGluR1a appears to be important for the modulating actions of phenylalanine. In the presence of phenylalanine, a mutant CaR with a single mutation S170A showed no significant decrease in its EC(50) for stimulation by Ca(2+)(o) and a modest increase in its maximal activity. In addition, mutating Ser-169 and Ser-171 together with Ser-170 yielded a more complete block of the phenylalanine modulation than did the single mutation. The presence of the triple mutation, S169A/S170A/S171A, also eliminated phenylalanine potentiation of the activities of heterodimeric receptors in which one of the monomeric receptors had intact triple serines (A877Stop). The putative amino acid binding site of the CaR is probably close to or structurally dependent on the Ca(2+)(o) binding sites of the receptor, because mutant CaRs with mutations in the putative amino acid binding site exhibited severely reduced responses to Ca(2+)(o). PMID- 12095983 TI - Protein kinase C(alpha) is required for vanilloid receptor 1 activation. Evidence for multiple signaling pathways. AB - Activation of vanilloid receptor (VR1) by protein kinase C (PKC) was investigated in cells ectopically expressing VR1 and primary cultures of dorsal root ganglion neurons. Submicromolar phorbol 12,13-dibutyrate (PDBu), which stimulates PKC, acutely activated Ca(2+) uptake in VR1-expressing cells at pH 5.5, but not at mildly acidic or neutral pH. PDBu was antagonized by bisindolylmaleimide, a PKC inhibitor, and ruthenium red, a VR1 ionophore blocker, but not capsazepine, a vanilloid antagonist indicating that catalytic activity of PKC is required for PDBu activation of VR1 ion conductance, and is independent of the vanilloid site. Chronic PDBu dramatically down-regulated PKC(alpha) in dorsal root ganglion neurons or the VR1 cell lines, whereas only partially influencing PKCbeta, delta, -epsilon, and -zeta. Loss of PKC(alpha) correlated with loss of response to acute re-challenge with PDBu. Anandamide, a VR1 agonist in acidic conditions, acts additively with PDBu and remains effective after chronic PKC down regulation. Thus, two independent VR1 activation pathways can be discriminated: (i) direct ligand binding (anandamide, vanilloids) or (ii) extracellular ligands coupled to PKC by intracellular signaling. Experiments in cell lines co expressing VR1 with different sets of PKC isozymes showed that acute PDBu-induced activation requires PKC(alpha), but not PKC(epsilon). These studies suggest that PKC(alpha) in sensory neurons may elicit or enhance pain during inflammation or ischemia. PMID- 12095984 TI - Sanglifehrin A acts as a potent inhibitor of the mitochondrial permeability transition and reperfusion injury of the heart by binding to cyclophilin-D at a different site from cyclosporin A. AB - Cyclosporin A (CsA) inhibits opening of the mitochondrial permeability transition pore (MPTP), a critical event in some forms of necrotic and apoptotic cell death, by binding to cyclophilin D (CyP-D) and inhibiting its peptidyl-prolyl cis-trans isomerase (PPIase) activity. Sanglifehrin A (SfA), like CsA, exerts its immunosuppressive action by binding to cyclophilin A but at a different site from CsA, and unlike the latter, SfA does not inhibit calcineurin activity. Here we demonstrate that SfA inhibits the PPIase activity of CyP-D (K(0.5) 2 nm) and acts as a potent inhibitor of MPTP opening under both energized and de-energized conditions. However, unlike CsA, the dose-response curve for inhibition by SfA is sigmoidal rather than hyperbolic, suggesting a multimeric structure for the MPTP with cooperativity between subunits. Furthermore, SfA does not prevent CyP-D binding to submitochondrial particles or detergent-solubilized adenine nucleotide translocase (ANT), implying that CyP-D binding to the ANT does not require PPIase activity but pore opening does. Once bound to the MPTP, SfA is not readily dissociated, and inhibition of pore opening is maintained following extensive washing. To investigate the potential of SfA as an inhibitor of cell death in vivo, we used the Langendorff perfused rat heart. SfA caused a time-dependent inhibition of the MPTP that was maintained on mitochondrial isolation to a greater extent than was CsA inhibition. We demonstrate that SfA, like CsA, improves the recovery of left ventricular developed pressure during reperfusion after 30 min of global ischemia and greatly reduces lactate dehydrogenase release, implying inhibition of necrotic damage. Because SfA does not inhibit calcineurin activity, our data suggest that it may be more desirable than CsA for protecting tissues recovering from ischemic episodes and for studying the role of the MPTP in cell death. PMID- 12095985 TI - Molecular cloning and characterization of STAMP1, a highly prostate-specific six transmembrane protein that is overexpressed in prostate cancer. AB - We have identified a novel gene, six transmembrane protein of prostate 1 (STAMP1), which is largely specific to prostate for expression and is predicted to code for a 490-amino acid six transmembrane protein. Using a form of STAMP1 labeled with green fluorescent protein in quantitative time-lapse and immunofluorescence confocal microscopy, we show that STAMP1 is localized to the Golgi complex, predominantly to the trans-Golgi network, and to the plasma membrane. STAMP1 also localizes to vesicular tubular structures in the cytosol and colocalizes with the early endosome antigen 1 (EEA1), suggesting that it may be involved in the secretory/endocytic pathways. STAMP1 is highly expressed in the androgen-sensitive, androgen receptor-positive prostate cancer cell line LNCaP, but not in androgen receptor-negative prostate cancer cell lines PC-3 and DU145. Furthermore, STAMP1 expression is significantly lower in the androgen dependent human prostate xenograft CWR22 compared with the relapsed derivative CWR22R, suggesting that its expression may be deregulated during prostate cancer progression. Consistent with this notion, in situ analysis of human prostate cancer specimens indicated that STAMP1 is expressed exclusively in the epithelial cells of the prostate and its expression is significantly increased in prostate tumors compared with normal glands. Taken together, these data suggest that STAMP1 may have an important role in the normal prostate cell as well as in prostate cancer progression. PMID- 12095986 TI - Identification of mammalian mitochondrial translational initiation factor 3 and examination of its role in initiation complex formation with natural mRNAs. AB - Human mitochondrial translational initiation factor 3 (IF3(mt)) has been identified from the human expressed sequence tag data base. Using consensus sequences derived from conserved regions of the bacterial IF3, several partially sequenced cDNA clones were identified, and the complete sequence was assembled in silico from overlapping clones. IF3(mt) is 278 amino acid residues in length. MitoProt II predicts a 97% probability that this protein will be localized in mitochondria and further predicts that the mature protein will be 247 residues in length. The cDNA for the predicted mature form of IF3(mt) was cloned, and the protein was expressed in Escherichia coli in a His-tagged form. The mature form of IF3(mt) has short extensions on the N and C termini surrounding a region homologous to bacterial IF3. The region of IF3(mt) homologous to prokaryotic factors ranges between 21-26% identical to the bacterial proteins. Purified IF3(mt) promotes initiation complex formation on mitochondrial 55 S ribosomes in the presence of mitochondrial initiation factor 2 (IF2(mt)), [(35)S]fMet-tRNA, and either poly(A,U,G) or an in vitro transcript of the cytochrome oxidase subunit II gene as mRNA. IF3(mt) shifts the equilibrium between the 55 S mitochondrial ribosome and its subunits toward subunit dissociation. In addition, the ability of E. coli initiation factor 1 to stimulate initiation complex formation on E. coli 70 S and mitochondrial 55 S ribosomes was investigated in the presence of IF2(mt) and IF3(mt). PMID- 12095988 TI - Role of ubiquilin associated with protein-disulfide isomerase in the endoplasmic reticulum in stress-induced apoptotic cell death. AB - Up-regulation of several stress proteins such as heat-shock proteins and glucose regulated proteins participate in tolerance against environmental stress. Previously, we found that protein-disulfide isomerase (PDI) is specifically up regulated in response to hypoxia/brain ischemia in astrocytes. In addition, the overexpression of this gene into neurons protects against apoptotic cell death induced by hypoxia/brain ischemia. To address the detailed function of PDI, we screened for proteins that interact with PDI using the yeast two-hybrid system. We report here that PDI interacts with ubiquilin, which has a ubiquitin-like domain and a ubiquitin-associated domain. Interestingly, ubiquilin is also up regulated in response to hypoxia in glial cells with a time course similar to that of PDI induction. In hypoxia-treated glial cells, the endogenous ubiquilin and PDI were almost completely co-localized, suggesting that ubiquilin is an endoplasmic reticulum-associated protein. Overexpression of this gene in neuronal cells resulted in significant inhibition of the DNA fragmentation triggered by hypoxia, but not that induced by nitric oxide or staurosporine. Moreover, ubiquilin has the ability to attenuate CHOP induction by hypoxia. These observations suggested that ubiquilin together with PDI have critical functions as regulatory proteins for CHOP-mediated cell death, and therefore up-regulation of these proteins may result in acquisition of tolerance against ischemic stress in glial cells. PMID- 12095987 TI - Fibroblast growth factor 1 regulates signaling via the glycogen synthase kinase 3beta pathway. Implications for neuroprotection. AB - We hypothesize that in neurodegenerative disorders such as Alzheimer's disease and human immunodeficiency virus encephalitis the neuroprotective activity of fibroblast growth factor 1 (FGF1) against several neurotoxic agents might involve regulation of glycogen synthase kinase-3beta (GSK3beta), a pathway important in determining cell fate. In primary rat neuronal and HT22 cells, FGF1 promoted a time-dependent inactivation of GSK3beta by phosphorylation at serine 9. Blocking FGF1 receptors with heparinase reduced this effect. The effects of FGF1 on GSK3beta were dependent on phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) because inhibitors of this pathway or infection with dominant negative Akt adenovirus blocked inactivation. Furthermore, treatment of neuronal cells with FGF1 resulted in ERK-independent Akt phosphorylation and beta-catenin translocation into the nucleus. On the other hand, infection with wild-type GSK3beta recombinant adenovirus-associated virus increased activity of GSK3beta and cell death, both of which were reduced by FGF1 treatment. Moreover, FGF1 protection against glutamate toxicity was dependent on GSK3beta inactivation by the PI3K-Akt but was independent of ERK. Taken together these results suggest that neuroprotective effects of FGF1 might involve inactivation of GSK3beta by a pathway involving activation of the PI3K-Akt cascades. PMID- 12095989 TI - A bipartite Ca2+-regulated nucleoside-diphosphate kinase system within the Chlamydomonas flagellum. The regulatory subunit p72. AB - Regulation of flagellar activity in Chlamydomonas involves both Ca(2+) and cAMP mediated signaling pathways. However, Chlamydomonas and sea urchin sperm flagella also exhibit nucleoside-diphosphate kinase (NDK) activity, suggesting a requirement for GTP within this highly conserved organelle. In sea urchin sperm, the NDK catalytic subunit is an integral component of the outer dynein arm. Here we describe a modular protein (p72) from the Chlamydomonas flagellum that consists of three domains closely related to the presumptive regulatory segment of rat NDK-7 followed by two EF-hands that are predicted to bind Ca(2+). There are close homologues of p72 in both mammalian and insect genomes. The p72 protein is tightly associated with the flagellar axoneme and is located along the entire length except at the transition zone. Cross-linking experiments suggest that p72 interacts with two or three additional axonemal polypeptides. The sensitivity of p72 to tryptic digestion differed considerably in the presence and the absence of Ca(2+), suggesting that it indeed binds this ligand. These studies indicate that the flagellar NDK system is bipartite with the regulatory and catalytic components residing on different polypeptides. We propose that Ca(2+) regulation of flagellar motility in Chlamydomonas may be achieved in part through a downstream GTP-mediated signaling pathway. PMID- 12095990 TI - Fatty acid infusion selectively impairs insulin action on Akt1 and protein kinase C lambda /zeta but not on glycogen synthase kinase-3. AB - To determine the mechanism(s) for insulin resistance induced by fatty acids, we measured the ability of insulin to activate phosphoinositide 3-kinase (PI3K) and multiple distal pathways in rats. Following a 5-h infusion of lipid or glycerol (control), rats underwent a euglycemic hyperinsulinemic clamp. Insulin stimulated IRS-1-associated PI3K activity in muscle of glycerol-infused rats 2.4-fold but had no effect in lipid-infused rats. IRS-2- and phosphotyrosine-associated PI3K activity were increased 3.5- and 4.8-fold, respectively, by insulin in glycerol infused rats but only 1.6- and 2.3-fold in lipid-infused rats. Insulin increased Akt1 activity 3.9-fold in glycerol-infused rats, and this was impaired 41% in lipid-infused rats. Insulin action on Akt2 and p70S6K were not impaired, whereas activation of protein kinase C lambda/zeta activity was reduced 47%. Insulin inhibited glycogen synthase kinase 3alpha (GSK-3alpha) activity by 30% and GSK 3beta activity by approximately 65% and increased protein phosphatase-1 activity by 40-47% in both glycerol- and lipid-infused rats. Insulin stimulated glycogen synthase activity 2.0-fold in glycerol-infused rats but only 1.4-fold in lipid infused rats. Thus, 1) elevation of fatty acids differentially affects insulin action on pathways distal to PI3K, impairing activation of Akt1 and protein kinase C lambda/zeta and 2) insulin action on glycogen synthase can be regulated independent of effects on GSK-3 and protein phosphatase-1 activity in vivo. PMID- 12095991 TI - Naringin-sensitive phosphorylation of plectin, a cytoskeletal cross-linking protein, in isolated rat hepatocytes. AB - To identify phosphoproteins that might play a role in naringin-sensitive hepatocellular cytoskeletal disruption and apoptosis induced by algal toxins, hepatocyte extracts were separated by gel electrophoresis and immunostained with a phosphothreonine-directed antibody. Use of dilute (5%) polyacrylamide gels containing 6 m urea allowed the resolution of one very large (approximately 500 kDa) okadaic acid- and naringin-sensitive phosphoprotein, identified by tryptic fingerprinting, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and immunostaining as the cytolinker protein, plectin. The naringin sensitive phosphorylation induced by okadaic acid and microcystin-LR probably reflected inhibition of a type 2A protein phosphatase, whereas the naringin resistant phosphorylation induced by calyculin A, tautomycin, and cantharidin probably involved a type 1 phosphatase. Okadaic acid caused a collapse of the plectin-immunostaining bile canalicular sheaths and the general cytoskeletal plectin network into numerous medium-sized plectin aggregates. Inhibitors of protein kinase C, cAMP-dependent protein kinase, or Ca(2+)/calmodulin-dependent kinase II had moderate or no protective effects on plectin network disruption, whereas naringin offered 86% protection. Okadaic acid induced a naringin sensitive phosphorylation of AMP-activated protein kinase (AMPK), the stress activated protein kinases SEK1 and JNK, and S6 kinase. The AMPK-activating kinase (AMPKK) is likely to be the target of inhibition by naringin, the other kinases serving as downstream components of an AMPKK-initiated signaling pathway. PMID- 12095992 TI - Mutation analysis of Lhca1 antenna complex. Low energy absorption forms originate from pigment-pigment interactions. AB - The light harvesting complex Lhca1, one of the four gene products comprising the photosystem I antenna system, has been analyzed by site-directed mutagenesis with the aim of determining the chromophore(s) responsible for its long wavelength chlorophyll spectral form, a specific characteristic of the LHCI antenna complex. A family of mutant proteins, each carrying a mutation at a single chlorophyll binding residue, was obtained and characterized by biochemical and spectroscopic methods. A map of the chromophores bound to each of the 10 chlorophyll-binding sites was drawn, and the energy levels of the Q(y) transition were determined in most cases. When compared with Lhcb proteins previously analyzed, Lhca1 is characterized by stronger interactions between individual chromophores as detected by both biochemical and spectroscopic methods; most mutations, although targeted to a single residue, lead to the loss of more than one chromophore and of conservative CD signals typical of chlorophyll-chlorophyll interactions. The lower energy absorption form (686 nm at 100K, 688 nm at room temperature), which is responsible for the red-shifted emission components at 690 and 701 nm, typical of Lhca1, is associated with a chlorophyll a/chlorophyll a excitonic interaction originating from a pigment cluster localized in the protein domain situated between helix C and the helix A/helix B cross. This cluster includes chlorophylls bound to sites A5-B5-B6 and a xanthophyll bound to site L2. PMID- 12095993 TI - HIV-1 Vpr induces apoptosis through caspase 9 in T cells and peripheral blood mononuclear cells. AB - Human immunodeficiency virus, type 1 (HIV-1), vpr gene encodes a 14-kDa virion associated protein, which exhibits significant effects on human cells. One important property of Vpr is its ability to induce apoptosis during infection. Apoptotic induction is likely to play a role in the pathogenesis of AIDS. However, the pathway of apoptosis is not clearly defined. In this report we investigate the mechanism of apoptosis induced by HIV-1 Vpr using a Vpr pseudotype viral infection system or adeno delivery of Vpr in primary human lymphoid cells and T-cells. With either vector, HIV-1 Vpr induced cell cycle arrest at the G(2)/M phase and apoptosis in lymphoid target cells. Furthermore, we observed that with both vectors, caspase 9, but not caspase 8, was activated following infection of human peripheral blood mononuclear cell with either Vpr positive HIV virions or adeno-delivered Vpr. Activation of the caspase 9 pathway resulted in caspase 3 activation and apoptosis in human primary cells. These effects were coincident with the disruption of the mitochondrial transmembrane potential and induction of cytochrome c release by Vpr. The Vpr-induced signaling pathway did not induce CD95 or CD95L expression. Bcl-2 overexpressing cells succumb to Vpr-induced apoptosis. These studies illustrate that Vpr induces a mitochondria-dependent apoptotic pathway that is distinct from apoptosis driven by the Fas-FasL pathway. PMID- 12095994 TI - A thyroid hormone antagonist that inhibits thyroid hormone action in vivo. AB - We have characterized the newly developed thyroid hormone antagonist NH-3 in both cell culture and in vivo model systems. NH-3 binds Xenopus laevis thyroid hormone receptors directly in vitro and induces a conformation distinct from agonist bound receptors. Transcriptional activation of a thyroid hormone response element containing reporter gene is strongly inhibited by NH-3 in a dose-dependent manner. In addition, NH-3 prevents X. laevis thyroid hormone receptors from binding to the p160 family of co-activators GRIP-1 and SRC-1 in a two-hybrid assay. To assess the potency of the compound in vivo, we used induced and spontaneous X. laevis tadpole metamorphosis, a thyroid hormone-dependent developmental process. NH-3 inhibits thyroid hormone-induced morphological changes in a dose-dependent manner and inhibits the up-regulation of endogenous thyroid hormone-responsive genes. Spontaneous metamorphosis is efficiently and reversibly arrested by NH-3 with at least the same effectiveness as the thyroid hormone synthesis inhibitor methimazole. Therefore, NH-3 is the first thyroid hormone antagonist to demonstrate potent inhibition of thyroid hormone action in both cell culture- and whole animal-based assays. PMID- 12095995 TI - Mitochondrial substrate level phosphorylation is essential for growth of procyclic Trypanosoma brucei. AB - Oxidative phosphorylation and substrate level phosphorylation catalyzed by succinyl-CoA synthetase found in the citric acid and the acetate:succinate CoA transferase/succinyl-CoA synthetase cycle contribute to mitochondrial ATP synthesis in procyclic Trypanosoma brucei. The latter pathway is specific for trypanosome but also found in hydrogenosomes. In organello ATP production was studied in wild-type and in RNA interference cell lines ablated for key enzymes of each of the three pathways. The following results were obtained: 1) ATP production in the acetate:succinate CoA transferase/succinyl-CoA synthetase cycle was directly demonstrated. 2) Succinate dehydrogenase appears to be the only entry point for electrons of mitochondrial substrates into the respiratory chain; however, its activity could be ablated without causing a growth phenotype. 3) Growth of procyclic T. brucei was not affected by the absence of either a functional citric acid or the acetate:succinate CoA transferase/succinyl-CoA synthetase cycle. However, interruption of both pathways in the same cell line resulted in a growth arrest. In summary, these results show that oxygen independent substrate level phosphorylation either linked to the citric acid cycle or tied into acetate production is essential for growth of procyclic T. brucei, a situation that may reflect an adaptation to the partially hypoxic conditions in the insect host. PMID- 12095996 TI - Pigment binding of photosystem I light-harvesting proteins. AB - Light-harvesting complexes (LHC) of higher plants are composed of at least 10 different proteins. Despite their pronounced amino acid sequence homology, the LHC of photosystem II show differences in pigment binding that are interpreted in terms of partly different functions. By contrast, there is only scarce knowledge about the pigment composition of LHC of photosystem I, and consequently no concept of potentially different functions of the various LHCI exists. For better insight into this issue, we isolated native LHCI-730 and LHCI-680. Pigment analyses revealed that LHCI-730 binds more chlorophyll and violaxanthin than LHCI 680. For the first time all LHCI complexes are now available in their recombinant form; their analysis allowed further dissection of pigment binding by individual LHCI proteins and analysis of pigment requirements for LHCI formation. By these different approaches a correlation between the requirement of a single chlorophyll species for LHC formation and the chlorophyll a/b ratio of LHCs could be detected, and indications regarding occupation of carotenoid-binding sites were obtained. Additionally the reconstitution approach allowed assignment of spectral features observed in native LHCI-680 to its components Lhca2 and Lhca3. It is suggested that excitation energy migrates from chlorophyll(s) fluorescing at 680 (Lhca3) via those fluorescing at 686/702 nm (Lhca2) or 720 nm (Lhca3) to the photosystem I core chlorophylls. PMID- 12095997 TI - Amyloid fibril formation by pentapeptide and tetrapeptide fragments of human calcitonin. AB - The process of amyloid fibril formation by the human calcitonin hormone is associated with medullary thyroid carcinoma. Based on the effect of pH on the fibrillization of human calcitonin, the analysis of conformationally constrained analogues of the hormone, and our suggestion regarding the role of aromatic residues in the process of amyloid fibril formation, we studied the ability of a short aromatic charged peptide fragment of calcitonin (NH(2)-DFNKF-COOH) to form amyloid fibrils. Here, using structural and biophysical analysis, we clearly demonstrate the ability of this short peptide to form well ordered amyloid fibrils. A shorter truncated tetrapeptide, NH(2)-DFNK-COOH, also formed fibrils albeit less ordered than those formed by the pentapeptide. We could not detect amyloid fibril formation by the NH(2)-FNKF-COOH tetrapeptide, the NH(2)-DFN-COOH tripeptide, or the NH(2)-DANKA-COOH phenylalanine to the alanine analogue of the pentapeptide. The formation of amyloid fibrils by rather hydrophilic peptides is quite striking, because it was speculated that hydrophobic interactions might play a key role in amyloid formation. This is the first reported case of fibril formation by a peptide as short as a tetrapeptide and one of very few cases of amyloid formation by pentapeptides. Because the aromatic nature seems to be the only common property of the various very short amyloid-forming peptides, it further supports our hypothesis on the role of aromatic interactions in the process of amyloid fibril formation. PMID- 12095998 TI - Combinatorial control of yeast FET4 gene expression by iron, zinc, and oxygen. AB - Acquisition of metals such as iron, copper, and zinc by the yeast Saccharomyces cerevisiae is tightly regulated. High affinity uptake systems are induced under metal-limiting conditions to maintain an adequate supply of these essential nutrients. Low affinity uptake systems function when their substrates are in greater supply. The FET4 gene encodes a low affinity iron and copper uptake transporter. FET4 expression is regulated by several environmental factors. In this report, we describe the molecular mechanisms underlying this regulation. First, we found that FET4 expression is induced in iron-limited cells by the Aft1 iron-responsive transcriptional activator. Second, FET4 is regulated by zinc status via the Zap1 transcription factor. We present evidence that FET4 is a physiologically relevant zinc transporter and this provides a rationale for its regulation by Zap1. Finally, FET4 expression is regulated in response to oxygen by the Rox1 repressor. Rox1 attenuates activation by Aft1 and Zap1 in aerobic cells. Derepression of FET4 may allow the Fet4 transporter to play an even greater role in metal acquisition under anaerobic conditions. Thus, Fet4 is a multisubstrate metal ion transporter under combinatorial control by iron, zinc, and oxygen. PMID- 12095999 TI - Antimicrobial resistance and susceptibility testing: an evergreen topic. PMID- 12096000 TI - Bicyclic pyrimidine nucleoside analogues (BCNAs) as highly selective and potent inhibitors of varicella-zoster virus replication. AB - Bicyclic pyrimidine nucleoside analogues (BCNAs) represent highly potent and selective inhibitors of varicella-zoster virus (VZV) replication in cell culture. The compounds inhibit a variety of clinical VZV strains, in the higher picomolar range, whilst being non-toxic at micromolar concentrations. The compounds do not inhibit the closely related simian varicella virus or any other viruses, including herpes simplex virus type 1 (HSV-1), HSV-2 and cytomegalovirus. The BCNAs owe at least part of their antiviral selectivity to a specific activation/phosphorylation by the VZV-encoded thymidine kinase (TK) and associated thymidylate kinase (dTMP-K) activity, while being not recognized by the closely related HSV-1-encoded TK/dTMP-K enzyme. In addition, the 5' monophosphates of BCNAs are neither a substrate nor an inhibitor of the cellular dTMP-K, and are not subject of back-conversion to the corresponding nucleosides by 5'-deoxynucleotidases. In contrast to the anti-HSV-1/VZV drug (E)-5-(2 bromovinyl)-2'-deoxyuridine (BVDU), the BCNAs are not catabolized by human (erythrocyte) or bacterial (Escherichia coli) thymidine phosphorylase to release the free bicyclic pyrimidine base. Also, unlike BVU (the free base of BVDU), the BCNA bases do not inhibit dihydropyrimidine dehydrogenase. Consequently, the catabolism of the anticancer drug 5-fluorouracil (5-FU) is not influenced by the BCNA base in cell-free enzyme assays or in mice that were exposed to combinations of 5-FU with BCNAs or their free base. BCNAs have a good oral bioavailability and, owing to their highly lipophilic nature, are assumed to be able to cross the blood-brain barrier efficiently. Given the above-mentioned favourable properties, BCNAs may represent a promising novel class of highly selective anti-VZV drugs that should be further pursued for clinical application. PMID- 12096001 TI - Identification of PSE and OXA beta-lactamase genes in Pseudomonas aeruginosa using PCR-restriction fragment length polymorphism. AB - OBJECTIVE: A method using PCR-restriction fragment length polymorphism was developed to identify Pseudomonas aeruginosa beta-lactamase genes. METHODS: Two hundred and fifty-nine P. aeruginosa isolates were screened by PCR with 11 primer pairs designed to detect genes encoding PSE, OXA, TEM and SHV enzymes. PSE and OXA gene variants were distinguished by restriction of PCR products with endonucleases recognizing sites involved in point mutations. Nucleotide sequences were verified for a few isolates by sequencing the PCR products. RESULTS: Four isolates produced extended-spectrum beta-lactamases (ESBLs) according to the double disc synergy test. PCR detecting bla(PSE) genes was positive in 162 (62.5%) isolates: 151 carried bla(PSE-1) and 11 carried a variant encoding an enzyme differing from PSE-1 by a single amino acid substitution (Pro102 to Ser). PCR detecting sequences for enzymes of the OXA-10 group was positive in 68 (26.3%) isolates: 31 carried bla(OXA-10), one carried bla(OXA-14) and 36 carried a new variant intermediate between bla(OXA-13) and bla(OXA-19). The bla(OXA-2) gene was identified in 13 (5%) isolates. Two other isolates carried bla(OXA-2) variants encoding ESBLs differing from OXA-2 by a single amino acid substitution (Asp150 to Tyr and Trp159 to Cys, respectively). PCR detecting sequences for enzymes of the OXA-1 group was positive in 12 (4.6%) isolates. A bla(TEM) gene was identified in five (1.9%) isolates (three bla(TEM-1), one bla(TEM-2), one bla(TEM-4)). CONCLUSION: This approach is effective for screening P. aeruginosa for beta-lactamase gene carriage in epidemiological studies and for detecting new variants. PMID- 12096002 TI - Effect of growth phase and pH on the in vitro activity of a new glycopeptide, oritavancin (LY333328), against Staphylococcus aureus and Enterococcus faecium. AB - Oritavancin (LY333328) is a novel glycopeptide with activity against Gram positive bacteria such as methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecium. We compared the effects of pH and growth phase on the activity of oritavancin and vancomycin against methicillin resistant S. aureus and vancomycin-susceptible and -resistant E. faecium. Killing curve methods were used to evaluate the effect of growth phase (stationary versus exponential) and pH (6.4, 7.4 and 8.0). An inoculum of 10(6) cfu/mL was used for all experiments. Growth phase of S. aureus and vancomycin-susceptible E. faecium did not influence the rate and killing activity of oritavancin. The rate of killing by oritavancin against the vancomycin-resistant E. faecium strain was significantly faster and the reduction in cfu/mL at 24 h was significantly greater when the organism was in exponential compared with stationary growth phase (P < 0.05). In exponential growth phase, time to 99.9% killing was achieved in 0.6 +/- 0.01 h for the vancomycin-resistant strain, whereas in stationary growth phase, oritavancin did not decrease the inoculum by 99.9% within 24 h. Oritavancin's activity against S. aureus and vancomycin-susceptible E. faecium was not influenced by the pH conditions tested. Oritivancin's killing activity against the vancomycin-resistant E. faecium strain was significantly enhanced when tested at pH 7.4 and 8.0 (P < 0.05). Our study has demonstrated that oritavancin's activity does not seem to be influenced by the growth phase of the organisms or the pH of the environment when tested against sensitive strains of S. aureus and E. faecium. However, oritavancin's activity might be reduced against vancomycin-resistant E. faecium strains in stationary growth phase, as seen in infective endocarditis or when organisms are exposed to an acidic environment. PMID- 12096003 TI - Two novel assays for the detection of haemin-binding properties of antimalarials evaluated with compounds isolated from medicinal plants. AB - Forty-two compounds isolated from nine plants used within South America for the treatment of malaria were tested for haemin binding using two novel, rapid screening methods. The data obtained were analysed with respect to IC(50) values for in vitro toxicity to Plasmodium falciparum trophozoites. One method, a multiwell assay based on the inhibition of the interaction of haemin with glutathione (GSH), is sensitive in the 10 microM range, takes c. 1 h and is suitable for either a high throughput screen or rapid assay during natural product isolation. Of 19 compounds showing antiplasmodial activity (IC(50) < 40 microM), 16 (84%) showed >40% inhibition of GSH-haemin reaction. The sensitivity and specificity of the assay were 0.85 and 0.82, respectively. The positive predictive value was 0.81 and the negative predictive value 0.86. A more sensitive assay (0.1 microM range) is based on the reversal by haemin-binding compounds of the haemin inhibition of the L-dopachrome-methyl ester tautomerase activity of human macrophage migration inhibitory factor. This assay gives a better idea of the affinity of interaction and uses very small amounts of test compound. The log[RI(50)] of eight of the compounds that tested positive in the above assays together with those of quinine and chloroquine showed a positive correlation with log[antiplasmodial IC(50)] for strain T9-96 (r = 0.824) and strain K1 (r = 0.904). Several of the antimalarial compounds that bind haemin are isoquinolines, a class not shown previously to interact with haemin. PMID- 12096004 TI - Activity of cefditoren against respiratory pathogens. AB - The activity of cefditoren and six other cephalosporins was tested against 250 pneumococci, including strains resistant to macrolides and quinolones. Cefditoren gave the lowest MICs, with MIC(50) and MIC(90) values of < or =0.016/0.03, 0.125/0.5 and 0.5/2.0 mg/L for penicillin-susceptible, -intermediate and resistant pneumococci, respectively. A time-kill study of 12 pneumococcal strains with varying drug susceptibilities showed that cefditoren, at 2 x MIC, gave 99% killing of all strains after 12 h, with 99.9% killing after 24 h. Other cephalosporins gave similar kill kinetics but at higher concentrations. Against 160 Haemophilus influenzae, cefditoren had the lowest MICs (MIC(50) and MIC(90) both < or =0.016 mg/L), irrespective of beta-lactamase production. Time-kill studies of cefditoren compared with five other oral cephalosporins showed that cefditoren, at 8 x MIC, was bactericidal against 8/9 strains and gave 90% killing of all strains at the MIC after 12 h. Activity was bactericidal (99.9% killing) after 24 h with all drugs tested. Multistep studies of four penicillin susceptible, four penicillin-intermediate and four penicillin-resistant strains showed that cefditoren, co-amoxiclav and cefprozil did not select for resistant mutants after 50 subcultures, compared with cefuroxime and azithromycin, where resistant mutants were selected in two and nine strains, respectively. Single step mutation studies showed that cefditoren, at the MIC, had the lowest frequency of spontaneous mutants compared with other drugs. PMID- 12096005 TI - Streptococcus pneumoniae bacteraemia in Belgium: differential characteristics in children and the elderly population and implications for vaccine use. AB - The characteristics of bacteraemia with Streptococcus pneumoniae in children (0-4 years) and the elderly (> or =60 years) were compared over a 7 year period (1994 2000). Of a total of 7927 isolates of invasive S. pneumoniae studied in the national reference laboratory, 74% (n = 5837) were blood isolates. Of these 5837 S. pneumoniae bacteraemias, 843 (14%) occurred in children and 3144 (54%) in the elderly. The prevalence of penicillin resistance (MIC > or = 0.1 mg/L) in bacteraemic isolates rose from 8.2% to 18.9% (P = 0.03) in children and from 5.1% to 16.35% (P = 0.001) in the elderly over the study period. The prevalence of erythromycin resistance (MIC > or= 1 mg/L) in bacteraemic isolates was significantly higher in children than in the elderly (44.7% versus 25.7%, P = 0.001) and rose significantly over the 7 year period in the elderly (18.6-33.65%, P = 0.001). There were more serogroups and serotypes (SGTs) among the bacteraemic isolates obtained from the elderly compared with children (36 versus 26, P = 0.03). SGTs 6, 14, 18 and 19 cause significantly more bacteraemia in children than in the elderly. The opposite is true for SGTs 3, 7, 8, 9, 11, 12, 15, 20, 22 and 35. The new 7, 9 and 11 valent conjugate vaccine formulations cover significantly more bacteraemic SGTs in children than in the elderly (82%, 89.5% and 92% versus 55.5%, 65% and 77.5%, respectively; P = 0.001). The 23 valent polysaccharide vaccine provides a theoretical coverage of 95% in the elderly population. Our data indicate consideration of a vaccination strategy in the elderly population that combines the efficacy of conjugate vaccines with the broad coverage of the 23 valent polysaccharide vaccine. PMID- 12096006 TI - Effect of ciprofloxacin on lethal and sublethal challenge with endotoxin and on early cytokine responses in a murine in vivo model. AB - The influence of ciprofloxacin on immune responses has been suggested by results of in vitro and in vivo studies. This effect was studied using a murine model that measured mortality and early cytokine responses after challenge with endotoxin. C57/BL6 mice weighing between 18 and 21 g were given a single intraperitoneal dose of lipopolysaccharide (LPS), ranging from 200 to 1000 microg. Mice were pre-treated with an intraperitoneal injection of 5% dextrose in sterile water containing 0.0-6.0 mg of ciprofloxacin 1 h before LPS challenge. Cytokine responses were assessed by measuring concentrations in serum separated from blood obtained by cardiac puncture of anaesthetized mice at 0, 1, 3, 6 and 24 h following LPS administration. Mice were observed for 72 h following administration of LPS and serum cytokines were measured using ELISA. More than 4.5 mg of ciprofloxacin (675-900 mg/m(2) or 225-300 mg/kg) given 1 h before LPS challenge consistently protected mice from a lethal dose of LPS (14/14 versus 0/7, P < 0.00001). Ciprofloxacin significantly attenuated the production of tumour necrosis factor-alpha and interleukin-12 response after LPS challenge. In addition, ciprofloxacin significantly increased serum interleukin-10 concentrations but had little or no effect on interleukin-6 or interleukin-1beta serum concentrations. Similar effects were evident with sublethal doses of LPS and were most pronounced at the lowest dose of LPS studied. These observations indicate that ciprofloxacin can prevent endotoxin-mediated death and alter early host cytokine responses. This effect may influence the course of infection in a manner that is independent of the drug's antimicrobial activity. PMID- 12096007 TI - Macrolide-treated Pseudomonas aeruginosa induces paradoxical host responses in the lungs of mice and a high mortality rate. AB - OBJECTIVE: Accumulating data have demonstrated that macrolide antibiotics suppress Pseudomonas aeruginosa virulence, which may explain the efficacy of macrolides in clinical settings. We examined the virulence of macrolide-treated bacteria in vivo. METHODS: P. aeruginosa PAO-1 was grown for 24 h on agar containing sub-MIC antibiotics, and then mice were challenged intranasally with 10(7) cfu of bacteria. RESULTS AND CONCLUSIONS: The mortality rate of mice inoculated with bacteria grown in the presence of clarithromycin (10 mg/L), erythromycin (10 mg/L) or azithromycin (5 mg/L) was 80%, 80% and 100%, respectively. In contrast, none of the mice inoculated with non-treated bacteria or bacteria treated with other antibiotics died. Lung weight and protein concentration in bronchoalveolar lavage fluid (BALF) were significantly higher in the clarithromycin group between 3 and 9 h. Moreover, we detected higher levels of tumour necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) in the BALF of these mice. These data demonstrate that macrolide-treated P. aeruginosa induced paradoxically strong responses, such as elevation of TNF-alpha, NO and permeability in the lungs. PMID- 12096008 TI - Pharmacokinetic/pharmacodynamic modelling of ciprofloxacin 250 mg/12 h versus 500 mg/24 h for urinary infections. AB - A simulation study was performed to evaluate and compare the standard dosage regimen of 250 mg/12 h versus 500 mg/24 h of ciprofloxacin for the treatment of urinary tract infections (UTIs). Pharmacokinetic parameters reported for healthy young and old individuals were used for the simulation of drug levels in urine, at different mean urine flow rates (1-2.5 L/day). Pharmacokinetic/pharmacodynamic analysis of the results revealed that 500 mg ciprofloxacin once a day produced a more favourable profile in urine than 250 mg/12 h, particularly in the elderly, due to the slower elimination of the drug in this group of patients. Circadian rhythms were also considered for the simulation of drug levels in urine. According to the results, 500 mg once a day administered in the morning would be a better choice than 250 mg/12 h at least for uncomplicated UTI; nevertheless, clinical assays are needed to prove this hypothesis. PMID- 12096009 TI - Penetration of linezolid into bone, fat, muscle and haematoma of patients undergoing routine hip replacement. AB - Twelve patients undergoing total hip replacement were given 600 mg of linezolid as a 20 min iv infusion along with conventional prophylaxis of 1 g of cefamandole immediately before surgery. Routine total hip arthroplasty was carried out, and at timed intervals during surgery samples of bone, fat, muscle and blood were collected for assay by high-performance liquid chromatography analysis. Samples of the haematoma fluid that formed around the operation site and further blood samples for assay were also collected at timed intervals following the operation. The penetration of linezolid into bone was rapid, with mean concentrations of 9.1 mg/L (95% CI 7.7-10.6 mg/L) achieved at 10 min after the infusion, decreasing to 6.3 mg/L (95% CI 3.9-8.6 mg/L) at 30 min. Correction for the simultaneous blood concentrations gave mean values for bone penetration of 51% at 10 min, 60% at 20 min and 47% at 30 min. Although the penetration of linezolid into fat was also rapid, mean concentrations and degree of penetration were c. 60% of those in bone; at 10 min they were 4.5 mg/L (95% CI 3.0-6.1 mg/L; penetration 27%); at 20 min they were 5.2 mg/L (95% CI 4.0-6.4 mg/L; penetration 37%); and at 30 min, 4.1 mg/L (95% CI 3.3-4.8 mg/L; penetration 31%). For muscle the corresponding values were 10.4 mg/L (95% CI 8.1-12.7 mg/L; penetration 58%) at 10 min, 13.4 mg/L (95% CI 10.2-16.5 mg/L; penetration 94%) at 20 min and 12.0 mg/L (95% CI 9.2-14.8 mg/L; penetration 93%) at 30 min. Mean concentrations of linezolid in the haematoma fluid drained from around the operation site were 8.2 mg/L at 6-8 h and 5.6 mg/L at 10-12 h after the infusion, and 7.0 mg/L at 2-4 h following a second 600 mg infusion given 12 h post-operatively. We conclude that linezolid exhibits rapid penetration into bone, fat and muscle of patients undergoing hip arthroplasty, to achieve levels in excess of its MIC for susceptible organisms (< or=4 mg/L); therapeutic concentrations were maintained in the haematoma fluid that surrounds the operation site for >16 h. PMID- 12096010 TI - Cefepime plus amikacin versus piperacillin-tazobactam plus amikacin for initial antibiotic therapy in haematology patients with febrile neutropenia: results of an open, randomized, multicentre trial. AB - BACKGROUND: Standard therapy for suspected infections in patients with profound neutropenia is the combination of a beta-lactam antibiotic plus an aminoglycoside. Cefepime's broad-spectrum activity makes it an option for initial empirical therapy in neutropenic patients. The aim of this study is to evaluate the efficacy and safety of cefepime plus amikacin compared with piperacillin tazobactam plus amikacin for initial empirical treatment of fever in adult haematology patients with severe neutropenia. METHODS: In this prospective multicentre trial, 969 patients with 984 febrile neutropenic episodes were randomized to receive iv amikacin (20 mg/kg every 24 h) combined with either cefepime (2 g every 8 h) or piperacillin-tazobactam (4 g/500 mg every 6 h). Clinical response was determined at 72 h and at completion of therapy. RESULTS: Eight hundred and sixty-seven episodes were assessable for efficacy (432 cefepime, 435 piperacillin-tazobactam). The frequency of success without modification of the empirical therapy was nearly identical for cefepime plus amikacin (49%) compared with piperacillin-tazobactam plus amikacin (51%). Similar rates of success were found for microbiologically documented infection: 40% versus 39%, respectively. Antibiotic modification was necessary in 49% of cefepime and 44% of piperacillin-tazobactam patients. The overall response rate, with or without modification of the assigned treatment, was 94% in both groups. Drug-related adverse events were reported in 10% of cefepime plus amikacin versus 11% of piperacillin-tazobactam plus amikacin patients. Mortality due to infection occurred in a total of 10 patients (two cefepime, eight piperacillin-tazobactam). CONCLUSION: The empirical regimen of cefepime plus amikacin is equivalent to piperacillin-tazobactam plus amikacin in febrile adult haematology patients with severe neutropenia. KEYWORDS: cefepime, piperacillin-tazobactam, amikacin, empirical antibiotic therapy, febrile neutropenia, haematological malignancy PMID- 12096011 TI - In vitro activity of a novel oxazolidinone, AZD2563, against randomly selected and multiresistant Gram-positive cocci. AB - The in vitro activity of AZD2563, a novel oxazolidinone, was assessed against 595 Gram-positive cocci, comprising recent surveillance isolates and a collection of resistant (including multiresistant), epidemiologically diverse isolates. The MICs of AZD2563 for staphylococci, pneumococci and enterococci had narrow ranges, 0.25-2 mg/L, with modal MICs of 1 mg/L for staphylococci and pneumococci, and 1-2 mg/L for enterococci. AZD2563 was equally active against the surveillance isolates and those that had been selected for their multiresistance to other agents. The MICs of AZD2563 were either the same as those of linezolid or two fold lower. PMID- 12096012 TI - Antipneumococcal activity of AZD2563, a new oxazolidinone, compared with nine other agents. AB - The in vitro activity of AZD2563, a new oxazolidinone, was compared with that of linezolid, vancomycin, quinupristin/dalfopristin, amoxicillin, levofloxacin, penicillin, erythromycin, azithromycin and clindamycin against a range of pneumococci by microdilution and time-kill studies. Against 300 pneumococci (99 penicillin susceptible, 86 penicillin intermediate, 115 penicillin resistant, 185 erythromycin resistant, 35 quinolone resistant), both oxazolidinones remained active against isolates less susceptible to other agents, with MICs ranging between 0.125 and 2 mg/L; AZD2563 MICs were generally one dilution lower than those of linezolid. Both quinupristin/dalfopristin and vancomycin were active against all groups (MIC ranges 0.125-2 and 0.125-0.25 mg/L, respectively). Apart from 35 isolates with levofloxacin MICs > or= 8 mg/L, levofloxacin MICs were < or =0.25-4 mg/L. MICs of amoxicillin and erythromycin rose with penicillin G MICs; most macrolide-resistant isolates were either penicillin-intermediate or resistant. Against 16 organisms with differing beta-lactam, macrolide and quinolone MICs, time-kill studies showed that AZD2563 was bactericidal (99.9% killing) at 4 x MIC against nine strains at 24 h, with 90% killing of all 16 strains at 2 x MIC after 12 h. Similar results were obtained with linezolid. Both oxazolidinones were bacteriostatic at the MIC against all 16 strains. Amoxicillin, levofloxacin and vancomycin, at 2 x MIC, were bactericidal against 15 of the 16 strains after 24 h. Quinupristin/dalfopristin yielded the most rapid killing, with bactericidal activity against 13 of 16 strains at the MIC after 3 h and against 15 strains at 2 x MIC after 24 h. Erythromycin was bactericidal against all 10 strains with MICs < or= 8 mg/L at 4 x MIC after 24 h. PMID- 12096013 TI - Anti-anaerobic activity of AZD2563, a new oxazolidinone, compared with eight other agents. AB - The anti-anaerobic activity of AZD2563, a new oxazolidinone, was compared with that of eight other agents against 201 Gram-positive and 99 Gram-negative anaerobes. MIC(50) and MIC(90) values (mg/L) for the 201 Gram-positive organisms were as follows: AZD2563, 1.0/4.0; linezolid, 1.0/4.0; quinupristin/dalfopristin, 0.5/1.0; amoxicillin, 0.25/1.0; clindamycin, 0.25/8.0; metronidazole, 0.5/>16.0; vancomycin, 0.5/2.0; teicoplanin, 0.125/0.25; and meropenem, 0.06/1.0. AZD2563, linezolid, vancomycin, teicoplanin and quinupristin/dalfopristin were mainly active against Gram-positive anaerobes, with no useful activity against Gram negative anaerobes. PMID- 12096014 TI - Assay of teicoplanin in serum: comparison of high-performance liquid chromatography and fluorescence polarization immunoassay. AB - A simple liquid extraction coupled with reverse-phase HPLC and UV detection was shown to correlate well with fluorescence polarization immunoassay (FPIA) on the Abbott TD(x) analyser for serum teicoplanin analysis, r(2) = 0.974, HPLC = 0.908 TDx + 2.324. A Bland-Altman plot showed no significant bias between the results. The HPLC method was linear over the range 10-100 mg/L. The HPLC method showed very good reproducibility, comparable to FPIA. The inter-assay coefficients of variation were 2.76%, 2.29% and 2.65% at 10.3, 51.4 and 77.1 mg/L, respectively, with intra-assay coefficients of variation of 1.86%, 1.97% and 1.17% at 9.13, 50.65 and 75.1 mg/L, respectively. Recoveries were between 99.1 and 101.8% within the analytical range. The HPLC method described is simple, robust, highly reproducible and suited to a clinical laboratory with the appropriate equipment. PMID- 12096015 TI - 2-Alkoxycarbonylaminopyridines: inhibitors of Mycobacterium tuberculosis FtsZ. AB - Compounds originally designed as putative tubulin inhibitors were tested as antitubercular agents for inhibition of the Mycobacterium tuberculosis analogue of tubulin, FtsZ. Initial screening of 200 2-alkoxycarbonylpyridines found several that inhibited M. tuberculosis growth. Two compounds, SRI-3072 and SRI 7614, inhibited FtsZ polymerization and were equipotent against susceptible and single-drug-resistant strains of M. tuberculosis. In addition, SRI-3072 reduced the growth of M. tuberculosis in mouse bone marrow macrophages. Our results suggest that these types of compound might be developed into antitubercular drugs effective against the current multidrug-resistant strains of M. tuberculosis. PMID- 12096016 TI - Susceptibility testing of Aspergillus spp. by means of an automated blood culture system. AB - The BacT/Alert blood culture system was evaluated for the MIC determination of amphotericin B for 27 strains of Aspergillus from five different species. An inoculum of c. 10(8) cfu/mL and an incubation time of 48 h resulted in MICs for 26 of 27 strains that corresponded with published MICs obtained in a microdilution assay. Twelve of 13 strains with MIC(48) values >/=2 mg/L showed growth within 24 h in bottles containing 0.25 mg/L amphotericin B, indicating that results can be obtained the day following inoculation. It is concluded that the BacT/Alert blood culture system can be used for susceptibility testing of Aspergillus spp. with amphotericin B. PMID- 12096017 TI - In vitro activity of three new triazoles and one echinocandin against Candida bloodstream isolates from cancer patients. AB - The in vitro activities of voriconazole, posaconazole, ravuconazole and micafungin were compared with those of fluconazole, itraconazole, ketoconazole, flucytosine and amphotericin B against 164 candidaemia isolates recovered from cancer patients in two Canadian centres. The MIC(50) results for ravuconazole, voriconazole, posaconazole and micafungin were 0.01, 0.03, 0.12 and 0.25 mg/L, respectively. The new antifungal agents showed substantial activity against isolates demonstrating in vitro resistance to fluconazole and itraconazole. These results suggest that the newer antifungal agents possess promising activity against invasive Candida isolates, particularly against those with reduced susceptibility to fluconazole and itraconazole. PMID- 12096018 TI - Penicillin tolerance amongst non-toxigenic Corynebacterium diphtheriae isolated from cases of pharyngitis. AB - Twenty-four strains of non-toxigenic Corynebacterium diphtheriae biotype gravis from the throats of patients with pharyngitis/tonsillitis were assayed for susceptibility to penicillin and erythromycin using determination of MIC, MBC and time-kill curves. There were no differences between the MICs of penicillin for susceptible and tolerant strains. All but one strain had penicillin MBCs > or = 2 mg/L. Seventy-one per cent (17/24) of the strains were tolerant to penicillin. In contrast, all strains were susceptible to erythromycin (MIC < or = 0.016 mg/L). These aspects should be considered when choosing the therapy for treating non toxigenic C. diphtheriae pharyngitis/tonsillitis. PMID- 12096019 TI - In vitro anti-Helicobacter pylori activity of BAS-118, a new benzamide derivative. AB - The antibacterial activity of BAS-118, a new benzamide derivative, against Helicobacter pylori and other species of bacteria was investigated, as was the in vitro ability of the compound to induce drug resistance in H. pylori. The MICs of BAS-118 for 155 isolates of H. pylori, including 30 clarithromycin (CAM) resistant isolates (MIC > or= 1.56 mg/L) and 25 metronidazole (MNDZ)-resistant isolates (MIC >or = 6.25 mg/L), and 29 reference strains of other genera were determined by an agar dilution method. The MIC(50), MIC(90) and MIC range of BAS 118 for 100 randomly selected isolates of H. pylori were or =8 mg/L. In summary, BAS 118 is a novel anti-H. pylori agent with a potent and selective antibacterial activity, which includes CAM- and MNDZ-resistant isolates. Furthermore, BAS-118 does not appear to induce drug resistance readily in vitro. PMID- 12096020 TI - Effect of plaunotol in combination with clarithromycin or amoxicillin on Helicobacter pylori in vitro and in vivo. AB - Plaunotol, a cytoprotective anti-ulcer agent, has antibacterial activity against Helicobacter pylori. The purpose of the present study was to investigate the effect of plaunotol when combined with clarithromycin or amoxicillin against H. pylori. When combined with clarithromycin, plaunotol showed synergic activity against 11 of 14 strains, and additive activity against the other three strains, by chequerboard titration. When combined with amoxicillin, plaunotol showed additive activity against 10 of 14 strains. No antagonistic effects were seen against any of the strains tested. The interactions between plaunotol and either clarithromycin or amoxicillin were determined by time-kill assay against the Sydney Strain (strain SS1) of H. pylori. The combination of plaunotol with clarithromycin showed synergic activity and with amoxicillin showed additive activity. In a C57BL/6 mouse gastritis model infected with H. pylori SS1, the plaunotol-clarithromycin and plaunotol-amoxicillin combinations both exhibited synergic effects, which allowed the effective dose of clarithromycin to be reduced when co-administered with plaunotol. These results suggest that plaunotol may have a useful role in combination with anti-H. pylori drugs in the treatment of H. pylori-associated diseases. PMID- 12096021 TI - Biocide abuse and antimicrobial resistance: being clear about the issues. PMID- 12096023 TI - In vitro activity of a novel des-fluoro(6) quinolone, garenoxacin (BMS-284756), against rapidly growing mycobacteria and Nocardia isolates. PMID- 12096024 TI - Group B Streptococcus highly resistant to gentamicin. PMID- 12096025 TI - Reduction of MIC of azithromycin for Salmonella typhi and Staphylococcus aureus in broth exposed to previous growth of Salmonella typhi. PMID- 12096026 TI - In vitro activities of the ketolides ABT-773 and telithromycin and of three macrolides against genetically characterized isolates of Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae and Moraxella catarrhalis. PMID- 12096027 TI - Pivotal role of CCL25 (TECK)-CCR9 in the formation of gut cryptopatches and consequent appearance of intestinal intraepithelial T lymphocytes. AB - Cryptopatches (CP) are murine gut anatomical sites for generating thymus independent intraepithelial T lymphocytes (IEL). However, it remains elusive how lympho-hematopoietic progenitor cells migrate from bone marrow (BM) into CP and differentiate into IEL. Here we show that mice reconstituted with BM-derived c kit(+) cells express CCL25 (TECK)-intrakine gene, which reduces specifically the chemotactic response to CCL25 but not CXCL12 in the thymocytes. These mice exhibited a dramatic reduction of CP and IEL in the small intestine, and harbored conspicuously decreased numbers of c-kit(+) cells in the emaciated CP. In contrast, T cells in the thymic, splenic and lymph node compartments developed normally in these mice. Importantly, it was demonstrated that CD11c(+) dendritic stromal cells in CP expressed CCL25 and c-kit(+) Lin(-) BM cells displayed vigorous chemotactic response to CCL25. Furthermore, RT-PCR analysis detects mRNA expression of CCR9 in the c-kit(+) Lin(-) BM cells. Thus, these results demonstrate that the CCL25-CCR9 pathway is essential for CP formation and the consequent appearance of IEL. PMID- 12096028 TI - Endotoxin can induce MyD88-deficient dendritic cells to support T(h)2 cell differentiation. AB - Toll-like receptor (TLR) signaling activates dendritic cells (DC) to secrete proinflammatory cytokines and up-regulate co-stimulatory molecule expression, thereby linking innate and adaptive immunity. A TLR-associated adapter protein, MyD88, is essential for cytokine production induced by TLR. However, in response to a TLR4 ligand, lipopolysaccharide (LPS), MyD88-deficient (MyD88(-/-)) DC can up-regulate co-stimulatory molecule expression and enhance their T cell stimulatory activity, indicating that the MyD88-independent pathway through TLR4 can induce some features of DC maturation. In this study, we have further characterized function of LPS-stimulated, MyD88(-/-) DC. In response to LPS, wild type DC could enhance their ability to induce IFN-gamma production in allogeneic mixed lymphocyte reaction (alloMLR). In contrast, in response to LPS, MyD88(-/-) DC augmented their ability to induce IL-4 instead of IFN-gamma in alloMLR. Impaired production of T(h)1-inducing cytokines in MyD88(-/-) DC cannot fully account for their increased T(h)2 cell-supporting ability, because absence of T(h)1-inducing cytokines in DC caused impairment of IFN-gamma, but did not lead to augmentation of IL-4 production in alloMLR. In vivo experiments with adjuvants also revealed T(h)2-skewed immune responses in MyD88(-/-) mice. These results demonstrate that the MyD88-independent pathway through TLR4 can confer on DC the ability to support T(h)2 immune responses. PMID- 12096029 TI - Role of CD28 co-stimulation in generation and maintenance of virus-specific T cells. AB - Efficient induction of T cell responses is normally assumed to require both TCR mediated signaling and engagement of co-stimulatory molecules, in particular CD28. However, the importance of CD28 co-stimulation in induction and maintenance of antiviral T cell responses is not clearly established. For this reason antiviral CD4(+) and CD8(+) T cell responses in CD28-deficient mice were studied using two different viruses [vesicular stomatitis virus and lymphocytic choriomeningitis virus (LCMV)]. Intracellular cytokine staining and/or MHC peptide tetramers were used to enumerate antigen-specific T cells. In addition, we used DNA constructs encoding viral epitopes to probe the importance of the epitope itself. Our results reveal that while the context of antigen presentation (live virus versus DNA construct) is a critical factor in determining the requirement for CD28 co-stimulation, epitope and virus dose play little if any role. Direct visualization of antigen-specific cells also confirms the notion that CD28 is more critical for the generation of antiviral T(h)1 cells than for T(c)1 cells generated in response to the same virus (LCMV). Most importantly, the present study reveals that CD28 generally is essential for the host to respond optimally over a broad set of conditions, and our results may imply that the relatively CD28 independent activation of LCMV-specific CD8(+) T cells may represent an extreme situation related to the non-cytolytic nature of this virus allowing the delivery of a uniquely strong and prolonged signal 1. PMID- 12096030 TI - Exosomes bearing HLA-DR1 molecules need dendritic cells to efficiently stimulate specific T cells. AB - Exosomes are small vesicles (60-100 nm) secreted by various cell types upon the fusion of endosomal compartments with the plasma membrane. Exosomes from antigen presenting cells (APC), such as B lymphocytes and dendritic cells (DC), bear MHC class II molecules. In addition, the injection of DC-derived exosomes was reported to elicit potent T cell responses in vivo. Here, we analyzed the activation of specific T cells by MHC class II-bearing exosomes in vitro. The rat mast cell line, RBL-2H3, was engineered to express human class II molecules uniformly loaded with an antigenic peptide [HLA-DR1-hemagglutinin (HA)]. These cells secreted exosomes bearing DR1 class II molecules upon stimulation by a calcium ionophore or IgE receptor cross-linking. Exosomes bearing DR1-HA(306-318) complexes activated HA/DR1-specific T cells only weakly, whereas the cross linking of such exosomes to latex beads increased stimulation of specific T cells. By contrast, the incubation of free exosomes with DC resulted in the highly efficient stimulation of specific T cells. Thus, exosomes bearing MHC class II complexes must be taken up by professional APC for efficient T cell activation. PMID- 12096031 TI - Identification of three genes up-regulated in PU.1 rescued monocytic precursor cells. AB - The requirement of the transcription factor PU.1 for macrophage development has been well documented. However, the target genes regulated by PU.1 controlling macrophage maturation are not known. A granulocyte macrophage colony stimulating factor (GM-CSF)-dependent PU.1 null monocytic precursor cell was stably transduced with a PU.1-expressing retrovirus. The expression of PU.1 altered the surface expression of a few proteins expressed on monocytes; these cells, however, remained GM-CSF dependent and maintained an immature phenotype. In contrast to the PU.1 null cells, the cells expressing PU.1 responded to macrophage colony stimulating factor (M-CSF) with subsequent development into mature macrophages. Using suppressive subtractive hybridization between the PU.1 null and immature PU.1 rescued cells, three genes, MRP-14, Dap12 and CD53, were found expressed in the rescued cells, but not in the PU.1 null cells. In addition, these genes were modulated during M-CSF-induced maturation of the PU.1 rescued cells. The PU.1 null and rescued early monocytic cells provide a useful model to study the role of PU.1 in macrophage development. PMID- 12096032 TI - Molecular mechanisms of lipopolysaccharide-induced cyclooxygenase-2 expression in human neutrophils: involvement of the mitogen-activated protein kinase pathway and regulation by anti-inflammatory cytokines. AB - Neutrophils are an important cellular source of proinflammatory mediators, whose regulation may be of potential benefit for the treatment of a number of inflammatory diseases. However, the mechanisms of lipopolysaccharide (LPS) induced neutrophil activation and its regulation by anti-inflammatory cytokines have not yet been fully elucidated. Recent studies have revealed that mitogen activated protein kinases (MAPK) play a crucial role in the generation of proinflammatory mediators in some cell types. Therefore, we conducted this study to determine whether MAPK activation could be involved in prostaglandin E(2) (PGE(2)) production and cyclooxygenase (COX)-2 expression in LPS-stimulated human neutrophils. PD98059 (MEK1 inhibitor) and SB203580 (p38(MAPK) inhibitor) reduced PGE(2) production as well as COX-2 expression in LPS-stimulated neutrophils. In addition, both extracellular signal-regulated protein kinase (ERK) and p38(MAPK) were phosphorylated and activated in time- and dose-dependent manners. Since we previously showed that IL-10 and IL-4 similarly inhibited COX-2 expression in LPS stimulated neutrophils, we next tested the effects of IL-10 and IL-4 on the phosphorylation and activation of both kinases. IL-10 inhibited the phosphorylation and activation of p38(MAPK), but not ERK. In addition, IL-4 caused a marginal inhibition in the activation of p38(MAPK). Taken together, these results suggest that both ERK and p38(MAPK) pathways are involved in LPS induced COX-2 expression and PGE(2) production in neutrophils, and IL-10 and IL-4 inhibit neutrophil prostanoid synthesis by down-regulating the activation of p38(MAPK). PMID- 12096033 TI - Generation of potent T(h)1 responses from patients with lymphoid malignancies after differentiation of B lymphocytes into dendritic-like cells. AB - Dendritic cells (DC) are a group of potent antigen-presenting cells (APC) specialized for initiating T cell immune responses. They originate from the bone marrow and upon stimulation with bacterial products, cytokines or CD40 ligation they acquire the ability to migrate to the secondary lymphoid organs. In vitro DC can be generated from human CD34(+) bone marrow cells and CD14(+) peripheral blood monocytes after culture with different cytokine combinations. Since most leukemic cells and tumors in general are devoid of APC capacities, various strategies have been used to increase their recognition and confer the capacity of antigen presentation on them. Because of our interest in the design of vaccine immunotherapy protocols for the adjuvant treatment of patients with lymphoid malignancies (LM), we chose to explore the capacity of human acute lymphoblastic leukemia, chronic lymphocytic leukemia and plasma cell leukemia to differentiate into cells with APC and DC features. Our results among a sample of 10 patients demonstrate that such approach is feasible. Leukemic cells could be induced in the presence of IL-4 and CD40L to exhibit a DC morphology with a phenotype of mature DC-like cells. They could also induce a potent proliferative response in naive CD4(+) T cells. In addition, they expressed chemokine receptor CCR7 and CD62L, and could drive T cells towards a T(h)1 response with secretion of IFN gamma. Our strategy leading to increased LM cell immunogenicity may have potential clinical applications and LM appear to be attracting candidates for adjuvant vaccination and adoptive immunotherapy. PMID- 12096034 TI - Differential role of E-selectin and P-selectin in T lymphocyte migration to cutaneous inflammatory reactions induced by cytokines. AB - E-selectin and P-selectin are thought to be important in the infiltration of T lymphocytes in inflammation, but their role in cytokine-induced cutaneous inflammatory reactions has not been examined. A technique for quantifying labeled T lymphocyte migration to cytokine-induced dermal inflammation in mice was developed. After i.v. injection, (51)Cr-labeled T lymphocytes migrated to lesions induced by IFN-gamma and tumor necrosis factor (TNF)-alpha, and in even greater numbers to the combination of IFN-gamma + TNF-alpha, and to sites injected with concanavalin A (Con A). In E-selectin mAb-treated and in E-selectin-deficient mice, IFN-gamma-, IFN-gamma + TNF-alpha- and Con A-induced T cell accumulation was inhibited by 45-65%, but TNF-alpha-induced infiltration was unaffected. In P selectin mAb-treated and P-selectin-deficient mice, T cell accumulation remained unchanged in most of the lesions. Combined, E-selectin and P-selectin mAb treatment inhibited T cell accumulation in all four types of reactions, and significantly more than E-selectin blockade alone in migration to Con A. Results in E-selectin- and P-selectin-deficient mice confirmed these observations, and demonstrated strain-dependent differences in the contributions of the two selectins. In conclusion, T cells migrating to dermal inflammatory reactions utilize both E-selectin and P-selectin, but alternate adhesion pathways also contribute, since blocking both endothelial selectins does not abolish T cell migration. P-selectin plays a less important role than E-selectin, since blocking E-selectin, but not P-selectin, alone decreased T cell accumulation. The relative contribution of the selectins varies depending on the initiating inflammatory stimulus and the genetic background. PMID- 12096035 TI - In vivo induction of tolerance by an Ig peptide is not affected by the deletion of FcR or a mutated IgG Fc fragment. AB - To induce tolerance to a variety of epitopes, we have designed a gene therapy approach in which peptides or antigens are expressed in frame on a soluble IgG fusion protein scaffold and delivered via retroviral gene therapy in B cells in vivo. Initially, tolerance to the lambda repressor cI sequence p1-102 or its immunodominant epitopes (e.g. p12-26 or p73-88) was elicited in both T cells and B cells when lipopolysaccharide (LPS) blasts are transduced and injected into naive or even primed recipients. While a role of secreted Ig fusion protein in this process is not clear, we have previously demonstrated the importance of antigen presentation on MHC class II of B cell antigen-presenting cells (APC) for tolerance induction. To further examine the role of the Ig and especially of the Fc portion of the IgG in tolerogenesis, we transduced LPS blasts from FcR gamma II(-/-), Fc gamma RI(-/-), Fc gamma RIII(-/-), FcR(-/-) or naive mice with retroviral vectors expressing IgG1-102, Delta IgG1-102 (mutated construct on position 297 of the Fc portion) or IgG12-26. When these transduced LPS blasts from FcR knockout mice were injected into normal (or knockout) syngeneic recipient mice, they induced tolerance both to the immunodominant epitopes and the full-length protein in that the antibody responses to the immunodominant epitopes were reduced. In this paper, we show that this tolerance resides at both the T and B cell level. Moreover, mutation of residue 297, which affects IgG functions including FcR binding, did not alter the tolerogenicity of the construct. These results suggest that the Fc portion of the IgG molecules is not required for humoral nor for cellular tolerance induction using the IgG-antigen tolerogens. PMID- 12096036 TI - IFN-gamma-inducible expression of thymus and activation-regulated chemokine/CCL17 and macrophage-derived chemokine/CCL22 in epidermal keratinocytes and their roles in atopic dermatitis. AB - Thymus and activation-regulated chemokine (TARC)/CCL17 and macrophage-derived chemokine (MDC)/CCL22 are a pair of CC chemokines known to selectively attract T(h)2 type memory T cells via CCR4. Here we examined circulating levels of TARC and MDC in patients with atopic dermatitis (AD) and control subjects by using plasma samples, which reflect blood contents of chemokines more accurately than serum samples. The plasma levels of TARC and MDC were significantly elevated in AD patients. These values also strongly correlated with disease severity and serum lactate dehydrogenase levels, and weakly correlated with serum total IgE levels and blood eosinophilia. Previous studies demonstrated TARC immunoreactivity in the epidermal layer of AD lesional skin and production of TARC by a human keratinocytic cell line HaCaT upon stimulation with IFN-gamma. Here we demonstrated MDC immunoreactivity in the epidermal layer of AD skin at levels stronger than that of TARC. Furthermore, primary epidermal keratinocytes expressed both TARC and MDC mRNA upon stimulation with IFN-gamma, but efficiently secreted only MDC. These results suggest a post-transcriptional regulation in TARC production. IFN-gamma also induced TARC and MDC mRNA in mouse skin. Collectively, both TARC and MDC play important roles in the local accumulation of T(h)2 cells in AD lesional skin. Production of T(h)2-attracting chemokines by epidermal keratinocytes upon treatment with IFN-gamma, which is also the potent inducer of T(h)1-attracting chemokines, may underline the pivotal role of IFN gamma in the chronic phase of AD where both T(h)1 and T(h)2 responses are mixed. PMID- 12096037 TI - The assembly of functional beta(2)-microglobulin-free MHC class I molecules that interact with peptides and CD8(+) T lymphocytes. AB - Functional MHC class I molecules are expressed on the cell surface in the absence of beta(2)-microglobulin (beta(2)m) light chain that can interact with CD8(+) T lymphocytes. Whether their assembly requires peptide binding and whether their recognition by CD8(+) T lymphocytes involves the presentation of peptide epitopes remains unknown. We show that beta(2)m-free H-2D(b) assembles with short peptides that are approximately 9 amino acid residues in length, akin to ligands associated with completely assembled beta(2)m(+) H-2D(b). Remarkably, a subset of the peptides associated with the beta(2)m-free H-2D(b) has an altered anchor motif. However, they also include peptides that contain a beta(2)m(+)H-2D(b) binding anchor motif. Further, the H-2K(b)- and H-2D(b)-restricted peptide epitopes derived from SV-40 T antigen also assemble with H-2(b) class I in beta(2)m-deficient cells and are recognized by epitope-specific CD8(+) T lymphocytes. Taken together our data reveal that functional MHC class I molecules assemble in the absence of beta(2)m with peptides and form CD8(+) T lymphocyte epitopes. PMID- 12096038 TI - A variety of microbial components induce tolerance to lipopolysaccharide by differentially affecting MyD88-dependent and -independent pathways. AB - Exposure of macrophages to lipopolysaccharide (LPS) induces a hypo-responsive state to a second challenge with LPS that is termed LPS tolerance. LPS tolerance is also induced by pre-exposure to lipopeptides and lipoteichoic acid, which trigger Toll-like receptor (TLR) 2-mediated signaling. LPS signaling involves at least two pathways: a MyD88-dependent cascade that is essential for production of inflammatory cytokines and a MyD88-independent cascade that mediates the expression of IFN-inducible genes. We analyzed the induction of LPS tolerance by several microbial components in mouse peritoneal macrophages. Pre-exposure to LPS led to impaired activation of both the pathways. In contrast, mycoplasmal lipopeptides did not affect the MyD88-independent pathway, but impaired the MyD88 dependent signaling by inhibiting LPS-mediated activation of IL-1 receptor associated kinase (IRAK) 1. The induction of LPS tolerance by recently identified TLR ligands was analyzed. Pretreatment with double-stranded RNA, which triggers the activation of TLR3, led to defective activation of the MyD88-independent, but not the MyD88-dependent, pathway. Imidazoquinoline compounds, which are recognized by TLR7, had no effect on the MyD88-independent pathway, but inhibited LPS-induced activation of MyD88-dependent signaling through down-regulation of IRAK1 expression. Thus, each microbial component induced LPS tolerance in macrophages. PMID- 12096039 TI - IL-2-secreting recombinant bacillus Calmette Guerin can overcome a Type 2 immune response and corticosteroid-induced immunosuppression to elicit a Type 1 immune response. AB - The efficacy of bacillus Calmette Guerin (BCG) as a vaccine against tuberculosis is adversely affected by both genetic and environmental factors on the immune system. In this study we have demonstrated that a recombinant BCG (rBCG) secreting biologically active IL-2 has the ability to induce a T(h)1 profile in both immunocompromised and in IL-4 transgenic (Tg) mice. Dexamethasone (DXM) was administered orally to mice prior to vaccination with either rBCG or normal BCG (nBCG). Six weeks post-vaccination with rBCG, splenocytes from DXM-treated mice exhibited a strong antigen-specific proliferative response, while also secreting large amounts of IFN-gamma and low levels of IgG1. The opposite profile occurred when DXM-treated mice were vaccinated with nBCG. Splenocytes from these mice showed no significant proliferation and produced a cytokine profile associated with a T(h)2 immune response, in addition to exhibiting high levels of serum IgG1. In the IL-4 Tg model, mice vaccinated with rBCG again produced a strong T(h)1 immune response, exhibiting a high antigen-specific IFN-gamma:IL-4 ratio and a concomitantly high IgG2a:IgG1 ratio. IL-4 Tg mice vaccinated with nBCG produced the opposite profile. These findings suggest that BCG can be made more robust by incorporating immunopotentiating cytokines into the vaccine. PMID- 12096040 TI - Staphylococcal enterotoxin B-induced activation and concomitant resistance to cell death in CD28-deficient HLA-DQ8 transgenic mice. AB - HLA class II molecules present superantigens more efficiently than their murine counterpart. Therefore, transgenic mice expressing HLA-DQ8 with and without CD28 were used to address the role of CD28 in staphylococcal enterotoxin B (SEB) driven immune responses. SEB-induced in vitro proliferation of naive DQ8.CD28(-/ ) splenocytes was comparable to DQ8.CD28(+/+) cells, and was several fold higher than that of C57BL/10 and BALB/c splenocytes. SEB-activated, naive DQ8.CD28(-/-) cells in vitro produced significantly less IL-2, IL-4 and IL-10 than DQ8.CD28(+/+) cells, while IFN-gamma and IL-6 production was comparable. SEB induced in vivo expansion of CD4(+) T cells and, to a greater extent, CD8(+) T cells was compromised in DQ8.CD28(-/-) mice, indicating that SEB-induced proliferation of CD8(+) T cells is more dependent on CD28 co-stimulation. Upon re stimulation, SEB-primed CD28(+/+) T cells failed to proliferate but were capable of producing cytokines. Conversely, CD28(-/-) T cells were capable of proliferation, but not cytokine production. SEB-primed CD28-deficient cells produced significantly less nitric oxide when compared to CD28-sufficient cells following re-stimulation with SEB. CD28(+/+) and not CD28(-/-) mice were highly susceptible to SEB-induced lethal shock characterized by significantly elevated serum IFN-gamma. Thus, (i) efficient presentation of SEB by HLA-DQ8 circumvents co-stimulation through CD28, (ii) unique CD28-derived signals are mandatory for generation of certain effector functions, and (iii) absence of CD28-derived signals confers resistance to activation-induced cell death and protects mice from SEB-induced shock. PMID- 12096042 TI - Anti-beta(2)-glycoprotein I antibodies in children with atopic dermatitis. AB - beta(2)-Glycoprotein I (beta(2)GPI) appears to be the major antigen for antiphospholipid antibodies (aPL) in patients with antiphospholipid syndrome (APS). In early infancy, virtually all children initiate transient immune response to non-pathogenic nutritional antigens, which fails to terminate in children with atopic diseases. To examine the possibility that a prolonged immune response to beta(2)GPI could also spread to the human protein, antibodies against human beta(2)GPI (anti-beta(2)GPI) were determined in 93 randomly selected children with different allergic diseases. A high frequency (42%) of IgG anti beta(2)GPI was found in children with atopic dermatitis (AD), but not in those with other allergic diseases. Anti-beta(2)GPI in children with AD were exclusively of the IgG1 subclass and bound to bovine beta(2)GPI as well, but not to either beta(2)GPI combined with the phospholipid cardiolipin. The epitopes were identified in domain V of beta(2)GPI and the antibody binding was abolished upon the specific proteolytic cleavage of the phospholipid-binding C-terminal loop in domain V of beta(2)GPI. These results indicated that the epitopes for anti-beta(2)GPI in children with AD most likely resided in close vicinity of the phospholipid-binding site of beta(2)GPI. The epitopic difference from anti beta(2)GPI in APS may explain presumed non-thrombogenicity of anti-beta(2)GPI in children with AD. PMID- 12096041 TI - Molecular characterization of the mouse involuted thymus: aberrations in expression of transcription regulators in thymocyte and epithelial compartments. AB - Despite playing a critical role in the development of naive T cells, the thymus is involuted with age. Whether a single age-associated defect or multiple aberrations contribute to thymic involution remains controversial. Here, we determined molecular aberrations in the thymocyte and epithelium compartments of the aging thymus. We demonstrated that total thymocyte numbers declined with a stepwise kinetics; clear demarcations occurred at 1.5, 3, 12 and 22 months of age. By quantitative PCR, a 2.4-fold reduction in the copies of signal joint TCR excised circle (sjTREC)/10(5) thymocytes was first detected at 3 months; no further reduction observed thereafter. Nevertheless, the combined reductions in thymocyte numbers and sjTREC/10(5) cells caused a 7-fold decrease in sjTREC/thymus by 3 months, 21-fold by 18 months and 72-fold by 22 months as compared to 1 month. We showed aberration in expression of E2A, a transcription regulator critical for TCR beta rearrangement. While E2A expression declined 3 fold by 3 months and 18-fold by 7 months, expression of LMO2, a negative regulator of E2A activities, increased 5-fold by 18 months. Interestingly, expression of pre-T alpha and its transcriptional regulator HEB were not reduced with age. Furthermore, keratin-8 expression, specific for cortical thymic epithelium, declined 3-fold by 7 months and remained stable thereafter. In contrast, Foxn1 expression was reduced 3-fold by 3 months, 16-fold by 12 months and 37-fold by 18 months. IL-7 expression was not reduced until 7 months and reached 15-fold reduction by 22 months. Thus, the data demonstrate that thymic involution results not from a single defect, but culminates from an array of molecular aberrations in both the developing thymocytes and thymic epithelials. PMID- 12096043 TI - Water transport controversies--an overview. PMID- 12096045 TI - Transport of volatile solutes through AQP1. AB - For almost a century it was generally assumed that the lipid phases of all biological membranes are freely permeable to gases. However, recent observations challenge this dogma. The apical membranes of epithelial cells exposed to hostile environments, such as gastric glands, have no demonstrable permeability to the gases CO2 and NH3. Additionally, the water channel protein aquaporin 1 (AQP1), expressed at high levels in erythrocytes, can increase membrane CO2 permeability when expressed in Xenopus oocytes. Similarly, nodulin-26, which is closely related to AQP1, can act as a conduit for NH3. A key question is whether aquaporins, which are abundant in virtually every tissue that transports O2 and CO2 at high levels, ever play a physiologically significant role in the transport of small volatile molecules. Preliminary data are consistent with the hypothesis that AQP1 enhances the reabsorption of HCO3- by the renal proximal tubule by increasing the CO2 permeability of the apical membrane. Other preliminary data on Xenopus oocytes heterologously expressing the electrogenic Na+-HCO3- cotransporter (NBC), AQP1 and carbonic anhydrases are consistent with the hypothesis that the macroscopic cotransport of Na+ plus two HCO3- occurs as NBC transports Na+ plus CO3(2-) and AQP1 transports CO2 and H2O. Although data - obtained on AQP1 reconstituted into liposomes or on materials from AQP1 knockout mice - appear inconsistent with the model that AQP1 mediates substantial CO2 transport in certain preparations, the existence of unstirred layers or perfusion limited conditions may have masked the contribution of AQP1 to CO2 permeability. PMID- 12096046 TI - Does aquaporin-1 pass gas? An opposing view. PMID- 12096044 TI - Aquaporin water channels--from atomic structure to clinical medicine. AB - The water permeability of biological membranes has been a longstanding problem in physiology, but the proteins responsible for this remained unknown until discovery of the aquaporin 1 (AQP1) water channel protein. AQP1 is selectively permeated by water driven by osmotic gradients. The atomic structure of human AQP1 has recently been defined. Each subunit of the tetramer contains an individual aqueous pore that permits single-file passage of water molecules but interrupts the hydrogen bonding needed for passage of protons. At least 10 mammalian aquaporins have been identified, and these are selectively permeated by water (aquaporins) or water plus glycerol (aquaglyceroporins). The sites of expression coincide closely with the clinical phenotypes--ranging from congenital cataracts to nephrogenic diabetes insipidus. More than 200 members of the aquaporin family have been found in plants, microbials, invertebrates and vertebrates, and their importance to the physiology of these organisms is being uncovered. PMID- 12096047 TI - Analysis of the sodium recirculation theory of solute-coupled water transport in small intestine. AB - Our previous mathematical model of solute-coupled water transport through the intestinal epithelium is extended for dealing with electrolytes rather than electroneutral solutes. A 3Na+-2K+ pump in the lateral membranes provides the energy-requiring step for driving transjunctional and translateral flows of water across the epithelium with recirculation of the diffusible ions maintained by a 1Na+-1K+-2Cl- cotransporter in the plasma membrane facing the serosal compartment. With intracellular non-diffusible anions and compliant plasma membranes, the model describes the dependence on membrane permeabilities and pump constants of fluxes of water and electrolytes, volumes and ion concentrations of cell and lateral intercellular space (lis), and membrane potentials and conductances. Simulating physiological bioelectrical features together with cellular and paracellular fluxes of the sodium ion, computations predict that the concentration differences between lis and bathing solutions are small for all three ions. Nevertheless, the diffusion fluxes of the ions out of lis significantly exceed their mass transports. It is concluded that isotonic transport requires recirculation of all three ions. The computed sodium recirculation flux that is required for isotonic transport corresponds to that estimated in experiments on toad small intestine. This result is shown to be robust and independent of whether the apical entrance mechanism for the sodium ion is a channel, a SGLT1 transporter driving inward uphill water flux, or an electroneutral Na+-K+-2Cl- cotransporter. PMID- 12096048 TI - Solute recirculation. PMID- 12096050 TI - The presence of local osmotic gradients can account for the water flux driven by the Na+-glucose cotransporter. PMID- 12096049 TI - Water pumps. AB - The transport of water across epithelia has remained an enigma ever since it was discovered over 100 years ago that water was transported across the isolated small intestine in the absence of osmotic and hydrostatic pressure gradients. While it is accepted that water transport is linked to solute transport, the actual mechanisms are not well understood. Current dogma holds that active ion transport sets up local osmotic gradients in the spaces between epithelial cells, the lateral intercellular spaces, and this in turn drives water transport by local osmosis. In the case of the small intestine, which in humans absorbs about 8 l of water a day, there is no direct evidence for either local osmosis or aquaporin gene expression in enterocytes. Intestinal water absorption is greatly enhanced by glucose, and this is the basis for oral rehydration therapy in patients with secretory diarrhoea. In our studies of the intestinal brush border Na+-glucose cotransporter we have obtained evidence that there is a direct link between the transport of Na+, glucose and water transport, i.e. there is cotransport of water along with Na+ and sugar, that will account for about 50 % of the total water transport across the human intestinal brush border membrane. In this short review we summarize the evidence for water cotransport and propose how this occurs during the enzymatic turnover of the transporter. This is a general property of cotransporters and so we expect that this may have wider implications in the transport of water and other small polar molecules across cell membranes in animals and plants. PMID- 12096051 TI - Evidence against aquaporin-1-dependent CO2 permeability in lung and kidney. AB - AQP1-dependent CO2 transport has been suggested from the increased CO2 permeability in Xenopus oocytes expressing AQP1. Potential implications of this finding include AQP1-facilitated CO2 exchange in mammalian lung and HCO3-/CO2 transport in kidney proximal tubule. We reported previously that: (a) CO2 permeability in erythrocytes was not affected by AQP1 deletion, (b) CO2 permeability in liposomes was not affected by AQP1 reconstitution despite a 100 fold increased water permeability, and (c) CO2 blow-off by the lung in living mice was not impaired by AQP1 deletion. We extend these observations by direct measurement of CO2 permeabilities in lung and kidney. CO2 transport across the air-space-capillary barrier in isolated perfused lungs was measured from changes in air-space fluid pH in response to addition/removal of HCO3-/CO2 from the pulmonary artery perfusate. The pH was measured by pleural surface fluorescence of a pH indicator (BCECF-dextran) in the air-space fluid. Air-space fluid pH equilibrated rapidly (t(1/2) approximately 6 s) in response to addition/removal of HCO3-/CO2. However, the kinetics of pH change was not different in lungs of mice lacking AQP1, AQP5 or AQP1/AQP5 together, despite an up to 30-fold reduction in water permeability. CO2 transport across BCECF-loaded apical membrane vesicles from kidney proximal tubule was measured from the kinetics of intravesicular acidification in response to rapid mixing with a HCO3-/CO2 solution. Vesicles rapidly acidified (t(1/2) approximately 10 ms) in response to HCO3-/CO2 addition. However the acidification rate was not different in kidney vesicles from AQP1 null mice despite a 20-fold reduction in water permeability. The results provide direct evidence against physiologically significant transport of CO2 by AQP1 in mammalian lung and kidney. PMID- 12096052 TI - Mobility of ions, sugar, and water in the cytoplasm of Xenopus oocytes expressing Na(+)-coupled sugar transporters (SGLT1). AB - A model was set up to study water transport in membrane proteins expressed in Xenopus oocytes. The model was tested experimentally using human and rabbit Na+ glucose cotransporters (SGLT1), and was used to explain controversies regarding unstirred layer effects. Cotransport of Na+, sugar and water was monitored by two electrode voltage clamp and online measurements of oocyte volume. The specific resistance of the oocyte cytoplasm was found by means of microelectrodes to be 263 +/- 91 Omega cm (S.D., n = 52), or 2.5 times that of Kulori medium, in agreement with reported values of intracellular ion concentrations and diffusion constants. Osmotically induced volume and resistance changes were compatible with a model of the oocyte in which 37 +/- 17 % (S.D., n = 66) of the intracellular volume acts as a free solution while the remainder is inert, being occupied by organelles, etc. The model explains the results of several types of experiments: rapid changes in rates of water cotransport induced by changes in clamp voltage followed by osmotic equilibration in sugar-free conditions; volume changes induced by Na+ transport via the ionophore gramicidin; and uphill water transport. Ethanol (0.5 %) induced a marked swelling of the oocytes of about 16 pl x s(-1). If the specific inhibitor of SGLT1 phlorizin is added from stock solutions in ethanol, the effect of ethanol obfuscates the effects of the inhibitor. We conclude that the transport parameters derived for water cotransport by the SGLT1 can be attributed to the protein residing in the plasma membrane with no significant influences from unstirred layer effects. PMID- 12096053 TI - Functional characterisation and subcellular localisation of HCN1 channels in rabbit retinal rod photoreceptors. AB - Gating of voltage-dependent conductances in retinal photoreceptors is the first step of a process leading to the enhancement of the temporal performance of the visual system. The molecular components underlying voltage-dependent gating in rods are presently poorly defined. In the present work we have investigated the isoform composition and the functional characteristics of hyperpolarisation activated cyclic nucleotide-gated channels (HCN) in rabbit rods. Using immunocytochemistry we show the expression in the inner segment and cell body of the isoform 1 (HCN1). Electrophysiological investigations show that hyperpolarisation-activated currents (I(h)) can be measured only from the cell regions where HCN1 is expressed. Half-activation voltage (-75.0 +/- 0.3 mV) and kinetics (t(1/2) of 101 +/- 8 ms at -110 mV and 20 degrees C) of the I(h) in rods are similar to those of the macroscopic current carried by homomeric rabbit HCN1 channels expressed in HEK 293 cells. The homomeric nature of HCN1 channels in rods is compatible with the observation that cAMP induces a small shift (2.3 +/- 0.8 mV) in the half-activation voltage of I(h). In addition, the observation that within the physiological range of membrane potentials, cAMP does not significantly affect the gain of the current-to-voltage conversion, may reflect the need to protect the first step in the processing of visual signals from changes in cAMP turnover. PMID- 12096055 TI - Cholinergic modulation of the basal L-type calcium current in ferret right ventricular myocytes. AB - The effects of the cholinergic muscarinic agonist carbachol (CCh) on the basal L type calcium current, I(Ca,L), in ferret right ventricular (RV) myocytes were studied using whole cell patch clamp. CCh produced two major effects : (i) in all myocytes, extracellular application of CCh inhibited I(Ca,L) in a reversible concentration-dependent manner; and (ii) in many (but not all) myocytes, upon washout CCh produced a significant transient stimulation of I(Ca,L) ('rebound stimulation'). Inhibitory effects could be observed at 1 x 10(-10) M CCh. The mean steady-state inhibitory concentration-response relationship was shallow and could be described with a single Hill equation (maximum inhibition = 34.5 %, IC50 = 4 x 10(-8) M, Hill coefficient n = 0.60). Steady-state inhibition (1 or 10 microM CCh) had no significant effect on I(Ca,L) selectivity or macroscopic (i) activation characteristics, (ii) inactivation kinetics, (iii) steady-state inactivation or (iv) kinetics of recovery from inactivation. Maximal inhibition of nitric oxide synthase (NOS) activity (preincubation of myocytes in 1 mM L-NMMA (N(G)-monomethyl-L-arginine) + 1 mM L-NNA (N(G)-nitro-L-arginine) for 2-3 h plus inclusion of 1 mM L-NMMA + 1 mM L-NNA in the patch pipette solution) produced no significant attenuation of the CCh-mediated inhibition of I(Ca,L). Protocols involving (i) the nitric oxide (NO) scavenger PTIO (2-phenyl-4,4,5,5, tetramethylimidazoline-1-oxyl-3-oxide; 200 microM), (ii) imposition of a 'cGMP clamp' (100 microM 8-Bromo-cGMP), and (iii) inhibition of soluble guanylyl cyclase (ODQ (1H-[1,2,4,]oxadiazolo(4,3,-a)quinoxalin-1-one), 50 microM) all failed to attenuate CCh-mediated inhibition of I(ca,L). While CCh consistently inhibited basal I(Ca,L) in all RV myocytes studied, not all myocytes displayed rebound stimulation upon CCh washout. However, there was no difference between CCh-mediated inhibition of I(Ca,L) between these two RV myocyte types, and in myocytes displaying rebound stimulation neither ODQ nor 8-Bromo-cGMP (8-Br-cGMP) altered the effect. We conclude that NO production, activation of soluble guanylyl cyclase, or changes in intracellular cGMP levels are not obligatorily involved in muscarinic-mediated modulation of basal I(Ca,L) in ferret RV myocytes. PMID- 12096054 TI - AMPA and NMDA currents show different short-term depression in the dorsal lateral geniculate nucleus of the rat. AB - Paired-pulse depression was studied at the glutamatergic synapse between retinal afferents and thalamocortical cells in the rat dorsal lateral geniculate nucleus. The main objective of this study was to examine the contributions of the pre- and postsynaptic sites to this depression by comparing AMPA- and NMDA-receptor mediated responses. Equal depression of the two receptor components would indicate involvement of presynaptic mechanisms, while differences in depression would indicate involvement of postsynaptic mechanisms. Pharmacologically isolated AMPA- and NMDA-receptor-mediated currents were recorded using the whole-cell patch-clamp technique in acute thalamic slices. Both the AMPA and the NMDA components showed pronounced depression when retinal afferents were activated by paired pulses. The depression decayed within 5 s. The AMPA component was more strongly depressed than the NMDA component at paired-pulse intervals ranging from 20 to 200 ms, suggesting the involvement of postsynaptic mechanisms. For intervals of 500 ms and longer, the depression of the two components was identical, suggesting the involvement of purely presynaptic mechanisms. The degree of depression measured without the use of pharmacological tools produced similar results, thus excluding the involvement of presynaptic ionotropic glutamate receptors. Cyclothiazide, a blocker of AMPA-receptor desensitisation, reduced the difference in depression between the two components, suggesting that desensitisation of the AMPA receptors is a postsynaptic mechanism that contributes to the difference in depression between the AMPA and the NMDA components. PMID- 12096056 TI - KCNE4 is an inhibitory subunit to the KCNQ1 channel. AB - KCNE4 is a membrane protein belonging to a family of single transmembrane domain proteins known to have dramatic effect on the gating of certain potassium channels. However, no functional role of KCNE4 has been suggested so far. In the present paper we demonstrate that KCNE4 is an inhibitory subunit to KCNQ1 channels. Co-expression of KCNQ1 and KCNE4 in Xenopus oocytes completely inhibited the KCNQ1 current. This was reproduced in mammalian CHO-K1 cells. Experiments with delayed expression of mRNA coding for KCNE4 in KCNQ1-expressing oocytes suggested that KCNE4 exerts its effect on KCNQ1 channels already expressed in the plasma membrane. This notion was supported by immunocytochemical studies and Western blotting, showing no significant difference in plasma membrane expression of KCNQ1 channels in the presence or absence of KCNE4. The impact of KCNE4 on KCNQ1 was specific since no effect of KCNE4 could be detected if co-expressed with KCNQ2-5 channels or hERG1 channels. RT-PCR studies revealed high KCNE4 expression in embryos and adult uterus, where significant expression of KCNQ1 channels has also been demonstrated. PMID- 12096057 TI - Dual and opposing roles of presynaptic Ca2+ influx for spontaneous GABA release from rat medial preoptic nerve terminals. AB - Calcium influx into the presynaptic nerve terminal is well established as a trigger signal for transmitter release by exocytosis. By studying dissociated preoptic neurons with functional adhering nerve terminals, we here show that presynaptic Ca2+ influx plays dual and opposing roles in the control of spontaneous transmitter release. Thus, application of various Ca2+ channel blockers paradoxically increased the frequency of spontaneous (miniature) inhibitory GABA-mediated postsynaptic currents (mIPSCs). Similar effects on mIPSC frequency were recorded upon washout of Cd2+ or EGTA from the external solution. The results are explained by a model with parallel Ca2+ influx through channels coupled to the exocytotic machinery and through channels coupled to Ca2+ activated K+ channels at a distance from the release site. PMID- 12096058 TI - Functional characteristics of non-NMDA-type ionotropic glutamate receptor channels in AII amacrine cells in rat retina. AB - The properties of non-NMDA glutamate receptor channels in AII amacrine cells were studied by patch-clamp recording from rat retinal slices. Application of AMPA or kainate to intact cells evoked currents with no apparent desensitization (EC50 of 118 microM for AMPA and 169 microM for kainate). Application of AMPA to patches evoked desensitizing responses with an EC50 of 217 and 88 microM for the peak and steady-state responses, respectively. Kainate-evoked responses of patches displayed no desensitization (EC50 = 162 microM). Cyclothiazide strongly potentiated AMPA-evoked responses and the AMPA-receptor antagonist GYKI 53655 inhibited both AMPA- and kainate-evoked responses (IC50 = 0.5-1.7 microM). Pre equilibration with GYKI 53655 completely blocked the response to kainate and pretreatment with concanavalin A did not unmask a response mediated by kainate receptors. AMPA- and kainate-evoked currents reversed close to 0 mV. AMPA-evoked peak and steady-state response components in patches displayed linear and outwardly rectifying I-V relations with an RI (ratio of the slope conductances at +40 mV and -60 mV) of 0.96 +/- 0.11 and 5.6 +/- 1.3, respectively. AMPA-evoked currents displayed a voltage-dependent relaxation after steps to positive or negative membrane potentials, indicating that the outward rectification of the steady-state response is caused by a voltage-dependent kinetic parameter of channel gating. Under bi-ionic conditions ([Ca2+](out) = 30 mM, [Cs+)(in) = 171 mM), the reversal potentials of AMPA- and kainate-evoked currents indicated channels with significant Ca2+ permeability (P(Ca)/P(Cs) = 1.9-2.1). Stationary noise analysis indicated that kainate activated channels with an apparent chord conductance of approximately 9 pS. Non-stationary noise analysis indicated that AMPA and glutamate activated channels with apparent chord conductances of approximately 9, approximately 15, approximately 23 and approximately 38 pS. Discrete single-channel gating corresponding to chord conductances of approximately 23 pS could be directly observed in some responses. Thus, our results indicate expression of high-affinity, voltage-sensitive AMPA receptors with significant Ca2+ permeability and relatively large single-channel chord conductances in AII amacrine cells. PMID- 12096059 TI - Fast network oscillations induced by potassium transients in the rat hippocampus in vitro. AB - Brief pressure ejection of solutions containing potassium, caesium or rubidium ions into stratum radiatum of the CA1 or CA3 regions of the hippocampal slice evoked a fast network oscillation. The activity evoked lasted approximately 3-25 s with the predominant frequency component being in the gamma frequency range (30 80 Hz), although beta frequency (15-30 Hz) and ultrafast (> 80 Hz) components could also be seen. The gamma frequency component of the oscillation remained constant, even when large changes in power occurred, and was synchronous across the CA1 region. Measurements with potassium ion-sensitive electrodes revealed that the network oscillation was accompanied by increases (0.5 to 2.0 mM) in the extracellular potassium concentration [K+]o. Bath application of the N-methyl-D aspartate (NMDA) receptor antagonists D(-)-2-amino-5-phosphonopentanoic acid (D AP5; 50 microM) had no significant effect but the alpha-amino-3-hydroxy-5-methyl 4-isooxazolepropionic acid (AMPA)/kainate receptor antagonist 2,3,-dioxo-6-nitro 1,2,3,4-tetrahydrobenzo[f]quinoxaline-7-sulphonamide disodium (NBQX; 20 microM) caused a significant reduction (86.7 +/- 4.5 %) in the power in the gamma frequency range. Residual rhythmic activity, presumably arising in the interneuronal network, was blocked by the GABA(A) receptor antagonist bicuculline. The putative gap junction blocker octanol caused a decrease in the power of the gamma frequency component of 75.5 +/- 5.6 %, while carbenoxolone produced a reduction of only 14 +/- 42 %. These experiments demonstrate that a modest increase in exogenous [K+]o in the hippocampus in vitro is sufficient to evoke a fast network oscillation, which is an emergent property of the synaptically and electrically interconnected neuronal network. PMID- 12096060 TI - Effects of chloride transport on bistable behaviour of the membrane potential in mouse skeletal muscle. AB - The lumbrical skeletal muscle fibres of mice exhibited electrically bistable behaviour due to the nonlinear properties of the inwardly rectifying potassium conductance. When the membrane potential (V(m)) was measured continuously using intracellular microelectrodes, either a depolarization or a hyperpolarization was observed following reduction of the extracellular potassium concentration (K+o) from 5.7 mM to values in the range 0.76-3.8 mM, and V(m) showed hysteresis when K+o was slowly decreased and then increased within this range. Hypertonicity caused membrane depolarization by enhancing chloride import through the Na+-K+ 2Cl- cotransporter and altered the bistable behaviour of the muscle fibres. Addition of bumetanide, a potent inhibitor of the Na+-K+-2Cl- cotransporter, and of anthracene-9-carboxylic acid, a blocker of chloride channels, caused membrane hyperpolarization particularly under hypertonic conditions, and also altered the bistable behaviour of the cells. Hysteresis loops shifted with hypertonicity to higher K+o values and with bumetanide to lower values. The addition of 80 microM BaCl2 or temperature reduction from 35 to 27 degrees C induced a depolarization of cells that were originally hyperpolarized. In the K+o range of 5.7-22.8 mM, cells in isotonic media (289 mmol x kg(-1)) responded nearly Nernstianly to K+o reduction, i.e. 50 mV per decade; in hypertonic media this dependence was reduced to 36 mV per decade (319 mmol x kg(-1)) or to 31 mV per decade (340 mmol x kg( 1)). Our data can explain apparent discrepancies in DeltaV(m) found in the literature. We conclude that chloride import through the Na+-K+-2Cl- cotransporter and export through Cl- channels influenced the V(m) and the bistable behaviour of mammalian skeletal muscle cells. The possible implication of this bistable behaviour in hypokalaemic periodic paralysis is discussed. PMID- 12096062 TI - Potentiation of 'on' bipolar cell flash responses by dim background light and cGMP in dogfish retinal slices. AB - The high sensitivity of the vertebrate visual system results from amplification inherent in phototransduction in rods and from the amplification of rod signals on their synaptic transfer at the first synapse with 'on' bipolar cells. These cells possess a metabotropic glutamate receptor linked via a cGMP cascade to the control of cGMP-activated channels. In the study presented here, we show that very dim background light, isomerising only one rhodopsin in 1 out of 10 rods per second, potentiates 'on' bipolar cell responses to superimposed flashes. Responses to dim flashes, which were undetectable above the noise in the dark, were boosted above the increased noise level induced by the background. This potentiation could be reproduced by elevating cGMP, which increases with light, or by dialysing the cells with a non-hydrolysable cGMP analogue. Inhibition of tyrosine kinase activity also reproduced the effect and induced a speeding up of the rising phase of the flash response, similar to the action of dim background light. Conversely, inhibition of tyrosine phosphatase activity blocked the potentiation. These results suggest that cGMP promotes tyrosine-site dephosphorylation of 'on' bipolar cell cGMP-activated channels, resulting in a rise in the sensitivity to cGMP, as has recently been demonstrated for rod cGMP activated channels. This constitutes a positive feedback mechanism such that as cGMP increases with light, the sensitivity of the channels to cGMP increases and boosts the signal above background noise. This mechanism would allow stochastic resonance to occur, facilitating single-photon detection when dark-adapted, and may therefore lead to improved discrimination. PMID- 12096061 TI - Cell type dependence and variability in the short-term plasticity of EPSCs in identified mouse hippocampal interneurones. AB - Synapses exhibit different short-term plasticity patterns and this behaviour influences information processing in neuronal networks. We tested how the short term plasticity of excitatory postsynaptic currents (EPSCs) depends on the postsynaptic cell type, identified by axonal arborizations and molecular markers in the hippocampal CA1 area. Three distinct types of short-term synaptic behaviour (facilitating, depressing and combined facilitating-depressing) were defined by fitting a dynamic neurotransmission model to the data. Approximately 75 % of the oriens-lacunosum-moleculare (O-LM) interneurones received facilitating EPSCs, but in three of 12 O-LM cells EPSCs also showed significant depression. Over 90 % of the O-LM cells were immunopositive for somatostatin and mGluR1alpha and all tested cells were decorated by strongly mGluR7a positive axon terminals. Responses in eight of 12 basket cells were described well with a model involving only depression, but the other cells displayed combined facilitating depressing EPSCs. No apparent difference was found between the plasticity of EPSCs in cholecystokinin- or parvalbumin-containing basket cells. In oriens bistratified cells (O-Bi), two of nine cells showed facilitating EPSCs, another two depressing, and the remaining five cells combined facilitating-depressing EPSCs. Seven of 10 cells tested for somatostatin were immunopositive, but mGluR1alpha was detectable only in two of 11 tested cells. Furthermore, most O-Bi cells projected to the CA3 area and the subiculum, as well as outside the hippocampal formation. Postsynaptic responses to action potentials recorded in vivo from a CA1 place cell were modelled, and revealed great differences between and within cell types. Our results demonstrate that the short-term plasticity of EPSCs is cell type dependent, but with significant heterogeneity within all three interneurone populations. PMID- 12096064 TI - Excess recovery heat production by isolated muscles from mice overexpressing uncoupling protein-3. AB - Contractile and energetic performance of bundles of muscle fibres from the soleus of mice overexpressing uncoupling protein 3 (UCP-3tg) were compared with the performance of bundles from wild-type mice. Force and heat production were measured during a series of thirty 0.2 s isometric tetani at L(o), the length optimal for force. UCP-3tg fibres were as strong as the wild-type and maintained force in the series equally well; in the first tetanus force was 116.9 +/- 15.1 and 133.3 +/- 19.7 mN x mm(-2) respectively (all values means +/- S.E.M., n = 6 for UCP-3tg and n = 5 for wild-type). Heat production was partitioned into initial heat (due to contractile ATPases and the creatine kinase reaction) and recovery heat (due to other ATP-supplying processes) and expressed relative to the first cycle total heat. Initial heat production was similar for the UCP-3tg and wild-type fibres, decreasing during the series from 0.799 +/- 0.052 to 0.661 +/- 0.061 relative units (UCP-3tg), and from 0.806 +/- 0.024 to 0.729 +/- 0.039 relative units (wild-type). In both types the recovery heat was small at the start of the series and increased as the series progressed. At the end of the series, recovery heat production by UCP-3tg fibres, 1.575 +/- 0.246 relative units, was twice that of the wild-type fibres, 0.729 +/- 0.072 relative units. The extra recovery heat represents inefficient recovery in UCP-3tg fibres. This is the first direct evidence of enhanced energy dissipation as heat when UCP-3tg is overexpressed. PMID- 12096063 TI - Myosin light chain 2 modulates MgADP-induced contraction in rabbit skeletal and bovine cardiac skinned muscle. AB - Skinned skeletal and cardiac muscle fibres can be activated by MgADP in the presence of MgATP without Ca2+; the isometric tension is developed in a sigmoidal manner with the addition of MgADP under relaxing conditions. The critical concentrations of MgADP for this MgADP-induced contraction are about 7.5 and 2.6 mM for skeletal and cardiac muscle fibres, respectively. To investigate whether muscle regulatory proteins, myosin light chain 2 (LC2) and troponin C (TnC), play a part in the MgADP-induced contraction, these proteins were partly extracted by treatment with trans-1,2-cyclohexanediamine-N,N,N',N'-tetraacetic acid (CDTA), a chelater of divalent cations, and the MgADP-tension relationship was examined in rabbit psoas and bovine cardiac skinned fibres. We found that the sigmoidal MgADP tension relationship became hyperbolic after a partial extraction of LC2 (about 30 %) and TnC (about 70 %). Reconstitution with LC2 restored the sigmoidal MgADP tension relationship of control fibres almost fully in both skeletal and cardiac fibres, whereas reconstitution with TnC alone had no effect. Furthermore, cardiac fibres reconstituted with skeletal LC2 exhibited an MgADP-tension relationship intermediate between skeletal and cardiac fibres. The partial extraction of LC2 and TnC resulted in a reduction of the inhibitory effect of inorganic phosphate (P(i)) on the MgADP-activated tension. Reconstitution with LC2 restored the original P(i)-tension relationship, whereas reconstitution with TnC had no effect. In other words, extraction of LC2 apparently increased the affinity of myosin for MgADP but decreased the affinity for P(i). These results demonstrate that LC2 modulates MgADP-induced activation of actomyosin interaction. PMID- 12096065 TI - Secreted intestinal surfactant-like particles interact with cell membranes and extracellular matrix proteins in rats. AB - Surfactant-like particles (SLP) are secreted from enterocytes basolaterally into the lamina propria, and reach the apical surface through the intercellular tight junctions. Interactions of SLP with apical and basolateral membranes and with extracellular matrix proteins were measured using a solid-phase binding assay and gel overlays. Small-intestinal SLP bound to basolateral membranes much more than to apical membranes, and more tightly to fibronectin than to laminin (affinity constant K(a) = 1.23 x 10(-2) microg vs. 0.67 x 10(-2) microg; maximal number of binding sites 4.1 microg x ml(-1) vs. 0.32 microg x ml(-1)), but did not bind to collagen types I or IV. Small-intestinal SLP bound fibronectin more than colonic or gastric SLP. Binding to fibronectin was inhibited only partially by RGD peptide and gelatin, but not by heparin. An antibody against alpha(v) integrin also identified the fibronectin-binding component in SLP at approximately 220 kDa, which is the expected size for integrin heterodimers. SLP binding to apical microvillous membranes was weaker and was inhibited by heparin. SLP bound more strongly to heparin itself, and this binding was inhibited by glucuronic acid and chondroitin sulfate. These data are consistent with the hypothesis that the time spent by secreted SLP in the lamina propria is prolonged by strong interactions with proteins in the basolateral membranes, and in the intestinal lumen by weaker interactions with apical membrane components, including heparin. These interactions may allow SLP the time to exert their functions in each tissue compartment. PMID- 12096066 TI - Determination of alveolar epithelial cell phenotypes in fetal sheep: evidence for the involvement of basal lung expansion. AB - The factors that control the differentiation of alveolar epithelial cells (AECs) into type-I and type-II cells in vivo are largely unknown. As sustained increases in fetal lung expansion induce type-II AECs to differentiate into type-I cells, our aim was to determine whether reduced fetal lung expansion can induce type-I AECs to trans-differentiate into type-II AECs. Chronically catheterised fetal sheep were divided into two age-matched control groups and three experimental groups (n = 5 for each). The experimental groups were exposed to either: (1) 10 days of increased lung expansion induced by tracheal obstruction (TO), (2) 10 days of TO followed by 5 days of reduced lung expansion induced by lung liquid drainage (LLD), or (3) 10 days of TO followed by 10 days of LLD. Following 10 days of TO, 5 days of LLD reduced the proportion of type-I AECs from 89.4 +/- 0.9 % to 68.4 +/- 2.8 %, which was similar to control values (64.8 +/- 0.5 %), and increased the proportion of type-II AECs from 1.9 +/- 0.3 % to 21.9 +/- 2.8 %, which remained below control values (33.4 +/- 1.7 %). The same treatment increased surfactant protein (SP)-A, SP-B and SP-C mRNA levels (expressed as a percentage of control values) from 26.7 +/- 6.0 %, 40.0 +/- 7.3 % and 10.3 +/- 1.8 % to 78.1 +/- 10.3 %, 105.8 +/- 12.7 % and 121.0 +/- 14.1 %, respectively. Similar results were obtained after 10 days of LLD, which followed 10 days of TO. These results indicate that the phenotypes of type-I and type-II AECs are strongly influenced by the basal degree of lung expansion in fetal sheep. Furthermore, the coincident increase in type-II AEC proportions and SP mRNA levels in response to LLD suggests that type-I AECs can trans-differentiate into functional type-II cells, and hence are not terminally differentiated. PMID- 12096067 TI - Evidence for agonist-specific endothelial vasodilator dysfunction with ageing in healthy humans. AB - Endothelium-dependent vasodilatation declines with advancing age in humans independently of disease. The mechanisms responsible for this decline are not clear. We determined whether the age-related reduction in endothelium-dependent vasodilatation in response to acetylcholine reflects a specific agonist-related defect or rather a more general endothelial cell vasomotor abnormality. Twenty two young (23-35 years) and 41 older (50-76 years) healthy men were studied. Forearm blood flow (FBF) responses to intra-arterial infusions of acetylcholine, bradykinin, substance P, isoproterenol (isoprenaline) and sodium nitroprusside were measured by strain-gauge plethysmography. There were no differences in resting FBF between the young (3.9 +/- 0.2 ml (100 ml tissue)(-1) x min(-1)) and older men (4.0 +/- 0.2 ml (100 ml tissue)(-1) x min(-1)). The increase in FBF at the highest dose of acetylcholine was approximately 30 % lower (P < 0.01) in the older (from 4.0 +/- 0.2 to 12.3 +/- 0.7 ml (100 ml tissue)(-1) x min(-1)) compared with young men (from 3.9 +/- 0.2 to 17.1 +/- 1.5 ml (100 ml tissue)(-1) x min(-1)). In contrast to acetylcholine, the FBF responses to the other endothelial agonists were not impaired with age. The maximum increases in FBF in response to bradykinin (19.2 +/- 1.0 vs. 20.2 +/- 0.9 ml (100 ml tissue)(-1) x min(-1)), substance P (15.1 +/- 0.8 vs. 16.8 +/- 0.7 ml (100 ml tissue)(-1) x min(-1)) and isoproterenol (17.5 +/- 0.9 vs. 17.5 +/- 0.9 ml (100 ml tissue)(-1) x min(-1)) were not significantly different between the older and young subjects. There were no age-related differences in the FBF responses to sodium nitroprusside. These results demonstrate that, although acetylcholine-induced vasodilatation is impaired with age, forearm endothelial vasodilatation in response to bradykinin, substance P and isoproterenol are well preserved in healthy men. Moreover, these findings suggest that agonist-stimulated endothelium dependent vasodilatation is not universally impaired with age. PMID- 12096068 TI - Whole body and leg acetate kinetics at rest, during exercise and recovery in humans. AB - We have used a constant [1,2-(13)C]acetate infusion (0.12 micromol x min(-1) x kg( 1)) for 2 h at rest, followed by 2 h of one-legged knee-extensor exercise at 65% of leg maximal workload, and 3 h of recovery in six post-absorptive volunteers to quantify whole-body and leg acetate kinetics and determine whether the whole-body acetate correction factor can be used to correct leg substrate oxidation. The acetate whole-body rate of appearance (R(a)) was not significantly different at rest, during exercise or during recovery (365-415 micromol x min( 1)). The leg net acetate uptake was similar at rest and during recovery (approximately 10 micromol x min(-1)), but increased approximately 5-fold with exercise. At rest the leg acetate uptake (approximately 15 micromol x min(-1)) and release (approximately 5 micromol x min(-1)) accounted for 4 and 1.5 % of whole-body acetate disposal (R(d)) and R(a), respectively. When the leg acetate kinetics were extrapolated to the total body skeletal muscle mass, then skeletal muscle accounted for approximately 16 and approximately 6% of acetate R(d) and R(a). With exercise, leg acetate uptake increased approximately 6-fold, whereas leg acetate release increased 9-fold compared with rest. Whole-body acetate carbon recovery increased with time of infusion at rest and during recovery from 21% after 1.5 h of infusion to 45% in recovery after 7 h of infusion. Leg and whole-body acetate carbon recovery were similar under resting conditions, both before and after exercise. During exercise whole-body acetate carbon recovery was approximately 75%, however, acetate carbon recovery of the active leg was substantially higher (approximately 100%). It is concluded that inactive skeletal muscle plays a minor role in acetate turnover. However, active skeletal muscle enhances several-fold acetate uptake and subsequent oxidation, as well as release and its contribution to whole-body acetate turnover. Furthermore, under resting conditions the whole-body acetate correction factor can be used to correct for leg, skeletal muscle, substrate oxidation, but not during exercise. PMID- 12096071 TI - On organization of a neuronal network in pathways from group II muscle afferents in feline lumbar spinal segments. AB - The aim of the study was to verify the hypothesis that trisynaptic actions of group II muscle afferents upon motoneurones are, at least in part, mediated by dorsal horn interneurones exciting the same intermediate zone interneurones that are interposed in disynaptic pathways from group II afferents. Population EPSPs (field potentials) and responses of individual interneurones evoked by group II afferents in the dorsal horn and in the intermediate zone were analysed in order to assess the possibility of a causal relationship between them. When direct actions of group II afferents in the intermediate zone were abolished by presynaptic inhibition, distinct later components of field potentials and delayed interneuronal responses were induced at latencies 0.5-1 ms longer than those seen originally. Both the latency and a marked temporal facilitation define these later group II actions as being evoked disynaptically. Under the same conditions, single stimuli activated more than one half of dorsal horn interneurones, and the second and third stimuli activated all of these interneurones. Responses of dorsal horn interneurones preceded disynaptically evoked responses of intermediate zone interneurones. The study indicates that intermediate zone interneurones may be activated by group II afferents both directly and via dorsal horn interneurones and that synaptic actions of group II afferents upon these interneurones, and their subsequent actions upon motoneurones, may be modulated in parallel at the level of intermediate zone and dorsal horn interneurones. PMID- 12096070 TI - Differential presynaptic inhibition of actions of group II afferents in di- and polysynaptic pathways to feline motoneurones. AB - The aim of this study was to investigate differences in the effects of presynaptic inhibition of transmission from group II muscle afferents to neurones in the dorsal horn and in the intermediate zone and the consequences of these differences for reflex actions of group II afferents upon alpha-motoneurones. The degree of presynaptic inhibition was estimated from the degree of depression of monosynaptic components of population EPSPs (field potentials) evoked by group II muscle afferents in deeply anaesthetized cats. The decrease in the area of field potentials was considerably larger and longer lasting in the intermediate zone, where they were often obliterated, than in the dorsal horn, where they were reduced to about two-thirds. Presynaptic inhibition of field potentials evoked by other afferents at the same locations was much weaker. Intracellular records from alpha-motoneurones revealed that short latency EPSPs and IPSPs evoked from group II afferents are considerably reduced by conditioning stimuli that effectively depress intermediate zone field potentials of group II origin. The results of this study lead to the conclusion that strong presynaptic inhibition of transmission to intermediate zone interneurones allows a selective depression of disynaptic actions of group II muscle afferents on alpha- and gamma-motoneurones, mediated by these interneurones, and favours polysynaptic actions of these afferents. PMID- 12096069 TI - Melanin-concentrating hormone depresses L-, N-, and P/Q-type voltage-dependent calcium channels in rat lateral hypothalamic neurons. AB - Melanin-concentrating hormone (MCH), a cyclic 19-amino-acid peptide, is synthesized exclusively by neurons in the lateral hypothalamic (LH) area. It is involved in a number of brain functions and recently has raised interest because of its role in energy homeostasis. MCH axons and receptors are found throughout the brain. Previous reports set the foundation for understanding the cellular actions of MCH by using non-neuronal cells transfected with the MCH receptor gene; these cells exhibited an increase in cytoplasmic calcium in response to MCH, suggesting an excitatory action for the peptide. In the study presented here, we have used whole-cell recording in 117 neurons from LH cultures and brain slices to examine the actions of MCH. MCH decreased the amplitude of voltage dependent calcium currents in almost all tested neurons. The inhibition desensitized rapidly (18 s to half maximum at 100 nM concentration) and was dose dependent (IC(50) = 7.8 nM) when activated with a pulse from -80 mV to 0 mV. A priori activation of G-proteins with GTPgammaS completely eliminated the MCH induced effect at low MCH concentrations and reduced the MCH-induced effect at high MCH concentrations. Inhibition of G-proteins with pertussis toxin (PTX) blocked the MCH-induced inhibitory effect at high MCH concentrations. Pre-pulse depolarization resulted in an attenuation of the MCH-induced inhibition of calcium currents in most neurons. These data suggest that MCH exerts an inhibitory effect on calcium currents via PTX-sensitive G-protein pathways, probably the G(i)/G(o) pathway, in LH neurons. L-, N- and P/Q-type calcium channels were identified in LH neurons, with L- and N-type channels accounting for most of the voltage-activated current (about 40 % each); MCH attenuated each of the three types (mean 50 % depression), with the greatest inhibition found for N-type currents. In contrast to previous data on non-neuronal cells showing an MHC-evoked increase in calcium, our data suggest that the reverse occurs in LH neurons. The attenuation of calcium currents is consistent with an inhibitory action for the peptide in neurons. PMID- 12096072 TI - Renomedullary and intestinal hyaluronan content during body water excess: a study in rats and gerbils. AB - Our previous studies in rats have suggested a role for renomedullary hyaluronan (HA) in water homeostasis. The gerbil is known for its unique ability to conserve water. In the present study renal papillary and intestinal HA were compared between groups of anaesthetized gerbils and rats before and after up to 6 h of I.V. water loading. Baseline papillary HA in gerbils was only 37 % of that in the rat. Water loading in rats increased the papillary HA content. Elevation was maximal (+27 %, P < 0.05) after 2 h of water loading and then declined to control levels after 6 h of water loading (+3 %, n.s.). In contrast, the gerbil responded with a decreased papillary HA content during water loading. The depression was maximal after 2 h (-49 %, P < 0.05) and was still 41 % below the control values after 6 h (P < 0.05). The urine flow rate increased rapidly in the rat and its maximum, 21 times above the control level (P < 0.05), occurred at the HA peak, i.e. after 2 h of water loading while in the gerbil, the urine flow rate increased slowly and slightly and was only six times above control values after 6 h of water loading (P < 0.05). The HA content along the intestine was similar in the two species: lowest in the duodenum and jejunum and highest in the distal colon. To conclude, in the rat, the elevation of papillary interstitial HA during acute water loading would counteract water reabsorption by changing the physico chemical characteristics of the interstitial matrix favouring rapid water diuresis. This would work as a complement to the powerful regulation by ADH. The gerbil has a diametrically different regulation of papillary HA turnover during water loading. The decreased papillary HA level during water loading and the slow and small diuretic response may represent a genetic difference in adaptation to enhance the ability to conserve water in an arid environment. PMID- 12096073 TI - Bipedal distribution of human vestibular-evoked postural responses during asymmetrical standing. AB - Galvanic vestibular stimulation (GVS) evokes responses in muscles of both legs when bilateral stimuli are applied during normal stance. We have used this technique to assess whether asymmetrical standing alters the distribution of responses in the two legs. Subjects stood either asymmetrically with 75% of their body weight on one leg or symmetrically with each leg taking 50% of their body weight. The net response in each leg was taken from changes in ground reaction force measured from separate force plates under each foot. The net force profile consisted of a small initial force change that peaked at approximately 200 ms followed by an oppositely directed larger component that peaked at approximately 450 ms. We analysed the second force component since it was responsible for the kinematic response of lateral body sway and tilt towards the anode. In the horizontal plane, both legs produced lateral force responses that were in the same direction but larger in the leg ipsilateral to the cathodal ear. There were also vertical force responses that were of equal size in both legs but acted in opposite directions. When subjects stood asymmetrically the directions of the force responses remained the same but their magnitudes changed. The lateral force response became 2-3 times larger for the more loaded leg and the vertical forces increased 1.5 times on average for both legs. Control experiments showed that these changes could not be explained by either the consistent (< 5 deg) head tilt towards the side of the loaded leg or the changes in background muscle activity associated with the asymmetrical posture. We conclude that the redistribution of force responses in the two legs arises from a load-sensing mechanism. We suggest there is a central interaction between load-related afferent input from the periphery and descending motor signals from balance centres. PMID- 12096075 TI - Gene expression profiling of hereditary and sporadic ovarian cancers reveals unique BRCA1 and BRCA2 signatures. PMID- 12096077 TI - Will less drastic procedures replace radical prostatectomy? PMID- 12096074 TI - Understanding and appreciating overdiagnosis in the PSA era. PMID- 12096076 TI - High-intensity focused ultrasound makes its debut. PMID- 12096078 TI - Vagaries of research publishing again under the microscope. PMID- 12096079 TI - For the uninsured, health problems are more serious. PMID- 12096080 TI - Estimated excess deaths among uninsured adults, 2000. PMID- 12096081 TI - Retinoids and lung cancer: targeting the right population. PMID- 12096082 TI - Insulin, insulin-like growth factor-I (IGF-I), IGF binding proteins, their biologic interactions, and colorectal cancer. AB - Secular changes and worldwide variations in incidence rates of colorectal cancer, along with results from twin and migrant studies, provide compelling evidence that environmental factors influence the risk of this disease. Among the most important of these factors are diet and associated factors, such as physical activity and body size. Recent data suggest that dietary and related factors may influence colorectal cancer risk via their effects on serum insulin concentrations and on the bioavailability of insulin-like growth factor-I (IGF I). Epidemiologic studies have shown that IGF-I is positively associated with the risk of colorectal cancer, and experimental studies have shown that IGF-I has mitogenic and antiapoptotic actions on colorectal cancer cells. IGF-I bioactivity is regulated in part by its six binding proteins (IGFBP-1 to IGFBP-6); insulin inhibits the production of IGFBP-1 and perhaps IGFBP-2. As a result, chronically elevated fasting and postprandial insulin levels may lead to a decrease in circulating IGFBP-1 and IGFBP-2 concentrations and, consequently, an increase in IGF-I bioavailability. Insulin may also increase the circulating IGF-I/IGFBP-3 ratio by increasing hepatic growth hormone sensitivity. The increased IGF-I bioavailability may, over time, increase the risk of colorectal cancer. This new evidence for biologic interactions among insulin, IGF-I, and IGFBPs in the context of colorectal carcinogenesis provides a potential mechanism through which diet and associated factors may increase the risk of this cancer. PMID- 12096083 TI - Overdiagnosis due to prostate-specific antigen screening: lessons from U.S. prostate cancer incidence trends. AB - BACKGROUND: Overdiagnosis of clinically insignificant prostate cancer is considered a major potential drawback of prostate-specific antigen (PSA) screening. Quantitative estimates of the magnitude of this problem are, however, lacking. We estimated rates of prostate cancer overdiagnosis due to PSA testing that are consistent with the observed incidence of prostate cancer in the United States from 1988 through 1998. Overdiagnosis was defined as the detection of prostate cancer through PSA testing that otherwise would not have been diagnosed within the patient's lifetime. METHODS: We developed a computer simulation model of PSA testing and subsequent prostate cancer diagnosis and death from prostate cancer among a hypothetical cohort of two million men who were 60-84 years old in 1988. Given values for the expected lead time--that is, the time by which the test advanced diagnosis--and the expected incidence of prostate cancer in the absence of PSA testing, the model projected the increase in population incidence of prostate cancer associated with PSA testing. By comparing the model-projected incidence with the observed incidence derived from the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) registry data, we determined the lead times and corresponding overdiagnosis rates that were consistent with the observed data. RESULTS: SEER data on prostate cancer incidence from 1988 through 1998 were consistent with overdiagnosis rates of approximately 29% for whites and 44% for blacks among men with prostate cancers detected by PSA screening. CONCLUSIONS: Among men with prostate cancer that would be detected only at autopsy, these rates correspond to overdiagnosis rates of, at most, 15% in whites and 37% in blacks. The observed trends in prostate cancer incidence are consistent with considerable overdiagnosis among PSA-detected cases. However, the results suggest that the majority of screen-detected cancers diagnosed between 1988 and 1998 would have presented clinically and that only a minority of cases found at autopsy would have been detected by PSA testing. PMID- 12096084 TI - Gene expression profiles of BRCA1-linked, BRCA2-linked, and sporadic ovarian cancers. AB - BACKGROUND: Germline mutations in BRCA1 and BRCA2 are responsible for 5%-10% of epithelial ovarian cancers, but the molecular pathways affected by these mutations are unknown. We used complementary DNA (cDNA) microarrays to compare gene expression patterns in ovarian cancers associated with BRCA1 or BRCA2 mutations with gene expression patterns in sporadic epithelial ovarian cancers and to identify patterns common to both hereditary and sporadic tumors. METHODS: Tumor samples from 61 patients with pathologically confirmed epithelial ovarian adenocarcinoma with matched clinicopathologic features were studied, including 18 with BRCA1 founder mutations, 16 with BRCA2 founder mutations, and 27 without either founder mutation (termed sporadic cancers). The cDNA microarrays contained 7651 sequence-verified features. Gene expression data were analyzed with a modified two-sided F test, with P<.0001 considered statistically significant. The expression level of six genes was also studied with reverse transcription polymerase chain reaction. RESULTS: The greatest contrast in gene expression was observed between tumors with BRCA1 mutations and those with BRCA2 mutations; 110 genes showed statistically significantly different expression levels (P<.0001). This group of genes could segregate sporadic tumors into two subgroups, "BRCA1 like" and "BRCA2-like," suggesting that BRCA1-related and BRCA2-related pathways are also involved in sporadic ovarian cancers. Fifty-three genes were differentially expressed between tumors with BRCA1 mutations and sporadic tumors; six of the 53 mapped to Xp11.23 and were expressed at higher levels in tumors with BRCA1 mutations than in sporadic tumors. Compared with the immortalized ovarian surface epithelial cells used as reference, several interferon-inducible genes were overexpressed in the majority of tumors with a BRCA mutation and in sporadic tumors. CONCLUSIONS: Mutations in BRCA1 and BRCA2 may lead to carcinogenesis through distinct molecular pathways that also appear to be involved in sporadic cancers. Sporadic carcinogenic pathways may result from epigenetic aberrations of BRCA1 and BRCA2 or their downstream effectors. PMID- 12096085 TI - A randomized phase IIb trial of anethole dithiolethione in smokers with bronchial dysplasia. AB - BACKGROUND: Results from preclinical studies have suggested that the organosulfur compound anethole dithiolethione (ADT) may be an effective chemopreventive agent for lung cancer. We conducted a phase IIb study to determine the effects of ADT in smokers with bronchial dysplasia. METHODS: One hundred twelve current and former smokers with a smoking history of at least 30 pack-years and at least one site of bronchial dysplasia identified by an autofluorescence bronchoscopy directed biopsy were randomly assigned to receive placebo or ADT at 25 mg orally thrice daily for 6 months. Each subject then underwent a follow-up bronchoscopy directed biopsy. We used changes in histopathologic grade and nuclear morphometry index (MI) as the primary and secondary end point biomarkers, respectively. Chi square tests with continuity correction were used to compare response rates on a lesion- and person-specific basis between the two study groups. All statistical tests were two-sided. RESULTS: One hundred one subjects had a follow-up bronchoscopy. In the lesion-specific analysis, progression rate of pre-existing dysplastic lesions by two or more grades and/or the appearance of new lesions was statistically significantly lower in the ADT group (8%) than in the placebo group (17%) (P<.001; difference = 9%, 95% confidence interval [CI] = 4% to 15%). In the person-specific analysis, the disease progression rate was statistically significantly lower in the ADT group (32%) than in the placebo group (59%) (P =.013; difference = 27%, 95% CI = 6% to 48%). The two treatment groups did not differ statistically significantly in terms of nuclear MI. Among individuals with an abnormal nuclear MI before treatment (29 in the ADT group and 25 in the placebo group), the progression rate in the ADT group (41%) was substantially lower than that in the placebo group (60%), although the difference was not statistically significant (P =.28; difference = 19%, 95% CI = -11% to 49%). Adverse events were mostly minor gastrointestinal symptoms that resolved with dose reduction or discontinuation of the medication. CONCLUSION: Our results suggest that, in smokers, ADT is a potentially efficacious chemoprevention agent for lung cancer. PMID- 12096087 TI - Leukemic cell-surface CD13/aminopeptidase N and resistance to apoptosis mediated by endothelial cells. AB - BACKGROUND: Attachment of leukemic cells to vascular endothelial cells induces the vascular endothelial cells to release endothelial cell-derived interleukin 8 (endothelial IL-8), which then induces leukemic cells to undergo apoptosis. NB4, a human promyelocytic leukemic cell line that expresses high levels of cell surface CD13/aminopeptidase N, does not undergo endothelial IL-8-induced apoptosis. Consequently, we investigated the relationship between cell-surface aminopeptidase activity and endothelial IL-8 induction of apoptosis in various leukemic cell lines. METHODS: CD13/aminopeptidase N activity and IL-8-induced apoptosis were examined in leukemic cell lines. Endothelial IL-8-induced apoptosis was examined further in NB4 cells, K562 cells (human chronic myelogenous leukemic cells expressing low levels of CD13/aminopeptidase N), CD13/aminopeptidase N-transfected K562 (K562/CD13) cells that overexpress aminopeptidase, and mock-transfected K562 cells (vector only). These cells were also cocultured with a vascular endothelial cell layer to investigate the association between aminopeptidase activity and apoptosis in this system. All statistical tests were two-sided. RESULTS: Endothelial IL-8 induced apoptosis in K562 cells but not in K562/CD13 cells. A combination of an aminopeptidase inhibitor (such as bestatin) and endothelial IL-8 induced apoptosis in NB4 cells and K562/CD13 cells (2.88-fold difference [95% confidence interval [CI] = 1.82 fold to 3.94-fold], P =.004 for bestatin-treated NB4 cells and 4.31-fold difference [95% CI = 3.52-fold to 5.10-fold], P<.001 for bestatin-treated K562/CD13 cells). When aminopeptidase activity in NB4 cells was modulated by aminopeptidase inhibitors, a statistically significant correlation was found between aminopeptidase activity and the proportion of apoptotic cells induced by endothelial IL-8 (r = -.837, P<.001 by Pearson's correlation coefficient; r = .697, P =.013 by Spearman's correlation analysis by ranks). K562/CD13 cells cocultured with vascular endothelial cells did not undergo apoptosis, but the addition of bestatin resulted in the induction of apoptosis in K562/CD13 cells (2.70-fold difference [95% CI = 1.77-fold to 3.63-fold], P<.001). Bestatin treatment increased the level of IL-8 mRNA in and the amount of IL-8 secreted by vascular endothelial cells. CONCLUSIONS: High levels of cell-surface CD13/aminopeptidase N appear to allow leukemic cells to resist endothelial IL-8 induced apoptosis. The combination of endothelial IL-8 and bestatin induce leukemic cells expressing high levels of CD13/aminopeptidase N to undergo apoptosis. Bestatin may be useful for treating patients with leukemia. PMID- 12096086 TI - Effects of interferon beta on transcobalamin II-receptor expression and antitumor activity of nitrosylcobalamin. AB - BACKGROUND: The ubiquitous plasma membrane transcobalamin II receptor (TC II-R) mediates uptake of cobalamin (Cbl; vitamin B12), an essential micronutrient. Tumors often require more Cbl than normal tissue, and increased Cbl uptake may result from increased TC II-R expression. To examine whether Cbl could therefore be used as a carrier molecule to target a chemotherapy drug, we tested an analogue of Cbl with nitric oxide as a ligand, nitrosylcobalamin (NO-Cbl). Because interferon beta (IFN-beta) has antitumor effects and increases expression of some membrane receptors, we examined whether it may enhance the effects of NO Cbl. METHODS: Antiproliferative effects of NO-Cbl were assessed in 24 normal and cancer cell lines. Xenograft tumors of human ovarian cancer NIH-OVCAR-3 cells were established in athymic nude mice, and tumor growth was monitored after treatment with NO-Cbl and IFN-beta, both individually and concomitantly. TC II-R expression and apoptosis was monitored in vitro and in vivo. RNA protection assays and mitochondrial membrane potential assays were used to distinguish the extrinsic and intrinsic apoptotic pathways, respectively. RESULTS: Cancer cell lines were more sensitive to NO-Cbl (with ID(50)s [the dose that inhibits growth by 50%] as low as 2 microM) than normal cell lines (with ID(50)s of 85-135 microM). Single-agent NO-Cbl and IFN-beta treatment of NIH-OVCAR-3 xenografts induced tumor regression, whereas combination treatment induced tumor eradication. IFN-beta treatment increased TC II-R expression in vitro and uptake of [(57)Co]cobalamin in vivo. Compared with NIH-OVCAR-3 cells treated with NO Cbl, cells treated with NO-Cbl and IFN-beta were more apoptotic and expressed higher mRNA levels of various apoptosis-associated genes. No changes in mitochondrial membrane potential were observed in cells treated with NO-Cbl. CONCLUSION: NO-Cbl inhibited tumor growth in vivo by activating the extrinsic apoptotic pathway. The increased expression of TC II-R induced by IFN-beta resulted in enhanced antitumor effects with NO-Cbl both in vitro and in vivo. PMID- 12096088 TI - Re: Cisplatin-based therapy for elderly patients with advanced non-small-cell lung cancer: implications of Eastern Cooperative Oncology Group 5592, a randomized trial. PMID- 12096090 TI - Re: Blocking oncogenic Ras signaling for cancer therapy. PMID- 12096092 TI - Signs of the time: environmental input to the circadian clock. AB - The circadian clock forms one of the most fascinating adaptations to life on earth. Organisms can not only anticipate the day/night cycle but can make use of an internal clock to measure daylength as an indicator of the changing of the seasons. The innate period of the clock is not exactly equal to 24 h, but is reset each day by environmental signals at dawn and dusk, most notably by changes in light and temperature. This ability to re-entrain also ensures that the clock is synchronized with the day/night cycle which in turn is crucial for anticipation of dawn and dusk. Recent advances in the field have identified the photoreceptors involved in resetting the clock in several systems. This has revealed surprising similarities, but also key differences in the circadian systems of plants, fungi, insects, and mammals. One recurring feature emerging from this research is that the photoreceptors themselves are under the control of the clock with transcript abundance being tightly regulated. Furthermore, elements of a feedback pathway whereby the clock modulates the activity of the light input pathway are now being identified. PMID- 12096093 TI - MYB transcription factors in the Arabidopsis circadian clock. AB - The LATE ELONGATED HYPOCOTYL (LHY) and CIRCADIAN CLOCK ASSOCIATED (CCA1) genes encode closely related MYB transcription factors, which regulate circadian rhythms in Arabidopsis. LHY and CCA1 verify some of the properties of oscillator components, since (i) expression of their transcripts and protein exhibits circadian oscillations; (ii) their constitutive expression abolishes overt rhythmicity and (iii) they function as components of a negative transcriptional feedback loop. LHY and CCA1 have been proposed to function in conjunction with the pseudo response regulator TOC1, as components of the circadian oscillator. The regulation of their respective transcripts and protein levels in response to light signals suggests that these proteins may also mediate the regulation of circadian rhythms by light. This review discusses experimental evidence for these hypotheses. PMID- 12096094 TI - Esr proteins are secreted by the cells of the embryo surrounding region. AB - Three highly homologous Esr genes are expressed specifically in the embryo surrounding region at the micropylar end of the maize endosperm. The proteins belong to a family of small hydrophilic proteins that share a conserved motive with Clv3, the ligand of the receptor-like kinase Clv1. In this study, co localization of Esr proteins with their mRNAs in the embryo surrounding region was shown with polyclonal antibodies recognizing all three Esr proteins. On a subcellular level the secretion of Esr proteins and their association with the cell wall was shown independently by cell fractionation, immunohistochemistry and transient expression of Gfp fusion proteins. Furthermore, a possible interaction of Esr proteins with a 35 kDa protein present in the lower half of maize kernels was suggested by in vitro affinity chromatography. Therefore Esr proteins share two characteristics with ligands of receptor-like kinases: they are released in the extracellular space and they have the capacity to form protein-protein interactions. PMID- 12096095 TI - Effect of grain colour gene (R) on grain dormancy and sensitivity of the embryo to abscisic acid (ABA) in wheat. AB - The level of grain dormancy and sensitivity to ABA of the embryo, a key factor in grain dormancy, were examined in developing grains of a white-grained wheat line, Novosibirskaya 67 (NS-67), and its red-grained near-isogenic lines (ANK-1A to 1D); a red-grained line, AUS 1490, and its white-grained mutant line (EMS-AUS). ANK lines showed higher levels of grain dormancy than NS-67 at harvest maturity. AUS 1490 grain also showed higher dormancy than EMS-AUS grain. These results suggest that the R gene for grain colour can enhance grain dormancy. However, the dormancy effect conferred by the R gene was not large, suggesting that it plays a minor role in the development of grain dormancy. Water extracts of AUS 1490 and EMS-AUS bran contained germination inhibitors equivalent to 1-10 microM ABA, although there was no difference in the amount of inhibitors between AUS 1490 and EMS-AUS. Thus, the grain colour gene of AUS 1490 did not appear to enhance the level of grain dormancy by accumulating germination inhibitors in its bran. Sensitivity to ABA of embryos was higher in grains collected around harvest maturity for ANK lines and AUS 1490, compared with NS-67 and EMS-AUS. The R gene might enhance grain dormancy by increasing the sensitivity of embryos to ABA. PMID- 12096096 TI - Establishment of a reproducible tissue culture system for the induction of Arabidopsis somatic embryos. AB - Somatic embryogenesis is an example of totipotency and is used as a model system for studying embryogenesis. A reproducible tissue culture system was established for the large-scale induction of Arabidopsis somatic embryos. The method allows maintenance of high embryogenic competence over a one-year period. Using this tissue culture system, the expression of embryo-specific genes (ABI3, LEC1, FUS3) was detected in embryogenic cells and somatic embryos. Exogenous application of abscisic acid enhanced the expression of some late-embryogenesis-abundant (LEA) protein genes in somatic embryos. The experiments show that the method can be used to obtain sufficient amounts of embryogenic material for basic molecular analyses. PMID- 12096097 TI - Kinetic characteristics of chloroplast glucose transport. AB - Influx of labelled D-glucose into isolated spinach (Spinacia oleracea L. cv. Melody hybrid) chloroplasts was initially rapid followed by a period of slower influx. The stroma glucose concentration attained equilibrium rapidly with low external glucose concentrations and the two were linearly proportional. The period of slower influx resulted from conversion of glucose to acidic products that remained trapped in the chloroplast. As the external glucose concentration increased, the stroma glucose concentration increased less and less, attaining a maximal concentration of 72 mol m(-3). The maintenance of an equilibrium stroma glucose concentration lower than that in the external medium is evidence that plastid glucose efflux involves secondary active transport. The equilibrium stroma glucose concentration increased in response to light and protonophoric uncouplers. It is proposed that glucose efflux is coupled with a proton and the stroma glucose concentration equilibrates in response to the proton gradient across the membrane. To determine if glucose is a significant product of starch mobilization, chloroplasts were isolated from spinach leaves labelled with 14CO2 during the preceding light period. Chloroplasts degraded starch at the same rate as the intact leaf. Glucose, maltose, and isomaltose were the principal labelled products that appeared in the medium during starch mobilization. The glucose concentration in the chloroplast was 2 mol m(-3), which is similar to the measured Km for zero trans efflux. The data support the role of the glucose translocator as an important component in the pathway for sucrose synthesis at night. PMID- 12096098 TI - Functional diversity of arbuscular mycorrhizas extends to the expression of plant genes involved in P nutrition. AB - This study of functional diversity considers symbiotic associations between two plant species, Medicago truncatula and Lycopersicon esculentum, and seven species of arbuscular mycorrhizal fungi (AMF). The objective was to integrate physiological analyses with molecular techniques to test whether functional diversity between AMF species is not only apparent at the level of mycorrhiza formation, plant nutrient uptake and plant growth, but also at the molecular level as observed by variation in the root expression of plant genes involved in the plant's P-starvation response. The seven species of AMF varied widely in their influence on the root expression of MtPT2 and Mt4 from M. truncatula and LePT1 and TPSI1 from L. esculentum. At one extreme was Glomus mosseae, whereby its colonization of M. truncatula resulted in the greatest reduction in MtPT2 and Mt4 gene expression and the highest level of P uptake and growth, while at the other extreme was Gigaspora rosea, whereby colonization resulted in the highest levels of MtPT2 and Mt4 gene expression and the lowest P uptake and growth. The expression of LePT1 and TPSI1 within the roots of L. esculentum was low and relatively uniform across the seven mycorrhizas, reflecting the ability of this cultivar to maintain low and constant shoot P levels despite root colonization by a broad selection of AMF. This study extends current understanding of functional diversity and shows that plants can respond differently to AMF, not only at the level of colonization, nutrient uptake and growth, but also at the level of gene expression. PMID- 12096099 TI - Bud abortion in tulip bulbs studied by magnetic resonance imaging. AB - After storage and subsequent planting of flower bulbs, the flower bud frequently appears to be aborted. This physiological aberration is probably caused by a change in the water status of the bulb and may be initiated during storage. The development of bud abortion in tulip bulbs was studied during long-term dry storage of the bulbs at 5 degrees C. The anatomy of individual tulip bulbs was followed non-invasively with T2-weighted NMR imaging, which allowed the monitoring of the growth of the shoot and daughter bulbs. Quantitative maps of T1 and T2 relaxation times of individual bulbs were used to assess regional changes in the water status of different tissues. Parallel to the NMR measurements, bulbs were planted to assess the ultimate flower quality. Moreover, water content, osmolality of tissue sap and ion leakage of excised shoot and scale tissues were determined to obtain information about the water status and viability of the bulbs. Significant decreases during long-term storage were found in T1 and T2 relaxation times in the shoot and particularly in the stamens. An increase in the osmolality of tissue sap and the decrease in relaxation times in the shoot below a certain threshold value attained after 24 weeks of storage, could be indicative for the emergence of bud abortion in tulips. PMID- 12096100 TI - Ecophysiological analysis of genotypic variation in peach fruit growth. AB - Cultivated varieties generally differ greatly from wild genotypes of the same closely related species. However, the processes responsible for these differences have not been elucidated. To analyse variations in fruit mass, fruit growth was characterized in a peach cultivar, a wild related species non-cultivated, and four hybrids derived by crossing them. These genotypes offer a wide range of agronomic values. An ecophysiological model of peach fruit growth in dry mass was used. This model simulates carbon partitioning at the 'shoot-bearing fruit' level by considering three compartments: fruits, 1-year-old stems and leafy shoots. The experimental measurements showed considerable variation between genotypes for fruit mass at maturity, fruit growth and source activity. The parameters of the ecophysiological model for each genotype were estimated from experimental data,. The model made it possible to account for genotypic variations in fruit growth and for genotype x fruit load interactions. Using the model, it was shown that the main processes explaining fruit growth variations among the genotypes studied were differences in potential fruit growth. PMID- 12096101 TI - The gravitropic setpoint angle of dark-grown rye seedlings and the role of ethylene. AB - The orientation growth of coleoptiles of dark-grown seedlings of rye (Secale cereale L. cv. Marder II), when grown under various conditions, was analysed with respect to the gravivector ('gravitropic setpoint angle', GSA). Coleoptiles growing through moist vermiculite attain and maintain a GSA with an average of about 180 degrees, i.e. a vertical orientation. Seedlings growing uncovered either on the surface of vermiculite or positionally fixed on filter paper attain and maintain a GSA of 140-150 degrees (i.e. deviating from the vertical by an average of 30-40 degrees ). Changing the position of the embryo relative to the horizontally fixed seed kernel or of the angle of the seed with respect to gravity during germination (+/-40 degrees relative to the horizontal) had no significant effect on the subsequent GSA of both covered and uncovered seedlings. The GSA of uncovered coleoptiles could be restored close to 180 degrees by treatment of the seedlings with ethylene, either applied via ethephon or 1 aminocyclopropane-1-carboxylic acid (ACC) as well as by fruit-released ethylene. The results are discussed with respect to the mechanism of the regulation of gravitropic growth of grass seedlings. PMID- 12096102 TI - The contribution of roots and shoots to whole plant nitrate reduction in fast- and slow-growing grass species. AB - The hypothesis was tested that slow-growing grass species perform a greater proportion of total plant NO3- reduction in their roots than do fast-growing grasses. Eight grass species were selected that differed in maximum relative growth rate (RGR) and net NO3- uptake rate (NNUR). Plants were grown with free access to nutrients in hydroponics under controlled-environment conditions. The site of in vivo NO3- reduction was assessed by combining in vivo NO3- reductase activity (NRA) assays with biomass allocation data, and by analysing the NO3- to amino acid ratio of xylem sap. In vivo NRA of roots and shoots increased significantly with increasing NNUR and RGR. The proportion of total plant NO3- reduction that occurs in roots was found to be independent of RGR and NNUR, with the shoot being the predominant site of NO3- reduction in all species. The theoretical maximum proportion of whole plant nitrogen assimilation that could take place in the roots was calculated using information on root respiration rates, RGR, NNUR, and specific respiratory costs associated with growth, maintenance and ion uptake. The calculated maximum proportion that the roots can contribute to total plant NO3- reduction was 0.37 and 0.23 for the fast-growing Dactylis glomerata L. and the slow-growing Festuca ovina L., respectively. These results indicate that slow-growing grass species perform a similar proportion of total plant NO3- reduction in their roots to that exhibited by fast-growing grasses. Shoots appear to be the predominant site of whole plant NO3- reduction in both fast- and slow-growing grasses when plants are grown with free access to nutrients. PMID- 12096103 TI - Peroxidase activity develops in the micropylar endosperm of tomato seeds prior to radicle protrusion. AB - Peroxidase activity developed specifically in the micropylar region of the endosperm of imbibed tomato seeds prior to radicle emergence. The activity was first detected approximately 24 h after the start of imibibition (6 h before radicle emergence) and increased markedly thereafter. In the lateral portion of the endosperm, peroxidase activity was undetectable for the first 2 d after the start of imbibition. Although the activity in the lateral endosperm became detectable 3 d after imbibition, the extent of the development of the activity was slight. The localization of peroxidase activity in the micropylar endosperm 2 d after the start of imbibition was confirmed by tissue printing analyses. When the endosperm tissues were wounded, there was an enhancement of the enzyme activity at the wounded region. H2O2 was formed at the expense of NADH only in the presence of Mn2+ and dinitrophenol by the extract from the micropylar endosperm in which peroxidase activity was present. The presence of H2O2 in the micropylar portion of the endosperm was shown histochemically. The possible functions of the peroxidases that develop in the endosperm of tomato seeds are discussed. PMID- 12096104 TI - Light-regulated leaf expansion in two Populus species: dependence on developmentally controlled ion transport. AB - Leaf growth responses to light have been compared in two species of Populus, P. deltoides and P. trichocarpa. These species differ markedly in morphology, anatomy, and dependence on light during leaf expansion. Light stimulates the growth rate and acidification of cell walls in P. trichocarpa but not in P. deltoides, whereas leaves of P. deltoides maintain growth in the dark. Light induced growth is promoted in P. deltoides when cells are provided 50-100 mM KCl. In both species, light initially depolarizes, then hyperpolarizes mesophyll plasma membranes. However, in the dark, the resting E(m) of mesophyll cells in P. deltoides, but not in P. trichocarpa, is relatively insensitive to decade changes in external [K+]. Results suggest that light-stimulated leaf growth depends on developmentally regulated cellular mechanisms controlling ion fluxes across the plasma membrane. These developmental differences underlie species-level differences in growth and physiological responses to the photoenvironment. PMID- 12096105 TI - The fungal sheath of ectomycorrhizal pine roots: an apoplastic barrier for the entry of calcium, magnesium, and potassium into the root cortex? AB - The apoplastic permeability of the fungal sheath of two different ectomycorrhizal associations of Pinus sylvestris L. was analysed by laser microprobe mass analysis (LAMMA) and energy-dispersive X-ray spectroscopy (EDXS) after stable isotope labelling with 25Mg, 41K and 44Ca. Entry of 25Mg and 44Ca into the outer cortical apoplast of non-mycorrhizal roots was detected after 4 min of labelling. After a longer exposure time the endodermis with its Casparian band acted as an efficient apoplastic diffusion barrier for the radial movement of 25Mg and 44Ca into the stele. A fraction of approximately one-third of the apoplastic cations of the root cortex could not be exchanged against the external label even after longer exposure times. The ectomycorrhizal sheath of the two fungal species used, Pisolithus tinctorius (Pers.) Coker & Couch and Suillus bovinus (L. ex Fr.) Kuntze, does not completely inhibit the apoplastic movement of ions into the mycorrhizal root cortex, but retarded the penetration of isotopes into the cortical apoplast. In roots inoculated with S. bovinus, a clear labelling of the cortical apoplast could first be detected after 24 h of exposure to the stable isotope solution. At this time the labelling of the cortical apoplast in these mycorrhizal roots was higher than those of non-mycorrhizal roots and, with EDXS, changes in the element composition of the apoplast were detected. The results indicated that possibly hydrophobins localized in the fungal cell wall might be involved in the increased hydrophobicity of mycorrhizal roots and the lower permeability of the ectomycorrhizal sheath. PMID- 12096106 TI - Source-sink coupling in young barley plants and control of phloem loading. AB - Phloem loading of carbohydrate within a mature exporting leaf of a barley seedling is shown to respond quickly to a change in the temperature of the root and the shoot meristem. This is interpreted as a close coupling between source supply and sink demand for carbohydrate, through the hydrostatic pressure gradient linking source and sink generated by the solute concentration within the sieve tubes. This interpretation was tested by using anoxia to alter solute concentration within the sieve tubes of one region of a leaf while observing phloem loading in an adjacent region. Responses to anoxia could not be explained by the above model, suggesting that either this model is incorrect or other signalling pathways are involved. There is evidence in the literature for coarse control of phloem loading but no evidence was found of fine control by solute content of the loaded sieve elements. PMID- 12096108 TI - Arbuscular mycorrhizal symbiosis regulates plasma membrane H+-ATPase gene expression in tomato plants. AB - Regulation by arbuscular mycorrhizal symbiosis of three tomato plasma membrane H+ ATPase genes (LHA1, LHA2 and LHA4) has been analysed in wild-type and mycorrhiza defective tomato plants. Expression of these genes was differentially regulated in leaves and roots of both tomato phenotypes after inoculation with Glomus mosseae. PMID- 12096107 TI - Cloning an iron-regulated metal transporter from rice. AB - Rice cDNA and genomic libraries were screened in order to clone an Fe(II) transporter gene. A cDNA clone highly homologous to the Arabidopsis Fe(II) transporter gene IRT1 was isolated from Fe-deficient rice roots. The cDNA clone was named OsIRT1. A genomic clone corresponding to the cDNA was also obtained, sequenced and analysed. When expressed in yeast cells, OsIRT1 cDNA reversed the growth defects of the yeast iron-uptake mutant. Northern blot analysis revealed that OsIRT1 mRNA was predominantly expressed in roots and was induced by Fe- and Cu-deficiency. This suggests that OsIRT1 is a functional metal transporter for iron, and is actively engaged in Fe uptake from soils, especially under limiting conditions. PMID- 12096109 TI - Proteomic strategies to reveal tumor heterogeneity among urothelial papillomas. AB - Proteomics and immunohistochemistry were used to reveal tumor heterogeneity among urothelial papillomas (UPs) with the long term goal of predicting their biological potential in terms of outcome. First, we identified proteins that were deregulated in invasive fresh lesions as compared with normal urothelium, and thereafter we immunostained UPs with a panel of antibodies against some of the markers. Twenty-two major proteins showing variations of 2-fold or more in at least one-third of the invasive lesions were selected. Specific antibodies against several of the proteins were obtained, but only a few reacted positively in immunostaining. A panel consisting of antibodies against keratinocytes (CKs) 5, 13, 18, and 20 and markers of squamous metaplasia (CKs 7, 8, and 14) was used to probe normal urothelium and 30 UPs collected during a period of five years. Four UPs showed a normal phenotype, whereas the rest could be grouped in five major types that shared aberrant staining with the CK20 antibody. Type 1 heterogeneity (n = 4) showed preferred staining of the umbrella cells with the CK8 antibody. Type 2 (n = 11) was typified by the staining of the basal and intermediate layers with the CK20 antibody. Type 3 (n = 7) was characterized by the predominant staining of the basal cell layer with the CK5 antibody. Type 4 (n = 1) showed areas of CK7 negative cells, whereas type 5 (n = 3) showed loss of staining of the basal cells with the CK20. 29% of the patients experienced recurrences, but none progressed to invasive disease. Patients harboring phenotypic alterations in the basal cell compartment (types 3 and 5) showed the highest number of recurrences (4/7 and 2/3, respectively), and all type 3 lesions progressed to a higher degree of dedifferentiation. Even though a long term prospective study involving a larger sample size is required to assess the biological potential of these lesions, we believe that this approach will prove instrumental for revealing early phenotypic changes in different types of cancer. PMID- 12096110 TI - Differential proteome analysis of replicative senescence in rat embryo fibroblasts. AB - Normal somatic cells undergo a finite number of divisions and then cease dividing whereas cancer cells are able to proliferate indefinitely. To identify the underlying mechanisms that limit the mitotic potential, a two-dimensional differential proteome analysis of replicative senescence in serially passaged rat embryo fibroblasts was undertaken. Triplicate independent two-dimensional gels containing over 1200 spots each were run, curated, and analyzed. This revealed 49 spots whose expression was altered more than 2-fold. Of these, 42 spots yielded positive protein identification by mass spectrometry comprising a variety of cytoskeletal, heat shock, and metabolic proteins, as well as proteins involved in trafficking, differentiation, and protein synthesis, turnover, and modification. These included gelsolin, a candidate tumor suppressor for breast cancer, and alpha-glucosidase II, a member of the family of glucosidases that includes klotho; a defect in klotho expression in mice results in a syndrome that resembles human aging. Changes in expression of TUC-1, -2, -4, and -4 beta, members of the TUC family critical for neuronal differentiation, were also identified. Some of the identified changes were also shown to occur in two other models of senescence, premature senescence of REF52 cells and replicative senescence of mouse embryo fibroblasts. The majority of these candidate proteins were unrecognized previously in replicative senescence. They are now implicated in a new role. PMID- 12096111 TI - Evidence that light modulates protein nitration in rat retina. AB - As part of ongoing efforts to better understand the role of protein oxidative modifications in retinal pathology, protein nitration in retina has been compared between rats exposed to damaging light or maintained in the dark. In the course of the research, Western methodology for detecting nitrotyrosine-containing proteins has been improved by incorporating chemical reduction of nitrotyrosine to aminotyrosine, allowing specific and nonspecific nitrotyrosine immunoreactivity to be distinguished. A liquid chromatography MS/MS detection strategy was used that selects all possible nitrotyrosine peptides for MS/MS based on knowing the protein identity. Quantitative liquid chromatography MS/MS analyses with tetranitromethane-modified albumin demonstrated the approach capable of identifying sites of tyrosine nitration with detection limits of 4-33 fmol. Using two-dimensional gel electrophoresis, Western detection, and mass spectrometric analyses, several different nitrotyrosine-immunoreactive proteins were identified in light-exposed rat retina compared with those maintained in the dark. Immunocytochemical analyses of retina revealed that rats reared in darkness exhibited more nitrotyrosine immunoreactivity in the photoreceptor outer segments. After intense light exposure, immunoreactivity decreased in the outer segments and increased in the photoreceptor inner segments and retinal pigment epithelium. These results suggest that light modulates retinal protein nitration in vivo and that nitration may participate in the biochemical sequela leading to light-induced photoreceptor cell death. Furthermore, the identification of nitrotyrosine-containing proteins from rats maintained in the dark, under non pathological conditions, provides the first evidence of a possible role for protein nitration in normal retinal physiology. PMID- 12096112 TI - Discordant protein and mRNA expression in lung adenocarcinomas. AB - The relationship between gene expression measured at the mRNA level and the corresponding protein level is not well characterized in human cancer. In this study, we compared mRNA and protein expression for a cohort of genes in the same lung adenocarcinomas. The abundance of 165 protein spots representing 98 individual genes was analyzed in 76 lung adenocarcinomas and nine non-neoplastic lung tissues using two-dimensional polyacrylamide gel electrophoresis. Specific polypeptides were identified using matrix-assisted laser desorption/ionization mass spectrometry. For the same 85 samples, mRNA levels were determined using oligonucleotide microarrays, allowing a comparative analysis of mRNA and protein expression among the 165 protein spots. Twenty-eight of the 165 protein spots (17%) or 21 of 98 genes (21.4%) had a statistically significant correlation between protein and mRNA expression (r > 0.2445; p < 0.05); however, among all 165 proteins the correlation coefficient values (r) ranged from -0.467 to 0.442. Correlation coefficient values were not related to protein abundance. Further, no significant correlation between mRNA and protein expression was found (r = 0.025) if the average levels of mRNA or protein among all samples were applied across the 165 protein spots (98 genes). The mRNA/protein correlation coefficient also varied among proteins with multiple isoforms, indicating potentially separate isoform-specific mechanisms for the regulation of protein abundance. Among the 21 genes with a significant correlation between mRNA and protein, five genes differed significantly between stage I and stage III lung adenocarcinomas. Using a quantitative analysis of mRNA and protein expression within the same lung adenocarcinomas, we showed that only a subset of the proteins exhibited a significant correlation with mRNA abundance. PMID- 12096113 TI - A strategy for the rapid identification of phosphorylation sites in the phosphoproteome. AB - Edman phosphate ((32)P) release sequencing provides a high sensitivity means of identifying phosphorylation sites in proteins that complements mass spectrometry techniques. We have developed a bioinformatic assessment tool, the cleavage of radiolabeled protein (CRP) program, which enables experimental identification of phosphorylation sites via (32)P labeling and Edman degradation of cleaved proteins obtained at femtomole levels. By observing the Edman cycle(s) in which radioactivity is found, candidate phosphorylation sites are identified by determining which residues occur at the observed number of cycles downstream from a peptide cleavage site. In cases where more than one residue could be responsible for the observed radioactivity, additional experiments with cleavage reagents having alternative specificities may resolve the ambiguity. Given a protein sequence and a cleavage site, CRP performs these experiments in silico, identifying resolved sites based on user-supplied experimental data, as well as suggesting combinations of reagents for additional analyses. Analysis of the PhosphoBase protein sequence database suggests that CRP data from two cleavage experiments can be used to identify unambiguously 60% of known phosphorylation sites. Data from additional cleavage experiments may increase the overall coverage to 70% of known sites. By comparing theoretical data obtained from the CRP program with (32)P release data obtained from an Edman sequencer, a known phosphorylation site was identified unambiguously and correctly. In addition, our results show that in vivo phosphorylation sites can be determined routinely by differential proteolysis analysis and Edman cycling with less than 1 fmol of protein and 1000 cpm. PMID- 12096114 TI - Complementary profiling of gene expression at the transcriptome and proteome levels in Saccharomyces cerevisiae. AB - Using an integrated genomic and proteomic approach, we have investigated the effects of carbon source perturbation on steady-state gene expression in the yeast Saccharomyces cerevisiae growing on either galactose or ethanol. For many genes, significant differences between the abundance ratio of the messenger RNA transcript and the corresponding protein product were observed. Insights into the perturbative effects on genes involved in respiration, energy generation, and protein synthesis were obtained that would not have been apparent from measurements made at either the messenger RNA or protein level alone, illustrating the power of integrating different types of data obtained from the same sample for the comprehensive characterization of biological systems and processes. PMID- 12096115 TI - Protein structure comparison using bipartite graph matching and its application to protein structure classification. AB - A measure of protein structure similarity is calculated from the matching of pairs of secondary structure elements between two proteins. The interaction of each pair was estimated from their axial line segments and combined with other geometric features to produce an optimal discrimination between intrafamily and interfamily relationships. The matching used a fast bipartite graph-matching algorithm that avoids the computational complexity of searching for the full subgraph isomorphism between the two sets of interactions. The main algorithm used was the "stable marriage" algorithm, which works on the ranked "preferences" of one interaction for another. The method takes 1/10 of a second for a typical comparison making it suitable as a fast pre-filter for slower, more exhaustive approaches. An application to protein structure classification is described. PMID- 12096116 TI - Proteomics - boom or bust? PMID- 12096117 TI - The Rickettsia prowazekii invasion gene homolog (invA) encodes a Nudix hydrolase active on adenosine (5')-pentaphospho-(5')-adenosine. AB - The genomic sequence of Rickettsia prowazekii, the obligate intracellular bacterium responsible for epidemic typhus, reveals an uncharacterized invasion gene homolog (invA). The deduced protein of 18,752 Da contains a Nudix signature, the specific motif found in the Nudix hydrolase family. To characterize the function of InvA, the gene was cloned and overexpressed in Escherichia coli. The expressed protein was purified to near homogeneity and subsequently tested for its enzymatic activity against a series of nucleoside diphosphate derivatives. The purified InvA exhibits hydrolytic activity toward dinucleoside oligophosphates (Np(n)N; n > or = 5), a group of cellular signaling molecules. At optimal pH 8.5, the enzyme actively degrades adenosine (5')-pentaphospho-(5') adenosine into ATP and ADP with a K(m) of 0.1 mM and k(cat) of 1.9 s(-1). Guanosine (5')-pentaphospho-(5')-guanosine and adenosine-(5')-hexaphospho (5') adenosine are also substrates. Similar to other Nudix hydrolases, InvA requires a divalent metal cation, Mg(2+) or Zn(2+), for optimal activity. These data suggest that the rickettsial invasion protein likely plays a role in controlling the concentration of stress-induced dinucleoside oligophosphates following bacterial invasion. PMID- 12096118 TI - Mass spectrometry-based methods for phosphorylation site mapping of hyperphosphorylated proteins applied to Net1, a regulator of exit from mitosis in yeast. AB - Prior to anaphase in Saccharomyces cerevisiae, Cdc14 protein phosphatase is sequestered within the nucleolus and inhibited by Net1, a component of the RENT complex in budding yeast. During anaphase the RENT complex disassembles, allowing Cdc14 to migrate to the nucleus and cytoplasm where it catalyzes exit from mitosis. The mechanism of Cdc14 release appears to involve the polo-like kinase Cdc5, which is capable of promoting the dissociation of a recombinant Net1.Cdc14 complex in vitro by phosphorylation of Net1. We report here the phosphorylation site mapping of recombinant Net1 (Net1N) and a mutant Net1N allele (Net1N-19m) with 19 serines or threonines mutated to alanine. A variety of chromatographic and mass spectrometric-based strategies were used, including immobilized metal affinity chromatography, alkaline phosphatase treatment, matrix-assisted laser desorption post-source decay, and a multidimensional electrospray mass spectrometry-based approach. No one approach was able to identify all phosphopeptides in the tryptic digests of these proteins. Most notably, the presence of a basic residue near the phosphorylated residue significantly hampered the ability of alkaline phosphatase to hydrolyze the phosphate moiety. A major goal of research in proteomics is to identify all proteins and their interactions and post-translational modification states. The failure of any single method to identify all sites in highly phosphorylated Net1N, however, raises significant concerns about how feasible it is to map phosphorylation sites throughout the proteome using existing technologies. PMID- 12096119 TI - A distinct repertoire of autoantibodies in hepatocellular carcinoma identified by proteomic analysis. AB - Chronic infections with hepatitis B (HBV) and hepatitis C (HCV) viruses are major risk factors for hepatocellular carcinoma (HCC). We have utilized a proteomic approach to determine whether a distinct repertoire of autoantibodies can be identified in HCC. Sera from 37 patients with HCC and 31 subjects chronically infected with HBV or HCV without HCC were investigated. Sera from 116 patients with other cancers, three patients with systemic lupus erythematosus, and 24 healthy subjects were utilized as controls. We report the identification of eight proteins, for each of which autoantibodies were detected in sera from more than 10% of patients with HCC but not in sera from healthy individuals (p < 0.05). Autoantibodies to four of these proteins were detected at a comparable frequency in sera from patients with chronic hepatitis. The other four proteins, which consisted of calreticulin isoforms, cytokeratin 8, nucleoside diphosphate kinase A, and F(1)-ATP synthase beta-subunit, induced autoantibodies among patients with HCC, independently of their HBV/HCV status. Calreticulin, and a novel truncated form of calreticulin (Crt32) we have identified, most commonly elicited autoantibodies among patients with HCC (27%). We conclude that a distinct repertoire of autoantibodies is associated with HCC that may have utility in early diagnosis of HCC among high risk subjects with chronic hepatitis. PMID- 12096120 TI - Deciphering protein complexes and protein interaction networks by tandem affinity purification and mass spectrometry: analytical perspective. AB - We employed a combination of tandem affinity purification and mass spectrometry for deciphering protein complexes and the protein interaction network in budding yeast. 53 genes were epitope-tagged, and their interaction partners were isolated by two-step immunoaffinity chromatography from whole cell lysates. 38 baits pulled down a total of 220 interaction partners, which are members of 19 functionally distinct protein complexes. We identified four proteins shared between complexes of different functionality thus charting segments of a protein interaction network. Concordance with the results of genome-wide two-hybrid screening was poor (14% of identified interactors overlapped) suggesting that the two approaches may provide complementary views on physical interactions within the proteome. PMID- 12096121 TI - A proteomic approach for identification of secreted proteins during the differentiation of 3T3-L1 preadipocytes to adipocytes. AB - We have undertaken a systematic proteomic approach to purify and identify secreted factors that are differentially expressed in preadipocytes versus adipocytes. Using one-dimensional gel electrophoresis combined with nanoelectrospray tandem mass spectrometry, proteins that were specifically secreted by 3T3-L1 preadipocytes or adipocytes were identified. In addition to a number of previously reported molecules that are up- or down-regulated during this differentiation process (adipsin, adipocyte complement-related protein 30 kDa, complement C3, and fibronectin), we identified four secreted molecules that have not been shown previously to be expressed differentially during the process of adipogenesis. Pigment epithelium-derived factor, a soluble molecule with potent antiangiogenic properties, was found to be highly secreted by preadipocytes but not adipocytes. Conversely, we found hippocampal cholinergic neurostimulating peptide, neutrophil gelatinase-associated lipocalin, and haptoglobin to be expressed highly by mature adipocytes. We also used liquid chromatography-based separation followed by automated tandem mass spectrometry to identify proteins secreted by mature adipocytes. Several additional secreted proteins including resistin, secreted acidic cysteine-rich glycoprotein/osteonectin, stromal cell-derived factor-1, cystatin C, gelsolin, and matrix metalloprotease-2 were identified by this method. To our knowledge, this is the first study to identify several novel secreted proteins by adipocytes by a proteomic approach using mass spectrometry. PMID- 12096122 TI - Proteomic analysis of shrimp white spot syndrome viral proteins and characterization of a novel envelope protein VP466. AB - White spot syndrome virus (WSSV) is at present one of the major pathogens in shrimp culture worldwide. The complete genome of this virus has been sequenced recently. To identify the structural and functional proteins of WSSV, the purified virions were separated by SDS-PAGE. Twenty-four protein bands were excised, in-gel digested with trypsin, and subjected to matrix-assisted laser desorption ionization-time of flight mass spectrometry and electrospray ionization tandem mass spectrometry, respectively. Eighteen proteins matching the open reading frames of WSSV genome were identified. Except for three known structural proteins and collagen, the functions of the remaining 14 proteins were unknown. Temporal analysis revealed that all the genes were transcribed in the late stage of WSSV infection except for vp121. Of the newly identified proteins, VP466 (derived from band 16) was further characterized. The cDNA encoding VP466 was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein. Specific antibody was generated with the purified GST-VP466 fusion protein. Western blot showed that the mouse anti-GST-VP466 antibody bound specifically to a 51-kDa protein of WSSV. Immunogold labeling revealed that VP466 protein is a component of the viral envelope. Results in this investigation thus proved the effectiveness of proteomic approaches for discovering new proteins of WSSV. PMID- 12096123 TI - Systematic identification of the genes affecting glycogen storage in the yeast Saccharomyces cerevisiae: implication of the vacuole as a determinant of glycogen level. AB - At the onset of nutrient limitation, the yeast Saccharomyces cerevisiae synthesizes glycogen to serve as a carbon and energy reserve. We undertook a systematic survey for the genes that affect glycogen accumulation by taking advantage of the strain deletion set generated by the Saccharomyces Genome Deletion Project. The strain collection analyzed contained some 4600 diploid homozygous null deletants, representing approximately 88% of all viable haploid disruptants. We identified 324 strains with low and 242 with elevated glycogen stores, accounting for 12.4% of the genes analyzed. The screen was validated by the identification of many of the genes known already to influence glycogen accumulation. Many of the mutants could be placed into coherent families. For example, 195 or 60% of the hypoaccumulators carry mutations linked to respiratory function, a class of mutants well known to be defective in glycogen storage. The second largest group consists of approximately 60 genes involved in vesicular trafficking and vacuolar function, including genes encoding 13 of 17 proteins involved in the structure or assembly of the vacuolar ATPase. These data are consistent with our recent findings that the process of autophagy has a significant impact on glycogen storage (Wang, Z., Wilson, W. A., Fujino, M. A., and Roach, P. J. (2001) Antagonistic controls of autophagy and glycogen accumulation by Snf1p, the yeast homolog of AMP-activated protein kinase, and the cyclin-dependent kinase Pho85p. Mol. Cell. Biol. 21, 5742-5752). Autophagy delivers glycogen to the vacuole, and we propose that the impaired vacuolar function associated with ATPase mutants (vma10 or vma22) results in reduced degradation and subsequent hyperaccumulation of glycogen. PMID- 12096124 TI - Analysis of mammalian peroxin interactions using a non-transcription-based bacterial two-hybrid assay. AB - In recent years, substantial progress has been made in the identification of proteins involved in peroxisome biogenesis. However, with the exception of the peroxisome-targeting signal receptors and the receptor docking proteins, the function of most of these proteins, called peroxins, remains largely unknown. One step toward elucidating the function of a protein is to identify its interacting partners. We have used a non-transcription-based bacterial two-hybrid system to analyze the interactions among a set of 12 mammalian peroxins and a yeast protein three-hybrid system to investigate whether proteins that interact with the same peroxin and have overlapping binding sites cooperate or compete for this site. Here we report a detailed interaction map of these peroxins and demonstrate that (i) farnesylation, and not the CAAX motif, of Pex19p strongly enhances its affinity for Pex13p; (ii) the CAAXmotif, and not farnesylation, of Pex19p strongly enhances its affinity for Pex11pbeta; and (iii) the C(3)HC(4) RING (really interesting new gene) finger domain of Pex12p does not alter the binding properties of Pex5p for the C-terminal peroxisome-targeting signal PTS1. Finally, we show that the Pex5p-Pex13p interaction is bridged by Pex14p and that the latter molecule exists predominantly as a dimer in vivo. Collectively, as demonstrated in this work with peroxins, these results indicate that the bacterial two-hybrid system is an attractive complementary approach to the conventional transcription-based yeast two-hybrid system for studying protein protein interactions. PMID- 12096125 TI - Isolation of protein subpopulations undergoing protein-protein interactions. AB - A new method is described for isolating and identifying proteins participating in protein-protein interactions in a complex mixture. The method uses a cyanogen bromide-activated Sepharose matrix to isolate proteins that are non-covalently bound to other proteins. Because the proteins are accessible to chemical manipulation, mass spectrometric identification of the proteins can yield information on specific classes of interacting proteins, such as calcium dependent or substrate-dependent protein interactions. This permits selection of a subpopulation of proteins from a complex mixture on the basis of specified interaction criteria. The new method has the advantage of screening the entire proteome simultaneously, unlike the two-hybrid system or phage display, which can only detect proteins binding to a single bait protein at a time. The method was tested by selecting rat brain extract for proteins exhibiting calcium-dependent protein interactions. Of 12 proteins identified by mass spectrometry, eight were either known calcium-binding proteins or proteins with known calcium-dependent protein interactions, indicating that the method is capable of enriching a subpopulation of proteins from a complex mixture on the basis of a specific class of protein interactions. Because only naturally occurring interactions of proteins in their native state are observed, this method will have wide applicability to studies of protein interactions in tissue samples and autopsy specimens, for screening for perturbations of protein-protein interactions by signaling molecules, pharmacological agents or toxins, and screening for differences between cancerous and untransformed cells. PMID- 12096126 TI - Evaluation of two-dimensional differential gel electrophoresis for proteomic expression analysis of a model breast cancer cell system. AB - The technique of fluorescent two-dimensional (2D) difference gel electrophoresis for differential protein expression analysis has been evaluated using a model breast cancer cell system of ErbB-2 overexpression. Labeling of paired cell lysate samples with N-hydroxy succinimidyl ester-derivatives of fluorescent Cy3 and Cy5 dyes for separation on the same 2D gel enabled quantitative, sensitive, and reproducible differential expression analysis of the cell lines. SyproRuby staining was shown to be a highly sensitive and 2D difference gel electrophoresis compatible method for post-electrophoretic visualization of proteins, which could then be picked and identified by matrix-assisted laser-desorption ionization mass spectroscopy. Indeed, from these experiments, we have identified multiple proteins that are likely to be involved in ErbB-2-mediated transformation. A triple dye labeling methodology was used to identify proteins differentially expressed in the cell system over a time course of growth factor stimulation. A Cy2-labeled pool of samples was used as a standard with all Cy3- and Cy5-labeled sample pairs to facilitate cross-gel quantitative analysis. DeCyder (Amersham Biosciences, Inc.) software was used to distinguish clear statistical differences in protein expression over time and between the cell lines. PMID- 12096127 TI - Combinatorial selection of RNA ligands for complex cellular targets : the RNA liagands-based proteomics. AB - This study explores the selection of high affinity RNA ligands for the complex cellular targets present in crude HeLa nuclear extract through directed evolution and deconvolution. RNA ligands for the mixed nuclear targets were selected from around 6 x 10(14) RNA sequences through an iterated enrichment process. RNA ligands for various gene products of the extract were simultaneously selected and were shown to specifically interact with their target molecules. The target molecules were isolated from the nuclear extract by affinity chromatography using columns tagged with the RNA ligands, resolved on two-dimensional gels, and identified by mass spectrometry. These RNA ligands may be useful in characterizing novel functions of cellular proteins and modulating complex molecular events. PMID- 12096128 TI - Describing biological protein interactions in terms of protein states and state transitions: the LiveDIP database. AB - Biological protein-protein interactions differ from the more general class of physical interactions; in a biological interaction, both proteins must be in their proper states (e.g. covalently modified state, conformational state, cellular location state, etc.). Also in every biological interaction, one or both interacting molecules undergo a transition to a new state. This regulation of protein states through protein-protein interactions underlies many dynamic biological processes inside cells. Therefore, understanding biological interactions requires information on protein states. Toward this goal, DIP (the Database of Interacting Proteins) has been expanded to LiveDIP, which describes protein interactions by protein states and state transitions. This additional level of characterization permits a more complete picture of the protein-protein interaction networks and is crucial to an integrated understanding of genome scale biology. The search tools provided by LiveDIP, Pathfinder, and Batch Search allow users to assemble biological pathways from all the protein-protein interactions collated from the scientific literature in LiveDIP. Tools have also been developed to integrate the protein-protein interaction networks of LiveDIP with large scale genomic data such as microarray data. An example of these tools applied to analyzing the pheromone response pathway in yeast suggests that the pathway functions in the context of a complex protein-protein interaction network. Seven of the eleven proteins involved in signal transduction are under negative or positive regulation of up to five other proteins through biological protein-protein interactions. During pheromone response, the mRNA expression levels of these signaling proteins exhibit different time course profiles. There is no simple correlation between changes in transcription levels and the signal intensity. This points to the importance of proteomic studies to understand how cells modulate and integrate signals. Integrating large scale, yeast two-hybrid data with mRNA expression data suggests biological interactions that may participate in pheromone response. These examples illustrate how LiveDIP provides data and tools for biological pathway discovery and pathway analysis. PMID- 12096129 TI - 2D differential in-gel electrophoresis for the identification of esophageal scans cell cancer-specific protein markers. AB - The reproducibility of conventional two-dimensional (2D) gel electrophoresis can be improved using differential in-gel electrophoresis (DIGE), a new emerging technology for proteomic analysis. In DIGE, two pools of proteins are labeled with 1-(5-carboxypentyl)-1'-propylindocarbocyanine halide (Cy3) N-hydroxy succinimidyl ester and 1-(5-carboxypentyl)-1'-methylindodi-carbocyanine halide (Cy5) N-hydroxysuccinimidyl ester fluorescent dyes, respectively. The labeled proteins are mixed and separated in the same 2D gel. 2D DIGE was applied to quantify the differences in protein expression between laser capture microdissection-procured esophageal carcinoma cells and normal epithelial cells and to define cancer-specific and normal-specific protein markers. Analysis of the 2D images from protein lysates of approximately 250,000 cancer cells and normal cells identified 1038 protein spots in cancer cell lysates and 1088 protein spots in normal cell lysates. Of the detected proteins, 58 spots were up regulated by >3-fold and 107 were down-regulated by >3-fold in cancer cells. In addition to previously identified down-regulated protein annexin I, tumor rejection antigen (gp96) was found up-regulated in esophageal squamous cell cancer. Global quantification of protein expression between laser capture microdissected patient-matched cancer cells and normal cells using 2D DIGE in combination with mass spectrometry is a powerful tool for the molecular characterization of cancer progression and identification of cancer-specific protein markers. PMID- 12096130 TI - Increased frequency of cysteine, tyrosine, and phenylalanine residues since the last universal ancestor. AB - Analysis of extant proteomes has the potential of revealing how amino acid frequencies within proteins have evolved over biological time. Evidence is presented here that cysteine, tyrosine, and phenylalanine residues have substantially increased in frequency since the three primary lineages diverged more than three billion years ago. This inference was derived from a comparison of amino acid frequencies within conserved and non-conserved residues of a set of proteins dating to the last universal ancestor in the face of empirical knowledge of the relative mutability of these amino acids. The under-representation of these amino acids within last universal ancestor proteins relative to their modern descendants suggests their late introduction into the genetic code. Thus, it appears that extant ancient proteins contain evidence pertaining to early events in the formation of biological systems. PMID- 12096131 TI - Hydrogen exchange-mass spectrometry: optimization of digestion conditions. AB - The direct linkage between folded structures of proteins and their function has increased the need for high resolution structures. In addition, there is a need for analytical methods for detecting and locating changes in the folded structures of proteins under a wide variety of conditions. The rates at which hydrogens located at peptide amide linkages undergo isotopic exchange has become the basis for an important method for detecting such structural changes. When detected by mass spectrometry, hydrogen exchange can be used to study dilute solutions of large proteins and protein complexes with very high sensitivity. To locate structural changes, labeled proteins are often digested with acid proteases to form peptides whose hydrogen/deuterium levels are determined by mass spectrometry. This approach is successful only when the protein can be digested rapidly under conditions where isotope exchange is slow. This study describes how columns packed with immobilized pepsin can be used to reduce the digestion time and to provide an effective means for separating the pepsin from the isotopically labeled fragments. These columns are part of an on-line system that facilitates both rapid digestion of low concentrations of protein and concentration of the peptides. PMID- 12096132 TI - Getting more from less: algorithms for rapid protein identification with multiple short peptide sequences. AB - We describe two novel sequence similarity search algorithms, FASTS and FASTF, that use multiple short peptide sequences to identify homologous sequences in protein or DNA databases. FASTS searches with peptide sequences of unknown order, as obtained by mass spectrometry-based sequencing, evaluating all possible arrangements of the peptides. FASTF searches with mixed peptide sequences, as generated by Edman sequencing of unseparated mixtures of peptides. FASTF deconvolutes the mixture, using a greedy heuristic that allows rapid identification of high scoring alignments while reducing the total number of explored alternatives. Both algorithms use the heuristic FASTA comparison strategy to accelerate the search but use alignment probability, rather than similarity score, as the criterion for alignment optimality. Statistical estimates are calculated using an empirical correction to a theoretical probability. These calculated estimates were accurate within a factor of 10 for FASTS and 1000 for FASTF on our test dataset. FASTS requires only 15-20 total residues in three or four peptides to robustly identify homologues sharing 50% or greater protein sequence identity. FASTF requires about 25% more sequence data than FASTS for equivalent sensitivity, but additional sequence data are usually available from mixed Edman experiments. Thus, both algorithms can identify homologues that diverged 100 to 500 million years ago, allowing proteomic identification from organisms whose genomes have not been sequenced. PMID- 12096133 TI - Selective detection of membrane proteins without antibodies: a mass spectrometric version of the Western blot. AB - A method has been developed, called the mass western experiment in analogy to the Western blot, to detect the presence of specific proteins in complex mixtures without the need for antibodies. Proteins are identified with high sensitivity and selectivity, and their abundances are compared between samples. Membrane protein extracts were labeled with custom isotope-coded affinity tag reagents and digested, and the labeled peptides were analyzed by liquid chromatography-tandem mass spectrometry. Ions corresponding to anticipated tryptic peptides from the proteins of interest were continuously subjected to collision-induced dissociation in an ion trap mass spectrometer; heavy and light isotope-coded affinity tag-labeled peptides were simultaneously trapped and fragmented accomplishing identification and quantitation in a single mass spectrum. This application of ion trap selective reaction monitoring maximizes sensitivity, enabling analysis of peptides that would otherwise go undetected. The cell surface proteins prostate stem cell antigen (PSCA) and ErbB2 were detected in prostate and breast tumor cell lines in which they are expressed in known abundances spanning orders of magnitude. PMID- 12096134 TI - Application of microfluidic devices to proteomics research: identification of trace-level protein digests and affinity capture of target peptides. AB - This report describes an integrated and modular microsystem providing rapid analyses of trace-level tryptic digests for proteomics applications. This microsystem includes an autosampler, a microfabricated device comprising a large channel (2.4 microl total volume), an array of separation channels, together with a low dead volume enabling the interface to nanoelectrospray mass spectrometry. The large channel of this microfluidic device provides a convenient platform to integrate C(18) reverse phase packing or other type of affinity media such as immobilized antibodies or immobilized metal affinity chromatography beads thus enabling affinity selection of target peptides prior to electrophoretic separation and mass spectrometry analyses on a quadrupole/time-of-flight instrument. Sequential injection, preconcentration, and separation of peptide standards and tryptic digests are achieved with a throughput of up to 12 samples/per h and a concentration detection limit of approximately 5 nM (25 fmol on chip). Replicate injections of peptide mixtures indicated that reproducibility of migration time was 1.2-1.8%, whereas relative standard deviation ranging from 9.2 to 11.8% are observed on peak heights. The application of this device for trace-level protein identification is demonstrated for two-dimensional gel spots obtained from extracts of human prostatic cancer cells (LNCap) using both peptide mass-fingerprint data base searching and on-line tandem mass spectrometry. Enrichment of target peptides prior to mass spectral analyses is achieved using c myc-specific antibodies immobilized on protein G-Sepharose beads and facilitates the identification of antigenic peptides spiked at a level of 20 ng/ml in human plasma. Affinity selection is also demonstrated for gel-isolated protein bands where tryptic phosphopeptides are captured on immobilized metal affinity chromatography beads and subsequently separated and characterized on this microfluidic system. PMID- 12096135 TI - Charting the protein complexome in yeast by mass spectrometry. AB - It has become evident over the past few years that many complex cellular processes, including control of the cell cycle and ubiquitin-dependent proteolysis, are carried out by sophisticated multisubunit protein machines that are dynamic in abundance, post-translational modification state, and composition. To understand better the nature of the macromolecular assemblages that carry out the cell cycle and ubiquitin-dependent proteolysis, we have used mass spectrometry extensively over the past few years to characterize both the composition of various protein complexes and the modification states of their subunits. In this article we review some of our recent efforts, and describe a promising new approach for using mass spectrometry to dissect protein interaction networks. PMID- 12096136 TI - C-mannosylation and o-fucosylation of thrombospondin type 1 repeats. AB - The final chemical structure of a newly synthesized protein is often only attained after further covalent modification. Ideally, a comprehensive proteome analysis includes this aspect, a task that is complicated by our incomplete knowledge of the range of possible modifications and often by the lack of suitable analysis methods. Here we present two recently discovered, unusual forms of protein glycosylation, i.e. C-mannosylation and O-fucosylation. Their analysis by a combined mass spectrometric approach is illustrated with peptides from the thrombospondin type 1 repeats (TSRs) of the recombinant axonal guidance protein F spondin. Nano-electrospray ionization tandem-mass spectrometry of isolated peptides showed that eight of ten Trp residues in the TSRs of F-spondin are C mannosylated. O-Fucosylation sites were determined by a recently established nano electrospray ionization quadrupole time-of-flight tandem-mass spectrometry approach. Four of five TSRs carry the disaccharide Hex-dHex-O-Ser/Thr in close proximity to the C-mannosylation sites. In analogy to thrombospondin-1, we assume this to be Glc-Fuc-O-Ser/Thr. Our current knowledge of these glycosylations will be discussed. PMID- 12096137 TI - Quantitative protein profiling using two-dimensional gel electrophoresis, isotope coded affinity tag labeling, and mass spectrometry. AB - Quantitative protein profiling is an essential part of proteomics and requires new technologies that accurately, reproducibly, and comprehensively identify and quantify the proteins contained in biological samples. We describe a new strategy for quantitative protein profiling that is based on the separation of proteins labeled with isotope-coded affinity tag reagents by two-dimensional gel electrophoresis and their identification and quantification by mass spectrometry. The method is based on the observation that proteins labeled with isotopically different isotope-coded affinity tag reagents precisely co-migrate during two dimensional gel electrophoresis and that therefore two or more isotopically encoded samples can be separated concurrently in the same gel. By analyzing changes in the proteome of yeast (Saccharomyces cerevisiae) induced by a metabolic shift we show that this simple method accurately quantifies changes in protein abundance even in cases in which multiple proteins migrate to the same gel coordinates. The method is particularly useful for the quantitative analysis and structural characterization of differentially processed or post translationally modified forms of a protein and is therefore expected to find wide application in proteomics research. PMID- 12096138 TI - A proteomic approach for the identification of cell-surface proteins shed by metalloproteases. AB - Proteolytic cleavage (shedding) of extracellular domains of many membrane proteins by metalloproteases is an important regulatory mechanism used by mammalian cells in response to environmental and physiological changes. Here we describe a proteomic system for analyzing cell surface shedding. The method utilized short-term culture supernatants from induced cells as starting material, followed by lectin-affinity purification, deglycosylation, and polyacrylamide gel electrophoresis separation. Relative quantitation of proteins was achieved via isotope dilution. In this study, a number of proteins already known to be shed were identified from activated monocytes and endothelial cells, thereby validating the method. In addition, a group of proteins were newly identified as being shed. The method provides an unbiased means to screen for shed proteins. PMID- 12096139 TI - Genome-wide study of gene copy numbers, transcripts, and protein levels in pairs of non-invasive and invasive human transitional cell carcinomas. AB - Gain and loss of chromosomal material is characteristic of bladder cancer, as well as malignant transformation in general. The consequences of these changes at both the transcription and translation levels is at present unknown partly because of technical limitations. Here we have attempted to address this question in pairs of non-invasive and invasive human bladder tumors using a combination of technology that included comparative genomic hybridization, high density oligonucleotide array-based monitoring of transcript levels (5600 genes), and high resolution two-dimensional gel electrophoresis. The results showed that there is a gene dosage effect that in some cases superimposes on other regulatory mechanisms. This effect depended (p < 0.015) on the magnitude of the comparative genomic hybridization change. In general (18 of 23 cases), chromosomal areas with more than 2-fold gain of DNA showed a corresponding increase in mRNA transcripts. Areas with loss of DNA, on the other hand, showed either reduced or unaltered transcript levels. Because most proteins resolved by two-dimensional gels are unknown it was only possible to compare mRNA and protein alterations in relatively few cases of well focused abundant proteins. With few exceptions we found a good correlation (p < 0.005) between transcript alterations and protein levels. The implications, as well as limitations, of the approach are discussed. PMID- 12096140 TI - The chloroplast grana proteome defined by intact mass measurements from liquid chromatography mass spectrometry. AB - Proteomics seeks to address the entire complement of protein gene products of an organism, but experimental analysis of such complex mixtures is biased against low abundance and membrane proteins. Electrospray-ionization mass spectrometry coupled with reverse-phase chromatography was used to separate and catalogue all detectable proteins in samples of photosystem II-enriched thylakoid membrane subdomains (grana) from pea and spinach. Around 90 intact mass tags were detected corresponding to approximately 40 gene products with variable post-translational covalent modifications. Provisional identity of 30 of these gene products was proposed based upon coincidence of measured mass with that calculated from genomic sequence. Analysis of isolated photosystem II complexes allowed detection and resolution of a minor population of D1 (PsbA) that was apparently palmitoylated and not detected in less purified preparations. Based upon observed +80-Da adducts, D1, D2 (PsbD), CP43 (PsbC), two Lhcbs, and PsbH were confirmed to be phosphorylated, and a new phosphoprotein was proposed to be the product of psbT. The appearance of a second +80-Da adduct on PsbH provides direct evidence for a second phosphorylation site on PsbH, complicating interpretation of its role in regulation of thylakoid membrane organization and function, including light-state transitions. Adducts of +32 Da, presumably arising from oxidative modification during illumination, were associated with more highly phosphorylated forms of PsbH implying a relationship between the two phenomena. Intact mass proteomics of organellar subfractions and more highly purified protein complexes provides increasingly detailed insights into functional genomics of photosynthetic membranes. PMID- 12096141 TI - Chemical approaches for functionally probing the proteome. AB - With the availability of complete genome sequences, emphasis has shifted toward the understanding of protein function. We have developed a functional proteomic methodology that makes use of chemically reactive fluorescent probes to profile and identify enzymes in complex mixtures by virtue of their catalytic activity. This methodology allows a comparison of changes in activity of multiple enzymes under a variety of conditions using a single two-dimensional separation. The probes can also be used to localize active enzymes in intact cells using fluorescence microscopy. Furthermore, the probes enable screens for selective small molecule inhibitors of each enzyme family member within crude lysates or intact cells. Ultimately, this technology allows the rapid identification of potential drug targets and small molecule lead compounds targeted to them. PMID- 12096142 TI - Fully automated two-dimensional capillary electrophoresis for high sensitivity protein analysis. AB - We report a system for automated protein analysis. In the system, proteins are labeled with the fluorogenic reagent 3-(2-furoyl)quinoline-2-carboxaldehyde, which reacts with lysine residues and creates a highly fluorescent product. These labeled proteins are analyzed by submicellar capillary electrophoresis at pH 7.5 to perform a first dimension separation. Once the first components migrate from the capillary, a fraction is transferred to a second dimension capillary, where electrophoresis is performed at pH 11.1 to further separate the proteins. Laser induced fluorescence is used as an ultrasensitive detector of the separated proteins. Successive fractions are transferred from the first dimension capillary to the second dimension capillary for further separation to generate, in serial fashion, a two-dimensional electropherogram. The transfer of fractions is computer-controlled; there is no operator intervention once the sample has been injected. Zeptomoles of labeled proteins are detected, providing exquisite sensitivity. PMID- 12096143 TI - Vascular proteomics and subtractive antibody expression cloning. AB - The cloning of genes expressing proteins that are differentially expressed in the organ microvasculature has the potential to address a variety of problems ranging from the analysis of disease pathogenesis to drug targeting for particular tissues. This study describes a methodology designed to analyze differential protein expression in the brain microvasculature. The method can be applied to other organs and is particularly suited to the cloning of cDNAs encoding membrane proteins. The technology merges a tissue-specific polyclonal antiserum with a cDNA library expression cloning system. The tissue-specific antiserum is subtracted with protein extracts from control tissues to remove those antibodies that recognize common antigenic proteins. Then, the depleted antiserum is used to expression clone tissue-specific proteins from a cDNA library expressed in mammalian cells. The methodology was evaluated with a rabbit polyclonal antiserum prepared against purified bovine brain capillaries. The antiserum was absorbed with acetone powders of liver and kidney and then used to screen a bovine brain capillary cDNA library in COS cells. The initial clone detected with this expression methodology was the Lutheran membrane glycoprotein, which is specifically expressed at the brain microvasculature compared with liver and kidney tissues. This subtractive expression cloning methodology provides a new approach to "vascular proteomics" and to the detection of proteins specifically expressed at the microvasculature, including membrane proteins. PMID- 12096145 TI - What do Cochrane reviews tell us about anti-thrombotic therapy in heart failure with sinus rhythm? PMID- 12096146 TI - Is exercise blood pressure a marker of vascular endothelial function? PMID- 12096148 TI - Erythrocyte thiopurine methyl transferase assessment prior to azathioprine use in the UK. AB - BACKGROUND: Individuals with low activity of a key metabolic enzyme, thiopurine methyl transferase (TPMT), are more susceptible to azathioprine-induced myelosuppression. AIM: To determine the pattern of use of TPMT activity estimation, with respect to azathioprine use, by medical practitioners in the UK. DESIGN: Retrospective analysis of assay use. METHODS: We analysed all test results (n=3291), and patient and practitioner details, from inception of TPMT assay in 1990 to the end of December 2000, held at the Purine Research Laboratory, Guy's Hospital, London. Patient details were anonymized. Repeat analyses and requests from outside the UK were excluded. RESULTS: The male:female ratio was approximately equal and the mean age was 46.6 (range 0.5-97) years. Thirteen different medical specialities requested assays; Dermatology and Gastroenterology were the most frequent users, together accounting for 86% of requests. The numbers of centres requesting the assay varied widely both within and between different specialities. Some 80% of individuals had normal TPMT activity, 9% enzymic activity above normal, and 10% low activity. Fifteen had no detectable enzymic activity: 0.45% (1:220) of the study population. DISCUSSION: This incidence of undetectable enzyme activity is significantly higher than the previously reported level of 1:300 derived from smaller studies, and makes the economics of screening more attractive. PMID- 12096147 TI - Hyponatraemic states following 3,4-methylenedioxymethamphetamine (MDMA, 'ecstasy') ingestion. AB - BACKGROUND: Life-threatening and fatal hyponatraemic complications following ecstasy use have previously been documented. AIM: To define clinical features of hyponatraemia following the ingestion of 3,4-methylenedioxymethamphetamine (MDMA, 'ecstasy'). DESIGN: Retrospective case series. METHODS: All enquiries to the London centre of the National Poisons Information Service (NPIS) between December 1993 and March 1996 were screened for cases of MDMA use associated with hyponatraemia (serum sodium <130 mmol/l). History of fluid consumption, presenting features and subsequent clinical course were recorded. RESULTS: Seventeen patients, aged 15-26 years, were identified. Serum sodium levels ranged between 107 mmol/l and 128 mmol/l. In six patients, biochemical results were consistent with inappropriate secretion of antidiuretic hormone (SIADH). Analytical confirmation of MDMA ingestion was obtained in 10 patients. Ten patients were known to have ingested a large amount of non-alcoholic or alcoholic fluid. The clinical pattern was remarkably uniform, with initial vomiting and disturbed behaviour, followed in 11 patients by seizures. Drowsiness, a mute state and disorientation were observed for up to 3 days. Two patients died; 14 made a complete recovery. DISCUSSION: MDMA can cause life-threatening hyponatraemic encephalopathy when accompanied by excessive fluid ingestion. The mechanism involves inappropriate secretion of antidiuretic hormone. PMID- 12096149 TI - A community study of health promotion in rural West Africa: details of a household survey and population census. AB - BACKGROUND: Hypertension is an important problem in sub-Saharan Africa. The low use of processed food in this area makes a population approach to reducing salt intake feasible. AIM: To create an age-sex register for 12 villages in Ghana as the first stage of a community study of the effect of dietary salt reduction on blood pressure and urinary sodium excretion in West African villagers. DESIGN: Household survey and population census. METHODS: Over three months, village maps were sketched and a complete list of total number of households, adults (with age and gender) and children in each village was obtained. RESULTS: The six semi urban villages were larger than the six rural villages (10368 vs. 6597 inhabitants) and almost half the total population was under 16. CONCLUSIONS: Accurate census data are important in the design, implementation and interpretation of community studies and intervention trials. We outline the methods by which census data can be collected in rural and semi-urban sub-Saharan African villages, and emphasize the importance of painstaking, thorough work in the collection of such data. PMID- 12096150 TI - Anticoagulation for heart failure in sinus rhythm: a Cochrane systematic review. AB - BACKGROUND: Patients with chronic heart failure (heart failure) are at risk of thromboembolic events, and coronary ischaemic events also contribute to the progression of heart failure. Long-term oral anticoagulation is established in certain groups, including patients with heart failure and atrial fibrillation, but there is wide variation in the use of oral anticoagulation in the broader heart failure population. OBJECTIVE: To determine whether long-term oral anticoagulation reduces total deaths and/or major thromboembolic events in patients with heart failure. DESIGN: Systematic review. DATA SOURCES: Reference lists of papers resulting from this search, electronic database searching (MEDLINE, EMBASE, DARE), and abstracts from national and international cardiovascular meetings were studied to identify unpublished studies. Relevant authors of these studies were contacted to obtain further data. SELECTION CRITERIA: Randomized controlled trials (RCTs) comparing oral anticoagulants with control or placebo. Non-randomized studies were included, as they may help in assessing side-effects. Other inclusion criteria included duration of treatment > or =1 month, and adults with heart failure due to any underlying cause. Inclusion decisions were duplicated, and disagreement resolved by discussion or a third party. RESULTS: One recent pilot RCT compared warfarin, aspirin and no antithrombotic therapy, but no definitive data have yet been published. Three small prospective studies of warfarin in heart failure were also identified, but were over 50 years old, with methods considered unreliable today: in these, anticoagulation was more efficacious than control in reducing all-cause death (OR 0.64; 95%CI 0.45-0.90) and cardiovascular events (OR 0.26; 95%CI 0.16-0.43). Four retrospective non-randomized cohort analyses and three small observational studies of oral anticoagulation in heart failure included differing populations of heart failure patients, and reported contradictory results. CONCLUSIONS: Limited evidence from randomized trials and observational studies found a reduction in mortality and cardiovascular events with anticoagulants compared to controls. This evidence should be interpreted with caution. Although oral anticoagulation is indicated in certain groups of patients with heart failure (e.g. atrial fibrillation), the available data do not support its routine use in heart failure patients who remain in sinus rhythm. PMID- 12096151 TI - Antiplatelet agents versus control or anticoagulation for heart failure in sinus rhythm: a Cochrane systematic review. AB - BACKGROUND: Heart failure predisposes to stroke and thromboembolism, which in turn contribute to the high mortality and morbidity in heart failure. OBJECTIVES: To determine the effect of antiplatelet agents, compared to placebo or anticoagulant therapy, on death and/or major thromboembolic events in adults with heart failure who are in sinus rhythm. DESIGN: Systematic review of randomized parallel group placebo or controlled trials comparing oral antiplatelet therapy with control or anticoagulation therapy in adults with chronic heart failure in sinus rhythm. DATA SOURCES: Reference lists of papers resulting from this search, electronic database searching (MEDLINE, EMBASE, DARE), and abstracts from national and international cardiovascular meetings were studied to identify unpublished studies. Relevant authors of these studies were contacted to obtain further data. SELECTION CRITERIA: These included duration of treatment of at least 1 month, and adults with heart failure due to any underlying cause. To assess any adverse effects, cohort study and non-randomized controlled studies were assessed. Inclusion decisions were duplicated, disagreement resolved by discussion or a third party. No meta-analyses were performed, as no data were available from randomized comparisons. RESULTS: One randomized controlled trial of warfarin vs. aspirin vs. no antithrombotic therapy was found, but no definitive data have yet been published. Three retrospective, non-randomized cohort studies from large trials examining the role of ACE inhibitors have examined the role of aspirin therapy with and without anticoagulant therapy in patients with heart failure and/or left ventricular systolic dysfunction were identified, but the results from these trials were conflicting. A possible interaction with ACE inhibitors may reduce the efficacy of aspirin, although this evidence is from retrospective analyses of trial cohorts. CONCLUSIONS: At present there is no evidence from long term RCTs to recommend use of aspirin to prevent thromboembolism in patients with heart failure in sinus rhythm. There is also no evidence to indicate superior effects from oral anticoagulation, when compared to aspirin, in patients with heart failure in sinus rhythm. PMID- 12096152 TI - Towards equity in global health knowledge. PMID- 12096153 TI - Acute hyponatraemia and 'ecstasy': insights from a quantitative and integrative analysis. AB - A 20-year-old woman attended a 'rave party' where she took the drug 3,4 methylenedioxymethamphetamine (MDMA, 'ecstasy'). She had used this drug previously without serious adverse effects. On this occasion, while both she and her friends drank a large quantity of water, only she became seriously ill. The initial manifestation was an altered sensorium; several hours later she had a grand mal seizure. In the Emergency Department, the most striking features were the severe degree of hyponatraemia (112 mmol/l) and cerebral oedema. To explain the basis for this life-threatening clinical presentation, an imaginary consultation was sought with Professor McCance. Using both a deductive and a quantitative analysis that involved several medical subspecialties, he illustrated that a simple story of water ingestion and vasopressin release was not sufficient to explain her hyponatraemia. It was only after events in her gastrointestinal tract were analysed that a plausible hypothesis could be constructed. PMID- 12096154 TI - Flies in the colon. PMID- 12096156 TI - Responding to tragedy. PMID- 12096158 TI - The U.S Supreme Court narrows the definition of disability under the Americans With Disabilities Act. PMID- 12096159 TI - Improving social functioning in schizophrenia by playing the train game. AB - Social impairments have long been recognized as a core feature of schizophrenia. Poor social, self-care, and vocational functioning are criteria for a diagnosis of schizophrenia in most diagnostic systems. Consequently, improving the social behaviors of persons with schizophrenia has been a key target of psychiatric rehabilitation techniques. One such technique, social skills training, has demonstrated effectiveness in yielding skill acquisition, durability, and generalization (1) and has been recognized as a psychosocial treatment of choice for schizophrenia (2). Nevertheless, a number of limitations to the benefits that may be achieved through skills training have been described. For example, the cognitive impairments of persons with schizophrenia, such as poor sustained attention and verbal memory deficits, have been shown to limit the acquisition of social skills (3). Moreover, one study showed that patients with negative symptoms, such as apathy, anhedonia, and amotivation, have an impaired capacity to benefit from social skills training (4). However, the results of a more recent study suggest that only persons with primary, deficit-type negative symptoms-not those with negative symptoms secondary to positive psychotic symptoms, depression, or extrapyramidal side effects-are unable to benefit from conventional social skills training (5). Recognizing these obstacles to implementing a social skills training program for persons with schizophrenia, Torres and his colleagues have created a board game called El Tren (The Train). Several characteristics of El Tren make it ideal for use with this population. First, the game is behaviorally oriented and has an emphasis on positive reinforcement and shaping. Second, it is sensitive to the cognitive limitations of the participants, emphasizing repetition and procedural learning. Third, it is designed to overcome participants' negative symptoms by being entertaining and fun. In this month's column, these authors describe El Tren and present the results of a randomized controlled study of its efficacy. PMID- 12096160 TI - Medicaid managed care and substance abuse treatment. PMID- 12096161 TI - Strengthening the consumer voice in managed care: v. helping professionals listen. PMID- 12096162 TI - Current trends in the community treatment of seriously emotionally disturbed youths. PMID- 12096163 TI - Determinants of counseling for children in Manhattan after the September 11 attacks. AB - OBJECTIVE: After the September 11 terrorist attacks, many adults and children received counseling. The authors assessed the prevalence and correlates of counseling for experiences related to the disaster received by children aged four to 18 years living in Manhattan. METHODS: From a representative sample of 1,008 adult residents of Manhattan who were living below 110th Street five to eight weeks after the attacks, the authors interviewed 112 parents or primary caretakers about their child's level of exposure to the disaster, the extent of loss, receipt of counseling services, and behavioral reaction. RESULTS: Overall, 22 percent of the children had received some form of counseling related to their experiences after the disaster. More than half of the counseling received (58 percent) was delivered in schools. Predictors of counseling in a multivariate model were male sex (odds ratio=5.3), having a parent with current posttraumatic stress disorder related to the attacks (OR=4.3), and having at least one sibling living in the household (OR=3.6). CONCLUSIONS: Parents' own level of posttraumatic stress was associated with whether their children received counseling related to the September 11 attacks. This finding has important implications, because parents act as decision makers for their children in seeking health care. After the terrorist attacks, school and health care personnel provided early intervention counseling in Manhattan. PMID- 12096164 TI - The effect on diagnostic rates of assigning patients to ethnically focused inpatient psychiatric units. AB - OBJECTIVE: The authors examined whether assigning patients from three ethnic groups-blacks, Latinos, and Asians-to three psychiatric inpatient units that provided culturally appropriate treatment to those groups would affect rates of diagnosis of various psychiatric disorders. METHODS: Retrospective administrative data for 5,983 inpatients at a large urban community hospital with several ethnically focused units were examined. The data represented 10,645 admissions between 1989 and 1996. Chi square analyses and Stuart-Maxwell tests of symmetry and homogeneity were used to assess the relationship between matching patients to ethnically focused units and the rates of major psychiatric illnesses among Asian, black, and Latino patients compared with whites. RESULTS: Ethnic differences in diagnostic rates were consistent with the results of previous studies. Black patients had more diagnoses of psychotic disorders and fewer diagnoses of affective disorders than other ethnic minorities or whites, and Latino patients had more nonspecific diagnoses. Matching inpatients to ethnically focused units did not have a marked effect on patterns of diagnoses among black patients, but an association was observed for Latino patients, particularly those who had only one admission. No significant diagnostic differences were found between Asian patients and whites, irrespective of whether the Asian patients had been ethnically matched to a specialty focus unit. CONCLUSIONS: The effect of referring inpatients with serious mental illnesses to an ethnically focused psychiatric unit varied by ethnic group, probably because each specialty unit functioned differently, depending on the needs of its particular patient population. PMID- 12096165 TI - The effect on treatment outcomes of assigning patients to ethnically focused inpatient psychiatric units. AB - OBJECTIVE: The authors examined whether assigning patients from three ethnic groups-blacks, Latinos, and Asians-to three ethnically focused psychiatric inpatient units would affect treatment outcome. METHODS: Retrospective administrative data for 5,983 inpatients at a large urban community hospital with several ethnically focused units were examined. The data represented 10,645 admissions between 1989 and 1996. Cox proportional-hazards models, logistic and multinomial regressions, and chi square analyses were used to assess the relationship between matching patients to ethnically focused units and time to rehospitalization, referral destination on discharge, and length of stay for Asian, black, and Latino patients. RESULTS: Ethnic matching status was strongly associated with referral destination for Asian and Latino patients but not for black patients. Asian and Latino patients who had been treated on the appropriate ethnically focused units were more frequently sent to outpatient or residential treatment (71 to 73 percent of discharges) than unmatched patients, black patients, and white patients (44 to 49 percent of discharges), who more frequently refused follow-up or were sent to locked facilities. No association was found between matching status and time to rehospitalization or length of stay for any ethnic group. CONCLUSIONS: Matching inpatients to ethnically focused psychiatric units was related to referral destination at discharge. Matched patients were more likely than unmatched patients to accept referral to postdischarge treatment, which has been shown previously to reduce readmission rates. Among persons with serious mental illness, matching patients to ethnically focused units may be important for enhancing communication and trust as a means of improving participation in ongoing treatment programs. PMID- 12096167 TI - Body weight changes associated with psychopharmacology. AB - OBJECTIVE: The authors discuss changes in body weight associated with various psychopharmaceuticals. METHODS: A large number of articles and books about drug induced changes in body weight, selected on the basis of various literature searches and the authors' clinical experiences with psychopharmaceuticals, were reviewed. RESULTS: Many psychotropic drugs with antipsychotic, mood stabilizing, and antidepressant properties are associated with weight gain. Others, such as fluoxetine, isocarboxazid, nefazadone, topiramate, and psychostimulants, may cause weight loss. The antipsychotic drugs chlorpromazine, clozapine, and olanzapine are often associated with weight gain. Among antidepressants, amitriptyline and mirtazapine are known to cause weight gain. However, reductions are sometimes observed, and each antidepressant has its own unique weight-effect profile. Mood stabilizers, especially valproate-related products, are also associated with weight gain. CONCLUSIONS: Careful monitoring and consideration of alternative therapies are essential. PMID- 12096166 TI - Long-term use of VA mental health services by older patients with substance use disorders. AB - OBJECTIVE: The authors studied long-term patterns and predictors of use of mental health services by older surviving patients with substance use disorders in the Veterans Affairs (VA) health care system. METHODS: In this prospective longitudinal study, patient treatment records were used to determine long-term (ten-year) patterns and predictors of use of VA mental health services in a nationwide cohort of 10,678 surviving patients with a substance use disorder who were 55 years of age or older. The patients were categorized into three groups based on diagnosis during the index episode: patients with alcohol or drug abuse or dependence, patients with alcohol or drug psychosis, and patients with both a substance use and a psychiatric disorder. Most of the patients had alcohol use disorders. RESULTS: Over the ten-year period, successively fewer patients obtained outpatient and inpatient mental health care. Among patients who did obtain such care, the intensity of service use increased. Medical care did not substitute for mental health treatment. Younger age, being unmarried, and having a more severe disorder were associated with a greater likelihood of mental health service use over the ten-year period. Patients with a dual diagnosis were significantly more likely to obtain outpatient mental health care. Treatment on a residential care unit and longer initial hospital stay were associated with a lower rate of mental health readmissions. CONCLUSIONS: Of the substantial number of patients with substance use disorders who survive into old age, those with more long-standing substance use problems and with dual diagnoses have the greatest need for long-term mental health treatment. PMID- 12096168 TI - Nonpsychiatric illness among primary care patients with trauma histories and posttraumatic stress disorder. AB - OBJECTIVE: The authors examined the relationship between posttraumatic stress disorder (PTSD), trauma, and self-reported nonpsychiatric medical conditions in a sample of 502 primary care patients with one or more anxiety disorders. METHODS: Primary care patients with one or more DSM-IV anxiety disorders were assessed for comorbid psychiatric and substance use problems and for a history of trauma. These individuals also completed a self-report measure of current and lifetime medical conditions, lifetime tobacco use, and current regular exercise. RESULTS: Of 502 participants with at least one anxiety disorder, 84 (17 percent) reported no history of trauma, 233 (46 percent) had a history of trauma but no PTSD, and 185 (37 percent) met DSM-IV criteria for PTSD. Patients with PTSD reported a significantly greater number of current and lifetime medical conditions than did participants with other anxiety disorders but without PTSD. Primary care patients with PTSD were more likely to have had a number of specific medical problems, including anemia, arthritis, asthma, back pain, diabetes, eczema, kidney disease, lung disease, and ulcer. Possible explanations for the greater rates of medical conditions among participants with PTSD were examined as predictors in multiple regression. PTSD was found to be a stronger predictor of reported number of medical problems than trauma history, physical injury, lifestyle factors, or comorbid depression. CONCLUSIONS: These findings suggest that PTSD is associated with a higher rate of general medical complaints. PMID- 12096169 TI - Service use and costs among VA patients with schizophrenia taking risperidone or olanzapine. AB - OBJECTIVE: The authors compared the changes in health care utilization and costs between a group of patients with schizophrenia who started treatment with risperidone and a group that started treatment with olanzapine. METHODS: A retrospective analysis was conducted of patients with schizophrenia who were given an initial prescription for risperidone or for olanzapine between March 1997 and March 1999. The change in utilization and cost of inpatient hospitalizations, outpatient clinic visits, medications, and total health care services from one year before to one year after initiation of treatment for the two groups was compared. RESULTS: The risperidone and olanzapine groups comprised 325 and 285 patients, respectively. Total health care costs declined by $1,536 on average for patients in the risperidone group and increased by $4,217 on average for patients in the olanzapine group after initiation of treatment. The difference was statistically significant, and it was largely due to drug and hospitalization costs. Drug costs for patients in the risperidone group underwent a smaller increase than those for patients in the olanzapine group ($991 versus $1,861). Hospitalization costs decreased by $4,011 for patients in the risperidone group and increased by $1,423 for those in the olanzapine group. CONCLUSIONS: Total health care costs declined for patients taking risperidone and increased for patients taking olanzapine after treatment was initiated. PMID- 12096170 TI - Medical morbidity, mental illness, and substance use disorders. AB - OBJECTIVE: Previous research on the prevalence of medical disorders among adults with mental illness has been inconclusive. In general, studies have found higher rates among persons with mental illness, but these studies did not account for comorbid substance use disorders. The authors examined whether certain medical disorders are more prevalent among adults with severe mental illness and whether a comorbid substance use disorder increases prevalence beyond the effect of severe mental illness alone. METHODS: Administrative data from the Massachusetts Division of Medical Assistance were used in a cross-sectional observational study design. The sample consisted of 26,332 Medicaid beneficiaries 18 to 64 years of age. Of these, 11,185 had been treated for severe mental illness. Twelve-month prevalence rates were computed, and logistic regression was used to estimate the effect of a substance use disorder or another mental illness on the risk of having a medical disorder. RESULTS: Compared with Medicaid beneficiaries who were not treated for severe mental illness, those with severe mental illness had a significantly higher age- and gender-adjusted risk of the medical disorders considered in the study. Those with a comorbid substance use disorder had the highest risk for five of the disorders. CONCLUSIONS: The higher treated prevalence of certain medical disorders among adults with severe mental illness has three implications: substance use disorder is an important risk factor and requires early detection; integration of the treatment of medical disorders and severe mental illness should receive higher priority; and efforts should be made to develop specialized disease self-management techniques. PMID- 12096172 TI - Use of general medical services by VA patients with psychiatric disorders. AB - OBJECTIVE: Patients with psychiatric disorders, particularly those with severe mental illnesses, have high rates of undetected and untreated medical problems and substantially elevated mortality rates due to medical illness. The authors sought to develop a better understanding of the demographic, medical, and psychiatric characteristics of this population to inform efforts to improve the medical care of these persons. METHODS: Using Department of Veterans Affairs (VA) databases, the authors examined use of medical services by 175,653 patients who were treated in Southern California and Nevada during fiscal year 2000. Multivariate regression models were used to examine factors affecting receipt of any medical care and the number of medical visits. RESULTS: Patients with psychiatric diagnoses had fewer medical visits than other VA patients; the largest differences were seen for patients with severe mental illnesses. Patients who were younger and male had few visits. Patients with diabetes or hypertension who had been diagnosed as having schizophrenia, bipolar disorder, or an anxiety disorder had substantially fewer visits than those who did not have these psychiatric diagnoses. Patients older than 50 years were less likely to have any medical care if they had a diagnosis of a substance use, depressive, bipolar, or anxiety disorder or posttraumatic stress disorder. CONCLUSIONS: Use of medical care varies by psychiatric diagnosis. Young adults with schizophrenia and posttraumatic stress disorder and adults of all ages with bipolar disorder have an especially high risk of not receiving general medical services. Interventions to improve medical care may need to be tailored to the population being targeted. PMID- 12096171 TI - Co-occurrence of HIV and serious mental illness among Medicaid recipients. AB - OBJECTIVE: The authors estimated the treated period prevalence of HIV infection in the Medicaid population and the rate of HIV infection among persons with serious mental illness in that population. METHODS: This cross-sectional study used Medicaid claims data and welfare recipient files for persons aged 18 years or older for fiscal years 1994 through 1996 in Philadelphia. Claims data were merged with welfare recipient files to calculate the treated period prevalence of serious mental illness, defined as a schizophrenia spectrum disorder or a major affective disorder, and HIV infection in the Medicaid population and the odds of receiving a diagnosis of HIV infection among those who had a diagnosis of serious mental illness. RESULTS: The treated period prevalence of HIV infection was.6 percent among Medicaid recipients who did not have a diagnosis of a serious mental illness and 1.8 percent among those who did. After sex, age, race, and time on welfare during the study period were controlled for, patients with a schizophrenia spectrum disorder were 1.5 times as likely to have a diagnosis of HIV infection, and patients with a diagnosis of a major affective disorder were 3.8 times as likely. CONCLUSIONS: The rate of HIV infection is significantly elevated among persons with serious mental illness. Further studies are needed to determine modes of transmission of HIV, special treatment needs, and effective strategies for reducing the risk of HIV infection. PMID- 12096173 TI - Reliable self-report of health service use by individuals with serious mental illness. AB - The purpose of this study was to determine whether individuals with serious mental illness could consistently report their use of medical services. Twenty nine individuals with schizophrenia completed a test-retest protocol using a modified version of the Medical Expenditure Panel Survey; there was a one-week interval between the two administrations of the instrument. Consistency of reporting was generally strong and was strongest for items indicating whether a service had been used in the past six months, which suggests that people with schizophrenia can provide reliable information about their use of health services. Continued research in this area will increase understanding of the types and quality of medical care received by persons with schizophrenia. PMID- 12096174 TI - Health behaviors and health status of older women with schizophrenia. AB - The authors interviewed 43 women with schizophrenia who were 40 to 70 years of age about their health status, preventive health care, addictive behaviors, and comorbid medical conditions. Data were compared with those for age-matched samples from the general population. Thirty women in the study sample (71 percent) were overweight or obese, compared with 38 percent in the general population. Twenty-seven (63 percent) smoked cigarettes. Twenty-six women (62 percent) had received a mammogram in the past two years, compared with 86 percent in the general population. Rates of routine physical examinations and Pap tests did not differ markedly between the study sample and the general population. These results highlight the health impairments of older women who have schizophrenia. PMID- 12096176 TI - A comparison of guidelines for the treatment of schizophrenia. AB - Although the clinical and administrative rationales for the use of guidelines in the treatment of schizophrenia are convincing, meaningful implementation has been slow. Guideline characteristics themselves influence whether implementation occurs. The authors examine three widely distributed guidelines and one set of algorithms to compare characteristics that are likely to influence implementation, including their degree of scientific rigor, comprehensiveness, and clinical applicability (ease of use, timeliness, specificity, and ease of operationalizing). The three guidelines are the Expert Consensus Guideline Series' "Treatment of Schizophrenia"; the American Psychiatric Association's "Practice Guideline for the Treatment of Patients With Schizophrenia"; and the Schizophrenia Patient Outcomes Research Team (PORT) treatment recommendations. The algorithms are those of the Texas Medication Algorithm Project (TMAP). The authors outline the strengths of each and suggest how a future guideline might build on these strengths. PMID- 12096175 TI - Receipt of preventive medical services at psychiatric visits by patients with severe mental illness. AB - The authors used data from the National Ambulatory Medical Care Survey from 1992 to 1999 on 3,198 office visits to explore the extent to which psychiatrists provide clinical preventive medical services to patients with severe mental illness. Preventive services were provided during 11 percent of the visits. A multivariate analysis showed that preventive services were more likely to have been provided for patients with a chronic medical condition, for patients who were also seen by a nurse or other health provider during the visit, in rural areas, and during longer visits. Preventive services were less likely to have been provided during visits to health maintenance organizations and visits that took place later in the study period. PMID- 12096177 TI - One-year rehospitalization rates of patients discharged on atypical versus conventional antipsychotics. AB - This study examined one-year rehospitalization rates for patients who were discharged from Austin State Hospital between August 1, 1997, and July 31, 1998, while taking olanzapine, risperidone, or a conventional antipsychotic. Time to readmission was measured by the product-limit formula. Although conventional antipsychotics were associated with a lower rehospitalization rate, no significant difference in the one-year rehospitalization rate was observed between the groups. At 180 days after initial discharge, the patients who received olanzapine had a higher rate of rehospitalization than patients who were taking conventional antipsychotics. Conventional antipsychotics were associated with a lower one-year rehospitalization rate than risperidone or olanzapine. PMID- 12096178 TI - Involvement in 12-step programs among persons with dual diagnoses. AB - Although many people with substance use problems are referred to Alcoholics Anonymous (AA) or Narcotics Anonymous (NA), few studies have examined characteristics of persons who comply with such referrals. In particular, little is known about self-help meeting attendance by persons with dual diagnoses. This study examined rates of AA and NA attendance among 351 persons with dual diagnoses who were treated in a hospital setting. It also explored the relationship between diagnosis and meeting attendance. Ten months after hospitalization, the study participants demonstrated rates of AA or NA attendance that were similar to those of persons who were diagnosed as having substance use disorders without severe mental illness. However, patients with schizophrenia or schizoaffective disorders reported significantly fewer days of AA or NA meeting attendance. PMID- 12096179 TI - An integrative treatment for patients with persistent auditory hallucinations. PMID- 12096180 TI - Resident education in community psychiatry: a model of service-learning. PMID- 12096181 TI - Poverty, social problems, and serious mental illness. PMID- 12096182 TI - Poverty, social problems, and serious mental illness. PMID- 12096183 TI - Pastors' perceptions of mental disorders. PMID- 12096184 TI - High-bandwidth interactive telepsychiatry. PMID- 12096185 TI - Incorporating school mental health programs in SCHIP plans. State Children's Health Insurance Program. PMID- 12096187 TI - A deletion-generator compound element allows deletion saturation analysis for genomewide phenotypic annotation. AB - With the available eukaryotic genome sequences, there are predictions of thousands of previously uncharacterized genes without known function or available mutational variant. Thus, there is an urgent need for efficient genetic tools for genomewide phenotypic analysis. Here we describe such a tool: a deletion generator technology that exploits properties of a double transposable element to produce molecularly defined deletions at high density and with high efficiency. This double element, called P[wHy], is composed of a "deleter" element hobo, bracketed by two genetic markers and inserted into a "carrier" P element. We have used this P[wHy] element in Drosophila melanogaster to generate sets of nested deletions of sufficient coverage to discriminate among every transcription unit within 60 kb of the starting insertion site. Because these two types of mobile elements, carrier and deleter, can be found in other species, our strategy should be applicable to phenotypic analysis in a variety of model organisms. PMID- 12096188 TI - dELL is an essential RNA polymerase II elongation factor with a general role in development. AB - Several eukaryotic proteins increase RNA polymerase II (Pol II) transcription rates in vitro. The relative contributions of these factors to gene expression in vivo is unknown. The ELL family of proteins promote Pol II elongation in vitro, and the Drosophila ELL homolog (dELL) is associated with Pol II at sites of transcription in vivo. The purpose of this study was to test whether an ELL family protein is required for gene expression in vivo. We show that dELL is encoded by the Suppressor of Triplo-lethal locus [Su(Tpl)]. We have characterized seven distinct mutant alleles of Su(Tpl) and show that a dELL transgene rescues recessive lethality of Su(Tpl). Su(Tpl) mutations cause abnormal embryonic segmentation and dominantly modify expression of diverse genes during development. These data show that an ELL family elongation factor is essential, acts broadly in development, and is not functionally redundant to other elongation factors in vivo. PMID- 12096189 TI - A Tat subunit vaccine confers protective immunity against the immune-modulating activity of the human immunodeficiency virus type-1 Tat protein in mice. AB - The rational design of new therapies against HIV-1 necessitates an improved understanding of the mechanisms underlying the production of ineffective immune responses to HIV-1 in most infected individuals. This report shows that the CD8(+) T cell responses to gp120 were greatly diminished in mice vaccinated with a bicistronic gp120-Tat DNA vaccine, compared with those induced by a DNA vaccine encoding gp120 alone. The CD8(+) T cell responses induced by the latter included strong gp120-specific IFN-gamma secretion and protective antiviral immunity against challenge by a vaccinia-env pseudotype. The degree to which Tat influenced CD8(+) T cell responses depended on the bioactivity of Tat. Thus, a bicistronic DNA vaccine that expresses gp120 and a truncated Tat defective for LTR activation elicited strong IFN-gamma -secreting CD8(+) T cell responses to gp120 but conferred only marginal protection against the vaccinia-env challenge. The effect of Tat was completely blocked, however, by immunization with inactivated Tat protein before vaccination with the bicistronic gp120-Tat DNA vaccine. PMID- 12096190 TI - Early kinetic intermediate in the folding of acyl-CoA binding protein detected by fluorescence labeling and ultrarapid mixing. AB - Early conformational events during folding of acyl-CoA binding protein (ACBP), an 86-residue alpha-helical protein, were explored by using a continuous-flow mixing apparatus with a dead time of 70 micros to measure changes in intrinsic tryptophan fluorescence and tryptophan-dansyl fluorescence energy transfer. Although the folding of ACBP was initially described as a concerted two-state process, the tryptophan fluorescence measurements revealed a previously unresolved phase with a time constant tau = 80 micros, indicating formation of an intermediate with only slightly enhanced fluorescence of Trp-55 and Trp-58 relative to the unfolded state. To amplify this phase, a dansyl fluorophore was introduced at the C terminus by labeling an I86C mutant of ACBP with 5-IAEDANS [5 ((((2-iodoacetyl)amino)ethyl)amino)naphthalene-1-sulfonic acid]. Continuous-flow refolding of guanidine HCl-denatured ACBP showed a major increase in tryptophan dansyl fluorescence energy transfer, indicating formation of a partially collapsed ensemble of states on the 100-micros time scale. A subsequent decrease in dansyl fluorescence is attributed to intramolecular quenching of donor fluorescence on formation of the native state. The kinetic data are fully accounted for by three-state mechanisms with either on- or off-pathway intermediates. The intermediate accumulates to a maximum population of 40%, and its stability depends only weakly on denaturant concentration, which is consistent with a marginally stable ensemble of partially collapsed states with approximately 1/3 of the solvent-accessible surface buried. The findings indicate that ultrafast mixing methods combined with sensitive conformational probes can reveal transient accumulation of intermediate states in proteins with apparent two-state folding mechanisms. PMID- 12096191 TI - Molecular basis for the dichotomy in Plasmodium falciparum adhesion to CD36 and chondroitin sulfate A. AB - Plasmodium falciparum-infected erythrocytes adhere dichotomously to the host receptors CD36 and chondroitin sulfate A (CSA). This dichotomy is associated with parasite sequestration to microvasculature beds (CD36) or placenta (CSA), leading to site-specific pathogenesis. Both properties are mediated by members of the variant P. falciparum erythrocyte membrane protein 1 (PfEMP-1) family and reside on nonoverlapping domains of the molecule. To identify the molecular basis for the apparent dichotomy, we expressed various domains of PfEMP-1 individually or in combination and tested their binding properties. We found that the CD36 binding mode of the cysteine-rich interdomain region-1 (CIDR1) ablates the ability of the Duffy binding-like gamma domain to bind CSA. In contrast, neither a non-CD36-binding CIDR1 nor an intercellular adhesion molecule 1 binding domain had any affect on CSA binding. Our findings point out that interactions between different domains of PfEMP-1 can alter the adhesion phenotype of infected erythrocytes and provide a molecular basis for the apparent dichotomy in adhesion. We suggest that the basis for the dichotomy is structural and that mutually exclusive conformations of PfEMP-1 are involved in binding to CD36 or CSA. Furthermore, we propose a model explaining the requirement for structural dichotomy between placental and nonplacental isolates. PMID- 12096192 TI - The number of K(+) channels in the plasma membrane of guard cell protoplasts changes in parallel with the surface area. AB - The activity of the two dominant K(+) channels in the plasma membrane of Vicia faba guard cell protoplasts was examined during pressure-driven swelling. For this purpose, the K(+) currents and the membrane capacitance (C(m)) of guard cell protoplasts were recorded in parallel. A rise in C(m), reflecting an increase of the membrane surface area, was coupled to a proportional rise in conductance of both the K(+) inward and K(+) outward rectifier. The activation kinetics of the K(+) channels were not affected during this process. The quantitative and temporal coupling of C(m) and K(+) conductance can hence be interpreted as the result of the addition of active inward and outward rectifier K(+) channels to the plasma membrane during an increase in surface area. PMID- 12096193 TI - Short RNA duplexes produced by hydrolysis with Escherichia coli RNase III mediate effective RNA interference in mammalian cells. AB - Small interfering RNA (siRNA) has become a powerful tool for selectively silencing gene expression in cultured mammalian cells. Because different siRNAs of the same gene have variable silencing capacities, RNA interference with synthetic siRNA is inefficient and cost intensive, especially for functional genomic studies. Here we report the use of Escherichia coli RNase III to cleave double-stranded RNA (dsRNA) into endoribonuclease-prepared siRNA (esiRNA) that can target multiple sites within an mRNA. esiRNA recapitulates the potent and specific inhibition by long dsRNA in Drosophila S2 cells. In contrast to long dsRNA, esiRNA mediates effective RNA interference without apparent nonspecific effect in cultured mammalian cells. We found that sequence-specific interference by esiRNA and the nonspecific IFN response activated by long dsRNA are independent pathways in mammalian cells. esiRNA works by eliciting the destruction of its cognate mRNA. Because of its simplicity and potency, this approach is useful for analysis of mammalian gene functions. PMID- 12096196 TI - Is there more than one inflammatory phenotype in asthma? PMID- 12096195 TI - How prevalent is aspirin induced asthma? PMID- 12096194 TI - The outer membrane localization of the Neisseria gonorrhoeae MsrA/B is involved in survival against reactive oxygen species. AB - The PilB protein of Neisseria gonorrhoeae has been reported to be involved in the regulation of pilin gene transcription, but it also possesses significant homology to the peptide methionine sulfoxide reductase family of enzymes, specifically MsrA and MsrB from Escherichia coli. MsrA and MsrB in E. coli are able to reduce methionine sulfoxide residues in proteins to methionines. In addition, the gonococcal PilB protein encodes for both MsrA and MsrB activity associated with the repair of oxidative damage to proteins. In this work, we demonstrate that the PilB protein of Neisseria gonorrhoeae is not involved in pilus expression. Additionally, we show that wild-type N. gonorrhoeae produces two forms of this polypeptide, one of which contains a signal sequence and is secreted from the bacterial cytoplasm to the outer membrane; the other lacks a signal sequence and is cytoplasmic. Furthermore, we show that the secreted form of the PilB protein is involved in survival in the presence of oxidative damage. PMID- 12096197 TI - The prevalence of aspirin intolerant asthma (AIA) in Australian asthmatic patients. AB - BACKGROUND: Aspirin intolerant asthma (AIA) is a clinically distinct syndrome characterised by the precipitation of asthma attacks following the ingestion of aspirin and other non-steroidal anti-inflammatory drugs (NSAIDs). The prevalence of AIA among Australian asthmatic patients has not previously been reported. METHODS: Three populations were surveyed to establish the prevalence of AIA among Australian asthmatics. Two surveys were completed in patients recruited from the metropolitan area in Perth, Western Australia, one comprising 150 recruited from hospital based sources (hospital cohort) and the second comprising 366 from the membership of the Asthma Foundation of Western Australia (Asthma Foundation cohort). In a third study 1298 individuals were randomly selected from the rural community of Busselton in Western Australia. RESULTS: The prevalence of AIA in the hospital and Asthma Foundation cohorts was found to be 10.7% and 10.4%, respectively. Univariate analyses in the Asthma Foundation cohort indicated that AIA was associated with more severe asthma (OR = 2.4, 95% CI 1.18 to 4.86), nasal polyposis (OR=3.19, 95% CI 1.52 to 6.68), atopy (OR=2.96, 95% CI 1.48 to 5.89), sulfite sensitivity (OR=3.97, 95% CI 1.87 to 8.41), and sensitivity to wine (OR=3.27, 95% CI 1.65 to 6.47). Multivariate analyses indicated that atopy (OR=2.80, 95% CI 1.38 to 5.70), nasal polyposis (OR=3.39, 95% CI 1.57 to 7.29), and the number of asthma attacks in the previous 12 months (OR=1.20, 95% CI 1.02 to 1.42) were independent predictors for AIA, as was wine sensitivity (OR=2.20, 95% CI 1.02 to 4.72). The prevalence of AIA among asthmatic patients in the Busselton cohort was 10.9%. In addition, 2.5% of non-diagnosed asthmatics in this cohort reported asthma symptoms following aspirin ingestion. CONCLUSION: The prevalence of respiratory symptoms triggered by aspirin/NSAID use was found to be 10-11% in patients with asthma and 2.5% in non-asthmatics. Aspirin sensitivity appears to be a significant problem in the community and further investigations of the mechanisms of these responses and the possible link between this syndrome and other food and chemical sensitivities are required. PMID- 12096198 TI - Contrasting effects of allergen challenge on airway responsiveness to cysteinyl leukotriene D(4) and methacholine in mild asthma. AB - BACKGROUND: Cysteinyl leukotrienes (cysteinyl-LTs) have been implicated in the pathogenesis of allergen induced airway responses. Airway responsiveness (AR) to inhaled cysteinyl-LTs is dramatically increased following allergen challenge in animal studies. The effect in man has not been evaluated. METHODS: Ten mild steroid-naive asthmatic subjects with an isolated early asthmatic response (EAR) and 21 with an additional late asthmatic response (LAR) took part in a randomised controlled crossover study to assess AR to inhaled methacholine (MCh) and cysteinyl-LT D(4) (LTD(4)) 22 and 24 hours, respectively, after allergen challenge. Eight subjects had two further LTD(4) challenges separated by a 2 week washout period to assess the reproducibility of inhaled LTD(4) challenge. RESULTS: In subjects with an isolated EAR, non-significant mean (SE) increases in AR of 0.4 (0.4) doubling doses (DD) for MCh and 0.4 (0.5) DD for LTD(4) followed allergen challenge compared with control. A significant correlation between AR to MCh and LTD(4) followed both control (r=0.91, 95% CI 0.67 to 0.98; p=0.0002) and allergen challenge (r=0.79, 95% CI 0.32 to 0.95; p=0.0063). In subjects with an additional LAR there was a significant increase in AR to MCh (1.2 (0.3) DD, p=0.0005) following allergen challenge but no overall effect on AR to LTD(4) (0.69 (0.4) DD, p=0.11). A significant correlation between AR to MCh and LTD(4) was again observed (r=0.70; 95% CI 0.38 to 0.87; p=0.0004) following control, although it was reduced following allergen challenge (r=0.48; 95% CI 0.063 to 0.76; p=0.027). LTD(4) challenge was highly reproducible with a mean difference of 0.2 (0.3) DD between challenges. CONCLUSIONS: Allergen challenge significantly increases AR to inhaled MCh but not to LTD(4) in subjects with LAR. The lack of a comparable increase in AR to LTD(4) is surprising. Endogenous cysteinyl-LTs are produced in abundance following allergen challenge and may enhance AR to MCh or induce a degree of tachyphylaxis to LTD(4). PMID- 12096199 TI - Association of body mass index with the development of methacholine airway hyperresponsiveness in men: the Normative Aging Study. AB - BACKGROUND: The rising prevalence of asthma in developed nations may be associated with the rising prevalence of obesity in these same nations. The relationship between body mass index (BMI) and the development of an objective marker for asthma, methacholine airway hyperresponsiveness (AHR), was investigated in adult men. METHODS: Sixty one men who had no AHR at initial methacholine challenge testing but who developed AHR about 4 years later and 244 matched controls participated in the study. The effects of initial BMI and change in BMI on development of AHR were examined in conditional logistic regression models. RESULTS: Initial BMI was found to have a non-linear relationship with development of AHR. Compared with men with initial BMI in the middle quintile, men with BMI in the lowest quintile (BMI=19.8-24.3 kg/m(2)) and those with BMI in the highest quintile (BMI >29.4 kg/m(2)) were more likely to develop AHR: OR=7.0 (95% CI 1.8 to 27.7) and OR=10.0 (95% CI 2.6 to 37.9), respectively. These results remained significant after controlling for age, smoking, IgE level, and initial FEV(1). In addition, there was a positive linear relationship between change in BMI over the period of observation and the subsequent development of AHR. CONCLUSIONS: In this cohort of adult men, both a low BMI and a high BMI were associated with the development of AHR. For men with a low initial BMI the increased risk for development of AHR appears to be partly mediated by a gain in weight. The effect of BMI on AHR may suggest mechanisms in the observed associations between obesity and asthma. PMID- 12096200 TI - Nitric oxide in chronic airway inflammation in children: diagnostic use and pathophysiological significance. AB - BACKGROUND: The levels of exhaled and nasal nitric oxide (eNO and nNO) in groups of patients with inflammatory lung diseases are well documented but the diagnostic use of these measurements in an individual is unknown. METHODS: The levels of nNO and eNO were compared in 31 children with primary ciliary dyskinesia (PCD), 21 with non-CF bronchiectasis (Bx), 17 with cystic fibrosis (CF), 35 with asthma (A), and 53 healthy controls (C) using a chemiluminescence NO analyser. A diagnostic receiver-operator characteristic (ROC) curve for PCD using NO was constructed. RESULTS: The median (range) levels of nNO in parts per billion (ppb) in PCD, Bx, CF, and C were 60.3 (3.3-920), 533.6 (80-2053), 491.3 (31-1140), and 716 (398-1437), respectively; nNO levels were significantly lower in PCD than in all other groups (p<0.05). The median (range) levels of eNO in ppb in PCD, Bx, CF, A, and C were 2.0 (0.2-5.2), 5.4 (1.0-22.1), 2.6 (0.8-12.9), 10.7 (1.6-46.7), and 4.85 (2.5-18.3), respectively. The difference in eNO levels in PCD reached significance (p<0.05) when compared with those in Bx, A and C but not when compared with CF. Using the ROC curve, nNO of 250 ppb showed a sensitivity of 97% and a specificity of 90% for the diagnosis of PCD. CONCLUSIONS: eNO and nNO cannot be used diagnostically to distinguish between most respiratory diseases. However, nNO in particular is a quick and useful diagnostic marker which may be used to screen patients with a clinical suspicion of PCD. PMID- 12096201 TI - Increased levels of the chemokines GROalpha and MCP-1 in sputum samples from patients with COPD. AB - BACKGROUND: Patients with chronic obstructive pulmonary disease (COPD) have increased numbers of neutrophils and macrophages in their lungs. Growth related oncogene-alpha (GROalpha) attracts neutrophils, whereas monocyte chemoattractant protein-1 (MCP-1) attracts monocytes that can differentiate into macrophages. The aim of this study was to determine the concentration of GROalpha and MCP-1 in bronchoalveolar lavage (BAL) fluid and sputum from non-smokers, healthy smokers and patients with COPD, and to see if there was a correlation between the concentrations of these chemokines, lung function, and numbers of inflammatory cells. METHODS: BAL fluid and sputum from non-smokers (n=32), healthy smokers (n=36), and patients with COPD (n=40) were analysed for the presence of GROalpha and MCP-1 using ELISA. Cells counts were performed on the samples and correlations between the concentrations of these chemokines, lung function, and inflammatory cells observed. RESULTS: Median (SE) GROalpha and MCP-1 levels were significantly increased in sputum from patients with COPD compared with non smokers and healthy smokers (GROalpha: 31 (11) v 2 (2) v 3 (0.8) ng/ml; MCP-1: 0.8 (0.4) v 0.2 (0.1) v 0.1 (0.04) ng/ml, p<0.05), but not in BAL fluid. There were significant negative correlations between both GROalpha and MCP-1 levels in sputum and forced expiratory volume in 1 second (FEV(1)) % predicted (GROalpha: r=-0.5, p<0.001; MCP-1: r=-0.5, p<0.001), together with significant positive correlations between GROalpha and MCP-1 and neutrophil numbers in sputum (GROalpha: r=0.6, p<0.001; MCP-1: r=0.4, p<0.01). CONCLUSION: These results suggest that GROalpha and MCP-1 are involved in the migration of inflammatory cells, thus contributing to the inflammatory load associated with COPD. PMID- 12096202 TI - Relationship between nutritional status and lung function in cystic fibrosis: cross sectional and longitudinal analyses from the German CF quality assurance (CFQA) project. AB - BACKGROUND: The German cystic fibrosis (CF) quality assurance (CFQA) project is a patient registry for CF which was founded in 1995. Relevant clinical and laboratory data, respiratory function test results, complications, and CF treatments are entered into the database once a year for each patient. Using the database, a study was undertaken to elucidate the relationship between nutrition and lung function in a large patient cohort by cross sectional and longitudinal analysis. METHODS: A cohort of 3298 patients above 2 years of age was analysed. Patients were grouped according to the presence or absence of malnutrition (wasting and/or stunting). Cross sectional and longitudinal analyses over 2 and 3 years including mixed model analyses were performed. RESULTS: The prevalence of abnormal weight for height (<90% predicted) increased with age from 19% in children aged <6 years to 38% in adults with CF. Patients with malnutrition had significantly lower mean values of vital capacity, arterial oxygen tension (PO(2)), and forced expiratory volume in 1 second (FEV(1)) and higher serum IgG (p<0.05). Pseudomonas aeruginosa infection was also associated with decreased pulmonary function. Malnourished adolescents aged 12-18 years experienced a serious decline in FEV(1) of about 20% predicted, whereas mean FEV(1) values remained stable at above 80% predicted in adolescents of normal weight. Longitudinal follow up showed that malnourished patients of all ages and those with P aeruginosa infection had significantly worse lung function than their normally nourished counterparts and a greater yearly loss of FEV(1) % predicted. During 1 year of observation adolescents who experienced a >5% predicted decrease in weight for height had a concomitant mean loss of FEV(1) of 16.5% predicted during that year, whereas patients who gained relative weight had a parallel increase in FEV(1) of 2.1% predicted. CONCLUSIONS: These data emphasise the close relationship between nutrition, lung function, and clinical course in CF. Normal body weight and absence of P aeruginosa infection was associated with better preservation of lung function. PMID- 12096203 TI - Prevalence of sleep apnoea and snoring in hypertensive men: a population based study. AB - BACKGROUND: Several studies have reported an association between sleep disordered breathing (SDB) and hypertension (HT) but there is still a debate as to whether this is an effect of confounders. Some researchers have found an age dependent relationship between SDB and HT with higher risk at lower ages. A case-control study was performed in hypertensive men and non-hypertensive male controls matched for age and body mass index to assess whether there is an independent association between SDB and HT. If so, we further wanted to investigate whether this effect is age dependent. METHODS: An overnight sleep study was performed in a population based, age stratified sample of 102 hypertensive men aged 43-82 years and 102 non-hypertensive controls. RESULTS: Hypertensive subjects had a significantly higher prevalence of SDB than non-hypertensive subjects (apnoea hypopnoea index (AHI): 10.8 v 7.3; desaturation index (DI): 8.5 v 5.2; AHI >or=10: 37% v 24%, p<0.05; DI >or=10: 29% v 12%; lowest desaturation: mean (SD) 81.9 (7.3) v 84.7 (6.1), p<0.01). After adjusting for neck circumference and physical inactivity, DI >or=10 and lowest desaturation were still independent predictors of HT with adjusted odds ratios of 2.3 (95% CI 1.0 to 5.3) and 0.94 (95% CI 0.89 to 0.99), respectively. When the subjects were split into two groups according to age (<60 and >or=60 years), the influence of DI >or=10 on HT was strongest in the younger men (adjusted OR 4.3 (95% CI 1.0 to 19.3 v 2.1 (95% CI 0.7 to 6.5)) and the association between minimum oxygen saturation (SaO(2)) and HT reached statistical significance in the younger men only. CONCLUSION: SDB is more prevalent in men with HT than in controls. DI >or=10 and lowest desaturation are independent predictors of HT irrespective of confounders. The results indicate that the influence of SDB on HT is more pronounced in younger and middle aged men than in those above 60 years. PMID- 12096204 TI - Mid term effects of pulmonary thromboendarterectomy on clinical and cardiopulmonary function status. AB - BACKGROUND: Chronic thromboembolic pulmonary hypertension (CTEPH) can be successfully treated surgically by pulmonary thromboendarterectomy (PTE) but there are few data on mid-term cardiopulmonary function, particularly on exertion, and clinical benefits following pulmonary PTE. METHODS: A 2 year follow up study was undertaken of clinical status, haemodynamic and lung function indices, gas exchange, and exercise tolerance in 38 patients of mean (SD) age 50 (15) years who had undergone PTE. RESULTS: In-hospital mortality was about 10%. Before PTE all the patients were severely impaired (NYHA classes III-IV). There was no time difference in the improvement in the parameters: nearly all the improvement in cardiac output, gas exchange, and clinical status was achieved in the first 3 months as a result of the relief of pulmonary obstruction. At 3 months the percentage of patients with normal cardiac output and PaO(2) and of those with reduced clinical impairment increased to 97%, 59%, and 87%, respectively, without any further change. Only mean pulmonary artery pressure (mPAP), carbon monoxide transfer factor (TLCO), and exercise tolerance improved gradually during the second year, probably due to the recovery of the damaged small vessels. TLCO was overestimated before PTE but afterwards the trend was similar to that of mPAP. CONCLUSIONS: At mid term only a few patients did not have a satisfactory recovery because of lack of operative success, hypertension relapse, or the effect of preoperative hypertension on vessels in non-obstructed segments. Most of the patients, even the more compromised ones, had excellent long lasting results. PMID- 12096205 TI - Diaphragm plication following phrenic nerve injury: a comparison of paediatric and adult patients. AB - BACKGROUND: A study was undertaken to evaluate whether adults differ from children in the indications and outcome of diaphragmatic plication following phrenic nerve injury. METHODS: A retrospective study was performed of 21 patients, 10 below the age of 5 and 11 older than 37 years. The indication for surgery for all the children was failure to wean from ventilatory support. The indications for surgery in the adult group were ventilator dependency (n=4) and symptomatic dyspnoea (n=7). All patients had at least one imaging study confirming diaphragmatic paralysis. The American Thoracic Society (ATS) dyspnoea scale, pulmonary function tests, and quantitative pulmonary perfusion scans were used as evaluation parameters. At surgery the diaphragm was centrally plicated. RESULTS: One child died immediately after surgery due to irreversible heart failure and two children died within 2 months of surgery from ongoing complications of their original condition. These three patients were considered as selection failures. Seven children were weaned from ventilatory support within a median of 4 days (range 2-140). Only one of four ventilated adults was successfully weaned. Seven adults who underwent surgery for chronic symptoms had a marked subjective improvement of 2-3 levels in the ATS dyspnoea scale. Pulmonary function studies in the seven symptomatic adults showed a 40% improvement above baseline. Severely asymmetrical perfusion scans reverted to a normal pattern after plication. CONCLUSIONS: Diaphragmatic plication offers a significant benefit to children with diaphragmatic paralysis and should be performed early to facilitate weaning from mechanical ventilation. While plication is of limited benefit in weaning ventilated adults, it results in significant subjective and objective lifetime improvement in non-ventilated symptomatic adults. PMID- 12096206 TI - Molecular epidemiology of tuberculosis in London 1995-7 showing low rate of active transmission. AB - BACKGROUND: Tuberculosis notification rates for London have risen dramatically in recent years. Molecular typing of Mycobacterium tuberculosis has contributed to our understanding of the epidemiology of tuberculosis throughout the world. This study aimed to assess the degree of recent transmission of M tuberculosis in London and subpopulations of the community with high rates of recent transmission. METHODS: M tuberculosis isolates from all persons from Greater London diagnosed with culture positive tuberculosis between 1 July 1995 and 31 December 1997 were genetically fingerprinted using IS6110 restriction fragment length polymorphism (RFLP) typing. A structured proforma was used during record review of cases of culture confirmed tuberculosis. Cluster analysis was performed and risk factors for clustering were examined in a univariate analysis followed by a logistic regression analysis with membership of a cluster as the outcome variable. RESULTS: RFLP patterns were obtained for 2042 isolates with more than four copies of IS6110; 463 (22.7%) belonged to 169 molecular clusters, which ranged in size from two (65% of clusters) to 12 persons. The estimated rate of recent transmission was 14.4%. Young age (0-19 years) (odds ratio (OR) 2.65, 95% confidence interval (CI) 1.59 to 4.44), birth in the UK (OR 1.55, 95% CI 1.04 to 2.03), black Caribbean ethnic group (OR 2.19, 95% CI 1.15 to 4.16), alcohol dependence (OR 2.33, 95% CI 1.46 to 3.72), and streptomycin resistance (OR 1.82, 95% CI 1.15 to 2.88) were independently associated with an increased risk of clustering. CONCLUSIONS: Tuberculosis in London is largely caused by reactivation or importation of infection by recent immigrants. Newly acquired infection is also common among people with recognised risk factors. Preventative interventions and early diagnosis of immigrants from areas with a high incidence of tuberculosis, together with thorough contact tracing and monitoring of treatment outcome among all cases of tuberculosis (especially in groups at higher risk of recent infection), remains most important. PMID- 12096207 TI - Raised plasma concentrations of alpha-defensins in patients with idiopathic pulmonary fibrosis. AB - BACKGROUND: Neutrophils are thought to play an important role in the pathogenesis of idiopathic pulmonary fibrosis (IPF). Human neutrophils contain antimicrobial and cytotoxic peptides in the azurophil granules which belong to a family of mammalian neutrophil peptides named alpha-defensins. A study was undertaken to investigate the role of alpha-defensins in the pathogenesis of IPF. METHODS: The concentrations of alpha-defensins (human neutrophil peptides (HNPs) 1, 2, and 3) in plasma and bronchoalveolar lavage (BAL) fluid of 30 patients with IPF and 15 healthy subjects were measured by radioimmunoassay. RESULTS: The concentrations of alpha-defensins in plasma, but not in BAL fluid, were significantly higher in IPF patients than in controls. BAL fluid concentrations of interleukin (IL)-8 in patients with IPF, which were significantly higher than in controls, correlated with those of alpha-defensins. An inverse relationship was seen between plasma alpha-defensin levels and the arterial oxygen tension (PaO(2)) and pulmonary function (vital capacity (%VC), forced expiratory volume in 1 second (FEV(1)), and carbon monoxide transfer factor (%TLCO)) in patients with IPF. Plasma levels of alpha-defensins also correlated with the clinical course in IPF patients with an acute exacerbation. Immunohistochemically, positive staining was observed inside and outside neutrophils in the alveolar septa, especially in dense fibrotic areas. CONCLUSION: These findings suggest that alpha-defensins play an important role in the pathogenesis of IPF, and that the plasma alpha-defensin level may be a useful marker of disease severity and activity. PMID- 12096208 TI - P-selectin upregulation in bleomycin induced lung injury in rats: effect of N acetyl-L-cysteine. AB - BACKGROUND: A number of adhesion molecules are involved in the process of neutrophil infiltration into the lung. P-selectin is one of these neutrophil endothelial cell adhesion molecules. A study was undertaken to examine the involvement of P-selectin in the development of bleomycin induced inflammation and the ability of N-acetyl-L-cysteine to reduce the potential expression of this selectin in rats. METHODS: N-acetyl-L-cysteine (3 mmol/kg po) was administered daily for seven days prior to bleomycin administration (2.5 U/kg). The kinetics of P-selectin expression and the effect of N-acetyl-L-cysteine after bleomycin treatment were measured using radiolabelled antibodies. P-selectin localisation was evaluated by immunohistochemistry and neutrophil infiltration was assessed by myeloperoxidase activity. RESULTS: Bleomycin administration resulted in an upregulation of P-selectin at 1 hour, returning to baseline at 3 hours. Myeloperoxidase activity showed a significant increase at 6 hours after bleomycin administration that lasted for 3 days. N-acetyl-L-cysteine treatment completely prevented these increases. CONCLUSION: Upregulation of P-selectin in the lung is associated with neutrophil recruitment in response to bleomycin. The beneficial effect of N-acetyl-L-cysteine on bleomycin induced lung injury may be explained in part by the prevention of neutrophil recruitment in the inflammatory stage of the disease. PMID- 12096212 TI - Pulmonary sarcoidosis and the acute respiratory distress syndrome (ARDS). AB - A 50 year old man presented with 3 weeks of exertional dyspnoea. His chest radiograph on admission revealed diffuse bilateral interstitial infiltrates. He did not respond to antibiotics but subsequently improved on high dose corticosteroids. Bronchoscopic examination with transbronchial biopsy specimens revealed the presence of non-necrotising granulomas. This case demonstrates an unusual clinical presentation of life threatening pulmonary sarcoidosis characterised by the development of acute respiratory distress syndrome (ARDS) with acute respiratory failure. PMID- 12096209 TI - The pulmonary physician in critical care * 7: ventilator induced lung injury. PMID- 12096213 TI - Transudates and exudates. PMID- 12096214 TI - BTS guidelines on CAP. Community acquired pneumonia. PMID- 12096210 TI - Non-eosinophilic asthma: importance and possible mechanisms. AB - There is increasing evidence that inflammatory mechanisms other than eosinophilic inflammation may be involved in producing the final common pathway of enhanced bronchial reactivity and reversible airflow obstruction that characterises asthma. A review of the literature has shown that, at most, only 50% of asthma cases are attributable to eosinophilic airway inflammation. It is hypothesised that a major proportion of asthma is based on neutrophilic airway inflammation, possibly triggered by environmental exposure to bacterial endotoxin, particulate air pollution, and ozone, as well as viral infections. If there are indeed two (or more) subtypes of asthma, and if non-eosinophilic (neutrophil mediated) asthma is relatively common, this would have major consequences for the treatment and prevention of asthma since most treatment and prevention strategies are now almost entirely focused on allergic/eosinophilic asthma and allergen avoidance measures, respectively. It is therefore important to study the aetiology of asthma further, including the underlying inflammatory profiles. PMID- 12096211 TI - Evolving concepts on the value of adenosine hyperresponsiveness in asthma and chronic obstructive pulmonary disease. AB - Adenosine is a purine nucleoside which mediates a variety of cellular responses relevant to asthma and COPD through interaction with specific receptors. Administration of adenosine by inhalation to patients with asthma and COPD is known to cause concentration related bronchoconstriction. Responses elicited by this purine derivative in asthma and COPD should not be considered as a mere reflection of non-specific airways hyperresponsiveness. Evaluation of airways responsiveness by adenosine induced bronchoconstriction may be valuable in differentiating asthma from COPD, monitoring of anti-inflammatory treatment in asthma, surveying disease progression, and assessing disease activity in relation to allergic airways inflammation. PMID- 12096215 TI - Mycobacterium xenopi. PMID- 12096216 TI - Cytoskeletal proteins and gene regulation: form, function, and signal transduction in the nucleus. AB - The cytoskeleton is important for controlling cell shape and cell motility and for organizing signaling complexes. Additionally, connections are being found between cytoskeletal proteins and the regulation of gene expression in the nucleus. De Lanerolle and Cole discuss new insights from plants that show that a plant member of the COOH-terminal binding protein (CtBP) and the brefeldin A adenosine diphosphate (ADP)-ribosylated subtrates (BARS) proteins, ANGUSTIFOLIA, may be a regulator of both the microtubule-based cytoskeleton and a transcriptional regulator. Additionally, they describe how actin and myosins may play similar roles in regulating gene expression in the nucleus of mammalian cells. PMID- 12096218 TI - The muscle in fibromyalgia. PMID- 12096217 TI - Microtubule-actin cross-talk at focal adhesions. AB - Focal adhesions are dynamic structures in which traction forces are exerted against the substratum during cell migration and are sites for the organization of signaling complexes. Palazzo and Gundersen discuss how focal adhesions may also be the site of cross-talk between the actin-based and microtubule-based cytoskeletons. Microtubules appear to deliver factors that can regulate the formation and dissolution of focal adhesions, whereas focal adhesions contribute to microtubule localization and stability. PMID- 12096219 TI - Genetic studies of disorders of calcium crystal deposition. PMID- 12096220 TI - Superior mesenteric artery blood flow in systemic sclerosis patients. AB - OBJECTIVE: Intestinal involvement is frequently observed in systemic sclerosis (SSc) and is associated with malnutrition and a decreased survival rate. Vascular lesions are claimed to underlie and precede these changes. The aim of this study was to establish whether a reduced mesenteric blood flow was present in SSc patients with no signs or symptoms of small bowel involvement. METHODS: Superior mesenteric artery (SMA) blood flow in the fasting state was measured by colour Doppler ultrasonography in 27 SSc patients and in 25 controls. The effect of a balanced liquid meal on mesenteric blood flow was measured in six matched patients and controls. RESULTS: In fasting SSc patients, there were reductions in mean SMA diameter (P<0.001), blood flow (213+/-92 vs 398+/-125 ml/min in controls, P<0.0001) and pulsatility index (3.49+/-1.0 vs 4.13+/-0.97 in controls, P<0.07). In both groups, the meal increased basal flow values and the differences between controls and patients in the fasting state were not significant. CONCLUSIONS: In the absence of symptoms of small bowel involvement, reversible SMA vasoconstriction is demonstrable in the fasting state in SSc patients. PMID- 12096221 TI - Methotrexate in the treatment of rheumatoid arthritis. I. In vitro effects on cells of the osteoblast lineage. AB - OBJECTIVE: Low-dose methotrexate (MTX) is often used in the treatment of rheumatoid arthritis (RA). To be effective, treatment must be long-term, and there are concerns that MTX may impair bone formation in a population already predisposed to osteoporosis. The purpose of this investigation was to determine the direct effects of MTX at clinically relevant doses on the growth and differentiation of human cells of the osteoblast (bone-forming) lineage. METHODS: Cells derived from the marrow stroma (BMSC) and trabecular surfaces [human bone derived cells (HBDC)] of adult ribs were cultured in the absence or presence of MTX (1-1000 nM). To promote the differentiation and further maturation of cells of the osteoblast lineage, one half of the cultures were treated additionally with 10 nM dexamethasone (Dx). RESULTS: In cultures of BMSC, treatment with MTX (+/-Dx) did not affect the total number of colonies that formed or the expression of the developmental markers STRO-1 and alkaline phosphatase (AP). At concentrations > or =10 nM, however, there was a statistically significant reduction in the number of cells harvested at the end of primary culture. In cultures of HBDC, treatment with MTX (in the presence of Dx) did not affect cell number or the expression of AP. CONCLUSIONS: At concentrations > or =10 nM, treatment with MTX inhibits the proliferation of primitive marrow stromal cells, but not their ability to undergo osteogenic differentiation. The proliferation and further maturation of cells of the osteoblast lineage is not affected by treatment with MTX. These findings are reassuring for clinicians using MTX in the treatment of RA. PMID- 12096223 TI - Illiteracy in rheumatoid arthritis patients as determined by the Rapid Estimate of Adult Literacy in Medicine (REALM) score. AB - OBJECTIVES: To determine the prevalence of illiteracy in a cohort of rheumatoid arthritis (RA) patients and the impact of illiteracy on disease severity and function. METHODS: We performed a prospective cross-sectional study with case record review of 127 consecutive patients with RA attending one centre. All patients completed the Rapid Estimate of Adult Literacy in Medicine (REALM) screening test. This 66-word recognition test provides an estimate of reading level in less than 3 min. Demographic data were collected by interview and case record review. Function was assessed with the Health Assessment Questionnaire (HAQ) and depression with the Hospital Anxiety and Depression (HAD) scale, both sent prior to clinic attendance. Social deprivation was assessed with the Carstairs index. RESULTS: Four patients refused to participate. Of these, three stated they were unable to read. Ninety-seven women and 26 men agreed to be interviewed. All but two were Caucasian. Median age was 56 yr (range 19-77 yr) and median disease duration was 10 yr (range 1-60 yr). Median number of previous disease-modifying anti-rheumatic drugs (DMARDs) was two. Eighteen (15%) patients were functionally illiterate, with a REALM score of less than 60. Sex, age, disease duration and numbers of joint replacements and previous DMARDs were not influenced by illiteracy. Illiteracy led to more anxiety (P=0.011), but did not affect HAQ score (P>0.5). Illiteracy was more common in the deprived (P=0.0064). Illiterate patients had three times more hospital visits compared with age- and sex-matched RA controls over the previous 12 months. CONCLUSIONS: One in six of our patient population are illiterate and would struggle to cope with patient education materials and prescription labels. These patients had significantly more hospital visits but equal function, suggesting that additional resources be directed towards these individuals. The REALM test is quick and easy to administer and allows us to identify patients who may require more appropriate literature. PMID- 12096222 TI - Methotrexate in the treatment of rheumatoid arthritis. II. In vivo effects on bone mineral density. AB - OBJECTIVE: To determine the effect of methotrexate (MTX) on bone mineral density (BMD) in rheumatoid arthritis (RA). METHODS: One hundred and sixteen non-steroid treated RA subjects (90 women) were studied in a prospective, longitudinal, non randomized study. Subjects started MTX (n=36) or sulphasalazine (n=23) or continued long-term (>5 yr) treatment with MTX (n=28) or other disease-modifying anti-rheumatic drugs (n=29). BMD was estimated at entry and after 1 yr. Markers of bone turnover were measured at entry and at 1 yr, and additionally at 3 and 6 months in those starting treatment. Bone biopsies were taken before and after MTX treatment in four subjects. The primary outcome was change in BMD Z score and secondary outcomes were changes in bone turnover markers and bone formation by histomorphometry. RESULTS: Univariate analysis of covariance found that MTX at baseline was associated with reduced BMD at the femoral neck. However, femoral neck BMD was also associated with radiological damage score for the hand. Multivariate analysis and discriminant analysis of the subset of post-menopausal women showed that reduced bone density associated with MTX was due to confounders such as disease activity. There was no adverse effect of MTX on bone turnover markers or on measures of bone formation in biopsies. CONCLUSIONS: No adverse effect of low-dose MTX (mean 10 mg/week) on bone formation in RA was found. PMID- 12096224 TI - Measured ambulation and self-reported health status following total joint replacement for the osteoarthritic knee. AB - OBJECTIVE: To quantify the increase in ambulation produced by total knee replacement for osteoarthritis (OA) of the knee and to compare this with questionnaire-derived data. METHODS: We compared the measured ambulatory activity and self-reported Nottingham Health Profile (NHP) data of 19 subjects with OA of the knee before and after total knee replacement (TKR). RESULTS: Subjects were considerably restricted in their measured activity before operation compared with subjects with less severe disease. At 6 months the activity had increased by an average of 79% (P=0.02). The pain scale of the NHP had significantly improved at 3 months, as had the mobility scale. Between 3 and 6 months, however, at the same time the measured activity was increasing, self-reported mobility declined. CONCLUSIONS: Replacement of a knee for OA is an effective way of improving ambulation. At 6 months the average increase amounted to 79%. Self-reported mobility did not correlate with mobility measured objectively. PMID- 12096226 TI - Total costs and predictors of costs in individuals with early inflammatory polyarthritis: a community-based prospective study. AB - OBJECTIVE: To estimate the health service, non-health service and total costs and predictors of costs in individuals with early inflammatory polyarthritis (IP). METHODS: We conducted a prospective longitudinal study over a 6-month period. The participants were a random sample of 133 individuals who had enrolled with the community-based Norfolk Arthritis Register (NOAR) database between 1994 and 1999. The main outcome measures were the mean (per person) 6-month health service cost, non-health-service cost and total cost associated with IP. RESULTS: One hundred and fifteen of the 133 individuals who were recruited into the study completed 6 months of follow-up. The mean 6-month total cost was estimated to be 2800 pounds sterling per person, of which 14% was health service costs and the remainder non health-service costs. Higher total costs were associated with lower health status and rheumatoid factor positivity. CONCLUSIONS: Early IP has a considerable impact on both the health-care system and, more importantly, society. Non-health-service costs (i.e. costs incurred by the individual with the disease, their family and friends) account for a substantial proportion (86%) of the total costs associated with early IP. PMID- 12096225 TI - A 588-gene microarray analysis of the peripheral blood mononuclear cells of spondyloarthropathy patients. AB - OBJECTIVES: To identify genes which are more highly expressed in the peripheral blood mononuclear cells (PBMC) of patients with spondyloarthropathy (SpA), rheumatoid arthritis (RA) and psoriatic arthritis (PsA), in comparison to normal subjects. METHODS: A 588-gene microarray was used as a screening tool to select a panel of such genes from PBMC of these subjects and of normal subjects. Results were then validated by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The following genes were more highly expressed in arthritis patients than in normal subjects: macrophage differentiation marker MNDA (myeloid nuclear differentiation antigen), MRP8 and MRP14 (migratory inhibitory factor-related proteins); signalling molecules JAK3 (janus kinase 3) and MAP kinase p38 (mitogen activated protein kinase); receptors TNFR2/p75, C-C-chemokine receptor type 1 (CCR1), C-X-C-chemokine receptor type 4 (CXCR4) and integrin beta1; and the cytokines/chemokines interleukin (IL) 1beta and IL-8. Expression of CXCR4 was unexpectedly high among all arthritis subjects. Using RT-PCR, ELISA and immunohistology, expression of stromal cell-derived factor 1 (SDF-1) was demonstrated in arthritis joints. CONCLUSIONS: The CXCR4/SDF-1 is a potential pro inflammatory axis for RA, PsA and SpA. PMID- 12096227 TI - Dynamics of mycobacterial HSP65-induced T-cell cytokine expression during oral tolerance induction in adjuvant arthritis. AB - OBJECTIVE: To investigate whether oral administration of mycobacterial heat-shock protein 65 (HSP65) during adjuvant arthritis (AA) induces regulatory cells and cytokines. METHODS: AA was induced in Lewis rats and from the time of disease onset HSP65 in the presence of soya bean trypsin inhibitor (STI) was administered orally every other day. The number of splenic CD4+CD25+ T cells and antigen induced cytokine mRNA expression were determined. RESULTS: Oral treatment with HSP65/STI reduced AA symptoms. After one feeding of HSP65/STI, the number of CD4+CD25+ splenic T cells increased and HSP65-specific T cells expressed increased levels of interferon gamma and interleukin 10. After two feedings, the expression of interleukin-10 mRNA remained increased, whereas there was low expression of interferon gamma mRNA. The number of CD4+CD25+ splenic T cells remained increased. CONCLUSIONS: Oral treatment with HSP65/STI after AA onset reduces disease symptoms via dynamic changes in the number of CD4+CD25+ splenocytes and in antigen-induced cytokine production. PMID- 12096228 TI - Risk of myelotoxicity with intravenous cyclophosphamide in patients with systemic lupus erythematosus. AB - OBJECTIVES: To determine the incidence of serious myelotoxicity from intravenous cyclophosphamide (IC) in systemic lupus erythematosus (SLE). METHODS: In a retrospective study, white blood cell (WBC) counts with differential and platelet counts were determined in 92 SLE patients (96 courses) given 1623 doses of IC. RESULTS: Only one patient developed a total leucocyte count <1000/mm3, one developed a neutrophil count <500/mm3, two had a lymphocyte count <100/mm3 and no patients had platelet counts <50000/mm3 during follow-up. The risk of a neutrophil count <500/mm3 was 0.06 per 100 visits [95% confidence interval (CI) 0.00-0.34]. Two patients discontinued IC due to neutropenia [rate of 0.12 per 100 doses (95% CI 0.01-0.44)]. No clinical consequences were recorded in conjunction with low blood cell counts. In multivariate models, both the cumulative number of IC doses and European Consensus Lupus Activity Measurement (ECLAM) score affected neutrophil and lymphocyte counts adversely. For neutrophils, lowering the ECLAM score by 1 point counteracted four additional doses of IC after adjusting for steroid dose. CONCLUSIONS: IC and SLE disease activity have independent effects in lowering white blood cell counts, but serious myelotoxicity of IC is uncommon. PMID- 12096229 TI - Role of cholecystokinin-8 in nerve growth factor and nerve growth factor mRNA expression in carrageenan-induced joint inflammation in adult rats. AB - OBJECTIVES: The aim of our study was to investigate the role of cholecystokinin-8 (CCK-8), which is able to induce the synthesis of nerve growth factor (NGF), in the joint inflammation of carrageenan-injected rats. METHODS: Adult rats were injected in the ankle joint with carrageenan, with or without CCK-8 or a CCK receptor antagonist (proglumide), and tissue swelling, NGF levels and NGF mRNA expression were assessed. RESULTS: Expression of NGF and NGF mRNA increased transiently after carrageenan injection. This effect was not altered by CCK-8 injection but was inhibited by the CCK receptor antagonist. The decrease in NGF level after treatment with the antagonist was concurrent with an increase in paw swelling. CONCLUSIONS: The results demonstrate that, whereas CCK-8 has no anti inflammatory action in carrageenan-injected animals, proglumide induces a worsening of inflammation and reduces the expression of both NGF and NGF mRNA in inflamed ankle joints. Our data point to a regulatory action of CCK-8 on NGF synthesis during acute synovitis and suggest a role for NGF in the healing phase of inflammation. PMID- 12096230 TI - The prevalence of rheumatoid arthritis in the United Kingdom: new estimates for a new century. AB - BACKGROUND: It is 40 yr since the last age- and sex-specific estimates of the prevalence of rheumatoid arthritis (RA) for the UK were published. Since then the classification criteria for RA have been revised and there has been evidence of a fall in the incidence of RA, especially in women. OBJECTIVES: To estimate the age and sex-specific point prevalence of RA (defined as fulfilment of a modification of the 1987 ACR classification criteria for RA on the day of assessment). The estimate was made in the primary care setting in Norfolk, UK. METHODS: A stratified random sample was drawn from seven age and gender bands. The 7050 individuals selected were mailed a screening questionnaire. Positive responders were invited to attend for a clinical examination. The sample was matched against the names in the Norfolk Arthritis Register (NOAR), a register of incident cases of inflammatory polyarthritis which has been in existence since 1990. RESULTS: The overall response rate was 82%. Sixty-six cases of RA were identified. Extrapolated to the population of the UK, the overall minimum prevalence of RA is 1.16% in women and 0.44% in men. A number of incident cases of RA previously notified to NOAR were not identified as cases in the survey because they had entered into treatment-induced remission. In addition, some cases who failed to attend for examination had significant disability. These prevalence figures are therefore an underestimate. CONCLUSIONS: The prevalence of RA in women, but not in men, in the UK may have fallen since the 1950s. PMID- 12096231 TI - Selective COX-2 inhibition prevents proinflammatory cytokine-induced cartilage damage. AB - OBJECTIVES: This study evaluated the in vitro effect of the selective cyclooxygenase-2 (COX) inhibitor celecoxib on cartilage matrix turnover under normal and inflammatory conditions. METHODS: Healthy human articular cartilage tissue alone, in co-culture with peripheral blood mononuclear cells (PBMC) or in the presence of interleukin 1 (IL-1beta) plus tumour necrosis factor alpha (TNF alpha) was cultured for 7 days in the presence of celecoxib. Changes in cartilage matrix turnover were measured. RESULTS: No direct effects of celecoxib on healthy normal cartilage were found. Both PBMC and IL-1beta plus TNF-alpha induced strong inhibition of cartilage proteoglycan synthesis and significant enhancement of the release of proteoglycans, diminishing proteoglycan content. Celecoxib was able to reverse these adverse effects up to complete normalization. CONCLUSIONS: The results suggest that, under the influence of inflammation, COX-2 is up-regulated, which results in disturbance of cartilage matrix turnover. Celecoxib, by diminishing COX-2 activity, prevents these adverse effects without having a direct effect on healthy cartilage. PMID- 12096232 TI - Diagnostic tests for rheumatoid arthritis: comparison of anti-cyclic citrullinated peptide antibodies, anti-keratin antibodies and IgM rheumatoid factors. AB - OBJECTIVES: To examine the value of anti-cyclic citrullinated peptide (anti-CCP) antibodies, anti-keratin antibodies (AKA) and immunoglobulin M rheumatoid factors (IgM RF) in discriminating between rheumatoid arthritis (RA) and other rheumatic diseases, and to determine whether the clinical manifestations or severity of erosions in RA are associated with anti-CCP positivity. METHODS: In a cross sectional study, we determined the concentrations or titres of these three markers in 179 RA patients and 50 controls. Erosions were quantified using the Larsen score in 129 patients. RESULTS: Sensitivity was highest for IgM RF (75%), followed by anti-CCP antibodies (68%) and AKA (46%). Specificity was highest for anti-CCP antibodies (96%), followed by AKA (94%) and IgM RF (74%). A correlation with clinical manifestations and severity of erosions was observed mainly for IgM RF positivity. CONCLUSIONS: With their excellent specificity, anti-CCP antibodies can be useful in establishing the diagnosis of RA, but IgM RF is a better predictor of disease severity. PMID- 12096233 TI - Rapid diagnosis of inflammatory synovial fluid with reagent strips. AB - OBJECTIVE: To determine the usefulness of reagent test strips for screening inflammatory synovial fluid (SF). METHODS: Consecutive patients undergoing diagnostic arthrocentesis, attending the Department of Rheumatology of a large tertiary care hospital were evaluated. All SF specimens obtained were tested using two techniques: (i) white blood cell (WBC) count with the differential according to standard practice (which is considered the gold standard) (an inflammatory SF was defined as a WBC count > or =2000 cells/mm3); and (ii) reagent strips used to test urine (Multistix 8 SG, Bayer Diagnostics) for the presence of leucocytes (a positive test was defined as a strip showing more than a trace for leucocytes). Sensitivity, specificity, predictive values and likelihood ratio (LR) of the reagent strip in diagnosing inflammatory SF were determined. RESULTS: Two hundred and eight samples of SF were tested. The results of using the reagent strip were: sensitivity 76.9% (95% CI, 66.0-85.7%), specificity 86.9% (95% CI, 79.9-92.2%); positive LR, 5.88 (95% CI, 3.71-9.31) and negative LR, 0.27 (95% CI, 0.18-0.40). In 13 of the 19 false negative results, the differential cell count showed a predominance (> or =50%) of lymphocytes. CONCLUSION: This study suggests that, in daily practice, the evaluation of SF by reagent strips could be of use to discriminate between inflammatory and non inflammatory SF. PMID- 12096234 TI - The synergistic value of focus score and IgA% score of sublabial salivary gland biopsy for the accuracy of the diagnosis of Sjogren's syndrome: a 10-year comparison. AB - OBJECTIVE: Increasing the accuracy of the diagnosis of Sjogren's syndrome (SS) by placing emphasis on objective findings such as the presence of anti-Ro and anti La autoantibodies and abnormal salivary gland tissue (SGT) histology is a current issue. In order to obtain optimal disease sensitivity and specificity of SGT findings, histological and immunohistological SGT examinations were compared. The first describes the extent of the lymphocytic infiltrate as a focus score (LFS), whereas the latter describes the composition of the infiltrate as a percentage of IgA-containing plasma cells (IgA%). METHODS: Both the LFS and IgA% score were assessed in 279 SGT biopsies taken from patients with symptoms suggestive of SS. In case histological conclusions did not match immunohistological conclusions patients were assigned to so-called mismatch groups. Patients in the mismatch groups were further classified using objective, serological parameters [rheumatoid factor (RF), anti-Ro, anti-La, anti-nuclear antibodies, gammaglobulin level]. RESULTS: In 249 samples (89%), LFS and IgA% resulted in the same conclusion. Within this group a total of 63 SGT samples (25%) were characteristic for SS showing LFS >1.0 and IgA% <70. In the mismatch groups after serological classification, both false positive as well as false negative scores were observed less frequently for IgA% as compared with LFS (50 vs 75% and 25 vs 50%, respectively). CONCLUSIONS: Additional immunohistological SGT examination provides greater disease sensitivity and specificity than histological SGT examination alone, thereby increasing accuracy of SS diagnosis. PMID- 12096236 TI - Progressive multifocal leucoencephalopathy isolated to the posterior fossa in a patient with systemic lupus erythematosus. PMID- 12096235 TI - Professor Sir John Charnley (1911-1982). PMID- 12096238 TI - Necrotizing fasciitis in a patient with rheumatoid arthritis. PMID- 12096237 TI - A rare case of recurrent post-streptococcal myalgia. PMID- 12096239 TI - Prolonged arthritis associated with Sindbis-related (Pogosta) virus infection. PMID- 12096240 TI - Association of accelerated erosive rheumatoid arthritis with a polymorphism that alters NF-kappaB binding to the TNF promoter region. PMID- 12096241 TI - A case of Behcet's disease with pulmonary arterial aneurysm and secondary amyloidosis. PMID- 12096242 TI - Extensor tendon rupture. PMID- 12096243 TI - Re: The association of the involvement of financial compensation with the outcome of cervicobrachial pain that is treated conservatively, by Rasmussen et al. PMID- 12096244 TI - High mutation rate in the NS1 gene of parvovirus B19 DNA amplified from skeletal muscle of a case of mixed connective tissue disease. PMID- 12096245 TI - Treatment of AA amyloid with chlorambucil. PMID- 12096246 TI - Re: The role of ultrasonography in the diagnosis and management of idiopathic plantar fasciitis. PMID- 12096248 TI - Robin Goodfellow. PMID- 12096250 TI - Aortic clamping during elective operations for infrarenal disease: The influence of clamping time on renal function. AB - OBJECTIVE: Aortic clamping proximal to the renal arteries is sometimes necessitated during infrarenal and juxtarenal aortic surgery and may be associated with an increased risk of renal ischemia and its consequences. The aim of the study was to estimate this risk and possibly identify a "safe" duration of renal ischemia. METHODS: Medical records were retrospectively reviewed for 60 consecutive patients (from 1987 to 1994) with abdominal aortic aneurysm (n = 43) and occlusive disease (n = 17) confined to the infrarenal or juxtarenal aorta who underwent infrarenal aortic reconstruction with temporary suprarenal clamping. The data obtained included risk factors, preoperative and postoperative serum creatinine level, blood urea nitrogen (BUN) value, proteinuria before surgery, and suprarenal clamping times. RESULTS: The mean age of the patients was 64.4 years (+/- 11.4 years), and 74% were men. Concomitant cardiac disease was present in 41% of the patients, and 9% had diabetes. The preoperative creatinine level was 1.21 mg/dL (+/- 0.54 mg/dL), and the BUN value was 16.6 mg/dL (+/- 7.8 mg/dL). During surgery, blood flow to the renal arteries was interrupted for 32.0 minutes (+/- 17 minutes). None of the surviving patients needed dialysis or had signs of acute renal failure after the operations, but transient azotemia (rise in creatinine level) occurred in 23% of the patients. Risk factors for this condition were high preoperative creatinine values and hypotension during surgery, but the main determinant was total renal ischemia time. Odds ratios for such transient renal dysfunction showed as much as a 10-fold risk when suprarenal aortic clamping was greater than 50 minutes as compared with 30 minutes or less. CONCLUSION: Postoperative renal function impairment is rare in this group of patients. If suprarenal clamp duration (renal ischemia time) is brief, patients with normal preoperative creatinine levels exhibit no increase or a marginal increase in BUN or creatinine levels after surgery. Accordingly, suprarenal aortic clamping less than 50 minutes in this patient group appears safe and well tolerated. PMID- 12096249 TI - Prolonged administration of doxycycline in patients with small asymptomatic abdominal aortic aneurysms: report of a prospective (Phase II) multicenter study. AB - BACKGROUND: The primary purpose of this study was to evaluate compliance, side effects, and safety associated with prolonged administration of doxycycline in patients with small asymptomatic abdominal aortic aneurysms (AAAs). A secondary goal was to determine how treatment with doxycycline influences circulating levels of matrix metalloproteinase-9 (MMP-9) in this patient population. METHODS: Thirty-six patients with AAAs (30 men and 6 women; mean age, 69 +/- 1 years) were enrolled into a 6-month phase II study to evaluate treatment with doxycycline (100 mg orally twice a day). Aneurysm size was measured before and after treatment, and compliance and side effects were monitored. Plasma levels of doxycycline were measured midway through the study, and plasma MMP-9 concentrations were measured at baseline, 3 months, and 6 months. RESULTS: Thirty three of the 36 patients (92%) completed 6 months of doxycycline treatment. Significant treatment-related side effects occurred in five patients (13.9%), including three with cutaneous photosensitivity reactions (8.3%), one with tooth discoloration (2.8%), and one with yeast infection (2.8%). A high rate of compliance with treatment was seen, despite minor but frequent side effects, including nonspecific gastrointestinal symptoms (25%), easily managed episodes of photosensitivity (22.2%), and reversible tooth discoloration (5.5%). The mean plasma doxycycline level after 3 months was 4.62 +/- 0.68 ug/mL (median, 3.64 microg/mL; range, 1.31 to 14.39 microg/mL; n = 23 patients). No significant change was seen in AAA diameter (42.7 +/- 1.3 mm at 6 months versus 41.0 +/- 0.9 mm at baseline), and the overall rate of AAA expansion was 0.63% +/- 0.25% per month. The mean plasma MMP-9 level (n = 19 patients) was elevated at baseline (118.9 +/- 37.9 ng/mL; upper limit of normal, 85 ng/mL) but subsequently decreased to 83.8 +/- 32.9 ng/mL at 3 months (not significant versus baseline) and to 66.4 +/- 24.2 ng/mL at 6 months (P =.022 versus baseline). Only 21% of patients had an elevated level of plasma MMP-9 after 6 months of treatment compared with 47% at baseline (P <.05). CONCLUSION: Prolonged administration of doxycycline is safe and well tolerated by patients with small asymptomatic AAAs and is associated with a gradual reduction in plasma MMP-9 levels. Further studies are needed to evaluate the long-term effects of doxycycline on the rate and extent of aneurysm growth and the potential use of plasma MMP-9 levels as a biomarker of aneurysm disease progression. PMID- 12096251 TI - Small bowel motility and transit after aortic surgery. AB - OBJECTIVE: The inability to tolerate feedings after aortic surgery prolongs hospitalization. The aim of this study was to define jejunal manometric and small bowel transit characteristics associated with the ileus that follows transperitoneal aortic surgery. METHODS: Five male patients who underwent transperitoneal infrarenal aortobifemoral bypass had intraoperative placement of a jejunal multilumen catheter. The open abdomen allowed precise placement of pressure recording ports at 20, 22, 24, 26, 28, and 38 cm past the ligament of Treitz. Three-hour manometric studies were done after surgery and for 3 postoperative days. The migrating motor complex was identified visually on the manometric tracings, and pressure waves were identified with computer and a motility index calculated. Motility data were compared with healthy control data previously reported in the literature. Small bowel transit was determined with barium and serial abdominal radiographs. RESULTS: All patients had ileus develop with return of bowel sounds at 2 to 7 days (median, 6 days) and flatus at 3 to 9 days (median, 7 days) after surgery. Jejunal motor activity was present within 6 hours of surgery, but the motility index was less in patients then in control subjects. The postoperative migrating motor complexes differed from control subjects in having more phase I, less phase II, and more frequent phase IIIs. Phase III retrograde migration was common in the patients but not in the control subjects. Small bowel transit was 2 days or greater in all patients. CONCLUSION: Motor activity is present in the jejunum shortly after aortic surgery. However, the activity is decreased in intensity and the fasting cycle differs from control subjects. Retrograde migration of phase III is the most likely abnormality, resulting in delayed small bowel transit. The data would predict a high rate of enteral feeding intolerance early after surgery. Future studies should focus on pharmacologic manipulation to rapidly return small bowel motility to a more normal state after aortic surgery. PMID- 12096252 TI - Lipoprotein (a) in patients with aortic aneurysmal disease. AB - OBJECTIVE: Lipoprotein (a) is an independent risk factor for atherosclerosis. Atherosclerotic degeneration is usually found in abdominal aortic aneurysms (AAAs), whereas thoracic aortic aneurysms (TAAs) caused by aortic dissection are not suggested to be linked pathogenetically to atherosclerosis. Lipoprotein (a) was analyzed in patients with AAA and TAA and in healthy individuals in relation to the extent of atherosclerosis. METHODS: Included in the case control study were patients with AAA (n = 75) and TAA with dissection (n = 39) and healthy control subjects (n = 43), for a total of 157 participants. Serum lipoprotein (a) was measured with nephelometry. Lipoprotein (a) levels were compared between age matched and gender-matched paired samples of the three groups, and an association of lipoprotein (a), aortic aneurysm, and the extent of atherosclerosis was determined in multivariate analysis. RESULTS: Median lipoprotein (a) levels of patients with AAA and TAA and of control subjects were 18.9 mg/dL (interquartile range [IQR], <9.6 to 40.5), less than 9.6 mg/dL (IQR, <9.6 to 16.7), and less than 9.6 mg/dL (IQR, <9.6 to 16.3), respectively. Lipoprotein (a) was positively associated with the extent of atherosclerosis in patients and control subjects (P <.0001). Lipoprotein (a) levels of patients with AAA were significantly higher compared with patients with TAA (P <.0001) and control subjects (P <.0001). Multivariate analysis confirmed an independent association between lipoprotein (a) and AAA (P =.009). No significant differences of lipoprotein (a) were found between patients with TAA and control subjects (P =.3). CONCLUSION: The lipoprotein (a) serum level, an indicator of atherosclerosis, is significantly elevated in patients with abdominal aneurysms independently of cardiovascular risk factors and the extent of atherosclerosis. Patients with TAAs caused by dissection have lipoprotein (a) levels comparable with healthy individuals. PMID- 12096253 TI - Acute normovolemic hemodilution and intraoperative cell salvage in aortic surgery. AB - OBJECTIVE: The objective of this study was to report current transfusion requirements and outcomes in patients undergoing elective aortic surgery with autologous transfusion. METHODS: This was a retrospective review of transfusion practice in infrarenal aortic surgery in a tertiary vascular unit with a longstanding interest in autologous transfusion. One hundred and ten consecutive patients underwent infrarenal aortic surgery with a combination of acute normovolemic hemodilution (ANH) and intraoperative cell salvage (ICS). All patients underwent hemodilution to a target hemoglobin concentration of 11 g/dL and underwent ICS with a centrifugal device. RESULTS: Median blood loss was 1140 mL (interquartile range [IQR], 683 to 1609 mL) in 78 aneurysm repairs and 775 mL (IQR, 400 to 1225 mL) in 32 aortobifemoral bypasses for occlusive disease (P =.02), resulting in a median salvaged red cell volume of 403 mL (IQR, 256 to 563 mL) for aneurysm repairs and 250 mL (IQR, 200 to 290 mL) in bypass surgery (P =.001). Thirty-six patients (33%) needed transfusion of stored blood, for a total of 115 units, with just four patients needing more than five units. The mortality rate was 8% (9/110). With multivariate analysis, low hemoglobin level (P =.006) and low platelet count (P =.023) were associated with stored blood transfusion. CONCLUSION: Blood loss is too small to justify ICS in surgery for occlusive disease; ANH alone may be a suitable strategy. With appropriate experience, the combination of ANH and ICS may render crossmatching unnecessary, even in aortic aneurysm surgery. PMID- 12096254 TI - Higher prevalence of obstructive airway disease in patients with thoracic or abdominal aortic aneurysm. AB - BACKGROUND AND AIM: The risk factors of aortic aneurysm (AA) are comparable with those described for chronic obstructive pulmonary disease. This study was performed to determine whether patients with AA have a higher than average prevalence of obstructive airway disease. METHODS: We performed pulmonary function tests in 240 consecutive patients (182 men and 58 women; age, 70 +/- 10 years) with thoracic or abdominal AA. The results were compared with those in individuals without obvious cardiovascular disease (control) and in patients with coronary artery disease who were matched for age, gender, smoking status, and other atherosclerotic risk factors. RESULTS: Patients in the AA group had a lower forced expiratory volume in 1 second/forced vital capacity (%) and carbon monoxide diffusing capacity (%/predicted value) than did the control group (P <.01). The proportion of patients with airway obstruction, defined as forced expiratory volume in 1 second/forced vital capacity of 70% or less, was higher in the AA group (100/240; 42%) than in the control (51/223; 23%) and coronary artery disease (43/238; 18%) groups. Multiple logistic regression analysis results revealed that the presence of an AA and male gender were associated with a higher risk of airway obstruction (odds ratio, 2.928; 95% CI, 1.722 to 4.979; and odds ratio, 1.622; 95% CI, 1.055 to 2.493, respectively). CONCLUSION: These data suggest that AA may be a risk factor indicative of the presence of chronic obstructive pulmonary disease. A higher prevalence of depressed pulmonary function should be suspected as a preoperative risk in presence of thoracic or abdominal AA as compared with other types of cardiovascular disorders. PMID- 12096255 TI - Nonoperative management with selective delayed surgery for large abdominal aortic aneurysms in patients at high risk. AB - OBJECTIVE: An accepted fact is that abdominal aortic aneurysms (AAAs) larger than 5.5 cm should undergo elective repair. However, subsets of these patients have serious comorbid conditions, which greatly increase operative risk. This study evaluated the outcomes of periods of protracted nonoperative observational management with selective delayed surgery in patients at high risk with large infrarenal and pararenal AAAs. METHODS: Among 226 patients with AAAs more than 5.5 cm, we selected 72 with AAAs from 5.6 to 12.0 cm (mean, 7.0 cm) for periods of nonoperative management because of their prohibitive surgical risks. Comorbid factors included a low ejection fraction of 15% to 34% (mean, 22%) in 18 patients, 1 second forced expiratory volume less than 50% (mean, 38%) in 25, prior laparotomy in 10, and morbid obesity in 22. Follow-up examination was complete in the 72 patients for the 6 to 76 months (mean, 23 months) that they underwent nonoperative treatment. Fifty-three patients ultimately underwent operation because of AAA enlargement or onset of symptoms after 6 to 72 months (mean, 19 months) of nonoperative treatment. RESULTS: Of the 72 selected patients, 54 (75%) are living and 18 (25%) are dead. Seven patients who underwent only nonoperative treatment presently survive after 28 to 76 months (mean, 48 months). Of the 18 deaths, AAA rupture occurred in only three patients (4%) who were observed for 12, 31, and 72 months before rupture. Nine other deaths (13%) occurred after 6 to 72 months from comorbidities unrelated to the AAA. Six of the 53 patients who underwent delayed operation died within 30 days of operation (11% mortality rate). The mortality rate for the 154 good-risk patients with an AAA who underwent prompt open or endovascular repair was 2.2%. CONCLUSION: These data indicate that some patients with large AAAs and serious comorbidities are acceptably managed for long periods (6 to 76 months) with nonoperative means. Substantial delays of 12 to 76 months resulted in an AAA rupture rate of only 4%, and 13% of these patients (nine of 72) died of comorbidities unrelated to AAA rupture or surgery. Mortality rate in this group of patients, when operated, was 11% (six of 53). These findings support the selective use of nonoperative management in some patients with large AAAs and serious comorbidities. PMID- 12096256 TI - Subdural hematoma after thoracoabdominal aortic aneurysm repair: an underreported complication of spinal fluid drainage? AB - OBJECTIVE: Cerebrospinal fluid (CSF) drainage is a commonly used adjunct to thoracoabdominal aortic aneurysm (TAAA) repair that improves perioperative spinal cord perfusion and thereby decreases the incidence of paraplegia. To date, little data exist on possible complications, such as subdural hematoma caused by stretching and tearing of dural veins, should CSF drainage be excessive. We reviewed our experience with patients in whom postoperative subdural hematomas were detected. METHODS: The records of 230 patients who underwent TAAA repair at the Johns Hopkins Hospital between January 1992 and February 2001 were reviewed. RESULTS: Eight patients had subdural hematomas (3.5%). The four men and four women had a mean age of 60.6 years; two of these patients had a connective tissue disorder. All patients had lumbar drains placed before surgery, including one patient who underwent an emergency operation for rupture. Drains were set to allow drainage for CSF pressure greater than 5 cm H(2)O in all but one patient set for 10 cm H(2)O; spinal cooling was not performed in any patient. All drains were removed on the third postoperative day. In patients in whom subdural hematomas developed, the mean amount of CSF removed after surgery was 690 +/- 79 mL, which was significantly greater than the amount drained from patients in whom subdural hematomas did not develop (359 +/- 24 mL; P =.0013, Mann-Whitney U test). Six patients had postoperative subdural hematomas detected during hospitalization (mean postoperative day, 9.3; range, 2 to 16), and two patients were seen in delayed fashion after discharge from the hospital at 1.5 and 5 months. Four patients died of the subdural hematoma (50%); only one of these patients had neurosurgical intervention. All four survivors responded to neurosurgical intervention and are neurologically healthy. Two patients, both of whom were seen in delayed fashion, needed a lumbar blood patch. Multivariate logistic regression identified the volume of CSF drained as the only variable predictive of occurrence of subdural hematoma (P =.01). CONCLUSION: Subdural hematoma is an unusual and potentially catastrophic complication after TAAA repair. Prompt recognition and neurosurgical intervention is necessary for survival and recovery after acute presentation. Epidural placement of a blood patch is recommended if a chronic subdural hematoma is detected. Care should be taken to ensure that excessive CSF is not drained perioperatively, and higher (10 cm H(2)O) lumbar drain popoff pressures may be necessary together with meticulous monitoring of patient position and neurologic status. PMID- 12096257 TI - Management of cervical ribs and anomalous first ribs causing neurogenic thoracic outlet syndrome. AB - OBJECTIVE: Cervical and anomalous first ribs are rare conditions, occurring in less than 1% of the population. This manuscript reviews our management of neurogenic thoracic outlet syndrome (TOS) associated with these congenital anomalies. METHODS: During the past 26 years, 65 operations were performed for abnormal ribs that produced symptoms of TOS. Of these, 54 operations were for neurogenic TOS and are covered in this paper. Indications for surgery were disabling pain and paresthesia and failure to respond to conservative treatment. Surgical technique for neurogenic TOS was supraclavicular cervical rib resection and scalenectomy without first rib resection in 22 cases, supraclavicular cervical and first rib resection in 17 cases, supraclavicular excision of anomalous first ribs in five cases, and transaxillary anomalous first rib resections in two cases (total, 46 cases). Eight reoperations were performed for recurrent TOS in patients who previously had undergone cervical and first rib resections. RESULTS: Neck trauma was the cause of neurogenic symptoms in 80% of patients with cervical or anomalous first ribs. The surgical failure rate was 28% for 46 primary operations. A significant variable in results was the etiology of the symptoms. The failure rate for patients in whom symptoms developed after work related injuries or repetitive stress at work was 42%, and the failure rates for patients whose symptoms followed an auto accident or developed spontaneously were 26% and 18%, respectively. The failure rate in each etiology group also was affected by the operation performed. The failure rate for cervical rib resection without first rib resection in the work-related group was 75% compared with a failure rate of 38% in the non-work-related group. In contrast, when both cervical and first ribs were resected, the failure rate in the work-related group fell to 25% and in the non-work-related group to 20%. These failure rates for the work-related and non-work-related groups are similar to our failure rates in patients without cervical ribs. CONCLUSION: Surgery for neurogenic TOS in patients with cervical ribs should include both cervical and first rib resection. The presence of cervical or anomalous first ribs in patients with neurogenic TOS does not improve the success rate from surgery compared with patients without abnormal ribs. Neck trauma is the most common cause for neurogenic TOS in patients with abnormal ribs. Cervical and anomalous first ribs are the predisposing factors rather than the cause. PMID- 12096258 TI - Endoscopic thoracic sympathicotomy for Raynaud's phenomenon. AB - PURPOSE: For many years, thoracic sympathectomy via open surgery was not used to treat Raynaud's phenomenon because of the invasiveness of this procedure and the poor long-term outcomes associated with it. However, with the introduction of endoscopic surgery, thoracic sympathectomy (or sympathicotomy) has been performed by some surgeons as a less invasive surgical option for patients with Raynaud's phenomenon. The less invasive procedure has the possibility of emphasizing merits of sympathectomy. The purpose of this study was to reevaluate the efficacy of sympathicotomy for Raynaud's phenomenon with endoscopic technique and its range of applicability. METHODS: Between December 1992 and August 2001, endoscopic thoracic sympathicotomy (ETS) was performed in 28 patients with Raynaud's phenomenon (of a total of 502 patients with autonomic disorders who underwent ETS) at National Kanazawa Hospital. We considered indications for surgical treatment of Raynaud's phenomenon to include severe chronic symptoms or nonhealing digital ulceration refractory to intensive medical therapy. All patients were mailed a self-assessment questionnaire after surgery to determine the immediate and long-term results of the procedure. Data from both initial and long-term follow-up examinations were obtained. RESULTS: Fifty-four ETS procedures were performed in 28 patients. No operative mortality was seen, and no occurrence of major complications necessitated open surgery. Initial resolution or improvement of symptoms was achieved in 26 of 28 patients (92.9%). However, later in the postoperative period, symptoms recurred in 23 of 28 patients (82.1%), although no recurrence of digital ulceration was seen throughout our observation. At the final follow-up examination (median follow-up period, 62.5 months), 25 patients (89.3%) reported overall improvement of the frequency and severity of their symptoms. CONCLUSION: Despite the high rate of recurrence, ETS clearly produced a high rate of initial relief. ETS did indeed promote healing of digital ulcers, and the procedure shows potential for reducing the severity of refractory symptoms. We consider ETS to be the method of choice for treatment of severe or refractory Raynaud's phenomenon, and especially for Raynaud's involving digital ulcer, because of its safety and efficacy. PMID- 12096259 TI - Outcomes of carotid endarterectomy: how does the Australian state of New South Wales compare with international benchmarks? AB - OBJECTIVES: The objective of this study was to determine the outcomes of carotid endarterectomy (CEA) in New South Wales, Australia. METHODS: This state-wide prospective audit of all public and private hospitals during a 6-month period had main outcome measures of 30-day mortality, 30-day fatal stroke, 30-day nonfatal stroke, and 30-day combined stroke and mortality. RESULTS: Fifty-three of 54 eligible surgeons participated (response rate, 98%). The audit obtained inpatient data for 689 internal CEA procedures performed on 666 patients. Of 666 first or only procedures, 462 (69.4%) were performed on patients who were symptomatic and 204 (30.6%) on patients who were asymptomatic. After adjustment for cluster by surgeon, we determined a state-wide 30-day death rate of 1.2% (95% CI, 0.3% to 2.1%), a 30-day fatal or nonfatal stroke rate of 3.0% (95% CI, 1.6% to 4.4%; ipsilateral stroke, 2.6%), a 30-day nonfatal stroke rate of 2.3% (95% CI, 1.1% to 3.4%; ipsilateral stroke, 2.0%), and a 30-day combined stroke and death rate of 3.5% (95% CI, 2.1% to 4.9%). A significant linear relationship was found between patient preoperative hypertensive status and the combined 30-day stroke and death rate (trend odds ratio, 3.5; 95% CI, 1.6 to 7.7). No association was seen between any main outcome measures and hospital volume (<10 CEAs versus > or = 10 CEAs). Four patients underwent surgery outside guideline recommendations (CEA performed for either < or = 30% stenosis or occluded carotid artery). CONCLUSION: This first comprehensive state-wide audit confirms that Australian surgeons achieve 30 day outcomes comparable with international benchmarks. PMID- 12096260 TI - Early control of the distal internal carotid artery during endarterectomy: achievability and results. AB - PURPOSE: The purpose of this study was the report of the results of a consecutive series of carotid endarterectomy (CEA), performed by one surgeon and independently assessed by a neurologist, in which the achievability of selective early control of the distal internal carotid artery (ICA) was prospectively recorded. METHODS: All patients who underwent CEA by the surgical author between November 17, 1999, and November 15, 2001, were entered into a prospective study during which early selective exposure and clamping of the distal ICA were attempted (with initial avoidance of carotid bifurcation exposure and retraction), which allowed the remainder of the procedure (in patients without shunting) to be performed with ICA clamp protection and anticoagulation therapy. All the procedures were performed with local cervical block anesthesia, all the patients underwent a vein patch procedure, and, with one exception, cerebral angiography was not used. Major morbidity (stroke and myocardial infarction), mortality, and consecutiveness were independently verified by a neurologist. RESULTS: 148 consecutive CEAs were performed. In 142 cases (96%), the soft distal ICA could be initially isolated and controlled without dissection of the plaque bearing bulb bifurcation area (group A), and in the remaining six cases, the carotid bifurcation had to be exposed and retracted to allow ICA clamping (group B). A shunt was needed in 15 cases (14 in group A, one in group B), which left 128 cases (86%) in which the procedure could be performed with the ICA clamped and the brain theoretically protected from particulate plaque embolism. There were no strokes or deaths, one patient had temporary vertebrobasilar ischemia, one patient had a myocardial infarct, one patient had a temporary accessory nerve palsy, one patient returned to the operating room for release of a cervical hematoma, and 94% spent one postoperative night in the hospital. CONCLUSION: Early selective distal ICA control is highly achievable during CEA without apparently compromising clinical results. Its use is relevant when selective methods of shunting that do not need initial control of the common and external carotid arteries (eg, local cervical block anesthesia, electroencephalography/other monitoring) are used. Further evaluation with transcranial Doppler scan monitoring is suggested to substantiate the theoretic potential of this method in the reduction or elimination of particulate plaque embolism. PMID- 12096261 TI - Prevalence and progression of internal carotid artery stenosis in patients with peripheral arterial occlusive disease. AB - OBJECTIVE: The purpose of this study was to determine the prevalence of significant carotid stenosis, to identify risk factors increasing this prevalence, and to determine the risk of progression of stenosis, in patients with peripheral arterial occlusive disease who are neurologically asymptomatic. STUDY DESIGN: Consecutive patients who underwent evaluation in a vascular laboratory for peripheral arterial occlusive disease, who had no recent neurologic symptoms, were investigated. RESULTS: From July 1999 to December 2000, 620 patients underwent duplex scanning on one occasion, and 417 on two occasions. The average age was 72 +/- 10 years, and 61% were men. An occluded internal carotid artery was found in 4.8% of patients. The prevalence of a carotid stenosis >50% was 33% on the initial evaluation. Age of more than 70 years (P =.007), diabetes mellitus (P =.042), history of stroke (P =.011), and ankle/brachial index of less than 0.8 (P =.0006), were independently associated with carotid stenosis >50%. The odds ratio associated with each of these risk factors was similar. The prevalence of carotid stenosis >50% was 16%, 21%, 38%, 47%, and 44% for patients with no, one, two, three, and four risk factors, respectively. The highest prevalence of carotid stenosis >50% was identified in patients with ankle/brachial indices of less than 0.4 (59%). During the follow-up period, no patient had a cerebrovascular event. In 15% of carotid arteries, progression from one class of stenosis to a more severe class was observed, and 6.5% of patients progressed from a lower degree to 50% to 99% stenosis. No differences in progression of disease were identified when the variables of age, diabetes, previous stroke, and ankle/brachial index of less than 0.8 were studied or when patients with zero to two of these putative risk factors were compared with patients with three or four. CONCLUSION: Screening for carotid stenosis in asymptomatic patients with peripheral vascular disease is justifiable, but not mandatory, when two or more risk factors are present or when the ankle/brachial index is less than 0.4. Rates of progression to clinically significant stenosis are low and do not justify reevaluation every 6 months. Further research to identify the optimal interval for reevaluation is needed. PMID- 12096262 TI - Reproducibility of constant-load treadmill testing with various treadmill protocols and predictability of treadmill test results in patients with intermittent claudication. AB - BACKGROUND: Most trials on the reliability of constant-load treadmill testing use one pair of treadmill settings (speed, grade) only. The question of whether the results can be applied to tests with different settings is left open. Also, claudication distances measured with differing settings are not comparable, rendering the comparison of the results from different trials difficult. This study evaluates the reliability of constant-load testing with various workloads and compares them with claudication distances achieved with walking at normal speed on level ground but also evaluates whether metabolic equivalent (MET) normalization can be used to translate the results of different treadmill tests into each other. METHODS: Fifteen patients with claudication underwent repeated treadmill testing with different treadmill settings, including speeds of 2.0, 3.2, and 4.0 km/h (1.25, 2.0, and 2.5 mph, respectively) and grades of 0% and 12%. The walking capacity was also tested on level ground with a speed chosen by the individual patient. Results of virtual treadmill tests with all possible combinations of the speeds and the grades used were predicted from real tests, with MET normalization. The relationship between real and predicted claudication distances was tested with regression modeling. RESULTS: Reliability coefficients (RCs) for the absolute claudication distance (ACD) were superior to RCs for the initial claudication distance. RCs for ACD ranged between 0.61 and 0.95, with increasing values found with increasing workloads. The best coefficients for the regression of measured on predicted claudication distances were achieved with a model on the basis of a power function (r = 0.963). The model was only appropriate for the prediction of group mean results from clinical trials but cannot be applied to single patient data. For proof of concept, the model was tested with six published studies in which the claudication distances of a patient sample were double treadmill tested with different workloads. The result of the second test was predicted from the first test, and estimated and measured claudication distances were compared. The mean difference (all trials) was 7.9%, whereas the maximum difference amounted to 16.5%. CONCLUSION: For an optimal treadmill test reliability, higher workloads should be used and ACD should be preferred over initial claudication distance. MET normalization provides the basis for the comparability of treadmill test results achieved with different test conditions. PMID- 12096263 TI - The severity of muscle ischemia during intermittent claudication. AB - PURPOSE: The degree of ischemia during intermittent claudication is difficult to quantify. We evaluated calf muscle ischemia during exercise in patients with claudication with near infrared spectroscopy. METHODS: A Critikon Cerebral Redox Model 2001 (Johnson & Johnson Medical, Newport, Gwent, United Kingdom) was used to measure calf muscle deoxygenated hemoglobin (HHb), oxygenated hemoglobin (O(2)Hb), and total hemoglobin levels and oxygenation index (HbD; HbD = O(2)Hb - HHb) in 16 patients with claudication and in 14 control subjects before, during, and after walking on a treadmill for 1 minute (submaximal exercise). These measures were repeated after a second maximal exercise in patients with claudication and after 7 minutes walking in control subjects. Near-infrared spectroscopy readings during maximal exercise were then compared with a model of total ischemia induced with tourniquet in 16 young control subjects. RESULTS: Total hemoglobin level changed little during exercise in both patients with claudication and control subjects. HHb levels rose, and O(2)Hb level and HbD falls were more pronounced in patients with claudication than in control subjects after submaximal and maximal exercise. During maximal exercise, HbD fell markedly by a median (interquartile range) of 210.5 micromol/cm (108.2 to 337.0 micromol/cm) in patients with claudication compared with 66.0 micromol/cm (44.0 to 101.0 micromol/cm) in elderly control subjects and 41.0 micromol/cm (36.0 to 65.0 micromol/cm) in young control subjects (P <.001). This fall also was greater than the HbD fall induced with tourniquet ischemia at 90.8 micromol/cm (57.6 to 126.2 micromol/cm; P =.006). CONCLUSION: Hemoglobin desaturation in exercising calf muscle is profound in patients with claudication, considerably greater even than that induced with three minutes of tourniquet occlusion. Further studies are necessary to investigate the relationship between the inflammatory response and near-infrared spectroscopy during exercise in patients with claudication. PMID- 12096264 TI - The impact of walking impairment, cardiovascular risk factors, and comorbidity on quality of life in patients with intermittent claudication. AB - OBJECTIVE: The objective of this study was to assess the impact of walking impairment, cardiovascular risk factors, and comorbidity on quality of life (QOL) in patients with intermittent claudication (IC). MATERIAL AND METHODS: The prospective observational study was conducted in the setting of a vascular outpatient department of a teaching hospital. QOL was assessed in 200 consecutive patients with IC, with a reduced version of the World Health Organization Quality of Life Assessment Instrument-100. The reduced instrument assesses 17 facets of QOL within five domains (Physical and Psychological Health, Level of Independence, Social Relationships, and Environment). Age, gender, degree of IC, risk factors, comorbidity, as recommended by the Society for Vascular Surgery/North American Chapter of the International Society for Cardiovascular Surgery (SVS/ISCVS), and the presence of back, hip, or knee symptoms were analyzed as possible predictors of QOL. Multiple regression analyses were run with each of the QOL facets and domains as dependent variable. A probability value of less than.05 was considered to be statistically significant. RESULTS: Male gender was found to be a predictor of better scores for Energy and Fatigue and for Sleep and Rest. Women had more Negative Feelings. The presence of back, hip, or knee symptoms was a significant predictive value for many aspects of QOL. With more concomitant diseases, patients had lower scores on the facets of Overall QOL and General Health and of Energy and Fatigue and showed more dependence on medication and treatments. The degree of IC, as expressed in the SVS/ISCVS classification, was a statistically significant predictor of QOL on the domain Level of Independence and its facets Mobility, Activities of Daily Living, and Working Capacity and the facets Pain and Discomfort, Sexual Activity, and Transport. Hypertension was the second most important single predictor of QOL in patients with IC. CONCLUSION: QOL in patients with IC is only partially determined by the severity of walking limitation as expressed in the SVS/ISCVS classification. The significant impact of cardiovascular risk factors and comorbidity and the presence of back, hip, or knee symptoms on QOL should be recognized and taken into account in the treatment policy. PMID- 12096265 TI - The use of color-flow duplex scan for the detection of endoleaks. AB - OBJECTIVE: Computed tomographic (CT) scan represents the criterion standard for surveillance of endoleaks after endoluminal repair of abdominal aortic aneurysms (erAAAs). Given need for surveillance, risks, and expense of CT scan, the accuracy of color-flow duplex (CFD) scan after erAAA was determined. METHODS: During a 43-month period, patients enrolled in phase II and III of the AneuRx Multicenter Clinical Trial at our institution underwent CFD scan 1 month after erAAA. Patients with CFD scan results that were positive for endoleak underwent CT scanning at 3 months after erAAA, and those with CFD scan results that were negative for endoleak underwent CT scanning at 6 months after erAAA. RESULTS: Seven of 79 patients (9%) who underwent CFD and CT scanning had the diagnosis of endoleak. All endoleaks that were diagnosed with CT scan were detected with CFD scan. One patient had positive results for endoleak with CFD scan at 1 month and then negative results with CT scan at 3 months. Although this may represent resolution of endoleak, this case was counted as a false-positive result. When compared with CT scan, CFD scan had a sensitivity of 100%, specificity of 99%, positive predictive value of 88%, negative predictive value of 100%, and accuracy of 99%. CONCLUSION: CFD scan is an accurate test for the detection of endoleak after erAAA. In addition, most endoleaks diagnosed with CFD scan at 1 month continued to be present at 6 months. This important finding increases the emphasis on the use of this noninvasive test and may initiate earlier intervention of endoleak. PMID- 12096266 TI - Type II endoleaks: predictable, preventable, and sometimes treatable? AB - OBJECTIVE: The purpose of this study was to evaluate the effect of preoperative coil embolization of lumbar and inferior mesenteric arteries on the incidence of type II endoleak after endovascular abdominal aortic aneurysm repair. METHODS: The subjects were consecutive patients who underwent EVAR between January 1996 and January 2001. Patent aortic side branches were identified with preprocedural spiral computed tomographic scanning and calibrated angiography. Coil embolization was performed before EVAR. Patients were followed up with plain radiographs and ultrasound and dual phase spiral computed tomographic scans. Digital subtraction angiography was performed when endoleak was suspected. The outcome measures were the incidence of type II endoleaks and changes in maximum aortic sac diameter (Dmax). RESULTS: Forty patients underwent EVAR, with a median duration of follow-up of 24 months (range, 3 to 48 months). Before surgery, the inferior mesenteric artery was patent in 16 patients (45%) and the lumbar arteries in 21 patients (53%). Inferior mesenteric artery embolization was successful in 13 of 16 patients (81%). Lumbar embolization was attempted in 13 patients and was successful in eight (62%). During EVAR, successful sac exclusion was achieved in 38 patients (95%). None of the patients who underwent embolization before EVAR had type II endoleak develop, eight of 13 patients (62%) with patent lumbar arteries had endoleaks develop (P =.006), and three of these patients subsequently underwent successful coil embolization. Type II endoleak was associated with a 2.0-mm median increase in Dmax (P =.045). A 3.0-mm median reduction in Dmax was seen in the absence of type II endoleak (P =.002). CONCLUSION: Type II endoleaks are predictable, preventable, and sometimes treatable. Significant sac shrinkage occurs in the absence of lumbar endoleak but not in the presence of type II endoleak. PMID- 12096267 TI - Broadening the applicability of endovascular aneurysm repair: the use of iliac conduits. AB - OBJECTIVE: Suboptimal iliac anatomy has sometimes precluded endovascular repair of abdominal aortic aneurysm (AAA). In an effort to increase the applicability of endovascular repair, a limited retroperitoneal approach and iliac conduit was used in some patients with unsuitable iliac anatomy at high risk for open repair. METHODS: Charts and imaging studies of 312 patients who underwent endovascular (AAA) repair at the Cleveland Clinic Foundation between June 1999 and November 2000 were reviewed. Among these, 22 patients with complex iliac anatomy had an iliac conduit placed. Seventeen of these procedures were planned, but five were unplanned and placed after an iliac artery injury. A group of 17 patients who underwent a standard endovascular repair without conduits was selected and matched to the 17 patients in the planned conduit group by baseline comorbidities. The conduits were 8-mm or 10-mm polyester grafts sewn proximally to the common iliac artery and provided unobstructed access to the aneurysm. After insertion of the endograft device through the conduit, the distal end of the conduit was anastomosed to the external iliac or common femoral vessels. RESULTS: Operative time and estimated blood loss were higher among patients in whom conduits were performed, especially when performed urgently as an unplanned procedure. Although operative time and intensive care unit and hospital stays were longer for the group of patients with iliac conduits, the cardiac pulmonary and renal complication rates were similar for the conduit and the standard endovascular repair group. CONCLUSION: The use of a limited retroperitoneal approach and iliac conduit for patients with difficult iliac anatomy increases the applicability of the endovascular repair of AAA. This technique should be considered when an open surgical approach is inadvisable on the basis of medical comorbidities. PMID- 12096268 TI - Limb interventions in patients undergoing treatment with an unsupported bifurcated aortic endograft system: a review of the Phase II EVT Trial. AB - INTRODUCTION: Both supported and unsupported bifurcated endograft limbs develop flow-restricting lesions, including kinks, stenoses, and occlusions, which can be identified during or after surgery. Recognition and intervention are essential to achieve long-term graft patency and a satisfactory functional result. This report represents a comprehensive retrospective review of graft limb interventions from the Phase II EVT Trial with the Endovascular Grafting System unsupported bifurcated endograft (Guidant/EVT, Menlo Park, Calif). METHODS: The study population consists of 242 patients who underwent treatment with bifurcated endografts implanted during the EVT Phase II Trial. Graft limb interventions have been divided into two groups: those in whom the intervention occurred during surgery versus those in whom the intervention occurred after surgery. Parameters studied included type, incidence, and timing of graft limb intervention, indications for intervention, procedures performed, and overall patient outcome. RESULTS: The mean follow-up period was 31 months. Primary, primary assisted, and secondary limb patency rates were 61.6%, 93.7%, and 97.1%, respectively. Technical success rate at case completion was 97.5%. In 68 of the 242 cases, limb interventions were performed during surgery to assure patency (28.1%). In 28 cases, interventions were performed after surgery (11.6%). Of these postoperative limb problems, 82% occurred during the first 6 months. Repeat limb interventions were necessitated in three patients (1.2%). Within the intraoperative intervention group, perceived indications included kinks (15%), stenosis (57%), dissection (6%), graft redundancy (12%), and instances of twists, thrombosis, and pressure gradients (10%). These findings were successfully managed with percutaneous transluminal angioplasty only (41%), percutaneous transluminal angioplasty and stent (50%), and various combined interventions. Within the postoperative intervention group, symptomatic indications included stenosis (46%) and thrombosis/occlusion (54%). These postoperative limb events were successfully managed with stent (64%), thrombolysis (32%), and femoral-femoral bypass (21%). When limb dysfunction developed in the postoperative setting, it most often occurred within the first 6 months of implantation. Only one patient in this Phase II cohort had a lower extremity amputation unrelated to a graft limb abnormality. CONCLUSION: The unsupported bifurcated limbs of this endograft necessitated primary adjunctive intervention in 40% of cases. Primary intervention was two times more likely to be performed at the time of the implant rather than after surgery. Repeat limb interventions were not common. Endograft limb flow problems were successfully treated with standard endovascular or surgical interventions or both. These data may support prophylactic stenting of unsupported Ancure graft limbs. A strategy that includes both intraoperative and early postoperative graft limb surveillance is essential to detect reduced limb flow. PMID- 12096269 TI - Percutaneous coil embolization of postcatheterization arterial femoral pseudoaneurysms. AB - STUDY DESIGN: This study was a prospective monocentric study to assess the safety and effectiveness of percutaneous embolization with coils of postcatheterization femoral pseudoaneurysm (PCFP). PATIENTS AND METHODS: Seventeen PCFPs of 32-mm mean diameter in 16 patients were embolized while anticoagulant or antiplatelet therapy was maintained. Ultrasound scan-guided compression repair failed at least one time in 13 cases and was contraindicated in the four remaining cases. With ultrasound-Doppler scan guidance, the PCFPs were percutaneously punctured with a 16-gauge intravenous catheter. An angiogram was performed through the catheter to ensure its location within the sac. Stainless steel spring coils with synthetic fibers were introduced within the PCFP with fluoroscopic control. Successful thrombosis was checked with ultrasound-Doppler scan and was repeated at days 1, 30, and 180 when possible. RESULTS: All PCFPs of 32-mm mean diameter were successfully treated with two to nine coils. After embolization, gentle additional compression was necessary for complete occlusion, with a mean duration of 6.3 minutes (range, 0 to 15 minutes), except in one case with treatment with abciximab in which it was 45 minutes. All procedures were uneventful and painless. The mean follow-up period was 9.5 months (range, 1 to 21 months). Two recurrences (11.7%) were observed, and one was successfully treated with a second embolization. CONCLUSION: Percutaneous embolization with coils appears to be a safe and effective method for treatment of PCFP. It may be performed in patients undergoing anticoagulant or antiplatelet therapy and must be attempted when ultrasound scan-guided compression repair has failed or is contraindicated. PMID- 12096270 TI - The impact of the development of a program to reduce urgent (off-hours) venous duplex ultrasound scan studies. AB - OBJECTIVE: The purpose of this study was to develop a protocol for evaluation of patients suspected of having deep venous thrombosis (DVT) who came to the hospital after normal noninvasive vascular laboratory (NIVL) working hours in an attempt to both retain registered vascular therapists and provide excellent patient care. METHODS: We developed a protocol for evaluation of patients suspected of having DVT who were seen at night after normal NIVL working hours (8 am to 9 pm). After exclusion of those with contraindications, all eligible patients either received low-molecular weight heparin (LMWH; 1 mg/kg) and were sent home or were placed at bedrest in the emergency department until the NIVL opened the following morning. Those sent home were given maps to the NIVL and told what time to report. Parameters examined were numbers of after-hours venous duplex ultrasonography (VDU) studies, patient morbidity and mortality, and RVT retention. We compared results from October 1, 1997, through September 30, 1998, the year before the program started, with those from October 1, 1998, through September 30, 2000, the 2 years after program implementation. Data were compared with chi(2) analysis. RESULTS: During the study period, 161 patients suspected of having DVT were seen at the NIVL after hours and were deferred for study until the morning. Eighty-eight patients received prophylactic LMWH and were sent home, and 73 patients were placed in observation. All had VDUs the next morning at 8 am. No deaths, pulmonary emboli, or immediate complications of anticoagulation therapy were seen. After implementation, we noted an 89% reduction in after-hours VDU studies. Before program implementation, four RVTs resigned during an 18-month period because of issues relating to excessive call. From 1998 through 2000, the NIVL had 100% retention of its nine RVTs. CONCLUSION: The development of the LMWH program has reduced after-hours VDUs by 89% without causing patient morbidity or mortality. Retention of RVTs has been 100% since program implementation. PMID- 12096271 TI - Ambulatory venous pressure measurements: new parameters derived from a mathematic hemodynamic model. AB - PURPOSE: The mechanism of venous pressure decrease during exercise still remains unclear. To explore the components involved with the regulation of ambulatory venous pressure, we reinvestigated the pattern of pressure decrease during tiptoe exercise with a newly developed mathematic model. METHODS: Seventeen healthy limbs of 16 volunteers (normal group) and 35 limbs of 33 patients with signs and symptoms of chronic venous insufficiency were studied. Seventeen limbs had superficial venous incompetence (SVI), and 18 showed deep venous incompetence with or without concomitant superficial venous reflux. All subjects were examined with ambulatory venous pressure measurements. As parameters obtained from serial phasic changes in pressure during tiptoe movements, the pressure reduction fraction per step (decreasing component) and the pressure increase per step (increasing component) were calculated with application of the mathematic hemodynamic model and assessed comparatively in the three groups with different types of reflux (normal, SVI, and deep venous incompetence group). RESULTS: The pressure increase per step was significantly different in each of the three groups (P <.0001, with analysis of variance), whereas no apparent difference was seen in the mean pressure reduction fraction per step among the groups. With addition of the inflation of ankle cuff, the value of pressure increase in limbs with SVI was significantly reduced (P =.0004, with Wilcoxon signed rank test), although no changes were seen in the pressure reduction fraction in each group. CONCLUSION: Our results indicate that the pressure reduction fraction, representing calf muscle pump function, is independent of the existence or site of valve incompetence. On the other hand, the pressure increase, corresponding to the degree of reflux during exercise, correlates strongly with the severity of venous insufficiency. The theoretic model can separate the two components responsible for ambulatory venous pressure changes, calf muscle pump function and venous reflux, and provide better understanding of venous hemodynamics. PMID- 12096272 TI - Effect of endovascular stent strut geometry on vascular injury, myointimal hyperplasia, and restenosis. AB - PURPOSE: Early restenosis and the development of myointimal hyperplasia in stented blood vessels have been attributed to deep vascular injury with fracture of the internal elastic lamina (IEL). The purpose of this study was the evaluation of the vascular wall response to superficial injury (without IEL rupture) after balloon angioplasty and intravascular stent placement in porcine arteries and the determination of the effect of stent strut geometry on the degree of vessel injury and early restenosis. METHODS: Balloon-expandable stainless-steel stents were placed into the iliac arteries of 10 Sinclair miniature swine that had been fed an atherogenic diet. A Palmaz stent, with rectangular struts and smooth corners, was randomly assigned to one iliac artery (group 1), and a novel stent, which was designed and manufactured in the laboratory with thicker struts and sharper corners specifically to induce large wall stress concentrations, was placed in the contralateral iliac artery (group 2). Intravascular ultrasound scan was used in all deployments to ensure accurate balloon sizing and to avoid stent overexpansion and deep vascular injury. At 90 days after implantation, the animals were killed, the stented vessels harvested, and histomorphometric analysis performed. RESULTS: Deployment of novel stents in group 2 resulted in a statistically higher incidence rate of deep vascular injury (fracture of the IEL) compared with group 1, despite identical balloon size used for deployment (with Student t test, P <.05). Vessels with deep injury showed a 10-fold increase in myointimal thickening compared with those vessels in which the IEL remained intact. A statistically higher restenosis rate was observed for group 2 (33.5% +/- 19.90%) compared with group 1 (20.39% +/- 14.70%). For both stent designs, there was a trend toward lower degrees of restenosis within the mid-portion of the stent. For superficially injured arteries in both groups, no correlation was observed between the amount of vessel wall/medial layer compression and the development of restenosis from myointimal hyperplasia. CONCLUSION: Maintenance of an intact IEL is an important factor in the prevention of myointimal hyperplasia and restenosis in stented porcine iliac arteries. The alteration of stent strut height and geometry does not significantly affect restenosis and the development of myointimal hyperplasia in vessels with superficial injury. Superficial injury elicits a response that is independent of stent strut geometry and vessel wall compression. Stent strut profile may, however, increase local vessel wall stress concentrations, leading to IEL rupture and an exaggerated response to injury. PMID- 12096273 TI - Simvastatin inhibits human saphenous vein neointima formation via inhibition of smooth muscle cell proliferation and migration. AB - OBJECTIVE: Migration and proliferation of vascular smooth muscle cells (SMCs) contributes to intimal hyperplasia in saphenous vein (SV) bypass grafts, which leads to patency-threatening stenosis. Evidence for the involvement of basement membrane-degrading matrix metalloproteinases (MMPs) and growth factors in mediating SMC migration and proliferation has been presented in a number of in vitro and in vivo models. 3-Hydroxy-3 methylglutaryl CoA reductase inhibitors (statins) are widely used in patients with atherosclerosis and are claimed to have additional effects beyond cholesterol reduction. We therefore examined the effects of simvastatin, a commonly prescribed statin, on the proliferation and migration of cultured human SV SMC and on neointima formation and MMP activity in human SV organ cultures. To clarify its mode of action, we studied in parallel the effects of a specific MMP inhibitor, marimastat. STUDY DESIGN: Human SV specimens were obtained from patients who underwent coronary artery bypass grafting, and were cultured for 14 days in the presence of three concentrations of simvastatin and subsequently processed for measurement of MMP activity and neointimal thickness measurements. Cultured SV SMCs were used to construct growth curves in the presence of 10% fetal calf serum or 10% fetal calf serum supplemented with simvastatin or marimastat. Migration through a Matrigel basement-membrane matrix (invasion) was quantified with modified Boyden chambers. RESULTS: Simvastatin dose dependently reduced neointima formation (P =.004) in association with reduced MMP-9 activity (P =.03). SMC proliferation and invasion also were inhibited with simvastatin (P <.007 and P <.009, respectively). Marimastat dose dependently inhibited SMC invasion (P <.001) but importantly had no effect on SMC proliferation (P >.36). CONCLUSION: For effective control of neointimal development in vivo, a pharmacologic strategy should inhibit both SMC migration and proliferation. The ancillary properties of 3-Hydroxy-3 methylglutaryl CoA reductase inhibitors typified by simvastatin may be important in this regard. PMID- 12096274 TI - A 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, cerivastatin, suppresses production of matrix metalloproteinase-9 in human abdominal aortic aneurysm wall. AB - AIM: Abdominal aortic aneurysm (AAA) is a common vascular degenerative disease. AAA wall contains inflammatory cells that produce matrix metalloproteinases (MMPs) that probably contribute to elastolysis and remodeling of the aneurysm. 3 Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) have been shown to reduce the expression of various molecules (including MMPs) independently of their cholesterol-lowering effect. The aims of this study are to investigate whether statins could modulate the biology of AAA wall and have a potential therapeutic value against AAAs. METHODS: We performed immunohistochemical analysis, evaluated MMP-9 production in the aortic wall from patients with infrarenal AAA (n = 10) and control patients with aortoiliac occlusive disease (n = 8), and examined the effect of cerivastatin on MMP-9 production in the AAA wall with organ culture. RESULTS: Neutrophils and macrophages were the cellular sources of MMP-9 in the AAA wall. The tissue concentrations of both total and active MMP-9 were significantly higher in tissues from AAA walls than in control aortic walls. Cerivastatin (0.001 to 0.1 micromol/L) significantly reduced the tissue levels of both total and active MMP 9 in a concentration-dependent manner (P <.001), and the production of tissue inhibitor of MMP-1 was unaffected. Cerivastatin neither reduced the number of infiltrating neutrophils and macrophages nor enhanced apoptosis of those cells, as evaluated with terminal transferase-mediated deoxyurisine triphosphate nick end labeling. CONCLUSION: These results suggest that cerivastatin can directly modulate the biology of the AAA wall and suppress MMP-9 production in the AAA wall by inhibiting the activation of neutrophils and macrophages, indicating that statin therapy could be useful for the prevention or treatment of AAA. PMID- 12096275 TI - The effect of endothelial cell overexpression of plasminogen activator inhibitor 1 on smooth muscle cell migration. AB - INTRODUCTION: Plasminogen activator inhibitor-1 (PAI-1), a known inhibitor of plasminogen activators, may regulate smooth muscle cell migration (SMC) through alteration in matrix metalloproteinase (MMP) activity. METHODS: To study the effect of endothelial cell (EC) PAI-1 overexpression on SMC migration, RT-PCR was used to clone the full length PAI-1 gene, which was ligated into the pCMV/myc/ER expression vector. With electroporation, bovine aortic ECs were transfected with either the PAI-1 construct or the empty vector as control. EC PAI-1 overexpression was shown with a specific PAI-1 activity assay and enzyme-linked immunosorbent assay. The effect of EC PAI-1 overexpression on SMC migration was measured with a modified Boyden-chamber assay. SMC MMP expression was measured with zymography. RESULTS: Selected clones (EC9, EC21) had a three-fold to five fold increase in PAI-1 activity compared with untransfected EC and empty vector EC (ECC). Similarly, enzyme-linked immunosorbent assay results showed a 3.5-fold to 5.5-fold increase in PAI-1 levels in EC9 and EC21 versus ECC. Untransfected EC and ECC had similar effects on SMC migratory patterns. Migration of SMC exposed to PAI-1 overexpressing EC was inhibited by 35% to 57% compared with ECC. This inhibitory effect was reversed with addition of exogenous urokinase-type plasminogen activator (uPA). Zymography showed downregulation of MMP-2 and MMP-9 in SMCs exposed to PAI-1 overexpressing EC. CONCLUSION: PAI-1 overexpression with transfected EC inhibits SMC migration. This effect may be mediated through decreased SMC MMP activity. PMID- 12096276 TI - Correlation of a simple direct measurement of muscle pO(2) to a clinical ischemia index and histology in a rat model of chronic severe hindlimb ischemia. AB - PURPOSE: The lack of suitable experimental models of chronic severe limb ischemia and deficiencies in the available methods that allow for direct intermittent measurement of regional limb perfusion are obstacles to the evaluation of recently developed molecular strategies to reverse severe limb ischemia. Our aim was to develop a model of clinically relevant severe limb ischemia and correlate a simple direct measurement of muscle pO(2) to a clinical ischemia index, muscle mass, and capillary density. METHODS: Severe hindlimb ischemia was induced in 44 adult rats with ligation of the left common iliac artery, the femoral artery, and their branches. The effect of ischemia on muscle pO(2) was measured in the left gastrocnemius with room air and with 100% oxygen at 3, 10, 24, and 40 days after ischemia was induced. Clinical ischemia index, muscle mass, cellular proliferation, and capillary density also were assessed. RESULTS: The clinical ischemia index of the left limb was most severe at day 10, with evidence of pressure sores, a pale and dusky limb, and abnormal gait. With the rats breathing room air, muscle pO(2) was significantly lower in the left limbs than in the right limbs at days 3, 10, 24, and 40. After an oxygen challenge (100% O(2)), muscle pO(2) was significantly lower at 3, 10, and 40 days. At 3 days, the fraction of muscle mass per total body weight of the left tibialis anterior (TA) was significantly greater than the right TA as a result of edema and inflammation. By days 10, 24, and 40, the left gastrocnemius and TA masses were significantly less than the right as a result of muscle atrophy. Histopathology showed severe necrosis in the left gastrocnemius and TA on day 3. Inflammation was greatest by day 10. Necrotic muscle regenerated but remained atrophic at 40 days. The TA was slower to recover than the gastrocnemius. Capillary densities and capillary-to-muscle fiber ratios were greater in the ischemic limb than in the normal limb at day 24. Cellular proliferation as determined with bromodeoxyuridine labeling reagent staining was maximal in the ischemic limb at day 3. CONCLUSION: We have developed a rat model of chronic severe hindlimb ischemia with persistent ischemia as shown with a simple direct measurement of muscle pO(2) for up to 40 days. This model of severe hindlimb ischemia may be applicable for future studies of molecular strategies to treat severe limb ischemia in humans. PMID- 12096277 TI - Stent graft infection after abdominal aortic aneurysm repair: a case report. AB - A 77-year-old man had clinical and radiologic signs of graft infection develop 1 year after stent grafting for abdominal aortic aneurysm. Blood cultures grew Bacteroides fragilis, and cultures of the aneurysm sac grew Enterococcus. The patient's condition was successfully managed with staged extraanatomic revascularization followed by graft excision. Although stent graft infection to date is extremely rare, some aspects peculiar to the placement of these devices potentially could increase their susceptibility to infection. Recognition and standard techniques in management can lead to successful outcome. PMID- 12096278 TI - Delayed retrograde aortic dissection after endovascular stenting of the descending thoracic aorta. AB - A 66-year-old patient underwent emergency endovascular repair of a descending thoracic aneurysm because of suspected aortic rupture. Two weeks later, a small saccular aneurysm of the aortic arch was treated with open surgery. An unexpected intraoperative finding was retrograde dissection of the aortic arch and of the ascending aorta that was not seen on the postprocedural computed tomographic scans after endografting. The ascending aorta, the aortic arch, and the proximal part of the descending thoracic aorta were successfully replaced with a Dacron graft with deep hypothermia, circulatory arrest, and retrograde cerebral perfusion. Awareness that this life-threatening complication that necessitates extensive cardiovascular surgery can occur not only during or immediately after endovascular stenting of the thoracic aorta but also as much as several days or perhaps even weeks after the procedure is important. PMID- 12096279 TI - Successful endovascular therapy of a penetrating zone III internal carotid injury. AB - Penetrating injuries to the internal carotid artery in zone III of the neck can be a significant challenge to the operating surgeon. Direct surgical exposure and repair of the internal carotid artery at the skull base can be extremely difficult, and surgical options for treatment of a pseudoaneurysm at this location are limited. We present a case of an 18-year-old man who sustained a single gunshot wound to the distal cervical internal carotid artery that led to a pseudoaneurysm managed with endovascular exclusion. Recent literature on the surgical and endovascular management of distal carotid injuries is reviewed. PMID- 12096280 TI - Arterial tumor embolism caused by metastatic melanoma: case report and literature review. AB - Arterial tumor embolization is a rare complication of neoplasms. This report describes a 64-year-old female with metastatic melanoma and acute arterial embolization to the left lower extremity. With surgical embolectomy, a clot was retrieved that had an atypical bland solid lead point. Pathologic examination confirmed metastatic melanoma. Further echocardiography revealed an intracardiac mass consistent with metastatic disease. The patient was seen a month later with embolization to the right middle cerebral artery. PMID- 12096281 TI - A brief note on overlapping confidence intervals. AB - Clinical researchers frequently assess the statistical significance of the difference between two means by examining whether the two 95% confidence intervals overlap. The purpose of this brief communication is to illustrate that the 95% confidence intervals for two means can overlap and yet the two means can be statistically significantly different from one another at the alpha = 0.05 level. PMID- 12096282 TI - Regarding "Ambulatory venous pressure measurements: new parameters derived from a mathematic hemodynamic model". PMID- 12096283 TI - Complying with advance directives in the operating room. PMID- 12096284 TI - Regarding "Extracranial carotid aneurysm in Takayasu's arteritis". PMID- 12096286 TI - Regarding "Vein transposition in the forearm for autogenous hemodialysis access" and "Basilic vein transposition: an underused autologous alternative to prosthetic dialysis angioaccess". PMID- 12096288 TI - Desperately seeking an upside. PMID- 12096289 TI - Dyspnea: recognizing and managing an invisible problem. PMID- 12096290 TI - The healing power of presence: respite from the fear of abandonment. PMID- 12096291 TI - Illuminating problems, defining processes, and improving outcomes: the essence of oncology nursing research. PMID- 12096293 TI - Patients' perceptions of fatigue in response to biochemotherapy for metastatic melanoma: a preliminary study. AB - PURPOSE/OBJECTIVES: To explore patients' perceptions of fatigue in response to biochemotherapy treatment for metastatic melanoma. DESIGN: A descriptive correlational, cross-sectional study. SETTING: A cancer center in the midwestern United States. SAMPLE: 12 adult patients between the ages of 28-70 who received at least one cycle of biochemotherapy treatment for metastatic melanoma (stages III and IV) from the inpatient or outpatient services of a midwestern cancer center. METHODS: A demographic data sheet and the Revised Piper Fatigue Scale (PFS) were used to collect data at a single point in time after patients received at least one cycle of biochemotherapy. FINDINGS: The majority of patients who received biochemotherapy reported severe or moderate fatigue. Female patients' total fatigue scores were higher than those of male patients. Fatigue duration varied from hours to months, with a maximum duration of 12 months after biochemotherapy treatment. All of the patients reported that the most direct causes of their fatigue were metastatic melanoma and biochemotherapy treatment. CONCLUSIONS: Patients who received biochemotherapy treatment for metastatic melanoma reported moderate to severe fatigue. Female patients experienced more intense fatigue than male patients. The findings also supported the multidimensionality of fatigue construct identified in prior fatigue studies. The four dimensions/subscales of fatigue assessed by the Revised PFS were highly correlated to total fatigue scores. IMPLICATIONS FOR NURSING: Biochemotherapy is a newer treatment modality for metastatic melanoma. Fatigue, one of the severe toxicities from biochemotherapy treatment, necessitates attention from nurses. The findings will assist nurses in teaching patients about fatigue that may be expected during or after biochemotherapy and about self-care strategies to manage fatigue. PMID- 12096292 TI - Pain and fatigue management: results of a nursing randomized clinical trial. AB - PURPOSE/OBJECTIVES: Through a randomized clinical trial, to compare patients undergoing an initial course of chemotherapy who report pain and fatigue at baseline and who are receiving conventional care alone with those receiving conventional care plus a nursing intervention on outcomes reported at 20 weeks. SETTING: Chemotherapy clinics of two comprehensive and two community cancer centers. METHODS: Interviews were conducted at baseline and 10 and 20 weeks. An 18-week, 10-contact nursing intervention utilizing problem-solving approaches to symptom management and improving physical functioning and emotional health was implemented. SAMPLE: The sample consisted of 53 patients in the experimental arm and 60 in the control arm who reported pain and fatigue at baseline. VARIABLES: Pain and fatigue, numbers of other symptoms, and physical role impact and social functioning subscales from the Medical Outcomes Study 36 Short Form. FINDINGS: Patients who received the intervention reported a significant reduction in the number of symptoms experienced and improved physical and social functioning. Fewer patients in the experimental arm reported both pain and fatigue at 20 weeks. CONCLUSIONS: Behavioral interventions targeted to patients with pain and fatigue can reduce symptom burden, improve the quality of the daily life of patients, and demonstrate the "value-added" role of nursing care for patients undergoing chemotherapy. IMPLICATIONS FOR NURSING: These data support the "value added" role of nursing interventions for symptom management and improved quality of life during the course of cancer treatment. PMID- 12096295 TI - Interactive family learning following a cancer diagnosis. AB - PURPOSE/OBJECTIVES: To describe the experience of families when a member is diagnosed with cancer. DESIGN: Descriptive, qualitative study. SETTING: Patients' homes. SAMPLE: Eight adults, two to five months postdiagnosis, who were receiving radiotherapy or chemotherapy for stage I or II solid tumors and family members, including seven children between the ages of 13-18. Thirty people interviewed total. METHODS: Patients recruited from an oncology outpatient clinic and gynecologic inpatient unit of a teaching hospital interviewed on one occasion with at least two immediate family members in patients' homes. Semistructured interviews were tape-recorded and analyzed for themes and categories using the techniques of constant comparison. MAIN RESEARCH VARIABLES: Transitions from health to illness. FINDINGS: Families described a learning process in which information was gathered, interpreted, and shared. Families learned together by reviewing the past, gathering and sharing information, and sharing their experiences of living with someone undergoing treatment for cancer. By revealing their own personal perspectives, patients taught their families about their illness experiences and what constituted effective support. CONCLUSIONS: Interactive family learning is a mode of learning and a form of support in which the whole family may participate early in the process of learning to live with cancer. IMPLICATIONS FOR NURSING: Nurses can facilitate patient and family learning by considering the interactive manner in which families acquire information. By acknowledging how past experiences with cancer inform the present, nurses can help families identify beliefs influencing the illness experience. By including families in teaching sessions; facilitating communication between patients, families, physicians, and nurses; and providing take-home learning materials, nurses can facilitate shared information gathering. Nurses should acknowledge the value of learning about illness by experience, accept patients and families as experts, and encourage revelation of patients' and families' perspectives of the illness to enhance feedback on support and coping. PMID- 12096296 TI - Beginning treatment for childhood acute lymphoblastic leukemia: insights from the parents' perspective. AB - PURPOSE/OBJECTIVES: To report the perspective of parents during the initial stages of diagnosis and treatment for their children's acute lymphoblastic leukemia (ALL). DESIGN: Descriptive and phenomenologic. SETTING: Royal Children's Hospital, Brisbane, Australia. SAMPLE: Parents (mothers, n = 12; fathers, n = 4) of 12 children (ages 0-10) undergoing treatment for ALL. METHODS: Open-ended, audiotaped interviews were transcribed verbatim and analyzed. MAIN RESEARCH VARIABLES: Parents' experiences of their children's initial diagnosis and treatment of ALL. FINDINGS: The parents' insights provided a clear indication that the initial stage of treatment is highly stressful and parents may be overwhelmed by the experience. The situational stress translated into three potentially overwhelming emotional states: the stress of uncertainty, the shock of diagnosis, and a feeling of being trapped in an unpleasant emotional roller coaster ride. Honesty from healthcare professionals, the opportunity to share feelings, and an affirmation of the harshness of the situation were reported as helpful in dealing with the sense of being overwhelmed. CONCLUSION: Parents need honest information and sensitive emotional support to come to terms with the stresses associated with their children's diagnosis and initial treatment of ALL. Parents must have access to a safe environment where they can express their feelings and have the harsh reality of the initial stage of treatment affirmed. IMPLICATIONS FOR NURSING: Parents need considerable emotional support to negotiate the initial stage of treatment for ALL. Parents of children diagnosed with ALL must have honest information about diagnosis and treatment, as much information as possible for planning their daily lives, an opportunity to express real feelings, access to moments of personal space where feelings can be processed, and compassionate understanding of the emotional difficulties associated with this difficult life journey. PMID- 12096294 TI - Intervention to improve psychological functioning for newly diagnosed patients with cancer. AB - PURPOSE/OBJECTIVES: To test the effects of a computer-based nursing intervention designed to provide patients and family caregivers with concrete, objective information on symptom management; provide education about disease and treatment; coordinate medical resources; and provide emotional support and counseling. DESIGN: Two-site, randomized clinical trial. SETTINGS: A large, urban, midwestern, tertiary-cancer center and a community-based cancer center in a medium-sized midwestern city. SAMPLE: 109 patients newly diagnosed with breast, colon, or lung cancer who were receiving chemotherapy; 54 received standard care, and 55 participated in the intervention group. METHODS: Outcome data were collected via structured telephone interviews at three time points: baseline, midway through the intervention, and one month postintervention. The intervention consisting of nine visits, five in person and four by telephone, was conducted over 18 weeks by advanced practice oncology nurses. MAIN RESEARCH VARIABLES: Psychosocial functioning, anxiety, and depression. FINDINGS: Patients who received the intervention had significantly less depression between baseline and the midway point, as well as less anxiety and greater improvement in the role emotional and mental health subscales of the Medical Outcomes Study 36 Short Form. CONCLUSIONS: Cancer-care nursing interventions can decrease psychosocial morbidity and improve quality of life for newly diagnosed patients with cancer undergoing treatment. Additional research is needed to understand who benefited most from the intervention. IMPLICATIONS FOR NURSING: This nurse-directed intervention resulted in improved mental health for patients; however, physical subscales were not changed. Further work is needed to determine why depression and mental health were affected yet physical health and symptoms did not differ between groups. Results support the important role of nurses in addressing mental health issues in patients and families experiencing cancer. PMID- 12096298 TI - Addressing the support needs of women at high risk for breast cancer: evidence based care by advanced practice nurses. AB - PURPOSE/OBJECTIVES: To identify support needs of women at high risk for breast cancer and enhance an evidence-based service. DESIGN: Descriptive study. SETTING: A comprehensive, breast-health service for high-risk women. SAMPLE: 97 high-risk women with a 1.66% or greater five-year risk of breast cancer, atypical hyperplasia, lobular carcinoma in situ, or positive genetic screen. METHODS: A self-assessment questionnaire completed previsit and a satisfaction survey completed postvisit. MAIN RESEARCH VARIABLES: Women's perceived informational, emotional, and decisional support needs, current self-care practices, and satisfaction with the service provided. FINDINGS: Women under age 50 (n = 54) wanted information on breast cancer screening, risk of breast cancer, lifestyle options to lower risk, and hormone replacement therapy; older women (n = 43) wanted information on risk of breast cancer, lifestyle options, breast cancer screening, and chemoprevention. More than 75% of all women wanted information to help them make decisions on breast cancer prevention options, benefits, and risks. The satisfaction survey (N = 61) revealed that most women's needs were met. CONCLUSIONS: Support needs were consistent with the literature that focused primarily on younger women seeking genetic counseling. Proactive planning assisted with addressing the needs of these women. IMPLICATIONS FOR NURSING: A previsit questionnaire facilitates individualized proactive planning before the visit. However, further assessment of self-care practices and emotional needs is required. Interventions should evaluate outcomes, such as accurate risk perception, lifestyle changes, screening follow-through, and decision quality. Advanced practice nurses require specialized skills, including evidence-based risk communication, behavior modification, and decision support. PMID- 12096297 TI - Effect of caregiver perception of preparedness on burden in an oncology population. AB - PURPOSE/OBJECTIVES: To identify the level of lay caregiver burden and perception of preparedness that exists for the oncology population prior to and just after hospital discharge. DESIGN: Descriptive, correlational design using repeated measures. SETTING: Large, tertiary care, private medical center in the midwestern United States. SAMPLE: 59 lay caregivers of patients with cancer who were discharged from inpatient hospital status. METHODS: Caregivers completed Robinson's Caregiver Strain Index and Archbold's Preparedness Scale questionnaires immediately prior to discharge, 7-10 days postdischarge, and 28-30 days postdischarge. MAIN RESEARCH VARIABLES: Caregivers' perception of preparedness and burden. FINDINGS: Preparedness levels ranged from "somewhat" to "pretty well" prepared and were consistent over time. Burden levels were moderate and also consistent over time. Increased levels of preparedness were associated with decreased levels of burden, and that relationship was consistent over time. CONCLUSIONS: Caregivers' levels of preparedness can be enhanced, although no significant differences were reported in the period from prehospital discharge to one month following discharge. Burden experiences of caregivers are real and not affected by the transition from hospital to home. Burden experiences can be influenced by perceptions of preparedness. IMPLICATIONS FOR NURSING: Efforts to enhance the preparedness of lay caregivers can influence their burden experience. PMID- 12096300 TI - Musculoskeletal injuries among ERCP endoscopists in Canada. AB - BACKGROUND: There are few reports in the literature describing musculoskeletal complaints among endoscopists, and none are specific to those who perform endoscopic retrograde cholangiopancreatography (ERCP). PURPOSE: To examine the current practices of ERCP endoscopists and the prevalence of musculoskeletal injuries. METHODS: A self-report survey was sent to physicians practising ERCP across Canada identified through a pre-existing database. A second mailing was sent six weeks later to those who did not respond to the first mailing. RESULTS: Of 162 surveys sent, 122 responses were received, with five respondents indicating that they no longer performed ERCP and three declining to participate. Of the 114 participants, 67% reported at least one musculoskeletal complaint, and 58% reported two or more complaints. Seventy-four per cent attributed their symptoms to endoscopy and/or ERCP, and 79% reported that their condition was aggravated by performing ERCP. The most frequently reported pain symptoms were back pain (57%), neck pain (46%) and hand pain (33%), which are all consistent with the physical risks involved in performing ERCP. Only 51% reported taking regular breaks, and only 25% reported having fluoroscopy tables with adjustable heights. The room designs of the respondents' ERCP facilities were analyzed for ergonomic considerations: 67% had poor ergonomics and 33% had good ergonomics. Sixty-four per cent reported that they were interested in learning preventive strategies. CONCLUSIONS: Physicians who perform ERCP develop musculoskeletal injuries and are interested in learning about risk factor modification. PMID- 12096299 TI - Curative treatment for esophageal cancer: Vancouver Island Cancer Centre experience from 1993 to 1998. AB - OBJECTIVES: To review outcomes after curative treatment for esophageal cancer in the Vancouver Island Cancer Centre from 1993 to 1998. Curative treatments included esophagectomy alone, and chemoradiotherapy with "selective surgery" for patients with post-treatment-positive endoscopic biopsy or less than 75% regression on computed axial tomography scan, or with resectable local recurrence. METHODS: Patients undergoing esophagectomy alone, or primary chemoradiotherapy and "selective surgery" were reviewed. This was a retrospective, nonrandomized, institutional experience. Surgical complication, relief of dysphagia, disease-specific survival rates and prognostic factors were analyzed. RESULTS: Nineteen patients underwent esophagectomy alone. A total of 56 patients underwent primary chemoradiotherapy, of whom 16 had "selective surgery". Relief of dysphagia was similar in both groups of esophagectomy patients. Exploration for "selective surgery" was performed in 12 patients after their first postchemo-radiotherapy endoscopy (two patients had unresectable disease), and in seven for relapse, one of whom died intraoperatively. Overall, the mortality rate due to surgery was 3%. Chemoradiotherapy was not associated with more frequent serious surgical complications. For patients who underwent esophagectomy alone and those who underwent chemoradiotherapy plus selective surgery, the median survival times were 12.9 and 16.4 months, respectively, and the three-year survival rates were 21% and 37%, respectively. Seventeen of 25 patients who underwent chemoradiotherapy and who survived more than two years have not required selective surgery. For the two groups of patients combined, no single prognostic factor for survival was significant in multivariate analysis, but for patients who underwent chemoradiotherapy plus selective surgery, negative endoscopic biopsy was highly significant. CONCLUSIONS: Surgical complication and disease-specific survival rates after primary chemoradiotherapy with selective surgery compare favourably with esophagectomy alone in the curative treatment of esophageal cancer. A prospective, randomized trial is necessary for the definitive evaluation of the strategy of chemoradiotherapy and selective surgery. PMID- 12096301 TI - Giant liposarcoma of the esophagus. AB - Liposarcomas of the gastrointestinal tract are exceedingly rare. Only nine cases of esophageal involvement have been described. A 68-year-old woman presented with an episode of vomiting followed by extrusion of a polypoid mass from the mouth. This 10th case of esophageal liposarcoma is the first in the literature to report a recurrence 25 years after the first episode. PMID- 12096302 TI - Cholestasis: the ABCs of cellular mechanisms for impaired bile secretion- transporters and genes. AB - The transport of bile salts, organic anions and cations, bilirubin and other substances from the portal blood into the biliary system is accomplished through the action of an array of transporter proteins in the hepatocyte. Transporters on the basolateral membrane, which faces the space of Disse, are responsible for the uptake of bile salts and organic anions. Once translocated through the hepatocyte to the canalicular membrane, other ATP pumps provide the energy to export bile salts, phospholipids and organic ions into the bile. Canalicular transport is rate limiting. Defects in specific canalicular transporters are responsible for many of the intrahepatic cholestatic syndromes that occur in children and adults. Moreover, cholestasis provokes changes in several transport mechanisms, many of which appear to be compensatory and serve to protect the liver from the toxic effects of accumulated materials. The identification and characterization of the major transporters responsible for bile formation have yielded a more precise classification of the cholestatic syndromes of infancy and childhood, and are unlocking the molecular mechanism of acquired cholestasis in adults. This review identifies the basic physiology of bile production and the actions of the key transporters, indicates the clinical relevance and possible treatments of transport disorders, and provides an illustrative case. PMID- 12096303 TI - Photodynamic therapy for cancer: principles. AB - The principles of photodynamic therapy (PDT), using drugs (photosensitizers) that are activated by light to become cytotoxic, provide the basis for understanding the current and potential future clinical applications in gastroenterology, general oncology and other specialties. The properties of photosensitizers are key to their biological efficacy, while lasers and optical fibres allow convenient and flexible light delivery for endoscopic use. PDT has several distinct and unique advantages, both as a stand-alone treatment and in combination with other established modalities. The current limitations are also recognized, as is the need for rigorous randomized trials of this emerging technology. The fluorescence of many photosensitizers may be useful, either for (endoscopic) diagnosis or for PDT treatment guidance and monitoring. PMID- 12096304 TI - Who will pay for treatments administered in outpatient units? PMID- 12096305 TI - Abstracts from the annual general meeting of the Canadian Society for Clinical Nutrition. Nutrition in health and disease: the state of the nation. PMID- 12096306 TI - Apoptosis in liver disease. PMID- 12096307 TI - Calculation of esophageal variceal wall tension by ultrasonic microprobe and noninvasive pressure measurements. AB - OBJECTIVE: To calculate esophageal variceal wall tension by sonographic and noninvasive pressure measurements and to study the role of esophageal variceal wall tension in predicting the likelihood of esophageal variceal rupture. METHODS: In 28 patients with esophageal varices, a 20 MHz ultrasonographic transducer was used to image esophageal varices; the radius and wall thickness of the varices were calculated. Esophageal variceal pressure measurements were obtained noninvasively. The correlation between the esophageal wall tension and esophageal variceal pressure gradient were studied. RESULTS: There was a linear correlation between the esophageal wall tension and the variceal pressure gradient after curve fitting (r=0.87, p < 0.05). CONCLUSION: Esophageal variceal wall tension is an important parameter in predicting variceal rupture. PMID- 12096308 TI - [Comparative study of accidents and complications of laparoscopic cholecystectomy in cirrhotic and non cirrhotic patients]. AB - Among a total of 9000 laparoscopic cholecystectomies, 60 (0.66%) were performed in cirrhotics patients. We have analyzed the presence of intra- and postoperative complications and have made a statistical comparison between cirrhotics and the group of 8940 non-cirrhotics. Cirrhosis was associated with a higher rate of conversions, of intra- and postoperative complications and mortality. The particularities of the intervention in cirrhotic patients are evaluated. Data analysis shows that cirrhotic patients have special problems regarding operation indication, surgical technique and pre- and postoperative treatment. PMID- 12096309 TI - The pathogenesis of chronic alcoholic pancreatitis. AB - The pathogenesis of chronic alcoholic pancreatitis has not been elucidated yet. There are many theories about this topic. The first regards the initial injuries in the ductal pancreatic system. The second considers that the primary lesion is located in the acinar cells. Three hypotheses are elaborated in this respect: the intervention of toxic metabolites, oxidative stress or neuroimmmune system. The third theory sustains that a severe initial attack of acute pancreatitis may induce lesions typical of chronic pancreatitis. Recently the intervention of genetic factors, responsible for the susceptibility chronic pancreatitis has been put forward. PMID- 12096310 TI - Current trends of management for fissure in ano. AB - There are many options to treat chronic fissures in ano. Some of them are non surgical while the others are surgical. The efficacy claimed by each of the prevalent method is very high but the inconsistencies and contraindications are equally strong. To date, lateral sphincterotomy has been favoured by most of the proctologists, because it is the least extensive surgical procedure and is offering a long lasting relief in sphincter spasm. About 10 different techniques are presented in brief with their efficacy and fallacy. A new method combining the age-old technique of Lord's manual dilatation followed by radio surgery is also highlighted. The addition of radio surgery is found useful for refreshing the edges of the fissure and to tackle pathologies namely sentinel pile, small internal piles or hypertrophied anal papillae often found associated with chronic fissures. Revisiting the trends of treatment of chronic anal fissures, the most preferred options are the manual dilatation with radio surgery and the subcutaneous lateral anal sphincterotomy. Both methods are easy to perform, have negligible complications and no special setup is needed, except the radio surgical unit, in case of the first procedure. However, all the options should be presented to the patient with complete information about the method, cure rates, complications, and reversibility of the complications and the final choice should be left in his hands. PMID- 12096311 TI - Endoscopic clips -- what are they good for? AB - Several methods have been developed for the treatment of upper gastrointestinal bleedings, a condition associated with high mortality rates. In recent years new studies have been published reporting the use of endoscopic clips mainly for digestive bleeding. Our paper reviews recent publications, focusing mainly on present indications, such as: digestive bleeding, conservative treatment of perforations of the digestive tract, repair of anastomotic leaks, marking of digestive tumours, endoprosthestesis anchoring and assisting papilla cannulation. PMID- 12096312 TI - Endoscopic mucosal resection for early gastric cancer. A case report. AB - European experience in endoscopic mucosal resection (EMR) for early gastric cancer is still relatively low, since early stomach cancer is diagnosed at a much lower rate in Europe than in Japan and generally operable patients are referred to surgery for radical resection. Endoscopic mucosal resection or mucosectomy was developed as a promising technology to diagnose and treat mucosal lesions in the esophagus, stomach and colon. In contrast to surgical resection, EMR allows "early cancers" to be removed with a minimal cost, morbidity and mortality. We present the case of a patient with hepatic cirrhosis incidentally diagnosed with an elevated-type IIa early gastric cancer. Echoendoscopy was performed in order to assess the depth of invasion into the gastric wall confirming the only mucosal involvement. We performed an EMR using "cup and suction" method. After the procedure, the patient experienced an acute upper gastrointestinal bleeding from the ulcer bed requiring argon plasma coagulation. The histopathological examination confirmed an early cancer, without involvement of muscularis mucosae. The patient has had an uneventful evolution being well at six months after the procedure PMID- 12096313 TI - Preoperative "two-way" enteroscopy, followed by intraoperative enteroscopy in a patient with obscure-overt gastrointestinal bleeding and severe iron-deficiency anemia. AB - We present the case of a 69-year-old woman admitted to hospital because of chronic gastrointestinal bleeding of an unknown source with a consequent severe iron deficiency anemia (IDA), undiagnosed for the past 25 years. In the last three years the episodes of severe bleeding became frequent, usually followed by melena. The patient was admitted 11 times in different departments without the identification of the bleeding source. During the evolution of the disease, the biological exams showed a severe IDA with low values of hemoglobin, low serum iron, mixed deficiency depicted by bone-marrow examination, and a reticulocyte crisis after parenterally administered iron. Repeated upper (6) and lower (2) gastrointestinal endoscopies failed to find a source of bleeding. Push enteroscopy allowed the visualization of approximately 40 cm of the proximal jejunum, after the Treitz angle, and demonstrated multiple punctiform jejunal angiodysplasias, which bled excessively after bipolar coagulation. We also performed a total colonoscopy with intubation of the ileo-cecal valve and visualization of the terminal ileum on approximately 30 cm, without any pathological findings. Because endoscopic treatment was ineffective, we decided to perform a segmentary enterectomy, with the length of small bowel resection tailored by intraoperative enteroscopy. A favourable evolution after limited resection of the small bowel indicated the importance of both preoperative "two way" enteroscopy associated with intraoperative enteroscopy for diagnosing and treating the source of obscure gastrointestinal bleeding PMID- 12096314 TI - Laparoscopic total colectomy for asymptomatic familial adenomatous polyposis. AB - A 20-year-old female patient with familial adenomatous polyposis diagnosed on the occasion of the screening of her family, which had a history of three deaths due malignant colic polyposis, was submitted to total colectomy by laparoscopic technique. Though the histopathological examination performed on two polypi excised by endoscopic technique did not evidence signs of malignant shift, total colectomy was performed, observing the oncological principles. The intervention was uneventful and postoperative course uncomplicated, the patient being socially reintegrated after three weeks. Follow-up examinations performed up to 18 months after the operation showed a favourable evolution and no development in the size and number of polypi. PMID- 12096315 TI - Metastasis of malignant melanoma in the small intestine. A case report. AB - Malignant melanoma is the most common metastatic tumor of the gastrointestinal tract and can present with fairly non-specific symptoms. A 63-year old man with previous enucleation of the left eye for a malignant coroidian melanoma presented signs of intestinal subocclusion. Weight loss and a palpable mass deep on the paraumbilical left region were the significant physical signs. Because the state of the patient had worsened, the diagnosis was made by ultrasonography examination. Segmental intestinal resection with regional lymph node dissection was performed. Surgical resection can be performed safely as patients for whom all sites of disease are completely resected experience significant improvements in survival times, compared with patients who undergo an incomplete resection. For selected patients, surgical treatment of metastatic melanoma involving the gastrointestinal tract is appropriate therapy. PMID- 12096316 TI - Splenic abscesses. AB - Splenic abscesses are rare entities (autopsy incidence between 0.14-0.7%). The most frequent etiology is the septic emboli seeding from bacterial endocarditis (about 20% of cases) or other septic foci (typhoid fever, malaria, urinary tract infections, osteomielitis, otitis). The treatment of splenic abscesses was until recently splenectomy with antibiotherapy. The actual trends are more conservative (mini invasive or non-invasive) because the immunologic role of the spleen has been better understood over the last year PMID- 12096317 TI - Scintigraphic detection and localization of gastrointestinal bleeding. AB - By frequency and often through the difficulties of diagnosis and treatment, gastrointestinal (GI) bleeding plays an important role in the activity of medico surgical departments. Successful management of patient with GI bleeding depends on accurate localization of the bleeding site. The early etiological diagnosis of GI bleeding is important for the early optimal treatment (medical or surgical). In addition to the clinical exams, endoscopy and angiography may be used as invasive methods to localize the bleeding site. Beside these, the isotopic technique in detecting GI bleeding is non invasive, rapid and with high sensitivity. Correctly indicated and performed, the imaging methods, invasive or not, play an important role in the success of treatment. For the clinicians it is important to know the best algorithm of examination and the role of each technique and, bearing in mind the technical equipment of the hospital to use quickly the appropriate method rapidly. PMID- 12096318 TI - [Quiz of gastroenterology and hepatology]. PMID- 12096319 TI - The role of imaging for the evaluation of pancreatic cancer resectability. PMID- 12096321 TI - Laparoscopic colorectal surgery: an analysis of 36 cases. PMID- 12096323 TI - Is there any relationship between periodontitis and Helicobacter pylori infection? PMID- 12096324 TI - [Science and ethics]. AB - In current ethics debates, the protection of animals and the environment are central themes. In this context, reference is made to "ethics of responsibility". In the reality of scientific life and the conduct of science, "ethics of responsibility" have only had a modest impact. Reflection based on the history of thought may explain this finding. Upon abolishing God, man has redirected an initially religious love and devotion toward himself. The postulate of unfettered dominion over the Earth has become the supreme principle. "Theoria" in the sense of its original meaning, that is a celebratory admiration for sacred Nature, is incomprehensible to modern man. Nature has been stripped of its sanctity and transformed into something to be exploited for its human utility. Consequently, today the credo applies that to interpret the freedom of science according to ethical principles is utterly untenable. The author pleads for reconsideration. PMID- 12096325 TI - [Experimental animal research in the EU legislation]. AB - The legislation of the European Union captures experimental animal research only in part. Binding standards can be found in various legal records of the community legislation, and in particular in the so-called guideline for animal experimentation 86/609/EWG. But these guidelines do not represent an actual animal protection measure but rather one of harmonisation with the primary goal of unification of the regulations of the participating states in order to prevent distortions of competition and trade barriers which could harm the common market. Although the guideline contains some practicable approaches in the direction of an up-to-date animal experimentation law, it only defines general goals which allows for considerable leeway in national implementation within the individual EU countries, and it has only a limited area of legal operation. On the one hand, only vertebrae are being included, and on the other, the law is only being applied in the area of applied research and protects only animals used in product and substance-development or test procedures as well as those used in the framework of environmental protection. Various important fields of research are thus not subject to a common regulation and are assigned to national regulation. This concerns animal experimentation in education and training or for military or so-called defence-relevant medical purposes and, in particular, the whole area of basic research including the field of genetic engineering in animals with it's growing significance. The guideline is in need of widening it's scope of application as well as of various adjustments to recent scientific findings and developments in order to become suitable as a more restrictive animal protection law on the community level. It could also be desirable to include animal protection into the catalogue of community-goals in order to make it an independent component of the politics of the Union and to establish the groundwork for a decree of comprehensive and efficient regulations. PMID- 12096326 TI - [White Paper of the European Commission--strategy for a new chemicals policy]. AB - The current chemicals regulation in EU member states provides sufficient data on new chemicals but almost non on the about 100.000 existing chemicals. To improve the situation the EU Commission is proposing in a recent White Paper on a strategy for a new chemicals policy to regulate new and existing chemicals in an identical manner. The Commission suggests to use in vitro methods to provide information rapidly on hazardous health and environmental properties of chemicals. The fundamental change in the testing strategy, which is relying on in vitro methods rather than testing in animals is most welcome not only from the scientific and economic point of view but also from the perspective of the protection of animals and the environment. A calculation of the costs and time for testing all existing chemicals in animal experiments shows that this task is unrealistic from a financial point of view and also when taking into account the time required for testing. Taking into account the proposals provided by several working groups of the EU validation centre ECVAM during the past year, a concept is described that will allow the testing of all existing chemicals applying only non-animal tests. Since the new testing strategy will be cheaper and faster it will be more acceptable from both the financial and the ethical point of view. Finally, establishing the proposed new in vitro tests in the safety testing strategy will require considerable funding of validation studies by the EU Commission. PMID- 12096327 TI - The new EU chemicals policy--Discussions on details relevant for animal welfare. AB - The European Commission is planning to put forward drafts for a new chemicals legislation by June 2002. In fulfillment of an Environmental Council Conclusion, Working Groups have been set up for consultation during the ongoing preparatory stage. There, members of the General Directorates Environment and Enterprise discuss relevant topics with representatives from authorities, industry, environmental and animal protection organisations. There is agreement that animal tests shall be reduced to a minimum. However it is still unclear how this goal can best be achieved. In this context, the designing of testing strategies will play a major role. It is explained, why fixed test catalogues should be replaced by flexible tiered testing strategies and how concrete waiving strategies can contribute to avoiding animal tests. Another important aspect is the EU-wide implementation of a clause on the avoidance of duplicate testing, which is already enforced in Germany and Austria. In these Member States, first parties have to provide data from previously performed animal tests to second parties. Finally, it is discussed that the application of new non-animal tests can be promoted, if the revised EU chemicals policy once again contains the legal framework for an EU-specific acceptance of new test methods. PMID- 12096328 TI - Ecotoxicological assessment of surface waters: a modular approach integrating in vitro methods. AB - Today ecotoxicological evaluations of surface water quality are either based on field surveys or online biomonitoring, whereas the ecotoxicological quality of wastewater is mostly determined with standardised acute toxicity tests. In this paper we present a concept for the ecotoxicological evaluation of surface waters, where mainly in vitro tests are used for the screening of water samples, presenting the first tier of a two-tiered approach. In this first tier a battery of fast and cost-efficient test-systems are used as an early warning system. Thereby, the toxic potential of water samples will be identified. This modular approach allows the exchange or addition of test-systems if necessary. If a toxic potential is identified in a water sample, this sample can be investigated more thoroughly in a second tier where organisms are used. In this paper we focus mainly on the general approach and the description of the first tier. PMID- 12096329 TI - DarT: The embryo test with the Zebrafish Danio rerio--a general model in ecotoxicology and toxicology. AB - The acute fish test is an animal test whose ecotoxicological relevance is worthy of discussion. The primary aim of protection in ecotoxicology is the population and not the individual. Furthermore the concentration of pollutants in the environment is normally not in the lethal range. Therefore the acute fish test covers solely the situation after chemical spills. Nevertheless, acute fish toxicity data still belong to the base set used for the assessment of chemicals. The embryo test with the zebrafish Danio rerio (DarT) is recommended as a substitute for the acute fish test. For validation an international laboratory comparison test was carried out. A summary of the results is presented in this paper. Based on the promising results of testing chemicals and waste water the test design was validated by the DIN-working group "7.6 Fischei-Test". A normed test guideline for testing waste water with fish is available. The test duration is short (48 h) and within the test different toxicological endpoints can be examined. Endpoints from the embryo test are suitable for QSAR-studies. Besides the use in ecotoxicology the introduction as a toxicological model was investigated. Disturbance of pigmentation and effects on the frequency of heart beat were examined. A further important application is testing of teratogenic chemicals. Based on the results DarT could be a screening test within preclinical studies. PMID- 12096330 TI - [PCR and ELISA--in vitro alternatives to the mouse-bioassay for assessing the botulinum-neurotoxin-C1 production potential in environmental samples?]. AB - Botulism is one of the most important bird diseases world-wide and is caused by the intoxication with Botulinum-Neurotoxin-C1 (BoNt-C1), which is produced by toxigenic clostridia under appropriate conditions. Avian botulism leads regularly to large losses among the migrating bird populations breeding and resting at the saltwater pools of the Austrian national park Neusiedler See-Seewinkel. Despite of its ethical dubiousness and its high technical expense the mouse-bioassay is still used as the routine standard method for the detection of BoNt-C1. According to the 3R-concept, in vitro alternative methods for the qualitative detection of BoNt-C1 (immunostick-ELISA) and a corresponding BoNt-C1 gene fragment (nested PCR) were established. In order to estimate the BoNt-C1 production potential the methods were tested with sediment samples from different saltwater pools subjected to cultivation conditions appropriate for in vitro BoNt-C1-production. With the mouse-bioassay, 52 out of 77 samples were found to have a positive toxin production potential. The immunostick-ELISA showed a similar sensitivity as the mouse-bioassay and exhibited a highly significant positive correlation (r=0.94; p<0.001) with the mouse-bioassay in detecting BoNt-C1. The nested-PCR approach revealed higher numbers of positive BoNt-C1 gene fragment detections as compared to the direct toxin analysis approaches. A weak correlation (r=0.21; p=0.07) with the mouse-bioassay was discernible, no correlation was found with the immunostick ELISA (r=0.09; p=0.46). Obviously, the PCR approach detected the BoNt-C1 gene fragment in some of the samples where no toxin expression has occurred. Thus it is suggested that the qualitative immunostick-ELISA represents a potential in vitro alternative to the mouse-bioassay for assessing the BoNt-C1 production potential in environmental samples. In contrast, qualitative BoNt-C1 gene fragment detection via PCR led to an overestimation of the actual toxin production potential. PMID- 12096331 TI - [Improving the embryonic stem cell test (EST) by establishing molecular endpoints of tissue specific development using murine embryonic stem cells (D3 cells)]. AB - Blastocyst-derived pluripotent embryonic stem (ES) cells of the mouse can be induced to differentiate in culture into a variety of cell types, including cardiac muscle cells. In the embryonic stem cell test (EST) the capacity of ES cells of the mouse cell line D3 to differentiate into contracting cardiomyocytes is used to assess the embryotoxic potential of test compounds and in addition, the effects on the viability of ES cells and differentiated mouse fibroblasts (cell line 3T3) are compared. The three endpoints are used to classify the embryotoxic potential of chemicals after 10 days of exposure: (i) the inhibition of differentiation of ES cells into cardiomyocytes (ID50) and (ii) the decrease of viability of 3T3 cells (IC503T3) and (iii) ES cells (IC50D3) in a MTT cytotoxicity test. Applying linear analysis of discriminance, a biostatistical prediction model (PM) was developed to assign test chemicals to three classes of embryotoxicity. In an international validation study funded by ECVAM it could be demonstrated that the EST can predict the embryotoxic potential of a test compound as good as frequently used mammalian systems based on pregnant animals. In a joint project with major German pharmaceutical companies we are attempting to improve the EST by establishing molecular endpoints of differentiation (e.g. cardiac, neuronal, chondrogenic) in cultured ES cells. We have studied the expression of tissue specific proteins in ES cell cultures in the presence of embryotoxic chemicals by immunofluorescent antibody techniques, e.g. FACS analysis. The other groups are focusing on endogenous gene expression in early development by RT-PCR methods or the DNA microarray technique. The results obtained recently using molecular markers specific for cardiac differentiation and employing intracellular flow cytometry for quantification will be presented. Molecular endpoints will allow improvement of the EST by measuring gene expression patterns in a small number of murine ES cells. PMID- 12096333 TI - Comparative study of rabbit pyrogen test and human whole blood assay on human serum albumin. AB - A comparative study of rabbit pyrogen test and human whole blood assay was performed on released preparations of human serum albumin. In addition, the samples were spiked with 5 IU/ml (in whole blood 0.5 IU/ml too) and 10 IU endotoxin/ml. The unspiked samples were negative in both assays. The human whole blood test resulted in the same level of security for the products as the rabbit pyrogen test did. Both, the borderline 5 IU/kg and the 10 IU/kg-Spike partially lead to results of the rabbit test which would cause further testing with additional animals. In contrast, the human whole blood assay resulted in a 100% detection for the 5 IU/ml and 10 IU/ml-Spike. We designed a study protocol for a minimised number of test animals and were able to show the general usefulness of the human whole blood assay. PMID- 12096332 TI - Immortalisation of ovarian granulosa and Theca cells of the marmoset monkey Calllithrix jacchus. AB - In view of the increasing need for laboratory primates in biomedical research it is desirable to develop appropriate primate-specific cell culture models that could prevent or significantly reduce the increasing use of primary cultures and experiments with living animals. Follicular granulosa and theca cells are essential for the control of hormone-dependent processes such as the ovarian cycle and pregnancy, but also for the occurrence of hormone-dependent diseases. For this reason it is of great interest to know more about control mechanisms existing in these follicular cell types and the effect of pharmacological or toxicological agents on them. An immortalisation protocol for the two ovarian cell types of the marmoset monkey (Callithrix jacchus) has been developed. All cell lines established so far were examined with regard to the maintenance of known, tissue-specific features (e.g. hormone responsiveness and enzyme expression). The results obtained indicate that it is worth while cloning and characterising the cell lines in more detail so that they could be used after an adequate validation as a defined test system both for basic research as well as for pharmacological or toxicological screening. PMID- 12096334 TI - [Replacement of mouse and rat antibody production test; comparison of sensitivity between the in vitro and in vivo methods]. AB - Bacteria and viruses may be transmitted through contaminated biological materials such as transplantable tumors, cell lines, sera or other biological material. Currently, the screening of biological material is being done using the mouse or rat antibody production (MAP/RAP) test (serological testing). We decided to test and validate an alternative assay using polymerase chain reaction (PCR / RT-PCR) technology to detect viral contamination directly in biological material. The aim of this study therefore is the validation of our new PCR assays and the comparison of PCR and MAP test. For 6/14 viruses the conventional PCR, seems to be more sensitive and more specific in detecting murine viruses. In 12/14 viruses the RT-PCR is more sensitive than MAP-test. In 2/14 cases all detection methods have the same sensitivity. Furthermore, PCR screening clearly contributes to the principles of 3R as a replacement technique as it eliminates the need for using animals to screen for murine viruses in biological material. PMID- 12096335 TI - Enhancing foot skin blood flux in peripheral vascular disease using intermittent pneumatic compression: controlled study on claudicants and grafted arteriopaths. AB - Intermittent pneumatic leg compression (IPC) increases arterial calf inflow and foot skin blood flux in normal subjects and claudicants. Our hypothesis was that IPC could enhance foot skin blood flux after infrainguinal grafting and thus promote distal perfusion in limbs with tissue loss. The aim of this study was to compare the effects of three IPC modes [applied to the foot (IPCfoot), the calf (IPCcalf), or both (IPCfoot+calf)] on foot skin perfusion in healthy individuals, claudicants, and patients after infrainguinal arterial revascularization performed for critical or subcritical limb ischemia. Altogether, 20 healthy limbs, 22 claudicating limbs, and 36 limbs of arteriopaths with prior successful autologous femoropopliteal and femorodistal (18 each) grafts were examined. Five minute laser Doppler recordings were obtained from the pulp of the big toe in the sitting position, at rest, and during random applications of IPCfoot, IPCcalf, and IPCfoot+calf delivered at 120 mmHg for 4 seconds three times per minute. Foot skin blood flux increased using all IPC modes (p <0.001), with IPCfoot and IPCfoot+calf generating higher flux levels than IPCcalf (p <0.01) in all groups. Intergroup differences of flux with each of the three IPC modes were not significant. IPCfoot and IPCfoot+calf similarly (p > 0.14) produced a higher percentage flux increase than IPCcalf in all groups (p <0.004). Controls had a higher percentage flux increase with both IPCcalf and IPCfoot than did claudicants (p? 0.016). No differences were documented between normal and grafted limbs (p > 0.05). The percentage flux increase with IPCfoot+calf and IPCcalf was significantly higher in femorodistal grafts than in femoropopliteal ones (p ? 0.026). IPC enhances skin blood flux in limbs with infrainguinal bypass, claudication, and normal arteries, with IPCfoot and IPCfoot+calf being more effective than IPCcalf. Our findings suggest that IPC may be beneficial in limbs with impaired distal perfusion and thus may have clinical implications in the treatment of leg ulcers either prior to or after revascularization. PMID- 12096336 TI - Novel gain of function activity of p53 mutants: activation of the dUTPase gene expression leading to resistance to 5-fluorouracil. AB - Mutated forms of p53 are often expressed in a variety of human tumors. In addition to loss of function of the p53 tumor suppressor, mutant p53s contribute to malignant process by acquisition of novel functions that enhance transformed properties of cells and resistance to anticancer therapy in vitro, and increase tumorigenecity, invasiveness and metastatic ability in vivo. Searching for genes that change expression in response to p53 gain of function mutants may give a clue to the mechanisms underlying their oncogenic effects. Recently by subtraction hybridization cloning we found that the dUTPase gene is transcriptionally upregulated in p53-null mouse fibroblasts expressing the exogenous human tumor-derived His175 p53 mutant. Here we show that conditional expression of His175 and Trp248 hot-spot p53 mutants in p53-negative mouse 10(1) fibroblasts and human SK-OV3 and H1299 tumor cells results in increase in dUTPase gene transcription, an important marker predicting the efficacy of cancer therapy with fluoropyrimidine drugs. Using tetracycline-regulated retroviral vectors for conditional expression of p53 mutants, we found that transcription of the dUTPase gene is increased within 24 h after tetracycline withdrawal, and the cells acquire higher resistance to 5-FU. Additional inactivation of the N-terminal transcription activation domain of mutant p53 (substitutions in amino-acid residues 22 and 23) results in abrogation of both induction of dUTPase transcripts and 5-FU resistance. PMID- 12096337 TI - p38 MAPK-mediated activation of NF-kappaB by the RhoGEF domain of Bcr. AB - The oncogenic fusion protein p210 Bcr-Abl is causally associated with virtually all cases of chronic myelogenous leukemia. The wild-type Bcr product has several recognizable structural and functional motifs including a domain that contains guanine nucleotide exchange activity for Rho family GTPases (DH/PH domain). Although this domain is retained within p210 Bcr-Abl, it has no known signaling activities in vivo. Here we report that a fragment of Bcr that encodes the isolated DH/PH domain is a potent activator of the NF-kappaB transcription factor. Within the context of full length Bcr, this activity is regulated by proximal flanking sequences that suppress the DH/PH domain encoded guanine nucleotide exchange activity. NF-kappaB activation by Bcr is not mediated by nuclear translocation, but rather by p38 mitogen-activated protein kinase (MAPK) dependent modification of the RelA/p65 transactivation domain. Although we were able to demonstrate that Bcr can function as an exchange factor for Cdc42 in vivo, NF-kappaB activation appears to occur via a Cdc42-independent mechanism. These studies constitute direct evidence that the Bcr RhoGEF domain can function in vivo, and identify a new signaling activity that may contribute to the transforming potential of p210 Bcr-Abl. PMID- 12096338 TI - Adenovirus encoding HIV-1 Vpr activates caspase 9 and induces apoptotic cell death in both p53 positive and negative human tumor cell lines. AB - The targeted delivery of genes whose products arrest the cell cycle and/or induce apoptosis represent an important tool for the understanding and controlling forms of unregulated cell growth. The vpr gene product of HIV-1 has been reported to interfere with cell growth and induce apoptosis, but the mechanism of its action is not clearly understood. In order to study these important properties of Vpr, we created a recombinant adenovirus H5.010CMV-vpr (adCMV-vpr) as a tool to deliver the vpr gene to various cell lines to examine its biology. Vpr protein expression was confirmed by Western blot analysis in adCMV-vpr infected cells. We tested the effects of adCMV-vpr on cell growth of several tumor cell lines. Infection of both p53 positive and p53 deficient tumor cell lines with adCMV-vpr resulted in dramatic induction of cell death in short-term assays. We observed that apoptosis was induced through the mitochondrial pathway as we observed changes in the cytochrome c content accompanied by caspase 9 activation. As Bcl-2 is reported to interfere with apoptosis through the mitochondrial pathway, we examined the effect of adCMV-vpr in Bcl-2 over expressing cell lines. We observed that Bcl-2 overexpression does not inhibit adCMV-vpr induced apoptosis. The properties of adCMV-vpr inducing apoptosis through caspase 9 in a p53 pathway independent manner suggest that this is an important reagent. Such a vector may give insight into approaches designed to limit the growth of pathogenic human cells. PMID- 12096339 TI - Ectopic expression of cyclin E in estrogen responsive cells abrogates antiestrogen mediated growth arrest. AB - Estrogens stimulate proliferation of estrogen receptor positive MCF7 breast cancer cells while antiestrogens signal a G0/G1 growth arrest. In MCF7 cells, arrest is mediated through the CDK inhibitors p21 and p27 and through a decrease in cyclin E/CDK2 kinase activity. We found that in MCF7 cells, overexpression of cyclin E partially abrogates a tamoxifen mediated growth arrest. Overexpression of cyclin E is accompanied by a decrease in the levels of RB and CDK inhibitor p21 but an increase in CDK inhibitor p27. Cyclin E overexpression also alters the composition of E2F transcription factor complexes. The E2F4/p107/cyclin E/CDK2 complex, a minor component in proliferating control cells that is absent in growth-arrested cells, is more abundant in both proliferating and tamoxifen treated cyclin E overexpressing cells. Conversely, levels of the quiescence associated E2F/p130 complex is not detected in these cells. Expression from the E2F dependant promoter is elevated in proliferating and tamoxifen treated cyclin E overexpressing cells. This study suggests that a modest overexpression of cyclin E abrogates the tamoxifen mediated growth arrest through modification of the RB/E2F pathway. Moreover, these results provide one explanation of why some cells that express the estrogen receptor may be unresponsive to antiestrogens. PMID- 12096340 TI - Tumor formation in mice with somatic inactivation of the retinoblastoma gene in interphotoreceptor retinol binding protein-expressing cells. AB - The retinoblastoma suppressor gene product Rb has been assigned a critical role in cell cycle regulation, the induction of differentiation, and inhibition of oncogenic transformation. Inheritance of a mutant RB allele in humans is responsible for bilateral retinoblastoma, a malignant tumor of the retina. Trilateral retinoblastoma (TRB) is a rare variant of familial retinoblastoma in which, in addition to retinal tumors, tumors develop from the pineal gland, an organ ontologically related to the retina. Germline inactivation of Rb in mice leads to mid-gestational lethality with defects in erythropoeisis and neurogenesis. This embryonic lethality prohibits the analysis of Rb function in selected cell types at later stages of development or in the adult. Here, we describe the Cre-LoxP mediated somatic inactivation of Rb in a subset of neuroendocrine cells, including photoreceptor cells. We observed neuroendocrine tumors of the pineal and pituitary gland. These tumors invariably showed inactivation of Rb and Trp53. Remarkably, loss of Rb in photoreceptor cells does not lead to retinoblastoma or any phenotypic changes, not even when photoreceptor cells are made deficient in Rb, p107 and Trp53. Our results highlight the important differences that exist in tumor susceptibility between mice and man (e.g pineal gland) and question the photoreceptor cell origin of human retinoblastoma. PMID- 12096341 TI - Novel colon cancer cell lines leading to better understanding of the diversity of respective primary cancers. AB - A major obstacle to obtaining more detailed insights into the diversity of phenotypic and molecular changes occurring in colon cancer cells is the lack of low-passage colon cancer cell lines, which would still closely reflect the phenotype of the colon cancer cells in vivo. Here, we characterize eight novel, low passage number human colon carcinoma cell lines, originating from colorectal cancers extensively characterized in the clinics. All cell lines closely resemble the original tumors with respect to phenotype, markers and detectable genetic changes. Cell morphology and marker expression is highly variable, ranging from fully polarized cells correctly expressing all basolateral epithelial markers, to cells with mesenchymal characteristics and a complete loss of polarity due to delocalization or loss of junction complex proteins. The alterations in phenotype and epithelial marker expression correspond to changes already detectable in the primary tumor in vivo. Seven of the cell lines show chromosomal instability, while one cell line is characterized by microsatellite instability. p53 associated with K-ras mutations were detected in three cell lines. Hitherto non described E-cadherin mutations were found at both alleles in one cell line whereas in another cell line the E-cadherin protein was down-regulated. A stabilizing beta-catenin mutation (S45F) appears in the same cell line that carried the mutated E-cadherin gene. Six cell lines carried APC mutations, which in five of the lines led to an activated beta-catenin/Tcf/LEF signaling pathway. In accordance with beta-catenin/Tcf/LEF activation, the cell lines show increased migration and invasiveness. Our results show that the characterized, low-passage cell lines mirror the diversity of the individual tumors from which they were derived. Through molecular analyses of these cell lines we demonstrate that tumorgenicity events are much more diverse in human colon cancer than expected, despite the common origin of the tumors from a small patient group with similar tumor grading and clinical prognosis. PMID- 12096342 TI - Identification and characterization of the Myb-inducible promoter of the chicken adenosine receptor 2B gene. AB - Numerous studies have shown that the retroviral oncogene v-myb encodes a transcription factor (v-Myb) which interferes with the differentiation program of myelomonocytic cells. It is generally thought that v-Myb deregulates the expression of specific target genes and thereby causes transformation of these cells. By using an estrogen-inducible version of v-Myb we have previously identified the gene for the chicken A2B adenosine receptor (A2B-AR), a member of the seven-pass transmembrane receptor superfamily, as a bona fide target gene for v-Myb. The chicken A2B-AR gene is expressed in v-myb transformed myeloblasts as well as in c-myb expressing erythroblasts, offering the opportunity to study how Myb transcription factors activate a target gene in two different hematopoietic lineages. Here, we report the characterization of the promoter of the A2B-AR gene. We show that the A2B-AR promoter region contains an exceptionally large number of Myb binding sites, many of which contribute to the Myb-inducibility of the promoter. The same sites were required for promoter activity in myelomonocytic and erythroid cells. In contrast to the promoters of other Myb target genes the A2B-AR promoter was not activated synergistically by Myb and other lineage-specific transcription factors that have been identified as Myb cooperation partners before. Taken together, our data suggest that the activation of the A2B-AR promoter by Myb depends on the simultaneous binding of a large number of Myb molecules. PMID- 12096343 TI - hBUB1 defects in leukemia and lymphoma cells. AB - Tumorigenesis is a multi-step process involving a series of changes of cellular genes. Most solid tumors and hematopoietic malignancies often show abnormal chromosome numbers, the aneuploidy. The chromosomal aneuploidy keeps cells in the state of chromosomal instability (CIN) that will increase the mutation rate of cells affected and thus push them deeper into the process of tumorigenesis. The yeast genetic studies showed that normal distribution of chromosome during mitosis is under the surveillance of a set of genes, the spindle assembly checkpoint genes, that include the BUB and MAD gene groups and MPS. In some colorectal cancers with CIN it was found to have hBUB1 gene mutated and the mutated gene functions dominantly. We have examined a series of breast cancer cell lines with or without CIN for the hBUB1 gene mutation and found none. However, we detected various degrees of deletion in the coding sequences of the hBUB1 gene in cells from T lymphoblastic leukemia cell lines, Molt3 and Molt4, and cells from some acute lymphoblastic leukemia and Hodgkin's lymphoma patients. So far the lesions of deletion are in the kinetochore localization domain of the hBUB1 gene that may explain why the deletion lesions in the BUB1 gene cause aneuploidy in lymphoma and lymphoma cells. The deletions are heterozygous in nature. Like the mutated hBUB1 gene in colorectal cancer, the mutant hBUB1 cDNA from lymphoblastic leukemia cells behaves dominantly. PMID- 12096344 TI - The cyclin D1 high and cyclin E high subgroups of breast cancer: separate pathways in tumorogenesis based on pattern of genetic aberrations and inactivation of the pRb node. AB - In an attempt to identify subtypes of breast cancer and pinpoint patterns of cell cycle regulatory defects associated with clinical behaviour, proliferation and other transformation associated events, a multitude of cell cycle regulatory proteins were analysed in a material of 113 primary breast cancers. Increased proliferation was observed in two different scenarios; (1) with high cyclin D1 and elevated retinoblastoma protein (pRb) phosphorylation, (cyclin D1(high) tumours) or (2) with high cyclin E protein but low cyclin D1 and lack of corresponding pRb phosphorylation (cyclin E(high) tumours) indicative of an interrupted pRb pathway. Characteristic for cyclin E(high) tumours were further defects in p53, p27 and bcl-2, while c-erbB2 overexpression and c-myc amplification was found in both cyclin D1(high) and E(high) tumours. Using transfected cell lines overexpressing cyclin E, cyclin E(high) and D1(high) tumours were mimicked and the cyclin D1(high) cell line normalized the cyclin E kinase activity by an induction and redirection of p21 and p27 to the cyclin E complex whereas cyclin E(high) cell lines obtained increased kinase activity without redirection of inhibitors. Based on differences in genetic aberrations as well as function of the pRb node we therefore propose a model in which cyclin D1(high) and cyclin E(high) tumours represent two alternative mechanisms to inactivate the pRb pathway and thereby achieve unrestrained growth in the tumorogenesis of breast cancer. PMID- 12096345 TI - Nuclear localization is required for induction of apoptotic cell death by the Rb associated p84N5 death domain protein. AB - The mechanisms utilized to transduce apoptotic signals that originate from within the nucleus, in response to DNA damage for example, are not well understood. Identifying these mechanisms is important for predicting how tumor cells will respond to genotoxic radiation or chemotherapy. The Rb tumor suppressor protein can inhibit apoptosis triggered by DNA damage, but how it does so is unclear. We have previously characterized a death domain protein, p84N5, that specifically associates with an amino-terminal domain of Rb protein. The p84N5 death domain is required for its ability to trigger apoptotic cell death. Association with Rb protein inhibits p84N5-induced apoptosis suggesting that it may be a mediator of Rb's effects on apoptosis. Unlike other death domain-containing apoptotic signaling proteins, however, p84N5 is localized predominantly within the nucleus of interphase cells. Here we test whether p84N5 requires nuclear localization in order to trigger apoptosis. We identify the p84N5 nuclear localization signal and demonstrate that nuclear localization is required for p84N5-induced apoptosis. To our knowledge, this identifies p84N5 as the first death-domain containing apoptotic signaling protein that functions within the nucleus. By analogy to other death domain containing proteins, p84N5 may play some role in apoptotic signaling within the nucleus. Further, p84N5 is a potential mediator of Rb protein's effects on DNA damage induced apoptosis. PMID- 12096346 TI - Carcinogen-induced pancreatic lesions in the mouse: effect of Smad4 and Apc genotypes. AB - Mutations in the tumour suppressor genes SMAD4 (DPC4, deleted in pancreatic cancer locus 4) and adenomatous polyposis coli (APC) have been implicated in the development of pancreatic cancer in humans. Treatment of wild-type, Smad4(+/-), Apc(Min/+) or Apc(Min/+)Smad4(+/-) mice with N-Nitroso-N-Methyl Urea (NMU) results in abnormal foci in pancreatic acinar cells characterized by increased levels of beta-catenin. Previously such foci have been shown to be the precursors of pancreatic neoplasia. Interestingly, only NMU-treated Apc(Min/+)Smad4(+/-) mice exhibit a significant increase in abnormal pancreas, which was found to be due to increased number of abnormal foci rather than increased focus size. A range of foci sizes were analysed, but only smaller abnormal foci were characterized by morphological nuclear atypia. These studies suggest functional co-operation between TGF-beta and Wnt signalling pathways in the suppression of pancreatic tumorigenesis in the mouse. PMID- 12096347 TI - Frequent mutations of Fas gene in nasal NK/T cell lymphoma. AB - Fas (Apo-1/CD95) is a cell-surface receptor involved in cell death signaling through binding of Fas ligand. Mutation of Fas gene in lymphoid cells results in accumulation of these cells, which might thus contribute to lymphomagenesis. We examined the open reading frame of Fas cDNA in 14 cases of nasal NK/T-cell lymphoma. Mutations of Fas gene were detected in seven (50%) of 14 cases which comprised four frameshift, two missense, and one silent mutations. Frameshift mutations were caused by insertion of 1 bp (A) at nucleotide 1095 in two cases and by deletion of 1 bp at nucleotide 597 and at 704, respectively, in one each. Mouse T-cell lymphoma cells transfected with two missense mutated genes and frameshift mutations caused by insertion of 1 bp (A) at nucleotide 1095 were resistant to apoptosis induced by the anti-Fas antibody. These findings suggested that accumulation of lymphoid cells with Fas mutations provides a basis for the development of nasal NK/T-cell lymphoma. PMID- 12096348 TI - MLL-SEPTIN6 fusion recurs in novel translocation of chromosomes 3, X, and 11 in infant acute myelomonocytic leukaemia and in t(X;11) in infant acute myeloid leukaemia, and MLL genomic breakpoint in complex MLL-SEPTIN6 rearrangement is a DNA topoisomerase II cleavage site. AB - We examined the MLL translocation in two cases of infant AML with X chromosome disruption. The G-banded karyotype in the first case suggested t(X;3)(q22;p21)ins(X;11)(q22;q13q25). Southern blot analysis showed one MLL rearrangement. Panhandle PCR approaches were used to identify the MLL fusion transcript and MLL genomic breakpoint junction. SEPTIN6 from chromosome band Xq24 was the partner gene of MLL. MLL exon 7 was joined in-frame to SEPTIN6 exon 2 in the fusion transcript. The MLL genomic breakpoint was in intron 7; the SEPTIN6 genomic breakpoint was in intron 1. Spectral karyotyping revealed a complex rearrangement disrupting band 11q23. FISH with a probe for MLL confirmed MLL involvement and showed that the MLL-SEPTIN6 junction was on the der(X). The MLL genomic breakpoint was a functional DNA topoisomerase II cleavage site in an in vitro assay. In the second case, the karyotype revealed t(X;11)(q22;q23). Southern blot analysis showed two MLL rearrangements. cDNA panhandle PCR detected a transcript fusing MLL exon 8 in-frame to SEPTIN6 exon 2. MLL and SEPTIN6 are vulnerable to damage to form recurrent translocations in infant AML. Identification of SEPTIN6 and the SEPTIN family members hCDCrel and MSF as partner genes of MLL suggests a common pathway to leukaemogenesis. PMID- 12096349 TI - Testing for population subdivision and association in four case-control studies. AB - Population structure has been presumed to cause many of the unreplicated disease marker associations reported in the literature, yet few actual case-control studies have been evaluated for the presence of structure. Here, we examine four moderate case-control samples, comprising 3,472 individuals, to determine if detectable population subdivision is present. The four population samples include: 500 U.S. whites and 236 African Americans with hypertension; and 500 U.S. whites and 500 Polish whites with type 2 diabetes, all with matched control subjects. Both diabetes populations were typed for the PPARg Pro12Ala polymorphism, to replicate this well-supported association (Altshuler et al. 2000). In each of the four samples, we tested for structure, using the sum of the case-control allele frequency chi(2) statistics for 9 STR and 35 SNP markers (Pritchard and Rosenberg 1999). We found weak evidence for population structure in the African American sample only, but further refinement of the sample, to include only individuals with U.S.-born parents and grandparents, eliminated the stratification. Our examples provide insight into the factors affecting the replication of association studies and suggest that carefully matched, moderate sized case-control samples in cosmopolitan U.S. and European populations are unlikely to contain levels of structure that would result in significantly inflated numbers of false-positive associations. We explore the role that extreme differences in power among studies, due to sample size and risk-allele frequency differences, may play in the replication problem. PMID- 12096351 TI - Quality of life issues in patients with essential thrombocythemia and polycythemia vera. AB - Essential thrombocythemia and polycythemia vera are both chronic progressive myeloproliferative disorders of insidious onset. If the excessive production of red cells and/or platelets is controlled, patients with these disorders may have prolonged survival. However, the clinical course of these patients can be complicated by a variety of events, including thrombotic episodes, bleeding episodes, arthropathies, pruritus, weakness, weight loss, neurologic impairment, erythromelalgia, fever, abdominal pain, and the life-threatening consequences of progression to myelofibrosis and/or acute leukemia. Effective control of hematopoiesis by phlebotomy or a variety of therapeutic agents has resulted in a reduction or elimination of many of these clinical events, but has not altered the evolution to myelofibrosis or acute leukemia. Use of each of these therapeutic strategies is also associated with a range of adverse events. Monitoring overall survival or a reduction in the frequency of clinical events has previously served as a means of assessing the results of these therapeutic interventions. Quality-of-life instruments have not been applied in a systematic fashion to the evaluation of outcomes in patients with these chronic myeloproliferative disorders. Quality-of-life assessments evaluate not only the state of well-being of a patient that results from an assessment of the individual's ability to perform everyday activities, which are reflective of physical, psychological, and social well-being, but also patient satisfaction with the control of disease and/or treatment-related symptoms. Quality-of-life instruments have been used to assess the clinical course of patients suffering from a variety of disorders, ranging from cancer to renal failure to chronic fatigue syndrome. Information about quality-of-life outcomes can contribute to the evaluation of variations in dose and timing of administration of therapeutic agents. It is possible that the side effects of a particular therapy may outweigh the disease regression achieved with a particular therapy. In the future, quality of-life instruments may prove useful in prospectively evaluating therapeutic end points in patients with essential thrombocythemia and polycythemia vera. PMID- 12096353 TI - The risk of thrombosis in essential thrombocythemia and polycythemia vera. AB - Thrombotic events are present in 20% to 50% of patients with polycythemia vera (PV) and essential thrombocythemia (ET) at diagnosis and involve major vessels and the microcirculation. Establishing the precise risk of thrombosis in ET and PV is difficult. However, estimates may be obtained from retrospective and prospective studies, and some risk factors have been defined. In ET, increasing age and previous thrombosis emerge as major factors. The level of the platelet count per se does not correlate with thrombotic incidence. However, there is substantial evidence that adequate control of the platelet count reduces the frequency of thrombosis. While aspirin is usually effective at relieving vasomotor and microvascular occlusive symptoms or signs, there is only limited evidence that its use reduces the risk of larger vessel thrombosis. Other suggested risk factors include apparent clonal hematopoiesis in younger patients, hypercholesterolemia, and cigarette smoking. In PV, the overall impression is that the incidence of thrombosis is greater than in ET. As in ET, increasing age and previous thrombosis are significant risk factors. In addition, adequate reduction of the hematocrit reduces the incidence of thrombosis. Identifying the risk associated with thrombocytosis in PV is confounded by the overall myeloproliferation usually found in these patients and the fact that the thrombocytosis is not usually as marked as in ET. However, by analogy with ET, one could argue that the risk associated with the thrombocytosis in ET is equally applicable to PV. In support of this, there is overwhelming evidence that patients treated with adequate myelosuppression with complete normalization of their blood counts have the lowest thrombotic incidence. Essential thrombocythemia and PV patients presenting with thrombosis should be investigated for other congenital or acquired prothrombotic conditions. Establishment of the presence of one or the other of these may alter the long-term management of these patients with the use of either aspirin or anticoagulation in addition to cytoreduction. Thrombotic risk has emerged as a major factor in stratifying patients with myeloproliferative disease. While some clear thrombotic risk factors have been defined, there is a need to identify further additive risk factors in patients considered to be at low risk of thrombosis. PMID- 12096352 TI - Therapeutic options for essential thrombocythemia and polycythemia vera. AB - Several options exist for treating essential thrombocythemia and polycythemia vera. One approach is to assign the patient to a risk category from which treatment recommendations follow. The principal risks of essential thrombocythemia include thrombosis, major hemorrhage, and conversion to leukemia or myelofibrosis. Risk factors for thrombosis include age and prior thrombosis. Smoking and obesity have been implicated in isolated series. High-risk patients with essential thrombocythemia can be defined as those 60 years of age or older or those who have had a thrombosis at any age. These patients should be treated with hydroxyurea. If hydroxyurea cannot be tolerated, anagrelide and interferon alpha (IFN-alpha) are alternatives. Low-dose aspirin (40 to 325 mg) can be used for patients whose platelet counts are < 1,500 x10(9)/L. Low-risk patients are those less than 60 years old who have not had thrombosis, who have no cardiovascular risk factors, and whose platelet counts are < 1,500 x 10(9)/L. These patients can be observed or placed on low-dose aspirin. Intermediate-risk patients are those less than 60 years who have not had thromboses, but who have platelet counts > 1,500 x 10(9)/L or who have significant cardiovascular risk factors. These patients should have their risk factors treated and may be given low-dose aspirin if the platelet count is < 1,500 x 10(9)/L. They can be observed or treated with anagrelide, hydroxyurea, or IFN-alpha. The Mayo Clinic experience suggests that no specific treatment affects outcomes of pregnancies. In high-risk pregnant women who need treatment, IFN-alpha is used. The principal risks of polycythemia vera are thrombosis, postpolycythemia myeloid metaplasia, and acute leukemia. Risk factors for thrombosis include age, the use of phlebotomies, the rate of phlebotomies, and a prior history of thrombosis. Platelet counts have not been definitively linked to an increased risk of thrombosis. High-risk polycythemia vera patients are those 60 years of age or older (some groups use 70 years) or those of any age who have had thrombosis. They should be treated with phlebotomy and hydroxyurea or IFN-alpha. Selected patients may be treated with anagrelide. A typical target range for phlebotomy is a hematocrit of < 42% for women and < 45% for men. Low-dose aspirin can be used if the platelet count is < 1,500 x 10(9)/L. Low-risk patients are those less than 60 years old who have had no thrombosis, no cardiovascular risk factors, and whose platelets are < 1,500 x 10(9)/L. These patients can be managed with phlebotomy alone or phlebotomy and low-dose aspirin. Intermediate-risk patients are those who are less than 60 years old, who have not had thrombosis, but who have platelet counts > 1,500 x 10(9)/L or who have cardiovascular risk factors. The cardiovascular risk factors should be treated, along with phlebotomy alone or with IFN-alpha. Low-dose aspirin is reasonable for those with platelet counts < 1,500 x 10(9)/mL. Anagrelide can be used with phlebotomy in selected patients. Women of childbearing age who are in the low-risk or intermediate-risk group can be treated with phlebotomy alone and low-dose aspirin if the platelet count is < 1,500 x 10(9)/L. For high-risk patients or pregnant patients, IFN-alpha can be added. PMID- 12096354 TI - Other secondary sequelae of treatments for myeloproliferative disorders. AB - Polycythemia vera (PV) and essential thrombocythemia (ET) are chronic disorders for which there are no medical cures. Clinical sequelae of PV and ET fall into three categories: primary, such as thrombosis and hemorrhage; secondary, resulting from disease progression or treatment. The decision whether to treat the patient is based on the sequelae of no treatment versus short- and long-term toxicities of the three classes of drugs available for treatment: hydroxyurea, interferon-alpha, and anagrelide. Thrombosis is the most common short-term sequelae of untreated disease; the risk increases with age and after the first thrombotic complication. Hydroxyurea, a nonalkylating myelosuppressive agent, is mutagenic and probably leukemogenic over 5 to 15 years, which makes it unsuitable for treating most younger patients. Interferon-alpha, a cytokine that is myelosuppressive and immunomodulatory, has been shown to have a therapeutic effect in both PV and ET. Tolerance to the initial flu-like symptoms of interferon-alpha is usually developed, but dose-limiting symptoms of anorexia, asthenia, and neuropsychiatric disease can occur, along with exacerbation or development of autoimmune diseases. Anagrelide, a quinazoline that inhibits cyclic nucleotide phosphodiesterase, inhibits platelet aggregation and has an idiosyncratic effect of inhibiting megakaryocyte maturation and platelet budding at doses below those that affect platelet function. This agent is a vasodilator with positive inotropic activity and a side-effect profile that may include palpitations, forceful heartbeat, tachycardia, and headache. One in four patients develop fluid retention and/or edema that are controllable with diuretic therapy. Dizziness is frequent, but mild. Because these side effects usually abate in 2 to 4 weeks, successful management of patients taking anagrelide depends on encouraging them to maintain therapy. The availability of these three classes of drugs with differing modes of action suggests that combination therapy may offer the opportunity to achieve better control of proliferation while reducing short term side effects as well as the risks of dose-related cumulative sequelae. PMID- 12096355 TI - Cost-effectiveness considerations in the treatment of essential thrombocythemia. AB - Factors that influence the choice of anagrelide, hydroxyurea, or interferon-alfa (IFN-alpha) for treatment of essential thrombocythemia include efficacy, toxicity, and cost. Anagrelide has the US Food and Drug Administration's approval to be used for treating patients with thrombocythemia secondary to chronic myeloproliferative disorders. In contrast, the use of IFN-alpha and hydroxyurea are considered "off-label." We performed an incremental cost-effectiveness analysis to compare anagrelide, hydroxyurea, and IFN-alpha for treating essential thrombocythemia, in terms of estimated impact on life expectancy. The case used for this analysis was of a 40-year-old man with essential thrombocythemia. Clinical assumptions were based on information obtained from nonrandomized clinical trials, and the economic assumptions were derived from information abstracted from observational studies. Lifelong treatment use of anagrelide versus hydroxyurea would cost approximately $72,000 per additional year of life gained, while the use of IFN-alpha was found to be both more costly and less effective than anagrelide. The results were very sensitive to the risk of leukemia caused by hydroxyurea, with an incremental cost-effectiveness of anagrelide compared with hydroxyurea of $156,969 per additional year of life gained if the lifetime leukemia risk drops from a baseline of .08 to.05. Given that many commonly used medical interventions cost in the range of $50,000 to $100,000 per year of life gained, and the generally poor outcome associated with treatment-related leukemia that can result from hydroxyurea, anagrelide could be considered a therapeutic alternative that is clinically effective at an acceptable cost. PMID- 12096356 TI - Global nursing language: making international nursing visible. PMID- 12096357 TI - Risks to students' civil liberties. PMID- 12096358 TI - The changing face of nursing. PMID- 12096359 TI - Differentiation: practice versus services. PMID- 12096360 TI - Genomic-based health care in nursing: a bidirectional approach to bringing genetics into nursing's body of knowledge. AB - Nurses can expect to become partners with patients and their families as genomic based health care brings decisions involving gene-based diagnostics and therapeutics into commonplace practice. All health professionals need genetic literacy to continue competence in clinical practice in the future. This article explores the efforts nursing has undertaken during the past decade to integrate human and clinical genetics into practice and scholarship. Genetics nursing education literature in the second half of the 20th century was analyzed. A focused survey of 15 genetics nurses and nursing leaders regarding key national initiatives, genetics research training programs, and genetics education models was conducted. Nursing has taken a bidirectional approach (through grass roots and top-down initiatives) to implementing change and advancing genetics in nursing practice, research, and education. For optimal leadership in genetics education for nursing and other disciplines, bidirectional efforts must continue and concentrate on bringing genetics into clinical practice and scholarship as genomic-based health care spreads worldwide. Evidence of nursing's interdisciplinary leadership is recognized. Recommendations and strategies for continued nursing leadership and programs that build on previous work are presented. PMID- 12096361 TI - Strengthening end-of-life care through specialty nursing certification. AB - The purpose of this study was to determine the adequacy of content related to end of-life (EOL) care in materials used in the nursing certification process across clinical nursing specialties. Thirty-eight certification examination blueprints, 18 specialty nursing scope and standards of practice documents, and 28 specialty nursing core curriculum text books were analyzed by using descriptive statistics to determine the quantity and quality of content related to nine critical areas of EOL content contained in them. Fifteen (38 per cent) of the certification examination blueprints contained at least one of the critical EOL content areas. Eight (44 per cent) of the scope and standards of practice documents contained at least one sentence on EOL care. Seven (25 per cent) of the 28 textbooks contained at least one chapter dedicated to EOL care content, and 129.5 (0.8 per cent) of the 15,706 textbook pages reviewed were dedicated to EOL care content. Expert ratings regarding the overall accuracy, currency, and comprehensiveness of EOL content found in the textbooks were poor to good. An increased focus on EOL care in the nursing specialty certification process is warranted. The content of nursing specialty certification examinations has a direct influence on nursing education as well as a significant impact on nursing practice in clinical specialty areas. PMID- 12096362 TI - Strategies for building faculty research programs in institutions that are not research intensive. AB - Colleges and universities without the resources of research-intensive universities face a special challenge to support faculty research. If doctorally prepared faculty are to assume. leadership roles in developing nursing science, deans must be responsive to faculty members' individual and collective responsibility to be active researchers. This article describes efforts on the part of two nursing programs, one in a private and one in a public university, to create an environment that nurtures scholarship of nursing faculty members at these institutions. PMID- 12096363 TI - Assessment of AACN baccalaureate nursing education curriculum model in the People's Republic of China: a transcultural exploratory study. AB - This study examines the relevance of The Essentials of Baccalaureate Education for Professional Nursing Practice (American Association of Colleges of Nursing, 1998) to baccalaureate nursing curriculum development in the People's Republic of China. A Likert-type survey was developed to measure Chinese nurse educators' perceptions regarding the 21 key concepts extracted from The Essentials document in both the present curriculum and an ideal curriculum across three dimensions: importance, cultural relevance, and extent of exposure. Surveys were sent to all known baccalaureate nursing programs in China (N = 22) with a 50 percent return rate. Descriptive statistical analyses supported the universality of the majority of the 21 key concepts from The Essentials document. Some of the concepts, however, were not as readily transferable because of differences in the Chinese sociopolitical and cultural contexts for baccalaureate nursing education. Findings from this study lend support for the potential global relevance of The Essentials document. Additional transcultural and comparative studies are recommended to further test the relevance of The Essentials document in other cultures. Meanwhile, international nurse educators need to be aware of the possible limits of adopting The Essentials document without values clarification, critical discrimination, and adaptation to their home cultures. PMID- 12096364 TI - Effects of program design on the professional socialization of RN-BSN students. AB - The purpose of this study was to examine effects of program design on the bureaucratic and professional role conceptions of registered nurses in baccalaureate nursing programs. Attention to how programs are set up and function has relevance for theory and practice and enables improving frameworks for program planning. The study design embodied qualitative and quantitative elements and used six processes of socialization in organizations as a conceptual guide. Qualitative data collection concerned comparative case studies of three baccalaureate nursing programs and data analysis included narratives detailing program organization and describing student experience. Quantitative matter involved mailed questionnaires to determine graduates' bureaucratic, professional, and service role conceptions. Data analysis (analysis of variance) revealed statistically significant differences in role conceptions (F[3.56] = 4.51) of registered nurses graduating from different baccalaureate programs in nursing. Indications are that nursing programs can be set up to produce equal education and reach certain stated outcomes, yet implicit program processes can impact student learning. PMID- 12096365 TI - Institutional collaboration not competition: preparing family nurse practitioners to serve rural areas. AB - In response to the need for an increased number of family nurse practitioners in rural Texas, The University of Texas at Tyler, Texas Tech University Health Sciences Center, and Midwestern State University entered into a collaborative arrangement to provide advanced education to nurses in outlying areas of the state. The catchment area for the project encompassed 72 counties, representing half the land mass in Texas. This article presents the development of the collaborative model, strategies used for implementation, and evaluation of this collaborative effort. The collaborative family nurse practitioner program is based on collegiality rather than competition to better meet the needs of students and communities in a cost-effective manner. Communication between the institutions and faculty has been a key factor in the success of the program. The addition of nurse practitioners from this project has the potential to greatly increase access to health care in large, underserved populations. PMID- 12096367 TI - Preceptor questioning and student critical thinking. AB - Questioning is fundamental to student learning. Not only does it enable students to elevate their level of thinking, but in the process it also affords them the opportunity to deal with their world intelligently. The practice setting is an environment rich in opportunity for enabling critical thinking through the use of questioning. In the preceptorship experience, preceptors are in a prime position to use questioning behaviors that can challenge the way preceptees think, encourage them to justify or clarify their assertions, promote the generation of original ideas, explanations, or solutions to patient problems, provide mental and emotional tools to help resolve dilemmas, promote discussion, and evaluate learning. This article discusses the importance of preceptor questioning for the development and promotion of student critical thinking. Contextually, the authors draw on the findings of a recent study in which preceptor questioning of the knowledge base, decision making, and actions of the preceptee were found to directly bring about or trigger their critical thinking. This article allows for some further reflection on that process and its contribution to the enhancement of the preceptorship experience. PMID- 12096366 TI - Gender-related influences in nursing education. AB - Issues around gender identity, self-concept, professional education, and women's learning preferences shape the socialization of nurses and nursing education. How gender issues influence learners and nursing education in relation to nursing education for registered nurses are explored and recommendations are made for nursing educators in two broad categories: self-awareness and helping learners learn. The exploration leads to three conclusions: first, it is timely for nurses to continue such explorations for their own practice, and to share how to help other nurses cope with visible and invisible gender-related issues; second, nurses can share strategies to address the influence of gender issues in nursing education, and, third, the nursing profession benefits from drawing on available resources to discuss gender issues. Although reflections on these issues have been shaped by working with registered nurse learners, they are equally relevant to basic nursing students and practicing nurses. PMID- 12096369 TI - Editorial: safety and critical care. PMID- 12096370 TI - Building safety into ICU care. AB - The Institute of Medicine's (IOMs) report, "To Err is Human," recently addressed patient safety in the United States, alerting the nation to the need for improved systems of health care. Seven main findings were addressed in this report, we focus on 3: (1) patient safety is a nationwide problem, (2) health care workers are not to blame, and (3) safety and harm are products of care systems. This article discusses systems in intensive care units (ICUs) and how these systems affect patient safety. We use a case example to outline the complex chain of medical and administrative system failures that can result in an adverse event. Then we discuss evidence linking ICU organizational characteristics with patient safety, focusing on how safer systems in ICUs can directly improve patient care. PMID- 12096371 TI - ICU incident reporting systems. AB - Intensive care is one of the largest and most expensive components of American health care. Studies suggest that errors and resulting adverse events are common in intensive care units (ICUs). The incidence may be as high as 2 errors per patient per day; 1 in 5 ICU patients may sustain a serious adverse event, and virtually all are exposed to serious risk for harm. Theories of error developed in aviation and other high-risk industries suggest that errors are likely to occur in all complex systems. Reporting of incidents, including both adverse events and near misses, is an essential component for improving safety. Voluntary, confidential reporting is likely to be more important than mandatory reporting. There have been a few efforts to apply such systems in medicine. In intensive care, the Australian Incident Monitoring System (AIMS)-ICU has been the most prominent. We have designed a Web-based ICU Safety Reporting System (ICUSRS). The goal is to identify high-risk situations and working conditions, to help change systems, and reduce the risk for error. The analysis and feedback of reports will inform the design of interventions to improve patient safety. The effort is aided substantially by collaboration with the 30 participating ICUs and important stakeholders including the Society of Critical Care Medicine, the American Society for Health-care Risk Management, the Food and Drug Administration Center for Devices and Radiological Health, the Foundation for Accountability, and the Leapfrog Group. A demonstration and evaluation of the system is underway, funded by the Agency for Healthcare Re-search and Quality. PMID- 12096372 TI - Venous thromboembolism and its prevention in critical care. AB - BACKGROUND: Evidence-based guidelines for the prevention of venous thromboembolism (VTE) are available for most major surgical and medical patient groups. Such guidelines have not been established for critically ill patients. OBJECTIVE: To perform a systematic review of the prevalence of deep vein thrombosis (DVT), the efficacy of thromboprophylaxis, and the rates of thromboprophylaxis use in critically ill patients. METHODS: Computerized literature search for relevant studies meeting prespecified criteria. RESULTS: The rates of objectively confirmed DVT in 4 prospective studies ranged from 13% to 31%. We identified only 3 randomized trials (1 in abstract form) of thromboprophylaxis in critical care unit patients. These studies show the efficacy of low-dose heparin and low molecular weight heparin compared with no prophylaxis; however, we found no trials comparing these 2 interventions. Eleven compliance studies reported that some form of thromboprophylaxis was used in 33% to 100% of critically ill patients, although only 1 study addressed the issue of appropriate prophylaxis use. CONCLUSIONS: Data on the epidemiology of VTE and its prevention in critically ill patients are very limited. Further research is needed to better define patient risk factors for VTE, optimal methods of thromboprophylaxis, and strategies to improve compliance with prophylaxis recommendations. In the meantime, prevention strategies, shown to be effective in other related patient groups, and general principles of individual pharmacotherapy should guide the routine use of prophylaxis during critical illness. PMID- 12096373 TI - Perioperative beta-blockers in high-risk patients. AB - One major risk to patients in the preoperative period is that of myocardial ischemia or infarction and cardiovascular death in high-risk patients. Historically, attempts to decrease the incidence of perioperative cardiac complications have focused on preoperative evaluation and identification of patients at risk for complications with referral for additional testing and/or revascularization. Evidence suggests that the use of perioperative beta-blockers in high-risk individuals can reduce the incidence of perioperative cardiac events. The Agency for Healthcare Research and Quality has identified that the use of perioperative beta-blockers can reduce perioperative morbidity and mortality. The focus of this article is to describe the evidence supporting perioperative beta-blocker use, to discuss potential barriers to their use, and to propose a strategy to improve their use. PMID- 12096374 TI - Applying the science to the prevention of catheter-related infections. AB - BACKGROUND: Catheter-related bloodstream infection (CR-BSI) remains a leading cause of nososcomial infection, despite the fact that many CR-BSIs are preventable. Simple principles of infection control and the use of novel devices to reduce these infections are not uniformly implemented. OBJECTIVE: To review and summarize the evidence that promotes strategies to prevent CR-BSIs. DATA SOURCES: The Medline database and bibliographies of review articles and book chapters were searched for relevant articles. STUDIES INCLUDED: In vitro and in vivo laboratory-based studies, controlled clinical trials, prospective interventional trials, and epidemiologic investigations. OUTCOME MEASURES: Reduction in CR-BSI, catheter colonization, tunnel infection, or local site infection. SYNTHESIS: The recommended prevention strategies with strong supportive evidence include educating and training of health care providers who insert and maintain catheters; using full barrier precautions during central venous catheter insertion; using a 2% chlorhexidine preparation for skin antisepsis; eliminating the practice of scheduled replacement of central venous catheters for prevention of infection; and using antiseptic/antibiotic impregnated short-term central venous catheters. CONCLUSION: Simple interventions can reduce the risk for serious catheter-related infection. Health care provider awareness and adherence to these prevention strategies is critical to reducing the risk for CR-BSI, improving patient safety, and promoting quality health care. PMID- 12096375 TI - Pressure relief bedding to prevent pressure ulcer development in critical care. AB - One major risk for the critically ill patient is the development of pressure ulcers during the intensive care unit (ICU) stay. These patients have many of the risk factors for the development of pressure ulcers including reduced mobility/activity, medications, neurologic deficits, increasing age, incontinence, decreased mental status, poor nutrition, pressure, shear forces, and friction. Pressure ulcers are known to be costly for the health care system and delay recovery in many patients. Different strategies have been advocated for the prevention of pressure ulcers, and the Agency for Healthcare Research and Quality (AHRQ) has identified the use of pressure relief bedding as a means to prevent the development of pressure ulcers during hospitalization. The use of pressure relief bedding has received the most research attention to date. The focus of this article is to describe the state of the current research in this area and how this applies to critical care. Development of protocols and guidelines for the use of pressure ulcer preventing strategies are important to improve the quality of care in the ICU. There is still a need to examine the impact of the evidence of pressure ulcer prevention in the ICU and this review should help to build a framework for future research and protocol development. PMID- 12096376 TI - Use of ultrasound to place central lines. AB - CONTEXT: Placement of central venous catheters (CVCs) is an integral part of care for the critically ill patient but is associated with significant morbidity when using the traditional landmark method. The use of real-time ultrasound to guide line placement has been developed in hopes of avoiding this morbidity. OBJECTIVE: The objectives of this article are 2-fold. The first is to determine the relative effectiveness of the use of real-time ultrasound to place CVCs compared with the use of landmarks alone. The second is to discuss the merits of future study to increase the use of this technology. DATA SOURCES: Medline from 1966 to 2001, personal files, 2 prior systematic reviews, and reference lists of selected articles. STUDY SELECTION: Studies were included if: (1) study design was a controlled trial, (2) patients required placement of a CVC, (3) the interventions were real-time ultrasound versus standard landmark-guided line placement, and (4) outcomes included at least 1 of failure to place catheter, success of first attempt, number of attempts, time to catheter placement, or complication rate. DATA SYNTHESIS: Eighteen trials were identified. Pooled results showed a significant reduction in failure rate (risk difference, -.12, 95% confidence interval [CI], -.18 to -.06), number of attempts (risk reduction, 1.41, 95% CI, 1.15-1.67), and arterial puncture rate (risk difference, -.07, 95% CI, -.10 to .03). The number of successful venous cannulations on first attempt were higher using ultrasound (risk difference,.24, 95% CI,.08-.39). No difference was found in time to insertion. Significant heterogeneity of study results was found for most analyses. Subgroup analyses suggested that ultrasound improved outcomes most convincingly using external probes, for internal jugular vein cannulation, and when used by clinicians less experienced at line placement. CONCLUSIONS: Adoption of real-time ultrasound to guide CVC placement has the potential to improve successful line placement and minimized complications. It can improve patient safety. However, there are significant cost concerns and the reported adverse events are generally minor and easy to treat. Before creating study protocols to increase usage of this technology, both current usage and cost effectiveness should be determined. PMID- 12096378 TI - What patients want ... are educated dentists. PMID- 12096377 TI - Changing physician behavior: a review of patient safety in critical care medicine. AB - The publication of the Agency for Healthcare Research and Quality (AHRQ) report in July 2001 entitled "Making Health Care Safer: A Critical Analysis of Patient Safety Practices," represents a significant perceptual change in health care ideology. It can be argued that this compilation recognizes not only that medical errors occur in the health care system, but also that there are significant learning opportunities that may arise in the identification of these errors that are otherwise known as medical misadventures. The report concluded and outlined a series of 11 highly rated practices whose usage are associated with increased safety. The AHRQ report also articulated that there is a need to investigate methods used to align medical practice with evidence regarding patient safety. In other words, after the identification of the 11 priority safety practices, it is thus important to determine the most effective methods to change physician behavior toward these practices that will intuitively result in increased safety performance. Five different educational-based strategies have been identified as techniques to change physician behavior: (1) Academic Detailing, (2) Audit and Feedback, (3) Local Opinion Leaders, (4) Reminder Systems, and (5) Printed Material. This article reviews these strategies in the context of critical care medicine and offers some opinions regarding setting the future research agenda in this investigative field. PMID- 12096379 TI - Restorative management of the worn dentition: 2. Localized anterior toothwear. AB - This is the second paper in a four-part series detailing the relative merits of the treatment strategies, clinical techniques and dental materials for the restoration of health, function and aesthetics for the dentition. In this paper the management of wear in the anterior dentition is discussed, using three case studies as illustration. PMID- 12096380 TI - Iron and vitamin B12 deficiency anaemia in a vegetarian: a diagnostic approach by enzyme-linked immunosorbent assay and radioimmunoassay. AB - This article presents the case of a 46-year-old vegetarian who had a painful dry socket in the left third molar areas. Since the patient's general appraisal was anaemic, investigations for haematological status, folic acid and vitamin B12 were performed. The results revealed that the patient was severely iron deficient and slightly vitamin B12 deficient. PMID- 12096381 TI - Preserving the vital pulp in operative dentistry: 4. Factors influencing successful pulp capping. AB - The sequence of factors that mediate pulp inflammation and necrosis are unclear, and controversy surrounds the effects of different pulp capping materials on exposed pulps. Clinicians have few quantitative studies that rank the in vivo pulp capping effects of commonly used restorative materials. PMID- 12096384 TI - Assessing restorative dental materials: 2. Questions for a dental sales representative. AB - Many methodologies are used during the testing of dental materials, although different tests are suitable for different materials and clinical situations. This paper discusses factors relevant to the choice of a dental material, and suggests questions that a practitioner would find helpful to ask a dental sales representative, such as ease of use, effect on the environment and the relevance of fluoride release. PMID- 12096383 TI - Preference for venepuncture site: should the patient be consulted? AB - A questionnaire-based study was undertaken to ascertain if patient preferences were being met in clinical practice by investigating patients' past experience of venepuncture and patient and operator preference for venepuncture site. Patients and operators expressed a preference for either the dorsum of the hand or the antecubital fossa as sites of venous access. Patients who expressed a preference for the dorsum of the hand were more likely to have received venepuncture in their chosen site, although the likelihood of venepuncture failure is higher in the dorsum of the hand than in the antecubital fossa. PMID- 12096382 TI - Late lower arch crowding: the aetiology reviewed. AB - This article reviews the causes of the crowding that commonly occurs, particularly in the lower arch, after eruption of the second permanent molars. Factors discussed include mesially directed forces, in treated and untreated subjects, distally directed forces, occlusal changes, direction of eruption, tooth morphology, periodontal forces, and degenerative connective tissue changes. PMID- 12096385 TI - The use of teledentistry to provide GDPs with advice in orthodontics. AB - This article describes a pilot trial to determine whether the transmission of digitized clinical records of potential orthodontic cases supported by access to video and data conferencing may help to reduce the current high level of inappropriate referrals to consultant orthodontists in the UK. PMID- 12096388 TI - Patient confidentiality and privacy in cyberspace: the nurse's obligation. PMID- 12096387 TI - Insurance protection for licensure disciplinary actions. PMID- 12096389 TI - [Treatment of iron deficiency in preschool children with a weekly dose of ferrous sulfate]. AB - INTRODUCTION: Iron deficiency is the most frequent haematological disease during the paediatric age and is a health public problem; the iron deficiency has a special interest during early development of human beings; in recent years the group of research recommend intermittent treatment, with ferrous sulphate every week because is more easily adhesion to treatment. OBJECTIVE: To know iron concentration in plasma after treatment, given oral ferrous sulphate daily and single dose every week. METHODS AND MATERIAL: We established two groups A and B, with ten iron deficiency pre-school children each one. RESULTS: Group A: the children received oral ferrous sulphate twice a day during three months; the doses were 3.5 mg of ferric iron per kilogram of weight in children with ages less of three years old, and 2.5 mg/kg/day in children with three to five years old. After the treatment all the patients improved haemoglobin, ferritin an iron concentrations (p = 0.001). Group B: The patient received the some dose per kilogram of weight like that of group A, in a single dose weekly for three months, there was an improvement in haemoglobin not reaching normal levels, half of the patients improved iron and ferritin observing statistical significance; haemoglobin (p = 0.001) iron (p = 0.062) and ferritin (p = 0.01). CONCLUSION: The administration of ferrous sulphate daily provide an positive balance of iron status for the group A. Group B: that they received iron weekly dose showed changes to positive balance of iron status with a statistical significance, it is probably that the half doses of ferrous sulphate also improve the anaemia by iron deficiency. PMID- 12096390 TI - [Palliative care in advanced cancer. A 7-year experience at the Dr. Juan I. Menchaca Civil Hospital of Guadalajara, Jalisco, Mexico]. AB - We describe 7 years of experience in terminal cancer care provided by a multi professional palliative care team. A total of 995 persons requested our services during this period. We included 649 patients for whom we had sufficient information for this study. Average age was 58 years; 60.2% patients were female, whereas 39.7% were male. Underlying disease was cervical cancer followed by breast cancer and prostate, etc. The most frequent symptom on admission was pain followed by weakness, loss of weight, anorexia, and emesis (nausea and vomiting). Average stay in the program of patients who died (406) was 67 days with a range of 1-707 days. We compare symptom incidence and model of care with other palliative care and hospice programs in the world. Cost-Benefit and future implementation of similar programs in Mexico is discussed. PMID- 12096391 TI - [Peripheral blood hematopoietic cell transplant using immunosuppressive chemotherapy without bone marrow destruction: "minitransplant"]. AB - Using a nonmyeloablative, immunosuppressive, fludarabine (FLU)-base conditioning regimen, we have performed allogeneic peripheral blood stem cell transplants in 17 patients (six with chronic granulocytic leukemia, four with acute myelogenous leukemia, five with acute lymphoblastic leukemia, one with myelodysplasia and one, with thalassemia major). Conditioning regimen consisted of FLU/busulfan/cyclophosphamide or FLU melphalan. To avoid graft vs. host disease (GVHD), cyclosporine and methotrexate were used. Median granulocyte recovery time to 0.5 x 10(9) was 11 days, whereas median platelet recovery time to 20 x 10(9) was 12 days. Seven patients did not need red blood cell transfusions and four did not need platelet transfusions. In thirteen individuals (76%), the procedure could be completed fully on an outpatient basis. Follow-up times range between 1 and 14 months. Five of 17 patients developed acute GVHD whereas 4/10 developed chronic GVHD. The 14-month survival (SV) is 70% and median SV is not reached. Five patients (29%) have died, three due to relapse of the disease and two due to GVHD. The transplant-related mortality was 5.8%. This procedure is substantially less costly than its counterpart, using in-hospital myeloablative conditioning regimens, and may represent another approach in management of patients requiring allogeneic stem cell transplant. PMID- 12096393 TI - [Functional neuroimaging. A combination of anatomy and physiology]. PMID- 12096392 TI - [Risk factors for therapeutic noncompliance of patients with epilepsies]. AB - Inadequate compliance is a major contributor to unsuccessful treatment in epilepsies. To establish risk factors associated with therapeutic non-compliance in patients with epilepsy, we carried out a case-control study, nested into a cohort, with thirteen factors possibly implicated in therapeutic non-compliance. The patient's general characteristics, the illness, and patient-practitioner relationship were studied. Patients were followed during 6 months; during this time, serum levels and pill counts were registered. Of 150 patients, 66 were non compliers and 84 were compliers. Seven of thirteen factors were statistically different with an odds ratio greater than 3 (p < 0.05). However, after log-lineal regression analysis, only the total number of pills per day and the subject's intellectual level were significant. These two factors increase 3.66 times the risk of non-compliance. We conclude that epileptic patients with a low intellectual level and more than three prescribed pills per day have a 3.66 times greater risk of non-compliance to anti epileptic treatment. PMID- 12096394 TI - [Vaginal cytology: the importance of the transformation zone and how to obtain an adequate sample]. AB - Cytology smear is the test most frequently used to detect cancer of the uterine cervix. Much is written concerning it in its three phases, analytical, post analytical, and preanalytical; the later implies sampling and preservation and is the purpose of this review. According to the national literature, result of the sampling is deficient in 64% of cases. Many instruments have been developed to take good samples, and statistically there is no difference between these in terms of achieving the goal of taking a good sample; despite this in Mexico bad samples persist. In this paper it is analyzed the source of these errors that are principally from ignoring the anatomy of the cervix, the handling and fixation of the sample, among others, and some observations are made that are not described in other articles in the same field. Some of the most popular instruments available in Mexico to take samples are the endocervical brush, spatula, and brush (Pappette) here described; their advantages and disadvantages are included, according to the type of cervix in which is going to be used, stressing some details concerning the transformation zone and how to select the best instrument. The results improved with these observations from 45 to 77% in taking good samples. PMID- 12096395 TI - [A 46-year-old woman with clinical characteristics of growth hormone excess]. PMID- 12096396 TI - [Hepatic hematoma caused by fascioliasis]. AB - The clinical presentation of two patients was carried out, with the history of having consumed in their diet watercress, and presented digestive and general symptoms. One of the patients presented hepatomegalia; but both presented jaundice and eosinophilia; CAT scan was suggestive in both cases of the presence of hepatic hematomas, confirmed by laparoscopic study and surgical intervention. The definitive diagnosis of hepatic hematoma by fasciolasis was obtained during the operation and the postoperative study, added to the other important complementary exams in this entity. PMID- 12096397 TI - [Desmoid fibroma of the mandible in a child and one-stage reconstruction]. AB - A 12-year-old patient case is reported, presented with a desmoid fibroma of the left pterygoid region, extended to the cortical and endosteal regions of the mandible, reconstructed immediately with a free peroneal flap, recovering with this both form and function. To date, 3 years from surgery, he has no complications. Evolution of this case and a brief review of the literature is reported. It is concluded that free peroneal graft for mandible reconstruction is safe and has excellent results, both aestethic and functional. PMID- 12096398 TI - [Notes on ancient Islamic medicine]. AB - Arab medicine arose as a consequence of the assimilation and breeding of Hellenistic medicine, particularly of Galenic medicine. It reached its high point between the X and XII centuries and, after the XIII century, lost all creative capabilities. Nevertheless, it achieved the status of being an incentive for European medieval medicine. Some aspects of the medical teaching and publications of the most distinguished Moslem physicians, such as Rhazes (865-932), Avicenna (980-1037), and Averroes (1126-1198) are described. The main characteristics of Moslem medical institutions such as guilds, hospitals, and organizations of professional practice also are discussed. Although Arab medicine essentially constituted a transmission vehicle of master ideas of ancient medical thought, this medicine awoke the interest and initiative of the medieval physicians of western Europe, for example, those at the medical school of Salerno. PMID- 12096399 TI - [Superior neo-cava with autologous pericardium. Usefulness of radioisotope superior vena cavography]. PMID- 12096400 TI - [Opportunities for the pharmaceutical industry at the Mexican Genomic Medicine Institute]. PMID- 12096401 TI - [Laboratory animals and official Mexican norms (NOM-062-ZOO-1999)]. AB - This article concerns animal experimentation and official Mexican norm Nom 0062 Zoo-1999 entitled Technical specifications for the production, care and use of laboratory animals. The history of animal experimentation is briefly resumed. During the nineteenth century, doubts arose as to the right to expose animals to experimental procedures that frequently cause pain and suffering. The first law which protected animals against cruelty was passed in Great Britain in 1876; subsequently, other nations approved similar legislation. During the second part of the twentieth century, opposition to animal experimentation grew. Other groups, mainly scientists and pharmaceutical concerns, defended the right to use animals in research. New knowledge concerning the neurophysiology, cognitive capacity, and the animal faculty to experience pain is briefly mentioned. Guidelines on care and use of animals used in research published in several countries are listed. Finally, the recently published Mexican legislation (Norm) referring to production, care and use of laboratory animals is discussed and its benefits are stressed. PMID- 12096402 TI - [Academy and Medicine 2002]. PMID- 12096403 TI - Hearing controversy. GPOs testify to Senate subcommittee. PMID- 12096404 TI - Bundles of joy. PMID- 12096405 TI - Order in numbers. PMID- 12096406 TI - The business of compliance. PMID- 12096407 TI - Stats. Impacting equipment pricing. PMID- 12096408 TI - House GOP mulls major Medicare reform. PMID- 12096409 TI - CMS calls for 2.75 percent FY'03 inpatient PPS increase. PMID- 12096410 TI - Reps. Moran, Turner & McInnis introduce rural packages. PMID- 12096411 TI - The heart and soul of America's hospitals. PMID- 12096413 TI - Rebels in the pews. PMID- 12096412 TI - Modernizing Medicare: there is a better way. PMID- 12096414 TI - Reviving truth and trust. PMID- 12096415 TI - Who says the church can't change? PMID- 12096416 TI - A kiss before sneezing. PMID- 12096417 TI - Take me to your leader. Headhunters find few management mentors. PMID- 12096418 TI - It's easier to tell the truth. Legislation would allow protected error-reporting. PMID- 12096419 TI - VA announces capital program. PMID- 12096420 TI - Seeking consensus. Dissent leads AMA to scrap membership changes. PMID- 12096421 TI - Looking into a new method. AHA wants Joint Commission to try out volunteer teams from hospitals. PMID- 12096422 TI - When good PR goes bad. North Shore-Long Island Jewish gets a crash course in media management. PMID- 12096423 TI - Serving the mentally ill. Providers offer specialty ERs for psych care. PMID- 12096424 TI - Maine racks up another win. High court advised not to hear case against law. PMID- 12096425 TI - Parity has legs. With Bush on board (sort of), mental health act looks like a winner. PMID- 12096426 TI - Liable to be at risk. Absent tort reform, doctors, patients face fallout from malpractice verdicts. PMID- 12096427 TI - Cashing in on a crisis. The malpractice liability insurance crunch has proved to be a boon for temporary physician-staffing firms. But even those doctors aren't immune from the pain of rising premiums. PMID- 12096428 TI - Paying for the privilege. Some healthcare service awards are raising eyebrows among competitors, who question validity and application fees. AB - Hospitals hunting for a new marketing edge have found it--awards. The latest is the Choice Award, sponsored by an alliance formed by Ric Parr (left). This new focus on highlighting performance and service through laurels reflects a widening trend in consumer-driven healthcare. PMID- 12096429 TI - GPOs' draft conduct code faces little guys, lawmakers. Working document stays mostly silent on issues of product bundling and sole-source contracting. PMID- 12096430 TI - [Immunologic characteristics of the thymus of aged CBA mice after implantation of human fetal tissue]. AB - The effect of human fetal tissues (hypothalamic, liver, spleen, adrenal, ovarian, testicle, uterine, prostatic, gut, placental cells) collected from the fetuses at the 16-22 weeks of prenatal development, on the cytological structure of the thymus of 2-years old CBA mice in 7, 14, 42 and 64 days after start of treatment has been studied using morphological and immunological methods. The data obtained suggest that the implantation of human fetal tissues does not stop totally, but postponed age involution of thymus. PMID- 12096431 TI - [The effect of vilon on the thymus and spleen in a radiation model of premature aging]. AB - Low dose ionizing radiation induced accelerated aging of the thymus and the spleen in rat. Treatment with a synthetic dipeptide vilon (Lys-Glu) inhibited partly this process. The data suggest that vilon is a candidate for geriatric research and practice. PMID- 12096432 TI - [Functional morphology of organotypic culture of spleens from rats of various ages exposed to vilon]. AB - A synthetic dipeptide vilon (Lys-Glu, 20 ng/ml) was added to a cultural medium of explants taken from rats of 3 days, 3 weeks and 2 years of age. The peptide induced morphologic stability of the tissue and activated the regeneration and functional activity of cells. The stronger effect on the explants taken from the old rats suggests that vilon is a candidate for geriatric research and practice. PMID- 12096433 TI - [Diffusion ability of lungs at advanced age]. AB - Changes in lungs ventilation is mainly studied in the research on the features of respiratory system in ageing process. The condition of alveolar--capillary oxygen diffusion expressing the presence of central acinar lung emphysema is much less investigated. The purpose of the research was to study particularities of lung diffusion ability, which is considered to be one of the important mechanisms determining blood oxygenation in elderly. Results of the research and discussion. The dynamic of body surface changes is much less variable with age, than the dynamic of the lung diffusion ability. This is considered to be the reason of preservation of age differences of lung diffusion ability referred to 1 M2 of a body surface. There are two differences in dynamic of DLCO (diffusion lungs capacity) referred to total lungs capacity and referred to 1 M2 of a body surface. The indicator of double correlation of vital capacity of lungs (VCL) of elderly patients and Dlco makes +0.764; between intrathoracic gas volume and DLCO +0.654; between the residual volume of lungs (RV) and DLCO +0.729; between the Tiffno index and DlCO +0.654. The same parameters of young patients with COPD were 0.112; -0.245; 0.105; 0.312 correspondingly. Age related reduction of ratio of lungs diffusion ability to total lungs capacity is less prominent comparing to the ratio of DlCO body surface. The sex dependant differences of DlCO/TC smooth out in with each decade in contrast to DlCO/m2. A close correlation was revealed between DLCO and vital lungs capacity, the residual volume of lungs, intrathoracic gas volume and Tiffno indicator in elderly patients with COPD. PMID- 12096434 TI - [Contribution of I. V. Davydovskiy in the development of gerontology]. AB - This article represents a historical review concerning a period of work of prominent Russian pathologist Prof. I. Davydovski on a field of gerontology. At the end of his professional activity I. Davydovski has organized a gerontological laboratory. His research works have been summarized in the popular monograph "Gerontology", which appeared in 1966. This book served as a guidance in general and systemic gerontology for many years. PMID- 12096435 TI - [Social theory of aging in the USA: development and current status]. AB - Social theories of aging provide the basis of all relevant practice interventions, policies, programs and research. They evolve and change over time, based upon new knowledge gleaned from additional experience and research, as well as a more enlightened understanding of populations-at-risk and the psychological and social conditions that affect the well-being of such older adults. It is important to assess theories critically in order to use them judiciously. This abstract develops social theories that have evolved in the United States since the early 20th century to the present. They are presented in three main categories: Social Theories, Psychological Theories, and a combination of the two that recognizes the interaction between people and their social environments. Attention is also given to Spirituality and Aging, and to Biological considerations, particularly in regard to Sexuality and Aging. The authors contends that most theories are worthy of consideration, and none are necessarily applicable to everyone. The authors thank Prof. of social work Adelphi University Janice Wood Wetzel for the materials and help in writing this abstract. PMID- 12096436 TI - [Health status, functional prospects and use of medical and social services by the elderly living in St. Petersburg]. AB - The sharp increase of absolute number and share of the aged population in St. Petersburg requires changes in the medico-social service provision for the elderly. IPSE survey study was realized in 2000 in St. Petersburg under support of the Tacis Bistro Facility of the European Commission (The findings, conclusions and interpretations expressed in the article are those of authors and should not be taken to reflect the policies or opinions of the Europian Commission). The aim of the survey was to analyse health status, functional ability and use of medical and social services by the 60-89 aged elderly of the city and to identify the groups of elderly population in the most need of help. According to the results of the IPSE survey study, there is a significant number of elderly people who are not able to live independently at home without other people's help (approximately 97,000 elderly people in St. Petersburg who need assistance in daily living). The group of elderly people with considerably declined functional abilities should be a priority in planning the elder care services. For this group intervention programmes need to be developed. Providing the elderly with sufficient home help services fosters their independent living and allows the elderly to live longer at own home. To live at home is often the best choice for both the elderly and also the service system due to lower costs. PMID- 12096437 TI - [Way for improving the quality of life of elderly oncology patients]. AB - The study was concerned with the possibility to increase QL of cancer patients at older age. At our work we used lazeroauriculo-, acupuncture therapy in combination with psychotherapy. Results of tis treatment--74% good effect, 15%- statisfectory effect, 11%--without effect. Patients with advanced stage of cancer had a satisfactory effect only in 40% of cases. PMID- 12096438 TI - Thermodynamics of biological evolution and aging. Supramolecular thermodynamics is a key to understanding phenomena of life. What is life from a physical chemist's viewpoint. AB - The law of temporal hierarchies of the biological world allows us to pick out of the biomass quasi-closed thermodynamic systems with a given hierarchy. It has been established, that the use of this law of Nature as applied to supramolecular structures of organisms allows us the opportunity of using the methods of equilibrium supramolecular thermodynamics in the examination of open living systems. It has been shown that supramolecular thermodynamics is one of the "keys", which allows us to explain the origin of life and evolution of living beings. The second law of thermodynamics in its classic formulation (R. Clausius, J.W. Gibbs) is easy to apply in order to make calculations, carried out through methods of chemical, supramolecular and overall hierarchical thermodynamics. PMID- 12096439 TI - [Bio-physico-chemical aspects of aging and longevity]. AB - Currently two types of theories of aging are generally recognized: the genetic and the free radical theories. Each of them allow to explain plasibly some particular features of the aging process and pathologies associated with it. However, there are some phenomena related to aging and longevity that are not easily interpreted in frames of both theories; in particular, the increase of the life span of animals following caloric restriction, or beneficial effects of reactive oxygen species on multiple living functions, etc. On the other hand, the mostly unknown principles of theoretical biology suggested by E.S. Bauer in 1930s allow to explain from the monistic basis the essence of those and many other phenomena related to development, aging and longevity. The major postulates of the Bauer's theory and the corollaries of them are reviewed with the emphasis on the specifically biological phenomenon discovered by Bauer, the so called "Basic Process". The latter allows to find the means for the significant extension of the active life span. Taking Bauer's principles into consideration we contemplate the modern data on the peculiarities of free radical processes in living systems giving rise to electron excited species generation as the energy-informational foundation for the build up of an organism's resources for the counteraction of most pathological factors. The needfulness to employ these considerations for the resolution of the problems of gerontology is substantiated. PMID- 12096440 TI - [Aging of the pineal gland]. AB - The age-related changes in the pineal gland are functional rather than organic, which makes their correction or prevention more tenable. The amelioration or inhibition of some age-related impairments of the pineal gland were observed with dietary restriction and the use of S-adenosylmethionine or MAO-A inhibitors. A threefold increase in nocturnal melatonin peaks occurs in old rhesus monkeys treated with a synthetic peptide Ala-Glu-Asp-Gly (Epithalon) designed basing on the amino acid content of a pineal peptide extract Epithalamin. Other effects of Epithalon markedly overlap with melatonin effects. Besides life extension in mice and fruit flies, Epithalon effects include the postponing vision loss in Campbell rats with hereditary pigmental dystrophy. A uniting aspect of such a range of activities might be the participation of transcription factors, since they are often highly conservative in evolution and, on the other hand, may be strictly tissue-specific. The targets of Epithalon may include transfactors that in mammals are specific for the pineal gland and retina and exhibit impaired functions in the aged pineal gland. PMID- 12096441 TI - [Basic directions and prospects for international collaboration in gerontology. Role of the International Association of Gerontology]. AB - The overview presents data on the international collaboration in gerontology in the following spheres: 1) organizational basis of collaboration; 2) coordination of research; 3) coordination in the training of specialists; 4) collaboration in the organization of geriatric services; 5) collaboration in social care for the elderly. Role of the International Association of Gerontology and other international organizations in the promotion of gerontology and geriatrics in each of these spheres is shown by the example of Russia. PMID- 12096442 TI - [Significance of changes in individual indicators on intravascular homeostasis in development of circulatory hypoxia in aging]. AB - The study comprised 45 healthy subjects aged 60-74. It was designed to examine the state of skin and bulbar microcirculation using the method of television capillaroscopy. The functional state of endothelial microvessels was studied with the help of laser doppler flowmeter by means of functional tests with postocclusion hyperaemia and intradermal injection of epinephrine solution in the dose of 0.1 ml (1.10-6 mol/l). The platelets aggregation activity was studied using the method of turbidimetry. The blood viscosity and functional state of erythrocytes were determined with the help of rotary viscosimeter at the shift velocity of 10 to 200s-1 with estimating the erythrocytes deformability and aggregation indexes. Oxygen saturation of the subcutaneous fat fibers was studied by polarography. It has been found that the increase of adrenalin-induction aggregation of platelets, the decrease of erythrocyte deformation and the endothelium synthesis disturbance caused relaxing factors play role in the circulatory hypoxia development. PMID- 12096443 TI - [The retina and aging]. AB - The survey offers an analysis on the modern concepts of involutionary changes in the retina and the causes of age-related pathology occurrence in the eye. PMID- 12096444 TI - [Diabetes mellitus in the elderly: succinic acid compounds in treating diabetic neuropathies]. AB - To investigate the influence of the succinic acid treatment on geriatric patients with type 2 diabetes. Succinic Acid has some positive biological properties. One of its is a neglecting of an aerobic glycolysis. In this study we evaluated the efficacy of the combination of the succinic acid ("MITOMIN") on treating of diabetic neuropathy of geriatric patients with type 2 diabetes. The analysis was carried out using 26 patients (aged 60-76 years). The duration of diabetes was 9.15 +/- 1.43 years. Biomedical parameters were measured by standard methods; microalbuminuria was measured by "Micral-Test". Quality of life (psychosocial disorders) was estimated with the help of "SANDOZ"-scale for geriatric assessment. The therapy was assigned 1.5 g of mitomin per day during a month. All patients were examined on having late diabetic complications: 7.69%--had diabetic retinopathy; 11.54%--diabetic nephropathy; 73.08%--diabetic neuropathy; 46.15%- chronic failure of brain vessels; 11.5%--macroangiopathy of lower extremities and 100%--had ischeamic heart disease of different levels. Mitomin therapy improved basal and postprandial glycemic control (NS), variance of pallesthesia (p < 0.001), parameters of quality of life, i.e. depression (p < 0.001), anxiety (p < 0.01), short memory (p < 0.05) and emotionality (p < 0.001). Mitomin therapy plays a positive role in management of elderly patients with type 2 diabetes. It improves glycemic control, pallestesia and quality of life. Combination of succinic acid renders central and peripheral neuropathy protective efficacy. PMID- 12096445 TI - [Chemotherapeutic aspects of treating tumors in elderly patients]. AB - The importance of the problems of the choice for the treatment of elderly patients with solid tumors is demonstrated in the article. It is clear, that the age must not be reason for refusal in conducting therapy with cytostatics. The analysis of age changes in various organs and systems has been carried out as well as the influence of this changes on pharmacykinetics and pharmcodynamics of cytostatics. Practical recommendations have been given concerning chemotherapy in elderly patients. PMID- 12096446 TI - [Tissue-specific action of peptides in tissue culture of rats of various ages]. AB - There was studied the effect of polypeptide preparations isolated from the cerebral cortex (Cortexin), epiphysis (Epithalamin), liver (Hepalin), thymus (Thymalin) and small synthetic peptides--Cortagen, Epitalon, Livagen, Vilon on the development of explants from the cerebral cortex, brain subcortical structures, liver and thymus of rats of various age. With respect to the applied concentrations of the investigated peptides, they exerted tissue-specific effects in the organ-typical culture of animal tissues. PMID- 12096447 TI - Adolescent accounts of the UK National Lottery and scratchcards: an analysis using Q-sorts. AB - The study examined adolescents' accounts of the UK National Lottery and scratchcards. Q-sorts were used to examine the views of 62 participants aged between 11 and 15 years of age. Findings identified four distinct accounts in relation to the National Lottery (Moral Opposition, Luck Seeking, Rationalist, & Uncertainty), and four distinct accounts in relation to scratchcards (Scepticism, Thrill-Seeking, Rationalist, & Libertarian). Some of the accounts identified described the UK National Lottery and scratchcards as bona fide forms of gambling. Reports indicated that adolescents were pessimistic about the chances of winning large sums of money, while other accounts demonstrated gambling misperceptions particularly in relation to their belief in luck and the laws of probability. It is argued that to fully understand why adolescents take part in these activities it is important to consider the diverse ways that adolescents represent these activities. These differing representations will have consequences for measures aimed at reducing, preventing, or treating adolescent problem gambling. The utility of Q-sorts as a technique for examining the views of problem and non-problem gamblers is also discussed. PMID- 12096448 TI - A profile of gaming machine players in clubs in Sydney, Australia. AB - New South Wales, Australia, is one of the major markets in the world for machine gaming, with its 1,441 registered clubs holding the dominant share of the state's machines. This study examined machine gambling behavior amongst a random sample of 3,000 club members from six of the largest clubs in Sydney, the capital city of New South Wales. The findings identify their machine gambling preferences and participation levels, isolate significant sociodemographic and other differences between gaming machine and non-gaming machine players, measure the prevalence of probable pathological gambling associated with machine gambling, and identify certain characteristics which differ significantly between probable pathological and non-pathological machine gamblers amongst the respondents. PMID- 12096449 TI - The effect of compliance-improving interventions on the cognitive-behavioural treatment of pathological gambling. AB - This exploratory study investigated the effect of interventions designed to improve compliance and reduce dropout rates during the outpatient treatment of pathological gambling at a University-based gambling treatment clinic. Forty subjects (29 males, 11 females, mean age = 37.6) meeting DSM-IV criteria (APA, 1994) for pathological gambling were randomly assigned to either a cognitive behavioural treatment or a cognitive-behavioural treatment combined with interventions designed to improve treatment compliance. Compliance was indicated by the completion of all treatment sessions. Outcome measures were DSM-IV criteria assessed by structured clinical interview, South Oaks Gambling Screen scores, and percentage of income gambled. Logistic regression analyses identified pretreatment characteristics predicting compliance and outcome. Compliance improving interventions significantly reduced dropout rates, resulting in superior outcomes at posttreatment compared to the cognitive behavioural treatment alone. At 9-month follow-up, there was no difference in outcome between treatments, although both produced clinically significant change. Comorbid problem drinking, drug use, and problem gambling duration predicted poor compliance. Poor outcome was predicted by comorbid problem drinking. The clinical implications of these results are discussed in light of the exploratory nature of the study and the need for future research to address compliance, outcome, and comorbidity issues. PMID- 12096451 TI - [Antibiotics in abdominal surgical infections]. PMID- 12096452 TI - [Antibacterial agents: current status and perspectives]. PMID- 12096453 TI - [Formulary system in antibiotic therapy]. PMID- 12096454 TI - [Innovative approaches to the development of novel antibacterial agents]. PMID- 12096450 TI - The prevention of gambling problems in youth: a conceptual framework. AB - Despite increased awareness of the need to begin educating young children about the potential dangers of gambling, empirical knowledge of the prevention of adolescent problem gambling and its translation into science-based prevention initiatives is scarce. This paper poses the question of whether or not the common elements of tobacco, alcohol, and illicit drug abuse prevention programs can be applied to gambling prevention. Common risk and protective factors across addictions, including gambling, appear to point to the need to develop a general model of primary, secondary, and tertiary prevention. The authors present the need for science-based prevention initiatives and describe a general adolescent risk-taking model as a basis for science-based prevention of adolescent problem gambling and other risk behaviors. PMID- 12096455 TI - ["ABOLmed"--a Russian cephalosporin-manufacturing company]. PMID- 12096456 TI - [Infectious endocarditis in injection drug users]. PMID- 12096457 TI - [Ecology of drug resistance]. PMID- 12096458 TI - Facelift in a bottle. PMID- 12096459 TI - Is this artery about to blow? PMID- 12096460 TI - [Hematogenous (suspected-)septic inflammation of the synovial fossa in the adult horse: rare but deceptive (3 case reports)]. AB - Septic inflammation of the synovial fossa in the horse is a severe disorder that can lead to permanent invalidity. The synovitis can occur as a result of a perforating trauma, or via the blood (haematogenous). The latter route is rare in the adult horse. Three cases of suspected haematogenous septic synovitis are described in adult horses (older than 1 year). In two horses, timely and adequate treatment resulted in recovery, but this was followed by fatal recurrence. In one of these horses a persistent primary infection was found as cause; the other horse was not available for autopsy. The third horse was euthanized without treatment because the primary process was difficult to treat. It can be concluded that, in horses with suspected haematogenous septic inflammation of the synovial fossa, attention should be paid to the primary process--it is not enough to treat the synovitis alone. PMID- 12096461 TI - [Objectivity in the research of Gleptosil-Dextran]. PMID- 12096462 TI - [Anesthetic guns]. PMID- 12096463 TI - [Hysterectohysteria]. PMID- 12096464 TI - [Bladder infection with Capillaria plica in a male dog]. AB - In a male dog with symptoms of haematuria an infection with C. plica was established. During the first urine examination large numbers of erythrocytes and leucocytes were seen. The dog was treated with antibiotics. After 10 days a second urine examination revealed several eggs of C. plica, accompanied by leucocytes and erythrocytes. The infection was eliminated with fenbendazole treatment (50 mg/kg/day, for 10 days). PMID- 12096465 TI - [Scientific symposium at the occasion of the farewell of Dr. J.H.M. Nieuwenhuijs]. PMID- 12096466 TI - [Streptococcal infection is no zoonosis!]. PMID- 12096467 TI - [Clinical judgement for exhibitions (breeding, etc.) of cows, sheep and goats]. PMID- 12096469 TI - [European congress about aging of the brain in dogs]. PMID- 12096468 TI - [Waltham Award for evaluation of laparoscopic ovariectomy in the bitch]. PMID- 12096470 TI - [How do you make a diagnosis and what is the treatment of deep eye infections?]. PMID- 12096471 TI - Registered nurses key to good patient outcomes, study finds. But national nursing groups disagree over ratio laws and how best to recruit and retain quality nurses. PMID- 12096472 TI - Shedding few tiers. Controversial plans are here to stay, insurers say. PMID- 12096473 TI - A successful operation. Modern Healthcare's annual Hospital Systems Survey finds organizations are healthier operationally, but at not-for-profits, the gains don't always show up on the bottom line. PMID- 12096474 TI - Davidson and O'Leary talk to each other--and Modern Healthcare. Interview by John Morrissey. AB - The heads of the American Hospital Association and the Joint Commission on Accreditation of Healthcare Organizations waged a war of words this spring. At issue was the JCAHO's plan to seek a contract that the AHA contended would push the commission into the role of a regulator. Modern Healthcare broke the story about the disagreement. Now, Richard Davidson and Dennis O'Leary tell Modern Healthcare in an exclusive interview that they have patched up their differences. PMID- 12096476 TI - [Application of new FISH techniques for cytogenetic analysis in hematological malignancies]. PMID- 12096477 TI - [Gene expression analysis of AML cells using cDNA microarray]. PMID- 12096475 TI - [Cytocidal effects of gemtuzumab ozogamicin (CMA-676) on CD33-expressing acute myeloid leukemia cells, analysis P-glycoprotein and CD34 expression]. PMID- 12096478 TI - [Chromosomal translocations revealed by long-distance polymerase chain reaction targeting genomic DNA]. PMID- 12096480 TI - [Stratification of myeloma patients according to prognostic factors]. PMID- 12096479 TI - [Establishment of a quantitative method for detecting mixed chimerism using fluorescence-based PCR amplification of short tandem repeat markers after allogeneic hematopoietic stem cell transplantation in a Japanese population]. PMID- 12096481 TI - [Effect of autologous peripheral blood stem cell transplantation for multiple myeloma: a multi-institutional survey]. PMID- 12096482 TI - [Allogeneic stem cell transplantation with non-myeloablative or reduced-intensity conditioning for multiple myeloma]. PMID- 12096484 TI - [Effect of bisphosphonate on myeloma bone disease]. PMID- 12096483 TI - [Thalidomide for the treatment of refractory multiple myeloma]. PMID- 12096485 TI - [Monoclonal antibody therapies for acute leukemia]. PMID- 12096486 TI - [Mini-transplant as an immunotherapy]. PMID- 12096489 TI - [Antigen-presenting cells in cellular immunotherapy]. PMID- 12096488 TI - [Peptide-induced cytotoxic T lymphocytes]. PMID- 12096487 TI - [Cellular immunotherapy of hematologic malignancies attempted using minor histocompatibility antigens as targets]. PMID- 12096490 TI - [CTL and helper epitopes in DNA vaccines]. PMID- 12096491 TI - [Granulocytic sarcoma developing in lymph nodes]. AB - A 77-year-old man was admitted to a hospital because of a left cervical tumor. He was initially diagnosed as having non-Hodgkin lymphoma, diffuse large cell type, Ann Arbor stage IV, and transferred to our hospital for chemotherapy. Flow cytometric analysis of the left axillary lymph node cells derived from a biopsy specimen showed that in addition to lymphoid surface markers (CD5, 7, 21), myeloid surface markers (CD11b, 33, 34) were also positive. The diagnosis of malignant lymphoma was therefore confirmed. The patient, was treated with THP-COP therapy, which proved very effective. Thereafter, a biopsy specimen was found to be positive for MT1 (CD43) staining but negative for myeloperoxidase and chloroacetate esterase staining on immunohistochemistry. Furthermore, no rearrangement of the IgH JH, TCR C beta 1 or TCR J gamma gene was detected by Southern blot analysis. On basis of these findings and the previous results of flow cytometry, we changed the diagnosis from malignant lymphoma to granulocytic sarcoma. THP-COP therapy was continued, and complete remission was achieved. Two months later, however, the patient developed acute myelocytic leukemia (AML M1) and received DCP therapy, but he died of pneumonia. PMID- 12096492 TI - [Primary subcutaneous mantle cell lymphoma treated successfully with THP-COP therapy]. AB - A 73-year-old man noticed a subcutaneous tumor on the left upper palpebra from April 1998, but did not seek therapy for it. Facial subcutaneous tumors appeared from November 1999, and multiple tumors appeared on the skin of the chest and both upper arms from January 2000. Tumor biopsy revealed that these tumors were non-Hodgkin lymphoma showing CD19 (+), CD20 (+), CD5 (+), CD10 (-), smIgM (+), sm lambda (+) and cyclin D1 (+). The karyotype was t(11;14) (q13;q32), but bcl-1 gene rearrangement was not detected. On the basis of these data, primary mantle cell lymphoma (MCL) of the subcutis was diagnosed. The patient underwent eight courses of THP-COP therapy, and complete remission was achieved. Primary subcutaneous B-cell lymphoma, especially MCL, is rare. MCL is aggressive and difficult to cure; the median survival of patients is 3 to 5 years, and the 5 year survival is 30%. However, the present patient showed a good response to chemotherapy, and complete remission has continued for 17 months since the MCL was first diagnosed. PMID- 12096493 TI - [Acute myelogenous leukemia complicated by compartment syndrome]. AB - A 37-year-old woman was diagnosed as having acute myelogenous leukemia. After obtaining complete remission by induction therapy, consolidation therapy was performed. She collapsed during an episode of chemotherapy-induced pancytopenia and developed compartment syndrome of the lower leg a week later. Fasciotomy was performed successfully, and she maintained her complete remission. Early diagnosis and fasciotomy are important in patients with leukemia complicated by compartment syndrome. PMID- 12096494 TI - [Effects of pulmonary rehabilitation in patients with pulmonary tuberculosis sequelae]. AB - Pulmonary rehabilitation was evaluated for a mean period of 3.9 weeks in 37 inpatients with pulmonary tuberculosis sequelae. The rehabilitation program consisted of relaxation, breathing retraining, exercise training, respiratory muscle training and instruction. Significant improvement was shown in VC (n = 37) on average from 1.48 l to 1.59 l, in FEV1.0 (n = 37) from 0.93 l to 1.02 l, in PaO2 (n = 35) from 67.1 Torr to 72.4 Torr, in 6-minute walking distance (n = 29) from 303 m to 339 m, in Pimax (n = 17) from 38.5 cmH2O to 47.5 cmH2O, in activity (n = 23) from 19.6 points to 22.5 points, in dyspnea (n = 22) from 18.4 points to 22.5 points and in QOL (n = 25) from 39.0 points to 44.2 points. The effects of pulmonary rehabilitation did not depend on past thoracic surgery for tuberculosis, pattern of ventilatory impairment, findings of chest radiography, or degree of insufficiency. These data suggest that pulmonary rehabilitation is of benefit for improving pulmonary function, exercise tolerance, symptoms and QOL in patients with pulmonary tuberculosis sequelae. PMID- 12096495 TI - [Clinical characteristics in 8 sporadic cases of community-acquired Legionella pneumonia]. AB - To study the clinical characteristics of community-acquired Legionella pneumonia, we carried out a study of 8 patients (7 men and a woman; mean age, 68.4 years) with Legionella pneumonia at Yaizu Municipal General Hospital between 1996 and 1999. We surmised that the sources of infection were travel or a hot spring, or both, in 4 cases, occupation (plumbing and fish market work) in 2 cases and gardening in one. All patients had fever above 38 degrees C, hypoxemia and a high inflammation reaction of. The initial findings of chest radiography were air space consolidation in all cases, lobar pneumonia in 7, and unilateral shadows in 6, similar to those seen in acute Streptococcus pneumoniae pneumonia. Diagnostic methods were urinary antigen in 4 of 7 cases, bacterial culture in 4 of 6 cases (L. pneumophila, 3 cases; L. longbeachae, 1 case), polymerase chain reaction on the serum in 2 of 5 cases, and serum antibody in 1 of 7 cases. Urinary antigen was most useful for early diagnosis. The clinical presentation and the initial chest radiography findings were non-specific, despite the high fever, severe hypoxemia, and radical progression within a few days. PMID- 12096496 TI - [Evaluation of cardiopulmonary function in Japanese Himalayan climbers to the Himalayas underin a hypobaric hypoxic chamber]. AB - The Tokai University and Tibet University Scientific Friendship Expedition 2001 succeeded in reaching the summit of Kula Kangri (7,538 m) in the Himalrayas without supplementary O2 in May, 2001. Before this expedition, the cardiopulmonary function of all members was evaluated using a hypobaric hypoxic chamber in which the barometric pressure was gradually lowered to a simulated altitude of 6000 m. Whenever a member's PaO2 was lower than 40 torr during the experiment, supplementary O2 was given. The results were divided into two groups, Group A and Group B, according to whether supplementary O2 was used. Group A subjects did not need O2 even at an altitude of 6,000 m, whereas Group B subjects did when they reached an altitude of 5,000 m. Seven the eight Group A members had climbed mountains over 4,000 m several times, whereas twelve out of the thirteen members of Group B had no mountaineering experience. The mean PaO2 and PaCO2 of Group A at 6000 m were 41.9 torr and 31.9 torr, respectively, while those of Group B at 5000 m were 35.8 torr and 35.9 torr. There was a significant difference in the degree of hyperventilation between Group A and Group B. No difference was observed between the two groups in systolic and diastolic arterial blood pressure, but the heart rate in Group B increased significantly under hypoxia. These results suggest that the experience of climbing mountains over 4000 m may lead to significant differences in cardiopulmonary function under hypoxia. PMID- 12096497 TI - [A case of lymphomatoid granulomatosis with multiple thin-walled cavities]. AB - We report a case of rapidly progressive lymphomatoid granulomatosis. A 48-year old man was admitted because of hemoptysis and high fever. A chest radiograph and chest computed tomograms revealed multiple nodules, thin-walled cavities in the lungs, some containing ball-like masses. After admission, clinical studies, including percutaneous and transbronchial biopsies failed to provide sufficient evidence for proper diagnosis. Fluoro-deoxyglucose positron emission tomography (FDG-PET) showed strong accumulations of abnormal shadows in the lesions. A VATS biopsy was performed to make possible an exact diagnosis. The histological findings showed angiocentric lesions with infiltration of polymorphous cells, and were compatible with lymphomatoid granulomatosis (LYG). The respiratory failure progressed as the reticular shadows in the chest radiograph increased, so treatment with methylprednisolone pulse therapy (1,000 mg/day for 3 days) was started, and followed with prednisolone therapy (60 mg/day for 3 weeks). Initially, the symptoms improved, but gradually deteriorated, and the pulmonary nodules became enlarged. The patient finally died of progressive respiratory failure in addition to uncontrollable empyema with an MRSA infection. We attributed the cavity formation and ball-like masses in the cavities to the occlusion of small pulmonary arteries and tissue necrosis along the drainage bronchus. PMID- 12096498 TI - [A case of fatal MALT lymphoma of pulmonary origin with systemic metastasis and manifesting macroglobulinemia]. AB - The patient was a 61-year-old man whose chest radiography showed infiltration in both lung fields. MALT lymphoma was diagnosed from transbronchial lung biopsy (TBLB) specimens. The serum monoclonal IgM level then increased, and plasma cells and lymphoplasmacytoid cells were found in a bone marrow examination, indicating macroglobulinemia as a complication. Despite chemotherapy, systemic metastasis led to the patient's death. Our report is accompanied with a review of literature, because no report of MALT lymphoma of pulmonary origin complicated with macroglobulinemia exists, and it is considered that the macroglobulinemia in this case was related to the MALT lymphoma. PMID- 12096499 TI - [A case of miliary tuberculosis complicated by acute respiratory failure after bronchofiberscopy]. AB - A 79-year-old man presented with persistent nonproductive cough and high fever. The chest radiograph showed bilateral miliary shadows. We performed bronchoalveolar lavage and transbronchial lung biopsy to assist diagnosis. Severe dyspnea developed after the bronchofiberscopy, when the chest radiograph revealed bilateral ground-glass shadows and the oxygen saturation in the room air fell to 60%. It was suspected that the patient had acute respiratory distress syndrome, so that methylprednisolone was given intravenously at a dose of 250 mg daily for 3 days, which resulted in a reduction in ground glass shadows and an improvement in oxygen saturation. We diagnosed miliary tuberculosis because the transbronchial lung biopsy specimen showed caseous granuloma and the PCR test for Mycobacterium tuberculosis in the bronchoalveolar lavage fluid was positive. The patient was cured with antituberculosis chemotherapy. PMID- 12096500 TI - [Occluding spiral embolus for treatment of refractory pulmonary fistula]. AB - A 57-year-old man was admitted for pulmonary emphysema. Volume-reduction-surgery (VRS) was performed. Three days later, after he coughed hard, the right lung collapsed. Although surgery was repeated, the air leakage continued. Air leakage was immediately reduced by means of bronchial embolization with a spiral embolus. We suggest that bronchial embolization with a spiral embolus causes little harm and is an effective treatment for refractory pulmonary fistula in poor-risk patients. PMID- 12096502 TI - [Pulmonary eosinophilic granuloma in a woman of 72]. AB - We report a case of pulmonary eosinophilic granuloma with the latest onset ever reported in Japan. A 72-year-old woman, a current smoker, developed dyspnea and dry cough in late January 2001. Chest radiography and CT examinations revealed numerous small nodular lesions and multiple cysts in both upper lung fields. A thoracoscopic lung biopsy revealed a granulomatous lesion composed of eosinophils and large histiocytic cells with pale eosinophilic cytoplasms and large cleaved nuclei. These cells gave a positive reaction for S-100 protein. These findings led to a diagnosis of pulmonary eosinophilic granuloma. This disease is generally reported in much younger subjects, and few cases are known in those over 60-years old. No Japanese report of histologically confirmed pulmonary eosinophilic granuloma in an older patient has previously appeared, but it may now be prudent to take this disease into consideration when diagnosing pulmonary disease in older subjects. PMID- 12096501 TI - [Pulmonary sarcoidosis in a case of dermatomyositis under long-term steroid therapy]. AB - Pulmonary sarcoidosis in association with dermatomyositis has rarely been reported. A 52-year-old woman was admitted for evaluation of a previously unrecognized bilateral hilar lymphadenopathy on a chest radiogram in August 1995. She had been receiving oral corticosteroids for dermatomyositis, which was confirmed by the typical skin rash and the myogenic changes on an electromyogram in 1977. She had no respiratory symptoms at the time of admission. Chest computed tomography showed hilar and mediastinal lymphadenopathy with reticular shadows in the peripheral lung parenchyma. A 67gallium-citrate scintigram apparently revealed accumulation in bilateral hilums. The tuberculin skin reaction was negative. Flowcytometry of bronchoalveolar lavage fluid indicated a CD4/CD8 ratio of 5.37. Thoracoscopic biopsy specimens sampled from the mediastinal lymph nodes and the lung demonstrated non-caseating granulomas. A definitive diagnosis of pulmonary sarcoidosis was made from these findings. The chest radiographic findings were spontaneously resolved after three years with no increase in the dosage of oral corticosteroids. It is well known that an abnormal immune response playa an important role in development of dermatomyositis and sarcoidosis. This case suggested close associations in both diseases. PMID- 12096503 TI - [A case of pulmonary infiltration with eosinophilia (PIE) syndrome induced by bucillamine treatment of rheumatoid arthritis]. AB - Bucillamine is used mainly in treating rheumatoid arthritis (RA). We report a case of bucillamine-induced pulmonary infiltration with eosinophilia (PIE) syndrome in a 51-year-old woman. When RA was diagnosed, she was treated with bucillamine from December 2000. In April 2001, she was admitted to our hospital because of fever and skin eruptions. Chest radiography and CT revealed both diffuse ground-glass opacity and fine nodular shadows. Laboratory data showed a normal white cell count with eosinophilia. Bronchoalveolar lavage (BAL) studies showed that total cell counts and the proportion of eosinophils were increased, and that the CD4/CD8 ratio of the T-cell subsets was decreased to 0.93. The patch test to bucillamine was positive. After bucillamine was withdrawn, the fever and the abnormal chest shadows improved. We concluded from the patient's clinical course, laboratory data and BAL findings that this was a case of bucillamine induced PIE syndrome. Since most cases of bucillamine-induced interstitial pneumonitis are lymphocytic alveolitis, we consider that PIE syndrome in such a case is a very rare condition. We concluded that bucillamine should be added to the list of drugs capable of producing PIE syndrome. PMID- 12096504 TI - [Improvement of extrapulmonary lesions of eosinophilic granuloma after cessation of smoking]. AB - We encountered a case of pulmonary eosinophilic granuloma complicated with pneumothorax. A 24-year-old man was admitted to our department because of respiratory difficulty. Chest radiography on admission showed a right pneumothorax. A thoracic catheter was therefore inserted, and the pneumothorax was improved. A chest radiograph obtained after treatment showed diffuse linear reticular shadows; and a chest CT scan showed starlike nodular lesions and multiple cysts in the lungs, and a large nodule in the left rib. Because an ulcer had been diagnosed in the oral cavity 1 year previously, a mandible biopsy was performed, and a granulomatous lesion consisting of eosinophils was recognized. Taken together with the pulmonary findings, an eosinophilic granuloma was diagnosed. The patient was instructed not to smoke and the clinical course was observed. The pulmonary, mandibular, and costal lesions improved. PMID- 12096505 TI - [A long-term surviving patient with invasive thymoma who underwent radiotherapy and/or resection for chest wall, intrathoracic and intrapelvic recurrent tumors]. AB - A 49-year-old woman with myasthenia gravis who underwent left panpleuropneumonectomy for an invasive thymoma that disseminated through the left thoracic cavity. After six year, radiotherapy was conducted on the recurrent tumor in the left anterior chest wall. Two years later, the recurrent tumors in the intrapelvic and intrathoracic cavities were resected. It was thought that long-term survival was obtained by combining radiotherapy and surgical treatment in view of the patient's general condition, and of the recurrent invasive thymoma present in this case. PMID- 12096506 TI - [A case of desmoplastic malignant mesothelioma with elevated serum CYFRA 21-1]. AB - A 70-year-old woman was admitted to our hospital for medical evaluation of a right side pleural effusion, which was pointed out at another hospital. Chest CT revealed a right pleural effusion with diffuse and irregular pleural thickening. Percutaneous pleural biopsy showed hypocellular collagenous tissue without malignant cells. Though she received antituberculosis therapy, the pleural thickening progressed and the serum CYFRA 21-1 level was elevated. Chest pain and dyspnea appeared, and she was readmitted. However, pneumonia was present as a complication, and she died. At autopsy, the right pleura was thickened and invasion of the lung and the chest wall was observed. Microscopic findings showed increased amounts of hyalinized collagen fibers forming a storiform pattern. At the tumor foci, atypical cells with distinct nucleoli were observed. Desmoplastic malignant mesothelioma, which is rarely reported in Japan, was diagnosed. PMID- 12096507 TI - Battle of the faithful. Catholics are voicing hurt and anger over the church's sexual-abuse crisis. Is the hierarchy listening? PMID- 12096508 TI - So berry good for you. PMID- 12096510 TI - Not-for-profits in credit crunch? Think tank's warning comes as capital needs rise. PMID- 12096509 TI - Data show operating margins take step up. PMID- 12096511 TI - Small group takes on Tenet. Latino group's suit alleges patients were overcharged. PMID- 12096512 TI - Preserving quality of education. Accreditation Council limits residents' work week. PMID- 12096513 TI - Study: Docs contribute to nurse shortage. PMID- 12096514 TI - Too much money? Executive compensation issue in CareFirst conversion. PMID- 12096515 TI - Making a case for innovation. Boston system's new center to spread word on IT. PMID- 12096516 TI - An attractive model. Hospital bets on unique financing plan to save itself. PMID- 12096517 TI - That board seems awfully small. St. Francis blames change on inadequate coverage. PMID- 12096518 TI - Rocky D.C. campaign. Push for hospital a monumental effort. PMID- 12096519 TI - Breaking the bank. Suit says doc owner drove hospital into bankruptcy. PMID- 12096520 TI - CFOs to the rescue. After Enron and AHERF, financial oversight is playing a far more pivotal role. PMID- 12096521 TI - Web survey. May results: 'boutique medicine'. PMID- 12096522 TI - So much, yet so little. When it comes to financial data on not-for-profit hospitals, there are plenty of snapshots, but it's still hard to see the big picture. PMID- 12096523 TI - Wait and see. Lengthy delay in Medicare cost reports puts industry analysis on hold. PMID- 12096524 TI - Joint ventures win the draw. Judge's ruling to revive tax exemption for Texas system spells good news for investor-owned systems, not-for-profits seeking deals. AB - A ruling by a federal judge reinstating the tax exemption of St. David's Health Care System in Austin, Texas, has taught the Internal Revenue Service a hard lesson about the toughness of Texas. No one was happier with the judge's decision than St. David's President and Chief Executive Officer. PMID- 12096525 TI - [Tactics of treating children with Coat's disease]. AB - Fifty children with Coats disease, aged 2 months to 12 years (mean age 8.25 +/- 2.72 years), were observed. Four stages are distinguished in the disease course: initial, moderate, advanced, and terminal. Treatment including extensive retinal argon laser coagulation, cryotherapy, scleral bucking, and subretinal liquid draining was carried out in 32 children (33 eyes). Stable anatomic results were attained in 97% eyes. Visual acuity of at least 0.02 was retained in 87.9% children. PMID- 12096526 TI - [Treatment of lacrimal duct diseases using low energy helium-neon laser]. AB - Treatment of diseases of the lacrimal duct remains a pressing problem in ophthalmology. These diseases are responsible for 71-85.4% diseases in capable people. Sixty patients with chronic dacryocystitis with partially retained patency of the lacrimal duct were treated by He-Ne low-frequency laser ULF-01 (output power 6.10 mcWt, laser beam wave length 0.632 nm). The patients received 3-5 min sessions twice a week, 5-8 sessions per course. Positive effect was attained in 56 patients: complete cessation of excessive lacrimal discharge in 38 patients and subjective improvement in 18. He-Ne laser exposure brings about a good antiinflammatory effect; in combination with antibiotic therapy it promotes rapid sanitization of the lacrimal duct, removes edema, and rapidly normalizes lacrimal discharge. PMID- 12096527 TI - [Actipol in treating stromal herpetic keratitis]. AB - Actipol (0.007% paraaminobenzoic acid--PABA) is a new interferon (IFN) inductor. It was recently introduced into ophthalmological practice. Its efficiency in surface herpetic keratitis is proven. We studied the therapeutic efficiency of actipol in the treatment of stromal herpetic keratitis and compared the results with combined therapy with acyclovir (ACV) and leukocytic IFN. The main group (141 patients) were treated with actipol and the reference group (40 patients) with ACV ointment and leukocytic IFN. Local injections of actipol in combination with its instillations into the conjunctival sac led to cure of 67.3% patients with stromal herpetic keratitis; this treatment was more effective than combined local ACV + leukocytic IFN therapy (clinical cure in 45% cases). Epithelialization in the actipol group was observed 2 days sooner, infiltration resorption and clinical cure 4 days sooner than in the reference group. Relatively high visual acuity in the actipol group was presumably due to the reparogenic effect on the corneal stroma and antithrombotic, fibrinolytic, and antioxidant activity of PABA. Hence, actipol is an effective drug for the treatment of stromal herpetic keratitis, exerting virtually no side effects. PMID- 12096528 TI - [Pathological basis for clinical stages of the proliferative process in diabetic retinopathy]. AB - In order to elucidate the relationships between posterior ocular structures, 48 samples of epiretinal membranes were examined, which were obtained during transcilliary vitrectomy for proliferative diabetic retinopathy of different stages (29 patients with type I diabetes mellitus and 19 with type 2). The findings of morphological analysis and preoperative ophthalmoscopy (color photographs) demonstrated clinical and morphological correlations of stages of proliferative diabetic retinopathy. Impossibility of differential diagnosis between posterohyaloid membrane and newly formed tissue by clinical methods and impossibility of their mechanical intraoperative separation allow us to regard the stages of proliferative process as stages in changes of the posterohyaloid membrane proper. The posterohyaloid membrane is detected clinically and morphologically only in the presence of pathological changes, including diabetic retinopathy. We detected the following clinical histological correlations of proliferative process stages in diabetic retinopathy: 1) glial; 2) glio-vascular; 3) glio-vascular-fibrous; 4) fibrovascular; 5) fibrous (cicatricial). The histology of stages of proliferative diabetic retinopathy and correlation of histological findings with the clinical (ophthalmoscopic) picture of the process are described in detail. Recommendations on the treatment of each of these stages are offered. PMID- 12096529 TI - [Features of the clinical course and treatment of some types of complications in the early postoperative period after intrastromal laser keratectomy]. AB - Some complications observed during the early postoperative period after intrastromal laser keratectomy by LASIK and REIK are described: diffuse interface infiltration, flap "ripples", central dystrophy of the flap, infective keratitis. Differential diagnostic signs of diffuse infiltration in the interface, central dystrophy of the flap, and infective keratitis are described and data on the clinical physiological status of the cornea (fluorescein penetrability, lacrimal production tests) in these conditions are presented. The efficiency of drug therapy is evaluated and protocols for the treatment of these complications are presented. PMID- 12096530 TI - [Comparative results of sub-flap intrastromal photokeratectomy in correcting astigmatism by bitoric and monotoric keratoablation]. AB - The paper discusses the results of 28 laser operations carried out by bitoric nomograms and 18 operations carried out by monotoric nomograms for correction of high astigmatism (3 and more diopters) by laser in situ keratomileusis (LASIK). The patients' ages varied from 21 to 38 years. Eximer photokeratectomy was carried on a EC 5000 Nidek laser, lamellar section of the cornea was made with a Hansatome microkeratome. Bitoric keratoablation was carried out by an original method. Clinical refraction, visual acuity, and incidence of reoparations were evaluated in both groups 6 months postoperation. Differences in refraction and visual acuity were negligible. The incidence of reoparations in bitoric keratoablation was 25%, in monotic one 44%. Visual acuity with correction was higher after bitoric keratoablation of astigmatism. Hence, bitoric keratoablation is preferable for correction of high astigmatism by LASIK. PMID- 12096531 TI - [Diagnosis of post-traumatic uveitis using ultrasonic dopplerography]. AB - Changes in the brain biopotentials during early and remote periods after perforating wounds of the eyes were for the first time investigated in 55 patients with posttraumatic uveitis (PTU). Increased alpha-rhythmic activity of the brain was detected, its location and intensity depending on the severity of injury and PTU and course of PTU. The possibility of predicting the outcome of uveitis was demonstrated and diagnostic value of the method in evaluation of its severity was proven. In favorable outcomes of PTU, alpha-rhythm intensified in the anterior compartments of the brain. In severe uveitis and risk of sympathetic inflammation, brain activity increased in the posterior compartments presenting as sharp waves, being pronounced in both hemispheres or on the intact eye side. PMID- 12096532 TI - [Role of antiphospholipid antibodies in occlusion of retinal vessels in various vascular eye diseases]. AB - A strong association between the presence of antiphospholipid antibodies (aPLab) and arterial and venous thrombosis was observed during the past decade. aPLab represent a heterogeneous group of immunoglobulins that include the lupus anticoagulant (LA), anticardiolipin antibodies (aCL), and antibodies responsible for false-positive test for syphilis. aPLab react with negatively charged, rarely with neutral phospholipids and/or phospholipid-binding proteins. aPLab are associated with vascular occlusion in autoimmune diseases, e.g. systemic lupus erythematosus (SLE). The role of aCL in ocular vascular occlusive disease involving retinal vessels is unclear. The aim of this study was to determine the relationship between aPLab and occlusive retinal disease. A relationship between aCL (IgG), LA, and thrombotic retinal events has been detected in SLE patients, but no association between aPLab and other occlusive retinal diseases (central retinal vein thrombosis, isolated retinal vasculitis, optic nerve vasculitis). In our study the incidence of aCL IgG was higher in patients with Behcet disease with non-occlusive thrombosis than in patients with retinal occlusive events or in patients without occlusions or thrombosis. It seems that different mechanisms are responsible for these conditions. PMID- 12096533 TI - [Causes, prognosis and prevention of cicatrical corneal astigmatism]. AB - Penetrating wounds of the eyeball in 57-66% cases are associated with corneal involvement leading to the development of cicatricial astigmatism and decrease of the functional results. We investigated the causes of posttraumatic cicatricial corneal astigmatism, possibilities of its prediction, prevention, and correction. The results of treatment of 256 patients with corneal wounds and cicatrices are analyzed. Our analysis confirmed that the severity of astigmatism in corneal wounds directly depends on the wound size, edge status, direction, location, and structure of the forming cicatrix. Adequate and timely primary debridement of the corneal wound with intraoperative keratometric control plays the key role in minimization of cicatricial astigmatism. Early therapy with drugs stimulating the regeneration processes promotes more complete recovery of corneal tissue structure and decrease of astigmatism severity. Timely dosed slackening or removal of sutures makes possible modulation of astigmatism development. A formula is proposed, using which makes it possible, on the basis of initial data on the corneal wound, to estimate the possibility of corneal astigmatism, i.e. predict the outcome of the injury. PMID- 12096534 TI - [Status of the cardiovascular system in patients with normal pressure glaucoma]. AB - Twenty patients with normal pressure glaucoma (NPD) were examined. Concomitant cardiovascular diseases were diagnosed in 19 (95%) of them. Echocardioscopy with color mapping showed various diseases with left-ventricular dysfunction in 8 examinees. Dopplerography of orbital vessels showed a significant decrease of bloodflow velocity in the orbital artery and increased peripheral resistance indexes in the orbital and central retinal arteries in 20 NPD patients in comparison with healthy controls (p < 0.05, p < 0.01). Atherosclerotic involvement of the internal carotid artery was detected in 15 of 20 patients, in 4 of these with hemodynamically significant sclerosis of more than 50%. The contribution of cardiovascular disease to decrease of orbital bloodflow is discussed. PMID- 12096535 TI - [Conservative therapy in rehabilitating patients with eye injuries]. AB - The authors offer recommendations on the choice and use of optimal clinically tried drugs in ophthalmic traumatology with consideration for posttraumatic changes, doses, and mechanism of action. The recommendations are based on the clinical observations of 16,000 patients treated at department of ocular injuries, reconstructive surgery, and ocular prosthetics of Helmholtz Institute of Ocular Diseases in Moscow. PMID- 12096536 TI - [Neuroophthalmic symptoms of multiple sclerosis]. AB - The significance of neuroophthalmological symptoms in the diagnosis of disseminated sclerosis are analyzed. Fifty-two patients presented with the optic form of the disease of different severity. Subclinical stage of contralateral involvement was detected in patients with unilateral retrobulbar neuritis, which indicated dissemination of the process. Diseases of optic and oculomotor nerves of unknown etiology in young subjects can be an early symptom of disseminated sclerosis. The frequency of ocular and oculomotor disorders in disseminated sclerosis dictates regular neuroophthalmological examinations of patients with suspected disseminated sclerosis for early detection of the disease, which is particularly important for prediction of its course and outcome. PMID- 12096537 TI - [Features of cataract surgery in children and adolescents]. AB - The postoperative period following extracapsular cataract extraction in children and young people is as a rule characterized by opacities of the posterior capsule of the lens and fibrosis of anterior borderline vitreous membrane, requiring reoperations. The key factor in prevention of reoperations in surgery for congenital cataracts and cataracts of different etiology in children and youths is primary posterior capsulorrhexis with anterior vitrectomy. Posterior capsulorrhexis without anterior vitrectomy is recommended for patients aged over 14 years, in whom vitreous fibrosis is less probable. PMID- 12096538 TI - [Increase in visual acuity in poorly-sighted patients by using spectral filters]. AB - Tinted eyeglasses were selected for 768 poor-sighted patients aged 5-86 years with various ocular diseases. The frequency of tinted glasses utilization by poor sighted patients was as follows: filter O2 was the optimal for 38% patients, filters G2 and K4 for 16-18%, and G1 for 11% patients. Filter G1 improved visual acuity by 19%, filters G2 and K4-K5 by 23%; the highest increment of visual acuity was observed with O2 filter: by 27%. Filters of different spectral characteristics and density may be useful in the same underlying ophthalmic disease, which is explained by the underlying disease and the presence, combinations, and severity of concomitant ocular diseases. PMID- 12096539 TI - [Uveitis, caused by a wasp bite]. PMID- 12096540 TI - [The problem of decompression the optic nerve in traumatic neuropathies]. PMID- 12096541 TI - [Neuro-reflexive apparatus of the cornea as a cause and effect factor in developing retinal pathology and complications after photorefractive surgery]. PMID- 12096542 TI - [Idiopathic macular rupture: pathogenesis, diagnosis, and treatment]. PMID- 12096543 TI - [An express-method for predicting fibrinous iridocyclitis in the early postoperative period in artiphakia]. AB - The authors investigated the possibility of using the level of preoperative blood monocytosis as an express method for predicting fibrinous iridocyclitis in planned artiphakia. Extracapsular cataract extraction with intraocular correction (B. N. Alexeev's design) was carried out in 372 patients, 287 of these without non-insulin-dependent diabetes mellitus (NIDDM) (group 1) and 85 with NIDDM (group 2). The patients were divided into 3 subgroups depending on the absolute blood monocyte counts before operation: 1) up to 200 monocytes, 2) 200-400, and 3) more than 400. The incidence of fibrinous iridocyclitis was significantly higher (p < 0.01, 17-18-fold on average) in both subgroups 3 (more than 400 monocytes) in comparison with subgroups 1 (less than 200 monocytes), the treatment strategy being the same in all patients. In NIDDM group the total number of fibrinous iridocyclitis cases was 2 times higher in all 3 subgroups than in group 1. Hence, the level of preoperative blood monocytosis can be used as an express method for predicting fibrinous iridocyclitis in planned artiphakia choosing the treatment strategy during the immediate postoperative period, including that for patients with NIDDM. PMID- 12096544 TI - [Dependence of mobility of an eye prosthesis on the method of rectus muscle fixation]. AB - The relationship between the mobility of eye prosthesis and method of musculus rectus fixation was investigated. The results indicate that fixation of the muscles to subconjunctival tissues allows the best mobility of the eye prosthesis. This method of musculus rectus fixation is most similar to anatomical fixation. In addition, fixation of musculus rectus is essential for configuration and mobility of the conjunctival cavity. Hence, the optimal method for fixation of musculus rectus in order to attain the best mobility of the eye prosthesis and therefore the best cosmetic result can be chosen during enucleation. PMID- 12096546 TI - Getting patients the support they need. PMID- 12096545 TI - Quality of life: healing the brain and the body. PMID- 12096547 TI - Necrotizing fasciitis: a diagnostic and management challenge. PMID- 12096548 TI - The patient with skin disease: an approach for nondermatologists. AB - Dermatologists and other healthcare professionals frequently encounter patients with skin disease in their daily practice. Developing an approach to these patients that will lead to accurate and timely diagnosis is important for all healthcare providers. Increased awareness of the potential benefits and pitfalls of a dermatologic history, knowledge of the fundamentals of a dermatologic examination, the ability to provide a potential differential diagnosis, and improved communication with dermatologists will improve care of patients with skin disease. PMID- 12096549 TI - Sacral skin blood perfusion: a factor in pressure ulcers? AB - Pressure ulcers occur over the sacrum but are rare over the gluteus maximus. This predilection is partly explained by pressure concentration effects, but other factors may be involved. The authors hypothesized that if resting sacral skin blood flow was greater than in surrounding tissues, a decrease or stoppage of blood flow during loading might represent increased risk because relative tissue deficits would be greater. Because information on sacral skin blood flow is scarce yet important to the plausibility of this hypothesis, the objective was to determine if actual sacral skin resting blood flow is, in fact, different than flow in other nearby posterior areas. Thus, skin blood flow was measured with laser Doppler imaging in 15-cm2 areas overlying the sacrum in 30 subjects (15 male) and compared to skin blood flow in other posterior sites (gluteus maximus and lower back) and to remote sites (hand and fingers). Results showed that average sacral skin blood flow (59.1 +/- 1.4 arbitrary profusion units) was significantly (P < 0.001) greater than other posterior sites (48.7 +/- 2.5 a.u.) and was significantly (P < 0.01) greater in females (63.0 +/- 1.6 vs. 55.2 +/- 1.8). These findings provide the first systematic characterization of resting sacral skin blood flow. The data are consistent with the tentative hypothesis, but more direct evidence of a linkage is clearly needed. Large spatial variability in skin blood flow (40%) suggests that comparisons of skin blood flow among sites are best done with laser Doppler imaging in contrast to standard laser Doppler monitoring. PMID- 12096551 TI - Experts. PMID- 12096550 TI - Clinical and cost effectiveness of a cleanser protectant lotion for treatment of perineal skin breakdown in low-risk patients with incontinence. AB - Perineal dermatitis due to urinary and/or fecal incontinence is a common problem. A multicenter, open label, phase II product evaluation was conducted to determine the effectiveness of a new cleanser protectant lotion in reducing perineal erythema and pain in patients at low-risk for perineal dermatitis and to compare the cost of this product to standard protocols of care. Nineteen elderly patients (14 male, 5 female, mean age 73.1 years) participated in the study. Average baseline scores for erythema and pain were 2.3 (+/- 0.5) and 1.5 (+/- 1.0), respectively (scale 0 to 4). After 7 days, both scores were significantly lower (mean scores 0.6 +/- 0.8 and 0.3 +/- 0.8, respectively; P < 0.01). Based on an average of 2.6 perineal episodes per day, the one-step product evaluated would cost $136 per patient/year less than standard protocols of care while reducing caregiver time (average 23 seconds per episode of care). Optimal perineal care may reduce the incidence of complications; studies to ascertain the safety and effectiveness of commonly used products and procedures are needed. PMID- 12096552 TI - Reimbursement updates. PMID- 12096553 TI - Assessing youth/adult partnerships: the Seven Circles (AK) experience. AB - Seven Circles is a substance abuse prevention coalition in Southeast Alaska with a two-tiered structure consisting of local partnerships joined together in a regional coalition. Seven Circles incorporates a youth/adult partnership approach to accomplish its activities. This article describes the results of four annual administrations of a partnership member survey designed to assess the development and implementation of youth/adult partnerships in the local projects, assess the value of the partnerships to the participants, and gather feedback about the functioning of the Seven Circles Coalition. The results support the youth/adult partnership model and are also discussed in the context of youth empowerment. PMID- 12096554 TI - Initial findings from Parent Party Patrol: an intervention to reduce adolescent substance use through reduced involvement in unchaperoned parties. AB - This article presents preliminary evaluation findings from Parent Party Patrol (PPP). PPP is a two-hour single-session intervention that provides information to parents about ways to increase monitoring and improve communication and family guidelines regarding substance use, with the goal of reducing adolescent involvement in unsupervised activities and associated substance use. Seventy three attendees from 15 PPP sessions completed baseline and three- to six-month follow-up information. At follow-up, participants showed significantly increased awareness of adolescent substance use and unchaperoned activities, improved communication and use of family guidelines regarding substance use, and increased monitoring of their children's activities. A significant reduction in adolescent attendance at unchaperoned parties was also reported. Replication of the study with a larger, more representative sample and controlled design is suggested. As adolescent substance use is a multidetermined problem, PPP may be best viewed as one part of a broader comprehensive substance abuse prevention program. PMID- 12096555 TI - Measurement of risk and protective factors for drug use and anti-social behavior among high school students in South Africa. AB - The study sought to examine, for South African adolescents: 1) the reliability of sub-scales of the Communities that Care Youth Survey (CTC Youth Survey) of risk and protective factors for drug use and anti-social behavior; and 2) the extent to which tobacco, alcohol and marijuana use can be predicted from community, family, school, and peer-individual factors based on sub-scales of the CTC Youth Survey. On two occasions, 92 male and 31 female, Grade 8 and 11 students completed measures concerning: 1) their past month tobacco, alcohol and marijuana use; and 2) various community, family, school, and peer-individual factors. Cronbach alpha coefficients of sub-scales of the questionnaire ranged between .60 and .94. Kappa values were at least moderate (above .40) on 19 sub-scales, and on the remaining sub-scales observed agreement levels ranged between .49 and .94. Each domain predicted tobacco, alcohol, and marijuana use. Multiple logistic regression analyses revealed that alcohol use was most strongly accounted for by the peer domain, tobacco use by the school domain, and marijuana use by the peer and community domains. The findings support use of the CTC Youth Survey, with slight revisions, among South African high school students. PMID- 12096556 TI - Alcohol misuse among college athletes: self-medication for psychiatric symptoms? AB - A collegiate athlete population was surveyed for alcohol abuse as well as self reported depression, anxiety, and other psychiatric symptoms. This study revealed that in a group of 262 athletes there were 21 percent who reported high alcohol use and problems associated with its use. Significant correlations were found between reported alcohol abuse and self-reported symptoms of depression and general psychiatric symptoms. Subjects with positive depression and psychiatric symptom ratings in the "severe" range had a significantly higher rate of alcohol abuse than subjects who had low depression and low or mild symptom ratings. Conversely, subjects reporting higher rates of alcohol misuse had more psychiatric symptoms. These findings suggest a possible causal link between psychopathology and serious alcohol abuse among college athletes. They also point to the need for routine depression and anxiety screening in college students who are typically beginning a significant exposure to alcohol. PMID- 12096557 TI - An assessment of drug testing within the construction industry. AB - Drug testing in the workplace has gone from virtual nonexistence to widespread employer acceptance during the past two decades. This growth is particularly significant for the construction industry. High rates of alcohol and other drug use, coupled with the high-risk, safety-sensitive nature of the industry, have prompted the development of a variety of drug surveillance and prevention strategies. Despite this growing vigilance, no scholarly works have examined the impact of drug-related policies in the construction industry. To address this limitation, we investigate the efficacy of workplace drug-testing programs in reducing injury incident rates and workers' compensation experience-rating modification factors (MODs) within the construction industry. Analyses indicate that companies with drug-testing programs experienced a 51 percent reduction in incident rates within two years of implementation. Moreover, companies that drug test their employees experienced a significant reduction in their MODs. Policy implications are discussed in light of the current findings. PMID- 12096558 TI - Structural equation modeling of drinking motives in male DUI offenders. AB - Measures completed by 250 men convicted of driving under the influence of alcohol (DUI) were analyzed to examine the direct effects of parental problem drinking and the mediational influence of drinking motives on alcohol use and drinking related problem outcomes. Structural equation modeling was used to evaluate competing models in the cross-sectional data set. Results indicated that: 1) reported parental problem drinking modestly predicted both alcohol use and problem outcomes when tested in the absence of influence from drinking motives; 2) confirmatory factor analysis yielded a 4-factor drinking motives solution consisting of social, enhancement, coping with negative emotions, and interpersonal facilitation; and 3) both enhancement and interpersonal facilitation drinking motives mediated parental problem drinking's direct effect on alcohol use and problem outcomes. Drinking to cope with negative emotions, while not mediational in nature, produced a direct effect pathway to problem outcomes. The findings extend predictive utility of specific drinking motive dimensions on alcohol use and problem outcomes from previous research in community and university samples to men convicted of driving while intoxicated. PMID- 12096559 TI - Age differences in alcohol and cocaine expectancies and attitudes. AB - Positive and negative expectancies regarding the behavioral effects of alcohol and cocaine were assessed and used to predict attitudes toward their use across four age groups (5-7, 8-10, 11-14, and 18-25, N = 121). Regardless of gender and minority status, children and early adolescents appeared to overgeneralize their beliefs about alcohol to a less familiar drug, cocaine, perceiving the effects of the two drugs similarly. Only college students differentiated between drugs, perceiving cocaine as less likely than alcohol to produce drunkenness and more likely to have stimulant and elation/empowerment effects. With age and other expectancies controlled, expectancy of drunkenness was the best predictor of disapproval of alcohol use; attitudes toward cocaine use were unrelated to expectancies but became more negative with age. Drug prevention programs should rest on data regarding children's preexisting beliefs about the consequences of drug use and should help them understand that different drugs (for example, stimulants and depressants) pose different dangers. PMID- 12096560 TI - Therapeutic work with children in pain. PMID- 12096561 TI - Enhancing seamless care: a review. PMID- 12096562 TI - Is a CCN degree the only way to develop children's nursing in the community? AB - Semi-structured interviews were carried out with convenience samples 10 CCN students completing a community children's nursing (CCN) degree programme and with five practice educators. In addition, focus group interviews were held with 28 child branch students who had undertaken community experience. The objective was to find out the perceived value of undertaking a community children's nursing degree. The overall aim of the study was to address practical questions of teachers who ran the CCN pathway within the community degree programme and inform programme development. The data confirmed ENB and UKCC views of the CCN role and supported the need for more flexible approaches to CCN education. Recommendations are made for changes, particularly in relation to the core community content of the programme which did not meet students' expectations for developing their child specialist role within a multi-agency team. PMID- 12096563 TI - Children's books: a resource for children's nursing care. PMID- 12096564 TI - Preventive mental health. PMID- 12096565 TI - Respite care for children who have complex healthcare needs. PMID- 12096566 TI - The child first and always: the registered children's nurse over 150 years. Part Two. PMID- 12096567 TI - Standards for perinatal education. Part 2. PMID- 12096568 TI - Breastfeeding practices of working women. AB - Breastfeeding is the fundamental aspect of child rearing that a woman copes with. It creates a wonderful bond between mothers and their babies. The importance of breast feeding to child health is highlighted by the yearly World Breastfeeding week (Forste, Weiss & Lippincott 2001:291). Breastfeeding promotions in the Northern province like anywhere else in the country appear effective and visible enough to encourage women at all levels to breastfeed. Ironically, many women give up breastfeeding when they return to work because the thoughts of expressing their milk at work and the fiddling around with pumps and storage at work seem overwhelming (McAlpine 1998:80). Based on the assumption that breastfeeding requires commitment and determination, the researcher thought it was necessary to enquire about the experiences of working women on breastfeeding. Focus group interviews were conducted using a semi-structured format. Twenty-six women participated in three focus group discussions. The study was conducted in the suburban areas of the Northern Province, Soutpansberg region. Data was transcribed from a taped session and analysed using an open code method. Four major categories were identified: Knowledge and attitudes towards breastfeeding, lack of support at work, lack of support at home and a need for facilities and resources that promote breastfeeding at the workplace. It was perceived that women are so stressed at the end of the day that they have little or no time to nurse the baby. They appear however knowledgeable about the benefits, advantages and disadvantages of breastfeeding their babies. A concern was also raised that the time that they therefore spend with their babies is limited and affect their relationship with their babies. Women felt that they need support from their employers as well as colleagues at work to be able to balance their responsibilities. PMID- 12096569 TI - Changing stress levels through gaining information on stress. AB - OBJECTIVE: The aim of this research was to find out the effect of the Information Phase of a Stress Management Program (SMP) on the perceptions of participants about their stress levels. METHOD: A total sample of 100 workers (nursing staff, private business men and women, laboratory assistants, the protective services [foreman and security staff], as well as people in human resources departments) took part in this study. All the participants were from the Northern and Gauteng Provinces in South Africa. The Combined Hassles and Uplifts Scale (Folkman & Lazarus, 1989) was used as an instrument to measure the perceived stress level of participants in a SMP. RESULT: A significant reduction in stress levels was achieved among those who received the Information Phase of the SMP only, as well as those who received the whole stress management techniques. There was no significant difference between the amount of reduction in perceived stress-levels achieved among those that received the Information Phase of the SMP only, compared to that of those who received the whole techniques. CONCLUSION: The authors conclude that where the resources are limited, only the information phase of a SMP may be given to desiring clients. That should help to save time and money spent on participating in SMPs. This should however not discourage the use of the whole SPM, where affordable. PMID- 12096570 TI - A framework for caring in the human resource management process of nurses. AB - The aim of this paper is to describe the provision of a human value system in nursing management that would lead to an environment conducive to quality of care to nurses. Caring is a complex phenomenon with caring concepts which emphasized the human element in human resource management in the health services. Watson (1985) focused on the philosophical and spiritual basis of caring. The ten carative factors as described by Watson (1985) were used as a framework for the application of caring to the human resource management process in nursing. This framework enables a basis for the development of a caring environment for the nurse manager, in which the nurse could experience caring. It served as a point of departure for a research study undertaken during 2000 on caring to the caregivers in health services. PMID- 12096571 TI - Standards for perinatal education. Part 1. AB - Quality control in perinatal education is imperative, with the purpose to deliver a high quality and appropriate healthcare service to the family in the perinatal period. The purpose of this study was to develop valid standards for perinatal education in South Africa. To reach this goal, the study proceeded as follows: Development phase concepts were developed for perinatal education, the concepts were logically systematized and formulated in concept standards, the standards were presented for peer group review with the purpose of refinement and further development. Quantification phase the statistical validity of the standards was determined/calculated. After completion of the study the researcher reached the conclusion that quality perinatal education must adhere to the following standards: A philosophy for perinatal education must exist. The perinatal facilitator must be a well trained professional person, and must possess certain personal traits. The perinatal education style/method must adapt according to the client's socio-economic status, age, marital status and needs. The perinatal facilitator must make use of acknowledged teaching methods and techniques in order to facilitate learning. The perinatal education must comply with certain minimum requirements in order to make it accessible and acceptable to the client. The perinatal facilitator must complete a preparatory phase before she is allowed to give education. A perinatal education programme must have a curriculum that can be adapted to the client's circumstances. Perinatal education must be presented purposefully. The end result of perinatal education must focus on a healthy baby and a healthy, empowered mother, family and community. PMID- 12096572 TI - An investigation of the roles and functions of nurse preceptors in the clinical areas. AB - This article explores the confusion that exists in the clinical areas concerning proper accompaniment of student nurses. There is a feeling that professional nurses in clinical areas are not always actively involved in clinical teaching. The clinical instructors and nurse educators are unable to accompany student nurses properly because of staff shortages. Some hospitals have identified one professional nurse per unit to work as a nurse preceptor for accompaniment of student nurses. This has resulted in the need to find out if the professional nurse preceptor is solving the problem of lack of student accompaniment in clinical areas. The aim of this study therefore is to investigate the roles and functions of nurse preceptors in improving student accompaniment as compared to other professional nurses in clinical areas namely, clinical instructors, nurse educators and professional nurses working in clinical areas as viewed by student nurses themselves. A comparative descriptive study was done in one of the hospitals in KwaZulu-Natal (Region D) on a purposely selected sample of (80) 4 years comprehensive diploma student nurses, (16) nurse preceptors and (40) randomly-selected professional nurses. Open and closed-ended questions were used for collection of data. The study revealed that the most student nurses 87.5% (70) identified nurse preceptors as playing an important role in their accompaniment as compared to other professional nurses. Student nurses identified the following roles and functions of nurse preceptors in student accompaniment: acting as role models, acting as resource people for them, providing clinical teaching, orientate them in clinical areas, allay fears and anxieties by providing guidance, support and encouragement, demonstrate procedures, help them in solving problems they experienced and do formative and summative evaluation of student nurses. These roles were in agreement with those identified by professional nurses and nurse preceptors themselves. It was found also that nurse preceptors were experiencing many problems which need to be solved in order for them to perform their roles and functions effectively. It is recommended that even those hospitals and educational institutions that are not utilizing nurse preceptors for student accompaniment should consider them because of the contributions they make. Nurse preceptors should be allocated specifically for student accompaniment with minimal patient care responsibilities and administration of Units. In conclusion proper utilization of nurse preceptors can solve the problem of a lack of student accompaniment in clinical areas. PMID- 12096574 TI - Attitudes towards and beliefs about schizophrenia in Xhosa families with affected probands. AB - OBJECTIVE: The development of effective psychoeducational programs for the management of schizophrenia requires an understanding of attitudes towards and beliefs about the disorder in families of affected probands. In order to establish the need for adaptation of Western psychoeducational programs, these variables were investigated in Xhosa speaking families in South Africa. DESIGN: Xhosa speaking family members of patients with DSM-IV schizophrenia were recruited on a voluntary basis, and interviewed with a structured belief and attitudes questionnaire adapted from previous studies in the West. SETTING: The study population was drawn from both urban and rural Xhosa communities in South Africa. SUBJECTS: 100 Xhosa speaking family members participated in the study. RESULTS: Family members most often recommended treatment with psychotropic medications (88%) and traditional healers (32%), and least often recommended psychotherapy (4%) and meditation (1%). Of the respondents who recommended traditional healing methods, 92% also recommended simultaneous use of allopathic treatment. CONCLUSION: Attitudes towards and beliefs about schizophrenia in family members of patients with schizophrenia may differ substantially from those described in previous work in the West. An understanding of local attitudes and beliefs is crucial for the successful development of local psychoeducational programs. PMID- 12096573 TI - A value clarification on quality within nursing colleges in Gauteng. AB - Quality assurance in higher education has been legislated. It is therefore necessary to develop a quality audit system for Nursing Colleges in Gauteng. The process of developing such a system is complex and needs to foster ownership by all the stakeholders. The first step in this process is to conduct a value clarification on quality. The purpose of this article is to explore and describe a value clarification on quality within the Nursing Colleges in Gauteng. An explanatory and descriptive qualitative research design was utilised. The results are grouped into structure, process and outcome values. Comparisons between the views of the different role-players are also presented. It is recommended that these results be utilised as the basis of a conceptual framework for the quality audit system for Nursing Colleges in Gauteng, as well as the development of quality indicators for Nursing Education Institutions in South Africa. PMID- 12096575 TI - A comparative analysis of pressure sore treatment modalities in community settings. PMID- 12096577 TI - Aeromonas hydrophila wound infection in elective surgery. AB - Patients with diabetes mellitus are subject to wound infection with unusual pathogens. A patient who had colonic surgery developed Aeromonas hydrophila wound infection, which mainly results from contamination by water or soil. PMID- 12096576 TI - Inflammation. 1: Its role in the healing of acute wounds. AB - The inflammatory response is an essential part of the wound-healing process. This article discusses inflammation in acute wounds and the part played by the migration of cells including neutrophils, macrophages and T-lymphocytes. PMID- 12096578 TI - Mouseyes revisited: upgrading a computer program that aids wound measurement. AB - A computer program (Mouseyes) for measuring wounds has been upgraded to incorporate the use of photographic images taken with a digital camera. This evaluation showed that the upgrade produced accurate and reproducible results. PMID- 12096579 TI - Sterile saline versus tap water: which is most effective? PMID- 12096580 TI - Cost-effectiveness of wound management in France: pressure ulcers and venous leg ulcers. AB - OBJECTIVES: This study set out to define realistic protocols of care for the treatment of chronic venous leg ulcers and pressure ulcers in France and, by developing cost-effectiveness models, to compare the different protocols of care for the two ulcer groups, enabling a calculation of direct medical costs per ulcer healed in a typical French health insurance plan. METHOD: Clinical outcomes and some treatment patterns were obtained from published literature. Validations of different treatment patterns were developed using an expert consensus panel similar to the Delphi approach. Costs were calculated based on national averages and estimates from the UK and Germany. The models were used to measure costs per healed ulcer over a 12-week period. RESULTS: For both the pressure ulcer and venous leg ulcer models, three protocols of care were identified. For pressure ulcers and venous leg ulcers, the hydrocolloid DuoDERM (ConvaTec, also known as Granuflex in the UK and Varihesive in Germany) was most cost-effective in France. CONCLUSION: The combination of published data and expert consensus opinion is a valid technique, and in this case suggests that treating pressure ulcers and venous leg ulcers with hydrocolloid dressings is more cost-effective than treating them with saline gauze, in spite of the lower unit cost of the latter. PMID- 12096581 TI - Assessing patients' nutritional needs in the wound-healing process. AB - Like all illnesses, successful wound management depends on appropriate nutritional support. This article outlines which nutrients are relevant to the various stages of wound healing and discusses the consequences of poor nutrition. PMID- 12096582 TI - Do continence aids help to maintain skin integrity? AB - The third of our back to basics series reviews the efficacy of continence aids such as pads and barrier creams, and their role in maintaining skin integrity. Many studies had flaws and new research is needed to avoid subjective choice of products. PMID- 12096583 TI - Urban infrastructure: the "out of sight, out of mind" mentality is an outmoded concept. PMID- 12096584 TI - A last chance. Can America's bishops end the sex-abuse crisis? PMID- 12096585 TI - Nursing wounds. When arrogant docs drive nurses away, patients suffer. PMID- 12096586 TI - How small a pox? PMID- 12096587 TI - Lightning lore to live by safely. PMID- 12096588 TI - Viruses from hell. PMID- 12096589 TI - When an HMO denial is practicing medicine. PMID- 12096590 TI - "We can take care of ourselves, dad". PMID- 12096591 TI - Charles G. Wagner, MD. Taking good works up, up, and away. PMID- 12096592 TI - Coping with your own child's illness. PMID- 12096593 TI - When a lawyer wants your records. PMID- 12096594 TI - Make sure you're paid for counseling. PMID- 12096595 TI - I'd rather give up ATMs. PMID- 12096596 TI - A $10 million allergy case. Could it happen to you? PMID- 12096597 TI - Details, details: what your colleagues think of drug reps. PMID- 12096598 TI - Why your patient won't let you touch her. PMID- 12096599 TI - From fingersticks to chopsticks. PMID- 12096601 TI - My patients are also my constituents. PMID- 12096600 TI - The importance of being curious. PMID- 12096602 TI - Running late. PMID- 12096603 TI - Confess and repent. PMID- 12096604 TI - Cracking the cancer code. PMID- 12096605 TI - Speak no evil. PMID- 12096606 TI - Sam's club. PMID- 12096608 TI - New hope for an ailing heart. PMID- 12096609 TI - The disappearing mind. PMID- 12096607 TI - The fatal promise of cloning. PMID- 12096610 TI - Was blind but now I see. PMID- 12096611 TI - How to recharge the second sense. PMID- 12096613 TI - Magnets that move moods. PMID- 12096612 TI - Healthy shocks to the head. PMID- 12096614 TI - Wiring the new docs. PMID- 12096615 TI - Your next.... PMID- 12096616 TI - The quest for artificial blood. PMID- 12096617 TI - Tiny weapons with giant potential. PMID- 12096618 TI - Unraveling the genome. PMID- 12096619 TI - A year later, the beat goes on. PMID- 12096620 TI - A day of atonement. PMID- 12096621 TI - [Studies of distribution of phosphorylated H3 in human MCF-7 cells by immunofluroescence labeling]. AB - One of the prominent cell cycle-related modifications of histone proteins whose function is correlated with chromosome condensation is the phosphorylation of histone H3. In this work we used immunofluorescence labeling on human MCF-7 cells with the antibody that was specific for phosphorylated histone H3 at Ser10 to examine the cellular distribution of this protein. The acid-soluble proteins from interphase and mitotic cells were separated by SDS-PAGE and the transferred proteins were probed with the antibody. A strong H3-specific band was only detected in the acid-soluble proteins from mitotic cells, demonstrating the correlation between H3 phosphorylation and mitosis. With confocal microscopy on whole cells, our results showed that mitotic phosphorylation of H3 initiated in discrete foci near the nuclear envelope in early prophase cells. Following initiation, H3 phosphorylation appeared to spread throughout the condensing chromatin and reached maximum in early metaphase cells. Dephosphorylation of H3 began in anaphase cells and was complete immediately prior to detectable chromosome decondensation in telophase cells. There was a precise spatial and temporal correlation between H3 phosphorylation and initial stages of chromatin condensation. The possible functions of the singular phosphorylation of the amino terminus of H3 were discussed. PMID- 12096623 TI - [The study of the uncoulping protein gene as the candidate gene for fatness traits in chicken]. AB - The UCPs are integral membrane proteins of the mitochondrial respiration from oxidative phosphorylation, diminishing the resulting production of ATP and instead yielding dissipative heat. The action of those proteins creates a futile cycle that decreases the metabolic efficiency of the organism. Thus UCPs provide new clue to obesity's causes. This study was designed to investigate the effect of UCP gene on chicken fatness traits. The fifth generation population of divergent selection broiler line, Hyline Brown layer and three native breeds (shiqiza, Beijing You, baier) were used in this research. Body weight and body composition traits were measured in broiler lines at 7 weeks of age. Primers for the 3'-untranslator region in UCP were designed from database of chicken genomic sequence. Polymorphisms were detected by PCR-SSCP and DNA sequencing. The results showed that there was significant difference (P < 0.01) in the frequency of genotype among breeds except broiler vs Beijing You and Baier vs Hyline Brown layer in mutation sites detected by the two pairs of primers. The distribution of genotype in Beijing You and broiler had no difference. It deduced that Beijing You belongs to the native breed that has dominant meat type traits and has the same genetic background with broiler. Baier and Hyline Brown Layer have no difference in the genotype, it can be viewed as they have same genetic background. A A/C mutation at base position 1197 was found among individuals in broiler line and the least square analysis showed that BB birds had significant lower (P < 0.01) abdominal fat weight and percentage of abdominal fat than AB or AA birds. From the results we can putatively draw the conclusion that UCP gene is the major gene to affect the fatness traits or it links with the major gene. PMID- 12096622 TI - [Mapping and expression analysis of a different expression cDNA fragment from lung adenocarcinoma cell line]. AB - Lung cancer is one of the most common malignant tumors in humans. Metastasis is the basic biological feature of malignant tumors, which is the main cause of death. Molecular mechanism of metastasis is still unclear, although lots of studies have been done in tumor metastasis. To study and explore the molecular basis of metastasis in lung cancer, and isolate tumor metastasis-related genes, two human lung adenocarcinoma cell lines AGZY 83-a and Anip 973 were chosen as research materials. The Anip973 was derived from AGZY83-a, but manifested much higher metastasis potential than the parent line. Using mRNA differential display technique, an unknown cDNA fragment, OPB7-1, which is over-expressive in Anip973 cell line, was obtained. It was used as a template to isolate its corresponding cDNA through dbEST searching and PCR. To search and clone lung adenocarcinoma metastasis-related candidate gene, and to explore the molecular basis of development of lung carcinoma, differential expression of OPB7-1 cDNA fragment among 9 human lung adenocarcinoma cell lines and 12 normal human tissues were detected using cell culture, cDNA clone, Northern blot analysis and bioinformation technology. Results showed that there were significant differences in OPB7-1 expression among 9 human lung adenocarcinoma cell lines. High expression tendency was observed in Anip973 cell line with high metastasis potential, TKB-18 cell line with high invasion potential and GLC-82 cell line with low differentiation potential. Besides, a bigger fragment can be found in Anip973 cell line on the Northern blot hybridization. The 3.0 kb transcriptions were found in various tissues. Over-expression in heart and skeletal muscle could be observed, whereas expression in spleen, liver, kidney, placental and lung could be found except colon, thyroid gland and small intestine. These manifests indicate that OPB7-1 gene has a wide-rage expression in human multiple tissues. A 1.0 kb cDNA fragment was acquired by linking up EST fragments homologous match 5' end and PCR. BLAST analysis revealed that OPB7-1 gene has extremely low sequence identity with any known genes from GenBank and any sequences from EST database. The chromosomal localization of it was determined by RH location method. The OPB7 1 fragment was localized to chromosome 1p31-34. That OPB7-1 gene has an extensive expression pattern, may be a novel tumor gene related to lung carcinoma. Further research needs to be done to obtain the full-length cDNA of OPB7-1 gene. It will be helpful to investigate the expression in lung cancer cases and other tumor tissues for further determining the function of OPB7-1 gene in development of tumor. PMID- 12096624 TI - [The selecting experiment of resistance to Marek's disease of chicken]. AB - The Marek's Disease is an acute infectious disease in poultry production. The selecting experiment through blood-group gene inspection of adult parents and attack-test by using of Marek's Disease virus (MDV) for offspring chicks was carried out. The results showed as follows: (1) The rates of positive reaction by No. 614 blood-group gene inspection of adult parents in four generations were increased due to generation increase, as F0-68.8%, F1-77.3%, F2-81.8% and F3 90.5%. (2) In the attack-test by using of MDV at 10 days of age of chicks the mortalities of the experimental group at 60 days of age of chicks whose parents have positive reaction by No. 614 blood-group reagent detect in F2-F4 generations were 25.8%, 69.2% and 29.4% respectively, while the mortalities of the control group at same age of chicks whose parents have negative reaction were 30.6%, 81.0% and 52.9% respectively. These differences of mortality were significant (P < 0.01 or P < 0.05). (3) The rates of pathological change of 9 viscera tumor of the experimental group chicks in F2-F4 generations were 87.6%, 83.3% and 58.8% respectively, while chicks in the control group were 88.1%, 84.5% and 70.6% respectively. Except individual generation and viscera, these differences between the experimental group and the control group were almost not significant (P > 0.05). The experimental results and genetic control of resistance to Marek's Disease are discussed in this paper. The new strain of chicken resistant to Marek's Disease will be bred. PMID- 12096625 TI - [The developmental patterns of GHr gene expression in gastric tissue of Erhualian and large white pigs]. AB - Pituitary growth hormone (GH) is essential for postnatal growth in animal, it regulates numerous cellular functions by direct effect on it's receptor (GHr) in many different tissues. And it is believed that the abundance of GHr in different tissue determines the tissue sensitivity to GH. However, the regulation of GH on porcine stomach development is still unknown. So, in present experiment single tube semi-quantitative RT-PCR was applied to investigate the developmental patterns of gastric GHr mRNA expression in Large White and Erhualian Boars, and the Classic 18S internal standards (Ambion Inc. USA) was used. Pigs were sampled at birth, and 3, 20, 30, 45, 90, 120, 180 days of age respectively, meanwhile the bodyweight and gastric weight were recorded. The results indicated that the expression of gastric GHr mRNA was high at birth, followed by a significantly decrease (P < 0.05) at 3 days of age (D3) in both breeds. In Erhualian boars, GHr mRNA expression reached a peak at D45, then remarkablly declined till D90 (P < 0.05), and kept constant thereafter. In Large White boars, however, the expression of GHr mRNA reached a peak at D90 and remained pleateau afterwards. From birth to D30, the GHr mRNA expression was higher in Erhualian than that in Large White, the significant strain difference were founded at D20 (P < 0.05). But from D30 to D180, the higher expression was found in Large white, the significant strain difference was especially at D90 (P < 0.05). Significant positive correlation was found between the expression of GHr mRNA and the ratio of gastric weight to bodyweight from birth to D180 (r = 0.54, P < 0.05). The results suggest that, (1) GHr mRNA in porcine stomach is expressed according a strain specific development pattern; (2) GH directly acts at the gastric tissue and regulates it's growth. The relationship between GH and the development of gastric function is to be elucidated. PMID- 12096626 TI - [Molecular screening of MC4R gene and association with fat traits in pig resource family]. AB - Melanocortin-4 Receptor (MC4R) plays an important role in the regulation of human obesity. It can cooperate with leptin, neuropeptide Y(NPY) and melanocyte stimulating hormone (MSH) to regulate body weight and feeding. Inactivation of this receptor by gene targeting in mice results in a maturity onset obesity syndrome associated with hyperphagic, hyperinsulinemia, hyperglycemia, as well as decreased linear growth and adult obesity. Multiple alignments of the sequences from individuals of several pig lines identified a single nucleotide substitution(G-->A) at position 298 of the seventh transmembrane domain. In present study, polymorphism distribution of MC4R gene fragment in resource population was studied using PCR-RFLP method based on the enzyme Taq I. The genotype was analyzed with the phenotype of the slaughtered individuals. The results showed that the frequencies of MC4R genotype varied in different breeds. The correlation analysis demonstrated the genotype of MC4R was in significant relation with back-fat thickness on thorax-waist, buttock and the average back fat thickness, as well as with the width and area of longissmus dorsi (LD), and the percentage of skin. MC4R gene plays a role mainly in the pattern of dominant effect, and all the additive effects were not significant. PMID- 12096627 TI - [Genetic polymorphisms of five microsatellite loci in Small Tail Han sheep]. AB - Small Tail Han sheep is an excellent local sheep breed in China. Small Tail Han sheep had significant characteristics of high prolificacy. The lambing percentage averaged 260 percent in Small Tail Han sheep. The polymorphisms of 5 microsatellite loci OarAE101, BM1329, BMS2508, TGLA54 and TGLA68 which were closely linked to the fecundity genes FecB and FecXI in sheep were detected in 244 ewes of Small Tail Han sheep. The PCR amplified products of microsatellites were detected by non-denatured (natural) polyacry lamide gel electrophoresis. Allele frequency, polymorphism information content, gene homogeneity and heterozygosity for 5 microsatellite loci were calculated. The number of alleles for BM1329, OarAE101, TGLA54, TGLA68 and BMS2508 were 6, 9, 5, 2 and 6 in Small Tail Han sheep, respectively. The range of allele sizes for BM1329, OarAE101, TGLA54, TGLA68 and BMS2508 were 160 bp to 180 bp, 97 bp to 135 bp, 116 bp to 136 bp, 98 bp to 100 bp, and 93 bp to 115 bp, respectively. The alleles of the greatest frequency for BM1329, OarAE101, TGLA54 and BMS2508 were 164 bp, 97 bp, 134 bp and 99 bp, respectively. Polymorphism information content/gene homogeneity/heterozygosity for BM1329, OarAE101, TGLA54, TGLA68 and BMS2508 were 0.4481/0.4840/0.5160, 0.3516/0.6375/0.3625, 0.2528/0.7326/0.2674, 0.3733/0.5034/0.4966 and 0.5809/0.3581/0.6419 in Small Tail Han sheep, respectively. The results revealed the greatest genetic variation in BMS2508 and the lowest in TGLA54. These results could provide, basic molecular data for the research on the germplasm characteristics of Small Tail Han sheep. PMID- 12096628 TI - [Genetic relationships among Oxya agavisa and other relative species revealed by Cyt b sequences]. AB - The mtDNA Cyt b genes (432 bp) were sequenced for 16 individuals of Oxya agavisa from 7 Chinese region, 3 other relative species (O. japonica, O. chinensis and O. intricata), and two outgroup species (Pseudomorphacris hollisi and Tetrix japonica). In the obtained sequences of O. agavisa, A% + T% was about 71.0% and 9 polymorphic nucleotide sites were observed (about 2.08%), among which no transversions were observed and only one substitution resulted in the variation of amino acid. In the case of 3 other species of Oxya, A% + T% was about 72.5% (O. chinensis) and 70% (O. japonica and O. intricata), respectively. A% + T% in the third site was much higher than the other two sites in all the analyzed Oxya species (O. agavisa, 88.3%; O. chinensis, 92.4%; O. japonica, 86.8%; O. intricata, 89.5%). Six haplotypes of O. agavisa were compared with the 3 other Oxya species and 73 polymorphic nucleotide sites were examined (about 16.9%) in all. Phylogenetic tree was constructed with the Neighbor-Joining methods using P. hollisi and T. japonica as outgroups and the confidence of nodes in the tree was evaluated by Bootstrap(1000 replicates). The NJ tree showed that different haplotypes of O. agavisa clustered together, but no clear relationships between haplotypes and the geographic regions would be inferred. The population from Mount Wuyi belongs to the peculiar group and is relatively distant to other populations. In addition, the NJ tree showed that O. agavisa was closer to O. japonica and O. chinensis closer to O. intricata, which was different from the previous results of morphologic, chromosome karyotype and bands. The relationships among different species of Oxya still need to be further clarified. PMID- 12096629 TI - [The detection of living triploid of Haliotis discus hannai--directly making chromosome sample of mantle and epipedium tentacle]. AB - Ploidy detection method of living body is a very important component part in polyploid breeding. There were few reports about chromosome ploidy detection of adult abalone (Haliotis discus hannai), except preparing adult abalone chromosome by means of chopping method using gill tissues and squashing method using gonad, also with these methods the abalone should be killed for getting these tissues. In polyploid breeding living triploid abalones are needed, so ploidy detection technique of living abalone is especially important. In this paper the orthogonal experiment of three factors and three levels L9(3(4)) using mantle and epipodium tentacle as material of living tissues which were vido immersing in PHA solution for preparing chromosome and count the frequence of cell division was first reported. Three factors and three levels were raising temperature: 16 degrees C, 18 degrees C, 20 degrees C; PHA concentration: 0.5%, 1.0%, 1.5%; sampling time: 5:00-6:00, 17:00-18:00, 23:00-24:00, respectively. The tests were repeated two times. The test results showed that the optimal patterns effecting the mitotic frequency of three factors and three levels in triploid abalone detection of chromosome from mantles and epipodium of living bodies were: raising temperature 20 degrees C, PHA concentration 1.0%, and sampling time 17:00-18:00. The order of three factors was: raising temperature-->PHA concentration-->sampling time. Under optimal pattern of three factors and three levels many clear, proper in length and well-scattered metaphase chromosomes could be obtained. The advantages of preparing chromosome using mantles and epipodium tentacle were sampling convenient, no body size restriction and no influence of survival of abalone. After using PHA treated tissues, mitotic frequency could be improved and 1.00% PHA concentration was the best. The key of chromosome preparation was the cells in mitotic metaphase, so the time of sampling was important. During 17:00-18:00, sampling of abalone many mitotic cells could be obtained because abalones were vigorously active at night. Biological zero degree of abalone was 7.6 degrees C. Under 7.6 degrees C abalone could not grow. At 20 degrees C abalones move and prey the most vigorously and grow fastest. Therefore a lot of cells are in mitotic metaphase. PMID- 12096630 TI - [Obtaining stem borer-resistant homozygous transgenic lines of Minghui 81 harboring novel cry1Ac gene via particle bombardment]. AB - A modified cry1Ac gene was generated by fusing with Lys-Asp-Glu-Lue (KDEL), an endoplasmic reticulum retention signal at the 3'-ends, with signal peptide coding sequence of Soybean kunitz trypsin inhibitor (SKTI) at the 5'-ends. Vector containing the modified cry1Ac gene coding region flanked by the corn ubiquitin 1 promoter and the nopaline synthase gene (nos) terminator with Hygromycin Phosphotransferase (hpt) gene as a plant selection marker was constructed. The modified cry1Ac gene in which toxic protein targeted to endoplasmic retention was successfully transferred into Minghui 81 (Oryza sativa L. subsp. indica), an elite restoring line of commercial CMS indica hybrid rice, through particle bombardment and obtained fertile transformants. Homozygous transgenic rice lines were obtained in the third generation exploiting self-seed set reproduction and HygromycinB selection. These transgenic lines were confirmed with polymerase chain reaction (PCR) amplification, Southern blotting and ELISA detection. Pest insect-resistant bioassay indicated that some of the homozygous cry1Ac-transgenic rice plants of T2 progeny showed high-level resistance against striped stem borer (Chilo suppressalis) at field trials. PMID- 12096631 TI - [ESTs analysis of resistance to powdery mildew in wheat at primary infected stage]. AB - A wheat line, Bai Nong 3217 x Mardler BC5F4 with resistance to powdery mildew was used in the study. A suppression subtractive hybridization cDNA library was constructed from wheat leaves challenged by Erysiphe graminis DC at primary stage. Seven hundred and sixty ESTs were acquired, and the ESTs similarity analysis based on BLASTx software was finished by comparing sequences in nr database of GenBank. Two hundred and seventy one ESTs' functions were identified in the total ESTs. The results showed that GTP-binding proteins associated signal pathway, salicylic acid pathway, MAP pathway etc were supposed to involved in the disease resistance reaction. SAR genes were rich not only in varieties but also in quantity, including five kinds of phyogenesis-related proteins, induced defence-resistance genes, heat shock proteins and genes induced by abiotic stresses etc. There are lots of evidence to testify PAL pathway, cell wall modification, cell survive system serving in the disease resistance. In the function unknown ESTs, many homologous ESTs were found from other biotic and abiotic-stresses selected cDNA libraries after BLASTn analysis, the stresses included pathogen, salt, drought, cold, high temperature etc. The novel ESTs was 16.6% in total ESTs. PMID- 12096632 TI - [Genetic analysis on restorer genes of D2-type CMS lines of common wheat]. AB - On the basis of genetic characters of D2-type CMS lines, restorer lines Yi4060, M8003, 6D/6R, GR1, 960789, Bao769-22-1 et al. were gained from crossing selection. The results of genetic analysis of F2, F1BC1 population and allelic analysis of restorer genes, successive selection of self fertility from F1 showed that fertility restoration of these restorer lines was controlled by two pair independent major-genes(dominant) and different-dosage minor genes. Authors proposed that the two pair independent major-genes should be designated as D2Rf1 D2Rf1, D2Rf2 D2Rf2. The model C2(major genes + minor genes) should be first selected in breeding restorer line. PMID- 12096633 TI - [Effects of the enhanced UV-B radiation on the body cell mitosis of the wheat]. AB - The chromosomal mutation and mitosis of the wheat (Triticum aestivum, cv. 93 4736) were observed under the condition of the enhanced ultraviolet-B radiation. The results showed that the number of cells in mitosis of the UV-B treatments was 68.09% of the control, indicating that the UV-B treatment significantly inhibited division of root tip cells in wheat. Using t-test at P < or = 0.05, there was a significant difference between the control and the treatments. The enhanced UV-B significantly increased aberration frequency of chromosomes and cells (t = 9.51, n = 3, P < 0.05). Several types of chromosomal aberration such as lagging chromosomes of chromosomes bridge, fragments, nuclear aberration occurred in both of control and UV-B treatment. However, the frequency of lagging chromosome and fragments of chromosome was increased significantly by enhanced UV-B radiation (P < 0.05). Furthermore, the UV-B treatment caused several new types of chromosomal aberrations, such as three-bundle, four-bundle and six-bundle. In our opinion, these new types of chromosomal aberrations should be called 'partition-bundle division'. Under the condition of UV-B treatment, the amount of aberrations only accounted for 2.84% of the total during the interphase. In the period, nuclear abnormality appeared mainly in forms of micronucleolus and polycaryon. In the metaphase, there mainly exist fragment chromosome, which accounted for 26.67% of the total aberrations, being secondary to lagging chromosomes. The amount of chromosomal abnormality in the anaphase and telophase was the biggest, which included the types of bridge, lagging chromosomes, etc. Among the types, the lagging chromosomes were dominant, accounting for 32.84% of the total. In this study, what was especially emphasized was the UV-B induced abnormal mitosis that was first found. In the anaphase and telophase of the treated cells, chromosomes were divided into 3 bundles, 4 bundles, or 6 bundles, respectively accounting for 8.77%, 15.43%, and 8.02% of the total abnormal cells. Three-bundle division was nonsymmetrical. The number of chromosomes in each of the three bundles was not completely equal. Four-bundle division mainly symmetrized at direction of axis, the number of each of the two bundles on the same axis was basically the same, but was different from that on another axis. Four-bundle division was conducted in two patterns of division. With respect to the much more special six-bundle division, chromosomes were regularly divided into 3 groups, symmetrizing each other. As viewed from the number of chromosomes in each bundle, it was smaller than that in normal somatic cells. In the abnormal division of cells, cell plate or cell wall between the two divided bundles was not found, leading to 'multi bundle bodies' occurrence by the end of cell division. Therefore, the abnormal division of the somatic cells should be called 'the multi-bundle division' or 'the partition bundle division'. PMID- 12096634 TI - [Genetic diversity of D-genome revealed by SSR markers in synthesized hexaploid wheat introduced from CIMMYT]. AB - Simple sequence repeat (SSR) molecular marker was used to measure the genetic diversity of D-genome in 26 synthesized hexaploid wheat (AABBDD) introduced from CIMMYT. Twenty-three D-genome specific SSR primers were selected for PCR amplification, among which 22 primers can detect polymorphism. A total of 92 alleles were identified at 23 loci using the above SSR primers, with an average of 4 alleles per locus. The 92 alleles were used to calculate Nei's similarity index (GS) and the genetic distance (GD). It was also found that the mean genetic distance between 26 synthesized hexaploid wheat was 0.4955, which was obviously high. From the above results, it can be indicated that the genetic variation of D genome in synthesized hexaploid wheat was abundant and could be used to improve the genetic diversity in wheat breeding. Interestingly, synthesized hexaploid wheat 17 and 18 shared the same D-genome donor, but three of 23 detected SSR loci were polymorphic between the two materials. Therefore, during the period of allopolyploidization, there was genetic differentiation in repeat region of donor genome. PMID- 12096635 TI - [Sequence of the ITS region of nuclear ribosomal DNA(nrDNA) in Xinjiang wild Dianthus and its phylogenetic relationship]. AB - Xinjiang is a center of distribution and differentiation of genus Dianthus in China, and has a great deal of species resources. The sequences of ITS region (including ITS-1, 5.8S rDNA and ITS-2) of nuclear ribosomal DNA from 8 species of genus Dianthus wildly distributed in Xinjiang were determined by direct sequencing of PCR products. The result showed that the size of the ITS of Dianthus is from 617 to 621 bp, and the length variation is only 4 bp. There are very high homogeneous (97.6%-99.8%) sequences between species, and about 80% homogeneous sequences between genus Dianthus and outgroup. The sequences of ITS in genus Dianthus are relatively conservative. In general, there are more conversion than transition in the variation sites among genus Dianthus. The conversion rates are relatively high, and the ratios of conversion/transition are 1.0-3.0. On the basis of phylogenetic analysis of nucleotide sequences the species of Dianthus in China would be divided into three sections. There is a distant relationship between sect. Barbulatum Williams and sect. Dianthus and between sect. Barbulatum Williams and sect. Fimbriatum Williams, and there is a close relationship between sect. Dianthus and sect. Fimbriatum Williams. From the phylogenetic tree of ITS it was found that the origin of sect. Dianthusis is earlier than that of sect. Fimbriatum Williams and sect. Barbulatum Williams. PMID- 12096636 TI - [Studies of marker screening efficiency and corresponding influencing factors in QTL composite interval mapping]. AB - Screening markers efficiently is the foundation of mapping QTLs by composite interval mapping. Main and interaction markers distinguished, besides using background control for genetic variation, could also be used to construct intervals of two-way searching for mapping QTLs with epistasis, which can save a lot of calculation time. Therefore, the efficiency of marker screening would affect power and precision of QTL mapping. A doubled haploid population with 200 individuals and 5 chromosomes was constructed, with 50 markers evenly distributed at 10 cM space. Among a total of 6 QTLs, one was placed on chromosome I, two linked on chromosome II, and the other three linked on chromosome IV. QTL setting included additive effects and epistatic effects of additive x additive, the corresponding QTL interaction effects were set if data were collected under multiple environments. The heritability was assumed to be 0.5 if no special declaration. The power of marker screening by stepwise regression, forward regression, and three methods for random effect prediction, e.g. best linear unbiased prediction (BLUP), linear unbiased prediction (LUP) and adjusted unbiased prediction (AUP), was studied and compared through 100 Monte Carlo simulations. The results indicated that the marker screening power by stepwise regression at 0.1, 0.05 and 0.01 significant level changed from 2% to 68%, the power changed from 2% to 72% by forward regression. The larger the QTL effects, the higher the marker screening power. While the power of marker screening by three random effect prediction was very low, the maximum was only 13%. That suggested that regression methods were much better than those by using the approaches of random effect prediction to identify efficient markers flanking QTLs, and forward selection method was more simple and efficient. The results of simulation study on heritability showed that heightening of both general heritability and interaction heritability of genotype x environments could enhance marker screening power, the former had a greater influence on QTLs with larger main and/or epistatic effects, while the later on QTLs with small main and/or epistatic effects. The simulation of 100 times was also conducted to study the influence of different marker number and density on marker screening power. It is indicated that the marker screening power would decrease if there were too many markers, especially with high density in a mapping population, which suggested that a mapping population with definite individuals could only hold limited markers. According to the simulation study, the reasonable number of markers should not be more than individuals. The simulation study of marker screening under multiple environments showed high total power of marker screening. In order to relieve the problem that marker screening power restricted the efficiency of QTL mapping, markers identified in multiple environments could be used to construct two search intervals. PMID- 12096637 TI - [Maximum entropy principle and population genetic equilibrium]. AB - A general mathematic model of population genetic equilibrium was constructed based on the maximum entropy principle. We proved that the maximum entropy probability distribution was equivalent to the Hardy-Weinberg equilibrium law. A population reached genetic equilibrium when the genotype entropy of the population reached the maximal possible value. In information theory, the entropy or the information content is used to measure the uncertainty of a system. In population genetics, we can use entropy to measure the uncertainty of the genotype of a population. The agreement of the maximum entropy principle and the hardy-Weinberg equilibrium law indicated that random crossing is an irreversible process, which increases the genotype entropy of the population, while inbreeding and selection decrease the genotype entropy of the population. In animal or plant breeding, we often use selection and/or inbreeding to decrease the entropy of a population, and use intercrossing to increase the entropy of the population. In this point of view, breeding is actually regulating the entropy of population. By applying the basic principle of informatics in population genetics, we revealed the biological significance of the genotype entropy and demonstrated that we can work over population genetic problems with the principles and methods of informatics and cybernetics. PMID- 12096638 TI - Overview of matrix metalloprotease modulation and growth factor protection in wound healing. Part 1. AB - Increasingly, basic research is revealing that the biochemical environment of the non-healing wound is different from that of the healing wound in ways that negatively affect multiple aspects of the healing process. These areas of biochemical difference may provide avenues of research for new therapeutic interventions. A particular biochemical finding in nonhealing wounds of diverse etiologies is the persistent elevated levels of a specific class of proteolytic enzymes known as the matrix metalloproteases or MMPs. The effects of these high levels of MMPs in the wound environment are thought to include local destruction of growth factors and their receptors as well as degradation of granulation tissue components. MMPs are fast gaining attention as potential causative factors in impaired healing, and recently, strategies have been focused on preventing or reducing their levels, once elevated, in the wound environment. A proprietary combination of bovine collagen and oxidized regenerated cellulose has demonstrated an ability to not only reduce elevated wound proteases but also sequester and protect wound growth factors and promote cellular chemotaxis and proliferation. PMID- 12096640 TI - Recent advances in tissue engineering: an invited review. AB - Tissue formation within the body, as part of a development or repair process, is a complex event in which cell populations self-assemble into functional units. There is intense academic, medical, and commercial interest in finding methods of replicating these events outside the body. This interest has accelerated with the demonstration of the engineering of skin and cartilage tissue in the laboratory and there is now worldwide activity in the in vitro regeneration of tissues including nerve, liver, bone, heart valves, blood vessels, bladder, and kidney. Approaches to tissue engineering center on the need to provide signals to cell populations to promote cell proliferation and differentiation. This review considers recent advances in methods of providing these signals to cells using examples of progress in the engineering of complex tissues. PMID- 12096639 TI - The role of oxidized regenerated cellulose/collagen in chronic wound repair. Part 2. AB - Normal wound healing is a carefully controlled balance of new tissue formation and destructive processes necessary to remove damaged tissue. Within this complex environment many points of regulation precisely control the biological processes necessary to achieve normal wound repair. An alteration in any of these physiological processes can lead to the formation of a chronic wound. In general, chronic wounds are characterized by a prolonged inflammatory phase, which ultimately results in elevated protease activity and the subsequent degradation of growth factors and other positive wound healing factors; the overall effect is impaired healing. PMID- 12096641 TI - A long-term clinical study on dislocated ankle fractures fixed with self reinforced polylevolactide (SR-PLLA) implants. AB - Sixteen patients with dislocated ankle fractures fixed between 1988 and 1991 with self-reinforced poly(L-lactide; SR-PLLA) screws and/or rods were followed up after 8.6 to 11.7 years (mean 9.6 years) at the Department of Orthopaedics and Traumatology, Helsinki University Central Hospital. In all patients accurate reduction of the fractures was retained and uneventful bony union was achieved. Good or excellent long-term functional results were observed in 15 out of 16 patients. One patient had post-traumatic osteoarthritis. In 5 patients, a late tissue reaction was observed over an extruding screw head with mild symptoms, which led to removal of small palpable masses. There were two superficial wound infections, one after a primary operation and one caused by a late tissue reaction after an operation. The correct operative technique, where all extruding extraosseous SR-PLLA material should be removed during the primary operation, should be followed. PMID- 12096642 TI - Frictional torque and wear of retrieved hip prostheses: a comparison between alumina/PE and Co-Cr/PE prostheses. AB - Frictional torque and wear were measured in 67 retrieved cups with internal diameters of 32 mm. Thirty cups were combined with alumina heads and 37 were combined with metal heads. A lower prevalence of cup loosening was observed in alumina-polyethylene implants than in metal-polyethylene implants. No relation between frictional torque and stability of implants was detected for either type, whereas retrieved cups of alumina-polyethylene implants showed lower frictional torque than those of metal-polyethylene implants. Less wear was observed in the cups of alumina-polyethylene implants (0.13 mm/y) than in those of metal polyethylene implants (0.19 mm/y). Less wear was observed in cups without loosening than in those with loosening in both types. These results suggest that wear has a greater influence on the stability of cups than frictional torque. PMID- 12096643 TI - A tribute to Richard F. Edlich, M.D., Ph.D., editor-in-chief. Journal of long term effects of medical implants. PMID- 12096644 TI - Nurse-training at the 'Dreadnought' Hospital, Greenwich in the late 19th century: Part I. AB - By the early 20th century, nurse-training at the Seamen's Hospital Society 'flagship' hospital at Greenwich was at least as well established as that in the London teaching hospitals. It had its origins in the late 19th century--in the light of Florence Nightingale's nursing reforms (she had initiated the training school at St Thomas's in 1860). Although Nightingale's major interests lay in the welfare of the soldier, she was clearly concerned with the health of the seafarer also. However, those early days of nurse training at the Dreadnought hospital posed numerous problems, not only for nurse training but for nursing in general. The second part of this article will appear in the next issue of the International History of Nursing Journal. PMID- 12096645 TI - The contribution of the Hospital Saturday Fund to nursing history: evaluation of a primary source. AB - The Hospital Saturday Fund was established as a means of providing much-needed contributions to the voluntary hospitals. Subscribing to the Fund ensured the contributor's care, but, additionally, they could sponsor a 'deserving object'. The annual reports of the Fund provide a number of insights in relation to the social, economic and political history of Liverpool. The reports also provide some insights into nursing in the city during the period 1888-1927. Following documentary analysis, the paper discusses some of these findings, namely district nursing, services for women and convalescence, and evaluates how useful this source may be to nursing history. PMID- 12096646 TI - The use of prolonged baths in psychiatric care: understanding through fiction. AB - Two autobiographical novels containing descriptions of prolonged baths from the patient's perspective are referred to and analysed. They include depictions of patients who are treated roughly. Staff do not interact verbally with the patients, who attempt to discern the intentions and moods of the staff through their body language. The depictions from the patient's perspective are discussed in relation to nurses' narratives with the use of medico-historical references and psychological literature. PMID- 12096647 TI - Looking back to move forward: cadet nursing in the 1970s. AB - Linda Witchell recalls her experiences as a cadet nurse (1972-1974), highlighting the values and pitfalls encountered and suggesting lessons that could be learned. PMID- 12096648 TI - Health care for all children: the beginnings of school nursing 1904-1908. AB - This paper explores the origins of school nursing in the school medical service. The work and employment of school nurses is discussed, presenting some interesting resonances with today's school nursing practice. It is regrettable that there is no single, clear voice from a school nurse, but other sources were found, and the humanity added by nurses to the screening and treatment of children is apparent. A second paper, to be published in the next issue of the International History of Nursing Journal, considers the work of school nurses in the period immediately after World War II. PMID- 12096649 TI - [Progress in diagnostic echocardiography of the heart]. PMID- 12096650 TI - [Contrast echocardiography: clinical applications and future prospects]. AB - BACKGROUND: Contrast echocardiography has been used as a clinical method for more than 20 years. Using conventional ultrasound techniques the clinical use of contrast was limited. Now the development of "contrast specific" imaging modalities has increased the indications for contrast echocardiography. CONTRAST AGENTS: For clinical use two classes of contrast agents are available: 1. "right heart" contrast media (Echovist, agitated solutions), which do not cross the pulmonary vascular bed following intravenous injection and which can be used with conventional (fundamental) imaging methods, 2. "left heart" contrast media (Levovist, Optison, SonoVue) which need "contrast specific" imaging modalities for optimal use. INDICATIONS: Despite of the developments in Doppler methods and transesophageal echocardiography "right heart" contrast media still are needed in some patients with atrial and pulmonary shunts, complex congenital heart disease, noisy Doppler recordings of tricuspid regurgitation. For "left heart" contrast media improvement of endocardial border definition is the most important indication, which has been validated in a series of well performed studies. Therefore contrast enhanced recordings are recommended in the clinical echo laboratory when unenhanced recordings are suboptimal. Coronary flow reserve of the LAD can be measured using contrast enhanced Doppler echocardiography. All contrast specific imaging modalities provide assessment of myocardial perfusion. The previously used imaging modalities (Harmonic B mode, Pulse Inversion and Harmonic Power Doppler) did not provide sufficient myocardial contrast signals using real-time imaging. Although intermittent imaging resulted in good myocardial opacification, this modality did not gain wider clinical acceptance. NEW TECHNOLOGIES: Using new contrast specific imaging technologies like Power Pulse Inversion, Power Modulation and Coherent Imaging myocardial perfusion can be evaluated in "real-time". Thus simultaneous assessment of left ventricular wall motion and myocardial perfusion became a reality and facilitated the data acquisition. New ultrasound contrast media like SonoVue can be used for all imaging modalities. Recent studies have demonstrated that the information derived from myocardial contrast echocardiography provides clinically relevant information on top of the findings obtained from conventional left ventricular wall motion analysis. PMID- 12096651 TI - Myocardial perfusion imaging using contrast echocardiography. AB - BACKGROUND: Intense work during the last two decades has brought forth the use of myocardial contrast echocardiography to the clinical threshold for the diagnosis and evaluation of coronary artery disease. CLINICAL USE: A number of ultrasound contrast agents have been developed that act as red blood cell tracers and display myocardial perfusion when imaged by dedicated ultrasound imaging modalities. A considerable amount of experimental and clinical research has shown that myocardial contrast echocardiography can aid in the recognition of acute and chronic myocardial infarction, viable myocardium, and functionally significant coronary stenoses. Comparison of this technique to nuclear imaging and coronary arteriography has demonstrated excellent diagnostic accuracy in the evaluation of various coronary syndromes. Optimal practice of perfusion imaging requires a thorough knowledge of microbubble characteristics and imaging modalities, as well as good experience in the method. PERSPECTIVES: The technique continues to evolve from intermittent gated examination to real-time perfusion imaging that allows evaluation of both perfusion and functional parameters. The opportunity to target sites of pathology with specially engineered microbubbles could also aid in many therapeutic applications besides diagnostic imaging. PMID- 12096652 TI - [3-D echocardiography: new developments and future prospects]. AB - Due to limitations in transthoracic and occasionally transesophageal 2-D echocardiography with respect to volumetric analysis and morphologic and functional assessment in patients with congenital malformations and valvular heart disease, additional diagnostic tools have been established. In parallel with the rapid evolution in computer technology, 3-D echocardiography has grown into a well-developed technique, such as volume-rendered 3-D reconstruction, capable of displaying dynamic morphology depicting depth of the structures, their attachment, and spatial relation to the surrounding tissue. Nevertheless, the complexity of data acquisition and data processing required for adequate dynamic 3-D echocardiographic imaging and volumetric analysis does not allow to use this approach routinely. The commonly used dynamic 3-D echocardiography means off-line computer-assisted image reconstruction from a series of cross-sectional echocardiographic images using currently available transesophageal and transthoracic transducers. Alternatively, real-time 3-D echocardiography based on novel matrix, phased-array transducer technology has been introduced. Although this technique can be easily combined with any routine examination, its clinical use is limited because of a lower image quality in comparison with dynamic 3-D echocardiography. Up to now, there is no transesophageal approach available using real-time 3-D echocardiography. Recently, dynamic 3-D echocardiographic technique has matured noticeably. Beside the well-known sequential scanning, which is characterized by a fixed probe and patient in space and predetermined motion of the transducer, the freehand scanning using an electromagnetic location system has found its way to clinical environment. The main advantage of this technique is that the transducer can be freely moved by the examiner and, thus, the data set acquired within a routine examination. Also 3-D rendering and display have been developed further. In this respect, especially the "real-time rendering mode" allowing the reconstructed 3-D image to be animated and moved in space and to look at it from different perspectives has gained increasing acceptance. In valvular heart disease, reconstructive surgical treatment is aspired. 3-D echocardiographic imaging is the only technique providing "surgical views" prior to opening the heart. It is capable of distinguishing particular destructive substructures of the valves and the valvular apparatus. Especially in mitral valvular reconstruction, it is of clinical importance to achieve optimal surgical results. With respect to volumetric and mass analysis, 3-D echocardiography is more accurate and reproducible in comparison with conventional 2-D analysis. It provides data independent of geometric assumptions, what may considerably influence the results in the presence of wall motion abnormalities, especially in aneurysmatic ventricles. Volumetric analysis of the aneurysmal portion may also be helpful prior to surgical resection. 3-D echocardiography can also be recommended as a valuable additional approach to atrial septal defect (ASD), corrected transposition of the great arteries, cor triatriatum, and, within limits, to ventricular septal defect (VSD) as well. Especially with respect to ASD and VSD, the potential significance of 3-D echocardiography prior to device closure is emphasized. At present, its additional information in decision-making and the increasing number of clinical cases that can be addressed and answered already justify the clinical use of this technique. PMID- 12096654 TI - New imaging techniques in the assessment of patients with aortic stenosis. AB - BACKGROUND: Currently, conventional Doppler echocardiography is the noninvasive diagnostic technique to be used in the evaluation of the vast majority of patients with aortic stenosis. However, several more recent imaging techniques are being progressively introduced in the cardiologist's diagnostic armamentarium. Some of these techniques may offer additional information that could be of help in some patients with aortic stenosis. TECHNIQUES: Electron beam computed tomography may evaluate and monitor the degree of aortic valve calcification. Three-dimensional echocardiography and magnetic resonance imaging may be of help in the measurement of the aortic valve area in cases with suboptimal acoustic window. The degree of left ventricular involvement may be accurately assessed and monitored by magnetic resonance imaging, electron beam computed tomography, tissue Doppler and automated segmental motion analysis. Finally, abnormalities in left ventricular metabolism may be evaluated by positron emission tomography and magnetic resonance imaging. CONCLUSION: Some of these techniques could be applicable in the future in the evaluation of patients with aortic stenosis. PMID- 12096653 TI - Real-time three-dimensional echocardiography for improved evaluation of diastolic function using volume-time curves. AB - BACKGROUND: Accurate assessment of left ventricular function is of the greatest importance in clinical cardiology for decision making. Diastolic dysfunction is getting more concern as a cause of heart failure while, currently used non invasive modalities for diagnosing diastolic abnormalities have significant limitations. Dynamic left ventricular volume change was applied for the evaluation of diastolic function by various techniques that have been demonstrated to be of diagnostic value. However, it has not been accepted into clinical practice because existing techniques are either invasive, inaccurate, expensive or time consuming. REAL-TIME THREE-DIMENSIONAL ECHOCARDIOGRAPHY: Real time three-dimensional (3-D) echocardiography is a new ultrasound technique that provides transthoracic volumetric images of the heart in real time. Thereby, the acquired images are ideally suited for the assessment of dynamic left ventricular volume change. Generation and analysis of left ventricular volume-time curves by real-time 3-D echocardiography has been demonstrated to be feasible in normal subjects and patients and accuracy of volume-time curves was good compared to magnetic resonance imaging. We compare the new real-time 3-D echo approach with the advantages and limitations of existing noninvasive and invasive techniques. PMID- 12096655 TI - [Echocardiography flow quantification for determining the severity of heart valve insufficiency]. AB - BACKGROUND: Beyond conventional echocardiographic Doppler methods allowing only semiquantitative estimation of the severity of valvular regurgitation, new approaches are attempting to quantify regurgitant flow as a measure of left ventricular volume overload. Different concepts are leading to significant differences in accuracy and feasibility in clinical routine between the methods. We are reviewing the existing methods with their advantages and limitations as well as the underlying hemodynamic concepts. ECHOCARDIOGRAPHIC METHODS FOR ESTIMATION OF SEVERITY: Semiquantitative methods are jet area/jet length method, proximal jet width as well as the pressure half-time method. Determination of regurgitant flow is permitted by means of the continuity method, PISA ("Proximal Isovelocity Surface Area") method, ACOM ("Automated Cardiac Output Measurement") technique, and the PVI ("Power-Velocity Integral") method. Grading of severity is usually based on a scale from 1 to 3 or 1 to 4 where semiquantitative methods are limited by a significant overlap of the individual degrees and therefore only provide rough estimates of severity. Compared to this, quantitative methods allow quantitative determination of regurgitant flow, regurgitant volume, regurgitant fraction, and effective regurgitant orifice area based on different Doppler flow measurements, that are again affected by specific limitations: (1) the continuity method is considerably limited because it is measuring the flow through two different valves; (2) the PISA method is dependent on the geometry of the proximal flow convergence zone and requires highly skilled observers; (3) the ACOM method is also dependent on the geometry of the proximal flow convergence zone und ideally requires three-dimensional color Doppler datasets; (4) the PVI method provides direct measurement of regurgitant flow from a pulsed Doppler signal of sufficient high quality, however, this method is not widely available, yet. For the decision which method to apply the following should be taken into account: (1) interpretation of semiquantitative findings depends on the extent by which the methods' estimation of flow deviates from a measurement of flow; (2) accuracy of the quantitative methods depends on the underlying hemodynamic concept and the simplifying assumptions. CONCLUSION: Current echocardiography provides a broad spectrum of semiquantitative und quantitative Doppler methods for the estimation of regurgitant flow. Since semiquantitative methods only allow rough estimates of severity, quantitative measurement of regurgitant volume/fraction or effective regurgitant orifice area should be attempted in any case with uncertainty whether regurgitation is mild, moderate, or severe. PMID- 12096656 TI - Therapeutic ultrasound in cardiology. AB - Ultrasound can be exploited to derive therapeutic results by using its bioeffects such as creation of mechanical vibrations, localized cavitations, microstream formation, physicochemical changes and thermal energy. Extensive in vitro and animal investigations during the last 2 decades have laid a foundation for ultrasound energy to be used for treatment purposes in various medical specialties. In the area of cardiovascular diseases, ultrasound could be used for thrombolysis, adjunct to coronary interventions, drug delivery, local gene transfer, and creating therapeutic lesions. The dispensation approaches to therapeutic ultrasound are varied, from the use of low- to medium-range frequency, low to focused high intensity, and catheter-based to external devices. Catheter-based ultrasound could be useful for intracoronary thrombolysis, and external ultrasound instrument with transcutaneous delivery could be of use in applications such as creation of myocardial lesions, peripheral vessel thrombolysis, and drug and gene delivery. Adjunct administration of microbubbles has been found to enhance thrombolysis, and drug and gene therapy. Ongoing studies strongly suggest that therapeutic ultrasound could have an important role in cardiovascular disorders associated with thrombosis, inflammation, atherosclerotic disease, and arrhythmias. PMID- 12096657 TI - [Current aspects of pathophysiology and therapy of pulmonary hypertension (PHT)]. PMID- 12096658 TI - Setting priorities in waiting lists. PMID- 12096659 TI - Brain imaging in psychiatry: what has it done for the patient? PMID- 12096660 TI - Neuroimaging in schizophrenia: from theory to practice. AB - Schizophrenia is a serious mental illness that affects 1% of the population. It is beset with problems of diagnosis and difficulties in assessing treatment. Neuroimaging has long contributed to our understanding of schizophrenia but recent developments suggest it may ultimately play a practical role in its early detection and management. PMID- 12096661 TI - Brain imaging: a key to understanding depression. AB - According to the World Development Report 1993 one third of people with a mental illness suffer from a depressive disorder. There is a need for early diagnosis and effective treatment in order to limit the impact on patients' lives. Knowledge gained from brain imaging research may help to improve our understanding and treatment of depression. PMID- 12096662 TI - What neuroimaging tells us about psychopathic disorders. AB - The concept of psychopathy and its neurobiological basis are discussed. Structural (computed tomography, magnetic resonance imaging) and functional (positron emission tomography, functional magnetic resonance imaging, single photon emission tomography) studies in antisocial and psychopathic samples are reviewed in this article. PMID- 12096663 TI - Neuroimaging to predict preclinical Alzheimer's disease. AB - Alzheimer's disease is common in the elderly and causes tremendous distress to patients and their carers. With the advent of newer pharmacological treatments, significant improvement in the early diagnosis of Alzheimer's disease is required. This article examines the usefulness of neuroimaging techniques to predict Alzheimer's disease in prediagnosis individuals. PMID- 12096664 TI - Agalsidase alfa: specific treatment for Fabry disease. AB - Fabry disease is a rare genetic lysosomal storage disorder characterized by a deficiency of the enzyme alpha-galactosidase A. The recent availability of enzyme replacement therapy with agalsidase alfa offers specific treatment for this serious, progressive condition. PMID- 12096665 TI - Diagnosis and immediate care of injuries to the elbow and forearm. PMID- 12096666 TI - Intestinal ischaemia. AB - Over the past three decades major advances have been made in the diagnosis and management of mesenteric ischaemia. Early identification and treatment of this condition has reduced its mortality and morbidity. This review discusses the causes, preliminary and definitive investigations, and treatment protocols relating to gut ischaemia. PMID- 12096667 TI - Using a competence framework to select future medical specialists. AB - Postgraduate deans in one UK region have developed a competence framework for selection of specialist registrars. This paper describes the process by which this was developed and adapted to 62 specialties. Evaluation demonstrated high levels of acceptability for applicants and to a lesser extent for selectors. An external review confirmed that the process was in keeping with good employment practice. PMID- 12096668 TI - Alcoholic hepatitis with ascites may mimic advanced ovarian malignancy. PMID- 12096670 TI - The lesson from a yellow psychotic patient. PMID- 12096671 TI - Acute leukaemia masquerading as acute abdomen. PMID- 12096672 TI - Complications of catheterization. PMID- 12096673 TI - Obtaining informed consent. PMID- 12096675 TI - Acute renal failure in the intensive care unit: does it really increase mortality? PMID- 12096674 TI - Counselling patients before rectal examination. PMID- 12096676 TI - The role of traditional mediterranean type of diet and lifestyle, in the development of acute coronary syndromes: preliminary results from CARDIO 2000 study. AB - The traditional Mediterranean type of diet is inversely associated with the risk of developing coronary heart disease. Aim of this study is to investigate the relationships between Mediterranean type of diet, various lifestyle factors and their contribution to the risk of developing acute coronary syndromes. During 2000, a case-control study was conducted (CARDIO2000) and food items as well as olive oil consumption were recorded in addition to various cardiovascular risk factors. Based on the estimated risk model we found that the Mediterranean type of diet reduces significantly the risk of developing acute coronary syndromes even in the presence of unfavorable lifestyle situations, such as sedentary life, smoking habit, as well as hypertension, hypercholesterolemia and diabetes mellitus. PMID- 12096677 TI - Pathophysiology of vibration-induced white fingers--current opinion: a review. AB - Vibration-induced white finger (VWF) is secondary Raynaud's phenomenon caused by vibrating hand tools. Initially described in 1911 by Loriga, the pathophysiology of VWF remains unclear. This review attempts to summarize the current common and the most modern theories of the pathogenesis of VWF and propose a model of multifactorial etiology. The role of vasoregulatory abnormalities--autonomic dysfunction, local vasoregulatory abnormalities, of the endothelial cell control, of hemostasis, fibrinolysis and hemorrheology, of the leukocyte activation and the levels cell adhesion molecules and pro-inflammatory cytokines have been reviewed. PMID- 12096678 TI - Prevalence of rubella-specific IgG antibody in non-immunized pregnant women in Maiduguri, north eastern Nigeria. AB - The prevalence of IgG antibody specific to the rubella virus was assessed in 207 consenting pregnant women. The women were in the age range of 14 to 40 years (mean 25.8 years) and parity range of 0 to 8 (mean para 4), in their first and second trimesters of pregnancy. All attended the antenatal clinic of a tertiary health facility (University of Maiduguri Teaching Hospital) in northeastern Nigeria. Of the 207 serum samples tested, 112 (54.1%) were positive for rubella IgG antibody. In this area therefore, the proportion of susceptible pregnant women to rubella virus is up to 46%. The results from this study also indicated that with increased maternal age the percentage of immune women increased significantly (P = 0.04382) from 23.8% in the 14 to 19 years age group to 74.4% in the age group of 30 to 40 years. There was again a gradual increase in rubella seroprevalence from 43% amongst primigravidae to 59% and 78% in multiparous and grandmultiparous women respectively. The pregnancy outcome was normal in 27% of the women studied, with miscarriage occurring in 24.6% and 24.2% each had a premature delivery and stillbirth. There were 30 (58.8%) miscarriages, 27 (54%) premature deliveries and 30 (60%) stillbirths, with no clinically detectable malformations in 25 (44.6%) of all the deliveries from the immune women. There is therefore, an increase in the number of rubella immune women with each of the pregnancy outcomes compared to the non-iminune ones. Hence, the need to protect children of these susceptible women from contracting congenital rubella and its sequelae by including routine rubella vaccination of all women of childbearing age in the current programme on immunization. PMID- 12096679 TI - The effect of low level radiofrequency electromagnetic radiation on the excretion rates of stress hormones in operators during 24-hour shifts. AB - The aim of the study was to investigate the effect of long term exposure to low level radiofrequency (RF) electromagnetic (EM) radiation on the excretion rates of stress hormones in satellite station operators during 24-hour shifts. Twelve male operators at a satellite station for TV communications and space research were studied during 24-hour shifts. Dosimetric evaluation of the exposure was carried out and showed low level exposure with specific absorption of 0.1127 J.kg 1. A control group of 12 unexposed male operators with similar job task and the same shift system were studied, too. The 11-oxycorticosteroids (11-OCS), adrenaline and noradrenaline were followed by spectrofluorimetric methods on 3 hour intervals during the 24-hour shifts. The data were analyzed by tests for interindividual analysis, Cosinor analysis and analysis of variance (ANOVA). Significant increase in the 24-hour excretion of 11-OCS and disorders in its circadian rhythm, manifested by increase in the mesor, decrease in the amplitude and shift in the acrophase were found in the exposed operators. The changes in the excretion rates of the catecholamines were significant and showed greater variability of both variables. The long term effect of the exposure to low-level RF EM radiation evoked pronounced stress reaction with changes in the circadian rhythm of 11-OCS and increased variability of catecholamines secretion. The possible health hazards associated with observed alteration in the stress system need to be clarified by identification of their significance and prognostic relevance. PMID- 12096680 TI - Smokers--vitamin C--hypercholesterolaemia. AB - Cigarette smoking is one of the major public health problems of our times. Smokers are damaged by free radicals formed during smoking. The antioxidant capacity of smokers is reduced. Vitamin C (ascorbic acid) deficiency in heavy smokers with a total cholesterol more than 6.7 mmol/l ranges from 35-55%. It is mediated by smoking, by its low dietary intake by preferential high dietary intake of saturated and unsaturated fats. Vitamin C is an important water soluble antioxidant, therefore its intake can at least partially reduce the unfavourable effect of smoking caused by the attack of free radicals on the organism when smokers are unable to quit smoking. PMID- 12096681 TI - Determination of normal concentration levels of Cd, Pb, Hg, Cu, Zn and Se in urine of the population in the Czech Republic. AB - Knowledge of normal levels of concentrations of trace elements (Cd, Pb, Hg, Cu, Zn, and Se) in the population serves, among others, in design of regulations concerning health protection, determination of exposition limits and prevention of diseases caused by deficiency of trace elements. Concentrations of the named elements in urine of the Czech population were determined by means of atomic absorption spectrometry. The urine has been collected during 1996-2000 from 1192 individuals (816 males and 376 females, average age 34.6 years) and 2008 children (1052 boys and 956 girls, average age 9.9 years). Mineralization in a microwave digestion system was used in sample preparation. The accuracy of results was checked by means of the Control Materials Seronorm 403,125 and BioRad 69,041. Values of concentrations of the trace elements in urine found for adult (medians) were 0.36 microgram Cd.g-1 creatinine, 10.6 micrograms Cu.g-1 creatinine, 0.68 microgram Hg.g-1 creatinine, 3.3 micrograms Pb.g-1 creatinine, 6.2 micrograms Se.g-1 creatinine, and 397 micrograms Zn.g-1 creatinine, respectively. Statistically significant differences between men and women have been found in the concentrations of Cu and Hg. In the juvenile population following concentrations have been found: 0.29 microgram Cd.g-1 creatinine, 16.1 micrograms Cu.g-1 creatinine, 0.32 microgram Hg.g-1 creatinine, 4.8 micrograms Pb.g-1 creatinine, 10.2 micrograms Se.g-1 creatinine and 460 micrograms Zn.g-1 creatinine. Statistically significant differences between boys and girls were found only in Cu and Hg concentrations. Concentrations of the studied elements correspond to the published values concerning population not exposed professionally. PMID- 12096682 TI - The "cost" of silence. AB - Changes in the organism of control desk operators caused by difficulties faced during productionally relevant speech information processing under conditions of intense occupational noise, represent the psychophysiological "cost" of noise masking. Its presence attests to difficulties in processing before deterioration of the quality of the latter while its quantitative characterisation correlates positively with the degree of the masking effect of noise and with the neuropsychic stress of this particular type of work. For this reason an attempt is made to demonstrate noise masking under laboratory conditions with it approximate quantitative evaluation at two signal/noise ratios- +15 and The statistically significant differences between the values of pulse rate, plethysmography and vocal exertion at the above ratios point to both its presence, and to the necessity to be considered when assessing the occupational burden and standardisation of noise intensity in productions associated with speech signals. PMID- 12096683 TI - Genetic-biochemical criteria for individual sensitivity in development of occupational bronchopulmonary diseases. AB - Human individual sensitivity to health-hazardous occupational factors and probability of developing chronic lung diseases depend on genetic variation of serum and erythrocytic proteins. The present work was aimed at studying the phenotypes of serum and erythrocytic proteins in patients with occupational respiratory diseases. We studied 7 highly polymorphic genetic systems the varieties of which may be connected with development of bronchopulmonary pathology (BPP) and the immune status of the body: proteinase inhibitor (Pi), third component of the complement (C3), transferrin (Tf), group-specific component of blood serum (Gc), haptoglobin (Hp), erythrocytic glyoxalase (Glo) and phosphoglucomutase (PGM) in patients with chronic bronchitis, silicosis, occupational bronchial asthma and in the control group consisting of Moscow population not exposed to occupational hazards and apparently healthy workers of an engineering plant. Considerable differences were revealed in genetic structure of the patients with bronchopulmonary pathology as compared with the apparently healthy people along a series of Integrated system: proteinase inhibitor (Pi), C3, Tf, Gc, PGM. Comparison of the study groups by significant differences in the aggregate of the genetic information obtained suggests that 5 (HP, C3, Tf, Pl, PGM1) of the 7 studied systems showed the hereditary features of silicosis. The gene carriers Hp*2, C3*F, PGM1*2-, TF*D, GC*R due to peculiar biochemical processes appear to have less adaptive potentialities and a greater likelihood of the disease on exposure to industrial factors. The examined patients with chronic bronchitis showed an increase in the variant of GC*2 and of a rare variants of proteins GC*R and Pi*S, the patients with occupational bronchial asthma showed an increase in the variant of Hp*2 and of a rare variant Pi*S. Such studies could be useful for assessment and forecast of individual risk of occupational diseases. PMID- 12096684 TI - Virus isolation data from water of some European rivers: an overview. AB - The present overview is the result of our scrutiny of data concerning the presence of viruses in the water of diverse European rivers. These data were assembled from the published literature--articles, doctoral theses and reports from investigations conducted by environmental virologists beginning during the final years of the sixth decenium of the twentieth century, first in France, Czechoslovakia, Romania, Ukraine, U.S.A. then in other European and countries worldwide. The overview covers the methodology referring both to water sampling (by gauze pads or by grab procedures), and to virus detection methods, including virus concentration from large volumes of water and the inoculation of virus concentrates on cell cultures and/or into suckling mice. Other more recently elaborated methods of virus detection and identification, consisting of immunological tests, as the enzyme immunoassay (EIA), immunofluorescence (IF), or the genetic techniques, as the molecular hybridization and flow cytometry (FC), were also applied. The obtained results refer to the virus positivity with the specification of virus types and various virus contamination levels of these waters. Finally, the ways by which the viral contamination of the searched river waters might be demonstrated and, ways by which the human body can be contaminated by the virus polluted river waters are discussed, and some conclusions and recommendations are formulated. PMID- 12096685 TI - Comparative investigation of airborne culturable microorganisms in sewage treatment plants. AB - The present study investigated emissions and emmissions of airborne microorganisms (mesophilic bacteria, Escherichia coli, molds, Aspergillus fumigatus, thermophilic actinomycetes/bacilli) in sewage treatment plants. For the aerobiological investigations three sewage treatment facilities with an activated-sludge process, capacities between 2000 and 28,000 PE and different cleaning steps were selected. The measurements of microorganism emission were conducted in the area of the intake (screen), in the area of biological treatment (activated sludge tank) and at a distance of 10 m from the activated sludge tanks. In order to determine the emmission, additional measurements were conducted leeward of the plant at a distance of 200 m. Samples were taken using four parallel six-stage Andersen 1 AFCM volumetric samplers. In the area of the intake counts for bacteria were 7.4 x 10(2) CFU/m3 (median), for thermophilic actinomycetes 1.8 x 10(1) CFU/m3, for thermophilic bacilli 7.1 x 10(1) CFU/m3, for molds 2.4 x 10(3) CFU/m3 and for Aspergillus fumigatus 1.8 x 10(1) CFU/m3. Only isolated airborne coliform recoveries, i.e. E. coli, were detected. In the area of the activated sludge tank, in the adjoining area (10 m) and in the vicinity of the plants (200 m), the counts for all microorganism groups investigated corresponded to natural conditions. The results show that the counts of culturable aerogenic microorganisms in and in the immediate surrounding of the sewage plants investigated are low. Although the possibility of an infection through inhalation cannot be ruled out, the direct contact with sewage is much more critical. PMID- 12096686 TI - MLEE and PFGE characterization of Neisseria meningitidis serogroup C and B isolated in the Slovak Republic in 1998. AB - In the Slovak Republic the incidence and mortality of invasive meningococcal disease increased after 1995 when the new meningococcal clone of Neisseria meningitidis C:2a:P1.2,P1.5, ET-1.5/37 emerged. The new clone spread between 1995 and 1998 throughout the whole country. Morbidity of invasive meningococcal disease was 1.6/100,000 of the population and fatality reached the highest level of 23% in the Slovak Republic in 1998. The new clone caused a new emergent epidemiological and clinical situation. The occurrence of invasive meningococcal disease caused by this clone has continually risen since 1995. In 1998 72% of all diseases in Slovakia were caused by serogroup C. The emerging clone C:2a:Pl.2,P1.5 represented 74% of the serogroup C isolates. Clonality and genetic diversity of 15 selected meningococcal strains causing invasive meningococcal disease was compared by multilocus enzyme electrophoresis (MLEE) and DNA macrorestriction analysis by pulsed-field gel electrophoresis (PFGE). The strains of serogroup C and B were isolated in all regions of Slovakia in 1998. The majority of isolates belong to hypervirulent clone ET-15 as determined by MLEE. By PFGE a higher degree of diversity was observed. PMID- 12096687 TI - The comparison of aluminium effects and its uptake by Escherichia coli in different media. AB - The investigation of the toxic effects and the uptake of aluminium by Escherichia coli in growth medium (GM) and in physiological solution (PS) have been studied. The toxicity was quantitatively determined according to the decrease of the colony forming units (CFU) in the physiological solution, that is its growth inhibition in the growth medium, vs. the aluminium concentration and incubation time, at pH 5.2, 6.2 and 7.2. The uptake of aluminium was investigated by determining the intracellular aluminium in dry weights (DW), by graphite fumace atomic absorption spectrophotometry, considering that aluminium adsorbed to the cell surface was removed by washing with EDTA solution. The results show that toxicity and accumulation increase with the increase of the aluminium concentration and incubation time. However, the linearity of these functions was lost at higher values, which indicate dependence on time and concentration saturation. The effect of pH was specific, and correlated with the form of aluminium in solution. The increase in toxicity as the pH decreases, suggests that the Al(H2O)6(3+) ion is the major toxic form, among the remaining present ones aluminium in aqueous media. The results also show that the aluminium in a concentration range from 0.10 to 10.0 mg/l toxic to E. coli in PS, was significantly less toxic for bacteria in the GM, mainly because of living conditions and the accessibility of free Al. PMID- 12096688 TI - It's not hard--it's just CPD (Continuing Professional Development). PMID- 12096689 TI - Developing high quality low cost healthcare goods. AB - The quality and cost of healthcare is a major concern in both Britain and America. Yet, despite much debate and many initiatives, the provision of healthcare is often unreliable and expensive (Goldsmith, 2001). In this article by Dr Stephen Fox, it is proposed that wider application of design methodologies during the development of healthcare goods could make a significant contribution to increasing quality and reducing costs. PMID- 12096691 TI - Airmec launches HazMat services. PMID- 12096690 TI - Special database assists choice of materials. PMID- 12096692 TI - Knowledge pooling on equipment fault diagnosis. PMID- 12096693 TI - Clinical engineering service benefits from ongoing challenge. PMID- 12096694 TI - The journey: from science via engineering through to technology. PMID- 12096695 TI - Understanding social factors and inequalities in health: 20th century progress and 21st century prospects. AB - The development of social epidemiology and medical sociology over the last half of the 20th century, in which Leo Reeder played a central role, transformed scientific and popular understanding of the nature and causes of physical health and illness. Viewed in the early 1950s as shaped almost entirely by biological processes and medical care, physical health and illness are now understood to be as much or more a function of social, psychological, and behavioral factors. Utilizing a stress and adaptation conceptual framework, social epidemiology has identified a broad range of psychosocial risk factors for health, most notably: (1) social relationships and support; (2) acute or event-based stress; (3) chronic stress in work and life; and (4) psychological dispositions such as anger/hostility, lack of self-efficacy/control, and negative affect/hopelessness/pessimism, with new risk factors continuing to be identified. However, proliferation of risk factors must be balanced by conceptual integration and causal understanding of the relationships among them, their causes, and consequences. One source of such integration and understanding has been the rediscovery of large and persistent socioeconomic and racial-ethnic disparities in health. Socioeconomic position and race/ethnicity shape individuals' exposure to and experience of virtually all known psychosocial, and well as many environmental and biomedical, risk factors, and these risk factors help to explain the size and persistence of social disparities in health. Improving the socioeconomic position of a broad range of disadvantaged socioeconomic and racial ethnic strata constitutes a major avenue for reducing exposure to and experience of deleterious risk factors for health, and hence for improving the health of these groups and the overall population. This in turn requires better understanding of the macrosocial forces that influence the socioeconomic position of individuals. PMID- 12096696 TI - Outcomes in the sociology of mental health and illness: where have we been and where are we going? AB - Sociologists of mental health and illness have traditionally used outcome measures that they have obtained from other disciplines, especially psychiatry and psychology. These include official statistics, symptom scales, and diagnostic measures. Answers to the central sociological question of how social arrangements affect mental health might require the development of explicitly sociological outcome measures. This introduction provides an overview of several issues that arise in grappling with this question. These include whether symptom scales or diagnoses best capture the mental health consequences of social arrangements; when single or multiple outcomes are necessary to compare the consequences of social arrangements across different groups; if sociologists should explore the positive as well as the negative consequences of social forces; and when sociological attention should be directed toward social-level as well as individual-level outcomes. The papers in this symposium that follow provide more detailed analyses of each of these issues. PMID- 12096697 TI - Measurement for a human science. AB - This paper argues a number of points about measurement in the sociology of mental health: (1) measurement is critical, (2) measures should represent and assess elements of human experience, taking measure of life as people feel it, sense it, and understand it, and (3) social scientists should create a human science, producing information for the people it studies so that they can better understand and control their own lives. We argue that a human science is best achieved with the use of indexes, not diagnoses, to measure mental health. We present a brief history of diagnostic instruments and detail how a diagnosis is made. We show how use of diagnoses to measure mental health discounts much human suffering. They dichotomize the true range of feelings and emotions into crude either/or distinctions that do not reflect the reality of people's lives, and they often exclude suffering such as that due to loss or illness that does not meet medical model preconceptions about mental disorder. Using diagnoses to measure mental health presents a reified image of hidden disease knowable and manageable only by trained professionals--beyond the capacity of the suffering individuals to understand and control. PMID- 12096698 TI - The categorical versus dimensional assessment controversy in the sociology of mental illness. AB - This paper reviews the sociological controversy over using categorical versus dimensional assessments in the study of psychological distress. The preference of sociologists for dimensional assessments is traced to two assumptions: (1) that the associations of predictors with psychological distress syndromes are most accurately operationalized by using dimensional measures; and (2) that no true discrete mental illnesses can reasonably be inferred to exist that would justify the creation of dichotomous measures. Methods are described in this paper to test both assumptions. An argument is made that the first assumption is the critical one and that dimensional analysis is useful only when it can be demonstrated that the associations of predictors with dimensional scores are constant throughout the relevant dimensional severity range. The paper closes with an argument for the tandem use of categorical and dimensional assessments in future sociological research. PMID- 12096699 TI - Violent behavior: a measure of emotional upset? AB - Over the past ten years, sociologists have broadened their view of what counts as an appropriate measure of mental health. This reflects a growing recognition that individuals express emotional upset in various ways. For example, some individuals are more likely to become depressed in response to stress while others are more likely to drink heavily. Contemporary studies often include measures of "internalizing" (i.e., more feeling-based measures) and "externalizing" (i.e., more behavioral measures) styles of psychopathology, especially when studying group differences in mental health. Alcohol abuse is the classic measure of externalized distress in sociological research. In this paper, we present a theoretical argument and supporting empirical evidence to argue that violent behavior should be included as a measure of externalized distress in response to stress. Our study suggests that violent behavior is a more likely response to stress among individuals with particular coping and appraisal tendencies. Specifically, violent behavior may be a more likely response to stress among individuals who tend to appraise situations as threatening while also repressing any emotional response to stress. We contend that, since some groups may be more likely than others to respond to stress with violence, it is particularly important to include measures of violent behavior when studying group difference in distress. PMID- 12096701 TI - Outcomes for the sociology of mental health: are we meeting our goals? AB - In this paper I argue that a fruitful discussion of the choice of outcomes in the sociological study of the social antecedents of mental health problems would benefit from a consideration of the goals that we are trying to achieve. The most clearly articulated goal is that of uncovering those aspects of society that produce harm. I examine the premises of the current conceptual framework--the stress paradigm--in light of its ability to fulfill this goal, and I discuss the implications for the types of outcome measures we use. PMID- 12096700 TI - The mental health continuum: from languishing to flourishing in life. AB - This paper introduces and applies an operationalization of mental health as a syndrome of symptoms of positive feelings and positive functioning in life. Dimensions and scales of subjective well-being are reviewed and conceived of as mental health symptoms. A diagnosis of the presence of mental health, described as flourishing, and the absence of mental health, characterized as languishing, is applied to data from the 1995 Midlife in the United States study of adults between the ages of 25 and 74 (n = 3,032). Findings revealed that 17.2 percent fit the criteria for flourishing, 56.6 percent were moderately mentally healthy, 12.1 percent of adults fit the criteria for languishing, and 14.1 percent fit the criteria for DSM-III-R major depressive episode (12-month), of which 9.4 percent were not languishing and 4.7 percent were also languishing. The risk of a major depressive episode was two times more likely among languishing than moderately mentally healthy adults, and nearly six times greater among languishing than flourishing adults. Multivariate analyses revealed that languishing and depression were associated with significant psychosocial impairment in terms of perceived emotional health, limitations of activities of daily living, and workdays lost or cutback. Flourishing and moderate mental health were associated with superior profiles of psychosocial functioning. The descriptive epidemiology revealed that males, older adults, more educated individuals, and married adults were more likely to be mentally healthy. Implications for the conception of mental health and the treatment and prevention of mental illness are discussed. PMID- 12096702 TI - Commentary: answers and questions in the sociology of mental health. AB - This commentary speaks to several issues that arise from the papers in this special issue. Two articles--Kessler (2002) and Mirowsky and Ross (2002)--focus on a major measurement issue: dimensional versus diagnostic-type assessments. One topic requires greater attention: the correspondence of these measures with the underlying states they supposedly measure--constructs in the psychometric tradition and empirically defined illnesses in the medical or psychiatric tradition. Conclusions about the nature of these unobserved states remain tentative at this time. Three articles--Keyes (2002), Schwartz (2002), and Umberson, Williams, and Anderson (2002)--address the expansion of mental health outcomes. The existing reliance on emotional distress is problematic for sociological research because a single disorder is not a good proxy for estimates of the overall mental health consequences of social arrangements. Although these papers present diverse and sometimes conflicting perspectives, collectively they demonstrate that no one approach to outcomes is best for all research questions. PMID- 12096703 TI - The challenge of the dependent variable. PMID- 12096704 TI - [Investigating sexual abuse of a child]. PMID- 12096705 TI - [Homeopathy after extraction of wisdom teeth]. PMID- 12096706 TI - [The danger of chronic exposure to laughing gas]. PMID- 12096707 TI - Application of a nurse-managed inpatient smoking cessation program. AB - This study reports on the effectiveness of a nurse case-managed smoking cessation program for general hospitalized patients that was continued for 3 years after clinical trials were completed. Patients admitted to the hospital who smoked were offered a smoking cessation program during their hospitalization. The program included physician advice, bedside education and counseling with a nurse specially trained in smoking cessation techniques, take-home materials (videotape, workbook, and relaxation audiotape), nicotine replacement therapy if requested or indicated, and four nurse-initiated post-discharge telephone counseling calls. Of the 2091 patients identified as smokers, 52% enrolled in the program, 18% wanted to quit on their own, 20% did not want to quit, and 10% were ineligible. The 12-month self-reported cessation rate (7-day point prevalence) was 35% if patients lost to follow-up were considered smokers, 49% if not. Patients hospitalized for cancer, cardiovascular, or pulmonary reasons were most likely to participate and had the highest self-reported cessation rates (63%, 57%, and 46%, respectively). This nurse-managed smoking cessation intervention was effective when it was put into standard hospital practice outside of its originating randomized clinical trial structure. The program, relatively inexpensive to deliver, appears to be acceptable to the majority of smokers who are hospitalized, resulted in high 1-year cessation rates, and can be extended to hospital employees and their families, work-sites, and communities on a cost recovery basis. PMID- 12096708 TI - The safety of inactivated influenza vaccine adults and children with asthma. PMID- 12096709 TI - Absence of nasal bone in fetuses with trisomy 21 at 11-14 weeks of gestation: an observational study. PMID- 12096710 TI - Randomized controlled trial of primary-school based intervention to reduce risk factors for obesity. PMID- 12096711 TI - Euro-blues. PMID- 12096712 TI - Selective dephosphorylation of the threonine(183) residue of ERK2 upon (alpha)llb(beta)3 engagement in platelets. AB - Thrombin-induced extracellular signal-regulated kinase 2 (ERK2) activation is negatively regulated in conditions of all bP3 integrin engagement and platelet aggregation. Here we show by Western blotting with antibodies against mono- and biphosphorylated forms of ERK2 that the dephosphorylation of ERK2 by alpha llb beta 3 engagement affects threonine183 and not tyrosine185. Addition of a potent serine/threonine phosphatase inhibitor, okadaic acid (OA), restored thrombin induced threonine phosphorylation of ERK2 in conditions of platelet aggregation, whereas OA had no effect in the absence of alpha llb beta 3 engagement. These observations are consistent with alpha llb beta 3 engagement acting via at least one serine/threonine phosphatase,which dephosphorylates the phosphothreonine183 residue of ERK2. Moreover, a small amount (14%) of ERK2 was translocated to the alpha llb beta 3-dependent cytoskeleton, mostly ina monophosphorylated (i.e. inactive) form, suggesting that cytoskeleton-associated ERK2 plays only a minor role, if any. Finally, we show that negative regulation (i.e. dephosphorylation)occurs primarily or totally in the cytosol and that the alpha llb beta 3-dependent ERK2 Thr183-specific phosphatase is different from phosphatase 1 (PP1) or PP2A. We conclude that all alpha llb beta 3 engagement down-regulates ERK2 through selective dephosphorylation of the phosphothreonine183 residue by a cytosolic serine/threonine phosphatase different from known platelet phosphatases. PMID- 12096713 TI - Activation of Pyk2/RAFTK induces tyrosine phosphorylation of alpha-synuclein via Src-family kinases. AB - alpha-Synuclein (alpha S) is a neuronal protein that has been implicated in the pathogenesis of Parkinson's disease. The present report demonstrates that the protein tyrosine kinase Pyk2/RAFTK is involved in cell stress-induced tyrosine phosphorylation of alpha S. Hyperosmotic stress induced tyrosine phosphorylation of alpha S via Pyk2/RAFTK at tyrosine residue 125. Pyk2/RAFTK-mediated phosphorylation of alpha S was primarily achieved with Src-family kinases. In addition, osmotic stress-induced phosphorylation of alpha S was dependent on Pyk2/RAFTK activation. Accordingly, such results indicate that Pyk2/RAFTK lies upstream of Src-family kinases in the signaling cascade by which osmotic stress induces tyrosine phosphorylation of alpha S. PMID- 12096714 TI - RNA interference (RNA(i)) induction with various types of synthetic oligonucleotide duplexes in cultured human cells. AB - Various types of synthetic oligonucleotide duplexes against the Photinus luciferase gene were tested on their induction of the sequence-specific RNA interference (RNAi) activity in transfected human cells. Results indicate that RNA duplexes with ribonucleotide 3' overhangs rather than those with deoxyribonucleotide 3' overhangs induce more efficient RNAi activity, and that sense-stranded DNA/antisense-stranded RNA hybrids induce a moderate RNAi activity. These results suggest that there is a difference in the potential of oligonucleotide duplexes to be RNAi mediators, i.e. short interfering RNAs (siRNAs), between human RNAi and invertebrate RNAi. The data further show that different siRNAs induce different levels of RNAi. PMID- 12096715 TI - A patient on hydroxyurea for sickle cell disease who developed an opportunistic infection. PMID- 12096716 TI - FLT3 internal tandem duplication mutations are rare in agnogenic myeloid metaplasia. PMID- 12096717 TI - Platelet-dependent action of high-dose factor VIIa. PMID- 12096718 TI - Cyclosporine modulation in poor-risk acute myeloid leukemia. PMID- 12096720 TI - Current readings in nuclear medicine. PMID- 12096719 TI - Binding of imatinib by alpha(1)-acid glycoprotein. PMID- 12096722 TI - Should new drugs be avoided? PMID- 12096721 TI - Can fruit juices cause irritable bowel syndrome?...and could they play a role in colic? PMID- 12096723 TI - . . . And just how safe is Echinacea? PMID- 12096724 TI - New vaccine will help ease shortages. PMID- 12096725 TI - How to dispose of mercury thermometers. PMID- 12096726 TI - Lyme disease season is here...there's no vaccine...but there are ways to prevent it. PMID- 12096727 TI - Product recalls. Harry Potter key chains. PMID- 12096728 TI - Product recalls. Infant car seats/carriers. PMID- 12096729 TI - 3D imaging in private practice. PMID- 12096730 TI - Influence of vision on head stabilization strategies in older adults during walking. AB - BACKGROUND: Maintaining balance during dynamic activities is essential for preventing falls in older adults. Head stabilization contributes to dynamic balance, especially during the functional task of walking. Head stability and the role of vision in this process have not been studied during walking in older adults. METHODS: Seventeen older adults (76.2 +/- 6.9 years) and 20 young adults (26.0 +/- 3.4 years) walked with their eyes open (EO), with their eyes closed (EC), and with fixed gaze (FG). Participants performed three trials of each condition. Sagittal plane head and trunk angular velocities in space were obtained using an infrared camera system with passive reflective markers. Frequency analyses of head-on-trunk with respect to trunk gains and phases were examined for head-trunk movement strategies used for head stability. Average walking velocity, cadence, and peak head velocity were calculated for each condition. RESULTS: Differences between age groups demonstrated that older adults decreased walking velocity in EO (p =.022). FG (p = .021), and EC (p = .022). and decreased cadence during EC (p = .007). Peak head velocity also decreased across conditions (p < .0001) for older adults. Movement patterns demonstrated increased head stability during EO. diminished head stability with EC, and improved head stability with FG as older adult patterns resembled those of young adults. CONCLUSIONS: Increased stability of the lower extremity outcome measures for older adults was indicated by reductions in walking velocity and cadence. Concomitant increases in head stability were related to visual tasks. Increased stability may serve as a protective mechanism to prevent falls. Further, vision facilitates the head stabilization process for older adults to compensate for age related decrements in other sensory systems subserving dynamic balance. PMID- 12096731 TI - Congressional testimony 2002: nurse scientists face the challenge. PMID- 12096733 TI - Need to clarify start of patient advocacy. PMID- 12096734 TI - Nurses must take time to talk to their patients. PMID- 12096735 TI - Closure of care homes must stop now. PMID- 12096737 TI - The essential oil secretory structures of Prostanthera ovalifolia (Lamiaceae). AB - The structure of the essential oil secretory tissues of Prostanthera ovalifolia R.Br was investigated using bright- and dark-field optical microscopy, and scanning and transmission electron microscopy. The leaves of P. ovalifolia have glandular trichomes of the peltate type common to many Lamiaceae species. The trichomes consist of a basal cell embedded in the epidermis, a stalk cell with heavily cutinized walls and a 16-celled secretory head, but they differ from those of many previously reported Lamiaceae species in their morphological form defined by the elevated cuticle. The sub-cuticular space contains a mixture of lipid and aqueous phases. Secretory cells have dense cytoplasm with many leucoplasts present. Volatile terpenoids are eliminated from the cytoplasm into the sub-cuticular space, the site of essential oil accumulation, via granulocrine secretion. PMID- 12096736 TI - Altering plant-microbe interaction through artificially manipulating bacterial quorum sensing. AB - Many bacteria regulate diverse physiological processes in concert with their population size. Bacterial cell-to-cell communication utilizes small diffusible signal molecules, which the bacteria both produce and perceive. The bacteria couple gene expression to cell density by eliciting a response only when the signalling molecules reach a critical threshold (a point at which the population is said to be 'quorate'). The population as a whole is thus able to modify its behaviour as a single unit. Amongst Gram-negative bacteria, the quorum sensing signals most commonly used are N-acylhomoserine lactones (AHLs). It is now apparent that AHLs are used for regulating diverse behaviours in epiphytic, rhizosphere-inhabiting and plant pathogenic bacteria and that plants may produce their own metabolites that interfere with this signalling. Transgenic plants that produce high levels of AHLs or which can degrade bacterial-produced AHLs have been made. These plants have dramatically altered susceptibilities to infection by pathogenic Erwinia species. In addition, such plants will prove useful tools in determining the roles of AHL-regulated density-dependent behaviour in growth promoting, biological control and pathogenic plant-associated bacterial species. PMID- 12096738 TI - Performance of an age series of alnus-cardamom plantations in the Sikkim Himalaya: productivity, energetics and efficiencies. AB - Biomass, net primary productivity, energetics and energy efficiencies were estimated in an age series of Alnus-cardamom plantations in the eastern Himalaya. The impact of stand age (5, 10, 15, 20, 30 and 40 years) on the performance of mixtures of N2-fixing (Alnus nepalensis) and non-N2-fixing (large cardamom) plants was studied. Large cardamom (Amomum subulatum) is the most important perennial cash crop in the region and is cultivated predominantly under Alnus trees. Net primary productivity was lowest (7 t ha(-1) per year) in the 40-year old stand and was more than three times higher (22 t ha(-1) per year) in the 15 year-old stand. Agronomic yield of large cardamom peaked between 15 and 20 years of age. Cardamom productivity doubled from the 5- to the 15-year-old stand, and then decreased with plantation age to reach a minimum in the 40-year-old stand. Performance of cardamom in association of N2-fixing Alnus remained beneficial until 20 years of age. Annual net energy fixation was highest (444 x 10(6) kJ ha( 1) per year) in the 15-year-old stand, being 1.4 times that of the 5-year-old stand and 2.9-times that of the 40-year-old stand. Inverse relationships of production efficiency, energy conversion efficiency and energy utilized in N2 fixation against stand age, and a positive relationship between production efficiency and energy conversion efficiency suggest that the younger plantations are more productive. The Alnus-cardamom plantation system will be sustainable by adopting a rotational cycle of 15 to 20 years. PMID- 12096739 TI - Performance of an age series of Alnus-cardamom plantations in the Sikkim Himalaya: nutrient dynamics. AB - Nutrient cycling, nutrient use efficiency and nitrogen fixation in an age series of Alnus-cardamom plantations were studied in the eastern Himalaya. The impact of stand age (5, 10, 15, 20, 30 and 40 years) on the nutrient dynamics of mixtures of N2-fixing (Alnus nepalensis) and non-N2-fixing (large cardamom) plants was assessed. Foliar nutrient concentrations of Alnus decreased with advancing age groups of plantations and showed an inverse relationship with stand age. Annual N fixation increased from the 5-year-old stand (52 kg ha(-1)), peaking in the 15 year-old stand (155 kg ha(-1)) and then decreased with increasing plantation age. Nitrogen and phosphorus uptake was lowest in the 40-year-old stand, and highest in the 15- and 5-year-old stand, respectively. Nutrient storage in understorey cardamom was very high: up to 31 % N and 59 % P of the stand total in the 15-year old stand. Nutrient use efficiency was higher (with faster turnover times) in younger stands and decreased (with slower turnover times) in older plantations. Nitrogen retranslocation showed a strong positive relationship with stand age, while that of P was inversely related to stand age. Nutrient standing stock, uptake and return were also highest in the 15-year-old stand. Nitrogen and P cycling in Alnus-cardamom plantations was functionally balanced. Nutrient cycling and dynamics indicated that Alnus-cardamom plantations performed sustainably up to 15-20 years. The management practice should be altered to incorporate replantation after this age. PMID- 12096740 TI - Variation amongst survivor populations of white clover collected from sites across Europe: growth attributes and physiological responses to low temperature. AB - Experiments were carried out at IGER, Aberystwyth, UK to investigate traits of direct relevance to the processes of overwintering and spring growth in white clover (Trifolium repens L.). The plant material used was derived from baseline populations of the cultivar AberHerald and survivor populations generated after 2 3 years' growth in Germany (Kiel), Sweden (Uppsala) and Switzerland (Zurich). The aims of the experiments were to measure the level of genetic shift that had occurred in certain traits due to selection in the survivor populations by comparing these with the baseline population. The adaptive significance of traits was assessed by determining the extent to which stabilizing selection had operated to reduce levels of intra-population variation. Significant differences were found in the responses of leaf production to two temperature treatments in the survivor populations from Germany and Sweden compared with the Swiss and baseline material. Plants of the former two populations produced much more leaf than the others at the higher temperature, but leaf production rates at the lower temperature did not differ. As this experiment used cloned genotypes in the two treatments, the result suggests that a higher degree of phenotypic plasticity for this trait had been selected for in the German and Swedish populations. These populations also showed greater rates of regrowth of leaves from terminal buds exposed to sub-zero temperatures, but there were no differences between populations in levels of freezing tolerance, or in stolon carbohydrate content. Genetic shift occurred in the degree of unsaturation of stolon lipids, with all three survivor populations possessing higher proportions of unsaturated fatty acids than the baseline. Stabilizing selection also operated on this trait in the survivor populations, suggesting that it is of adaptive significance in cool climates. PMID- 12096741 TI - Detection of root mucilage using an anti-fucose antibody. AB - Plant root mucilage is known to enhance soil quality by contributing towards the soil carbon pool, soil aggregation, detoxification of heavy metal ions and interactions with rhizospheric microflora. Mucilage consists of many monosaccharide units, including fucose which can be used as an indicator for plant root based polysaccharides. This is the first report of an immunological technique developed to use anti-fucose antibodies as markers for probing and localizing fucosyl residues in mucilage polysaccharide and, in turn, for localization of plant root mucilage. Fucose was complexed with bovine serum albumin to raise antibodies against fucose. A fucose-directed antibody was shown to cross-react with root cap mucilages from grasses. This antibody was used to localize root mucilage polysaccharide in maize and wheat root caps using immunogold electron microscopy. Abundant labelling could be localized on the cell wall, and in the intercellular matrix and vesicles of the peripheral root cap cells. Labelling was less intense in cells towards the centre of the root cap tissue. Control experiments confirmed that immunogold localization of fucose was specific and reliable. PMID- 12096742 TI - Spatial arrangement of branches in relation to slope and neighbourhood competition. AB - To gain a better understanding of the effects of spatial structure on patterns of neighbourhood competition among hardwood trees, the three-dimensional extension of primary branches was surveyed for ten community-grown Castanea crenata (Fagaceae) trees with respect to the positioning of neighbouring branches and the slope of the forest floor. There were significantly more branches extending towards the lower side of the slope than towards the upper side, but structural properties such as branch length and vertical angle were not affected by slope. When horizontal extension of a branch towards its neighbour was compared for a C. crenata branch and a neighbouring heterospecific, the former was significantly narrower than the latter when the inter-branch distance (horizontal distance between the base positions of two neighbouring branches) was short (< approx. 5 m). Castanea crenata branches tended to extend in a direction avoiding neighbouring branches of heterospecifics when the inter-branch distance was short. Furthermore, for an inter-branch distance <3 m, the horizontal extension of a C. crenata branch was less when it was neighbouring a heterospecific branch than when neighbouring a conspecific branch. These results suggest that horizontal extension of C. crenata branches is more prone to spatial invasion by nearby neighbouring branches of heterospecifics, and that the invasion can be lessened when C. crenata trees are spatially aggregated. The reason why such an arrangement occurs is discussed in relation to the later leaf-flush of C. crenata compared with that of other species in the forest. PMID- 12096743 TI - Variability in thermal response of primed and non-primed seeds of squirreltail [Elymus elymoides (Raf.) Swezey and Elymus multisetus (J. G. Smith) M. E. Jones]. AB - Bottlebrush squirreltail [Elymus elymoides (Raf.) Swezey = Sitanion hystrix (Nutt.) J. G. Smith] and big squirrel-tail [Elymus multisetus (J. G. Smith) M. E. Jones = Sitanion jubatum (J. G. Smith)] have a broad geographical distribution and have been identified as high priority species for restoration of degraded rangelands in the western United States. These rangelands exhibit high annual and seasonal variability in seedbed microclimate. The objective of this study was to examine variability in thermal response of both primed and non-primed seeds of these species in the context of field-variable temperature regimes. Seed priming treatments were selected to optimize germination rate in a low-temperature test environment. Primed and non-primed seeds were evaluated for laboratory germination response under 12 constant temperature treatments between 3 and 36 degrees C. Thermal time and base temperature were estimated by regression analysis of germination rate as a function of temperature in the sub-optimal temperature range. The thermal germination model and 6 years of field temperature data were used to simulate the potential germination response under different field planting scenarios. Seed priming reduced the total germination percentage of some seedlots, especially at higher germination temperatures. Seed priming increased the germination rate (reduced the number of days to 50 % germination) by 3.8-8.4 d at 6 degrees C with a mean germination advancement of 6.9 +/- 0.6 d. Maximum germination advancement in the model simulations was 5-10 d for planting dates between I March and 15 May. Model simulations can be used to expand germination analysis beyond simple treatment comparisons, to include a probabilistic description of potential germination response under historical or potential future conditions of seedbed microclimate. PMID- 12096744 TI - Testing taxonomic and biogeographical relationships in a narrow Mediterranean endemic complex (Hippocrepis balearica) using RAPD markers. AB - Analyses of RAPD profiles from 17 populations of the Hippocrepis balearica complex revealed a highly structured geographic pattern, not only among continental-insular areas but also within the eastern Balearic islands. In marked contrast to previous morphometric results, a clear separation between continental and insular samples was found, and intermediates between H. balearica and H. valentina samples were not detected. Molecular data indicated that western and eastern Balearic populations of the complex (H. grosii and H. balearica) were more closely related to each other than to continental populations (H. valentina). Multivariate analyses of the RAPD data clearly indicated that the similarities between continental and eastern Balearic samples of the H. balearica complex recovered by morphometric methods are due either to parallel evolution or to retention of plesiomorphic features. PMID- 12096745 TI - Physiological ecology of Mesozoic polar forests in a high CO2 environment. AB - Fossils show that coniferous forests extended into polar regions during the Mesozoic, a time when models and independent paleo-CO2 indicators suggest that the atmospheric CO2 concentration was at least double that of the present day. Consequently, such polar forests would have experienced high CO2 interacting with an extreme variation in light. Here we describe an experiment investigating this plant-environment interaction for extant tree species that were important components of polar forests, and give results from the first year of treatment. Specifically, we tested the hypotheses that growth in elevated CO2 (1) stimulates photosynthesis; (2) reduces photoinhibition during the polar summer; and (3) reduces respiration of above- and below-ground plant organs. Our results indicate that CO2 fertilization generally does not affect photosynthesis under continuous daylight characteristic of the polar summer but does increase it when the period of illumination is shorter. Growth in elevated CO2 did not alter the potential for photoinhibition. CO2 enrichment significantly reduced leaf and root respiration rates by 50 and 25 %, respectively, in a range of evergreen taxa. Incorporating these observed CO2 effects into numerical simulations using a process-based model of coniferous forest growth indicates that a high paleo-CO2 concentration would have increased the productivity of Cretaceous conifer forests in northern Alaska. This results from decreased respiratory costs that more than compensate for the absence of high CO2-high temperature interactions during the polar summer. The longer-term effects of CO2 enrichment on seasonal changes in the above- and below-ground carbon balance of trees are discussed. PMID- 12096747 TI - Inside "The Lunatic Manufacturing Company': the persecuted world of John Gilmour. PMID- 12096746 TI - The development of potential screens based on shoot calcium and iron concentrations for the evaluation of tolerance in Egyptian genotypes of white lupin (Lupinus albus L.) to limed soils. AB - European cultivars of white lupin (Lupinus albus L.) grow poorly in limed or calcareous soils. However, Egyptian genotypes are grown successfully in highly calcareous soil and show no stress symptoms. To examine their physiological responses to alkaline soil and develop potential screens for tolerance, three experiments were conducted in limed and non-limed (neutral pH) soil. Measurements included net CO2 uptake, and the partitioning of Fe2+ and Fe3+ and soluble and insoluble Ca in stem and leaf tissue. Intolerant plants showed clear symptoms of stress, whereas stress in the Egyptian genotypes and in L pilosus Murr. (a tolerant species) was less marked. Only the intolerant plants became chlorotic and this contributed to their reduced net CO2 uptake in the limed soil. In contrast, Egyptian genotypes and L pilosus showed no change in net CO2 uptake between the soils. The partitioning of Ca and Fe either resulted from the stress responses, or was itself a stress response. L pilosus and some Egyptian genotypes differed in soluble Ca concentrations compared with the intolerant cultivars, although no significant difference was apparent in the Ca partitioning of the Egyptian genotype Giza 1. In a limed soil, Giza 1 maintained its stem Fe3+ concentration at a level comparable with that of plants grown in non-limed soil, whereas stem [Fe3+] of an intolerant genotype increased. Gizal increased the percentage of plant Fe that was Fe2+ in its leaf tissue under these conditions; that of the intolerant genotype was reduced. The potential tolerance of the Egyptian genotypes through these mechanisms and the possibility of nutritional based screens are discussed. PMID- 12096748 TI - Constructing curative instruments: psychiatric architecture in Norway, 1820-1920. PMID- 12096749 TI - An English psychiatrist in Australia: memories of Eric Cunningham Dax and the Victorian Mental Hygiene Authority, 1951-1969. PMID- 12096750 TI - Emil Kraepelin's 'Self-Assessment': clinical autography in historical context. AB - Contemporary psychiatrists and historians know very little about the life of Emil Kraepelin. Until recently they had to glean what information they could from his memoirs, which had more to say about his travels to far-flung corners of the world than about his own mental life. Now, however, a unique historical document has been uncovered which opens a window on Kraepelin's own psyche. In a manuscript written shortly after World War I, Kraepelin turned his diagnostic methods on himself and clinically documented his own state of mind. This paper reproduces a translation of Kraepelin's 'Self-Assessment' and places it in its historical context. PMID- 12096751 TI - 1H NMR study of the binding of Bis(acridines) to d(AT)5.d(AT)5. 1. Mode of binding. AB - 1H NMR has been used to investigate the mode of binding to d(AT)5.d(AT)5 of a series of bis(acridine) derivatives connected by different types of linker chains. The length and character (ionic, aliphatic, rigid, and flexible) of the linker chains are found to have a profound effect on the binding of these derivatives to the DNA. Bis(acridine) derivatives with linker chains shorter than 9 A monointercalate under the conditions used in the NMR study, whereas those bis(acridines) with chains of 9.8 A or longer bisintercalate. We find no evidence for the violation of the so-called neighbor exclusion principle. Although all of the bis(acridines) contain the same chromophores, their NMR spectra clearly demonstrate that they form complexes with d(AT)5.d(AT)5 which have different structures. This emphasizes the important effect that the linker chain has on the structure of the intercalation complex. PMID- 12096752 TI - Applications of therapeutic jurisprudence in rehabilitation for people with severe and disabling mental illness. PMID- 12096753 TI - Civil commitment laws: an analysis and critique. PMID- 12096754 TI - 1H NMR study of the binding of bis(acridines) to d(AT)5.d(AT)5. 2. Dynamic aspects. AB - Measurements of the 1H NMR spectra and relaxation rates were used to study the dynamic properties of 9-aminoacridine (9AA) and four bis(acridine) complexes with d(AT)5.d(AT)5. The behavior of the 9AA (monointercalator) and that of C8 (bisintercalator containing an eight-carbon atom linker chain) are entirely similar. For both compounds, the lifetime of the drug in a particular binding site is 2-3 ms at approximately 20 degrees C, and neither affects the A.T base pair opening rates. The complex with C10 (bisintercalator containing a 10-carbon atom linker chain) is slightly more stable than the C8 complex since its estimated binding site lifetime is 5-10 ms at 29 degrees C. Base pairs adjacent to the bound C10 are destabilized, relative to free d(AT)5.d(AT)5, but other base pairs in the C10 complex are little affected. Bis(acridine) pyrazole (BAPY) and bis(acridine) spermine (BAS) considerably stabilize those base pairs that are sandwiched between the two acridine chromophores, but in the BAS complex proton exchange from the two flanking base pairs appears to be accelerated, relative to free d(AT)5.d(AT)5. The lifetime of these drugs in specific binding sites is too long (>10 ms) to be manifested in increased line widths, at least up to 41 degrees C. An important conclusion from this study is that certain bisintercalators rapidly migrate along DNA, despite having large binding constants (K>10(6) M-1). For C8 and C10 complexes, migration rates are little different from those deduced for 9AA. The rigid linker chain in BAPY and the charge interactions in BAS retard migration of these two bisintercalators. These results provide new parameters that are useful in understanding the biochemical and biological properties of these and other bisintercalating drugs. PMID- 12096755 TI - Continuous glucose monitoring in the management of diabetes mellitus. AB - Continuous glucose monitors (CGMs) measure interstitial fluid (ISF) glucose, and provide information about continuous glucose fluctuations that is not otherwise captured by intermittent blood glucose testing. CGMs may benefit patients having difficulty controlling their blood sugar or during initiation or monitoring of insulin pump use. CGMs require calibration with finger-stick tests and supplement, but do not replace conventional blood glucose testing. CGM values correspond to blood glucose values taken approximately 13-18 minutes earlier and may differ from metered readings. PMID- 12096756 TI - Perceived orientation in physical and virtual environments: changes in perceived orientation as a function of idiothetic information available. AB - Two experiments examined perceived spatial orientation in a small environment as a function of experiencing that environment under three conditions: real-world, desktop-display (DD), and head-mounted display (HMD). Across the three conditions, participants acquired two targets located on a perimeter surrounding them, and attempted to remember the relative locations of the targets. Subsequently, participants were tested on how accurately and consistently they could point in the remembered direction of a previously seen target. Results showed that participants were significantly more consistent in the real-world and HMD conditions than in the DD condition. Further, it is shown that the advantages observed in the HMD and real-world conditions were not simply due to nonspatial response strategies. These results suggest that the additional idiothetic information afforded in the real-world and HMD conditions is useful for orientation purposes in our presented task domain. Our results are relevant to interface design issues concerning tasks that require spatial search, navigation, and visualization. PMID- 12096757 TI - The myth of Freud's ostracism by the medical community in 1896-1905: Jeffrey Masson's assault on the truth. AB - The story recounted by Jeffrey M. Masson of the medical community's outraged response to the seduction theory is treated as historical fact in some of the recent literature on Freud's early psychoanalytic experiences. In this article the evidence adduced by Masson in The Assault on Truth (1984) to buttress his account of Freud's supposed ostracism is critically examined. It is concluded that this evidence fails to substantiate Masson's version of events, that there is abundant evidence that refutes it, and that he has ignored the historical research that demonstrates that the notion that Freud's early psychoanalytic writings received an irrationally hostile reception is a myth. PMID- 12096758 TI - Between the laboratory and life: child development research in Toronto, 1919 1965. AB - Between 1919 and 1956, psychologists at the University of Toronto built a research program in developmental psychology on a functionalist, holistic, and ecological basis. They conducted longitudinal studies on mental health in growing children in educational settings instead of in laboratories and formed strong alliances with the local educational system in order to do so. They initially defined mental health as adjustment and considered conditions within schools to be conducive to its attainment. After developing a psychological theory of personality development, they came to view educational conditions as discouraging the development of mental health. The alliance between the educational system and psychology consequently unraveled, and the program declined. PMID- 12096759 TI - The Hawthorne experiments and the introduction of Jean Piaget in American industrial psychology, 1929-1932. AB - The Hawthorne interview program between 1929 and 1932 was one of the most significant industrial studies in the United States. The Hawthorne researchers applied Jean Piaget's clinical method in their extensive interviews with tens of thousands of workers. Chiefly responsible for the program's methodology was Elton Mayo, an Australian who saw interviewing as a means to promote social cooperation. Previous discussions of the Hawthorne experiments have ignored the influence of Piaget in the social sciences. This article provides an account of Mayo's and the Hawthorne researchers' efforts to fuse Piaget's innovation with burgeoning American industrial psychology. The endeavor was not an isolated event but rather drew on the theories and practice of Janet-Piaget psychology, on the support of the Laura Spelman Rockefeller Memorial Foundation, and on the discourse among social scientists about Piaget's work. PMID- 12096760 TI - Psychologists and the national socialist access to power. AB - The destruction of psychological science during the early years of the National Socialist (NS) regime is an important part of psychology's history. The proceedings of the congresses of the German Society for Psychology describe the very quick takeover of German psychology by the supporters of the regime and the removal of any opponents of the regime and of Jewish members from the executive structure. The leadership of the field was taken over by psychologists who were early supporters of the government's antidemocratic and anti-Semitic policies. Some of the figures of the NS period reappeared in leading positions in the postwar period. PMID- 12096761 TI - Helmut E. Adler (1920-2001). PMID- 12096762 TI - Paul T. Mountjoy (1924-2001). PMID- 12096763 TI - Edward J. Haupt (1936-2001). PMID- 12096764 TI - 1115 ways to waive Medicaid and SCHIP rules. AB - This issue brief explores the history and context of the Section 1115 Medicaid waiver authority, discusses the Health Insurance Flexibility and Accountability (HIFA) initiative under way in the Bush administration, and considers some of the potential impacts that HIFA could have on state budgets and access to health care for low-income families. Finally, it considers the future of Section 1115 waivers as a vehicle for modifying Medicaid and the State Children's Health Insurance Program. PMID- 12096765 TI - Welfare reform and American Indians: critical issues for reauthorization. AB - With reauthorization of the Personal Responsibility and Work Opportunity Reconciliation Act (P.L. 104-193) under consideration in Congress, this issue brief examines some key features of the welfare reform legislation as it applies to American Indians and reviews its impact on this population. It looks at the flexibility the act has given American Indian tribes to design and operate their own Temporary Assistance for Needy Families programs and explores barriers they face in doing so. The paper also examines some of the policy issues and opportunities surrounding welfare reform for American Indians. PMID- 12096766 TI - Managing advanced illness: a quality and cost challenge to Medicare, Medicaid, and private insurers. AB - This background paper examines approaches to delivering and financing health services for persons with advanced chronic illness. Recognizing that the Medicare program is the major insurer of such services and that its share is likely to increase due to the aging of the population, the paper focuses on the nature and structure of Medicare hospice-and, to a very limited extent, palliative-care benefits. It also explores Medicaid benefits, as well as coverage through private insurance, including indemnity, point-of-service,and preferred-provider organization products. In addition to giving descriptive information on the various programs, the paper looks at quality concerns and cost and payment considerations. PMID- 12096767 TI - Writing the Victorian City: discourses of risk, connection, and inevitability. PMID- 12096768 TI - Trauma, memory, and railway disaster: the Dickensian Connection. PMID- 12096769 TI - Continued challenges for the physician executive in managed care. PMID- 12096770 TI - The future of American healthcare: a mandatory change to mass customization. PMID- 12096771 TI - Keep a story in your heart: a message to the class of 2002. PMID- 12096772 TI - Engendering community: learning and sharing expertise in the skills and practices of teaching. PMID- 12096773 TI - The process of seeking stress-care: coping as experienced by senior baccalaureate nursing students in response to appraised clinical stress. AB - High levels of clinical stress experienced by nursing students continue to be identified in the literature. However, much is still unexplored about the coping processes of nursing students in response to appraised clinical stress. A grounded theory methodological approach was used to acquire a substantive theory of the processes through which senior baccalaureate nursing students manage the demands of the clinical person-environment relationships that are appraised as stressful and the emotions they generate. A theoretical sample of 16 volunteer senior generic nursing students from two baccalaureate nursing programs was determined by saturation of the data. Each participant was interviewed twice. The basic social problem experienced by these senior baccalaureate nursing students was the chaos created in their lives while trying to manage the demands of the appraised clinical stress (i.e., the clinical person-environment relationship). The three-stage process of seeking stress- care, including encountering changing self, loss of self, and regaining managed self, helped the students manage this chaos. Each of the stages related to the processes that transpired within the students as they dealt with the chaos created in their life while trying to manage the demands of appraised clinical stress. Implications include the development of the stress-care construct and innovative instructor interventions in the clinical setting. PMID- 12096774 TI - Health behaviors of nursing students: a longitudinal study. AB - To ascertain the degree to which a health science curriculum may influence health related behaviors among students, nine selected health behaviors of 52 undergraduate nursing students were studied for 3 consecutive years and compared to those of a similar group of education students. In addition, the health behaviors of both student groups were compared to those of the general population. During the study, the nursing students had no significant alterations in the observed health behaviors, and no significant differences were observed between the health behaviors of the two groups of students. Both groups manifested certain health behaviors that were significantly different from the general population. However, nursing students were observed more frequently to be significantly different from the general population. A 3-year nursing curriculum may have facilitated the adoption of some personal health behaviors, but the time demands of academic life may have induced students to reduce time-consuming health behaviors, such as getting enough sleep, eating breakfast, and exercising. PMID- 12096776 TI - The NCLEX-RN experience: qualitative interviews with graduates of a baccalaureate nursing program. AB - It is important for nursing faculty to pay attention to individual as well as institutional results on the National Council Licensure Examination for RNs (NCLEX-RN). This study was designed to identify themes to help faculty understand the NCLEX-RN experience from students' perspectives and help future students pass the examination at the first sitting. A sample of 1998 graduates of a baccalaureate program was selected, which included 10 students who were successful and 9 who were unsuccessful on the first testing attempt. Participants were interviewed about the testing experience and the relationship between nursing education and the NCLEX-RN. Findings indicate that participants who passed on the first attempt accepted responsibility for learning, were proactive in test preparation, took the examination when they felt ready, and used stress management techniques to cope with this challenge. The unsuccessful participants tended to perceive their lack of success on the NCLEX-RN was the responsibility of others, seemed less able to manage stress, and took the examination when they did not feel ready. Both successful and unsuccessful participants felt unprepared to answer NCLEX-RN-type questions and believed nothing had prepared them for this experience. PMID- 12096775 TI - Perceptions of the maternal-child clinical rotation: the male student nurse experience. AB - Although male student nurses have expressed their discomfort with beginning their maternal-child clinical experience, little actually is known about the nature and quality of their experience. To address this gap, a phenomenological study was conducted to investigate the experiences of 8 male student nurses who had completed their maternal-child rotation. Three theme clusters emerged: preconceptions about the maternal-child rotation, enduring the clinical experience, and surviving the clinical rotation. Students began their rotations with mixed feelings (e.g., dread, anxiety, apprehension, unknowing, fear of rejection). Maternal-child nursing was viewed by male students as a woman's domain, in which they were visitors. The students expressed concerns about meeting clinical objectives and personal goals because of their gender and possible misinterpretations of their care. Consequently, they undertook special precautions when performing postpartum assessments. Faculty played a significant role in clients' responses to the students and in providing a supportive learning environment. Faculty need to be cognizant of students' feelings as they plan clinical experiences. Identification of student concerns prior to the experience may be beneficial for enhancing the learning environment. PMID- 12096777 TI - Maintaining professional boundaries: preparing nursing students for the challenge. PMID- 12096778 TI - Professional development: assuring growth of RN-to-BSN students. PMID- 12096779 TI - Call for revolution: a new approach to describing allograft deterioration. AB - I propose a set of definable entities in the renal transplant course, eliminating the need for the term 'chronic rejection'. The status of a renal transplant can be defined by the presence and extent of rejection (T-cell-mediated or antibody mediated); allograft nephropathy (parenchymal atrophy, fibrosis, and fibrous intimal thickening in arteries); transplant glomerulopathy; specific diseases; and factors which could accelerate progression. The level of function and the slope of the loss of function should be separately determined. This approach can be applied both in research and in clinical practice, and can be adapted to other organ transplants. PMID- 12096780 TI - Aortic interposition grafts: a model of rejection or rejection of the model? PMID- 12096781 TI - Fibrosis and atrophy in the renal allograft: interim report and new directions. AB - The topic of chronic allograft nephropathy/chronic rejection is reviewed, with focus on fibrosing/sclerosing changes and late loss of function in renal allografts. Discussion includes a review of the problem, pathological and clinical findings, and new directions. Emphasis is placed on definition of specific diagnostic entities in these allografts, and identification of ongoing/active processes in this setting that might be amenable to direct intervention. A schema for categorizing these cases is proposed. PMID- 12096782 TI - Recipient cells form the intimal proliferative lesion in the rat aortic model of allograft arteriosclerosis. AB - Chronic rejection is the leading cause of late graft loss following solid organ transplantation and is characterized by a vasculopathy referred to as allograft arteriosclerosis. While the etiology of allograft arteriosclerosis remains unknown, it has been hypothesized that migration of donor medial smooth muscle cells into the intimal compartment is responsible for the formation of the occlusive lesion (neointima). In this study we have used aortic interposition grafts between fully histoincompatible rat strains (Brown Norway and Lewis) to investigate the origin of the neointimal cells. Three transplant paradigms were used: BN to Lew, Lew to BN and BN to Lew with immunosuppression. Neointimal cells were isolated from aortic transplant tissue through an EDTA wash/mechanical stripping technique. We have developed polymerase chain reaction primers to the MHC1 allele that are specific to each rat strains' DNA. Polymerase chain reaction analysis, using the strain-specific primers and purified neointimal cell DNA from transplanted aortic tissue from all three experimental groups, demonstrated that the neointimal cells are of recipient, and not donor origin. PMID- 12096783 TI - Activation-induced expression of cell surface CD28 on mouse T lymphocytes is inhibited by cyclosporine A. AB - T-cell activation requires both T-cell receptor signaling and a costimulatory signal provided by CD28 which enhances and prolongs interleukin-2 (IL-2) production. To determine the effect of cyclosporine A (CsA) on constitutive and activation-induced CD28 expression, mouse T cells were exposed to CsA (0.1 microM) in the absence or presence of anti-CD3 monoclonal antibody (mAb). CD28 expression was then determined by flow cytometry. CsA treatment prevented activation-induced CD28 expression but did not affect constitutive CD28 expression. Inhibition of inducible CD28 expression by CsA was not rapidly reversible, requiring 48h of restimulation in the absence of CsA for CD28 expression to return to control levels. T cells activated in the presence of combined anti-IL-2 and anti-CD25 mAb (both 10 microg/mL) also exhibited reduced CD28 expression, suggesting that activation-induced CD28 expression is, at least in part, an IL-2-dependent process. However, the inhibitory effect of CsA on activation-induced CD28 expression was maintained in the presence of exogenous IL 2 (250 U/mL). We conclude that CsA, by inhibiting activation-induced expression of costimulatory CD28 molecules by T lymphocytes, may interfere with the ability of CD28 to provide an optimal costimulatory signal for sustained IL-2 production following T-cell activation. PMID- 12096784 TI - Prevention of acute lung allograft rejection in rat by CTLA4Ig. AB - CTLA4 immunoglobulin (CTLA4Ig), which binds with a high affinity to B7-1 and B7 2, interrupts T-cell activation by inhibiting costimulatory signal. CTLA4Ig has been used in hopes of achieving antigen-specific tolerance induction in several solid organ transplants. In lung allograft rejection, however, its use has been controversial in terms of its effect on prevention of rejection. In the present study, the effect of murine CTLA4Ig on rat-lung allograft rejection was investigated. Rat left-lung transplantation was performed in an RT1 incompatible donor (Brown Norway; BN)-recipient (F344) combination. All allografts (n = 12) without any treatment were rejected within 7 days after transplantation. A single injection of murine form CTLA41g at a dose of 100 microg intraperitoneally (ip) or intravenously (iv) on day 1 post-transplantation achieved long-term graft survival (>90days) in 2/5 (40%) and 3/8 (38%), respectively. Moreover, 6/7 (86%) allografts in rats that received iv injection of 500 microg CTLA4Ig survived more than 90days. Allograft survival in the CTLA4Ig 500 microg iv recipient group was significantly longer than that in the no-treatment control or control immunoglobulin group (p <0.01). Four out of seven recipients bearing functional allografts for more than 90 days with the CTLA4Ig treatment accepted donor specific skin grafts, whereas all third-party skin grafts (n=3) were rejected. Prevention of rat-lung allograft rejection could be achieved by intravenous administration of CTLA4Ig, resulting in long-term allograft survival with acceptance of donor-specific skin grafts. PMID- 12096785 TI - Defining the role of a tailored luminal solution for small bowel preservation. AB - The mucosal layer is the initial site of small bowel (SB) graft injury sustained during cold storage. Vascular administration of preservation solutions alone is unable to prevent ischemic injury of this layer during clinically relevant storage periods. The SB is unique in that it possesses both a vascular and a luminal route by which preservation solutions can be administered. We hypothesized that addition of a luminal-delivered solution, formulated on amino acid requirements for energy- and non-energy-related reactions, would provide site-specific preservation of mucosal energetics, barrier function and morphology throughout an extended period of cold storage. Of the three luminal solutions containing amino acids which were tested (UWG, AA1, AA2), only the two groups (AA1, AA2), containing glutamine plus 18 other amino acids, +/- osmotic agent (lactobionate) and buffer (BES), exhibited significant improvements in energetics, barrier function, and histology compared to the clinical standard of isolated vascular University of Wisconsin (UW) solution. Although the AA1 and AA2 groups preserved barrier function and morphology up to 24h better than all other solutions tested, AA2 proved to be the only luminal solution with values of permeability, conductance, and short-circuit current not significantly different from freshly isolated tissues. Furthermore, the greatest reduction in histologic injury was effected by AA2 treatment (median grade 2 compared to control, UW(v), grade 8). This study documents that a luminal-delivered solution, formulated on physiologic SB requirements, provides targeted preservation of the SB mucosa. PMID- 12096786 TI - Genetically modified adenovirus vector containing an RGD peptide in the HI loop of the fiber knob improves gene transfer to nonhuman primate isolated pancreatic islets. AB - The ability to transfer immunoregulatory, cytoprotective, or antiapoptotic genes into pancreatic islets (PIs) may allow enhanced post-transplantation survival. The available gene transfer vectors differ greatly in their ability to infect and express genes in different cell types. One limitation associated with the use of viral vectors is related to the virus reliance on the presence of its primary binding site. Tropism of the viral vectors can be altered using retargeting strategies. Results on phage biopanning proved that the RGD motif has in vivo targeting capabilities. This motif interacts especially with cellular integrins of the alphavbeta3 and alphavbeta5 types, highly expressed on pancreatic islets. In this report, we have explored the utility of a retargeted adenovirus vector (Ad) containing an RGD motif in the HI loop of the fiber knob in order to improve the infection efficiency to intact isolated nonhuman primate PIs and reduce toxicity after the genetic modification. Nonhuman primate Pis were isolated by a semi-automated technique. Steptozotocin-induced diabetic mice with severe combined immunodeficiency disease (SCID) were used as recipients. A recombinant Ad containing a heterologous RGD peptide and expressing luciferase (AdRGDLuc) or green fluorescent protein (AdRGDGFP) were generated in our laboratory. Similar Ads without the RGD peptide were used as a control (AdLuc and AdGFP). Higher transfection efficiency was demonstrated using AdRGDGFP compared with AdGFP (>80% of the islet cells were infected at 10 particle-forming units (pfu)/cell using AdRGDGFP vs. 7% after infection with AdGFP).More than 90% of the infected cells were insulin-producing cells. Significantly higher transgene expression was demonstrated after infection with AdRGDLuc compared with AdLuc at different titers. Analysis of the glucose-stimulated insulin response demonstrated better performance of PI transfected with AdRGDLuc at low titers (10 pfu/cell in order to achieve > 80% transfection efficiency) compared with AdLuc at high titers. Finally, long-term euglycemia (>250d) was observed in 89% of the animals that received PI infected with AdRGDLuc compared with none of the animals that received PI infected with AdLuc. The present study provides new information about the possibility of tropism modification of Ad vectors to increase the transfection efficiency and transgene expression to isolated PI. Incorporation of the RGD sequence in the HI loop of the fiber knob allows highly efficient transfection efficiency to nonhuman primate insulin-producing cells and adequate long-term function of the p-cell after transplantation. PMID- 12096787 TI - Hyperhomocyst(e)inemia induces accelerated transplant vascular sclerosis in syngeneic and allogeneic rat cardiac transplants. AB - Chronic rejection (CR) and transplant vascular sclerosis (TVS) cause the majority of graft failures in cardiac transplantation. Hyperhomocyst(e)inemia [hH(e)] is associated with human TVS without a proven causal relationship. This study investigated the effect of hH(e) on graft survival and TVS in allogeneic and syngeneic rat cardiac transplants. Lewis recipients of heterotopic F344 heart allografts, received normal or hH(e)-inducing (folate, methionine) diets [controls: syngeneic transplanted [+/- hH(e), + CsA] and nontransplanted rats [+/ hH(e), +/- CsA]]. Serial plasma homocyst(e)ine [H(e)] levels were measured. TVS was assessed in clinically rejected grafts and a subset of pre-rejection normal diet allografts (day 64) (neointimal index, NI). The hH(e) diet elevated plasma H(e) levels. When compared with normal diet controls (n = 9), hH(e) diet allografts (n = 9) had decreased time to onset of CR (40 +/- 9 vs. 72 +/- 10d, p = 0.02), and graft failure (64 +/- 10 vs. 107 +/- 12d, p = 0.009). hH(e) diet allografts at rejection (n = 9, 64d) had more severe TVS (NI = 68 +/- 2) than both time-matched normal diet allografts (NI = 49 +/- 6, n = 8, 64d, p <0.001) and normal diet allografts at rejection (NI = 58 +/- 5, n = 9, 107d, p = 0.007). hH(e) induced TVS in syngeneic grafts (NI=50 +/- 3, n = 10 vs. NI = 5 +/- 3, n = 10, 130d, p <0.001). hH(e) accelerated rejection and increased the severity of TVS in allogeneic cardiac transplants, and induced TVS in syngeneic cardiac transplants. PMID- 12096788 TI - Can a transplanted living donor kidney function equivalently to its native partner? AB - Early renal functional adaptation was examined in 81 haploidentical donor and recipient pairs, as well as long-term stability of glomerular filtration rate (GFR) in 78 recipients. GFR was determined pre- and 1 month postnephrectomy in donors and 1 month post-transplant and yearly thereafter in recipients. Compensatory increase in filtration (CIF) of transplanted and native kidneys was calculated using donor pretransplant GFR: [CIF= (GFR at 1 month/donor prenephrectomy GFR) x 100]. Annual rates of change in GFR were estimated using within-patient linear regression analysis (slopes). Recipients without rejection (n = 62) and their donors had similar early GFR and CIF. Those with acute rejection (n = 19) had significantly lower GFR and CIF than their donors (61 +/- 16 mL/min/1.73 m2 and 57 +/- 14% vs. 75 +/- 11 and 69 +/- 9; p = 0.01 and p < 0.001). Recipients without cyclosporine (n = 52) had 1 month GFR and CIF of 70 +/ 14 and 67 +/- 14 vs. 72 +/- 11 and 69 +/- 11 for their donors. Those with cyclosporine (n = 29) had 1 month GFR and CIF of 64 +/- 14 and 62 +/- 16 vs. 69 +/- 12 and 67 +/- 11 for their donors (p = 0.15 and 0.16). Comparison of median (25th, 75th) rates of change of GFR with and without acute rejection or cyclosporine did not demonstrate significant effects of either on stability of allograft function, although there was a trend towards greater loss of GFR in cyclosporine-treated patients [-1.1 (-2.5, 0.8) vs. 0.0 (-1.8, 1.2) mL/min/1.73 m2/year, p = 0.47]. We conclude that, in the absence of rejection, the transplanted kidney maintains the same capacity for functional adaptation as its native partner. Therapy with cyclosporine does not significantly inhibit early physiological adaptation of renal transplants. PMID- 12096789 TI - First clinical use of a novel bioartificial liver support system (BLSS). AB - The first clinical use of the Excorp Medical Bioartificial Liver Support System (BLSS) in support of a 41-year-old African-American female with fulminant hepatic failure is described. The BLSS is currently in a Phase I/II safety evaluation at the University of Pittsburgh/UPMC System. Inclusion criteria for the study are patients with acute liver failure, any etiology, presenting with encephalopathy deteriorating beyond Parson's Grade 2. The BLSS consists of a blood pump; a heat exchanger to control blood temperature; an oxygenator to control oxygenation and pH; a bioreactor; and associated pressure and flow alarm systems. Patient liver support is provided by 70-100 g of porcine liver cells housed in the hollow fiber bioreactor. The patient exhibited transient hypotension and thrombocytopenia at initiation of perfusion. The only unanticipated safety event was a lowering of patient glucose level at the onset of perfusion with the BLSS that was treatable with intravenous glucose administration. Moderate changes in blood biochemistries pre- and post perfusion are indicative of liver support being provided by the BLSS. While the initial experience with the BLSS is encouraging, completion of the Phase I/II study is required in order to more fully understand the safety aspects of the BLSS. PMID- 12096790 TI - Sensitisation to swine leukocyte antigens in patients with broadly reactive HLA specific antibodies. AB - We have previously shown that IgG HLA specific antibodies in the sera of highly sensitised patients awaiting renal transplantation can cross-react with swine leukocyte antigens (SLA). In this study we determined the frequency of patient serum IgG HLA specific antibody binding to a porcine lymphocyte panel and the likelihood of locating a cross-match negative pig donor for sensitised patients. Serum samples (n = 82) were obtained from 35 sensitised [current IgG panel reactive antibodies (PRA) > 10%] and seven nonsensitised patients awaiting renal transplantation at Addenbrooke's Hospital, Cambridge, UK. Fifty sera had IgG HLA specific PRA of 11-84%, 20 had IgG PRA of >84% and 12 had 0% PRA (negative controls). Sera were absorbed with porcine erythrocytes to remove xenoreactive natural antibodies and tested for cross-reactive IgG HLA specific antibody binding by flow cytometry against a panel of porcine lymphocytes obtained from 23 human decay accelerating factor (hDAF) transgenic pigs. A total of 1,884 cross match combinations were tested and 369 (20%) gave a positive porcine lymphocyte cross-match. For sera from sensitised patients with IgG PRA (11-64%), only 6 of 805 (0.75%) cross-match tests were positive. In contrast, for sera from patients with high IgG PRA (>64%), 363 of 805 (45%) cross-match tests were positive (p < 0.0001). There was no difference in the frequency of positive cross-matches between patient sera with IgG PRA 65-84% and highly sensitised patient sera with IgG PRA 85-100% [156/345 (45%) vs. 207/460 (45%)]. This study demonstrates that only patient sera with broadly reactive IgG HLA specific PRA (>64%) cross-react with porcine lymphocytes. If future clinical trials of xenotransplantation are undertaken, it may be of value to select a cross-match-negative pig organ donor for such patients. PMID- 12096791 TI - Acute coronary syndromes after renal transplantation in patients with end-stage renal disease resulting from diabetes. AB - Coronary heart disease is the leading cause of death in both diabetes mellitus and end-stage renal disease. Although renal transplantation is known to reduce mortality in end-stage renal disease, its effect on the incidence of acute coronary syndromes is unknown. Using data from the United States Renal Data System, we studied 11,369 patients with end-stage renal disease due to diabetes enrolled on the renal and renal-pancreas transplant waiting list from 1 July 1994 to 30 June 1997. Cox nonproportional hazards regression models were used to calculate the adjusted, time-dependent relative risk for the most recent hospitalization for acute coronary syndromes (including acute myocardial infarction, unstable angina, or other acute coronary syndromes, ICD9 Code 410.x or 411.x) for a given patient in the study period. Demographics and comorbidities were controlled by using data from the medical evidence form (HCFA 2728). After renal transplantation, patients had an incidence of acute coronary syndromes of 0.79% per patient year, compared to 1.67% per patient year prior to transplantation. In comparison to maintenance dialysis, renal transplantation was independently associated with a lower risk for acute coronary syndromes (hazard ratio 0.38, 95% confidence interval, 0.30-0.49). Patients with end-stage renal disease due to diabetes on the renal transplant waiting list were much less likely to be hospitalized for acute coronary syndromes after renal transplantation. The reasons for this decreased risk should be the subject of further study. PMID- 12096792 TI - Declining influence of race on the outcome of living-donor renal transplantation. AB - A racial disparity in graft survival for renal transplant recipients has been documented for both cadaveric and living-donor transplants. In the present single center study we analyzed graft survival by race for recipients of living-donor kidney transplants in three eras: 1985-89, 1990-94, and 1995-2000. There was an intensification of the immunosuppressant regimen beginning in 1996, such that all patients received cyclosporine or tacrolimus with mycophenolate and prednisone. Graft survival was analyzed using the Cox proportional hazards model. There were 79 black recipients and 210 white recipients with no difference in mean age, degree of HLA matching, or proportion of recipients with diabetes as the cause of end-stage renal disease. Using all data from 1985 to 2000, graft survival was significantly better for whites vs. blacks adjusted for age, gender, diabetes, era of the transplant, and haplotype match (p = 0.05). However, when analyzed by era, there was a temporal trend for a progressive decrease in the racial disparity in graft survival. In confirmation of this effect, there was a significant race-era interaction (p = 0.01) on multivariable Cox proportional hazards analysis. The most recent data from the United States Renal Data System (USRDS) show a similar decrease in the racial difference in 1-year graft survival. We conclude that the influence of race on living-donor graft survival is diminishing over time. PMID- 12096793 TI - Influenza virus infection in adult solid organ transplant recipients. AB - BACKGROUND: Solid organ transplant (SOT) recipients have been reported to be more susceptible to influenza virus. However, little is known about the clinical epidemiology and the implications of influenza viral infection among SOT recipients. METHODS: Cohort study of influenza viral infection in SOT recipients at the University of Pittsburgh Medical Center. RESULTS: Between November 1990 and April 2000, 30 cases of influenza were diagnosed in SOT recipients at our center, including influenza A (n = 22) and influenza B (n = 8). These included recipients of lung (n = 19), liver (n = 5) and kidney (n = 6) transplants. The incidence of influenza viral infection was 41.8 cases/1,000 person years (PYs), 2.8 cases/1000 PYs and 4.3 cases/ 1,000 PYs among lung, liver and renal transplant patients, respectively (p <0.0001). Symptoms were reported in all patients and included malaise, myalgia/ arthralgia, fever, cough, and shortness of breath. Secondary bacterial pneumonia occurred in five patients (17%). Other complications were seen in three SOT recipients (2 liver and 1 kidney) and included: myocarditis, myositis, and bronchiolitis obliterans. Biopsy of the transplanted organ was performed in 21 SOT recipients (18 lung, 1 liver and 2 kidney) at the time of influenza viral infection. Overall, 62% (13/21) showed variable degrees of acute allograft rejection, and included 61% (11/18) of lung, and 100% (2/2) of kidney transplant recipients. CONCLUSIONS: Influenza infection is associated with significant morbidity in different groups of SOT recipients. Studies are needed to determine if yearly chemoprophylaxis with antiviral drugs might benefit this patient population. PMID- 12096794 TI - Duration of donor brain death and its influence on kidney graft function. AB - Short- and long-term rates of success after cadaveric kidney transplantation are significantly inferior to those from living related or unrelated donors. The major difference between cadaveric and living donation is brain death. In the present study we analyzed the influence of duration of brain death on short- and long-term graft function after cadaveric kidney transplantation. The interval between declaration of donor brain death and the beginning of the cold ischemia time before graft explantation was defined as duration of brain death (DBD). The influence of DBD on incidence of primary graft function and on duration of delayed kidney graft function as well as on kidney graft survival was analyzed in 1106 patients transplanted in one center and confirmed in a validation study of a second series of 752 kidney graft recipients from another transplant center. Kidney grafts harvested from donors with longer DBD (>470 min) exhibited a significantly higher incidence of primary graft function and a significantly better graft survival rate in comparison to kidneys from donors with a shorter DBD (<470 min). The tendency of these results could be confirmed in an independent validation study; however, the differences were not statistically significant. Although the dramatic hemodynamic and immunological changes in brain dead donors may impair the quality of a potential kidney transplant, a longer duration of donor brain death did not deteriorate early and long-term kidney graft function. PMID- 12096795 TI - Genetic dissection of the initiation of the infection process and nodule tissue development in the Rhizobium-pea (Pisum sativum L.) symbiosis. AB - Twelve non-nodulating pea (Pisum sativum L.) mutants were studied to identify the blocks in nodule tissue development. In nine, the reason for the lack of infection thread (IT) development was studied; this had been characterized previously in the other three mutants. With respect to IT development, mutants in gene sym7 are interrupted at the stage of colonization of the pocket in the curled root hair (Crh- phenotype), mutants in genes sym37 and sym38 are blocked at the stage of IT growth in the root hair cell (Ith- phenotype) and mutants in gene sym34 at the stage of IT growth inside root cortex cells (Itr- phenotype). With respect to nodule tissue development, mutants in genes sym7, sym14 and sym35 were shown to be blocked at the stage of cortical cell divisions (Ccd- phenotype), mutants in gene sym34 are halted at the stage of nodule primordium (NP) development (Npd- phenotype) and mutants in genes sym37 and sym38 are arrested at the stage of nodule meristem development (Nmd- phenotype). Thus, the sequential functioning of the genes Sym37, Sym38 and the gene Sym34 apparently differs in the infection process and during nodule tissue development. Based on these data, a scheme is suggested for the sequential functioning of early pea symbiotic genes in the two developmental processes: infection and nodule tissue formation. PMID- 12096796 TI - X-ray microanalytical studies of mineral localization in the needles of white pine (Pinus strobus L.). AB - Eastern white pine (Pinus strobus L.) shoots from mature trees were collected from two sites of contrasting soil pH: the Glendon campus of York University in Toronto, Ontario (pH 6.7 at 40 cm); and Muskoka near Huntsville, Ontario (pH 4.2 at 40 cm). Needles of ages 1-3 years were removed from the shoots, and the percentage of ash and silica was determined for all ages. Other needles were frozen in liquid nitrogen and kept in a cryo-biological storage system before x ray microanalysis. Percentages of ash and silica were higher in the needles from Muskoka. Ash and silica increased with needle age for trees from the Muskoka site, but less so at the Toronto site. Of the 13 elements (Na, Mg, Al, Si, P, S, Cl, K, Ca, Mn, Fe, Cu and Zn) detected by microanalysis, Mn, Fe, Cu and Zn were detected in small amounts in the epidermis, endodermis and transfusion tissue (the layer of tracheids and parenchyma immediately surrounding the vascular bundles), and K, P, S and Cl were almost ubiquitous in distribution. Sodium was occasionally detected in the transfusion tissue, and magnesium was concentrated in the endodermal cells. The epidermal walls, transfusion tissue and endodermis were major sites of calcium localization. Silicon was concentrated in the extreme tips of the needles in all tissues, but particularly in the transfusion tissue, and more so in the Muskoka samples. Microanalysis revealed a higher Al content in the Muskoka needles, that Al was concentrated in the needle tips and that the transfusion tissues were major sites of accumulation. PMID- 12096797 TI - Does ethylene treatment mimic the effects of pollination on floral lifespan and attractiveness? AB - In some species pollination may result in rapid changes in perianth colour and form (petal senescence and abscission, flower closure), rendering the flowers less attractive to pollinators. It has been suggested that this effect is mediated by ethylene. Flowers from about 200 species and 50 families were exposed to ethylene (3 ppm for 24 h at 20 degrees C). The effects on petal senescence and abscission have been described previously. Flower closure and perianth colour changes were generally ethylene-sensitive, but responses showed no consistency within families. Several flowers known to respond to pollination by rapid cessation of attractiveness were also exposed to ethylene: this produced the same effect as pollination, both on flower colour and form. Species that respond to pollination by changing flower form or colour were found exclusively in families in which the species are generally ethylene-sensitive (with regard to changes in perianth form and colour). However, several families are generally ethylene sensitive but contain no species reported to respond to pollination. PMID- 12096798 TI - Consequences of stoichiometric error on nuclear DNA content evaluation in Coffea liberica var. dewevrei using DAPI and propidium iodide. AB - The genome size of coffee trees (Coffea sp.) was assessed using flow cytometry. Nuclear DNA was stained with two dyes [4',6-diamino-2-phenylindole dihydrochloride hydrate (DAPI) and propidium iodide (PI)]. Fluorescence in coffee tree nuclei (C-PI or C-DAPI) was compared with that of the standard, petunia (P PI or P-DAPI). If there is no stoichiometric error, then the ratio between fluorescence of the target nuclei and that of the standard nuclei (R-PI or R DAPI) is expected to be proportional to the genome size. Between-tree differences in target : standard fluorescence ratios were noted in Coffea liberica var. dewevrei using propidium iodide and DAPI. For both dyes, between-tree differences were due to a lack of proportionality when comparing locations of the coffee peak and the petunia peak. Intraspecific genome size variations clearly cannot explain variations in the target : standard fluorescence ratio. The origin of the lack of proportionality between target and standard fluorescences differed for the two dyes. With propidium iodide, there was a regression line convergence point, and no between-tree differences were noted in this respect, whereas there was no such convergence with DAPI. An accurate estimate of genome size can thus be obtained with PI. Implications with respect to accessibility and binding mode are discussed. PMID- 12096800 TI - Allometric relationships to estimate seasonal above-ground vegetative and reproductive biomass of Vitis vinifera L. AB - A procedure is described for obtaining allometric regression equations to estimate non-destructively and in a cost-effective manner the current year's above-ground vegetative and reproductive biomass of Vitis vinifera L. Merlot' throughout the growing season. Significant relationships were obtained over a 3 year period (1998-2000) between the dimensions of an individual shoot per vine (i.e. diameter and length) and dry weights of its primary stem, primary leaves and lateral growth. The dry mass of a grape was best estimated from measurements of the basal diameter of the bunch peduncle. Introducing cumulative degree-days as an additional explanatory variable in the equations allowed them to be used irrespective of year and growth stage. Multi-year regressions were used to quantify in detail the seasonal evolution of mature grapevine biomass under the climatic conditions of the Bordeaux area, France, and for differing levels of soil nitrogen. PMID- 12096799 TI - In vivo characterization of the effects of abscisic acid and drying protocols associated with the acquisition of desiccation tolerance in alfalfa (Medicago sativa L.) somatic embryos. AB - Although somatic embryos of alfalfa (Medicago sativa L.) had acquired some tolerance to desiccation at the cotyledonary stage of development (22 d after plating), additional culturing in 20 microm abscisic acid (ABA) for 8 d induced greater desiccation tolerance, as determined by increased germination. Compared with fast drying, slow drying of the ABA-treated embryos improved desiccation tolerance. However, slow drying of non-ABA-treated embryos led to the complete loss of germination capacity, while some fast-dried embryos survived. An electron paramagnetic resonance spin probe technique and in vivo Fourier transform infrared microspectroscopy revealed that cellular membrane integrity and a helical protein secondary structure were maintained during drying in embryos cultured in media enriched with 20 microM ABA, but not in embryos cultured in the absence of ABA. Slow-dried, non-ABA-treated embryos had low oligosaccharide to sucrose ratios, an increased proportion of beta-sheet protein secondary structures and broad membrane phase transitions extending over a temperature range of more than 60 degrees C, suggestive of irreversible phase separations. The spin probe study showed evidence of imbibitional damage, which could be alleviated by prehydration in humid air. These observations emphasize the importance of appropriate drying and prehydration protocols for the survival and storage of somatic embryos. It is suggested that ABA also plays a role in suppressing metabolism, thus increasing the level of desiccation tolerance; this is particularly evident under stressful conditions such as slow drying. PMID- 12096801 TI - Maintaining apical dominance in the fern gametophyte. AB - A kinetic model is developed for cell differentiation in the fern gametophyte to test hypotheses on the role of spatially patterned plasmodesmata networks in development. Of particular interest is the establishment and maintenance of apical cell type in a single cell, with concurrent suppression of this character in all other cells (apical dominance). Steps towards understanding apical cell localization in geometrically simple gametophytes may shed light on the establishment and maintenance of apical meristems in higher plants. The model, based on the plasmodesmata maps of Tilney and colleagues and involving kinetics for a requisite minimum of two morphogens. successfully produces the apical/non apical cell differentiation patterns of normal development, and redifferentiation due to cell isolation, in six stages from 0-30 d of development. Our results indicate that increasing apical cell plasmodesmata number, as development progresses, is not required for effective transport across apical cell walls in maintaining apical dominance. PMID- 12096802 TI - In vitro bulb development in shallot (Allium cepa L. Aggregatum Group): effects of anti-gibberellins, sucrose and light. AB - Bulbing was studied in shallot plants cultured in vitro. Bulbing occurred under a 16 h photoperiod with fluorescent + incandescent light and 30-50 g 1(-1) sucrose in the culture medium. Exogenous gibberellin (10 microM GA3) inhibited leaf and root growth and bulbing. When added to the medium at a concentration of 10 microM, three inhibitors of gibberellin biosynthesis (ancymidol, flurprimidol and paclobutrazol) promoted bulb formation and the percentage of bulbing. When ancymidol was used in combination with GA3, it did not reverse the effect of GA3 applied alone. Under treatments with 30-70 g l(-1) sucrose, bulbing ratios greater than those found in control plants were achieved by addition of ancymidol, and bulb fresh weight was increased in the same way. Ancymidol caused a 66% decrease in sucrose content in leaf bases but greatly increased the glucose, fructose and fructan contents. The increase in fructan content by ancymidol could result from the three-fold rise in total [14C]sucrose uptake per plant from the culture medium associated with a marked increase in leaf base labelling at the expense of root labelling. The possible role of ancymidol is discussed and evidence supports a major regulatory role for gibberellins in bulbing. PMID- 12096803 TI - Temporal and spatial patterns of seed dispersal in two Cistus species (Cistaceae). AB - Cistus species are obligate seeding, early colonizers that follow disturbance, particularly fire, in Mediterranean ecosystems. We studied seed release, seed dispersal and soil seed populations in stands of Cistus ladanifer and C. libanotis. Seed release started in mid- to late summer (C. ladanifer) or in early autumn (C libanotis), and continued for a very extended period: 8-10 months in C. ladanifer, and for a mean of 16 months in C. libanotis. The xerochastic capsules of both species released seeds by successive dehiscence of the locules. All capsules begin to dehisce simultaneously at the start of the seed release period, but in C. libanotis capsule fragmentation replaced dehiscence early in the seed release period. In plants of both species, seed shadows were characterized by a peak of density beneath the plant canopy and a very short tail of much lower densities, indicating that seeds are concentrated beneath mother plants when dispersed. Nevertheless, in late May, at the onset of the fire season, soil seed densities beneath plant canopies were low compared with densities expected from seed shadows, but were apparently high enough to allow recovery of the stands if a disturbance, such as fire, had taken place. Seed-eating Bruchidae in summer and granivorous ants during the seed release period were apparently the main causes of seed losses. Results suggest that in both Cistus species, the staggered seed release could constitute an efficient risk-reducing trait. The plant pool of seeds existing throughout most of the year could be a relevant component of Cistus seed banks. PMID- 12096804 TI - The effect of nitrogen nutrition on cluster root formation and proton extrusion by Lupinus albus. AB - Nitrogen nutrition can influence cluster root formation in many wild species, but the effect of N form on cluster root formation and root exudation by white lupin is not known. In a solution culture study, we examined the effect of N nutrition (ammonium, nitrate, both or N2 fixation) on cluster root formation and H+ extrusion by white lupin plants under deficient and adequate P supply. The number of cluster roots increased greatly when plants were supplied with I microM P compared with 50 microM P, the increase being 7.8-fold for plants treated with (NH4)2SO4, 3-fold for plants treated with KNO3 and NH4NO3, and 2-4-fold for N2 fixing plants. Under P deficiency. NH4+-N supply resulted in production of a greater number and biomass of cluster roots than other N sources. Dry weight of cluster roots was 30 % higher than that of non-cluster roots in P-deficient plants treated with (NH4)2SO4 and NH4NO3. In plants treated with sufficient P (50 microM), the weight of non-cluster roots was approx. 90 % greater than that of cluster roots. Both total (micromol per plant h(-1)) and specific (micromol g(-1) root d. wt h(-1)) H+ extrusions were greatest from roots of plants supplied with (NH4)2SO4, followed by those supplied with NH4NO3 and N2 fixation, whereas plants receiving KNO3 had negative net H+ extrusion between the third and fifth week of growth (indicating uptake of protons or release of OH- ions). The rate of proton extrusion by NH4+-N-fed plants was similar under P-deficient and P-sufficient conditions. In contrast, proton exudation by N2-fixing plants and KNO3-treated plants was ten-fold greater under P deficiency than under P sufficiency. In comparison with P deficiency, plants treated with 50 microM P had a significantly higher concentration of P in roots, shoots and youngest expanded leaves (YEL). Compared with the N2 fixation and KNO3 treatments, total N concentration was highest in roots, shoots and YEL of plants supplied with (NH4)2SO4 and NH4NO3, regardless of P supply. Under P deficiency, K concentrations in roots decreased at all N supplies, especially in plants treated with (NH4)2SO4 and NH4NO3, which coincided with the greatest H+ extrusion at these P and N supplies. In conclusion, NH4-N nutrition stimulated cluster root formation and H+ extrusion by roots of P-deficient white lupin. PMID- 12096805 TI - Aluminium and phosphate uptake by Phragmites australis: the role of Fe, Mn and Al root plaques. AB - Aluminium, a potentially phytotoxic metal, is an important constituent of many mine water discharges but has largely been neglected in the literature. The behaviour of this element in the rhizosphere of the wetland plant Phragmites australis was investigated in the laboratory in the presence and absence of Mn and Fe root plaques. Electron microscopy and chemical extraction techniques were utilized to determine the physico-chemical properties of the plaques and any association of Al. Both Mn and Fe plaques occurred as amorphous coatings on root surfaces with uneven distributions. Al was not adsorbed onto the surface of either plaque type but formed a separate phosphate deposit closely resembling the Fe and Mn plaques. Phosphorus was also found to be adsorbed to the surface of the Fe plaques (but not the Mn plaques). Both mechanisms were found to immobilize P at the root surface but this did not significantly reduce the concentration of P in aerial plant tissues that was sufficient to ensure adequate growth. PMID- 12096806 TI - Instantaneous canopy photosynthesis: analytical expressions for sun and shade leaves based on exponential light decay down the canopy and an acclimated non rectangular hyperbola for leaf photosynthesis. AB - Analytical expressions for the contributions of sun and shade leaves to instantaneous canopy photosynthesis are derived. The analysis is based on four assumptions. First, that the canopy is closed in the sense that it is horizontally uniform. Secondly, that there is an exponential profile of light down the canopy with the same decay constant for light from different parts of the sky. Thirdly, that the leaf photosynthetic response to incident irradiance can be described by a three-parameter non-rectangular hyperbola (NRH). And lastly, that light acclimation at the leaf level occurs in only one parameter of the NRH, that describing the light-saturated photosynthetic rate, which is assumed to be proportional to the local averaged leaf irradiance. These assumptions have been extensively researched empirically and theoretically and their limitations are quite well understood. They have been widely used when appropriate. Combining these four assumptions permits the derivation of algebraic expressions for instantaneous canopy photosynthesis which are computationally efficient because they avoid the necessity for numerical integration down the canopy. These are valuable for modelling plant and crop ecosystems, for which canopy photosynthesis is the primary driver. Ignoring the sun/shade dichotomy can result in overestimates of canopy photosynthesis of up to 20 %, but using a rectangular hyperbola instead of a non-rectangular hyperbola to estimate canopy photosynthesis taking account of sun and shade leaves can lead to a similarly sized underestimate. PMID- 12096807 TI - Glandular hairs of Sigesbeckia jorullensis Kunth (Asteraceae): morphology, histochemistry and composition of essential oil. AB - Long-stalked glandular hairs of outer and inner involucral bracts of Sigesbeckia jorullensis, which are important for epizoic fruit propagation, were investigated using light and scanning electron microscopy. The essential oil secreted by the hairs was analysed by chromatographic methods including gas chromatography/mass spectrometry and with a laser microprobe mass analyser. The glandular hairs consisted of a large multicellular stalk and a multicellular secreting head. The apical layer of glandular head cells was characterized by leucoplasts and calcium oxalate crystals. Below the apical cells there were up to six layers of cells containing many chloroplasts around the nucleus and surrounded by vacuoles filled with flavonoids and tannins. The essential oil originating in the head cells was secreted into the subcuticular space and may be liberated by rupture of the cuticle. It was mainly composed of sesqui- and diterpenes, with the sesquiterpene hydrocarbon germacrene-D as the main component. Monoterpenes, n-alkanes and their derivatives as well as flavonoid aglycones were also detected. The stickiness of the essential oil is probably associated with the high content of oxygenated sesqui- and diterpenes. In addition to long-stalked trichomes, small biseriate trichomes occurred, secreting small quantities of essential oil into a subcuticular space. PMID- 12096808 TI - Quantitative analysis of shoot development and branching patterns in Actinidia. AB - We developed a framework for the quantitative description of Actinidia vine architecture, classifying shoots into three types (short, medium and long) corresponding to the modes of node number distribution and the presence/ absence of neoformed nodes. Short and medium shoots were self-terminated and had only preformed nodes. Based on the cut-off point between their two modes of node number distribution, short shoots were defined as having nine or less nodes, and medium shoots as having more than nine nodes. Long shoots were non-terminated and had a number of neoformed nodes; the total number of nodes per shoot was up to 90. Branching patterns for each parent shoot type were represented by a succession of branching zones. Probabilities of different types of axillary production (latent bud, short, medium or long shoot) and the distributions of length for each branching zone were estimated from experimental data using hidden semi-Markov chain stochastic models. Branching was acrotonic on short and medium parent shoots, with most axillary shoots being located near the shoot tip. For long parent shoots, branching was mesotonic, with most long axillary shoots being located in the transition zone between the preformed and neoformed part of the parent shoot. Although the shoot classification is based on node number distribution there was a marked difference in average (per shoot) internode length between the shoot types, with mean values of 9, 27 and 47 mm for short, medium and long shoots, respectively. Bud and shoot development is discussed in terms of environmental controls. PMID- 12096809 TI - Molecular cytogenetic analysis of Podocarpus and comparison with other gymnosperm species. AB - DNA sequences have been mapped to the chromosomes of Podocarpus species from New Zealand and Australia by fluorescent in situ hybridization. Unlike other conifers, these species show only one pair of major sites of 45S rDNA genes, and two additional minor sites were seen in the Australian P. lawrencei. Unusually, 45S sequences collocalize to the same chromosomal region as the 5S rDNA. The telomere probe (TTTAGGG)n hybridizes to the ends of all chromosomes as well as to a large number of small sites distributed along the length of all chromosomes. Two other simple sequence repeats, (AAC)5 and (GATA)4, show a diffuse pattern of hybridization sites distributed along chromosomes. Southern blots using a variety of probes obtained from the reverse transcriptase of retroelements (gypsy, copia and LINE) from P. totara, P. nivalis and Dacrycarpus dacrydioides show that these retroelements are abundant and widespread in Podocarpaceae and also in others conifers. Some retroelements such as copia pPonty3 and gypsy pPot1li are more abundant in the genome of Picea abies and Ginkgo biloba than in the species from which they were amplified. PMID- 12096811 TI - An ecological interpretation of the difference in leaf anatomy and its plasticity in contrasting tree species in orange kloof, table mountain, South Africa. AB - Leaf anatomy and morphology were studied in 11 tree species growing in an undisturbed forest and the adjoining fynbos for over 50 years. Functional anatomical results suggest that the forest and the fynbos are ecologically distinct. Moreover, leaf anatomy suggests that the foliage is primarily adapted for photosynthesis rather than for control of transpirational water loss. Forest precursor tree species and scrub species exhibit xeromorphy in the fynbos whereas they exhibit mesomorphic features inside the forest. The wide-ranging species, such as Olea capensis subsp. capensis, simulated the response of the forest precursors, with the cuticle being phenotypically plastic between the forest and the fynbos but not between the stream and non-stream habitats. Finally, the forest precursors, the scrub species, and the wide-ranging taxa seem to have anatomical characters which can be modified in the fynbos and therefore allow its colonization by a variety of different species. PMID- 12096810 TI - Green fluorescent protein as a visual marker in somatic hybridization. AB - Using a transgenic citrus plant expressing Green Fluorescent Protein (GFP) as a parent in somatic fusion experiments, we investigated the suitability of GFP as an in vivo marker to follow the processes of protoplast fusion, regeneration and selection of hybrid plants. A high level of GFP expression was detected in transgenic citrus protoplasts, hybrid callus, embryos and plants. It is demonstrated that GFP can be used for the continuous monitoring of the fusion process, localization of hybrid colonies and callus, and selection of somatic hybrid embryos and plants. PMID- 12096813 TI - Periods of organogenesis in shoots of Nothofagus dombeyi (Mirb.) oersted (Nothofagaceae). AB - The organogenetic cycle of main-branch shoots of Nothofagus dombeyi (Nothofagaceae) was studied. Twelve samples of 52-59 parent shoots were collected from a roadside population between September 1999 and October 2000. Variations over time in the number of nodes of terminal and axillary buds, and the length, diameter and number of leaves of shoots derived from these buds (sibling shoots) were analysed. The number of nodes of buds developed by parent shoots was compared with the number of nodes of buds developed, I year later, by sibling shoots. The length, diameter and number of leaves of sibling shoots increased from October 1999 to February 2000 in those shoots with a terminal bud. However, extension of most sibling shoots, including the first five most distal leaf primordia, ceased before February due to abscission of the shoot apex. Axillary buds located most distally on a shoot had more nodes than both terminal buds and more proximal axillary buds. The longest shoots included a preformed part and a neoformed part. The organogenetic event which initiated the neoformed organs continued until early autumn, giving rise to the following year's preformation. The absence of cataphylls in terminal buds could indicate a low intensity of shoot rest. The naked terminal bud of Nothofagus spp. could be interpreted as a structure less specialized than the scaled bud found in genera of Fagaceae and Betulaceae. PMID- 12096812 TI - Effects of a stay-green mutation on plant nitrogen relations in Lolium perenne during N starvation and after defoliation. AB - The stay-green mutation of the nuclear gene sid results in inhibition of chlorophyll degradation during leaf senescence in grasses, reducing N remobilization from senescing leaves. Effects on growth of Lolium perenne L. were investigated during N starvation (over 18 d) and after severe defoliation, when leaf growth depends on the remobilization of internal N. Rates of dry mater production, partitioning between shoots and roots, and re-partitioning of N from shoots to roots were very similar in stay-green and normal plants under N starvation. Km and Vmax for net uptake of NH4+ were also similar for both genotypes, and Vmax increased with the duration of N deprivation. The mutation had little effect on recovery of leaf growth following severe defoliation, but stay-green plants recommenced NO3- and K+ uptake 1 d later than normal plants. Import of remobilized N into new leaves was generally similar in both lines. However, stay-green plants remobilized less N from stubble compared with normal plants. It was concluded that the sid locus stay-green mutation has no significant adverse effect on the growth of L perenne during N starvation, or recovery from severe defoliation when plants are grown under an optimal regime of NO3- supply both before and after defoliation. The absence of any effect on leaf dry matter production implies that the difference in foliar N availability attributable to this mutation has little bearing on productivity, at least in the short to medium term. PMID- 12096814 TI - Wild manihot species do not possess C4 photosynthesis. AB - Cultivated cassava (Manihot esculenta) has a higher rate of photosynthesis than is usual for C3 plants and photosynthesis is not light saturated. For these reasons it has been suggested that cultivated cassava could be derived from wild species possessing C4 photosynthesis. The natural abundance of 13C and activities of phosphoenolpyruvate carboxylase and phosphoglycolate phosphatase were measured in leaves of 20 wild cassava species to test this hypothesis. All the species studied, including M. flabellifolia the potential wild progenitor of cultivated cassava, clearly exhibited C3 not C4 characteristics. PMID- 12096815 TI - Structure and development of stomata on the primary root of Ceratonia siliqua L. AB - Stomata of various sizes are produced on the primary root of Ceratonia siliqua L. Most are generated during embryogenesis, prior to seed desiccation. They can be detected on the dry embryo in a wide zone just above the root tip. Initially, large stomata are formed. These have the ability to induce divisions of their neighbouring cells, creating particular cell patterns around them. Later, small perigenous stomata are generated. As the root grows following seed germination, the stomatal zone overlaps with that of the root hairs. Although root stomata of C. siliqua undergo a structural differentiation that seems almost identical to that of the elliptical stomata formed on leaves, they are unable to move and remain permanently open. Polarizing microscopy of fully differentiated stomata and young stomata at the stage of stomatal pore formation revealed deposition of radial cellulose microfibril systems on their periclinal walls. However, these systems were less developed than those on leaf stomata, a feature that might be responsible for their inactivity. Besides, plastids of the root guard cells (GCs) do not differentiate into chloroplasts but function solely as amyloplasts. Root stomata have a short life span. During rapid and intense root growth, GCs cannot keep pace with the elongation of their neighbouring rhizodermal cells. They therefore split in their mid-region, transversely to the stoma axis. The two parts of the transversely torn stoma are dragged apart and a large opening is formed on the root surface, just above the substomatal cavity. The root stomata, together with these openings, may facilitate increased gaseous exchange during respiration and/or an increased transfer of some nutrients and water in the rapidly growing primary root. PMID- 12096816 TI - Comparative genomics in the grass family: molecular characterization of grass genome structure and evolution. AB - The genomes of grasses are very different in terms of size, ploidy level and chromosome number. Despite these significant differences, it was found by comparative mapping that the linear order (colinearity) of genetic markers and genes is very well conserved between different grass genomes. The potential of such conservation has been exploited in several directions, e.g. in defining rice as a model genome for grasses and in designing better strategies for positional cloning in large genomes. Recently, the development of large insert libraries in species such as maize, rice, barley and diploid wheat has allowed the study of large stretches of DNA sequence and has provided insight into gene organization in grasses. It was found that genes are not distributed randomly along the chromosomes and that there are clusters of high gene density in species with large genomes. Comparative analysis performed at the DNA sequence level has demonstrated that colinearity between the grass genomes is retained at the molecular level (microcolinearity) in most cases. However, detailed analysis has also revealed a number of exceptions to microcolinearity, which have given insight into mechanisms that are involved in grass-genome evolution. In some cases, the use of rice as a model to support gene isolation from other grass genomes will be complicated by local rearrangements. In this Botanical Briefing, we present recent progress and future prospects of comparative genomics in grasses. PMID- 12096817 TI - Partial hybridization in wide crosses between cultivated sunflower and the perennial Helianthus species H. mollis and H. orgyalis. AB - To obtain introgressed sunflower lines with improved disease resistance, interspecific crosses were performed with foreign perennial species. We report on several unusual features displayed by these hybrid plants. The methods used to produce the kernels affected yield and genotypes of progeny. Phenotypic traits and DNA markers were investigated in 97 plants derived from cross-pollination between annual diploid cultivated sunflower (Helianthus annuus) and the perennial diploid species H. mollis or H. orgyalis, and the reverse reciprocal crosses. The level of hybridization in progeny was determined using RAPD and RFLP markers. Hybridization was performed by leaving embryos to develop normally on the head (classical crossing) or using embryo rescue. All observed plants derived from H. mollis were diploid (2n = 34). Phenotypes were predominantly similar to the female when cultivated sunflower was the female parent. Progeny from crosses using a wild species as the female parent resembled that parent. Thus, reciprocal crosses led to different progeny. F1 sister progeny shared different sets of molecular markers representing a few of those of the wild species used as the pollen donor. Our results indicate mechanisms leading to the unusual event of partial hybridization. Possible mechanisms behind these unusual events and their possible impact on evolution are discussed. PMID- 12096818 TI - Structure and function of shisham forests in central Himalaya, India: dry matter dynamics. AB - The biomass and net primary productivity (NPP) of 5- to 15-year-old Shisham (Dalbergia sissoo Roxb.) forests growing in central Himalaya were estimated. Allometric equations were developed for all above- and below-ground components of trees and shrubs for each stand. Understorey forest floor biomass and litter fall were also estimated in forest stands. The biomass (dry matter), forest floor biomass (standing crop litter), tree litter fall and NPP of trees and shrubs increased with increasing age of the forest stand, whereas the dry matter and herb NPP decreased significantly (P < 0.001) with increasing age of the forest. Total forest biomass and NPP ranged from 58.7 (5-year-old stand) to 136.1 t ha( 1) (15-year-old stand) and 12.6 (5-year-old stand) to 20.3 t ha(-1) year(-1) (15 year-old stand), respectively. Of these values, tree biomass accounted for 85.7 (5-year-old stand) to 90.1% (15-year-old) of total forest biomass, and tree NPP for 72.2 (5-year-old) to 82.3% (15-year-old) of total forest NPP. The biomass accumulation ratio (BAR) of the bole component (bole wood + bole bark) increased with increasing age of the forest stand. The bole BAR was 5.8 (5-year-old stand) to 7.9 (15-year-old stand). However, total BAR of the forest stand ranged from 5.5 (5-year-old) to 7.5 (15-year-old). PMID- 12096819 TI - Structure and function of shisham forests in central Himalaya, India: nutrient dynamics. AB - The structure and function of Shisham (Dalbergia sissoo Roxb.) forests were investigated in relation to nutrient dynamics in 5- to 15-year-old stands growing in central Himalaya. Nutrient concentrations and storage in different layers of vegetation were in the order: tree > shrub > herb. Forest soil, litter and vegetation accounted for 80.1-91.9, 1.0-1.5 and 7.0-18.4%, respectively, of the total nutrients in the system. There were considerable reductions (trees 32.8 43.1; shrubs 26.2-32.4; and herbs 18-8-22-2%) in nutrient concentrations of leaves during senescence. Nutrient uptake by the vegetation as a whole and also by the different components, with and without adjustment for internal recycling, was investigated. Annual transfer of litter nutrients to the soil from vegetation was 74.8-108.4 kg ha(-1) year(-1) N, 56.8-4 kg ha(-1) year(-1) P and 38.7-46.9 kg ha(-1) year(-1) K. Turnover rate and time for different nutrients ranged between 56 and 66 % year(-1) and 1.5 and 1.8 years, respectively. The turnover rate of litter indicates that over 50% of nutrients in litter on the forest floor are released, which ultimately enhances the productivity of the forest stand. The nutrient use efficiency in Shisham forests ranged from 136 to 143 kg ha(-1) year( 1) for N, 1,441 to 1,570 kg ha(-1) year(-1) for P and 305 to 311 kg ha(-1) year( 1) for K. Compared with natural oak forest (265 kg ha(-1) year(-1) and an exotic eucalypt plantation (18 kg ha(-1) year(-1), a higher proportion of nutrients was retranslocated in Shisham forests, largely because of higher leaf tissue nutrient concentrations. This indicates a lower nutrient use efficiency of Shisham compared with eucalypt and oak. Compartment models for nutrient dynamics have been developed to represent the distribution of nutrients pools and net annual fluxes within the system. PMID- 12096820 TI - Functional heterostyly in Tylosema esculentum (Caesalpinioideae). AB - Tylosema esculentum is a long-lived perennial species endemic to arid areas of southern Africa. Its potential as a crop species has long been recognized as a result of the high oil and protein content of its seeds. The reproductive biology and breeding systems of the species were investigated in wild and experimental populations growing in Botswana. Field observations confirmed that the species is heterostylous with the pistil and anthers exhibiting reciprocal heights in the two morphs, although pollen size and sculpturing do not vary. The wet, nonpapillate stigma characteristic of the species is the first to be reported in the Caesalpinioideae. In vivo and in vitro diallel crossing experiments demonstrated that a diallelic self-incompatability system exists in T. esculentum. The major site of pollen tube inhibition in the intramorph crosses was found to be in the style. This is the first report of functional heterostyly in the Fabaceae and of a confirmed self-incompatibility system in the Caesalpinioideae. Three separate lines of evidence, the monitoring of fruit development in open-pollinated plants, fruit set in diallel crossing experiments, and observations made in wild populations, demonstrated that fruit set and, by implication, seed set, are very low in this species. Floral abscission was a major limitation to the production of mature pods but there were also significant losses at other developmental stages of fruit production. The results suggest that low seed set may be an adaptation of the species to an environment in which rainfall is scarce. PMID- 12096821 TI - Expression pattern of Aux/IAA genes in the iaa3/shy2-1D mutant of Arabidopsis thaliana (L.). AB - A semi-dominant mutant suppressor of hy2 (shy2-1D) of Arabidopsis thaliana, originally isolated as a photomorphogenesis mutant, shows altered auxin responses. Recent molecular cloning revealed that the SHY2 gene is identical to the IAA3 gene, a member of the primary auxin-response genes designated the Aux/IAA gene family. Because Aux/IAA proteins are reported to interact with auxin response factors, we investigated the pattern of expression of early auxin genes in the iaa3/shy2-1D mutant. RNA hybridization analysis showed that levels of mRNA accumulation of the early genes were reduced dramatically in the iaa3/shy2-1D mutants, although auxin still enhanced gene expression in the iaa3/shy2-1D mutant. Histochemical analysis using a fusion gene of the auxin responsive domain (AuxRD) and the GUS gene showed no IAA-inducible GUS expression in the root elongation zone of the iaa3/shy2-1D mutant. On the other hand, ectopic GUS expression occurred in the hypocotyl, cotyledon, petiole and root vascular tissues in the absence of auxin. These results suggest that IAA3/SHY2 functions both negatively and positively on early auxin gene expression. PMID- 12096822 TI - Genes from the APETALA3 and PISTILLATA lineages are expressed in developing vascular bundles of the tuberous rhizome, flowering stem and flower Primordia of Eranthis hyemalis. AB - In Arabidopsis thaliana expression of the B-class MADS-box genes APETALA3 (AP3) and PISTILLATA (PI) is confined to petals and stamens but in other plant species these genes are also transcribed in non-flower tissues; in Solanum tuberosum they are transcribed specifically in vascular bundles leading to petals and stamens. Transcription analysis of B-class genes in Eranthis hyemalis using reverse transcribed in situ PCR revealed that both AP3 and PI are expressed in developing vascular bundles in the tuberous rhizome, flowering stem and floral primordia. In addition, AP3 and PI transcripts are also found in stems and leaves. These results suggest a more complex role of B-class genes in Eranthis and possible involvement in the development of vascular tissue. PMID- 12096823 TI - Secondary cell wall deposition causes radial growth of fibre cells in the maturation zone of elongating tall fescue leaf blades. AB - A gradient of development consisting of successive zones of cell division, cell elongation and cell maturation occurs along the longitudinal axis of elongating leaf blades of tall fescue (Festuca arundinacea Schreb.), a C3 grass. An increase in specific leaf weight (SLW; dry weight per unit leaf area) in the maturation region has been hypothesized to result from deposition of secondary cell walls in structural tissues. Our objective was to measure the transverse cell wall area (CWA) associated with the increase in SLW, which occurs following the cessation of leaf blade elongation at about 25 mm distal to the ligule. Digital image analysis of transverse sections at 5, 15, 45, 75 and 105 mm distal to the ligule was used to determine cell number, cell area and protoplast area of structural tissues, namely fibre bundles, mestome sheaths and xylem vessel elements, along the developmental gradient. Cell diameter, protoplast diameter and area, and cell wall thickness and area of fibre bundle cells were calculated from these data. CWA of structural tissues increased in sections up to 75 mm distal to the ligule, confirming the role of cell wall deposition in the increase in SLW (r2 = 0.924; P < or = 0.01). However, protoplast diameter of fibre cells did not decrease significantly as CWA increased, although mean thickness of fibre cell walls increased by 95 % between 15 and 105 mm distal to the ligule. Therefore, secondary cell wall deposition in fibre bundles of tall fescue leaf blades resulted in continued radial expansion of fibre cells rather than in a decrease in protoplast diameter. PMID- 12096824 TI - The pollination ecology of Grevillea beadleana McGillivray, an endangered shrub from northern New South Wales, Australia. AB - Grevillea beadleana (Proteaceae) is an endangered species known from five populations in northern New South Wales, Australia. The reproductive ecology of G. beadleana was compared in two populations with a ten-fold difference in the number of plants. Grevillea beadleana was found to be self-compatible in both populations and an examination of pollen viability and stigma maturation revealed that the species is protandrous. Flowering within inflorescences is acropetallous. In the first season plants in the largest population produced approx. ten-fold more inflorescences than those in the smaller population and, although the number of flowers per inflorescence did not vary significantly between populations the first season, the larger population produced more fruit per inflorescence than the smaller population. However, fruit to flower ratios were less than 0.2 in both seasons and populations. In both populations the number of fruit was significantly greater at the proximal end of the inflorescence, where flowers open first, compared with medial and distal positions. Several bird species were observed visiting flowers, although few birds were recorded foraging at plants in the smaller population. Within both populations, birds tended to make more within- than between-plant visits. Self compatibility, acropetally and proximal fruit-set, combined with the predominantly within-plant movement of honeyeaters, suggests inbreeding may be common within both populations of G. beadleana. Pollination and fruiting success are discussed for G. beadleana and breeding systems among rare and common taxa in Grevillea are reviewed. PMID- 12096825 TI - The effect of 2 combined oral Contraceptives containing either drospirenone or cyproterone acetate on acne and seborrhea. AB - A new oral contraceptive has been developed that contains a unique progestogen, drospirenone (DRSP), and that has both antiandrogenic and antimineralocorticoid activity. Our objective was to compare the effect of 30 microg ethinyl estradiol (EE)/3 mg DRSP (EE/DRSP; Yasmin, Schering AG, Berlin, Germany) with that of 35 microg EE/2 mg cyproterone acetate (EE/CPA; Diane-35, Schering AG, Berlin, Germany) on mild-to-moderate cases of acne. Diane-35 is used worldwide (it is not on the market in the United States and Japan) as a hormone treatment for acne, with additional contraceptive benefits. This multicenter, double-blind, randomized study was completed over 9 treatment cycles. A total of 128 women with mild-to-moderate facial acne, with or without seborrhea and/or hirsutism, were randomized. Treatment with either EE/DRSP or EE/CPA was assigned in a 2:1 ratio. Acne lesions, sebum production, and hair growth on the upper lip, chin, and chest were assessed, as well as levels of total and free testosterone, androstenedione, dehydroepiandrosterone sulfate (DHEAS), sex hormone-binding globulin (SHBG), and luteinizing hormone (LH). At study completion, dermatologists, gynecologists, and subjects gave their overall assessment of the effect of treatment on acne. After 9 treatment cycles, the median total acne lesion count was reduced markedly by 62.5% in the EE/DRSP group and 58.8% in the EE/CPA group. A comparison of the 2 groups revealed that EE/DRSP was at least as effective as EE/CPA. Both preparations also reduced sebum production and hair growth on the upper lip and chin. A 3-fold increase in the levels of SHBG was observed in both treatment groups, and levels of androgens and LH decreased. Treatment differences were not seen. Subjective evaluation of the effect of treatment on facial acne by dermatologists, gynecologists, and the subjects themselves indicated an excellent or good improvement for most subjects in both groups. EE/DRSP has been shown to be as effective for treating mild-to-moderate acne as a preparation containing EE/CPA. This new preparation may provide useful hormone therapy for women with androgen-dependent disorders who also require contraception. PMID- 12096826 TI - Risedronate preserves trabecular architecture and increases bone strength in vertebra of ovariectomized minipigs as measured by three-dimensional microcomputed tomography. AB - Risedronate reduces the risk of new vertebral fractures up to 70% within 1 year of treatment in patients with osteoporosis. Both increases in bone mass and preservation of bone architecture are thought to contribute to antifracture effects. Our objectives were to determine the effects of risedronate on trabecular bone mass and architecture and to determine the relative contributions of mass and architecture to strength in the vertebra of ovariectomized (OVX) minipigs. The minipigs were OVX at 18 months of age and were treated daily for 18 months with either vehicle or risedronate at doses of 0.5 mg/kg per day or 2.5 mg/kg per day. The three-dimensional (3D) bone architecture of the L4 vertebral cores of Sinclair S1 minipigs was evaluated by 3D microcomputed tomography (muCT). Compared with the OVX control, the vertebral bone volume (bone volume/tissue volume [BV/TV]) was higher in both treated groups (p < 0.05). The architectural changes were more significant at the 2.5-mg/kg dose and were more prevalent at the cranial-caudal ends compared with the midsection. At the higher dose, the trabecular thickness (Tb.Th), trabecular number (Tb.N), and connectivity were higher, and marrow star volume (Ma.St.V) and trabecular separation (Tb.Sp) were lower (p < 0.05). The trabecular separation variation index (TSVI), a new measure to approximate structural variations, was smaller in the 2.5-mg/kg-treated group (p < 0.05). In this group, a significant preservation of trabeculae orthogonal to the cranial-caudal axis was confirmed by a decrease in the degree of anisotropy (DA) and an increase in the percent Cross-strut (% Cross-strut; p < 0.05). Both normalized maximum load (strength) and normalized stiffness of the same vertebral cores were higher in the 2.5-mg/kg risedronate group compared with the OVX group (p < 0.05). BV/TV alone could explain 76% of the variability of the bone strength. The combination of bone volume and architectural variables explained >90% of the strength. The study showed that risedronate preserved trabecular architecture in the vertebra of OVX minipigs, and that bone strength is tightly coupled to bone mass and architecture. PMID- 12096827 TI - Bone quality: getting closer to a definition. PMID- 12096828 TI - Commentary on clinical safety of recombinant human parathyroid hormone 1-34 in the treatment of osteoporosis in men and postmenopausal women. PMID- 12096829 TI - Is lateral spine densitometric assessment equivalent to lateral radiography? PMID- 12096830 TI - Overexpression of parathyroid hormone-related protein causes hypercalcemia but not bone metastases in a murine model of mammary tumorigenesis. AB - Several lines of evidence suggest that production of parathyroid hormone-related protein (PTHrP) by breast cancer cells contributes to the formation of bone metastases. However, it is not clear if PTHrP promotes access of cancer cells to the skeleton or if it simply promotes bone resorption around cells already within bone. To study the effects of PTHrP on the development of bone metastases, we treated mice overexpressing PTHrP in their mammary glands (K14-PTHrP transgenic mice) with 9,10-dimethyl-1,2-benz-anthracene (DMBA), a known mammary carcinogen. After DMBA treatment, K14-PTHrP mice showed a higher incidence of tumor formation and a shorter latency to tumor formation than wild-type littermates. Transgenic tumors expressed the K14-PTHrP transgene and secreted excess amounts of PTHrP. In response, tumor-bearing transgenic mice became hypercalcemic and had elevated circulating levels of PTHrP. Despite the development of visceral metastases, neither transgenic mice nor wild-type controls developed bone metastases. This was true even if tumor cells were introduced into the arterial circulation of immunodeficient mice. Our results are consistent with the emerging notion that the ability of breast cancer cells to produce PTHrP in response to cues from the bone microenvironment may be more important to the development of skeletal metastases than the production of PTHrP by cells within the primary breast cancer. PMID- 12096831 TI - Bone origin of the serum complex of calcium, phosphate, fetuin, and matrix Gla protein: biochemical evidence for the cancellous bone-remodeling compartment. AB - We previously described the discovery of a fetuin-matrix Gla protein (MGP) mineral complex in the serum of rats treated with the bone-active bisphosphonate etidronate and showed that the appearance of this complex in serum correlates with the inhibition of bone mineralization by etidronate. In this study we show that the inhibition of bone resorption by treatment with the hormone calcitonin, the cytokine osteoprotegerin, or the drug alendronate, completely inhibits the generation of the fetuin-mineral complex in response to etidronate injection. These observations can be explained best by the bone-remodeling compartment (BRC), a cancellous bone compartment in which the concentrations of calcium and phosphate are determined directly by the combined actions of the osteoclast and the osteoblast. When bone mineralization is acutely inhibited by etidronate, the BRC model predicts that the continuing action of osteoclasts will cause a sharp rise in the concentrations of calcium and phosphate in the aqueous solution of the BRC with the consequent spontaneous formation of calcium phosphate crystal nuclei in which growth then would be arrested by formation of a complex with fetuin. When the inhibition of bone resorption by calcitonin, osteoprotegerin, or alendronate is combined with the acute inhibition of bone mineralization with etidronate, the BRC model correctly predicts that there will no longer be a sharp rise in calcium and phosphate, and, therefore, there will no longer be the formation of the fetuin-mineral complex. The vascular nature of the BRC is supported by the observations that the fetuin component of the fetuin-mineral complex is derived from plasma fetuin and that the fetuin mineral complex appears in plasma within minutes of the inhibition of bone mineralization with etidronate. PMID- 12096832 TI - Smad3 promotes alkaline phosphatase activity and mineralization of osteoblastic MC3T3-E1 cells. AB - Transforming growth factor (TGF) beta is abundantly stored in bone matrix and appears to regulate bone metabolism. Although the Smad family proteins are critical components of the TGF-beta signaling pathways, the roles of Smad3 in the expression of osteoblastic phenotypes remain poorly understood. Therefore, this study was performed to clarify the roles of Smad3 in the regulation of proliferation, expression of bone matrix proteins, and mineralization in osteoblasts by using mouse osteoblastic cell line MC3T3-E1 cells stably transfected with Smad3. Smad3 significantly inhibited [3H]thymidine incorporation and fluorescent intensity of the MTT-dye assay, compared with empty vector. Moreover, Smad3 increased the levels of type I procollagen, osteopontin (OPN), and matrix Gla protein (MGP) mRNA in Northern blotting. These effects of Smad3 mimicked the effects of TGF-beta on the same cells. On the other hand, Smad3 greatly enhanced ALP activity and mineralization of MC3T3-E1 cells compared with empty vector, although TGF-beta inhibited ALP activity and mineralization of wild type MC3T3-E1 cells. A type I collagen synthesis inhibitor L-azetidine-2 carboxylic acid, as well as osteocalcin (OCN), significantly antagonized Smad3 stimulated ALP activity and mineralization of MC3T3-E1 cells. In conclusion, this study showed that in mouse osteoblastic cells, Smad3 inhibited proliferation, but it also enhanced ALP activity, mineralization, and the levels of bone matrix proteins such as type I collagen (COLI), OPN, and MGP. We propose that Smad3 plays an important role in osteoblastic bone formation and might help to elucidate the transcriptional mechanism of bone formation and possibly lead to the development of bone-forming drugs. PMID- 12096833 TI - NF-kappaB p50 and p52 expression is not required for RANK-expressing osteoclast progenitor formation but is essential for RANK- and cytokine-mediated osteoclastogenesis. AB - Expression of RANKL by stromal cells and of RANK and both NF-kappaB p50 and p52 by osteoclast precursors is essential for osteoclast formation. To examine further the role of RANKL, RANK, and NF-KB signaling in this process, we used NF kappaB p50-/- ;p52-/- double knockout (dKO) and wild-type (WT) mice. Osteoclasts formed in cocultures of WT osteoblasts with splenocytes from WT mice but not from dKO mice, a finding unchanged by addition of RANKL and macrophage colony stimulating factor (M-CSF). NF-kappaB dKO splenocytes formed more colony-forming unit granulocyte macrophage (CFU-GM) colonies than WT cells, but no osteoclasts were formed from dKO CFU-GM colonies. RANKL increased the number of CFU-GM colonies twofold in WT cultures but not in dKO cultures. Fluorescence-activated cell sorting (FACS) analysis of splenocytes from NF-kappaB dKO mice revealed a two-to threefold increase in the percentage of CD11b (Mac-1) and RANK double positive cells compared with WT controls. Treatment of NF-kappaB dKO splenocytes with interleukin (IL)-1, TNF-alpha, M-CSF, GM-CSF, and IL-6 plus soluble IL-6 receptor did not rescue the osteoclast defect. No increase in apoptosis was observed in cells of the osteoclast lineage in NF-kappaB dKO or p50-/-;p52+/- (3/4KO) mice. Thus, NF-kappaB p50 and p52 expression is not required for formation of RANK-expressing osteoclast progenitors but is essential for RANK expressing osteoclast precursors to differentiate into TRAP+ osteoclasts in response to RANKL and other osteoclastogenic cytokines. PMID- 12096834 TI - Dual modulation of osteoclast differentiation by lipopolysaccharide. AB - Lipopolysaccharide (LPS) modulates bone resorption by augmentation of osteoclastogenesis. It increases in osteoblasts the production of RANKL, interleukin (IL)-1, prostaglandin E2 (PGE2), and TNF-alpha, each known to induce osteoclast activity, viability, and differentiation. We examined the role of direct interactions of LPS with osteoclast precursors in promoting their differentiation. To this end, we have used bone marrow mononuclear cell preparations in the absence of osteoblasts or stromal cells. We found that LPS does not induce osteoclast differentiation in these cells. Moreover, the inclusion of LPS blocked the osteoclastogenic activity of RANKL. However, LPS is a potent inducer of osteoclastogenesis in RANKL-pretreated cells, even if present in the absence of exogenous RANKL. Osteoprotegerin (OPG), does not affect the stimulatory phase of LPS modulation of osteoclastogenesis, ruling out involvement of endogenous RANKL. LPS induces the expression of TNF-a and IL-1beta in osteoclast precursors, regardless if they were or were not pretreated with RANKL. These two cytokines induced osteoclast differentiation in RANKL-pretreated cells. To examine if these cytokines mediate LPS effect in an autocrine mechanism, we measured the effect of their neutralization on LPS osteoclastogenic activity. Although neutralization of IL-1beta did not affect LPS activity, a marked inhibition was observed when TNF-alpha was neutralized. However, TNF-a expression was increased also in conditions in which LPS inhibited RANKL osteoclastogenic activity. We found that LPS reduces the expression of RANK and macrophage colony stimulating factor (M-CSF) receptor. In summary, LPS impacts on osteoclastogenesis also via its interactions with the precursor cells. LPS inhibits RANKL activity by reducing the expression of RANK and M-CSF receptor and stimulates osteoclastogenesis in RANKL-pretreated cells via TNF-alpha. PMID- 12096835 TI - Melatonin at pharmacologic doses increases bone mass by suppressing resorption through down-regulation of the RANKL-mediated osteoclast formation and activation. AB - This study evaluated if melatonin would increase bone mass in mice. Four groups of 4-week-old male ddy mice received daily injections of vehicle or 1, 5, or 50 mg/kg of melatonin, respectively, for 4 weeks. Treatment with 5 mg/kg per day or 50 mg/kg per day of melatonin significantly increased bone mineral density (BMD; by 36%, p < 0.005) and bone mass (bone volume per tissue volume [BV/TV] by 49%, p < 0.01, and trabecular thickness [Tb.Th] by 19%, p < 0.05). This treatment significantly reduced bone resorption parameters (i.e., osteoclast surface [Oc.S/bone surface [BS]] by 74%,p < 0.05, and osteoclast number [N.Oc/BS] by 76%,p < 0.005) but did not increase histomorphometric bone formation parameters (i.e., bone formation rate [BFR/ BS], mineral apposition rate [MAR], and osteoid volume [OV/TV]), indicating that melatonin increases bone mass predominantly through suppression of bone resorption. Melatonin (1-500 microM) in vitro caused dose-dependent reduction (p < 0.001 for each) in the number and area of resorption pits formed by osteoclasts derived from bone marrow cells but not those formed by isolated rabbit osteoclasts. Because RANKL increases, while osteoprotegerin (OPG) serves as a soluble decoy receptor for RANKL to inhibit osteoclast formation and activity, the effect of melatonin on the expression of RANKL and OPG in mouse MC3T3-E1 osteoblastic cells was investigated. Melatonin (5 500 microM) increased in a dose-dependent manner and reduced the mRNA level of RANKL and both mRNA and protein levels of OPG in MC3T3-E1 cells (p < 0.001 for each). In summary, these findings indicated for the first time that melatonin at pharmacologic doses in mice causes an inhibition of bone resorption and an increase in bone mass. These skeletal effects probably were caused by the melatonin-mediated down-regulation of the RANKL-mediated osteoclast formation and activation. PMID- 12096836 TI - Onion and a mixture of vegetables, salads, and herbs affect bone resorption in the rat by a mechanism independent of their base excess. AB - Prevention of low bone mass is important to reduce the incidence of osteoporotic fractures. In man, the consumption of fruits and vegetables is associated with greater bone mineral density (BMD), an effect that is claimed to be caused by their base excess buffering metabolic acid, thought to dissolve bone. We showed previously that in the rat the consumption of several vegetables, salads, and herbs inhibits bone resorption and that onion increases bone mass. In this study we show that, although the intake of onion is associated with a decrease in urinary noncarbonic acid excretion and a concomitant inhibition of bone resorption of similar magnitude, the two findings are not causally related. Onion retains its bone resorption inhibitory activity in the rat even when added to a vegetarian diet with typical base excess. Onion and a mixture of vegetables, salads, and herbs retain their inhibitory activity even when metabolic acid is buffered with potassium citrate. In addition, neither the pH nor the potassium content of individual ashed vegetables, salads, and herbs correlates with inhibition of bone resorption. The effect of vegetables, salads, and herbs, which inhibit bone resorption in the rat, therefore is not mediated by their base excess but possibly by a pharmacologically active compound(s). PMID- 12096837 TI - International variations in hip fracture probabilities: implications for risk assessment. AB - It is recommended that intervention thresholds should be based on absolute fracture risk, but there is a large variation in hip fracture incidence from different regions of the world. The aim of this study was to examine heterogeneity of hip fracture probability in different regions from recent estimates of hip fracture incidence and mortality to adjust intervention thresholds. Ten-year probabilities of hip fracture were computed in men and women at 10-year intervals from the age of 50 years and lifetime risks at the age of 50 years from the hazard functions of hip fracture and death. Lifetime risk at the age of 50 years varied from 1% in women from Turkey to 28.5% in women from Sweden. High lifetime risks in women were associated with high lifetime risks in men (r = 0.83). There also were significant correlations of 10-year risk at any age between men and women. Ten-year probability was standardized to that of men and women from Sweden (set at 1.0). There was a 15-fold range in 10-year probability from 1.24 in Norway to 0.08 in Chile. Countries were categorized by 10-year probabilities comprising very high risk (Norway, Iceland, Sweden, Denmark, and the United States), high risk (China [Taiwan [TW]], Germany, Switzerland, Finland, Greece, Canada, The Netherlands, Hungary, Singapore, Italy, United Kingdom, Kuwait, Australia, and Portugal), medium risk (China [Hong Kong [HK]], France, Japan, Spain, Argentina, and China), and low risk (Turkey, Korea, Venezuela, and Chile). The categorization of hip fracture probabilities can be used to adjust intervention thresholds based on age, sex, and relative risk from a reference population such as Sweden. PMID- 12096838 TI - Polymorphisms in the osteoprotegerin gene are associated with osteoporotic fractures. AB - Osteoprotegerin (OPG) is a soluble receptor for RANKL and therefore a competitive inhibitor of osteoclast differentiation and activity. With this key role in the control of resorptive activity, we found that OPG is a candidate gene for genetic control of bone mass. We examined the promoter and the five exons with surrounding intron sequences of the OPG gene for polymorphisms in 50 normal patients and 50 patients with osteoporosis. We found 12 polymorphisms. Two sets of four and five polymorphisms, respectively, were in complete linkage. Subsequently, we examined the effect of the informative polymorphisms A163-G (promoter), T245-G (promoter), T950-C (promoter), G1181-C (exon 1), and A6890-C (intron 4) on the prevalence of osteoporotic fractures, bone mass, and bone turnover in 268 osteoporotic patients and 327 normal controls. In A163-G the variant allele G was more common among fracture patients: 34.0% versus 26.3% in normal controls (p < 0.05) and the odds ratio (OR) for a vertebral fracture, if an individual has the G allele, was 1.44 (1.00-2.08). In T245-G the variant allele G was more common in osteoporotic patients: 12.4% versus 6.5% (p < 0.02) and the OR for vertebral fracture, if an individual has the G-allele, was 2.00 (1.10-3.62). G1181-C is located in the first exon and causes a shift in the third amino acid from lysine to asparagine. The CC genotype was less common among fracture patients: 26.3% versus 36.7% in the normal controls (p < 0.01). T950-C and A6890-C were not distributed differently among patients with osteoporosis and normal controls. None of the polymorphisms affected bone mineral density (BMD) or biochemical markers of bone turnover in the normal controls. In conclusion, we have examined the human OPG gene for polymorphisms and found 12. The rare alleles of the A163-G and T245-G were significantly more common among patients with vertebral fractures. PMID- 12096840 TI - Dietary essential amino acid supplements increase bone strength by influencing bone mass and bone microarchitecture in ovariectomized adult rats fed an isocaloric low-protein diet. AB - This study was designed to investigate whether the administration of dietary essential amino acid supplements in adult rats made osteoporotic by estrogen deficiency and reduced protein intake could reverse the deleterious effects caused by these maneuvers. This animal model was selected to mimic the situation observed in elderly women in whom estrogen deficiency and/or low-protein intake (but also calcium and vitamin D deficiency) are known to contribute to the pathogenesis of osteoporosis. Six-month-old rats were ovariectomized (OVX) and fed an isocaloric 2.5% casein diet for 10 weeks or sham-operated (SHAM) and fed an isocaloric 15% casein diet. The animals fed the 2.5% casein diet were given isocaloric supplements of essential amino acids in similar relative proportion to that of casein at doses of 2.5% or 5% of total diet for an additional 16 weeks. Vertebrae, femur, and tibia bone mineral density (BMD); ultimate strength; and microtomographic histomorphometry were evaluated before and after dietary essential amino acid supplements. Essential amino acid supplements increased vertebrae, femur, and tibia bone strength in OVX rats fed a low-protein diet. The mechanical changes induced by this dietary isocaloric supplement were associated with the prevention of a further BMD decrease or even with some increases and changes in microarchitecture such as from a rod to a plate trabecular spacial configuration and increased cortical thickness. Higher insulin-like growth factor (IGF) I levels, as well as greater bone formation and reduced bone resorption as assessed by biochemical markers of bone remodeling, were found in rats receiving essential amino acid supplements. In conclusion, dietary essential amino acid supplements increased bone strength through modifications of BMD, trabecular architecture, and cortical thickness possibly by an IGF-I-mediated process. PMID- 12096839 TI - Osteoprotegerin abrogates chronic alcohol ingestion-induced bone loss in mice. AB - To investigate the role of osteoprotegerin (OPG) on alcohol (ethanol)-mediated osteoporosis, we measured a variety of bone remodeling parameters in mice that were either on a control diet, an ethanol (5%) diet, or an ethanol (5%) diet plus OPG administration. OPG diminished the ethanol-induced (1) decrease in bone mineral density (BMD) as determined by dual-energy densitometry, (2) decrease in cancellous bone volume and trabecular width and the increase of osteoclast surface as determined by histomorphometry of the femur, (3) increase in urinary deoxypyridinolines (Dpd's) as determined by ELISA, and (4) increase in colony forming unit-granulocyte macrophage (CFU-GM) formation and osteoclastogenesis as determined by ex vivo bone marrow cell cultures. Additionally, OPG diminished the ethanol-induced decrease of several osteoblastic parameters including osteoblast formation and osteoblast culture calcium retention. These findings were supported by histomorphometric indices in the distal femur. Taken together, these data show that OPG diminishes ethanol-induced bone loss. Furthermore, they suggest that OPG achieves this through its ability to abrogate ethanol-induced promotion of osteoclastogenesis and promote osteoblast proliferation. PMID- 12096841 TI - Genetic and environmental determinants of peak bone mass in young men and women. AB - Peak bone mass is an important risk factor for the development of osteoporosis in later life. Previous work has suggested that genetic, intrauterine, and environmental factors all contribute to the regulation of bone mass, but the ways in which they interact with each other to do so remain poorly understood. In this study, we investigated the relationship between peak bone mass and polymorphisms of the vitamin D receptor (VDR), estrogen receptor (ER) a, and collagen type Ialpha1 (COLIA1) genes in relation to other factors such as birth weight, lifestyle diet, and exercise in a population-based cohort of 216 women and 244 men in their early 20s. Stepwise multiple regression analysis showed that body weight was the strongest predictor of bone mineral density (BMD) in women, accounting for 16.4% of the variance in spine BMD and 8.4% of the variance in femoral neck BMD. Other significant predictors were VDR genotype (3.8%) and carbohydrate intake (1.6%) at the spine and vitamin D intake (3.4%) and ER genotype (3.4%) at the femoral neck. Physical activity was the strongest predictor of BMD in men, accounting for 6.7% of the variance at the spine and 5.1% at the hip. Other significant predictors were body weight (5%) and ER PvuII genotype (2.8%) at the spine and weight (3.4%) and alcohol intake (2%) at the femoral neck. Birth weight was not a significant predictor of BMD at either site but COLIA1 genotype significantly predicted birth weight in women, accounting for 4.3% of the variance. We conclude that peak bone mass is regulated by an overlapping but distinct set of environmental and genetic influences that differ in men and women. However, much of the variance in BMD was unexplained by the variables studied here, which suggests that either most of the genes that regulate BMD remain to be discovered or major environmental influences on BMD exist that have not yet been identified. PMID- 12096842 TI - Systemic regulation of distraction osteogenesis: a cascade of biochemical factors. AB - This study investigates the systemic biochemical regulation of fracture healing in distraction osteogenesis compared with rigid osteotomy in a prospective in vivo study in humans. To further clarify the influence of mechanical strain on the regulation of bone formation, bone growth factors (insulin-like growth factor [IGF] I, IGF binding protein [IGFBP] 3, transforming growth factor [TGF] beta1, and basic FGF [bFGF]), bone matrix degrading enzymes (matrix-metalloproteinases [MMPs] 1, 2, and 3), human growth hormone (hGH), and bone formation markers (ALP, bone-specific ALP [BAP], and osteocalcin [OC]) have been analyzed in serum samples from 10 patients in each group pre- and postoperatively. In the distraction group, a significant postoperative increase in MMP-1, bFGF, ALP, and BAP could be observed during the lengthening and the consolidation period when compared with the baseline levels. Osteotomy fracture healing without the traction stimulus failed to induce a corresponding increase in these factors. In addition, comparison of both groups revealed a significantly higher increase in TGF-beta1, IGF-I, IGFBP-3, and hGH in the lengthening group during the distraction period, indicating key regulatory functions in mechanotransduction. The time courses of changes in MMP-1, bone growth factors (TGF-beta1 and bFGF), and hGH, respectively, correlated significantly during the lengthening phase, indicating common regulatory pathways for these factors in distraction osteogenesis. Significant correlation between the osteoblastic marker BAP, TGF beta1, and bFGF suggests strain-activated osteoblastic cells as a major source of systemically increased bone growth factors during callus distraction. The systemic increase in bFGF and MMP-1 might reflect an increased local stimulation of angiogenesis during distraction osteogenesis. PMID- 12096843 TI - Association analysis of single nucleotide polymorphisms in cartilage-specific collagen genes with knee and hip osteoarthritis in the Japanese population. AB - Osteoarthritis (OA) is one of the most common diseases in the elderly. Although its pathophysiology is complex and its molecular basis remains to be determined, much evidence suggests that OA has strong genetic determinants. To search for susceptibility loci of OA, we selected seven candidate genes encoding cartilage specific collagens (type II, IX, X, and XI collagens) and performed association analysis for OA using single nucleotide polymorphisms (SNPs) in the coding region of these genes. Four hundred seventeen OA samples and 280 control samples were collected from the Japanese population, and 12 SNPs were genotyped. Our studies have identified two susceptibility loci of OA: COL2A1 and COL9A3. An SNP in COL9A3 showed significant association with knee OA (p = 0.002, odds ratio [OR] = 1.48). Haplotype analysis showed significant association between a specific haplotype of COL2A1 and hip OA (p = 0.024; OR = 1.30). Further analysis of these two genes will shed light on the molecular mechanisms of OA. PMID- 12096844 TI - Impaired vascular invasion of Cbfa1-deficient cartilage engrafted in the spleen. AB - Chondrocyte maturation and vascular invasion of cartilage are essential in the process of endochondral ossification. Cbfal-deficient (Cbfa1-/-) mice displayed a complete absence of osteoblast and osteoclast maturation as well as severely inhibited chondrocyte maturation in most parts of the skeleton. Although chondrocyte maturation and mineralization were observed in restricted areas of Cbfa1-/- mouse skeleton, vascular invasion of calcified cartilage was never noted. To investigate the possibility of chondrocyte maturation and vascular invasion in Cbfal-/- cartilage and the role of the hematopoietic system in the process of vascular invasion, we transplanted embryonic day 18.5 (E18.5) Cbfa1-/- femurs, which are composed of immature chondrocytes, into spleens of normal mice. One week later, the transplanted femurs contained terminally differentiated chondrocytes expressing osteopontin, bone sialoprotein (BSP), and matrix metalloproteinase (MMP) 13. In the diaphyses of the transplants, the cartilage matrix was mineralized and the cartilage was invaded by vascular vessels and osteoclasts. However, chondrocyte maturation and vascular invasion were severely retarded in comparison with transplants of E14.5 wild-type femurs, in which the cartilage was rapidly replaced by bone, and neither mature osteoblasts nor bone formation were observed. In primary culture of Cbfa1-/- chondrocytes, transforming growth factor (TGF) beta1, platelet-derived growth factor (PDGF), interleukin (IL)-1beta, and thyroid hormone (T3) induced osteopontin and MMP-13 expression. These findings indicated that factors in the hematopoietic system are able to support vascular invasion of cartilage independent of Cbfal but are less effective without it, suggesting that Cbfal functions in cooperation with factors from bone marrow in the process of growth plate vascularization. PMID- 12096845 TI - Size of bone in the aged endocortical resorption. PMID- 12096847 TI - Satellite remote sensing as a tool in Lahar disaster management. AB - At least 40,000 deaths have been attributed to historic lahars (volcanic mudflows). The most recent lahar disaster occurred in 1998 at Casita volcano, Nicaragua, claiming over 2,500 lives. Lahars can cover large areas and be highly destructive, and constitute a challenge for disaster management. With infrastructure affected and access frequently impeded, disaster management can benefit from the synoptic coverage provided by satellite imagery. This potential has been recognisedfor other types of natural disasters, but limitations are also known. Dedicated satellite constellations for disaster response and management have been proposed as one solution. Here we investigate the utility of currently available and forthcoming optical and radar sensors as tools in lahar disaster management. Applied to the Casita case, we find that imagery available at the time could not have significantly improved disaster response. However, forthcoming satellites, especially radar, will improve the situation, reducing the benefit of dedicated constellations. PMID- 12096846 TI - Civil society and the state: Turkey after the earthquake. AB - On 17 August 1999 Turkey was hit by a massive earthquake. Over 17,000 lives were lost and there was extensive damage to Turkey's heartland. This paper examines how various public and private institutions, including state and civil society institutions such as NGOs and the media responded to the needs of earthquake survivors. It documents the extensive involvement of NGOs in the relief efforts immediately after the disaster and examines the impact of such participation on state-civil society relations in the country. The data show that state response to the disaster went through several phases from a period of ineptitude to effective management. The paper credits the media and the NGOs for acting as advocates for survivors and forcing changes at the state level. The paper argues that an ideal response system, which fully addresses the needs of victims, can only be based on state-civil society relations that are both collaborative and adversarial. PMID- 12096848 TI - Risk removed? Steps towards building trust in humanitarian mine action. AB - This article examines the role of building trust in Humanitarian Mine Action (HMA). The role of trust following a mine-clearance operation is largely an unexplored region. Drawing on case studies from Angola, Mozambique and Afghanistan, the article explores how trust can be understood in the context of humanitarian initiatives such as landmine clearance, and outlines the challenges associated with building trust among populations with experience of armed conflict. Because trust is difficult to re-establish following a war, the argument presented here suggests that one way in which this can be done is through building relationships at the organisational and interpersonal levels. The analysis reviews specific measures that mine-action agencies have used to build trust, looking at the strengths and weaknesses of different approaches. The analysis concludes by suggesting general guidelines for how mine-action practitioners can facilitate trust, and by outlining some promising avenues for future research. PMID- 12096849 TI - Measuring the impact of rehabilitation services on the quality of life of disabled people in Cambodia. AB - The present study aimed to measure the quality of life (QOL) of disabled people in Cambodia and the impact of rehabilitation services. The categories of services were: physical rehabilitation (prosthetics/orthotics with physiotherapy); community-based rehabilitation; and (3) labour market assistance. The 164 respondents were from a range of urban and rural settings. The results suggest that QOL was substantially lower among disabled Cambodians who had received no rehabilitation services compared to those who had received one of the three categories of rehabilitation services. QOL scores tended to be highest, however, among those who had received a combination of all three services. The results also highlighted the vulnerability of certain sub-groups who may need specific provision in planning, policy-making and service delivery. The present study suggests that an integrated approach may be the best way to maximise the impact of individual rehabilitation services in Cambodia. PMID- 12096850 TI - Guatemala: violence in peacetime--a critical analysis of the armed conflict and the peace process. AB - The recent history of Guatemala is interpreted here using an interdisciplinary approach inspired by recent development and conflict studies. The author reflects on his experience in peace-building activities and uses a combination of primary and secondary sources to present the Guatemalan war as a complex political emergency. The first part (a methodological introduction) proposes an analytical framework, taking into account both the root causes and the different functions of violence, in order to achieve a better understanding of contemporary conflicts. In the second and central part of the article, the integrated framework is applied to Guatemala, trying to understand causes and functions of war and negotiation and underlining the existence of structural obstacles to a lasting peace. The conclusions are dedicated to policy implications: if the signing of peace accords represents only a step towards negative peace, the condition for a lasting peace is a strategy for reconciliation and development based on social participation and social justice. PMID- 12096851 TI - Imaging in scoliosis: what, why and how? AB - Scoliosis may be a spinal manifestation of underlying disease and although most cases of scoliosis are idiopathic, imaging plays a very important role in determining the underlying aetiology and in monitoring the changes of the deformity that take place with growth. As a clinical problem scoliosis may present directly to the radiology department through a primary healthcare referral, or it may be referred from the paediatric, orthopaedic, spinal and neurosurgical hospital services. Growth affects all types of scoliosis irrespective of cause. There are no reliable predetermined algorithmic steps in the management of scoliosis, and treatment decisions require the inclusion of multiple extrinsic (e.g age, menarche) and intrinsic (curve magnitude, vertebral anomaly) factors. It is important to remember that most of what is known about curve behaviour and its progression applies to idiopathic scoliosis, and it is inappropriate to apply these criteria to the other specific types of scoliosis. It is imperative that radiation techniques are used judiciously to minimize the radiation burden. Although magnetic resonance imaging (MRI) has had a significant impact in the understanding of the scoliotic deformity, it is still evolving and it may well eventually play a very important role in uncovering the underlying aetiology of 'idiopathic' scoliosis. PMID- 12096852 TI - Hormone replacement therapy and mammographic screening. AB - The benefits of hormone replacement therapy (HRT) with oestrogen are well known and have led to widespread usage of HRT in post-menopausal women. There is an increased relative risk of breast cancer with prolonged HRT use of 1.7 at worst and this does not warrant more frequent screening. HRT itself makes mammographic screening less effective by adversely affecting both the sensitivity and specificity of screening mammography.A number of large studies have shown a reduction in the sensitivity of screening mammography of between 7% and 21% in current HRT users. This reduction in sensitivity is seen only in women over the age of 50 and is more marked when using single view mammography. The reduction in sensitivity is largely confined to those women who have a dense mammographic background pattern while on HRT. HRT use is also associated with a reduction in specificity of between 12% and almost 50%. This reduction in specificity is mainly found at incident screens. What can be done to minimize the adverse effects of HRT on mammographic screening? The imminent introduction of two views at all screens within the National Health Breast Screening Programme will be helpful. Short-term cessation of HRT use may deter attendance for screening by HRT users and is unproven. Combined oestrogen and progesterone preparations taken continuously appear to be particularly associated with adverse breast screening performance, while tibolone may have little effect on mammographic density. Manipulation of the type of HRT preparations used may allow the adverse effects of HRT on breast screening to be reduced. PMID- 12096853 TI - Nuclear medicine in primary breast cancer imaging. AB - The application of nuclear medicine techniques to study patients with breast cancer has recently raised its profile, particularly in the investigation 'indeterminate mammographic lesions'. This review briefly points out some of the difficulties encountered with other more conventional imaging modalities and describes the radionuclide techniques most frequently employed in the investigation of those patients with breast cancer. Both planar and single photon emission tomography methods are discussed including the use of monoclonal antibodies, perfusion ligands, receptor binding hormones and other specific radiotracers, non-specific tumour markers, as well as deoxyglucose and other amino acids labelled with positron emitting radionuclides. PMID- 12096854 TI - Effects of mailed dissemination of the Royal College of Radiologists' guidelines on general practitioner referrals for radiography: a time series analysis. AB - AIM: To evaluate the effect of postal dissemination of the third edition of the Royal College of Radiologists' (RCR) guidelines on general practitioner referrals for radiography. MATERIALS AND METHODS: An interrupted time series using monthly data for 34 months before and 14 months after dissemination of the guidelines was employed. Data were abstracted for the period April 1994 to March 1998 from the computerized administrative systems of open access radiological services provided by two teaching hospitals in one region of Scotland. The time series results are contrasted with those obtained by using a simple before and after design. RESULTS: A total of 117 747 imaging requests from general practice were received in the two departments. There were no significant effects of disseminating the guidelines on the total number of requests, or on requests for individual examinations. If a simple before and after study had been used, then we would have erroneously concluded that significant changes had occurred in referral practice for 11 of the 18 procedures concerned. CONCLUSION: Mailing of copies of the RCR guidelines had a small effect on general practitioners' use of X-ray investigations of uncertain clinical significance. Additional dissemination and implementation strategies appear necessary to promote the use of guidelines. PMID- 12096855 TI - Electrical impedance scanning of lymph nodes: initial clinical and technical findings. AB - PURPOSE: The sonographic differentiation between inflammatory and malignant lymph node enlargement remains imprecise. Electrical impedance scanning (EIS) is of more value in malignant breast lesions, and this study was undertaken to determine if it was of use in differentiating nodal enlargement. MATERIALS AND METHODS: Two hundred and thirty-six sonographically suspicious lymph nodes (146 patients) of mean size 18 x 11 x 11 mm mean depth 9 mm were examined using a commercially available electrical impedance imaging system. The following locations of lymph nodes were included in the study: cervical, axillary, inguinal, supraclavicular and chest/abdominal wall. The EIS results were compared to histopathological, serological or follow up findings. RESULTS: Using EIS 105/115 malignant nodes were correctly diagnosed, while 70/121 inflammatory or benign lymph nodes were correctly identified as benign by EIS (10 false-negative, 51 false-positive). Sensitivity (91.3%) and specificity (57.9%) were obtained. Corresponding negative and positive predictive values were 87.5% and 67.3%, respectively. CONCLUSIONS: Results from this initial study suggest potential value for EIS as an imaging adjunct in the differentiation of sonographically equivocal lesions. Best accuracy was achieved at chest/abdominal wall and inguinal locations. Due to technical restrictions with the present system, the examination of inframandibular and para-aortal lymph nodes should be limited to special cases. PMID- 12096856 TI - Use of stents in the palliative treatment of malignant gastric outlet and duodenal obstruction. AB - AIM: To evaluate the efficacy of stenting in the palliation of malignant duodenal and gastric outlet obstruction. MATERIALS AND METHODS: We retrospectively reviewed our series of patients who underwent stenting for malignant upper gastrointestinal obstruction between March 1998 and December 1999. From January 2000 data have been acquired prospectively. Our series comprises 21 stents successfully deployed in 15 patients. RESULTS: The technical and clinical success was 93% (14/15 patients). One patient required endoscopic negotiation of recurrent gastric carcinoma at the gastrojejunostomy site after failure to cross the lesion fluroscopically. Two patients required re-intervention 2 and 5 weeks after initial stent placement, for migration and ingrowth respectively. Eighteen stents were placed transorally, two stents transhepatically and one via a transgastric approach. Early complications (pain < 3 days) occurred in two patients (13%) and late complications (ingrowth, overgrowth and migration) occurred in three patients (20%). The median survival was 2.4 months (range 2-4 months). CONCLUSION: Stenting provides a less invasive palliative option than surgery with the advantage of lower morbidity and complication rates. It has the advantage of high technical and clinical success rates facilitated by alternative routes of access into the upper gastrointestinal tract via transgastric and transhepatic routes in addition to the traditional peroral route. PMID- 12096857 TI - Restoration of myocardial blood flow following percutaneous coronary balloon dilatation and stent implantation: assessment with qualitative and quantitative contrast-enhanced magnetic resonance imaging. AB - AIM: To examine the serial use of magnetic resonance imaging (MRI) to evaluate regional myocardial perfusion changes following percutaneous coronary angioplasty and stent implantation (PTCA). MATERIALS AND METHODS: Six patients with single vessel coronary artery disease (CAD) underwent contrast-enhanced first pass MRI immediately prior to (visit A) and within 7 days after (visit B) PTCA. Three sequential short axis slices were obtained after gadodiamide (Gd) bolus (0.025 mmol/kg(-1)) at rest and during adenosine. Each short axis was divided radially into eight regions of interest (ROIs). ROIs were anatomically assigned to a coronary artery territory (CAT). Stress and rest qualitative and quantitative (unidirectional extraction fraction constant (K(i)); index of myocardial perfusion reserve (MPRI) = stressK(i) / restK(i)) perfusion parameters were determined for ROI supplied by remote and stenosed/stented vessels for each visit. RESULTS: In stented ROIs the number of ROIs demonstrating normal perfusion, as opposed to reversible perfusion deficits, increased. Qualitative perfusion assessment in remote CATs was unchanged. MPRI in stenotic CATs was lower than in remote CATs at visit A (P < 0.001). Following PTCA, MPRI increased in stented CATs (P < 0.001) but was unchanged in remote CATs. CONCLUSION: Restoration of myocardial perfusion following PTCA can be delineated with qualitative and quantitative perfusion MRI. Although at present the investigation is technically complex and not perfectly sensitive or specific, MRI has the potential to be a valuable tool for patient follow-up and evaluation of revascularization strategy efficacy. PMID- 12096858 TI - Carotid intima-media thickness in patients with head and neck irradiation for the treatment of nasopharyngeal carcinoma. AB - AIM: Intima-media thickness (IMT) has been shown to be useful in the evaluation and monitoring of carotid artery atherosclerosis in patients at risk of cardiovascular events. In this study, we aimed to examine the IMT in patients with nasopharyngeal carcinoma (NPC) who received irradiation to the carotid arteries during radiotherapy, and compared them with a control group. MATERIALS AND METHODS: Fifty-one NPC patients (aged between 39 and 69 years) and a group of 51 age-and sex-matched controls were studied by ultrasound. The IMT at the far wall of the common carotid artery was measured three times and the average value taken in each subject. The IMT of both groups were compared. Risk factors for IMT thickness, including hypertension, smoking, hyperglycaemia, hypercholesterolaemia, history of cerebrovascular accidents and cardiovascular disease, were also studied. RESULTS: The mean carotid IMT of patients in the NPC group (2.2+/-1.5 mm) was statistically greater than that in normal controls (0.7+/-0.15 mm) (P < 0.05). There was no statistically significant difference between the IMT in the right and left common carotid arteries within each group of patients. CONCLUSION: The findings suggest that patients with irradiation have increased arterial IMT. As they are asymptomatic the clinical relevance is not clear. PMID- 12096859 TI - Double contrast barium enema sensitivity: a comparison of studies by radiographers and radiologists. AB - PURPOSE: A retrospective study of histologically proven cases of colorectal cancer (CRC) was performed to assess whether the sensitivity of the radiographer performed double contrast barium enema (DCBE) differed from that of the radiologist-performed study. MATERIALS AND METHODS: Histologically proven cases of CRC were reviewed over a 3-year period to ascertain whether: the diagnosis had been made by DCBE in the 3 years before histological diagnosis; the lesion had been correctly diagnosed; the examination had been performed by a radiologist or radiographer. RESULTS: In the 3-year period there were 478 cases with histologically proven CRC. Of these, 239 (50%) had undergone DCBE as the initial radiological investigation of the colon. Sixty-four examinations had been performed by radiographers. A correct diagnosis was made in 58 cases (90.6%), the report was equivocal in one case (1.6%), there were four false-negatives (6.25%), and one case was abandoned (1.6%). One hundred and seventy-five examinations were performed by radiologists. A correct diagnosis was made in 157 cases (89.7%), the report was equivocal in one case (0.6%), there were 16 false-negatives (9.1%), and one case was abandoned (0.6%). CONCLUSION: A sensitivity of 90.6% for radiographer-performed studies compared favourably with 89.7% for radiologist performed studies and supports the practice of radiographers undertaking barium enemas. PMID- 12096860 TI - CT in pseudomyxoma peritonei: a review of 17 cases. AB - AIM: To describe the computed tomography (CT) findings in pseudomyxoma peritonei. MATERIALS AND METHOD: Two observers independently and retrospectively reviewed the CT images of 17 consecutive patients (nine women, eight men, mean age 53 years) with histologically proven pseudomyxoma peritonei. RESULTS: Six patients had small volume disease where pseudomyxoma peritonei was present in focal collections in the peritoneal cavity. Eleven had large volume disease that completely, or almost completely, filled the peritoneal cavity. Pseudomyxoma peritonei is characterized by low attenuation mucinous ascites on CT. Areas of high attenuation, septae and calcification are seen more commonly within it as the volume of disease increases. The pattern of accumulation of pseudomyxoma peritonei follows the normal flow of peritoneal fluid. It initially seeds at sites of relative stasis and as large volume disease develops it fills the remaining spaces in the peritoneal cavity and pressure effects dominate imaging. Pseudomyxoma peritonei may extend into hernial orifices or the pleural cavity. CONCLUSION: Pseudomyxoma peritonei is difficult to diagnose clinically. However, the pattern of accumulation of disease is predictable and can be recognized on CT. PMID- 12096861 TI - Scattered radiation level during videofluoroscopy for swallowing study. AB - AIM: To evaluate the scattered radiation exposure to the surroundings during videofluoroscopy for swallowing study (VFSS). MATERIALS AND METHODS: Scattered radiation exposure was measured using an ion chamber survey meter for 17 adult patients undergoing videofluoroscopy for swallowing study. The cumulative dose area product of each case was also recorded. Data were presented as mean +/- standard deviation. RESULTS: The scattered radiation exposure at a distance of 150 cm from the patient and the dose area product recorded were 149 +/- 78 microR (range 42-308 microR) and 842 +/- 544 cGy.cm(2) (range 258-2151 cGy.cm(2)), respectively, for a single study of 18 +/- 6 minutes. A formula was then derived for estimating the scattered radiation dose to muscle tissue at an arbitrary distance based on the accumulated dose area product. With this formula, the mean scattered radiation dose to naked muscle tissue of the surrounding people at a distance of 30-100 cm from the patient were estimated to be 33.68-3.03 microSv respectively. CONCLUSION: The scattered radiation detriment associated with videofluoroscopy for swallowing study was well within acceptable levels. PMID- 12096862 TI - Magnetic resonance angiography for the diagnosis of renal artery stenosis: a meta analysis. AB - AIM: To review the published literature comparing the diagnostic accuracy of magnetic resonance angiography (MRA) with and without gadolinium in diagnosing renal artery stenosis, using catheter angiography as reference. MATERIALS AND METHODS: A meta-analysis was performed of English language articles identified by computer search using PubMed/MEDLINE, followed by extensive bibliography review from 1985 to May 2001. Inclusion criteria were: (1) blinded comparison with catheter angiography; (2)indication for MRA stated; (3) clear descriptions of imaging techniques; and (4) interval between MRA and catheter angiography < 3 months and only the largest of all studies from one centre was selected in the analysis. RESULTS: A total of 39 studies were identified, of which 25 met the inclusion criteria. The number of patients included in the meta-analysis was 998: 499 with non-enhanced MRA and 499 with gadolinium-enhanced MRA. The sensitivity and specificity of non-enhanced MRA were 94% (95% CI: 90-97%) and 85% (95% CI: 82 87%), respectively. For gadolinium-enhanced MRA sensitivity was 97% (95% CI: 93 98%) and specificity was 93% (95% CI: 91-95%). Thus, specificity and positive predictive value were significantly better for gadolinium-enhanced MRA (P < 0.001). Accessory renal arteries were depicted better by gadolinium-enhanced MRA (82%; 95% CI: 75-87%) than non-gadolinium MRA (49%; 95% CI: 42-60%) (P < 0.001). CONCLUSIONS: Gadolinium-enhanced MRA may replace arteriography in most patients with suspected renal artery stenosis, and has major advantages in that it is non invasive, avoids ionizing radiation and uses a non-nephrotoxic contrast agent. PMID- 12096863 TI - Prospective comparison of MRA with catheter angiography in the assessment of patients with aortoiliac occlusion before surgery or endovascular therapy. AB - AIM: The purpose of this study was to evaluate the usefulness of gadolinium enhanced 3D magnetic resonance (MR) angiography (CE MRA) as an alternative to translumbar or brachial angiography in the pre-operative work-up of patients with aortoiliac occlusion. MATERIALS AND METHODS: Nineteen patients (14 men and five women; age range 45-77 years; mean 62 years), not suitable for perfemoral angiography (aortoiliac occlusion, n = 18; infected femoro-femoral graft with femoral artery pseudoaneurysm, n = 1), underwent pre-operative CE MRA and catheter angiography (translumbar, n = 5; brachial, n = 14). CE MRA was performed using a 3D fast spoiled gradient-recalled pulse sequence during the intravenous injection of 40 ml of gadolinium DTPA and a 32-s breath-hold. All patients subsequently underwent surgical (n = 13) or percutaneous transluminal (n = 6) treatment for their vascular disease. The accuracy of CE MRA was determined compared with the findings at catheter angiography taken as the gold standard. RESULTS: CE MRA gave accurate information about the occlusion, inflow and distal run-off in the majority of patients. CE MRA revealed occlusions with an accuracy of 94.7% in the aortic segment, 98.7% in the iliac segment, and 100% in the common femoral segment. The arterial segments distal to the common femoral artery were not completely visualized in four patients but CE MRA provided sufficient information to plan either surgical or percutaneous transluminal therapy in all but one patient. CONCLUSION: CE MRA is highly accurate in showing the presence and extent of aortoiliac occlusions. In our study group, CE MRA gave sufficient information in the pre-operative evaluation of aortoiliac occlusion. PMID- 12096864 TI - Segmental lumbar spine instability at flexion-extension radiography can be predicted by conventional radiography. AB - AIM: To identify plain radiographic findings that predict segmental lumbar spine instability as shown by functional flexion-extension radiography. MATERIALS AND METHODS: Plain radiographs and flexion-extension radiographs of 215 patients with clinically suspected lumbar spine instability were analysed. Instability was classified into anterior or posterior sliding instability. The registered plain radiographic findings were traction spur, spondylarthrosis, arthrosis of facet joints, disc degeneration, retrolisthesis, degenerative spondylolisthesis, spondylolytic spondylolisthesis and vacuum phenomena. Factors reaching statistical significance in univariate analyses (P < 0.05) were included in stepwise multiple logistic regression analysis. RESULTS: Degenerative spondylolisthesis (P = 0.004 at L3-4 level and P = 0.017 at L4-5 level in univariate analysis and odds ratio 16.92 at L4-5 level in multiple logistic regression analyses) and spondylolytic spondylolisthesis (P = 0.003 at L5-S1 level in univariate analyses) were the strongest independent determinants of anterior sliding instability. Retrolisthesis (odds ratio 10.97), traction spur (odds ratio 4.45) and spondylarthrosis (odds ratio 3.20) at L3-4 level were statistically significant determinants of posterior sliding instability in multivariate analysis. CONCLUSION: Sliding instability is strongly associated with various plain radiographic findings. In mechanical back pain, functional flexion-extension radiographs should be limited to situations when symptoms are not explained by findings of plain radiographs and/or when they are likely to alter therapy. PMID- 12096865 TI - Comparison of mammographic and sonographic findings in typical and atypical medullary carcinomas of the breast. AB - AIM: The aim of this study was to describe the contribution of mammographic and sonographic findings to the discrimination of typical and atypical histopathologic groups of medullary carcinomas of the breast. MATERIALS AND METHODS: Imaging findings were retrospectively assessed in 33 women with medullary carcinomas (15 typical medullary carcinomas and 18 atypical medullary carcinomas) identified during pre-operative mammography. Twenty-nine of these women also had ultrasound and these findings were reviewed. RESULTS: Mammography showed a well circumscribed mass in 10 of the 15 (67%) typical medullary carcinomas and in four of the 17 (24%) atypical medullary carcinomas (P < 0.02). One small tumour in a woman with atypical medullary carcinoma was missed on mammography and was shown only on sonography. Sonographically, an irregular margin surrounding the whole mass or part of it was seen in three out of 14 (21%) patients with typical medullary carcinoma and in nine out of 15 (60%) patients with atypical medullary carcinomas (P < 0.05). Posterior acoustic shadowing was more often observed in the typical medullary carcinoma group than in atypical medullary carcinoma and the difference was found to be statistically significant (P < 0.05). None of the other mammographic and sonographic findings were sufficiently characteristic to allow for a differentiation between two groups. CONCLUSION: When typical medullary carcinomas were compared with atypical medullary carcinomas according to imaging features, they tended to be well circumscribed masses on both mammography and sonography, and a posterior acoustic shadow was not found on sonography. However, the imaging findings in these two subgroups often resembled each other and histopathology will always be required to confirm the diagnosis. PMID- 12096866 TI - Influence of therapy with iodine-131 on thyroid tissue pattern in colour and power Doppler sonography. AB - AIM: The aim of this study was to evaluate the influence of radioiodine therapy on sonographic thyroid patterns using power Doppler (PD) and colour Doppler (CD) sonography in hyperthyroid patients with autonomous nodules (AN) and Graves' disease (GD). METHOD: B-mode, colour, and power Doppler sonography, (99m)Tc scintigraphy, and laboratory analyses (free thyronine fT(3), free thyroxine fT(4), thyroid stimulating hormone TSH) were performed in 55 patients (AN = 27, GD = 28) before and 6 months following therapy with (131)I radioiodine therapy (RIT). RESULTS: In patients with an AN (but not in GD), a significant reduction in thyroid vascularization was subjectively noted following radioiodine therapy on both CD and PD ultrasound (Wilcoxon matched pairs, P < 0.05). The pre therapeutic grade of hypervascularization in the periphery of autonomous nodules correlated closely with the laboratory parameters of hyperthyroidism. As expected, PD indicated a higher grade of vascularization when compared with CD due to its greater sensitivity to flow. CONCLUSIONS: Radioiodine therapy led to a significant reduction in hypervascularization in patients with AN (but not in GD) corresponding to the normalization of serological values. Comparing CD and PD, PD detected a greater number of vessels. CD and PD are not able to replace scintigraphy and/or laboratory analyses in the management of patients with hyperthyroidism. PMID- 12096867 TI - Radiological considerations in the diagnosis of an endolymphatic sac tumour. PMID- 12096868 TI - Management of traumatic iliac vein pseudoaneurysm by transcatheter embolization. PMID- 12096869 TI - A congenital venous anomaly, with a portal-systemic shunt into a previously undescribed intra-thoracic vein. PMID- 12096871 TI - Phonological neighborhood effects in aphasic speech errors: spontaneous and structured contexts. AB - The current study investigates the influence of phonological neighborhoods on the accuracy of speech production in aphasia by examining errors produced in both spontaneous and structured speech tasks. Characteristics of the phonological neighborhoods of spontaneously produced aphasic errors are compared to the neighborhood characteristics of correctly produced targets in a picture description task. Accuracy of picture naming is also examined with reference to the phonological neighborhood characteristics of the stimuli. Results show that frequency of occurrence and neighborhood density play a facilitative role in speech production, replicating findings from recent studies with normal subjects. It is argued that the results are most parsimoniously explained within an interactive activation framework of lexical access. PMID- 12096872 TI - Semantic range and relevance of emotive utterances in patients with frontotemporal degeneration. AB - Patterns of aberrant language due to lateralized frontal brain lesions have been described. The present study investigated the consequences of lateralized lesion for aspects of semantic range and relevance in speech of patients with frontotemporal degeneration. Demented patients with predominantly left (n=10) and right (n=4) brain degeneration as well as 5 healthy controls participated. Significant differences were found concerning semantic range and relevance in descriptions of emotionally loaded pictures within the demented group and between the demented group and the controls. The demented individuals' speech was less varied and relevant in contrast to the healthy individuals. The speech of the left hemispheric group was more relevant but also more stereotypic and unspecified than that of the right hemispheric group. The results are discussed in terms of impaired semantic retrieval and in relation to lateralization of lesion. PMID- 12096873 TI - Predicability (ease of predication) as semantic substrate of imageability in reading and retrieval. AB - It was proposed by Jones (1985) that the apparent influence of a word's imageability upon the probability of the word being read correctly by a deep dyslexic person could be understood in terms of an underlying semantic variable, ease of predication (also termed predicability). In a recent critique, de Mornay Davies and Funnell (2000) claim to have identified a number of problems with the ease of predication proposal. It is shown here, however, that it is the critique itself which is fundamentally flawed. In contrast, the predicability approach continues to identify correctly the semantic substrate of apparent effects of imageability upon reading and memory retrieval. PMID- 12096874 TI - Role of the left hemisphere in sign language comprehension. AB - We investigated the relative role of the left versus right hemisphere in the comprehension of American Sign Language (ASL). Nineteen lifelong signers with unilateral brain lesions [11 left hemisphere damaged (LHD) and 8 right hemisphere damaged (RHD)] performed three tasks, an isolated single-sign comprehension task, a sentence-level comprehension task involving simple one-step commands, and a sentence-level comprehension task involving more complex multiclause/multistep commands. Eighteen of the participants were deaf, one RHD subject was hearing and bilingual (ASL and English). Performance was examined in relation to two factors: whether the lesion was in the right or left hemisphere and whether the temporal lobe was involved. The LHD group performed significantly worse than the RHD group on all three tasks, confirming left hemisphere dominance for sign language comprehension. The group with left temporal lobe involvement was significantly impaired on all tasks, whereas each of the other three groups performed at better than 95% correct on the single sign and simple sentence comprehension tasks, with performance falling off only on the complex sentence comprehension items. A comparison with previously published data suggests that the degree of difficulty exhibited by the deaf RHD group on the complex sentences is comparable to that observed in hearing RHD subjects. Based on these findings we hypothesize (i) that deaf and hearing individuals have a similar degree of lateralization of language comprehension processes and (ii) that language comprehension depends primarily on the integrity of the left temporal lobe. PMID- 12096875 TI - Knowledge of object manipulation and object function: dissociations in apraxic and nonapraxic subjects. AB - An influential account of selective semantic deficits posits that visual features are heavily weighted in the representations of animals, whereas information about function is central in the representations of tools (e.g., Warrington & Shallice, 1984 ). An alternative account proposes that information about all types of objects-animate and inanimate alike-is represented in a distributed semantic architecture by verbal-propositional, tactile, visual, and proprioceptive-motor nodes, reflecting the degree to which these systems were activated when the knowledge was acquired (e.g., Allport, 1985 ). We studied a group of left hemisphere chronic stroke patients, some of whom were apraxic, with measures of declarative tool and animal knowledge, body part knowledge, and function and manipulation knowledge of artifacts. Apraxic (n=7) and nonapraxic (n=6) subjects demonstrated a double dissociation of performance on tests of tool and animal knowledge, suggesting that the apraxic group was not simply more severely impaired overall. Apraxics were relatively impaired in manipulation knowledge, whereas nonapraxics tended to be relatively impaired in function knowledge. Apraxics were also more impaired with body parts than nonapraxics. The association of gestural praxis, tool knowledge, body part knowledge, and manipulation knowledge suggests a coherent basis for the organization of semantic artifact knowledge in frontoparietal cortical regions specialized for sensorimotor functions, and thus provides support for the distributed architecture account of the semantic system. PMID- 12096876 TI - Orthographic neighborhood size, number of word meanings, and number of higher frequency neighbors. AB - The present lexical decision task experiment examined orthographic (large, medium, small), number-of-meanings (ambiguous, unambiguous), and number of higher frequency neighbors (few, many), factors that have to date not been studied simultaneously. For both mean reaction time and percentage error, the critical three-way interaction between these factors was significant. Breakdown of this interaction revealed that the ambiguity effect (unambiguous-ambiguous) decreased as neighborhood size increased, but only when there were many higher frequency neighbors in the neighborhood. These results appear inconsistent with serial search models but are understandable within the context of interactive-activation models of word recognition. PMID- 12096877 TI - The contribution of the right cerebral hemisphere to the recovery from aphasia: a single longitudinal case study. AB - We examined the role of the right cerebral hemisphere in the recovery from aphasia of HJ, a 50-year-old right-handed and unilingual man who suffered from severe aphasia caused by an extensive left hemisphere (LH) lesion. He was followed-up over 10 months at 4-month intervals, with a lateralized lexical decision task (LDT), an attentional task, and a language battery. Testing started when HJ was 2 months poststroke. In the LDT, words were presented to central vision or lateralized to the left or right visual hemifield. At each test period, we examined the effect of the degree of imageability (high vs. low), and the grammatical class (noun vs. verb) of the targets on HJ's response times and error rates, with left visual field, right visual field, and central vision presentations. The results of the experiment showed that the pattern obtained with the LDT could not be accounted for by fluctuations in attention. There was an interaction of grammatical class with degree of imageability with left visual field displays only. The right hemisphere (RH) was faster with high-imageability words than with low-imageability words, regardless of their grammatical class. There was also an overall RH advantage on response times at 2 and 6 months after onset. This RH predominance coincided with a major recovery of language comprehension and the observation of semantic paralexias, while no major change in language expression was observed at that point. Ten months after onset, the pattern of lateralization changed, and response times for the LDT with either presentation site were equivalent. This LH improvement coincided with some recovery of language expression at the single-word level. The results of this study suggest that, in cases of severe aphasia caused by extensive LH lesions, the RH may play an important role in the recovery process. Furthermore, these results show that the contribution of the two cerebral hemispheres to recovery may vary overtime and affect specific aspects of language. PMID- 12096878 TI - Differences in language performance in variations of lateralization. AB - Language proficiency was studied in 489 primary-school-aged children classified as extreme left-, extreme right-, mild left-, and mild right-handers on the basis of both hand-preference and hand-skill, using a test battery of seven measures. An ANOVA run on factor scores showed neither significant association of language proficiency with variations of lateralization regarding hand-preference and hand skill nor differences in factor structure derived from a principal component analysis between extreme vs. mild hand-preference and hand-skill groups. Moreover, low language proficiency was not significantly associated with specific patterns of lateralization in hand-preference subgroups. In contrast, low language performers with poor hand-skill were significantly overrepresented both in the extreme left-handed group alone and when combined with the extreme right handed, in comparison to mild left- and mild right-handed with respect to population. The data are not consistently accommodated by the theory of balanced polymorphism (mainly Annett, 1978, 1985 Annett & Manning, 1989 ). Alternately, factors such as lag of maturation ( Bishop, 1980, 1984, 1990a, 1990b ), delay of growth ( Geschwind & Galaburda, 1985b ), and developmental instability associated with unique patterns of variations in lateralization ( Yeo, Gangestad, & Daniel, 1993 ) are discussed as possible factors accounting for the present results. PMID- 12096881 TI - Norepinephrine, the beta-adrenergic receptor, and immunity. AB - Over the past 20 years, a significant effort has been made to define a role for the neuroendocrine system in the regulation of immunity. It was expected that these experimental findings would help to establish a strategy for the development of clinical interventions to either suppress or augment immunological function for disease prevention. However, the translation of these basic experimental findings into clinical interventions has been difficult. Possible explanations for this difficulty are that the findings from human and animal studies do not agree and/or that the results obtained within one species are rarely verified in the other. Our goal in writing this review is to address this issue by summarizing the published findings from human studies and comparing them to published findings from animal studies. Although far from being exhaustive, this review summarizes and discusses at least the past 10 years of findings in which a change in immunity and a change in catecholamine levels and/or stimulation of the beta(2)-adrenergic receptor has been documented. PMID- 12096882 TI - Altered expression of autonomic neurotransmitter receptors and proliferative responses in lymphocytes from a chronic mild stress model of depression: effects of fluoxetine. AB - We studied beta-adrenergic and muscarinic cholinergic receptor (MR) expression and proliferative response in lymphocytes from animals under chronic mild stress (CMS) model of depression (CMS animals). Animals were subjected to CMS (periods of food or water deprivation, changes in lighting conditions, tilted cage, etc.) for 12 weeks. CMS lymphocytes showed an altered mitogen-induced proliferation. CMS-B and -T lymphocytes showed an increment on beta-adrenoceptor number and on intracellular responses to a beta-agonist. CMS-T cells showed higher MR expression and lower cGMP responses than normal lymphocytes. MR were not detectable in normal B cells while CMS-B cells showed both MR expression and cGMP response. Beta and muscarinic stimulation influenced lymphocyte proliferative responses, in accordance with cAMP and cGMP responses. After 12 weeks of the CMS procedure, animals were treated with fluoxetine while the CMS procedure continued. Fluoxetine treatment reverted the alterations induced by CMS. These findings suggest a possible mechanism for the immune alterations found in depressive disorders and for the effect of fluoxetine treatment on immune response. PMID- 12096883 TI - Changes in the leukocyte distribution and surface expression of adhesion molecules induced by hypothalamic stimulation in the cat. AB - Emotions and the neuroendocrine system are known to affect leukocyte distribution. However, there have so far been few reports on the relationship between hypothalamically induced emotional behavior and the endocrine-immune response. We previously reported changes in the leukocyte distribution and adhesion molecules induced by anteromedial hypothalamus stimulation (AH stimulation), which elicits restlessness behaviors in the cat. In this study, we examined ventromedial hypothalamus stimulation (VMH stimulation), which elicits threat behaviors. In addition, the endocrine responses after VMH stimulation were evaluated. VMH stimulation as well as AH stimulation induced elevations of plasma cortisol and epinephrine levels and granulocytosis and lymphopenia. In contrast, VMH stimulation induced only an elevation of plasma norepinephrine and elicited an opposite pattern of CD62L expression on the leukocyte subpopulations. The different endocrine-immunological reactions between VMH stimulation and AH stimulation were thus associated with different types of behavioral responses. PMID- 12096884 TI - Seasonal dependency of the effects of rotational stress and cyclophosphamide in mice bearing lewis lung carcinoma. AB - The antitumor effects of cyclophosphamide were previously shown to be markedly reduced by the application of restraint stress in mice bearing Lewis lung carcinoma. The aim of this work was to determine the effects of rotational stress on the antitumor action of cyclophosphamide in the same animal-tumor system. Since the effects of rotational stress on metastasis were found to display a circannual rhythm, with a maximum in summer and a minimum in winter, the experiments were performed in June and February. Groups of 10 young female mice were kept under low stress housing conditions, with a 12-12 h light/dark cycle, starting 2 weeks before and during each experiment. Rotational stress caused an increase of metastasis volume to 361% of nonstressed controls in June and a decrease to 32.4% in February. In both seasons, the treatment with cyclophosphamide (240 mg/kg/day for 6 days) caused the absence of detectable metastasis at sacrifice in all mice; its combination with rotational stress caused the presence of metastases in similar proportions (6/10 and 10/10 for June vs February, respectively). The survival time of control mice was approximately twice as long in February as in June and was not appreciably modified by rotational stress; cyclophosphamide was similarly active in both seasons (4/10 and 6/10 long-term survivors for June vs February, respectively), and the number of long-term survivors was reduced to 0/10 in both seasons by rotational stress. The survival of the different experimental groups inversely correlated with the number of metastases as determined at sacrifice at the end of treatment and also with the number of CD3(+) and CD4(+) splenic T-lymphocyte subsets. These results do not appear to depend on the disruption of the circadian organization of the mice by rotational stress or by seasonal differences in cyclophosphamide activity. On the other hand, they can be interpreted assuming that cyclophosphamide reduces tumor metastasis and that T-lymphocyte-mediated immune responses of the host, amenable to modulation by stress and displaying seasonal differences uncoupled from circadian rhythms, further contribute to the tumor inhibitory effects of the drug. The observed differences in tumor metastasis caused by rotational stress and survival time in two different seasons, and the marked attenuation of cyclophosphamide antitumor action by rotational stress, appear of interest for their experimental and clinical implications. PMID- 12096885 TI - Personality, brain asymmetry, and neuroendocrine reactivity in two immune mediated disorders: a preliminary report. AB - Development of some immune-mediated disorders may depend on dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis. To explore neuropsychologic mechanisms in relation to the abnormal endocrine reactivity in patients with systemic lupus erythematosus (SLE) and chronic hepatitis C (CHC) we used the corticotropin releasing hormone (CRH) test, the Minnesota Multiphasic Personality Inventory (MMPI), and the Edinburgh Inventory of Manual Preference Inventory (EIMP). Compared to controls, the adrenocorticotrophic hormone (ACTH) response to CRH was reduced in CHC, while SLE presented reduced baseline dehydroepiandrosterone sulfate levels; higher neurotic scores were found in SLE and higher behavior deviant scores in CHC. Peak ACTH levels were a significant factor for the MMPI profile variability, while the manual preference score was a significant factor for the ACTH response. Personality and manual preference contribute to neuroendocrine abnormalities. Different behavioral and neuroimmunoendocrine models emerge for these disorders. PMID- 12096886 TI - Effects of interleukin-1beta on food-maintained behavior in the mouse. AB - The present studies compared the effect of parenteral administration of the proinflammatory cytokine interleukin-1beta (IL-1beta) on food-seeking behavior under various conditions. IL-1beta (100 ng/mouse) decreased home cage consumption of sweetened milk to a greater extent in ad libitum fed mice than in mice that were food-restricted to maintain 85-90% of their free-feeding body weight. When operant responding for milk was maintained under a fixed-ratio 10 response (FR10) schedule of milk delivery, IL-1beta (30-300 ng/mouse) significantly decreased milk-maintained responding in mice fed ad libitum, but not in food-restricted mice. When food-restricted mice were trained under either an FR4 or FR32 response schedule of milk delivery, IL-1beta (100-300 ng/mouse) produced significant decreases in FR32, but not in FR4 responding. When responding was maintained under a progressive-ratio 10 response (PR10) schedule of milk delivery, IL-1beta (30-300 ng/mouse) dose-dependently decreased breaking points. These results indicate that the effects of IL-1beta on food-maintained behavior depend on both the level of motivation (as assessed by food restriction) and on the response cost for the milk (as assessed by ratio requirement). These findings suggest that motivational factors may be capable of attenuating some of the behavioral effects of these agents. PMID- 12096887 TI - The effects of acute psychological stress on lymphocyte adhesion molecule expression and density in cardiac versus vascular reactors. AB - This study examined the effects of acute psychological stress on lymphocyte subsets and their differential changes according to their cell adhesion molecule expression in cardiac versus vascular reactors. We classified 49 subjects into cardiac or vascular reactors based on the participants' cardiac output or total peripheral resistance reactivity to a speech presentation task. Analysis demonstrated that there were no significant differences in lymphocyte counts or adhesion molecule expression between cardiac and vascular reactors at rest. Cardiac reactors showed a significant decrease of surface density of CD62L on mixed lymphocytes (p <.001) as well as on CD4 (p <.01) and CD8 T-cells (p <.001). There was also a disproportionate increase in the number of CD62L(-) T cells compared to CD62L(+) T cells only in cardiac reactors (p <.001). There were no significant effects of the stressor observed in vascular responders. The findings replicate previous studies demonstrating associations between cardiovascular and immune responses to acute stress and extends those findings by suggesting that the relationship is more significant in individuals who increase their blood pressure primarily through a cardiac mechanism. PMID- 12096889 TI - Protective effect of prior acute immune challenge, but not footshock, on inflammation in the rat. AB - Previous studies have revealed that a single exposure to an acute stress or acute immune stimulus can produce long-lasting changes in the activity and responsiveness of the hypothalamo-pituitary-adrenal (HPA) axis. The HPA axis is believed to be an important component in determining the susceptibility and severity of inflammation in autoimmune disease models such as adjuvant-induced arthritis (AA). In the present study we have tested the hypothesis that a single exposure to either footshock or lipopolysaccharide (LPS) 3 weeks prior to adjuvant injection can alter susceptibility to AA. Changes in HPA axis parameters were also determined. The results demonstrated that prior exposure to LPS conferred resistance to inflammation in AA, which was not related to a delay in onset of inflammation but rather an alteration in susceptibility. In contrast, prior exposure to the acute stress of footshock did not alter susceptibility. HPA axis parameters were increased in adjuvant-injected rats whether inflammation was present or not. These data suggest that prior exposure to acute immune stimuli, but not to acute footshock stress, may alter susceptibility to inflammation in the rat AA model. These changes in susceptibility do not appear to be solely mediated by increases in HPA axis activity, which were apparent in all AA groups irrespective of the presence of inflammation. PMID- 12096888 TI - Postnatal lipopolysaccharide-induced illness predisposes to periodontal disease in adulthood. AB - The long-term consequences of neonatal lipopolysaccharide (LPS) exposure on adult behavioral and neuroendocrine stress responsiveness as well as on the clinical course of periodontal disease were assessed in male Lewis rats. At 3 and 5 days of age, pups were administered either saline (SHAM) or LPS or were left undisturbed. After postnatal treatment, mothers licked LPS-treated pups significantly more. In adult LPS rats of 3-5 months of age, home cage activity indicated changes of the diurnal rhythmicity. Furthermore, SHAM- and LPS-treated animals displayed treatment-specific signs of increased anxiety in social interaction, elevated plus maze, holeboard, and open field tests. At 7 months of age, a dramatic increase of periodontal fiber loss in LPS rats was associated with increased plasma interleukin-6 levels. In contrast, SHAM treatment caused high plasma interferon-gamma cytokine levels and protective effects in periodontal disease. Parameters of the response to novelty were significantly correlated with later disease susceptibility. Thus, LPS-induced early postnatal illness modulates the adult behavioral responsiveness to stress and predisposes to periodontal disease. PMID- 12096890 TI - Thymulin reduces the hyperalgesia and cytokine upregulation induced by cutaneous leishmaniasis in mice. AB - Cutaneous leishmaniasis (CL) in mice has been shown to produce hyperalgesia and upregulation of interleukin (IL)-1beta and nerve growth factor (NGF) levels. The aim of this study was to investigate the effects of thymulin on CL-induced hyperalgesia and cytokine upregulation. Daily treatment with thymulin (1, 100, and 1000 ng/ip) produced dose-dependent decreases in CL-induced hyperalgesia as assessed by the tail flick and the hot plate tests. The levels of NGF and IL 1beta were determined in the skin tissues of the hind leg in different groups (n = 5 each) of mice over a period of 5 weeks. Mice with CL showed sustained increase in the levels of IL-1beta and NGF which were reversed by thymulin (1 microg). Injection of thymulin only did not alter the nociceptive thresholds or the levels of IL-1beta and NGF. We conclude that thymulin can modulate the hyperalgesia induced by CL by decreasing the levels of the proinflammatory factors IL-1beta and NGF. PMID- 12096891 TI - Prior stressor exposure sensitizes LPS-induced cytokine production. AB - Exposure to stressors often alters the subsequent responsiveness of many systems. The present study tested whether prior exposure to inescapable tailshock (IS) alters the interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha, or IL-6 response to an injection of bacterial endotoxin (lipopolysaccharide; LPS). Rats were exposed to IS or remained as home cage controls (HCC); 24 h later animals were injected i.p. with either 10 microg/kg LPS or equilvolume sterile saline. IS significantly increased plasma TNF-alpha, IL-1beta, and pituitary, hypothalamus, hippocampus, cerebellum IL-1beta 1 h, but not 2 h, after LPS, compared to controls. Additional animals were injected with LPS or saline 4, 10, or 21 days after exposure to IS and tail vein blood was collected and assayed for IL-1beta. An enhanced plasma IL-1beta response occurred 4 days after IS, but was gone by 10 days. These results suggest that exposure to IS sensitizes the innate immune response to LPS by resulting in either a larger or a more rapid induction of proinflammatory cytokines. PMID- 12096892 TI - Neonatal lead exposure potentiates sickness behavior induced by Listeria monocytogenes infection of mice. AB - The effects of lead (Pb) administration on infection-induced decreases in water intake, food intake, and body weight gain have been assessed as manifestations of sickness behavior using a BALB/c mouse model. Pb acetate (0.5 mM) was administered via drinking water to dams from Day 0 postpartum to weaning and to mouse pups after weaning until sacrifice. At 22 days after birth, young mice were infected with Listeria monocytogenes. Mice with blood Pb levels of less than 25 microg/dl exhibited enhanced and prolonged sickness behavior compared to mice not exposed to Pb. With this mouse model, after infection, serum levels of interleukin (IL)-1beta and IL-6 were enhanced in Pb-exposed mice. Compared with control infected mice, significant reductions in the number of thymic CD4(+)CD8(+), CD4(+), CD8(+), and CD4(-)CD8(-) T cells were observed in Pb exposed mice. As a substitute for the infection, mice were injected with IL-1 and/or IL-6; Pb exacerbated sickness behavior only in mice injected peripherally with IL-1 and IL-6. Our data in young mice suggest that children with blood Pb levels during bacterial infection may exhibit enhanced and prolonged sickness behavior due to Pb/cytokine-dependent processes and that Pb appears to influence sickness behavior depending on the types and amounts of cytokines generated. PMID- 12096893 TI - Phenylethanolamine N-methyl transferase expression in mouse thymus and spleen. AB - Catecholamines usually are found in neurons and chromaffin cells of mammals. In this study, surprisingly high levels of the epinephrine synthesizing enzyme phenylethanolamine N-methyl transferase (PNMT) were detected in the thymus of young mice. Levels of PNMT activity in the thymus were comparable to levels in the brainstem and were suppressed by the PNMT inhibitor LY134046. PNMT mRNA was localized with in situ hybridization throughout the thymus, but levels were approximately twofold higher in the cortex than in the medulla. PNMT activity was barely detectable in the spleen, and only a few cells expressing PNMT mRNA were located in the marginal zone of the white pulp. These findings suggest that cells in the thymus of young mice have the ability to synthesize epinephrine. PMID- 12096894 TI - Symmetry embedding in the reconstruction of macromolecular assemblies via the discrete Radon transform. AB - In this paper we discuss the embedding of symmetry information in an algorithm for three-dimensional reconstruction, which is based on the discrete Radon transform. The original algorithm was designed for randomly oriented and in principal asymmetric particles. The expanded version presented here covers all symmetry point groups which can be exhibited by macromolecular protein assemblies. The orientations of all symmetry equivalent projections, based on the orientation of an experimental projection, are obtained using global group operators. Further, an improved interpolation scheme for the recovery of the three-dimensional discrete Radon transform has been designed for greater computational efficiency. The algorithm has been tested on phantom structures as well as on real data, a virus structure possessing icosahedral symmetry. PMID- 12096895 TI - A relational database for cryoEM: experience at one year and 50 000 images. AB - For the past year we have been using a relational database as part of an automated data collection system for cryoEM. The database is vital for keeping track of the very large number of images collected and analyzed by the automated system and essential for quantitatively evaluating the utility of methods and algorithms used in the data collection. The database can be accessed using a variety of tools including specially developed Web-based interfaces that enable a user to annotate and categorize images using a Web-based form. PMID- 12096896 TI - Single molecule imaging of supported planar lipid bilayer--reconstituted human insulin receptors by in situ scanning probe microscopy. AB - A 480-kDa disulfide-linked heterodimer single-pass transmembrane protein, the insulin receptor, is autophosphorylated upon insulin binding to its extracellular domain. Remarkably, the structural basis for this activation process remained largely unknown until the recent cryoelectron microscopy studies of the insulin insulin receptor complex by Luo et al. [Science 285 (1999) 1077]. We report here the results of an in situ study by high-resolution scanning probe microscopy of the full-length insulin receptor reconstituted within supported planar lipid bilayers. Our preliminary studies confirm that (1) the intact receptor can be reconstituted constitutively within a lipid vesicle and (2) fusion of the receptor-containing vesicles to mica resulted in the formation of molecular flat 5.5-nm-thick supported planar bilayers populated by two populations of protrusions, the shape and size of which are consistent with those of the insulin receptor's intra- and extracellular domains as modeled by the cryo-EM data of Ottensmeyer et al. [Biochemistry 39 (2000) 12103]. These results establish a framework for real-time studies of insulin-insulin receptor binding by in situ SPM and single molecule force spectroscopy. PMID- 12096897 TI - IMIRS: a high-resolution 3D reconstruction package integrated with a relational image database. AB - Recent advances in electron cryomicroscopy instrumentation and single particle reconstruction have created opportunities for high-throughput and high-resolution three-dimensional (3D) structure determination of macromolecular complexes. However, it has become impractical and inefficient to rely on conventional text file data management and command-line programs to organize and process the increasing numbers of image data required in high-resolution studies. Here, we present a distributed relational database for managing complex datasets and its integration into our high-resolution software package IMIRS (Image Management and Icosahedral Reconstruction System). IMIRS consists of a complete set of modular programs for icosahedral reconstruction organized under a graphical user interface and provides options for user-friendly, step-by-step data processing as well as automatic reconstruction. We show that the integration of data management with processing in IMIRS automates the tedious tasks of data management, enables data coherence, and facilitates information sharing in a distributed computer and user environment without significantly increasing the time of program execution. We demonstrate the applicability of IMIRS in icosahedral reconstruction toward high resolution by using it to obtain an 8-A 3D structure of an intermediate sized dsRNA virus. PMID- 12096898 TI - Pure F-actin networks are distorted and branched by steps in the critical-point drying method. AB - Elucidation of the ultrastructural organization of actin networks is crucial for understanding the molecular mechanisms underlying actin-based motility. Results obtained from cytoskeletons and actin comets prepared by the critical-point procedure, followed by rotary shadowing, support recent models incorporating actin filament branching as a main feature of lamellipodia and pathogen propulsion. Since actin branches were not evident in earlier images obtained by negative staining, we explored how these differences arise. Accordingly, we have followed the structural fate of dense networks of pure actin filaments subjected to steps of the critical-point drying protocol. The filament networks have been visualized in parallel by both cryo-electron microscopy and negative staining. Our results demonstrate the selective creation of branches and other artificial structures in pure F-actin networks by the critical-point procedure and challenge the reliability of this method for preserving the detailed organization of actin assemblies that drive motility. PMID- 12096899 TI - Colocalization of heparin and histamine in the intracellular granules of test cells from the invertebrate Styela plicata (Chordata-Tunicata). AB - In most ascidian species the oocytes are surrounded by two types of accessory cells called follicle cells and test cells. Test cells are located on the periphery of oocytes and remain in the perivitelline space during egg development until hatching. Heparin and histamine were previously described in the test cells of the ascidian Styela plicata. In the present study, electron microscopy techniques were used to characterize the ultrastructure of the S. plicata test cells and to localize heparin and histamine in these cells. Test cells contain several intracellular granules with unique ultrastructural features. They are formed by elongated filaments composed of serial globules with an electron-lucent circle, containing a central electron-dense spot. Immunocytochemistry showed that heparin and histamine colocalize at the border of granule filaments in the test cell. Compound 48/80, a potent secretagogue of heparin-containing mast cells, also induced degranulation of test cells. According to these results, we suggest that test cells represent ancient effector cells of the innate immunity in primitive chordates. PMID- 12096900 TI - Importance of detergent and phospholipid in the crystallization of the human erythrocyte anion-exchanger membrane domain. AB - Three-dimensional crystals were obtained for the membrane domain of the human erythrocyte anion exchanger (AE1, Band 3). Protein homogeneity and stability and the delicate balance between the detergent used and the amount of phospholipids copurifying are critical to the formation of three-dimensional crystals of the AE1 membrane domain. While deglycosylation improved the protein homogeneity, its stability was significantly increased by inhibitor binding. Size-exclusion chromatography showed that the protein was monodisperse in detergents with acyl chains of 10-12 carbons over a pH range of 5.5-10.0. This pH range and the detergents that retained the protein's monodispersity were used for crystallization screening. Crystals were obtained with the protein purified in C(12)E(8), dodecylmaltoside, decylthiomaltoside, and cyclohexyl-hexylmaltoside. Five to 13 lipid molecules per protein were required for the protein crystal formation. Those crystals grown in dodecylmaltoside diffracted X-rays to 14 A. With these factors taken into consideration, ways to further improve the crystal quality are suggested. PMID- 12096901 TI - Characterisation of the BRCT domains of the breast cancer susceptibility gene product BRCA1. AB - The breast cancer susceptibility gene product BRCA1 is a tumour suppressor but the biochemical and biological functions that underlie its role in carcinogenesis remain to be determined. Here, we characterise the solution properties of the highly conserved C terminus of BRCA1, consisting of a tandem repeat of the BRCT domain (BRCT-tan), that plays a critical role in BRCA1-mediated tumour suppression. The overall free energy of unfolding of BRCT-tan is high (14.2 kcal mol(-1) at 20 degrees C in water) but unfolding occurs via an aggregation-prone, partly folded intermediate. A representative set of cancer-associated sequence variants was constructed and the effects on protein stability were measured. All of the mutations were highly destabilising and they would be expected to cause loss of function for this reason. Over half could not be purified in a soluble form, indicating that these residues are critical for maintaining structural integrity. The remaining mutants exhibited much greater aggregation propensities than the wild-type, which is most likely a consequence of their reduced thermodynamic stability relative to the partly folded intermediate. The mutations characterised here are located at different sites in the BRCT-tan structure that do not explain fully their effects on the protein's stability. Thus, the results indicate an important role for biophysical studies in assessing the significance of sequence variants and in determining how they cause disease. PMID- 12096902 TI - Identification of the catalytic subdomain of the VS ribozyme and evidence for remarkable sequence tolerance in the active site loop. AB - We show here that the ribozyme domain of the Neurospora VS ribozyme consists of separable upper and lower subdomains. Deletion analysis demonstrates that the entire upper subdomain (helices III/IV/V) is dispensable for site-specific cleavage activity, providing experimental evidence that the active site is contained within the lower subdomain and within the substrate itself. We demonstrate an important role in cleavage activity for a region of helix VI called the 730 loop. Surprisingly, several loop sequences, sizes, and structures at this position can support site-specific cleavage, suggesting that a variety of non-Watson-Crick structures, rather than a specific loop structure, in this region of the ribozyme can contribute to formation of the active site. PMID- 12096903 TI - Sequence and structural conservation in RNA ribose zippers. AB - The "ribose zipper", an important element of RNA tertiary structure, is characterized by consecutive hydrogen-bonding interactions between ribose 2' hydroxyls from different regions of an RNA chain or between RNA chains. These tertiary contacts have previously been observed to also involve base-backbone and base-base interactions (A-minor type). We searched for ribose zipper tertiary interactions in the crystal structures of the large ribosomal subunit RNAs of Haloarcula marismortui and Deinococcus radiodurans, and the small ribosomal subunit RNA of Thermus thermophilus and identified a total of 97 ribose zippers. Of these, 20 were found in T. thermophilus 16 S rRNA, 44 in H. marismortui 23 S rRNA (plus 2 bridging 5 S and 23 S rRNAs) and 30 in D. radiodurans 23 S rRNA (plus 1 bridging 5 S and 23 S rRNAs). These were analyzed in terms of sequence conservation, structural conservation and stability, location in secondary structure, and phylogenetic conservation. Eleven types of ribose zippers were defined based on ribose-base interactions. Of these 11, seven were observed in the ribosomal RNAs. The most common of these is the canonical ribose zipper, originally observed in the P4-P6 group I intron fragment. All ribose zippers were formed by antiparallel chain interactions and only a single example extended beyond two residues, forming an overlapping ribose zipper of three consecutive residues near the small subunit A-site. Almost all ribose zippers link stem (Watson-Crick duplex) or stem-like (base-paired), with loop (external, internal, or junction) chain segments. About two-thirds of the observed ribose zippers interact with ribosomal proteins. Most of these ribosomal proteins bridge the ribose zipper chain segments with basic amino acid residues hydrogen bonding to the RNA backbone. Proteins involved in crucial ribosome function and in early stages of ribosomal assembly also stabilize ribose zipper interactions. All ribose zippers show strong sequence conservation both within these three ribosomal RNA structures and in a large database of aligned prokaryotic sequences. The physical basis of the sequence conservation is stacked base triples formed between consecutive base-pairs on the stem or stem-like segment with bases (often adenines) from the loop-side segment. These triples have previously been characterized as Type I and Type II A-minor motifs and are stabilized by base-base and base-ribose hydrogen bonds. The sequence and structure conservation of ribose zippers can be directly used in tertiary structure prediction and may have applications in molecular modeling and design. PMID- 12096904 TI - Nucleolin associates with a subset of the human Ro ribonucleoprotein complexes. AB - Ro RNPs are evolutionarily conserved, small cytoplasmic RNA-protein complexes with an unknown function. In human cells, Ro RNPs consist of one of the four hY RNAs and two core proteins: Ro60 and La. Recently, the association of hnRNP I and hnRNP K with particles containing hY1 and hY3 RNAs has been described. The association of three other proteins, namely calreticulin, Ro52 and RoBPI, with (subsets of) the Ro RNPs is still controversial. To gain more insight into the composition and function of the Ro RNPs, we have immunopurified these particles from HeLa cell extracts using monoclonal antibodies against Ro60 and La. Using this approach, we have identified the RNA-binding protein nucleolin as a novel subunit of Ro RNP particles containing hY1 or hY3 RNA, but not hY4 and hY5 RNA. Using an in vitro hY RNA-binding assay we established that the internal pyrimidine-rich loop of hY1 and hY3 RNA is essential for the association of nucleolin. The binding is critically dependent on the presence of all four RNP motifs of nucleolin, but not of the C-terminal RGG-box. Moreover, we demonstrate that, in contrast to nucleolin and hnRNP K, nucleolin and hnRNP I can bind simultaneously to the internal pyrimidine-rich loop of hY1 RNA. We postulate that nucleolin functions in the biogenesis and/or trafficking of hY1 and hY3 RNPs through the nucleolus and subsequent transport to the cytoplasm. PMID- 12096905 TI - RNA hairpin invasion and ribosome elongation arrest by mixed base PNA oligomer. AB - Recently, we have shown that peptide nucleic acid (PNA) tridecamers targeted to the codon 74, 128 and 149 regions of Ha-ras mRNA arrested translation elongation in vitro. Our data demonstrated for the first time that PNAs with mixed base sequence targeted to the coding region of a messenger RNA could arrest the translation machinery and polypeptide chain elongation. The peculiarity of the complexes formed with PNA tridecamers and Ha-ras mRNA rests upon the stability of PNA-mRNA hybrids, which are not dissociated by cellular proteins or multiple denaturing conditions. In the present study, we show that shorter PNAs such as a dodecamer or an undecamer targeted to the codon 74 region arrest translation elongation in vitro. The 13, 12, and 11-mer PNAs contain eight and the 10-mer PNA seven contiguous pyrimidine residues. Upon binding with parallel Hoogsteen base pairing to the PNA-RNA duplex, six of the cytosine bases and one thymine base of a second PNA can form C.G*C(+) and T.A*T triplets. Melting experiments show two well-resolved transitions corresponding to the dissociation of the third strand from the core duplex and to melting of duplex at higher temperature. The enzymatic structure mapping of a target 27-mer RNA revealed a hairpin structure that is disrupted upon binding of tri-, dodeca-, undeca- and decamer PNAs. We show that the non-bonded nucleobase overhangs on the RNA stabilize the PNA-RNA hybrids and probably assist the PNA in overcoming the stable secondary structure of the RNA target. The great stability of PNA-RNA duplex and triplex structures allowed us to identify both 1:1 and 2:1 PNA-RNA complexes using matrix-assisted laser desorption/ionization time-of -flight mass spectrometry. Therefore, it is possible to successfully target mixed sequences in structured regions of messenger RNA with short PNA oligonucleotides that form duplex and triplex structures that can arrest elongating ribosomes. PMID- 12096906 TI - Binding of specific DNA base-pair mismatches by N-methylpurine-DNA glycosylase and its implication in initial damage recognition. AB - Most DNA glycosylases including N-methylpurine-DNA glycosylase (MPG), which initiate DNA base excision repair, have a wide substrate range of damaged or altered bases in duplex DNA. In contrast, uracil-DNA glycosylase (UDG) is specific for uracil and excises it from both single-stranded and duplex DNAs. Here we show by DNA footprinting analysis that MPG, but not UDG, bound to base pair mismatches especially to less stable pyrimidine-pyrimidine pairs, without catalyzing detectable base cleavage. Thermal denaturation studies of these normal and damaged (e.g. 1,N(6)-ethenoadenine, varepsilonA) base mispairs indicate that duplex instability rather than exact fit of the flipped out damaged base in the catalytic pocket is a major determinant in the initial recognition of damage by MPG. Finally, based on our determination of binding affinity and catalytic efficiency we conclude that the initial recognition of substrate base lesions by MPG is dependent on the ease of flipping of the base from unstable pairs to a flexible catalytic pocket. PMID- 12096907 TI - Symmetric DNA sites are functionally asymmetric within Flp and Cre site-specific DNA recombination synapses. AB - Flp and Cre-mediated recombination on symmetrized FRT and loxP sites, respectively, in circular plasmid substrates yield both DNA inversion and deletion. However, upon sequestering three negative supercoils outside the recombination complex using the resII-resIII synapse formed by Tn3 resolvase and the LER synapse formed by phage Mu transposase in the case of Flp and Cre, respectively, the reactions are channeled towards inversion at the expense of deletion. The inversion product is a trefoil, its unique topology being conferred by the external resolvase or LER synapse. Thus, Flp and Cre assign their symmetrized substrates a strictly antiparallel orientation with respect to strand cleavage and exchange. These conclusions are supported by the product profiles from tethered parallel and antiparallel native FRT sites in dilution and competition assays. Furthermore, the observed recombination bias favoring deletion over inversion in a nicked circular substrate containing two symmetrized FRT sites is consistent with the predictions from Monte Carlo simulations based on antiparallel synapsis of the DNA partners. PMID- 12096908 TI - Elucidating a key intermediate in homologous DNA strand exchange: structural characterization of the RecA-triple-stranded DNA complex using fluorescence resonance energy transfer. AB - The RecA protein of Escherichia coli plays essential roles in homologous recombination and restarting stalled DNA replication forks. In vitro, the protein mediates DNA strand exchange between single-stranded (ssDNA) and homologous double-stranded DNA (dsDNA) molecules that serves as a model system for the in vivo processes. To date, no high-resolution structure of the key intermediate, comprised of three DNA strands simultaneously bound to a RecA filament (RecA tsDNA complex), has been reported. We present a systematic characterization of the helical geometries of the three DNA strands of the RecA-tsDNA complex using fluorescence resonance energy transfer (FRET) under physiologically relevant solution conditions. FRET donor and acceptor dyes were used to label different DNA strands, and the interfluorophore distances were inferred from energy transfer efficiencies measured as a function of the base-pair separation between the two dyes. The energy transfer efficiencies were first measured on a control RecA-dsDNA complex, and the calculated helical parameters (h approximately 5 A, Omega(h) approximately 20 degrees ) were consistent with structural conclusions derived from electron microscopy (EM) and other classic biochemical methods. Measurements of the helical parameters for the RecA-tsDNA complex revealed that all three DNA strands adopt extended and unwound conformations similar to those of RecA-bound dsDNA. The structural data are consistent with the hypothesis that this complex is a late, post-strand-exchange intermediate with the outgoing strand shifted by about three base-pairs with respect to its registry with the incoming and complementary strands. Furthermore, the bases of the incoming and complementary strands are displaced away from the helix axis toward the minor groove of the heteroduplex, and the bases of the outgoing strand lie in the major groove of the heteroduplex. We present a model for the strand exchange intermediate in which homologous contacts preceding strand exchange arise in the minor groove of the substrate dsDNA. PMID- 12096909 TI - Esterolytic antibodies as mechanistic and structural models of hydrolases-a quantitative analysis. AB - Understanding enzymes quantitatively and mimicking their remarkable catalytic efficiency is a paramount challenge. Here, we applied esterolytic antibodies (the D-Abs) to dissect and quantify individual elements of enzymatic catalysis such as transition state (TS) stabilization, nucleophilic reactivity and conformational changes. Kinetic and mutagenic analysis of the D-Abs were combined with existing structural evidence to show that catalysis by the D-Abs is driven primarily by stabilization of the tetrahedral oxyanionic intermediate of ester hydrolysis formed by the nucleophilic attack of an exogenous (solution) hydroxide anion. The side-chain of TyrH100d is shown to be the main H-bond donor of the D-Abs oxyanion hole. The pH-rate and pH-binding profiles indicate that the strength of this H bond increases dramatically as the neutral substrate develops into the oxyanionic TS, resulting in TS stabilization of 5-7 kcal/mol, which is comparable to oxyanionic TS stabilization in serine hydrolases. We show that the rate of the exogenous (intermolecular) nucleophilic attack can be enhanced by 2000-fold by replacing the hydroxide nucleophile with peroxide, an alpha-nucleophile that is much more reactive than hydroxide. In the presence of peroxide, the rate saturates (k(cat)(max)) at 6 s(-1). This rate-ceiling appears to be dictated by the rate of the induced-fit conformational rearrangement leading to the active antibody-TS complex. The selective usage of negatively charged exogenous nucleophiles by the D-Abs led to the identification of a positively charged channel. Imprinted by the negatively-charged TS-analogue against which these antibodies were elicited, this channel presumably directs the nucleophile to the antibody-bound substrate. Our findings are discussed in comparison with serine esterases and, in particular, with cocaine esterase (cocE), which possesses a tyrosine based oxyanion hole. PMID- 12096910 TI - Crystal structure of the Ly49I natural killer cell receptor reveals variability in dimerization mode within the Ly49 family. AB - Natural killer (NK) cells play a crucial role in the detection and destruction of virally infected and tumor cells during innate immune responses. The cytolytic activity of NK cells is regulated through a balance of inhibitory and stimulatory signals delivered by NK receptors that recognize classical major histocompatabilty complex class I (MHC-I) molecules, or MHC-I homologs such as MICA, on target cells. The Ly49 family of NK receptors (Ly49A through W), which includes both inhibitory and activating receptors, are homodimeric type II transmembrane glycoproteins, with each subunit composed of a C-type lectin-like domain tethered to the membrane by a stalk region. We have determined the crystal structure, at 3.0 A resolution, of the murine inhibitory NK receptor Ly49I. The Ly49I monomer adopts a fold similar to that of other C-type lectin-like NK receptors, including Ly49A, NKG2D and CD69. However, the Ly49I monomers associate in a manner distinct from that of these other NK receptors, forming a more open dimer. As a result, the putative MHC-binding surfaces of the Ly49I dimer are spatially more distant than the corresponding surfaces of Ly49A or NKG2D. These structural differences probably reflect the fundamentally different ways in which Ly49 and NKG2D receptors recognize their respective ligands: whereas the single MICA binding site of NKG2D is formed by the precise juxtaposition of two monomers, each Ly49 monomer contains an independent binding site for MHC-I. Hence, the structural constraints on dimerization geometry may be relatively relaxed within the Ly49 family. Such variability may enable certain Ly49 receptors, like Ly49I, to bind MHC-I molecules bivalently, thereby stabilizing receptor-ligand interactions and enhancing signal transmission to the NK cell. PMID- 12096911 TI - The crystal structure and catalytic mechanism of cellobiohydrolase CelS, the major enzymatic component of the Clostridium thermocellum Cellulosome. AB - Cellobiohydrolase CelS plays an important role in the cellulosome, an active cellulase system produced by the thermophilic anaerobe Clostridium thermocellum. The structures of the catalytic domain of CelS in complex with substrate (cellohexaose) and product (cellobiose) were determined at 2.5 and 2.4 A resolution, respectively. The protein folds into an (alpha/alpha)(6) barrel with a tunnel-shaped substrate-binding region. The conformation of the loops defining the tunnel is intrinsically stable in the absence of substrate, suggesting a model to account for the processive mode of action of family 48 cellobiohydrolases. Structural comparisons with other (alpha/alpha)(6) barrel glycosidases indicate that CelS and endoglucanase CelA, a sequence-unrelated family 8 glycosidase with a groove-shaped substrate-binding region, use the same catalytic machinery to hydrolyze the glycosidic linkage, despite a low sequence similarity and a different endo/exo mode of action. A remarkable feature of the mechanism is the absence, from CelS, of a carboxylic group acting as the base catalyst. The nearly identical arrangement of substrate and functionally important residues in the two active sites strongly suggests an evolutionary relationship between the cellobiohydrolase and endoglucanase families, which can therefore be classified into a new clan of glycoside hydrolases. PMID- 12096912 TI - On the role of the crystal environment in determining protein side-chain conformations. AB - The role of crystal packing in determining the observed conformations of amino acid side-chains in protein crystals is investigated by (1) analysis of a database of proteins that have been crystallized in different unit cells (space group or unit cell dimensions) and (2) theoretical predictions of side-chain conformations with the crystal environment explicitly represented. Both of these approaches indicate that the crystal environment plays an important role in determining the conformations of polar side-chains on the surfaces of proteins. Inclusion of the crystal environment permits a more sensitive measurement of the achievable accuracy of side-chain prediction programs, when validating against structures obtained by X-ray crystallography. Our side-chain prediction program uses an all-atom force field and a Generalized Born model of solvation and is thus capable of modeling simple packing effects (i.e. van der Waals interactions), electrostatic effects, and desolvation, which are all important mechanisms by which the crystal environment impacts observed side-chain conformations. Our results are also relevant to the understanding of changes in side-chain conformation that may result from ligand docking and protein-protein association, insofar as the results reveal how side-chain conformations change in response to their local environment. PMID- 12096913 TI - Equilibrium and pressure-jump relaxation studies of the conformational transitions of P13MTCP1. AB - The conformational transitions of a small oncogene product, p13(MTCP1), have been studied by high-pressure fluorescence of the intrinsic tryptophan emission and high-pressure 1D and 2D 1H-15N NMR. While the unfolding transition monitored by fluorescence is cooperative, two kinds of NMR spectral changes were observed, depending on the pressure range. Below approximately 200 MPa, pressure caused continuous, non-linear shifts of many of the 15N and 1H signals, suggesting the presence of an alternate folded conformer(s) in rapid equilibrium (tau<80% success rate. CONCLUSIONS: This study shows that not only is the number of procedures important to outcome, but also the frequency with which they are performed. Based on our team's performance data, 55 cases would appear to be the minimum volume and 1 case every 10 days the minimum frequency to obtain good operative results with aortic endografting. PMID- 12096939 TI - The first 150 endovascular AAA repairs at a single institution: how steep is the learning curve? AB - PURPOSE: To determine whether increasing experience with endovascular abdominal aortic aneurysm (AAA) repair in a single institution will result in improved outcome. METHODS: A retrospective review was undertaken of 150 consecutive cases of endovascular AAA repairs performed using the AneuRx device between October 1996 and April 2000 in a university-based medical center. The population was divided into early and late groups of 75 patients each. Endpoints included technical success; complications; early (< or =30-day) morbidity, mortality and rupture; endoleak at discharge and at 1 month; early secondary intervention; proximal neck and iliac tortuosity; extender cuff placement; femoral reconstructions beyond primary repair; total fluoroscopy time; and contrast load. RESULTS: Baseline patient and aneurysm characteristics were similar between the 2 groups. Technical success was 98.7%; 2 cases were converted intraprocedurally owing to difficult iliac access (early group) and a severely angulated proximal neck (late group). There was a tendency toward more frequent use of intraoperative proximal extender cuffs in the early group (12% versus 4% in the late group, p=0.13). Femoral reconstructions were more frequent in the early group (36% versus 19%, p<0.025). While total contrast volume was similar (111 +/- 56 versus 105 +/- 45 mL, p=NS), total fluoroscopy time was significantly reduced (p<0.05) between the early and late groups. CONCLUSION: With attention to detail and careful patient selection, successful endovascular AAA repair can be achieved with very few conversions and low perioperative mortality even during the center's early experience. Evidence indicates, however, that a learning curve definitely exists, as shown by fewer access site problems, more accurate device deployments, and decreased fluoroscopy times as proficiency is attained. PMID- 12096935 TI - Newer antiepileptic drugs in bipolar disorder: rationale for use and role in therapy. AB - Antiepileptic drugs (AEDS) are used regularly in the treatment of patients with bipolar disorders. Carbamazepine and valproic acid (sodium valproate) are effective as antimanic treatments, and the success of these medications has prompted investigation of other AEDs as possible treatments in patients with mood disorders. Lamotrigine appears to be the most promising of the newer AEDs with respect to effects in mood disorders. Current evidence suggests efficacy of this drug both as monotherapy and as an adjunctive agent in bipolar depression, and studies are underway to clarify its efficacy in mood stabilisation and rapid cycling, as currently available data are equivocal. Use of gabapentin is not as well supported in the literature, although data from open trials using it as an adjunctive agent suggest that it may be helpful in patients with bipolar depression. There have been some open trials and case reports supporting the use of topiramate as an adjunctive agent for the treatment of mania; however, data from controlled trials are not yet available. Further controlled trials of lamotrigine, gabapentin or topiramate as monotherapy and adjunctive treatment are needed to clarify their potential roles in the treatment of patients with mood disorders. PMID- 12096940 TI - Endovascular repair of inflammatory abdominal aortic aneurysms. AB - PURPOSE: To report a single-center experience with endovascular repair of inflammatory abdominal aortic aneurysm (IAAA), with particular attention to the fate of the aneurysm sac, perianeurysmal fibrosis (PAF), and renal function. METHODS: A retrospective review of 350 patients undergoing endovascular aortic aneurysm repair during a 7-year period at University Hospital, Nottingham, identified 14 (4%) cases of IAAA confirmed either on preoperative spiral computed tomography (CT) or at laparotomy in attempted open aneurysm repair. All data were reviewed from a prospectively maintained database, hospital notes, and serial CT studies. RESULTS: Endovascular repair was successfully completed in all 14 IAAA patients, but 2 (14%) died in the perioperative period. One patient referred from another center was lost to imaging follow-up, leaving 11 patients who were followed for a mean 29 months (range 1-73). All 11 IAAAs remained excluded, but 1 patient required a secondary transabdominal intervention for a type III endoleak. There was no CT evidence of PAF progression in any patient. Postoperative renal complications were not encountered where there had been none preoperatively. CONCLUSIONS: IAAA may be successfully excluded by the endovascular technique, and EVAR is particularly useful where open repair has failed. The impact of endograft placement on perianeurysmal fibrosis is less clear. In this study, there was no suggestion that the degree of PAF worsens following endovascular repair. PMID- 12096941 TI - Institutional economic losses associated with AAA repair are independent of technique. AB - PURPOSE: To investigate the perception of significant economic loss associated with endovascular abdominal aortic aneurysm (AAA) repair by comparing economic variables for the open and endovascular techniques. METHODS: In a 1-year period, 20 consecutive patients (19 men; mean age 73.3 years, range 62-89) were treated for uncomplicated infrarenal AAAs using conventional open repair in 11 and endovascular repair (EVR) in 9. For the open repair, standard techniques were employed, including transperitoneal and retroperitoneal exposures; in EVR, both the AneuRx and Ancure systems were utilized. Length of stay and institutional costs were carefully tracked and compared. RESULTS: The patients were similar with regard to comorbidities, but the endograft patients were older (p=0.02) Length of stay was significantly lower in the EVR group (1.9 +/- 0.9 days) compared with the open group (8.4 +/- 4.5 days, p=0.0004). However, total mean treatment costs (open: $17,576 +/- $11,025 and EVR: $20,247 +/- $5003; p=0.51) and subsequent losses (open: -$3949 +/- $7095 and EVR: -$7572 +/- $4488; p=0.20) were not significantly different between the groups. CONCLUSIONS; The costs associated with the care of AAA patients are independent of the technique used for repair. The economic loss associated with treatment is directly related to inadequate reimbursement on the part of Medicare and other carriers. PMID- 12096942 TI - Predicting spinal cord ischemia before endovascular thoracoabdominal aneurysm repair: monitoring somatosensory evoked potentials. AB - PURPOSE: To present a method of predicting spinal cord ischemia before deployment of an endograft in the thoracoabdominal aorta. CASE REPORT: A 76-year-old high risk patient presented with an aneurysm of the distal thoracic and proximal abdominal aorta. An endovascular treatment was scheduled, but before deployment of the endograft, occlusion of the aneurysmal segment was monitored for 15 minutes using multilevel somatosensory evoked potentials (SEP). An external axillofemoral bypass was necessary during the test period to prevent distal ischemia; transesophageal echocardiography verified the absence of flow inside the aneurysm during occlusion. Because no SEP changes occurred during the 15 minute test, a 20-cm-long endograft was deployed. No postoperative neurological event was encountered, and the aneurysm has remained successfully excluded with shrinkage of the sac diameter by 1 cm at 6 months. CONCLUSIONS: An occlusion test to detect spinal cord ischemia before deployment of an endograft could be useful in lowering the risk of paraplegia associated with endovascular treatment of thoracoabdominal aneurysm. PMID- 12096943 TI - Endovascular exclusion of saccular AAAs using "stacked" AneuRx aortic cuffs. AB - PURPOSE: To show that AneuRx aortic cuffs might be used in a "stacked" configuration to effectively treat saccular abdominal aortic aneurysm (AAA). METHODS: In a recent 1-year period, 147 patients underwent endovascular AAA repair. Of these, 5 (4 men; mean age 61.6 years, range 55-69) had saccular AAAs with a mean diameter of 3.7 +/- 0.2 cm (range 3.0-4.7). AneuRx aortic cuff prostheses (3.75-cm length) were deployed sequentially in these 5 patients via a right femoral approach; the devices were overlapped approximately 1.5 to 2.0 cm until complete exclusion of the aneurysm was achieved. Endograft surveillance was performed using computed tomography at 4 weeks postoperatively and then every 6 months. RESULTS: Successful exclusion of the saccular infrarenal aortic aneurysms was achieved in all 5 patients using 2 or 3 "stacked" stent-graft extensions. Four of the 5 procedures were performed under spinal anesthesia; the average procedural time was 96 +/- 41 minutes. The average hospital length of stay was 1.6 days; no major morbidity or mortality was encountered. There were no early or late endoleaks, aneurysm expansion, or device migration over a follow-up that ranged to 12 months. CONCLUSIONS: Saccular AAA provides ideal anatomy for endovascular repair with a "tube" endograft. "Stacked" aortic cuffs create a customized stent-graft that is not otherwise commercially available. PMID- 12096944 TI - Single-center experience with carotid stent restenosis. AB - PURPOSE: To report the angiographic morphology of carotid stent restenosis and the possible therapies based on data from a single-center experience. METHODS: In a 45-month period, 279 patients (196 men; mean age 70 +/- 9 years, range 50-89) underwent successful Wallstent placement in 303 stenotic internal carotid arteries (ICA). Patients were followed with duplex sonography; angiography was used to confirm any significant (> or =70%) recurrent lesions detected on the ultrasound scan. Further balloon dilation with or without stent placement was undertaken. RESULTS; Over a median 12-month follow-up (interquartile range 6-24), there were 9 (3.0%) carotid stent restenoses found, all within 12 months after stent placement. Two types of restenosis were differentiated. In the more common form, "in-stent" stenoses (n = 6) were detected and treated with stent placement; lasting success (patency >12 months after retreatment) was achieved in 4. Early second and third recurrences arose in the other 2 stents within 3 months of the first retreatment; additional stents were placed at each recurrence. Both patients suffered a major cerebral event after 17 months. Less often, an "end of stent" stenosis (n = 3) developed at a kink in the ICA adjacent to the cephalad end of the stent. Lasting success was achieved by balloon dilation without additional stent placement in all 3 cases. No procedure-related complications were observed within 30 days after any treatment for restenosis. CONCLUSIONS: Carotid stent restenosis, which occurs rarely after 1 year, can be treated safely by further percutaneous interventions. PMID- 12096945 TI - Coil embolization combined with carotid-subclavian bypass for treatment of subclavian artery aneurysm. AB - PURPOSE: To report 2 cases of proximal subclavian artery aneurysm treated with carotid-subclavian bypass and coil embolization. CASE REPORTS: A subclavian artery aneurysm was discovered incidentally during presurgical coronary angiography in 2 men (72 and 63 years of age). Both patients underwent planned carotid endarterectomy, during which a carotid-subclavian bypass was created with a Dacron graft; the distal subclavian artery was also ligated. The aneurysm was completely excluded from the circulation by coil embolization in a separate procedure. The patients were discharged after 3 days with no adverse events. Magnetic resonance angiography at 6 months showed continued aneurysm exclusion in both cases. The patients continue to be well >1 year after treatment. CONCLUSIONS: Embolization with supraclavicular bypass represents a viable alternative to traditional thoracotomy and subclavian artery reconstruction for treatment of subclavian artery aneurysms. PMID- 12096946 TI - Endoventricular transplantation of allogenic skeletal myoblasts in a porcine model of myocardial infarction. AB - PURPOSE: To assess the technical feasibility of percutaneous endoventricular injection of skeletal myoblasts into an infarcted porcine myocardium. METHODS: A skeletal muscle biopsy was obtained from a donor pig and processed for myoblast expansion in vitro. Myocardial infarction was induced in a host pig via fibrin coil placement in the left anterior descending artery. Four weeks later, electromechanical mapping of the left ventricle identified the infarction site, into which approximately 200 million allogenic cells obtained from the muscle biopsy were directly injected (0.1 mL/injection at 25 sites) from inside the ventricular cavity via a needle injection catheter inserted through the femoral artery. Ten days after transplantation, the injected heart was evaluated histologically for the presence of myoblasts. RESULTS: Electrocardiography, echocardiography, left ventricular angiography, and electromechanical mapping confirmed the myocardial infarction. During the cell transfer procedure, premature ventricular contractions confirmed needle placement in the endocardium. Histological evaluation of the host heart 10 days after cell transfer revealed living myoblasts and multinucleated myotubes in the infarcted region, indicating successful transplantation. CONCLUSIONS: Direct myoblast transplantation into infarcted porcine myocardium using an endoventricular injection was technically feasible. The results in this model show that transplanted myoblasts were able to survive for 10 days after transplantation. PMID- 12096947 TI - Histological changes in canine aorta 1 year after stent-graft implantation: implications for the long-term stability of device anchoring zones. AB - PURPOSE: To examine aortic histology 1 year after stent-graft implantation in a canine model as a means of assessing the durability of endograft fixation. METHODS: Fourteen mongrel dogs each received 1 stent-graft and 1 bare stent placed endoluminally in the abdominal aorta. Eight animals were followed for 1 year, 3 for 4 to 8 weeks, and the remaining 3 for 24 to 48 hours. Aortic specimens were stained with hematoxylin-eosin, elastica-van Gieson, and Masson's trichrome and examined with light and electron microscopy and immunohistochemistry to identify smooth muscle cells (SMC), endothelialization, aortic wall ultrastructure, and changes at the device anchoring sites. RESULTS: No dilatation or dissection was noted at any of the device anchoring sites. The aortic media at 1 year was remarkably decreased in thickness: 891 +/- 196 microm in the control tissue versus 388 +/- 70 microm for the proximal stent-grafted aorta and 457 +/- 148 microm for the bare-stented aortic segment. Other important histological features were reduced elastic lamellae in the stent-grafted aorta versus control (p<0.0001), increased SMC density in the stent-grafted aortic region (p<0.0001 versus control), and absence of inflammatory infiltrate. Complete neointimal covering and endothelialization of the luminal endograft surface were found. SMCs generally showed no ultrastructural features of necrosis. CONCLUSIONS: Aortic stent-grafts induce distinctive histological changes in the aortic wall at 1 year, even when implanted in a healthy aorta. Although there is considerable medial elastin loss, an increased medial SMC density, an exuberant neointima, and a general absence of perigraft inflammation suggest an ongoing process of structural restoration at the device anchoring sites. PMID- 12096948 TI - Abdominal aortic healing associated with a thin-walled Dacron-covered endovascular graft in a canine model. AB - PURPOSE: To characterize the healing response associated with an experimental endovascular graft (EVG) by examining the lumen of the prosthesis and the native vessel wall responses after implantation in a canine model. METHODS: An endovascular graft (EVG) constructed of hookless self-expanding nitinol springs covered by an ultrathin Dacron fabric was placed in the abdominal aorta of 8 dogs following femoral artery catheterization. After 12 weeks, specimens were subjected to histological and immunocytochemical testing to quantitatively and qualitatively analyze the cellular makeup of the luminal and abluminal tissues. RESULTS: Gross examination revealed a glistening, thrombus-free luminal surface on all prostheses, which was confirmed by scanning electron microscopy. The EVGs were well incorporated into the aortic wall, leaving a concentric, smooth flow surface. Immunocytochemistry verified the presence of von Willebrand factor positive endothelial cells on the luminal surface and alpha-smooth muscle cell actin-positive smooth muscle and/or fibroblast cells in the subendothelial space. A sparse inflammatory response and a paucity of proliferating nuclear cell antigen-positive cells were noted within the neomedial lining adjacent to the graft material. In addition, a rich vaso vasorum-like plexus of microvessels was evident within the neomedial tissue. The native vessel wall upon which the EVG impinged was largely unaffected. CONCLUSIONS; These data demonstrate that placement of an experimental EVG in the canine abdominal aorta elicits a healing response that is distinct from interpositional vascular grafts. Results from this study have particular relevance to the proximal and distal anastomotic neck regions. PMID- 12096949 TI - A laparoscopic approach to the abdominal aorta for thoracic stent-graft deployment: evaluation in a porcine model. AB - PURPOSE: To develop laparoscopic techniques for aortic stent-graft placement as an alternative to the femoral approach. METHODS: Endovascular stent-grafts were placed in 8 pigs via a totally laparoscopic retroperitoneal approach. After needle puncture, a guidewire was inserted into the abdominal aorta, followed by an 18-F sheath through which a Talent stent-graft was deployed in the descending thoracic aorta without aortic clamping. All the endovascular tools were inserted into the retroperitoneal area via the ports. After the sheath was withdrawn, hemostasis was achieved by suturing the aortic puncture under aortic cross clamping. After sacrificing the animals, the thoracic aorta was removed to verify the position and deployment of the stent-grafts. RESULTS: Seven (87.5%) of 8 procedures were successfully completed; the first animal died from hemorrhage due to inadvertent injury to the posterior infrarenal aortic wall. The accurate deployment and position of the stent-grafts were verified visually after sacrifice. Mean (+/- SD) procedural, implantation, and aortic cross-clamping times were 205 +/- 56, 22 +/- 9, and 30 +/- 19 minutes, respectively. Mean blood loss was 120 +/- 56 mL. CONCLUSIONS: Thoracic aortic stent-grafting using a laparoscopic approach to the infrarenal aorta is feasible. More studies will be required to define the place of combined endovascular and laparoscopic procedures as an alternative to the femoral surgical approach for stent-graft placement. PMID- 12096950 TI - Endovascular brachytherapy for prevention of recurrent renal in-stent restenosis. AB - PURPOSE: To examine the ability of endovascular brachytherapy to prevent recurrent in-stent restenosis in patients at high risk for this complication. METHODS: Thirteen renal stent patients (8 women; mean age 66 +/- 8 years) with an initial (n=9) or recurrent (n=4) in-stent restenosis underwent redilation followed by high-dose-rate brachytherapy (12 Gy of gamma radiation delivered to the target site 5 mm from an iridium-192 source axis). RESULTS: The procedure was technically successful in 11 (85%) patients; in the other 2, the renal artery could not be accessed with the large sheaths required for brachytherapy. One patient with no clinical suspicion of restenosis died of an unrelated cause during the 1-year follow-up. Eight (80%) of 10 patients alive at 1 year had no in stent restenosis apparent on duplex sonography or angiography. One of the postradiation recurrent restenoses was redilated, but the other patient was treated conservatively. CONCLUSIONS; Renal angioplasty followed by brachytherapy seems to be a feasible and efficient method to prevent recurrent in-stent restenosis in renal arteries at increased risk. PMID- 12096951 TI - Endovascular management of blunt traumatic renal artery dissection. AB - PURPOSE: To describe the importance of accurate diagnosis and successful emergent endovascular repair of intimal injury to the renal artery from sudden deceleration. CASE REPORT: A 22-year-old man was evaluated after jumping head first out of a fourth floor window. Gross hematuria was detected at the initial assessment. Contrast-enhanced computed tomography (CT) revealed diminished uptake in the right kidney. Arteriographic examination of the renal artery demonstrated 2 areas of intimal disruption, which a team of physicians decided to treat with angioplasty and stent placement owing to the associated intracranial injuries. Two balloon-expandable stents were successfully deployed to exclude the intimal defects. Once the associated injuries were also addressed, the patient made an uneventful recovery. CONCLUSIONS; Management of blunt traumatic injuries to the renal vasculature involves timely assessment to maximize preservation of renal function. In the setting of multiple concomitant injuries, recent experience supports the consideration of endoluminal intervention for traumatic renal artery dissections. PMID- 12096952 TI - Endoluminal stent-graft repair of a renal artery aneurysm. AB - PURPOSE: To present a renal artery aneurysm that was treated successfully by endoluminal grafting. CASE REPORT: A 48-year-old woman presented with a 2.5-cm saccular left renal artery aneurysm. A Jostent coronary stent-graft was placed transluminally into the left renal artery via a femoral artery cutdown. After 12 months, renal function remained normal, and computed tomography revealed normal kidney perfusion with complete resolution of the renal artery aneurysm. CONCLUSIONS; The advances in endovascular stent-grafts will allow more visceral artery aneurysms to be treated with a minimally invasive approach. PMID- 12096954 TI - Diameter of occluded superficial femoral arteries limits percutaneous recanalization: preliminary results. AB - PURPOSE: To explore if the diameter of an occluded superficial femoral artery (SFA) can predict the technical success of percutaneous recanalization. METHODS: Two hundred patients (151 men; mean age 57 +/- 16 years) suffering from peripheral arterial occlusive disease were evaluated with duplex ultrasonography to measure arterial diameters and percent diameter reduction. Seventy-nine (39.5%) patients had segmental or complete SFA occlusions that ranged in age from 2 to 26 months (median 11). The majority of occlusions (45, 57.0%) were < or =10 cm long (range 10-35). Percutaneous recanalization was performed in 71 patients using either conventional angioplasty or local low-dose thrombolysis with rtPA. RESULTS: Diameters of 79 unilaterally occluded SFAs at the level of occlusion were 4.5 +/- 1.4 mm versus 5.7 +/- 1.3 mm for the patent contralateral artery (p=0.055). None of the 121 patent SFAs had diameters < or =3.5 mm, but 12 (15%) of the 79 occluded SFAs did, and none of these could be recanalized. The sensitivity of the 3.5-mm cut point to predict recanalization failure was 44% (specificity 100%). CONCLUSIONS; Our data suggest that occluded SFAs with a diameter reduction to < or =3.5 mm are not suitable for percutaneous recanalization, as the original arterial lumen cannot be reconstructed. PMID- 12096953 TI - Percutaneous reconstruction of the aortoiliac bifurcation with the "kissing stents" technique: long-term follow-up in 106 patients. AB - PURPOSE: To evaluate the long-term results using the kissing stents technique for treatment of occlusive disease involving the aortoiliac bifurcation. METHODS: One hundred six patients (97 men; mean age 52.5 +/- 10.2 years, range 33-78) were treated with the kissing stents technique for bilateral aortoiliac stenosis (55, 51.9%), unilateral occlusion of the common iliac artery (CIA) with contralateral stenosis (47, 44.3%), and bilateral CIA occlusion (4, 3.8%). Clinical examination and duplex scans were performed prior to discharge and at 1, 6, and 12 months, followed by yearly examinations thereafter. RESULTS: Bilateral stent implantation was successful in all patients. No major procedure-related complications were observed. Self-expanding stents were deployed in 62 (58.5%) patients and balloon expandable devices in 44 (41.5%). Fifteen (7.1%) hematomas were observed at the 212 access sites. Mean follow-up was 30.1 +/- 11.1 months (range 12-137). Duplex imaging diagnosed significant (>50%) restenosis in 15 (14.8%) of 101 patients and reocclusion in 4 (4%); 17 (89.5%) of these patients had recurrent symptoms and all were retreated (endovascular procedure in 18 and an aortobifemoral bypass in 1). Primary and secondary cumulative patency rates at 36 months were 79.4% and 97.7%, respectively. Balloon-expandable stents had a nonsignificantly higher patency rate compared to self-expanding stents. CONCLUSIONS: Based on our experience, aortoiliac endovascular reconstruction with the kissing stents technique is a safe and effective procedure, representing an alternative to conventional surgery in selected patients. PMID- 12096955 TI - Stent-graft repair of a recurrent popliteal arteriovenous fistula. AB - PURPOSE: To describe the successful endovascular treatment of a popliteal arteriovenous fistula with a stent-graft. CASE REPORT: A 54-year-old man was referred to our hospital with a distal popliteal arteriovenous fistula following an arthroscopic meniscectomy 6 years earlier. Three surgical attempts to close the fistula were undertaken, but the fistula recurred. He now presented with symptoms of progressive venous hypertension with claudication, swelling of the leg, and ulceration at the ankle. The fistula was closed with a stent-graft deployed percutaneously. At 18 months, the patient is doing well; duplex evaluation has documented the patency of the popliteal artery and the stent graft. CONCLUSIONS: Endovascular treatment of a popliteal arteriovenous fistula is an alternative to open surgical reconstruction. In this situation, after multiple failed surgical closures and in a leg with extensive venous hypertension, it may be the best treatment option. PMID- 12096956 TI - Endovascular stent-graft adapted to the endoluminal environment: prototype of a new endoluminal approach. PMID- 12096957 TI - Microcatheter technique assists stenting of complex carotid stenoses. PMID- 12096958 TI - Re: "Modified technique for using vascular sheaths in carotid artery stent placement". PMID- 12096959 TI - Time course of changes in corneal forward shift after excimer laser photorefractive keratectomy. AB - BACKGROUND: Excimer laser refractive surgery has been reported to induce forward shift of the cornea, but its long-term sequelae remain unknown. OBJECTIVES: To prospectively investigate the time course of changes in corneal elevation after excimer laser photorefractive keratectomy (PRK). METHODS: We performed PRK on 65 eyes of 34 patients with refractive errors of -1.25 to -10.0 diopters. The anterior/posterior corneal elevation and corneal thickness were measured with a scanning-slit corneal topography system before and 1 week and 1, 3, 6, and 12 months after surgery. Twenty eyes of 10 healthy control subjects underwent similar measurements at 3-month intervals. RESULTS: The posterior corneal surface displayed a mean +/- SD forward shift of 36.6 +/- 25.3 microm 1 week after PRK, which gradually increased to 55.1 +/- 46.1 microm at 1 year. All postoperative values were significantly larger than those of healthy controls (2.4 +/- 8.9 microm; P<.001, Mann-Whitney test). The largest forward shift occurred within the first postoperative week. The progression thereafter was most pronounced from 1 to 6 months, and nearly stabilized at 6 months. The variance of postoperative data was statistically significant (P<.001, repeated-measures analysis of variance). Multiple postoperative comparisons demonstrated significant differences between measurements at 1 week and 6 months (P =.002, Tukey Honestly Significant Difference), at 1 week and 1 year (P<.001), at 1 and 6 months (P<.001), and at 1 month and 1 year (P<.001). Progression of forward shift was more prominent in eyes with less preoperative corneal thickness and greater myopia that required larger laser ablation. We observed no progressive thinning and expansion of the cornea during the 1-year follow-up, which refuted the occurrence of true ectasia. A statistically significant correlation was found between the amount of myopic regression and the forward shift of the cornea (Pearson correlation coefficient, r = -0.37; P =.005). CONCLUSIONS: Photorefreactive keratectomy induced forward shift of the cornea, which is not true corneal ectasia. The largest forward shift occurred within the first postoperative week. Changes were progressive up to 6 months postoperatively, but became almost stable thereafter. Eyes with thinner cornea and higher myopia, requiring greater photoablation, are more predisposed to progression. Forward shift of both corneal surfaces added to the tendency toward myopic regression after PRK. PMID- 12096960 TI - Measurement of the magnitude and axis of corneal polarization with scanning laser polarimetry. AB - BACKGROUND: Scanning laser polarimetry uses a polarization compensator to isolate corneal birefringence from the birefringence of the retinal nerve fiber layer. This compensator assumes a fixed corneal polarization magnitude (CPM) of 60 nm and a fixed corneal polarization axis (CPA) of 15 degrees in all subjects. OBJECTIVES: To measure the CPM and CPA with a scanning laser polarimeter and to determine if the assumed compensation values are representative of those observed in healthy and glaucomatous eyes. METHODS: The CPM and CPA were measured in 51 healthy eyes and 55 glaucomatous eyes using a modified scanning laser polarimeter (GDx Nerve Fiber Analyzer; Laser Diagnostic Technologies Inc, San Diego, Calif) with an experimental variable CPM and CPA compensator. The CPM and CPA distributions in healthy and glaucomatous eyes were compared, and the CPM and CPA relationships with age, corneal thickness, and corneal curvature were also investigated. Nasally upward CPA values (in degrees) were recorded as negative; nasally downward CPA values were recorded as positive. RESULTS: The CPM and CPA measurements were normally distributed with many eyes having values different from those assumed by the GDx corneal compensator. For healthy and glaucomatous eyes combined, CPM measurements ranged from 7 nm to 91 nm (mean +/- SD, 40.0 +/- 15.7 nm). The CPA measurements ranged from -13 degrees to 73 degrees (mean +/- SD, 24.5 degrees +/- 17.4 degrees ). A significant effect of age on CPA was observed when all eyes were combined (R(2) = 0.10; P<.001). There were no differences in CPM or CPA between healthy and glaucomatous eyes after adjusting for age. No effects of corneal thickness on CPM (R(2) = 0.04; P =.05) or CPA (R(2) = 0.01; P =.24) or of corneal curvature on CPM (R(2) = 0.002; P =.67) or CPA (R(2) = 0.009; P =.34) were observed. CONCLUSIONS: The range of CPM and CPA values observed in glaucomatous and healthy eyes suggests that the narrow-band corneal compensator used by the GDx scanning laser polarimeter is inappropriately compensating for anterior segment birefringence in many eyes. PMID- 12096961 TI - The structure-function relationship in eyes with glaucomatous visual field loss that crosses the horizontal meridian. AB - OBJECTIVE: To evaluate the relationship between visual field loss and glaucomatous optic discs in eyes in which field loss spreads across the horizontal meridian. SUBJECTS AND METHODS: Ninety-six patients with glaucoma (9 advanced, 60 moderate, and 27 early) with 2 successive abnormal fields were included. Standard achromatic automated perimetry defects were identified with a nerve fiber bundle map to identify abnormal sectors. Crossover was present if the superior and inferior sectors at the horizontal meridian (nasal, central, or temporal) were both abnormal. Optic disc damage was assessed by masked grading of simultaneous stereophotographs. RESULTS: Only 30% (29) of glaucomatous eyes showed crossover, and only 2 of those eyes had early loss. The most frequent pattern of visual field loss (41% of eyes) was single hemifield damage with defects in contiguous sectors. Regardless of the pattern or severity of visual loss, most eyes (66 [69%] of 96) had both superior and inferior optic disc damage. CONCLUSIONS: Early glaucomatous visual field loss rarely crosses the horizontal meridian, but defects in both hemifields at the horizontal meridian are more common in more advanced field loss. In 26 (90%) of 29 eyes with crossover, it could be explained by changes at the optic nerve head. PMID- 12096962 TI - Latanoprost and timolol combination therapy vs monotherapy: one-year randomized trial. AB - OBJECTIVE: To compare the efficacy and safety of a fixed combination of 0.005% latanoprost and 0.5% timolol maleate administered once daily vs monotherapy with either 0.005% latanoprost once daily or 0.5% timolol twice daily. METHODS: Patients with either primary or secondary open-angle glaucoma or ocular hypertension participated in a 6-month, randomized, double-masked, multicenter study with 3 parallel treatment groups. The double-masked period was preceded by a 2- to 4-week "run-in" treatment with timolol. Subjects could receive fixed combination therapy during a 6-month open-label extension. MAIN OUTCOME MEASURE: The difference between groups in mean diurnal intraocular pressure reduction in study eye(s) from baseline through 6 months of treatment. RESULTS: Overall, 418 patients were enrolled in the study; 332 completed the open-label phase. Diurnal intraocular pressure levels were similar at baseline, but at week 26, they were 19.9 +/- 3.4 mm Hg in the fixed combination therapy group, 20.8 +/- 4.6 mm Hg in latanoprost-treated patients, and 23.4 +/- 5.4 mm Hg in timolol-treated patients (data are given as mean +/- SD). The mean change from baseline was greater among patients receiving fixed combination therapy compared with each monotherapy group (P<.01). Fixed combination therapy effectively lowered intraocular pressure levels for up to 1 year. All treatments were well tolerated. CONCLUSION: The combination of 0.005% latanoprost and 0.5% timolol administered once daily is effective and well tolerated for up to 12 months. PMID- 12096963 TI - Postenucleation adjuvant therapy in high-risk retinoblastoma. AB - PURPOSE: The main purpose of this study was to determine the efficacy of postenucleation adjuvant therapy in preventing metastasis in cases of high-risk retinoblastoma. METHODS: This was a retrospective, nonrandomized comparative study. Of 1020 consecutive patients with retinoblastoma had were managed at a referral center between January 1974 and December 1999, 80 (8%) of those analyzed had unilateral sporadic cases that were treated by primary enucleation and that had high-risk characteristics for metastasis on histopathology reports (anterior chamber seeding, iris infiltration, ciliary body infiltration, massive choroidal infiltration, invasion of optic nerve lamina cribrosa, retrolaminar optic nerve invasion, invasion of optic nerve transection, scleral infiltration, and extrascleral extension). The main outcome measure was the development of metastasis at a minimum follow-up period of 12 months. RESULTS: There were 44 male and 36 female patients, with age ranging from 1 day to 16 years (median, 33 months). A single histopathologic high-risk characteristic was present in 50 patients (62.5%). Thirty patients (37.5%) manifested 2 or more high-risk characteristics. Forty-six patients (58%) had received postenucleation adjuvant therapy (chemotherapy with or without orbital external beam radiotherapy). Adjuvant therapy was not administered in 34 patients (42%). Metastasis occurred in 10 patients (13%) at a median of 9 months (range, 6-57 months) following enucleation. Eight (80%) of those who developed metastasis had not received adjuvant therapy. A significant difference (P =.02) was found in the incidence of metastasis between the group that had received adjuvant therapy (4%; 2/46) and the group that had not (24%; 8/34). The beneficial effect of adjuvant therapy was statistically significant in subgroups of patients with massive choroidal infiltration (P =.04) or retrolaminar optic nerve invasion (P =.04). There were no adjuvant therapy-related serious systemic complications. CONCLUSION: Postenucleation adjuvant therapy is safe and effective in significantly reducing the occurrence of metastasis in patients with retinoblastoma manifesting histopathologic high-risk characteristics. PMID- 12096964 TI - Combined plaque radiotherapy and transpupillary thermotherapy for choroidal melanoma: tumor control and treatment complications in 270 consecutive patients. AB - OBJECTIVE: To evaluate tumor control and treatment complications following plaque radiotherapy combined with transpupillary thermotherapy for choroidal melanoma. DESIGN: Prospective noncomparative interventional case series. INTERVENTION: All patients received treatment for choroidal melanoma using plaque radiotherapy followed by 3 sessions of transpupillary thermotherapy provided at plaque removal and at 4-month intervals. PARTICIPANTS: Two hundred seventy patients with newly diagnosed choroidal melanoma. MAIN OUTCOME MEASURES: The 2 main outcome measures included local tumor recurrence and treatment-related complications. The clinical data regarding patient features, tumor features, radiotherapy and thermotherapy parameters were analyzed for their effect on the 2 main outcomes using Cox proportional hazards regression models. RESULTS: Prior to treatment, the median base of the tumor was 11 mm (range, 4-21 mm) and the median thickness was 4 mm (range, 2-9 mm). Most tumors were located in the posterior pole with a median proximity of 2 mm to the foveola and 2 mm to the optic disc. The median radiotherapy dose to the tumor apex was 9000 rad. Transpupillary thermotherapy was applied in 3 sessions at 4-month intervals for a median of 700 mW. The tumor decreased in thickness to a median of 2.3 mm by 1 year and 2.1 mm by 2 years' follow-up with stable findings thereafter. Using Kaplan-Meier estimates, tumor recurrence was 2% at 2 years and 3% at 5 years. Risk factors for tumor recurrence included macular location of the tumor epicenter (P =.03), diffuse tumor configuration (P =.005), and tumor margin extending underneath the foveola (P =.001). Using Kaplan-Meier estimates, treatment-related complications at 5 years included maculopathy in 18% of the participants, papillopathy in 38%, macular retinal vascular obstruction in 18%, vitreous hemorrhage in 18%, rhegmatogenous retinal detachment in 2%, cataract in 6%, and neovascular glaucoma in 7%. Enucleation for radiation complications was necessary in 3 cases (1%). CONCLUSION: Plaque radiotherapy combined with transpupillary thermotherapy provides excellent local tumor control with only 3% recurrence at 5 years' follow up. PMID- 12096965 TI - Adenovirus-mediated gene therapy using human p21WAF-1/Cip-1 to prevent wound healing in a rabbit model of glaucoma filtration surgery. AB - OBJECTIVE: To determine if adenovirus-mediated p21(WAF-1/Cip-1) (p21) gene therapy can prevent fibroproliferation and wound healing in a rabbit model of glaucoma filtration surgery. METHODS: In vitro studies were performed using rabbit Tenon fibroblasts harvested from fresh tissue. In vivo studies were conducted in New Zealand white rabbits. A full-thickness sclerotomy was performed under a limbal-based conjunctival flap. Reagents tested included a replication deficient recombinant adenovirus containing the human p21 gene (rAd.p21); the nonspecific marker gene for green fluorescent protein or beta-galactosidase; mitomycin, 0.5 mg/mL; and balanced saline solution. Each treatment was applied episclerally for 5 minutes before the sclerotomy using a soaked cellulose sponge placed under the surgically created conjunctival flap. Independent experiments were conducted to (1) monitor changes in intraocular pressure during a 30-day period after treatment and examine surgical site histological features, (2) examine changes in bleb morphologic features over 30 days, (3) determine outflow facility 14 days after treatment, and (4) examine the localization and persistence of rAd.p21 expression between 3 and 60 days after treatment. RESULTS: Treatment of Tenon fibroblasts with rAd.p21 resulted in a dose-dependent inhibition of DNA synthesis and cell growth in vitro. In vivo, rAd.p21 inhibited wound healing and fibroproliferation after filtration surgery, comparably to mitomycin. Mitomycin caused notable thinning of the bleb wall. In addition, 2 of the 5 mitomycin-treated eyes exhibited an abscess with hypopyon and hyalitis 30 days after surgery, which was not observed in any of the rAd.p21-treated eyes. None of the treatments resulted in a significantly sustained decrease in intraocular pressure during the 30-day period, although mitomycin treatment resulted in a significant (P =.02) increase in outflow facility 2 weeks after surgery in separate animals. Mitomycin- and rAd.p21-treated eyes had functioning blebs at the end of the experiment based on slitlamp examination. CONCLUSIONS: Mitomycin and rAd.p21 were effective in preventing fibroproliferation and wound healing in a rabbit model of glaucoma surgery. Mitomycin treatment increased outflow facility in normal-pressure eyes. CLINICAL RELEVANCE: Gene therapy with rAd.p21 may provide an effective antiproliferative for glaucoma filtration surgery, without the complications associated with mitomycin. PMID- 12096966 TI - Effects of topical anti-inflammatory and antiallergic eyedrops on prostaglandin E2-induced aqueous flare elevation in pigmented rabbits. AB - OBJECTIVE: To evaluate the role of topical instillation of anti-inflammatory or antiallergic agents on experimental elevation of aqueous flare induced by prostaglandin E2 (PGE2) in pigmented rabbits. METHODS: Transcorneal diffusion of PGE2, 25 microg/mL (7.09 x 10 (-2)mmol/L), by means of a glass cylinder produced aqueous flare elevation. Anti-inflammatory or antiallergic agents were topically administered once or twice before PGE2 application. Aqueous flare was measured with a laser flare-cell meter. Results are given as mean +/- SD. RESULTS: Double instillations of 0.1% betamethasone sodium phosphate and 0.1% fluorometholone acetate at 4 and 2 hours before PGE2 application inhibited 61% +/- 11% and 46% +/ 14%, respectively, of flare elevation. Double instillations of 0.1% diclofenac sodium and 0.1% pranoprofen at 4 and 2 hours before PGE2 application did not inhibit flare elevation. Double instillations of 0.1% betamethasone, 0.1% fluorometholone, 0.1% diclofenac, and 0.1% pranoprofen at 1 and 0.5 hour before PGE(2) application inhibited 16% +/- 10%, 16% +/- 6%, 24% +/- 9%, and 23% +/- 10%, respectively, of flare elevation. Double instillations of 2% cromolyn sodium, 0.5% tranilast, 0.025% levocabastine hydrochloride, 0.1% pemirolast potassium, and 0.01% ibudilast at 1 and 0.5 hour before PGE2 application did not inhibit flare elevation. Single instillation of 0.1% betamethasone 6 hours before PGE2 application inhibited 88% of PGE2-induced aqueous flare elevation. Single instillation of 0.1% diclofenac 1 hour before PGE2 application inhibited 23% of PGE2-induced aqueous flare elevation. CONCLUSIONS: Betamethasone needed several hours after topical instillation to inhibit flare elevation, but diclofenac needed 1 hour. Antiallergic agents did not affect disruption of the blood-aqueous barrier in rabbits. CLINICAL RELEVANCE: Corticosteroid eyedrops may need several hours from instillation to show action. PMID- 12096967 TI - Incident open-angle glaucoma and blood pressure. AB - BACKGROUND: The risk of open-angle glaucoma (OAG) may be related to low blood pressure (BP) relative to intraocular pressure (IOP), ie, to low perfusion pressure (PP). Alternatively, systemic hypertension may increase OAG risk. OBJECTIVE: To clarify these possible relationships by evaluating hypertension and PP (where PP = BP - IOP) as risk factors for incident OAG in a black population. DESIGN: Population-based cohort study (85% participation); simple random sample of residents of Barbados, West Indies, aged 40 years and older. PARTICIPANTS: Two thousand nine hundred eighty-nine black participants at risk; 67 developed OAG after 4 years (2.2% incidence). MAIN OUTCOME MEASURE: Adjusted relative risk (RR) of OAG from logistic regression analyses. RESULTS: The 4-year risk increased markedly with baseline IOP. With an IOP less than or equal to 17 mm Hg, incidence was 0.7%, increasing to 18.3% with IOP greater than 25 mm Hg, for a 25-fold increase in RR. However, OAG developed throughout the IOP range and two thirds of incident cases had baseline IOP less than 25 mm Hg. Baseline hypertension was associated with a halving of the RR of OAG (RR, 0.49; 95% confidence interval [CI], 0.29-0.85); the RR also tended to decrease as systolic BP increased (P =.07). Consistent with these findings, a lower baseline PP increased RR (systolic PP <101 mm Hg, 2.6 [95% CI, 1.3-4.9]; diastolic PP <55 mm Hg, 3.2 [95% CI, 1.6 6.6]; mean PP <42 mm Hg, 3.1 [95% CI, 1.6-6.0]). CONCLUSIONS: As baseline IOP increased, the risk of OAG substantially increased. In contrast, persons with systemic hypertension at baseline had half the RR, suggesting that hypertension does not increase (and may decrease) the 4-year risk of OAG. Lower PP at baseline increased RR approximately 3-fold, a result consistent with the vascular hypothesis of OAG pathogenesis. PMID- 12096968 TI - ELVIS: a new 24-hour culture test for detecting herpes simplex virus from ocular samples. AB - OBJECTIVE: To compare ELVIS (Enzyme Linked Virus Inducible System) (BioWhittaker, Walkersville, Md), a new, simple, 24-hour cell culture test for detecting herpes simplex virus (HSV), with standard cell culture and Herpchek (NEN, Boston, Mass) for detecting HSV in ocular specimens. METHODS: Retrospectively, 36 true-positive frozen-stock ocular samples that were cell-culture positive for HSV, and 25 true negative samples (varicella-zoster virus, adenovirus, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus viridans) were tested with ELVIS. Herpchek was processed at the time of initial clinical laboratory testing. Prospectively, 422 patients were tested for HSV with standard cell culture, ELVIS, and Herpchek. The sensitivity, specificity, positive and negative predictive values, and efficacy of ELVIS based on positive and negative cell cultures were determined. RESULTS: Retrospectively, ELVIS was 86.1% sensitive (31/36), 100% specific (25/25), and 91.8% efficient (56/61). The positive predictive value was 100% (31/31), and the negative predictive value was 83.3% (25/30). The sensitivity of ELVIS was equivalent to Herpchek (80.5%, 29/36) (P =.53). Prospectively, the sensitivity of ELVIS (84.8%, 28/33) was equivalent to that of Herpchek (84.8%, 28/33). CONCLUSIONS: ELVIS is an easy HSV diagnostic test that can provide faster positive culture results than standard cell culture, and it is equally sensitive but less time-consuming than Herpchek. PMID- 12096969 TI - A structured curriculum on ethics for ophthalmology residents is valuable. PMID- 12096970 TI - A structured curriculum on ethics for ophthalmology residents is not valuable. PMID- 12096971 TI - The big brown eyes of Samuel Pepys. AB - Samuel Pepys (1633-1703) is known for writing the finest diary in the English language. He was a man of remarkable accomplishments who transformed the English Navy, was president of the Royal Society, and was a member of the British Parliament. He survived the Great Plague and imprisonment in the Tower of London. During the years when he was writing the diary, Pepys began to experience great pain in his eyes when reading and writing and from photophobia, which caused him to give up writing the diary. Pepys also had an ultimately unjustifiable fear of blindness. PMID- 12096972 TI - Juvenile xanthogranuloma of the limbus in an adult. PMID- 12096973 TI - Bilateral sequential orbital involvement by eosinophilic granuloma. PMID- 12096974 TI - Hereditary X-linked juvenile retinoschisis: a review of the role of Muller cells. PMID- 12096975 TI - Presumed iris hemangioma associated with multiple central nervous system cavernous hemangiomas. PMID- 12096977 TI - Central retinal vein occlusion in bird-shot retinochoroidopathy. PMID- 12096976 TI - Retrobulbar optic neuritis associated with infliximab. PMID- 12096978 TI - Necrotizing herpetic retinopathy associated with Ramsay Hunt syndrome. PMID- 12096979 TI - Chronic lymphocytic leukemia of the orbit. PMID- 12096980 TI - Acute annular outer retinopathy. PMID- 12096981 TI - Retinopathy as a result of long-term use of thioridazine. PMID- 12096982 TI - Resolution of punctate inner choroidopathy lesions with oral prednisone therapy. PMID- 12096983 TI - Age-related eye disease study caveats. PMID- 12096987 TI - Generation of macroscopic pair-correlated atomic beams by four-wave mixing in Bose-Einstein condensates. AB - By colliding two Bose-Einstein condensates, we have observed strong bosonic stimulation of the elastic scattering process. When a weak input beam was applied as a seed, it was amplified by a factor of 20. This large gain atomic four-wave mixing resulted in the generation of two macroscopically occupied pair-correlated atomic beams. PMID- 12096985 TI - An accurate comparison of bimatoprost's efficacy and adverse effects. PMID- 12096988 TI - Forward quark jets from protons shattering the color glass condensate. AB - We consider the single-inclusive minijet cross section in pA at forward rapidity within the color glass condensate model of high energy collisions. We show that the nucleus appears black to the incident quarks except for very large impact parameters. A markedly flatter p(t) distribution as compared to QCD in the dilute perturbative limit is predicted for transverse momenta about the saturation scale, which could be as large as Q(2)(s) approximately 10 GeV2 for a gold nucleus boosted to rapidity approximately 10 (as at the BNL-RHIC). PMID- 12096989 TI - Prompt proton decay scheme of (59)Cu. AB - Five prompt proton decay lines have been identified between deformed states in (59)Cu and three spherical states in (58)Ni by means of high-resolution in-beam particle-gamma gamma coincidence spectroscopy. The GAMMASPHERE array coupled to dedicated ancillary detectors including four Delta E-E silicon strip detectors was used to study high-spin states in (59)Cu. The multiple discrete proton lines are found to probe the wave functions of states in the decay-out regime of well- and superdeformed states. PMID- 12096990 TI - Laser-induced recollision phenomena: interference resonances at channel closings. AB - An analytical theory of the resonancelike phenomena in high-order above-threshold ionization is presented that explains details of the experimental spectra and theoretical simulations. It traces the observed features to the constructive interference of "quantum orbits" with long travel times at laser intensities where the N-photon ionization channels close. Characteristic differences show up between even and odd N. The effects are generic to all laser-induced recollision phenomena. For nonsequential double ionization, their signature in the momentum distribution of the final electrons is identified. PMID- 12096991 TI - Memory in time-dependent density functional theory. AB - Exact time-dependent density functionals remember both the entire history of the density and the initial wave function. We show that the two effects are intimately related, and all history dependence can be written as initial-state dependence, including that of the exchange-correlation kernel. For states that can be evolved from a ground state, all initial-state dependence is a dependence on a pseudo-prehistory, providing a route to excited-state densities from time dependent density functional theory. PMID- 12096992 TI - Measurement of the electron electric dipole moment using YbF molecules. AB - The most sensitive measurements of the electron electric dipole moment d(e) have previously been made using heavy atoms. Heavy polar molecules offer a greater sensitivity to d(e) because the interaction energy to be measured is typically 10(3) times larger than in a heavy atom. We have used YbF to make the first measurement of this kind. Together, the large interaction energy and the strong tensor polarizability of the molecule make our experiment essentially free of the systematic errors that currently limit d(e) measurements in atoms. Our first result d(e) = (-0.2+/-3.2)x10(-26)e cm is less sensitive than the best atom measurement but is limited only by counting statistics and demonstrates the power of the method. PMID- 12096993 TI - Wave properties of a methyl group under ambient conditions. AB - Liquid-phase NMR studies on hindered rotation of methyl group in a 9 methyltriptycene derivative are reported where the standard, classical jump model of the methyl dynamics proves inadequate. On the other hand, accurate reproduction of the observed NMR line shape effects is afforded by the use of a recent quantum mechanical model in which the relevant methyl dynamics are described in terms of two quantum rate (coherence-damping) processes, characterized by two different rate constants. For ambient temperatures, such a direct evidence of the quantum nature of a rate process generally believed to be classical seems to have no precedence in the literature. PMID- 12096994 TI - Collisional blockade in microscopic optical dipole traps. AB - We analyze the operating regimes of a very small optical dipole trap, loaded from a magneto-optical trap, as a function of the atom loading rate, i.e., the number of atoms per second entering the dipole trap. We show that, when the dipole trap volume is small enough, a "collisional blockade" mechanism locks the average number of trapped atoms on the value 0.5 over a large range of loading rates. We also discuss the "weak loading" and "strong loading" regimes outside the blockade range, and we demonstrate experimentally the existence of these three regimes. PMID- 12096995 TI - Symmetry-dependent multielectron excitations near the C 1s ionization threshold and distortion of the shape resonance in CO(2). AB - Satellite bands accompanying the C 1s photoline for the CO2 molecule parallel to the electric vector of the incident radiation E are found to be more intense than those for CO2 perpendicular to E in the shape resonance region. This indicates that multielectron excitations are caused in part by the interaction of the outgoing C 1s photoelectron with the valence electrons. The photoelectron-impact valence excitations couple with the C 1s single-hole ionization and distort the shape resonance significantly. We assign the broad resonance at approximately 312 eV to a distorted Sigma(u) shape resonance. PMID- 12096996 TI - Rectification and phase locking for particles on symmetric two-dimensional periodic substrates. AB - We demonstrate a rectification phenomenon for overdamped particles interacting with a 2D symmetric periodic substrate when driven with a dc and a circular ac drive. As a function of longitudinal dc amplitude, the longitudinal velocity increases in a series of quantized steps distinct from Shapiro steps with transverse rectification occurring near these transitions. The rectification phenomenon is explained using symmetry arguments and a simple model. PMID- 12096997 TI - Topological shocks in Burgers turbulence. AB - The dynamics of the multidimensional randomly forced Burgers equation is studied in the limit of vanishing viscosity. It is shown both theoretically and numerically that the shocks have a universal global structure which is determined by the topology of the configuration space. This structure is shown to be particularly rigid for the case of periodic boundary conditions. PMID- 12096999 TI - Electron heating in the measurement of electron temperature by Thomson scattering: are thermal plasmas thermal? AB - Thomson scattering measurements have yielded electron temperatures T(e) up to 7000 K greater than the ion temperature in 1 bar thermal plasmas. To account for laser heating of electrons, T(e) was measured as a function of laser pulse energy, and an unperturbed T(e) obtained by linear extrapolation to zero pulse energy. It is shown that the absorption of laser energy by the electrons, and the cooling of the electrons by collisions and radiative emission, depend strongly on T(e). Considering all these processes gives T(e) values that are in much closer agreement with the ion temperature. PMID- 12096998 TI - Fast electron transport in ultraintense laser pulse interaction with solid targets by rear-side self-radiation diagnostics. AB - We report on rear-side optical self-emission results from ultraintense laser pulse interactions with solid targets. A prompt emission associated with a narrow electron jet has been observed up to aluminum target thicknesses of 400 microm with a typical spreading half-angle of 17 degrees. The quantitative results on the emitted energy are consistent with models where the optical emission is due to transition radiation of electrons reaching the back surface of the target or due to a synchrotron-type radiation of electrons pulled back to the target. These models associated with transport simulation results give an indication of a temperature of a few hundred keV for the fast-electron population. PMID- 12097000 TI - Coupled atomistic and discrete dislocation plasticity. AB - A computational method for multiscale modeling of plasticity is presented wherein each dislocation is treated as either an atomistic or continuum entity within a single computational framework. The method divides space into atomistic and continuum regions that communicate across a coherent boundary, detects dislocations as they approach the boundary, and seamlessly converts them from one description to another. The method permits the study of problems that are too large for fully atomistic simulation while preserving accurate atomistic details where necessary, but is currently limited to a 2D implementation. A validation test is performed by comparing the method against full atomistic simulations for a 2D nanoindentation problem. PMID- 12097001 TI - Mapping of electrostatic potential in deep submicron CMOS devices by electron holography. AB - Quantitative two-dimensional maps of electrostatic potential in device structures are obtained using off-axis electron holography with a spatial resolution of 6 nm and a sensitivity of 0.17 V. Estimates of junction depth and variation in electrostatic potential obtained by electron holography, process simulation, and secondary ion mass spectroscopy show close agreement. Measurement artifacts due to sample charging and surface "dead layers" do not need to be considered provided that proper care is taken with sample preparation. The results demonstrate that electron holography could become an effective method for quantitative 2D analysis of dopant diffusion in deep-submicron devices. PMID- 12097002 TI - Local thermal expansion in a cuprite structure: the case of Ag(2)O. AB - The local thermal behavior of the Ag(2)O framework structure has been studied by extended x-ray absorption fine structure. The average Ag-O nearest-neighbor distance expands upon heating, while the Ag-Ag next-nearest-neighbor distance contracts. An original implementation of the cumulant analysis shows that the Ag O expansion is a joint effect of potential anharmonicity and geometrical deformation of the Ag(4)O basic tetrahedral units. Accordingly, the negative thermal expansion of the lattice parameter in Ag(2)O cannot be explained uniquely in terms of rigid unit modes. PMID- 12097003 TI - Anisotropic local stress and particle hopping in a deeply supercooled liquid. AB - The origin of the microscopic motions that lead to stress relaxation in deeply supercooled liquid remains unclear. We show that in such a liquid the stress relaxation is locally anisotropic which can serve as the driving force for the hopping of the system on its free energy surface. However, not all hoppings are equally effective in relaxing the local stress, suggesting that diffusion can decouple from viscosity even at the local level. On the other hand, orientational relaxation is found to be always coupled to stress relaxation. PMID- 12097004 TI - Modulation of the nucleation rate preexponential in a low-temperature Ising system. AB - A metastable lattice gas with nearest-neighbor interactions and continuous-time dynamics is studied using a generalized Becker-Doring approach in the multidimensional space of cluster configurations. The preexponential of the metastable-state lifetime (inverse of nucleation rate) is found to exhibit distinct peaks at integer values of the inverse supersaturation. Peaks are unobservable (infinitely narrow) in the strict limit T-->0, but become detectable and eventually dominate at higher temperatures. PMID- 12097005 TI - Pattern formation in phase-separating gels with spontaneous shear. AB - We study pattern formation in gels undergoing simultaneous phase separation and orientational ordering. A 2D numerical simulation is performed using a minimal model of nonlinear elasticity with density-anisotropy coupling. For strong positive coupling, the collapsed phase elongates along the phase boundary and buckle, creating a folded structure with paired topological defects. For negative coupling, soft elasticity of the swollen phase causes a droplet morphology as in liquid-liquid phase separation. Their possible realizations in nematic liquid crystalline gels are discussed. PMID- 12097006 TI - Boundary between long-range and short-range critical behavior in systems with algebraic interactions. AB - We investigate phase transitions of two-dimensional Ising models with power-law interactions, using an efficient Monte Carlo algorithm. For slow decay, the transition is of the mean-field type; for fast decay, it belongs to the short range Ising universality class. We focus on the intermediate range, where the critical exponents depend continuously on the power law. We find that the boundary with short-range critical behavior occurs for interactions depending on distance r as r(-15/4). This answers a long-standing controversy between mutually conflicting renormalization-group analyses. PMID- 12097007 TI - Origin of the high-frequency contributions to the dielectric loss in supercooled liquids. AB - Dielectric loss spectra of four different glass formers revealing either beta relaxation or excess wing are presented. We compare the effect of pressure and temperature on the mentioned processes. Our findings suggest that the excess wing and the beta relaxation cannot be treated on the same footing. PMID- 12097008 TI - Three-dimensional single molecule rotational diffusion in glassy state polymer films. AB - Measuring three-dimensional orientational motions of many individual molecules within glassy state poly(methyl methacrylate) has enabled nanoscopic probing of bulk-obscured polymer dynamics. Complementing bulk studies, the measured distributions of nanoscale barriers to rotational motion afforded by our single molecule orientational methods directly probe the spatial heterogeneity and nanoscopic alpha-relaxation dynamics deep within the glassy state. PMID- 12097009 TI - Anomalous roughening of Hele-Shaw flows with quenched disorder. AB - The kinetic roughening of a stable oil-air interface moving in a Hele-Shaw cell that contains a quenched columnar disorder (tracks) has been studied. A capillary effect is responsible for the dynamic evolution of the resulting rough interface, which exhibits anomalous scaling. The three independent exponents needed to characterize the anomalous scaling are determined experimentally. The anomalous scaling is explained in terms of the initial acceleration and subsequent deceleration of the interface tips in the tracks coupled by mass conservation. A phenomenological model that reproduces the measured global and local exponents is introduced. PMID- 12097010 TI - Magnetization in the ultraquantum limit. AB - The magnetization below and far above the quantum limit for small Fermi surface orbits has been measured in the metallic compound LaRhIn(5). The magnetization due to a pocket of Fermi surface that comprises less than 1 part in 10(4) of the total Brillouin zone volume, and for which the quantum limit is approximately 7 T, leads to the appearance of an overall sample magnetic moment at fields between 7 and 32 T. This moment arises from diamagnetic currents produced by electrons in the ultraquantum limit. A model calculation of the origin and magnitude of the effect is in excellent agreement with the measured field dependence of the induced magnetization. PMID- 12097011 TI - Phase glass is a Bose metal: a new conducting state in two dimensions. AB - In the quantum rotor model with random exchange interactions having a nonzero mean, three phases, a (i) phase (Bose) glass, (ii) superfluid, and (iii) Mott insulator, meet at a bicritical point. We demonstrate that proximity to the bicritical point and the coupling between the energy landscape and the dissipative degrees of freedom of the phase glass lead to a metallic state at T = 0. Consequently, the phase glass is unique in that it represents a concrete example of a metallic state that is mediated by disorder, even in 2D. We propose that the experimentally observed metallic phase which intervenes between the insulator and the superconductor in a wide range of thin films is in actuality a phase glass. PMID- 12097012 TI - Why an ac magnetic field shifts the irreversibility line in type-II superconductors. AB - We show that for a thin superconducting strip placed in a transverse dc magnetic field--the typical geometry of experiments with high-T(c) superconductors--the application of a weak ac magnetic field perpendicular to the dc field generates a dc voltage in the strip. This voltage leads to the decay of the critical currents circulating in the strip, and eventually the equilibrium state of the superconductor is established. This relaxation is not due to thermally activated flux creep but to the "walking" motion of vortices in the two-dimensional critical state of the strip with in-plane ac field. Our theory explains the shaking effect that was used for detecting phase transitions of the vortex lattice in superconductors. PMID- 12097013 TI - Spin-wave quantization in ferromagnetic nickel nanowires. AB - The dynamical properties of uniform two-dimensional arrays of nickel nanowires have been investigated by inelastic light scattering. Multiple spin waves are observed that are in accordance with dipole-exchange theory predictions for the quantization of bulk spin waves. This first study of the spin-wave dynamics in ferromagnetic nanowire arrays reveals strong mode quantization effects and indications of a subtle magnetic interplay between nanowires. The results show that it is important to take proper account of these effects for the fundamental physics and future technological developments of magnetic nanowires. PMID- 12097014 TI - Reentrant spin glass behavior in Cr-doped perovskite manganite. AB - The magnetic and transport properties of the Cr-doped manganites La(0.46)Sr(0.54)Mn(1-y)Cr(y)O3 ( 0 < or = y < or = 0.08) with the A-type antiferromagnetic structure were investigated. Upon cooling, we find multiple magnetic phase transitions, i.e., paramagnetic, ferromagnetic (FM), antiferromagnetic (AFM), and spin glass in the y = 0.02 sample. The low temperature spin glass state is not a conventional spin glass with randomly oriented magnetic moments but has a reentrant character. The reentrant spin glass behavior accompanied by the anomalous multiple magnetic transitions might be due to the competing interactions between the FM phase and the A-type AFM matrix induced by the random Cr impurity. PMID- 12097015 TI - Excitation of a breather mode of bound soliton pairs in trans-polyacetylene by sub-five-femtosecond optical pulses. AB - Trans-polyacetylene with a degenerate ground state has a nonlinear excitation of soliton after photoexcitation, due to the electron-phonon coupling. The excess energy of an excited electron-hole pair over a soliton pair creation induces a breather oscillation characterized by collective stretching vibration of the carbon-carbon bonds. A time-frequency analysis of pump probe signal shows that instantaneous frequencies of stretching modes are modulated for approximately 50 fs after excitation and the modulation period is 44+/-3 fs consistent with the theoretical expectation, clearly verifying the breather. PMID- 12097016 TI - Giant infrared intensity of the Peierls mode at the neutral-ionic phase transition. AB - We present exact diagonalization results on a modified Peierls-Hubbard model for the neutral-ionic phase transition. The ground state potential energy surface and the infrared intensity of the Peierls mode point to a strong, nonlinear electron phonon coupling, with effects that are dominated by the proximity to the electronic instability rather than by electronic correlations. The huge infrared intensity of the Peierls mode at the ferroelectric transition is related to the temperature dependence of the dielectric constant of mixed-stack organic crystals. PMID- 12097017 TI - Entanglement cost of bipartite mixed states. AB - We compute the entanglement cost of several families of bipartite mixed states, including arbitrary mixtures of two Bell states. This is achieved by developing a technique that allows us to ascertain the additivity of the entanglement of formation for any state supported on specific subspaces. As a side result, the proof of the irreversibility in asymptotic local manipulations of entanglement is extended to two-qubit systems. PMID- 12097018 TI - Dynamic force spectroscopy to probe adhesion strength of living cells. AB - We studied the mechanical strength of the adhesion of living cells to model membranes. The latter contained a RGD lipopeptide which is a high affinity binding site for a cell adhesion molecule (integrin alpha(V)beta(3)). Cells adhered specifically to the vesicles. We used micropipette aspiration for breaking this adhesion with well defined forces. Systematic variation of the rate of force application revealed pronounced kinetic effects. The dependence of the detachment forces on the loading rate was well described by a power law (exponent approximately 0.4), in agreement with recent theoretical work. PMID- 12097019 TI - Optimal design, robustness, and risk aversion. AB - Highly optimized tolerance is a model of optimization in engineered systems, which gives rise to power-law distributions of failure events in such systems. The archetypal example is the highly optimized forest fire model. Here we give an analytic solution for this model which explains the origin of the power laws. We also generalize the model to incorporate risk aversion, which results in truncation of the tails of the power law so that the probability of catastrophically large events is dramatically lowered, giving the system more robustness. PMID- 12097020 TI - Global climate models violate scaling of the observed atmospheric variability. AB - We test the scaling performance of seven leading global climate models by using detrended fluctuation analysis. We analyze temperature records of six representative sites around the globe simulated by the models, for two different scenarios: (i) with greenhouse gas forcing only and (ii) with greenhouse gas plus aerosol forcing. We find that the simulated records for both scenarios fail to reproduce the universal scaling behavior of the observed records and display wide performance differences. The deviations from the scaling behavior are more pronounced in the first scenario, where also the trends are clearly overestimated. PMID- 12097021 TI - Optimal combinations of imperfect objects. AB - We consider how to make best use of imperfect objects, such as defective analog and digital components. We show that perfect, or near-perfect, devices can be constructed by taking combinations of such defects. Any remaining objects can be recycled efficiently. In addition to its practical applications, our "defect combination problem" provides a novel generalization of classical optimization problems. PMID- 12097022 TI - Comment on "Correlation induced paramagnetic ground state in FeAl". PMID- 12097023 TI - Microscopic dynamics in a strongly interacting Bose-Einstein condensate. AB - An initially stable 85Rb Bose-Einstein condensate (BEC) was subjected to a carefully controlled magnetic field pulse near a Feshbach resonance. This pulse probed the strongly interacting regime for the BEC, with the diluteness parameter (na(3)) ranging from 0.01 to 0.5. Condensate number loss resulted from the pulse, and for triangular pulses shorter than 1 ms, decreasing the pulse length actually increased the loss, until very short time scales (approximately 10 micros) were reached. The observed time dependence is very different from that expected in traditional inelastic loss processes, suggesting the presence of new microscopic BEC physics. PMID- 12097024 TI - Statistical theory of asteroid escape rates. AB - Transition states in phase space are identified and shown to regulate the rate of escape of asteroids temporarily captured in circumplanetary orbits. The transition states, similar to those occurring in chemical reaction dynamics, are then used to develop a statistical semianalytical theory for the rate of escape of asteroids temporarily captured by Mars. Theory and numerical simulations are found to agree to better than 1%. These calculations suggest that further development of transition state theory in celestial mechanics, as an alternative to large-scale numerical simulations, will be a fruitful approach to mass transport calculations. PMID- 12097027 TI - Limit on the detectability of the energy scale of inflation. AB - We show that the polarization of the cosmic microwave background can be used to detect gravity waves from inflation if the energy scale of inflation is above 2x10(15) GeV. These gravity waves generate polarization patterns with a curl, whereas (to first order in perturbation theory) density perturbations do not. The limiting "noise" arises from the second-order generation of curl from density perturbations, or rather residuals from its subtraction. We calculate optimal sky coverage and detectability limits as a function of detector sensitivity and observing time. PMID- 12097028 TI - Separation of gravitational-wave and cosmic-shear contributions to cosmic microwave background polarization. AB - Inflationary gravitational waves (GW) contribute to the curl component in the polarization of the cosmic microwave background (CMB). Cosmic shear- gravitational lensing of the CMB--converts a fraction of the dominant gradient polarization to the curl component. Higher-order correlations can be used to map the cosmic shear and subtract this contribution to the curl. Arcminute resolution will be required to pursue GW amplitudes smaller than those accessible by the Planck surveyor mission. The blurring by lensing of small-scale CMB power leads with this reconstruction technique to a minimum detectable GW amplitude corresponding to an inflation energy near 10(15) GeV. PMID- 12097029 TI - Electron mass operator in a strong magnetic field and dynamical chiral symmetry breaking. AB - The electron mass operator in a strong magnetic field is calculated. The contribution of higher Landau levels of virtual electrons, along with the ground Landau level, is shown to be essential in the leading log approximation. The effect of the electron dynamical mass generation by a magnetic field is investigated. In a model with N charged fermions, it is shown that some critical number N(cr) exists for any value of the electromagnetic coupling constant alpha, such that the fermion dynamical mass is generated with a doublet splitting for NN(cr), thus leaving the chiral symmetry unbroken. PMID- 12097025 TI - Direct evidence for neutrino flavor transformation from neutral-current interactions in the Sudbury Neutrino Observatory. AB - Observations of neutral-current nu interactions on deuterium in the Sudbury Neutrino Observatory are reported. Using the neutral current (NC), elastic scattering, and charged current reactions and assuming the standard 8B shape, the nu(e) component of the 8B solar flux is phis(e) = 1.76(+0.05)( 0.05)(stat)(+0.09)(-0.09)(syst) x 10(6) cm(-2) s(-1) for a kinetic energy threshold of 5 MeV. The non-nu(e) component is phi(mu)(tau) = 3.41(+0.45)( 0.45)(stat)(+0.48)(-0.45)(syst) x 10(6) cm(-2) s(-1), 5.3sigma greater than zero, providing strong evidence for solar nu(e) flavor transformation. The total flux measured with the NC reaction is phi(NC) = 5.09(+0.44)(-0.43)(stat)(+0.46)( 0.43)(syst) x 10(6) cm(-2) s(-1), consistent with solar models. PMID- 12097026 TI - Measurement of day and night neutrino energy spectra at SNO and constraints on neutrino mixing parameters. AB - The Sudbury Neutrino Observatory (SNO) has measured day and night solar neutrino energy spectra and rates. For charged current events, assuming an undistorted 8B spectrum, the night minus day rate is 14.0%+/-6.3%(+1.5%)(-1.4%) of the average rate. If the total flux of active neutrinos is additionally constrained to have no asymmetry, the nu(e) asymmetry is found to be 7.0%+/-4.9%(+1.3%)(-1.2%). A global solar neutrino analysis in terms of matter-enhanced oscillations of two active flavors strongly favors the large mixing angle solution. PMID- 12097030 TI - Explaining the anomalous CDF (mu)gamma missing- E(T) events with supersymmetry. AB - The CDF Collaboration recently reported an excess of events in the (mu)gamma missing E(T) (E(T)) channel that disagrees with the standard model prediction. We explain the excess via resonant smuon production with a single dominant R-parity violating coupling lambda'211, in the context of models where the gravitino is the lightest supersymmetric particle. The slepton decays to the lightest neutralino and a muon followed by neutralino decaying to a gravitino and photon. We determine a viable region of parameter space that fits the kinematical distributions of the Run I excess and illustrate the effect by examining the best fit point in detail. We provide predictions for an excess in the E(T) and photon channel at Run I and Run II. PMID- 12097033 TI - Search for nu(mu)-->nu(e) and nu(mu)-->nu(e) oscillations at NuTeV. AB - Limits on nu(mu)-->nu(e) and nu(mu)-->nu(e) oscillations are extracted using the NuTeV detector with sign-selected nu(mu) and nu(mu) beams. In nu(mu) mode, for the case of sin(2)2alpha = 1, Delta(m)(2)>2.6 eV(2) is excluded, and for Delta(m)(2)>>1000 eV(2), sin(2)2alpha>1.1 x 10(-3). The NuTeV data exclude the high Delta(m)(2) end of nu(mu)-->nu(e) oscillation parameters favored by the LSND experiment without the need to assume that the oscillation parameters for nu and nu are the same. We present the most stringent experimental limits for nu(mu)(nu(mu))-->nu(e)(nu(e)) oscillations in the large Delta(m)(2) region. PMID- 12097035 TI - Coherent vector-meson photoproduction with nuclear breakup in relativistic heavy ion collisions. AB - Relativistic heavy ions are copious sources of virtual photons. The large photon flux gives rise to a substantial photonuclear interaction probability at impact parameters where no hadronic interactions can occur. Multiple photonuclear interactions in a single collision are possible. In this Letter, we use mutual Coulomb excitation of both nuclei as a tag for moderate-impact-parameter collisions. We calculate the cross section for coherent vector-meson production accompanied by mutual excitation and show that the median impact parameter is much smaller than for untagged production. The vector-meson rapidity and transverse-momentum distribution are very different from untagged exclusive vector-meson production. PMID- 12097034 TI - Spin-dependent electron-proton scattering in the Delta-excitation region. AB - We report on measurements of the cross section and provide first data on spin correlation parameters A(TT') and A(TL') in inclusive scattering of longitudinally polarized electrons from nuclear-polarized hydrogen. Polarized electrons were injected into an electron storage ring operated at a beam energy of 720 MeV. Polarized hydrogen was produced by an atomic beam source and injected into an open-ended cylindrical cell, located in the electron storage ring. The four-momentum transfer squared ranged from Q2 = 0.2 GeV(2)/c(2) at the elastic scattering peak to Q2 = 0.11 GeV(2)/c(2) at the Delta(1232) resonance. The data provide a stringent test of pion electroproduction models. PMID- 12097036 TI - Measurement of mutual coulomb dissociation in sqrt[s(NN)] = 130 GeV Au + Au collisions. AB - We report on the first measurement of mutual Coulomb dissociation in heavy-ion collisions. We employ forward calorimeters to measure neutron multiplicity at beam rapidity. The cross section for simultaneous electromagnetic breakup of Au nuclei at a nucleon-nucleon center of mass energy sqrt[s(NN)] = 130 GeV is sigma(MCD) = 3.67+/-0.26 barns, which is comparable to the geometrical cross section. The ratio of the electromagnetic to the total cross section is in good agreement with calculations, as is the neutron multiplicity distribution. PMID- 12097037 TI - Momentum distribution of 15B fragments from the breakup of 17B. AB - The two-neutron removal cross section (sigma(-2n)) and the longitudinal momentum distribution of 15B fragments from the breakup of 17B on 9Be were measured at 70A MeV. The distribution in the projectile rest frame is characterized by a FWHM of 80+/-10 MeV/c for 15B. The sigma(-2n) is found to be 0.22+/-0.05 b. A Glauber type analysis of the data provides clear evidence of a two-neutron halo structure in 17B. PMID- 12097038 TI - Isospin mixing between low-lying states of the odd-odd N = Z nucleus 54Co. AB - Isospin mixing of the recently discovered doublet of 4(+) states with isospin quantum numbers T = 0 and T = 1 in 54Co is analyzed. It is shown that the measured E2/M1 multipole mixing ratios can be used to estimate the isospin mixing of these states. Combining the new experimental data with results of a shell model calculation, the amount of isospin mixing is found to be approximately 0.2%. PMID- 12097039 TI - Coincident fragment detection in strong field photoionization and dissociation of H2. AB - Electron-ion momentum spectroscopy is used to investigate the correlated electronic and nuclear motion in fragmentation of H2 in 4 x 10(14) W/cm(2), 25 fs laser pulses at 795 nm. Reaction channel dependent photoelectron spectra indicate that besides the main, stepwise H2 ionization H2(+) dissociation mechanism resulting in the products H(1s) + H(+) + e(-) a second new mechanism has to be assumed. The momentum distribution of H(+) ions in the dissociation channels H(1s) + H(+) + e(-) and 2H(+) + 2e(-) is found to be independent of the kinetic energy of the photoelectrons. PMID- 12097040 TI - Quantum effects in the differential cross Ssctions for the insertion reaction N(2D) + H2. AB - The quantum (QM) scattering theory has been difficult to apply to the family of insertion reactions and the approximate quasiclassical trajectory (QCT) method or statistical calculations were mostly applied. In this Letter, we compare the experimental differential cross sections for the title insertion reaction with the results of QM and QCT calculations on an ab initio potential energy surface. The QM results reproduce well the crossed beam experiment, while a small, but significant, difference in the QCT ones points to quantum effects, possibly the occurrence of tunneling through the combined potential and centrifugal barrier. PMID- 12097032 TI - Observation of chi(c2) production in B meson decay. AB - We report the first observation of chi(c2) production in B meson decays. We find an inclusive B-->chi(c2)X branching fraction of (1.80(+0.23)(-0.28)+/-0.26) x 10( 3). The data set, collected with the Belle detector at the KEKB e(+)e(-) collider, consists of 31.9 x 10(6) BB events. We also present branching fractions and momentum spectra for both chi(c1) and chi(c2) production. PMID- 12097041 TI - Influence of resonances on spectral formation of x-ray lines in Fe XVII. AB - New theoretical results from large-scale relativistic close coupling calculations reveal the precise effect of resonances in collisional excitation of x-ray lines of Ne-like Fe XVII. By employing the Breit-Pauli R-matrix method and an 89-level eigenfunction expansion, including up to n = 4 levels, significant resonance enhancement of the collision strengths of forbidden and intercombination transitions is shown. The present results should help resolve long-standing discrepancies; in particular, the present line ratios of three benchmark diagnostic lines 3C, 3D, and 3E at 15.014, 15.265, and 15.456 A, respectively, are in excellent agreement with two independent measurements on electron-beam ion traps. Strong energy dependence in cross sections due to resonances is demonstrated. PMID- 12097042 TI - Structure determination of gas-phase niobium and tantalum carbide nanocrystals via infrared spectroscopy. AB - Niobium and tantalum carbide clusters have been isolated in the gas phase and irradiated with intense tunable infrared (IR) light. Stable neutral clusters are selectively ionized and subsequently detected in a mass spectrometer. By tuning the IR frequency, infrared multiphoton absorption spectra are obtained for a whole range of clusters. These mass-selective IR spectra lead to insights into the structures of small niobium and tantalum carbide clusters and nanocrystals. PMID- 12097043 TI - Multiple filamentation of circularly polarized beams. AB - We derive a new system of equations that describes the propagation of circularly polarized laser beams in a Kerr medium. Analysis and simulations of this system show that multiple filamentation is suppressed for circularly polarized beams. PMID- 12097044 TI - Noise-enhanced phase synchronization of chaotic oscillators. AB - The effects of noise on phase synchronization (PS) of coupled chaotic oscillators are explored. In contrast to coupled periodic oscillators, noise is found to enhance phase synchronization significantly below the threshold of PS. This constructive role of noise has been verified experimentally with chaotic electrochemical oscillators of the electrodissolution of Ni in sulfuric acid solution. PMID- 12097045 TI - Large-scale finite-wavelength modulation within turbulent shear flows. AB - We show that turbulent "spirals" and "spots" observed in Taylor-Couette and plane Couette flow correspond to a turbulence-intensity modulated finite-wavelength pattern which in every respect fits the phenomenology of coupled noisy Ginzburg Landau (amplitude) equations with noise. This suggests the existence of a long wavelength instability of the homogeneous turbulence regime. PMID- 12097046 TI - New advanced operational regime on the W7-AS stellarator. AB - A promising new plasma operational regime on the Wendelstein stellarator W7-AS has been discovered. It is extant above a threshold density and characterized by flat density profiles, high energy and low impurity confinement times, and edge localized radiation. Impurity accumulation is avoided. Quasistationary discharges with line-averaged densities n(e) to 4 x 10(20) m(-3), radiation levels to 90%, and partial plasma detachment at the divertor target plates can be simultaneously realized. Energy confinement is up to twice that of a standard scaling. At B(t) = 0.9 T, an average beta value of 3.1% is achieved. The high n(e) values allow demonstration of electron Bernstein wave heating using linear mode conversion. PMID- 12097047 TI - Evidence of diffusion regions at a subsolar magnetopause crossing. AB - On 1 April 2001, the Polar satellite crossed a subsolar magnetopause associated with antiparallel magnetic fields. Over a width approximately 6 magnetosheath ion skin depths (approximately 3 magnetospheric ion skin depths), perpendicular ion flows different from E x B/B(2) as well as Hall magnetic and electric field signatures were observed. At a smaller scale, the electron flow decoupled from the magnetic field near a deep minimum in the magnetic field strength. Separatrices were identified as boundaries of low frequency electric field turbulence associated with density minima and parallel electric fields. The reconnection rate was less than 2% of the asymptotic Alfven speed. PMID- 12097049 TI - Bose-Einstein condensation at a helium surface. AB - A path integral Monte Carlo method was used to calculate the Bose-Einstein condensate fraction at the surface of a helium film at T = 0.77 K, as a function of density. Moving from the center of the slab to the surface, the condensate fraction was found to initially increase with decreasing density to a maximum value of 0.9 before decreasing. Long wavelength density correlations were observed in the static structure factor at the surface of the slab. Finally, a surface dispersion relation was calculated from imaginary-time density-density correlations. PMID- 12097048 TI - Nonlinear evolution of stimulated scatter in high-temperature plasmas. AB - Simulations of laser-plasma interactions show saturation of Raman scattering through novel subsequent Brillouin and Raman rescattering instabilities. This behavior alters the interpretation of experimental diagnostics as well as the distribution of laser energy between transmission into the target, scattering losses, and generation of energetic electrons. This paper emphasizes targets that are predicted to reach electron temperatures greater than 10 keV, a value accessible today on the Omega and Helen lasers and one which will be far higher at future facilities. In such plasmas, the nonlinear laser-plasma interaction exhibits novel features presented here. PMID- 12097050 TI - Enhanced x-ray phase determination by three-beam diffraction. AB - For decades, solving the phase problem of x-ray scattering has been a goal that, in principle, could be achieved by means of n-beam diffraction (n-BD). However, the phases extracted by the actual n-BD phasing techniques are not very precise, mainly due to systematic errors that are difficult to estimate. We present an innovative theoretical approach and experimental procedure that, combined, eliminate two major sources of error. It is a high precision phasing technique that provides the triplet-phase angle with an error of about 2 degrees. PMID- 12097051 TI - Direct role of hydrogen in the Staebler-Wronski effect in hydrogenated amorphous silicon. AB - We report a hydrogen-related defect that establishes the direct role of hydrogen in stabilizing the silicon dangling bonds created in the Staebler-Wronski effect in hydrogenated amorphous silicon. A specific NMR signal due to paired hydrogen atoms occurs only after optical excitation, exists at an intensity that is consistent with the density of optically induced silicon dangling bonds, and anneals at temperatures that are consistent with the annealing of the optically induced silicon dangling bonds. At this defect the hydrogen atoms are 2.3+/-0.2 A apart. PMID- 12097052 TI - Self-organization of a carbide superlattice during deposition of carbon on Mo. AB - Formation and evolution of a carbide superlattice (SL) during C deposition on Mo have been studied using molecular beam epitaxy techniques. The ordering of the SL is energetically driven, such that the interplay between strain and surface energies determines the length scale of the SL. Surface precipitation of C occurs within a narrow range of SL spacing that appears to control the size and spacing of the precipitates leading to a possible mechanism for nucleation of single walled carbon nanotubes. PMID- 12097054 TI - Adsorption of selenium wires in silicalite-1 zeolite: a first order transition in a microporous system. AB - A tight binding grand canonical Monte Carlo simulation of the adsorption of selenium in silicalite-1 zeolite is presented. The calculated adsorption desorption isotherms exhibit characteristic features of a first order transition, unexpected for adsorption in a microporous system with pore size of the order of 0.5 to 0.6 nm. We analyze this behavior as a result of the favored twofold coordinated chain structure of selenium that grows inside the complex three dimensional microchannel network of silicalite. This analysis is confirmed by simpler calculations of a lattice gas-type model. PMID- 12097053 TI - Laponite: aging and shear rejuvenation of a colloidal glass. AB - We study the nonlinear rheological behavior and the microscopic particle dynamics for a colloidal glass, to see whether recently developed models for driven glassy systems can be applied to predict the rheology. Qualitatively, all the findings predicted by the models can be retrieved in our system. Notably, the viscosity decreases strongly with the shear rate. Since it is difficult to predict non Newtonian viscosities of colloidal systems due to long-ranged hydrodynamic interactions, this shows the promise of this approach for predicting flow behavior. In addition, the measurements allow us to relate the microscopic diffusion dynamics to the macroscopic viscosity of the system. PMID- 12097055 TI - Adsorbate-oxide interactions during the NO + CO reaction on MgO(100) supported Pd monolayer films. AB - The potential energy diagram for the NO+CO reaction on 1, 2, and 3 monolayer (ML) Pd films supported by MgO(100) is calculated using density functional theory. Thin Pd films are generally found to be more reactive than thick films, with a notable exception for nitrogen adsorption on 2 ML Pd/MgO(100). For this system an attractive through-the-metal adsorbate-oxide interaction of 0.5 eV is identified. Nitrogen adsorption is consequently estimated to provide a thermodynamic driving force for the reconstruction of MgO(100) supported 3 ML (or thicker) Pd clusters into thinner Pd clusters. PMID- 12097031 TI - Measurement of the B(0) lifetime with partially reconstructed B(0)-->D( )l(+)nu(l) decays. AB - The B(0) lifetime was measured with a sample of 23 million BB pairs collected by the BABAR detector at the PEP-II e(+)e(-) storage ring during 1999 and 2000. Events from the semileptonic decay B(0)-->D(*-)l(+)nu(l) have been selected with a partial reconstruction method in which only the charged lepton and the slow pi from the D*--->D(0)pi(-) decay are reconstructed. The result is tau(B(0)) = 1.529+/-0.012(stat)+/-0.029(syst) ps. PMID- 12097056 TI - "Intrinsic" acceptor ground state splitting in silicon: an isotopic effect. AB - One of the oldest open questions in semiconductor physics is the origin of the small splittings of the neutral acceptor ground state in silicon which lead to a distribution of doublet splittings rather than the fourfold-degenerate state of Gamma(8) symmetry expected in the absence of perturbations. Here we show that these acceptor ground state splittings are absent in the photoluminescence spectra of acceptor bound excitons in isotopically purified 28Si, demonstrating conclusively the surprising result that the splittings previously observed in natural Si result from the randomness of the Si isotopic composition. PMID- 12097057 TI - Gapless spin-1 neutral collective mode branch for graphite. AB - Using the standard tight binding model of 2D graphite with short range electron repulsion, we predict a gapless spin-1, neutral collective mode branch below the particle-hole continuum with energy vanishing linearly with momenta at the Gamma and K points in the Brillouin zone. This spin-1 mode has a wide energy dispersion, 0 to approximately 2 eV, and is not Landau damped. The "Dirac cone spectrum" of electrons at the chemical potential of graphite generates our collective mode, so we call this "spin-1 zero sound" of the "Dirac sea." Epithermal neutron scattering experiments and spin polarized electron energy loss spectroscopy can be used to confirm and study our collective mode. PMID- 12097058 TI - Spin-orbital pattern dependent polaron absorption in manganites. AB - We systematically investigated optical properties of Nd1-xSrxMnO3 single crystals ( x = 0.40, 0.50, 0.55, and 0.65). They are similar in their spin-orbital (SO) disordered states at room temperature. At low temperature, the crystals enter into various SO ordered states, i.e., F-, CE-, A-, and C-type orderings, and their mid-infrared absorptions become quite different. The remarkable variation can be explained by polaron dynamics which depend on the ordering patterns. This SO pattern dependent polaron model can also explain the pseudo CE-type ordering case, demonstrating that this scheme can explain the carrier dynamics in complex SO configurations. PMID- 12097059 TI - Weak-localization-like temperature-dependent conductivity of a dilute two dimensional hole gas in a parallel magnetic field. AB - We have studied the magnetotransport properties of a high mobility two dimensional hole gas (2DHG) in a 10 nm GaAs quantum well with densities in the range of (0.7-1.6) x 10(10) cm(-2) on the metallic side of the zero-field "metal insulator transition." In a parallel field well above B(c) that suppresses the metallic conductivity, the 2DHG exhibits a conductivity Delta(g)(T) approximately (1/pi) (e(2)/h)lnT reminiscent of weak localization for Fermi liquids. The experiments are consistent with the coexistence of two phases in our system: a metallic phase and a weakly insulating Fermi liquid phase. PMID- 12097060 TI - Random resonators and prelocalized modes in disordered dielectric films. AB - We calculate the areal density of disorder-induced resonators with a high quality factor, Q>>1, in a film with fluctuating refraction index. We demonstrate that, for a given kl>1, where k is the light wave vector and l is the transport mean free path, when on average the light propagation is diffusive, the likelihood for finding a random resonator increases dramatically with increasing the correlation radius of the disorder. Parameters of most probable resonators as functions of Q and kl are found. PMID- 12097061 TI - Magic numbers in metallo-inorganic clusters: chromium encapsulated in silicon cages. AB - A systematic theoretical study of the equilibrium geometries and total energies of Cr encapsulated in Si clusters reveals that Cr@Si(12) is more stable than its neighbors. The origin of this enhanced stability is consistent with the 18 electron sum rule commonly used in the synthesis of stable chemical complexes, and may provide a criterion for a systematic search of magic numbers in metalloinorganic clusters. The 6 mu(B) magnetic moment of the caged Cr atom, the largest among the 3d transition metal atoms, is completely quenched. This effect of caging on the properties of transition metal atoms may lead to the synthesis of novel cluster based materials. PMID- 12097062 TI - Mesoscopic rectifiers based on ballistic transport. AB - Recent experiments on symmetry-broken mesoscopic semiconductor structures have exhibited an amazing rectifying effect in the transverse current-voltage characteristics with promising prospects for future applications. We present a simple microscopic model, which takes into account the energy dependence of current-carrying modes and explains the rectifying effect by an interplay of fully quantized and quasiclassical transport channels in the system. It also suggests the design of a ballistic rectifier with an optimized rectifying signal and predicts voltage oscillations which may provide an experimental test for the mechanism considered here. PMID- 12097063 TI - Frictional drag between two dilute two-dimensional hole layers. AB - We report drag measurements on dilute double layer two-dimensional hole systems in the regime of r(s) = 19-39. We observed a strong enhancement of the drag over the simple Boltzmann calculations of Coulomb interaction, and deviations from the T2 dependence which cannot be explained by phonon-mediated, plasmon-enhanced, or disorder-related processes. We suggest that this deviation results from interaction effects in the dilute regime. PMID- 12097064 TI - Superconductivity and magnetic fluctuations in Cd(2))Re(2)O(7) via Cd nuclear magnetic resonance and re nuclear quadrupole resonance. AB - We report Cd nuclear magnetic resonance (NMR) and Re nuclear quadrupole resonance (NQR) studies on Cd(2)Re(2)O(7), the first superconductor among pyrochlore oxides (T(c) approximately 1 K). The Re NQR spectrum at zero magnetic field below 100 K rules out any magnetic or charge order. The spin-lattice relaxation rate below T(c) exhibits a pronounced coherence peak and follows the weak-coupling BCS theory with nearly isotropic energy gap. The results of Cd NMR point to a moderate ferromagnetic enhancement at high temperatures followed by a rapid decrease of the density of states below the structural transition temperature of 200 K. PMID- 12097065 TI - Spontaneous formation of a pi soliton in a superconducting wire with an odd number of electrons. AB - We consider a one-dimensional superconducting wire where the total number of electrons can be controlled in the Coulomb blockade regime. We predict that a pi soliton (kink) will spontaneously form in the system when the number of electrons is odd, because this configuration has a lower energy. If the wire with an odd number of electrons is closed in a ring, the phase difference on the two sides of the soliton will generate a supercurrent detectable by a SQUID. The two degenerate states with the current flowing clockwise or counterclockwise can be utilized as a qubit. PMID- 12097066 TI - Proximity of pseudogapped superconductor and commensurate antiferromagnet in a quasi-two-dimensional organic system. AB - We performed the single-crystal 13C NMR studies on a quasi-two-dimensional system, deuterated kappa-(BEDT-TTF)2Cu[N(CN)(2)]Br, which is just on the border of the Mott transition. The NMR spectra are separated into two parts coming from the metallic (superconducting) and insulating phases due to the phase separation at low temperature. The examination of the separated spectra revealed that the Mott transition in this system is characterized by the first-order transition between the pseudogapped superconductor and the simplest commensurate antiferromagnet with a moment of 0.26 mu(B)/dimer. PMID- 12097067 TI - London theory for superconducting phase transitions in external magnetic fields: application to UPt3. AB - For multicomponent superconductors, it is known that the presence of symmetry breaking fields can lead to multiple superconducting phase transitions. Motivated by recent small angle neutron scattering experiments on the vortex state of UPt3, the London theory in the vicinity of such phase transitions is determined. It is found that the form of this London theory is in general quite different than that for conventional superconductors. This is due to the existence of a diverging correlation length associated with these phase transitions. One striking consequence is that nontrivial vortex lattices exist arbitrarily close to H(c1). Applications to UPt3, CeIn3, U(1-x)Th(x)Be(13), electron doped cuprate superconductors, Sr(2)RuO(4), and MgCNi(3) are discussed. PMID- 12097068 TI - Tunneling between dissimilar high-T(c) oxide superconductors. AB - We report the first successful fabrication and measurement of high-T(c) heterojunctions with different oxide electrodes, YBa(2)Cu(3)O(7-y) and Bi(2)Sr(2)CaCu(2)O(y). Different kinds of junction characteristics are observable according to the magnitude of the tunnel resistance. With higher tunnel resistance, gap structures corresponding to two gaps are clearly observed, ensuring that the conventional tunneling scheme is also valid for this geometry. Peculiar behavior for the zero bias conductance peak is also observable. Josephson current is found to flow between these dissimilar superconductors. PMID- 12097069 TI - Magneto-optics in the ultrafast regime: thermalization of spin populations in ferromagnetic films. AB - We have characterized by pump-probe polarimetry the time-dependent dielectric tensor in a CoPt3 ferromagnetic film excited by 20 fs laser pulses. It is shown that, after the thermalization time of the electrons (approximately 50 fs), the dynamics of the real and the imaginary parts of the Voigt vector are identical. In addition, their relative variation is 10 times larger than that of the diagonal elements of the tensor, which allows one to infer that the spins dominate the magneto-optical response. During the thermalization process, the temporal behavior of the tensor elements opens new questions concerning the dynamics of the spins associated to a nonthermal electronic population in a ferromagnet. PMID- 12097070 TI - Large four-wave mixing of spatially extended excitonic states in thin GaAs layers. AB - We study the size dependence of the nonlinear response of weakly confined excitons for the size region beyond the long wavelength approximation regime. The observed degenerate-four-wave mixing signal of GaAs thin layers exhibits an anomalous size dependence, where the signal is resonantly enhanced at a particular thickness region. The theoretical analysis elucidates that this enhancement is due to the size-resonant enhancement of the internal field with a spatial structure relevant to the nondipole-type excitonic state. These results establish the formerly proposed new type of size dependence of nonlinear response due to the nonlocality induced double resonance. PMID- 12097071 TI - All-silicon quantum computer. AB - A solid-state implementation of a quantum computer composed entirely of silicon is proposed. Qubits are 29Si nuclear spins arranged as chains in a 28Si (spin-0) matrix with Larmor frequencies separated by a large magnetic field gradient. No impurity dopants or electrical contacts are needed. Initialization is accomplished by optical pumping, algorithmic cooling, and pseudo-pure state techniques. Magnetic resonance force microscopy is used for ensemble measurement. PMID- 12097072 TI - Model of inversion of DNA charge by a positive polymer: fractionalization of the polymer charge. AB - We model one strand of DNA by a one-dimensional lattice (ODL) of negative charges and consider the problem of inversion of its charge by a positive polyelectrolyte (PE). In the neutral state of the ODL-PE complex, each of the ODL charges is locally compensated by a PE charge. When an additional PE molecule is adsorbed by ODL, its charge gets fractionalized into monomer charges of defects (tails and arches) on the background of the perfectly neutralized ODL. Defects spread all over the ODL, eliminating the self-energy of PE. For DNA this fractionalization mechanism leads to a substantial inversion of charge, a phenomenon which is widely used for gene delivery. PMID- 12097073 TI - Quantum-classical transition induced by electrical measurement. AB - A model of an electrical tunnel junction coupled to a mechanical system (oscillator) is studied to simulate the dephasing effect of measurement on a quantum system. The problem is solved at zero temperature under conditions of strong nonequilibrium in the measurement apparatus. For linear coupling between the oscillator and tunneling electrons, it is found that the oscillator dynamics becomes damped, with the effective temperature determined by the voltage drop across the junction. It is demonstrated that both the quantum heating and the quantum damping of the oscillator manifest themselves in the current-voltage characteristic of the tunnel junction. PMID- 12097074 TI - Dynamics of folding in semiflexible filaments. AB - The dynamics of a single semiflexible filament under the action of a compressing force is simulated. We find that the filament folds asymmetrically with a folding length which depends only on the bending stiffness kappa and the applied force f. It is shown that this behavior can be attributed to the exponentially falling tension profile in the filament. While the folding time tau(0) depends on the initial configuration, the distance moved by the terminal point of the filament and the length of the fold scales as tau(1/2) at tau>>tau(0) and is independent of the initial configuration. PMID- 12097075 TI - Phase behavior of columnar DNA assemblies. AB - The interaction between two stiff parallel DNA molecules depends not only on the distance between their axes but also on their azimuthal orientation. The positional and orientational order in columnar B-DNA assemblies in solution is investigated, on the basis of the electrostatic pair potential that takes into account DNA helical symmetry and the amount and distribution of adsorbed counterions. A phase diagram obtained by lattice sums predicts a variety of positionally and azimuthally ordered phases and bundling transitions strongly depending on the counterion adsorption patterns. PMID- 12097076 TI - Quantum-optical catalysis: generating nonclassical states of light by means of linear optics. AB - We report preparation and characterization of coherent superposition states t[0>+alpha]1> of the electromagnetic field by conditional measurements on a beam splitter. This state is generated in one of the beam splitter output channels if a coherent state [alpha> and a single-photon Fock state [1> are present in the two input ports and a single photon is registered in the other beam splitter output. The single photon thus plays a role of a "catalyst:" it is explicitly present in both the input and the output channels of the interaction yet facilitates generation of a nonclassical state of light. PMID- 12097077 TI - Transverse breathing mode of an elongated Bose-Einstein condensate. AB - We study experimentally the transverse monopole mode of an elongated rubidium condensate. Because of the scaling invariance of the nonlinear Schrodinger (Gross Pitaevskii) equation, the oscillation is monochromatic and sinusoidal at short times, even under strong excitation. For ultralow temperatures, the quality factor Q = omega(0)/gamma(0) can exceed 2000, where omega(0) and gamma(0) are the mode angular frequency and damping rate. This value is much larger than any previously reported for other eigenmodes of a condensate. We also present the temperature variation of omega(0) and gamma(0). PMID- 12097078 TI - Critical rotation of a harmonically trapped Bose gas. AB - We study experimentally and theoretically a cold trapped Bose gas under critical rotation, i.e., with a rotation frequency close to the frequency of the radial confinement. We identify two regimes: the regime of explosion where the cloud expands to infinity in one direction, and the regime where the condensate spirals out of the trap as a rigid body. The former is realized for a dilute cloud, and the latter for a condensate with the interparticle interaction exceeding a critical value. This constitutes a novel system in which repulsive interactions help in maintaining particles together. PMID- 12097080 TI - Space-adiabatic perturbation theory in quantum dynamics. AB - A systematic perturbation scheme is developed for approximate solutions to the time-dependent Schrodinger equation with a space-adiabatic Hamiltonian. For a particular isolated energy band, the basic approach is to separate kinematics from dynamics. The kinematics is defined through a subspace of the full Hilbert space for which transitions to other band subspaces are suppressed to all orders, and the dynamics operates in that subspace in terms of an effective intraband Hamiltonian. As novel applications, we discuss the Born-Oppenheimer theory to second order and derive for the first time the nonperturbative definition of the g factor of the electron within nonrelativistic quantum electrodynamics. PMID- 12097079 TI - Specular reflection of matter waves from a rough mirror. AB - We present a high resolution study of the specularity of the atomic reflection from an evanescent wave mirror using velocity selective Raman transitions. We observed a double structure in the velocity distribution after reflection: a peak consistent with specular reflection and a diffuse reflection pedestal whose contribution decreases rapidly with increasing detuning. The diffuse reflection is due to two distinct effects: spontaneous emission in the evanescent wave and roughness in the evanescent wave potential whose amplitude is smaller than the de Broglie wavelength of the reflected atoms. PMID- 12097081 TI - Birefringence effect as a tool for astrophysical plasma study. AB - Group delay between two magnetoionic modes is discovered and spectrally confirmed ( deltat(g) proportional, variantf(-3)) for the first time for quasiperiodic narrow band millisecond solar radio pulsations. Analysis of cross delays and autodelays proves that both X and O modes originate at the same source, so the radiation is weakly polarized originally. This finding allows one to specify the emission mechanism of the pulsations to be a nonlinear plasma mechanism operating at the second harmonics. Physical parameters of the solar corona are determined by the use of the theory of the nonlinear plasma mechanism; they agree well with independent observations of solar millisecond pulsations. PMID- 12097082 TI - QCD-like behavior of high-temperature confining strings. AB - We show that, contrary to previous string models, the high-temperature behavior of the recently proposed confining strings reproduces exactly the correct large- N QCD result, a necessary condition for any string model of confinement. PMID- 12097083 TI - Superallowed beta decay of nuclei with a >or= 62: the limiting effect of weak Gamow-Teller branches. AB - The most precise value of V(ud), which is obtained from superallowed nuclear beta decay, leads to a violation of Cabibbo-Kobayashi-Maskawa unitarity by 2.2sigma. Experiments are underway on two continents to test and improve this result through decay studies of odd-odd N = Z nuclei with A>or=62. We show, in a series of illustrative shell-model calculations, that numerous weak Gamow-Teller branches are expected to compete with the superallowed branch in each of these nuclei. Though the total Gamow-Teller strength is significant, many of the individual branches will be unobservably weak. Thus, new techniques must be developed if reliable ft values are to be obtained with 0.1% precision for the superallowed branches. PMID- 12097084 TI - Nuclear tetrahedral symmetry: possibly present throughout the periodic table. AB - More than half a century after the fundamental, spherical shell structure in nuclei had been established, theoretical predictions indicated that the shell gaps comparable or even stronger than those at spherical shapes may exist. Group theoretical analysis supported by realistic mean-field calculations indicate that the corresponding nuclei are characterized by the TD(d) ("double-tetrahedral") symmetry group. Strong shell-gap structure is enhanced by the existence of the four-dimensional irreducible representations of TD(d); it can be seen as a geometrical effect that does not depend on a particular realization of the mean field. Possibilities of discovering the TD(d) symmetry in experiment are discussed. PMID- 12097085 TI - Spontaneous breaking of rotational symmetry in superconductors. AB - We show that homogeneous superconductors with broken spin/isospin symmetry lower their energy via a transition to a novel superconducting state where the Fermi surfaces are deformed to a quasiellipsoidal form at zero total momentum of Cooper pairs. In this state, the gain in the condensation energy of the pairs dominates over the loss in the kinetic energy caused by the lowest order (quadrupole) deformation of Fermi surfaces from the spherically symmetric form. The phase transition from the spherically symmetric state to the superconducting state with broken rotational symmetry is first order. PMID- 12097086 TI - Sympathetic cooling with two atomic species in an optical trap. AB - We simultaneously trap ultracold lithium and cesium atoms in an optical dipole trap formed by the focus of a CO2 laser and study the exchange of thermal energy between the gases. The optically cooled cesium gas efficiently decreases the temperature of the lithium gas through sympathetic cooling. Equilibrium temperatures down to 25 microK have been reached. The measured cross section for thermalizing 133Cs-7Li collisions is 8 x 10(-12) cm(2), for both species unpolarized in their lowest hyperfine ground state. Besides thermalization, we observe evaporation of lithium purely through elastic cesium-lithium collisions (sympathetic evaporation). PMID- 12097087 TI - Critical properties of the synchronization transition in space-time chaos. AB - We study two coupled spatially extended dynamical systems which exhibit space time chaos. The transition to the synchronized state is treated as a nonequilibrium phase transition, where the average synchronization error is the order parameter. The transition in one-dimensional systems is found to be generically in the universality class of the Kardar-Parisi-Zhang equation with a growth-limiting term ("bounded KPZ"). For systems with very strong nonlinearities in the local dynamics, however, the transition is found to be in the universality class of directed percolation. PMID- 12097088 TI - Experimental measurements of stretching fields in fluid mixing. AB - Using precision measurements of tracer particle trajectories in a two-dimensional fluid flow producing chaotic mixing, we directly measure the time-dependent stretching field. This quantity, previously available only numerically, attains local maxima along lines that coincide with the stable and unstable manifolds of hyperbolic fixed points of Poincare maps. Contours of a passive impurity field are found at each instant to be oriented parallel to the lines that have recently experienced large stretching. The local stretching varies by 12 orders of magnitude. PMID- 12097089 TI - Steady-state far-infrared coherent synchrotron radiation detected at BESSY II. AB - At BESSY II it is demonstrated that far-infrared coherent synchrotron radiation (CSR) can be generated by a controlled, steady-state process at storage rings. As an indication for coherent emission, the radiated power grows with the square of the beam current. The spectrum was analyzed by an interferometer in the 1-mm to 0.3-mm wavelength range. The CSR was enhanced more than 3000 times above background; the incoherent radiation remained below the background level. Steady state and bursting CSR were discriminated by time resolved analysis from micro seconds to seconds. PMID- 12097090 TI - Anisotropic free-energy limit of halos in high-intensity accelerators. AB - We study halo emittance growth in anisotropic beams and show that the rms emittance growth resulting from mismatch is highly anisotropic, depending on the tune ratio. We find that the free-energy limit calculated by Reiser [J. Appl. Phys. 70, 1919 (1991)] for an axisymmetric 1D halo can be extended to 2D if understood as an upper bound to the rms emittance growth averaged per degree of freedom. The thus-obtained "free-energy limit" of an ideal transport system is compared with the halo rms emittance growth in simulations of the Spallation Neutron Source linac. PMID- 12097091 TI - Fluctuation-dissipation relations for a nonlocal plasma. AB - A generalized version of the Callen-Welton fluctuation-dissipation formula that is nonlocal in space and time is derived. In a nonuniform plasma there appear significant differences between the fluctuations of the electrostatic field and those of the electron density, and the spatial inhomogeneities lead to an asymmetry with respect to the sign inversion of the frequency. PMID- 12097092 TI - Slow L-H transitions in DIII-D plasmas. AB - The transition from the low to the high mode of plasma confinement ( L-H transition) is studied in the DIII-D by an experimental technique which allows an arbitrarily slow transition. During an initial transition, periodic turbulent instability bursts are observed near the separatrix which inhibit the full transition. These bursts are damped by self-generated shear flows, and a predator prey-type relationship is shown to give a good description of the data. As the neutral-beam power is raised, the oscillations change to type III edge localized modes. Another transition then leads to a quiet H mode. PMID- 12097093 TI - Potential-energy landscapes of simple liquids. AB - Changes in the potential-energy surface as a function of the range and curvature of the pair potential are studied using isothermal-isobaric ensemble Monte Carlo simulations of Morse liquids. The configurational energies of stationary points are found to be linear functions of the fraction of imaginary modes, with slopes that are proportional to the range of the potential. The relative energies of saddles, minima, and instantaneous configurations show qualitatively different behavior for short, long, and intermediate range potentials, which imply corresponding variations in liquid state relaxation dynamics. PMID- 12097094 TI - Nanostructure evolution during cluster growth: Ag on H-terminated Si(111) surfaces. AB - The size of small clusters has a large effect on cluster growth and structure. Using Ag clusters of sizes from approximately 1 to 10 nm grown on H-Si(111), we found that the cluster structure evolves in stages showing (1) irregular shapes at coverage interstitial loops in ferritic materials. AB - We propose a comprehensive mechanism for the formation and growth of <100> interstitial loops in alpha-Fe. This mechanism reconciles long-standing experimental observations of these defects in irradiated ferritic materials with recent atomistic simulations of collision cascades and defect cluster properties in Fe, in which highly mobile 1 / 2<111> clusters are seen to be the dominant feature. Hence, this work provides one of the necessary links to unify simulation with experiments in alpha-Fe and ferritic alloys subject to high-energy particle irradiation. PMID- 12097100 TI - Pressure-induced structural change of liquid silicon. AB - High-pressure and high-temperature x-ray diffraction measurements indicate that liquid silicon contracts with increasing pressure without significant changes in the local structure up to 8 GPa and then transforms to a denser structure between 8 and 14 GPa. In spite of volume contraction, the nearest-neighbor interatomic distance expands by about 1.6% within this pressure interval, accompanied by an anomalous increase in the coordination number. These findings reveal that the drastic pressure-induced structural change can take place in three-dimensional network liquids with rather isotropic bonding. PMID- 12097101 TI - Tunneling state anisotropy in a single grain decagonal quasicrystal. AB - We present low-temperature measurements, between 20 mK and 20 K, of the sound velocity variations in a single grain of the decagonal quasicrystal Al-Ni-Co. We have used acoustic shear waves of frequency 70 MHz propagating in the quasicrystalline planes with two different polarizations: parallel and perpendicular to the quasicrystalline planes. Our results show the presence of tunneling states (TSs) with a strong difference of the phonon-TS coupling for the two polarizations, which reveals a strong anisotropy of distribution of the orientations of the TSs in the sample. This result gives direct evidence that TSs are generated by the quasicrystalline structure itself. PMID- 12097102 TI - Atomic structures of the Ge/Si(113)-(2 x 2) surface. AB - Based on scanning tunneling microscopy observations of the epitaxial growth of Ge on Si(113) and first-principles total energy and band calculations, we demonstrate that the Ge/Si(113)-(2 x 2) surface is made up of alternating [1;10] oriented rows of rebonded atoms and tilted pentamers of five atoms, where each pentamer is stabilized by an interstitial atom at the subsurface. From the existence of stacking defects in rows of tilted pentamers observed at room temperature, we have deduced that at epitaxial temperatures the pentamers frequently change their tilting orientations between two minimum energy states. PMID- 12097103 TI - Adsorption in noninterconnected pores open at one or at both ends: a reconsideration of the origin of the hysteresis phenomenon. AB - We report on an experimental study of adsorption isotherm of nitrogen onto porous silicon with noninterconnected pores open at one or at both ends in order to check for the first time the old (1938) but always current idea based on Cohan's description which suggests that the adsorption of gas should occur reversibly in the first case and irreversibly in the second one. Hysteresis loops, the shape of which is usually associated with interconnections in porous media, are observed whether the pores are open at one or at both ends, in contradiction with Cohan's model. PMID- 12097104 TI - Critical role of the surface reconstruction in the thermodynamic stability of (105) Ge pyramids on Si(001). AB - We show by molecular dynamics simulations on a scale comparable to experimental dimensions that a peculiar surface reconstruction of the (105) facets is responsible for the stability of Ge pyramids on Si(001). This finding is confirmed by ab initio total energy calculations for competing surface reconstructions and a very satisfactory comparison of the corresponding charge density maps to scanning tunneling microscopy measurements of the facets, both for filled and empty states. PMID- 12097105 TI - Influence of external stress on surface reaction dynamics. AB - In exploring the dynamics of dissociative chemisorption of O2 on clean and O covered Cu(001), we demonstrate that the chemisorption is very sensitive to uniaxial tensile stress within an elastic limit. The stress enhances the dissociative chemisorption on clean Cu(001) when the translational energy of incident O2 is below 250 meV, and suppresses it when the energy is above 250 meV. In the case of an oxygen-covered Cu(001)- (2squareroot2 x squareroot2) surface, the dissociative adsorption probability of O2 decreases as the stress increases. The effect of external stress on the O2 dissociation dynamics is different between clean and O-covered surfaces. PMID- 12097106 TI - One-dimensional disordered density waves and superfluids: the role of quantum phase slips and thermal fluctuations. AB - The low temperature phase diagram of 1D disordered quantum systems such as charge or spin density waves, superfluids, and related systems is considered by a full finite- T renormalization group approach for the first time. At zero temperature the consideration of quantum phase slips leads to a new scenario for the unpinning (delocalization) transition. In the strong pinning limit the model is solved exactly. At finite T a rich crossover diagram with various scaling regions is found which reflects the zero temperature quantum critical behavior. PMID- 12097107 TI - Charge-density patching method for unconventional semiconductor binary systems. AB - A motif based charge patching method is presented for large system electronic structure calculations. It produces ab initio quality charge densities for large systems without actually doing self-consistent calculations for them. It represents a general faster alternative to the conventional O(N) methods. This method is applied here to unconventional semiconductor binary systems, and the resulting eigenenergies are found to be almost the same as the original ab initio eigenenergies (with 20-50 meV errors). PMID- 12097108 TI - Time-dependent density-matrix renormalization group: a systematic method for the study of quantum many-body out-of-equilibrium systems. AB - The density-matrix renormalization-group algorithm is extended to treat time dependent problems. The method provides a systematic and robust tool to explore out-of-equilibrium phenomena in quantum many-body systems. We illustrate the method by showing that attractive interactions enhance the tunneling current between two Luttinger liquids, whereas repulsive interactions suppress it, in qualitative agreement with analytical predictions. Enhancement of the transport current through a quantum dot in the Kondo regime is also exhibited. PMID- 12097109 TI - Correlation energy and spin polarization in the 2D electron gas. AB - The ground-state energy of the two-dimensional uniform electron gas has been calculated with a fixed-node diffusion Monte Carlo method, including backflow correlations, for a wide range of electron densities as a function of spin polarization. We give a simple analytic representation of the correlation energy which fits our simulation data and includes several known high- and low-density limits. This parametrization provides a reliable local spin density energy functional for two-dimensional systems and an estimate for the spin susceptibility. Within the proposed model for the correlation energy, a weakly first-order polarization transition occurs shortly before Wigner crystallization as the density is lowered. PMID- 12097110 TI - Manipulation of the spin memory of electrons in n-GaAs. AB - We report on the optical manipulation of the electron spin relaxation time in a GaAs-based heterostructure. Experimental and theoretical study shows that the average electron spin relaxes through hyperfine interaction with the lattice nuclei, and that the rate can be controlled by electron-electron interactions. This time has been changed from 300 ns down to 5 ns by variation of the laser frequency. This modification originates in the optically induced depletion of an n-GaAs layer. PMID- 12097111 TI - Large electron-phonon coupling at an interface. AB - The strength of electron-phonon coupling in atomically uniform films of Ag on Fe is determined by angle-resolved photoemission from quantum well states in these films over a wide temperature range. As the film thickness is reduced, contributions from the surface and interface should become more important, and, experimentally, a large enhancement with superimposed quantum oscillations is observed. An analysis of the quantum oscillations indicates that this large enhancement is an interface effect. PMID- 12097112 TI - Electrical transport through single-molecule junctions: from molecular orbitals to conduction channels. AB - We present an atomistic theory of electronic transport through single organic molecules that reproduces the important features of the current-voltage ( I-V) characteristics observed in recent experiments. We trace these features to their origin in the electronic structure of the molecules and their local atomic environment. We demonstrate how conduction channels arise from the molecular orbitals and elucidate the contributions of individual orbitals to the current. We find that in thiol-bridged aromatic molecules many molecular orbitals contribute to a single conduction channel and discuss the implications of this result for the design of molecular devices. PMID- 12097113 TI - Collapse of the spin-singlet phase in quantum dots. AB - We present experimental and theoretical results on a new regime in quantum dots in which the filling factor two-singlet state is replaced by new spin polarized phases. We make use of spin blockade spectroscopy to identify the transition to this new regime as a function of the number of electrons. The key experimental observation is a reversal of the phase in the systematic oscillation of the amplitude of Coulomb blockade peaks as the number of electrons is increased above a critical number. It is found theoretically that correlations are crucial to the existence of the new phases. PMID- 12097114 TI - Conductance fluctuations and weak localization in chaotic quantum dots. AB - We study the conductance statistical features of ballistic electrons flowing through a chaotic quantum dot. We show how the temperature affects the universal conductance fluctuations by analyzing the influence of dephasing and thermal smearing. This leads us to two main findings. First, we show that the energy correlations in the transmission, which were overlooked so far, are important for calculating the variance and higher moments of the conductance. Second, we show that there is an ambiguity in the method of determination of the dephasing rate from the size of the weak localization. We find that the dephasing times obtained at low temperatures from quantum dots are underestimated. PMID- 12097115 TI - Tuning of the Fano effect through a quantum dot in an Aharonov-Bohm interferometer. AB - The Fano effect, which arises from an interference between a localized state and the continuum, reveals a fundamental aspect of quantum mechanics. We have realized a tunable Fano system in a quantum dot (QD) in an Aharonov-Bohm interferometer, which is the first convincing demonstration of this effect in mesoscopic systems. With the aid of the continuum, the localized state inside the QD acquires itinerancy over the system even in the Coulomb blockade. Through tuning of the parameters, which is an advantage of the present system, unique properties of the Fano effect on the phase and coherence of electrons have been revealed. PMID- 12097117 TI - Parametric spectral correlations in disordered and chaotic structures. AB - We explore the influence of external perturbations on the energy levels of a Hamiltonian drawn at random from the Gaussian unitary distribution of Hermitian matrices. By deriving the joint distribution function of eigenvalues, we obtain the (n,m)-point parametric correlation function of the initial and the final density of states for perturbations of arbitrary rank and strength. A further generalization of these results allows for the incorporation of short-range spatial correlations in diffusive as well as ballistic chaotic structures. PMID- 12097116 TI - Resistively detected nuclear magnetic resonance in the quantum hall regime: possible evidence for a Skyrme crystal. AB - Resistively detected nuclear magnetic resonance measurements have been performed on a high mobility heterostructure in the quantum Hall regime. At millikelvin temperatures the nuclear resonances are observed in the vicinity of various integer and fractional filling factors without previous dynamic nuclear polarization. Near nu = 1, the observed large enhancement of the resonance amplitude accompanied by a reduction of T1 strongly suggests a greatly increased coupling between the electronic and nuclear spin systems. This is consistent with the proposed coupling of the nuclear spin system to the Goldstone mode of the Skyrme crystal. PMID- 12097118 TI - Doping dependence of an n-type cuprate superconductor investigated by angle resolved photoemission spectroscopy. AB - We present an angle-resolved photoemission doping dependence study of the n-type cuprate superconductor Nd(2-x)Ce(x)CuO(4+/-delta), from the half-filled Mott insulator to the T(c) = 24 K superconductor. In Nd2CuO4, we reveal the charge transfer band for the first time. As electrons are doped into the system, this feature's intensity decreases with the concomitant formation of near- E(F) spectral weight. At low doping, the Fermi surface is an electron-pocket (with volume approximately x) centered at (pi,0). Further doping leads to the creation of a new holelike Fermi surface (volume approximately 1+x) centered at (pi,pi). These findings shed light on the Mott gap, its doping evolution, as well as the anomalous transport properties of the n-type cuprates. PMID- 12097119 TI - Spin-1 neutron resonance peak cannot account for electronic anomalies in the cuprate superconductors. AB - In certain cuprates, a spin-1 resonance mode is prominent in the magnetic structure measured by neutron scattering. It has been proposed that this mode is responsible for significant features seen in other spectroscopies, such as photoemission and optical absorption, which are sensitive to the charge dynamics, and even that this mode is the boson responsible for "mediating" the superconducting pairing. We show that its small (measured) intensity and weak coupling to electron-hole pairs (as deduced from the measured lifetime) disqualifies the resonant mode from either proposed role. PMID- 12097121 TI - Nonlinear ac resistivity in s-wave and d-wave disordered granular superconductors. AB - We model s-wave and d-wave disordered granular superconductors with a three dimensional lattice of randomly distributed Josephson junctions. The nonlinear ac resistivity rho(2) of these systems was calculated using Langevin dynamical equations. The current amplitude dependence of rho(2) at the peak position is found to be a power law characterized by exponent alpha, which is not universal but depends on the self-inductance and current regimes. In the weak current regime alpha is independent of the self-inductance and alpha = 0.5+/-0.1 for both s- and d-wave materials. In accord with experiments, we find alpha approximately 1 for some interval of inductance in the strong current regime. PMID- 12097120 TI - High field phase diagram of cuprates derived from the Nernst effect. AB - Measurements of the Nernst signal in the vortex-liquid state of the cuprates to high fields (33 T) reveal that vorticity extends to very high fields even close to the zero-field critical temperature T(c0). In overdoped La2-xSrxCuO4, we show that the upper critical field H(c2)(T) curve does not end at T(c0), but at a much higher temperature. These results imply that T(c0) corresponds to a loss in phase rigidity rather than a vanishing of the pairing amplitude. An intermediate field H*(T)<